PMID- 17907823 TI - Dopamine D1 receptor stimulation of the nucleus accumbens or the medial preoptic area promotes the onset of maternal behavior in pregnancy-terminated rats. AB - There is good evidence that interference with the mesolimbic dopamine (DA) system results in impaired maternal responding in postpartum female rats. However, whether activation of the mesolimbic DA system is capable of promoting maternal behavior has not been investigated. This study examined whether increasing DA activity in various brain regions of pregnancy-terminated, naive female rats would stimulate the onset of maternal behavior. Experiments 1 and 2 examined the effects of microinjection of various doses (0, 0.2, or 0.5 microg/0.5 microl/side) of a D1 DA receptor agonist, SKF 38393, or a D2 DA receptor agonist, quinpirole, into the nucleus accumbens (NA) on latency to show full maternal behavior, and Experiment 3 determined the effects of SKF 38393 injection into a control site. Finally, because the medial preoptic area (MPOA) is also important for maternal behavior, receives DA input, and expresses DA receptors, the authors examined whether microinjection of SKF 38393 into MPOA was capable of stimulating the onset of maternal behavior. Results indicated that microinjection of SKF 38393 into either the NA or the MPOA facilitates maternal responding in pregnancy terminated rats. PMID- 17907822 TI - Differential involvement of the basolateral amygdala, orbitofrontal cortex, and nucleus accumbens core in the acquisition and use of reward expectancies. AB - In this study, the authors tested the hypothesis that the basolateral amygdala (BLA), orbitofrontal cortex (OFC), nucleus accumbens core (NA-core), and the extended hippocampus mediate different aspects of the development-maintenance of unique reward expectancies produced by the differential outcomes procedure (DOP). Rats were trained with either DOP or a nondifferential outcomes procedure (NOP) on a simple discrimination task. Fornix lesions did not affect either version of the task, demonstrating that the extended hippocampal system has no role in stimulus-outcome (S-O) associations. In contrast, in the DOP condition, BLA lesions impaired performance throughout training, OFC lesions impaired choice accuracy only in the later maintenance phase, and NA-core lesions resulted in enhanced learning. These results suggest that BLA and OFC are important for establishment (BLA) and behavioral maintenance (OFC) of S-O associations, whereas the NA-core is not needed and can in fact impede using multiple S-O associations. No impairments were observed in the NOP condition, demonstrating that these structures are not critical to stimulus-response learning. PMID- 17907824 TI - Effects of dopaminergic drugs on innate pheromone-mediated reward in female mice: a new case of dopamine-independent "liking.". AB - Male sexual pheromones are innately rewarding to adult female mice, but the role of dopamine in this natural reward is unknown. The authors have tackled this issue by assessing the effects of intraperitoneal injections of dopamine D1 (SCH 23390, 0.02- 0.05 mg/kg) and D2 (sulpiride, 20.00 mg/kg) antagonists, a dopamine releasing agent (amphetamine, 0.50 -2.00 mg/kg), and D1 (SKF 38393, 10.00 -20.00 mg/kg) and D2 (quinpirole, 0.20 -1.00 mg/kg) agonists on the chemoinvestigation displayed by female mice in male- versus female-soiled bedding 2-choice tests. Dopamine antagonists and quinpirole failed to affect the unconditioned preference displayed by females towards male chemosignals, whereas both amphetamine and SKF 38393 abolished it. Finally, D1 and D2 antagonists did not block the induction of operant place conditioning by male chemosignals. As the female mice were tested in their first encounter with male sexual pheromones, their behavior can only be influenced by the "liking" component of reward. Therefore, the results suggest that dopamine mediates neither the hedonic properties of male sexual pheromones nor the acquisition of conditioned place preference. However, dopamine acting on D1 receptors might inhibit female mice attraction towards male chemosignals. PMID- 17907825 TI - Roles of central catecholamine and hypothalamic neuropeptide Y genome in the development of tolerance to phenylpropanolamine-mediated appetite suppression. AB - Phenylpropanolamine (PPA) is an appetite suppressant. Repeated treatment with PPA can decrease food intake on initial days, but on subsequent days, food intake gradually returns to normal (tolerant effect). In an attempt to investigate the underlying mechanisms of PPA tolerance, the authors examined the roles of catecholamine (CAT) and hypothalamic neuropeptide Y (NPY) genome. Results revealed that pretreatment with either bupropion, a CAT transporter inhibitor, or a-methylparatyrosine, a tyrosine hydroxylase inhibitor, modulated the effect of PPA tolerance. Moreover, results also revealed that the alteration in NPY messenger RNA level coincided with the change of feeding behavior during PPA treatment and that infusions of NPY antisense oligonucleotide into the cerebroventricle abolished the effect of PPA tolerance. These findings suggest that cerebral CAT and hypothalamic NPY genome are involved in the development of tolerance to PPA-induced appetite suppression. PMID- 17907826 TI - Alterations in anterior hypothalamic vasopressin, but not serotonin, correlate with the temporal onset of aggressive behavior during adolescent anabolic androgenic steroid exposure in hamsters (Mesocricetus auratus). AB - In hamsters (Mesocricetus auratus), anabolic-androgenic steroid (AAS) exposure during adolescence facilitates offensive aggression that is correlated with the enhanced development of the arginine vasopressin (AVP) neural system and reduced development of the serotonin (5-HT) neural system in the anterior hypothalamus (AH). This study examined the temporal onset of these effects by measuring aggression and AH AVP and 5-HT during progressively shorter periods of AAS exposure during adolescent development. The authors tested adolescent hamsters that received AAS for 3, 7, 14, or 28 days for offensive aggression and then examined the hamsters for AVP/5-HT afferent innervation to the AH using immunohistochemistry. While reductions in AH 5-HT afferent innervation were detectable by 7 days of AAS exposure, no concomitant increases in offensive aggression were observed compared to oil-treated littermates. In contrast, by Day 14 of AAS treatment, AH AVP and offensive aggression were significantly higher than oil-treated controls. These data indicate that relatively short-term adolescent AAS exposure alters aggression and AH 5-HT and AVP development, yet only alterations in AH AVP development correlate with temporal onset of the aggressive behavioral phenotype during adolescent AAS exposure. PMID- 17907828 TI - Diet selection improves morphine's antinociceptive actions in rats with streptozotocin-induced diabetes. AB - Following the administration of the diabetogenic drug streptozotocin, rats selecting their diet from separate sources of macronutrients (e.g., proteins, fats, and carbohydrates) demonstrated less severe symptoms of diabetes than did rats fed ground Purina chow or a composite diet containing the same nutrient sources as found in the self-selection diet but in the proportions found in chow. After the induction of diabetes, rats selecting their own diet ate and drank less, weighed more, and had more adipose tissue and lower blood glucose levels than did rats consuming chow or the composite diet. In addition, rats choosing their diet were more sensitive to morphine's pain-relieving properties than were rats in the other 2 dietary groups. Rats given the self-selecting diet consumed more protein and fat and less carbohydrate than did those eating a single diet. Data suggest that rats must select their diet preceding and following the induction of diabetes for amelioration of diabetic symptoms to occur. These results indicate that diet can contribute to the severity of diabetes and could be used as an adjunct to standard treatment of the disease. PMID- 17907827 TI - Pharmacological modulation of stress reactivity dissociates general learning ability from the propensity for exploration. AB - It has previously been reported that general learning ability (GLA) correlates positively with exploratory tendencies in individual outbred mice. This finding suggests the possibility that variations in stress reactivity modulate GLA and thus its relationship to exploratory tendencies. Here, the authors investigated the potential role of stress reactivity in regulating this relationship by assessing the effects of the anxiolytic chlorodiazepoxide (CDP; 10 mg/kg) on subjects' performance in a battery of diverse learning tasks as well as exploratory behaviors and stress reactivity. CDP-treated mice exhibited reductions in stress-induced corticosterone levels and behavioral reactivity to mild stressors and a corresponding increase in exploration. However, CDP-treated mice did not exhibit facilitated acquisition of any of the learning tasks and expressed GLA comparable to controls. Results indicate that although reduced stress reactivity promotes exploration, this does not translate into an up regulation of GLA, suggesting that the relationship between GLA and exploration is not mediated by stress reactivity. The authors propose that variations in GLA reflect individuals' propensity for novelty seeking, whereas exploration reflects both stress reactivity and novelty seeking, the latter of which may underlie the relationship between exploration and GLA. PMID- 17907829 TI - Kappa-opioid receptor stimulation quickens pathogenesis of compulsive checking in the quinpirole sensitization model of obsessive-compulsive disorder (OCD). AB - Repeated injections of the D2/D3 dopamine agonist, quinpirole, induce locomotor sensitization and compulsive checking behavior in rats, a phenomenon that may constitute an animal model of obsessive- compulsive disorder (OCD). Considering that the co-joint treatment with quinpirole and the kappa opioid receptor agonist U69593 potentiates locomotor sensitization to quinpirole, the present study examined whether such co-stimulation of kappa and dopamine receptors also enhances compulsive checking and whether dopamine receptor supersensitivity mediates the augmentation effects. Results showed that co-treatment of quinpirole and U69593 had a robust accelerating effect on the acquisition of sensitized locomotion and compulsive checking but that the effects on the expression of quinpirole sensitization were behavior dependent, with increased magnitude of locomotion but not of compulsive checking. Quinpirole and even U69593, which by itself did not induce sensitization, increased the proportion of dopamine D2 receptors in the high-affinity state (D2(High)) in the nucleus accumbens and striatum, indicating that elevation of D2(High) is not sufficient to account for sensitization or compulsive checking. The animal model findings point to a potential role of kappa opioid systems in hastening the pathogenesis of OCD and to the possibility that distinct brain regions may mediate the development and the expression of compulsive checking. PMID- 17907830 TI - Therapeutic effects of exercise: wheel running reverses stress-induced interference with shuttle box escape. AB - Exercise can reduce symptoms of depression and anxiety in humans, but therapeutic effects of exercise in an animal model of stress-related mood disorders have yet to be demonstrated. In the current study, the authors investigated the ability of wheel running to reverse a long-lasting interference with shuttle box escape produced by uncontrollable stress. Rats who remained sedentary following uncontrollable foot shock demonstrated robust conditioned freezing behavior to the stressor environment and deficits in shuttle box escape learning. Voluntary access to running wheels for 6 weeks, but not 2 weeks, following uncontrollable foot shock reduced the expression of conditioned freezing and reversed the escape deficit. Results demonstrate a long-lasting interference with shuttle box escape that can be reversed by exercise in a duration-dependent fashion. PMID- 17907831 TI - Experience-dependent cell survival in the maternal rat brain. AB - Postpartum maternal experience produces long-lasting changes in maternal behavior in the mother rat, which can be altered by early-life isolation. Postpartum experience also affects the regulation of adult neurogenesis in the neural circuit underlying maternal behavior, in a region-specific manner. Female rats were reared either with their mothers (MR) or in isolation in an artificial rearing (AR) paradigm. In adulthood, rats were mated and separated from their pups at birth. The following day, dams were injected with a mitotic marker and either allowed to interact with pups (maternal experience) or left alone. Results show that MR rats that acquire a later maternal experience show increases in cell survival in parts of the excitatory limb of the maternal neural network (bed nucleus of the stria terminalis and nucleus accumbens), but no changes in the inhibitory limb (amygdala). In comparison to AR inexperienced rats, AR maternally experienced rats show no increases in cell survival in the excitatory limb, but a striking reduction in cell survival in the inhibitory limb. The results suggest that early preweaning maternal isolation alters the structural plasticity that occurs following a postpartum maternal experience. PMID- 17907832 TI - Activation of POMC neurons during general arousal but not sexual behavior in male rats. AB - Exogenous opioids influence male rat sexual behavior, suggesting that endogenous opioid peptides are released during mating. Supporting this hypothesis, the authors recently showed that mating induced activation of mu opioid receptors. However, it is unknown which ligand(s) is acting on these receptors during mating. The current set of experiments tested the hypothesis that beta-endorphin producing neurons, that is, proopiomelanocortin (POMC) neurons, are activated during sexual behavior. Mating-induced activation of POMC neurons was investigated during either the dark phase or the light phase, following different components of male rat sexual behavior or following control manipulations that resulted in general arousal. Results show activation of POMC neurons in the mediobasal hypothalamus following general arousal but not specifically related to sexual behavior per se. In addition, mating did not activate the subpopulation of POMC neurons that project to the medial preoptic nucleus. These results suggest that it is unlikely that POMC neurons contribute to the action of endogenous opioids in the brain area during sexual behavior but instead may contribute to the change in arousal state essential for the expression of sexual behavior. PMID- 17907833 TI - Mating activates NMDA receptors in the medial preoptic area of male rats. AB - Studies have emphasized the role of the medial preoptic area (MPOA) as an important site for the regulation of male sexual behavior. Indeed, ablations of the MPOA impair sexual behavior, whereas stimulation of the MPOA enhances behavior. Furthermore, neural activity in the MPOA increases with mating. The current study tested the hypothesis that activation of N-methyl-D-aspartate (NMDA) receptors occurs in MPOA neurons and is essential for the expression of male sexual behavior in rats. Results indicate that nearly all MPOA neurons that expressed Fos following mating also contained the NR1 subunit of NMDA receptors. Furthermore, mating increased phosphorylation, thus activation, of NR1 in the MPOA. Additionally, blocking NMDA receptors significantly decreased mating induced Fos expression and mating-induced phosphorylation of NMDA receptors and impaired male sexual behavior. These results provide evidence that mating activates NMDA receptors in the MPOA and that this activation is important for the expression of male sexual behavior. PMID- 17907834 TI - Rats spontaneously discriminate purely visual, two-dimensional stimuli in tests of recognition memory and perceptual oddity. AB - Animal models have been central to advances made in understanding the neural basis of human cognition, but maximizing the use of animal models requires tasks that match those used to assess human subjects. Tasks used in humans frequently use visual 2-dimensional stimuli, assess 1-trial learning, and require little pretraining. This article describes novel versions of 2 tasks for the rat, spontaneous object recognition and spontaneous oddity preference, both of which use purely visual, 2-dimensional picture-card stimuli, test 1-trial learning, and require no pretraining. Rats showed robust memory for a variety of picture-card stimuli, demonstrating almost no loss of memory for some of the stimulus types even after a 2-hr delay period. Rats were able to show spontaneous oddity preference for all 3 visual stimulus types tested (photos, shapes, and patterns), as well as for 3-dimensional objects. These 2 tasks are quick to administer, involve no fearful learning associations, and require a simple apparatus. They may be particularly useful for high-throughput pharmacological or genetic screening using rodent models. PMID- 17907835 TI - Temporal-difference prediction errors and Pavlovian fear conditioning: role of NMDA and opioid receptors. AB - Three experiments studied temporal-difference (TD) prediction errors during Pavlovian fear conditioning. In Stage I, rats received conditioned stimulus A (CSA) paired with shock. In Stage II, they received pairings of CSA and CSB with shock that blocked learning to CSB. In Stage III, a serial overlapping compound, CSB --> CSA, was followed by shock. The change in intratrial durations supported fear learning to CSB but reduced fear of CSA, revealing the operation of TD prediction errors. N-methyl- D-aspartate (NMDA) receptor antagonism prior to Stage III prevented learning, whereas opioid receptor antagonism selectively affected predictive learning. These findings support a role for TD prediction errors in fear conditioning. They suggest that NMDA receptors contribute to fear learning by acting on the product of predictive error, whereas opioid receptors contribute to predictive error. PMID- 17907836 TI - The ontogeny of fear-potentiated startle: effects of earlier-acquired fear memories. AB - Research has shown that learned fear emerges in a response-specific sequence. For example, freezing is observed at a younger age than is potentiated startle (P. Hunt & B. A. Campbell, 1997). The present study shows that the age at which a specific learned fear response emerges is influenced by the animal's early experiences. Specifically, fear potentiation of startle emerges earlier in development if the rat is given prior fear conditioning to a different stimulus. Some constraints of this "facilitation" effect are determined in follow-up experiments. This facilitation effect may provide a novel way of testing the development of the neural circuits underlying learned fear. PMID- 17907837 TI - Comparative behavioral effects between synthetic 2,4,5-trimethylthiazoline (TMT) and the odor of natural fox (Vulpes vulpes) feces in mice. AB - Synthetic 2,4,5-trimethylthiazoline (TMT)--a component of red fox (Vulpes vulpes) feces--is frequently used to induce unconditioned fear in rodents. Surprisingly, direct comparison between TMT and natural fox feces odor is almost nonexistent. In this study, Experiment 1 compared the avoidance in relation to TMT concentration, natural fox feces, and gender of fox and mice. Results show that the avoidance is (a) higher with either pure or 50% TMT as compared to natural fox feces, whereas the difference is slight with 10% TMT, and (b) significantly higher for the female mouse group compared to the male mouse group with TMT as well as natural fox feces. In addition, no clear difference in effect was observed between male and female fox feces. Experiment 2 compared behavioral parameters recorded as an index of fear and anxiety, general activity, and avoidance in elevated plus-maze and open-field chamber between 10% TMT and natural fox feces in relation to the estrus cycle of the mice. Results show no cycle period effect--except for the avoidance parameter "distance to odorant"- and no different effects between 10% TMT and natural fox feces except for freezing. PMID- 17907838 TI - Local anesthetic treatment significantly attenuates acute pain responding but does not prevent the neonatal injury-induced reduction in adult spinal behavioral plasticity. AB - Recent findings indicate that neonatal injury results in decreased spinal plasticity in adult subjects (E. E. Young, K. M. Baumbauer, A. E. Elliot, & R. L. Joynes, 2007). Previous research has shown that acute manipulations of pain processing (i.e., administration of formalin, carrageenan, capsaicin) result in a loss of spinal behavioral plasticity (A. R. Ferguson, E. D. Crown, & J. W. Grau, 2006). Moreover, neonatal injury results in a lasting reduction in adult spinally mediated plasticity resembling the deficit seen following acute manipulations in adults (E. E. Young et al., 2007). The present study was designed to determine whether the effects of neonatal injury could be prevented by lidocaine administration during the initial healing period. Subjects (injured or uninjured) received lidocaine or saline on 1 of 4 administration schedules (preinjury only, postinjury only, for 24 hr postsurgery, or for 72 hr postsurgery). Results demonstrated that lidocaine administration did not prevent the hypersensitivity and reduced spinal plasticity associated with neonatal injury. This suggests that (a) the mechanisms underlying neonatal injury are independent of peripheral input in the initial healing period and (b) lidocaine is ineffective at preventing long term spinal plasticity changes following neonatal injury. PMID- 17907839 TI - Neurokinin receptors modulate the impact of uncontrollable stimulation on adaptive spinal plasticity. AB - Previous research has demonstrated that spinally transected rats can acquire a prolonged flexion response to prevent the delivery of shock. However, rats that receive shock irrespective of leg position cannot learn to maintain the same response. The present experiments examined the role of neurokinin receptors in this learning deficit. Results demonstrated that neurokinin (NK1 and NK2) antagonists blocked the induction of the learning deficit, whereas NK agonists induced a learning deficit. The study found that NK agonist administration did not substitute for uncontrollable shock exposure. Finally, administration of an NK1 agonist prior to uncontrollable shock prevented the induction of the deficit. These results provide additional evidence that engaging nociceptive plasticity undermines the capability of spinal neurons to support adaptive changes. PMID- 17907840 TI - Neonatal hind-paw injury disrupts acquisition of an instrumental response in adult spinal rats. AB - The present study was designed to evaluate the impact of neonatal injury on adult spinal plasticity in rats. Subjects were randomly assigned to 1 of 4 experimental conditions: (a) hind-paw injury at Postnatal Day (PD) 2, (b) hind-paw injury at PD 5, (c) anesthesia exposure only on PD 2, or (d) anesthesia exposure only on PD 5. Subjects receiving a unilateral neonatal hind-paw injury showed decreased mechanical threshold (hyperalgesia) on the previously injured hind paw throughout development. This decrease in threshold survived spinal transection (at T2) at 12 weeks of age. Injured subjects also showed significant impairment in a spinal instrumental learning task performed by the previously injured hind paw. This disruption of learning indicates a disruption of spinal plasticity that may be due to induction of long-term changes in nociceptive processing within the spinal cord. PMID- 17907841 TI - Altered performance of reelin-receptor ApoER2 deficient mice on spatial tasks using the Barnes maze. AB - The apolipoprotein E receptor 2 (ApoER2), expressed predominantly in forebrain regions including the hippocampus, is 1 of 2 receptors for the extracellular matrix protein reelin, which is critical for cortical development. Previous studies of ApoER2 mutant mice have indicated deficits in synaptic plasticity and learning. The current authors assessed learning and memory of ApoER2 knockout and wild-type mice on the Barnes circular maze. Mice were trained in this task for 22 days, followed by memory recall and reversal tests. ApoER2 knockout mice were initially slower to complete the task, but by Day 22 they were more accurate than wild-type mice on several indices. There were no differences in memory assessed by the recall task, but ApoER2 knockout mice performed significantly worse on the memory reversal task. ApoER2 knockout mice also displayed altered use of specific search strategies and relationship of these strategies to errors made on the maze. PMID- 17907842 TI - Extracellular amino acid levels in the interpositus nucleus during classical eyeblink conditioning in alert cats. AB - The extracellular levels of selected amino acids in the cerebellar posterior interpositus nucleus (PIN) during classical eyeblink conditioning was analyzed in alert cats using a delay paradigm. Animals were prepared for the chronic recording of eyelid movements (with the magnetic search-coil technique) and the electromyographic activity of the orbicularis oculi muscle. With the help of a guide and push-pull cannulae, selected PIN sites were perfused daily during classical eyeblink conditioning. The perfusate was sampled at intervals of 5 min and analyzed with a high-pressure liquid chromatography- electrochemical detection (HPLC-EC) method. The analysis of push-pull perfusate revealed a significant increase in the release of glycine, taurine, and glutamate across the successive conditioning sessions, in parallel with the acquisition of eyelid conditioned responses (CRs). Both CRs and extracellular levels of these three amino acids returned to control values during extinction. Other amino acids (alanine, GABA, glutamine, serine, and threonine) did not undergo modifications in their extracellular concentrations across the training. Results are discussed with regard to the role of PIN in this type of associative learning. PMID- 17907843 TI - Previous experience affects subsequent anxiety-like responses in rats bred for novelty seeking. AB - Novelty-seeking behavior in rats is deemed to model sensation seeking in humans, a personality trait related to some psychiatric conditions, including substance abuse. Animals characterized based on their locomotor response to novelty, namely high and low responders (HRs and LRs, respectively), show differences in anxiety and drug-taking behaviors. This study evaluates the effect of anxiety-provoking situations on subsequent behaviors in these endophenotypes. Selectively bred HR and LR rats were submitted to blocks of tests consisting of two-trial light- dark (LD) and two-trial elevated plus maze (EPM) tests arranged in counterbalanced, alternating order. No differences in anxiety-like behaviors were found in HR-bred and LR-bred rats in either LD trial, regardless of the test order. Repeated exposure to the EPM test, however, resulted in enhanced behavioral response under different test orders as a function of endophenotype. Compared with HR-bred animals, LR-bred animals exhibited increased anxiety on reexposure to EPM but only if both trials were preceded by an LD test session. The emotional responses in HR-bred and LR-bred rats reported here may reflect different degrees of adaptive processing regulated by both genetic and environmental factors. PMID- 17907844 TI - Extracellular signal-regulated kinase 1/2 involvement in the enhancement of contextual fear conditioning by nicotine. AB - Contextual fear conditioning is enhanced by nicotine, but the cellular mechanisms underlying this effect are unknown. Extracellular signal regulated kinase 1/2 (ERK 1/2) has been shown to play an integral role in the formation of contextual fear memories. As such, it is possible that ERK 1/2 is involved in the enhancement of contextual fear conditioning by nicotine. To determine whether ERK 1/2 plays a role in this enhancement, a dose of SL327 (a selective, systemic ERK 1/2 inhibitor) that is subthreshold for inhibiting contextual fear conditioning was coadministered with nicotine prior to training, testing, or both training and testing of contextual fear conditioning in C57BL/6 mice. When administered prior to training, this subthreshold dose of SL327 attenuated the enhancement of contextual fear conditioning by nicotine to levels similar to those of vehicle treated animals. When administered prior to testing, the subthreshold dose of SL327 did not significantly alter conditioning. These results suggest that activation of ERK 1/2 by nicotine during acquisition leads to an enhancement of contextual fear conditioning. PMID- 17907845 TI - Systemic or intrahippocampal delivery of histone deacetylase inhibitors facilitates fear extinction. AB - Several recent studies have shown that chromatin, the DNA-protein complex that packages genomic DNA, has an important function in learning and memory. Dynamic chromatin modification via histone deacetylase (HDAC) inhibitors and histone acetyltransferases may enhance hippocampal synaptic plasticity and hippocampus dependent memory. Little is known about the effects of HDAC inhibitors on extinction, a learning process through which the ability of a previously conditioned stimulus, such as a conditioning context, to evoke a conditioned response is diminished. The authors demonstrate that administration of the HDAC inhibitors sodium butyrate (NaB) systemically or trichostatin A (TSA) intrahippocampally prior to a brief (3-min) contextual extinction session causes context-evoked fear to decrease to levels observed with a long (24-min) extinction session. These results suggest that HDAC inhibitors may enhance learning during extinction and are consistent with other studies demonstrating a role for the hippocampus in contextual extinction. Molecular and behavioral mechanisms through which this enhanced extinction effect may occur are discussed. PMID- 17907846 TI - Binary mixture perception is affected by concentration of odor components. AB - Some controversy still exists as to how binary odorant mixtures are behaviorally perceived, despite many studies aimed at understanding this phenomenon. Binary mixture perception by rodents is a first step in elucidating how more complex odor blends may be perceived. Research thus far has examined how the degree of component similarity, olfactory receptor overlap, relative concentration of components, and even olfactory enrichment affect the behavioral perception of binary mixtures. These studies have aimed to categorize binary mixtures into 1 of 3 rigid categories, but often the results conflict as to which category a particular mixture belongs. In the present article, the authors used a habituation/discrimination paradigm to determine whether rats' perception of one component of a binary mixture of either perceptually similar or dissimilar components changed when the concentration of both components was varied together. The authors found that perception of a binary mixture changed with changing component concentration, such that one binary mixture could be categorized differently depending on component intensity. PMID- 17907847 TI - Supersmart mice: surprising or surprised? Theoretical comment on Singer, Boison, Mohler, Feldon, and Yee (2007). AB - The glycine transporter (GlyT1) regulates levels of the neurotransmitter glycine, a coagonist of the N-methyl-D-aspartate receptor (NMDAR), and as such may represent a novel site for developing cognition-enhancing drugs. Genetically modified mice with reduced levels of GlyT1 have been generated to test this hypothesis. P. Singer, D. Boison, H. Mohler, J. Feldon, and B. K. Yee now show, through a spontaneous exploration task, that mice in which GlyT1 has been deleted, specifically in neurons in the forebrain, demonstrate enhanced object recognition memory. Whereas both control and mutant mice show a preference for a novel object over a familiar object 2 min after the initial presentation of 1 of the objects, only the mutant mice show a preference for the novel object when tested after a 2-hr delay. The longer-lasting habituation displayed by the GlyT1 mice is consistent with a role for glycine/NMDAR-dependent synaptic plasticity in supporting a nonassociative, short-term memory trace of a recently experienced stimulus. This short-term habituation process may be independent of associative learning mechanisms and may be best described by A. R. Wagner's (1981) sometimes opponent process model. PMID- 17907848 TI - Cellular learning theory: theoretical comment on Cole and McNally (2007). AB - The idea that learning proceeds as a function of the discrepancy (or error) between expected and obtained outcomes is central to many theories of associative learning. However, remarkably little is known about the neurobiological mechanisms that underlie this learning of predictive errors in fear conditioning, a widely used preparation in studies of cellular and molecular mechanisms of memory. In this issue of Behavioral Neuroscience, S. Cole and G. P. McNally demonstrate an important dissociation between the establishment and regulation of predictive error at the cellular level. Their findings have added a level of complexity to currently established views of the function of NMDA and opioid receptors in learning and memory. This commentary discusses some of the implications of these findings for theoretical and neurobiological approaches to memory, as well as current thinking about the cellular circuitry involved in reward learning and drug abuse. PMID- 17907849 TI - What predicts psychological resilience after disaster? The role of demographics, resources, and life stress. AB - A growing body of evidence suggests that most adults exposed to potentially traumatic events are resilient. However, research on the factors that may promote or deter adult resilience has been limited. This study examined patterns of association between resilience and various sociocontextual factors. The authors used data from a random-digit-dial phone survey (N = 2,752) conducted in the New York City area after the September 11, 2001, terrorist attack. Resilience was defined as having 1 or 0 posttraumatic stress disorder symptoms and as being associated with low levels of depression and substance use. Multivariate analyses indicated that the prevalence of resilience was uniquely predicted by participant gender, age, race/ethnicity, education, level of trauma exposure, income change, social support, frequency of chronic disease, and recent and past life stressors. Implications for future research and intervention are discussed. PMID- 17907850 TI - Predictors of posttraumatic distress 1 year after exposure to community violence: the importance of acute symptom severity. AB - In this longitudinal study of 333 primarily male, Hispanic survivors of community violence, the authors investigated the effects of 4 categories of risk factors on posttraumatic stress disorder (PTSD) symptom severity: demographic characteristics, pretraumatic psychological factors, characteristics of the trauma, and reactions to the trauma. Replicating past research, exemplars from all 4 categories predicted PTSD symptom severity at 12-month follow-up. Acute symptom severity, measured approximately 5 days posttrauma, accounted for the largest proportion of variance among all the predictors included. No other predictors remained significant after 5-day distress was included in the model. These findings suggest that the effects of several purported risk factors for chronic posttraumatic distress may already be reflected in acute distress following trauma exposure. These results bear on current conceptions of the fundamental nature of PTSD and suggest that initial distress during the immediate aftermath of the trauma may be an important target for intervention. PMID- 17907851 TI - Prevention of depressive symptoms in adolescents: a randomized trial of cognitive behavioral and interpersonal prevention programs. AB - This study evaluated the efficacy of 2 programs for preventing depressive symptoms in adolescents. Participants were 380 high school students randomly assigned to a cognitive-behavioral program (CB), an interpersonal psychotherapy adolescent skills training program (IPT-AST), or a no-intervention control. The interventions involved eight 90-min weekly sessions run in small groups during wellness classes. At postintervention, students in both the CB and IPT-AST groups reported significantly lower levels of depressive symptoms than did those in the no-intervention group, controlling for baseline depression scores; the 2 intervention groups did not differ significantly from each other. The effect sizes, using Cohen's d, for the CB intervention and the IPT-AST intervention were 0.37 and 0.26, respectively. Differences between control and intervention groups were largest for adolescents with high levels of depressive symptoms at baseline. For a high-risk subgroup, defined as having scored in the top 25th percentile on the baseline depression measure, the effect sizes for the CB and the IPT-AST interventions were 0.89 and 0.84, respectively. For the whole sample, sociotropy and achievement orientation moderated the effect of the interventions. Intervention effects were short term and were not maintained at 6-month follow up. PMID- 17907854 TI - Empirically derived subtypes of adolescent depression: latent profile analysis of co-occurring symptoms in the Treatment for Adolescents with Depression Study (TADS). AB - A latent profile analysis was conducted on the co-occurring symptoms of 423 adolescents diagnosed with major depressive disorder as part of the Treatment for Adolescents with Depression Study (TADS), a multisite, randomized treatment trial. The participants had a mean (SD) age of 14.6 (1.5) years; of the sample, 45.6% was male and 73.8% was white. Scores on the primary subscales of Conners' Parent Rating Scale--Revised were used as indicators in the analysis. Five classes of symptoms best described the clinical presentation of adolescents enrolled in the TADS. Of the adolescents in the sample, 80% were assigned to classes with clinically significant elevations on 1 or more subscales of the Children's Depression Rating Scale--Revised. The 5 classes met empirical criteria for distinctiveness and were validated against clinical diagnoses, child-rated symptoms, and clinician-rated functional impairment. The findings are consistent with prior studies that showed a high rate of co-occurring symptoms among depressed adolescents. The discussion focuses on understanding subtypes and comorbidity in adolescent depression as well as the implications for treatment and for prevention. PMID- 17907852 TI - Prediction of suicide ideation and attempts among adolescents using a brief performance-based test. AB - Suicide is a leading cause of death that is difficult to predict because clinical assessment has relied almost exclusively on individuals' self-report of suicidal thoughts. This is problematic because there often is motivation to conceal such thoughts. The authors tested the ability of the Self-Injury Implicit Association Test (SI-IAT), a reaction-time measure of implicit associations between self injury and oneself, to detect and predict suicide ideation and attempts. Participants were adolescents who were nonsuicidal (n = 38), suicide ideators (n = 37), or recent suicide attempters (n = 14). Analyses revealed large between group differences on the SI-IAT, with nonsuicidal adolescents showing large negative associations between self-injury and themselves, suicide ideators showing small positive associations, and suicide attempters showing large positive associations on this performance-based test. The SI-IAT accurately predicted current suicide ideation and attempt status as well as future suicide ideation, and it incrementally improved prediction of these outcomes above and beyond the use of known risk factors. Future research is needed to refine this assessment method and to further develop and examine performance-based assessment of suicide risk in clinical settings. PMID- 17907855 TI - The generalizability of the Youth Self-Report syndrome structure in 23 societies. AB - As a basis for theories of psychopathology, clinical psychology and related disciplines need sound taxonomies that are generalizable across diverse populations. To test the generalizability of a statistically derived 8-syndrome taxonomic model for youth psychopathology, confirmatory factor analyses (CFAs) were performed on the Youth Self-Report (T. M. Achenbach & L. A. Rescorla, 2001) completed by 30,243 youths 11-18 years old from 23 societies. The 8-syndrome taxonomic model met criteria for good fit to the data from each society. This was consistent with findings for the parent-completed Child Behavior Checklist (Achenbach & Rescorla, 2001) and the teacher-completed Teacher's Report Form (Achenbach & Rescorla, 2001) from many societies. Separate CFAs by gender and age group supported the 8-syndrome model for boys and girls and for younger and older youths within individual societies. The findings provide initial support for the taxonomic generalizability of the 8-syndrome model across very diverse societies, both genders, and 2 age groups. PMID- 17907856 TI - Multivariate models of mothers' and fathers' aggression toward their children. AB - Multivariate, biopsychosocial, explanatory models of mothers' and fathers' psychological and physical aggression toward their 3- to 7-year-old children were fitted and cross-validated in 453 representatively sampled families. Models explaining mothers' and fathers' aggression were substantially similar. Surprisingly, many variables identified as risk factors in the parental aggression and physical child abuse literatures, such as income, unrealistic expectations, and alcohol problems, although correlated with aggression bivariately, did not contribute uniquely to the models. In contrast, a small number of variables (i.e., child responsible attributions, overreactive discipline style, anger expression, and attitudes approving of aggression) appeared to be important pathways to parent aggression, mediating the effects of more distal risk factors. Models accounted for a moderate proportion of the variance in aggression. PMID- 17907857 TI - Multivariate models of men's and women's partner aggression. AB - This exploratory study was designed to address how multiple factors drawn from varying focal models and ecological levels of influence might operate relative to each other to predict partner aggression, using data from 453 representatively sampled couples. The resulting cross-validated models predicted approximately 50% of the variance in men's and women's partner aggression. The 3 strongest direct predictors of partner aggression for men and women were dominance/jealousy, marital adjustment, and partner responsibility attributions. Three additional direct paths to aggression for men were exposure to family-of-origin aggression, anger expression, and perceived social support. The 1 additional direct path for women was a history of their own aggression as a child or teenager. Implications for more integrative theories and intervention are discussed. PMID- 17907858 TI - Randomized trial of prize-based reinforcement density for simultaneous abstinence from cocaine and heroin. AB - To examine the effect of reinforcer density in prize-based abstinence reinforcement, heroin/cocaine users (N = 116) in methadone maintenance (100 mg/day) were randomly assigned to a noncontingent control group (NonC) or to 1 of 3 groups that earned prize draws for abstinence: manual drawing with standard prize density (MS) or computerized drawing with standard (CS) or high (CH) density. Probabilities (prizes/draw) were standard (50%) and high (78%); prize density was double blind. Mean prize values were CH, $286; CS, $167; MS, $139; and NonC, $171. Outcomes were % opioid/cocaine-negative urines during the 12-week intervention and then 8 weeks postintervention as well as diagnosis of dependence up to 6 months poststudy. CH had significantly more negative specimens than did NonC during intervention and had more than all groups during postintervention treatment: Mean % negative (95% confidence interval) during postintervention treatment adjusted for baseline drug use and dropout were CH, 55% (14%-90%); CS, 7% (1%-27%); MS, 4% (1%-12%); and NonC, 3% (1%-10%). Current cocaine dependence diagnoses after treatment were significantly lower in contingent compared with noncontingent groups. Computerized drawing with higher-density prizes enhanced reduction of cocaine use; abstinence reinforcement had long-term therapeutic benefits. PMID- 17907859 TI - Mediators of telephone-based continuing care for alcohol and cocaine dependence. AB - A previous randomized trial with 224 alcohol and/or cocaine addicts who had completed an initial phase of treatment indicated that 12 weeks of telephone based continuing care yielded higher abstinence rates over 24 months than did group counseling continuing care. The current study examined mediators of this treatment effect. Results suggested that self-help involvement during treatment and self-efficacy and commitment to abstinence 3 months after treatment mediated subsequent abstinence outcomes. These analyses controlled for substance use prior to the assessment of mediators. Conversely, there was no evidence that self-help beliefs or social support mediated the treatment effect. These results are consistent with a model in which treatment effects are first accounted for by changes in behavior, followed by changes in self-efficacy and in commitment to abstinence. PMID- 17907861 TI - The epidemiology of psychiatric disorders among repeat DUI offenders accepting a treatment-sentencing option. AB - Psychiatric comorbidity likely contributes to driving under the influence (DUI) of alcohol among repeat offenders. This study presents one of the first descriptions of the prevalence and comorbidity of psychiatric disorders among repeat DUI offenders in treatment. Participants included all consenting eligible admissions (N = 729) to a 2-week inpatient treatment facility for court-sentenced repeat DUI offenders (i.e., offenders electing treatment in place of prison time) from April 17, 2005, to April 23, 2006. Participants completed the Composite International Diagnostic Interview, which assessed the following disorders using criteria from the Diagnostic and Statistical Manual of Mental Disorders (4th ed.; American Psychiatric Association, 1994): alcohol use and drug use, bipolar, generalized anxiety, posttraumatic stress, intermittent explosive, conduct, attention deficit, nicotine dependence, pathological gambling, and major depressive. Repeat DUI offenders evidenced higher lifetime and 12-month prevalence of alcohol use and drug use disorders, conduct disorder, posttraumatic stress disorder, generalized anxiety disorder, and bipolar disorder compared with the general population. Almost half qualified for lifetime diagnoses of both addiction (i.e., alcohol, drug, nicotine, and/or gambling) and a psychiatric disorder. Lifetime and past-year comorbidity rates were higher among participants than in the general population. These results suggest that clinicians should consider multimorbidity within DUI treatment protocols. PMID- 17907860 TI - Mental health and substance use disorders among Latino and Asian American lesbian, gay, and bisexual adults. AB - Growing evidence suggests that lesbian, gay, and bisexual adults may be at elevated risk for mental health and substance use disorders, possibly due to anti gay stigma. Little of this work has examined putative excess morbidity among ethnic/racial minorities resulting from the experience of multiple sources of discrimination. The authors report findings from the National Latino and Asian American Survey (NLAAS), a national household probability psychiatric survey of 4,488 Latino and Asian American adults. Approximately 4.8% of persons interviewed identified as lesbian, gay, bisexual, and/or reported recent same-gender sexual experiences. Although few sexual orientation-related differences were observed, among men, gay/bisexual men were more likely than heterosexual men to report a recent suicide attempt. Among women, lesbian/bisexual women were more likely than heterosexual women to evidence positive 1-year and lifetime histories of depressive disorders. These findings suggest a small elevation in psychiatric morbidity risk among Latino and Asian American individuals with a minority sexual orientation. However, the level of morbidity among sexual orientation minorities in the NLAAS appears similar to or lower than that observed in population-based studies of lesbian, gay, and bisexual adults. PMID- 17907862 TI - Effectiveness of abstinence-based incentives: interaction with intake stimulant test results. AB - Intake urinalysis test result (drug positive vs. negative) has been previously identified as a strong predictor of drug abuse treatment outcome, but there is little information about how this prognostic factor may interact with the type of treatment delivered. The authors used data from a multisite study of abstinence incentives for stimulant abusers enrolled in outpatient counseling treatment (N. M. Petry, J. M. Peirce, et al., 2005) to examine this question. The first study urine was used to stratify participants into stimulant negative (n = 306) versus positive (n = 108) subgroups. Abstinence incentives significantly improved retention in those testing negative but not in those testing positive. Findings suggest that stimulant abusers presenting to treatment with a stimulant-negative urine benefit from abstinence incentives, but alternative treatment approaches are needed for those who test stimulant positive at intake. PMID- 17907863 TI - Maladaptive self-appraisals before trauma exposure predict posttraumatic stress disorder. AB - This study tested the proposal that negative appraisals represent a risk factor for developing posttraumatic stress disorder (PTSD) after trauma. Trainee firefighters (N = 68) were assessed during training (before trauma exposure) for PTSD, history of traumatic events, and tendency to engage in negative appraisals. Firefighters were reassessed 4 years later (N = 52), after commencing firefighter duty (after trauma exposure), for PTSD and depression using the Posttraumatic Stress Disorder Scale (E. B. Foa, L. Cashman, L. Jaycox, & K. Perry, 1997) and the Beck Depression Inventory (Version 2; A. T. Beck, R. A. Steer, & G. K. Brown, 1996). At follow-up, 12% met criteria for PTSD. Pretrauma negative appraisals about oneself accounted for 20% of variance in PTSD severity at follow-up. These data provide the first evidence that preexisting negative appraisals are a risk factor for PTSD. PMID- 17907864 TI - Efficacy of group motivational interviewing (GMI) for psychiatric inpatients with chemical dependence. AB - Dually diagnosed patients with chemical dependency and a comorbid psychiatric disorder typically show poor compliance with aftercare treatment, which may result in costly and pervasive individual and societal problems. In this study, the authors investigated the effect of adding motivational interviewing in a group format to standard treatment for dually diagnosed psychiatric inpatients. The patients (n = 101) all received standard care and in addition were assigned to either group motivational interviewing (GMI) or a therapist attention activity control group (TAAC). Of patients who attended aftercare and who used alcohol or drugs, those who participated in GMI attended significantly more aftercare treatment sessions, consumed less alcohol, and engaged in less binge drinking at follow-up compared with those in TAAC. Differences between conditions in the overall percentage of participants who achieved complete abstinence or who attended aftercare treatment were not significant, possibly because of a lack of power. These results provide preliminary evidence for the efficacy of GMI when added at the outset to an inpatient program. PMID- 17907865 TI - Meta-analysis of day treatment and contingency-management dismantling research: Birmingham Homeless Cocaine Studies (1990-2006). AB - Four successive randomized clinical trials studying contingency management (CM), involving various treatment arms of drug-abstinent housing and work therapy and day treatment (DT) with a behavioral component, were compared on common drug abstinence outcomes at 2 treatment completion points (2 and 6 months). The clinical trials were conducted from 1990 to 2006 in Birmingham, Alabama, with a total of 644 homeless persons with primary crack cocaine addiction. The meta analysis utilized the weighted least squares approach to integrate data encompassing 9 different treatment arms to assess the effects of CM and DT (neither, DT only, CM only, and CM = DT) on a common estimate of prevalence of drug abstinence. Taken together, the results show much stronger benefits from CM = DT and from CM only than for DT alone. Throughout all of the Birmingham Homeless Cocaine Studies, the CM = DT consistently produced higher abstinence prevalence than did no CM. PMID- 17907866 TI - A new look at habits and the habit-goal interface. AB - The present model outlines the mechanisms underlying habitual control of responding and the ways in which habits interface with goals. Habits emerge from the gradual learning of associations between responses and the features of performance contexts that have historically covaried with them (e.g., physical settings, preceding actions). Once a habit is formed, perception of contexts triggers the associated response without a mediating goal. Nonetheless, habits interface with goals. Constraining this interface, habit associations accrue slowly and do not shift appreciably with current goal states or infrequent counterhabitual responses. Given these constraints, goals can (a) direct habits by motivating repetition that leads to habit formation and by promoting exposure to cues that trigger habits, (b) be inferred from habits, and (c) interact with habits in ways that preserve the learned habit associations. Finally, the authors outline the implications of the model for habit change, especially for the self regulation of habit cuing. PMID- 17907867 TI - On seeing human: a three-factor theory of anthropomorphism. AB - Anthropomorphism describes the tendency to imbue the real or imagined behavior of nonhuman agents with humanlike characteristics, motivations, intentions, or emotions. Although surprisingly common, anthropomorphism is not invariant. This article describes a theory to explain when people are likely to anthropomorphize and when they are not, focused on three psychological determinants--the accessibility and applicability of anthropocentric knowledge (elicited agent knowledge), the motivation to explain and understand the behavior of other agents (effectance motivation), and the desire for social contact and affiliation (sociality motivation). This theory predicts that people are more likely to anthropomorphize when anthropocentric knowledge is accessible and applicable, when motivated to be effective social agents, and when lacking a sense of social connection to other humans. These factors help to explain why anthropomorphism is so variable; organize diverse research; and offer testable predictions about dispositional, situational, developmental, and cultural influences on anthropomorphism. Discussion addresses extensions of this theory into the specific psychological processes underlying anthropomorphism, applications of this theory into robotics and human-computer interaction, and the insights offered by this theory into the inverse process of dehumanization. PMID- 17907868 TI - A neural network model of retrieval-induced forgetting. AB - Retrieval-induced forgetting (RIF) refers to the finding that retrieving a memory can impair subsequent recall of related memories. Here, the authors present a new model of how the brain gives rise to RIF in both semantic and episodic memory. The core of the model is a recently developed neural network learning algorithm that leverages regular oscillations in feedback inhibition to strengthen weak parts of target memories and to weaken competing memories. The authors use the model to address several puzzling findings relating to RIF, including why retrieval practice leads to more forgetting than simply presenting the target item, how RIF is affected by the strength of competing memories and the strength of the target (to-be-retrieved) memory, and why RIF sometimes generalizes to independent cues and sometimes does not. For all of these questions, the authors show that the model can account for existing results, and they generate novel predictions regarding boundary conditions on these results. PMID- 17907870 TI - Sophisticated approval voting, ignorance priors, and plurality heuristics: a behavioral social choice analysis in a Thurstonian framework. AB - This project reconciles historically distinct paradigms at the interface between individual and social choice theory, as well as between rational and behavioral decision theory. The authors combine a utility-maximizing prescriptive rule for sophisticated approval voting with the ignorance prior heuristic from behavioral decision research and two types of plurality heuristics to model approval voting behavior. When using a sincere plurality heuristic, voters simplify their decision process by voting for their single favorite candidate. When using a strategic plurality heuristic, voters strategically focus their attention on the 2 front-runners and vote for their preferred candidate among these 2. Using a hierarchy of Thurstonian random utility models, the authors implemented these different decision rules and tested them statistically on 7 real world approval voting elections. They cross-validated their key findings via a psychological Internet experiment. Although a substantial number of voters used the plurality heuristic in the real elections, they did so sincerely, not strategically. Moreover, even though Thurstonian models do not force such agreement, the results show, in contrast to common wisdom about social choice rules, that the sincere social orders by Condorcet, Borda, plurality, and approval voting are identical in all 7 elections and in the Internet experiment. PMID- 17907869 TI - The fSAM model of false recall. AB - The authors report a new theory of false memory building upon existing associative memory models and implemented in fSAM, the first fully specified quantitative model of false recall. Participants frequently intrude unstudied critical words while recalling lists comprising their strongest semantic associates but infrequently produce other extralist and prior-list intrusions. The authors developed the theory by simulating recall of such lists, using factorial combinations of semantic mechanisms operating at encoding, retrieval, or both stages. During encoding, unstudied words' associations to list context were strengthened in proportion to their strength of semantic association either to each studied word or to all co-rehearsed words. During retrieval, words received preference in proportion to their strength of semantic association to the most recently recalled single word or multiple words. The authors simulated all intrusion types and veridical recall for lists varying in semantic association strength among studied and critical words from the same and different lists. Multiplicative semantic encoding and retrieval mechanisms performed well in combination. Using such combined mechanisms, the authors also simulated several core findings from the Deese-Roediger-McDermott paradigm literature, including developmental patterns, specific list effects, association strength effects, and true-false correlations. These results challenge existing false memory theories. PMID- 17907871 TI - Exploiting redundancy for flexible behavior: unsupervised learning in a modular sensorimotor control architecture. AB - Autonomously developing organisms face several challenges when learning reaching movements. First, motor control is learned unsupervised or self-supervised. Second, knowledge of sensorimotor contingencies is acquired in contexts in which action consequences unfold in time. Third, motor redundancies must be resolved. To solve all 3 of these problems, the authors propose a sensorimotor, unsupervised, redundancy-resolving control architecture (SURE_REACH), based on the ideomotor principle. Given a 3-degrees-of-freedom arm in a 2-dimensional environment, SURE_REACH encodes 2 spatial arm representations with neural population codes: a hand end-point coordinate space and an angular arm posture space. A posture memory solves the inverse kinematics problem by associating hand end-point neurons with neurons in posture space. An inverse sensorimotor model associates posture neurons with each other action-dependently. Together, population encoding, redundant posture memory, and the inverse sensorimotor model enable SURE_REACH to learn and represent sensorimotor grounded distance measures and to use dynamic programming to reach goals efficiently. The architecture not only solves the redundancy problem but also increases goal reaching flexibility, accounting for additional task constraints or realizing obstacle avoidance. While the spatial population codes resemble neurophysiological structures, the simulations confirm the flexibility and plausibility of the model by mimicking previously published data in arm-reaching tasks. PMID- 17907872 TI - Stimulus-dependent dopamine release in attention-deficit/hyperactivity disorder. AB - Attention-deficit/hyperactivity disorder (ADHD) is related to an attenuated and dysfunctional dopamine system. Normally, a high extracellular dopamine level yields a tonic dopaminergic input that down-regulates stimuli-evoked phasic dopamine responses through autoreceptors. Abnormally low tonic extracellular dopamine in ADHD up-regulates the autoreceptors so that stimuli-evoked phasic dopamine is boosted. The authors propose that these boosted phasic responses yield hypersensitivity to environmental stimuli in ADHD. Stimuli evoking moderate brain arousal lead to well-functioning performance, whereas either too little or too much stimuli attenuate cognitive performance. Strong, salient stimuli may easily disrupt attention, whereas an environment with impoverished stimuli causes low arousal, which is typically compensated for by hyperactivity. Stochastic resonance is the phenomenon that makes a moderate noise facilitate stimulus discrimination and cognitive performance. Computational modeling shows that more noise is required for stochastic resonance to occur in dopamine-deprived neural systems in ADHD. This prediction is supported by empirical data. PMID- 17907873 TI - Item-specific adaptation and the conflict-monitoring hypothesis: a computational model. AB - M. M. Botvinick, T. S. Braver, D. M. Barch, C. S. Carter, and J. D. Cohen (2001) implemented their conflict-monitoring hypothesis of cognitive control in a series of computational models. The authors of the current article first demonstrate that M. M. Botvinick et al.'s (2001) conflict-monitoring Stroop model fails to simulate L. L. Jacoby, D. S. Lindsay, and S. Hessels's (2003) report of an item specific proportion-congruent (ISPC) effect in the Stroop task. The authors then implement a variant of M. M. Botvinick et al.'s model based on the assumption that control must be able to operate at the item level. This model successfully simulates the ISPC effect. In addition, the model provides an alternative to M. M. Botvinick et al.'s explanation of the list-level proportion-congruent effect in terms of an ISPC effect. Implications of the present modeling effort are discussed. PMID- 17907874 TI - Sortal concepts and causal continuity: comment of Rips, Blok, and Newman (2006). AB - L. J. Rips, S. Blok, and G. Newman (2006) proposed that singular concepts, which support the tracing of individual objects across their existence, are governed by a principle of causal continuity. They purported to show that causal continuity is better than existing theories at explaining judgments of the persistence of individual objects. This article makes 3 points. First, the construct of causal connectedness entails a wide variety of different explanatory factors; calling them all causal has questionable explanatory value. There is little evidence that the ultimate basis for identity judgments in many cases is causal. Second, the authors suggest that causal knowledge is indeed important to identity, but that it is important in the context of sortal concepts; different causal information matters for different kinds of things. Finally, the authors consider whether causal knowledge or sortal concepts are more fundamental to tracing individual identity, that is, whether causal knowledge is necessary for identity judgments. The authors appeal to research in developmental psychology that has begun to address this debate, supporting the primacy of sortal concepts. Although there continues to be shortcomings of all theories of object persistence, it is not clear that the causal continuer theory brings new clarity to the puzzle. PMID- 17907875 TI - Postscript: sortal concepts are fundamental for tracing identity. AB - Our discussion with Rips, Blok, and Newman (2006; Blok, Newman, & Rips, in press; Rhemtulla & Xu, in press) has brought together theory and evidence from researchers in adult cognition and those in infant development. By this point, researchers all recognize that any theory of concepts of individuals must consider the evidence from infants' first concepts as well as explicit adult reasoning tasks. Despite this agreement, we continue to disagree on several fundamental issues. PMID- 17907879 TI - The double-anchoring theory of lightness perception: a comment on Bressan (2006). AB - Recently, a double-anchoring theory (DAT) of lightness perception was proposed (P. Bressan, 2006), which offers explanations for all the data explained by the original anchoring theory (A. Gilchrist et al., 1999), as well as a number of additional lightness phenomena. Consequently, DAT can account for an unprecedented range of empirical results, potentially explaining everything from the basic simultaneous contrast display to subtle variations of the Gelb effect. In this comment, the authors raised 4 concerns that demonstrate serious theoretical and empirical difficulties for DAT. PMID- 17907883 TI - [Evolution of anxiety and depression detected during hospitalization in an Internal Medicine service]. PMID- 17907884 TI - [Evolution of anxiety and depression detected during hospitalization in an Internal Medicine service]. AB - OBJECTIVE: To study the evolution of the symptoms of anxiety (Ax) and depression (Dp) two months after hospital release and factors associated with their persistence. METHOD: An observational longitudinal study. Patients with Ax or Dp, evaluated by the Hospital Anxiety and Depression (HAD) scale on the fourth day of hospitalisation, were followed over two months without the use of psychopharmacological drugs. Sociodemographic factors, psychiatric history, functional state, comorbidity and the HAD stage three weeks before admission were evaluated. RESULTS: Thirty-eight patients with Ax (22 males) with a mean age of 62.2 years and 35 patients with Dp (22 males) with a mean age of 68.1 years were studied. Symptoms of Ax persisted in 23 patients (60.5%; CI95%: 43.4-76) and symptoms of Dp persisted in 18 (51.4%; CI95%: 34-68.6). Ax before admission and a lower educational level were associated with the persistence of Ax. An age of 70 or more, female gender and primary studies, Barthel Index on admission of < 100 and depression in the three weeks before admission were associated with persistence of Dp. CONCLUSIONS: Symptoms persisted in over half of the patients. The evaluation of HAD before admission may help in determining the treatment course to be followed. PMID- 17907885 TI - [Prognosis of patients who recovered after an episode of sudden death]. AB - OBJECTIVES: Sudden death constitutes a major sanitary problem with high mortality and serious neurological complications. The objective of this study was to analyze the prognosis and the characteristics of patients who initially recovered after an episode of cardiac arrest and who were admitted to the intensive care unit (ICU). METHOD: We retrospectively studied the clinical characteristics and outcome of 65 patients admitted to the Intensive Care Unit during a 3 years period with aborted sudden death. RESULTS: 65 patients, 44 (67.7%) men and 21 (32.3%) women. Middle ages 69.1 +/- 13.9. 29 (44.6%) out of hospital and 36 (55.4%) into hospital sudden death. Cardiopulmonary resuscitation was <10 minutes in 28 cases (43.1%), and > 10 minutes in 37 (56.9%). 36 (55.4%) of all sudden deaths were of cardiac origin. 37 patients (56.9%) died and 28 (43.1%) survived the episode. It was LET in 11 cases (16.9%). 29 (44.6%) of all had post- anoxic encephalopathy and most died before discharge from ICU. Of 28 survivors, 5 patients were discharged alive with post-anoxic encephalopathy (17.8%) and 23 were discharged without neurological disturbances (82.2%). This was more frequent when sudden death was into hospital (p 0.009) and cardiopulmonary resuscitation was < 10 minutes (p 0.045). CONCLUSIONS: High number of the patients admitted to a Intensive Care unit with aborted sudden death died during ICU stay. Many patients had post-anoxic encephalopathy and most of these died. So, up to 35% of the patients admitted after an episode of cardiac arrest were discharged alive and without severe neurological damage. PMID- 17907886 TI - [Hidden renal failure and drug prescription in hospitalized patients]. AB - OBJECTIVE: Equations used to estimate glomerular filtarion rate (GFR) are useful to detect "hidden" renal failure (RF) ( coexistence of normal serum creatinine together with GFR < 60 ml/mto/1.73 m2 ). The aim of this study was to analyze the prevalence of this phenomenon in hospitalized patients and how this affects drugs prescription. PATIENTS AND METHOD: 567 patients admitted to the internal medicine section of Barbastro Hospital (Huesca, Spain) during a three months period (april june 2006) were included. GFR was estimated applying abbreviated MDRD equation. By means of logistic regression analysis we analyzed the factors associated with the presence of "hidden" RF. We also recorded the potentially dangerous drugs prescribed at discharge from hospital. RESULTS: Prevalence of "Hidden" RF was 10.8 %. NKF chronic kidney disease stages 3.4 or 5 were present in 35.2 % of patients. Variables associated with presence of "hidden" RF were age (expB = 1.035; IC95% 1.012 - 1.060; p = 0.004) and female sex (expB = 4.669; IC95% 2.461 8.856; p < 0.0001). Only in 1.8 % of cases GFR was calculated during hospitalisation period. A considerable number of patients with "hidden" RF received treatment with potentially dangerous drugs taking into account his degree of renal failure. CONCLUSIONS: Our results support the introduction of GFR estimation by means of MDRD equation in laboratory reports. PMID- 17907887 TI - [Fluoroquinolone-induced tendon diseases]. AB - Tendinitis and tendon ruptures induced by fluoroquinolones, while uncommon, have been documented in the literature since 1983. We report five cases of tendinitis induced by fluoroquinolones, three caused by levofloxacin and two by ciprofloxacin. We revise actual knowledge of this association and we insist on how important is an early detection to prevent tendon rupture. PMID- 17907888 TI - [Splenic irradiation: apropos of 8 cases. Clinical indications and literature review]. AB - Clinical indications of splenic irradiation in haematological disorders include the irradiation in lymphoproliferative disorders with spleen infiltration, palliative treatment of splenomegaly in malignant diseases like chronic lymphocytic leukaemia or myeloproliferative disorders, with the purpose of relief from abdominal pain associated with capsular enlargement size and decrease cytopenias secundaries to hypersplenism.This paper reports our experience with spleen irradiation in the Hospital General Universitario Gregorio Maranon in the last five years, and analyzes indications, results and toxicity, and an actual review of the literature. PMID- 17907890 TI - [Lhermitte-Duclos's disease associated to Cowden s disease: a case report]. AB - Cowden's disease is a rare genodermatosis that is characterized for multiple cutaneous and visceral hamartoma . Lhermitte-Duclos's disease is a cerebelous lesion that consists in the displasic enlargement of the cerebelous circumvolution. It's incluyed in phacomatosis and usually presents associated to Cowden's disease, tuberous sclerosis and overlap syndromes.A 56 years old man, diagnosed in Dermatology with Cowden's disease ten years ago. In the extension study, he had hamartoma intestinal polip, esophagic glucogenic acanthosis and two solid thyroid nodules. The craneal TC didn't show significant alteration. Ten years after diagnosis a cerebral magnetis resonance was performed for intense cephalea, and it was found a bad-defined mass in right cerebelous hemisphere without contrast captation, compatible with cerebelous glangliocytoma. PMID- 17907889 TI - [Acute renal failure due to sulfadiazine crystalluria]. AB - Focal necrotizing encephalitis due to Toxoplasma gondii infection represents one of the most common opportunistic infection in patients with the acquired inmunodeficiency syndrome (AIDS), and the treatment is commonly with a combination sulphadiazine, and pyrimethamine. A major side effect of sulfadiazine therapy is the occurrence of crystallization in the urinary collecting system. We report a patient with AIDS and Toxoplasmic encephalitis treated with sulfadiazine who developed acute renal failure. Renal ultrasound demonstrated echogenic areas within the renal parenchyma, presumed to be sulfa crystals. Renal failure and ultrasound findings resolved rapidly with hydratation and administration of alkali. Patients infected with AIDS frequently have characteristic that increase intratubular crystal precipitation and they require treatment with one or more of the drugs that are associated with crystal-induced renal failure. Controlled alkalinization of the urine and high fluid intake are recommended for prophylaxis of crystalluria. The literature concerning crystalluria and renal failure due to sulfadiazine is reviewed. PMID- 17907891 TI - [The antiphospholipid syndrome, an update]. AB - The antiphospholipid syndrome is an antibody mediated hypercoagulable state characterized by recurrent venous and arterial thromboembolic events. Several studies have determined that the frequency of antiphospholipid syndrome in patients presenting with a venous thromboembolic event is between 4% and 14%. Classical criteria include the presence of anticardiolipin antibody or lupus anticoagulant with typical complications of thrombosis or pregnancy loss. Other common associated manifestations include livedo reticularis, thrombocytopenia, valvular heart disease, and nephropathy with renal insufficiency, hypertension and proteinuria. Because of the high risk for recurrent thromboembolism in these patients, current recommendations suggest a longer, potentially lifelong, course of antithrombotic therapy following an initial event. For an initial venous thromboembolic event, a target INR of 2.0 to 3.0 is supported by two prospective, randomized clinical trials. In contrast, relatively limited data exist for an initial arterial thromboembolic event in patients who have the antiphospholipid syndrome, and therapeutic recommendations range from aspirin to warfarin with a high target INR. Recurrent thromboembolic events can be extremely difficult to treat, and some patients may benefit from the addition of immunosuppressive therapies. It is very important to evaluate in this setting additional, coincident prothrombotic risk factors. PMID- 17907892 TI - [The placebo in medical practice and research]. AB - The placebo is a therapeutics tool well known since the antiquity. It is used in clinical practice and research. In this article I have define the concept and resume the guidelines for its use in clinical research. PMID- 17907893 TI - [Mycoplasma pneumoniae pneumonia and adult respiratory distress syndrome]. PMID- 17907894 TI - [A new therapeutic strategy in breast cancer: metronomic chemotherapy]. PMID- 17907895 TI - [Diabetic foot and risk factors]. AB - OBJECTIVE: To identify clinic and metabolic risk factors for diabetic neuropathy and peripheral arterial disease in patients evaluated in a diabetic foot unit care. METHOD: From 2000 to 2005 we evaluated the presence of diabetic neuropathy (monofilament, tuning fork and Boulton's clinic scale) and peripheral arterial disease (ankle-brachial index and toe-brachial index) in 304 diabetic patients. We classified patients in four groups: patients without pathology (normal group), with neuropathy (neuropathic group), with peripheral arterial disease (vascular group) and with both pathologies (mixed group) and we compared the characteristics of each group. We analysed other poblational characteristics: age, gender, type of diabetes, duration, microvascular and macrovascular complications, hypertension, smoking habit, antiagregation and mean HbA1c in the last year. RESULTS: Age, frequency of hypertension and coronary disease were significantly higher (p < 0.005) in vascular and mixed group than in normal group (63 +/- 13 and 65 +/- 10 vs. 55 +/- 14; 69.2 and 70.3 vs. 45.5%; 46,2% and 39.2% vs 23.8%, respectively). Frequency of retinopathy, nephropathy and HbA1c were significantly higher (p < 0.05) in neuropathic and mix group than in normal group (62.5 and 66.2 vs. 32.7%; 45.3 and 47.3 vs. 24.8%; 8.1 +/- 1.6 and 8.0 +/- 1.3 vs 7.4 +/- 1.2 respectively). CONCLUSION: This study indicates that the development of diabetic neuropathy is related with worse metabolic control and the presence of other microvascular complications; while age, hypertension and coronary disease are risk factors for peripheral arterial disease. PMID- 17907896 TI - [Cross-sectional analysis of heart failure among patients in the Internal Medicine Service at a third-level hospital. Part I: epidemiologic analysis]. AB - OBJECTIVES: To observe the epidemiologic characteristics of the patients intake during five years in a internal medicine department, with heart failure. METHODS: A cross-sectional study of the intake patients in the Internal Medicine Service in the Hospital Clinico Universitario de Santiago de Compostela between 1999 to 2003. The variables analized were: sex, age, days of hospital stay, number of intake by failure cardiac, reason for admission (guide symptom), hypertension, diabetes mellitus, cardiac disease, fibrillation atrium, previous treatment with beta-blockers, blood pressure in the admission moment, to make echocardiography, disfunction systolic, etiology, deceased, treatment at the end. The statistical analysis was performed with qualitative and quantitative measures, chi-cuadrado and t-student, and multivariant analyses. RESULTS: 248 patients were accepted for the study. We observed more women than men (55.2%) and bigger median age (79 years old vs. 73 years old in men, p < 0.001). The mean income was 13.61 days and a median of 11 days. The 41,8% of the patients had hypertension, 30.9% diabetes mellitus and 81,9% had someone heart disease. The aetiologies of heart failure most frequent were ischemic cardiopathy (27.2%) and hypertension (24.2%). The most frequent symptom was the dyspnea (68.9%). It made echocardiography in 20.9% of patients and 45.1% showed systolic disfunction. The only factor related with this small percentage of echocardiographies was the incoming time. The most frequent etiology was respiratory infections (39.5%). The 8.6% of patients was deceased. The pharmacologic treatment more prescribed were the diuretics (86.9%) and transcutaneous nitrates (49.5%). It was indicated ECAI or AAR-II in the 86.9% of patients and beta-blockers in 0.9%. CONCLUSIONS: The number of echocardiograms practiced to the patients is smaller that the number advised by international associations and smaller to the cardiologist registers. The beta-blockers and ECAI use is smaller too. PMID- 17907897 TI - [Plasma concentrations of beta-endorphins in the children of alcoholic patients]. AB - BACKGROUND: Of the factors identified in different studies as the possible causes of alcoholism, heredity appears to be the most important. However, environmental factors can increase or decrease the risk of an individual developing alcohol dependence. METHOD: To clarify the possible influence of heredity on alcoholism, we studied the plasma concentration of beta-endorphins in 25 families with alcoholic members: 27 children whose father was alcoholic and 7 whose father and mother were both alcoholics. The results were compared with finding in an age matched control group of no-drinking adults and normal children in non-drinking families. RESULTS: The children of alcoholic parents had significantly lower beta endorphin levels (p < 0.001) than control individuals, and concentrations were especially low when both parents were alcoholics. CONCLUSION: We conclude that plasma beta-endorphin concentration may have predictive value in identifying persons likely to become alcoholics. PMID- 17907898 TI - [Is positron emission tomography (PET) scan useful for studying the extent of gastrointestinal stromal tumors]. AB - Gastrointestinal tumor (GIST) is the most frequent mesenchymal malignancy of the gastrointestinal tract. Liver and peritoneum are the most frequent metastatic sites. Surgery is the standard treatment in patients with localized disease and imatinib mesylate has been shown to be an effective treatment in metastatic or unresectable disease. We present a case of ileal GIST that was diagnosed as metastatic just with the findings of the fluorodeoxyglucose positron emission tomography (PET) and showed a total response with Imatinib on PET. PMID- 17907899 TI - [New diagnostic and surgical approach to Mirizzi syndrome]. AB - Mirizzi's syndrome is an unusual complication of gallstone disease, in which a stone impacting in the neck of the gallbladder (Hartmann s pouch) compresses the common bile duct. This mechanical obstruction leads to obstructive jaundice frequently followed by inflammatory changes and several complications. We present two patients affected by Mirizzi's syndrome whose diagnosis was correct in the preoperative period and approached by laparoscopy. A case was converted to open procedure due to adhesions in the Calot's triangle, and therefore, treated with subtotal cholecystectomy and choledochorrhaphy over a T tube. In the other case the laparoscopy access became successful. Both postoperative courses were uneventful. In this article, suitable diagnostic techniques are analyzed. On the other hand, we discuss what is the best therapeutic option, with a special attention to the relevance of endoscopic retrograde cholangiopancreatography and laparoscopic approach in the management of those patients. PMID- 17907900 TI - [Gastroesophageal reflux as a cause of night sweating]. AB - Night sweats has been defined as drenching sweats that require the patient to change bed clothes. In current studies night sweats appear in 30% of non obstetric patients and affects approximately 60% of pregnant women. Differential diagnoses include infections, malignancy, medications, hot flashes and panic attacks, making of each patient a challenge. We present two patients with night sweating. After excluding systemic diseases the diagnosis of gastroesophageal reflux was made, with excellent response to anti-reflux treatment. The presentation of our two patients coupled with a deep literature review, underscores the importance of gastroesophageal reflux as a cause of night sweating. PMID- 17907901 TI - [Lung adenocarcinoma feigning an interstitial lung disease in a 30-year-old man]. AB - A case of a 30-year-old man is presented. He had a past medical history of asthma and presented with cough, shortness of breath, fever and chest pain. The chest X ray showed cardiomegaly and a interstitial pattern. The echocardiogram revealed a severe pericardial effusion that required performing a pericardiocentesis. A CT scan showed mediastinal adenopathies and reticulonodular interstitial pattern. A bronchoscopy could not be completed because of non-tolerance. Finally a lung biopsy allowed us to reach a final diagnosis of lung adenocarcinoma. Following, some aspects of lung cancer in young people are commented, especially its incidence, histology, symptoms and a prognosis. A few aspects of the interstitial lung disease are also addressed and the differential diagnosis with lymphangitic carcinomatosis. PMID- 17907902 TI - [Chlamydial infections]. AB - Chlamydiae are a singular group of bacteria, with a vital cycle that is intracellular in part. These microorganisms adhere to epithelia, where they may provoke infection. There are three pathogenic species for humans: Chlamydia trachomatis, which produce infections mainly in the genital and urinary tracts, and C. psittaci and C. pneumoniae, which mostly produce respiratory infections. The last two microorganisms are now classified in a different genus, known as Chlamydophila. PMID- 17907903 TI - [Machado, Moix and Montalban: smoke gets in your lives]. AB - The writers Antonio Machado (pulmonary chronic disease), Terenci Moix (emphysema) and Manuel Vazquez Montalban (acute myocardial infarction) died of the complications derived from their inveterate habit-forming tobacco. They were aware of the prejudices that a such addiction was causing and tried, in some moment, to leave it. In addition, the testimonies of their contemporary or their same writings offered dramatic samples of this struggle. Nevertheless, they relapsed and tobacco caused them a premature death before 65 years. With a panoramic of 60 years among the last two authors and Machado, a brief historical perspective is offered on the acquisition of scientific knowledge that changed the attitudes towards tobacco during this period of time. PMID- 17907904 TI - [Postpartum thyroiditis with unusual evolution]. PMID- 17907905 TI - Buried energy reclaimed: running a locomotive and other delayed fulfillments. PMID- 17907906 TI - An inquiry into the sources of poetic vision: Part I -- the path to inspiration. AB - An inquiry is made into the peculiar way artists come to know things. It is agreed that artists have an unusual access to unconscious material, but how they gain such access and what else informs their vision is further explored. Two conditions are found necessary to the artist's access: a limited awareness of his or her meaning and an altered state of consciousness brought on by a special form of play. The essential features of play are examined, including its social dimensions. The artist's vision is shown to arise from a free play between his own mind, the material he works in, and his audience. The artist's mind is informed by ideas and techniques available to her (her position in the history of ideas) and by her own unconscious concerns. The artist's vision is transformed by the play between his mind (conscious and unconscious), his subject, and his audience. PMID- 17907908 TI - A view from Riggs: treatment resistance and patient authority -- V. Silencing the messenger: the social dynamics of treatment resistance. AB - Treatment resistance has emerged as a significant issue in our era emphasizing biological treatments for psychiatric disorders. Some treatment programs utilize milieu treatment to advance work with treatment resistant patients. This article (1) explores the social factors that contribute to and exacerbate treatment resistance (2) reviews the theoretical framework and rationale of a therapeutic community milieu treatment program, and (3) illustrates therapeutic community work with two treatment resistant patients. PMID- 17907909 TI - Turning passive into active: a building block of ego and fundamental mechanism of defense. AB - Turning passive into active is an ego function that plays an important role in managing anxiety associated with passive feelings of helplessness and powerlessness. Its use in childhood aids the ego in constructing a basic sense of mastery, a secure sense of being in control, both of oneself and of one's circumstances. Functioning first as a building block of ego, it subsequently becomes a versatile mechanism of defense with utility throughout the life cycle. This article discusses both the developmental and defensive uses of the passive into active mechanism of defense and illustrates them with clinical examples. Often regarded only as a byproduct of defense, its importance in mastering core developmental anxieties and their adult revivals warrants its inclusion in the glossary of discrete defenses. PMID- 17907910 TI - Amplification of the concept of erroneous meaning in psychodynamic science and in the consulting room. AB - Previous papers dealt with the concept of psyche as that dynamic field which underlies the subjective experience of mind. A new paradigm, psychodynamic science, was suggested for dealing with subjective data. The venue of the psychotherapeutic consulting room is now brought directly into science, expanding the definition of psychotherapy to include both humanistic and scientific elements. Certain concepts were introduced to amplify this new scientific model, including psyche as hypothetical construct, the concept of meaning as replacement for operational validation in scientific investigation, the synonymity of meaning and insight, and the concept of synchronicity, together with the meaning connected affect of numinosity. The presence of unhealthy anxiety as the conservative ego attempts to preserve its integrity requires a deeper look at the concept of meaning. This leads to a distinction between meaning and erroneous meaning. The main body of this paper amplifies that distinction, and introduces the concept of intolerance of ambiguity in the understanding of erroneous meanings and their connection with human neurosis. PMID- 17907911 TI - Psychodynamic psychiatry in community settings. AB - Psychodynamics are surprisingly contemporary for community-based psychiatry. Clinical vignettes illustrate this point. Familiarity with literature concerning symbolic communication, transference, projective identification, and narcissism help provide perspective. Psychiatry in community settings can be stressful. Its practice is improved by supervised experience, grounded in psychodynamics. PMID- 17907912 TI - The timing of psychodynamically oriented psychotherapy termination and its relation to reasons for termination, feelings about termination, and satisfaction with therapy. AB - The present study investigated retrospective client accounts of the timing of psychodynamically oriented private practice psychotherapy termination and its relation to the reasons for termination on the one hand, and to client feelings about termination and satisfaction from therapy on the other. Eighty-two persons who had been in private practice psychodynamically oriented psychotherapy were assessed using open-ended questions and rating scales. Forty percent of participants reported that therapy ended on time, 37% that it ended earlier than it should have, and 23% that it ended later than it should have. In general, clients who reported that termination was on time were more satisfied, terminated for less negative reasons, and had more positive feelings. Results suggest that clients find terminating psychotherapy at the right time important and yet difficult to achieve. PMID- 17907913 TI - Money and sentiment: a psychodynamic approach to behavioral finance. AB - This article tackles one of the timeliest issues for both practitioners and patients today: sentiment, psychodynamics, and the stock market. Economic bubbles and crashes have occurred regularly through history -- from Holland's 17th century tulip mania, to America's 19th century railway mania, to the 1990s high tech obsession. Though most investors regard themselves as investing rationally, few do. Instead they react collectively, buying high and selling low in crowds. Being subject to the illusion of control, they follow regressive behavior patterns and irrational, wishful thinking. They are victimized by their own emotions of hope, fear, and uncertainty. Crises happen often in economics. Indeed, the market itself may be quantified as a conglomeration of human sentiment. The relationship between magical thinking and the pictorial language of the market will be explored. Psychodynamic conceptualizations about risk and speculation are discussed, as are the interplay of affects versus judgment, rational thinking, and the knowledge of one's own capacity for stress tolerance. PMID- 17907914 TI - Introduction to "communicating with the schizophrenic superego revisited: a new technique". PMID- 17907915 TI - Communicating with the schizophrenic superego revisited: a new technique. AB - Developing from the author's prior work in which the patient's superego is conceived as more accessible to consciousness than generally believed, this article undertakes greater illumination of technical matters arising in the clinical circumstance. Directly and firmly addressing the superego of the psychotic patient is regarded as having immediate therapeutic benefit, rendering the ego stronger and leading to increased rationality and firmer sense of identity. PMID- 17907917 TI - Assessing population risk for postmenopausal osteoporosis: a new strategy using data from the Behavioral Risk Factor Surveillance System (BRFSS). AB - Osteoporosis public health measures are hindered by the inability to easily identify subclinical disease. We have now estimated state-specific osteoporosis prevalences using a simple formula (OST Index) to analyze age and weight of 62,882 older women; the prevalences determined are similar to those based on BMD. This new method has potential use for guiding implementation of osteoporosis prevention/treatment programs. INTRODUCTION: Although osteoporosis-related fractures are a major U.S. public health issue, population-based prevention programs have not yet been developed. One contributing factor has been lack of a suitable screening test to detect asymptomatic high-risk individuals. MATERIALS AND METHODS: We estimated state-specific prevalences of postmenopausal osteoporosis using the Osteoporosis Self-Assessment Tool Index (OST Index; [self reported weight in kg - age] x 0.2) to analyze data from 62,882 women >or=50 yr of age who participated in the 2002 Behavioral Risk Factor Surveillance System (BRFSS). The OST Index, designed to assess an individual's risk of disease, has previously been shown to have modest positive and high negative predictive value for osteoporosis defined by BMD criteria. Based on this index, women from each state were distributed among high-, moderate-, and low-risk OST categories. Calculated percentages for each category were weighted to U.S. Census Bureau population projections for 2002. By adjusting results to reflect previously validated percentages of women with osteoporosis in each risk category, we estimated the prevalence of postmenopausal osteoporosis in each state. RESULTS: Our calculated weighted prevalence estimates agreed closely with those of the National Osteoporosis Foundation derived from actual femoral neck BMD measurements obtained in the third National Health and Nutrition Examination Survey (1988-1994) and projected to U.S. census state population predictions for 2002. Comparison of unweighted BRFSS-OST results and NHANES BMD data revealed similar percentages of osteoporosis among all women >or=50 yr of age (BRFSS, 18.5%; NHANES, 18.0%; p = 0.47) and also among white women (BRFSS, 19.0%; NHANES, 20.0%; p = 0.28). However, the percentages of osteoporosis among blacks and Hispanics did not correspond, at least partly because of the lack of race specific reference standards for BMD measurements and OST index ranges. CONCLUSIONS: Analysis of readily available BRFSS data with the OST index formula is a simple, no-cost technique that provides state prevalence estimates of postmenopausal osteoporosis that could be used to guide allocation of resources to statewide osteoporosis prevention programs. PMID- 17907918 TI - Osteoblast-targeted expression of Sfrp4 in mice results in low bone mass. AB - Transgenic mice overexpressing Sfrp4 in osteoblasts were established. These mice exhibited low bone mass caused by a decrease in bone formation. INTRODUCTION: We recently reported that single nucleotide polymorphisms in the secreted frizzled related protein 4 (Sfrp4) gene are responsible for low peak BMD in senescence accelerated mouse (SAM) P6. In vitro studies revealed inhibition of osteoblast proliferation by Sfrp4, which is supposed to be mediated by canonical Wnt signaling. MATERIALS AND METHODS: We examined the expression of Sfrp4 in neonate long bones by in situ hybridization and generated transgenic mice in which Sfrp4 was specifically overexpressed in osteoblasts under the control of a 2.3-kb Col1a1 osteoblast-specific promoter. Next, we compared the phenotype of Sfrp4 transgenic (Sfrp4 TG) mice with that of mice in which one allele of beta-catenin was conditionally disrupted in osteoblasts (betaChet), and administered lithium chloride (LiCl) to Sfrp4 TG mice. RESULTS: Hemizygous Sfrp4 TG mice exhibited a 30% reduction of trabecular bone mass compared with that in wildtype littermates at 8 wk of age, and histomorphometrical analysis showed decreases in both osteoblast numbers and bone formation rate. betaChet mice exhibited a 17% reduction of trabecular bone mass in distal femora caused by an increase in the osteoclast number and a decrease in bone formation rate. Furthermore, LiCl administration rescued the bone phenotype of Sfrp4 TG mice. CONCLUSIONS: Expression of Sfrp4 in periosteum and bone tissues suggested the role of Sfrp4 in osteoblasts, and we identified that overexpression of Sfrp4 in osteoblasts suppressed osteoblast proliferation, resulting in a decrease in bone formation in vivo. Partial suppression of beta-catenin/canonical Wnt signaling also impaired bone formation, and activation of the signaling restored low bone mass of Sfrp4 TG mice. Thus, these results indicate that Sfrp4 decreases bone formation at least in part by attenuating canonical Wnt signaling in vivo. PMID- 17907919 TI - Epistatic effects contribute to variation in BMD in Fischer 344 x Lewis F2 rats. AB - To further delineate the factors underlying the complex genetic architecture of BMD in the rat model, a genome screen for epistatic interactions was conducted. Several significant interactions were identified, involving both previously identified and novel QTLs. INTRODUCTION: The variation in several of the risk factors for osteoporotic fracture, including BMD, has been shown to be caused largely by genetic differences. However, the genetic architecture of BMD is complex in both humans and in model organisms. We have previously reported quantitative trait locus (QTL) results for BMD from a genome screen of 595 female F(2) progeny of Fischer 344 and Lewis rats. These progeny also provide an excellent opportunity to search for epistatic effects, or interaction between genetic loci, that contribute to fracture risk. MATERIALS AND METHODS: Microsatellite marker data from a 20-cM genome screen was analyzed along with weight-adjusted BMD (DXA and pQCT) phenotypic data using the R/qtl software package. Genotype and phenotype data were permuted to determine a genome-wide significance threshold for the epistasis or interaction LOD score corresponding to an alpha level of 0.01. RESULTS AND CONCLUSIONS: Novel loci on chromosomes 12 and 15 showed a strong epistatic effect on total BMD at the femoral midshaft by pQCT (LOD = 5.4). A previously reported QTL on chromosome 7 was found to interact with a novel locus on chromosome 20 to affect whole lumbar BMD by pQCT (LOD = 6.2). These results provide new information regarding the mode of action of previously identified rat QTLs, as well as identifying novel loci that act in combination with known QTLs or with other novel loci to contribute to the risk factors for osteoporotic fracture. PMID- 17907920 TI - Serum 25-hydroxyvitamin D and hip fracture risk in older U.S. white adults. AB - We used serum 25(OH)D data from NHANES III and incident hip fracture cases identified using linked mortality and Medicare records, and found that serum 25(OH)D was significantly related to reduced hip fracture risk in non-Hispanic white adults >or=65 yr of age. INTRODUCTION: The role of vitamin D status in reducing fracture risk is unclear. We examined the relationship between serum 25 hydroxyvitamin D [25(OH)D] and incident hip fracture risk in older non-Hispanic white adults. MATERIALS AND METHODS: The study sample consisted of 1917 white men and women >or=65 yr of age who were examined in the third National Health and Nutrition Examination Survey (NHANES III, 1988-1994), a nationally representative survey. Incident hip fractures were ascertained using linked mortality and Medicare records that were obtained for NHANES III participants. Serum 25(OH)D values were measured with a radioimmunoassay kit. Cox proportional hazards models were used to estimate the relative risk (RR) of hip fracture by serum 25(OH)D level. RESULTS: There were 156 incident hip fracture cases in the sample. Cases were older, had lower BMD and body mass index, more prevalent spine or wrist fractures and weight loss before baseline, and ate fewer kilocalories and less calcium than noncases. After adjusting for these differences, serum 25(OH)D values exceeding 60 nM were significantly related to hip fracture risk. For example, the multivariate-adjusted RR was 0.64 (95% CI, 0.46-0.89) among individuals with serum 25(OH)D values >or=62.5 nM compared with those with values below this level. When grouped into quartiles, the multivariate-adjusted RR for the second, third, and fourth versus the first quartile of serum 25(OH)D were 0.50 (95% CI, 0.25-1.00), 0.41 (95% CI, 0.24-0.70), and 0.50 (95% CI, 0.29-0.86), respectively. CONCLUSIONS: Serum 25(OH)D was related to a significantly lower hip fracture risk in this cohort of older white adults, even after adjusting for several relevant confounding variables. The relationship did not seem to be linear across all values. Our results support other studies suggesting that serum 25(OH)D values exceeding 60 nM are associated with health benefits. PMID- 17907922 TI - Identification of a major locus for Paget's disease on chromosome 10p13 in families of British descent. AB - Mutations of SQSTM1 are an important cause of PDB, but other genes remain to be discovered. A major susceptibility locus for PDB was identified on chromosome 10p13 by a genome-wide linkage scan in families of British descent, which accounted for the vast majority of cases not caused by SQSTM1 mutations. INTRODUCTION: Paget's disease of bone (PDB) has a strong genetic component, and several susceptibility loci have been identified by genome-wide linkage scans. We previously identified three susceptibility loci for PDB using this approach on chromosomes 5q35, 2q36, and 10p13 in 62 families of mainly British descent, but subsequently, mutations in the SQSTM1 gene were found to be the cause of PDB in 23 families from this cohort. Here we reanalyzed the results of our genome-wide search in families from this cohort who did not have SQSTM1 mutations. MATERIALS AND METHODS: The study population consisted of 210 individuals from 39 families of predominantly British descent with autosomal dominant inheritance of PDB in whom SQSTM1 mutations had been excluded by mutation screening. The average family size was 5.44 +/- 3.98 (SD) individuals (range, 2-24 individuals). Genotyping was performed using standard techniques with 382 microsatellite markers spaced at an average distance of 9.06 cM throughout the autosomes. Multipoint linkage analysis was performed using the GENEHUNTER program under models of homogeneity and heterogeneity. RESULTS: Multipoint parametric linkage analysis under a model of homogeneity and nonparametric linkage analysis under a model of heterogeneity both showed strong evidence of linkage to a single locus on chromosome 10p13 (LOD score, +4.08) close to the marker D10S1653 at 41.43cM. No evidence of linkage was detected at the chromosome 2q36 locus previously identified in this population, and linkage to other candidate loci previously implicated in the pathogenesis of PDB was excluded. CONCLUSIONS: We conclude that there is an important susceptibility gene for PDB on chromosome 10p13 in families of British descent and find no evidence to support the existence of a susceptibility locus on chromosome 2q36 or other previously identified candidate loci for PDB in this population. The gene that lies within the 10p13 locus seems to account for the development of PDB in the vast majority of families of British descent who do not carry SQSTM1 mutations. PMID- 17907921 TI - Complete volumetric decomposition of individual trabecular plates and rods and its morphological correlations with anisotropic elastic moduli in human trabecular bone. AB - Trabecular plates and rods are important microarchitectural features in determining mechanical properties of trabecular bone. A complete volumetric decomposition of individual trabecular plates and rods was used to assess the orientation and morphology of 71 human trabecular bone samples. The ITS-based morphological analyses better characterize microarchitecture and help predict anisotropic mechanical properties of trabecular bone. INTRODUCTION: Standard morphological analyses of trabecular architecture lack explicit segmentations of individual trabecular plates and rods. In this study, a complete volumetric decomposition technique was developed to segment trabecular bone microstructure into individual plates and rods. Contributions of trabecular type-associated morphological parameters to the anisotropic elastic moduli of trabecular bone were studied. MATERIALS AND METHODS: Seventy-one human trabecular bone samples from the femoral neck (FN), tibia, and vertebral body (VB) were imaged using muCT or serial milling. Complete volumetric decomposition was applied to segment trabecular bone microstructure into individual plates and rods. The orientation of each individual trabecula was determined, and the axial bone volume fractions (aBV/TV), axially aligned bone volume fraction along each orthotropic axis, were correlated with the elastic moduli. The microstructural type-associated morphological parameters were derived and compared with standard morphological parameters. Their contributions to the anisotropic elastic moduli, calculated by finite element analysis (FEA), were evaluated and compared. RESULTS: The distribution of trabecular orientation suggested that longitudinal plates and transverse rods dominate at all three anatomic sites. aBV/TV along each axis, in general, showed a better correlation with the axial elastic modulus (r(2) = 0.95 approximately 0.99) compared with BV/TV (r(2) = 0.93 approximately 0.94). The plate-associated morphological parameters generally showed higher correlations with the corresponding standard morphological parameters than the rod-associated parameters. Multiple linear regression models of six elastic moduli with individual trabeculae segmentation (ITS)-based morphological parameters (adjusted r(2) = 0.95 approximately 0.98) performed equally well as those with standard morphological parameters (adjusted r(2) = 0.94 approximately 0.97) but revealed specific contributions from individual trabecular plates or rods. CONCLUSIONS: The ITS-based morphological analyses provide a better characterization of the morphology and trabecular orientation of trabecular bone. The axial loading of trabecular bone is mainly sustained by the axially aligned trabecular bone volume. Results suggest that trabecular plates dominate the overall elastic properties of trabecular bone. PMID- 17907923 TI - Growth and bone mineral accretion during puberty in Chinese girls: a five-year longitudinal study. AB - There are few longitudinal data on bone development during puberty in children with low calcium intake. This 5-yr longitudinal study showed that, in Chinese girls, the mean apparent calcium retention efficiency during puberty was 40.9%, PHV occurred at 3-0 yr before menarche, and peak bone mineral accretion occurred 1 yr later than PHV. Chinese girls have high calcium retention efficiency during puberty. INTRODUCTION: There are few longitudinal data on bone development during puberty in children with low dietary calcium intake. The aim of this study was to examine the rate of growth and bone mineral accretion and study the predictors of total body BMC during puberty in a 5-yr longitudinal study with Chinese girls. MATERIALS AND METHODS: Ninety-two girls, 9.5-10.5 yr of age at baseline, from the unsupplemented control group of a school milk intervention trial were included in this analysis. Data on anthropometric measurements, total body BMC as assessed by DXA, and calcium intake as assessed by a 3-day food record were obtained at baseline and 1, 2, 4, and 5 yr. RESULTS: The mean age of menarche was 12.1 +/- 1.0 yr. The mean annual rate of bone mineral accretion was 197.4 g/yr during the follow-up period, representing a calcium accretion rate of 162.3 mg/d. This calcium retention rate and the average dietary calcium intake of 444.1 mg/d gave an apparent calcium retention efficiency of 40.9%. Peak height velocity (PHV) occurred at 3-0 yr before menarche. Peak bone mineral accretion occurred 1 yr later than PHV. There was a decrease in size-corrected BMD in the year before menarche. In the linear mixed-effects model analysis containing body size and lifestyle factors, we found that height, body weight, and calcium intake were significant independent predictors of total body BMC. CONCLUSIONS: Chinese girls with low habitual dietary calcium intake have high calcium retention efficiency during puberty. Because calcium intake is a significant predictor of total body BMC, increasing dietary calcium intake may have beneficial effects on bone mineral accretion in these girls. PMID- 17907924 TI - Interaction of galectin-9 with lipid rafts induces osteoblast proliferation through the c-Src/ERK signaling pathway. AB - Galectin-9 is a beta-galactoside-binding lectin expressed in various tissues, including bone. The role of galectin-9 in human osteoblasts, however, remains unclear. This study showed that galectin-9 interacts with lipid rafts and induces osteoblast proliferation through the c-Src/ERK signaling pathway. INTRODUCTION: Galectin-9 is a beta-galactoside-binding lectin that modulates many biological functions by interacting with particular carbohydrates attached to proteins and lipids. However, the role of galectin-9 in bone metabolism and osteoblast proliferation remains unclear. This study investigated the effects of galectin-9 on osteoblast proliferation and its signaling mechanisms. MATERIALS AND METHODS: The effect of galectin-9 on osteoblast proliferation was tested by measuring the conversion of tetrazolium salt WST-8 to formazan. Protein phosphorylation was assayed by western blotting and confocal microscopy was used to localize lipid rafts. RESULTS: Galectin-9-induced proliferation of the obtained osteoblasts in a dose-dependent manner, whereas galectin-1, -3, and -4 did not. Galectin-9-induced phosphorylation of c-Src and subsequent ERK1/ERK2 in the osteoblasts. The galectin-9-induced phosphorylation and proliferation were inhibited by PP2, a selective inhibitor of c-Src. Galectin-9-induced clustering of lipid rafts detected by cholera toxin B (CTB; binding the raft-resident ganglioside GM1) using confocal microscopy. Cross-linking of the GM1 ganglioside with CTB by anti CTB antibody-induced phosphorylation of c-Src, whereas disruption of galectin-9 induced lipid rafts by beta-methylcyclodextrin reduced c-Src phosphorylation and proliferation of the cells. CONCLUSIONS: These results suggest that galectin-9, but not other galectins, induced proliferation of human osteoblasts through clustering lipid rafts on membrane and subsequent phosphorylation of the c Src/ERK signaling pathway. PMID- 17907926 TI - Current prospects for the generation of patient-specific pluripotent cells from adult tissues. PMID- 17907925 TI - Osteoblasts express NLRP3, a nucleotide-binding domain and leucine-rich repeat region containing receptor implicated in bacterially induced cell death. AB - Bacterially induced osteoblast apoptosis may be a major contributor to bone loss during osteomyelitis. We provide evidence for the functional expression in osteoblasts of NLRP3, a member of the NLR family of cytosolic receptors that has been implicated in the initiation of programmed cell death. INTRODUCTION: Osteoblasts undergo apoptosis after exposure to intracellular bacterial pathogens commonly associated with osteomyelitis. Death of this bone-forming cell type, in conjunction with increased numbers and activity of osteoclasts, may underlie the destruction of bone tissue at sites of bacterial infection. To date, the mechanisms responsible for bacterially induced apoptotic osteoblast cell death have not been resolved. MATERIALS AND METHODS: We used flow cytometric techniques to determine whether intracellular invasion is needed for maximal apoptotic cell death in primary osteoblasts after challenge with Salmonella enterica. In addition, we used real-time PCR and immunoblot analyses to assess osteoblast expression of members of the nucleotide-binding domain leucine-rich repeat region containing family of intracellular receptors (NLRs) that have been predicted to be involved in the induction of programmed cell death. Furthermore, we have used co-immunoprecipitation and siRNA techniques to confirm the functionality of such sensors in this cell type. RESULTS: In this study, we showed that invasion of osteoblasts by Salmonella is necessary for maximal induction of apoptosis. We showed that murine and human osteoblasts express NLRP3 (previously known as CIAS1, cryopyrin, PYPAF1, or NALP3) but not NLRC4 (IPAF) and showed that the level of expression of this cytosolic receptor is modulated after bacterial challenge. We showed that osteoblasts express ASC, an adaptor molecule for NLRP3, and that these molecules associate after Salmonella infection. In addition, we showed that a reduction in the expression of NLRP3 attenuates Salmonella-induced reductions in the activity of an anti-apoptotic transcription factor in osteoblasts. Furthermore, we showed that NLRP3 expression is needed for caspase-1 activation and maximal induction of apoptosis in osteoblasts after infection with Salmonella. CONCLUSIONS: The functional expression of NLRP3 in osteoblasts provides a potential mechanism underlying apoptotic cell death of this cell type after challenge with intracellular bacterial pathogens and may be a significant contributory factor to bone loss at sites of infection. PMID- 17907927 TI - Lessons for the nascent regenerative medicine industry from the biotech sector. PMID- 17907930 TI - Therapeutic neutralization of CXCL10 decreases secondary degeneration and functional deficit after spinal cord injury in mice. AB - Inflammation plays a critical role in the secondary degenerative response to spinal cord injury (SCI). The influx of inflammatory cells following SCI is preceded by the expression of specific chemoattractants, including chemokines. The chemokine CXCL10 is a potent T lymphocyte recruiter and has been strongly implicated in the pathology of many CNS disorders. We have previously demonstrated that CXCL10 exacerbates secondary degeneration by blocking the function of CXCL10 prior to SCI. Here we administered neutralizing antibodies against CXCL10 1 h after SCI in order to investigate the efficacy of this therapeutic intervention in abating histologic and functional deficit following acute SCI and further assess the functional role of CXCL10 in secondary degeneration. Neutralization of CXCL10 significantly reduced inflammation, apoptosis, neuronal loss and whole tissue loss. Notably, this therapeutic treatment also promoted revascularization of the injured spinal cord and functional recovery. These data suggest that anti-CXCL10 antibody treatment is a viable therapeutic strategy for acute SCI. PMID- 17907931 TI - New approach to radiation burn treatment by dosimetry-guided surgery combined with autologous mesenchymal stem cell therapy. AB - The therapeutic management of severe radiation burns remains a challenging issue. Conventional surgical treatment (excision and skin autograft or rotation flap) often fails to prevent unpredictable and uncontrolled extension of the radiation necrotic process. We report here an innovative therapeutic strategy applied to the victim of a radiation accident (December 15, 2005) with an iridium gammagraphy radioactive source (192Ir, 3.3 TBq). The approach combined numerical dosimetry-guided surgery with cellular therapy using mesenchymal stem cells. A very severe buttock radiation burn (2000 Gy at the center of the skin surface lesion) of a 27-year-old Chilean victim was widely excised (10 cm in diameter) using a physical and anatomical dose reconstruction in order to better define the limit of the surgical excision in apparently healthy tissues. A secondary extension of the radiation necrosis led to a new excision of fibronecrotic tissues associated with a local cellular therapy using autologous expanded mesenchymal stem cells as a source of trophic factors to promote tissue regeneration. Bone marrow-derived mesenchymal stem cells were expanded according to a clinical-grade technique using closed culture devices and serum-free medium enriched in human platelet lysate. The clinical evolution (radiation pain and healing progression) was favorable and no recurrence of radiation inflammatory waves was observed during the 11 month patient's follow-up. This novel multidisciplinary therapeutic approach combining physical techniques, surgical procedures and cellular therapy with adult stem cells may be of clinical relevance for improving the medical management of severe localized irradiations. It may open new prospects in the field of radiotherapy complications. PMID- 17907932 TI - Epigenetic dedifferentiation of somatic cells into pluripotency: cellular alchemy in the age of regenerative medicine? AB - Ever since the derivation of the first human embryonic stem cell line, hopes have persisted for the treatment of a wide range of cellular degenerative diseases. However, significant immuno-incompatibility between donor cells and recipient patients remains an unsolved challenge. Currently, three main strategies are investigated in humans to create autologous pluripotent stem cells: somatic cell nuclear transfer, cell fusion and cell extract incubation. All methods exploit the fact that a somatic genome is amenable to epigenetic dedifferentiation into a more plastic state, presumably through direct exposure to and manipulation by heterologous transcriptional factors. Epigenetic reprogramming includes profound modifications of chromatin structure, but the responsible mechanisms that work in toti- and pluripotent cells remain largely unknown. This review presents a brief introduction to stem cell terminology and epigenetics, followed by a critical examination of the predominant methodologies involved. Finally, the search for specific reprogramming factors is discussed, and obstacles for the clinical implementation of reprogrammed cells are addressed. PMID- 17907933 TI - Cellular approaches for stimulating CNS remyelination. AB - Myelination is critical for the normal functioning of the vertebrate nervous system. In the CNS, myelin is produced by oligodendrocytes, and the loss of oligodendrocytes and myelin results in severe functional impairment. Although spontaneous remyelination occurs in chronic demyelinating diseases such as multiple sclerosis, the repair process eventually fails, often resulting in long term disability. Two distinct general approaches can be considered to promote myelin repair. In one the target is stimulation of the endogenous myelin repair process through delivery of growth factors, and in the second the target is augmentation of the repair process through the delivery of exogenous cells with myelination potential. In both cases, effective treatment of diseases such as multiple sclerosis requires modulation of the immune system, since demyelination is associated with specific immunological activation. Recent studies have shown that some populations of stem cells, including mesenchymal stem cells, have the capacity of promoting endogenous myelin repair and modulating the immune response, prompting an assessment of their use as therapy in demyelinating diseases such as MS. Other types of demyelinating disorders, such as the leukodystrophies, may require multiple repair strategies including both replacement of dysfunctional cells and delivery or supplementation of growth factors, immune modulators or metabolic enzymes. Here we discuss the use of stem cells for the treatment of demyelinating diseases. While the current number of stem cell-based clinical trials for demyelinating diseases is limited, this is likely to increase significantly in the next few years, and a clear understanding of the applicability, limitations and underlying mechanisms mediating stem cell repair is critical. PMID- 17907934 TI - Vascular tissue engineering and vascularized 3D tissue regeneration. AB - Vascularized tissue regeneration has a great deal of potential in clinical medicine. Appropriate 3D tissue regeneration that yields tissue with the desired function and shape requires both growth signals and vascularization. In this paper, we discuss vascularized tissue regeneration using various vessel systems: artificial vessel, autologous vascular graft, autologous vascular bundle transfer and tissue engineered vessel. Vascularized 3D tissue regeneration will require a great deal of additional research before it can be applied to clinical situations. Several promising studies of vascularized tissue regeneration have been reported. However, additional studies into the maturation of neovascularization, the development of effective biomaterial, and the possibility of using stem cells will be needed before these techniques can be used in the clinical situation. PMID- 17907935 TI - Will regulation determine the science agenda? A look at hESCs. AB - Given the significant controversy over human embryonic stem cell (hESC) isolation and research, regulation of such work around the world has proceeded in an uncoordinated manner. In general, advances in science cause a need or desire for regulation; however, it has been the opposite for hESC research--regulation and policy have set certain boundaries for scientific research and defined other research questions. This is especially evident in the USA, where federal funding policies have engendered specific research towards novel methods for isolating such cells that do not require destruction of human embryos. Due to the multiplicity of national policies, it will be almost impossible to reach global consensus in the near future. Nonetheless, this paradigm of regulation leading science may have significant implications for future research projects. Changes in hESC policy in the short term will influence longer-term research potential. PMID- 17907936 TI - Stopping diabetes in its tracks: autologous non-myeloablative stem cell transplantation. AB - A recent report in this year's April issue of Journal of the American Medical Association describes an unprecedented success in delaying insulin dependence in patients with recent-onset Type 1 diabetes after non-myeloablative immune suppression with cyclophosphamide and antithymocyte globulin followed by autologous stem cell transplantation. In this study, 14 out of 15 patients became insulin-independent, which lasted up to 35 months. Concomitantly, C-peptide levels increased substantially compared with preintervention values. Treatment of autoimmune disorders, and in particular Type 1 diabetes, constitutes a complex balancing act between suppressing autoaggressive responses strongly and permanently enough, while circumventing much-feared long-term side effects from chronic immunosuppression. This clinical Phase I/II trial is relevant to fine tuning interventive protocols and contributing to the further development of suitable combination therapies to prevent and treat Type 1 diabetes. PMID- 17907937 TI - Obituary: Dame Dr Anne McLaren. PMID- 17907939 TI - Embryo stem cells from parthenotes and embryos produced by nuclear transfer: the distinction between them and their potential value in cell therapy. PMID- 17907940 TI - DNA methylation errors in cloned mice disappear with advancement of aging. AB - Cloned animals have various health problems. Aberrant DNA methylation is a possible cause of the problems. Restriction landmark genomic scanning (RLGS) that enabled us to analyze more than 1,000 CpG islands simultaneously demonstrated that all cloned newborns had aberrant DNA methylation. To study whether this aberration persists throughout the life of cloned individuals, we examined genome wide DNA methylation status of newborn (19.5 dpc, n=2), adult (8-11 months old, n=3), and aged (23-27 months old, n=4) cloned mice using kidney cells as representatives. In the adult and aged groups, cloning was repeated using cumulus cells of the adult founder clone of each group as nucleus donor. Two newborn clones had three with aberrantly methylated loci, which is consistent with previous reports that all cloned newborns had DNA methylation aberrations. Interestingly, we could detect only one aberrantly methylated locus in two of the three adult clones in mid-age and none of four senescent clones, indicating that errors in DNA methylation disappear with advancement of animals' aging. PMID- 17907941 TI - Overcoming MIII arrest from spontaneous activation in cultured rat oocytes. AB - The rat oocyte spontaneously activates under a wide variety of conditions. This process progresses to MIII arrest that is not responsive to parthenogenetic activation and development. Insofar as activation involves extrusion of the second polar body (PBII), we set out to determine if preventing this step by inhibiting microfilaments would change the course of spontaneous activation (SA). In particular, how long does the effect of SA persist while retaining reversibility of PBII extrusion once inhibitors are removed? We wanted to determine if the eggs would be responsive to parthenogenetic activation and capable of resuming development once a permanent inhibition is achieved. We set out to determine whether SA would depend on the ovular age of oocytes. Inhibiting of PBII extrusion was achieved by affecting microtubules with demecolcine or nocodazole or actin filaments with cytochalasin B (CB) and cytochalasin D (CD). We found that all oocytes undergo SA and progression to MIII; however, the rapidity of spontaneous activation is a function of the ovular age of the oocyte. The resumption of the meiosis period changes dramatically from 20 to 180 min with decreasing ovular age. We established that suppression of PB formation can be effectively achieved in oocytes of younger ovular age, and that inhibition of PB extrusion became irreversible after 3.5 h of treatment. We established that drug treated oocytes could undergo subsequent reactivation and in vitro development to blastocysts. The rate of in vitro development of cytochalasin-treated group was comparable to parthenogenetic controls, while nocodazole and demecolcine produced oocytes that developed at lower frequencies. Thus, the application of the microfilament inhibiting drugs helps to overcome the negative effect of SA that results in MIII arrest. Here we also show optimized parthenogenetic stimulation that resulted in development to the blastocyst stage at frequency comparable to development of fertilized embryos. PMID- 17907942 TI - EGF and TGF-alpha supplementation enhances development of cloned mouse embryos. AB - In this study, we sought to determine the extent to which mitogenic growth factors affect the survival and development of cloned mouse embryos in vitro. Cloned embryos derived by intracytoplasmic nuclear injection (ICNI) of cumulus cell nuclei into enucleated oocytes were incubated in culture media supplemented with EGF and/or TGF-alpha for 4 days. Compared to control, treatment with either growth factor significantly increased the blastocyst formation rate, the total number of cells per blastocyst, the cell ratio of the inner cell mass and the trophectoderm (ICM:TE ratio), and EGF-R protein expression in cloned embryos. In most instances these effects were enhanced in cloned embryos when EGF and TGF alpha were combined. Although fewer blastocysts developed from cloned than from fertilized one-cell stage embryos, growth factor treatment appeared to have the greatest effect on cloned embryos. These results demonstrate that mitogenic growth factors significantly enhance survival and promote the preimplantation development of cloned mouse embryos. PMID- 17907943 TI - Viral-mediated coexpression of Pdx1 and p48 regulates exocrine pancreatic differentiation in mouse ES cells. AB - Embryonic stem cells (ES) can spontaneously activate a pancreatic differentiation program in vitro, although with low efficiency. The aim was to improve such process by using viral mediated gene transduction. In this study, we have examined the suitability of using viral vectors to express key transcriptional factors involved in pancreatic development. ES cell lines that constitutively express Pdx1, a homeodomain protein involved in both exocrine and endocrine pancreatic development and differentiation, were established using a lentiviral vector. These cells were additionally infected with an adenovirus expressing p48, a bHLH factor that is also crucial for pancreatic development and acinar differentiation. Quantitative RT-PCR analysis demonstrated an increase in the expression of exocrine genes, including those coding for both digestive enzymes and transcription factors. Immunocytochemical staining also revealed an increase in the number of amylase-expressing cell clusters. However, other important genes involved in acinar cell maturation (i.e., Mist1) were not modulated under these conditions, suggesting that the cells display features of immature exocrine cells or because of an uncoupled gene expression of the exocrine differentiation program. Importantly, this effect was selective for the acinar lineage as the expression of a large set of endocrine markers remained unchanged. Therefore, combined expression of key genes involved in pancreatic development may be a promising approach to generate mature pancreatic exocrine cells. PMID- 17907944 TI - Lentiviral-RNA-interference system mediating homogenous and monitored level of gene silencing in human embryonic stem cells. AB - Genetic modifications of human embryonic stem cells (hESCs) that will efficiently promote stable homogenous gene silencing, and will also allow monitoring of the silencing level, may be invaluable for the study of function of genes in early human embryogenesis, differentiation, and maintenance of pluripotency of hESCs. RNA-mediated interference (RNAi) emerges as a highly efficient tool for specific knockdown of gene expression. Lentiviruses are efficient vectors for the delivery and stable expression of transgenes in hESCs. We sought to develop a lentiviral RNAi-based system that will efficiently induce homogenous gene silencing and will allow the monitoring of its relative level in hESCs. Dual-promoter lentiviral vectors coexpressing an RNAi cassette and a reporter gene were initially used for efficient and stable induction of heterogeneous levels of gene silencing in polyclonal hESCs. This step was further combined with the isolation of transduced clones with different homogenous levels of gene silencing. The level of silencing in each of the clones correlated and could be monitored by the level of expression of the vector's reporter transgene. Thus, our system allows easy identification of clones with relatively different homogenous levels of gene silencing. Our approach would be valuable for the study of function of genes, in particular those whose role in hESCs biology depends on their level of expression. PMID- 17907945 TI - Placental expression of major histocompatibility complex class I in bovine somatic clones. AB - Abnormally increased placental expression of major histocompatibility complex class I (MHC-I) molecules at the trophoblastic surface has been suggested previously to be the cause of early fetal loss in nuclear transfer (NT) bovine pregnancies. Here, we report the lack of expression of MHC-I at the trophoblastic surface at D30 and D60 and in placentomes from D60 to term in placentas obtained by NT from three different genotypes and by artificial insemination, whatever the outcome of the pregnancy. MHC-I expression was assessed by immunohistochemistry using four different antibodies, including a novel beta2-microglobulin antibody. The MHC-I type of the clones was established using reference strand-mediated conformation analysis (RSCA); however, since it proved problematic to type the recipient animals in the same way, outcome of pregnancy could not be related to MHC compatibility. In conclusion, the present study provides no evidence to support abnormal expression of MHC-I on the trophoblastic surface in clones as a major cause of fetal loss during pregnancy after NT. PMID- 17907946 TI - An epigenetic modifier results in improved in vitro blastocyst production after somatic cell nuclear transfer. AB - The present study was designed to examine the effect of trichostatin A (TSA), an inhibitor of histone deacetylase, on development of porcine cloned embryos. Our results showed that treatment of cloned embryos derived from sow oocytes with 50 nM TSA for up to 24 h after the onset of activation could significantly improve blastocyst yield compared to the control (46.4+/-4.6% vs 17.7+/-4.9% for treated and untreated embryos, respectively; p<0.05), whereas similar cleavage rate and total cell number per blastocyst were observed. In order to assess if the improvement is cell line specific, three cell lines were tested, and for all cell lines an enhancement in blastocyst development compared to their corresponding control was observed. Our data demonstrate that TSA treatment after somatic cell nuclear transfer in the pig can significantly improve the in vitro blastocyst production. PMID- 17907947 TI - Production of viable pigs from fetal somatic stem cells. AB - Fetal somatic stem cells (FSSCs) are a novel type of somatic stem cells that have recently been discovered in primary fibroblast cultures from pigs and other species. The goal of the present study was to produce viable piglets from FSSCs. NT complexes were prepared from both FSSCs and porcine fetal fibroblasts (pFF) to permit comparison of these two donor cell types. FSSCs from isolated attached colonies were compared with pFF in their ability to form blastocysts upon use in NT. Fusion and cleavage rates were similar between the two groups, while blastocyst rates were significantly higher when using pFF as donor cells. FSSCs of three different size categories derived from dissociation of spheroids yielded similar results. The use of FSSCs of 15-20 microm in size yielded similar cleavage and blastocyst rates as fetal fibroblasts. In the final experiment NT complexes produced from FSSCs were transferred to foster mothers. After transfer to prepubertal gilts, three of seven recipients established pregnancies and delivered seven piglets, of which three piglets were viable and showed normal development. Results for the first time demonstrate that FSSCs are able to produce cloned embryos, and that pregnancies can be established and viable piglets can be produced. PMID- 17907948 TI - Development of sheep androgenetic embryos is boosted following transfer of male pronuclei into androgenetic hemizygotes. AB - Androgenetic embryos are useful model for investigating the contribution of the paternal genome to embryonic development. Little work has been done with androgenetic embryo production in domestic animals. The aim of this study was the production of diploid androgenetic sheep embryos. In vitro matured sheep oocytes were enucleated and fertilized in vitro; parthenogenetic and normally fertilized embryos were also produced as a control. Fifteen hours after in vitro fertilization (IVF), presumptive zygotes were centrifuged and scored for the number of pronucleus. IVF, parthenogenetic, and androgenetic embryos (haploid, diploid, and triploid) were cultured in SOFaa medium with bovine serum albumin (BSA). The proportion of oocytes with polyspermic fertilization increased linearly with increasing sperm concentration. After IVF, there was no significant difference in early cleavage and morula formation rates between the groups, while there was a significant difference on blastocyst development between IVF, parthenogenetic, and androgenetic embryos, the last ones displaying poor developmental potential (IVF, parthenogenetic, and haploid, diploid, and triploid androgenetic embryos: 43%, 38%, 0%, 2%, and 2%, respectively). In order to boost androgenetic embryonic development, we produced diploid androgenetic embryos through pronuclear transfer. Single pronuclei were aspirated with a bevelled pipette from haploid or diploid embryos and transferred into the perivitelline space of other haploid embryos, and the zygotes were reconstructed by electrofusion. Fusion rates approached 100%. Pronuclear transfer significantly increased blastocyst development (IVF, parthenogenetic, androgenetic: Diploid into Haploid, and Haploid into Haploid: 42%, 42%, 19%, and 3%, respectively); intriguingly, the Haploid + Diploid group showed the highest development to blastocyst stage. The main findings of our study are: (1) sheep androgenetic embryos display poor developmental ability compared with IVF and parthenogenetic embryos; (2) diploid androgenetic embryos produced by pronuclear exchange developed in higher proportion to blastocyst stage, particularly in the Diploid Haploid group. In conclusion, pronuclear transfer is an effective method to produce sheep androgenetic blastocysts. PMID- 17907949 TI - Developmental arrest and cytoskeletal anomalies of rat embryos reconstructed by somatic cell nuclear transfer. AB - Many factors influence success rates in animal cloning by somatic cell nuclear transfer (SCNT), including cell cycle stage of donor cells and recipient oocytes, the procedure of micromanipulation, and the activation protocol. This study was conducted to determine the effects of cell cycle coordination for cloning rats from fetal fibroblasts (FFs). Moreover, enucleated zygotic and parthenogenetic ooplasts were used for serial cloning with pronuclear and two-cell stage blastomeres derived from SCNT. Metaphase donor cells had a significantly higher cleavage rate than G0/G1-phase FFs with MII oocytes and G2-phase FFs with TII oocytes. However, reconstructed embryos were unable to develop beyond the two cell stage, neither in vitro nor in vivo. Moreover, the developmental arrest at the two-cell stage was not overcome, even when using serial cloning with zygotic and parthenogenetic recipients. To assess the cytoskeleton after SCNT, reconstructed two-cell stage embryos were harvested at different times after cleavage for immunostaining (anti-alpha-tubulin) and mRNA abundance (beta-actin, alpha-tubulin, alpha-actinin). Reconstructed two-cell embryos showed abnormal microtubule distribution and down-regulated expression of several cytoskeletal transcripts. Therefore, it seems that the developmental arrest of rat SCNT embryos is associated with improper transcription of cytoskeleton genes, presumably resulting in abnormal microtubule distribution. PMID- 17907950 TI - Mouse fibroblasts are reprogrammed to Oct-4 and Rex-1 gene expression and alkaline phosphatase activity by embryonic stem cell extracts. AB - A recent remarkable study has shown that when mouse NIH-3T3 fibroblasts are exposed to an embryonic stem cell (ESC) extract, the majority of them expresses the Oct-4 gene, form ESC-like colonies, and embryoid-like bodies that differentiate into cells of the three germ layers. The use of cell extracts for inducing cell dedifferentiation could be a powerful system to obtain large quantities of pluripotent cells. It is thus of crucial importance that the robustness of this method of cell transdifferentiation is tested by other laboratories before it is advanced to a more ambitious use in cell therapy programs. We report here our experimental observations using the same reprogramming protocol on STO and NIH-3T3 mouse fibroblasts. Three are the main results: first, we confirmed an enduring reprogramming activity of the ESC extract, although on a much smaller number of cells that varies from approximately 0.003 to 0.04% of the total population of fibroblasts and with an effect limited to the induction of Oct-4 and Rex-1 gene expression and alkaline phosphatase activity. Second, the expression of OCT-4, SSEA-1, and Forssman antigen proteins was never detected. Third, our work has clearly demonstrated that ESCs may survive the procedure of extract preparation, may be source of contamination that is expanded in culture and give false positive results. PMID- 17907951 TI - Effects of growth factors on the differentiation of murine ESC into type II pneumocytes. AB - We have previously shown that embryonic stem cells (ESC) can be directed to differentiate into alveolar type II cells by provision of a serum-free medium designed for in vitro maintenance of mature alveolar epithelial cells (small airway growth medium: SAGM), although the target cell yield was low. SAGM comprises a basal serum-free medium (SABM) plus a series of defined supplements. In order to try increase the proportion of pneumocytes in differentiated cultures, we aimed in this study to determine the effects on murine ESC of each of the individual growth factors in SAGM. In accordance with our previous reports, expression of surfactant protein C (SPC) and its mRNA was used to monitor differentiation of type II pneumocytes. Surprisingly, we found that addition of each factor separately to SABM decreased the expression of SPC mRNA when compared with the effect of SABM alone. Thus, it seems that the observed enhancement by SAGM of pneumocyte differentiation from murine ESC can, in fact, be attributed to the provision of a serum-free environment. PMID- 17907952 TI - The effects of delayed activation and MG132 treatment on nuclear remodeling and preimplantation development of embryos cloned by electrofusion are correlated with the age of recipient cytoplasts. AB - The electrofusion method, used extensively in livestock cloning, cannot be used in mice, because it is believed that the mouse oocytes are more susceptible to detrimental effects of electrical stimulus than oocytes from other species. Reports on whether a delayed activation after electrofusion and a premature chromosome condensation (PCC) is essential for efficient cloning are inconclusive. To address these issues, effects of pulsing on activation and MPF activity of nonenucleated oocytes and effects of delayed activation and MG132 treatment on donor nuclear PCC and preimplantation development of embryos cloned by electrofusion or nuclear injection were compared between different cytoplast ages in mice and goats. The results indicated that the use of oocytes collected early after donor stimulation would make it possible to conduct somatic cell nuclear transfer in mice by electrofusion. Whether a delayed activation is essential was dependent upon the age, or rather, the level, of MPF activity of the cytoplasts at the time of electrofusion, as was the requirement for MG132 treatment. The competence for blastocyst formation of cloned embryos was highly correlated with the level of donor nuclear PCC in recipient cytoplasts. The nuclear injection technique was more adaptable to older cytoplast ages, and hence less dependent on drugs for inhibition of MPF inactivation, compared to electrofusion. PMID- 17907954 TI - Maintaining the trust of physicians and the public in the medical literature: report of a task force on scientific publishing of clinical trials. AB - In 2006, the American Society of Bone and Mineral Research and the Journal of Bone and Mineral Research convened a task force to consider whether and how to change our editorial policies to assure complete and unbiased reporting of clinical trials. We invited editors of journals that publish research on osteoporosis and disorders of bone and mineral metabolism and presidents of related societies to participate. The task force was charged to consider whether journals should (1) adopt the Principles for Protecting Integrity in the Conduct and Reporting of Clinical Trials published in 2006 by the American Association of Medical Colleges (AAMC) and should (2) require authors and sponsors of industry funded clinical trials to provide a jointly signed letter that states that the authors had full access to all the data and analyses on which the manuscript was based. The AAMC Principles recommend that multicenter trials should designate a Lead Investigator, Steering Committee, and Publication and Analysis (P&A) Committee, which should consist of a majority of academic investigators who are not sponsor employees. The P&A Committee should have the right to access any data generated during a study and to conduct its own statistical analyses. A majority of task force members voted to support the AAMC Principles, to require a letter jointly signed by academic investigators and industry sponsor stating that the authors had access to the data on which the submission was based, and to recommend adoption of these requirements to their respective societies and journals. Broad-based adoption of the AAMC Principles and requirement of a jointly signed attestation of data access by journals that publish clinical trials in diseases of bone and mineral metabolism should improve the position of academic clinical investigators in their interactions with industry and other funding sources. PMID- 17907956 TI - The road to LOAD: late-onset Alzheimer's disease and a possible way to block it. AB - The ageing brain becomes increasingly less able to destroy or eject toxic amyloid (A) beta42 peptide byproducts of normal neuronal activity that consequently accumulate to induce Alzheimer's disease (AD). Therefore, the various components of the Abeta-clearing machinery are prime targets for AD therapeutics. In this connection, there are reports that taking statins to lower circulating cholesterol to prevent cardiovascular disease can also prevent late-onset AD (LOAD) the most common form of the disease. However, it seems unlikely that statins would prevent LOAD by lowering the very long-lived brain cholesterol that is controlled independently from the very much shorter-lived circulating cholesterol. In fact, reducing the ability of the brain astrocytes to make cholesterol for their closely associated neuron clients' synaptogenesis could damage the brain rather than protect it. However, a plausible way statins might prevent LOAD is to target a main component of the clearance machinery, low density lipoprotein receptor-related protein 1 (LRP1), the brain's powerful Abeta efflux driver. This is indicated by a reported ability of micromolar concentrations of lovastatin and simvastatin to strongly stimulate brain vascular endothelial cells to make this Abeta ejector. Therefore, if this holds up, taking a statin over the years would prevent the normal decline of LRP1 in the ageing brain and a LOAD-driving accumulation of Abeta. PMID- 17907957 TI - Interleukin-18: a pro-inflammatory cytokine that plays an important role in acute pancreatitis. AB - A large body of clinical and experimental evidence suggests that cytokines play a key role in the pathogenesis of local and systemic complications of acute pancreatitis. IL-18 is a pro-inflammatory cytokine that plays a key role in many human diseases, including acute pancreatitis. This review focuses on the present understanding in IL-18 and its potential role in acute pancreatitis. IL-18 levels reflect the severity of acute pancreatitis and display a significant negative correlation with the concentrations of antioxidative damage factors, serum selenium and glutathione peroxidases (GPx). The relationship between IL-18 and other pro-inflammatory cytokines shows that IL-18 is one of the key mediators of inflammation in the pathogenesis of acute pancreatitis. Elevation of serum IL-18 levels may mediate acute pancreatitis associated liver injury. The use of IL-18 antagonists as direct routes to block IL-18 activity and P2X7 receptor antagonists and interleukin-1beta-converting enzyme (ICE) inhibitors as indirect routes to block IL-18 activity suggest that specific therapeutic inhibition of IL 18 is a promising therapeutic approach for acute pancreatitis. PMID- 17907958 TI - Targeting systemic inflammation: novel therapies for the treatment of chronic obstructive pulmonary disease. AB - The increasing evidence that inflammation in the lungs leads to the structural changes observed in chronic obstructive pulmonary disease, whereas extrapulmonary symptoms and comorbidities may be systemic manifestations of these inflammatory processes, highlights an urgent need to discover novel, effective anti inflammatory treatments for this disease. Some studies are suggesting that, by decreasing dynamic hyperinflation, bronchodilators might reduce systemic inflammation; inhaled corticosteroids and their combination with long-acting beta2-agonists might contribute to this goal. Even so, the opinion that suppression of the inflammatory response might improve systemic complications is stimulating a search for novel anti-inflammatory therapies. Many drugs include those that inhibit the recruitment and activation of inflammatory cells and/or antagonise their products. However, many of these therapeutic strategies are not specific for neutrophilic inflammation because they affect other cell types, thus, it is difficult to interpret whether any clinical benefit observed is a result of a reduction in airway neutrophils. In any case, there is some evidence that drugs used to treat a co-morbid condition, such as statins, angiotensin converting enzyme (ACE) inhibitors and angiontensin II type 1 (AT1) receptor blockers as well as glycosaminoglycans and peroxisome proliferator-activated receptor (PPAR) agonists, might benefit chronic obstructive pulmonary disease patients because they deal with the extrapulmonary, systemic component of chronic obstructive pulmonary disease. PMID- 17907959 TI - Tackling EGFR signaling with TACE antagonists: a rational target for metalloprotease inhibitors in cancer. AB - TNF-alpha converting enzyme (TACE/ADAM17) is a transmembrane metalloprotease that plays a key role in the cleavage and mobilization of receptor ligands that are initially synthesized as membrane-tethered precursors. For many years, attention has focused on the role of TACE-dependent TNF-alpha cleavage in arthritis and, more recently, it has become apparent that TACE also plays an important role in regulating epidermal growth factor receptor activity in several tumor types. This review presents the background to these findings and a rationale for the continued development of TACE inhibitors for the treatment of epidermal growth factor receptor-dependent epithelial tumors. PMID- 17907961 TI - Focal adhesion kinase: a promising target for anticancer therapy. AB - Focal adhesion kinase (FAK) is a protein tyrosine kinase acting as an early modulator of the integrin signalling cascade, thus regulating various basic cellular functions. In transformed cells, upregulation of FAK protein expression and uncontroled signalling were held responsible for the promotion of malignant phenotypic characteristics, as well as resistance to chemotherapy and radiotherapy. Direct FAK targeting resulted in the inhibition of the malignant phenotype of cancer cells, whereas increased apoptotic rates of cancer cells, either used alone or in combination with conventional chemotherapeutic agents, radiotherapy or hormonal therapy. Furthermore, drugs used in cancer chemotherapy, besides their basic mode of action, were also shown to act through altering FAK signalling. Finally, positive results were noted by the transfection of cancer cells with fak mutants or genes that suppress FAK expression or activity, such as phosphatase and tensin homolog deleted on chromosome Ten (PTEN), ribonucleotide reductase M1 polypeptide (RRM1) and melanoma differentiation-associated gene-7 (mda-7). The purpose of this article is a comprehensive review of the existing data on the possible use of FAK targeting in anticancer therapy. PMID- 17907962 TI - Targeting PGC-1 alpha to control energy homeostasis. AB - The prevalence of Type 2 diabetes is increasing at an alarming rate in most parts of the world. Effective therapeutic drugs are urgently needed, not only to control the disease but also to prevent or delay its progression. Therapies that target the underlying pathogenesis could, in theory, hold such potential. Recent evidence strongly suggests that impaired mitochondrial function is part of the underlying pathogenesis of insulin resistance and Type 2 diabetes. Peroxisome proliferator-activated receptor gamma co-activator-1 alpha (PGC-1alpha) is a transcription co-activator that plays a key role in regulating mitochondrial biogenesis and energy metabolism in multiple tissues. Thus, improvement and restoration of mitochondrial function and oxidative capacity through activation of PGC-1alpha could provide new treatments for metabolic diseases. A diverse array of proteins has been shown to regulate PGC-1alpha transcription and/or activity, some of which represent promising targets for pharmaceutical intervention. PMID- 17907963 TI - c-Jun N-terminal kinases as potential therapeutic targets. AB - One principal aim of research in the signal transduction field is to identify targets for therapeutic intervention, in an attempt to modify disease and curtail human suffering. Diseases such as chronic inflammation, atherosclerosis, diabetes and cancer exact a huge toll on health, in physical, social and financial terms. Defective signaling mechanisms are central to their pathogenesis. One candidate signaling molecule that is presently undergoing intense investigation is the c Jun N-terminal kinase. With roles described in almost all classes of disease, the main questions are what type of inhibitor to use and when exactly to use it during the disease course? PMID- 17907964 TI - STAT nuclear translocation: potential for pharmacological intervention. AB - The signal transducer and activator of transcription (STAT) proteins are extracellular ligand-responsive transcription factors that mediate broadly diverse biological processes, including cell proliferation, transformation, apoptosis, differentiation, fetal development, inflammation and immune response. Stimulation with multiple cytokines or growth factors all result in the tyrosine phosphorylation of STAT proteins and the subsequent gene regulation via their direct binding to the promoters of responsive genes. Cytokine-regulated gene activation is dependent on the continuous nucleocytoplasmic cycling of STAT signal transducers. The STATs use intricately intertwined karyopherin-dependent and -independent translocation mechanisms to coordinate the activation step at the cell membrane and gene expression in the nucleus. In addition, STATs appear to have cytokine-independent gene regulatory functions that may also depend on their regulated nucleocytoplasmic transfer. Numerous studies have implicated aberrant STAT signalling in cancer, immune defects and inflammatory diseases. Given the central role of intracellular trafficking for the proper signal processing by STAT proteins, pharmacological targeting of STAT nucleocytoplasmic translocation appears to be an attractive strategy to interfere with dysregulated cytokine signalling. This review will discuss possible scenarios that would result from the use of novel modulators of STAT shuttling, which may both increase or decrease STAT activation and, hence, transcriptional activity. PMID- 17907960 TI - TRAIL in cancer therapy: present and future challenges. AB - Since its identification in 1995, TNF-related apoptosis-inducing ligand (TRAIL) has sparked growing interest in oncology due to its reported ability to selectively trigger cancer cell death. In contrast to other members of the TNF superfamily, TRAIL administration in vivo is safe. The relative absence of toxic side effects of this naturally occurring cytokine, in addition to its antitumoural properties, has led to its preclinical evaluation. However, despite intensive investigations, little is known in regards to the mechanisms underlying TRAIL selectivity or efficiency. An appropriate understanding of its physiological relevance, and of the mechanisms controlling cancer cells escape from TRAIL-induced cell death, will be required to optimally use the cytokine in clinics. The present review focuses on recent advances in the understanding of TRAIL signal transduction and discusses the existing and future challenges of TRAIL-based cancer therapy development. PMID- 17907966 TI - Expression of pIX gene induced by transgene promoter: possible cause of host immune response in first-generation adenoviral vectors. AB - First-generation (FG) adenoviral vectors (AdVs) have been widely used not only for gene therapy but also for basic studies. Because vectors of this type lack the E1A gene that is essential for the expression of other viral genes, their expression levels in target cells have been considered low. However, we found that the viral pIX gene, located immediately downstream of the inserted expression unit of the transgene, was significantly coexpressed with the transgene in cells infected with FG AdV. Whereas CAG and SRalpha promoters activated the pIX promoter considerably through their enhancer effects, the EF1alpha promoter hardly did. Moreover, when the expression unit was inserted in the rightward orientation, not only the pIX protein but also a fusion protein consisting of the N-terminal part of transgene product and pIX were sometimes coexpressed with the transgene product through an aberrant splicing mechanism. In in vivo experiments, a LacZ-expressing AdV bearing the CAG promoter caused an elevation of alanine aminotransferase, but an AdV bearing the EF1alpha promoter produced no detectable levels. Whereas the FG AdV expressing human growth hormone under the control of the CAG promoter maintained a high hormone level for less than 1 month, the FG AdV under the control of the EF1alpha promoter maintained a high level for at least 6 months. These results suggest that pIX coexpression may be one of the main causes of AdV-induced immune responses, and that the EF1alpha promoter is probably valuable for the long-term expression of FG AdV. Thus, the in vivo utility of FG AdV should be reevaluated. PMID- 17907967 TI - Retroviral integration site analysis and the fate of transduced clones in an MDR1 gene therapy protocol targeting metastatic breast cancer. AB - A clinical study of an MDR1 gene therapy protocol targeting metastatic breast cancer has been conducted in which the patients received high-dose chemotherapy, a transplant of MDR1-transduced autologous CD34(+) cells, and docetaxel. We herein report the molecular results of a 6-year follow-up of an individual in this study (patient 1). HaMDR-transduced cells, which had been initially detected in the peripheral blood of this individual, were found to have gradually decreased. After 10 cycles of docetaxel (days 71-316), MDR1 transgene levels were found to have increased, and then decreased to undetectable levels by day 1461. Thirty-eight MDR1-transduced clones were identified in patient 1, of which 11 showed a retroviral integration in close proximity to genes listed in the Retrovirus Tagged Cancer Gene Database (RTCGD). Four short-life clones in this group were found to harbor retroviral integrations close to the ZFHX1B, NOTCH1, BMI1, or HHEX gene; these genes have been frequently reported in the RTCGD. In addition, a long-lived RTCGD-hit clone, L-34, had a retroviral integration at a position 179 kb upstream of the EVI1 gene. L-34 was detectable on days 327-1154, but became undetectable 3 years after the docetaxel treatments had ceased. An additional three docetaxel-induced long-life clones showed comparable polymerase chain reaction profiles, which were also similar to that of the total MDR1 transduced cells. Our results thus show that docetaxel may have been effective in promoting the expansion of several MDR1-transduced clones in patient 1, but that they persist in the peripheral blood for only a few years. PMID- 17907968 TI - Laparoscopic totally extraperitoneal hernia repair versus open Lichtenstein hernia repair: results and complications. AB - Whereas open Lichtenstein inguinal herniorrhaphy is generally accepted as a safe, well-understood method with a high success rate, the laparoscopic repair of a inguinal hernia is a fairly recent technique. Although the laparoscopic approach to a hernia repair procedure is associated with less pain and faster recovery than open repair, many surgeons are not familiar with this technique owing to technical demands and a long learning curve. This study compares the results and complications between open tension-free mesh (Lichtenstein) repair and laparoscopic total extraperitoneal (TEP) repair. The study cohort was comprised of 345 consecutive patients who underwent an inguinal herniorraphy procedure. An open hernia repair was performed on one group of patients (n = 233), whereas TEP repair was performed on the other (n = 112), and then the comparison of intra- and postoperative complications and results obtained from both techniques was done. The mean hospital stay was similar in both groups. The average operative time in the TEP group was 58.6 +/- 18.1 minutes, and the average operative time in the open group was 58.2 +/- 17.8 minutes. There was no difference in postoperative complication rates between the two groups, except for urinary retention, which patients who underwent TEP repair were more likely to get. The following major complications were recorded: 2 cases of urinary bladder perforation-1 during TEP repair and the other during Lichtenstein repair, but both with good postoperative outcome-and 1 case of pneumothorax, which occurred during the TEP procedure. Despite the fact that TEP is a demanding procedure, it may be performed efficiently with an acceptable operating time and a low complication rate. PMID- 17907969 TI - Laparoscopic approach to recurrent incisional hernia repair: a 3-year experience. AB - BACKGROUND: Incisional hernias are one of the most frequent complications of open abdominal surgery. The incidence of relapses after a conventional repair procedure is higher in recurrent than in primary cases (30%-50% vs. 11%-20%). The laparoscopic approach can prevent the complications associated with the conventional approach when dealing with recurrent incisional hernias. The aim of this study was to evaluate the efficacy of laparoscopic treatment in such cases. MATERIALS AND METHODS: We prospectively analyzed data from 41 consecutive patients with recurrent incisional hernias, who submitted to a laparoscopic repair procedure with an expanded polytetrafluoroethylene Dual Mesh (Gore-Tex Dual Mesh Plus Biomaterial; W.L. Gore 8 Associates) from December 2001 to December 2004. All of the patients underwent clinical follow-up at 1, 6, and 12 months and then yearly. An ultrasound scan of the abdominal wall was performed at 6 and 12 months after the procedure. The parameters considered for the analysis were: mesh size, operating time, hospital stay, postoperative complications, and recurrences. RESULTS: The defects were usually localized along midline laparotomies. The mean mesh size was 400 cm2, the mean operating time was 68 minutes, and the mean length of hospital stay was 2.7 days. Complications were encountered in 17% of patients. The mean follow-up was 38 months (range, 18-54). Recurrence was reported in 1 case only (2.4%), which occurred within the first 6 months after the operation. CONCLUSIONS: The laparoscopic repair of recurrent incisional hernia seems to be an effective alternative to the conventional approach, as it can give lower recurrence and complication rates. PMID- 17907970 TI - Laparoscopic-assisted placement of peritoneal dialysis catheters: implantation technique and results. AB - BACKGROUND: Peritoneal dialysis is a generally accepted method for the treatment of patients with end-stage renal failure. The laparoscopic placement of peritoneal dialysis catheters is a well-established technique and offers some advantages, such as a safer placement of the catheter, less post-operative complications, and a longer functional survival, compared to the conventional open technique. The aim of this study was to describe our implantation technique and to determine the results of our approach. PATIENTS AND METHODS: Between January 2000 and February 2006, 47 patients with end-stage chronic renal failure underwent a laparoscopic peritoneal dialysis catheter insertion procedure. Perioperative and follow-up data were collected prospectively. RESULTS: The mean operating time was 35 minutes (range, 16-100). There was no perioperative morbidity. Nine (19.1%) patients experienced 10 mechanical complications: fluid leakage in 6 (12.8%) patients, acute hydrothorax in 1 (2.1%), catheter tip migration in 2 (4.3%), and catheter obstruction in 1 (2.1%) patient. Episodes of peritonitis were observed in 5 (10.6%) patients. One (2.1%) patient developed a catheter infection. In 3 (6.4%) patients, a port site hernia occurred that required surgical repair, 5 (10.6%) patients underwent laparoscopic revisions owing to mechanical complications, 9 (19.1%) patients underwent renal transplantation, and 6 (12.8%) patients died during the later follow-up. After a mean follow-up time of 17 months (range, 2-76), 30 (63.8%) catheters are still in use for dialysis. CONCLUSIONS: The functional outcome of the dialysis catheters was satisfactory in the majority of patients in this study. The described technique for catheter implantation is simple and safe, and in our opinion, the laparoscopic technique should be considered as the method of choice in patients with end-stage chronic renal failure. PMID- 17907971 TI - Trocar site hernia after laparoscopic cholecystectomy. AB - AIM: The aim of this study was to elucidate the influence of pre and perioperative factors on the development of trocar site hernia after a laparoscopic cholecystectomy procedure. PATIENTS AND METHODS: A total of 776 patients who underwent a laparoscopic cholecystectomy procedure in our Department of General Surgery between 1999 and 2004 were assigned as the study group. The control group included patients without trocar site hernias after a cholecystectomy. The effect of five variables, including gender, age, body mass index (BMI), operation duration, and the type of cholecystitis on the development of a trocar site hernia after a laparoscopic cholecystectomy was assessed by univariable and multivariable models. RESULTS: In the univariate analysis, female gender (P = 0.021), older age (P < 0.001), higher BMI at the time of surgery (P < 0.001), and an increased duration of surgery (P < 0.001) have been found to increase the likelihood of a trocar site hernia formation. However, in the multivariable model, the gender was not a significant variable to influence the development of this complication. CONCLUSIONS: The development of a postoperative trocar site hernia may be prevented by the closure of 10-mm trocar sites in patients who are older than 60 years, obese, and who have a longer duration of operation. PMID- 17907972 TI - Low incidence of adhesion-related bowel obstruction after laparoscopic colorectal surgery. AB - BACKGROUND: Postoperative adhesions are a major cause of morbidity, accounting for approximately 5% of the readmissions of surgical patients. Bowel obstruction is attributed to adhesions in more than half of the cases, many of which are following colon and rectal surgery. Laparoscopic surgery has the potential advantage of reduced adhesion formation owing to attenuated surgical trauma, less tissue handling, and smaller scars. However, the translation of these advantages to a reduced rate of bowel obstruction has not been sufficiently demonstrated. The aim of this study was to assess the rate of adhesion-related bowel obstruction after laparoscopic colon and rectal surgery. METHODS: Data regarding all cases of laparoscopic colon and rectal surgery were prospectively collected. Information relative to demographics, surgical procedures, and follow-up was analyzed, and patients who were readmitted for bowel obstruction were identified. RESULTS: Over a period of 8 years, 306 patients, at a mean age of 63 years, had a laparoscopic colon and rectal operation in our department-122 for benign conditions and 184 for malignant disease. The mean length of follow-up was 38 months. Six cases (2%) of bowel obstruction, which were unrelated to hernia or advanced cancer, were identified. Two patients had a history of open surgery, in addition to the laparoscopic procedure, so adhesions could be attributed solely to the laparoscopic procedure in 4 patients, which consisted of 1.3% of the total study group. Obstruction occurred within 2 weeks of surgery in 2 patients, and one early reoperation was required. CONCLUSIONS: The incidence of adhesion ileus after laparoscopic colon and rectal surgery appears to be very low. This long term benefit of laparoscopic surgery should be considered when comparing this technique to its open counterpart. PMID- 17907973 TI - Single-center experience of laparoscopic cholecystectomy. AB - BACKGROUND: The laparoscopic cholecystectomy procedure is considered as the gold standard for the management of benign symptomatic gallbladder diseases. In this paper, we present our experience as a tertiary reference center in the management of this disease. METHODS: A total of 9864 laparoscopic cholecystectomies have been performed in our institution since 1991. All patients undergo a routine hematologic work-up, high resolution ultrasonography, and, thereafter, a four port cholecystectomy by the North American approach. We specifically considered 10 areas of controversy. RESULTS: The male:female ratio was 45:55, with the average age being 40.4 years (range, 1 year 4 months to 92 years). Asymptomatic patients were 986 in number and 25% had acute cholecystitis, whereas 2.28% had associated choledocholithiasis. The "fundus first" approach was adopted in 88 patients. Three patients had occult gallbladder cancer, of which 2 were in situ and 1 was of the T1 stage. Nearly one third of the patients had gallstone spillage, primarily owing to the performance of a subtotal cholecystectomy procedure in the setting of acute cholecystitis. However, only 1 patient to date has had a problem directly attributable to gallstone spillage. Nearly 13% of patients had an additional procedure along with a cholecystectomy. The average operating time was 21.5 minutes, whereas the average length of postoperative stay was 1.6 days. CONCLUSIONS: The laparoscopic approach is suitable for the management of all forms of benign gallbladder diseases. In addition, it is possible to use the laparoscopic approach for indications hitherto considered relatively contraindicated. In a dedicated center, it is possible to perform a large number of cases with low morbidity and minimal conversion rates. PMID- 17907974 TI - Laparoscopic repair of peptic ulcer perforation without omental patch versus conventional open repair. AB - BACKGROUND: Laparoscopic surgery, a minimally invasive technique, has recently begun to be used on perforated peptic ulcers effectively and frequently. Nevertheless, most studies have shown that the disadvantages of the laparoscopic treatment of peptic ulcers are a long operation time, a high reoperation rate, and a need for an experienced surgeon. Thus, the objective of the current study was to compare the safety and efficacy of optimized laparoscopic surgery without an omental patch for a perforated peptic ulcer within a shorter operational time with conventional open surgery in a 4-year period. PATIENTS AND METHODS: From May 2002 to June 2006, 35 consecutive patients with a clinical diagnosis of a perforated peptic ulcer were prepared prospectively to undergo either an open or optimized laparoscopic surgery. RESULTS: Seventeen patients with a perforated peptic ulcer underwent simple laparoscopic repair without an omental patch. Three patients (17.6%) who were begun by the laparoscopic approach had to be converted to open surgery. Eighteen patients underwent conventional open surgery. The mean operative time for laparoscopic repair was 42.10 minutes (range, 35-60), which was significantly shorter than the 55.83 minutes for open repair (range, 35-72; P = 0.001). Postoperative parenteral analgesic requirements were lower after laparoscopic repair (75.0 mg) than that after an open repair procedure (101.39 mg; P = 0.02). There was no statistically significant difference between the procedures in terms of hospital stay (5 vs. 5.33 days; P = 0.37) and the timing of access to normal daily activity (6.8 vs. 7.1 days) (P = 0.54). CONCLUSIONS: Laparoscopic surgery, when optimized by a simple repair without an omental patch and 10 mm of a large-channel aspirator-irrigator, may be safely and effectively applied to the patients with small duodenal perforated peptic ulcers (<10 mm) and because of its having low risk factors. The procedure may be an alternative treatment to other procedures when in experienced hands. PMID- 17907975 TI - Laparoscopic cholecystectomy without intraoperative cholangiography. AB - BACKGROUND: The aim of this retrospective study was to evaluate the outcome of laparoscopic cholecystectomies (LCs) performed in our Academic Surgical Unit, and the impact of our policy not to perform intraoperative cholangiograms (IOCs) on the incidence of bile duct injuries (BDIs). MATERIALS AND METHODS: Data was collected for the time period from 1992 (when the laparoscopic procedure was first introduced in our Unit) until 2005. During this time, 1851 patients underwent an LC. Patients with a history of jaundice, ultasonographic bile duct dilatation, bile duct stones, or deranged liver function tests were referred initially for an endoscopic retrograde cholangiopancreatography procedure. An IOC was not performed on any patient. RESULTS: The conversion rate was 23.9% among the patients with acute cholecystitis and 1.6% among the patients with a noninflamed gallbladder. This difference was statistically significant. The morbidity reached 1.1%, as minor or major complications were present in 22 of 1851 patients. Complications consisted of BDI in 7 patients (0.37%). Six patients presented with minor BDI. Two of the BDIs occurred among the group of patients with acute cholecystitis, whereas the remaining 5 occurred in the group of patients with a noninflamed gallbladder. This distribution was not statistically significant. CONCLUSIONS: The low BDI rate in our series allowed us to recommend an LC procedure without an IOC. Performing a cholangiogram either routinely or selectively is not wrong. However, adherence to a meticulous hemostatic technique, thorough knowledge of the anatomy, and a low threshold for conversion may also enable satisfactory results to be achieved. PMID- 17907976 TI - Laparoscopic management of acutely presenting gastrointestinal stromal tumors: a study of 9 cases and review of literature. AB - INTRODUCTION: Gastrointestinal stromal tumors (GISTs) are rare. Nevertheless, some may present with a life-threatening hemorrhage or intestinal obstruction. The aim of this study was to review the various modes of the presentation of GIST, especially hemorrhage and obstruction, and to assess the role of laparoscopic surgery in the management of acute cases and the correlation of such cases with malignant potential. METHODS: Data were collected from the hospital in the patient records as well as a prospective database. Their presentation, management, histologic features, and follow-up periods were analyzed. RESULTS: Nine cases of GIST were collected. Five (56%) presented with hemorrhage, 3 (33%) with intestinal obstruction, and 1 (11%) with a tender epigastric mass. Six patients had a gastroscopy procedure, and 6 had a computed tomography scan of the abdomen and pelvis. Five (67%) patients underwent a partial gastrectomy (3 laparoscopic and 2 open procedures), 1 had a laparoscopic localized resection of a gastric GIST, and 3 (33%) had a resection and anastomosis of the bowel. All patients presented acutely, and 78% had emergency surgery performed. The laparoscopic approach was attempted in 6 patients (67%), and was successful in 4 (67%). All patients who had a laparoscopic treatment performed had less pain, a quicker recovery, and a shorter hospital stay, as compared to those patients who had open surgery. Five cases were malignant tumors. Immunohistochemistry revealed a positive c-kit and CD34 for all tumors. The median follow-up was 24 months with one recurrence. CONCLUSIONS: Our experience showed that GISTs can present acutely and may need immediate surgical intervention. A laparoscopic treatment is safe and practical in experienced hands. Tumor size and hemorrhage at presentation can predict a patient's malignant potential. PMID- 17907977 TI - Efficacy and safety of early laparoscopic common bile duct exploration as primary procedure in acute cholangitis caused by common bile duct stones. AB - INTRODUCTION: The elective laparoscopic management of common bile duct (CBD) stones is widely accepted; however, the urgent laparoscopic exploration of common bile duct (LCBDE) within the first 72 hours of acute cholangitis is not assessed extensively. Our aim was to study the safety and efficacy of urgent LCBDE in patients with acute cholangitis. MATERIALS AND METHODS: In a single-center prospective study, 73 patients of a university hospital with acute gallstone cholangitis were operated on with laparoscopy or open surgery, based on a predetermined schedule concerning the presence of the skilled laparoscopic surgeon at the hospital. Patients with sever acute cholangitis (e.g., organ failure, shock, or peritonitis), pancreatitis, and suspected tumoral obstructions were excluded. The major outcomes, including mortality, complications of surgery, and the length of hospital and intensive care unit (ICU) stay, are reported in this paper. RESULTS: In all 36 open surgery patients, a choledocotomy and T-tube placement procedure were performed. In laparoscopic patients, CBD clearance was approached by a transcystic and choledocotomy approach in 15 and 22 subjects, respectively. Eight (6 in the open and 2 in the laparoscopic group) choledocoduodenostomies were performed. Cholangitis was controlled sufficiently in all patients. Of 37 laparoscopies, 3 operations were converted into open surgeries. Operation time was longer in the laparoscopic group, compared to the open group (201 +/- 15 vs. 146 +/- 6.1 minutes; P < 0.01). The average ICU and hospital stay after an operation were significantly less than open surgery group. Total cost of treatment in laparoscopic group was less than 75% of that of the open surgery group. General complications were more common in the open surgery group. There was no mortality. One retained stone was discovered in the laparoscopic group. CONCLUSIONS: Early one-stage LCBDE is an effective procedure as an initial and definite management of acute gallstone cholangitis, which prevents a second hospitalization and relapse problems. PMID- 17907978 TI - Endolaparoscopic approach in the management of gastroesophageal reflux disease: an experimental study in pigs. AB - INTRODUCTION: Chronic reflux of gastric content to the esophagus causes retrosternal burning, pain, and regurgitation, and results in histopathologic changes that may culminate in adenocarcinoma. Insufficiency of the lower esophageal sphincter and hiatal hernia are the two principal causative factors. In this paper, we present the early results of a new antireflux operation in pigs. MATERIALS AND METHODS: The status of physiologic reflux was determined in 5 pigs (approximately 40-45 kg) with 24-hour pH monitorization. Under endoscopic guidance, a trocar was inserted into the stomach (similar to the percutaneous endoscopic gastrostomy technique). The endoscope was put into the retroflexed position; a mucomuscular pursestring suture was placed around the endoscope at the cardia and tied. Ten days later, pH monitorization was repeated to evaluate the effectiveness of the method. Late postoperative results have been reevaluated by 24-hour pH monitorization in the sixth month. RESULTS: The procedure achieved statistically significant improvements in total reflux time, number of reflux periods, number of long reflux periods, the longest reflux period, and the DeMeester score. The preoperative and postoperative DeMeester scores of the individual animals were 179.24-0.94, 11.48-0.98, 68.4-3.74, 132.2-46.49, and 38.72-5.86, respectively. Even though there appeared to be a slight increase in the pH monitorization levels after 6 months, these results did not reach significant degrees, and compared to the physiological reflux, the results were remarkable. CONCLUSIONS: The results obtained showed considerable decreases in physiologic reflux in all animals. We believe that this endoluminal, laparoendoscopic operation can be used in the treatment of gastroesophageal reflux disease as a minimally invasive method. PMID- 17907979 TI - A naval surgeon's approach to the draining umbilicus. AB - Spontaneous umbilical drainage is an uncommon problem in the adult, for which various congenital or acquired conditions may be responsible. In this paper, we present 3 cases demonstrating an approach that avoids the need for expensive diagnostic imaging. A careful office exam and basic laparoscopic skills are all that is needed. A brief discussion and review of the literature follows. PMID- 17907980 TI - Benign peritoneal multicystic mesothelioma diagnosed and treated by laparoscopic surgery. AB - Benign cystic mesothelioma is a rare pathology predominantly encountered in females. The increased use of laparoscopy for abdominal pain, particularly in female patients, implies that surgeons are aware of the macro- and laparoscopic presentation of this tumor for adequate diagnosis and therapy. In this paper, we present the case of a young woman with benign multicystic mesothelioma in which only laparoscopy led to the appropriate diagnosis. Subsequently, the tumor was removed by laparoscopic surgery. PMID- 17907982 TI - Laparoscopic cholecystectomy after open cholecystostomy for gallbladder empyema: a case report. AB - The case of a patient with gallbladder empyema initially drained through a minilaparotomy procedure under local anesthesia with a tube cholecystostomy is reported in this paper. Eight weeks later, the patient underwent an elective interval laparoscopic cholecystectomy. At laparoscopy, the gallbladder and the cholecystostomy tube were dissected free from the abdominal wall and the greater omentum, which was attached to the gallbladder. The tube was removed from the gallbladder fundus, and the operation was completed laparoscopically without any major problems. PMID- 17907981 TI - Bilateral pneumothorax after extraperitoneal laparoscopic radical prostatectomy. AB - A known complication of laparoscopic surgery is pneumothorax. Most reported pneumothoraces occur during laparoscopic transperitoneal abdominal surgery. Clinically significant pneumothorax from pelvic extraperitoneal surgery has not been reported. In this paper, we describe symptomatic bilateral pneumothorax following a totally extraperitoneal radical prostatectomy. The management of the complication is also discussed. PMID- 17907983 TI - A new technical approach for extraperitoneal laparoscopic bladder diverticulectomy. AB - In this paper, the authors report on a new, modified laparoscopic technique to remove a large bladder diverticulum. A 26-year-old male with a urinary problem underwent an ultrasound, as well as intravenous urography and cystoscopy examinations, which showed a large bladder diverticulum. The diverticulum was operated upon laparoscopically. The extraperitoneal laparoscopic intervention was facilitated by balloon placed into the diverticulum. The new technique for the laparoscopic diverticulumectomy procedure was successful and the operating time was 140 minutes. There were no perioperative complications. In conclusion, the laparoscopic removal of the bladder diverticulum is a safe and minimally invasive intervention. The introduction of a balloon into the diverticulum makes the operation easier. PMID- 17907984 TI - A case of cystic duct drainage into the left intrahepatic duct and the importance of laparoscopic fundus-first cholecystectomy for prevention of bile duct injury. AB - The laparoscopic cholecystectomy is the gold standard for the treatment of cholecystolithiasis, although it has been reported that the incidence of bile duct injury is higher for this method than for conventional open surgery. These injuries are mainly attributable to a misunderstanding of the biliary tract anatomy owing to severe cholecystitis, large impacted stones, and anatomic variations. In this paper, we report on the successful treatment of a 59-year-old male patient with cholecystolithiasis accompanied with extremely unusual biliary junction, in which the cystic duct drained into the left hepatic duct while using the laparoscopic fundus-first-cholecystectomy approach (i.e., the doom down technique) without any serious intraoperative complications. If the doom-down technique was not indicated in this particular case, the authors believe that a careless division of the cystic duct may have resulted in operative morbidity. PMID- 17907985 TI - Small bowel obstruction due to broad ligament hernia successfully treated by laparoscopy. AB - We present the second case of a broad ligament hernia to be treated by laparoscopy. A 43-year-old gravida 0, para 0 woman presented to our hospital with a chief complaint of right upper quadrant abdominal colicky pain and vomiting. Her clinical history was significant for an "unknown" bowel surgery through a Pfannenstiel incision. Her abdomen was soft, nondistended, and slightly tender to palpation in the right upper quadrant. The laboratory tests showed an elevated white blood cell count of 15.2 [1] 109/L with a left shift, and a normal serum C reactive protein. Plain abdominal X-rays showed a lightly prominent small bowel loop in the pelvic area. An abdominal ultrasound revealed only a small amount of fluid in the pouch of Douglas. After 24 hours, the pain had migrated to the hypogastrium. There was an increase in the white blood cell count, in the C reactive protein level, and in her temperature (37.7 degrees C). At this stage, we elected to perform an urgent laparoscopic exploration. We discovered 60-80 cm of ileum strangulated through a 2 x 3 cm defect in the left broad ligament of the uterus. The strangulated bowel was successfully reduced and the defect was closed with a 2-0 silk running suture. No bowel resection was required. The patient was discharged from the hospital on day 4, with no postoperative complications. PMID- 17907986 TI - Mesh erosion into caecum following laparoscopic repair of inguinal hernia (TAPP): a case report and literature review. AB - Repair of inguinal hernia is the most commonly performed surgical procedure. Both open and laparoscopic methods are accepted modalities of surgical treatment. Transabdominal preperitoneal (TAPP) and total extraperitoneal (TEP) are the two types of laparoscopic repair of the inguineal hernia. The main advantages of laparoscopic repair, as compared to open repair, are a shorter hospital stay and a quicker recovery to normal activities. However, laparoscopic repairs are associated with a higher incidence of visceral and vascular injuries. One particular complication is the migration and erosion of mesh into the adjacent viscera. Although the total numbers of cases are small, compared to the total numbers of inguinal hernia repairs, they are important, as they often presented with a diagnostic dilemma. Most of the mesh migrations reported in the literature involves the urinary bladder. In this paper, we present a case of erosion of mesh into the caecum. The patient (a 66-year-old male) underwent TAPP repair of a right inguinal hernia in 1996 with polypropelene mesh. He also underwent an open appendicectomy in 1980. During the laparoscopic repair, he was found to have multiple intra-abdominal adhesions. He presented with intermittent diarrhea, for which he was investigated, and a benign caecal lesion was found. He was initially managed conservatively. However, his symptoms persisted and he underwent a right hemicolectomy in February 2006 in our hospital. The offending lesion was found to be the prolene mesh having eroded into the caecum. PMID- 17907987 TI - Telesurgery system with original-quality moving images over high-speed Internet: expansion within the Asia-Pacific region. AB - BACKGROUND: Amid rapid changes in surgical techniques and patient care, education and training for the new generation of health care providers is of utmost importance. An international telesurgical system, which we established between Japan and Korea through super-fast broadband Internet without any loss of quality, was shown to be a powerful tool for this purpose. We attempted to expand our advanced system throughout the Asia-Pacific region and studied its usefulness. METHODS: Kyushu University Hospital (Fukuoka, Japan) was linked to 33 medical institutions and meeting venues in China, Taiwan, Thailand, Vietnam, Singapore, and Australia by academic optic fiber network. Digital video signals were directly converted into Internet protocol, and cipher security programs were used to protect patient privacy. RESULTS: Of 49 international surgical teleconferences conducted, 16 were real-time demonstrations of surgery, and 33 involved recorded videos. For 37 events, two stations were connected pier-to pier, and for the remaining 12, multiple stations were connected. The network remained stable, and the time delay between stations was restricted to 0.3-1.0 seconds. Responding to questionnaires, 70.6% of participants rated the image quality as "very good," and 22.5% rated it "good." CONCLUSIONS: We succeeded in establishing a high-quality telesurgical system in a wide area of the Asia Pacific region, and this is the first time high-speed Internet technology has been applied to surgery on such a large scale. Because it is not only of high quality but also economical and easy to set up, we believe this system will promote efficient remote surgical education and active academic exchange worldwide. PMID- 17907988 TI - Technical modifications for laparoscopic cholecystectomy by the left-handed surgeon. AB - There is a complete paucity of literature for left-handed surgeons. Some studies revealed that left-handed surgical residents have lesser operating skills and some surgeons have considered leaving surgery at some point in their career owing to laterality-related frustrations. Most important, whereas minimally invasive surgical techniques have had a profound impact on the treatment of diseased gallbladder, these procedures do not eliminate laterality related to the discomfort of left-handed surgeons. Usually, left-handed surgeons must teach themselves a procedure. They must make modifications and learn some technical tips to make a more comfortable, convenient, and safe intervention. The aim of this study was to describe some modifications made by a left-handed surgeon to perform 52 safe laparoscopic cholecystectomies with standard right-handed instruments in our hospital. These surgical steps could be used in a reproducible way to minimize the recurring difficulties of left-handed learners in a surgical residency program. PMID- 17907989 TI - Laparoscopic spleen-preserving distal pancreatectomy: description of an original posterior approach. AB - We are describing in this paper the original and innovative technique we used to perform a spleen-preserving distal pancreatectomy. With the patient positioned on her right lateral side, we inserted four laparoscopic ports in the left subcostal region to enable an upper view on the spleen and its rear attachments. With this approach, we opened and dissected this plan located between the left kidney and the rear aspect of the spleen and of the pancreas. These structures, once liberated naturally, felt "en-bloc" out of the way because of the patient's lateral positioning and the gravity, exposing the operative field without any artificial retraction. Beyond this greater exposure, this new approach offers many other advantages, such as the easiness to be performed by only two operators and the preservation of the anterior abdominal cavity, the great omentum, the splenic vessels, and the short gastric vessels left untouched. PMID- 17907990 TI - Financial analysis of laparoscopic versus open nephrectomy in the pediatric age group. AB - The authors compared the cost of laparoscopic nephrectomy to open nephrectomy in the pediatric age group. One hundred seventeen consecutive laparoscopic nephrectomies performed by a surgeon with extensive experience with this approach between April 2003 and August 2006 were included. A control group of 24 consecutive open nephrectomies performed by urologists who do not use the laparoscopic approach were also included. Inclusion criteria for surgery were a poor or nonfunctioning kidney related to severe obstructive or refluxing nephropathy and a multicystic dysplastic kidney. The length of operation, length of stay, and disposable equipment used were recorded and the different approaches were compared statistically with an unpaired t test. The mean (standard deviation [SD]) duration of the procedure was 79 minutes (32) in the laparoscopic group and 85 minutes (35) in the control group (P = 0.41). The mean (SD) cost of the disposable instruments used during the operation was pounds sterling274 (160) in the laparoscopic group and pounds sterling20 (5) in the control group (P = 0.0001). The mean (SD) hospital stay was 1 night (0.43) with a mean (SD) cost of pounds sterling677 (291) in the laparoscopic group, and 3 nights (2) with a mean (SD) cost of pounds sterling2031 (1354) in the control group (P = 0.0001). The mean (SD) total cost of the procedure was pounds sterling951 (451) for the laparoscopic group and pounds sterling2051 (1359) for the open one (P = 0.0001). In our experience, the laparoscopic approach in the pediatric age group is 54% less expensive than the open approach. PMID- 17907992 TI - Laparoscopic Nissen fundoplication in an infant with situs inversus. AB - In this paper, we describe the first reported case of a laparoscopic Nissen fundoplication and gastrostomy tube for gastroesophageal reflux disease in a 3 month-old infant with complete abdominal situs inversus. PMID- 17907991 TI - Improved outcomes for laparoscopic appendectomy compared with open appendectomy in the pediatric population. AB - INTRODUCTION: The appendectomy is a common emergent surgical procedure in the pediatric population. The aim of this study was to examine our institution's experience and outcomes in the appendectomy in the pediatric population early in our transition from open surgery to a predominantly laparoscopic approach. METHODS: We retrospectively studied all pediatric patients (age 20 years) that underwent an appendectomy at a tertiary care center over 2 years. The data collected included patient demographics, comorbidities, operative details, outcomes, and complications. RESULTS: Two hundred twenty-three consecutive patients, with a mean age of 9.5 (3.9) years, were included in the study. Forty four laparoscopic and 179 open appendectomies were performed. Two of the laparoscopic cases were converted to open appendectomies. Significant differences were seen between the two groups, with longer operative times (P < 0.0001) and lower estimated blood loss (P = 0.007) in the laparoscopic group. Operative times improved significantly for the laparoscopic group as the surgeons became more experienced (P = 0.03). The laparoscopic group used intravenous pain medication for a shorter time (0.8 vs. 1.9 days; P = 0.0003) and had a shorter postoperative hospital length of stay (2.2 vs. 3.4 days; P = 0.004). The laparoscopic group had fewer wound infections (2.3% vs. 6.2%; P = 0.3), intra-abdominal abscesses (4.5% vs. 5.6%; P = 0.8), and postoperative ileus (0% vs. 2.2%; P = 0.3), although these differences did not reach statistical significance. CONCLUSION: The laparoscopic appendectomy procedure is a safe alternative to open appendectomy in pediatric patients and results in shorter hospital stays with less postoperative pain. PMID- 17907993 TI - Laparoscopic excision of abdominoscrotal hydrocele. AB - Abdominoscrotal hydrocele (ASH) is reported with increasing frequency and is recognized to be responsible for complications not only related to the pressure effect on the contiguous structures, but a wide a variety of conditions, including hemorrhage and malignant transformation. Although there are only two reports in the literature of spontaneous resolution, the actual accepted consensus for treatment is complete excision. The surgical approaches are abdominal, scrotal or combined. There is no report in the literature of a laparoscopic excision of ASH. In this paper, we report on the first case to be treated with this approach and highlight the new advantages and simplicity in using this recommended technique. PMID- 17907994 TI - Combined laparoscopic and thoracoscopic excision of pheochromocytoma and functional paraganglioma. AB - In this paper, we report an 11-year-old girl who underwent a combined laparoscopic adrenalectomy procedure and thoracoscopic excision of a paravertebral mass. To our knowledge, this is the first such report in the English literature. PMID- 17907995 TI - Ligasure vessel sealing system in laparoscopic Palomo varicocele ligation in children and adolescents: much ado about nothing. PMID- 17907997 TI - The future of medical museums: threatened but not extinct. PMID- 17907998 TI - Humanizing medical practice: the role of empathy. PMID- 17907999 TI - Hospital costs of older people in New South Wales in the last year of life. AB - OBJECTIVE: To estimate hospital inpatient costs by age, time to death and cause of death among older people in the last year of life. DESIGN AND SETTING: Cross sectional analytical study of deaths and hospitalisations in New South Wales from linked population databases. PARTICIPANTS: 70,384 people aged 65 years and over who died in 2002 and 2003. MAIN OUTCOME MEASURES: Hospital costs in the year before death. RESULTS: Care of people aged 65 years and over in their last year of life accounted for 8.9% of all hospital inpatient costs. Hospital costs fell with age, with people aged 95 years or over incurring less than half the average costs per person of those who died aged 65-74 years ($7028 versus $17,927). Average inpatient costs increased greatly in the 6 months before death, from $646 per person in the sixth month to $5545 in the last month before death. Cardiovascular diseases (43.1% of deaths) were associated with an average of $11,069 in inpatient costs, while cancer (25.0% of deaths) accounted for $16,853. The highest average costs in the last year of life were for people who died of genitourinary system diseases ($18,948), and the highest average costs in the last month of life were for people who died of injuries ($8913). CONCLUSION: Population ageing is likely to result in a shift of the economic burden of end-of life care from the hospital sector to the long-term care sector, with consequences for the supply, organisation and funding of both sectors. PMID- 17908000 TI - Suicide risk among recently released prisoners in New South Wales, Australia. AB - OBJECTIVE: To determine the risk of suicide and drug overdose death among recently released prisoners. DESIGN, SETTING AND PARTICIPANTS: Retrospective cohort study of 85 203 adult offenders who had spent some time in full-time custody in prisons in New South Wales between 1 January 1988 and 31 December 2002. MAIN OUTCOME MEASURES: Association between time after release and risk of suicide and overdose death. RESULTS: Of 844 suicides (795 men, 49 women), 724 (86%) occurred after release. Men had a higher rate of suicide than women both in prison (129 v 56 per 100,000 person-years) and after release (135 v 82 per 100,000 person-years). The suicide rate in men in the 2 weeks after release was 3.87 (95% CI, 2.26-6.65) times higher than the rate after 6 months. Male prisoners admitted to the prison psychiatric hospital had a threefold higher risk than non-admitted men both in prison and after release. No suicides among women were observed in the 2 weeks after release. No increased risk of suicide was observed among Aboriginal Australians in the first 2 weeks after release. Of 1674 deaths due to overdose, 1627 (97%) occurred after release. Drug-related mortality in men was 9.30 (95% CI, 7.80-11.10) times higher, and in women was 6.42 (95% CI, 3.88-10.62) times higher, in the 2 weeks after release than after 6 months. CONCLUSIONS: Prisoners are at a heightened risk of suicide and overdose death in the immediate post-release period. After 6 months post-release, the suicide rate approaches the rate observed in custody. PMID- 17908001 TI - Postpartum haemorrhage occurrence and recurrence: a population-based study. AB - OBJECTIVE: To determine the risk of occurrence and recurrence of postpartum haemorrhage (excessive bleeding after childbirth) among women having at least two consecutive pregnancies. DESIGN AND SETTING: Population-based study using longitudinally linked hospital discharge and birth records from New South Wales for the period 1 January 1994 to 31 December 2002. PARTICIPANTS: All 125,295 women having at least a first and second pregnancy resulting in a singleton birth at > 400 g or > or = 20 weeks' gestation in the study period. MAIN OUTCOME MEASURES: Risk of occurrence of postpartum haemorrhage (PPH) in any pregnancy, and of recurrence of PPH in subsequent (second and third) pregnancies. RESULTS: 5.8% of women (7327/125,295) had a PPH in their first pregnancy, and 4.5% (5318/117,968) had a first PPH in their second pregnancy. Among the 23,095 women who had three pregnancies in the study period, 4.4% (908/20,839) had a first PPH in their third pregnancy. The risk of recurrence in a second consecutive pregnancy was 14.8% (1082/7327), and in a third consecutive pregnancy (after two previous PPHs) was 21.7% (43/198); even with an intervening pregnancy with no PPH (ie, PPH in the first and third pregnancies only), the risk for the third pregnancy was 10.2% (111/1085). CONCLUSIONS: These consistently elevated risks of recurrence highlight the need for women with a history of PPH to have active management of the third stage of labour and to give birth in a hospital that has onsite blood cross-match facilities. PMID- 17908002 TI - National standard for health assessment of rail safety workers: the first year. AB - OBJECTIVE: To determine the prevalence of health problems in New South Wales train drivers and the impact of the new national health-assessment standard on train drivers' fitness for work. DESIGN, SETTING AND PARTICIPANTS: Retrospective audit of files of all RailCorp train drivers (743) and train driver recruits (283) who were assessed under the new national standard for health assessment of rail safety workers between February 2004 and February 2005. MAIN OUTCOME MEASURES: Smoking status; prevalence of hypertension, heart disease, diabetes and obstructive sleep apnoea; alcohol use disorders; body mass index (BMI); total cholesterol level; fasting blood glucose level; cardiac risk score; fitness status. RESULTS: 25.2% of drivers and 27.9% of recruits were smokers; 43.8% of drivers and 21.9% of recruits were hypertensive; 34.6% of drivers and 31.4% of recruits had high total cholesterol levels (> 5.5 mmol/L). Median BMI values were 29 kg/m(2) (range, 18-59 kg/m(2)) for drivers and 28 kg/m(2) (range, 19-55 kg/m(2)) for recruits. The prevalence of obesity (BMI > or =30.0 kg/m(2)) was higher in both male drivers and recruits compared with the general male population. At initial assessment, 65.1% of drivers and 88.0% of recruits were certified as unconditionally fit for work; 12.4% of drivers and 7.1% of recruits were assessed as temporarily unfit; and 22.5% of drivers and 4.6% of recruits were considered fit subject to review (after periods ranging from 3 to 12 months). Two per cent of drivers and 2.5% of recruits were subsequently deemed to be permanently unfit, the most common reasons being heart conditions, psychiatric disorders, orthopaedic problems, colour vision impairment and sleep apnoea. CONCLUSIONS: Cardiovascular risk factors and cardiovascular disease are the most significant health issues affecting train drivers' fitness for work. With the more stringent health assessment and regular review required by the new standard, most drivers can continue with their duties, with the added benefits of improved personal health and greater safety to the rail network and the public. PMID- 17908003 TI - A spilled gallstone. PMID- 17908004 TI - Disclosure of genetic information to at-risk relatives: recent amendments to the Privacy Act 1988 (Cwlth). AB - The federal Privacy Act 1988 (Cwlth) has recently been amended to permit the disclosure of genetic information to an at-risk relative when there is a serious (although not necessarily imminent) threat to that person's life, health, or safety. This represents a significant exception to the statutory obligations to maintain the privacy of a patient's health information. However, its scope of operation is limited in that it applies only to doctors and other health professionals working in the private sector, and does not cover those working in State public hospitals or for Commonwealth Government agencies. PMID- 17908005 TI - Reforming medical education in the United Kingdom: lessons for Australia and New Zealand. AB - Medical education faces global challenges because of the changing health care needs of an ageing and more demanding society, and the consequent requirement for increased health care workforce capacity and different workforce models. In the United Kingdom, education reform has spanned the medical, nursing and allied health professions, and has introduced new health professions with specific roles within a new, team-based model of comprehensive health care. In medical education, the UK reforms span undergraduate, prevocational, vocational and continuing education, with the aim of providing a framework for faster, more flexible career development that can adapt to future changes in workforce need. While some reforms are controversial, most appear sensible and are supported by most observers. The Modernizing Medical Careers process suffered implementation difficulties in 2007: the national, web-based application scheme for vocational training posts could not cope with such a large process, disrupting both the recruitment of an appropriate workforce for hospitals and the career progression of many recent UK medical graduates. The main problem appears to have been in management of change--too much was attempted too quickly on too large a scale- resulting in a backlash against any significant change. There may be lessons for Australia and New Zealand, which face similar challenges and are considering broadly similar changes. PMID- 17908006 TI - Antifungal agents. AB - The four main classes of antifungal drugs are the polyenes, azoles, allylamines and echinocandins. Clinically useful "older" agents include topical azole formulations (for superficial yeast and dermatophyte infections), first generation triazoles (fluconazole and itraconazole, for a range of superficial and invasive fungal infections), amphotericin B formulations (for a broad range of invasive fungal infections) and terbinafine (for dermatophyte infections). Clinically important "newer" agents include members of the echinocandin class (eg, caspofungin) and second-generation triazoles (eg, voriconazole and posaconazole). Voriconazole and posaconazole have broad-spectrum activity against yeasts and moulds, including Aspergillus species. Posaconazole is the only azole drug with activity against zygomycete fungi. Caspofungin and the other echinocandins are effective in treating Candida and Aspergillus infections. The azoles are relatively safe, but clinicians should be aware of drug-drug interactions and adverse effects, including visual disturbances (with voriconazole), elevations in liver transaminase levels, and skin rashes. Caspofungin has minimal adverse effects. Combination antifungal therapy may be appropriate in selected patients with invasive fungal infections, but is empiric and driven by individual physician practice. PMID- 17908007 TI - Why are community psychiatry services in Australia doing it so hard? AB - Since the National Mental Health Policy was implemented in 1992, dissonance between mental health professionals and the general public on the success of the deinstitutionalization components of the strategy has grown. Many of the premises on which the deinstitutionalization components were based are false, and this has led to many problems in the system. Community psychiatry is not fundamentally flawed, and what has been learned in the past 15 years can be used to build on the foundations that have been laid. Better cooperation between state and federal governments is needed to effect real change. PMID- 17908008 TI - Apparent spontaneous complete regression of a multifocal malignant mesothelioma of the pleura. AB - A 61-year-old woman diagnosed with multifocal, poorly differentiated epithelial mesothelioma in September 2002 went into sustained spontaneous remission within months. She was completely disease-free within 6 months, and remained so 5 years later. This case demonstrates that this tumour may, very rarely, regress spontaneously, with no recurrence for many years. A greater knowledge of the underlying immune mechanisms would aid future management of this and other tumours. PMID- 17908009 TI - Tropical pulmonary eosinophilia: a rare cause of cough in immigrants to Australia. PMID- 17908010 TI - Religion as a competing interest. PMID- 17908011 TI - Supplement unbalanced. PMID- 17908012 TI - Seeking clarification. PMID- 17908013 TI - Gratuitous and without scientific substance. PMID- 17908014 TI - Statements of competing interest notably absent. PMID- 17908016 TI - Religious affiliation and life expectancy at birth. PMID- 17908017 TI - Early intervention in bipolar disorders: opportunities and pitfalls. AB - The early phases of bipolar disorders are difficult to diagnose and have specific treatment issues. The initial polarity of the illness is more commonly depressive, yet in counterpoint, mania is required for diagnosis; consequently, there is often a substantial delay in the initiation of appropriate therapy. There is good evidence that lithium in particular is most effective early in the illness course, and that its efficacy declines after multiple episodes. The notion of neuroprotection reflects this, and furthermore suggests that appropriate therapy may prevent the neurostructural and neurocognitive changes seen in the disorder. Inappropriate therapy may worsen the course of the illness. Patients with a first episode have specific psychosocial needs, and adherence to medication is relatively poor. There is a need for early identification, and to develop treatments and services applicable to the specific needs of this population. PMID- 17908018 TI - Early intervention for depressive disorders in young people: the opportunity and the (lack of) evidence. AB - Young people experiencing their first onset of depression are a group at risk of relapse and recurrence to whom early intervention and prevention efforts should be targeted. Despite the argument for a significant research effort addressing these issues, the evidence regarding optimal intervention strategies for first episodes is lacking. Cognitive behaviour therapy is an effective approach to treatment and relapse prevention among depressed adolescents, and is likely to be an important component of any evidenced-based approach to early intervention. Antidepressants are not recommended as first-line treatment for most first episodes of depression. The role that they may play in patients with severe depression, or those who do not respond to psychological therapies, requires further evaluation. Given the high prevalence of depressive disorders, and the significant burden of disease they represent within our community, early intervention in depressive disorders is a critical research agenda for the future. PMID- 17908019 TI - Prevention and early intervention for borderline personality disorder. AB - Borderline personality disorder (BPD) is a severe mental disorder that is associated with substantial psychosocial impairment and morbidity, disproportionate use of health resources, a high suicide rate, and a reputation for being "untreatable". A diagnosis of BPD in young people has similar reliability, validity and prevalence to BPD in adults, and almost certainly has serious and pervasive negative repercussions over subsequent decades. Current data are inadequate to inform specific universal or selective prevention programs for BPD. However, they do support including BPD prevention as an outcome when evaluating universal and/or selective interventions for a variety of mental health problems and adverse psychosocial outcomes. The strongest data support early intervention for the emerging BPD phenotype. Early intervention programs will need to be realistic in their aims, require change in clinician attitudes and service systems, and must be mindful of the risk of iatrogenic harm. PMID- 17908020 TI - Intervening early to reduce developmentally harmful substance use among youth populations. AB - Early-onset or frequent substance use during adolescence increases the risk of developing mental health problems, as well as a range of other adverse outcomes (eg, alcohol or drug dependence, educational underachievement, health problems, social difficulties) during late adolescence and early adulthood. Increases in rates of risky drinking among young people are particularly concerning, suggesting that an effective, evidence-based alcohol policy and preventive framework needs to be developed. Restricting the supply of licit and illicit substances to adolescents, delaying the age that licit substances can be legally purchased, reducing positive media portrayals of substance use, and banning targeted promotions, should be universal, public prevention priorities. Mass media campaigns need to deliver coherent and credible evidence-based messages to young people, utilising a broad array of dissemination strategies. Clear policy and guidelines for parents regarding appropriate alcohol use for adolescents also need to be developed. Prevention programs should target children and adolescents in families with parents who use drugs, young people who have been suspended from school, or those with mental health problems. Preventive screening and targeted brief interventions can be effectively delivered in a variety of settings by a range of health professionals. PMID- 17908021 TI - Improving mental health literacy as a strategy to facilitate early intervention for mental disorders. AB - Good mental health literacy in young people and their key helpers may lead to better outcomes for those with mental disorders, either by facilitating early help-seeking by young people themselves, or by helping adults to identify early signs of mental disorders and seek help on their behalf. Few interventions to improve mental health literacy of young people and their helpers have been evaluated, and even fewer have been well evaluated. There are four categories of interventions to improve mental health literacy: whole-of-community campaigns; community campaigns aimed at a youth audience; school-based interventions teaching help-seeking skills, mental health literacy, or resilience; and programs training individuals to better intervene in a mental health crisis. The effectiveness of future interventions could be enhanced by using specific health promotion models to guide their development. PMID- 17908022 TI - Reach Out! Innovation in service delivery. AB - With 75% of mental illness beginning before 25 years of age, it is essential that we improve young people's capacity to manage adversity and increase their opportunities for accessing professional help. With its unique ability to connect people to information and to other people, the Internet offers opportunities to engage the 70% of young people with mental health problems who currently are not seeking professional help. Reach Out! is a national Internet-based mental health service for young people. It has been accessed by over 6 million users since its launch in 1998. Reach Out! plays a role in the prevention of mental health problems by: facilitating help-seeking and connecting young people with services, such as general practitioners, and allied and mental health professionals in their local communities; and providing opportunities for all young people to develop the skills and capacity to better understand mental health difficulties and manage adversity, thereby complementing traditional support. PMID- 17908023 TI - When and how do young people seek professional help for mental health problems? AB - Despite the high prevalence of mental health problems and disorders that develop in adolescence and early adulthood, young people tend to not seek professional help. Young men and young people from Indigenous and ethnic minority groups tend to be those most reluctant to seek help. Young people are more inclined to seek help for mental health problems if they: have some knowledge about mental health issues and sources of help; feel emotionally competent to express their feelings; and have established and trusted relationships with potential help providers. Young people are less likely to seek help if they: are experiencing suicidal thoughts and depressive symptoms; hold negative attitudes toward seeking help or have had negative past experiences with sources of help; or hold beliefs that they should be able to sort out their own mental health problems on their own. Young people may seek help through talking to their family and friends, with family being more important for younger adolescents, and friends and partners becoming more influential later on. The professionals most likely to act as gatekeepers to mental health services for young people are school counsellors, general practitioners, and youth workers. Increasingly, Internet-based information and interventions are being used to engage young people in the help seeking process. PMID- 17908024 TI - Clinical staging: a heuristic model for psychiatry and youth mental health. AB - Diagnosis in psychiatry continues to struggle to fulfil its key purposes, namely to guide treatment and to predict outcome. A clinical staging model, widely used in clinical medicine, could improve the utility of diagnosis in psychiatry, especially in young people with emerging disorders. Clinical staging has immediate potential to improve the logic and timing of interventions in psychiatry, as it does in many complex and potentially serious medical disorders. Interventions could be evaluated in terms of their ability to prevent or delay progression from earlier to later stages of a disorder, and selected by consumers and clinicians on the basis of clear-cut risk-benefit criteria. This would ensure that, as treatments are offered earlier, they remain safe, acceptable and affordable, and potentially more effective. Biological variables and a range of candidate risk and protective factors could be studied within and across stages, and their role, specificity and centrality in risk, onset and progression of disorders clarified. In this way, a clinicopathological framework could be progressively constructed. Clinical staging, with restructuring across and within diagnostic boundaries and explicit operational criteria for extent and progression of disorder, should be actively explored in psychiatry as a heuristic strategy for developing and evaluating earlier, safer, and more effective clinical interventions, and for clarifying the biological basis of psychiatric disorders. Young people with emerging mental and substance use disorders could be the main beneficiaries. PMID- 17908025 TI - PACE: a specialised service for young people at risk of psychotic disorders. AB - Intervention in the prodromal phase of schizophrenia and related psychotic disorders may prevent or delay the onset of these disorders, or reduce the severity of the psychosis. Identifying the schizophrenia prodrome is difficult, however, because of its non-specific symptoms and the wide symptom variability between individuals. Over the past 15 years, we have investigated the schizophrenia prodrome and developed criteria for detecting people suspected of experiencing a prodromal phase (ie, they are thought to be at imminent risk of onset of a psychotic disorder). About 35% of those meeting our criteria have developed a psychotic disorder within 12 months. We have established a clinical service, the PACE (Personal Assessment and Crisis Evaluation) Clinic, for people with suspected incipient psychosis, and trialled interventions aimed at preventing or delaying the onset of psychotic disorders. Our results and studies in other countries seem to indicate that psychological and psychosocial interventions, either alone or in combination with pharmacotherapy, may be effective in at least delaying, if not preventing, the onset of a psychotic disorder. PMID- 17908026 TI - Responding to experiences of young people with common mental health problems attending Australian general practice. AB - The development of evidence-based ("collaborative care") mental health services in primary care for young people with anxiety, depression and alcohol or other substance misuse is a major challenge. Data from two clinical audits of selected Australian general practices (1998-1999 and 2000-2002) were analysed to explore actual experiences of care among people aged 16-25 years. Syndromal (1998-1999: 31.0% [n = 1849/5957]; 2000-2002: 37.8% [n = 148/392]) and subsyndromal (1998 1999: 27.4% [n = 1635/5957]; 2000-2002: 29.1% [114/392]) mental disorders are very common among young people presenting to general practitioners. However, a mental health diagnosis (1998-1999: 42.6% [n = 740/1736]; 2000-2002: 52.0% [n = 77/148]) or provision of formal treatment (1998-1999: 36.1% [n = 600/1661]; 2000 2002: 51.7% [n = 74/143]) occurs in only about half of the patients with syndromal conditions. While some active treatment was received by 19.4% (1998 1999 [n = 1018/5236]) and 35.9% (2000-2002 [n = 133/370]) of the young people, respectively, the most commonly reported interventions were non-pharmacological alone (1998-1999: 13.1% [n = 687/5236]; 2000-2002: 22.4% [n = 83/370]) or non pharmacological and pharmacological combinations (1998-1999: 4.1% [n = 214/5236]; 2000-2002: 10.3% [n = 38/370]). Only rarely is pharmacological treatment alone provided (1998-1999: 2.2% [n = 117/5236]; 2000-2002: 3.2% [n = 12/370]). New systems of primary care for young people need to be based on proven collaborative care models and encourage presentations for care, increase detection rates, and promote access to information and effective e-health services. Improved access to specific psychological treatments should remain a priority. PMID- 17908028 TI - The specialist youth mental health model: strengthening the weakest link in the public mental health system. AB - Despite mental disorders being the dominant health issue confronting young people, youth mental health is yet to be recognised as a discrete, unified program area; responsibility for young people's mental health is currently split across multiple levels of government. Public specialist mental health services have followed a paediatric-adult split in service delivery, mirroring general and acute health care. The pattern of peak onset and the burden of mental disorders in young people means that the maximum weakness and discontinuity in the system occurs just when it should be at its strongest. Young people need youth-friendly services that recognise and respond to their special cultural and developmental needs. At the primary and community level, headspace: the National Youth Mental Health Foundation, is a national response to this and aims to provide better access, engagement and enhanced multidisciplinary care for young people across Australia. The specialist mental health service level should be complemented by youth-specific specialist mental health services for young people, aged 12-25 years, which would strengthen the existing system with a better targeted stream of care, providing access to integrated mental health, substance use, and vocational-recovery services. Alternative approaches to creating this capacity should be urgently developed and evaluated, and sustained reform informed by evidence as well as values. PMID- 17908029 TI - Principles of youth participation in mental health services. AB - Young people with mental illness face many barriers in accessing care and often have different needs to those of adult consumers. Young people's participation in mental health services is one way of addressing quality and access issues, through receiving feedback and implementing youth-driven and youth-friendly strategies. headspace, the National Youth Mental Health Foundation, established in July 2006, highlights the mental health care sector's commitment to young people. Existing youth participation programs provide examples of what can be achieved at national and local levels and with varying levels of financial and other support. These include: Ybblue, the youth program of beyondblue; Reach Out!, a web-based service; Headroom, providing health promotion and a website; and Platform Team (ORYGEN Youth Health), comprising current and past clients who advise the service and provide peer support. Current practice in youth participation in mental health services involves a variety of methods, such as ensuring information and education is appropriate for a youth audience, and participating in peer-support programs and staff selection panels. Challenges in the future development of youth participation in mental health services include avoiding tokenism, acknowledging that young people are not a uniform group, translating national strategies into local improvements in services, and gaining the support and cooperation of health care workers in genuine participation. PMID- 17908030 TI - Minimising collateral damage: family peer support and other strategies. AB - The impact on family members of mental illness in a young person is intensely distressing. Symptoms that they cannot understand, and the stigma surrounding mental illness may lead to families feeling isolated in their distress. Family carers are reassured by talking with other families who have experienced the same or similar situations. The "Families Helping Families" program at ORYGEN Youth Health trains family peer-support workers, who are employed to provide information and support to families new to the service. Medical practitioners need to appreciate the importance of ensuring that families receive information and emotional support to help them cope effectively. Difficulties in helping family carers, often around perceived confidentiality restraints, need to be overcome so that collateral damage--family breakdown, persistence of symptoms, and behavioural maladjustments--can be reduced. Families who are supported can become advocates for improvements to mental health services for young people. PMID- 17908031 TI - Tips and techniques for engaging and managing the reluctant, resistant or hostile young person. AB - Creating a collaborative doctor-patient relationship is the bedrock upon which effective treatments are delivered. The interaction between normal developmental changes and psychopathology can present particular challenges to clinicians attempting to assess and treat young people. Assuming an attitude in which young people are seen to be doing their best, rather than being deliberately difficult or manipulative, can help clinicians avoid a controlling or punitive relationship and can facilitate collaborative problem solving. Stigma, denial and avoidance, ambivalence, hopelessness and coercion are potential threats to engagement and must be addressed specifically. Challenging patients, such as the reluctant, resistant, aggressive, self-harming or intoxicated patient require specific management strategies that can be learned. PMID- 17908032 TI - headspace: Australia's National Youth Mental Health Foundation--where young minds come first. AB - headspace, Australia's national youth mental health initiative, was created in 2006 in response to the recognition that the existing health system needed to be much more accessible and effective for young people with mental and substance use disorders. With funding of more than $54 million from the Australian Government, a carefully constructed and selected system of 30 "communities of youth services", or integrated service hubs and networks, across the nation is being established, supported by programs for community awareness, workforce training and evidence-based resource material. headspace aims to improve access, and service cohesion and quality, and ultimately health and social outcomes, for young people aged 12-25 years experiencing mental illness and related substance use problems. Within the Council of Australian Governments framework, this will require synergistic planning with, and co-investment on behalf of, state and territory governments, as well as the support and involvement of local communities and the wider Australian society. PMID- 17908033 TI - Early intervention in psychotic disorders: detection and treatment of the first episode and the critical early stages. AB - The two main goals of early intervention in psychotic disorders are to reduce the period of time between the onset of psychosis and the commencement of effective treatment, and to provide consistent and comprehensive care during the critical early years of illness. Effective care during the critical early years involves proactive engagement and initiation of drug and psychosocial treatments, aiming for maximal symptomatic and functional recovery and the prevention of relapse. Over the past 15 years, an increasing number of specialised or streamed treatment delivery systems for early psychosis have been established around the world. There is now evidence that these services can reduce the duration of untreated psychosis and produce better symptomatic and functional recovery. In addition, they are more cost-effective than standard models of mental health care for these patients. Fully fledged, specialised early intervention services should be established, with full integration with local communities, as well as enhanced primary care systems focused on young people. PMID- 17908034 TI - Advances in Alzheimer therapy: development of innovative new strategies. PMID- 17908035 TI - Cholinotrophic molecular substrates of mild cognitive impairment in the elderly. AB - Cholinergic nucleus basalis (NB) neurons provide the major cholinergic innervation to the cortical mantle, are selectively vulnerable in late stage Alzheimer's disease (AD) and require the neurotrophin, nerve growth factor (NGF) and its receptors (TrkA and p75(NTR)), for their survival. The molecular events underlying the demise of these neurons in AD were investigated using tissue harvested from participants in a longitudinal clinical pathological study of aging and AD who agreed to an annual clinical evaluation providing a categorization of no cognitive impairment (NCI), mild cognitive impairment (MCI) or AD and postmortem brain donation. Although the number of choline acetyltransferase (ChAT)-positive neurons was unchanged, TrkA and p75(NTR) receptor-containing neurons, which co-localize with ChAT, were significantly reduced in the NB of subjects with MCI and AD compared to those with NCI. These observations indicate a phenotypic down-regulation rather than frank NB neuronal degeneration in MCI. Expression profiling of single cholinergic NB neurons revealed TrkA but not p75(NTR) mRNA is reduced in MCI, suggesting that decreased neurotrophin responsiveness may be an early biomarker for AD. The NGF precursor molecule, proNGF, is increased in the cortex in MCI and AD. Since proNGF accumulates in the presence of reduced cortical TrkA and sustained levels of p75(NTR), a shift in the balance between cell survival and death molecules may occur in prodromal AD. Coincident with these phenomena, brain derived neurotrophic factor (BDNF) and its precursor molecule, proBDNF, are reduced in the MCI cortex, potentially depriving CBF neurons of additional trophic factor support. Moreover, there is a shift in the ratio of 3 repeat tau to 4 repeat tau gene expression, whereas total tau message is stable in NB neurons during the disease process. These data suggest there is a shift in cholinotrophic molecular events in MCI and early AD which may lead to cell dysfunction and eventual cell death over the course of the disease. These findings support the concept that from a neurotrophic pathobiologic perspective, MCI is already early AD. PMID- 17908036 TI - NGF-cholinergic dependency in brain aging, MCI and Alzheimer's disease. AB - Forebrain cholinergic neurons are highly dependent on nerve growth factor (NGF) for phenotype maintenance. We have established that in addition to "target derived" NGF neurotrophic stimulation, cholinergic neurons also respond dose dependently, to intra-parenchymal NGF administration in the somato-dendritic region of the nucleus Basalis, thus illustrating the potential of alternative reparative therapies which would by-pass the undesirable effects of diffuse neurotrophin application. Moreover, our lab has also observed that the steady state number of cortical cholinergic synapses is dependent on continuous NGF supply, as anti-NGF monoclonal antibodies and TrkA receptor antagonists deplete pre-existing cholinergic bouton numbers. Furthermore, the application of either NGF or TrkA NGF-mimetic agonists successfully rescues the age-dependent loss of cortical cholinergic boutons in aged-impaired rats. The vulnerability of the cortical cholinergic system has also been demonstrated in transgenic animal models of the Alzheimer's disease (AD) amyloid pathology. It is of interest to note however, that an up-regulation of cholinergic presynaptic boutons has been observed in certain transgenic mouse models prior to plaque formation. This observation is similar to the visibly increased immunoreactivity of cortical and hippocampal choline acetyltransferase (ChAT) fibers in patients with Mild Cognitive Impairment (MCI). A series of ex-vivo experiments conducted by our group have demonstrated that contrary to popular belief, proNGF, as opposed to mature NGF, is released from the cerebral cortex in an activity-dependent manner. In addition, proNGF appears to be released with a series of pro-enzymes and enzymes, which are involved in its subsequent maturation to NGF and degradation in the extracellular space. Given that proNGF is known to be upregulated in AD patients a dysregulation in the maturation or degradation of mature NGF might explain the preferential vulnerability of the cholinergic system in the AD pathology. PMID- 17908037 TI - Amyloid beta protein as a marker or risk factor of Alzheimer's disease. AB - Alzheimer disease (AD) is a neurodegenerative disease that affects cognition, behavior and function. The etiology of the disease is unknown, however, the Primary Risk Factors for AD are aging, and family history. Neurofibrillary tangles (NFT) and amyloid-bearing neuritic plaques in the limbic and cerebral cortices are the characteristic neuropathologic lesions in brains of patients with AD. The NFT is mainly composed of hyprephosphorylated tau, whereas the major component of the neuritic plaques is the amyloid beta (Abeta) protein. The clinical diagnosis of probable AD is based on history, physical examination, neuropsychological testing, laboratory studies and neuroimaging techniques. However, there is no specific laboratory marker to support the diagnosis of definite AD or monitoring the progression of the disease. Several biochemical markers related to neuropathology have been identified in cerebrospinal fluid (CSF). We describe the studies of CSF or blood levels of amyloid beta protein in patients with AD and age-matched nondemented controls. Due to the heterogeneity and complex nature of the disease, it is highly unlikely that that a single marker specific for AD will be found. PMID- 17908038 TI - Computerized methods in the assessment and prediction of dementia. AB - Computerized administration of neuropsychological tests can be an objective, sensitive and efficient way to screen for and monitor cognitive changes in the elderly. However, current computer software still suffers from limitations in both the administration of those tests and the interpretation of their results, which might severely hamper their usability. In this paper qualitative aspects of current methods and their use in the prediction of dementia are discussed, guidelines for correct design and usage of computerized methods are suggested and a solution that overcomes several of the methodological limitations is proposed. PMID- 17908039 TI - Practical issues in stem cell therapy for Alzheimer's disease. AB - We have demonstrated that aged animals show significant improvements in cognitive function and neurogenesis after brain transplantation of human neural stem cells or of human adult mesenchymal stem cells that have been dedifferentiated by transfection of the embryonic stem cell gene. We have also demonstrated that peripheral administration of a pyrimidine derivative increased cognition, endogenous brain stem cell proliferation and neurogenesis. These results indicate a bright future for stem cell therapies in Alzheimer's disease (AD). Before this is realized, however, we need to consider the affect of AD pathology on stem cell biology to establish an effective stem cell therapy for this disease. Although amyloid-beta (Abeta) deposition is a hallmark of AD, an absence of a phenotype in the beta-amyloid precursor protein (APP) knockout mouse, might lead one to underestimate the potential physiological functions of APP and suggest that it is unessential or can be compensated for. We have found, however, that APP is needed for differentiation of neural stem cells (NSCs) in vitro, and that NSCs transplanted into a APP-knockout mouse did not migrate or differentiate -- indicating that APP plays an important role in differentiation or migration process of NSCs in the brain. Then again, treatment with high a concentration of APP or its over-expression increased glial differentiation of NSCs. Human NSCs transplanted into APP-transgenic mouse brain exhibited less neurogenesis and active gliosis around the plaque like formations. Treatment of such animals with the compound, (+)-phenserine, that is known to reduce APP protein levels, increased neurogenesis and suppressed gliosis. These results suggest APP levels can regulate NSC biology in the adult brain, that altered APP metabolism in Down syndrome or AD may have implications for the pathophysiology of these diseases, and that a combination of stem cell therapy and regulation of APP levels could provide a treatment strategy for these disorders. PMID- 17908040 TI - TNF-alpha inhibition as a treatment strategy for neurodegenerative disorders: new drug candidates and targets. AB - As the average ages of North Americans and Europeans continue to rise; similarly the incidence of "old age" associated illnesses likewise increases. Most notably among these ailments are conditions linked to dementia-related neurodegenerative disorders, such as Alzheimer's disease (AD), Parkinson's disease (PD) and stroke. While in the early stages, these conditions are associated with cellular dysfunction in distinctly different brain regions, thus affecting different neuronal cell types; it is most likely that the final stages share similar cellular and molecular processes leading to neuronal death and ultimately overt clinical symptoms. In this regard, different environmental and genetic triggers ranging from head trauma to protein mutations and toxicological exposure may instigate a cascade of intracellular events that ultimately lead to neuronal death. One strong candidate trigger protein, and thus a potential target for therapeutic manipulation is the potent pro-inflammatory / pro-apoptotic cytokine, tumor necrosis factor-alpha (TNF-alpha). TNF-alpha is secreted by the brain resident marcophage (the microglial cell) in response to various stimuli. It has been demonstrated to play a major role in central nervous system (CNS) neuroinflammation-mediated cell death in AD, PD and amyotrophic lateral sclerosis (ALS) as well as several other CNS complications. Recently, agents that modulate the levels of circulating peripheral TNF-alpha protein have been shown to be worthwhile therapeutic agents with the use of Enbrel (Etanercept) and Remicade (Infliximab), both of which display beneficial properties against rheumatoid arthritis and other peripheral inflammatory diseases. Unfortunately, these agents are largely unable to penetrate the blood-brain barrier, which severely limits their use in the setting of neuroinflammation in the CNS. However, thalidomide, a small molecule drug, can inhibit TNF-alpha protein synthesis and, unlike larger molecules, is readily capable of crossing the blood-brain barrier. Thus thalidomide and its analogs are excellent candidate agents for use in determining the potential value of anti-TNF-alpha therapies in a variety of diseases underpinned by inflammation within the nervous system. Consequently, we have chosen to discuss the relevance of unregulated TNF-alpha expression in illnesses of the CNS and, to an extent, the peripheral nervous system. Additionally, we consider the utilization of thalidomide-derived agents as anti-TNF-alpha therapeutics in the setting of neuroinflammation. PMID- 17908041 TI - East meets West in the search for Alzheimer's therapeutics - novel dimeric inhibitors from tacrine and huperzine A. AB - Alzheimer's disease (AD) is linked to cholinergic deficiency and the overactivation of glutamate receptors. The acetylcholinesterase (AChE) inhibition treatment approach has produced the most encouraging results in clinical practice, and memantine, a moderate antagonist of N-methyl-D-aspartate (NMDA) receptors, has been approved for treating AD. However, AChE inhibitors have limited success as they only improve memory in mild dementia but cannot stop the process of neurodegeneration; while memantine possesses neuroprotective effects only with a little ability in memory enhancement. There has been a major rush among neuroscience research institutions and pharmaceutical firms worldwide to search for safer and more effective therapeutic agents for AD. The novel dimers, derived from tacrine and the fragment of huperzine A (HA'), have been demonstrated to be potent and selective reversible inhibitors of AChE. Bis(7) tacrine, bis(12)-hupyridone (E12E) and HA'(10)-tacrine, are representatives of three series of novel dimers. According to the preclinical studies, these compounds have been shown to have low toxicity and high efficacy for improving cognitive deficits in several animal models. More interestingly, bis(7)-tacrine, similar to memantine, prevents glutamate-induced neurotoxicity by moderately blocking glutamate receptor NMDA subtype. Furthermore, bis(7)-tacrine, as well as E12E, possesses multiple neuroprotective effects in vitro and in vivo. Taking together, these dimeric AChE inhibitors, especially bis(7)-tacrine, E12E and HA'(10)-tacrine, may provide beneficial effects in AD and other neurodegenerative diseases. PMID- 17908042 TI - N-phenylamine derivatives as aggregation inhibitors in cell models of tauopathy. AB - Cell models of tauopathy were generated in order to study mechanisms of neurodegeneration involving abnormal changes of tau. They are based on neuroblastoma cell lines (N2a) that inducibly express different forms of the repeat domain of tau (tau(RD)), e.g. the 4-repeat domain of tau with the wild type sequence, the repeat domain with the DeltaK280 mutation ("pro-aggregation mutant"), or the repeat domain with DeltaK280 and two proline point mutations ("anti-aggregation mutant"). The data indicate that the aggregation of tau(RD) is toxic, and that aggregation and toxicity can be prevented by low molecular weight compounds, notably compounds based on the N-phenylamine core. Thus the cell models are suitable for developing aggregation inhibitor drugs. PMID- 17908043 TI - Neurorescue activity, APP regulation and amyloid-beta peptide reduction by novel multi-functional brain permeable iron- chelating- antioxidants, M-30 and green tea polyphenol, EGCG. AB - Accumulation of iron at sites where neurons degenerate in Parkinson's disease (PD) and Alzheimer's disease (AD) is thought to have a major role in oxidative stress induced process of neurodegeneration. The novel non-toxic lipophilic brain permeable iron chelators, VK-28 (5- [4- (2- hydroxyethyl) piperazine-1-ylmethyl] quinoline- 8- ol) and its multi-functional derivative, M-30 (5-[N-methyl-N propargylaminomethyl]-8-hydroxyquinoline), as well as the main polyphenol constituent of green tea (-)-epigallocatechin-3-gallate (EGCG), which possesses iron metal chelating, radical scavenging and neuroprotective properties, offer potential therapeutic benefits for these diseases. M-30 and EGCG decreased apoptosis of human SH-SY5Y neuroblastoma cells in a neurorescue, serum deprivation model, via multiple protection mechanisms including: reduction of the pro-apoptotic proteins, Bad and Bax, reduction of apoptosis-associated Ser139 phosphorylated H2A.X and inhibition of the cleavage and activation of caspase-3. M-30 and EGCG also promoted morphological changes, resulting in axonal growth associated protein-43 (GAP-43) implicating neuronal differentiation. Both compounds significantly reduced the levels of cellular holo-amyloid precursor protein (APP) in SH-SY5Y cells. The ability of theses novel iron chelators and EGCG to regulate APP are in line with the presence of an iron-responsive element (IRE) in the 5'-untranslated region (5'UTR) of APP. Also, EGCG reduced the levels of toxic amyloid-beta peptides in CHO cells over-expressing the APP "Swedish" mutation. The diverse molecular mechanisms and cell signaling pathways participating in the neuroprotective/neurorescue and APP regulation/processing actions of M-30 and EGCG, make these multifunctional compounds potential neuroprotective drugs for the treatment of neurodegenerative diseases, such as PD, AD, Huntington's disease and amyotrophic lateral sclerosis. PMID- 17908044 TI - Alpha-secretase as a therapeutic target. AB - In the non-amyloidogenic pathway the alpha-secretase cleaves the amyloid precursor protein (APP) within the sequence of Abeta-peptides and precludes their formation. In addition, alpha-secretase cleavage releases an N-terminal extracellular domain with neurotrophic and neuroprotective properties. The disintegrin metalloproteinase ADAM10 has been shown to act as alpha-secretase in vivo, to prevent amyloid plaque formation and hippocampal defects in an Alzheimer disease mouse model. An increase in alpha-secretase activity therefore is an attractive strategy for treatment of AD and may be achieved by modulating selective signalling pathways. Functional characterization of the human ADAM10 promoter showed that it contains several binding elements for transcription factors which are regulated by extracellular ligands. Retinoic acid (RA) was identified as an inducer of human ADAM10 promoter activity. In human neuroblastoma cell lines RA treatment upregulated the expression of both the alpha-secretase ADAM10 and its substrates APP and the related APP-like-protein 2 (APLP2), and led to an enhanced secretion of their extracellular domains. Furthermore, G protein-coupled receptors (GPCRs) localized in brain areas affected by AD were investigated. Activation of the PAC1 receptor by the neuropeptide PACAP was identified as an approach for upregulation of the alpha secretase pathway. PMID- 17908045 TI - Memapsin 2 (beta-secretase) inhibitor drug, between fantasy and reality. AB - A major strategy for the development of a disease-modifying therapy against Alzheimer's disease is pharmacological intervention designed to reduce levels of beta-amyloid in the brain. Among various ways of reducing beta-amyloid production, the inhibition of beta-secretase (memapsin 2, BACE) is particularly attractive. Not only does beta-secretase initiates the amyloid cascade, it also is an aspartic protease, a class of proteases for which successful inhibitor drugs have been developed to treat AIDS patients. Extensive efforts in research and development of a beta-secretase inhibitor drug have taken place in many laboratories during the past few years. However, no drug candidate is currently in clinical trials. In spite of the lack of obvious success, much progress has been made to incorporate the drug-like properties in the evolution of better inhibitors. The inhibitors from more recent generations are indeed similar in characteristics to other protease inhibitor drugs. This progress permits optimism that development of clinical candidates of beta-secretase inhibitor drugs is a realistic goal. PMID- 17908046 TI - The gamma/epsilon-secretase-derived APP intracellular domain fragments regulate p53. AB - Amyloid beta-peptide (Abeta), which plays a central role in Alzheimer Disease, is generated by presenilin-dependent and presenilin-independent gamma-secretase cleavages of beta-amyloid precursor protein (betaAPP). We report that the presenilins (PS1 and PS2) also regulate p53-associated cell death. Thus, we established that PS deficiency, catalytically inactive PS mutants, gamma secretase inhibitors and betaAPP or APLP2 depletion reduced the expression and activity of p53, and lowered the transactivation of its promoter and mRNA levels. p53 expression was also reduced in the brains or betaAPP-deficient mice or in brains where both PS had been invalidated by double conditional knock out. AICDC59 and AICDC50, the gamma- and epsilon-secretase-derived C-terminal fragments of betaAPP, respectively, trigger the activation of caspase-3, p53 dependent cell death, and increase p53 activity and mRNA. Finally, HEK293 cells expressing PS1 harboring familial AD (FAD) mutations or FAD-affected brains, all display enhanced p53 activity and p53 expression. Our studies demonstrate that AICDs control p53 at a transcriptional level, in vitro and in vivo and unravel a still unknown function for presenilins. PMID- 17908047 TI - Novel Abeta immunogens: is shorter better? AB - Active and passive Abeta immunotherapy in Alzheimer's disease (AD)-like mouse models lowers cerebral amyloid-beta protein (Abeta) levels, especially if given early in the disease process, and improves cognitive deficits. In 2002, a Phase IIa clinical trial was halted due to meningoencephalitis in approximately 6% of the AD patients. It is hypothesized that the immunogen, full-length Abeta1-42, may have led to an autoimmune response. Currently, we are developing novel Abeta peptide immunogens for active immunization in amyloid precursor protein transgenic mice (APP Tg) to target Abeta B cell epitopes (within Abeta1-15) and avoid Abeta-specific T cell epitopes (Abeta16-42) so as to generate a safe and effective AD vaccine. Intranasal immunization with dendrimeric Abeta1-15 (16 copies of Abeta1-15 on a lysine core) or a tandem repeat of Abeta1-15 joined by 2 lysines and conjugated to an RGD motif with a mutated form of an E. coli-derived adjuvant generated robust Abeta titers in both wildtype and APP Tg mice. The Abeta antibodies recognized a B cell epitope within Abeta1-7, were mostly T helper 2 associated immunoglobulin isotypes, bound human AD and APP Tg plaques, and detected Abeta oligomers. Splenic T cells reacted to the immunogens but not full-length Abeta. Six months of intranasal immunization (from 6-to-12 months of age) of J20 mice with each immunogen lowered insoluble Abeta42 by 50%, reduced plaque burden and gliosis, and increased Abeta in plasma. Interestingly, Abeta antibody generation was influenced by route of immunization. Transcutaneous immunization with dbeta1-15, but not full-length Abeta, led to high Abeta titers. In summary, our short Abeta immunogens induced robust titers of predominantly Th2 antibodies that were able to clear cerebral Abeta in the absence of Abeta specific T cell reactivity, indicating the potential for a safer vaccine. We remain optimistic about the potential of such a vaccine for prevention and treatment of AD. PMID- 17908048 TI - A novel immunotherapy for Alzheimer's disease: antibodies against the beta secretase cleavage site of APP. AB - One of the main neuropathological lesions observed at brain autopsy of Alzheimer's disease (AD) patients are the extracellular senile plaques mainly composed of amyloid-beta (Abeta) peptides. Abeta is generated by proteolytic processing of amyloid precursor protein (APP) via beta and gamma-secretases. The beta-secretase APP cleaving enzyme 1 (BACE1) has become a target of intense research aimed at blocking the enzyme activity. Recent studies showed that BACE1 is involved in processing other non-APP substrates, and that other proteases are involved in APP processing. We have recently established a novel approach to inhibit Abeta production via antibodies against the beta-secretase cleavage site of APP. These antibodies bind wild type and Swedish mutated APP expressed in transgenic mice brain tissues. The isolated antibodies do not bind any form of Abeta peptides. Antibody up-take experiments, using Chinese hamster ovary cells expressing wild-type APP, suggest that antibody internalization and trafficking are mediated via the endocytic pathway. Administration of antibodies to the cells growing media resulted in a considerable decrease in intracellular Abeta levels, as well as in the levels of the corresponding C-terminal fragment (C99). The relevance of intra-neuronal accumulation of mainly Abeta42 as an early event in AD pathogenesis suggests that this approach may be applicable as a novel therapeutic strategy in AD treatment. PMID- 17908049 TI - Is alpha-synuclein pathology a target for treatment of neurodegenerative disorders? AB - Alpha-synuclein is the main constituent of intra-neuronal Lewy bodies, which are characteristic of Parkinson's disease, but aggregates are also found as axonal inclusions. Alpha-synuclein pathology is found together with beta-amyloid plaques and neurofibrillary tangles in Alzheimer's disease and other neurodegenerative disorders. In spite of the fact that the biological function of this synaptic protein is not known so far, there is an increasing body of evidence indicating an interaction with amyloid peptides, but also with tau-hyperphosphorylation. A high proportion of alpha-synuclein purified from Lewy bodies is phosphorylated on Ser129. There are still different opinions about the toxicity of the alpha synuclein aggregates. Alpha-synuclein seems to influence different intracellular signaling pathways which are in direct relation to defense mechanisms against reactive oxygen species or apoptosis. It is obvious that overproduction of alpha synuclein, but also different mutations, are inducing the formation of aggregates. Because of the possible link to neurodegeneration, different attempts have been made to counteract alpha-synuclein aggregation. An interesting approach is utilizing beta-synuclein, a biological factor, with an aminoacid sequence closely resembling that of alpha-synuclein. Proof of concept studies indicated that overexpression of beta-synuclein is able to counteract alpha-synuclein aggregation in a transgenic animal model, while also ameliorating functional deficits. As an alternative approach, the use of low molecular beta-synuclein N terminal peptide derivatives has been considered. Several of these structures displayed clear neuroprotective activities in tissue culture models of neurodegeneration, including beta-amyloid toxicity. Therefore it has been speculated that these compounds might have a broad therapeutic efficacy in different neurodegenerative disorders. A proof of concept study in hAPP transgenic animals resulted in a highly significant decrease in beta-amyloid plaque load, an increase in soluble beta-amyloid peptides and a decrease in insoluble forms. There was also significant improvement of cognitive deficits in this APP transgenic mouse model following intranasal but also peripheral treatment with three of these compounds. From this study it is concluded that the observed effects of the peptides derived from beta-synuclein N-terminus are depending on both, a direct interaction with aggregation of proteins, but also with stimulation of anti-apoptotic and anti-oxidative intracellular signaling pathways. PMID- 17908050 TI - Lessons from Darwin: 21st century designs for clinical trials. AB - What are the resources needed by clinical pharmacology to test drugs in ways that model how the practitioner achieves optimal effectiveness and safety with each patient? I describe the applications of test-retest standard error of measurement, clinical decision rules, means or other statistical summaries of observations, clinical trial designs that use each patient as her own control, and methods to control observer and site variance as steps for developing a CT tested model for optimal clinical uses of an Alzheimer's drug by a practitioner. Many investigators and clinicians have been concerned with clinical judgments being scientifically uncontrolled and unsystematic. The methods I describe demonstrate how clinical trials can be used to overcome these limitations in current patient care. "Darwin showed that one simply could not understand evolution as long as one accepted essentialism. Species and populations are not types, they are not essentialistically defined classes, but rather are biopopulations composed of genetically unique individuals" E. Mayr. PMID- 17908051 TI - Drug trials in dementia: challenging ethical dilemmas. AB - Advances in the treatment of demented individuals is critically dependent upon experimental administration of new drugs to such people, who, by definition, frequently cannot provide informed consent. Ethical problems associated with studies on demented individuals are therefore of great importance. While there is some similarity to other groups (children, psychotic individuals and patients in coma) there also exist several differences. Obtaining an informed consent from a dementing individual is always problematic. Advance directives are helpful to caregivers and patients should be encouraged, at early stages of the disease, to provide them. Participation in drug studies carries inherent benefits to patients, but at the same time exposes them to risks and discomforts which should be monitored and reviewed more intensively than in studies on cognitively intact individuals. The vulnerability of demented people and their dependence requires special attention by the institutional review board (IRB), unique items to be included in the study protocols and consent forms, etc. Representatives of patients' advocacy groups can make important contributions to the IRB, to serve the best interests of the patient and prevent exploitation by the industry as well as by researchers, and honoring the autonomy of the patients. It would be helpful and justified to enable subjects to continue in the study in an open label design in this situation, once they sign a suitable informed consent. A no fault insurance could be provided to the patient in this situation. PMID- 17908052 TI - Alzhemed: a potential treatment for Alzheimer's disease. AB - As a potential disease-modifying treatment for AD, Alzhemed (tramiprosate) is a compound that binds to soluble amyloid-beta peptide (Abeta) and inhibits the formation of neurotoxic aggregates that lead to amyloid plaque deposition in the brain. The safety, tolerability, and pharmacodynamic effects of Alzhemed were assessed in a double-blind study in which 58 individuals with mild-to-moderate AD (MMSE 13-25) were randomized to receive placebo or Alzhemed 50, 100 or 150 mg BID for 3 months. At the end of the double-blind phase, 42 of these subjects entered a 36-month open-label (OL) phase in which they received Alzhemed 150 mg BID. Assessments included plasma and cerebrospinal fluid (CSF) Alzhemed concentrations, CSF levels of Abeta, as well as cognitive (Alzheimer's Disease Assessment Scale-cognitive subscale, Mini-Mental State Examination) and clinical performance (Clinical Dementia Rating scale, Sum-of-Boxes) measures. Alzhemed was safe and well tolerated, crossed the blood-brain barrier, and dose-dependently reduced CSF Abeta(42) levels after 3 months of treatment. Mild AD subjects (MMSE 19-25 at entry) displayed greater reduction of CSF Abeta(42) levels than moderate AD participants (MMSE 13-18 at entry). There was no effect of Alzhemed on the cognitive or clinical measures after 3 months of treatment. The OL follow-up suggested a stabilization of cognitive function especially in mild AD subjects over the 36-month study period. Alzhemed thus appears to be well tolerated with long-term exposure and reduces CSF Abeta(42) levels in mild-to-moderate AD subjects. These findings will be discussed in the context of two large-scale randomized, double-blind, placebo-controlled Phase III clinical trials that are currently being conducted to test the long-term safety and efficacy of Alzhemed. PMID- 17908053 TI - Pharmacogenetic aspects of therapy with cholinesterase inhibitors: the role of CYP2D6 in Alzheimer's disease pharmacogenetics. AB - Recent studies demonstrate that the therapeutic response in Alzheimer's disease (AD) is genotype-specific. More than 200 genes are potentially associated with AD pathogenesis and neurodegeneration, and approximately 1,400 genes distributed across the human genome account for 20 to 95% of variability in drug disposition and pharmacodynamics. Cytochrome P450 enzymes encoded by genes of the CYP superfamily, such as CYP1A1 (15q22-q24), CYP2A6 (19q13.2), CYP2C8 (10q24), CYP2C9 (10q24), CYP2C19 (10q24.1-q24.3), CYP2D6 (22q13.1), CYP2E1 (10q24.3-qter), and CYP3A5 (7q22.1), acting as terminal oxidases in multicomponent electron transfer chains which are called P450-containing monooxygenase systems, metabolize more than 90% of drugs. Some of the enzymatic products of the CYP gene superfamily can share substrates, inhibitors and inducers whereas others are quite specific for their substrates and interacting drugs. Some cholinesterase inhibitors (tacrine, donepezil, galantamine) are metabolized via CYP-related enzymes, especially CYP2D6, CYP3A4, and CYP1A2. The distribution of CYP2D6 genotypes in the Spanish population is the following: (a) Extensive Metabolizers (EM)(51.61%): *1/*1, 47.10%; and *1/*10, 4.52%; (b) Intermediate Metabolizers (IM)(32.26%): *1/*3, 1.95%; *1/*4, 17.42%; *1/*5, 3.87%; *1/*6, 2.58%; *1/*7, 0.75%; *10/*10, 1.30%; *4/*10, 3.23%; *6/*10, 0.65%; and *7/*10, 0.65%; (b) Poor Metabolizers (PM)(9.03%): *4/*4, 8.37%; and *5/*5, 0.65%; and (c) Ultrarapid Metabolizers (UM)(7.10%): *1xN/*1, 4.52%; *1xN/*4, 1.95%; and CYP2D6 gene duplications, 0.65%. PMs and UMs also accumulate genotypes of risk associated with APOE-, PS-, ACE-, and PRNP-related genes. Approximately, 15% of the AD population may exhibit an abnormal metabolism of cholinesterase inhibitors; about 50% of this population cluster would show an ultrarapid metabolism, requiring higher doses of cholinesterase inhibitors to reach a therapeutic threshold, whereas the other 50% of the cluster would exhibit a poor metabolism, displaying potential adverse events at low doses. In AD patients treated with a multifactorial therapy, including cholinesterase inhibitors (e.g., donepezil), the best responders are the CYP2D6-related EMs and IMs, and the worst responders are PMs and UMs. In addition, the presence of the APOE-4 allele in genetic clusters integrating CYP2D6 and APOE genotypes contributes to deteriorate the therapeutic outcome. From these data, it can be postulated that pharmacogenetic and pharmacogenomic factors are responsible for 75-85% of the therapeutic response in AD patients treated with conventional drugs. PMID- 17908054 TI - Mutational analysis of the FXNPXY motif within LDL receptor-related protein 1 (LRP1) reveals the functional importance of the tyrosine residues in cell growth regulation and signal transduction. AB - LRP1 [LDL (low-density lipoprotein) receptor-related protein 1]-null CHO cells (Chinese-hamster ovary cells) (13-5-1 cells) exhibited accelerated cell growth and severe tumour progression after they were xenografted into nude mice. Reconstitution of LRP1 expression in these cells, either with the full-length protein or with a minireceptor, reduced growth rate as well as suppressed tumour development. We tested the role of the tyrosine residue in the FXNPXY63 motif within the LRP1 cytoplasmic domain in signal transduction and cell growth inhibition by site-specific mutagenesis. The LRP1 minireceptors harbouring Tyr63 to alanine or Tyr63 to phenylalanine substitution had diametrically opposite effects on cell growth, cell morphology and tumour development in mice. The Y63F expressing cells showed suppressed cell growth and tumour development, which were associated with decreased beta-catenin and cadherin concentrations in the cells. On the other hand, the Y63A-expressing cells lacked inhibition on cell growth and tumour development, which were associated with hyperactivation of ERKs (extracellular-signal-regulated kinases), FAK (focal adhesion kinase) and cyclin D1 in the cells. The mutant Y63A minireceptor also exhibited reduced capacity in binding to the Dab2 (disabled 2) adaptor protein. In addition, the Y63A mutant showed increased caveolar localization, and cells expressing Y63A had altered caveolae architecture. However, tyrosine to alanine substitution at the other NPXY29 motif had no effect on cell growth or tumorigenesis. These results suggest that the FXNPXY63 motif of LRP1 not only governs cellular localization of the receptor but also exerts multiple functional effects on signalling pathways involved in cell growth regulation. PMID- 17908055 TI - Safety and immunogenicity profile of the concomitant administration of ZOSTAVAX and inactivated influenza vaccine in adults aged 50 and older. AB - OBJECTIVES: To evaluate the safety and immunogenicity of ZOSTAVAX administered concomitantly with inactivated influenza vaccine or sequentially in adults aged 50 and older. DESIGN: Randomized, blinded, placebo-controlled study. SETTING: Thirteen U.S. and seven European study sites. PARTICIPANTS: Three hundred eighty two concomitantly, 380 sequentially vaccinated subjects. INTERVENTION: The concomitant vaccination group received influenza vaccine and ZOSTAVAX at separate injection sites on Day 1 and placebo at Week 4. The nonconcomitant vaccination group received influenza vaccine and placebo at separate injection sites on Day 1 and ZOSTAVAX at Week 4. MEASUREMENTS: Primary safety endpoints: vaccine-related serious adverse experiences (AEs) within 28 days postvaccination (PV); and diary card-prompted local and systemic AEs. Primary immunogenicity endpoints: geometric mean titer (GMT) and geometric mean fold rise (GMFR) from baseline of varicella zoster virus (VZV) antibody (Ab) at 4 weeks PV according to glycoprotein enzyme linked immunosorbent assay (gpELISA) and GMT of influenza Ab for the three vaccine strains (2005-2006 influenza season) at 4 weeks PV according to hemagglutination inhibition assay. Secondary immunogenicity endpoint: influenza seroconversion rates (SCRs). RESULTS: No serious AEs related to ZOSTAVAX were observed during the study. VZV Ab GMTs 4 weeks PV for the concomitant and sequential groups were 554 and 597 gpELISA U/mL, respectively. The estimated VZV Ab GMT ratio was 0.9 (95% confidence interval (CI)=0.8-1.0), indicating noninferior (P<.001 for the null hypothesis of GMT ratio <0.67) responses. Estimated VZV Ab GMFR from baseline in the concomitant group was 2.1 (95% CI=2.0 2.3), indicating acceptable fold rise. Estimated GMT ratios (concomitant/sequential) for influenza strains A(H1N1), A(H3N2), and B were 0.9 (95% CI=0.8-1.1), 1.1 (95% CI=0.9-1.3), and 0.9 (95% CI=0.8-1.1), respectively, and SCRs were comparable across both groups, with more than 85% achieving titers of 1:40 or greater, meeting regulatory criteria. CONCLUSION: ZOSTAVAX and influenza vaccine given concomitantly are generally well tolerated in adults aged 50 and older. Ab responses were similar whether ZOSTAVAX and influenza vaccine were given concomitantly or sequentially. PMID- 17908056 TI - Early cognitive change in the general population: how do different definitions work? AB - OBJECTIVES: To explore the application of existing classifications of mild cognitive impairment (MCI) and associated states in a large population sample. DESIGN: Prospective cohort study, baseline phase (cross-sectional analysis). SETTING: Large-scale multicenter study in the United Kingdom. PARTICIPANTS: Thirteen thousand four individuals aged 65 and older from the Medical Research Council Cognitive Function and Aging Study. From this, a subsample of 2,640 individuals was selected and completed a more-detailed cognitive assessment. MEASUREMENTS: Information on sociodemographic status, general health, cognitive impairment (measured using the Mini-Mental State Examination), and functional ability was collected in a structured interview at baseline. The Geriatric Mental State Automated Geriatric Examination for Computer-Assisted Taxonomy and the Cambridge Cognitive Examination were used in assessment to determine cognitive status. Using a systematic literature review to collect all symptom classifications for nonnormal dementia states, these were then operationalized retrospectively. Each participant was classified according to each. RESULTS: Population prevalence estimates were variable (range 0.1-42%), reflecting differences in the focus and content of each state. Limited overlap existed between states such that many individuals were concurrently classified as normal and impaired. This highlights the heterogeneity in classification as captured using different definitions. CONCLUSION: Classification of cognitively impaired and cognitively normal individuals is dependent on the way criteria are defined and operationalized. Each classification captures a unique group of individuals, with little concordance. Given the importance of early detection of dementia and the calls for screening, and recruitment into pharmacological trials of cognitively impaired individuals, there is an urgent need for an agreed-upon standard MCI case definition to use as a criterion standard. PMID- 17908057 TI - Efficacy of periosteal stimulation therapy for the treatment of osteoarthritis associated chronic knee pain: an initial controlled clinical trial. AB - OBJECTIVES: To examine the efficacy of periosteal stimulation therapy (PST, osteopuncture) for the treatment of chronic pain associated with advanced knee osteoarthritis. DESIGN: Randomized, controlled clinical trial. SETTING: Outpatient pain clinic. PARTICIPANTS: Eighty-eight community-dwelling older adults with moderate knee pain or greater for 3 months or longer and Kellgren Lawrence (K-L) grade 2 through 4 radiographic severity (80% had K-L 4). INTERVENTION: Participants were randomized to receive PST or control PST once a week for 6 weeks. MEASUREMENTS: Pain severity and self-reported function (Western Ontario and McMasters University Osteoarthritis Index (WOMAC)) and physical performance (Short Physical Performance Battery (SPPB)) were assessed at baseline, after the last PST session (post), and 3 months later (follow-up). Pain severity was also assessed monthly using the multidimensional pain inventory short form. RESULTS: Pain was reduced significantly more in the PST group than in the control PST group at post (P=.003; mean WOMAC pain subscale baseline 9.4 vs 6.4) and 1 month later (P<.001), but by 2 months, pain levels had regressed to pre-intervention levels. The group-by-time interaction for the WOMAC function scale was significant at post (P=.04) but not at follow-up (P=.63). No significant group differences were found for the SPPB. Neither analgesic use nor global improvement differed between groups. There were four treatment dropouts. CONCLUSION: PST affords short-term modest pain reduction for older adults with advanced knee OA. Future research should test the effectiveness of booster treatments in sustaining analgesic benefits and of combining PST with therapeutic exercise in ameliorating disability risk. PMID- 17908058 TI - Effectiveness of education and individualized counseling in reducing environmental hazards in the homes of community-dwelling older women. AB - OBJECTIVES: To test the effectiveness of an education and counseling intervention on reducing environmental hazards in the homes of older women. DESIGN: Secondary analysis from a randomized, controlled trial with two arms: fall prevention program and health education program (control). Environmental hazards were assessed at baseline and immediately posttreatment (12-weeks). SETTING: Participants' homes. PARTICIPANTS: Two hundred seventy-two community-dwelling women aged 70 and older at risk for falling. INTERVENTION: The fall prevention program involved a comprehensive fall risk evaluation, exercise, education, individualized counseling, and referrals. The health education program included topics unrelated to fall prevention. With the exception of the fall risk evaluation conducted by a nurse practitioner, baccalaureate-prepared nurses carried out the interventions. MEASUREMENTS: Summed and individual scores for hazards related to the bathroom, floor surfaces, lighting, furniture, stairways, and storage areas. RESULTS: Environmental hazards were found in all homes, with a baseline mean+/-standard deviation of 10.7+/-2.6 total hazards and range of four to 17 hazards. Analysis of within-group changes indicated that the fall prevention group had significantly fewer bathroom, lighting, and total hazards after the intervention, whereas the health education group had significantly fewer bathroom hazards but more floor hazards. At follow-up, the fall prevention group had significantly fewer lighting hazards and total hazards than the health education group. CONCLUSION: Education and counseling have only modest effects in helping older women make recommended home modifications. To be most effective in reducing environmental hazards, fall prevention programs may need to provide and install safety devices. PMID- 17908059 TI - Poststroke urinary incontinence: one-year outcome and relationships with measures of attentiveness. AB - OBJECTIVES: To investigate the prognostic effect of poststroke urinary incontinence (UI) on 1-year outcome in relation to measurements of attention and mental processing speed. DESIGN: Prospective observational study. SETTING: Geriatric department (stroke and rehabilitation unit) in a university hospital. PARTICIPANTS: Two hundred thirty-five previously continent patients (median age 78) with an acute stroke. MEASUREMENTS: Clinical stroke syndromes, subtypes of UI, pre- and poststroke cognitive function and activities of daily living, computerized assessment of attention and processing speed for 110 of the participants, mortality and accommodation at 1 year. RESULTS: One hundred seventy patients remained continent, and 65 developed UI (27 with urge UI, 38 with UI with impaired awareness of the need to void (IA-UI). Patients with urge UI had poorer power of attention and speed of memory than continent patients but similar continuity of attention (P<.001, .001, and .07, respectively). Patients with IA UI performed poorer in all categories than continent and patients with urge UI (all P<.01). In regression analyses, IA-UI was the strongest predictor of mortality and nursing home residence after 1 year (odds ratio=15.7, 95% confidence interval=3.6-69.7). When deaths were excluded, IA-UI and continuity of attention remained independent risk factors. CONCLUSION: Patients with poststroke UI are less attentive than continent patients. Those with IA-UI perform poorest. Sustained attention seems important for outcome and should be taken more into account in the rehabilitation process. In patients who recognize their incontinence, attention-focused training might be the most effective measure of reestablishing bladder control. PMID- 17908061 TI - Tuberculin skin test reaction and body mass index in old age home residents in Hong Kong. AB - OBJECTIVES: To study the relationship between body mass index (BMI) and tuberculin skin test (TST) reaction in predicting the development of active tuberculosis (TB). DESIGN: A follow-up study. SETTING: Old age homes. PARTICIPANTS: Three thousand six hundred five residents who took part in a screening program for TB and had two-step TST using two units of the tuberculin PPD-RT23. MEASUREMENTS: Rate of development of active TB in these residents over an average follow-up period of 2.5+/-1.25 years. RESULTS: After one-step and two step testing, 46.3% and 69.6% of residents, respectively, had positive TST reactions (> or =10 mm). Thirty-four residents developed active TB (323 per 100,000 person-years) during follow-up. The only significant risk factors associated with development of active TB were positive TST according to one-step testing (adjusted odds ratio (OR)=2.91, 95% confidence interval (CI)=1.26-6.74) and a BMI less than 18.5 (adjusted OR=3.15, 95% CI=1.45-6.86). Residents with a BMI less than 18.5 and a negative TST also had greater risk of active TB than residents with a BMI greater than 18.5 and negative TST (adjusted OR=4.36, 95% CI=1.04-18.3), whereas those with a positive TST had the highest risk (adjusted OR=10.2, 95% CI=2.63-39.4). Two-step testing increased the sensitivity but reduced the specificity of TST in identifying active TB on follow-up. CONCLUSION: In the elderly population, interpretation of TST should take into consideration the BMI of the individual. A positive TST according to one-step but not two-step testing was useful in predicting the development of active TB on follow-up. PMID- 17908060 TI - Kidney function as a predictor of loss of lean mass in older adults: health, aging and body composition study. AB - OBJECTIVES: To assess the association between kidney function and change in body composition in older individuals. DESIGN: Prospective cohort study. SETTING: Two sites, Pittsburgh, Pennsylvania, and Memphis, Tennessee. PARTICIPANTS: Three thousand twenty-six well-functioning, participants aged 70 to 79 in the Health, Aging and Body Composition Study. MEASUREMENTS: Body composition (bone-free lean mass and fat mass) was measured using dual x-ray absorptiometry annually for 4 years. Kidney function was measured at baseline according to serum creatinine (SCr). Comorbidity and inflammatory markers were evaluated as covariates in mixed model, repeated-measures analysis. RESULTS: High SCr was associated with loss of lean mass in men but not women, with a stronger relationship in black men (P=.02 for difference between slopes for white and black men). In white men, after adjustment for age and comorbidity, higher SCr remained associated with loss of lean mass (-0.07+/-0.03 kg/y greater loss per 0.4 mg/dL (1 standard deviation (SD)), P=.009) but was attenuated after adjustment for inflammatory factors ( 0.05+/-0.03 kg/y greater loss per SD, P=.10). In black men, the relationship between SCr and loss of lean mass (-0.19+/-0.04 kg/y per SD, P<.001) persisted after adjustment for inflammation and overall weight change. CONCLUSION: Impaired kidney function may contribute to loss of lean mass in older men. Inflammation appeared to mediate the relationship in white but not black men. Future studies should strive to elucidate mechanisms linking kidney disease and muscle loss and identify treatments to minimize loss of lean mass and its functional consequences. PMID- 17908062 TI - Fear of falling in older women: a longitudinal study of incidence, persistence, and predictors. AB - OBJECTIVES: To determine longitudinal predictors of incident and persistent fear of falling (FOF) in older women. DESIGN: Longitudinal study. SETTING: Clinical research center based at a university hospital. PARTICIPANTS: One thousand two hundred eighty-two community-dwelling women aged 70 to 85. MEASUREMENTS: FOF at baseline and after 3 years of follow-up; a range of baseline demographic and clinical variables, including mobility, balance, and depression. RESULTS: FOF was present in 418 subjects (33%) at baseline, developed in 30% of women who had been free of the symptom at baseline, and was reported by a total of 46% of the sample after 3 years of follow-up. In cross-sectional multivariable analysis, baseline FOF was independently associated with a range of variables, including living alone, obesity, cognitive impairment, depression, and impairments in balance and mobility. Baseline predictors of FOF that persisted after 3 years were similar, whereas obesity and slower timed up and go test scores predicted new-onset FOF. CONCLUSION: FOF in older women is a common and persistent complaint that is caused mainly by impairments of balance and mobility. A range of social, psychological, and physical risk factors for disability are associated with persistence of FOF. These results imply that early intervention may be important for the prevention of persistent FOF. PMID- 17908063 TI - Increasing access and quality in Department of Veterans Affairs care at the end of life: a lesson in change. AB - The pursuit of a "good death" remains out of reach for many despite numerous piecemeal solutions to address the growing need for access to quality care at the end of life. In 2002, U.S. veteran deaths were at an all-time high, few Department of Veterans Affairs (VA) hospitals had inpatient palliative care services, and there was no reliable approach to meet home hospice needs. The VA embarked on a course of major change to improve veterans' care at the end of life. A coordinated plan to increase access to hospice and palliative care services was established, addressing policy development, program and staff development, collaboration with community hospices, outcomes measurement, and proving value to the organization. To determine progress and monitor resource allocation, workload and outcome measures were established in all settings. Within 3 years, the number of veterans receiving VA-paid home hospice had tripled, all VA hospitals had a palliative care team, 42% of all veterans who died as VA inpatients received a palliative care consultation, and a nationwide network of VA partnerships with community hospice agencies was established. Through a multifaceted strategic plan and a mission of honoring veterans' preferences for care at the end of life, the VA has made rapid progress in improved access to palliative care services for inpatients and outpatients. The VA's experience serves as a powerful example of the magnitude of change possible in a complex health system and a model for improving access and quality of palliative care services in other health systems. PMID- 17908064 TI - A vertically integrated geriatric curriculum improves medical student knowledge and clinical skills. AB - The objective of this study was to determine the effect of a vertically integrated curriculum intervention on the geriatric knowledge and performance in clinical skills of third-year medical students. This observational cohort study conducted at the University of Michigan Medical School evaluates the performance of 622 third-year medical students from the graduating class years of 2004 through 2007. An integrated curriculum intervention was developed and implemented for the class of 2006. Its elements included identification and tracking of geriatric learning outcomes in an individualized Web-based student portfolio, integration of geriatric content into preclinical courses, development of a geriatric functional assessment standardized patient instructor, and an experience in a geriatrics clinic during the ambulatory component of the third year internal medicine clerkship. Medical student performance was assessed on a geriatric knowledge test and during a geriatric functional assessment station administered during an Observed Structured Clinical Examination (OSCE) at the beginning of the fourth year. Student performance on the geriatric functional assessment OSCE station progressively improved from pre-intervention performance (mean performance+/-standard deviation 43+/-15% class of 2005, 62 + 15% class of 2006, 78+/-10% class of 2007; analysis of variance, P<.001). Similarly, student performance on the geriatric knowledge test was significantly better for the classes of 2006 and 2007 than for the class of 2005 (model F ratio=4.72; P<.001). In conclusion, an integrated approach to incorporating new educational geriatric objectives into the medical school curriculum leads to significant improvements in medical student knowledge and in important clinical skills in the functional assessment of older patients. PMID- 17908065 TI - The geriatrics and oncology dialogues. PMID- 17908066 TI - How do you prove quality improvement? PMID- 17908067 TI - A clarion call to rethink pressure ulcers in America. PMID- 17908068 TI - Falls in a hospital setting. PMID- 17908070 TI - A visual sign of failure of the air vent for urinary leg bags. PMID- 17908069 TI - Purple urine bag syndrome in geriatric wards: two faces of a coin? PMID- 17908071 TI - Lifestyle-related factors, alcohol consumption, and mild cognitive impairment. PMID- 17908072 TI - Sildenafil in older patients with secondary pulmonary hypertension. PMID- 17908073 TI - Acquired hemophilia in older people: a poor prognosis despite intensive care. PMID- 17908074 TI - Cri de coeur: alarming symptom in geriatrics. PMID- 17908075 TI - Contribution of gastric acid in elderly nursing home patients with cough reflex hypersensitivity. PMID- 17908076 TI - Hip fracture and depression in elderly patients: is there a sex effect? PMID- 17908077 TI - An elderly woman with primary hyperparathyroidism exhibits improvement of neurocognitive dysfunction after parathyroidectomy. PMID- 17908078 TI - Why stroke patients stop walking when talking. PMID- 17908079 TI - An unusual case of nonketotic hyperglycemia presenting as hemichorea. PMID- 17908080 TI - Circulating endothelial progenitor cell mobilization in centenarians: a powerful source for extended renal life. PMID- 17908081 TI - Association between the "puffy-hand sign" and chronic hepatitis C in a geriatric patient. PMID- 17908082 TI - Does knowledge of ethics and end-of-life issues inform choices in advance care planning scenarios? PMID- 17908084 TI - Non-STEMI or no NSTEMI? PMID- 17908085 TI - Human leucocyte antigen-B27 and ankylosing spondylitis. PMID- 17908086 TI - Usefulness of human leucocyte antigen-B27 subtypes in predicting ankylosing spondylitis: Taiwan experience. AB - BACKGROUND: Genetic factors are clearly attributed to the susceptibility of ankylosing spondylitis (AS). The human leucocyte antigen (HLA)-B27 proved to be the very useful marker for diagnosing AS. The aim of this study was to determine the prevalence of HLA-B27 subtypes in Taiwan and to investigate whether these subtypes may be of help in predicting the diagnosis of AS. METHODS: A total of 314 patients with AS and a control group of 71 subjects positive for HLA-B27 detected by flow cytometry analysis were recruited for the study. HLA-B27 subtypes were confirmed by the polymerase chain reaction-sequence-specific primers and sequence-specific oligonucleotide probing. RESULTS: Four B27 alleles were identified: B*2704, B*2705, B*2706 and B*2707. HLA-B*2704 was the predominant allele. There were significant differences in the distribution of HLA B27 subtypes between patients with AS and controls. Five of them who were homozygous for the B*2704 allele were solely found in AS group but not in controls. Statistical analysis showed that B*2704 was positively associated with AS, which suggested an increased possibility of having AS. Other HLA-B27 subtypes showed no strong correlation with AS. CONCLUSION: In the Taiwanese population, susceptibility to AS was determined by the presence of HLA-B*2704. Although B*2706 was reported to have a negative association with AS in Taiwanese, Thai and Chinese Singaporean populations, we report, in our study, two AS patients with B*2706 (0.6%). Disease heterogeneity suggests that other than genetic background, many pathogenic factors could be associated with AS. This may need to be investigated with a larger group of patients with AS and controls. PMID- 17908087 TI - One drug or two? Step-down therapy after i.v. antibiotics for community-acquired pneumonia. AB - BACKGROUND: The aim of the study was to describe the oral antibiotics prescribed as step-down therapy for patients hospitalized for community-acquired pneumonia (CAP). METHODS: A comparative audit of patient records in a Sydney teaching hospital, a district referral hospital and a regional hospital was carried out. Patients older than 15 years admitted between 1 July 2004 and 31 December 2004 with a diagnosis of CAP were identified by diagnostic code. The medical records were reviewed for patient demographics, the specialty of the attending physician, comorbidities, adverse drug events, relevant microbiological results and the antibiotic therapy prescribed for the treatment of pneumonia. Cases were randomly selected from all pneumonia admissions, with approximately equal numbers from urban and regional hospitals. One hundred and ninety-six admissions for CAP (in 193 patients) were included in this review. Patients were predominantly cared for by respiratory physicians (62%) and geriatricians (14%). Eighty-nine per cent of patients received dual antibiotic therapy on admission. RESULTS: For patients commenced on two antibiotics, 62% were prescribed two oral antibiotics after completing i.v. therapy, 27% were prescribed one oral agent and 11% were prescribed no step-down therapy. Geographic location and the presence of a documented antibiotic allergy affected prescribing practice. Neither the specialty of the attending medical officer nor the identification of a likely pathogen affected prescribing practice. CONCLUSION: Although most of the patients with CAP were initially prescribed two antibiotics, there was considerable variability in whether one, two or no oral agents were prescribed as step-down therapy. PMID- 17908088 TI - Low yield in screening patients with sporadic motor neuron disease for Kennedy disease. AB - The diagnostic yield of testing for Kennedy disease in patients diagnosed with sporadic motor neuron disease (MND) is unclear. We measured the CAG repeat lengths in the androgen receptor gene of patients with progressive limb weakness who had either upper and lower motor signs (n = 130), or lower motor neuron signs alone (n = 30). Only one patient with a long history of lower motor weakness had a repeat length in the Kennedy disease range. Testing for Kennedy disease is unlikely to benefit MND patients with upper motor neuron signs or those with a short history of lower motor signs. PMID- 17908089 TI - Two rare severe and fulminant presentations of Q fever in patients with minimal risk factors for this disease. AB - We described two rare severe and fulminant clinical presentations of acute Q fever. The first patient had severe multiorgan failure. The second patient had fever and severe cholera-like diarrhoea. Coxiella burnetii polymerase chain reaction on blood or serum can be clinically useful in the diagnosis of acute Q fever before seroconversion. PMID- 17908090 TI - Are Australasian academic physicians an endangered species? AB - It has been stated that academic medicine is in a worldwide crisis. Is this decline in hospital academic practice a predictable consequence of modern clinical practice with its emphasis on community and outpatient-based services as well as a corporate health-care ethos or does it relate to innate problems in the training process and career structure for academic clinicians? A better understanding of the barriers to involvement in academic practice, including the effect of gender, the role and effect of overseas training, expectation of further research degrees and issues pertaining to the Australian academic workplace will facilitate recruitment and retention of the next generation of academic clinicians. Physician-scientists remain highly relevant as medical practice and education evolves in the 21st century. Hospital-based academics carry out a critical role in the ongoing mentoring of trainees and junior colleagues, whose training is still largely hospital based in most specialty programmes. Academic clinicians are uniquely placed to translate the rapid advances in medical biology into the clinical sphere, by guiding and carrying out translational research as well as leading clinical studies. Academic physicians also play key leadership in relations with government and industry, in professional groups and medical colleges. Thus, there is a strong case to assess the problems facing recruitment and retention of physician-scientists in academic practice and to develop workable solutions. PMID- 17908091 TI - Unusual appearance of a rare skin lesion in a man with type 2 diabetes mellitus. PMID- 17908093 TI - Hyperammonaemic encephalopathy in plasma cell leukaemia. PMID- 17908092 TI - Unusual cause of dysphagia in an octogenarian. PMID- 17908094 TI - Acute encephalopathy as the initial symptom of CADASIL. PMID- 17908095 TI - Massive gastric bleeding: a rarely seen subacute complication of percutaneous endoscopic gastrostomy. PMID- 17908096 TI - Inability of Escherichia coli to resuscitate from the viable but nonculturable state. AB - After induction of the viable but nonculturable (VBNC) state in Escherichia coli populations, we analysed abiotic and biotic factors suggested to promote the resuscitation process. The response to the stressing conditions implied the formation of three subpopulations, culturable, VBNC and nonviable. In most adverse situations studied, the VBNC subpopulation did not represent the dominant fraction, decreasing with time. This suggests that, in most cases, the VBNC is not a successful phenotype. Combining methods of dilution and inhibition of remaining culturable cells, we designed a working protocol in order to distinguish unequivocally between regrowth and resuscitation. Reversion of abiotic factors inducing nonculturability as well as prevention of additional oxidative stress did not provoke resuscitation. Participation of biotic factors was studied by addition of supernatants from different origin without positive results. These results indicate that the E. coli strain used is not able to resuscitate from the VBNC state. VBNC cells release into the surrounding medium, and could thus aid in the survival of persisting culturable cells. The formation of a VBNC subpopulation could thus be considered as an adaptive process, designed for the benefit of the population as a whole. PMID- 17908097 TI - Factors controlling the carbon isotope fractionation of tetra- and trichloroethene during reductive dechlorination by Sulfurospirillum ssp. and Desulfitobacterium sp. strain PCE-S. AB - Carbon stable isotope fractionation of tetrachloroethene (PCE) and trichloroethene (TCE) was investigated during reductive dechlorination. Growing cells of Sulfurospirillum multivorans, Sulfurospirillum halorespirans, or Desulfitobacterium sp. strain PCE-S, the respective crude extracts and the abiotic reaction with cyanocobalamin (vitamin B(12)) were used. Fractionation of TCE (alphaC=1.0132-1.0187) by S. multivorans was more than one order of magnitude higher than values previously observed for tetrachloroethene (PCE) (alphaC=1.00042-1.0017). Similar differences in fractionation were observed during reductive dehalogenation by the close relative S. halorespirans with alphaC=1.0046-1.032 and alphaC=1.0187-1.0229 for PCE and TCE respectively. TCE carbon isotope fractionation (alphaC=1.0150) by the purified PCE-reductive dehalogenase from S. multivorans was more than one order of magnitude higher than fractionation of PCE (alphaC=1.0017). Carbon isotope fractionation of TCE by Desulfitobacterium sp. strain PCE-S (alphaC=1.0109-1.0122) as well as during the abiotic reaction with cyanocobalamin (alphaC=1.0154) was in a similar range to previously reported values for fractionation by mixed microbial cultures. In contrast with previous results with PCE, no effects due to rate limitations, uptake or transport of the substrate to the reactive site could be observed during TCE dechlorination. Our results show that prior to a mechanistic interpretation of stable isotope fractionation factors it has to be carefully verified how other factors such as uptake or transport affect the isotope fractionation during degradation experiments with microbial cultures. PMID- 17908099 TI - Overview and summary: School Health Policies and Programs Study 2006. AB - BACKGROUND: The School Health Policies and Programs Study (SHPPS) 2006 is the largest, most comprehensive assessment of school health programs in the United States ever conducted. METHODS: The Centers for Disease Control and Prevention conducts SHPPS every 6 years. In 2006, computer-assisted telephone interviews or self-administered mail questionnaires were completed by state education agency personnel in all 50 states plus the District of Columbia and among a nationally representative sample of districts (n=538). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=1103) and with a nationally representative sample of teachers of classes covering required health instruction in elementary schools and required health education courses in middle and high schools (n=912) and teachers of required physical education classes and courses (n=1194). RESULTS: SHPPS 2006 describes key school health policies and programs across all 8 school health program components: health education, physical education and activity, health services, mental health and social services, nutrition services, healthy and safe school environment, faculty and staff health promotion, and family and community involvement. SHPPS 2006 also provides data to monitor 6 Healthy People 2010 objectives. CONCLUSIONS: SHPPS 2006 is a new and important resource for school and public health practitioners, scientists, advocates, policymakers, and all those who care about the health and safety of youth and their ability to succeed academically and socially. PMID- 17908100 TI - Methods: School Health Policies and Programs Study 2006. AB - BACKGROUND: The School Health Policies and Programs Study (SHPPS) 2006 examined 8 components of school health programs: health education, physical education and activity, health services, mental health and social services, nutrition services, healthy and safe school environment, faculty and staff health promotion, and family and community involvement. All 8 components were assessed at the state, district, and school levels. Two components, health education and physical education and activity, also were assessed at the classroom level. METHODS: Computer-assisted telephone interviews or self-administered mail questionnaires were completed by state education agency personnel in all 50 states plus the District of Columbia and among a nationally representative sample of school districts (n=538). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=1103), with a nationally representative sample of teachers of required health education classes or courses (n=912), and with a nationally representative sample of teachers of required physical education classes or courses (n=1194). RESULTS: This article provides a detailed description of the development of the questionnaires; sampling; data collection; and data cleaning, weighting, and analysis. CONCLUSIONS: SHPPS 2006 is the largest and most comprehensive study of school health programs ever conducted. Fielding a study of this magnitude provides many challenges, and several recommendations for future studies emerged from the experience. PMID- 17908101 TI - Health education: results from the School Health Policies and Programs Study 2006. AB - BACKGROUND: School health education can effectively help reduce the prevalence of health-risk behaviors among students and have a positive influence on students' academic performance. This article describes the characteristics of school health education policies and programs in the United States at the state, district, school, and classroom levels. METHODS: The Centers for Disease Control and Prevention conducts the School Health Policies and Programs Study every 6 years. In 2006, computer-assisted telephone interviews or self-administered mail questionnaires were completed by state education agency personnel in all 50 states plus the District of Columbia and among a nationally representative sample of districts (n=459). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=920) and with a nationally representative sample of teachers of classes covering required health instruction in elementary schools and required health education courses in middle and high schools (n=912). RESULTS: Most states and districts had adopted a policy stating that schools will teach at least 1 of the 14 health topics, and nearly all schools required students to receive instruction on at least 1 of these topics. However, only 6.4% of elementary schools, 20.6% of middle schools, and 35.8% of high schools required instruction on all 14 topics. In support of schools, most states and districts offered staff development for those who teach health education, although the percentage of teachers of required health instruction receiving staff development was low. CONCLUSIONS: Health education has the potential to help students maintain and improve their health, prevent disease, and reduce health-related risk behaviors. However, despite signs of progress, this potential is not being fully realized, particularly at the school level. PMID- 17908102 TI - Physical education and physical activity: results from the School Health Policies and Programs Study 2006. AB - BACKGROUND: Comprehensive school-based physical activity programs consist of physical education and other physical activity opportunities including recess and other physical activity breaks, intramurals, interscholastic sports, and walk and bike to school initiatives. This article describes the characteristics of school physical education and physical activity policies and programs in the United States at the state, district, school, and classroom levels. METHODS: The Centers for Disease Control and Prevention conducts the School Health Policies and Programs Study every 6 years. In 2006, computer-assisted telephone interviews or self-administered mail questionnaires were completed by state education agency personnel in all 50 states plus the District of Columbia and among a nationally representative sample of districts (n=453). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=988) and with a nationally representative sample of teachers of required physical education classes and courses (n=1194). RESULTS: Most states and districts had adopted a policy stating that schools will teach physical education; however, few schools provided daily physical education. Additionally, many states, districts, and schools allowed students to be exempt from participating in physical education. Most schools provided some opportunities for students to be physically active outside physical education. Staff development for physical education was offered by states and districts, but physical education teachers generally did not receive staff development on a variety of important topics. CONCLUSIONS: To enhance physical education and physical activity in schools, a comprehensive approach at the state, district, school, and classroom levels is necessary. Policies, practices, and comprehensive staff development at the state and district levels might enable schools to improve opportunities for students to become physically active adults. PMID- 17908103 TI - Health services: results from the School Health Policies and Programs Study 2006. AB - BACKGROUND: The specific health services provided to students at school and the model for delivering these services vary across districts and schools. This article describes the characteristics of school health services in the United States, including state- and district-level policies and school practices. METHODS: The Centers for Disease Control and Prevention conducts the School Health Policies and Programs Study (SHPPS) every 6 years. In 2006, computer assisted telephone interviews or self-administered mail questionnaires were completed by state education agency personnel in all 50 states plus the District of Columbia and among a nationally representative sample of school districts (n=449). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=1029). RESULTS: Most US schools provided basic health services to students, but relatively few provided prevention services or more specialized health services. Although state- and district-level policies requiring school nurses or specifying maximum nurse-to-student ratios were relatively rare, 86.3% of schools had at least a part-time school nurse, and 52.4% of these schools, or 45.1% of all schools, had a nurse-to-student ratio of at least 1:750. CONCLUSIONS: SHPPS 2006 suggests that the breadth of school health services can and should be improved, but school districts need policy, legislative, and fiscal support to make this happen. Increasing the percentage of schools with sufficient school nurses is a critical step toward enabling schools to provide more services, but schools also need to enhance collaboration and linkages with community resources if schools are to be able to meet both the health and academic needs of students. PMID- 17908104 TI - Mental health and social services: results from the School Health Policies and Programs Study 2006. AB - BACKGROUND: Schools are in a unique position not only to identify mental health problems among children and adolescents but also to provide links to appropriate services. This article describes the characteristics of school mental health and social services in the United States, including state- and district-level policies and school practices. METHODS: The Centers for Disease Control and Prevention conducts the School Health Policies and Programs Study (SHPPS) every 6 years. In 2006, computer-assisted telephone interviews or self-administered mail questionnaires were completed by state education agency personnel in all 50 states and the District of Columbia and among a nationally representative sample of school districts (n=445). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=873). RESULTS: Although states and districts generally had not adopted policies stating that schools will have mental health and social services staff, 77.9% of schools had at least a part-time counselor who provided services to students. Fewer schools had school psychologists or social workers. Consequently, counseling services were more common in schools than were psychological or social services. Few schools delivered mental health and social services through school-based health centers. Arrangements with providers not located on school property were more common. CONCLUSIONS: SHPPS 2006 reveals that linkages with the community need to continue and grow to meet the mental health needs of students. Efforts must be made to build systematic state agendas for school-based mental health, emphasizing a shared responsibility among families, schools, and other community systems. PMID- 17908105 TI - Nutrition services and foods and beverages available at school: results from the School Health Policies and Programs Study 2006. AB - BACKGROUND: Schools are in a unique position to promote healthy dietary behaviors and help ensure appropriate nutrient intake. This article describes the characteristics of both school nutrition services and the foods and beverages sold outside of the school meals program in the United States, including state- and district-level policies and school practices. METHODS: The Centers for Disease Control and Prevention conducts the School Health Policies and Programs Study every 6 years. In 2006, computer-assisted telephone interviews or self administered mail questionnaires were completed by state education agency personnel in all 50 states plus the District of Columbia and among a nationally representative sample of school districts (n=445). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=944). RESULTS: Few states required schools to restrict the availability of deep-fried foods, to prohibit the sale of foods that have low nutrient density in certain venues, or to make healthful beverages available when beverages were offered. While many schools sold healthful foods and beverages outside of the school nutrition services program, many also sold items high in fat, sodium, and added sugars. CONCLUSIONS: Nutrition services program practices in many schools continue to need improvement. Districts and schools should implement more food preparation practices that reduce the total fat, saturated fat, sodium, and added sugar content of school meals. In addition, opportunities to eat and drink at school should be used to encourage greater daily consumption of fruits, vegetables, whole grains, and nonfat or low-fat dairy products. PMID- 17908106 TI - Healthy and safe school environment, Part I: Results from the School Health Policies and Programs Study 2006. AB - BACKGROUND: Policies set at the state, district, and school levels can support and enhance a healthy and safe school environment. METHODS: The Centers for Disease Control and Prevention conducts the School Health Policies and Programs Study every 6 years. In 2006, computer-assisted telephone interviews or self administered mail questionnaires were completed by state education agency personnel in all 50 states plus the District of Columbia and among a nationally representative sample of school districts (n=461). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=1025). RESULTS: Most districts had adopted a policy on the inspection and maintenance of school facilities and equipment, and most schools had inspected and provided appropriate maintenance for each type of school facility and equipment during the 12 months preceding the study. Nearly all districts and schools had a comprehensive crisis preparedness, response, and recovery plan. Nearly all districts and schools prohibited tobacco, alcohol, and illegal drug use; fighting; weapons use; and weapon possession; but when students broke rules related to those behaviors, punitive measures were taken more often than provision of supportive services. Most schools did not reschedule outdoor activities to avoid times when the sun was at peak intensity, nor did they encourage the use of sunscreen before going outside. CONCLUSIONS: To provide students with a truly healthy and safe school environment in which learning can take place, more schools need to promote a positive school climate and reduce violence, injuries, and the use of tobacco, alcohol, and other substances. States and districts need to continue to provide policy and technical assistance in support of school efforts. PMID- 17908107 TI - Healthy and safe school environment, Part II, Physical school environment: results from the School Health Policies and Programs Study 2006. AB - BACKGROUND: As society continues to focus on the importance of academic achievement, the physical environment of schools should be addressed as 1 of the critical factors that influence academic outcomes. The School Health Policies and Programs Study (SHPPS) 2006 provides, for the first time, a comprehensive look at the extent to which schools have health-promoting physical school environment policies and programs. METHODS: The Centers for Disease Control and Prevention conducts the SHPPS every 6 years. In 2006, computer-assisted telephone interviews or self-administered mail questionnaires were completed by state education agency personnel in all 50 states and the District of Columbia and among a nationally representative sample of school districts (n=424). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=992). RESULTS: One third (35.4%) of districts and 51.4% of schools had an indoor air quality management program; 35.3% of districts had a school bus engine-idling reduction program; most districts and schools had a policy or plan for how to use, label, store, dispose of, and reduce the use of hazardous materials; 24.5% of states required districts or schools to follow an integrated pest management program; and 13.4% of districts had a policy to include green design when building new school buildings or renovating existing buildings. CONCLUSIONS: SHPPS 2006 results can guide education and health agency actions in developing and implementing evidence-based tools, policies, programs, and interventions to ensure a safe and healthy physical school environment. PMID- 17908108 TI - Faculty and staff health promotion: results from the School Health Policies and Programs Study 2006. AB - BACKGROUND: US schools employ an estimated 6.7 million workers and are thus an ideal setting for employee wellness programs. This article describes the characteristics of school employee wellness programs in the United States, including state-, district-, and school-level policies and programs. METHODS: The Centers for Disease Control and Prevention conducts the School Health Policies and Programs Study every 6 years. In 2006, computer-assisted telephone interviews or self-administered mail questionnaires were completed by state education agency personnel in 49 states plus the District of Columbia and among a nationally representative sample of school districts (n=445). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=873). RESULTS: During the 2 years preceding the study, 67.3% of states provided assistance to districts or schools on how to develop or implement faculty and staff health promotion activities or services. Although nearly all schools offered at least 1 health promotion service or activity, few schools offered coordinated activities and services within a comprehensive employee wellness program. During the 12 months preceding the study, none of the health screenings were offered by more than one third of schools; only a few of the health promotion activities and services were offered by more than one third of schools; about one third of schools offered physical activity programs, employee assistance programs, and subsidies or discounts for off-site health promotion activities; and only 1 in 10 schools provided health risk appraisals for faculty and staff. CONCLUSIONS: More schools should implement comprehensive employee wellness programs to improve faculty and staff health behaviors and health status. PMID- 17908109 TI - Family and community involvement in schools: results from the School Health Policies and Programs Study 2006. AB - BACKGROUND: Family and community involvement in schools is linked strongly to improvements in the academic achievement of students, better school attendance, and improved school programs and quality. METHODS: The Centers for Disease Control and Prevention conducts the School Health Policies and Programs Study every 6 years. In 2006, computer-assisted telephone interviews or self administered mail questionnaires were completed by state education agency personnel in all 50 states plus the District of Columbia and among a nationally representative sample of school districts (n=461). Computer-assisted personal interviews were conducted with personnel in a nationally representative sample of elementary, middle, and high schools (n=1029) and with a nationally representative sample of teachers of required health education classes and courses (n=912) and required physical education classes and courses (n=1194). RESULTS: Although family and community involvement in states, districts, and schools was limited, many states, districts, and schools collaborated with community groups and agencies to promote and support school health programs. More than half of districts and schools communicated information to families on school health program components. Teachers in 55.5% of required health education classes and courses and 30.8% of required physical education classes and courses gave students homework or projects that involved family members. CONCLUSIONS: Although family and community involvement occurred at all levels, many schools are not doing some of the fundamental things schools could do to increase family involvement. Improvements in family and community involvement can support school health programs in states, districts, schools, and classrooms nationwide. PMID- 17908110 TI - European specifications for global standards in medical education. PMID- 17908111 TI - 'I'm pickin' up good regressions': the governance of generalisability analyses. AB - CONTEXT: Investigators applying generalisability theory to educational research and evaluation have sometimes done so poorly. The main difficulties have related to: inadequate or non-random sampling of effects, dealing with naturalistic data, and interpreting and presenting variance components. METHODS: This paper addresses these areas of difficulty, and articulates an informal consensus amongst medical educators from Europe, Australia and the USA, who are familiar with generalisability theory. RESULTS: We make the following recommendations. Ensure that all relevant factors are sampled, and that the sampling meets the theory's assumption that the conditions represent a random and representative sample of the factor's 'universe'. Research evaluations will require large samples of each factor if they are to generalise adequately. Where feasible, conduct 2 separate studies (pilot and evaluation, or Generalisability and Decision studies). For unbalanced data, use either urgenova, or 1 of the procedures minimum norm quadratic unbiased estimator, (minque), maximum likelihood (ml) or restricted maximum likelihood (reml) in spss or sas if the data are too complex. State which mathematical procedure was used and the degrees of freedom (d.f.) of the effect estimates. If the procedure does not report d.f., re-analyse with type III sum of squares anova (anova ss III) and report these d.f. Describe and justify the regression model used. Present the raw variance components. Describe the effects that they represent in plain, non-statistical language. If standard error of measurement (SEM) or Reliability coefficients are presented, give the equations used to calculate them. Make sure that the method of reporting reliability (precision or discrimination) is appropriate to the purpose of the assessment. This will usually demand a precision indicator such as SEM. Consider a graphical presentation to combine precision and discrimination. PMID- 17908112 TI - 'Memorable patient deaths': reactions of hospital doctors and their need for support. AB - OBJECTIVES: To investigate the reactions experienced by hospital doctors following a recent memorable patient death, defined as a patient death that had occurred in the previous few months that the doctor recollected for any particular reason, the coping strategies employed to deal with these reactions, the impact of training, and the need for support in future situations. METHODS: We carried out a descriptive survey in 2 teaching hospitals and 1 district general hospital in West Yorkshire, UK. Subjects comprised 188 hospital doctors of all grades (from pre-registration house officer to consultant) who were attending 12 educational lunchtime meetings. Main outcome measures included the associations between the intensity of emotional and physical reactions measured using a categorical rating scale, and exposure to previous training, gender, seniority and medical specialty. RESULTS: Reactions of moderate to severe intensity to a patient death were experienced by 5.0-17.5% of doctors, regardless of gender, seniority or medical specialty. Perceived need for both training and increased support from team members was significantly associated with more intense reactions. Common coping strategies included talking, spending time alone and exercise. There was no relationship between respondents' exposure to previous training and the intensity of emotional or physical responses. CONCLUSIONS: Many doctors perceive that they deal with death well. In a minority of doctors, more supportive approaches are necessary that may include both proactive and reactive measures. Examples include raising awareness of support services and establishing formal training programmes, and increasing awareness among senior clinicians of the need to support some team members after a patient's death, which may include ensuring that timely access to a counsellor is provided. PMID- 17908113 TI - The majority of bold statements expressed during grand rounds lack scientific merit. AB - CONTEXT: Frequently, during grand rounds and other medical conferences, bold statements are made regarding 'exotic medical facts'. Such exotic expert opinions are frequently voiced with great conviction and are usually subsequently assimilated by junior staff as medical fact. METHODS: The level of scientific evidence for each exotic expert opinion expressed during daily grand rounds over a 4-month period was evaluated. If, following a short discussion of the statement, any doubt as to the merits of the claim persisted, the person who made the statement was asked to perform a search in the medical literature on the subject. RESULTS: In total, 25 cases of exotic expert opinion were identified during the study period. Of these, 22 statements were made by senior staff and 3 by residents. Careful review of the literature showed only 8 of the statements were actually evidence-based. In 17 cases the available literature actually contradicted the statement (n = 13) or no literature on the subject could be located (n = 4). Although opinions were most often expressed by staff members, the reviews of their merits were more often performed by residents. CONCLUSIONS: The vast majority of exotic expert opinions expressed by senior staff members during grand rounds are not evidence-based. Thus, great care must be taken to ensure that exotic expert opinion is not accepted as factual without careful review. Furthermore, this study shows that although seniority is (as expected) associated with a higher incidence of voicing exotic expert opinion, it is negatively associated with reviewing the merits of such opinion. PMID- 17908114 TI - How can we prepare medical students for theatre-based learning? AB - CONTEXT: The quality of medical undergraduate operating theatre-based teaching is variable. Preparation prior to attending theatre may support student learning. Identifying and agreeing key skills, competences and objectives for theatre-based teaching may contribute to this process of preparation. METHODS: We carried out a cross-sectional survey of consultant surgeons and students using a forced choice questionnaire containing 16 skills and competences classified as 'essential', 'desirable' or 'not appropriate', and a choice of 6 different teaching methods, scored for perceived effectiveness on a 5-point Likert scale. Questionnaire content was based on the findings from an earlier qualitative study. RESULTS: Comparative data analyses (Mann- Whitney and Kruskal-Wallis tests) were carried out using SPSS Version 14. A total of 42 consultant surgeons and 46 students completed the questionnaire (46% and 100% response rates, respectively). Knowledge of standard theatre etiquette and protocols, ability to scrub up adequately, ability to adhere to sterile procedures, awareness of risks to self, staff and patients, and appreciation of the need for careful peri-operative monitoring were considered 'essential' by the majority. Student and consultant responses differed significantly on 5 items, with students generally considering more practical skills and competences to be essential. Differences between students on medical and surgical attachments were also identified. CONCLUSIONS: Consultant surgeons and medical students agree on many aspects of the important learning points for theatre-based teaching. Compared with their teachers, students, particularly those on attachment to surgical specialties, are more ambitious - perhaps overly so - in the level of practical skills and risk awareness they expect to gain in theatre. PMID- 17908115 TI - Money matters: students' perceptions of the costs associated with placements. AB - CONTEXT: Placements are an integral component of the medical, nursing and allied health curricula. However, apart from the relocation costs associated with placements, little research on students' understandings and experiences of the financial implications of placements has been carried out. OBJECTIVES: We report on students' financial concerns associated with placements, which emerged as a main theme in a broader study we conducted on the impact of undergraduate student placement experiences on graduate practice. METHODS: We conducted a qualitative study which included focus group discussions (n = 17), individual interviews (n = 48) and written responses (n = 2) with undergraduate students (n = 103) and graduates (n = 27) from a tertiary institution in Victoria, Australia. RESULTS: Students identified that income generation and the costs associated with transport and placement location contributed to the financial burden of placements. Students also spoke of the implications of high financial strain impacting on their accumulation of debt as well as on their health and wellbeing. DISCUSSION: Our study advances our understanding of the implications of financial hardship experienced by medical, nursing and allied health students. In our study, students, regardless of their placement location, experienced increased demands and associated stress as a result of managing placements, paid employment and limited financial resources. We recommend that further quantitative research be conducted to measure the variables identified as emerging themes in this study. PMID- 17908116 TI - Components of postgraduate competence: analyses of thirty years of longitudinal data. AB - CONTEXT: The conceptualisation and measurement of competence in patient care are critical to the design of medical education programmes and outcome assessment. OBJECTIVE: We aimed to examine the major components and correlates of postgraduate competence in patient care. METHODS: A 24-item rating form with additional questions about resident doctors' performance and future residency offers was used. Study participants comprised 4560 subjects who graduated from Jefferson Medical College between 1975 and 2004. They pursued their graduate medical education in 508 hospitals. We used a longitudinal study design in which the rating form was completed by programme directors to evaluate residents at the end of the first postgraduate year. Factor analysis was used to identify the underlying components of postgraduate ratings. Multiple regression, t-test and correlational analyses were used to study the validity of the components that emerged. RESULTS: Two major components emerged, which we labelled 'Knowledge and Clinical Capabilities' and 'Professionalism', and which addressed the science and art of medicine, respectively. Performance measures during medical school, scores on medical licensing examinations, and global assessment of Medical Knowledge, Clinical Judgement and Data-gathering Skills showed higher correlations with scores on the Knowledge and Clinical Capabilities component. Global assessments of Professional Attitudes and ratings of Empathic Behaviour showed higher correlations with scores on the Professionalism component. Offers of continued residency and evaluations of desirable qualities were associated with both components. CONCLUSIONS: Psychometric support for measuring the components of Knowledge and Clinical Capabilities, and Professionalism provides an instrument to empirically evaluate educational outcomes to medical educators who are in search of such a tool. PMID- 17908117 TI - Teaching junior doctors to manage patients who somatise: is it possible in an afternoon? AB - CONTEXT: Many patients in primary care somatise psychological distress. Training general practitioners (GPs) to manage somatisation has been shown to lead to improvements in their management of these patients. However, the training has been intensive and conducted by psychiatrists, making it impractical for widespread use. The aim of this research was to determine the effectiveness of a teaching package in improving the ability of GP registrars to manage patients who somatise, when taught by GP vocational course tutors within the constraints of a general practice vocational training scheme. METHODS: This was a before-and-after training evaluation of GP registrars' skills. A total of 22 GP registrars and 6 GP course organisers were recruited from 3 GP vocational training schemes. The GP trainees had 2 videotaped consultations with trained actors role-playing patients with somatised depression, before and 1 month after training. RESULTS: There was a significant overall improvement in the ability of GP registrars to manage patients who somatise (mean scores on a 4-point Likert scale: pre-training 1.4 [standard deviation, SD, 0.6]; post-training 2.2 [SD 0.9]; P = 0.002). General practice registrars improved their ability to use a negotiating style of consultation (skill present in 8/22 pre-training, 16/22 post-training; P = 0.02) and also demonstrated more empathy during the 'consultation' after training (mean scores on a 5-point Likert scale: pre-training 2.3 [SD 1.0]; post-training 3.0 [SD 0.8]; P = 0.03). CONCLUSIONS: Using a structured training package, it is possible for GP vocational course tutors to successfully teach GP registrars to manage patients who somatise psychological distress. Given limited resources for teaching in terms of cost and time, this training package could have important implications for training medical staff. PMID- 17908118 TI - The doctor dilemma in interprofessional education and care: how and why will physicians collaborate? AB - CONTEXT: Interprofessional educational (IPE) initiatives are seen as a means to engage health care professionals in collaborative patient-centred care. Given the hierarchical nature of many clinical settings, it is important to examine how the aims of formal IPE courses intersect with the socialisation of medical students into roles of responsibility and authority. OBJECTIVES: This article aims to provide an overview of doctor barriers to collaboration and describe aspects of medical education and socialisation that may limit doctor engagement in the goals of interprofessional education. Additionally, the paper examines the nature of team function in the health care system, reviewing different conceptual models to propose a spectrum of collaborative possibilities. Finally, specific suggestions are offered to increase the impact of interprofessional education programmes in medical education. DISCUSSION: An acknowledgement of power differentials between health care providers is necessary in the development of models for shared responsibility between professions. Conceptual models of teamwork and collaboration must articulate the desired nature of interaction between professionals with different degrees of responsibility and authority. Educational programmes in areas such as professionalism and ethics have shown limited success when formal and informal curricula significantly diverge. The socialisation of medical students into the role of a responsible doctor must be balanced with training to share responsibility appropriately. Doctor collaborative capacity may be enhanced by programmes designed to develop particular skills for which there is evidence of improved patient outcomes. PMID- 17908119 TI - 'Tutoritis' in a non-clinical tutor in problem-based learning. PMID- 17908120 TI - Redefining the role of medical professionals and medical education in India. PMID- 17908121 TI - Is primary care 'real' medicine? Some medical students appear to think not. PMID- 17908122 TI - Organizational commitment as a predictor variable in nursing turnover research: literature review. AB - AIM: This paper is a report of a literature review to (1) demonstrate the predictability of organizational commitment as a variable, (2) compare organizational commitment and job satisfaction as predictor variables and (3) determine the usefulness of organizational commitment in nursing turnover research. BACKGROUND: Organizational commitment is not routinely selected as a predictor variable in nursing studies, although the evidence suggests that it is a reliable predictor. Findings from turnover studies can help determine the previous performance of organizational commitment, and be compared to those of studies using the more conventional variable of job satisfaction. METHODS: Published research studies in English were accessed for the period 1960-2006 using the CINAHL, EBSCOHealthsource Nursing, ERIC, PROQUEST, Journals@OVID, PubMed, PsychINFO, Health and Psychosocial Instruments (HAPI) and COCHRANE library databases and Business Source Premier. The search terms included nursing turnover, organizational commitment or job satisfaction. Only studies reporting mean comparisons, R(2) or beta values related to organizational commitment and turnover or turnover antecedents were included in the review. RESULTS: There were 25 studies in the final data set, with a subset of 23 studies generated to compare the variables of organizational commitment and job satisfaction. Results indicated robust indirect predictability of organizational commitment overall, with greater predictability by organizational commitment vs job satisfaction. CONCLUSION: Organizational commitment is a useful predictor of turnover in nursing research, and effective as a variable with the most direct impact on antecedents of turnover such as intent to stay. The organizational commitment variable should be routinely employed in nursing turnover research studies. PMID- 17908123 TI - Patients referred to a pain management clinic: beliefs, expectations and priorities. AB - AIM: This paper is a report of a study to explore patients' pain-beliefs and emotions at the point of referral to a pain clinic, their expectations of the clinic and their priorities for improvement in aspects of their lives affected by pain. BACKGROUND: Chronic pain is a common experience and, although the percentage of people with pain referred to pain clinics is increasing, they often experience complex journeys through the healthcare system. Patients' beliefs about pain have been shown to influence their experience of pain and treatment outcomes, with a focus on the organic cause of pain reported. METHODS: Three focus groups were convened with 18 participants. Ten statements about pain were distributed to each participant and ranked according to their priorities. The data were collected in 2002-2003. FINDINGS: The participants' beliefs were dominated by the search for a firm diagnosis and cure. Participants held three main beliefs; that the cause of the pain must be established; that other people do not believe in the pain of a person without a firm diagnosis; and that painkillers are a way of 'fobbing you off'. Participants had little knowledge and few concrete expectations of the pain clinic. Their main priorities for improvement were 'less pain', 'some pain free times', and being able to do more 'everyday things'. CONCLUSION: Staff delivering pain management services must understand patients' beliefs and expectations and explain their own perspectives in order to provide a sound basis for working together. PMID- 17908124 TI - Ambulatory orthopaedic surgery patients' knowledge expectations and perceptions of received knowledge. AB - AIM: This paper is a report of a study to compare orthopaedic ambulatory surgery patients' knowledge expectations before admission and their perceptions of received knowledge 2 weeks after discharge. BACKGROUND: Advances in technology and population ageing are driving up the number of ambulatory orthopaedic surgical procedures. Shorter hospital stays present a major challenge for patient education. METHODS: A descriptive comparative cross-sectional study (pre- and post-test) design was adopted. The data were collected from 120 consecutive patients in 2004, using the Hospital Patient's Knowledge Expectations Scale and Hospital Patient's Received Knowledge Scale. All patients participated in a preoperative education session given by a nurse. RESULTS: Patients expected more knowledge than they actually perceived that they received on all dimensions except the bio-physiological. They perceived that they received least knowledge about experiential, ethical, social and financial dimensions of knowledge. Knowledge expectations correlated with age and professional education. Perceptions of received knowledge correlated with earlier ambulatory surgery, and both expected knowledge and perceptions of received knowledge were related to the level of basic education. CONCLUSION: Patients' knowledge expectations are greater than the knowledge they perceived that they receive, and they cannot become empowered if they lack important knowledge. Further research is needed to learn about meeting patients' knowledge expectations. PMID- 17908125 TI - Families' lived experience one year after a child was diagnosed with type 1 diabetes. AB - AIM: This paper is a report of a study to elucidate families' lived experience of diabetes one year after a child was diagnosed with type 1 diabetes. BACKGROUND: The incidence of type 1 diabetes is rapidly increasing worldwide, with a shift towards younger age groups. This illness is treated by means of an intensive management regimen that often disrupts the child's usual activities and requires disease-focused behaviours from the child and his or her family. However, research elucidating families' lived experience from the perspective of all its members is sparse. METHOD: A hermeneutic phenomenological study was carried out in 2004, based on interviews one year after diagnosis with 11 consecutively chosen Swedish-speaking family members with children aged between 9 and 14 years. FINDINGS: The families described their one year of lived experience as living an ordinary yet different life. They experienced their lives to be neither particularly difficult nor as easy as they had been before the child was diagnosed with diabetes. Related themes were 'feeling acceptance yet frustration', 'being healthy yet invisibly ill', 'feeling independent yet supervised' and 'feeling confident yet insecure'. CONCLUSION: It may be helpful if healthcare professionals make use of the knowledge and experience of families living with the illness to meet their specific needs, especially when the affected child is experiencing fluctuating blood sugar levels. Thus, health promoting collaboration should be tailor-made for every individual and proceed from each family's everyday life. PMID- 17908126 TI - Opportunities and barriers to successful learning transfer: impact of critical care skills training. AB - AIM: This paper is a report of a study to assess the impact on nursing practice of critical care skills training for ward-based nurses. BACKGROUND: Following a government review in the UK of adult critical care provision, new ways of working were advocated to ensure that critical care services depended on the needs of the patient, not their location in the hospital. A re-conceptualization beyond service provision in high dependency units and intensive care units was required in order to deliver an integrated service. This has ramifications for training requirements. METHODS: Semi-structured interviews were used to explore perceived learning and learning transfer from a range of courses. The data were collected from course attendees (n = 47) and line-managers (n = 19) across two sites between 2005 and 2006. FINDINGS: Learning was closely associated with the clinical application of new skills and knowledge. Commonly, course attendees and line-managers quoted increased knowledge and confidence, better assessment skills and improved interprofessional working. Time with competency assessors, availability of expanding roles, and supernumerary time were key factors for successful learning transfer. Barriers were financial pressures on hospitals, lack of perceived relevance of the course to staff or nursing practice, and lack of time to practice skills or work with clinical skills facilitators. CONCLUSION: Course design should be a collaborative activity between education providers and commissioners to ensure the impact of training on practice. Relevance of material, time to practise skills and new learning, and organizational, rather than merely individual, support are essential for successful training interventions. PMID- 17908127 TI - Nurses' experiences of drug administration errors. AB - AIM: This paper is a report of a study to describe the experiences of nurses who had committed serious medication errors, the meaning these experiences carry, and what kind of help and support they received after committing their error. BACKGROUND: Medication administration is an important nursing task. Work overload, combined with increased numbers and dosages of medication prescribed, puts nurses at risk of making serious errors. A drug error has the potential for disastrous consequences for patients. What is sometimes disregarded is the effect on the nurse involved. The majority of research on nurses and medication errors is framed within biomedicine, law and management. METHODS: An explorative, descriptive design was adopted and 10 in-depth interviews were conducted in 2003 with nurses who had committed a medication error. The text was analysed using a phenomenological method. FINDINGS: Serious medication errors can have a great impact on nurses, both personally and professionally. Reactions from significant others were central to the final outcome for nurses who made drug errors. They wanted to share their experiences, but this required confidence and trust. Nurses were generally willing to accept responsibility for their errors. CONCLUSION: Strategies should be developed so that errors can be managed in a constructive manner, which includes exploring underlying causes and the counselling and support needs of the nurses involved. PMID- 17908128 TI - The Work-Related Quality of Life scale for healthcare workers. AB - AIM: This paper is a report of a study to develop and test the psychometric properties of the Work-Related Quality of Life scale for healthcare workers. BACKGROUND: As problems associated with stress and job satisfaction are evident for healthcare workers and nurses, a reliable tool to assess employees' quality of working life is required. However, previous research has produced inconsistent factor structures and inadequate psychometric properties for a range of quality of working life measures. This new scale expands the concept of quality of working life by incorporating a broad six-factor structure derived from a theoretical review of the field. METHOD: We used data from a 2003 survey of 953 healthcare workers. Eighty-six per cent of the sample is female and 36% had been employed by the organization for 1-5 years. Approximately 50% of workers were employed full-time. FINDINGS: Both exploratory and confirmatory factor analysis using split-half data sets produced a good fit and a reliable 23-item, six-factor measurement model of Work-Related Quality of Life. The factors generated were labelled: Job and Career Satisfaction, General Well-Being, Home-Work Interface, Stress at Work, Control at Work and Working Conditions. CONCLUSION: The Work Related Quality of Life measure is one of the most succinct yet psychometrically valid and reliable Quality of Working Life scales in the literature. We propose that it can appropriately be used in healthcare organizations to assess quality of working life. Further research is required to refine the instrument and assess its applicability to other areas. PMID- 17908129 TI - Grounded theory research: literature reviewing and reflexivity. AB - AIM: This paper is a report of a discussion of the arguments surrounding the role of the initial literature review in grounded theory. BACKGROUND: Researchers new to grounded theory may find themselves confused about the literature review, something we ourselves experienced, pointing to the need for clarity about use of the literature in grounded theory to help guide others about to embark on similar research journeys. DISCUSSION: The arguments for and against the use of a substantial topic-related initial literature review in a grounded theory study are discussed, giving examples from our own studies. The use of theoretically sampled literature and the necessity for reflexivity are also discussed. Reflexivity is viewed as the explicit quest to limit researcher effects on the data by awareness of self, something seen as integral both to the process of data collection and the constant comparison method essential to grounded theory. CONCLUSION: A researcher who is close to the field may already be theoretically sensitized and familiar with the literature on the study topic. Use of literature or any other preknowledge should not prevent a grounded theory arising from the inductive-deductive interplay which is at the heart of this method. Reflexivity is needed to prevent prior knowledge distorting the researcher's perceptions of the data. PMID- 17908130 TI - Monitoring treatment fidelity in a randomized controlled trial of a complex intervention. AB - AIM: This paper is a report of a study to describe how treatment fidelity is being enhanced and monitored, using a model from the National Institutes of Health Behavior Change Consortium. BACKGROUND: The objective of treatment fidelity is to minimize errors in interpreting research trial outcomes, and to ascribe those outcomes directly to the intervention at hand. Treatment fidelity procedures are included in trials of complex interventions to account for inferences made from study outcomes. Monitoring treatment fidelity can help improve study design, maximize reliability of results, increase statistical power, determine whether theory-based interventions are responsible for observed changes, and inform the research dissemination process. METHODS: Treatment fidelity recommendations from the Behavior Change Consortium were applied to the SPHERE study (Secondary Prevention of Heart DiseasE in GeneRal PracticE), a randomized controlled trial of a complex intervention. Procedures to enhance and monitor intervention implementation included standardizing training sessions, observing intervention consultations, structuring patient recall systems, and using written practice and patient care plans. The research nurse plays an important role in monitoring intervention implementation. FINDINGS: Several methods of applying treatment fidelity procedures to monitoring interventions are possible. The procedure used may be determined by availability of appropriate personnel, fiscal constraints, or time limits. Complex interventions are not straightforward and necessitate a monitoring process at trial stage. CONCLUSION: The Behavior Change Consortium's model of treatment fidelity is useful for structuring a system to monitor the implementation of a complex intervention, and helps to increase the reliability and validity of evaluation findings. PMID- 17908138 TI - Topical dorsal skin immersion in seawater induces apoptosis and proliferation in hairless mice. AB - Recreational and occupational exposure to seawater (SW), have increased but the effect of SW on skin has not been elucidated. The purpose of present study was to assess the effects of SW immersion on the dorsal skin in hairless mice. Adult hairless mice were individually immersed in SW for 3 h, 6 h and 12 h; then, full thickness dorsal skin of 2 cm diameter was excised for pathological examination (light microscope), apoptosis detection (terminal deoxynucleotidyl transferase mediated 2'-deoxyuridine 5'-triphosphate nick end labeling [TUNEL]) and proliferation index evaluation (immunohistochemistry). Normal and normal saline (NS)-immersed skin were used as controls. Histological examination revealed that there were randomly distributed cell deaths, presenting cell shrinkage, condensation of nuclear chromatin and eosinophilic cytoplasm in the epidermis, and neutrophil infiltration in the dermis, after SW immersion. Moreover, TUNEL showed low levels of apoptosis in normal (9.07 +/- 0.70%) and NS-immersed skin (9.99 +/- 1.22%). There was an apparent increase in the 6-h and 12-h SW immersed groups (29.90 +/- 6.85%, P < 0.01; 45.46 +/- 6.12%, P < 0.01, respectively). Ki 67 antigen was located in the basal layer of the epidermis and hair follicles, the rates of Ki-67-positive cells were 7.90 +/- 1.45% and 7.76 +/- 1.52% in normal and NS-immersed skin, respectively, and in the 12-h SW immersed group, the rate of Ki-67-positive cells reached 23.85 +/- 4.21% (threefold, P < 0.01). In each group, the rate of apoptosis was higher than that of proliferation. We conclude that SW immersion can cause time-dependent apoptosis and proliferation in the epidermis, and the overall effect of SW immersion is injury to the epidermis. PMID- 17908139 TI - Narrow-band ultraviolet B phototherapy in patients with recalcitrant nodular prurigo. AB - Management of nodular prurigo has been less than satisfactory. Conventional therapies such as systemic antihistamines and topical steroids have not been particularly successful. The effects of narrow-band ultraviolet B (NB-UVB) phototherapy in the treatment of various inflammatory dermatoses have been proven, however, no data exist on the efficacy and the duration of remission in NB-UVB monotherapy for nodular prurigo. The aim of this study was to evaluate the effect of NB-UVB phototherapy on recalcitrant nodular prurigo. NB-UVB phototherapy was performed once a week on 10 patients with recalcitrant nodular prurigo. The initial dose was 0.4 J/cm(2), and the dose was increased by 0.1 J/cm(2) for each treatment. The treatment was performed until the eruption was almost clear. In each patient, a mean cumulative dose of 23.88 J/cm(2) was applied over a mean of 24.3 irradiations. The mean maximum daily dose of ultraviolet B was 1.2 +/- 0.4 J/cm(2). NB-UVB phototherapy notably improved the eruption of nodular prurigo in all patients. Follow up at 1 year revealed that only one patient had relapsed. The remaining nine patients continued to derive long-term benefits. NB-UVB phototherapy appears to be an effective treatment for recalcitrant nodular prurigo, offering long-term benefits in the majority of those treated. PMID- 17908140 TI - Lichenoid drug eruption of nails induced by propylthiouracil. AB - We present the case of a patient who developed deformities of the fingernails and reddish nodules on the nail beds after administration of propylthiouracil (PTU) for 6 months to treat Grave's disease. Histological examination of the lesion revealed a lichenoid tissue reaction. After withdrawal of PTU, she noticed an improvement in the eruption and the growth of the nails. No recurrence of the eruption was detected after the withdrawal of PTU. Thus, we strongly suggest that this was a rare case of PTU-induced lichenoid drug eruption of nail. PMID- 17908141 TI - Nail discoloration occurring after 8 weeks of minocycline therapy. AB - Minocycline-induced nail pigmentation is an uncommon side-effect. It usually develops after years of the therapy, and coincides with other pigmented sites. We report a 73-year-old male and a 33-year-old female developing nail discoloration after 8 weeks of therapy of 100 mg minocycline twice daily. No other pigmentation was found elsewhere on the skin, mucous membranes, teeth or sclerae of them. Our cases demonstrate that nail pigmentation can occur after short-term minocycline therapy, and propose the possibility that nail discoloration may precede other pigmentary changes. PMID- 17908142 TI - Majocchi's granuloma of the face in an immunocompetent patient. AB - Majocchi's granuloma is a condition with chronic erythematous and indurated plaques that is a result of the rupture of a dermatophyte-infected infundibulum as a result of trauma. It is frequently seen on the anterior aspect of the legs of women. Herein, we present a case of Majocchi's granuloma of face, a site rarely involved, in an immunocompetent patient. Diagnosis was confirmed by histological and mycological examination. Histological examination revealed hyphae and arthrospores in the hair follicles and in the dermis with a diffuse dermal infiltrate consisting of lymphoplasmacytic cells, and focal collections of epithelioid cells, neutrophils and mild interstitial edema. Mycological examination confirmed the presence of fungus, Trichophyton rubrum, and the diagnosis of Majocchi's granuloma of the face was made. No concrete predisposing factor was found to be associated with the occurrence of the lesions on the face. However, the history of prolonged veiling of the face by a cloth by the patient, perhaps contributing to the occurrence of lesions on face, is a point of dubious significance. PMID- 17908144 TI - Pseudoxanthoma elasticum-like papillary dermal elastolysis developed in early middle age. AB - Pseudoxanthoma elasticum-like papillary dermal elastolysis is a rare acquired elastolytic disorder characterized by papules that resemble pseudoxanthoma elasticum, and it typically affects elderly women. Histopathological examination shows atrophic epidermis and band-like loss of elastic tissue in the papillary dermis. The pathogenesis is assumed to be related to intrinsic aging because it affects elderly people and shows the loss of elastic tissue. We report a case of pseudoxanthoma elasticum-like papillary dermal elastolysis in early middle age presenting typical clinical and histopathological findings. The patient was a 41 year-old woman who had had her lesions for 10 years. We propose that younger patients, hitherto unknown, can be affected by this disorder and suggest that mechanisms other than intrinsic aging are involved in its pathogenesis. PMID- 17908143 TI - Diffuse pulmonary hemorrhage as a fatal complication of Schonlein-Henoch purpura. AB - We report a case of a 69-year-old man with Schonlein-Henoch purpura who developed diffuse pulmonary hemorrhage. He complained of dyspnea and hemoptysis while he was treated with steroids for gastroenteropathy associated with Schonlein-Henoch purpura. The drop in his hemoglobin levels and diffuse ground-glass opacity at all levels of the lung fields on chest computed tomography were a significant clue to diagnosis. Despite pulse therapy, no clinical improvement of the lung was achieved, and he died on the 36th day after the admission. Although the combination of the corticosteroid and immunosuppressive agents is normally recommended, diffuse pulmonary hemorrhage could become a fatal complication in elderly Schonlein-Henoch patients. PMID- 17908145 TI - Fusion of single-photon emission computed tomography and computed tomography images of sentinel lymph nodes in extramammary Paget's disease of the scrotum. AB - We describe a new method for the fusion of single-photon emission computed tomography (SPECT) and multidetector computed tomography (MDCT) images using an acrylic resin marker containing 99 m-Tc phytate and viewing software (Fusion Viewer Version 1.0 for Windows). This method provided the fusion of SPECT and CT images in a short time and made it easy to identify the sentinel lymph nodes in the patient with extramammary Paget's disease of the scrotum. PMID- 17908146 TI - Interdigital amelanotic spindle-cell melanoma mimicking an inflammatory process due to dermatophytosis. AB - Herein, we report a rare case of amelanotic spindle-cell melanoma on the interdigit of the left fifth toe of an 83-year-old woman. She also had tinea pedis on the same part for more than 2 years, and the part in which the tumor developed had been macerated and colonized with both Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus. Histopathologically, the biopsy specimen was indistinguishable from that of nonspecific inflammatory granulation. Two biopsies could not lead us to the correct diagnosis until the totally excised specimen was evaluated with immunohistochemical analysis including S-100 and other melanocyte markers. The patient died with multiple metastases of the tumor 18 months after her first visit. This case suggests that refractory interdigital dermatophytoses should be treated by considering the possibility of concomitant malignant neoplasms, and immunohistochemical analysis is indispensable for differential diagnosis of malignant neoplasms suggesting nonspecific granulation. PMID- 17908147 TI - Systemic sclerosis complicated by ovarian cancer. PMID- 17908148 TI - Exacerbation of psoriasis in a chronic myelogenous leukemia patient treated with imatinib. PMID- 17908149 TI - Pustular psoriasis and vitiligo in a patient with Turner syndrome. PMID- 17908150 TI - Hemophagocytic syndrome induced by diaminodiphenylsulfone. PMID- 17908151 TI - Traumatic tattoo associated with jet injector (Dermojet) use. PMID- 17908152 TI - Pemphigus vulgaris following antirabies vaccination. PMID- 17908153 TI - Ranitidine-induced drug eruption: another case of papuloerythroderma? PMID- 17908154 TI - Restoration of DWF4 expression to the leaf margin of a dwf4 mutant is sufficient to restore leaf shape but not size: the role of the margin in leaf development. AB - The role of the margin in leaf development has been debated over a number of years. To investigate the molecular basis of events in the margin, we performed an enhancer trap screen to identify genes specifically expressed in this tissue. Analysis of one of these lines revealed abnormal differentiation in the margin, accompanied by an abnormal leaf size and shape. Further analysis revealed that this phenotype was due to insertion of the trap into DWF4, which encodes a key enzyme in brassinolide biosynthesis. Transcripts for this gene accumulated in a specific and dynamic pattern in the epidermis of young leaf primordia. Targeted expression of DWF4 to a subset of these cells (the leaf margin) in a dwf4 mutant background led to both restoration of differentiation of a specific group of leaf cells (margin cells) and restoration of wild-type leaf shape (but not leaf size). Ablation of these cells led to abrogation of leaf development and the formation of small round leaves. These data support the hypothesis that events in the margin play an essential role in leaf morphogenesis, and implicate brassinolide in the margin as a key mediator in the control of leaf shape, separable from a general function of this growth factor in the control of organ size. PMID- 17908155 TI - Overexpression of Arabidopsis MAP kinase kinase 7 leads to activation of plant basal and systemic acquired resistance. AB - There is a growing body of evidence indicating that mitogen-activated protein kinase (MAPK) cascades are involved in plant defense responses. Analysis of the completed Arabidopsis thaliana genome sequence has revealed the existence of 20 MAPKs, 10 MAPKKs and 60 MAPKKKs, implying a high level of complexity in MAPK signaling pathways, and making the assignment of gene functions difficult. The MAP kinase kinase 7 (MKK7) gene of Arabidopsis has previously been shown to negatively regulate polar auxin transport. Here we provide evidence that MKK7 positively regulates plant basal and systemic acquired resistance (SAR). The activation-tagged bud1 mutant, in which the expression of MKK7 is increased, accumulates elevated levels of salicylic acid (SA), exhibits constitutive pathogenesis-related (PR) gene expression, and displays enhanced resistance to both Pseudomonas syringae pv. maculicola (Psm) ES4326 and Hyaloperonospora parasitica Noco2. Both PR gene expression and disease resistance of the bud1 plants depend on SA, and partially depend on NPR1. We demonstrate that the constitutive defense response in bud1 plants is a result of the increased expression of MKK7, and requires the kinase activity of the MKK7 protein. We found that expression of the MKK7 gene in wild-type plants is induced by pathogen infection. Reducing mRNA levels of MKK7 by antisense RNA expression not only compromises basal resistance, but also blocks the induction of SAR. Intriguingly, ectopic expression of MKK7 in local tissues induces PR gene expression and resistance to Psm ES4326 in systemic tissues, indicating that activation of MKK7 is sufficient for generating the mobile signal of SAR. PMID- 17908156 TI - Visualization of phosphatidylinositol 4,5-bisphosphate in the plasma membrane of suspension-cultured tobacco BY-2 cells and whole Arabidopsis seedlings. AB - Phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)] is an important signalling lipid in mammalian cells, where it functions as a second-messenger precursor in response to agonist-dependent activation of phospholipase C (PLC) but also operates as a signalling molecule on its own. Much of the recent knowledge about it has come from a new technique to visualize PtdIns(4,5)P(2)in vivo, by expressing a green or yellow fluorescent protein (GFP or YFP) fused to the pleckstrin homology (PH) domain of human PLCdelta1 that specifically binds PtdIns(4,5)P(2). In this way, YFP-PH(PLCdelta1) has been shown to predominantly label the plasma membrane and to transiently translocate into the cytoplasm upon PLC activation in a variety of mammalian cell systems. In plants, biochemical studies have shown that PtdIns(4,5)P(2) is present in very small quantities, but knowledge of its localization and function is still very limited. In this study, we have used YFP-PH(PLCdelta1) to try monitoring PtdIns(4,5)P(2)/PLC signalling in stably-transformed tobacco Bright Yellow-2 (BY-2) cells and Arabidopsis seedlings. In both plant systems, no detrimental effects were observed, indicating that overexpression of the biosensor did not interfere with the function of PtdIns(4,5)P(2). Confocal imaging revealed that most of the YFP PH(PLCdelta1) fluorescence was present in the cytoplasm, and not in the plasma membrane as in mammalian cells. Nonetheless, four conditions were found in which YFP-PH(PLCdelta1) was concentrated at the plasma membrane: (i) upon treatment with the PLC inhibitor U73122; (ii) in response to salt stress; (iii) as a gradient at the tip of growing root hairs; (iv) during the final stage of a BY-2 cell division. We conclude that PtdIns(4,5)P(2), as in animals, is present in the plasma membrane of plants, but that its concentration in most cells is too low to be detected by YFP-PH(PLCdelta1). Hence, the reporter remains unbound in the cytosol, making it unsuitable to monitor PLC signalling. Nonetheless, YFP PH(PLCdelta1) is a valuable plant PtdIns(4,5)P(2) reporter, for it highlights specific cells and conditions where this lipid becomes abnormally concentrated in membranes, raising the question of what it is doing there. New roles for PtdIns(4,5)P(2) in plant cell signalling are discussed. PMID- 17908157 TI - The Arabidopsis TALE homeobox gene ATH1 controls floral competency through positive regulation of FLC. AB - Floral induction is controlled by a plethora of genes acting in different pathways that either repress or promote floral transition at the shoot apical meristem (SAM). During vegetative development high levels of floral repressors maintain the Arabidopsis SAM as incompetent to respond to promoting factors. Among these repressors, FLOWERING LOCUS C (FLC) is the most prominent. The processes underlying downregulation of FLC in response to environmental and developmental signals have been elucidated in considerable detail. However, the basal induction of FLC and its upregulation by FRIGIDA (FRI) are still poorly understood. Here we report the functional characterization of the ARABIDOPSIS THALIANA HOMEOBOX 1 (ATH1) gene. A function of ATH1 in floral repression is suggested by a gradual downregulation of ATH1 in the SAM prior to floral transition. Further evidence for such a function of ATH1 is provided by the vernalization-sensitive late flowering of plants that constitutively express ATH1. Analysis of lines that differ in FRI and/or FLC allele strength show that this late flowering is caused by upregulation of FLC as a result of synergism between ATH1 overexpression and FRI. Lack of ATH1, however, results in attenuated FLC levels independently of FRI, suggesting that ATH1 acts as a general activator of FLC expression. This is further corroborated by a reduction of FLC-mediated late flowering in fca-1 and fve-1 autonomous pathway backgrounds when combined with ath1. Since other floral repressors of the FLC clade are not significantly affected by ATH1, we conclude that ATH1 controls floral competency as a specific activator of FLC expression. PMID- 17908158 TI - TAC1, a major quantitative trait locus controlling tiller angle in rice. AB - A critical step during rice (Oryza sativa) cultivation is dense planting: a wider tiller angle will increase leaf shade and decrease photosynthesis efficiency, whereas a narrower tiller angle makes for more efficient plant architecture. The molecular basis of tiller angle remains unknown. This research demonstrates that tiller angle is controlled by a major quantitative trait locus, TAC1 (Tiller Angle Control 1). TAC1 was mapped to a 35-kb region on chromosome 9 using a large F(2) population from crosses between an indica rice, IR24, which displays a relatively spread-out plant architecture, and an introgressed line, IL55, derived from japonica rice Asominori, which displays a compact plant architecture with extremely erect tillers. Genetic complementation further identified the TAC1 gene, which harbors three introns in its coding region and a fourth 1.5-kb intron in the 3'-untranslated region. A mutation in the 3'-splicing site of this 1.5-kb intron from 'AGGA' to 'GGGA' decreases the level of tac1, resulting in a compact plant architecture with a tiller angle close to zero. Further sequence verification of the mutation in the 3'-splicing site of the 1.5-kb intron revealed that the tac1 mutation 'GGGA' was present in 88 compact japonica rice accessions and TAC1 with 'AGGA' was present in 21 wild rice accessions and 43 indica rice accessions, all with the spread-out form, indicating that tac1 had been extensively utilized in densely planted rice grown in high-latitude temperate areas and at high altitudes where japonica rice varieties are widely cultivated. PMID- 17908159 TI - Life style habits such as alcohol consumption and physical activity in relation to serum apoB / apoA-I ratio amongst 64-year-old women with varying degrees of glucose tolerance. AB - OBJECTIVES: To investigate how life style factors such as alcohol consumption and physical activity relate to the serum apoB / apoA-I ratio in a cohort of middle aged women with varying degrees of glucose tolerance. DESIGN: Observational, cross-sectional cohort study. SETTING: Research laboratory at a University Hospital. SUBJECTS: A screened cohort of 64-year-old postmenopausal women with varying degrees of glucose tolerance, ranging from diabetes (n = 232), impaired (n = 212) and normal (n = 191) glucose tolerance. MAIN OUTCOME MEASURE: ApoB / apoA-I ratio in relation to alcohol consumption and physical activity as assessed by questionnaires. RESULTS: Alcohol consumption and regular physical activity at high levels were inversely associated with the serum apoB / apoA-I ratio independently of confounding factors such as obesity, lipid-lowering treatment, degree of glucose tolerance and hormone replacement therapy. Alcohol seemed related to the apoB / apoA-I ratio mainly through increasing apoA-I, whereas physical activity seemed mainly related to lowering of apoB. Alcohol consumption above a daily intake of 8.9 g, i.e. less than a glass of wine was accompanied by a decrease in apoB / apoA-I ratio. CONCLUSIONS: Amongst these 64-year-old women with varying degrees of glucose tolerance, a moderate alcohol intake and regular physical exercise leading to sweating were associated with lower apoB / apoA-I ratio and these effects seem to be additive. PMID- 17908160 TI - Clinical and biochemical implications of low thyroid hormone levels (total and free forms) in euthyroid patients with chronic kidney disease. AB - OBJECTIVES: In this study, we explore the associations of decreased thyroid hormone levels with inflammation, wasting and survival in biochemically euthyroid patients with end-stage renal disease (ESRD). DESIGN: After exclusion of 23 patients with thyroid-stimulating hormone (TSH) values outside the normal range (0.1-4.5 mIU L(-1)), 187 clinically and biochemically euthyroid incident ESRD stage 5 patients starting dialysis were followed for a median of 20 (range 1-60) months. Measurements of total and free forms of thyroid hormones, s-albumin, hs CRP, interleukin (IL)-6, vascular adhesion molecule (VCAM)-1 and insulin-like growth factor 1 (IGF-1) were performed at baseline. RESULTS: In this population, 17 out of 210 patients (8%) were defined as subclinically hypothyroid. Multivariate analysis, according to receiver operating characteristic (ROC) curves, showed that mortality was best predicted by total triiodothyronine (T3). When using the cut-off levels derived from ROC, low T3 levels were associated with increased inflammation (higher hs-CRP, IL-6 and VCAM-1) and lower concentration of both s-albumin and IGF-1. Finally, low T3 but not low free triiodothyronine was associated with worse all-cause (Likelihood ratio = 45.4; P < 0.0001) and cardiovascular mortality (Likelihood ratio = 47.8; P < 0.0001) after adjustment for confounding factors. CONCLUSION: This study showed that low T3 levels are independent predictors of all-cause and also cardiovascular disease mortality in biochemically euthyroid patients, perhaps due to an intimate association with inflammation. Based on these results, the use of T3 levels in studies assessing the relationship between thyroid dysfunction and mortality risk is recommended. PMID- 17908161 TI - C-reactive protein, interleukin-6, secretory phospholipase A2 group IIA and intercellular adhesion molecule-1 in the prediction of late outcome events after acute coronary syndromes. AB - OBJECTIVE: We investigated whether levels of C-reactive protein (CRP), interleukin-6 (IL-6), secretory phospholipase A(2) group IIA (sPLA(2)-IIA) and intercellular adhesion molecule-1 (ICAM-I) predict late outcomes in patients with acute coronary syndromes (ACS). DESIGN: Prospective longitudinal study. CRP (mg L(-1)), IL-6 (pg mL(-1)), sPLA(2)-IIA (ng mL(-1)) and ICAM-1 (ng mL(-1)) were measured at days 1 (n = 757) and 4 (n = 533) after hospital admission for ACS. Their relations to mortality and rehospitalization for myocardial infarction (MI) and congestive heart failure (CHF) were determined. SETTING: Coronary Care Unit at Sahlgrenska University Hospital, Gothenburg, Sweden. SUBJECTS: Patients with ACS alive at day 30; median follow-up 75 months. RESULTS: Survival was related to day 1 levels of all markers. After adjustment for confounders, CRP, IL-6 and ICAM 1, but not sPLA(2)-IIA, independently predicted mortality and rehospitalization for CHF. For CRP, the hazard ratio (HR) was 1.3 for mortality (95% confidence interval (CI): 1.1-1.5, P = 0.003) and 1.4 for CHF (95% CI: 1.1-1.9, P = 0.006). For IL-6, HR was 1.3 for mortality (95% CI: 1.1-1.6, P < 0.001) and 1.4 for CHF (95% CI: 1.1-1.8, P = 0.02). For ICAM-1, HR was 1.2 for mortality (95% CI: 1.0 1.4, P = 0.04) and 1.3 for CHF (95% CI: 1.0-1.7, P = 0.03). No marker predicted MI. Marker levels on day 4 provided no additional predictive value. CONCLUSIONS: In patients with ACS, CRP, IL-6, sPLA(2)-IIA and ICAM-1 are associated with long term mortality and CHF, but not reinfarction. CRP, IL-6 and ICAM-1 provide prognostic information beyond that obtained by clinical variables. PMID- 17908162 TI - Multiple safeguards against tumour development. PMID- 17908163 TI - Differential leucocyte count and the risk of future coronary artery disease in healthy men and women: the EPIC-Norfolk Prospective Population Study. AB - BACKGROUND: We examined the relationship between granulocyte, lymphocyte and monocyte counts and risk of coronary heart disease (CHD) and cardiovascular disease (CVD) in men and women. There is paucity of data on the differential leucocyte count and its relationship with the risk of CHD and CVD. METHODS: This prospective study comprised 7073 men and 9035 women who were 45-79 years of age and were residents of Norfolk. United Kingdom. RESULTS: During an average of 8 years of follow-up we identified 857 incident CHD events and 2581 CVD incident events. Increased total leucocyte count was associated with increased risk for both CHD and CVD. The highest quartile of granulocyte count was associated with increased risk when compared to lowest quartile for CHD (men HR 1.70 95% CI: 1.30 2.21; women HR 1.24 95% CI: 0.91-1.69) and for CVD (men HR 1.46 95% CI: 1.24 1.71; women HR 1.20 95% CI: 1.02-1.42). The association remained unchanged when the analyses were restricted to nonsmokers and when risk was assessed for every 1000 cells L(-1) increase in cell count. In multivariable models, despite adjusting for C-reactive protein (CRP), the granulocyte count remained an independent predictor of CHD and CVD risk, especially amongst men. Lymphocyte or monocyte counts were not significantly associated with increased risk. In all analyses, additionally adjusting for CRP did not affect the results materially. CONCLUSIONS: In conclusion, we found that the higher risk for CHD and CVD associated with increased total leucocyte count seems to be accounted for by the increased granulocyte count. PMID- 17908164 TI - Renal dysfunction predicts long-term mortality in patients with lower extremity arterial disease. AB - BACKGROUND: Patients with renal insufficiency tend to suffer from advanced atherosclerosis and exhibit a reduced life expectancy. OBJECTIVES AND DESIGN: This prospective study investigated the relation between renal dysfunction and long-term all-cause and cardiovascular mortality in a population of nonsurgical patients with lower extremity arterial disease (LEAD). SUBJECTS AND METHODS: A total of 357 patients with symptomatic LEAD underwent baseline glomerular filtration rate (GFR) estimation by the 4-variable Modification Diet in Renal Diseases equation, and were then followed for 4.2 years (range: 1-17). RESULTS: During follow-up, 131 patients died (8.6 deaths per 100 patient-years), 79 of whom (60%) from cardiovascular causes. All-cause death rates were 3.8, 6.6, and 15.5 per 100 patient-years, respectively, in the groups with normal GFR, mild reduction in GFR (60-89 mL min(-1) per 1.73 m2) and chronic kidney disease (CKD; <60 mL min(-1) per 1.73 m2; P < 0.001 by log-rank test). Compared to patients with normal renal function, the risk of all-cause and cardiovascular death was significantly higher in patients with CKD [hazard ratio, respectively, 2.23, 95% confidence interval (CI): 1.16-4.34, P = 0.017; 2.15, 95% CI: 1.05-4.43, P = 0.03]. The association of CKD with all-cause and cardiovascular mortality were independent of age, LEAD severity, cardiovascular risk factors and treatment with angiotensin-converting enzyme (ACE)-inhibitors, hypolipidaemic and antiplatelet drugs. The power of GFR in predicting all-cause death was higher than that of ankle-brachial pressure index (P = 0.029) and Framingham risk score (P < 0.0001). CONCLUSION: Chronic kidney disease strongly predicts long-term mortality in patients with symptomatic LEAD irrespective of disease severity, cardiovascular risk factors and concomitant treatments. PMID- 17908165 TI - Body mass index, alcohol, tobacco and symptomatic gallstone disease: a Swedish twin study. AB - BACKGROUND/AIMS: Both genetic and environmental factors are involved in the pathogenesis of gallstone disease (GD). We aimed to examine the association between symptomatic GD and overweight (body mass index, BMI, 25-30 kg m(-2)), obesity (BMI > 30 kg m(-2)), alcohol, smoking and smoke-free tobacco by analysing a large twin population. METHODS: The Swedish Twin Registry (STR) was linked to the Swedish Hospital Discharge and Causes of Death Registries for GD and GD surgery related diagnoses. Weight, height, use of alcohol, smoking and smoke-free tobacco were provided by STR and analysed for possible associations by conditional logistic regression. RESULTS: Overweight and obesity were associated with a significantly higher risk for symptomatic GD in the whole study population (OR 1.86 and OR 3.38; CI: 1.52-2.28 and 2.28-5.02 respectively). High alcohol consumption was associated with a lower risk for GD in the whole population (OR 0.62; CI: 0.51-0.74) with no difference between discordant monozygotic and dizygotic twins (OR 1.08 and OR 0.96; CI: 0.82-1.42 and 0.79-1.16). Smoking or smoke-free tobacco was not correlated with GD. CONCLUSION: Consistent with epidemiological studies, we found positive associations between BMI and the development of symptomatic GD. High alcohol consumption was associated with a decreased risk against GD. Tobacco use has no impact on GD. PMID- 17908166 TI - Unprecedented chromosomal diversity and behaviour modify linkage patterns and speciation potential: structural heterozygosity in an Australian spider. AB - The Huntsman spider Delena cancerides shows an extraordinary level of chromosomal diversity and meiotic complexity. Some populations form normal bivalents at male meiosis, but 14 populations form chains of chromosomes. Six of these populations form two chains, and so show segregation behaviour which is beyond our current understanding of meiotic processes. Chromosomal variation of this sort is rarely tolerated in other species, because the segregation of long chromosome chains frequently results in gametes with too many or too few chromosomes. The resulting reproductive failure may form the basis for reproductive isolation in many species, and so the mechanisms that allow D. cancerides to segregate long chromosome chains have allowed this species to maintain cohesion despite extensive chromosomal variation over its range. The effect these chromosome chains have on the population genetics of the species is discussed, and a model for the evolution of the system is proposed. PMID- 17908167 TI - Significance of chymase-dependent angiotensin II formation in the progression of human liver fibrosis. AB - AIM: Angiotensin II may contribute to liver fibrogenesis. In addition to angiotensin-converting enzyme (ACE), chymase, which is expressed by mast cells, is also known to be an angiotensin II-forming enzyme. However, it is unclear which of these two angiotensin II-forming enzymes plays a more important role in liver cirrhosis progression. In the present study, the role of angiotensin II forming enzymes in the progression of liver cirrhosis was clarified. METHODS: A total of 77 patients (16 in F0 stage, 10 in F1 stage, 22 in F2 stage, 12 in F3 stage, and 17 in F4 stage) were classified according to the new Inuyama classification into a non-cirrhosis (F0) group, an early cirrhosis (F1 + F2) group, and a chronic cirrhosis (F3 + F4) group. RESULTS: Both chymase and total angiotensin II-forming activities were significantly higher in chronic cirrhosis patients than in the other two groups. However, there was nodifference among the three groups in ACE activity. On immunohistology, the number of chymase- and angiotensin II-positive cells was significantly higher in the chronic cirrhosis group than in the non-cirrhosis and early cirrhosis groups. There were significant correlations between the number of chymase-positive cells and the number of angiotensin II-positive cells, between the number of chymase-positive cells and the degree of fibrosis, and between the number of angiotensin II positive cells and the degree of fibrosis. CONCLUSION: These results suggest that chymase-dependent angiotensin II formation may play an important role in hepatic fibrosis of patients with cirrhosis. PMID- 17908168 TI - Diagnosis of small hepatic nodules detected by surveillance ultrasound in patients with cirrhosis: Comparison between contrast-enhanced ultrasound and contrast-enhanced helical computed tomography. AB - AIM: To investigate the diagnostic value for the diagnosis of small (1-2 cm) hepatic nodules detected by surveillance ultrasound in patients with cirrhosis using contrast-enhanced ultrasound (CEUS) compared with that of contrast-enhanced helical computed tomography (CECT). METHODS: Seventy-two liver cirrhosis patients with 103 small hepatic nodules (1-2 cm) detected by surveillance ultrasound were enrolled in the present study. All patients underwent CEUS with SonoVue as well as CECT. Nodules which appeared by contrast enhancement during the arterial phase and contrast wash-out during the late phase on CEUS or CECT were diagnosed as malignant (hepatocellular carcinoma [HCC]). Histopathology obtained from biopsy or surgery served as the gold standard. RESULTS: According to the above diagnostic criteria, the sensitivity (i.e. rate of correct diagnosis of HCC) was 91.1% (51/56 HCC) for CEUS and the specificity (i.e. the rate of correct exclusion of HCC) was 87.2% (41/47 regenerative nodules [RN]).Therefore, the diagnostic accuracy of CEUS was 89.3% (92/103 all nodules). Using the same diagnostic criteria, the sensitivity, specificity and accuracy of CECT were 80.4% (45/56 HCC), 97.9% (46/47 RN), and 88.4% (91/103 all nodules). Overall, there was no significant difference between CEUS and CECT in the diagnostic confidence of small hepatic nodules. Eighty-six nodules (45 HCC and 41 RN) were correctly diagnosed by both modalities and six (five HCC and one RN) were misdiagnosed by both. CONCLUSION: The ability of CEUS in the characterization of small nodules (1 2 cm) detected by surveillance US in patients with liver cirrhosis is similar to that of CECT. PMID- 17908169 TI - Primary and secondary neural networks of auditory prepulse inhibition: a functional magnetic resonance imaging study of sensorimotor gating of the human acoustic startle response. AB - Feedforward inhibition deficits have been consistently demonstrated in a range of neuropsychiatric conditions using prepulse inhibition (PPI) of the acoustic startle eye-blink reflex when assessing sensorimotor gating. While PPI can be recorded in acutely decerebrated rats, behavioural, pharmacological and psychophysiological studies suggest the involvement of a complex neural network extending from brainstem nuclei to higher order cortical areas. The current functional magnetic resonance imaging study investigated the neural network underlying PPI and its association with electromyographically (EMG) recorded PPI of the acoustic startle eye-blink reflex in 16 healthy volunteers. A sparse imaging design was employed to model signal changes in blood oxygenation level dependent (BOLD) responses to acoustic startle probes that were preceded by a prepulse at 120 ms or 480 ms stimulus onset asynchrony or without prepulse. Sensorimotor gating was EMG confirmed for the 120-ms prepulse condition, while startle responses in the 480-ms prepulse condition did not differ from startle alone. Multiple regression analysis of BOLD contrasts identified activation in pons, thalamus, caudate nuclei, left angular gyrus and bilaterally in anterior cingulate, associated with EMG-recorded sensorimotor gating. Planned contrasts confirmed increased pons activation for startle alone vs 120-ms prepulse condition, while increased anterior superior frontal gyrus activation was confirmed for the reverse contrast. Our findings are consistent with a primary pontine circuitry of sensorimotor gating that interconnects with inferior parietal, superior temporal, frontal and prefrontal cortices via thalamus and striatum. PPI processes in the prefrontal, frontal and superior temporal cortex were functionally distinct from sensorimotor gating. PMID- 17908170 TI - Different effects of monocarboxylates on neuronal survival and beta-amyloid toxicity. AB - Glucose is a principal metabolic fuel in the central nervous system, but, when glucose is unavailable, the brain can utilize alternative metabolic substrates such as monocarboxylates to sustain brain functions. This study examined whether the replacement of glucose with monocarboxylates (particularly pyruvate and lactate) had an equivalent effect of glucose on neuronal survival in rat hippocampal organotypic slice cultures, or ameliorate the neurotoxicity induced by amyloid beta-peptide (Abeta). The possible mechanism was also explored. We found that pyruvate and lactate alone increased cell death in the hippocampal slice cultures at 24 and 48 h. Supplementation of glucose-containing culture media and Abeta-treated culture media with pyruvate, but not lactate, attenuated cell death as strong as with trolox, known as a reactive oxygen species scavenger, and niacinamide, an NAD(+) precursor, and this protective effect was reversed by alpha-cyano-4-hydroxycinnamic acid. Pyruvate significantly increased the aconitase activity and the NAD(+) levels in the hippocampal slices in the presence of Abeta, but did not maintain the ATP levels. Our results indicate that pyruvate and lactate alone cannot replace glucose as an alternative energy source to preserve the neuronal viability in the hippocampal slice cultures. Pyruvate, in the presence of glucose, improves neuronal survival in the hippocampal slice cultures and also protects neurons against Abeta-induced cell death in which mitochondrial NAD(P) redox status may play a central role. PMID- 17908171 TI - Excitotoxicity mediated by non-NMDA receptors causes distal axonopathy in long term cultured spinal motor neurons. AB - Excitotoxicity has been implicated as a potential cause of neuronal degeneration in amyotrophic lateral sclerosis (ALS). It has not been clear how excitotoxic injury leads to the hallmark pathological changes of ALS, such as the abnormal accumulation of filamentous proteins in axons. We have investigated the effects of overactivation of excitatory receptors in rodent neurons maintained in long term culture. Excitotoxicity, mediated principally via non-N-methyl-D-aspartate (NMDA) receptors, caused axonal swelling and accumulation of cytoskeletal proteins in the distal segments of the axons of cultured spinal, but not cortical, neurons. Axonopathy only occurred in spinal neurons maintained for 3 weeks in vitro, indicating that susceptibility to axonal pathology may be related to relative maturity of the neuron. Excitotoxic axonopathy was associated with the aberrant colocalization of phosphorylated and dephosphorylated neurofilament proteins, indicating that disruption to the regulation of phosphorylation of neurofilaments may lead to their abnormal accumulation. These data provide a strong link between excitotoxicity and the selective pattern of axonopathy of lower motor neurons that underlies neuronal dysfunction in ALS. PMID- 17908172 TI - Differential increase of extracellular dopamine and serotonin in the 'prefrontal cortex' and striatum of pigeons during working memory. AB - Monoamines, such as dopamine (DA) and serotonin (5-HT), play a central role in the modulation of cognitive processes at the forebrain level. Experimental and clinical studies based on dopaminergic pathology, depletion or medication indicate that DA, in particular, is involved in working memory (WM). However, it is unclear whether DA is indeed related to WM, whether its function is specific to the prefrontal cortex (PFC), and whether other modulators, such as 5-HT, might have similar functions. Therefore, the aims of this study were threefold. First, we analysed whether increased prefrontal DA release is related to WM in general or only to its short-term memory component. Second, we examined whether the DA release during cognitive tasks is specific to prefrontal areas or also occurs in the striatum. Third, we analysed whether prefrontal or striatal 5-HT release accompanies working and short-term memory. We approached these questions by using in vivo microdialysis to analyse the extracellular DA and 5-HT release in the pigeons' 'PFC' and striatum during matching-to-sample tasks with or without a delay. Here, we show that DA has no unitary function but is differentially released during working as well as short-term memory in the pigeons' 'prefrontal' cortex. Striatal DA shows an increased efflux only during WM that involves a delay component. WM is also accompanied by a 'prefrontal' but not a striatal release of 5-HT, whose efflux pattern is thus partly different to that of DA. Our findings thus show a triple dissociation between transmitters, structures and tasks within the avian 'prefronto'-striatal system. PMID- 17908173 TI - Mechanical and focal electrical stimuli applied to the skin of the index fingertip induce both inhibition and excitation in low-threshold flexor carpi radialis motor units. AB - It has been observed that mechanical stimulation of the skin of the index fingertip causes a weak short-latency inhibition followed by a strong long lasting facilitation of the flexor carpi radialis (FCR) H-reflex. Based on threshold and latency, these cutaneous reflexes are thought to be routed to motoneurons by parallel pathways. As recent studies have shown predominant inhibitory potentials in slow motoneurons and predominant excitatory potentials in faster ones, the question arises as to whether or not the two cutaneous pathways converge onto the same motoneuron. The poststimulus time histogram technique was used to investigate the changes in firing frequency of low threshold FCR motor units (MUs), induced by passive mechanical or focal electrical stimuli to the index skin. After gently tapping the finger pulp a small sharp inhibition appeared in 20 MUs. On average, inhibition started 10.2 +/ 1.6 ms from the homonymous Ia monosynaptic effect, and its central delay was estimated to be 1.2 +/- 1.6 ms. The subsequent facilitation, more consistent, had a mean latency of 13.5 +/- 1.7 ms. Inhibition and excitation were statistically significant (P < 0.05). A similar biphasic effect was observed in seven other FCR MUs, also after focal electrical stimulation of the same skin area. Comparison with the time course of the H-reflex, representing the whole population of MUs, showed striking similarities in time course and latency to the present MU effect. It is thus suggested that cutaneous spinal pathways may have a homogeneous distribution within the FCR motoneuron pool, and that the skewed distribution of cutaneous afferents onto motoneurons should be not taken as a rule. PMID- 17908175 TI - Common variants underlying cognitive ability: further evidence for association between the SNAP-25 gene and cognition using a family-based study in two independent Dutch cohorts. AB - The synaptosomal associated protein of 25 kDa (SNAP-25) gene, located on chromosome 20 p12-12p11.2 encodes a presynaptic terminal protein. SNAP-25 is differentially expressed in the brain, and primarily present in the neocortex, hippocampus, anterior thalamic nuclei, substantia nigra and cerebellar granular cells. Recently, a family-based genetic association was reported between variation in intelligence quotient (IQ) phenotypes and two intronic variants on the SNAP-25 gene. The present study is a follow-up association study in two Dutch cohorts of 371 children (mean age 12.4 years) and 391 adults (mean age 36.2 years). It examines the complete genomic region of the SNAP-25 gene to narrow down the location of causative genetic variant underlying the association. Two new variants in intron 1 (rs363043 and rs353016), close to the two previous reported variants (rs363039 and rs363050) showed association with variation in IQ phenotypes across both cohorts. All four single nucleotide polymorphisms were located in intron 1, within a region of about 13.8 kbp, and are known to affect transcription factor-binding sites. Contrary to what is expected in monogenic traits, subtle changes are postulated to influence the phenotypic outcome of complex (common) traits. As a result, functional polymorphisms in (non)coding regulatory sequences may affect spatial and temporal regulation of gene expression underlying normal cognitive variation. PMID- 17908174 TI - Involvement of astroglial ceramide in palmitic acid-induced Alzheimer-like changes in primary neurons. AB - A high-fat diet has been shown to significantly increase the risk of the development of Alzheimer's disease (AD), a neurodegenerative disease histochemically characterized by the accumulation of amyloid beta (Abeta) protein in senile plaques and hyperphosphorylated tau in neurofibrillary tangles. Previously, we have shown that saturated free fatty acids (FFAs), palmitic and stearic acids, caused increased amyloidogenesis and tau hyperphosphorylaion in primary rat cortical neurons. These FFA-induced effects observed in neurons were found to be mediated by astroglial FFA metabolism. Therefore, in the present study we investigated the basic mechanism relating astroglial FFA metabolism and AD-like changes observed in neurons. We found that palmitic acid significantly increased de-novo synthesis of ceramide in astroglia, which in turn was involved in inducing both increased production of the Abeta protein and hyperphosphorylation of the tau protein. Increased amyloidogenesis and hyperphoshorylation of tau lead to formation of the two most important pathophysiological characteristics associated with AD, Abeta or senile plaques and neurofibrillary tangles, respectively. In addition to these pathophysiological changes, AD is also characterized by certain metabolic changes; abnormal cerebral glucose metabolism is one of the distinct characteristics of AD. In this context, we found that palmitic acid significantly decreased the levels of astroglial glucose transporter (GLUT1) and down-regulated glucose uptake and lactate release by astroglia. Our present data establish an underlying mechanism by which saturated fatty acids induce AD-associated pathophysiological as well as metabolic changes, placing 'astroglial fatty acid metabolism' at the center of the pathogenic cascade in AD. PMID- 17908178 TI - Successful treatment of perifolliculitis capitis abscedens et suffodiens with combined isotretinoin and dapsone. AB - Perifolliculitis capitis abscedens et suffodiens (PCAS) is a rare scalp disease of unknown etiology which is hard to treat. It is often accompanied by scarring alopecia, acne conglobata, and recurrent fluctuant abscesses. PCAS belongs to the family of acne inversa (hidradenitis suppurativa). A 19-year-old man presented with PCAS for 2 years; multiple systemic antibiotic therapies and surgical approaches had shown no effect. Monotherapy with isotretinoin 80 mg daily for 4 weeks had not been successful. Combination therapy with dapsone 100 mg and isotretinoin 80 mg daily produced significant improvement. During 4 weeks of treatment significant clearing was achieved. Dapsone was reduced to 50 mg daily after 6 months, while isotretinoin was discontinued gradually. Now the patient is on dapsone 50 mg every other day and has remained free of recurrences for 6 months. PMID- 17908176 TI - Abnormal social behaviors in mice lacking Fgf17. AB - The fibroblast growth factor family of secreted signaling molecules is essential for patterning in the central nervous system. Fibroblast growth factor 17 (Fgf17) has been shown to contribute to regionalization of the rodent frontal cortex. To determine how Fgf17 signaling modulates behavior, both during development and in adulthood, we studied mice lacking one or two copies of the Fgf17 gene. Fgf17 deficient mice showed no abnormalities in overall physical growth, activity level, exploration, anxiety-like behaviors, motor co-ordination, motor learning, acoustic startle, prepulse inhibition, feeding, fear conditioning, aggression and olfactory exploration. However, they displayed striking deficits in several behaviors involving specific social interactions. Fgf17-deficient pups vocalized less than wild-type controls when separated from their mother and siblings. Elimination of Fgf17 also decreased the interaction of adult males with a novel ovariectomized female in a social recognition test and reduced the amount of time opposite-sex pairs spent engaged in prolonged, affiliative interactions during exploration of a novel environment. After social exploration of a novel environment, Fgf17-deficient mice showed less activation of the immediate-early gene Fos in the frontal cortex than wild-type controls. Our findings show that Fgf17 is required for several complex social behaviors and suggest that disturbances in Fgf17 signaling may contribute to neuropsychiatric diseases that affect such behaviors. PMID- 17908177 TI - Amygdala protein kinase C epsilon regulates corticotropin-releasing factor and anxiety-like behavior. AB - Corticotropin-releasing factor (CRF), its receptors, and signaling pathways that regulate CRF expression and responses are areas of intense investigation for new drugs to treat affective disorders. Here, we report that protein kinase C epsilon (PKCepsilon) null mutant mice, which show reduced anxiety-like behavior, have reduced levels of CRF messenger RNA and peptide in the amygdala. In primary amygdala neurons, a selective PKCepsilon activator, psiepsilonRACK, increased levels of pro-CRF, whereas reducing PKCepsilon levels through RNA interference blocked phorbol ester-stimulated increases in CRF. Local knockdown of amygdala PKCepsilon by RNA interference reduced anxiety-like behavior in wild-type mice. Furthermore, local infusion of CRF into the amygdala of PKCepsilon(-/-) mice increased their anxiety-like behavior. These results are consistent with a novel mechanism of PKCepsilon control over anxiety-like behavior through regulation of CRF in the amygdala. PMID- 17908179 TI - Dermoscopic features of actinic keratosis. AB - Actinic keratosis (AK) is a keratinocytic neoplasm that typically develops on sun damaged skin of elderly individuals. Only a few reports so far have described the dermoscopic diagnostic features of AK, mainly focusing on facial non-pigmented AKs. A typical feature of facial non-pigmented AK is a composite pattern named "strawberry pattern", characterized by a background erythema/red pseudonetwork consisting of unfocused, large vessels located between the hair follicles, associated with prominent follicular openings surrounded by a white halo. Dermoscopic characteristics of pigmented AK on the face include multiple slate gray to dark-brown dots and globules around the follicular ostia, annular granular pattern and brown to gray pseudonetwork. Recognizing specific dermoscopic features of AK can be useful in guiding the clinician in the differential diagnosis of AK with melanocytic skin lesions such as LM and non melanocytic lesions. Histopathologic examination should be performed whenever clinical and/or dermoscopic differential diagnosis is inconclusive. PMID- 17908181 TI - Role of a transcription factor Pax6 in the developing vertebrate olfactory system. AB - The olfactory system is responsible for capturing and processing odorant information, which significantly influences a variety of behaviors in animals. The vertebrate olfactory system consists of several neuronal components including the olfactory epithelium, olfactory bulb and olfactory cortex, which originate from distinct embryonic tissues. The transcription factor Pax6 is strongly expressed in the embryonic and postnatal olfactory systems, and regulates neuronal specification, migration and differentiation. Here we review classical and recent studies focusing on the role of Pax6 in the developing olfactory system, and highlight the cellular and molecular mechanisms underlying the highly coordinated developmental processes of the vertebrate olfactory system. PMID- 17908180 TI - A case of nodular sclerosis Hodgkin's lymphoma repeatedly relapsing in the context of composite plasma cell-hyaline vascular Castleman's disease: successful response to rituximab and radiotherapy. AB - We report the case of an Epstein-Barr virus (EBV)- and human immunodeficiency virus-serum negative patient suffering from repeatedly relapsing classical Hodgkin's Lymphoma (cHL) associated with a histological picture of plasma cell hyaline vascular (PC-HV) form of Castleman's disease (CD). The CD30- and CD15 positive, Reed-Sternberg/Hodgkin cells, only occasionally expressed the CD20 molecule, but not leukocyte common antigen and latent membrane protein-1. Single strand polymerase chain reaction failed to detect human herpesvirus 8 or EBV in the involved tissues. At the time of second relapse in July 2005, the clinical picture was characterized by a palpable right hypogastric mass, disclosed at physical exam, in the absence of other enlarged peripheral lymph nodes, subjective symptoms or laboratory profile alterations. Combined hybrid-(18)F fluorodeoxyglucose positron emission-computerized tomography (18F-FDG PET/CT) showed increased radionuclide uptake in multiple external iliac lymph nodes [standardized uptake value (SUV) of 7.4] and non-palpable left supraclavicular lymph nodes (SUV of 5.8). Relapsing cHL in the context of mixed PC-HV CD was documented in two of three surgically excised abdominal lymph nodes never previously enlarged or involved by any lymphoproliferative disease. Because of the limited disease extension and failure to induce continuous remission with previous conventional chemoradiotherapy, the patient was treated with six rituximab injections. This immunotherapy induced significant reduction in size of supraclavicular lymph nodes as evident at ultrasound (US) scan (<1 vs. 2.5 cm, post- vs. pretherapy), which was confirmed by the 18F-FDG PET/CT in October 2005, despite no modification in SUV of 4.2. 18F-FDG PET/CT also disclosed no radionuclide uptake by abdominal lymph nodes. Thus, a second course of four additional rituximab injections was given and subsequent 18F-FDG PET/CT indicated persistent, but reduced incorporation of radionuclide compared to the pretherapy value (SUV of 2.7) in the supraclavicular area and confirmed a normal metabolic activity in the iliac external lymph nodes. Because of uncertain persistent disease in the supraclavicular nodal site, involved-field radiotherapy (RT) was delivered in that area as consolidation treatment. After completion of rituximab and RT for 16 and 14 months respectively, US and 18F-FDG PET/CT exams were indicative of complete remission. This case is in concordance with previously published data suggesting that rituximab immunotherapy might be a valid option in the treatment of CD and also have a role in the management of relapsing cHL. PMID- 17908182 TI - Role of Bone morphogenetic protein 4 in zebrafish semicircular canal development. AB - Bone morphogenetic proteins (BMPs) are known to play roles in inner ear development of higher vertebrates. In zebrafish, there are several reports showing that members of the BMP family are expressed in the otic vesicle. We have isolated a novel zebrafish mutant gallery, which affects the development of the semicircular canal. Gallery merely forms the lateral and the immature anterior protrusion, and does not form posterior and ventral protrusions. We found that the expression of bmp2b and bmp4, both expressed in the normal optic vesicle at the protrusion stage, are extremely upregulated in the otic vesicle of gallery. To elucidate the role of BMPs in the development of the inner ear of zebrafish, we have applied excess BMP to the wild-type otic vesicle. The formation of protrusions was severely affected, and in some cases, they were completely lost in BMP4-treated embryos. Furthermore, the protrusions in gallery treated with Noggin were partially rescued. These data indicate that BMP4 plays an important role in the development of protrusions to form semicircular canals. PMID- 17908183 TI - The plant mannose-binding lectin NTL preserves cord blood haematopoietic stem/progenitor cells in long-term culture and enhances their ex vivo expansion. AB - Ex vivo expansion of haematopoietic stem and progenitor cells in cytokine combinations is effective in promoting differentiation and proliferation of multilineage progenitor cells, but often results in reduction of self-renewable stem cells. This study investigated the effect of a mannose-binding lectin, NTL, purified from Narcissus tazetta var. chinensis, on prolonged maintenance and expansion of cord blood CD34+ cells. Our results showed that the presence of NTL or Flt-3 ligand (FL) significantly preserved a population of early stem/progenitor cells in a serum- and cytokine-free culture for 35 d. The effect of NTL on the ex vivo expansion of CD34+ cells in the presence of stem cell factor, thrombopoietin (TPO) and FL was also investigated. NTL-enhanced expansion of early progenitors (CD34+, CD34+CD38-, mixed colony-forming units and CFU-GEMM) and committed progenitor cells (granulocyte CFU, erythroid burst-forming units/CFU and megakayocyte CFU) after 8 and 12 d of culture. Six weeks after transplanting 12 d-expanded cells to non-obese diabetic severe combined immunodeficient mice, increased engraftment of human CD45+ cells was observed in the bone marrow of animals that received NTL-treated cells. The dual functions of NTL on long-term preservation and expansion of early stem/multilineage progenitor cells could be developed for applications in various cell therapy strategies, such as the clinical expansion of CD34+ cells for transplantation. PMID- 17908184 TI - Use of the Reviscometer for measuring cosmetics-induced skin surface effects. AB - BACKGROUND/PURPOSE: The Reviscometer RVM600 that measures resonance running time (RRT) has been shown to be inversely related to the skin stiffness. However, very few publications describe the use of this instrument for testing the effect of cosmetic products. METHODS: Slight xerotic skin condition was induced by using an alkaline soap for 1 week. Skin has then been rehydrated with a lotion or further dehydrated and dried with sodium lauryl sulfate (SLS). Skin condition at the different stages of the study was evaluated by visual assessment for dryness and redness and by non-invasive methods (Corneometer, Cutometer, Reviscometer, Evaporimeter and squamometry). RESULTS: All methods showed highly significant changes after the slight drying phase with the soap usage. They illustrated skin repair after lotion treatment and further skin impairment after SLS application. Nevertheless, the Reviscometer was able to better statistically discriminate between the treatments (water, lotion, SLS) than the Cutometer . Measurement of the RRT along the transversal axis of the forearms was the most sensitive for differentiating between normal and dry skin and between the treatments on the basis of skin mechanical properties. CONCLUSION: The Reviscometer RVM600 is a sensitive instrument able to detect slight skin surface effect of cosmetics. Combined with published literature on the interpretation of RRT measurements on polymeric gel or in different skin conditions (elderly skin and photoaged skin), the Reviscometer looks to be a recommendable instrument to measure slight changes in SC stiffness/suppleness induced by cosmetic products. PMID- 17908185 TI - Laser induced autofluorescence studies of animal skin used in modeling of human cutaneous tissue spectroscopic measurements. AB - BACKGROUND/PURPOSE: Laser-induced autofluorescence spectroscopy provides excellent possibilities for medical diagnostics of different tissue pathologies including cancer. However, to create the whole picture of pathological changes, investigators collect spectral information from patients in vivo or they study different tumor models to obtain objective information for fluorescent properties of every kind of healthy and diseased tissue. Therefore, it is very important to find the most appropriate, and close to the human skin, animal samples from the fluorescence point of view, which will allow the extrapolation of the animal data to human spectroscopic diagnostics. METHODS: In the present work, we examined the autofluorescence properties of different animal skin tissues, which are considered as the most common skin models. A nitrogen laser was used as an excitation source. Samples of healthy mouse, chicken and pig skin in vivo and/or ex vivo were studied and were compared with results obtained from investigations of healthy human skin in vivo. RESULTS AND CONCLUSION: Specific features of the recorded spectra are discussed and the possible origin of the obtained fluorescence signals is proposed. Quantitative evaluation of data extrapolation for each skin type is also depicted. PMID- 17908186 TI - A long-term evaluation of erythema and pigmentation induced by ultraviolet radiations of different wavelengths. AB - BACKGROUND/AIMS: The long-term reactions of human skin by different ultraviolet (UV)-wavebands were not reported. This study was to investigate a time course of erythema and pigmentation induced by UVA-1, broadband UVA (BBUVA), narrowband UVB (NBUVB) and broadband UVB (BBUVB). METHODS: Ten volunteers participated in this study for 6 months. Four skin areas, from the back of each subject, were irradiated with two minimal erythema dose (MED) of four different UV wavelengths corresponding to UVA-1, BBUVA, NBUVB and BBUVB. RESULTS: For both UVA-1 and BBUVA, erythema and pigmentation were most pronounced immediately and 1 h after exposure. Erythema rapidly diminished but pigmentation persisted throughout the study. For both NBUVB and BBUVB, test areas reacted with erythema of maximum intensity at 1 and 2 days, respectively. A maximum tanning was reached at 3-6 days for NBUVB and 4-7 days for BBUVB, and the return toward the original point was at 1 and 3 months, respectively. CONCLUSION: Two MED of UVA produced far prolonged erythema and pigmentation than UVB. For UVA, UVA-1 and BBUVA showed similar intensity and time course of skin reaction. For UVB, erythema and pigmentation produced by NBUVB were milder in intensity and shorter in a time course than those by BBUVB. PMID- 17908187 TI - Mechanisms that play a role in the maintenance of the calcium gradient in the epidermis. AB - BACKGROUND/PURPOSE: Calcium regulates the proliferation and differentiation of keratinocytes and plays a role in restoration of the epidermal barrier function. The factors that maintain the calcium gradient in vivo are still unknown. A numerical model may give more insight into transport processes that maintain the epidermal calcium gradient. METHODS: In this study, transport of free calcium in the epidermis is described with diffusion, convection and electrophoresis. Binding and release of calcium results in equilibrium between free and bound calcium. The physiological epidermal calcium gradient as well as the calcium concentration in a damaged epidermis are modeled. RESULTS: The typical shape of the calcium gradient in the epidermis, as found in experimental studies, was maintained when separate formulations were used for free and bound calcium. Application of damage results in a decrease of the calcium concentration, especially in the upper living epidermis. Using this model, an estimate could be made about the fraction bound calcium in the epidermis. CONCLUSION: The typical shape of the gradient is predominantly determined by the bound calcium concentration. For both a normal and a damaged epidermis, the concentration of free calcium is mainly determined by electrophoresis in the living epidermis, whereas in the largest part of the stratum corneum diffusion is the most important factor. The convection that was determined by the transepidermal water loss did not have an effect on the calcium concentration. PMID- 17908188 TI - Cryoscopy: a novel enhancing method of in vivo skin imaging. AB - BACKGROUND: It is a common observation that superficial freezing of normal skin and skin tumors may create a transient superficial whitening effect. In this respect, cryoscopy refers to the direct observation by dermoscopy, with or without digital recording, of the visual alterations of the frozen tissues. AIMS: To define the optimal method of cryoscopy and to describe the cryoscopy patterns of normal skin and selected skin lesions. MATERIALS AND METHODS: The influence of (a) different cryogenic sources [solid carbon dioxide (-78.5 degrees C), liquid nitrogen (N(2), -196 degrees C), and a mixture of dimethyl ether and propane (-57 degrees C)], (b) various application methods (spraying, cotton chill tips, copper plate), and (c) freezing time was assessed with regard to clinical feasability, visualization quality, and persistance time of the whitening effect. Cryoscopy patterns of normal skin, callosities and of histologically proven seborrheic keratoses, verrucous hamartomas, molluscum contagiosum, keratoacanthomas, viral warts, condylomas, actinic keratoses, dermatofibromas, skin tags, basal cell carcinomas, angiomas, and melanocytic naevi were assessed. RESULTS: The cryoscopy images of skin highlighted the skin lines. They appeared similar regardless of the freezing source and the application method. The aspects differed according to the nature of the lesions. The cotton chill tip method provided a longer whitening period compared with the other cold sources, both in normal and lesional skin. Hence, it represented the most convenient way for performing digital recording cryoscopy. On normal skin, cryoapplication was limited to about 1.5 s due to pain, resulting in whitening times ranging from 6 to 9 s, which was too short for easy digital recording. On all studied skin tumors, a 10-s N(2) freezing time was not experienced as painful, and blanching time persisted for 20 34 s, allowing easy digital recording. The whitening time was longer with increasing freezing time on both normal and lesional skin. Every single examined normal skin site and all the skin lesions showed a strong whitening effect, except heavily cornified structures, including some keratoses, callosities, and viral warts. Increased contrast of the skin surface texture was observed in almost every studied lesion. CONCLUSION: The N(2) cotton chill tip technique appeared to be the most convenient technique for cryoscopy and provided longer whitening periods compared with the other freezing sources. Pain prevented its use on normal skin, but a series of exophytic skin lesions was conveniently accessible to cryoscopy. The differences in whitening periods of various epidermal components resulted in increased visual contrast, creating typical cryoscopy images for the different exophytic skin tumors. Cryoscopy represents a novel in vivo skin imaging technique that is rapid, non-invasive, cost-effective, and easily performed. It shows both investigative and diagnostic potentials. It is remarkable that cryoscopy pictures closely resemble those yielded by skin capacitance imaging. PMID- 17908189 TI - In vivo visualization of hyaluronic acid injection by high spatial resolution T2 parametric magnetic resonance images. AB - BACKGROUND/PURPOSE: In recent years, increasing use of injectable resorbable fillings has been reported for facial wrinkle treatment. However, the physiological processes involved such as the localization and subsequent diffusion of the injected product in skin tissues are poorly documented. This may be noninvasively achieved using quantitative magnetic resonance imaging (MRI), which is duly presented in this pilot study. METHODS: Hyaluronic acid (HA) was injected intradermally in the forearm of a young male volunteer. High-resolution MRI scans using a surface antenna were performed just after injection, and after 2, 4 and 9 months. Morphological images were compared with transverse relaxation time (T(2)) images computed from a pixel-by-pixel analysis. RESULTS: On high resolution morphological MR images the HA injection is barely visible, but with quantitative MRI the zone of injection is clearly seen. This is due to HA having a distinctly different transverse relaxation time, T(2) approximately 600 ms, compared with dermal and hypodermal tissues, 35 and 80 ms, respectively. CONCLUSION: These preliminary results demonstrate the ability of the T(2) images for in vivo visualization of the filler agent and also for characterization of tissue modifications. In addition, the diffusion and progressive degradation of the filler agent can be monitored by T(2) measurements over time. PMID- 17908190 TI - Cytokines at different stratum corneum levels in normal and sodium lauryl sulphate-irritated skin. AB - BACKGROUND/PURPOSE: Cytokines play an important role in inflammatory and repair processes occurring in the skin. The objectives of this study were to determine the amounts of cytokines and protein isolated by tape stripping in the different layers of the stratum corneum (SC), and to compare normal skin with skin exposed in vivo to the irritant sodium lauryl sulphate (SLS). METHODS: In eight volunteers, we determined the amount of total and soluble protein and also interleukin-1alpha (IL-1alpha) in pooled tape strips obtained from the upper, intermediate and lower parts of the SC. Three different types of tape were compared (Diamond , D-squame or Sentega tape). In a separate study, 20 volunteers were repeatedly exposed to 0.1% SLS over a 3-week period. The amounts of IL 1alpha, IL-1RA and IL-8 in strips obtained from the three different SC levels of SLS-exposed skin were compared with an unexposed site. RESULTS: For normal skin, the amounts of soluble protein and IL-1alpha were similar for the three tapes. Diamond tape showed the highest yield of total protein. The total protein yield per strip decreased to lower SC levels, whereas soluble protein and IL-1alpha normalized by soluble protein did not change across the SC. After SLS induced skin irritation, IL-1alpha decreased and IL-1RA and IL-8 increased at increasing depth into the SC. CONCLUSIONS: Tape stripping is a suitable method to determine SC cytokine concentrations in human skin. With this technique, it is possible to study changes in cytokine concentrations at different SC layers after skin irritation. PMID- 17908191 TI - Use of a 3-D imaging technique for non-invasive monitoring of the depth of experimentally induced wounds. AB - BACKGROUND/PURPOSE: Experimental studies of wound healing lack methods for standardized wounding and in situ depth assessment. Consequently, in this pilot study, an Erbium (Er):YAG laser has been used for wound induction with a non invasive 3-D imaging technique as an alternative to histology. METHODS: Erbium:YAG ablation of human skin ex vivo was performed with total fluences of 10, 50 and 200 J/cm(2), removing the stratum corneum, epidermis/papillary dermis and deeper dermis, respectively. Wound depth was measured with the 3-D method and histologically. RESULTS: Wound depth was proportional to fluence for both techniques : 3-D, 17.7+/-1.7, 43.9+/-16 and 245.2+/-61 microm; histology, 14.6+/ 1.7, 50.6+/-11.6 and 238+/-102 microm, respectively. CONCLUSIONS: The 3-D technique compares well with and is an improvement on histological measurement, providing true wound depth measurement, avoiding shape changes inherent with histology. Furthermore, the Er:YAG laser is a highly appropriate means of wound induction due to its rapidity and precision. PMID- 17908193 TI - Mechanism of pajama material on stratum corneum water content under mild cold conditions: explored by hierarchical linear regression. AB - OBJECTIVE: The aim of this study was to explore the effect of fabric material worn during nocturnal sleep on stratum corneum water content (SCWC) under mild cold conditions. METHOD: A controlled cross-over study has been designed to investigate systematically the effects of two kinds of pajama materials, cotton and polyester, on SCWC. Factors such as pajama fabric material, subjective perception and sleep quality were examined. Hierarchical linear regression was applied to assess the association between the factors and SCWC. RESULTS: Fabric material and subjective perception of coldness significantly predicted SCWC in the wear trial. Polyester fabric had a negative effect on SCWC compared with cotton, and subjective perception of coldness also had a negative effect on SCWC. CONCLUSION: The mechanisms by which pajama fabrics affect the SCWC could be due to the different thermal/hygroscopicity properties of the polyester and cotton fabric and the integral involvement of physiological and neurophysiological processes. PMID- 17908192 TI - Visualizing extracellular matrix and sensing fibroblasts metabolism in human dermis by nonlinear spectral imaging. AB - BACKGROUND/PURPOSE: The purpose of this work was to demonstrate the use of the nonlinear spectral imaging technique with the image-guided spectral analysis function and the extracting channels function in visualizing the extracellular maxtrix (ECM) structures and sensing fibroblasts metabolism within fresh and untreated human dermis. METHODS: A Zeiss LSM 510 META laser scanning microscopy and a femtosecond Ti: sapphire laser was used to obtain the nonlinear spectral images of human dermis. RESULTS: The nonlinear spectral imaging technique was useful in obtaining the biomorphology and biochemistry information from human dermis. By combining the image-guided spectral analysis function, intrinsic components were identified and their corresponding emission spectra can be analyzed. By integrating the extracting channels function, the information of ultrastructure of the ECM can be quantitatively obtained. CONCLUSION: The nonlinear spectral imaging technique has been demonstrated to be an effective technique for a detailed analysis and qualitative visualization of the ECM structure, and sensing of cell metabolism within fresh and untreated human dermis. Specifically, the ultrastructure of the ECM can be revealed, and collagen fibril spacing, elastic fiber diameters, the ration of collagen content over elastin can be quantitatively determined. It was found that the nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) signals excited at 830 nm provide the information for a good estimation of cellular redox ratio and a parameter (NF), the ration of NADH over FAD fluorescence signal, is used for sensing fibroblasts metabolism. With the advent of the clinical portability of typical multiphoton microscopy, the technique has the potential to be applied for in vivo studies and clinical examinations. PMID- 17908194 TI - Influence of polyol and oil concentration in cosmetic products on skin moisturization and skin surface roughness. AB - BACKGROUND/PURPOSE: The purpose of this study is to investigate the optimum combination of polyols and oils in moisturizing cosmetic products to improve the human skin moisturization and skin surface roughness. Polyols and oils are essential ingredients in skin care products, but it is still not understood how their concentrations affect their efficacy and sensory properties on human skin. We investigated the effect of polyol and oil concentration on skin properties by noninvasive methods. METHODS: The polyols consisted of glycerin and butylenes glycol in a ratio of 1:1 and the oils consisted of equal parts of hydrogenated polydecene, cethyl ethylhexanoate and pentaerythrityl tetraethylhexanoate. All cosmetic products were made in O/W emulsions in concentrations ranging from 0% to 30% for polyols and from 0% to 35% for oils. We investigated the effect on water content and skin surface roughness on the forearm after application of the cosmetic products. The skin water contents were measured by a Corneometer CM825 and the skin surface roughness by visual coring of skin surface biopsies in the scanning electron micrographs. RESULTS: In the first study, we found that the water content of the skin correlated highly with the polyol (up to 30%) and oil (up to 12%) concentrations, respectively. At two hours after application, the correlation coefficients were 0.971 and 0.985, respectively (P<0.01). Skin surface roughness not only showed a strong concentration dependence on polyols and oils (up to 6%). In the second study, we investigated the optimum combination of polyols and oils to improve the skin moisturization and skin surface roughness by the Response surface methodology. The water content of the skin surface was high in the ratio of polyol to oil (30:12 and 25:30). The skin surface roughness was improved considerably in the ratio of polyols to oil (30:6 and 30:35). CONCLUSIONS: Our results indicated that the skin surface properties were improved in the different ratios of their concentrations because they are influenced by not one ingredient but the interaction between polyols and oils. In this study, we could recommend the optimum concentration of polyols and oils to improve the skin surface properties. Further studies will be performed with other ingredients such as surfactants, lipids and so on. PMID- 17908195 TI - The barrier component and the driving force component of transepidermal water loss and their application to skin irritant tests. AB - BACKGROUND/AIM: According to Fick's law of diffusion, the rate of transepidermal water loss (TEWL) is determined by a barrier component and a driving force component. The objective of this study is to propose novel indicators for the assessment of skin irritation potential from chemicals using these components. METHODS: Before and after acetone/ether (AE), sodium lauryl sulfate (SLS) and nicotinic acid methyl ester (NME) treatments, the apparent mass transfer coefficient of water, K, of the stratum corneum (SC) and the apparent water vapor pressure, P(d), at the interface between SC and epidermis were estimated as measures of the SC barrier and the driving force, respectively. RESULTS: After AE treatment, K showed marked increase and P(d) remained approximately constant. All participants reacted to NME with erythema and the laser-Doppler flowmetry (LDF) value peaked at around 30 min. While there was a change in P(d), which parallels with LDF changes, K showed little variation. Repeated SLS treatment induced a slight increase in P(d) besides an increase in K. In the meantime, all participants reacted with subtle erythema. CONCLUSIONS: Our results suggest that not only the SC barrier but also the driving force determines variations in the TEWL rates during skin irritant tests. Together, K and P(d) will provide us with invaluable information about skin condition. PMID- 17908196 TI - Exogen hair characterization in human scalp. AB - BACKGROUND/AIM: Classically, the hair cycle is described as a sequence of three successive phases: a hair-growth phase named anagen, a regression phase or catagen and a resting phase or telogen. In rodents, it appears that the resting hair follicle population contains also a new phase that has been identified recently as the exogen phase of the hair cycle. This phase leads to the release of the telogen club and results in hair shedding. The aim of this paper is to propose a method that is applicable to humans and that is able to discriminate the two components of the resting hair population i.e. the telogen and the exogen hair follicles. METHODS: We used non-invasive approaches to entrap exogen scalp hair into silicon-based polymers. We also extracted growing and non-growing hair with a calibrated dynamometer. We characterized differences between anagen, catagen, telogen and exogen root ends with histochemical stains and with the scanning electron microscope. Furthermore, we documented all known hair-cycle stages with the contrast-enhanced phototrichogram (CE-PTG) technique. RESULTS: We demonstrated that anagen and telogen hair are firmly anchored to the hair follicle and that cohesion forces are correlated with hair thickness. On the contrary, exogen hair are passively retained within the hair follicle. Among the resting hair population, telogen clubs retain cellular elements of the outer root sheaths that are not found on exogen hair. The specificity of the new exogen collection method was documented with the simultaneous use of the CE-PTG method: indeed anagen, catagen and telogen follicles remain unaffected by the exogen extraction procedure. CONCLUSION: Exogen hair can be sampled specifically from the human scalp with a new non-invasive method. Our data suggest that the casual levels of exogen hair, in normal individuals and under the present experimental conditions, are usually less than seven hair per cm(2). PMID- 17908197 TI - Imaging of intradermal tattoos by optical coherence tomography. AB - BACKGROUND/PURPOSE: Tattoos have become increasingly popular followed by a growing demand for tattoo removal, and yet there is little knowledge and monitoring of tattoo pigment deposition in skin layers. The purpose of this pilot study is to describe optical coherence tomography image characteristics of intradermal tattoos. METHODS: We included five black tattoos in 3 female volunteers, 39, 35 and 30 years old. In vivo imaging of tattoo pigments in the skin is possible with optical coherence tomography (OCT), a novel non-invasive, in vivo optical imaging technology with a resolution and a penetration in skin high enough for visualization of tattoo pigment in the dermis. RESULTS: In optical coherence tomography images tattoo pigments clusters appear as dark, homogenous vertical columns and structures in the papillary dermis. OCT-scanned normal skin (without tattoos) appeared to be free of this dark structure. CONCLUSION: We have demonstrated that OCT can be used to visualize clusters of light absorbing pigments in a predictable manner. PMID- 17908198 TI - Strain in the nail at fingertip compression. AB - PURPOSE: Distribution of nail strain due to compressive force in the finger pulp was investigated. METHODS: Nail strain of the right index finger of 10 healthy men was measured using three strain gauges with two axes. Each nail strain was measured along the longitudinal and the transverse axis using sensor interface. Three CCD cameras recorded the compressive force and the fingertip deformation. Each subject pressed the right index finger with 30 degrees, 45 degrees and 60 degrees in the contact angle, with the compressive force increased from 0 to 14 N. RESULTS: When the contact angle was 30 degrees, the strain along the transverse axis increased according to the compressive force increased. The proximal-radial-transverse strain at compressive force of 13 N was significantly (P<0.01) larger than the proximal-ulnar-transverse strain. At the same compressive force, the distal-central-transverse strain was significantly (P<0.05) larger than the proximal-ulnar-transverse strain. The proximal-radial transverse strain was significantly (P<0.01) larger than the proximal-radial longitudinal strain. In the contact angle of 45 degrees and 60 degrees, same results were gained. CONCLUSION: Nail strain due to compressive force in the finger pulp was different between the rectangular axes, between the distal and the proximal parts of the nail, and between the radial and the ulnar sides. PMID- 17908199 TI - Unsupervised border detection in dermoscopy images. AB - BACKGROUND: As a result of the advances in skin imaging technology and the development of suitable image processing techniques, during the last decade, there has been a significant increase of interest in the computer-aided diagnosis of skin cancer. Automated border detection is one of the most important steps in this procedure as the accuracy of the subsequent steps crucially depends on the accuracy of this step. METHODS: In this article, we present an unsupervised approach to border detection in dermoscopy skin lesion images based on a modified version of the JSEG algorithm. RESULTS: The method is tested on a set of 100 dermoscopy images. The border detection error is quantified by a metric that uses manually determined borders from a dermatologist as the ground truth. The results are compared with three other automated methods and manually determined borders by a second dermatologist. CONCLUSION: The results demonstrate that the presented method achieves both fast and accurate border detection in dermoscopy images. PMID- 17908200 TI - Fluorescence--remission sensoring of skin tumours: preliminary results. AB - OBJECTIVE: Nonmelanoma skin cancer (NMSC) is one of the most common malignancies in men. Objective evaluation by digital dermoscopy, as for pigmented lesions, does not provide sufficient data to discriminate between benign and malignant lesions. Therefore, other techniques have to be developed. SETTING: Hospitalized patients of an academic teaching hospital were evaluated. PATIENTS AND METHODS: Because the simultaneous measurement of fluorescence and remission of skin is impossible, a principle of subsequent measurement of remission and fluorescence had been developed by our group. This was combined with dermoscopic imaging. VIS NIR remission spectroscopy was performed using the laboratory device TIDAS. Fluorescence spectroscopy was realized using a SKINSKAN. Fluorescence emission was detected by a highly sensitive PMT-detector. Based on this evaluation, we developed an optimized measuring device (FRIS, fluorescence-remission-imaging sensor) combining sensors for fluorescence, remission and digital imaging with a white light ring illumination, a drilled mirror and fibre optics. FRIS consists of an industrial personal computer with a touch screen combining three UV-VIS spectrometer modules and a white light source for remission measurements and referencing. Furthermore, included are a CCD coloured camera module and an LED white light ring-illumination. Fluorescence emission is realized by a UV-LED with a peak wavelength of 370 nm. System control uses Window frames and a specifically developed software Skinrem3.exe . Using this technology, we performed a pilot study in 19 patients with 30 NMSC-suspicious lesions including: actinic keratosis (n=10), basal cell carcinoma (BCC; n=16) and squamous cell carcinoma (SCC; n=4 with two in situ carcinomas). RESULTS: Reproducibility measured or FRIS by relative standard deviation of repeated spectroscopic measurements was <0.1% for remission and 2% for fluorescence. The technology was able to generate typical pattern of remission-corrected fluorescence data. The fluorescence differences at 430 nm allow a differentiation between actinic keratoses and BCC. A decrease of the corrected lesional fluorescence >2 AU indicates BCC. To substantiate the diagnostic potency of this technology, further studies are needed. CONCLUSIONS: A combination of fluorescence and remission readings of skin provides objective data in NMSC. We developed the FRIS equipment that allows a reproducible measurement and easy handling. PMID- 17908201 TI - Sub-epidermal imaging using polarized light spectroscopy for assessment of skin microcirculation. AB - BACKGROUND/AIMS: Many clinical conditions that affect the microcirculation of the skin are still diagnosed and followed up by observational methods alone in spite of the fact that non-invasive, more user-independent and objective methods are available today. Limited portability, high cost, lack of robustness and non specificity of findings are among the factors that have hampered the implementation of these methods in a clinical setting. The aim of this study is to present and evaluate a new, portable and easy-to-use imaging technology for investigation of the red blood cell (RBC) concentration in the skin microvasculature based on the method of polarization light spectroscopy using modified standard digital camera technology. METHODS: The use of orthogonal linear polarization filters over both the flash source and the detector array removes the polarization-retaining light reflected from the epidermal layer. Only the depolarized light backscattered from the papillary dermal matrix reaches the detector array. By separating the RGB color planes of an image acquired in this manner and applying a dedicated image processing algorithm, spectroscopic information about the chromophores in the dermal tissue can be attained. If the algorithm is based on a differential principle in which the normalized differences between the individual values of the red and green color plane are calculated, tissue components with similar spectral signature in both planes are suppressed, while components with different spectral signatures such as RBCs are enhanced. RESULTS: In vitro fluid models compare well with theory and computer simulations in describing a linear relationship between the imager output signal termed the tissue viability index (TiVi(index)) and RBC concentration in the physiological range of 0-4% RBC fraction of tissue volume (cc=0.997, n=20). The influence of oxygen saturation on the calculated RBC concentration is limited to within -3.9% for values within the physiological range (70-100% oxygen saturation). Monte Carlo simulations provide information about the sampling depth (about 0.5 mm on the average) of the imaging system. In vivo system evaluation based on iontophoresis of acetylcholine displays a heterogeneous pattern of vasodilatation appearing inside the electrode area after about 10 min. Topical application of methyl nicotinate and clobetasol propionate further demonstrates the capacity to document the extent and intensity of both an increase (erythema) and a decrease (blanching) in the skin RBC concentration without movement artifact and with compensation for irregularity in pigmentation. CONCLUSIONS: Polarization light spectroscopy imaging for assessment of RBC concentration in the skin microvasculature is a robust and accessible technique for the clinical setting. Additionally, the technique has pre-clinical research applications for investigation of the spatial and temporal aspects of skin erythema and blanching as well as a potential role in drug development, skin care product development and skin toxicological assessment. PMID- 17908202 TI - Growth control of Golgi phosphoinositides by reciprocal localization of sac1 lipid phosphatase and pik1 4-kinase. AB - Compartment-specific control of phosphoinositide lipids is essential for cell function. The Sac1 lipid phosphatase regulates endoplasmic reticulum (ER) and Golgi phosphatidylinositol-4-phosphate [PI(4)P] in response to nutrient levels and cell growth stages. During exponential growth, Sac1p interacts with Dpm1p at the ER but shuttles to the Golgi during starvation. Here, we report that a C terminal region in Sac1p is required for retention in the perinuclear ER, whereas the N-terminal domain is responsible for Golgi localization. We also show that starvation-induced shuttling of Sac1p to the Golgi depends on the coat protein complex II and the Rer1 adaptor protein. Starvation-induced shuttling of Sac1p to the Golgi specifically eliminates a pool of PI(4)P generated by the lipid kinase Pik1p. In addition, absence of nutrients leads to a rapid dissociation of Pik1p, together with its non-catalytical subunit Frq1p, from Golgi membranes. Reciprocal rounds of association/dissociation of the Sac1p lipid phosphatase and the Pik1p/Frq1p lipid kinase complex are responsible for growth-dependent control of Golgi phosphoinositides. Sac1p and Pik1p/Frq1p are therefore elements of a unique machinery that synchronizes ER and Golgi function in response to different growth conditions. PMID- 17908203 TI - Allelic diversity associated with aridity gradient in wild emmer wheat populations. AB - The association between allelic diversity and ecogeographical variables was studied in natural populations of wild emmer wheat [Triticum turgidum ssp. dicoccoides (Korn.) Thell.], the tetraploid progenitor of cultivated wheat. Patterns of allelic diversity in 54 microsatellite loci were analyzed in a collection of 145 wild emmer wheat accessions representing 25 populations that were sampled across naturally occurring aridity gradient in Israel and surrounding regions. The obtained results revealed that 56% of the genetic variation resided among accessions within populations, while only 44% of the variation resided between populations. An unweighted pair-group method analysis (UPGMA) tree constructed based on the microsatellite allelic diversity divided the 25 populations into six major groups. Several groups were comprised of populations that were collected in ecologically similar but geographically remote habitats. Furthermore, genetic differentiation between populations was independent of the geographical distances. An interesting evolutionary phenomenon is highlighted by the unimodal relationship between allelic diversity and annual rainfall (r = 0.74, P < 0.0002), indicating higher allelic diversity in populations originated from habitats with intermediate environmental stress (i.e. rainfall 350-550 mm year(-1)). These results show for the first time that the 'intermediate-disturbance hypothesis', explaining biological diversity at the ecosystem level, also dominates the genetic diversity within a single species, the lowest hierarchical element of the biological diversity. PMID- 17908204 TI - A unique Mycobacterium ESX-1 protein co-secretes with CFP-10/ESAT-6 and is necessary for inhibiting phagosome maturation. AB - The ESX-1 secretion system plays a critical role in the virulence of Mycobacterium tuberculosis and M. marinum. To date, three proteins are known to be secreted by ESX-1 and necessary for virulence, two of which are CFP-10 and ESAT-6. The ESX-1 secretion and the virulence mechanisms are not well understood. In this study, we have examined the M. marinum secretomes and identified four proteins specific to ESX-1. Two of those are CFP-10 and ESAT-6, and the other two are novel: MM1553 (homologous to Rv3483c) and Mh3881c (homologous to Rv3881c). We have shown that Mh3881c, CFP-10 and ESAT-6 are co-dependent for secretion. Mh3881c is being cleaved at close to the C-terminus during secretion, and the C terminal portion is critical to the co-dependent secretion, the ESAT-6 cellular levels, and interaction with ESAT-6. The co-dependent secretion is required for M. marinum intracellular growth in macrophages, where the Mh3881c C-terminal portion plays a critical role. The role of the co-dependent secretion in intracellular growth correlates with its role in inhibiting phagosome maturation. Both the secretion and the virulence defects of the Mh3881c mutant are complemented by Mh3881c or its M. tuberculosis homologue Rv3881c, suggesting that in M. tuberculosis, Rv3881c has similar functions. PMID- 17908205 TI - Glucosylceramide synthase is essential for alfalfa defensin-mediated growth inhibition but not for pathogenicity of Fusarium graminearum. AB - Antifungal defensins, MsDef1 and MtDef4, from Medicago spp., inhibit the growth of a fungal pathogen, Fusarium graminearum, at micromolar concentrations. However, molecular mechanisms by which they inhibit the growth of this fungus are not known. We have characterized a functional role of the fungal sphingolipid glucosylceramide in regulating sensitivity of the fungus to MsDef1 and MtDef4. A null mutation of the FgGCS1 gene encoding glucosylceramide synthase results in a mutant lacking glucosylceramide. The DeltaFggcs1-null mutant becomes resistant to MsDef1, but not to MtDef4. It shows a significant change in the conidial morphology and displays dramatic polar growth defect, and its mycelia are resistant to cell wall degrading enzymes. Contrary to its essential role in the pathogenicity of a human fungal pathogen, Cryptococcus neoformans, GCS1 is not required for the pathogenicity of F. graminearum. The DeltaFggcs1 mutant successfully colonizes wheat heads and corn silk, but its ability to spread in these tissues is significantly reduced as compared with the wild-type PH-1 strain. In contrast, it retains full virulence on tomato fruits and Arabidopsis thaliana floral and foliar tissues. Based on our findings, we conclude that glucosylceramide is essential for MsDef1-mediated growth inhibition of F. graminearum, but its role in fungal pathogenesis is host-dependent. PMID- 17908206 TI - YjbH is a novel negative effector of the disulphide stress regulator, Spx, in Bacillus subtilis. AB - In the soil bacterium Bacillus subtilis Spx is a key regulator that controls expression, positively or negatively, of several genes in response to certain oxidative stresses that lead to the formation of unwanted disulphide bonds. Here we characterized the yjbH gene and show that it encodes a novel effector of Spx. The yjbH gene is part of the yjbIH operon that encodes a truncated haemoglobin (YjbI) and a predicted 34 kDa cytosolic protein of unknown function (YjbH). Deletion of yjbIH or yjbH has pleiotropic effects and affects growth, sporulation and competence development. Cells lacking yjbIH display a reduced sensitivity to the thiol oxidant diamide and show an apparent down- or upregulation of several transcripts that belong to the Spx regulon. Twenty-two suppressor mutations that bypass the defects conferred by yjbH were isolated. These mutations were identified as six deletions, three nonsense and 11 missense substitutions in the spx gene. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed that mutations in yjbIH or yjbH do not affect the level of spx transcription. The combined data from the present work show that strains lacking yjbIH or yjbH overproduce Spx under unperturbed growth. The elevated Spx concentration cannot be attributed to an increased spx expression but is likely to result from control at the post-transcriptional level. YjbH is proposed to affect the cellular concentration of Spx by modulating proteolysis via the ClpXP protease. PMID- 17908207 TI - Ectopic expression of a wood-abundant expansin PttEXPA1 promotes cell expansion in primary and secondary tissues in aspen. AB - Expansins are primary agents inducing cell wall extension, and are therefore obvious targets in biotechnological applications aimed at the modification of cell size in plants. In trees, increased fibre length is a goal of both breeding and genetic engineering programmes. We used an alpha-expansin PttEXPA1 that is highly abundant in the wood-forming tissues of hybrid aspen (Populus tremula L. x P. tremuloides Michx.) to evaluate its role in fibre elongation and wood cell development. PttEXPA1 belongs to Subfamily A of alpha-expansins that have conserved motifs at the N- and C-termini of the mature protein. When PttEXPA1 was over-expressed in aspen, an extract of the cell wall-bound proteins of the transgenic plants exhibited an increased expansin activity on cellulose xyloglucan composites in vitro, indicating that PttEXPA1 is an active expansin. The transgenic lines exhibited increased stem internode elongation and leaf expansion, and larger cell sizes in the leaf epidermis, indicating that PttEXPA1 protein is capable of increasing the growth of these organs by enhancing cell wall expansion in planta. Wood cell development was also modified in the transgenic lines, but the effects were different for vessel elements and fibres, the two main cell types of aspen wood. PttEXPA1 stimulated fibre, but not vessel element, diameter growth, and marginally increased vessel element length, but did not affect fibre length. The observed differences in responsiveness to expansin of these cell types are discussed in the light of differences in their growth strategies and cell wall composition. PMID- 17908208 TI - LeuO antagonizes H-NS and StpA-dependent repression in Salmonella enterica ompS1. AB - The ompS1 gene encodes a quiescent porin in Salmonella enterica. We analysed the effects of H-NS and StpA, a paralogue of H-NS, on ompS1 expression. In an hns single mutant expression was derepressed but did not reach the maximum level. Expression in an stpA single mutant showed the same low repressed level as the wild type. In contrast, in an hns stpA background, OmpS1 became abundant in the outer membrane. The expression of ompS1 was positively regulated by LeuO, a LysR type quiescent regulator that has been involved in pathogenesis. Upon induction of the cloned leuO gene into the wild type, ompS1 was completely derepressed and the OmpS1 porin was detected in the outer membrane. LeuO activated the P1 promoter in an OmpR-dependent manner and P2 in the absence of OmpR. LeuO bound upstream of the regulatory region of ompS1 overlapping with one nucleation site of H-NS and StpA. Our results are thus consistent with a model where H-NS binds at a nucleation site and LeuO displaces H-NS and StpA. PMID- 17908210 TI - Population history of Berthelot's pipit: colonization, gene flow and morphological divergence in Macaronesia. AB - The fauna of oceanic islands provide exceptional models with which to examine patterns of dispersal, isolation and diversification, from incipient speciation to species level radiations. Here, we investigate recent differentiation and microevolutionary change in Berthelot's pipit (Anthus berthelotii), an endemic bird species inhabiting three Atlantic archipelagos. Mitochondrial DNA sequence data and microsatellite markers were used to deduce probable colonization pathway, genetic differentiation, and gene flow among the 12 island populations. Phenotypic differentiation was investigated based on eight biologically important morphological traits. We found little mitochondrial DNA variability, with only one and four haplotypes for the control region and cytochrome b, respectively. However, microsatellite data indicated moderate population differentiation (FST=0.069) between the three archipelagos that were identified as genetically distinct units with limited gene flow. Both results, combined with the estimated time of divergence (2.5 millions years ago) from the Anthus campestris (the sister species), suggest that this species has only recently dispersed throughout these islands. The genetic relationships, patterns of allelic richness and exclusive alleles among populations suggest the species originally colonized the Canary Islands and only later spread from there to the Madeiran archipelago and Selvagen Islands. Differentiation has also occurred within archipelagos, although to a lesser degree. Gene flow was observed more among the eastern and central islands of the Canaries than between these and the western islands or the Madeiran Islands. Morphological differences were also more important between than within archipelagos. Concordance between morphological and genetic differentiation provided ambiguous results suggesting that genetic drift alone was not sufficient to explain phenotypic differentiation. The observed genetic and morphological differences may therefore be the result of differing patterns of selection pressures between populations, with Berthelot's pipit undergoing a process of incipient differentiation. PMID- 17908211 TI - Population structure and gene flow reversals in Atlantic salmon (Salmo salar) over contemporary and long-term temporal scales: effects of population size and life history. AB - Metapopulation dynamics are increasingly invoked in management and conservation of endangered species. In this context, asymmetrical gene flow patterns can be density dependent, with migration occurring mainly from larger into smaller populations, which may depend on it for their persistence. Using genetic markers, such patterns have recently been documented for various organisms including salmonids, suggesting this may be a more general pattern. However, metapopulation theory does not restrict gene flow asymmetry to 'source-sink' structures, nor need these patterns be constant over longer evolutionary timescales. In anadromous salmonids, gene flow can be expected to be shaped by various selective pressures underlying homing and dispersal ('straying') behaviours. The relative importance of these selective forces will vary spatially and for populations of different census size. Furthermore, the consequences of life-history variation among populations for dispersal and hence gene flow remain poorly quantified. We examine population structure and connectivity in Atlantic salmon (Salmo salar L.) from Newfoundland and Labrador, a region where populations of this species are relatively pristine. Using genetic variation at 13 microsatellite loci from samples (N=1346) collected from a total of 20 rivers, we examine connectivity at several regional and temporal scales and test the hypothesis that the predominant direction of gene flow is from large into small populations. We reject this hypothesis and find that the directionality of migration is affected by the temporal scale over which gene flow is assessed. Whereas large populations tend to function as sources of dispersal over contemporary timescales, such patterns are often changed and even reversed over evolutionary, coalescent-derived timescales. These patterns of population structure furthermore vary between different regions and are compatible with demographic and life-history attributes. We find no evidence for sex-biased dispersal underlying gene flow asymmetry. Our findings caution against generalizations concerning the directionality of gene flow in Atlantic salmon and emphasize the need for detailed regional study, if such information is to be meaningfully applied in conservation and management of salmonids. PMID- 17908209 TI - LplA1-dependent utilization of host lipoyl peptides enables Listeria cytosolic growth and virulence. AB - The bacterial pathogen Listeria monocytogenes replicates within the cytosol of mammalian cells. Mechanisms by which the bacterium exploits the host cytosolic environment for essential nutrients are poorly defined. L. monocytogenes is a lipoate auxotroph and must scavenge this critical cofactor, using lipoate ligases to facilitate attachment of the lipoyl moiety to metabolic enzyme complexes. Although the L. monocytogenes genome encodes two putative lipoate ligases, LplA1 and LplA2, intracellular replication and virulence require only LplA1. Here we show that LplA1 enables utilization of host-derived lipoyl peptides by L. monocytogenes. LplA1 is dispensable for growth in the presence of free lipoate, but necessary for growth on low concentrations of mammalian lipoyl peptides. Furthermore, we demonstrate that the intracellular growth defect of the DeltalplA1 mutant is rescued by addition of exogenous lipoic acid to host cells, suggesting that L. monocytogenes dependence on LplA1 is dictated by limiting concentrations of available host lipoyl substrates. Thus, the ability of L. monocytogenes and other intracellular pathogens to efficiently use host lipoyl peptides as a source of lipoate may be a requisite adaptation for life within the mammalian cell. PMID- 17908212 TI - Diversity of fungi in hair roots of Ericaceae varies along a vegetation gradient. AB - Ericaceous dwarf shrubs including Calluna vulgaris and Vaccinium spp. occur both in open heathland communities and in forest ecosystems as understory vegetation. Ericaceous shrubs were once thought to form ericoid mycorrhizal associations with a relatively narrow range of ascomycetous fungi closely related to, and including, Rhizoscyphus ericae. However, perceptions have recently changed since the realization that a broader range of ascomycete fungi, and in some cases basidiomycete fungi, can also form associations with the roots of ericaceous plants. We used a combination of molecular approaches, including denaturing gradient gel electrophoresis, terminal restriction fragment length polymorphism, cloning and sequencing, to investigate the diversity of fungi associated with C. vulgaris roots collected across a heathland/native Scots pine forest vegetation gradient. We also determined differences in fungal community composition between roots of co-occurring C. vulgaris and Vaccinium myrtillus in the forest understory. Collectively, the data show that a large diversity of potentially ericoid mycorrhizal fungal taxa associate with roots of C. vulgaris and V. myrtillus, and that ascomycetes were about 2.5 times more frequent than basidiomycetes. The assemblages of fungi associated with C. vulgaris and V. myrtillus were different. In addition, the community of fungi associated with C. vulgaris hair roots was different for samples collected from the forest, open heathland and a transition zone between the two. This separation was partly, but not entirely, due to the occurrence of typical ectomycorrhizal basidiomycetes associated with the hair roots of C. vulgaris in the forest understory. These data demonstrate that forest understory ericaceous shrubs associate with a diverse range of ascomycete and basidiomycete taxa, including typical ectomycorrhizal basidiomycetes. PMID- 17908213 TI - Founding events in species invasions: genetic variation, adaptive evolution, and the role of multiple introductions. AB - Invasive species are predicted to suffer from reductions in genetic diversity during founding events, reducing adaptive potential. Integrating evidence from two literature reviews and two case studies, we address the following questions: How much genetic diversity is lost in invasions? Do multiple introductions ameliorate this loss? Is there evidence for loss of diversity in quantitative traits? Do invaders that have experienced strong bottlenecks show adaptive evolution? How do multiple introductions influence adaptation on a landscape scale? We reviewed studies of 80 species of animals, plants, and fungi that quantified nuclear molecular diversity within introduced and source populations. Overall, there were significant losses of both allelic richness and heterozygosity in introduced populations, and large gains in diversity were rare. Evidence for multiple introductions was associated with increased diversity, and allelic variation appeared to increase over long timescales (~100 years), suggesting a role for gene flow in augmenting diversity over the long-term. We then reviewed the literature on quantitative trait diversity and found that broad sense variation rarely declines in introductions, but direct comparisons of additive variance were lacking. Our studies of Hypericum canariense invasions illustrate how populations with diminished diversity may still evolve rapidly. Given the prevalence of genetic bottlenecks in successful invading populations and the potential for adaptive evolution in quantitative traits, we suggest that the disadvantages associated with founding events may have been overstated. However, our work on the successful invader Verbascum thapsus illustrates how multiple introductions may take time to commingle, instead persisting as a 'mosaic of maladaptation' where traits are not distributed in a pattern consistent with adaptation. We conclude that management limiting gene flow among introduced populations may reduce adaptive potential but is unlikely to prevent expansion or the evolution of novel invasive behaviour. PMID- 17908214 TI - High variation and strong phylogeographic pattern among cpDNA haplotypes in Taxus wallichiana (Taxaceae) in China and North Vietnam. AB - We studied the phylogeography of Chinese yew (Taxus wallichiana), a tree species distributed over most of southern China and adjacent regions. A total of 1235 individuals from 50 populations from China and North Vietnam were analysed for chloroplast DNA variation using polymerase chain reaction-restriction fragment length polymorphism of the trnL-F intron-spacer region. A total of 19 different haplotypes were distinguished. We found a very high level of population differentiation and a strong phylogeographic pattern, suggesting low levels of recurrent gene flow among populations. Haplotype differentiation was most marked along the boundary between the Sino-Himalayan and Sino-Japanese Forest floristic subkingdoms, with only one haplotype being shared among these two subkingdoms. The Malesian and Sino-Himalayan Forest subkingdoms had five and 10 haplotypes, respectively, while the relatively large Sino-Japanese Forest subkingdom had only eight. The strong geography-haplotype correlation persisted at the regional floristic level, with most regions possessing a unique set of haplotypes, except for the central China region. Strong landscape effects were observed in the Hengduan and Dabashan mountains, where steep mountains and valleys might have been natural dispersal barriers. The molecular phylogenetic data, together with the geographic distribution of the haplotypes, suggest the existence of several localized refugia during the last glaciation from which the present-day distribution may be derived. The pattern of haplotype distribution across China and North Vietnam corresponded well with the current taxonomic delineation of the three intraspecific varieties of T. wallichiana. PMID- 17908215 TI - Testing the role of genetic factors across multiple independent invasions of the shrub Scotch broom (Cytisus scoparius). AB - Knowledge of the introduction history of invasive plants informs on theories of invasiveness and assists in the invasives management. For the highly successful invasive shrub Scotch broom, Cytisus scoparius, we analysed a combination of nuclear and chloroplast microsatellites for eight native source regions and eight independent invasion events in four countries across three continents. We found that two exotic Australian populations came from different sources, one of which was derived from multiple native populations, as was an invasive sample from California. An invasive population from New Zealand appeared to be predominantly sourced from a single population, either from the native or exotic ranges. Four invasive populations from Chile were genetically differentiated from the native range samples analysed here and so their source of introduction could not be confirmed, but high levels of differentiation between the Chilean populations suggested a combination of different sources. This extensive global data set of replicated introductions also enabled tests of key theories of invasiveness in relation to genetic diversity. We conclude that invasive populations have similar levels of high genetic diversity to native ranges; levels of admixture may vary across invasive populations so admixture does not appear to have been an essential requirement for invasion; invasive and native populations exhibit similar level of genetic structure indicating similar gene flow dynamics for both types of populations. High levels of diversity and multiple source populations for invasive populations observed here discount founder effects or drift as likely explanations for previously observed seed size differences between ranges. The high levels of genetic diversity, differential and source admixture identified for most exotic populations are likely to limit the ability to source biocontrol agents from the native region of origin of invasive populations. PMID- 17908216 TI - As the raven flies: using genetic data to infer the history of invasive common raven (Corvus corax) populations in the Mojave Desert. AB - Common raven (Corvus corax) populations in Mojave Desert regions of southern California and Nevada have increased dramatically over the past five decades. This growth has been attributed to increased human development in the region, as ravens have a commensal relationship with humans and feed extensively at landfills and on road-killed wildlife. Ravens, as a partially subsidized predator, also represent a problem for native desert wildlife, in particular threatened desert tortoises (Gopherus agassizii). However, it is unclear whether the more than 15-fold population increase is due to in situ population growth or to immigration from adjacent regions where ravens have been historically common. Ravens were sampled for genetic analysis at several local sites within five major areas: the West Mojave Desert (California), East Mojave Desert (southern Nevada), southern coastal California, northern coastal California (Bay Area), and northern Nevada (Great Basin). Analyses of mtDNA control region sequences reveal an increased frequency of raven 'Holarctic clade' haplotypes from south to north inland, with 'California clade' haplotypes nearly fixed in the California populations. There was significant structuring among regions for mtDNA, with high F(ST) values among sampling regions, especially between the Nevada and California samples. Analyses of eight microsatellite loci reveal a mostly similar pattern of regional population structure, with considerably smaller, but mostly significant, values. The greater mtDNA divergences may be due to lower female dispersal relative to males, lower N(e), or effects of high mutation rates on maximal values of F(ST). Analyses indicate recent population growth in the West Mojave Desert and a bottleneck in the northern California populations. While we cannot rule out in situ population growth as a factor, patterns of movement inferred from our data suggest that the increase in raven populations in the West Mojave Desert resulted from movements from southern California and the Central Valley. Ravens in the East Mojave Desert are more similar to ones from northern Nevada, indicating movement between those regions. If this interpretation of high gene flow into the Mojave Desert is correct, then efforts to manage raven numbers by local control may not be optimally effective. PMID- 17908217 TI - Genetic analysis of potential postglacial watershed crossings in Central Europe by the bullhead (Cottus gobio L.). AB - Natural colonizations across watersheds have been frequently proposed to explain the present distributions of many freshwater fish species. However, detailed studies of such potential watershed crossings are still missing. Here, we investigated potential postglacial watershed crossings of the widely distributed European bullhead (Cottus gobio L.) in two different areas along the Rhine-Rhone watershed using detailed genetic analysis. The main advantage of studying bullheads vs. other freshwater fish species is that their distribution has been lightly influenced by human activities and as such, interpretations of colonization history are not confounded by artificial transplantations. The genetic analyses of eight microsatellite loci revealed strong genetic similarities between populations of both sides of the Rhine-Rhone watershed in the Lake Geneva area, giving strong evidence for a natural watershed crossing of bullheads from the upper Rhine drainage into the Rhone drainage in the Lake Geneva area likely facilitated by the retreat of the glaciers after the last glacial maximum some 20,000 years ago. Populations from the Lake Geneva basin were genetically more similar to populations from across the watershed in the upper Rhine drainage than to populations further downstream in the lower Rhone. In contrast, populations from Belfort, an area, which was not covered by ice during the last glacial maximum, showed strong genetic differentiation between populations of the upper Rhine and Rhone drainages. Based on our results on the bullhead, we propose that glacial retreat may have eased the dispersal of numerous European freshwater fish species across several geological boundaries. PMID- 17908218 TI - Genetic variation and phylogeography of free-living mouse species (genus Mus) in the Balkans and the Middle East. AB - This work presents a study of the distribution and pattern of variation throughout the ranges of three free-living mouse species of the genus Mus-M. macedonicus, M. spicilegus, and a M. cypriacus - based on sequencing of two segments of the mitochondrial DNA (mtDNA) control region. The study shows a similar level of variability in the three species and suggests their recent population expansion. The highest proportion of variation is found within populations indicating low genetic structuring. Phylogenetic analysis confirms the significant divergence of a mitochondrial lineage of M. macedonicus from Israel, recently described as a new subspecies, M. macedonicus spretoides. Conversely, no genetic hiatus is revealed between European and Asian populations of M. macedonicus macedonicus. Although phylogenetic relationships among M. spicilegus populations could not be unravelled precisely, the results suggest a recent westward expansion of the species. The mtDNA divergence between M. macedonicus and M. spicilegus is 7.3%, suggesting their split between c. 700,000 and 1 million years ago. These dates correspond with a coalescent estimate about 720,000 years ago. On the other hand, M. cypriacus appeared almost twice as divergent from the former species (4.5%) as from the latter (8.8%) suggesting a divergence of c. 430,000-610,000 years ago (coalescent approximately 490,000 years ago) and 830,000-1.2 million years ago (coalescent approximately 780,000 years ago), respectively. Approximate times of population expansion have also been estimated for all taxa and groups of populations. Existence of several glacial refuges and various colonization scenarios are discussed; since all estimated divergence times fall within interglacial periods it seems that climatic oscillations did not play a crucial role in the evolution of the three species. PMID- 17908219 TI - Phylogeography of Iris missouriensis (Iridaceae) based on nuclear and chloroplast markers. AB - We investigated the phylogeography of Iris missouriensis (Iridaceae), which is widely distributed in western North America. We utilized transposon display and DNA sequencing to quantify nuclear and chloroplast genetic structure. Our objectives were (i) to characterize the geographic structure of genetic variation throughout the species range, (ii) to test whether both margins of the range show reduced genetic diversity as predicted by north-south expansion and contraction associated with climate change, and (iii) to determine whether the subspecies Iris missouriensis ssp. longipetala is genetically distinct. We found that genetic diversity was significantly lower in the northern part of the range but was not significantly different between the central and southern regions, indicating greater stability of the southern margin vs. the northern. Among population differentiation was high (PhiPT=0.52). The largest divisions in each marker set were concordant and separated the southern Rocky Mountains and Basin and Range provinces from the remainder of the range. The boundaries of this phylogeographic break do not coincide with gaps in present-day distributions or phylogeographic breaks identified in other species, and may indicate a measure of reproductive isolation. Consistent with current treatments, we did not find support for the taxonomic placement of I. missourienis ssp. longipetala as a distinct species. Although transposon display has been used to investigate relationships among crop accessions and their wild relatives, to our knowledge, this is the first use of these markers for population-level phylogeography of a nonmodel species and further demonstrates their utility in species recalcitrant to amplified fragment length polymorphism protocols. PMID- 17908220 TI - The role of tropical dry forest as a long-term barrier to dispersal: a comparative phylogeographical analysis of dry forest tolerant and intolerant frogs. AB - We used a comparative phylogeographical approach to investigate the origins of the disjunct wet forest biota of the Golfo Dulce region along the Pacific slope of Costa Rica. This region is isolated by Pacific dry forests north and south and isolated from Caribbean wet forests by mountains. We studied three sympatric lowland frog species in the Craugastor fitzingeri species group that prefer wet forest but differ in their response to dry habitats. In dry forest, C. fitzingeri can survive along streams while C. crassidigitus and C. talamancae are entirely absent. We collected samples from across the ranges of all three species, and obtained mitochondrial DNA sequence data from the COI and cytochrome b genes. We observed significant phylogeographical structure in C. crassidigitus and C. talamancae, but much less in C. fitzingeri, demonstrating that mountain barriers and dry forest habitat have reduced mitochondrial gene flow in the strictly wet forest species. Additionally, we discovered that the Golfo Dulce and Central Panama populations of C. crassidigitus appear to have diverged in the Pliocene or earlier, suggesting that the dry forest separating these populations is old. Our phylogenetic analysis of 12 of approximately 16 species of the C. fitzingeri species group suggests that the three lowland species are each other's closest relatives. Because of this shared phylogenetic history, we attribute the striking differences in phylogeographical structure to the different ecologies of the frogs. In summary, we find that what appear to be minor differences in the natural history of these three closely related species may profoundly impact the potential for dispersal, range size, and cladogenesis. PMID- 17908221 TI - Eco-evolutionary vs. habitat contributions to invasion in salmon: experimental evaluation in the wild. AB - Although trait evolution over contemporary timescales is well documented, its influence on ecological dynamics in the wild has received much less attention particularly compared to traditional ecological and environmental factors. For example, evolution over ecologically relevant timescales is expected in populations that colonize new habitats, where it should theoretically enhance fitness, associated vital rates of survival and reproduction, and population growth potential. Nonetheless, success of exotic species is much more commonly attributed to ecological aspects of habitat quality and 'escape from enemies' in the invaded range. Here, we consider contemporary evolution of vital rates in introduced Chinook salmon (Oncorhynchus tshawytscha) that quickly colonized New Zealand and diverged over c. 26 generations. By using experimental translocations, we partitioned the roles of evolution and habitat quality in modifying geographical patterns of vital rates. Variation in habitat quality within the new range had the greatest influence on broad geographical patterns of vital rates, but locally adapted salmon still exhibited more than double the vital rate performance, and hence fitness, of nonlocal counterparts. The scope of this fitness evolution far exceeds the scale of divergence in trait values for these populations, or even the expected fitness effects of particular traits. These results suggest that contemporary evolution can be an important part of the eco-evolutionary dynamics of invasions and highlight the need for studies of the emergent fitness and ecological consequences of such evolution, rather than just changes in trait values. PMID- 17908223 TI - Salmonella enterica phage-resistant mutant colonies display an unusual phenotype in the presence of phage Felix 01. AB - AIMS: To investigate irregular colony morphology formation in Salmonella enterica serovar Typhimurium DPC6046 in the presence of a lytic phage, Felix 01. METHODS AND RESULTS: Phage-resistant derivatives of the parent strain DPC6046 were isolated which exhibited an irregular colony morphology. These were subjected to viability studies by using confocal scanning laser microscopy and live/dead BacLight stain to evaluate the cell viability within the colony. The phenomenon was also observed with other S. enterica serotypes tested which were normally sensitive to phage Felix. In the case of strain DPC6046, dead cells were clearly evident at the irregular edges of the phage-resistant colonies in locations where the cell density was lower. This colony morphology was not apparent with two other Salmonella phages tested. CONCLUSIONS: These findings support the hypothesis that the unusual morphology is due to reversion to phage sensitivity and consequent cell death within the colony as it forms. SIGNIFICANCE AND IMPACT OF THE STUDY: The irregular colony morphology observed is peculiar to phage Felix. The confocal scanning laser microscopy methodology allowed the basis for the irregular morphology to be elucidated. PMID- 17908222 TI - Pillars of Hercules: is the Atlantic-Mediterranean transition a phylogeographical break? AB - The geological history of the Mediterranean Sea, its hydrography and connection with the Atlantic Ocean have been well documented. Despite a wealth of historical and oceanographic data, the Atlantic-Mediterranean transition remains controversial at the biological level as there are discordant results regarding the biogeographical separation between the Atlantic and Mediterranean biota. The opening of the Strait of Gibraltar at the end of the Messinian Salinity Crisis (some 5.33 million years ago), removed the land barrier that impeded the marine biota allowing it to disperse freely into the Mediterranean Sea. However, present day genetic patterns suggest a limitation to gene flow for some marine species, preventing population admixture. In the last few years, a large number of studies have challenged the hypothesis of the Strait of Gibraltar representing a phylogeographical break. A review of more than 70 papers reveals no obvious relationship between either dispersal ability or life history, and observed patterns of partial or complete genetic isolation between Atlantic and Mediterranean populations. We re-analysed a selection of this large body of data (20 studies in total) in order to provide a homogeneous and coherent view on the generality of the phylogeographical patterns and the presence of a phylogeographical barrier. This offered the opportunity to summarize the state of the art on this matter and reach some general conclusions on the evolutionary history across the Atlantic-Mediterranean range. Geographically, some species in the transition zone showed step changes of allele frequencies associated with the Almeria-Oran Front rather than with the Strait of Gibraltar itself. A major part of the data describe evolutionary events well within the time frame of the Quaternary age as very few taxa pre-date closure of the Tethys Sea. Results point to a combined signature of vicariance, palaeoclimate fluctuation and life-history traits on the Atlantic-Mediterranean phylogeographical patterns. Principal component analysis failed to show any particular association between biological traits and genetic variables. It would argue that organismal determinism may play a far less significant role than marine biogeographers have generally believed. PMID- 17908224 TI - Characterization of a moderate thermophilic Nocardia species able to grow on polycyclic aromatic hydrocarbons. AB - AIMS: Our goal was the characterization of a new moderate thermophilic polycyclic aromatic hydrocarbon (PAH)-utilizing Nocardia strain. METHODS AND RESULTS: A thermophilic bacterium, strain TSH1, was isolated from a contaminated soil. The macroscopic and microscopic features fit well with the description of Nocardia species. The results of 16S rRNA gene analysis showed 100% match to the type strain of N. otitidiscaviarum DSM 43242(T). Strain TSH1 showed the same mycolic acid pattern as the type strain of N. otitidiscaviarum but its fatty acid profile did not permit identification to the species level. The carbon utilization profile of strain TSH1 was different from N. otitidiscaviarum. The results of hydrophobicity measurements showed that PAHs-grown cells were significantly more hydrophobic than LB-grown cells. Furthermore, biosurfactant production was detected during bacterial growth on different culture media. CONCLUSIONS: Strain TSH1 is capable of growing on a range of PAHs. When grown in PAHs-supplemented media, strain TSH1 showed a high affinity for the organic phase, suggesting that it can develop a hydrophobic surface. SIGNIFICANCE AND IMPACT OF THE STUDY: High cell surface hydrophobicity and capability of strain TSH1 to degrade different PAHs at 50 degrees C may make it an ideal candidate to treat PAH-contaminated desert soils. PMID- 17908225 TI - Membrane filtration method for enumeration and isolation of Alicyclobacillus spp. from apple juice. AB - AIM: To evaluate the applicability of filtration membranes for detecting Alicyclobacillus spp. spores in apple juice. METHODS AND RESULTS: Ten types of nitrocellulose membrane filters from five manufacturers were used to collect and enumerate five Alicyclobacillus spore isolates and results were compared to conventional K agar plating. Spore recovery differed among filters with an average recovery rate of 126.2%. Recovery levels also differed among spore isolates. Although significant difference (P < 0.05) in spore sizes existed, no correlation could be determined between spore size and membrane filter recovery rate. CONCLUSIONS: Recovery of spores using membrane filtration is dependent on the manufacturer and filter pore size. Correlations between spore recovery rate and spore size could not be determined. SIGNIFICANCE AND IMPACT OF THE STUDY: Low numbers of Alicyclobacillus spores in juice can be effectively detected using membrane filtration although recovery rate differences exist among different manufacturers. Use of membrane filtration is a simple, fast alternative to the week-long enrichment procedures currently employed in most quality assurance tests. PMID- 17908226 TI - Isolation of Escherichia coli O157:H7 strains that do not produce Shiga toxin from bovine, avian and environmental sources. AB - AIMS: To determine the potential for naturally occurring Shiga toxin-negative Escherichia coli O157 to acquire stx(2) genes. METHODS AND RESULTS: Multiple E. coli O157:H7 isolates positive for eae and ehxA, but not for stx genes, were isolated from cattle, water trough sediment, animal bedding and wild bird sources on several Ohio dairy farms. These isolates were experimentally lysogenized by stx(2)-converting bacteriophage. CONCLUSIONS: Shiga toxin-negative strains of E. coli O157 are present in multiple animal and environmental sources. SIGNIFICANCE AND IMPACT OF THE STUDY: Shiga toxin-negative strains of E. coli O157 present in the food production environment are able to acquire the stx genes, demonstrating their potential to emerge as new Shiga toxin-producing E. coli strains. PMID- 17908227 TI - Evidence for the classification of a crayfish pathogen as a member of the genus Coxiella. AB - AIMS: The study aimed to provide characterization of a potential new species of Coxiella, identified following a series of outbreaks of disease in Australian native freshwater crayfish. METHODS AND RESULTS: PCR primers designed for amplification of Coxiella burnetii genes including 16S rDNA, com1 and sodB were used to amplify homologues in the Coxiella-like crayfish pathogen. Products were then cloned and sequenced. The organism demonstrated a high degree of sequence homology in the highly conserved 16S rDNA (96%) and sodB (99%) genes, as well as the Coxiella sp. specific com1 (100%) gene. Regions flanking the sodB coding sequence demonstrated homology to C. burnetii antioxidant AhpC/Tsa family protein and dihydrodipicolinate reductase gene. CONCLUSIONS: The degree of homology between the genes selected and flanking regions suggested the two organisms were sufficiently closely related to belong to the same genus. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provided evidence for a potential new species in the currently monospecific genus Coxiella, with the only described member being C. burnetii, a category B biological warfare agent. PMID- 17908228 TI - Tetanus toxin production in soy-based medium: nutritional studies and scale-up into small fermentors. AB - AIMS: To further improve the soy-based medium, devoid of animal and dairy products, for a production of tetanus toxin by nutritional studies and to scale up the Clostridium tetani process into small fermentors. METHODS AND RESULTS: Optimum production of tetanus toxin did not require addition of pantothenic acid, thiamine, riboflavin, pyridoxine, biotin and uracil, growth factors used by previous investigators. Furthermore, l-tyrosine and l-cysteine could be eliminated from our soy-based medium without effect. Seven carbon sources were compared with glucose in the soy-based medium, but none was found to be superior to glucose. The process was successfully scaled-up into 250-ml bottles, 1-l bottles and 1-l fermentors. CONCLUSIONS: Quite remarkably, when comparing the tetanus production process in our soy-based medium with the traditional animal/dairy-containing media, our medium does not require addition of expensive vitamins, uracil or carbon sources other than glucose. Furthermore, the l tyrosine and l-cysteine components could be eliminated, making the medium (Hy Soy, glucose, powdered iron and inorganic salts) much more simple and economical. The successful scale-up from test tubes into 1-l fermentors allows us to predict that further scale-up into large fermentors will be successful. SIGNIFICANCE AND IMPACT OF THE STUDY: Toxoid preparations made from toxin produced with animal and dairy products can contain undesirable contaminants such as the prion causing bovine spongiform encephalopathy (BSE; mad cow's disease) or antigenic peptides that stimulate anaphylactic reactions and other undesirable immune reactions in immunized hosts. Our vegetable-based process avoids such unfortunate possibilities. The medium, having been made simpler and less expensive, and shown to be scaleable from test tubes into small fermentors, should be excellent for large scale production of tetanus toxin. PMID- 17908229 TI - On the compositing of samples for qualitative microbiological testing. AB - The introduction of legislative microbiological criteria for foods [Official J L338 (2005) 1] has increased the exposure of enforcement and industrial laboratories to the need to test multiple food samples for organisms such as salmonellae. A consequence has been an increase in the number of organizations, both official and commercial, considering whether it is permissible to composite replicate sample units for the purposes of assessing compliance with the criteria. This note summarizes the statistical and practical aspects of compositing sample units for compliance testing. Provided that the method of choice is 'fit for purpose', then there is no statistical difference between testing, say, 30 x 25 g sample units or 3 x 250 g sample units. However, compositing of sample units should be done only when the method has been demonstrated unequivocally to be sufficiently sensitive to detect potentially lower numbers of target organism(s) in the quantity of composited sample under the conditions of test. Different approaches to compositing of samples are considered. PMID- 17908230 TI - Rapid construction of Campylobacter jejuni deletion mutants. AB - AIMS: To develop a novel method for rapid construction of Campylobacter jejuni deletion mutants. METHODS AND RESULTS: We used overlapping extension PCR protocol to amplify a target sequence region of Camp. jejuni genomic DNA in which an internal fragment, Cj0618 coding sequence, was replaced by a chloramphenicol resistance cassette. After the resulting PCR product was introduced into electrocompetent Camp. jejuni 81-176, chloramphenicol-resistant mutants in which the wild type allele has been replaced by the deletion cassette were selected. DNA sequencing confirmed precise deletion in the Cj0618 gene. As expected from the previously reported role of Cj0618 in chick colonization, the resulting deletion mutant showed a caecal colonization defect in chick infection. CONCLUSIONS: This method can be used for rapid construction of Camp. jejuni deletion mutants. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of this method should facilitate functional characterization of various Camp. jejuni genes. PMID- 17908231 TI - Detection of Helicobacter-like DNA in the gastric mucosa of Thoroughbred horses. AB - AIMS: To assess the presence of Helicobacter DNA in the gastric mucosa Thoroughbred horses. METHODS AND RESULTS: Squamous and glandular mucosa samples were collected from 20 Thoroughbreds. None of these horses had shown any clinical symptoms of gastrointestinal disease. Necropsy tissues were analysed using histopathological techniques and a Helicobacter genus-specific PCR assay followed by sequencing of the amplicons. Seven horses were diagnosed with gastric ulceration, five with gastritis and six with both pathologies. Only two horses had a healthy gastric mucosa. Helicobacter-like DNA was detected in two out of seven horses with gastric ulcers, three out of five horses with gastritis, five out of six horses with both pathologies and one horse with normal gastric mucosa. The sequences of 1195 and 1237 bp fragments of the 16S rRNA gene shared 99% identity with the Helicobacter pylori 16S rRNA gene. However, all the samples were negative when tested with H. pylori-specific PCR assays targeting the cagA and glmM genes. CONCLUSIONS: The Helicobacter genus might colonize the gastric mucosa of horses. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of Helicobacter-like DNA in the gastric mucosa of horses and the pathogenic potential of these organisms requires further investigation. PMID- 17908232 TI - The prevalence and causes of sexual problems among premenopausal Turkish women. AB - INTRODUCTION: Sexual dysfunction in females is an important public health problem worldwide. It is suggested that sexual problems among women are more common than the number of diagnosed female sexual dysfunction (FSD) cases indicates. AIM: To determine the frequency and causes of sexual problems among premenopausal and married women who attend primary healthcare facilities. METHODS: This study was conducted at the Mother and Child Health and Family Planning Center. All women who attended this center during a 3-month period were included in the study. Sexual problems were evaluated via questionnaire and a standardized scale known as the Golombok Rust Inventory of Sexual Satisfaction (GRISS). MAIN OUTCOME MEASURE. A cross-sectional study. RESULTS: Although a total of 422 women aged 19 51 years were eligible for inclusion in the study, the participation rate was 27%. Nearly two-thirds of the women were aged 20-34 years, and of this group, 84.3% were unemployed. According to self-reports, 15.7% (18) of the women had sexual problems, whereas the prevalence of sexual dysfunction using GRISS was 26.1%. Vaginismus (41.7%), infrequent intercourse (39.1%), and nonsensuality (38.3%) were the most common complaints of the women with sexual problems. The rate of sexual dissatisfaction was found to be 7%. Sexual problems among women who had a long-term marriage (more than 11 years) and who were sexually inexperienced at the time of their marriage were significantly higher (P = 0.036, P = 0.034, respectively). It was found that discussing sexual problems with husbands and healthcare professionals did not reduce sexual problems. CONCLUSIONS: According to GRISS, nearly one-quarter of the women were suffering from sexual problems. The most common sexual problem was vaginismus, followed by infrequent intercourse. It is suggested that inadequate knowledge and the attitudes of spouses and health workers in primary healthcare settings are the important causes of FSD in this population. PMID- 17908233 TI - Prevalence and correlates of erectile dysfunction (ED) and treatment seeking for ED in Asian Men: the Asian Men's Attitudes to Life Events and Sexuality (MALES) study. AB - INTRODUCTION: There have been limited multiregional studies in Asia examining the parameters of men's general and sexual health and quality of life in the general population vs. those in clinical cohorts of patients with erectile dysfunction (ED). AIMS: The aims of the Asian Men's Attitudes to Life Events and Sexuality (Asian MALES) study were to investigate the prevalence of ED, associated health conditions, and ED treatment-seeking patterns in the general male population in five regions of Asia (China, Japan, Korea, Malaysia, and Taiwan). MAIN OUTCOME MEASURE: Standardized questionnaire previously used in a similar multiregional study and modified to ensure culturally appropriate content for Asia. METHODS: Phase I of the study involved 10,934 adult men, aged 20-75 years, who were interviewed using the standardized questionnaire. Phase II of the study involved men with self-reported ED recruited from Phase I and via physician referral, invitations in general practitioner offices, and street interception (total Phase II sample, N = 1,209). RESULTS: The overall prevalence of self-reported ED in the Phase I study population was 6.4%. ED prevalence varied by region and significantly increased with age (P < 0.01). Men with ED reported significantly greater rates of comorbid illness (P < 0.0001) and a reduced quality of life (P = 0.0001), compared with men without ED. Phase II of the study revealed that fewer than half of men with self-reported ED had sought treatment for their problem. Men were more likely to seek help for erection difficulties from Western doctors than from traditional medicine practitioners (P = 0.0001). A man's partner/spouse was the most common influencer of treatment seeking in all regions except Malaysia. CONCLUSION: The findings confirm those of existing research on ED in both Asian and non-Asian males: ED is a prevalent condition; the prevalence of ED increases with age and is strongly associated with comorbid conditions; and the majority of men have never sought treatment for their condition. This study highlights a substantial need for the evaluation and treatment of ED in Asian men. PMID- 17908234 TI - Is the current practice providing an integrated approach to the management of LUTS and ED in primary care? An audit and literature review. AB - INTRODUCTION: Lower urinary tract symptoms (LUTS) and erectile dysfunction (ED) are highly prevalent in aging men. A common pathophysiology is hypothesized to explain causal link. However, prevalence of ED in patients with LUTS remains underdiagnosed, as we believe general practitioners (GPs) do not inquire about ED in men presenting with LUTS. AIM: Our goal to find out if LUTS and ED were dealt with in an integrated fashion in primary care. METHODS: One hundred consecutive patients with LUTS attending the prostate assessment clinic anonymously completed a locally developed, qualitatively validated questionnaire and sexual health inventory for men. MAIN OUTCOME MEASURES: (i) Prevalence of concomitant ED in men presenting with LUTS; (ii) proportion of GPs enquiring about ED; (iii) patients with LUTS + ED who were offered treatment in primary care; and (iv) patients who sought treatment on review in secondary care. RESULTS: The age of patients was 39 86 years. Fifty-four percent admitted to ED: 66% >/=60 years, and 28% 14 days in vitro), synaptically-active hippocampal neurons in culture. Using a protocol where neurons are exposed to an EC(50) concentration of the muscarinic agonist methacholine (MCh) prior to (R1), and following (R2) a desensitizing pulse of a high concentration of this agonist, we have found that the reduction in M(1) mACh receptor responsiveness is decreased in quiescent (+tetrodotoxin) neurons and increased when synaptic activity is enhanced by blocking GABA(A) receptors with picrotoxin. The picrotoxin-mediated effect on M1 mACh receptor responsiveness was completely prevented by alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid receptor blockade. Inhibition of endogenous G protein-coupled receptor kinase 2 by transfection with the non-G(q/11)alpha binding, catalytically-inactive (D110A,K220R)G protein-coupled receptor kinase 2 mutant, decreased the extent of M1 mACh receptor desensitization under all conditions. Pharmacological inhibition of protein kinase C (PKC) activity, or chronic phorbol ester-induced PKC down-regulation had no effect on agonist mediated receptor desensitization in quiescent or spontaneously synaptically active neurons, but significantly decreased the extent of receptor desensitization in picrotoxin-treated neurons. MCh stimulated the translocation of diacylglycerol- sensitive eGFP-PKCepsilon, but not Ca2+/diacylglycerol sensitive eGFP-PKCbetaII in both the absence, and presence of tetrodotoxin. Under these conditions, MCh-stimulated eGFP-myristoylated, alanine-rich C kinase substrate translocation was dependent on PKC activity, but not Ca2+/calmodulin. In contrast, picrotoxin-driven translocation of myristoylated, alanine-rich C kinase substrate was accompanied by translocation of PKCbetaII, but not PKCepsilon, and was dependent on PKC and Ca2+/calmodulin. Taken together these data suggest that the level of synaptic activity may determine the different kinases recruited to regulate M1 mACh receptor desensitization in neurons. PMID- 17908241 TI - Kinase activities increase during the development of tauopathy in htau mice. AB - Hyperphosphorylated tau aggregates are the core constituent of neurofibrillary tangles. Recent research has shown a division between the presence of tangles, neurodegeneration and subsequent memory impairment, raising the possibility that an earlier pre-aggregated form of tau may be toxic. To gain further insight into the relationship between abnormal forms of tau, we have analyzed pathological changes in tau during tauopathy development in tangle-forming transgenic mice. In addition, we have quantified changes in the endogenous levels of a panel of protein kinases. We show progressive increases in aggregated tau and disease specific conformational change, with hyperphosphorylation occurring in an age dependent manner at specific sites. There were significant correlations between specific phosphorylation changes and amounts of aggregated tau and and abnormal tau conformations. Of the protein kinases tested, we found increases in phosphorylated (activated) p38 and the cyclin-dependent kinase-5 neuronal activators, p35 and p25, with aging, in the htau line, but not in non-tangle forming control mice. Changes in tau kinases correlated with the amount of tau present in abnormal conformations and with insoluble tau in htau mice. These data suggest that cdk5 and p38 may be associated with pathological changes in wild type human tau during the progressive development of tauopathy. PMID- 17908242 TI - Comparison of the pharmacological properties of GK11 and MK801, two NMDA receptor antagonists: towards an explanation for the lack of intrinsic neurotoxicity of GK11. AB - Over-stimulation of NMDA receptors (NMDARs) is involved in many neurodegenerative disorders. Thus, developing safe NMDAR antagonists is of high therapeutic interest. GK11 is a high affinity uncompetitive NMDAR antagonist with low intrinsic neurotoxicity, shown to be promising for treating CNS trauma. In the present study, we investigated the molecular basis of its interaction with NMDARs and compared this with the reference molecule MK801. We show, on primary cultures of hippocampal neurons, that GK11 exhibits neuroprotection properties similar to those of MK801, but in contrast with MK801, GK11 is not toxic to neurons. Using patch-clamp techniques, we also show that on NR1a/NR2B receptors, GK11 totally blocks the NMDA-mediated currents but has a six-fold lower IC(50) than MK801. On NR1a/NR2A receptors, it displays similar affinity but fails to totally prevent the currents. As NR2A is preferentially localized at synapses and NR2B at extrasynaptic sites, we investigated, using calcium imaging and patch-clamp approaches, the effects of GK11 on either synaptic or extrasynaptic NMDA-mediated responses. Here we demonstrate that in contrast with MK801, GK11 better preserve the synaptic NMDA-mediated currents. Our study supports that the selectivity of GK11 for NR2B containing receptors accounts contributes, at least partially, for its safer pharmacological profile. PMID- 17908244 TI - Contrasting patterns of synonymous and nonsynonymous sequence evolution in asexual and sexual freshwater snail lineages. AB - In asexual lineages, both synonymous and nonsynonymous sequence polymorphism may be reduced due to severe founder effects when asexual lineages originate. However, mildly deleterious (nonsynonymous) mutations may accumulate after asexual lineages are formed, because the efficiency of purifying selection is reduced even in the nonrecombining mitochondrial genome. Here we examine patterns of synonymous and nonsynonymous mitochondrial sequence polymorphism in asexual and sexual lineages of the freshwater snail Campeloma. Using clade-specific estimates, we found that synonymous sequence polymorphism was significantly reduced by 75% in asexuals relative to sexuals, whereas nonsynonymous sequence polymorphism did not differ significantly between sexuals and asexuals. Two asexual clades had high negative values for Tajima's D statistic. Coalescent simulations confirmed that various bottleneck scenarios can account for this result. We also used branch-specific estimates of the ratio of amino acid to silent substitutions, K(a)/K(s). Our study revealed that K(a)/K(s) ratios are six times higher in terminal branches of independent asexual lineages compared to sexuals. Coalescent-based reconstruction of gene networks for all sexual and asexual clades indicated that nonsynonymous mutations occurred at a higher frequency in recently derived asexual haplotypes. These findings suggest that patterns of synonymous and nonsynonymous nucleotide polymorphism in asexual snail lineages may be shaped by both severe founder effect and relaxed purifying selection. PMID- 17908243 TI - Epistatic and cytonuclear interactions govern outbreeding depression in the autotetraploid Campanulastrum americanum. AB - The consequences of combining divergent genomes among populations of a diploid species often involve F1 hybrid vigor followed by hybrid breakdown in later recombinant generations. As many as 70% of plant species are thought to have polyploid origins; yet little is known about the genetic architecture of divergence in polyploids and how it may differ from diploid species. We investigated the genetic architecture of population divergence using controlled crosses among five populations of the autotetraploid herb, Campanulastrum americanum. Plants were reciprocally hybridized to produce F1, F2, and F1 backcross generations that were grown with parental types in a greenhouse and measured for performance. In contrast to diploid expectations, most F1 hybrids lacked heterosis and instead showed strong outbreeding depression for early life traits. Recombinant hybrid generations often showed a recovery of performance to levels approximating, or at times even exceeding, the parental values. This pattern was also evident for an index of cumulative fitness. Analyses of line means indicated nonadditive gene action, especially forms of digenic epistasis, often influenced hybrid performance. However, standard diploid genetic models were not adequate for describing the underlying genetic architecture in a number of cases. Differences between reciprocal hybrids indicated that cytoplasmic and/or cytonuclear interactions also contributed to divergence. An enhanced role of epistasis in population differentiation may be the norm in polyploids, which have more gene copies. This study, the first of its kind on a natural autotetraploid, suggests that gene duplication may cause polyploid populations to diverge in a fundamentally different way than diploids. PMID- 17908245 TI - The quick and the dead? Sperm competition and sexual conflict in sea. AB - Our view of sperm competition is largely shaped by game-theoretic models based on external fertilizers. External fertilization is of particular interest as it is the ancestral mode of reproduction and as such, relevant to the evolution and maintenance of anisogamy (i.e., large eggs and tiny, numerous sperm). Current game-theoretic models have been invaluable in generating predictions of male responses to sperm competition in a range of internal fertilizers but these models are less relevant to marine broadcast spawners, the most common and archetypal external fertilizers. Broadcast spawners typically have incomplete fertilization due to sperm limitation and/or polyspermy (too many sperm), but the effects of incomplete (<100% fertilization rates) fertilization on game-theoretic predictions are unclear particular with regards to polyspermy. We show that incorporating the effects of sperm concentration on fertilization success changes the predictions of a classic game-theoretic model, dramatically reversing the relationship between sperm competition and the evolutionarily stable sperm release strategy. Furthermore, our results suggest that male and female broadcast spawners are likely to be in conflict at both ends of the sperm environment continuum rather than only in conditions of excess sperm as previously thought. Across the majority of the parameter space we explored, males release either too little to too much sperm for females to achieve complete fertilization. This conflict could result in a coevolutionary race that may have led to the evolution of internal fertilization in marine organisms. PMID- 17908246 TI - Patterns of reproductive isolation in Mediterranean deceptive orchids. AB - The evolution of reproductive isolation is of central interest in evolutionary biology. In plants, this is typically achieved by a combination of pre- and postpollination mechanisms that prevent, or limit, the amount of interspecific gene flow. Here, we investigated and compared two ecologically defined groups of Mediterranean orchids that differ in pollination biology and pollinator specificity: sexually deceptive orchids versus food-deceptive orchids. We used experimental crosses to assess the strength of postmating prezygotic, and postzygotic reproductive isolation, and a phylogenetic framework to determine their relative rates of evolution. We found quantitative and qualitative differences between the two groups. Food-deceptive orchids have weak premating isolation but strong postmating isolation, whereas the converse situation characterizes sexually deceptive orchids. Only postzygotic reproductive isolation among food-deceptive orchids was found to evolve in a clock-like manner. Comparison of evolutionary rates, within a common interval of genetic distance, showed that the contribution of postmating barriers was more relevant in the food deceptive species than in the sexually deceptive species. Asymmetry in prezygotic isolation was found among food-deceptive species. Our results indicate that postmating barriers are most important for reproductive isolation in food deceptive orchids, whereas premating barriers are most important in sexually deceptive orchids. The different rate of evolution of reproductive isolation and the relative strength of pre- and postmating barriers may have implication for speciation processes in the two orchid groups. PMID- 17908247 TI - Adaptive population differentiation in phenology across a latitudinal gradient in European aspen (Populus tremula, L.): a comparison of neutral markers, candidate genes and phenotypic traits. AB - A correct timing of growth cessation and dormancy induction represents a critical ecological and evolutionary trade-off between survival and growth in most forest trees (Rehfeldt et al. 1999; Horvath et al. 2003; Howe et al. 2003). We have studied the deciduous tree European Aspen (Populus tremula) across a latitudinal gradient and compared genetic differentiation in phenology traits with molecular markers. Trees from 12 different areas covering 10 latitudinal degrees were cloned and planted in two common gardens. Several phenology traits showed strong genetic differentiation and clinal variation across the latitudinal gradient, with Q(ST) values generally exceeding 0.5. This is in stark contrast to genetic differentiation at several classes of genetic markers (18 neutral SSRs, 7 SSRs located close to phenology candidate genes and 50 SNPs from five phenology candidate genes) that all showed F(ST) values around 0.015. We thus find strong evidence for adaptive divergence in phenology traits across the latitudinal gradient. However, the strong population structure seen at the quantitative traits is not reflected in underlying candidate genes. This result fit theoretical expectations that suggest that genetic differentiation at candidate loci is better described by F(ST) at neutral loci rather than by Q(ST) at the quantitative traits themselves. PMID- 17908248 TI - Evidence for coevolution of sociality and relative brain size in three orders of mammals. AB - As the brain is responsible for managing an individual's behavioral response to its environment, we should expect that large relative brain size is an evolutionary response to cognitively challenging behaviors. The "social brain hypothesis" argues that maintaining group cohesion is cognitively demanding as individuals living in groups need to be able to resolve conflicts that impact on their ability to meet resource requirements. If sociality does impose cognitive demands, we expect changes in relative brain size and sociality to be coupled over evolutionary time. In this study, we analyze data on sociality and relative brain size for 206 species of ungulates, carnivores, and primates and provide, for the first time, evidence that changes in sociality and relative brain size are closely correlated over evolutionary time for all three mammalian orders. This suggests a process of coevolution and provides support for the social brain theory. However, differences between taxonomic orders in the stability of the transition between small-brained/nonsocial and large-brained/social imply that, although sociality is cognitively demanding, sociality and relative brain size can become decoupled in some cases. Carnivores seem to have been especially prone to this. PMID- 17908250 TI - Sources of genetic and phenotypic variance in fertilization rates and larval traits in a sea urchin. AB - In nonresource based mating systems females are thought to derive indirect genetic benefits by mating with high-quality males. Such benefits can be due either to the intrinsic genetic quality of sires or to beneficial interactions between maternal and paternal haplotypes. Animals with external fertilization and no parental care offer unrivaled opportunities to address these hypotheses. With these systems, cross-classified breeding designs and in vitro fertilization can be used to disentangle sources of genetic and environmental variance in offspring fitness. Here, we employ these approaches in the Australian sea urchin Heliocidaris erythrogramma and explore how sire-dam identities influence fertilization rates, embryo viability (survival to hatching), and metamorphosis, as well as the interrelationships between these potential fitness traits. We show that fertilization is influenced by a combination of strong maternal effects and intrinsic male effects. Our subsequent analysis of embryo viability, however, revealed a highly significant interaction between parental genotypes, indicating that partial incompatibilities can severely limit offspring survival at this life history stage. Importantly, we detected no significant relationship between fertilization rates and embryo viability. This finding suggests that fertilization rates should not be inferred from hatching rates, which is commonly practiced in species in which it is not possible to estimate fertilization at conception. Finally, we detected significant additive genetic variance due to sires in rates of juvenile metamorphosis, and a positive correlation between fertilization rates and metamorphosis. This latter finding indicates that the performance of a male's ejaculate in noncompetitive IVF trials predicts heritable offspring traits, although the fitness implications of variance in rates of spontaneous juvenile metamorphosis have yet to be determined. PMID- 17908251 TI - Evolution of host specificity drives reproductive isolation among RNA viruses. AB - Ecological speciation hypotheses claim that assortative mating evolves as a consequence of divergent natural selection for ecologically important traits. Reproductive isolation is expected to be particularly likely to evolve by this mechanism in species such as phytophagous insects that mate in the habitats in which they eat. We tested this expectation by monitoring the evolution of reproductive isolation in laboratory populations of an RNA virus that undergoes genetic exchange only when multiple virus genotypes coinfect the same host. We subjected four populations of the RNA bacteriophage phi6 to 150 generations of natural selection on a novel host. Although there was no direct selection acting on host range in our experiment, three of the four populations lost the ability to infect one or more alternative hosts. In the most extreme case, one of the populations evolved a host range that does not contain any of the hosts infectible by the wild-type phi6. Whole genome sequencing confirmed that the resulting reproductive isolation was due to a single nucleotide change, highlighting the ease with which an emerging RNA virus can decouple its evolutionary fate from that of its ancestor. Our results uniquely demonstrate the evolution of reproductive isolation in allopatric experimental populations. Furthermore, our data confirm the biological credibility of simple "no-gene" mechanisms of assortative mating, in which this trait arises as a pleiotropic effect of genes responsible for ecological adaptation. PMID- 17908252 TI - Gene dynamics and nuclear architecture during differentiation. AB - Recent advances have demonstrated that placing genes in a specific nuclear context plays an important role in the regulation of coordinated gene expression, thus adding an additional level of complexity to the mechanisms of gene regulation. Differentiation processes are characterized by dynamic changes in gene activation and silencing. These alterations are often accompanied by gene relocations in relation to other genomic regions or to nuclear compartments. Unraveling of mechanisms and dynamics of chromatin positioning will thus expand our knowledge about cellular differentiation. PMID- 17908253 TI - Interaction of anaesthetic drugs and UV-B irradiation in the anterior segment of the rat eye. AB - PURPOSE: To determine the impact of anaesthesia on acute transient cataractogenesis and ultraviolet radiation (UVR)-induced cataractogenesis. METHODS: Sprague-Dawley rats were anaesthetized with pentobarbital, which caused almost full eyelid closure, or xylazine/ketamine, which caused eyelid retraction and proptosis. The eyelids of one eye were kept open with either a suture or adhesive tape, or both. The other eye was kept closed with either a suture or tape. Cataract was graded clinically and quantified in vitro as intensity of forward light scattering. In two UVR experiments, anaesthetized rats were irradiated unilaterally with 5 kJ/m2 UVR-B 300 nm for 15 mins. The difference between the two UVR experiments was the degree of proptosis in the pentobarbital group. Corneal drying was judged clinically with a grading scale. RESULTS: Within 60 mins of anaesthesia induction in the first experiment, almost all lenses in open eyes developed cataract, whereas all lenses in closed eyes remained clear. In the first UVR experiment the lens light scattering was significantly higher in the xylazine/ketamine group. In the second UVR experiment the pentobarbital group was treated to achieve proptosis similar to that in the xylazine/ketamine group, which led to a smaller difference in lens light scattering between the two anaesthesia groups. Lens light scattering in the pentobarbital groups was significantly higher with forced proptosis than without prominent proptosis. CONCLUSIONS: Xylazine/ketamine anaesthesia facilitates the development of UVR induced cataract, whereas pentobarbital anaesthesia does not. Xylazine/ketamine anaesthesia induces more proptosis and therefore leads to increased exposure of the cornea and, secondarily, the lens. PMID- 17908255 TI - Giant retinal tear after pneumatic retinopexy. PMID- 17908254 TI - Comparison of triamcinolone acetonide, 11-deoxycortisol and other lipid formulae for the visualization of vitreous body in the anterior chamber after posterior capsule rupture in animal models. AB - PURPOSE: The efficacy and toxicity of triamcinolone acetonide and other lipid formulae--calcium palmitate, cholesterol and 11-deoxycortisol--in the visualization of the prolapsed vitreous body in the anterior chamber after posterior capsule rupture were investigated in animal models. METHODS: In porcine eyes, a suspension of calcium palmitate, cholesterol, triamcinolone acetonide and 11-deoxycortisol was injected into the anterior chamber after intentionally creating posterior capsule rupture. Following gentle irrigation and aspiration, the vitreous body prolapsed in the anterior chamber was removed using an anterior vitrectomy cutter. In phakic rabbit eyes, the side-effects of the reagents were assessed for biomicroscopic appearance, intraocular pressure (IOP) and corneal histology. RESULTS: The suspension of calcium palmitate, cholesterol, triamcinolone acetonide and 11-deoxycortisol was effective in the visualization of the vitreous body prolapsed in the anterior chamber after posterior capsule rupture. When cholesterol and calcium palmitate were injected into the anterior chamber, they remained there; this induced a significant increase in IOP and corneal oedema. In contrast, most of the triamcinolone acetonide and 11 deoxycortisol that was injected into the anterior chamber had disappeared a day after the injection without affecting IOP or corneal endothelial density. When injected into the intravitreous cavity, triamcinolone led to a significant increase in IOP 2 and 4 weeks after the injection. However, calcium palmitate, cholesterol and 11-deoxycortisol injected into the vitreous cavity had no effect on IOP at 4 weeks. CONCLUSION: The suspension of triamcinolone acetonide and 11 deoxycortisol was effective in visualizing the vitreous body prolapsed in the anterior chamber after posterior capsule rupture. However, the amount of the reagent must be kept to a minimum to prevent the potential risk of ocular toxicities and postoperative late-onset ocular hypertension. PMID- 17908256 TI - Soft-shell technique using Viscoat and Healon 5: a prospective, randomized comparison between a dispersive-viscoadaptive and a dispersive-cohesive soft shell technique. AB - PURPOSE: To compare the efficacy of the dispersive-viscoadaptive soft-shell technique using Viscoat and Healon 5 to the dispersive-cohesive soft-shell technique in reducing corneal endothelial cell damage during cataract surgery. METHODS: In this prospective randomized study, 207 eyes of 171 cataract patients underwent phacoemulsification using the dispersive-viscoadaptive soft-shell technique (V-group, 102 eyes) with Viscoat and Healon5 or the dispersive-cohesive soft-shell technique (C-group, 105 eyes) with Viscoat and a cohesive agent (Opegan-Hi). Each group was divided into two subgroups depending on the amount of ultrasound (%Min) used during phacoemulsification. Corneal endothelial cell density was examined preoperatively and 3 months postoperatively. The endothelial cell loss was compared between the two groups, and also between the subgroups. RESULTS: The mean endothelial cell loss 3 months after surgery was 8.4 +/- 5.6% standard deviation (SD) in the V-group and 8.2 +/- 6.2% in the C-group (P = 0.787). In the subgroups with ultrasound of 10 %Min or less, the mean endothelial cell loss 3 months after surgery was 6.6 +/- 4.6% in the V-group and 5.5 +/- 5.0% in the C-group (P = 0.104). In the subgroups with ultrasound of over 10 %Min, this value was 10.6 +/- 6.3% in the V-group and 11.9 +/- 5.7% in the C-group (P = 0.413). The correlation coefficient of the endothelial cell loss rate and %Min was 0.245 (P = 0.0129) in the V-group and 0.501 (P < 0.0001) in the C-group. CONCLUSION: The dispersive-viscoadaptive soft-shell technique is as effective as the dispersive-cohesive soft-shell technique in protecting corneal endothelial cells during phacoemulsification regardless of the amount of ultrasound energy used. PMID- 17908257 TI - Resolution of macular oedema in occult choroidal neovascularization under oral Sorafenib treatment. AB - PURPOSE: To report on the effect of oral nexavar (Sorafenib) treatment in one patient with neovascular age-related macular degeneration (AMD) and advanced renal cell cancer (RCC). METHODS: After two intravitreal injections of bevacizumab (1.25 mg) for occult choroidal neovascularization (CNV) in AMD, the patient was started on oral Sorafenib (400 mg twice daily) treatment for RCC. RESULTS: Visual acuity (VA) was 20/80 in the left eye and optical coherence tomography (OCT) demonstrated persistent central thickening to 251 microm after bevacizumab. After 6 weeks of oral Sorafenib treatment, VA had increased to 20/70 and a significant decrease in retinal thickness to 208 microm was observed on OCT. The patient remained stable during a further 3 months of follow-up. CONCLUSIONS: Resolution of macular oedema and stabilization of VA under oral treatment with the multikinase inhibitor Sorafenib was observed. This observation warrants further investigation. PMID- 17908258 TI - A simplified method for nasolacrimal silicon intubation. PMID- 17908259 TI - Jamming of 25-gauge instruments in the cannula during vitrectomy for vitreous haemorrhage. AB - PURPOSE: To report the jamming of 25-gauge instruments in the cannula during vitreous surgery for non-clearing vitreous haemorrhage. METHODS: Forty-five eyes underwent vitrectomy with 25-gauge instruments for non-clearing vitreous haemorrhage (VH group). The incidence of 25-gauge instruments jamming in the cannula was determined retrospectively and compared with that in 112 eyes that underwent vitrectomy for epiretinal membrane (ERM group), also using 25-gauge instruments. RESULTS: The 25-gauge vitreous cutter or light pipe became jammed in the cannula in three eyes (7%) in the VH group and the instrument locked inside the cannula had to be removed with the cannula. None of the 25-gauge instruments in the ERM group jammed (p = 0.022, Fisher's exact probability test). Two of three eyes developed giant retinal breaks near the sclerotomy but no retinal break related to the sclerotomy was detected in the ERM group. Examination of the cutter revealed blood trapped between the cutter and the cannula. CONCLUSIONS: Twenty-five gauge instruments may become jammed in the cannula in eyes with non clearing vitreous haemorrhage. Clinicians should be aware of this surgical complication when 25-gauge instruments are used in vitreous haemorrhage. PMID- 17908260 TI - A population-based study of urinary symptoms and incontinence: the Canadian Urinary Bladder Survey. AB - OBJECTIVE: To ascertain the prevalence of lower urinary tract symptoms (LUTS) and urinary incontinence (UI) in Canada, using a cross-Canada telephone survey, as there is a wide discrepancy in the reported prevalence of these conditions. SUBJECTS AND METHODS: A random survey with a standardized questionnaire was conducted to elicit responses from 1000 adults, aged >or= 18 years; the sample was intended to reflect the population census. Data on age, level of education and household income were obtained from all respondents. All participants were questioned about urinary symptoms and daytime and night-time voids. For those who reported more symptoms than one episode of nocturia a more detailed questionnaire was used to ascertain symptom severity and duration. RESULTS: Data were analysed from 1000 respondents (482 men, mean age 44 years; 518 women, mean age 45 years). Half the respondents (43% of men and 57% of women) reported one or more LUTS, with nocturia the most common, at 36%. Overactive bladder (OAB) symptoms (urgency, with or with no urgency UI, usually with frequency and nocturia) were reported by 13.9% of respondents (13.1% of men and 14.7% of women). UI was reported by 28.8% of women with the 68% of these having stress UI (SUI), followed by mixed UI (MUI) in 21%, and urgency UI (UUI) in 11%. Of the 5.4% of men with UI, 27% had SUI, 15% had MUI, and 58% had UUI. Overall, the prevalence of LUTS increased with age. Respondents reported that symptoms were present for a median of 5 years. CONCLUSION: LUTS and UI are common in the Canadian population and increase with age. The prevalence in Canada of these conditions is similar to that seen in other countries. PMID- 17908261 TI - Patients with distant metastases from renal cell carcinoma can be accurately identified: external validation of a new nomogram. AB - OBJECTIVE: To identify clinical variables that can accurately predict the presence of distant metastases in patients with renal cell carcinoma (RCC). PATIENTS AND METHODS: Age, symptom classification, tumour size and the prevalence of distant metastases at diagnosis before nephrectomy were available for 5376 patients with pathologically confirmed RCC. The data of 2660 (49.5%) patients from 11 centres were used to develop a multivariable logistic regression model based nomogram predicting the individual probability of distant metastases. The remaining data from 2716 (50.5%) patients from three institutions were used for external validation. RESULTS: In the development cohort, 269/2660 (10.1%) had distant metastases, vs 285/2716 (10.5%) in the external validation cohort. Symptom classification and tumour size were independent predictors of distant metastases in the development cohort; age was not an independent predictor. A nomogram based on symptom classification and tumour size was 85.2% accurate in predicting the individual probability of distant metastases in the external validation cohort. CONCLUSION: Although distant metastases might be easily identifiable in some patients, their diagnosis might be a challenge in others. The current nomogram provides a simple, user-friendly and, most importantly, an accurate tool aimed at predicting the probability of distant metastases in patients with RCC. PMID- 17908262 TI - Evidence for a founder effect of the germline fumarate hydratase gene mutation R58P causing hereditary leiomyomatosis and renal cell cancer (HLRCC). AB - We report on the results of clinical investigation, pedigree analysis, mutation screening and haplotyping in a family with the syndrome of multiple cutaneous and uterine leiomyomas (MCUL1) and a germline missense mutation (R58P) in the fumarate hydratase gene (FH). We provide evidence for a founder effect for the identified mutation and distant relationship of our family to another familial case of MCUL1 associated with renal cell cancer, which was recently published with the same mutation. PMID- 17908263 TI - Admixture and population stratification in African Caribbean populations. AB - Throughout biomedical research, there is growing interest in the use of ancestry informative markers (AIMs) to deconstruct racial categories into useful variables. Studies on recently admixed populations have shown significant population substructure due to differences in individual ancestry; however, few studies have examined Caribbean populations. Here we used a panel of 28 AIMs to examine the genetic ancestry of 298 individuals of African descent from the Caribbean islands of Jamaica, St. Thomas and Barbados. Differences in global admixture were observed, with Barbados having the highest level of West African ancestry (89.6%+/- 2.0) and the lowest levels of European (10.2%+/- 2.2) and Native American ancestry (0.2%+/- 2.0), while Jamaica possessed the highest levels of European (12.4%+/- 3.5) and Native American ancestry (3.2%+/- 3.1). St. Thomas, USVI had ancestry levels quite similar to African Americans in continental U.S. (86.8%+/- 2.2 West African, 10.6%+/- 2.3 European, and 2.6%+/- 2.1 Native American). Significant substructure was observed in the islands of Jamaica and St. Thomas but not Barbados (K=1), indicating that differences in population substructure exist across these three Caribbean islands. These differences likely stem from diverse colonial and historical experiences, and subsequent evolutionary processes. Most importantly, these differences may have significant ramifications for case-control studies of complex disease in Caribbean populations. PMID- 17908264 TI - Two-year outcome of an intervention program for university students who have parents with alcohol problems: a randomized controlled trial. AB - BACKGROUND: Only a few intervention studies aiming to change high-risk drinking behavior have involved university students with heredity for alcohol problems. This study evaluated the effects after 2 years on drinking patterns and coping behavior of intervention programs for students with parents with alcohol problems. METHOD: In total, 82 university students (57 women and 25 men, average age 25 years) with at least 1 parent with alcohol problems were included in the study. The students were randomly assigned to 1 of the 3 programs: (i) alcohol intervention program, (ii) coping intervention program, or (iii) combination program. All the 3 intervention programs were manual based and individually implemented during 2 2-hour sessions, 4 weeks apart. Before the participants were randomly assigned, all were subjected to an individual baseline assessment. This assessment contained both a face-to-face interview and 6 self-completion questionnaires: the Alcohol Use Disorders Identification Test, estimated Blood Alcohol Concentration, Short Index of Problems, the Symptom Checklist-90, Coping with Parents' Abuse Questionnaire, and The Interview Schedule for Social Interaction (ISSI). Follow-up interviews were conducted after 1 and 2 years, respectively. The results after 1 year have previously been reported. RESULTS: All participants finished the baseline assessment, accepted and completed the intervention. Ninety-five percent of the students completed the 24-month follow up assessment. Only the group receiving the combination program continued to improve their drinking pattern significantly (p < 0.05) from the 12-month follow up to the 24-month follow-up. The improvements in this group were significantly better than in the other 2 groups. The group receiving only alcohol intervention remained at the level of improvement achieved at the 12-month follow-up. The improvements in coping behavior achieved at the 12-month follow-up remained at the 24-month follow-up for all the 3 groups, i.e., regardless of intervention program. CONCLUSION: Positive effects of alcohol intervention between 1 and 2 years were found only in the combined intervention group, contrary to the 1-year results with effects of alcohol intervention with or without a combination with coping intervention. PMID- 17908265 TI - Are racial disparities in alcohol treatment completion associated with racial differences in treatment modality entry? Comparison of outpatient treatment and residential treatment in Los Angeles County, 1998 to 2000. AB - OBJECTIVE: To determine whether racial and ethnic disparities in publicly funded alcohol treatment completion are due to racial differences in attending outpatient and residential treatment. METHODS: Statistical analysis of alcohol treatment completion rates using alcohol treatment patients' discharge records from all publicly funded treatment facilities in Los Angeles County from 1998 to 2000 (n = 10,591). RESULTS: Among these patients, African American (OR = 0.52; 95% CI 0.47, 0.57) and Hispanic (OR = 0.89; 95% CI 0.81, 0.99) patients were significantly less likely to complete treatment as compared with White patients. We found that the odds of being in outpatient versus residential care were 1.42 (95% CI 1.29, 1.55) and 2.05 (95% CI 1.85, 2.26) for African American and Hispanic alcohol treatment patients, respectively, compared with White patients. Adjusting for addiction characteristics, employment, other patient-level factors that might influence treatment enrollment, and unobserved facility-level differences through a random effects regression model, these odds increased to 1.89 (95% CI 1.22, 2.94) for African American and to 2.12 (95% CI 1.40, 3.21) for Hispanics. We developed a conditional probability model to assess the contribution of racial differences in treatment modality to racial disparities in treatment completion. Estimates from this model indicate that were African American and Hispanic patients observed in outpatient care in this population to have the same probability of receiving residential care as White patients with otherwise similar characteristics, the White-African American difference in completion rates would be reduced from 13.64% (95% CI 11.58%, 15.71%) to 11.09% (95% CI 8.77%, 13.23%) and the White-Hispanic difference would disappear, changing from 2.63% (95% CI 0.29%, 4.95%) to -0.45% (-3.52%, 2.43%). CONCLUSION: It appears that reductions in racial disparities in treatment completion could be gained by increasing enrollment in residential alcohol treatment for African American and Hispanic alcohol abusers in Los Angeles County. Further research addressing why minority alcohol abusers are less likely to receive residential alcohol treatment should be conducted, as well as research that examines why African American alcohol treatment patients have lower completion rates as compared with White patients regardless of treatment modality. PMID- 17908267 TI - Effect of DOV 102,677 on the volitional consumption of ethanol by Myers' high ethanol-preferring rat. AB - BACKGROUND: Inhibitors of monoamine neurotransmitter transporters are well established as antidepressants. However, the evidence that single (serotonin) or dual (serotonin-norepinephrine) neurotransmitter uptake inhibitors can treat ethanol abuse, either as a comorbidity with depression or as a separate entity, is inconsistent. Drugs that have, in addition, the ability to inhibit dopamine uptake may have an advantage in the treatment of alcohol abuse. Therefore, the inhibitor of norepinephrine, serotonin and dopamine uptake, DOV 102,677, was tested for its effects on the volitional consumption of ethanol by an ethanol preferring rat strain. METHODS: Myers' high ethanol-preferring rats were screened by a 10-day, 3 to 30% step-up test and then given free access to the preferred concentration of ethanol in a 3-bottle choice task. Consumption of ethanol (g/kg), water, food, and body weight were measured daily during a 3-day predrug treatment period, a 3-day treatment period, and a 3-day posttreatment period. Additional Sprague-Dawley rats were observed for 24 hours for the behavioral effects of 2.0 mg/kg s.c. reserpine after a 30-minute pretreatment with different doses of DOV 102,677. RESULTS: The triple monoamine uptake inhibitor DOV 102,677 dose-dependently decreased the volitional consumption of ethanol by as much as 71.2% (20 mg/kg i.p., b.i.d.) over 3 days of administration. This effect carried over into the posttreatment period. Similarly, the proportion of ethanol to total fluids consumed declined by 66.2% (20 mg/kg s.c., b.i.d.), while food consumption and body weight were unaltered. In contrast, amperozide (2 mg/kg i.p., b.i.d.) suppressed the amount of ethanol consumed by 56%, while naltrexone (5 mg/kg i.p., b.i.d.) was without effect. DOV 102,677 (40 mg/kg s.c.) inhibited reserpine induced akinesia and ptosis, but not hypothermia in Sprague-Dawley rats, consistent with its transient inhibition of serotonin transport, and more long lived inhibition of norepinephrine and dopamine uptake. CONCLUSIONS: DOV 102,677 significantly decreased the volitional consumption of ethanol with minimal alterations in the intake of food or on body weight in an ethanol-preferring rat strain, suggesting that triple reuptake inhibitors may find utility in treating alcohol abuse. PMID- 17908268 TI - C4d fixing, luminex binding antibodies - a new tool for prediction of graft failure after heart transplantation. AB - The standard method to detect pretransplant antibodies has been the complement dependent cytotoxicity (CDC) test of donor leukocytes. Solid phase assays to detect HLA antibodies in pretransplant serum reveal a greater number of sensitized patients, but their clinical impact is less certain. Here we have developed a method of detecting C4d fixing HLA antibodies on Luminex beads. Pretransplant serum from 565 cardiac transplant patients was retrospectively tested for the presence of HLA antibodies using CDC, HLA coated Luminex beads and C4d deposition on Luminex beads, and the results correlated with graft survival. Whereas 5/565 patients had CDC positive donor specific antibodies (DSA) before their transplant, this number was increased by 19 using Luminex beads. The 1-year survival of CDC -ve/Luminex +ve patients with DSA (n = 19) was 42% compared with 77% for CDC -ve/Luminex +ve without DSA (n = 39, p = 0.0039). Fixation of C4d (22/67 Luminex positive sera) had a negative effect on graft outcome; 1-year graft survival was, C4d +ve/DSA +ve (n = 11) 20%, C4d +ve/DSA -ve (n = 11) 91%, C4d -ve DSA +ve (n = 13) 54%, C4d -ve DSA -ve (n = 32) 75%, compared with 75% for antibody-negative patients (p = 0.0002). In conclusion, detection of Luminex +ve DSA in pretransplant serum provides a powerful negative predictor of graft survival, especially if they bind C4d. PMID- 17908266 TI - The amygdala regulates the antianxiety sensitization effect of flumazenil during repeated chronic ethanol or repeated stress. AB - BACKGROUND: The benzodiazepine receptor antagonist flumazenil reduces anxiety like behavior and sensitization of anxiety-like behavior in various models of ethanol withdrawal in rodents. The mechanism and brain region(s) that account for this action of flumazenil remain unknown. This investigation explored the potential role of several brain regions (amygdala, raphe, inferior colliculus, nucleus accumbens, and paraventricular hypothalamus) for these actions of flumazenil. METHODS: Rats were surgically implanted with guide cannulae directed over the brain region of interest and then treated with an ethanol diet for three 7-day dietary cycles (5 days on ethanol diet followed by 2 days on control diet). At approximately 4 hours, flumazenil was administered intracranially into each of the first 2 withdrawals. Examinations of anxiety-like behavior followed 1 week later during a third withdrawal. In other animals, restraint stress sessions or intra-amygdala DMCM (methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate) injections, preceded by intraperitoneal flumazenil injections, were substituted for the first 2 ethanol treatment cycles to assess the potential anxiety sensitizing action of stress or a benzodiazepine receptor inverse agonist, respectively. RESULTS: Flumazenil treatment of the amygdala during the first 2 withdrawals blocked the development of sensitized anxiety seen during a third withdrawal. Similar actions of flumazenil were found when stress sessions substituted for the first 2 cycles of ethanol exposure and withdrawal. Amygdala treatment with DMCM magnified the anxiety response to the single subthreshold chronic ethanol treatment, and prophylactic flumazenil blocked this effect. CONCLUSIONS: Intra-amygdala flumazenil inhibits the development of anxiety sensitized by repeated ethanol withdrawal, stress/ethanol withdrawal, or DMCM/ethanol withdrawal. These actions suggest that site-specific and persistent effects of flumazenil on gamma-aminobutyric acid-modulatory processes in this brain region are relevant to sensitized behavioral effects seen in alcoholism. PMID- 17908271 TI - Renal transplantation in the ANCA-associated vasculitides. AB - Despite advances in the diagnosis and treatment of the antineutrophil cytoplasmic autoantibodies (ANCA)-associated vasculitides, renal morbidity is common. End stage renal disease occurs in up to 20% of patients with these diagnoses, which include Wegener's granulomatosis and microscopic polyangiitis. As the mortality of patients with ANCA-associated vasculitis continues to improve, our ability to address the consequences of renal failure in this patient population becomes paramount. Renal transplantation is an important therapeutic option for these patients. Graft and patient survival rates among patients with ANCA-associated vasculitis are comparable to those observed in nondiabetic patients. This review summarizes our current knowledge of indications and contraindications for renal transplantation in these patients, the recurrence of vasculitis after transplantation and the impact of posttransplant immunosuppression on the clinical course of these patients. PMID- 17908273 TI - Efficacy of living donor liver transplantation for patients with methylmalonic acidemia. AB - Application of liver transplantation to methylmalonic acidemia (MMAemia) is controversial because MMAemia is caused by a systemic defect of methylmalonyl-CoA mutase. The clinical courses of seven pediatric patients with MMAemia undergoing living donor liver transplantation (LDLT) were reviewed. Serum and urinary methylmalonic acid (MMA) levels were found to be significantly decreased after LDLT, whereas serum and urinary MMA levels did not return to normal in any patient. One patient died of sepsis 44 days after LDLT. The other six patients are currently doing well. All patients had preoperative history of acute metabolic decompensation and/or metabolic stroke. However, no episode of acute metabolic decompensation or metabolic stroke was observed postoperatively in any surviving patients. In the preoperative period, all patients showed lethargy and cognitive deficit, both of which were eradicated after LDLT in all surviving patients. Preoperatively, all patients were subjected to dietary protein intake restriction and tube feeding, and were administered several metabolism-correcting medications. The metabolism-correcting medications being administered remained mostly unchanged after LDLT, whereas protein restriction was liberalized and tube feeding became unnecessary in all surviving patients. In addition, physical and neurodevelopmental growth delay remained in all surviving patients during the observation period, which ranged from 4 to 21 months with a median of 10.5 months. PMID- 17908272 TI - Alemtuzumab pre-conditioning with tacrolimus monotherapy in pediatric renal transplantation. AB - We employed antibody pre-conditioning with alemtuzumab and posttransplant immunosuppression with low-dose tacrolimus monotherapy in 26 consecutive pediatric kidney transplant recipients between January 2004 and December 2005. Mean recipient age was 10.7 +/- 5.8 years, 7.7% were undergoing retransplantation, and 3.8% were sensitized, with a PRA >20%. Mean donor age was 32.8 +/- 9.2 years. Living donors were utilized in 65% of the transplants. Mean cold ischemia time was 27.6 +/- 6.4 h. The mean number of HLA mismatches was 3.3 +/- 1.3. Mean follow-up was 25 +/- 8 months. One and 2 year patient survival was 100% and 96%. One and 2 year graft survival was 96% and 88%. Mean serum creatinine was 1.1 +/- 0.6 mg/dL, and calculated creatinine clearance was 82.3 +/ 29.4 mL/min/1.73 m(2). The incidence of pre-weaning acute rejection was 11.5%; the incidence of delayed graft function was 7.7%. Eighteen (69%) of the children were tapered to spaced tacrolimus monotherapy, 10.5 +/- 2.2 months after transplantation. The incidence of CMV, PTLD and BK virus was 0%; the incidence of posttransplant diabetes was 7.7%. Although more follow-up is clearly needed, antibody pre-conditioning with alemtuzumab and tacrolimus monotherapy may be a safe and effective regimen in pediatric renal transplantation. PMID- 17908274 TI - Successful induction of remission with rituximab for relapse of ANCA-associated vasculitis post-kidney transplant: report of two cases. AB - Kidney transplantation should be considered the treatment of choice for patients with end-stage renal disease due to antineutrophil cytoplasmic antibody (ANCA) associated vasculitis (AAV). However, relapses of AAV have been reported to occur in 9-40% of cases following kidney transplantation and may adversely affect allograft outcome. These relapses are usually treated with cyclophosphamide (CYC) and glucocorticoids, but the repeated use of CYC carries a risk of substantial toxicity that may limit or prohibit its use in some patients. B lymphocytes have been implicated in the pathogenesis of AAV, and their depletion has been effective as salvage therapy for refractory disease in the nontransplant setting. We report the successful induction of remission using rituximab in two patients who suffered relapse of AAV post-kidney transplant. Given the substantial morbidity and adverse effects of CYC, rituximab appears to be a suitable alternative agent to treat relapses of AAV posttransplantation. PMID- 17908275 TI - Prospective monitoring of polyomavirus BK replication and impact of pre-emptive intervention in pediatric kidney recipients. AB - Polyoma BK virus (BKV)-associated nephropathy (PVAN) is a relevant cause of poor renal allograft survival. In a prospective analysis, we monitored BKV DNA in blood and urine samples from 62 consecutive pediatric kidney recipients. In patients with BKV replication, we analyzed the impact of reduction of maintenance immunosuppression on viral load kinetics and PVAN in patients with BKV replication. BKV-specific immunity was concomitantly evaluated on blood samples of viremic patients, by measuring the frequency of BKV-specific interferon-gamma producing and cytotoxic T cells, and BKV IgG antibody levels. At a median follow up of 24 months, BK viruria was observed in 39 of 62 patients, while BK viremia developed in 13 patients (21%). In all viremic patients, immunosuppression reduction resulted in the clearance of viremia, and prevented development of PVAN, without increasing the rate of acute rejection or causing graft dysfunction. As a consequence of immunosuppression adjustment, an expansion of BKV-specific cellular immunity was observed that coincided with viral clearance. We conclude that treating pediatric kidney transplant patients pre-emptively with immunosuppression reduction guided by BKV DNA in blood is safe and effective to prevent onset of PVAN. BKV-specific cellular immunity may be useful to guide this intervention. PMID- 17908276 TI - Individualized mycophenolate mofetil dosing based on drug exposure significantly improves patient outcomes after renal transplantation. AB - Efficacy and safety of mycophenolate mofetil (MMF) may be optimized with individualized doses based on therapeutic monitoring of its active metabolite, mycophenolic acid (MPA). In this 12-month study, 137 renal allograft recipients from 11 French centers receiving basiliximab, cyclosporine A, MMF and corticosteroids were randomized to receive either concentration-controlled doses or fixed-dose MMF. A novel Bayesian estimator of MPA AUC based on three-point sampling was used to individualize doses on posttransplant days 7 and 14 and months 1, 3 and 6. The primary endpoint was treatment failure (death, graft loss, acute rejection and MMF discontinuation). Data from 65 patients/group were analyzed. At month 12, the concentration-controlled group had fewer treatment failures (p = 0.03) and acute rejection episodes (p = 0.01) with no differences in adverse event frequency. The MMF dose was higher in the concentration controlled group at day 14 (p < 0.0001), month 1 (p < 0.0001) and month 3 (p < 0.01), as were median AUCs on day 14 (33.7 vs. 27.1 mg*h/L; p = 0.0001) and at month 1 (45.0 vs. 30.9 mg*h/L; p < 0.0001). Therapeutic MPA monitoring using a limited sampling strategy can reduce the risk of treatment failure and acute rejection in renal allograft recipients 12 months posttransplant with no increase in adverse events. PMID- 17908277 TI - Immunogenicity and safety of an intradermal boosting strategy for vaccination against influenza in lung transplant recipients. AB - The immunogenicity of influenza vaccine is suboptimal in lung transplant recipients. Use of a booster dose and vaccine delivery by the intradermal rather than intramuscular route may improve response. We prospectively evaluated the immunogenicity and safety of a 2-dose boosting strategy of influenza vaccine. Sixty lung transplant recipients received a standard intramuscular injection of the 2006-2007 inactivated influenza vaccine, followed 4 weeks later by an intradermal booster of the same vaccine. Immunogenicity was assessed by measurement of geometric mean titer of antibodies after both the intramuscular injection and the intradermal booster. Vaccine response was defined as 4-fold or higher increase of antibody titers to at least one vaccine antigen. Thirty-eight out of 60 patients (63%) had a response after intramuscular vaccination. Geometric mean titers increased for all three vaccine antigens following the first dose (p < 0.001). However, no significant increases in titer were observed after the booster dose for all three antigens. Among nonresponders, 3/22 (13.6%) additional patients responded after the intradermal booster (p = 0.14). The use of basiliximab was associated with a positive response (p = 0.024). After a single standard dose of influenza vaccine, a booster dose given by intradermal injection did not significantly improve vaccine immunogenicity in lung transplant recipients. PMID- 17908278 TI - Plasma intercellular adhesion molecule-1 and von Willebrand factor in primary graft dysfunction after lung transplantation. AB - Primary graft dysfunction (PGD), a form of acute lung injury occurring within 72 h following lung transplantation, is characterized by pulmonary edema and diffuse alveolar damage. We hypothesized that higher concentrations of intercellular adhesion molecule-1 (ICAM-1) and von Willebrand factor (vWF) would be associated with the occurrence of PGD. A total of 128 lung transplant recipients among 7 lung transplant centers were enrolled in a multicenter, prospective, cohort study. Blood specimens were collected preoperatively and at 6, 24, 48 and 72 h following lung transplantation. The primary outcome was Grade 3 PGD at 72 h after transplant. Logistic regression and generalized estimating equations (GEE) were used to analyze plasma ICAM-1 and vWF. At each postoperative timepoint, mean plasma ICAM-1 concentrations were higher for patients with PGD versus no PGD. The GEE contrast estimate for the association of plasma ICAM-1 with PGD was 107.5 ng/mL (95% CI 38.7, 176.3), p = 0.002. In the multivariate analyses, this finding was independent of all clinical variables except pulmonary artery pressures prior to transplant. There was no association between plasma vWF levels and PGD. We conclude that higher levels of plasma ICAM-1 are associated with PGD following lung transplantation. PMID- 17908279 TI - Transcriptome analysis reveals heterogeneity in the injury response of kidney transplants. AB - We studied the transcripts that are increased by stress and injury in mouse kidney transplants, focusing on transcripts increased in parenchymal cells-injury and repair-induced transcripts (IRITs). We compared four types of stressed kidneys: isografts, allografts, host kidneys of mice with isografts and nontransplant kidneys with ischemic acute tubular necrosis (ATN). After excluding transcripts associated with infiltrating cells and interferon-gamma-induced transcripts, we defined 790 IRITs in isografts. IRITs were remarkably heterogeneous in timing and mechanisms. Some were increased in host as well as donor kidneys, reflecting systemic influences (wounding, anesthetic). Most reflected local stress, resembling changes in ATN despite the lack of ATN histopathology. Mathematical decomposition of IRIT expression patterns confirmed heterogeneity, separating IRIT changes into component subsets, with an early peak (day 1) showing systemic effects and late peaks that resembled ATN, manifested Tgf-ss1 effects and recapitulated embryonic development. In allografts IRITs were initially similar to isografts but diverged due to allogeneic injury. The allospecific induction of IRITs was T-cell-dependent but perforin-granzyme independent, compatible with delayed type hypersensitivity. The alloresponse strikingly and selectively increased the late IRITs but not the IRITs that peak early, indicating that rejection triggers parenchymal responses similar to those in ATN. PMID- 17908280 TI - Lack of benefit of early protocol biopsies in renal transplant patients receiving TAC and MMF: a randomized study. AB - We conducted a randomized, multicenter study to determine whether treatment of subclinical rejection with increased corticosteroids resulted in beneficial outcomes in renal transplant patients receiving tacrolimus (TAC), mycophenolate mofetil (MMF) and prednisone. One hundred and twenty-one patients were randomized to biopsies at 0,1,2,3 and 6 months (Biopsy arm), and 119 to biopsies at 0 and 6 months only (Control arm). The primary endpoint of the study was the prevalence of the sum of the interstitial and tubular scores (ci + ct)> 2 (Banff) at 6 months. Secondary endpoints included clinical and subclinical rejection and renal function. At 6 months, 34.8% of the Biopsy and 20.5% of the Control arm patients had a ci + ct score >or= 2 (p = 0.07). Between months 0 and 6, clinical rejection episodes were 12 in 10 Biopsy arm patients and 8 in 8 Control arm patients (p = 0.44). Overall prevalence of subclinical rejection in the Biopsy arm was 4.6%. Creatinine clearance at 6 months was 72.9 +/- 21.7 in the Biopsy and 68.90 mL/min +/- 18.35 mL/min in the Control arm patients (p = 0.18). In conclusion, we found no benefit to the procurement of early protocol biopsies in renal transplant patients receiving TAC, MMF and prednisone, at least in the short term. This is likely due to their low prevalence of subclinical rejection. PMID- 17908281 TI - Vitamin D status in renal transplant recipients. AB - Vitamin D plays an important role in calcium homeostasis. Renal transplant recipients may be more susceptible to reduced levels because of decreased sun exposure and steroid therapy. This study aimed to determine vitamin D status after renal transplantation and its effect on parathyroid hormone (PTH) and bone mineral density (BMD). We measured serum 25-hydroxyvitamin D levels (25-OHD) in 244 renal transplant recipients, divided into two groups, 104 recently transplanted (less than 1 year) and 140 long-term. Vitamin D status was defined according to NKF/KDOQI guidelines. Mean 25-OHD levels were 33 +/- 19 nmol/L and 42 +/- 20 nmol/L, respectively, for the recent and long-term transplant recipients. Vitamin D insufficiency was present in 29% and 43%, deficiency in 56% and 46% and severe deficiency in 12% and 5%, respectively. An inverse correlation was found between logPTH and 25-OHD (r=-0.2, p= 0.019) in long-term but not in recently transplanted patients. No correlation was found between 25-OHD levels and BMD. Hypercalcaemia was present in 40% of the recently transplanted recipients and 25% of the long-term. In conclusion 25-OHD was low in virtually all of our renal transplant recipients and may aggravate secondary hyperparathyroidism, but its correction may be difficult in patients with hypercalcaemia. PMID- 17908282 TI - Epidemiology of venous thromboembolism after lower limb arthroplasty: the FOTO study. AB - BACKGROUND: In view of recent substantial changes in the management of orthopedic surgery patients, a study was performed in order to update data on the epidemiology of venous thromboembolism (VTE) in patients undergoing lower limb arthroplasty according to contemporary practise. METHODS: We performed a prospective observational study of a cohort of consecutive patients hospitalized for total hip or knee replacement in June 2003. The primary study outcome was the incidence of symptomatic VTE at 3 months. All events were adjudicated by an independent critical event committee. RESULTS: Data from 1080 patients (mean age 68.0 years) were available; 63.2% were undergoing total hip replacement and 36.8% total knee replacement. Pharmacological thromboprophylaxis was administered for a mean time of 36 days. Injectable antithrombotics were used in more than 99% of patients, irrespective of the type of surgery. The incidence of the primary study outcome was 1.8% (20 events; 95% CI: 1.0-2.6%). The incidences were 1.3% and 2.8% in hip and knee surgery patients, respectively. There were two pulmonary embolisms, both in knee surgery patients; neither was fatal. Thirty-five per cent of VTEs occurred after hospital discharge. An age of at least 75 years and the absence of ambulation before hospital discharge were the only significant (P < 0.05) predictors of VTE. The rate of clinically significant bleeding was 1.0% and the rate of death was 0.9%. CONCLUSIONS: The incidence of symptomatic VTE after lower limb arthroplasty is low, even if there is still a need to improve thromboprophylaxis, notably in patients undergoing knee arthroplasty. PMID- 17908283 TI - Forward Transport of K2p3.1: mediation by 14-3-3 and COPI, modulation by p11. AB - Surface expression of the K(2P)3.1 two-pore domain potassium channel is regulated by phosphorylation-dependent binding of 14-3-3, leading to suppression of coatomer coat protein I (COPI)-mediated retention in endoplasmic reticulum (ER). Here, we investigate the nature of the macromolecular regulatory complexes that mediate forward and retrograde transport. We demonstrate that (i) the channel employs two separate but interacting COPI binding sites on the N- and C-termini; (ii) disrupting COPI binding to either site interferes with the ER retention; (iii) p11 and 14-3-3 do not interact on their own; (iv) p11 binding to the C terminal retention motif is dependent on 14-3-3; and (v) p11 is coexpressed in only a subset of tissues with K(2P)3.1, while 14-3-3 expression is ubiquitous. We conclude that K(2P)3.1 forward transport requires 14-3-3 suppression of COPI binding, whereas p11 serves a modulatory role. PMID- 17908284 TI - Endocytosis: a positive or a negative influence on Wnt signalling? AB - Endocytosis, with subsequent targeting to lysosomes for degradation, is traditionally seen as a way for cells to terminate signalling. However, in a few instances, endocytosis has been demonstrated to contribute positively to signalling. Here we review recent work on the role of endocytosis in Wnt signalling. Biochemical evidence suggests that the branch of Wnt signalling that controls planar cell polarity (PCP) does require endocytosis, although how endocytosis of Frizzled receptors is translated into PCP in vivo remains unknown. With respect to the main signalling branch (called the canonical or beta-catenin pathway), the literature is divided as to whether endocytosis is required. Results of in vivo experiments are inconclusive because of the toxic side-effects of blocking endocytosis. Some results with cultured cells suggest the need for endocytosis in canonical signalling; however, it remains unclear whether the ligand-receptor complex must enter the cell by clathrin-mediated or caveolae mediated endocytosis in order to signal. Means of specifically altering Wnt trafficking as well as of tracking the internalization route in different cell types are needed. PMID- 17908286 TI - Methods of electroencephalographic signal analysis for detection of small hidden changes. AB - The aim of this study was to select and evaluate methods sensitive to reveal small hidden changes in the electroencephalographic (EEG) signal. Two original methods were considered.Multifractal method of scaling analysis of the EEG signal based on the length distribution of low variability periods (LDLVP) was developed and adopted for EEG analysis. The LDLVP method provides a simple route to detecting the multifractal characteristics of a time-series and yields somewhat better temporal resolution than the traditional multifractal analysis.The method of modulation with further integration of energy of the recorded signal was applied for EEG analysis. This method uses integration of differences in energy of the EEG segments with and without stressor.Microwave exposure was used as an external stressor to cause hidden changes in the EEG. Both methods were evaluated on the same EEG database. Database consists of resting EEG recordings of 15 subjects without and with low-level microwave exposure (450 MHz modulated at 40 Hz, power density 0.16 mW/cm2). The significant differences between recordings with and without exposure were detected by the LDLVP method for 4 subjects (26.7%) and energy integration method for 2 subjects (13.3%).The results show that small changes in time variability or energy of the EEG signals hidden in visual inspection can be detected by the LDLVP and integration of differences methods. PMID- 17908285 TI - The effects of short-term alpha-ketoisocaproic acid supplementation on exercise performance: a randomized controlled trial. AB - BACKGROUND: This study examined the efficacy of short-term alpha-ketoisocaproic acid (KIC) monotherapy supplementation immediately prior to moderate- and high intensity single bout exercise performance. METHODS: Thirteen resistance trained men (22.8 +/- 2.5 years; 81.6 +/- 12.6 kg) participated in a prospective, randomized, double blind, placebo controlled crossover experiment. Each subject completed one familiarization and four experimental trials with either 1.5 g or 9.0 g of either KIC or isocaloric placebo control (CONT), following an overnight fast. During the experimental trials, subjects consumed the supplement regimen and then completed leg and chest press repetitions to failure and 30 s of repeated maximal vertical jumping (VJ) on a force plate. RESULTS: In this treatment regimen, no significant differences (p > 0.05) were observed between dosages or conditions for leg press (low CONT = 19.8 +/- 0.4 SEM, low KIC = 21.0 +/- 0.5, high CONT = 20.1 +/- 0.3, high KIC = 22.4 +/- 0.6) or chest press (low CONT = 18.1 +/- 0.2, low KIC = 18.5 +/- 0.3, high CONT = 17.8 +/- 0.3, high KIC = 18.0 +/- 0.3) repetitions to failure. Additionally, no significant differences were observed for peak or mean VJ performance (low CONT = 34.6 +/- 2.2 cm and 28.6 +/- 1.8 cm; low KIC = 35.6 +/- 2.0 cm and 29.4 +/- 1.6 cm; high CONT = 35.7 +/- 2.1 cm and 29.4 +/- 1.7 cm; high KIC = 34.8 +/- 2.3 cm and 28.3 +/- 1.7 cm), respectively. CONCLUSION: Based on our results, we conclude that acute KIC ingestion by itself with no other ergogenic supplement, immediately prior to exercise, did not alter moderate- nor high-intensity single-bout exercise performance in young resistance-trained males. This study addressed single-dose single-bout performance events; the efficacy of KIC monotherapy supplementation on repeated high-intensity exercise bouts and long-term exercise training remains unknown. PMID- 17908287 TI - Reconstruction of cellular variability from spatiotemporal patterns of Dictyostelium discoideum. AB - Variability in cell properties can be an important driving mechanism behind spatiotemporal patterns in biological systems, as the degree of cell-to-cell differences determines the capacity of cells to locally synchronize and, consequently, form patterns on a larger spatial scale. In principle, certain features of spatial patterns emerging with time may be regulated by variability or, more specifically, by certain constellations of cell-to-cell differences. Similarly, measuring variability in a system (i.e. the spatial distribution of cell-cell differences) may help predict properties of later-stage patterns.Here we apply and compare different statistical methods of extracting such systematic cell-to-cell differences in the case of patterns generated with a simple model system of an excitable medium and of experimental data by the slime mold Dictyostelium discoideum. We demonstrate with the help of a correlation analysis that these methods produce systematic (i.e. stationary) results for cell properties. Furthermore, we discuss possible applications of our method, in particular how these cell properties may serve as predictors of certain later stage patterns. PMID- 17908289 TI - Developing combinatorial multi-component therapies (CMCT) of drugs that are more specific and have fewer side effects than traditional one drug therapies. AB - Drugs designed for a specific target are always found to have multiple effects. Rather than hope that one bullet can be designed to hit only one target, nonlinear interactions across genomic and proteomic networks could be used to design Combinatorial Multi-Component Therapies (CMCT) that are more targeted with fewer side effects. We show here how computational approaches can be used to predict which combinations of drugs would produce the best effects. Using a nonlinear model of how the output effect depends on multiple input drugs, we show that an artificial neural network can accurately predict the effect of all 215 = 32,768 combinations of drug inputs using only the limited data of the output effect of the drugs presented one-at-a-time and pairs-at-a-time. PMID- 17908288 TI - International Society of Sports Nutrition position stand: creatine supplementation and exercise. PMID- 17908290 TI - Metabolic responses to the acute ingestion of two commercially available carbonated beverages: A pilot study. AB - BACKGROUND: The purpose of this placebo-controlled, double-blind cross-over study was to compare the effects of two commercially available soft drinks on metabolic rate. METHODS: After giving informed consent, twenty healthy men and women were randomly assigned to ingest 12 ounces of Celsiustrade mark and, on a separate day, 12 ounces of Diet Coke(R). All subjects completed both trials using a randomized, counterbalanced design. Metabolic rate (via indirect calorimetry) and substrate oxidation (via respiratory exchange ratio) were measured at baseline (pre-ingestion) and at the end of each hour for 3 hours post-ingestion. RESULTS: Two-way ANOVA revealed a significant interaction (p < 0.001) between trials in metabolic rate. Scheffe post-hoc testing indicated that metabolic rate increased by 13.8% (+ 0.6 L/min, p < 0.001) 1 hr post, 14.4% (+0.63 L/min, p < 0.001) 2 hr post, and 8.5% (+0.37 L/min, p < 0.004) 3 hr post Celsiustrade mark ingestion. In contrast, small (~4-6%) but statistically insignificant increases in metabolic rate were noted following Diet Coke(R) ingestion. No differences in respiratory exchange ratio were noted between trials. CONCLUSION: These preliminary findings indicate Celsiustrade mark has thermogenic properties when ingested acutely. The effects of repeated, chronic ingestion of Celsiustrade mark on body composition are unknown at this time. PMID- 17908291 TI - International Society of Sports Nutrition position stand: protein and exercise. AB - POSITION STATEMENT: The following seven points related to the intake of protein for healthy, exercising individuals constitute the position stand of the Society. They have been approved by the Research Committee of the Society. 1) Vast research supports the contention that individuals engaged in regular exercise training require more dietary protein than sedentary individuals. 2) Protein intakes of 1.4 - 2.0 g/kg/day for physically active individuals is not only safe, but may improve the training adaptations to exercise training. 3) When part of a balanced, nutrient-dense diet, protein intakes at this level are not detrimental to kidney function or bone metabolism in healthy, active persons. 4) While it is possible for physically active individuals to obtain their daily protein requirements through a varied, regular diet, supplemental protein in various forms are a practical way of ensuring adequate and quality protein intake for athletes. 5) Different types and quality of protein can affect amino acid bioavailability following protein supplementation. The superiority of one protein type over another in terms of optimizing recovery and/or training adaptations remains to be convincingly demonstrated. 6) Appropriately timed protein intake is an important component of an overall exercise training program, essential for proper recovery, immune function, and the growth and maintenance of lean body mass. 7) Under certain circumstances, specific amino acid supplements, such as branched-chain amino acids (BCAA's), may improve exercise performance and recovery from exercise. PMID- 17908292 TI - The complete mitochondrial genome of the sea spider Achelia bituberculata (Pycnogonida, Ammotheidae): arthropod ground pattern of gene arrangement. AB - BACKGROUND: The phylogenetic position of pycnogonids is a long-standing and controversial issue in arthropod phylogeny. This controversy has recently been rekindled by differences in the conclusions based on neuroanatomical data concerning the chelifore and the patterns of Hox expression. The mitochondrial genome of a sea spider, Nymphon gracile (Pycnogonida, Nymphonidae), was recently reported in an attempt to address this issue. However, N. gracile appears to be a long-branch taxon on the phylogenetic tree and exhibits a number of peculiar features, such as 10 tRNA translocations and even an inversion of several protein coding genes. Sequences of other pycnogonid mitochondrial genomes are needed if the position of pycnogonids is to be elucidated on this basis. RESULTS: The complete mitochondrial genome (15,474 bp) of a sea spider (Achelia bituberculata) belonging to the family Ammotheidae, which combines a number of anatomical features considered plesiomorphic with respect to other pycnogonids, was sequenced and characterized. The genome organization shows the features typical of most metazoan animal genomes (37 tightly-packed genes). The overall gene arrangement is completely identical to the arthropod ground pattern, with one exception: the position of the trnQ gene between the rrnS gene and the control region. Maximum likelihood and Bayesian inference trees inferred from the amino acid sequences of mitochondrial protein-coding genes consistently indicate that the pycnogonids (A. bituberculata and N. gracile) may be closely related to the clade of Acari and Araneae. CONCLUSION: The complete mitochondrial genome sequence of A. bituberculata (Family Ammotheidae) and the previously-reported partial sequence of Endeis spinosa show the gene arrangement patterns typical of arthropods (Limulus-like), but they differ markedly from that of N. gracile. Phylogenetic analyses based on mitochondrial protein-coding genes showed that Pycnogonida may be authentic arachnids (= aquatic arachnids) within Chelicerata sensu lato, as indicated by the name 'sea spider,' and suggest that the Cormogonida theory - that the pycnogonids are a sister group of all other arthropods - should be rejected. However, in view of the relatively weak node confidence, strand-biased nucleotide composition and long-branch attraction artifact, further more intensive studies seem necessary to resolve the exact position of the pycnogonids. PMID- 17908293 TI - Identification of genes differentially expressed during prenatal development of skeletal muscle in two pig breeds differing in muscularity. AB - BACKGROUND: Postnatal muscle growth is largely depending on the number and size of muscle fibers. The number of myofibers and to a large extent their metabolic and contractile properties, which also influence their size, are determined prenatally during the process of myogenesis. Hence identification of genes and their networks governing prenatal development of skeletal muscles will provide insight into the control of muscle growth and facilitate finding the source of its variation. So far most of the genes involved in myogenesis were identified by in vitro studies using gene targeting and transgenesis. Profiling of transcriptome changes during the myogenesis in vivo promises to obtain a more complete picture. In order to address this, we performed transcriptome profiling of prenatal skeletal muscle using differential display RT-PCR as on open system with the potential to detect novel transcripts. Seven key stages of myogenesis (days 14, 21, 35, 49, 63, 77 and 91 post conception) were studied in two breeds, Pietrain and Duroc, differing markedly in muscularity and muscle structure. RESULTS: Eighty prominent cDNA fragments were sequenced, 43 showing stage associated and 37 showing breed-associated differences in the expression, respectively. Out of the resulting 85 unique expressed sequence tags, EST, 52 could be assigned to known genes. The most frequent functional categories represented genes encoding myofibrillar proteins (8), genes involved in cell adhesion, cell-cell signaling and extracellular matrix synthesis/remodeling (8), genes regulating gene expression (8), and metabolism genes (8). Some of the EST that showed no identity to any known transcripts in the databases are located in introns of known genes and most likely represent novel exons (e.g. HMGA2). Expression of thirteen transcripts along with five reference genes was further analyzed by means of real-time quantitative PCR. Nine of the target transcripts showed higher than twofold differences in the expression between the two breeds (GATA3, HMGA2, NRAP, SMC6L1, SPP1, RAB6IP2, TJP1 and two EST). CONCLUSION: The present study revealed several genes and novel transcripts not previously associated with myogenesis and expands our knowledge of genetic factors operating during myogenesis. Genes that exhibited differences between the divergent breeds represent candidate genes for muscle growth and structure. PMID- 17908295 TI - The necessary length of carbon nanotubes required to optimize solar cells. AB - BACKGROUND: In recent years scientists have been trying both to increase the efficiency of solar cells, whilst at the same time reducing dimensions and costs. Increases in efficiency have been brought about by implanting carbon nanotubes onto the surface of solar cells in order to reduce the reflection of sunrays, as well as through the insertion of polymeric arrays into the intrinsic layer for charge separation. RESULTS: The experimental results show power rising linearly for intrinsic layer thicknesses between 0-50 nm. Wider thicknesses increase the possibility of recombination of electrons and holes, leading to perturbation of the linear behaviour of output power. This effect is studied and formulated as a function of thickness. Recognition of the critical intrinsic layer thickness can permit one to determine the length of carbon nanotube necessary for optimizing solar cells. CONCLUSION: In this study the behaviour of output power as a function of intrinsic layer thicknesses has been described physically and also simulated. In addition, the implantation of carbon nanotubes into the intrinsic layer and the necessary nanotube length required to optimize solar cells have been suggested. PMID- 17908294 TI - The frontier between cell and organelle: genome analysis of Candidatus Carsonella ruddii. AB - BACKGROUND: Bacterial symbioses are widespread among insects. The early establishment of such symbiotic associations has probably been one of the key factors for the evolutionary success of insects, since it may have allowed access to novel ecological niches and to new imbalanced food resources, such as plant sap or blood. Several genomes of bacterial endosymbionts of different insect species have been recently sequenced, and their biology has been extensively studied. Recently, the complete genome sequence of Candidatus Carsonella ruddii, considered the primary endosymbiont of the psyllid Pachpsylla venusta, has been published. This genome consists of a circular chromosome of 159,662 bp and has been proposed as the smallest bacterial endosymbiont genome known to date. RESULTS: The detailed analysis of the gene content of C. ruddii shows that the extensive degradation of the genome is not compatible with its consideration as a mutualistic endosymbiont and, even more, as a living organism. The ability to perform most essential functions for a cell to be considered alive is heavily impaired by the lack of genes involved in DNA replication, transcription and translation. Furthermore, the shortening of genes causes, in some cases, the loss of essential domains and functional residues needed to fulfill such vital functions. In addition, at least half of the pathways towards the biosynthesis of essential amino acids, its proposed symbiotic function, are completely or partially lost. CONCLUSION: We propose that this strain of C. ruddii can be viewed as a further step towards the degeneration of the former primary endosymbiont and its transformation in a subcellular new entity between living cells and organelles. Although the transition of genes from C. ruddii to the host nucleus has been proposed, the amount of genes that should have been transferred to the germinal line of the insect would be so big that it would be more plausible to consider the implication of the mitochondrial machinery encoded in the insect nucleus. Furthermore, since most genes for the biosynthesis of essential amino acids have also been lost, it is likely that the host depends on another yet unidentified symbiont to complement its deficient diet. PMID- 17908296 TI - Victimization and PTSD-like states in an Icelandic youth probability sample. AB - BACKGROUND: Although adolescence in many cases is a period of rebellion and experimentation with new behaviors and roles, the exposure of adolescents to life threatening and violent events has rarely been investigated in national probability studies using a broad range of events. METHODS: In an Icelandic national representative sample of 206 9th-grade students (mean = 14.5 years), the prevalence of 20 potentially traumatic events and negative life events was reported, along with the psychological impact of these events. RESULTS: Seventy four percent of the girls and 79 percent of the boys were exposed to at least one event. The most common events were the death of a family member, threat of violence, and traffic accidents. The estimated lifetime prevalence of posttraumatic stress disorder-like states (PTSD; DSM-IV, APA, 1994 1) was 16 percent, whereas another 12 percent reached a sub-clinical level of PTSD-like states (missing the full diagnosis with one symptom). Following exposure, girls suffered from PTSD-like states almost twice as often as boys. Gender, mothers' education, and single-parenthood were associated with specific events. The odds ratios and 95% CI for PTSD-like states given a specific event are reported. Being exposed to multiple potentially traumatic events was associated with an increase in PTSD-like states. CONCLUSION: The findings indicate substantial mental health problems in adolescents that are associated with various types of potentially traumatic exposure. PMID- 17908298 TI - Application of a zona pellucida binding assay (ZBA) in the domestic cat benefits from the use of in vitro matured oocytes. AB - BACKGROUND: Zona pellucida binding assays (ZBAs) have proven useful in determining the fertilising ability of spermatozoa in several species. Most ZBAs use fresh or salt-stored oocytes collected from fresh ovaries but because ovaries are not easy to obtain on a regular basis, chilled and frozen-thawed ovaries have been tested, with varying results. The present study tested the hypothesis that cat spermatozoa, either fresh or frozen-thawed, can bind to homologous zona pellucida of oocytes retrieved from frozen-thawed queen ovaries to a similar extent as they can bind to the zona pellucida of fresh, in vitro matured oocytes. METHODS: Ovaries were collected from queens after routine ovario-hysterectomy and either stored in NaCl at -20 degrees C until use (treatment animals), or used fresh (controls). Cumulus-oocyte complexes (COCs) were retrieved by ovarian slicing from either source and used directly (immature oocytes from frozen-thawed ovaries; treatment animals) or after in vitro maturation (IVM) (fresh ovaries; controls) for 24 hours in TCM 199, supplemented with 1 IU hCG/mL and 0.5 IU eCG/mL and 0.5% bovine serum albumin (BSA). The oocytes were incubated for 4 hours in 5% CO2 in air at 38 degrees C and 100% humidity in the presence of 5 x 106 fresh or frozen-thawed spermatozoa/mL. Representative samples of oocytes were processed for scanning electron microscopy (SEM). RESULTS: Both fresh and frozen thawed spermatozoa bound to the in vitro matured zona pellucida but significantly fewer, or no, spermatozoa bound to frozen-thawed, immature zona pellucida (P < 0.001). Also, more fresh spermatozoa than frozen-thawed spermatozoa bound to the zona pellucida (P < 0.001). The zona pellucida surface differed in morphology (SEM), with in vitro matured oocytes showing a dense surface with few fenestrations in contrast to their frozen-thawed, immature counterparts, where fenestrations were conspicuously larger. CONCLUSION: In conclusion, under the conditions of the present study, immature oocytes recovered from ovaries frozen immersed in NaCl at -20 degrees C are less suitable for use in feline ZBA. PMID- 17908297 TI - Primary DNA damage and genetic polymorphisms for CYP1A1, EPHX and GSTM1 in workers at a graphite electrode manufacturing plant. AB - BACKGROUND: The results of a cross-sectional study aimed to evaluate whether genetic polymorphisms (biomarkers of susceptibility) for CYP1A1, EPHX and GSTM1 genes that affect polycyclic aromatic hydrocarbons (PAH) activation and detoxification might influence the extent of primary DNA damage (biomarker of biologically effective dose) in PAH exposed workers are presented. PAH-exposure of the study populations was assessed by determining the concentration of 1 hydroxypyrene (1OHP) in urine samples (biomarker of exposure dose). METHODS: The exposed group consisted of workers (n = 109) at a graphite electrode manufacturing plant, occupationally exposed to PAH. Urinary 1OHP was measured by HPLC. Primary DNA damage was evaluated by the alkaline comet assay in peripheral blood leukocytes. Genetic polymorphisms for CYP1A1, EPHX and GSTM1 were determined by PCR or PCR/RFLP analysis. RESULTS: 1OHP and primary DNA damage were significantly higher in electrode workers compared to reference subjects. Moreover, categorization of subjects as normal or outlier highlighted an increased genotoxic risk OR = 2.59 (CI95% 1.32-5.05) associated to exposure to PAH. Polymorphisms in EPHX exons 3 and 4 was associated to higher urinary concentrations of 1OHP, whereas none of the genotypes analyzed (CYP1A1, EPHX, and GSTM1) had any significant influence on primary DNA damage as evaluated by the comet assay. CONCLUSION: The outcomes of the present study show that molecular epidemiology approaches (i.e. cross-sectional studies of genotoxicity biomarkers) can play a role in identifying common genetic risk factors, also attempting to associate the effects with measured exposure data. Moreover, categorization of subjects as normal or outlier allowed the evaluation of the association between occupational exposure to PAH and DNA damage highlighting an increased genotoxic risk. PMID- 17908300 TI - X-ray structures of two proteins belonging to Pfam DUF178 revealed unexpected structural similarity to the DUF191 Pfam family. AB - BACKGROUND: Pfam is a comprehensive collection of protein domains and families, with a range of well-established information including genome annotation. Pfam has two large series of functionally uncharacterized families, known as Domains of Unknown Function (DUFs) and Uncharacterized Protein Families (UPFs). RESULTS: Crystal structures of two proteins from Deinococcus radiodurans and Streptomyces coelicolor belonging to Pfam protein family DUF178 (ID: PF02621) have been determined using Selenium-Single-wavelength Anomalous Dispersion (Se-SAD). Based on the structure, we have identified the putative function for this family of protein. CONCLUSION: Unexpectedly, we found that DUF178 Pfam is remarkably similar to Pfam family DUF191 suggesting that the sequence-based classification alone may not be sufficient to classify proteins into Pfam families. PMID- 17908299 TI - HIV risk behaviors among female IDUs in developing and transitional countries. AB - BACKGROUND: A number of studies suggest females may be more likely to engage in injection and sex risk behavior than males. Most data on gender differences come from industrialized countries, so data are needed in developing countries to determine how well gender differences generalize to these understudied regions. METHODS: Between 1999 and 2003, 2512 male and 672 female current injection drug users (IDUs) were surveyed in ten sites in developing countries around the world (Nairobi, Beijing, Hanoi, Kharkiv, Minsk, St. Petersburg, Bogota, Gran Rosario, Rio, and Santos). The survey included a variety of questions about demographics, injecting practices and sexual behavior. RESULTS: Females were more likely to engage in risk behaviors in the context of a sexual relationship with a primary partner while males were more likely to engage in risk behaviors in the context of close friendships and casual sexual relationships. After controlling for injection frequency, and years injecting, these gender differences were fairly consistent across sites. CONCLUSION: Gender differences in risk depend on the relational contexts in which risk behaviors occur. The fact that female and male risk behavior often occurs in different relational contexts suggests that different kinds of prevention interventions which are sensitive to these contexts may be necessary. PMID- 17908301 TI - The prevalence and significance of substance use disorders in bipolar type I and II disorder. AB - The aim of this paper is to provide a systematic review of the literature examining the epidemiology, outcome, and treatment of patients with bipolar disorder and co-occurring substance use disorders (SUDs). Articles for this review were initially selected via a comprehensive Medline search and further studies were obtained from the references in these articles. Given the lack of research in this field, all relevant studies except case reports were included.Prior epidemiological research has consistently shown that substance use disorders (SUDs) are extremely common in bipolar I and II disorders. The lifetime prevalence of SUDs is at least 40% in bipolar I patients. Alcohol and cannabis are the substances most often abused, followed by cocaine and then opioids. Research has consistently shown that co-occurring SUDs are correlated with negative effects on illness outcome including more frequent and prolonged affective episodes, decreased compliance with treatment, a lower quality of life, and increased suicidal behavior. Recent research on the causal relationship between the two disorders suggests that a subgroup of bipolar patients may develop a relatively milder form of affective illness that is expressed only after extended exposure to alcohol abuse.There has been very little treatment research specifically targeting this population. Three open label medication trials provide limited evidence that quetiapine, aripiprazole, and lamotrigine may be effective in treating affective and substance use symptoms in bipolar patients with cocaine dependence and that aripiprazole may also be helpful in patients with alcohol use disorders. The two placebo controlled trials to date suggest that valproate given as an adjunct to lithium in bipolar patients with co occurring alcohol dependence improves both mood and alcohol use symptoms and that lithium treatment in bipolar adolescents improves mood and SUD symptoms.Given the high rate of SUD co-occurrence, more research investigating treatments in this population is needed. Specifically, double blind placebo controlled trials are needed to establish the effectiveness of medications found to be efficacious in open label treatments. New research also needs to be conducted on medications found to treat either bipolar disorder or a SUD in isolation. In addition, it may be advisable to consider including patients with prior SUDs in clinical trials for new medications in bipolar disorder. PMID- 17908302 TI - Paradoxical antiproliferative effect by a murine mammary tumor-derived epithelial cell line. AB - BACKGROUND: Despite significant advancement in breast cancer therapy, there is a great need for a better understanding of the mechanisms involved in breast carcinogenesis and progression, as well as of the role of epigenetic contributions from stromal cells in mammary tumorigenesis. In this study, we isolated and characterized murine mammary tumor-derived epithelial and myofibroblast cell lines, and investigated the in vitro and in vivo effect of cellular soluble factors produced by the epithelial cell line on tumor cells. METHODS: Morphology, immunophenotype, cytogenetics, invasiveness, and tumorigenicity of epithelial (LM-234ep) and myofibroblast (LM-234mf) cell lines isolated from two murine mammary adenocarcinomas with common ancestor were studied. The in vitro effects of LM-234ep conditioned medium on proliferation, cell cycle distribution, and expression of cell cycle proteins, were investigated in LM-234mf cells, mouse melanoma cells (B16-F10), and human cervical adenocarcinoma cells (HeLa). The in vivo anti-tumor activity of LM-234ep conditioned media was evaluated in subcutaneous tumors formed in nude mice by B16 F10 and HeLa cells. RESULTS: LM-234ep cells were found to be cytokeratin positive and hipertriploid, whereas LM-234mf cells were alpha-smooth muscle actin positive and hypohexaploid. Chromosome aberrations were found in both cases. Only LM-234mf revealed to be invasive in vitro and to secrete active MMP-2, though neither of the cell types were able to produce progressing tumors. LM-234ep-derived factors were able to inhibit the in vitro growth of LM-234mf, B16-F10, and HeLa cells, inducing cell cycle arrest in G0/G1 phase. The administration of LM-234ep conditioned medium inhibited the growth of B16-F10 and HeLa tumors in nude mice. CONCLUSION: Our data suggest the existence of epithelial cell variants with tumor suppressive properties within mammary tumors. To our knowledge, this is the first report showing antiproliferative and antineoplastic activities induced by tumor derived epithelial cells. PMID- 17908303 TI - Optimising the analysis of transcript data using high density oligonucleotide arrays and genomic DNA-based probe selection. AB - BACKGROUND: Affymetrix GeneChip arrays are widely used for transcriptomic studies in a diverse range of species. Each gene is represented on a GeneChip array by a probe-set, consisting of up to 16 probe-pairs. Signal intensities across probe pairs within a probe-set vary in part due to different physical hybridisation characteristics of individual probes with their target labelled transcripts. We have previously developed a technique to study the transcriptomes of heterologous species based on hybridising genomic DNA (gDNA) to a GeneChip array designed for a different species, and subsequently using only those probes with good homology. RESULTS: Here we have investigated the effects of hybridising homologous species gDNA to study the transcriptomes of species for which the arrays have been designed. Genomic DNA from Arabidopsis thaliana and rice (Oryza sativa) were hybridised to the Affymetrix Arabidopsis ATH1 and Rice Genome GeneChip arrays respectively. Probe selection based on gDNA hybridisation intensity increased the number of genes identified as significantly differentially expressed in two published studies of Arabidopsis development, and optimised the analysis of technical replicates obtained from pooled samples of RNA from rice. CONCLUSION: This mixed physical and bioinformatics approach can be used to optimise estimates of gene expression when using GeneChip arrays. PMID- 17908304 TI - DNA copy number profiles of gastric cancer precursor lesions. AB - BACKGROUND: Chromosomal instability (CIN) is the most prevalent type of genomic instability in gastric tumours, but its role in malignant transformation of the gastric mucosa is still obscure. In the present study, we set out to study whether two morphologically distinct categories of gastric cancer precursor lesions, i.e. intestinal-type and pyloric gland adenomas, would carry different patterns of DNA copy number changes, possibly reflecting distinct genetic pathways of gastric carcinogenesis in these two adenoma types. RESULTS: Using a 5K BAC array CGH platform, we showed that the most common aberrations shared by the 11 intestinal-type and 10 pyloric gland adenomas were gains of chromosomes 9 (29%), 11q (29%) and 20 (33%), and losses of chromosomes 13q (48%), 6(48%), 5(43%) and 10 (33%). The most frequent aberrations in intestinal-type gastric adenoma were gains on 11q, 9q and 8, and losses on chromosomes 5q, 6, 10 and 13, whereas in pyloric gland gastric adenomas these were gains on chromosome 20 and losses on 5q and 6. However, no significant differences were observed between the two adenoma types. CONCLUSION: The results suggest that gains on chromosomes 8, 9q, 11q and 20, and losses on chromosomes 5q, 6, 10 and 13, likely represent early events in gastric carcinogenesis. The phenotypical entities, intestinal type and pyloric gland adenomas, however, do not differ significantly (P = 0.8) at the level of DNA copy number changes. PMID- 17908305 TI - Forelimb-hindlimb developmental timing changes across tetrapod phylogeny. AB - BACKGROUND: Tetrapods exhibit great diversity in limb structures among species and also between forelimbs and hindlimbs within species, diversity which frequently correlates with locomotor modes and life history. We aim to examine the potential relation of changes in developmental timing (heterochrony) to the origin of limb morphological diversity in an explicit comparative and quantitative framework. In particular, we studied the relative time sequence of development of the forelimbs versus the hindlimbs in 138 embryos of 14 tetrapod species spanning a diverse taxonomic, ecomorphological and life-history breadth. Whole-mounts and histological sections were used to code the appearance of 10 developmental events comprising landmarks of development from the early bud stage to late chondrogenesis in the forelimb and the corresponding serial homologues in the hindlimb. RESULTS: An overall pattern of change across tetrapods can be discerned and appears to be relatively clade-specific. In the primitive condition, as seen in Chondrichthyes and Osteichthyes, the forelimb/pectoral fin develops earlier than the hindlimb/pelvic fin. This pattern is either retained or re-evolved in eulipotyphlan insectivores (= shrews, moles, hedgehogs, and solenodons) and taken to its extreme in marsupials. Although exceptions are known, the two anurans we examined reversed the pattern and displayed a significant advance in hindlimb development. All other species examined, including a bat with its greatly enlarged forelimbs modified as wings in the adult, showed near synchrony in the development of the fore and hindlimbs. CONCLUSION: Major heterochronic changes in early limb development and chondrogenesis were absent within major clades except Lissamphibia, and their presence across vertebrate phylogeny are not easily correlated with adaptive phenomena related to morphological differences in the adult fore- and hindlimbs. The apparently conservative nature of this trait means that changes in chondrogenetic patterns may serve as useful phylogenetic characters at higher taxonomic levels in tetrapods. Our results highlight the more important role generally played by allometric heterochrony in this instance to shape adult morphology. PMID- 17908306 TI - Recurrent adenylation domain replacement in the microcystin synthetase gene cluster. AB - BACKGROUND: Microcystins are small cyclic heptapeptide toxins produced by a range of distantly related cyanobacteria. Microcystins are synthesized on large NRPS PKS enzyme complexes. Many structural variants of microcystins are produced simultaneously. A recombination event between the first module of mcyB (mcyB1) and mcyC in the microcystin synthetase gene cluster is linked to the simultaneous production of microcystin variants in strains of the genus Microcystis. RESULTS: Here we undertook a phylogenetic study to investigate the order and timing of recombination between the mcyB1 and mcyC genes in a diverse selection of microcystin producing cyanobacteria. Our results provide support for complex evolutionary processes taking place at the mcyB1 and mcyC adenylation domains which recognize and activate the amino acids found at X and Z positions. We find evidence for recent recombination between mcyB1 and mcyC in strains of the genera Anabaena, Microcystis, and Hapalosiphon. We also find clear evidence for independent adenylation domain conversion of mcyB1 by unrelated peptide synthetase modules in strains of the genera Nostoc and Microcystis. The recombination events replace only the adenylation domain in each case and the condensation domains of mcyB1 and mcyC are not transferred together with the adenylation domain. Our findings demonstrate that the mcyB1 and mcyC adenylation domains are recombination hotspots in the microcystin synthetase gene cluster. CONCLUSION: Recombination is thought to be one of the main mechanisms driving the diversification of NRPSs. However, there is very little information on how recombination takes place in nature. This study demonstrates that functional peptide synthetases are created in nature through transfer of adenylation domains without the concomitant transfer of condensation domains. PMID- 17908307 TI - Diagnosis of drug-induced renal tubular toxicity using global gene expression profiles. AB - Toxicogenomics can measure the expression of thousands of genes to identify changes associated with drug induced toxicities. It is expected that toxicogenomics can be an alternative or complementary approach in preclinical drug safety evaluation to identify or predict drug induced toxicities. One of the major concerns in applying toxicogenomics to diagnose or predict drug induced organ toxicity, is how generalizable the statistical classification model is when derived from small datasets? Here we presented that a diagnosis of kidney proximal tubule toxicity, measured by pathology, can successfully be achieved even with a study design of limited number of training studies or samples. We selected a total of ten kidney toxicants, designed the in life study with multiple dose and multiple time points to cover samples at doses and time points with or without concurrent toxicity. We employed SVM (Support Vector Machine) as the classification algorithm for the toxicogenomic diagnosis of kidney proximal tubule toxicity. Instead of applying cross validation methods, we used an independent testing set by dividing the studies or samples into independent training and testing sets to evaluate the diagnostic performance. We achieved a Sn (sensitivity) = 88% and a Sp (specificity) = 91%. The diagnosis performance underscores the potential application of toxicogenomics in a preclinical lead optimization process of drugs entering into development. PMID- 17908308 TI - Characterization of oligopeptide patterns in large protein sets. AB - BACKGROUND: Recent sequencing projects and the growth of sequence data banks enable oligopeptide patterns to be characterized on a genome or kingdom level. Several studies have focused on kingdom or habitat classifications based on the abundance of short peptide patterns. There have also been efforts at local structural prediction based on short sequence motifs. Oligopeptide patterns undoubtedly carry valuable information content. Therefore, it is important to characterize these informational peptide patterns to shed light on possible new applications and the pitfalls implicit in neglecting bias in peptide patterns. RESULTS: We have studied four classes of pentapeptide patterns (designated POP, NEP, ORP and URP) in the kingdoms archaea, bacteria and eukaryotes. POP are highly abundant patterns statistically not expected to exist; NEP are patterns that do not exist but are statistically expected to; ORP are patterns unique to a kingdom; and URP are patterns excluded from a kingdom. We used two data sources: the de facto standard of protein knowledge Swiss-Prot, and a set of 386 completely sequenced genomes. For each class of peptides we looked at the 100 most extreme and found both known and unknown sequence features. Most of the known sequence motifs can be explained on the basis of the protein families from which they originate. CONCLUSION: We find an inherent bias of certain oligopeptide patterns in naturally occurring proteins that cannot be explained solely on the basis of residue distribution in single proteins, kingdoms or databases. We see three predominant categories of patterns: (i) patterns widespread in a kingdom such as those originating from respiratory chain associated proteins and translation machinery; (ii) proteins with structurally and/or functionally favored patterns, which have not yet been ascribed this role; (iii) multicopy species-specific retrotransposons, only found in the genome set. These categories will affect the accuracy of sequence pattern algorithms that rely mainly on amino acid residue usage. Methods presented in this paper may be used to discover targets for antibiotics, as we identify numerous examples of kingdom-specific antigens among our peptide classes. The methods may also be useful for detecting coding regions of genes. PMID- 17908309 TI - Changing population characteristics, effect-measure modification, and cancer risk factor identification. AB - Epidemiologic studies have identified a number of lifestyle factors, e.g. diet, obesity, and use of certain medications, which affect risk of colon cancer. However, the magnitude and significance of risk factor-disease associations differ among studies. We propose that population trends of changing prevalence of risk factors explains some of the variability between studies when factors that change prevalence also modify the effect of other risk factors. We used data collected from population-based control who were selected as study participants for two time periods, 1991-1994 and 1997-2000, along with data from the literature, to examine changes in the population prevalence of aspirin and non steroidal anti-inflammatory medication (NSAID) use, obesity, and hormone replacement therapy (HRT) over time. Data from a population-based colon cancer case-control study were used to estimate effect-measurement modification among these factors. Sizeable changes in aspirin use, HRT use, and the proportion of the population that is obese were observed between the 1980s and 2000. Use of NSAIDs interacted with BMI and HRT; HRT use interacted with body mass index (BMI). We estimate that as the prevalence of NSAIDs use changed from 10% to almost 50%, the colon cancer relative risk associated with BMI >30 would change from 1.3 to 1.9 because of the modifying effect of NSAIDs. Similarly, the relative risk estimated for BMI would increase as the prevalence of use of HRT among post-menopausal women increased. In conclusion, as population characteristics change over time, these changes may have an influence on relative risk estimates for colon cancer for other exposures because of effect-measure modification. The impact of population changes on comparability between epidemiologic studies can be kept to a minimum if investigators assess exposure disease associations within strata of other exposures, and present results in a manner that allows comparisons across studies. Effect-measure modification is an important component of data analysis that should be evaluated to obtain a complete understanding of disease etiology. PMID- 17908310 TI - PCR-based karyotyping of Anopheles gambiae inversion 2Rj identifies the BAMAKO chromosomal form. AB - BACKGROUND: The malaria vector Anopheles gambiae is polymorphic for chromosomal inversions on the right arm of chromosome 2 that segregate nonrandomly between assortatively mating populations in West Africa. One such inversion, 2Rj, is associated with the BAMAKO chromosomal form endemic to southern Mali and northern Guinea Conakry near the Niger River. Although it exploits a unique ecology and both molecular and chromosomal data suggest reduced gene flow between BAMAKO and other A. gambiae populations, no molecular markers exist to identify this form. METHODS: To facilitate study of the BAMAKO form, a PCR assay for molecular karyotyping of 2Rj was developed based on sequences at the breakpoint junctions. The assay was extensively validated using more than 700 field specimens whose karyotypes were determined in parallel by cytogenetic and molecular methods. As inversion 2Rj also occurs in SAVANNA populations outside the geographic range of BAMAKO, samples were tested from Senegal, Cameroon and western Guinea Conakry as well as from Mali. RESULTS: In southern Mali, where 2Rj polymorphism in SAVANNA populations was very low and most of the 2Rj homozygotes were found in BAMAKO karyotypes, the molecular and cytogenetic methods were almost perfectly congruent. Elsewhere agreement between the methods was much poorer, as the molecular assay frequently misclassified 2Rj heterozygotes as 2R+j standard homozygotes. CONCLUSION: Molecular karyotyping of 2Rj is robust and accurate on 2R+j standard and 2Rj inverted homozygotes. Therefore, the proposed approach overcomes the lack of a rapid tool for identifying the BAMAKO form across developmental stages and sexes, and opens new perspectives for the study of BAMAKO ecology and behaviour. On the other hand, the method should not be applied for molecular karyotyping of j-carriers within the SAVANNA chromosomal form. PMID- 17908311 TI - Dual antiplatelet therapy and drug eluting stents: a marriage of convenience. PMID- 17908313 TI - Emotional support, education and self-rated health in 22 European countries. AB - BACKGROUND: The analyses focus on three aims: (1) to explore the associations between education and emotional support in 22 European countries, (2) to explore the associations between emotional support and self-rated health in the European countries, and (3) to analyse whether the association between education and self rated health can be partly explained by emotional support. METHODS: The study uses data from the European Social Survey 2003. Probability sampling from all private residents aged 15 years and older was applied in all countries. The European Social Survey includes 42,359 cases. Persons under age 25 were excluded to minimise the number of respondents whose education was not complete. Education was coded according to the International Standard Classification of Education. Perceived emotional support was assessed by the availability of a confidant with whom one can discuss intimate and personal matters with. Self-rated health was used as health indicator. RESULTS: Results of multiple logistic regression analyses show that emotional support is positively associated with education among women and men in most European countries. However, the magnitude of the association varies according to country and gender. Emotional support is positively associated with self-rated health. Again, gender and country differences in the association were observed. Emotional support explains little of the educational differences in self-rated health among women and men in most European countries. CONCLUSION: Results indicate that it is important to consider socio-economic factors like education and country-specific contexts in studies on health effects of emotional support. PMID- 17908312 TI - Human immunodeficiency virus type 1 Nef protein modulates the lipid composition of virions and host cell membrane microdomains. AB - BACKGROUND: The Nef protein of Human Immunodeficiency Viruses optimizes viral spread in the infected host by manipulating cellular transport and signal transduction machineries. Nef also boosts the infectivity of HIV particles by an unknown mechanism. Recent studies suggested a correlation between the association of Nef with lipid raft microdomains and its positive effects on virion infectivity. Furthermore, the lipidome analysis of HIV-1 particles revealed a marked enrichment of classical raft lipids and thus identified HIV-1 virions as an example for naturally occurring membrane microdomains. Since Nef modulates the protein composition and function of membrane microdomains we tested here if Nef also has the propensity to alter microdomain lipid composition. RESULTS: Quantitative mass spectrometric lipidome analysis of highly purified HIV-1 particles revealed that the presence of Nef during virus production from T lymphocytes enforced their raft character via a significant reduction of polyunsaturated phosphatidylcholine species and a specific enrichment of sphingomyelin. In contrast, Nef did not significantly affect virion levels of phosphoglycerolipids or cholesterol. The observed alterations in virion lipid composition were insufficient to mediate Nef's effect on particle infectivity and Nef augmented virion infectivity independently of whether virus entry was targeted to or excluded from membrane microdomains. However, altered lipid compositions similar to those observed in virions were also detected in detergent resistant membrane preparations of virus producing cells. CONCLUSION: Nef alters not only the proteome but also the lipid composition of host cell microdomains. This novel activity represents a previously unrecognized mechanism by which Nef could manipulate HIV-1 target cells to facilitate virus propagation in vivo. PMID- 17908314 TI - Homozygosity by descent mapping of blood pressure in the Old Order Amish: evidence for sex specific genetic architecture. AB - BACKGROUND: High blood pressure is a well established risk factor for morbidity and mortality acting through heart disease, stroke and cardiovascular disease. Genome wide scans have linked regions of nearly every human chromosome to blood pressure related traits. We have capitalized on beneficial qualities of the Old Order Amish of Lancaster, PA, a closed founder population with a relatively small number of founders, to perform a genome wide homozygosity by descent mapping scan. Each individual in the study has a non zero probability of consanguinity. Systolic and diastolic blood pressures are shown to have appreciable dominance variance components. RESULTS: Areas of two chromosomes were identified as suggestive of linkage to SBP and 5 areas to DBP in either the overall or sex specific analyses. The strongest evidence for linkage in the overall sample was to Chromosome 18q12 (LOD = 2.6 DBP). Sex specific analyses identified a linkage on Chromosome 4p12-14 (LOD in men only = 3.4 SBP). At Chromosome 2q32-33, an area where we previously reported significant evidence for linkage to DBP using a conventional identity by descent approach, the LOD was 1.4; however an appreciable sex effect was observed with men accounting for most of the linkage (LOD in men only = 2.6). CONCLUSION: These results add evidence to a sex specific genetic architecture to blood pressure related traits, particularly in regions of linkage on chromosome 2, 4 and 18. PMID- 17908315 TI - The cost-effectiveness of a school-based overweight program. AB - BACKGROUND: This study assesses the net benefit and the cost-effectiveness of the Coordinated Approach to Child Health (CATCH) intervention program, using parameter estimates from the El Paso trial. There were two standard economic measures used. First, from a societal perspective on costs, cost-effectiveness ratios (CER) were estimated, revealing the intervention costs per quality adjusted life years (QALYs) saved. QALY weights were estimated using National Health Interview Survey (NHIS) data. Second, the net benefit (NB) of CATCH was estimated, which compared the present value of averted future costs with the cost of the CATCH intervention. Using National Health and Nutrition Examination Survey I (NHANES) and NHANES follow-up data, we predicted the number of adult obesity cases avoided for ages 40-64 with a lifetime obesity progression model. RESULTS: The results show that CATCH is cost-effective and net beneficial. The CER was US$900 (US$903 using Hispanic parameters) and the NB was US$68,125 (US$43,239 using Hispanic parameters), all in 2004 dollars. This is much lower than the benchmark for CER of US$30,000 and higher than the NB of US$0. Both were robust to sensitivity analyses. CONCLUSION: Childhood school-based programs such as CATCH are beneficial investments. Both NB and CER declined when Hispanic parameters were included, primarily due to the lower wages earned by Hispanics. However, both NB and CER for Hispanics were well within standard cost effectiveness and net benefit thresholds. PMID- 17908316 TI - Limited genetic diversity in Salmonella enterica serovar Enteritidis PT13. AB - BACKGROUND: Salmonella enterica serovar Enteritidis has emerged as a significant foodborne pathogen throughout the world and is commonly characterized by phage typing. In Canada phage types (PT) 4, 8 and 13 predominate and in 2005 a large foodborne PT13 outbreak occurred in the province of Ontario. The ability to link strains during this outbreak was difficult due to the apparent clonality of PT13 isolates in Canada, as there was a single dominant pulsed-field gel electrophoresis (PFGE) profile amongst epidemiologically linked human and food isolates as well as concurrent sporadic strains. The aim of this study was to perform comparative genomic hybridization (CGH), DNA sequence-based typing (SBT) genomic analyses, plasmid analyses, and automated repetitive sequence-based PCR (rep-PCR) to identify epidemiologically significant traits capable of subtyping S. Enteritidis PT13. RESULTS: CGH using an oligonucleotide array based upon chromosomal coding sequences of S. enterica serovar Typhimurium strain LT2 and the Salmonella genomic island 1 successfully determined major genetic differences between S. Typhimurium and S. Enteritidis PT13, but no significant strain-to strain differences were observed between S. Enteritidis PT13 isolates. Individual loci (safA and fliC) that were identified as potentially divergent in the CGH data set were sequenced in a panel of S. Enteritidis strains, and no differences were detected between the PT13 strains. Additional sequence-based typing was performed at the fimA, mdh, manB, cyaA, citT, caiC, dmsA, ratA and STM0660 loci. Similarly, no diversity was observed amongst PT13 strains. Variation in plasmid content between PT13 strains was observed, but macrorestriction with BglII did not identify further differences. Automated rep-PCR patterns were variable between serovars, but S. Enteritidis PT13 strains could not be differentiated. CONCLUSION: None of the methods identified any significant variation between PT13 strains. Greater than 11,300 base pairs of sequence for each of seven S. Enteritidis PT13 strains were analyzed without detecting a single polymorphic site, although diversity between different phage types of S. Enteritidis was observed. These data suggest that Canadian S. Enteritidis PT13 strains are highly related genetically. PMID- 17908317 TI - Effect of cough technique and cryogen gas on temperatures achieved during simulated cryotherapy. AB - BACKGROUND: Cryotherapy is often used to treat cervical precancer in developing countries. There are different methods and cryogen gases used for cryotherapy, including the freeze-flush-freeze (cough) technique employed to minimize gas blockage. However, there is limited information to compare their effectiveness. METHODS: Using a tissue model, we compared temperature-time curves for four cryotherapy methods: uninterrupted freezing with nitrous oxide (N2O) and carbon dioxide (CO2), and two methods using a standard and extended version of the cough technique with CO2. RESULTS: Uninterrupted freezing with both N2O and CO2 produced tissue temperatures less than -20 degrees C (-40 degrees C and -30 degrees C respectively). CO2 cryotherapy procedures using the two cough techniques produced temperatures greater than -20 degrees C in the model tissue. CONCLUSION: CO2 cryotherapy using the cough technique may not achieve sufficiently low temperatures to produce the desired therapeutic effect. Other alternatives to the prevention of gas blockage should be developed. PMID- 17908318 TI - RNAspa: a shortest path approach for comparative prediction of the secondary structure of ncRNA molecules. AB - BACKGROUND: In recent years, RNA molecules that are not translated into proteins (ncRNAs) have drawn a great deal of attention, as they were shown to be involved in many cellular functions. One of the most important computational problems regarding ncRNA is to predict the secondary structure of a molecule from its sequence. In particular, we attempted to predict the secondary structure for a set of unaligned ncRNA molecules that are taken from the same family, and thus presumably have a similar structure. RESULTS: We developed the RNAspa program, which comparatively predicts the secondary structure for a set of ncRNA molecules in linear time in the number of molecules. We observed that in a list of several hundred suboptimal minimal free energy (MFE) predictions, as provided by the RNAsubopt program of the Vienna package, it is likely that at least one suggested structure would be similar to the true, correct one. The suboptimal solutions of each molecule are represented as a layer of vertices in a graph. The shortest path in this graph is the basis for structural predictions for the molecule. We also show that RNA secondary structures can be compared very rapidly by a simple string Edit-Distance algorithm with a minimal loss of accuracy. We show that this approach allows us to more deeply explore the suboptimal structure space. CONCLUSION: The algorithm was tested on three datasets which include several ncRNA families taken from the Rfam database. These datasets allowed for comparison of the algorithm with other methods. In these tests, RNAspa performed better than four other programs. PMID- 17908319 TI - Abundance and functional diversity of riboswitches in microbial communities. AB - BACKGROUND: Several recently completed large-scale enviromental sequencing projects produced a large amount of genetic information about microbial communities ('metagenomes') which is not biased towards cultured organisms. It is a good source for estimation of the abundance of genes and regulatory structures in both known and unknown members of microbial communities. In this study we consider the distribution of RNA regulatory structures, riboswitches, in the Sargasso Sea, Minnesota Soil and Whale Falls metagenomes. RESULTS: Over three hundred riboswitches were found in about 2 Gbp metagenome DNA sequences. The abundabce of riboswitches in metagenomes was highest for the TPP, B12 and GCVT riboswitches; the S-box, RFN, YKKC/YXKD, YYBP/YKOY regulatory elements showed lower but significant abundance, while the LYS, G-box, GLMS and YKOK riboswitches were rare. Regions downstream of identified riboswitches were scanned for open reading frames. Comparative analysis of identified ORFs revealed new riboswitch regulated functions for several classes of riboswitches. In particular, we have observed phosphoserine aminotransferase serC (COG1932) and malate synthase glcB (COG2225) to be regulated by the glycine (GCVT) riboswitch; fatty acid desaturase ole1 (COG1398), by the cobalamin (B12) riboswitch; 5-methylthioribose-1-phosphate isomerase ykrS (COG0182), by the SAM-riboswitch. We also identified conserved riboswitches upstream of genes of unknown function: thiamine (TPP), cobalamine (B12), and glycine (GCVT, upstream of genes from COG4198). CONCLUSION: This study demonstrates applicability of bioinformatics to the analysis of RNA regulatory structures in metagenomes. PMID- 17908321 TI - A pharmacodynamic analysis of resistance trends in pathogens from patients with infection in intensive care units in the United States between 1993 and 2004. AB - BACKGROUND: Increasing nosocomial pathogen resistance to available antimicrobial agents is of growing concern. While higher MICs can diminish antimicrobial effectiveness, dose adjustments often mitigate this effect. This study's objective was to ascertain whether MICs among major pathogens in the ICU to several commonly used agents have increased enough to significantly impact their ability to achieve bactericidal effect. METHODS: Cefepime, ceftriaxone, imipenem and piperacillin-tazobactam MICs were determined with 74,394 Gram-negative bacilli obtained from ICU patients with various infections in the US between 1993 and 2004. Results were grouped into four 3-year periods. The predicted cumulative fraction of response (CFR) was estimated based on patient-derived pharmacokinetic values and Monte Carlo simulation. Trends in CFR over the four study periods were assessed using the Cochran-Armitage test. The primary analysis included all organisms combined; Pseudomonas aeruginosa and Acinetobacter species were also evaluated individually. RESULTS: In the primary analysis, imipenem 500 mg q6h showed CFRs from 87% to 90% across all four study periods, with a trend toward slightly improved bactericidal target attainment (p < 0.01). CFRs for cefepime 2 g q12h and piperacillin-tazobactam 4.5 g q6h both declined by 2% (p < 0.01 and p < 0.05, respectively), reflecting upward shifts in the underlying MIC distributions. Ceftriaxone had <52% CFR for all regimens in all periods, with no significant trend. Against P. aeruginosa, significant declines in CFR were seen for (range, p-value): imipenem 1 g q8h (82%-79%, p < 0.01), cefepime 1 g q12h (70%-67%, p < 0.01), cefepime 2 g q12h (84%-82%, p < 0.05), piperacillin tazobactam 3.375 g q6h (76%-73%, p < 0.01), piperacillin-tazobactam 4.5 g q8h (71%-68%, p < 0.01), and piperacillin-tazobactam 4.5 g q6h (80%-77%, p < .01). Against Acinetobacter spp., all regimens of imipenem, cefepime and piperacillin tazobactam showed significant declines in CFR over time (p < 0.01). CONCLUSION: Our observations suggest that as a result of increasing antimicrobial resistance among ICU pathogens in the US, drug effectiveness, assessed as a function of individual agents' ability to attain pharmacodynamic targets, has declined, especially with P. aeruginosa and Acinetobacter spp. Cefepime 2 g q8h and imipenem were the most potent agents against these species, respectively. More aggressive dosing of all of the agents characterized could preserve their clinical utility, but this must be balanced with safety and tolerability issues by the physician. PMID- 17908320 TI - RBM6-RBM5 transcription-induced chimeras are differentially expressed in tumours. AB - Transcription-induced chimerism, a mechanism involving the transcription and intergenic splicing of two consecutive genes, has recently been estimated to account for approximately 5% of the human transcriptome. Despite this prevalence, the regulation and function of these fused transcripts remains largely uncharacterised. RESULTS: We identified three novel transcription-induced chimeras resulting from the intergenic splicing of a single RNA transcript incorporating the two neighbouring 3p21.3 tumour suppressor locus genes, RBM6 and RBM5, which encode the RNA Binding Motif protein 6 and RNA Binding Motif protein 5, respectively. Each of the three novel chimeric transcripts lacked exons 3, 6, 20 and 21 of RBM6 and exon 1 of RBM5. Differences between the transcripts were associated with the presence or absence of exon 4, exon 5 and a 17 nucleotide (nt) sequence from intron 10 of RBM6. All three chimeric transcripts incorporated the canonical splice sites from both genes (excluding the 17 nt intron 10 insertion). Differential expression was observed in tumour tissue compared to non tumour tissue, and amongst tumour types. In breast tumour tissue, chimeric expression was associated with elevated levels of RBM6 and RBM5 mRNA, and increased tumour size. No protein expression was detected by in vitro transcription/translation. CONCLUSION: These results suggest that RBM6 mRNA experiences altered co-transcriptional gene regulation in certain cancers. The results also suggest that RBM6-RBM5 transcription-induced chimerism might be a process that is linked to the tumour-associated increased transcriptional activity of the RBM6 gene. It appears that none of the transcription-induced chimeras generates a protein product; however, the novel alternative splicing, which affects putative functional domains within exons 3, 6 and 11 of RBM6, does suggest that the generation of these chimeric transcripts has functional relevance. Finally, the association of chimeric expression with breast tumour size suggests that RBM6-RBM5 chimeric expression may be a potential tumour differentiation marker. PMID- 17908322 TI - Cost-effectiveness of the SEN-concept: Specialized Emergency Nurses (SEN) treating ankle/foot injuries. AB - BACKGROUND: Emergency Departments (EDs) are confronted with progressive overcrowding. As a consequence, the workload for ED physicians increases and waiting times go up with the risk of unnecessary complications and patient dissatisfaction. To cope with these problems, Specialized Emergency Nurses (SENs), regular ED-nurses receiving a short, injury-specific course, were trained to assess and treat minor injuries according to a specific protocol. METHODS: An economic evaluation was conducted alongside a randomized controlled trial comparing House Officers (HOs) and SENs in their assessment of ankle and foot injuries. Cost prices were established for all parts of healthcare utilization involved. Total costs of health care utilization were computed per patient in both groups. Cost-effectiveness was investigated by comparing the difference in total cost between groups with the difference in sensitivity and specificity between groups in diagnosing fractures and severe sprains. Finally, cost effectiveness ratios were calculated and presented on a cost-effectiveness plane. RESULTS: No significant differences were seen between treatment groups for any of the health care resources assessed. However, the waiting times for both first assessment by a treatment officer and time spent waiting between hearing the diagnosis and final treatment were significantly longer in the HO group. There was no statistically significant difference in costs between groups. The total costs were euro 186 (SD euro 623) for patients in the SEN group and euro 153 (SD euro 529) for patients in the HO group. The difference in total costs was euro 33 (95% CI: - euro 84 to euro 155). The incremental cost-effectiveness ratio was euro 27 for a reduction of one missed diagnosis and euro 18 for a reduction of one false negative. CONCLUSION: Considering the benefits of the SEN-concept in terms of decreased workload for the ED physicians, increased patient satisfaction and decreased waiting times, SENs appear to be a useful solution to the problem of ED crowding. PMID- 17908323 TI - Conservative management of breast cancer in the elderly in a developing country. AB - BACKGROUND: The cost effective treatment of cancer in developing countries remains challenging. In the elderly with possible limited life expectancy, the health expenditure associated with standard treatment regimes should be carefully considered. We present the results of conservative management of breast cancer in the aged in a resource-limited environment. METHODS: Patients aged 70 or older with early breast cancer were treated with tumour excision or simple mastectomy and adjuvant tamoxifen. The records of patients presenting to the Breast Unit between January 1990 and December 2004 were retrieved and demographic, clinical, pathological and oncological data were reviewed. Survival statistics were calculated using the life table method. RESULTS: A total of 483 patients above 70 years of age were identified. One hundred and eighty eight patients were managed according to the conservative protocol. Forty-one had a simple mastectomy and 147 tumour excision. Their mean age was 77.3 years. The mean follow-up is 62 months. Thirty-one patients (16.4%) were not compliant with tamoxifen use. TNM staging was 0 in 4 patients, I in 42 patients, II in 116 patients and III in 26 patients. There was no 30-day mortality. The cumulative incidence of local recurrence was 3.3% at 5 and 10 years. The cumulative incidence of regional recurrence was 3.3% at 5 years and 4.5% at 10 years. The cumulative incidence of distant recurrence was 6.2% at 5 years and 12.2% at 10 years. The cumulative overall, disease specific and disease free survival at 10 years was 59%, 88% and 81% respectively. CONCLUSION: Limited surgery and tamoxifen provide excellent control of breast cancer in the elderly in a resource restricted environment. Radiotherapy and axillary dissection and can be safely omitted thereby reducing health care resource utilization. PMID- 17908324 TI - Comparison of different commercial kits for HER2 testing in breast cancer: looking for the accurate cutoff for amplification. AB - INTRODUCTION: Accurate determination of human epidermal growth factor receptor 2 (HER2) status is essential for optimal patient management with trastuzumab (Herceptin). However, standard guidelines do not specify a particular commercial kit, antibody or probe for testing, and discrepancies arise from variability between kits. The aim of this study was to compare the accuracy of four commercially available fluorescence/chromogenic in situ hybridisation (FISH/CISH) kits and validate one for the resolution of borderline immunohistochemistry (IHC) cases. The interpretation pitfalls, optimal threshold values, assay duration and complexity of each kit were also considered. METHODS: The Food and Drug Administration (FDA)-approved dual-probe FISH assay PathVysion was chosen as the 'gold standard' against which pharmDx (dual-probe) and INFORM (mono-probe) FDA approved FISH kits and the SPoT-Light CISH kit were compared. Tumours were also evaluated by IHC with the FDA-approved HercepTest kit and a validated in-house IHC protocol. Fifty-five patients with invasive breast carcinoma were selected as a representative proportion of HER2 IHC 2+ cases. RESULTS: HER2 amplification was observed in 31% of tumours by PathVysion compared with 33% with pharmDx. The number of amplified tumours detected by INFORM and CISH varied with the threshold applied. Agreement was excellent between PathVysion and pharmDx (100%), good with SPoT-Light (89%; cutoff at least five signals per nucleus) and moderate with INFORM (76%; cutoff more than four signals per nucleus). Agreement with INFORM improved to 98% with a cutoff of at least six signals per nucleus. CONCLUSION: With an appropriate cutoff, the INFORM kit was comparable to dual-probe FISH kits for evaluating HER2 status. We validate and recommend CISH as an appropriate assay for HER2 scoring that is easy to interpret and requires equipment readily found in, or that can be adapted to, all pathology laboratories. For borderline IHC cases, dual-probe FISH analysis remains the most useful protocol to apply. PMID- 17908325 TI - Deleted in Malignant Brain Tumors 1 (DMBT1) is present in hyaline membranes and modulates surface tension of surfactant. AB - BACKGROUND: Deleted in Malignant Brain Tumors 1 (DMBT1) is a secreted scavenger receptor cysteine-rich protein that binds various bacteria and is thought to participate in innate pulmonary host defense. We hypothesized that pulmonary DMBT1 could contribute to respiratory distress syndrome in neonates by modulating surfactant function. METHODS: DMBT1 expression was studied by immunohistochemistry and mRNA in situ hybridization in post-mortem lungs of preterm and full-term neonates with pulmonary hyaline membranes. The effect of human recombinant DMBT1 on the function of bovine and porcine surfactant was measured by a capillary surfactometer. DMBT1-levels in tracheal aspirates of ventilated preterm and term infants were determined by ELISA. RESULTS: Pulmonary DMBT1 was localized in hyaline membranes during respiratory distress syndrome. In vitro addition of human recombinant DMBT1 to the surfactants increased surface tension in a dose-dependent manner. The DMBT1-mediated effect was reverted by the addition of calcium depending on the surfactant preparation. CONCLUSION: Our data showed pulmonary DMBT1 expression in hyaline membranes during respiratory distress syndrome and demonstrated that DMBT1 increases lung surface tension in vitro. This raises the possibility that DMBT1 could antagonize surfactant supplementation in respiratory distress syndrome and could represent a candidate target molecule for therapeutic intervention in neonatal lung disease. PMID- 17908326 TI - Metal ion-dependent, reversible, protein filament formation by designed beta-roll polypeptides. AB - BACKGROUND: A right-handed, calcium-dependent beta-roll structure found in secreted proteases and repeat-in-toxin proteins was used as a template for the design of minimal, soluble, monomeric polypeptides that would fold in the presence of Ca2+. Two polypeptides were synthesised to contain two and four metal binding sites, respectively, and exploit stacked tryptophan pairs to stabilise the fold and report on the conformational state of the polypeptide. RESULTS: Initial analysis of the two polypeptides in the presence of calcium suggested the polypeptides were disordered. The addition of lanthanum to these peptides caused aggregation. Upon further study by right angle light scattering and electron microscopy, the aggregates were identified as ordered protein filaments that required lanthanum to polymerize. These filaments could be disassembled by the addition of a chelating agent. A simple head-to-tail model is proposed for filament formation that explains the metal ion-dependency. The model is supported by the capping of one of the polypeptides with biotin, which disrupts filament formation and provides the ability to control the average length of the filaments. CONCLUSION: Metal ion-dependent, reversible protein filament formation is demonstrated for two designed polypeptides. The polypeptides form filaments that are approximately 3 nm in diameter and several hundred nm in length. They are not amyloid-like in nature as demonstrated by their behaviour in the presence of congo red and thioflavin T. A capping strategy allows for the control of filament length and for potential applications including the "decoration" of a protein filament with various functional moieties. PMID- 17908327 TI - Association between erythrocyte Na+K+-ATPase activity and some blood lipids in type 1 diabetic patients from Lagos, Nigeria. AB - BACKGROUND: Altered levels of erythrocyte Na+K+-ATPase, atherogenic and anti atherogenic lipid metabolites have been implicated in diabetic complications but their pattern of interactions remains poorly understood.This study evaluated this relationship in Nigerian patients with Type 1 diabetes mellitus. METHODS: A total of 34 consented Type 1 diabetic patients and age -matched 27 non-diabetic controls were enrolled. Fasting plasma levels of total cholesterol, triglycerides and HDL-cholesterol were determined spectrophotometrically and LDL-cholesterol estimated using Friedewald formula. Total protein content and Na+K+-ATPase activity were also determined spectrophotometrically from ghost erythrocyte membrane prepared by osmotic lysis. RESULTS: Results indicate significant (P < 0.05) reduction in Na+K+-ATPase activity in the Type 1 diabetic patients (0.38 +/ 0.08 vs. 0.59 +/- 0.07 microM Pi/mgprotein/h) compared to the control but with greater reduction in the diabetic subgroup with poor glycemic control (n = 20) and in whom cases of hypercholesterolemia (8.8%), hypertriglyceridemia (2.9%) and elevated LDL-cholesterol (5.9% each) were found. Correlation analyses further revealed significant (P < 0.05) inverse correlations [r = -(0.708-0.797] between all the atherogenic lipid metabolites measured and Na+K+-ATPase in this subgroup contrary to group with good glycemic control or non-diabetic subjects in which significant (P < 0.05) Na+K+-ATPase and HDL-C association were found (r = 0.427 - 0.489). The Na+K+-ATPase from the diabetic patients also exhibited increased sensitivity to digoxin and alterations in kinetic constants Vmax and Km determined by glycemic status of the patients. CONCLUSION: It can be concluded that poor glycemic control evokes greater reduction in erythrocyte Na+K+-ATPase activity and promote enzyme-blood atherogenic lipid relationships in Type 1 diabetic Nigerian patients. PMID- 17908328 TI - Direct visualization of a significant stenosis of the right coronary artery by transthoracic echocardiography. A case report. AB - Non-invasive imaging of coronary arteries by transthoracic echocardiography is an emerging diagnostic tool to study the left main (LM), left descending artery (LAD), circumflex (Cx) and right coronary artery (RCA). Impaired coronary circulation can be assessed by measuring coronary velocity flow reserve (CVFR) by transthoracic Doppler echocardiography. Coronary artery stenoses can be identified as localized colour aliasing and accelerated flow velocities. We report a case with an acute coronary syndrome (ACS) of a 46-year-old man. With non-invasive imaging of coronary arteries by transthoracic echocardiography (TTE), we identified a segment of the mid right coronary artery (RCA) suggestive of stenosis with localized colour aliasing and accelerated flow velocity. We found a high ratio between the stenotic peak velocity and the prestenotic peak velocity, and a pathologic coronary flow velocity reserve (CFVR) distal to the stenosis in the posterior interventricular descending branch (RDP). Subsequent coronary angiography demonstrated one vessel disease with a stenosis in segment 3 of RCA, which was successfully treated with percutaneos coronary intervention PCI. Two weeks following the PCI procedure he was readmitted to hospital with chest pain. A subacute stent thrombosis was questioned, and repeated echocardiography was preformed. The mid portion of RCA showed normal and laminar flow. The CVFR of RCA measured in the RDP showed normal vasodilatory response, confirming an open RCA without any flow limitation. A repeated coronary angiogram demonstrated only a mild in stent intimal hyperplasia. This case illustrates the value of transthoracic echocardiography as a tool both in the diagnosis and the follow-up of chest pain disorders and coronary flow problems. Transthoracic echocardiography allows both direct visualization of the various coronary segments and assessment of the CVFR. PMID- 17908330 TI - Agmatine protects retinal ganglion cells from hypoxia-induced apoptosis in transformed rat retinal ganglion cell line. AB - BACKGROUND: Agmatine is an endogenous polyamine formed by the decarboxylation of L-arginine. We investigated the protective effects of agmatine against hypoxia induced apoptosis of immortalized rat retinal ganglion cells (RGC-5). RGC-5 cells were cultured in a closed hypoxic chamber (5% O2) with or without agmatine. Cell viability was determined by lactate dehydrogenase (LDH) assay and apoptosis was examined by annexin V and caspase-3 assays. Expression and phosphorylation of mitogen-activated protein kinases (MAPKs; JNK, ERK p44/42, and p38) and nuclear factor-kappa B (NF-kappaB) were investigated by Western immunoblot analysis. The effects of agmatine were compared to those of brain-derived neurotrophic factor (BDNF), a well-known protective neurotrophin for retinal ganglion cells. RESULTS: After 48 hours of hypoxic culture, the LDH assay showed 52.3% cell loss, which was reduced to 25.6% and 30.1% when agmatine and BDNF were administered, respectively. This observed cell loss was due to apoptotic cell death, as established by annexin V and caspase-3 assays. Although total expression of MAPKs and NF-kappaB was not influenced by hypoxic injury, phosphorylation of these two proteins was increased. Agmatine reduced phosphorylation of JNK and NF-kappaB, while BDNF suppressed phosphorylation of ERK and p38. CONCLUSION: Our results show that agmatine has neuroprotective effects against hypoxia-induced retinal ganglion cell damage in RGC-5 cells and that its effects may act through the JNK and NF-kappaB signaling pathways. Our data suggest that agmatine may lead to a novel therapeutic strategy to reduce retinal ganglion cell injury related to hypoxia. PMID- 17908331 TI - Pelvic tenderness is not limited to the prostate in chronic prostatitis/chronic pelvic pain syndrome (CPPS) type IIIA and IIIB: comparison of men with and without CP/CPPS. AB - BACKGROUND: We wished to determine if there were differences in pelvic and non pelvic tenderness between men with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) Type III and men without pelvic pain. METHODS: We performed the Manual Tender Point Survey (MTPS) as described by the American College of Rheumatology on 62 men with CP/CPPS Type IIIA and IIIB and 98 men without pelvic pain. We also assessed tenderness of 10 external pelvic tender points (EPTP) and of 7 internal pelvic tender points (IPTP). All study participants completed the National Institutes of Health Chronic Prostatitis Symptom Inventory (NIH CPSI). RESULTS: We found that men with CPPS were significantly more tender in the MTPS, the EPTPS and the IPTPS. CPSI scores correlated with EPTP scale but not with IPTP scale or prostate tenderness. Prostatic tenderness was present in 75% of men with CPPS and in 50% of men without CPPS. Expressed prostatic fluid leukocytosis was not associated with prostatic tenderness. CONCLUSION: Men with CP/CPPS have more tenderness compared to men without CPPS. Tenderness in men with CPPS is distributed throughout the pelvis and not specific to the prostate. PMID- 17908329 TI - Morphine reduces local cytokine expression and neutrophil infiltration after incision. AB - BACKGROUND: Inflammation and nociceptive sensitization are hallmarks of tissue surrounding surgical incisions. Recent studies demonstrate that several cytokines may participate in the enhancement of nociception near these wounds. Since opioids like morphine interact with neutrophils and other immunocytes, it is possible that morphine exerts some of its antinociceptive action after surgical incision by altering the vigor of the inflammatory response. On the other hand, keratinocytes also express opioid receptors and have the capacity to produce cytokines after injury. Our studies were directed towards determining if opioids alter cytokine production near incisions and to identify cell populations responsible for producing these cytokines. RESULTS: A murine incisional model was used to measure the effects of acute morphine administration (0.1-10 mg/kg) on nociceptive thresholds, neutrophil infiltration and cytokine production in hind paw skin 30 minutes and 2 hours after incision. Incised hind paws displayed profound allodynia which was reduced by morphine (0.1-10 mg/kg) in the 2 hours following incision. Skin samples harvested from these mice showed enhanced levels of 5 cytokines: IL-1 beta, IL-6, tumor necrosis factor alpha (TNFalpha), granulocyte colony stimulating factor (G-CSF) and keratinocyte-derived cytokine (KC). Morphine reduced these incision-stimulated levels. Separate analyses measuring myeloperoxidase (MPO) and using immunohistochemistry demonstrated that morphine dose-dependently reduced the infiltration of neutrophils into the peri incisional tissue. The dose of morphine required for reduction of cytokine accumulation, however, was below that required for inhibition of peri-incisional neutrophil infiltration. Additional immunohistochemical studies revealed wound edge keratinocytes as being an important source of cytokines in the acute phase after incision. CONCLUSION: Acute morphine administration of doses as low as 0.1 mg/kg reduces peri-incisional cytokine expression. A reduction in neutrophil infiltration does not provide a complete explanation for this effect, and keratinocytes may be responsible for some incision area cytokine production. These studies suggest that morphine may alter the inflammatory milieu of incisional wounds, but these alterations do not likely contribute significantly to analgesia in the acute setting. PMID- 17908333 TI - Bioactive peptide design using the Resonant Recognition Model. AB - With a large number of DNA and protein sequences already known, the crucial question is to find out how the biological function of these macromolecules is "written" in the sequence of nucleotides or amino acids. Biological processes in any living organism are based on selective interactions between particular bio molecules, mostly proteins. The rules governing the coding of a protein's biological function, i.e. its ability to selectively interact with other molecules, are still not elucidated. In addition, with the rapid accumulation of databases of protein primary structures, there is an urgent need for theoretical approaches that are capable of analysing protein structure-function relationships. The Resonant Recognition Model (RRM) 12 is one attempt to identify the selectivity of protein interactions within the amino acid sequence. The RRM 12 is a physico-mathematical approach that interprets protein sequence linear information using digital signal processing methods. In the RRM the protein primary structure is represented as a numerical series by assigning to each amino acid in the sequence a physical parameter value relevant to the protein's biological activity. The RRM concept is based on the finding that there is a significant correlation between spectra of the numerical presentation of amino acids and their biological activity. Once the characteristic frequency for a particular protein function/interaction is identified, it is possible then to utilize the RRM approach to predict the amino acids in the protein sequence, which predominantly contribute to this frequency and thus, to the observed function, as well as to design de novo peptides having the desired periodicities. As was shown in our previous studies of fibroblast growth factor (FGF) peptidic antagonists 23 and human immunodeficiency virus (HIV) envelope agonists 24, such de novo designed peptides express desired biological function. This study utilises the RRM computational approach to the analysis of oncogene and proto oncogene proteins. The results obtained have shown that the RRM is capable of identifying the differences between the oncogenic and proto-oncogenic proteins with the possibility of identifying the "cancer-causing" features within their protein primary structure. In addition, the rational design of bioactive peptide analogues displaying oncogenic or proto-oncogenic-like activity is presented here. PMID- 17908334 TI - Why nonlinear biomedical physics? AB - The two goals of Nonlinear Biomedical Physics are: firstly to show how nonlinear methods can shed new light on biological phenomena and medical applications and secondly to bridge the technical, mathematical, and cultural divides between the physical disciplines where these methods are being developed and the audience for their use in the biological and medical sciences. PMID- 17908332 TI - Lipoprotein(a): an independent risk factor for ischemic heart disease that is dependent on triglycerides in subjects with type 2 diabetes mellitus. AB - Lipoprotein(a) is an independent risk factor for Ischaemic Heart Disease (IHD) in the general population. There are conflicting reports in the extent of its association with IHD among subjects with Type 2 diabetes mellitus (T2DM). The aim was to determine the concentration of Lp(a) and its relationship with other lipids parameters among Omani T2DM subjects with and without IHD. An over-night fasting blood sample from 221 T2DM subjects (86 females and 135 males) and 156 non-diabetics (69 females and 87 males) aged 30-70 years (as control) was taken for lipid profile studies. RESULTS: Lp(a) was significantly lower (p = 0.012) among T2DM subjects 0.123(1.12) g/L compared to non-diabetics 0.246 (1.18)g/L, irrespective of gender.A significant correlation (Spearman correlation, P = 0.047) was revealed between Lp(a) and IHD among Omani T2DM subjects. The proportions of T2DM subjects with IHD and an Lp(a) >0.3 g/L was higher compared to T2DM without IHD irrespective of gender, for women 42% vs. 27% and for men 17.5 vs. 8%, respectively.A significant negative correlation existed between Lp(a) and triglycerides (r = 0.41, P = 0.002) among T2DM subjects. In contrast, a significant positive correlation existed between Lp(a) and LDL-chol among the non diabetic subjects. Women had significantly higher Lp(a) concentration compared to men ( 0.30 Vs. 0.16 g/L, P < 0.0001) irrespective of the diabetic status. CONCLUSION: Lp(a) is an independent risk factor for IHD among Omani T2DM subjects. Lp(a) concentration was significantly lower and negatively correlated with triglycerides among Omani diabetic compared to non-diabetic subjects. PMID- 17908335 TI - Synchronized dynamics of cortical neurons with time-delay feedback. AB - The dynamics of three mutually coupled cortical neurons with time delays in the coupling are explored numerically and analytically. The neurons are coupled in a line, with the middle neuron sending a somewhat stronger projection to the outer neurons than the feedback it receives, to model for instance the relay of a signal from primary to higher cortical areas. For a given coupling architecture, the delays introduce correlations in the time series at the time-scale of the delay. It was found that the middle neuron leads the outer ones by the delay time, while the outer neurons are synchronized with zero lag times. Synchronization is found to be highly dependent on the synaptic time constant, with faster synapses increasing both the degree of synchronization and the firing rate. Analysis shows that pre-synaptic input during the inter-spike interval stabilizes the synchronous state, even for arbitrarily weak coupling, and independent of the initial phase. The finding may be of significance to synchronization of large groups of cells in the cortex that are spatially distanced from each other. PMID- 17908337 TI - Journal of the International Society of Sports Nutrition: a new era begins. PMID- 17908336 TI - Graph theoretical analysis of complex networks in the brain. AB - Since the discovery of small-world and scale-free networks the study of complex systems from a network perspective has taken an enormous flight. In recent years many important properties of complex networks have been delineated. In particular, significant progress has been made in understanding the relationship between the structural properties of networks and the nature of dynamics taking place on these networks. For instance, the 'synchronizability' of complex networks of coupled oscillators can be determined by graph spectral analysis. These developments in the theory of complex networks have inspired new applications in the field of neuroscience. Graph analysis has been used in the study of models of neural networks, anatomical connectivity, and functional connectivity based upon fMRI, EEG and MEG. These studies suggest that the human brain can be modelled as a complex network, and may have a small-world structure both at the level of anatomical as well as functional connectivity. This small world structure is hypothesized to reflect an optimal situation associated with rapid synchronization and information transfer, minimal wiring costs, as well as a balance between local processing and global integration. The topological structure of functional networks is probably restrained by genetic and anatomical factors, but can be modified during tasks. There is also increasing evidence that various types of brain disease such as Alzheimer's disease, schizophrenia, brain tumours and epilepsy may be associated with deviations of the functional network topology from the optimal small-world pattern. PMID- 17908339 TI - Virtual respiratory system in investigation of CPAP influence on optimal breathing frequency in obstructive lungs disease. AB - BACKGROUND: Continuous Positive Airway Pressure (CPAP) is a commonly accepted method of spontaneous breathing support in obstructive lung disease. Previous work suggested that the cause of the CPAP efficacy in the obstructive lung disease localized in bronchi of middle order (OLDMO) is not as obvious as, for example, in the obstructive sleep apnea. Since CPAP reduces obstruction and the optimal breathing frequency (BF) depends on the obstruction level, it seems to be important to analyze the dependence of the optimal BF on CPAP. AIM: To analyze the support efficacy cause in OLDMO, esp. the relationship between the CPAP value and optimal BF. METHOD: Investigations utilized previously built virtual respiratory system. Its most important factors: nonlinear lungs compliance and changeability of nonlinear airway resistance (Raw). Influence of BF and the CPAP value on the tidal volume and minute ventilation was analyzed for four exemplary virtual patients: healthy ("standard") and suffering from moderate, severe, and the very severe OLDMO (the other parameters, esp. respiratory muscles effort, were unchanged). Minute inspiratory work as a criterion of the BF optimization. RESULTS: CPAP decreased Raw making breathing easier, however, it shifted the working point of the respiratory system towards the smaller lungs compliance making breathing harder. The final result depended on the Raw value: CPAP improved breathing of patients with the serious OLDMO while it worsened healthy person breathing. The optimal CPAP value depended on the Raw value. If a virtual patient suffering from the serious OLDMO was not supported with CPAP, he had to breathe with low frequency because minute ventilation did not rise with BF increase. The optimal BF depended on the CPAP value (the greater the value, the greater the frequency). CONCLUSION: The CPAP efficacy depends on the level of OLDMO. CPAP is efficient in the severe OLDMO because it increases the optimal BF, which makes possible less energy-consuming breathing with frequency close to the normal one (greater BF means smaller tidal volume and thus smaller work against lungs compliance). PMID- 17908340 TI - Estimating the distribution of dynamic invariants: illustrated with an application to human photo-plethysmographic time series. AB - Dynamic invariants are often estimated from experimental time series with the aim of differentiating between different physical states in the underlying system. The most popular schemes for estimating dynamic invariants are capable of estimating confidence intervals, however, such confidence intervals do not reflect variability in the underlying dynamics. We propose a surrogate based method to estimate the expected distribution of values under the null hypothesis that the underlying deterministic dynamics are stationary. We demonstrate the application of this method by considering four recordings of human pulse waveforms in differing physiological states and show that correlation dimension and entropy are insufficient to differentiate between these states. In contrast, algorithmic complexity can clearly differentiate between all four rhythms. PMID- 17908338 TI - Effect of protein source and resistance training on body composition and sex hormones. AB - BACKGROUND: Evidence suggests an inverse relationship between soy protein intake and serum concentrations of male sex hormones. Anecdotal evidence indicates that these alterations in serum sex hormones may attenuate changes in lean body mass following resistance training. However, little empirical data exists regarding the effects of soy and milk-based proteins on circulating androgens and exercise induced body composition changes. METHODS: For 12 weeks 20 subjects were supplemented with 50 g per day of one of four different protein sources (Soy concentrate; Soy isolate; Soy isolate and whey blend, and Whey blend only) in combination with a resistance-training program. Body composition, testosterone, estradiol and sex hormone binding globulin (SHBG) were measured at baseline and week 12. RESULTS: Protein supplementation resulted in a significant increase in lean body mass independent of protein source (0.5 +/- 1.1 and 0.9 +/- 1.4 kg, p = 0.006, p = 0.007). No significant differences were observed between groups for total and free testosterone, SHBG, percentage body fat, BMI or body weight. The Testosterone/Estradiol ratio increased across all groups (+13.4, p = 0.005) and estradiol decreased (p = 0.002). Within group analysis showed significant increases in the Testosterone/Estradiol ratio in soy isolate + whey blend group (+16.3, p = 0.030). Estradiol was significantly lower in the whey blend group ( 9.1 +/- 8.7 pg/ml, p = 0.033). CONCLUSION: This investigation shows that 12 week supplementation with soy protein does not decrease serum testosterone or inhibit lean body mass changes in subjects engaged in a resistance exercise program. PMID- 17908341 TI - Stochastic nonlinear dynamics pattern formation and growth models. AB - Stochastic evolutionary growth and pattern formation models are treated in a unified way in terms of algorithmic models of nonlinear dynamic systems with feedback built of a standard set of signal processing units. A number of concrete models is described and illustrated by numerous examples of artificially generated patterns that closely imitate wide variety of patterns found in the nature. PMID- 17908342 TI - Phospholipids and sports performance. AB - Phospholipids are essential components of all biological membranes. Phosphatidylcholine (PC) and Phosphatidylserine (PS) are Phosphatidyl phospholipids that are required for normal cellular structure and function. The participation in physical activity often challenges a variety of physiological systems; consequently, the ability to maintain normal cellular function during activity can determine sporting performance. The participation in prolonged intense exercise has been shown to reduce circulatory choline concentrations in some individuals. As choline is a pre-cursor to the neurotransmitter Acetylcholine, this finding has encouraged researchers to investigate the hypothesis that supplementation with PC (or choline salts) could enhance sporting performance. Although the available data that evaluates the effects of PC supplementation on performance are equivocal, acute oral supplementation with PC (~0.2 g PC per kg body mass) has been demonstrated to improve performance in a variety of sporting activities where exercise has depleted circulatory choline concentrations. Short term oral supplementation with soy-derived PS (S-PS) has been reported to attenuate circulating cortisol concentrations, improve perceived well-being, and reduce perceived muscle soreness after exercise. More recently, short term oral supplementation (750 mg per day of S-PS for 10 days) has been demonstrated to improve exercise capacity during high intensity cycling and tended to increase performance during intermittent running. Although more research is warranted to determine minimum dietary Phospholipid requirements for optimal sporting performance, these findings suggest that some participants might benefit from dietary interventions that increase the intakes of PC and PS. PMID- 17908344 TI - From conformons to human brains: an informal overview of nonlinear dynamics and its applications in biomedicine. AB - Methods of contemporary physics are increasingly important for biomedical research but, for a multitude of diverse reasons, most practitioners of biomedicine lack access to a comprehensive knowledge of these modern methodologies. This paper is an attempt to describe nonlinear dynamics and its methods in a way that could be read and understood by biomedical professionals who usually are not trained in advanced mathematics. After an overview of basic concepts and vocabulary of nonlinear dynamics, deterministic chaos, and fractals, application of nonlinear methods of biosignal analysis is discussed. In particular, five case studies are presented: 1. Monitoring the depth of anaesthesia and of sedation; 2. Bright Light Therapy and Seasonal Affective Disorder; 3. Analysis of posturographic signals; 4. Evoked EEG and photo stimulation; 5. Influence of electromagnetic fields generated by cellular phones. PMID- 17908343 TI - A novel aromatic oil compound inhibits microbial overgrowth on feet: a case study. AB - BACKGROUND: Athlete's Foot (Tinea pedis) is a form of ringworm associated with highly contagious yeast-fungi colonies, although they look like bacteria. Foot bacteria overgrowth produces a harmless pungent odor, however, uncontrolled proliferation of yeast-fungi produces small vesicles, fissures, scaling, and maceration with eroded areas between the toes and the plantar surface of the foot, resulting in intense itching, blisters, and cracking. Painful microbial foot infection may prevent athletic participation. Keeping the feet clean and dry with the toenails trimmed reduces the incidence of skin disease of the feet. Wearing sandals in locker and shower rooms prevents intimate contact with the infecting organisms and alleviates most foot-sensitive infections. Enclosing feet in socks and shoes generates a moisture-rich environment that stimulates overgrowth of pungent both aerobic bacteria and infectious yeast-fungi. Suppression of microbial growth may be accomplished by exposing the feet to air to enhance evaporation to reduce moistures' growth-stimulating effect and is often neglected. There is an association between yeast-fungi overgrowths and disabling foot infections. Potent agents virtually exterminate some microbial growth, but the inevitable presence of infection under the nails predicts future infection. Topical antibiotics present a potent approach with the ideal agent being one that removes moisture producing antibacterial-antifungal activity. Severe infection may require costly prescription drugs, salves, and repeated treatment. METHODS: A 63-y female volunteered to enclose feet in shoes and socks for 48 hours. Aerobic bacteria and yeast-fungi counts were determined by swab sample incubation technique (1) after 48-hours feet enclosure, (2) after washing feet, and (3) after 8-hours socks-shoes exposure to a aromatic oil powder compound consisting of arrowroot, baking soda, basil oil, tea tree oil, sage oil, and clove oil. CONCLUSION: Application of this novel compound to the external surfaces of feet completely inhibited both aerobic bacteria and yeast-fungi-mold proliferation for 8-hours in spite of being in an enclosed environment compatible to microbial proliferation. Whether topical application of this compound prevents microbial infections in larger populations is not known. This calls for more research collected from subjects exposed to elements that may increase the risk of microbial-induced foot diseases. PMID- 17908345 TI - Is there a difference in quality of life, comparing summer and winter, in patients who have undergone total laryngectomy? AB - INTRODUCTION: Several studies have implied that patients' quality of life stabilises six months after undergoing total laryngectomy. However, these studies may well have overlooked persistent short term variations in patients' quality of life. The aim of this study was to assess the impact of seasonal change (i.e. summer vs winter) on the quality of life of patients following total laryngectomy. METHODS: A prospective, cross-sectional study of recurrence-free laryngectomy patients was performed, using the European Organization for Research and Treatment of Cancer (version three) QLQ-C30 questionnaires (EORTC QLQ-C30), during January 2004 (i.e. winter), August 2004 (summer) and January 2005 (winter). RESULTS: Thirty-six patients were entered into the study. The response rate was 70.3 per cent. Patient's time elapsed since surgery varied from six months to 12 years. In all questionnaire domains, responses seemed remarkably consistent over time. CONCLUSION: No statistically significant differences were found between summer and winter scores in all the domains analysed by the EORTC QLQ-C30. PMID- 17908346 TI - Improving correspondence to general practitioners regarding patients attending the ENT emergency clinic: a regional general practitioner survey and audit. AB - INTRODUCTION: The ENT emergency clinic provides an important out-patient service; however, often, no correspondence is sent to the general practitioner. AIM: To conduct a general practitioner questionnaire audit in order to assess whether a standardised, computerised clinic letter template could improve communication between the ENT emergency clinic and patients' general practitioners. STANDARD: All ENT emergency clinic patients should have a summary letter sent to their general practitioner. METHODS: One hundred general practitioner questionnaires were enclosed with the first 100 ENT emergency clinic template letters sent to patients' general practitioners. RESULTS: Seventy-two general practitioners responded (72 per cent). Of these respondents, only 7 per cent had previously received regular correspondence from the ENT emergency clinic before the introduction of the computerised letter template. Following its implementation, such a letter was sent to 100 per cent of the clinic patients' general practitioners. Ninety-seven per cent of the general practitioners valued the template letter, with a mean satisfaction score of 8.4 on a 10-point scale. Eighty-six per cent of the general practitioners stated that they would not prefer a dictated letter. CONCLUSION: The introduction of a simple, computerised clinic letter template improves communication with ENT emergency clinic patients' general practitioners. PMID- 17908347 TI - Health services use for mental health problems by community-living seniors with depression. AB - BACKGROUND: The literature on health services (HS) use for mental health problems by community-living seniors with depression is sparse. This study aimed to characterize patterns of HS use for mental health problems by seniors with depressive disorders and symptoms during the previous 12 months. METHOD: The study used data from the Canadian Community Health Survey Cycle 1.2: Mental Health and Well-Being (N = 7736). Information was collected on demographic, social, mental, physical and functional variables and HS use for mental health problems. We obtained simple univariate and bivariate descriptions of the data and developed multivariate predictive models for each of the HS utilization variables. RESULTS: Rates of any HS use for mental health problems ranged from 1.8% for those with no depressive symptoms to 31.1% for those with major depression. Variables predicting increased HS use were: depressive disorder or symptoms, clinically significant distress or impairment, age 65-84, single, post secondary education, religiousness, disability, co-morbid mental disorder and fewer friends and positive social interactions. Variables predicting HS use among depressed seniors were physical health, psychiatric co-morbidity and activity limitation. CONCLUSION: Community-living seniors with major depression, co-morbid major depression and depressive symptoms were more likely to use psychiatrist, family physician, other professional and self-help services for mental health problems but less than a third actually did so. PMID- 17908348 TI - Time that tells: critical clock-drawing errors for dementia screening. AB - BACKGROUND: Clock-drawing tests are popular components of dementia screens but no single scoring system has been universally accepted. We sought to identify an optimal subset of clock errors for dementia screening and compare them with three other systems representative of the existing wide variations in approach (Shulman, Mendez, Wolf-Klein), as well as with the CDT system used in the Mini Cog, which combines clock drawing with delayed recall. METHODS: The clock drawings of an ethnolinguistically and educationally diverse sample (N = 536) were analyzed for the association of 24 different errors with the presence and severity of dementia defined by independent research criteria. The final sample included 364 subjects with > or = 5 years of education, as preliminary examination suggested different error patterns in subjects with 0-4 years of education and inadequate numbers of normal controls for reliable analysis. RESULTS: Eleven of 24 errors were significantly associated with dementia in subjects with > or = 5 years of education, and six were combined to identify dementia with 88% specificity and 71% sensitivity: inaccurate time setting, no hands, missing numbers, number substitutions or repetitions, or refusal to attempt clock drawing. Time setting was the most prevalent error at all dementia stages, refusal occurred only in moderate and severe dementia; and ethnicity and language of administration had no effect. All critical errors increased in frequency with dementia stage. This simplified scoring system had much better specificity than two other systems (88% vs 39% for Mendez's system - 63% for Shulman's) and much better sensitivity than Wolf-Klein's (71% vs 51%). Stepwise logistic regression found the simplified system to be more strongly predictive of dementia than the three other CDT systems of dementia. Substituting the new CDT algorithm for that used in the original CDT Mini-Cog improved the Mini-Cog's specificity from 89 to 93% with minimal change in sensitivity. CONCLUSIONS: Only six errors need be assessed to capture most of the power of clock drawing to discriminate between people with dementia and normal subjects, and improves specificity over older systems in subjects with > or = 5 years of education. These errors require minimal conceptual classification and are easily detected and scored by non-specialists. PMID- 17908350 TI - Vestibular signs and symptoms of volumetric abnormalities of the vestibular aqueduct. AB - OBJECTIVE: The objective of this study was to identify a pattern of signs, symptoms and neuroradiological findings which would assist investigation of vestibular function (especially otolith function) in a group of adult patients with anatomical alterations of the endolymphatic aqueduct and sac. METHODS: Fifteen subjects affected by volumetric abnormalities of the vestibular aqueduct were selected from a cohort of patients referred to a tertiary referral neurotological centre between 1 January 2004 and 30 June 2006. All patients underwent accurate clinical history-taking and were evaluated using a standardised set of bedside and instrumental neurotological tests (i.e. audiometry, auditory brainstem response and vestibular evoked myogenic potentials). After these tests, each patient underwent computed tomography and magnetic resonance imaging in order to accurately evaluate the middle ear, labyrinthine capsule and internal auditory canals. These evaluations confirmed clinical suspicion of volumetric abnormalities of the vestibular aqueduct and endolymphatic sac. RESULTS: All the patients with a defined volumetric alteration in the region of the vestibular aqueduct and endolympatic sac reported a typical pattern of symptoms and signs. The most obvious and frequent symptoms in these patients were migraine-related vertigo (using the Neuhauser criteria, 10 of 15, 66.6 per cent), 'motion sickness' (12 of 15, 80 per cent), oscillopsia (nine of 15, 60 per cent) and dizziness (14 of 15, 93.3 per cent). Clinical examination results for the selected patients allowed some useful speculative conclusions. During neurotological evaluation, two instrumental methodologies were especially useful diagnostically: vestibular evoked myogenic potentials of the neck, and the mastoid vibration test at 100 Hz. CONCLUSIONS: Dysfunction of the vestibular aqueduct is suggested by symptomatology characterised by: migraine-related vertigo, unstable or recurring oscillopsia, lowering of the vestibular evoked myogenic potential threshold, hypoacusis, anamnestic report of motion sickness, and nystagmus induced by mastoid vibration and head-shaking. Computed tomography and magnetic resonance imaging are needed in order to confirm clinical suspicions. PMID- 17908349 TI - Mechanism underlying rebound excitation in retinal ganglion cells. AB - Retinal ganglion cells (RGCs) display the phenomenon of rebound excitation, which is observed as rebound sodium action potential firing initiated at the termination of a sustained hyperpolarization below the resting membrane potential (RMP). Rebound impulse firing, in contrast to corresponding firing elicited from rest, displayed a lower net voltage threshold, shorter latency and was invariably observed as a phasic burst-like doublet of spikes. The preceding hyperpolarization leads to the recruitment of a Tetrodotoxin-insensitive depolarizing voltage overshoot, termed as the net depolarizing overshoot (NDO). Based on pharmacological sensitivities, we provide evidence that the NDO is composed of two independent but interacting components, including (1) a regenerative low threshold calcium spike (LTCS) and (2) a non-regenerative overshoot (NRO). Using voltage and current clamp recordings, we demonstrate that amphibian RGCs possess the hyperpolarization activated mixed cation channels/current, Ih, and low voltage activated (LVA) calcium channels, which underlie the generation of the NRO and LTCS respectively. At the RMP, the Ih channels are closed and the LVA calcium channels are inactivated. A hyperpolarization of sufficient magnitude and duration activates Ih and removes the inactivation of the LVA calcium channels. On termination of the hyperpolarizing influence, Ih adds an immediate depolarizing influence that boosts the generation of the LTCS. The concerted action of both conductances results in a larger amplitude and shorter latency NDO than either mechanism could achieve on its own. The NDO boosts the generation of conventional sodium spikes which are triggered on its upstroke and crest, thus eliciting rebound excitation. PMID- 17908352 TI - Primary B-cell lymphoma presenting as bilateral ear lobule swelling. AB - We report a rare case of primary B-cell lymphoma presenting as bilateral ear lobule swelling. A 56-year-old white man presented with a one-year history of painless swelling of both ear lobules. An excision biopsy confirmed B-cell lymphoma. Detailed systemic investigation confirmed the primary nature of the tumour. This tumour is rare in the ear lobule. A review of the English literature revealed no previously reported case of bilateral primary ear lobule involvement. Clinicians should be aware that this tumour can present as a primary in the ear lobules. PMID- 17908351 TI - Time trend analysis of otological procedures performed in England, 1989 to 2005. AB - OBJECTIVE: To observe trends in the number of major otological procedures performed in England, in the context of advances in the understanding of disease. METHODS: The data used were obtained from the Hospital Episode Statistics statistical database, published by the UK Department of Health, for England, 1989 to 2005. Specific otological procedures were identified using the Classification of Surgical Operations and Procedures system (fourth revision) of the Office of Population, Censuses and Surveys. Trend analysis of different procedures was performed using exponential smoothing (using the Statistical Package for the Social Sciences version 13 software). RESULTS: Our study did not confirm any reduction in the number of surgical procedures performed for cholesteatoma or otosclerosis. We noted a sharp decline in the number of endolymphatic sac surgical procedures performed, probably attributable to the increased use of intratympanic therapy. CONCLUSION: The number of major otological procedures (other than endolymphatic sac surgery) was consistent over the period examined. The generally perceived reduction in the number of procedures performed by individual surgeons may be due to a dilutional effect. This can only support the need for subspecialisation, particularly regarding the training of junior surgeons. PMID- 17908354 TI - Botulinum toxin as adjunctive therapy in refractory laryngeal granuloma. AB - OBJECTIVE: This study evaluated the role of botulinum toxin type A in the treatment of refractory laryngeal granulomas. STUDY DESIGN AND SETTING: Retrospective clinical review at a tertiary care hospital. Seven patients with vocal process granulomas underwent percutaneous injection of botulinum toxin into both vocal folds, performed in an office setting. Total doses ranged from 10 to 25 U, divided between both vocal folds. RESULTS: All patients experienced resolution of their granulomas over two to seven weeks. No patient developed aspiration pneumonia. All patients experienced hoarseness secondary to the injections, but voice quality returned to baseline in all patients as the toxin was degraded. CONCLUSIONS: Botulinum toxin is safe and effective therapy in resolving vocal process granulomas in patients refractory to traditional therapy. The optimal treatment dose remains to be determined. SIGNIFICANCE: Percutaneous botulinum toxin injection is helpful in resolving laryngeal granulomas. PMID- 17908353 TI - Dual silencing of type 1 insulin-like growth factor and epidermal growth factor receptors to induce apoptosis of nasopharyngeal cancer cells. AB - OBJECTIVE: The epidermal growth factor receptor (EGFR) and type 1 insulin-like growth factor receptor (type 1 IGF receptor or IGF1R) have played an important role in the growth and apoptosis of cancer. The RNA interference (RNAi) technique can suppress gene expression, but the effects of dual silencing of EGFR and type 1 IGF receptor have not been well understood. METHODS: pU6-EGFR-shRNA-1, pU6-EGFR shRNA-2, pU6-IGF1R-shRNA-1 and pU6-IGF-1R-shRNA-2 plasimd vectors were transfected to the nasopharyngeal cancer cells. Seven groups were selected for the study. The protein and downstream protein expression were assessed by Western blot. Apoptosis was determined via flow cytometry. Meanwhile, chemosensitivity of nasopharyngeal cancer cell lines transfeced to chemotherapeutic drugs were carried out by MTT. RESULTS: In dual silencing of EGFR and IGF-1R, the protein expression much more was decreased than single silencing of EGFR or IGF-1R, but the cell apoptosis much more is increased than single silencing EGFR or IGF-1R. Dual silencing of EGFR and IGF-1R enhanced chemosensitivity to anticancer drugs, compared with single silencing of EGFR or IGF-1R. CONCLUSION: Dual silencing of EGFR and IGF-1R are capable of suppressing EGFR and IGF-1R expression of the nasopharyngeal cancer cell and can promote apoptosis and increase the cell sensitivity of anticancer drug. The dual silencing of genes RNAi technique is significantly better than a single gene. PMID- 17908355 TI - Gorlin's syndrome presenting with myolipoma of tongue base. AB - OBJECTIVE: We report the first case of tongue base myolipoma associated with Gorlin's syndrome. METHOD: Case report and review of world literature. RESULTS: A 39-year-old man with known Gorlin's syndrome presented with progressive dysphagia. Subsequent magnetic resonance imaging scan and biopsy confirmed the rare diagnosis of myolipoma arising from the tongue base. In view of the benign nature of this mass, it was debulked rather than completely excised in order to preserve swallowing function. CONCLUSION: To our knowledge, this is the first report of the coexistence of two rare conditions in an atypical fashion. This case appears to represent a new variant in the broad spectrum of features of Gorlin's syndrome. PMID- 17908356 TI - No evidence of linkage between 7q33-36 locus (OTSC2) and otosclerosis in seven British Caucasian pedigrees. AB - BACKGROUND: The aetiology of otosclerosis is complex, and probably involves an interaction between genes and environmental factors. Previous studies have revealed genetic linkage with a number of chromosome regions, including position 7q33-36. AIM: To confirm whether linkage exists between otosclerosis and chromosome region 7q33-36. MATERIALS AND METHODS: Seven multiply affected families were ascertained. Deoxyribonucleic acid from members of these families was extracted, and six markers were genotyped to cover a 16 cM region at 7q33-36. Both parametric and non-parametric multipoint linkage analyses were performed. RESULTS: Parametric multipoint linkage analysis excluded any linkage at 7q33-36 (logarithm of odds score <-4.0). Non-parametric linkage analysis also failed to confirm any linkage (non-parametric linkage < 1.66). When tested individually, pedigree four was the only one to show a significant non-parametric linkage score between D7s684 and D7s2513 (non-parametric linkage = 1.96). CONCLUSION: No linkage was detected between otosclerosis and the 7q33-36 region. This could be explained by the study's lack of power, due to the limited number of families available. PMID- 17908357 TI - Diagnostic tests for immunomediated hearing loss: a systematic review. AB - OBJECTIVE: To quantitatively evaluate the diagnostic accuracy of diagnostic tests for immunomediated hearing loss. DATA SOURCES: We searched Medline and the Cochrane Database of Systematic Reviews for potentially relevant studies. STUDY SELECTION: Twenty-five studies met the inclusion criteria of this systematic review. The diagnosis of immunomediated hearing loss was based on the clinical presentation and the response to corticosteroid administration. DATA EXTRACTION: The following data were extracted from the selected studies and entered into a standardised database: population demographics; exclusion and inclusion criteria; diagnostic tests; sensitivity; specificity; the number of true positive, true negative, false positive and false negative values; therapy used, including dose and duration; and delay between symptom onset and therapy commencement. DATA SYNTHESIS: This systematic review combined data from 679 patients with immunomediated hearing loss, reported by 22 research teams. Substantial heterogeneity was found among the included studies; for this reason, summary sensitivity and specificity values were not computed. CONCLUSIONS: The results of diagnostic tests for immunomediated hearing loss depend on many factors, and there is a risk of potential bias. This is the first time that such a systematic review has been presented; such a review is a more rigorous method of demonstrating the utility of the available diagnostic tests. PMID- 17908358 TI - Chondrosarcoma of larynx: review of literature and clinical experience. AB - Laryngeal chondrosarcomas are rare, cartilaginous tumours, and around 300 cases have been reported in the literature. They are slow-growing tumours which present difficulties both in diagnosis and treatment. Most patients eventually require radical surgery. However, over the last 10 years, we have diagnosed two cases of laryngeal chondrosarcoma and managed them conservatively by endoscopic debulking and regular follow up. The objective of this article is to highlight the conservative approach in managing these patients, particularly in the presence of co-morbid conditions. PMID- 17908359 TI - Heterozygosity and parasite intensity: lung parasites in the water frog hybridization complex. AB - In hybridogenetic systems, hybrid individuals are fully heterozygous because one of the parental genomes is discarded from the germinal line before meiosis. Such systems offer the opportunity to investigate the influence of heterozygosity on susceptibility to parasites. We studied the intensity of lung parasites (the roundworm Rhabdias bufomis and the fluke Haplometra cylindracea) in 3 populations of water frogs of the Rana lessonae-esculenta complex in eastern France. In these mixed populations, hybrid frogs (R. esculenta) outnumbered parental ones (R. lessonae). Despite variation in parasite intensity and demographic variability among populations, the relationship between host age and intensity of parasitism suggests a higher susceptibility in parentals than in hybrids. Mortality is probably enhanced by lung parasites in parental frogs. On the other hand, while parental frogs harboured higher numbers of H. cylindracea than hybrid frogs, the latter had higher numbers of R. bufonis. Despite such discrepancies, these results support the hybrid resistance hypothesis, although other factors, such as differences in body size, age-related immunity, differential exposure risks and hemiclonal selection, could also contribute to the observed patterns of infection. PMID- 17908360 TI - Mechanisms underlying reduced expulsion of a murine nematode infection during protein deficiency. AB - Balb/c mice infected with the gastrointestinal nematode Heligmosomoides bakeri were fed protein sufficient (PS, 24%) or deficient (PD, 3%) diets to investigate whether diet, infection or dose of larval challenge (0, 100 or 200 larvae) influenced gut pathophysiology and inflammation. Among the PS mice, worms were more posteriorad in the intestine of mice infected with 200 compared with 100 larvae, suggesting active expulsion in the more heavily infected mice. This was consistent with the positive correlation between worm numbers and fluid leakage in PS mice; similar patterns were not detected in the PD mice. Infection also induced villus atrophy, which was more pronounced in PS than in PD mice. Our cytokine screening array indicated that infection in PD mice elevated a wide range of pro-inflammatory cytokines and chemokines. Whereas serum leptin concentrations were higher in PD mice, monocyte chemotactic protein-5 (MCP-5) in serum increased with increasing larval dose and concentrations were lower in PD than PS mice. We suggest that elevated MCP-5 together with villus atrophy may contribute to the apparent dose-dependent expulsion of H. bakeri from PS mice but that delayed expulsion in PD mice appeared related to a predominant Th1 cytokine profile that may be driven by leptin. PMID- 17908361 TI - Vesicle trafficking during sporozoite development in Plasmodium berghei: ultrastructural evidence for a novel trafficking mechanism. AB - Oocysts from Anopheles stephensi mosquitoes fed on murine blood infected with Plasmodium berghei berghei, were fixed for electron microscopy 6-12 days post feeding. Ultrastructural analysis focused on Golgi-related trafficking pathways for rhoptry and microneme formation during sporogony. A small Golgi complex of 1 3 cisternae is formed close to the spindle pole body from coated vesicles budded from the nuclear envelope which is confluent with the endoplasmic reticulum. Rhoptries begin as small spheroidal bodies apparently formed by fusion of Golgi derived vesicles, lengthening to 3-4 microm, and increasing in number to 4 per sporozoite. Ultrastructural data indicate the presence of a novel mechanism for vesicle transport between the Golgi complex and rhoptries along a longitudinal 30 nm - thick fibre (rootlet fibre or tigelle). Filamentous links between vesicles and rootlet indicate that this is a previously undescribed vesicle transport organelle. Genesis of micronemes occurs late in bud maturation and starts as spheroidal dense-cored vesicles (pro-micronemes), transforming to their mature bottle-like shape as they move apically. Filamentous links also occur between micronemes and subpellicular microtubules, indicating that as in merozoites, micronemes are trafficked actively along these structures. PMID- 17908362 TI - Searching for general patterns in parasite ecology: host identity versus environmental influence on gamasid mite assemblages in small mammals. AB - The abundance and diversity of parasites vary among different populations of host species. In some host-parasite associations, much of the variation seems to depend on the identity of the host species, whereas in other cases it is better explained by local environmental conditions. The few parasite taxa investigated to date make it difficult to discern any general pattern governing large-scale variation in abundance or diversity. Here, we test whether the abundance and diversity of gamasid mites parasitic on small mammals across different regions of the Palaearctic are determined mainly by host identity or by parameters of the abiotic environment. Using data from 42 host species from 26 distinct regions, we found that mite abundances on different populations of the same host species were more similar to each other than expected by chance, and varied significantly among host species, with half of the variance among samples explained by differences between host species. A similar but less pronounced pattern was observed for mite diversity, measured both as species richness and as the taxonomic distinctness of mite species within an assemblage. Strong environmental effects were also observed, with local temperature and precipitation correlating with mite abundance and species richness, respectively, across populations of the same host species, for many of the host species examined. These results are compared to those obtained for other groups of parasites, notably fleas, and discussed in light of attempts to find general rules governing the geographical variation in the abundance and diversity of parasite assemblages. PMID- 17908363 TI - Halfway up the trophic chain: development of parasite communities in the sparid fish Boops boops. AB - We examined the patterns of composition and structure of parasite communities in the Mediterranean sparid fish Boops boops along a gradient of fish sizes, using a large sample from a single population. We tested the hypothesis that species forming the core of the bogue parasite fauna (i.e. species which have a wide geographical range and are responsible for recognizable community structure) appear early in the fish ontogeny. The sequential community development observed supported the prediction that core species appear in the fish population earlier than rare and stochastic species. There was also a strong correlation between the order of 'arrival' of the species and their overall prevalence. Six key species were responsible for recognizable community structure across size/age cohorts; the addition to this baseline community of key parasite species resulted in a nested structure that is linked to differential species abundance rather than fish size. Information on the life-cycles, distribution and host range of the parasites is used to explain the observed patterns of parasite community structure. We conclude that the small mouth size of B. boops coupled with suction feeding may provide a setting for passive sampling as a mechanism leading to non random parasite community structure. PMID- 17908364 TI - Identification and characterization of a novel specific secreted protein family for selected members of the subfamily Ostertagiinae (Nematoda). AB - It has been shown that the bovine abomasal parasite, Ostertagia ostertagi, drastically modulates its microenvironment, causing epithelial cell damage, accumulation of inflammatory cells and pH changes in the stomach. The mechanisms used by the parasite to change the abomasal environment are largely unknown, but an important role has been attributed to excretory-secretory (ES) products from the parasite. In this study we have identified proteins representing a novel ES protein family, characterized by the SCP/Tpx-1/Ag5/PR-1/Sc7 protein motif. These proteins were named Oo-AL1 and Oo-AL2 (O. ostertagi ASP-like protein). Both proteins contain a signal peptide and 1 predicted N-glycosylation site. The transcript for Oo-AL1 was present from the L4 stage onwards in both male and female adult worms, whereas the Oo-AL2 transcript was hardly detectable. Western blots of somatic extracts and ES products from different developmental stages of O. ostertagi, probed with anti-Oo-AL1 antibodies, revealed Oo-AL proteins in the ES products of adult worms. An analysis of the nematode genome and EST databases indicated that these novel ES proteins are unique to O. ostertagi and its relative, Teladorsagia circumcincta, suggesting a key function in these abomasal parasites. PMID- 17908366 TI - Dietary zinc intake and brain cancer in adults: a case-control study. AB - Little is known about the aetiology of brain tumours. One putative factor suggested from animal models is a protective effect of dietary Zn. We tested the hypothesis that increased compared with low dietary Zn intake is protective against brain tumour development. We conducted a population-based case-control study in the UK, of adults aged 18-69 years, between 2001 and 2004 aiming to identify possible risk factors. Dietary information was collected from 637 cases diagnosed with a glioma or meningioma, and 876 controls. Data were obtained from a self-completed FFQ. Multivariate logistic regression analysis was conducted, adjusting for socio-demographic factors, season of questionnaire return, multivitamin supplementation and energy intake. Although a weak protective effect was observed for the third quartile of intake (normal compared with low intake) in the meningioma group, this was limited to the specific brain tumour subtype and quartile, and was not significant after also adjusting for intake of other elements. Overall there was no significant effect of Zn intake. No association or dose-response relationship was observed between increased compared with low Zn intake and risk of glioma or meningioma. PMID- 17908367 TI - Understanding the impact of prior depression on stress generation: examining the roles of current depressive symptoms and interpersonal behaviours. AB - Stress generation is a process in which individuals contribute to stressful life events. While research has supported an association between current depression and stress generation, it has been noted that individuals with prior depression tend to contribute to stressors even when they are no longer experiencing a depressive episode. The aim of the study is to elucidate the pathways through which prior major depression predicts interpersonal stress generation in women. Specifically, we examined current subsyndromal depressive symptoms and problematic interpersonal behaviours as potential mediators. Fifty-one college women were followed prospectively for 6 weeks. Participants were interviewed to assess current and past depression as well as stressful life events they experienced over the 6-week period. The findings suggest that prior major depression continues to have an impact even after the episode has ended, as the disorder continues to contribute to stress generation through residual depressive symptoms. PMID- 17908368 TI - Improved statistics for contrasting means of two samples under non-normality. AB - This paper presents the asymptotic expansions of the distributions of the two sample t-statistic and the Welch statistic, for testing the equality of the means of two independent populations under non-normality. Unlike other approaches, we obtain the null distributions in terms of the distribution and density functions of the standard normal variable up to n(-1), where n is the pooled sample size. Based on these expansions, monotone transformations are employed to remove the higher-order cumulant effect. We show that the new statistics can improve the precision of statistical inference to the level of o (n(-1)). Numerical studies are carried out to demonstrate the performance of the improved statistics. Some general rules for practitioners are also recommended. PMID- 17908369 TI - The psychometricians' fallacy: too clever by half? AB - The psychometricians' fallacy concludes that an attribute is quantitative from the premise that it is ordinal. This fallacy occupies a central place in the paradigm of psychometrics. Most of the founders of the discipline committed it and it makes sense of otherwise anomalous developments within the discipline, such as the permissible statistics controversy and the dominant form taken by item response theories. The fallacy is displayed by showing (1) that an attribute's quantitative structure reduces to a weak order upon differences between degrees that satisfies the double cancellation, solvability, and Archimedean conditions of conjoint measurement theory and (2) the fact that any order on the degrees themselves does not entail sufficient structure on this weak order to guarantee satisfaction of these conditions. Thus, it is possible that an ordered attribute is non-quantitative. Also, each pair of differences between degrees of an ordinal attribute falls into one of two disjoint classes: (1) those where the order relation between the pair follows from an order on the attribute and (2) those where it is independent of that order and possibly diagnostic of quantitative structure and this fact means that the distinction between order and quantity is an empirical one. PMID- 17908370 TI - Integrating an evidence-based assessment of benefit and risk in disease-modifying treatment of multiple sclerosis. AB - BACKGROUND: As results from an increasing number of clinical trials with disease modifying drugs (DMDs) in multiple sclerosis (MS) become available, the challenge for the treating neurologist is how to decide on the appropriate therapy for an individual patient. OBJECTIVE: An International Working Group for Treatment Optimization in MS met to consider how the principles of evidence-based medicine (EBM) should be used to assess the current best evidence regarding the treatment of MS. This report summarizes the outcome from the workshop at which this topic was addressed. RESULTS: Class I evidence from head-to-head studies provides the best tool for direct comparisons of DMDs. However, other EBM approaches to data analysis from placebo-controlled trials can be used to help determine the benefits and risks of a particular DMD relative to placebo by calculating the number needed to treat to achieve a positive outcome, such as avoiding a relapse, and the number needed to harm to produce an additional adverse event, such as having a therapy-related dropout. This provides a structured basis for comparisons between DMDs. CONCLUSION: While such comparisons have their limitations, particularly when drugs with substantially different side-effect profiles are to be compared, they can provide useful information to guide treatment decisions. PMID- 17908371 TI - An evaluation of the association between systemic inflammation--as measured by C reactive protein--and hospital resource use. AB - OBJECTIVE: To evaluate the association between inflammatory status, as measured by C-reactive protein (CRP), during inpatient admission and subsequent inpatient outcome and associated resource use. METHODS: Probabilistic record linkage was used to match hospital episode data, laboratory reports and mortality statistics in a large urban population of 424,000 people in South Wales, UK. Inpatient mortality, length of stay, emergency readmissions and subsequent 1-year hospital bed day occupancy were assessed as a function of CRP status. RESULTS: Between 2001 and 2005, in total there were 432,272 CRP observations from 98,505 people; 69,593 admissions had at least one CRP measurement, affecting 47,100 individual patients. Across all ICD-10 primary diagnoses, CRP was acutely high (> 10 mg/L) in three-quarters of admissions. Acutely high CRP was associated with an eight fold increase in risk of hospital mortality (p < 0.001) and a doubling of length of stay (p < 0.001) compared to normal CRP levels, after standardising for age and gender. Across the range of observed maximum CRP values measured during admissions (1 mg/L to > 400 mg/L) the likelihood of emergency readmission within 28 days of discharge increased by 50% (p < 0.001), and the predicted number of subsequent bed days occupied in the year following discharge increased by 30-58% across the range of CRP measurement (p = 0.004). CONCLUSIONS: CRP has been found to be clearly associated with hospital resource use. Furthermore, CRP also predicted in-hospital mortality. This may imply that better management of systemic inflammation would result in resource savings in inflammatory diseases such as rheumatoid arthritis. PMID- 17908372 TI - Relations among competence beliefs, utility value, achievement goals, and effort in mathematics. AB - BACKGROUND: Research has shown that motivation is a key factor in the learning process as well as in school achievement. In essence, a number of researchers have highlighted the close link between motivation and achievement-related behaviours such as effort. AIMS: The present study aims to acquire more specific information concerning the relations between competence beliefs, utility value and achievement goals in mathematics among secondary school students, to further document the influence of social agents, and to better understand the relationships between these variables, as well as to effort. SAMPLE: Participants were 759 Grade 7 to Grade 11 students (389 males, 370 females). METHOD: Structural equation modelling techniques were used to test a model of achievement related behaviours (effort) in mathematics based on support from social agents, competence beliefs, utility value and achievement goals. Several self-reported scales were administered. RESULTS: Results indicate that effort in mathematics is mainly explained by mastery goals and competence beliefs. As for the role of social agents, results demonstrated that the perception of parental support chiefly explained variables associated with the valuing of mathematics while teachers' support acted most on competence beliefs. CONCLUSIONS: Two main conclusions stem from our results. First, mastery goals have an important and significant impact on students' effort in the learning of mathematics. Second, the nature and the strength of the relationships between competence beliefs, utility value, achievement goals and effort are not significantly influenced by age and gender, at least in mathematics. PMID- 17908374 TI - Learning and memory of factual content from narrative and expository text. AB - BACKGROUND: Research on the presentation of information in narrative versus expository text genres is inconclusive with respect to the question of which is more beneficial for student learning. AIMS: We examine the effect of presenting factual content in either narrative or expository genres on student learning. We also consider relevant prior knowledge and working memory capacity (WMC) as potential mediating variables. SAMPLE: Ninety university undergraduate students. METHODS: Subjects studied circulatory system content embedded in either narrative or expository texts. Prior circulatory system knowledge, knowledge improvement (learning) and free recall were assessed. RESULTS: Learning and recall did not differ as a function of text genre overall, but did interact with prior knowledge. Learning from the narrative and one expository text was optimal at intermediate levels of prior knowledge, with higher knowledge readers benefiting more from the expository text compared with the narrative text. Prior knowledge was positively related to recall for the expository texts, but unrelated for the narrative text. Subjects' WMC did not predict learning or recall. CONCLUSIONS: Results suggest that narrative and expository processing differ with respect to integration of text content with prior knowledge. PMID- 17908373 TI - Integrated pictorial mnemonics and stimulus fading: Teaching kindergartners letter sounds. AB - BACKGROUND: The conclusion from a vast literature on literacy acquisition is that letter knowledge is one of the best predictors of literacy development. The question of the best way to teach children letter sounds has not, as yet, been answered satisfactorily. AIMS: The aim of this study was the evaluation of a computer training program using integrated-picture mnemonics combined with a fading procedure to teach children letter sounds. SAMPLE: Thirty-nine kindergartners attending mainstream primary education participated in this study. METHOD: A within-subject design was used. Each kindergartner learned letters under three conditions: (a) a fading condition in which letters are taught using a picture-supported first-sound mnemonics procedure in combination with a fading procedure; (b) an embedded condition in which letters are taught using the picture-supported first-sound-mnemonics procedure only and (c) a without-picture condition in which letters are taught using a first-sound procedure without picture support. Dependent measures included a productive and receptive letter sound test, and a first-sound isolation task. RESULTS: Productive letter-sound knowledge in the fading condition was better than in the other two conditions. In addition, kindergartners with good and those with poor first-sound isolation ability performed equally well in the fading condition. However, in the embedded and in the without-picture conditions, the kindergartners with good first-sound isolation ability outperformed those with poor isolation ability. CONCLUSION: These findings indicate that an integrated-picture mnemonics procedure combined with a fading procedure is effective in teaching kindergartners letter sounds and that the success of such a procedure does not depend on their initial first-sound isolation ability. PMID- 17908375 TI - The effects of adapting a writing course to students' writing strategies. AB - BACKGROUND: When writing a text, students are required to do several things simultaneously. They have to plan, translate and review, which involve demanding cognitive processes. In order to handle this complexity, writers need to develop a writing strategy. The two most well-defined writing strategies that have been identified, are those of a planning strategy and a revising strategy. AIMS: To establish whether students will be more competent in managing the complexity of writing when writing instruction is adapted to their habitual writing strategy, thus resulting in better texts. SAMPLE: 113 high school students (10th grade). METHOD: Students were randomly assigned to either the planning or the revising condition. To identify writing strategies, students completed a questionnaire concerning their planning and revising tendencies. To measure the level of writing skill, students' texts written during pre-test and post-test were analysed. RESULTS: The effect of instruction based on a planning strategy interacted with the level of planning or revising strategy: the greater the use of such a strategy, the larger the effect on writing skill. In contrast, the effect of instruction based on a revising writing strategy did not interact with the level of planning or revising strategy. Results imply that students with strong tendencies to plan or revise profited from writing instruction based on a planning strategy, while students with a low tendency to plan or revise profited more from instruction based on a revising strategy. CONCLUSION: Adapting writing instruction to students' level of writing strategy, is an effective approach for learning to write. PMID- 17908377 TI - Predicting academic self-handicapping in different age groups: the role of personal achievement goals and social goals. AB - BACKGROUND: Academic self-handicapping refers to the use of impediments to successful performance on academic tasks. Previous studies have shown that it is related to personal achievement goals. A performance goal orientation is a positive predictor of self-handicapping, whereas a task goal orientation is unrelated to self-handicapping. AIMS: The aim of this study was to examine the relationship between academic self-handicapping, goal orientations (task, performance-approach, performance-avoidance), social goals, future consequences and achievement in mathematics. An additional aim was to investigate grade-level and gender differences in relation to academic self-handicapping. SAMPLE: Participants were 702 upper elementary, junior and senior high school students with approximately equal numbers of girls and boys. RESULTS: There were no grade level or gender differences as regards the use of self-handicapping. The correlations among the variables revealed that, when the whole sample was considered, self-handicapping was positively related to performance goal orientations and pleasing significant others and negatively to achievement in mathematics. The results of hierarchical regression analysis showed that, in upper elementary and junior high schools, the association between achievement in mathematics and self-handicapping was mediated by performance-avoidance goals. In senior high school, only task goal orientation was a negative predictor of self handicapping. PMID- 17908376 TI - Test anxiety in UK schoolchildren: prevalence and demographic patterns. AB - BACKGROUND: Despite a large body of international literature concerning the antecedents, correlates of and treatments for test anxiety, there has been little research until recently using samples of students drawn from the UK. There is a need to establish some basic normative data for test anxiety scores in this population of students, in order to establish whether international research findings may generalize to UK schoolchildren. AIM: To collect some exploratory data regarding test anxiety scores in a sample of UK schoolchildren, along with socio-demographic variables identified in the existing literature as theoretically significant sources of individual and group differences in test anxiety scores. SAMPLE: Key Stage 4 students (1348): 690 students in the Year 10 cohort and 658 students in the Year 11 cohort, drawn from seven secondary schools in the North of the UK. METHOD: Data on test anxiety were collected using a self report questionnaire, the Test Anxiety Inventory (Spielberger, 1980) and additional demographic variables through the Student Profile Questionnaire. The factor structure of the Test Anxiety Inventory was explored using principal components analysis and multiple regression analysis used to predict variance in self-reported test anxiety scores from individual and group variables. RESULTS: The principal components analysis extracted two factors, worry and emotionality, in line with theoretical predictions. Gender, ethnic and socio-economic background were identified as significant predictors of variance in test anxiety scores in this dataset. Whether English was an additional, or native, language of students did not predict variance in test anxiety scores and year group was identified as a predictor of emotionality scores only. CONCLUSION: Variance in the test anxiety scores of Key Stage 4 students can be predicted from a number of socio-demographic variables. Further research is now required to assess the implications for assessment performance, examination arrangements and appropriateness of using a North American measure of test anxiety in a UK context. PMID- 17908378 TI - The effects of selective schooling and self-concept on adolescents' academic aspiration: an examination of Dweck's self-theory. AB - BACKGROUND: Dweck has emphasized the role of pupils' implicit theories about intellectual ability in explaining variations in their engagement, persistence and achievement. She has also highlighted the role of confidence in one's intelligence as a factor influencing educational attainment. AIM: The aim of this paper is to develop a model of achievement aspiration in adolescence and to compare young people who are educated at a selective grammar school with those who attend a non-selective 'secondary modern' school. SAMPLE: The sample consisted of 856 English secondary school pupils in years 7 and 10 from two selective and two non-selective secondary schools. METHOD: Questionnaires were completed in schools. RESULTS: The findings are consistent with the model, showing that achievement aspiration is predicted directly by gender, school type and type of intelligence theory. Importantly, school type also affects aspirations indirectly, with effects being mediated by confidence in one's own intelligence and perceived academic performance. Intelligence theory also affects aspirations indirectly with effects being mediated by perceived academic performance, confidence and self-esteem. Additionally, intelligence theory has a stronger effect on aspirations in the selective schools than in the non-selective schools. CONCLUSIONS: The findings provide substantial support for Dweck's self theory, showing that implicit theories are related to aspirations. However, the way in which theory of intelligence relates to age and gender suggests there may be important cross-cultural or contextual differences not addressed by Dweck's theory. Further research should also investigate the causal paths between aspirations, implicit theories of intelligence and the impact of school selection. PMID- 17908379 TI - Understanding teachers' perceptions of the motor difficulties of children with developmental coordination disorder (DCD). AB - BACKGROUND: Children with developmental coordination disorder (DCD) are often identified by classroom teachers and the identification process relies heavily on teachers' perceptions. The literature would suggest that teachers' perceptions may be influenced by a child's gender, behaviour and the type of motor problem they demonstrate. To date, the influence of these factors on teachers' perceptions of children with DCD has not been empirically tested. AIM: This study investigated whether child gender, behaviour and type of motor problem influenced teachers' ratings of concern and importance of intervening for children with motor difficulties. SAMPLE: One hundred and forty-seven teachers of children from 6 to 9 years of age participated in this study. METHOD: Hypothetical case scenarios were developed that experimentally manipulated the factors of child gender (male/female), behaviour (disruptive/non-disruptive) and type of motor problem (fine motor/gross motor). Teachers were given two case scenarios of the same gender (that varied by behaviour) and rated: (a) their degree of concern about children's motor problems and (b) how important they thought it was for the child to receive intervention for that problem. RESULTS: The effect of child gender on teachers' perceptions depends upon the type of motor problem. While child behaviour had a marginal influence on teachers' perceptions, interestingly, teachers appeared to recognize motor problems only in the absence of disruptive behaviour. The type of motor problem demonstrated also influenced teachers' perceptions. CONCLUSION: This study provides preliminary insight into factors that influence teachers' perceptions of children with DCD with clear implications for the classroom identification of children with DCD. PMID- 17908380 TI - Teacher responses to bullying in relation to moral orientation and seriousness of bullying. AB - BACKGROUND: Little research has focused on factors influencing teachers' decisions about whether and how to intervene in bullying incidents. Such factors have the potential to influence the role of teachers as agents in counteracting bullying. AIMS: To examine: (a) whether moral orientation predicts teachers' responses to bullying, (b) the role of perceived seriousness of an incident in moderating responses to bullying and (c) factors that are important to teachers when deciding whether to intervene. SAMPLE: Primary, middle and high school teachers (N=127) were recruited during staff meetings at five schools. METHODS: Moral orientation was measured using a modified version of Caputo's (2000) Sanctioning Voice Index (SVI); other questionnaires were specifically designed for this study. Correlational and hierarchical multiple regression analyses examining how moral orientation and seriousness predict teachers' responses to bullying were performed. RESULTS: As anticipated, care moral orientation predicted a problem-solving response, while justice orientation predicted a rules sanctions response. Care and justice orientations also interacted to predict rules-sanctions, but not problem-solving responses. However, seriousness of an incident accounted for the majority of variance (46% for rules-sanctions and 40% for problem-solving responses). Seriousness did not moderate the relationship between moral orientation and responses to bullying. CONCLUSIONS: While teachers' moral orientation does impact upon the kinds of responses to bullying they choose, seriousness of the incident is more important. However, seriousness as perceived by teachers may not be consistent with impact on students. Implications for teacher education and policy are discussed. PMID- 17908381 TI - Developing identities and attitudes in musicians and classroom music teachers. AB - BACKGROUND: The Western classical training of many secondary music specialist teachers may be inappropriate for the demands of the contemporary secondary school classroom, leading to a conflict between their self-concepts as 'musicians' and as 'teachers'. AIMS: To undertake a short-term longitudinal comparison of the developing identities and the attitudes of a group of intending specialist secondary music teachers, during the transition into their first teaching post, with a group of music students from university and conservatory backgrounds. SAMPLE: Twenty-nine trainee music teachers completed Phases 1 and 2 of the study during their final weeks of training and during the second term of their teaching career, and a comparison group of 29 final-year undergraduate music students did so in the first and last terms of their final year. METHOD: A specially devised composite Musical Careers Questionnaire gathered information in both phases about self-efficacy in music and in teaching, identification with professional groups in these two domains, and attitudes towards important skills for musicians and teachers. RESULTS: A series of ANOVAs comparing the student groups' scores in each of the two phases revealed no significant main effects or interactions for either of the self-efficacy measures, a significant group effect for professional group identification, and some changes in the attitude measures. CONCLUSION: Although participants' views of their own general effectiveness as teachers and as musicians changed very little over the period of the study, their attitudes towards music teaching and perceptions of the skills required showed some changes. PMID- 17908382 TI - Future time orientation predicts academic engagement among first-year university students. AB - BACKGROUND: Enhancing student engagement is considered an important strategy for improving retention. Students' Time Perspective is an under-researched factor that may significantly influence student engagement. AIMS: This study examines interrelationships between elements of student engagement and relationship with Time Perspective. We propose that there are significant relationships between psychological and behavioural elements of student engagement. We also posit that time orientation is an important factor in facilitating psychological and behavioural elements of student engagement. SAMPLE: Participants (N=347) were first-year undergraduate students who had completed one semester of study and re enrolled for a further semester of study at an Australian university. METHODS: Participants were surveyed using instruments designed to measure Academic Application, Academic Orientation (McInnis, James, & Hartley, 2000), Time Perspective (Zimbardo & Boyd, 1999), the shortened version of the Study Process Questionnaire (Fox, McManus, & Winder, 2001) and hours spent preparing for class. RESULTS: There were interrelationships between the elements of student engagement (e.g. Academic Application) with productive educational behaviours (e.g. deep approach to learning). Students' perceptions of time appeared as a key factor mediating levels of Academic Application and Academic Orientation. Orientation to the Future emerged as a significant predictor of these elements of engagement. CONCLUSIONS: Future orientation emerged as an important factor mediating students' academic engagement in these students who completed one semester of study. Interventions focusing on the development of time perspective may be helpful in encouraging and supporting academic engagement and, ultimately, persistence in higher education. PMID- 17908383 TI - The revised learning process questionnaire: a validation of a Western model of students' study approaches to the South Pacific context using confirmatory factor analysis. AB - BACKGROUND: Research evidence seems to suggest that the social and cultural environments influence students' approaches to their study. This social and cultural contention has led to the rethinking and reconceptualization of theories (e.g. Biggs, 1987; Marton & Saljo, 1976) pertaining to student approaches to learning (SAL) in academic settings. AIMS: The present research discusses two separate empirical studies on student learning approaches situated in the South Pacific region with two respective cohorts of secondary students. Study I involved the examination of secondary Pacific Islands students in their learning approaches using a modified version of Biggs' (1987) original Learning Process Questionnaire (LPQ). Study II involved the administration of a revised version of the LPQ (R-LPQ-2F; Kember, Biggs, & Leung, 2004) to another cohort of secondary Pacific Islands students. SAMPLE: The first sample included 2,150 (1,285 girls, 865 boys) students and the second sample included 2,295 (1,363 girls, 932 boys) students. METHODS: The factor structures of approaches to learning were examined by means of confirmatory factor analysis (CFA) using the LISREL program. Different a priori models were hypothesized and tested. RESULTS: Results of Study I indicated a two-factor structure solution to Biggs' LPQ, supporting Richardson's (1994) theoretical model of learning and emphasized the factors of Reproducing and Meaning. Study II indicated a hierarchical organization of two main study approaches - deep and surface - that are structured as higher-order factors and a defined by four first-order factors. CONCLUSION: The results from the two studies accentuate the important argument for the rethinking and reconceptualization of learning approaches, as well as for the redevelopment and modification of learning inventories such as the LPQ. They also suggest the importance of situating the theoretical paradigm of learning approaches in a social and cultural environment. PMID- 17908394 TI - Multicentre trial of preimplantation genetic screening reported in the New England Journal of Medicine: an in-depth look at the findings. AB - A randomized clinical trial of 406 patients with advanced maternal age by Mastenbroek and co-workers recently published in the New England Journal of Medicine showed a significant decrease in pregnancy outcome after preimplantation genetic screening (PGS). It is our opinion that this study suffers from a number of insurmountable inaccuracies and that these are either a direct consequence of the inexperience of the team or of a general disregard of vital guidelines reported in the literature. Most importantly, the authors show that in their hands embryo biopsy may affect as many as half the embryos. The error rate was not presented, shedding doubt on the authors' abilities to reliably diagnose the biopsied cells. An evaluation of the study indicates that poor biopsy technique, sub standard fixation and FISH methods, poor IVF outcomes and inappropriate patient selection are the cause of the discouraging results obtained by these authors rather than problems inherent to PGS. PMID- 17908396 TI - Tubal disease: towards a classification. AB - Tubal disease is a major cause of infertility. The amount of damage can vary greatly in extent, anatomical location and nature. For women with infertility due to tubal disease, prognostication for pregnancy often remains unclear and there is no universally accepted classification. A classification system that reliably distinguishes infertile patients with tubal disease into favourable and unfavourable groups would be useful if subsequent management could depend on this assessment, especially if the classification is able to define which group of patients would benefit most from interventions such as surgery. The progress of IVF questions the contribution of the Fallopian tube to the successful achievement of pregnancy in infertile women. Nonetheless, several studies reveal that severity is the key factor in the determining outcome, and the classifications reviewed in this paper imply that women with tubal disease could be categorized into prognostic groups using a simple classification system based on severity. However, prospective trials are needed to validate and assert the usefulness of any particular classification. PMID- 17908397 TI - Tubal assessment tests: still have not found what we are looking for. AB - Interest in tubal assessment is as old as interest in fertility and infertility. The Fallopian tube is a particularly complex structure and, as such, an ideal method for its clinical assessment is very difficult to obtain. As a result, a number of different methods have been suggested. Some of these methods are more complementary to each other rather than potential substitutes for one another. Some have been used for many years with a clear evidence base for their performance as diagnostic tests. For other, relatively new tests, very little evidence about their performance is available. Research is moving from a purely anatomical approach (are the tubes open or blocked?) to encompassing functional enquiry (are the open tubes functional and, if not, are there interventions with which fertility performance can be improved?). The available evidence, or lack thereof, for the most commonly used tubal assessment tests is reviewed in this paper. Many questions remain, which, despite the increasing success of IVF, will continue to challenge and stimulate specialists and the public, who are interested in ways to maximize spontaneous as opposed to assisted fertility. PMID- 17908398 TI - Proximal tubal disease: the place for tubal cannulation. AB - Tubal disease is the cause of subfertility in approximately 30% of women, and 10 25% of these are due to proximal tubal obstruction. False-positive diagnosis of proximal tubal obstruction can be as high as 50%. A decrease in expertise in tubal microsurgery has resulted largely from the use of IVF as the treatment option for most causes of infertility and more specifically for tubal factor infertility. Selective salpingography and tubal cannulation have a unique role in the management of tubal infertility and should be offered to selected candidates prior to IVF. Tubal cannulation can be used effectively to restore patency in a proportion of cases of proximal tubal obstruction thus avoiding the need for expensive assisted reproductive techniques. This review examines the evidence supporting the effectiveness of tubal cannulation and aims to enhance awareness of the procedure as an option for the management of female subfertility secondary to isolated proximal tubal obstruction. PMID- 17908399 TI - Place of transvaginal fertiloscopy in the management of tubal factor disease. AB - In the case of tubal disease, selection of patients is critical to decide whether tubal surgery or IVF should be proposed as the therapeutic option. Selection is based on tubal lesions, including the aspect of tubal mucosa and tuboperitoneal environment. Therefore, non-invasive diagnostic methods have proved to be insufficient, and endoscopy is mandatory. Fertiloscopy represents an attractive alternative. Fertiloscopy is minimally invasive, reproducible, and at least as precise as lap and dye. Moreover, fertiloscopy and not laparoscopy allows the routine practice of salpingoscopy and microsalpingoscopy. On this basis, fertiloscopy should be adopted as the standard procedure for tubal evaluation in the management of tubal disease. PMID- 17908400 TI - Identical triplets born in Britain. PMID- 17908401 TI - Controversies in the management of ectopic pregnancy. AB - Ectopic pregnancy is a common clinical problem, but there appears to be much controversy surrounding the surgical management of its occurence. This paper reviews the available evidence on the management of ectopic pregnancy. The discussion focuses initially around the choice of medical versus surgical treatment. Next, the question is addressed that if surgical management is deemed necessary, whether the approach should be laparoscopic or via open laparotomy. Lastly, if surgery is undertaken, should salpingectomy or salpingotomy be performed? Laparoscopy will remain the main method of treatment for women with ectopic pregnancy, as it provides obvious advantages over open surgery. On balance, salpingotomy should be the surgical treatment of choice for the majority of women with ectopic pregnancy, as it results in a higher subsequent pregnancy rate, although there is a slightly higher recurrent ectopic pregnancy rate and persistent trophoblastic disease rate when compared with women treated with salpingectomy. There is also a place for medical treatment of women with low concentrations of human chorionic gonadotrophin. A variable dosing methotrexate regimen is more effective compared with single dose regimen, and the fixed multiple regimen is associated with a high rate of side effects. PMID- 17908402 TI - Reversal of tubal sterilization versus IVF in the era of assisted reproductive technology: a clinical dilemma. AB - Two treatment options are available to women who wish to become pregnant after having had tubal sterilization: microsurgical reversal or IVF. The first approach is designed to restore tubal function, whereas the second replaces it. The first, to be successful, requires the presence of sufficient tubal length and normal or treatable fertility parameters. Treatment should therefore be individualized, based upon the findings of the couple's investigation, their wishes and the costs involved. The age of the female is the most important factor that affects the outcome with both treatment options. The live birth rate per cycle with IVF is 28%, but only 65.8% are singletons; 31.0% are twins and 3.2% triplets or more. Microsurgical tubal anastomosis yields a birth rate that exceeds 55%, without increased risk of multiple pregnancy. It offers the couple multiple cycles in which to achieve conception naturally, and the opportunity to have more than one pregnancy from a single intervention. The real dilemma lies with the 'industrialization' of IVF, and its frequent use as primary treatment for infertility. The dilemma is heightened by the fact that reconstructive tubal microsurgery is being taught and practised less and less, thereby eliminating this credible surgical option in most centres. PMID- 17908403 TI - Management of severe early ovarian hyperstimulation syndrome by re-initiation of GnRH antagonist. AB - Several approaches have been proposed for the management of OHSS that reduce, but do not completely eliminate the incidence of human chorionic gonadotrophin (HCG) induced early severe OHSS. Three women diagnosed with polycystic ovarian syndrome underwent ovarian stimulation for IVF using a gonadotrophin-releasing hormone (GnRH) antagonist protocol. Three days after oocyte retrieval, severe early OHSS was diagnosed by analysis of haematocrit (Ht), white blood cell (WBC) count, serum urea, and ultrasonographic assessment of ovarian size and ascitic fluid. On the same day, antagonist administration was re-initiated and continued daily for a week, while all embryos were cryopreserved. No progression of severe early OHSS was observed in any of the patients. A marked decrease of Ht, WBC count, ovarian volume and ascitic fluid was observed during 1 week of follow-up, and none of the patients required hospitalization. GnRH antagonist re-initiation might represent a new strategy for flexible management of patients with established severe early OHSS. Based on the flexibility of the approach, if severe OHSS does not occur, patients may proceed to embryo transfer, while if severe early OHSS ensues, antagonist administration combined with embryo cryopreservation appear to be associated with prevention of life-threatening OHSS, facilitation of regression of severe OHSS to a moderate form and avoidance of patient hospitalization. PMID- 17908404 TI - Maternal serum hormone concentrations for prediction of adverse outcome in threatened miscarriage. AB - Many serum markers have been investigated in attempts to predict the outcome of pregnancy in the first trimester, with varying degrees of success. The objective of this study was to investigate whether they can be related to pregnancy outcome in women presenting with first trimester threatened miscarriage. A cohort study of women attending the Early Pregnancy Unit of a London teaching hospital was studied. A total of 122 women presenting with bleeding in the first trimester and an ongoing pregnancy, and 33 women undergoing termination of pregnancy, were recruited. The main outcome measures were gestation at delivery, birth weight and the incidence of adverse pregnancy outcome. Inhibin A, activin A, human chorionic gonadotrophin (HCG), pregnancy-associated plasma protein-A and follistatin concentrations were all significantly lower in women who subsequently miscarried when compared with live births. Serum HCG concentrations were significantly higher in cases of threatened miscarriage compared with controls (P = 0.0009). Logistic regression analysis indicated that inhibin A alone provided the best predictor for first trimester miscarriage. This pilot study suggests that placental hormone concentrations could be useful in predicting adverse pregnancy outcome in women presenting with threatened miscarriage. Inhibin A was best at predicting the likelihood of subsequent miscarriage in this group. PMID- 17908405 TI - Patients' preferences for intrauterine insemination or in-vitro fertilization. AB - Patients' preferences for intrauterine insemination (IUI) relative to IVF were assessed using trade-off interviews, and the number of IUI cycles they would undergo before changing to IVF. A total of 73 couples undergoing IUI with a total of 111 interviews were included. Scenarios were offered where pregnancy chance after IUI was varied against a fixed pregnancy rate after IVF. The impact of multiple pregnancy risk on the couple's preference was also investigated. Interviews were held before starting IUI, after three or four IUI cycles and after six IUI cycles. With decreasing probability of ongoing pregnancy after IUI, an increasing number of couples switched their preference from IUI to IVF. This switch occurred after six cycles at a significantly higher (P = 0.01) mean cumulative pregnancy rate (53%) compared with other groups (31%). With increasing risk of multiple pregnancy, preference for IUI declined only slightly, with mean risks of 73, 78 and 83% of a multiple pregnancy for the three groups respectively. In conclusion, at baseline and after three cycles of IUI the majority of couples undergoing IUI preferred continuation of IUI over IVF. A clear shift in preference towards IVF occurred after six cycles. Risk of multiple pregnancy did not affect preference for IUI with ovarian stimulation. PMID- 17908406 TI - Five years of single embryo transfer with anonymous and non-anonymous oocyte donation. AB - Single embryo transfer (SET) has been the main embryo transfer strategy in the oocyte donation programme at the authors' clinic since 2000. The primary aim of this study was to evaluate the effect of SET on the clinical outcome in an unselected group of oocyte recipients. A retrospective analysis of the outcome in 142 recipient cycles (116 from anonymous donors; 26 from known donors) was performed. The oocytes from each anonymous donor were shared between two recipients if at least 10 oocytes were obtained. The proportion of SET of all fresh transfers was 77.3%. The clinical pregnancy rate (CPR) was 43.2% and the delivery rate 31.1% per embryo transfer. The outcome was similar in recipients undergoing anonymous and non-anonymous donation. The delivery rates were similar after SET (30.4%) or double embryo transfer (DET) (33.3%), whereas the twin rate was 0% after SET and 40% after DET. The implantation rate was significantly better (P < 0.01) with good-quality embryos (54.7%) compared with non-optimal embryos (27.1%). Of 152 frozen-thawed embryo transfer cycles, 78.9% were SET. The CPR was 28.3% and the twin rate was 7.1%. In fresh oocyte donation cycles, elective SET can be recommended if the embryo quality is considered good, and always if there is a contraindication for twin pregnancy. PMID- 17908407 TI - Semen quality in a population of volunteers from the province of Barcelona. AB - A prospective study was carried out on 1005 male volunteers residing in the province of Barcelona. Participants were asked to complete a questionnaire concerning their health status and lifestyle, and provided a semen sample for analysis. Sperm concentration and percentages of motile, and normal forms were evaluated. The effect of smoking, alcohol consumption, stress and previous disease on semen parameters was also evaluated. Normal parameters were found in 22% of volunteers and sperm counts and/or percentage motility below normal threshold values were found in 78%. Asthenozoospermia detected in 62% and oligozoospermia in 17% of volunteers. No statistically significant association was found between semen quality and age, with the exception of a decrease in semen volume (P = 0.04) and progressive motility (P = 0.01). No statistically significant differences in semen parameters were found between smokers and non smokers or between males who consumed alcohol versus those that did not. However, stress had a negative effect on sperm concentration. In conclusion, the prevalence of normal semen parameters in the study population evaluated was markedly lower than that reported for the general population (60-70%). Since sperm concentration did not appear to decrease with age, the decrease observed in the last decades may be related to exposure to environmental toxicants during the fetal period, as previously suggested. PMID- 17908408 TI - Timing intra-Fallopian transfer procedures. AB - With the gradual decline in the use of zygote intra-Fallopian transfer (ZIFT), current practice is to offer ZIFT almost exclusively to patients with repeated implantation failure (RIF). For practical reasons, the procedure is sometimes deferred by 1 day and embryo intra-Fallopian transfer (EIFT) is performed. The aim of the present study was to compare the reproductive outcome of ZIFT versus EIFT. In a retrospective analysis, 176 patients who failed in 7.65 +/- 3.7 previous IVF cycles underwent 200 ZIFT and 73 EIFT procedures. Implantation and live birth rates were compared for both groups. Patients in both groups were found comparable for demographic and clinical parameters. Similar numbers of oocytes were retrieved and fertilized in both groups, and 5.2 +/- 1.2 zygotes/embryos were transferred. Implantation and live birth rates (10.5 and 26.5% versus 10.9 and 24.7% for ZIFT and EIFT respectively) were comparable. It is concluded that tubal transfer of zygotes and day-2 cleavage stage embryos are equally effective. PMID- 17908409 TI - European Research Council issues research grants. PMID- 17908410 TI - Emerging technologies for the molecular study of infertility, and potential clinical applications. AB - The techniques currently used to treat infertility cases are quite limited in their capabilities, due to an incomplete understanding of the molecular activities of germ cells. Fortunately, several technologies are presently being researched that should aid the understanding of the various molecular causes of germ cell pathologies. This review discusses microarray technology, proteomics, metabolic profiling, the PolScope, atomic force microscopy and microfluidics. These technologies have all seen success in preliminary studies, and promise directly or indirectly to improve the low success rates of IVF and other related therapies. However, their widespread use in laboratories and clinics may not be seen until preliminary studies confirming their safety and effectiveness are published, and until standardized protocols for their utilization are established. PMID- 17908411 TI - Unilateral ovarian drilling in polycystic ovarian syndrome: a prospective randomized study. AB - Ovarian drilling is a well-accepted intervention for ovulation induction in clomiphene citrate-resistant polycystic ovarian syndrome (PCOS) patients. The aim of this study was to evaluate the effect of unilateral and bilateral ovarian drilling by electrocautery in PCOS women. In this prospective randomized clinical study, 87 patients with ovulation failure as a result of PCOS were randomly allocated to either unilateral (group A; n = 43 patients) or bilateral (group B; n = 44 patients) laparoscopic ovarian drilling by electrocautery. The average time required for unilateral ovarian drilling was shorter than for bilateral drilling. In patients who ovulated after drilling, there was a significant fall in serum LH concentration (group A, P < 0.05, group B, P < 0.05). Ovulation, pregnancy and miscarriage rates were similar in both groups. It seems that unilateral ovarian drilling in PCOS is effective, less time-consuming and probably associated with fewer complications. PMID- 17908412 TI - Prevalence of chromosomal aberrations in Mexican women with primary amenorrhoea. AB - Primary amenorrhoea refers to the absence of menarche by the age of 16-18 years in the presence of secondary sexual characteristics, and occurs in 1-3% of women of reproductive age. To study the prevalence of chromosomal abnormalities and the different options available for clinical management of women in Mexico with primary amenorrhoea, a cross-sectional study was conducted in 187 women with primary amenorrhoea referred from Department of Reproductive Medicine of Morones Prieto Hospital, IMSS in Monterrey, Mexico during 1995-2003. Peripheral blood lymphocytes were cultured for chromosomal studies by the standard methods. Numerical or structural abnormalities of the sex chromosome were found in 78 women (41.71%). These women were classified into four categories: X-chromosome aneuploidies (22.99%: 12.83% pure line and 10.16% mosaicism association with a 45, X cell line); presence of chromosome Y (10.70%); structural anomalies of the X chromosome (4.28%); and marker chromosomes (3.74%). In conclusion, the prevalence of chromosomal abnormalities in Mexican women with primary amenorrhoea is within the range (24-46%) reported in world literature. Chromosomal analysis is absolutely necessary for appropriate clinical management of these patients. PMID- 17908413 TI - Understanding the role of LH: myths and facts. AB - This review summarizes a series of lectures given at a recent Continuing Medical Education meeting in Hamburg, Germany (May 2007), aiming to understand the role of luteinizing hormone (LH) in follicular development during the natural menstrual cycle and controlled ovarian stimulation. Clinical situations and target groups of patients who might benefit from LH supplementation during their ovarian stimulation were discussed and defined. The lectures updated knowledge on the physiology of LH during the normal menstrual cycle and the role of LH in ovarian stimulation. The concept of the 'LH window' was presented, and the use of LH supplementation in different groups of patients undergoing controlled ovarian stimulation was discussed, including those with advanced age, hypogonadotrophic hypogonadism, pituitary down-regulation and poor response. In addition, the different ways of using LH or human chorionic gonadotrophin supplementation in ovulation induction protocols were described. PMID- 17908417 TI - One dental hygienist's role in clinical research. PMID- 17908418 TI - Advanced degree seeking students' satisfaction with online courses at UMKC--an early investigation. AB - The use of online instruction is becoming more prevalent and transcends many disciplines. Nursing has been at the forefront of health professions utilizing distance education. Nearly half of all predoctoral dental programs report use of web-based or distance delivery. Comparatively, 22% of dental hygiene programs report use of this somewhat new approach to teaching. Distance learning provides a means for increasing access to and enrollment in dental hygiene programs. Individuals who cannot physically attend courses in dental hygiene benefit from the institutions that offer web-based classes. In today's environment, more individuals seek to advance their educational needs. Advanced dental hygiene degree programs may benefit by providing distance learning. Since 2000, the University of Missouri - Kansas City (UMKC), Division of Dental Hygiene has offered its degree completion program online. Its master's degree online program has been offered since 2001. PURPOSE: The purpose of this study is to describe the pre- and post-course graduate and degree completion students' perceptions of web-based courses offered at UMKC. METHOD: Over the course of 3 years, all students enrolled in required fall semester online classes were selected and agreed to participate in the study. Students were asked to fill out questionnaires prior to and at the completion of required online courses. There were no identifiers on the questionnaires. The surveys used in the study were derived from an instrument used by Wills and Stommel who examined graduate nursing students' perceptions of web-based courses. RESULTS: Results indicate that students enjoy this method of learning. All students reported they would enroll in another online course if given the opportunity. CONCLUSION: Based on favorable student perceptions, UMKC will continue to provide this method of learning to students seeking advanced degrees in dental hygiene. PMID- 17908419 TI - Validation of the National Board Dental Hygiene Examination. AB - The National Board Dental Hygiene Examination program is a part of the process for licensing dental hygienists. The examination assesses theoretical and applied knowledge in the basic biomedical, dental, and dental hygiene sciences, as well as community health. Standards for licensure examinations recommend that test publishers demonstrate a relationship between examination content and actual practice. METHOD: To this end, a validity study was conducted, which involved the definition of the domain of entry-level dental hygiene practice using 56 competencies; the conduct of a practice analysis survey designed to rate the importance of these competencies; and the linking of competencies to content elements in accordance with the competencies' importance ratings. Of the 3941 surveys distributed, 1841 participants responded and, of these, 1284 were full time practitioners. The importance ratings for the competencies were translated into numbers of items. The number of items devoted to each competency was distributed across all applicable elements of the existing content specifications based upon the knowledge needed to support the realization of the competency. RESULTS AND DISCUSSION: The findings confirmed the adequacy of the content specifications in effect prior to 2005. However, based on this validity study, 2 sub areas of relatively little significance were eliminated, and 2 new areas were introduced. Specifically, Clinical Testing under Assessing Patient Characteristics (one item) and Professional Methods of Administering Fluorides under Using Preventive Agents (one item) were eliminated, and Dental Hygiene Treatment Strategies was incorporated with 4 items, and Professional Responsibilities was added with a total of 28 items. PMID- 17908420 TI - Service-learning and dental hygiene: a literature review. AB - Dental hygienists should be aware of the concept of service-learning and how it can enhance dental hygiene education. A look at national documents and other relevant historical literature will be reviewed, in addition to more recent books and articles to define service-learning. Although perceived as a new teaching strategy, service-learning has its roots in experiential education. Several definitions of service-learning have emerged, which take into consideration the reciprocal needs of the student and community, bridging academic theory with community service, and instilling civic responsibility through a reflective component. Considering the advantages and disadvantages, service-learning seems to be a good methodology for dental hygiene education. Recommendations include the expansion of the ADA Accreditation Standards to include service-learning, and incorporation of an operational definition of community-based oral health programs, as well as the term service-learning in its Definition of Terms. PMID- 17908421 TI - Oral assessment of children with an autism spectrum disorder. AB - PURPOSE: The study assessed the oral health status of children with an autism spectrum disorder (ASD) to help establish the oral health needs of this population. METHODS: Oral assessments were conducted on 39 children with an ASD and 16 children with other developmental disabilities (DD), solicited from 3 different schools. Conditions assessed were bacterial plaque, gingivitis, dental caries, restorations, bruxism, delayed eruption/missing teeth, oral infection, developmental anomalies, injuries, occlusion, salivary flow, and oral defensiveness. RESULTS: Chi-square and Fisher's exact test of significance were used to compare groups. Young children with an ASD who resided with parents showed significantly more signs of bruxism than the comparison groups. Likewise, older children who lived at the residential school manifested significantly more gingivitis. No other significant differences existed when age and residence were considered for children with an ASD. When comparing children with ASD to those with another DD, the latter group showed significantly more oral injuries, abnormal salivary flow, and developmental anomalies. Children with an ASD displayed the following percentages for clinically visible conditions: plaque (85%), gingivitis (62%), and caries (21%). Approximately half of the children with ASD were orally defensive. CONCLUSIONS: Children with an ASD appear to have oral conditions that might increase the risk of developing dental disease. The extent of risk is unclear and needs further investigation. PMID- 17908422 TI - Articaine: a new alternative in dental hygiene pain control. AB - PURPOSE: Local anesthesia administration is integral to pain control in dental hygiene. As of 2006, 40 licensing jurisdictions in the United States include local anesthesia administration in the scope of dental hygiene practice. While the risks associated with use of intraoral local anesthesia are not great, careful attention to recommended practices helps foster patient safety. As new products are introduced, it is important to study their advantages and limitations to see where they fit into dental hygiene practice. An amide local anesthetic agent, articaine, that has been available in Europe for over 20 years, was approved for US distribution in 2000. METHODS: The purpose of this article is to review the current peer reviewed literature on the characteristics of articaine so it can be incorporated into dental hygiene practice when indicated. RESULTS: Rather than simply using one agent for all procedures, patient care is enhanced when local anesthetics are selected based on the unique needs of the procedure, the patient and with safety and effectiveness in mind. PMID- 17908423 TI - An examination of the bleeding complications associated with herbal supplements, antiplatelet and anticoagulant medications. AB - Dental professionals routinely treat patients taking prescription, nonprescription, and herbal medications that are known or have the potential to alter bleeding. Prescription anticoagulant and antiplatelet medications, as well as over-the-counter drugs such as aspirin, are typically taken to reduce the risk of thromboembolic events, including stroke. Herbal supplements are widely used for a variety of indications, and both patients and health care practitioners are often unaware of the anticoagulant and antiplatelet effects that occur as either predictable pharmacologic effects or adverse side effects of herbal medicines. In addition, patient use of these herbal supplements is usually undisclosed to health care providers. The purpose of this literature review is to examine the mechanisms of action of drugs and herbs that alter bleeding, and to educate dental professionals as to the proper care and management of patients using these medications. Decision-making strategies, including interpretation of laboratory tests, and when to discontinue the use of these medications are discussed. Patients undergoing routine dental and dental hygiene procedures do not need to discontinue the use of anticoagulant and antiplatelet medications. However, alterations in drug use may be required for those patients undergoing invasive surgical procedures. It is recommended that herbal supplements must be discontinued 2 weeks prior to receiving invasive surgical procedures. Dental practitioners must learn to weigh the risks of discontinuing drug therapy against the potential risks to patients, and implement risk reduction strategies to minimize adverse bleeding complications associated with dental treatment. PMID- 17908424 TI - [c-Jun NH2-terminal kinase and extracellular signal-regulated protein kinase signaling pathways in regulation of benzo(a)pyrene-induced c-Jun activation in human embryo lung fibroblasts]. AB - OBJECTIVE: To investigate the role of mitogen activated protein kinases (MAPKs) signaling pathways in the regulation of benzo(a)pyrene (B(a)P)-induced c-Jun activation in human embryo lung fibroblasts (HELFs). METHODS: HELFs were cultured with 2.0 micromol/L B(a)P for various time (0, 3, 6, 12, 24 h) or with various concentration of B(a)P (0.0, 0.5, 1.0, 2.0 micromol/L) for 12 h. Western blot was performed to examine the effect of B(a)P on c-Jun activation. The dominant negative mutants of p38, c-Jun NH2-terminal kinase (JNK) and extracellular signal regulated protein kinase (ERK) were applied to establish stable transfectant, and to detect the relationship of MAPK signal molecules and c-Jun activation in B (a) P-treated cells. RESULTS: B(a)P treatment resulted in a marked activation of c Jun in time-dependent manner with a peak at 12 h (the densitometric ratios of phosphorylated c-Jun Ser63, Ser73 to actin were 20.1, 15.2 times for control respectively) and in dose-dependent manner. However, there was no evident change on total c-Jun expression in B(a)P-treated HELFs. Moreover, B(a)P-induced activation of c-Jun was inhibited by stable expression of dominant negative mutants of JNK or ERK, but not by dominant negative mutant of p38. CONCLUSION: JNK and ERK signaling pathways, but not p38 pathway regulate B(a)P-induced c-Jun activation in HELFs. PMID- 17908425 TI - [Protective effects of vigabatrin and atropine against dimethoate induced intoxication in mice]. AB - OBJECTIVE: To investigate the protective effects of vigabatrin and atropine against the acute toxicity of dimethoate in male Kun-min mice. METHODS: The therapeutic schedules of vigabatrin (50 or 100 mg/kg) and (or) atropine (2.5 or 5.0 mg/kg) were performed according to the L(9) (3(4)) orthogonal test table. The survival time, the righting reflex and the onset of muscle fasciculations were observed after the administration of dimethoate. RESULTS: First, the main effects of vigabatrin, atropine and the interaction between them were statistically significant in the Univariate analysis of the survival time at the alpha level of 0.05 (F(V)= 4.73, P(V)= 0.015, F(A)= 50.88, P(A)= 0.000, F(VxA)= 4.17, P(VxA)= 0.007). The range of atropine was more than double of that of vigabatrin or their interaction (R(A)> 2RV or 2R(VxA)). So not only atropine and vigabatrin but also their combination interaction protected mice against dimethoate lethality. The atropine played the major role in diminishing the lethality induced by dimethoate. Second, only vigabatrin, while not atropine, delayed the mice from dimethoate-induced muscle fasciculation according its statistical results (F(V)= 6.87, P(V)= 0.003, F(A)= 0.03, P(A)= 0.968, F(VxA)= 1.134, P(VxA)= 0.356). It should be noted that vigabatrin could not completely prevent dimethoate induced muscle fasciculation in the test. At last, both atropine and vigabatrin could maintain the righting reflex in the intoxication, however there was no interaction between them (F(V)= 5.81, P(V)= 0.006, F(A)= 9.05, P(A)= 0.001, F(VxA)= 1.34, P(VxA)= 0.257). CONCLUSION: Combined treatment with atropine and vigabatrin in the organophosphates intoxication seems reasonable and acceptable. PMID- 17908426 TI - [Acute blended pesticides poisoning in 40 patients.]. PMID- 17908427 TI - [Relationship between heat shock protein 72 and DNA genetic damage in peripheral blood lymphocytes of coke oven workers]. AB - OBJECTIVE: To investigate the relationship between heat shock protein 72 (Hsp72) and DNA genetic damage in peripheral blood lymphocytes of coke oven workers and the role of Hsp72 in protection of cells from genetic damage induced by coke oven emissions. METHODS: Two hundred and sixty-seven coke oven workers and thirty controls without occupational PAHs exposure were investigated. Benzo[a]pyrene concentrations in the ambient air individually collected were assayed by high performance liquid chromatography (HPLC). Western Blot was used to measure Hsp72 levels and Comet assay was used to evaluate DNA damage degree. Personal information was collected by questionnaire. RESULTS: The Hsp72 level (G+/-S(G)) and olive comet tail moment (G+/-S(G) of peripheral blood lymphocytes in high exposure workers (1.24 +/- 0.42 and 4.49 +/- 1.24) were significantly higher than those in low-exposure workers (1.01 +/- 0.35 and 2.99 +/- 1.10, P < 0.05) and control (0.85 +/- 0.34 and 2.40 +/- 1.00, P < 0.05) respectively. The Hsp72 median level of all subjects was used as the limit to divide subjects into high Hsp72 level group and low Hsp72 level group. The rate with high Hsp72 level was 36.7%, 43.1% and 58.3% in control, low exposure and high exposure workers respectively and had a rising tendency following exposure level (P = 0.003). In high Hsp72 level group Hsp72 level in high exposure workers was significantly higher than that in control (P < 0.05), and there was a rising tendency along with the increase of exposed levels. But the olive comet tail moment had no significant difference among three exposed groups (P > 0.05). In low Hsp72 level group there no difference among three exposed groups about Hsp72 levels. The olive comet tail moment in high exposure workers was significantly higher than that in low exposure workers and control (P < 0.01) and high exposure workers in Hsp72 positive group and there was a rising tendency along with the increase of exposed levels. Hsp72 levels had strong negative correlation with the olive comet tail moment (r = -0.503, P < 0.01) in high exposure workers. CONCLUSION: The coke oven emissions can induce hsp72 expression. Hsp72 play a role of protecting cells from DNA damage induced by coke oven emissions. PMID- 17908428 TI - [The effect of PKC phosphorylation sites mutation in JWA coding region on TPA induced MCF-7 cell differentiation]. AB - OBJECTIVE: To investigate the effect of PKC phosphorylation sites mutation in JWA coding region on TPA-induced MCF-7 cell differentiation. METHODS: Site directed gene mutation was used to construct one or two PKC sites mutations in pEGFP-N1 JWA vectors, and transfected into MCF-7 cells by polyfect reagent, and cell differentiation was characterized by accumulation of lipid droplet as indicated by positive Oil-red-O staining of cells. RESULTS: All these transfected cell lines, MCF-7-N1(transfected with pEGFP-N1 vector), MCF-7-JWA(transfected with pEGFP-N1-JWA vector), MCF-7-JWA-1(transfected with PKC site 1 mutation pEGFP-N1 JWA vector), MCF-7-JWA-2(transfected with PKC site 2 mutation pEGFP-N1-JWA vector), MCF-7-JWA-1+2 (transfected with both PKC site 1 and 2 mutation pEGFP-N1 JWA vector) were treated with 20 nmol/L TPA for 48 h, and the percentages of positive Oil-red-O staining of cells were 48%, 67%, 69%, 67% and 70% respectively. The percentages of cell differentiation in JWA containing vectors transfected cells treated with TPA were significantly higher those of MCF-7-N1 cells (vector only control). However there were no significant differences between mutated and unmutated cells. CONCLUSION: JWA transfection enhanced MCF-7 cell differentiation induced by TPA significantly, and PKC sites mutation in JWA coding region has no obviously effect on TPA-induced MCF-7 cell differentiation. PMID- 17908429 TI - [Analysis of cell parameter in peripheral blood of occupational n-hexane exposed workers]. PMID- 17908430 TI - [Dimethylformamide induced impairment of function of liver and kidney in exposed workers and its effect on lipid metabolism]. PMID- 17908431 TI - [Application of Cox model in relationship of trinitrotoluene poisoning to length of service and type of work]. PMID- 17908432 TI - [Knowledge of occupational health and requirements for health service in workers of a steel factory]. PMID- 17908433 TI - [Coping style and psychological health in coal mine workers]. PMID- 17908434 TI - [Inhibition effect of volatile of alkyd color lacquer on superoxide dismutase in multiple organs of mice]. PMID- 17908435 TI - [Ultrastructure observation for petroleum asphalt fume induced impairment of liver and kidney in mice]. PMID- 17908436 TI - [Activity of superoxide dismutase and level of lipid peroxidation in pneumoconiosis patients]. PMID- 17908437 TI - [Recombinant human epidermal growth factor in II-degree burn caused by gas outburst]. PMID- 17908438 TI - [Distribution of types of bacteria involved in infection after burn caused by gas outburst and analysis of drug tolerance]. PMID- 17908439 TI - [Value of serum troponin I and creatine kinase isozymes in diagnosis of cardiac muscle impairment induced by acute organic phosphorus pesticide poisoning]. PMID- 17908440 TI - [Acute chlorine gas induced bronchial asthma in two patients]. PMID- 17908441 TI - [Acute organic tin poisoning in 15 patients]. PMID- 17908442 TI - [Epidemic hemorrhagic fever in laboratory personnel infected in animal experiments]. PMID- 17908443 TI - [Emergency treatment and nursing for patients with acute cyanide poisoning]. PMID- 17908444 TI - [Ion chromatography in determination of alkali metals and compounds in air of working places]. PMID- 17908445 TI - [Gas chromatography in determination of chlorpyrifos in air of working places]. PMID- 17908446 TI - [High performance liquid chromatography in determination of diethylenetriamine in air of working places]. PMID- 17908447 TI - [Double slit quartz tube flame atomic absorption spectrometry in determination of urine cadmium]. PMID- 17908448 TI - [Advance of research on cytogenetic toxicity of low-frequency electromagnetic fields]. PMID- 17908450 TI - [Analysis for problems in diagnosis and identification of occupational diseases]. PMID- 17908449 TI - [Advance of research on effect of extremely low frequency exposure in pregnant period on pregnancy]. PMID- 17908451 TI - Obstructive sleep apnea hypopnea syndrome: a proinflammatory disorder. PMID- 17908452 TI - Effect of continuous positive airway pressure treatment on serum adiponectin level and mean arterial pressure in male patients with obstructive sleep apnea syndrome. AB - BACKGROUND: Recent research suggested that obstructive sleep apnea syndrome (OSAS) might be independently associated with hypoadiponectinemia, which was linked to some complications of OSAS, such as hypertension, diabetes, etc. This study was conducted to investigate the effect of continuous positive airway pressure (CPAP) treatment on changes of both serum adiponectin levels and mean arterial pressure and their possible links in male OSAS patients. METHODS: Twenty three adult male patients with moderate-to-severe OSAS but without obesity, coronary heart disease and diabetes were recruited. Their blood samples were collected and morning mean arterial pressure (MAP) was measured before CPAP treatment and on day 3, 7, 14 of CPAP treatment respectively. The serum adiponectin concentration was tested with radioimmunoassay. RESULTS: Compared with the serum adiponectin level before CPAP treatment, no significant change was found in OSAS patients on day 3 and day 7 of CPAP treatment (P > 0.05). It was not until day 14 of CPAP treatment did a significant elevation in serum adiponectin level occur (P < 0.01). Meanwhile, the MAP showed no statistically significant difference among its levels before CPAP, on day 3 and day 7 of CPAP treatment (P > 0.05). However, on day 14 of CPAP treatment, a significantly lower MAP than that obtained before treatment was observed (P < 0.05). CONCLUSIONS: CPAP treatment can gradually reverse hypoadiponectinemia and reduce MAP in OSAS patients. Hypoadiponectinemia might be involved in the pathogenesis of OSAS mediated hypertension. PMID- 17908455 TI - Diagnosis and surgical treatment of pancreatic endocrine tumors in 36 patients: a single-center report. AB - BACKGROUND: Pancreatic endocrine tumors (PETs) are rare and their surgical treatment is often debated. The purpose of this retrospective study was to analyze the diagnosis and surgical strategy of functioning and non-functioning PETs. METHODS: From May 1980 to March 2006, 36 patients with pancreatic endocrine tumors at the Second Affiliated Hospital of Zhejiang University were retrospectively studied. RESULTS: Among the 36 patients, 29 (81%) had functioning tumors, and 7 (19%) had nonfunctioning tumors. Ninety-two percent of insulinomas were benign, whereas 4 (57%) of nonfunctioning PETs were malignant. The size of functioning tumors was (2.3 +/- 0.3) cm, that of nonfunctioning tumors was less than (5.1 +/- 0.5) cm. The combination CT and transabdominal ultrasonography resulted in a diagnostic sensitivity of 84%. Thirty-three primary lesions were precisely located in 32 patients (89%). Atypical tumor resection was performed for 73% of functioning tumors, while typical pancreatectomy was performed for 6 (85%) of nonfunctioning tumors. Moreover, 5 liver resections and 1 lymph node dissection were performed. During the follow-up, fifteen complications occurred in 12 (36%) patients after operation. The 5-year survival rate for patients with benign tumors was 92% compared to 50% for those with malignant tumors. Surgical cure was achieved in 95% of patients with benign insulinomas. CONCLUSIONS: Surgical strategy for PETs depends on the size and location of the tumor and the risk of malignancy. The optimal surgical procedure is key to prevent postoperative complication. Radical resection including initial and metastatic lesion may benefit patients with malignant PETs. PMID- 17908454 TI - Increased serum levels of C-reactive protein and matrix metalloproteinase-9 in obstructive sleep apnea syndrome. AB - BACKGROUND: Obstructive sleep apnea syndrome (OSAS), characterized by intermittent hypoxia/reoxygenation (IHR), has been identified as an independent risk factor for cardiovascular diseases (CVD). The CVD biomarkers associated with OSAS have not been thoroughly investigated. METHODS: Fifty-one men with OSAS recently diagnosed by polysomnography were classified into two groups according to the severity of apnea: moderate to severe OSAS group (n = 28) and mild OSAS group (n = 23). Twenty-five obese men, of comparable age and body mass index (BMI), without OSAS were chosen as control subjects. Serum metabolic variables, C reactive protein (CRP) and matrix metalloproteinase-9 (MMP-9) were measured. Spearman correlation and regression analysis were performed. RESULTS: Serum concentrations of CRP and MMP-9 were significantly higher in 51 OSAS patients than in 25 control subjects. Levels of CRP and MMP-9 were significantly higher in patients with moderate to severe OSAS than in patients with mild OSAS or in obese control subjects. A positive correlation was found between levels of CRP and MMP 9 in OSAS patients. Regression analysis showed that after adjusting for age and BMI, apnea/hypopnea index (AHI) significantly correlated with serum concentrations of CRP and MMP-9 in patients with OSAS. CONCLUSIONS: AHI, mirroring the frequency of IHR, was a predictor of enhanced circulating CVD biomarkers MMP-9 and CRP. Our data support the theory that IHR contributes to the upregulation of the inflammatory factors in OSAS patients. PMID- 17908457 TI - Long-term visual outcome of dense bilateral congenital cataract. AB - BACKGROUND: Dense congenital cataracts often cause severe visual impairment. The results of long-term follow-up of dense bilateral congenital cataract in China have not been well documented. The purpose of this study was to evaluate the long term visual function in children who underwent cataract extraction for dense bilateral congenital cataract in southern part of China. METHODS: Medical records of children who underwent surgery of dense bilateral congenital cataract between January 1992 and December 2000 at Zhongshan Ophthalmic Center of Sun Yat-sen University were retroactively reviewed. In 38 children available for current follow-up, best corrected visual acuity (BCVA) and stereoscopic vision, as well as nystagmus, strabismus, and other complications, were evaluated. The mean follow-up period was 107.6 months (range 60 to 167 months). RESULTS: The mean age of cataract extraction and secondary intraocular lens implantation were 5.6 months (range 3 to 12 months) and 4.2 years (range 2.4 to 15 years), respectively. The mean BCVA was 0.25 in the better eye and 0.16 in the fellow eye. Stereoscopic vision was absent in all patients, and 3 children had simultaneous perception. Nystagmus was detected in all cases and strabismus in 35 cases. A high correlation was found between timing of cataract extraction and final BCVA of the better eye (r = -0.55, P = 0.00). A statistically significant difference was found in BCVA between post- and pre-treatment of amblyopia (t = 5.65, P = 0.00). CONCLUSIONS: Long-term visual function in children with dense bilateral congenital cataract was poor when cataract surgery was performed at age of 3 months or later. Earlier cataract surgery with adequate optical rehabilitation contributed to better visual outcome. PMID- 17908456 TI - Detection of pim-1 mRNA in prostate cancer diagnosis. AB - BACKGROUND: Pim-1 plays an important role in the apoptosis, proliferation, differentiation of cancer cells and progression of cancer. In this study we detected the expression of pim-1 mRNA in normal prostate, benign prostatic hyperplasia (BPH), and prostate cancer (PCa) and explored its diagnostic value for PCa. METHODS: The prostate tissues were collected from 23 patients with PCa, 37 patients with BPH, and 3 healthy volunteers. Pim-1 mRNA expression levels in these samples were determined by the quantitative real-time PCR (QRT-PCR). The differences of expression were calculated based on a standard curve. RESULTS: The ratio of pim-1 mRNA to beta-actin in the normal prostate, BPH, and PCa were 1.05 +/- 0.04, 2.57 +/- 0.74 and 4.45 +/-0.63, respectively. The differences among PCa, BPH and NT were significant (P < 0.05, respectively). CONCLUSION: Detecting pim-1 mRNA expression by QRT-PCR provides a reliable metric for the diagnosis of PCa. PMID- 17908458 TI - Expression of peroxisome proliferator-activated receptor gamma, E-cadherin and matrix metalloproteinases-2 in gastric carcinoma and lymph node metastases. AB - BACKGROUND: Peroxisome proliferator activated receptor gamma (PPARgamma) is a ligand-activated transcription factor. Activation of PPARgamma has recently been demonstrated to inhibit various tumor cells growth, progression and metastasis. E cadherin-mediated cell adhesion system is now considered to be an "invasion suppressor system" in cancer tissues. Matrix metalloproteinases-2 (MMP-2) is a prerequisite for metastasizing tumor cells. However their correlation is still unknown in gastric carcinoma. The aim of this study was to assess the expression of PPARgamma, E-cadherin, MMP-2 and their correlation in gastric carcinoma and metastases. METHODS: Gastric carcinoma tissues and their corresponding lymph nodes with metastases and the adjacent non-tumor tissues were obtained from 54 patients with gastric cancer who underwent gastrectomy. Expression of PPARgamma, E-cadherin and MMP-2 was assessed by immunohistochemical staining. RESULTS: The nuclear expression level of PPARgamma in neoplastic cells was significantly lower than that in the normal controls (P < 0.001), with the expression of PPARgamma being weaker in primary tumors compared with that in metastases. In all neoplastic cells, E-cadherin was expressed with abnormal patterns (cytoplasm pattern, cytoplasm and membrane pattern or absent), compared with normal cells where E-cadherin was expressed with a normal pattern (membrane pattern). Compared with the normal tissues, the expression level of E-cadherin decreased in primary tumors and further decreased in metastases (P < 0.001). Membrane staining of MMP 2 was detected in the foveolar epithelia of normal gastric mucosa, whereas predominant cytoplasm staining of MMP-2 was found in malignant tissues. The expression of MMP-2 was stronger in metastatic tissues than in primary tumors. In neoplastic foci the expression of PPARgamma was negatively correlated with MMP-2 expression (P < 0.05). However, there was no correlation between E-cadherin and PPARgamma or MMP-2 expression. CONCLUSIONS: Down-regulation of PPARgamma and E cadherin and up-regulation of MMP-2 in neoplastic foci might be helpful to gastric carcinogenesis and metastases. An inverse relationship between PPARgamma and MMP-2 in human gastric carcinoma suggests that PPARgamma might modulate MMP-2 expression and affect gastric cancer metastases. PMID- 17908459 TI - Expressions of tumor necrosis factor-converting enzyme and ErbB3 in rats with chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is associated not only with airway inflammation characterized by mucin hypersecretion but also with systemic inflammation. Tumor necrosis factor alpha (TNF-alpha) is found to take part in systemic inflammation, and ErbB3 plays an important role in mucin hypersecretion of COPD. Since TNF-alpha converting enzyme (TACE) is involved in the activation of both TNF-alpha and ErbB3, we established rat models of COPD to investigate the expressions of TACE, TNF-alpha and ErbB3 and to explore the correlations among TACE, TNF-alpha and ErbB3 respectively. METHODS: Thirty Wistar male rats were randomly divided into COPD group (group C, n = 10), saline solution parallel group (group P, n = 8), and normal control group (group N, n = 8). Group C was challenged with passive cigarette smoking and intratracheal instillation of lipopolysaccharide. Six weeks later pulmonary functions were tested, bronchoalveolar fluid and arterial blood gases were assayed, and histopathological evaluations were performed in turn. The expressions of TACE, TNF-alpha and ErbB3 in lungs of all rats were determined histochemically. RESULTS: The expressions of TACE, TNF-alpha and ErbB3 were significantly higher in group C than in group N (P < 0.01). The contents of TNF-alpha in serum (P < 0.01) and bronchoalveolar lavage fluid (BALF) (P < 0.01) were elevated more significantly in group C than in group N. A positive correlation existed between TACE and TNF-alpha (r = 0.784, P < 0.01) and between TACE and ErbB3 (r = 0.526, P < 0.01) respectively. CONCLUSIONS: TNF-alpha and ErbB3 are involved in the pathogenesis of COPD. TACE contributes to the progress of COPD indirectly through the function of TNF-alpha and ErbB3. PMID- 17908460 TI - Effect of smoking cessation on airway inflammation of rats with chronic bronchitis. AB - BACKGROUND: Smoking is the major cause of airway inflammation in chronic obstructive pulmonary disease (COPD), and smoking cessation is regarded as one of the important strategies for prevention and treatment of the inflammation. The inflammation of the chronic airway may be present and deteriorated even if the COPD patients stop smoking. Whether and how early smoking cessation affects the progress of inflammation is still obscure. This study was conducted to find the appropriate time for smoking cessation to terminate the airway inflammation in rats with smoke-induced chronic bronchitis. METHODS: A rat model of COPD was established by passively inhaling smoke mixture. Fifty-four young male Sprague Dawley rats were randomly divided into 9 groups with different periods of smoke exposure and different time points of cessation. The inflammation markers to be detected included inflammatory cells in the bronchoalveolar lavage fluid (BALF), the myeloperoxidose (MPO) activity, the morphologic changes and the expression of ICAM-1 on the airway epithelium. RESULTS: When smoking was terminated at early stage, the inflammatory markers and related indexes were different from those of the typical chronic bronchitis group (group M7) (P < 0.01). The pathologic score of group SC7 (2 weeks of smoking cessation after occurrence of typical chronic bronchitis) was not different from that of group M7, and the level of ICAM-1 was still up-regulated (compared to group M7, P > 0.05). Meanwhile, most of inflammatory cells in BALF were neutrophils compared to other groups (P < 0.01). When smoking was terminated, the MPO activity was significantly lower than that of group M7 (P < 0.01). CONCLUSIONS: Smoking cessation at early stage can effectively inhibit the inflammatory reaction of COPD. Once chronic bronchitis occurs, little could be improved by smoking cessation. PMID- 17908461 TI - Active immunotherapy of allergic asthma with a recombinant human interleukin-5 protein as vaccine in a murine model. AB - BACKGROUND: Eosinophils are highly related to allergic asthma inflammation. Interleukin (IL)-5 is the major chemokine of eosinophils, inhibition of the activity of IL-5 thus seems to be a potential approach to asthma therapy. The current study was performed to determine whether a recombinant human IL-5 protein as a xenogeneic vaccine has the capability of inducing anti-asthma activities. METHODS: Recombinant human IL-5 was used as a protein vaccine. Mouse asthma model was established to observe the anti-asthma activities. Lung histology was observed; eosinophils in blood and bronchoalveolar lavage were stained and counted. Airway hyperresponsiveness was determined by whole body plethysmograph. Antibody characters and cytokines were detected with enzyme linked immunosorbent assay (ELISA) and Western blot assay. RESULTS: Vaccination with recombinant human IL-5 protein as vaccine significantly reduced airway inflammation and airway hyperresponsiveness, and shifted the cytokine production from Th2 (IL-4) to Th1 (INF-gamma) in mice allergic-asthma model. Immunization with recombinant human IL 5 protein vaccine bypassed the immunological tolerance and induced production of polyclonal antibodies that were cross-reactive with murine IL-5. CONCLUSIONS: Active immunization with xenogeneic homologous IL-5 may be a possible therapeutic approach to the treatment of asthma and potentially of other eosinophilic disorders. PMID- 17908462 TI - Effect of cigarette smoke extract on proliferation of rat pulmonary artery smooth muscle cells and the relevant roles of protein kinase C. AB - BACKGROUND: Increased proliferation of pulmonary vascular cells and muscularisation of pulmonary vessels are frequently observed in human smokers and in animals exposed to cigarette smoke. To elucidate the molecular mechanisms leading to these changes, we studied the in vitro effect of cigarette smoke extract (CSE) on proliferation of pulmonary artery smooth muscle cells (PASMCs) and activation of protein kinase C (PKC), an important kinase implicated in cell proliferation. METHODS: PASMCs cultured from 12 normal Wistar rats were studied in the following conditions: (1) PASMCs were exposed to different concentrations of CSE for 24 hours, then MTT colorimetric assay was used for detection of cell proliferation. Cell viability was assessed by trypan blue exclusion. (2) PASMCs were pre-incubated with phorbol 12-myristate 13-acetate (PMA) for 24 hours or Ro31-8220 for 30 minutes before exposure to 5% CSE for 24 hours. Cell proliferation was examined by MTT colorimetric assay, cell cycle analysis and proliferating cell nuclear antigen (PCNA) immunocytochemical staining. (3) PASMCs were exposed to 5% CSE for 24 hours. Then PKC-alpha mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) and protein expression by Western blotting, while PKC-alpha translocation was observed by immunofluorescence staining and confocal microscopy. (4) PASMCs were transfected with specific antisense oligodeoxynucleotides against PKC-alpha 6 hours before exposure to 5% CSE for 24 hours. PKC-alpha protein expression and cell proliferation were detected by methods described previously. RESULTS: (1) Low concentration of CSE (5%) increased proliferation of PASMCs, whereas high concentrations (20%, 30%) were inhibitory as a result of cytotoxicity. (2) The value of absorbance (Value A), proliferation index (PI), S-phase cell fraction (SPF) and average optical density of PCNA staining in PASMCs from 5% CSE exposure group (0.306 +/- 0.033, 0.339 +/- 0.033, 0.175 +/- 0.021, 0.315 +/- 0.038, respectively) were significantly increased compared with those of control group (0.249 +/- 0.018, 0.177 +/- 0.055, 0.092 +/- 0.023, 0.187 +/- 0.022, respectively) (P < 0.05). PKC down-regulation by PMA pretreatment or PKC inhibition by Ro31-8220 pre-incubation abolished the effect of 5% CSE on PASMCs proliferation. (3) After exposure to 5% CSE for 24 hours, PKC-alpha mRNA and protein expression in PASMCs (1.054 +/- 0.078, 1.185 +/- 0.041, respectively) were much higher than in untreated cells (0.573 +/- 0.054, 0.671 +/- 0.055, respectively) (P < 0.01). Moreover, 5% CSE induced a translocation of PKC-alpha from cytoplasm toward the perinuclear area and into the nucleus. (4) Specific antisense oligodeoxynucleotides against PKC-alpha reduced 5% CSE-induced expression of PKC-alpha protein (0.713 +/- 0.047 vs 1.180 +/- 0.056), also abolished the effect of 5% CSE on PASMCs proliferation significantly. CONCLUSIONS: CSE can be cytotoxic at high concentrations. But at low concentrations, it makes a mitogenic effect on cultured PASMCs. PKC, especially its alpha isozyme, seems to play an important role in CSE-induced proliferation of PASMC. PMID- 17908463 TI - Improving effect of Ginkgolide B on mitochondrial respiration of ischemic neuron after cerebral thrombosis in tree shrewa. AB - BACKGROUND: It has been known that platelet activating factor receptors (PAFR) may mediate many acute pathological responses and that PAFR antagonist Ginkgolide B (GB) possesses multiple effects, but the actions of GB on PAFR affinity and mitochondrial respiration in the ischemic neuron were unclear until now. This study explored the possible effects of GB on PAFR and the mitochondrial respiration of the neuron in the ischemic microenvironment. METHODS: Thrombotic cerebral ischemia in tree shrews was induced by a photochemical reaction; changes in the regional cerebral blood flow (rCBF, using (99m)Tc tracer technique), the brain water content (specific gravimetric method), PAFR (3H-labelled PAF assay), the respiratory control rate (RCR), the phosphorus-oxygen (P/O) ratio of mitochondrial respiration (Clark oxygen electrode), mitochondrial permeability transition (MPT) pore, and the mitochondrial ultrastructure in the ischemic neurons were also observed. Data were compared between the two groups (the ischemia group vs the sham group, and the ischemia group vs the GB group). RESULTS: There were high affinity and low affinity sites for PAFR on the tree threws' brain cell membranes. The varying-affinity PAFR binding sites, the respiration state III, the state IV, RCR, the P/O ratio of the mitochondria, and the rCBF all decreased markedly (respectively, P < 0.01 and P < 0.05), but the water content increased (P < 0.01) in the ischemia group after the application of cerebral thrombosis. In tree shrews treated with GB (5 mg/kg i.v.) 6 hours after photochemical reaction, their PAFR binding sites and respiratory state increased markedly. The rCBF gradually increased and the brain edema ameliorated (P < 0.01) at 24h after cerebral ischemia. There were significant differences between the ischemia group and sham group (P < 0.01). In GB treated isolated neurons' mitochondria, with or without cerebral ischemia, the energy metabolism of the mitochondria had not been changed. CONCLUSIONS: The activation of the PAFR may play an important role in the inhibition of the mitochondrial respiration and the induction of neuronal damage after cerebral thrombosis; however, GB possesses neuroprotective effects by improving mitochondrial metabolism. PMID- 17908464 TI - Role of pigment epithelium-derived factor on proliferation and migration of choroidal capillary endothelium induced by vascular endothelial growth factor in vitro. AB - BACKGROUND: Pigment epithelium-derived factor (PEDF) is expressed in several normal organs and identified as an inhibitor of neovascularization. In the present study, we investigated the effect of PEDF in an in vitro model of ocular choroidal neovascularization. METHODS: Microdissection was used to isolate the human choroidal endothelial cells (CECs), followed by the use of superparamagnetic beads (Dynabeads) coated with the CD31 antibody, which selectively binds to the endothelial cell surface. The mitogenic and motogenic effects of vascular endothelial growth factor (VEGF) on cultured choroidal capillary endothelial cells were examined in the presence or absence of PEDF (1, 10, 100, and 1000 ng/ml) using cell counts and migration assays. RESULTS: Cells bound to the beads were isolated using a magnetic particle concentrator and they were successfully cultured and characterized to be endothelial cells that possessed greater than 95% immunoreactivity to von Willebrand factor. PEDF suppressed the proliferation and migration of VEGF-induced choroidal capillary endothelial cells. However, the concentration of PEDF which we used has little effect on normal CECs. CONCLUSIONS: PEDF played an important role on the growth and migration of VEGF-stimulated choroidal endothelial cell. These findings suggest that PEDF may be an effective approach to the treatment of choroidal neovascular disorders. PMID- 17908465 TI - Current opinion on the importance of pulmonary rehabilitation in patients with chronic obstructive pulmonary disease. PMID- 17908466 TI - Distribution of natural killer cell receptors in HIV infected individuals. PMID- 17908467 TI - Effects of metoprolol treatment on a disintegrin metalloproteinase expression and extracellular matrix remodeling after myocardial infarction in rats. PMID- 17908468 TI - Plummer-Vinson syndrome associated with solid-pseudopapillary tumor of the pancreas. PMID- 17908469 TI - Pulmonary sarcoid-like granulomatosis induced by aluminum dust: report of a case and literature review. PMID- 17908470 TI - Surgical outcome and clinical follow-up in patients with symptomatic myocardial bridging. AB - BACKGROUND: Myocardial bridging with systolic compression of the left anterior descending coronary artery (LAD) may be associated with myocardial ischaemia. The clinical outcome in patients with surgical treatment for symptomatic myocardial bridging remains undetermined. This study assessed the middle- and long-term results of surgical treatment for symptomatic myocardial bridging. METHODS: From 1997 to 2006, 37,463 patients received selective coronary angiography in the Fuwai Cardiovascular Hospital, Beijing, China. Of these, 484 patients had angiographic diagnosis of myocardial bridging. Of the 484 patients, 35 underwent surgery for treatment of myocardial bridging with significant systolic arterial compression. Among the surgical treatment patients, 24 presented with other cardiac disorders, and the remaining 11 symptomatic patients with isolated myocardial bridging were included in the follow-up study. RESULTS: The angiographic prevalence of myocardial bridging was 1.3% in this study. The coronary angiographies of the 11 patients revealed myocardial bridging in the middle segment of LAD causing systolic compression > or = 75% (ranging from 75% to 90%). The mean age of patients was 48.4 years. Surgical myotomy was performed in 3 patients and coronary artery bypass grafting (CABG) in 8 patients. Eight patients were operated on with an off-pump approach and 3 with a cardiopulmonary bypass technique after median sternotomy. Conversion to on-pump CABG surgery was necessary in 1 patient because of perforation of the right ventricle. The left internal mammary artery was used in all patients with CABG. The acute clinical success rate was 100% with respect to the absence of myocardial infarction, death or other major in-hospital complications. All of the patients were followed up clinically. The median follow-up was 35.3 months (range: 6 to 120 months). Nine patients were free from symptoms and one of them continued taking beta blockers. The remaining 2 patients with myotomy had atypical chest pain. One received coronary angiography again and no stenosis was found two years after operation; while exercise testing was performed in the other patient and revealed no evidence of myocardial ischaemia. None of the patients sustained a myocardial infarction or other major adverse cardiac events (death or vessel revascularization) during follow-up. CONCLUSIONS: Myocardial bridging is a relatively common angiographic finding. Surgical myotomy or CABG should be limited to patients who are refractory to oral medication. Surgical relief of myocardial ischaemia due to systolic compression of intramyocardial coronary arteries can be accomplished with low operative risk and excellent middle- and long-term results. PMID- 17908471 TI - Combined monitoring of evoked potentials during microsurgery for lesions adjacent to the brainstem and intracranial aneurysms. AB - BACKGROUND: Neurophysiologic monitoring during surgery is to prevent permanent neurological injury resulting from surgical manipulation. To improve the accuracy and sensitivity of intraoperative neuromonitoring, combined monitoring of transcranial electrical stimulation motor evoked potentials (TES-MEPs), somatosensory evoked potentials (SSEPs) and brainstem auditory evoked potentials (BAEPs) was attempted in microsurgery for lesions adjacent to the brainstem and intracranial aneurysms. METHODS: Monitoring of combined TES-MEPs with SSEPs was attempted in 68 consecutive patients with lesions adjacent to the brainstem as well as intracranial aneurysms. Among them, 31 patients (31 operations, 28 of posterior cranial fossa tumors, 3 of posterior circulation aneurysms) were also subjected to monitoring of BAEPs. The correlation of monitoring results and clinical outcome was studied prospectively. RESULTS: Combined monitoring of evoked potentials (EPs) was done in 64 (94.1%) of the 68 patients. MEPs monitoring was impossible for 4 patients (5.9%). No complication was observed during the combined monitoring in all the patients. In 45 (66.2%) of the 68 patients, EPs were stable, and they were neurologically intact. Motor dysfunction was detected by MEPs in 8 patients, SSEPs in 5, and BAEPs in 4, respectively. CONCLUSIONS: A close relationship exists between postoperative motor function and the results of TES-MEPs monitoring. TES-MEPs are superior to SSEPs and BAEPs in detecting motor dysfunction, but combined EPs serve as a safe, effective and invasive method for intraoperative monitoring of the function of the motor nervous system. Monitoring of combined EPs during microsurgery for lesions adjacent to the brainstem and intracranial aneurysms may detect potentially hazardous maneuvers and improve the safety of subsequent procedures. PMID- 17908472 TI - T3/T4 thoracic sympathictomy and compensatory sweating in treatment of palmar hyperhidrosis. AB - BACKGROUND: Compensatory sweating (CS) is one of the most common postoperative complications after thoracic sympathectomy, sympathicotomy or endoscopic sympathetic block (ESB) for palmar hyperhidrosis. This study was conducted to examine the relevance between CS and the sympathetic segment being transected in the surgical treatment of palmar hyperhidrosis, and thus to detect the potential mechanism of the occurrence of CS. METHODS: Between October 2004 and June 2006, 163 patients with primary hyperhidrosis were randomly divided into two groups, T(3) sympathicotomy (78 patients) and T(4) sympathicotomy (85), who were operated upon under general anesthesia via single lumen intubation and intercostal video mediastinoscopy (VM). RESULTS: No morbidity or mortality occurred. Palmar hyperhidrosis was cured in all patients. Follow-up (mean (13.8 +/- 6.2) months) showed no recurrence of palmar hyperhidrosis. The difference of rates of mild CS in groups T(3) and T(4) was of no statistical significance. The rate of moderate CS was significantly lower in group T(4) than in group T(3). No severe CS occurred. CONCLUSION: The rates of occurrence and severity of CS are lowered with the lower sympathetic chain being transected. PMID- 17908474 TI - Relationship between adrenal function and prognosis in patients with severe sepsis. AB - BACKGROUND: It is known that the hypothalamic-pituitary-adrenal (HPA) axis is highlighted by stimulation, such as sepsis, trauma, etc, when corticortropin increases and plasma cortisol levels enhance. Relative adrenal insufficiency is not uncommon in critically ill patients and may occur in severe sepsis patients with high plasma cortisol levels. It has been demonstrated that a short corticotropin test has a good prognostic value and is helpful in identifying patients with septic shock at high risk for death, but it has not been established for all severe sepsis patients, especially in China. The aim of this study is to explore the relationship between adrenal function and prognosis in patients with severe sepsis. METHODS: This prospective study was conducted between July and December 2004 in 6 teaching hospitals. Two hundred and forty patients with severe sepsis were enrolled in this study. A short corticotropin stimulation test was performed in all patients by intravenous injection of 250 microg of corticotropin. Blood samples were taken immediately before the test (T0), 30 (T30) and 60 (T60) minutes afterward, and the plasma cortisol concentration was measured by radio-immunoassay. At the onset of severe sepsis, the following parameters were recorded: age, sex, Acute Physiology and Chronic Health Evaluation (APACHE) II, heart rate, mean arterial pressure (MAP), arterial partial pressure of oxygen (PaO(2))/fraction of inspired oxygen (FiO(2)), peripheral blood of hemoglobin, platelets and leukocyte concentration and the number of organ failure. Patients were designated into two groups (survival and non-survival groups) according to the 28-day mortality. Relative adrenal insufficiency was defined as the difference between T0 and the highest value of T30 or T60 (DeltaTmax) < or = 9 microg/dl. RESULTS: (1) Two hundred and forty patients with severe sepsis were included in this study, with 134 patients in the survival group and 106 in the non-survival group. The 28-day mortality was 44.2%. (2) Between the survival group and non-survival group age, APACHE II, peripheral blood of platelets, the number of organ failures, T0 and DeltaTmax showed significant differences. T0 was (23 +/- 10) microg/dl and (36 +/- 18) microg/dl in the survival group and nonsurvival group respectively. DeltaTmax was (18 +/- 9) and (10 +/- 8) microg/dl in the survival group and non-survival group respectively. The areas under the ROC curve for T0 and DeltaTmax were both 0.72, and the area under the ROC curve for APACHE II was 0.70. By multivariate analysis age, T0, the number of organ failures and relative adrenal insufficiency (DeltaTmax < or = 9 microg/dl) were independent predictors of death. (3) The incidence of relative adrenal insufficiency was 38.3% in total, 19.4% in the survival group and 62.3% in the non-survival group (P < 0.001). The 28-day mortality was 71.7% among the relative adrenal insufficiency patients but 27.0% among normal adrenal function patients. CONCLUSIONS: The prevalence of relative adrenal insufficiency is high in severe sepsis. Relative adrenal insufficiency has a good prognostic value for severe sepsis. PMID- 17908475 TI - Small hepatocellular carcinoma with peripheral enhancement: pathological correlation with dual phase images by helical CT. AB - BACKGROUND: The peripheral enhancement of small hepatocellular carcinoma (SHCC) is a rare appearance in dual phase images by helical computed tomography (CT). This study discusses this phenomenon and its correlative histopathology. METHODS: The helical CT dual phase appearance of peripheral enhancement in SHCC was analyzed in 21 cases (22 lesions). All lesions were confirmed as SHCC by histopathological examination. RESULTS: In these 22 lesions, enhanced peripheral ring in 20 lesions was incomplete, the thickness of enhanced peripheral ring varied and mural node could be found in hepatic arterial phase; only 2 lesions had complete peripheral ring enhancement and ring of uniform thickness in hepatic arterial phase. The enhancement of some peripheral rings and mural nodes dropped to very low density in portal venous phase. The tumour cells were grade I in 3 lesions, II in 16, III in 2 and IV in 1. The vascular supply was more abundant at the border than in the centre of 15 lesions and the vascular supply was deficient in both centre and border of the remaining 7 lesions. In 3 lesions, the pseudocapsule showed in the border of the lesion. In 12 lesions, flecks of necrosis were found in the border and/or centre of the lesion. CONCLUSIONS: The characteristic peripheral enhancement in helical CT dual phase images of small hepatocellular carcinoma correlates with different vascular supplies, fibrous capsule and necrosis of the lesion. PMID- 17908477 TI - Public knowledge of heart attack symptoms in Beijing residents. AB - BACKGROUND: Definitive treatment for heart attack is early reperfusion with either angioplasty or thrombolytic therapy, and the benefit is strictly time dependent. Patient outcomes are improved with either therapy when initiated as soon as possible. Recognition of heart attack symptoms is logically tied to taking action to receive prompt emergency care. Inadequate knowledge of heart attack symptoms may prolong delay. The purpose of this study was to document knowledge about heart attack symptoms in Beijing residents and to identify the characteristics associated with increased knowledge of heart attack. METHODS: A structured survey was conducted in 18 communities in Beijing from March 1 through June 10 in 2006. Addresses and participants were selected randomly following a stratification. The survey was designed to collect knowledge of heart attack symptoms from sampled adults in each community. RESULTS: A total of 4627 respondents completed the questionnaires correctly, and 50.29% of them were female. Totally 64.15% of the respondents reported chest pain or discomfort (common symptoms) as a symptom of heart attack; 75.38% reported at least one of the following eight symptoms as a symptom of heart attack: back pain, shortness of breath, arm pain or numbness, nausea or vomiting, neck, jaw or shoulder pain, epigastric pain, sweating, weakness (less common symptoms); 20.36% correctly reported four or more heart attack symptoms, only 7.4% knew all the correct heart attack symptoms, and 28.94% knew about reperfusion therapy for heart attack; 31.7% reported to call 120 or 999 while having a heart attack themselves; however 89.6% reported to call 120 or 999 when someone else is suffering from a heart attack. Very old persons and those with health insurance coverage, high education level, high household income, longer living in Beijing and previous experience with heart disease had greater knowledge of heart attack symptoms. CONCLUSIONS: Public knowledge of common heart attack symptoms as well as less common heart attack symptoms is deficient in Beijing residents. But their knowledge of calling emergency medical services when someone is having a heart attack is relatively adequate. Public health efforts are needed to increase the recognition of the major heart attack symptoms in both the general public and groups at high risk for an acute cardiac event, especially in socioeconomically disadvantaged subgroups, including persons with low education level, low household income, and no health insurance coverage. PMID- 17908478 TI - Adipocytokines and breast cancer risk. AB - BACKGROUND: Many researches suggested that obesity increased the risk of breast cancer, but the mechanism was currently unknown. Adipocytokines might mediate the relationship. Our study was aimed to investigate the relationship between serum levels of resistin, adiponectin and leptin and the onset, invasion and metastasis of breast cancer. METHODS: Blood samples were collected from 80 newly diagnosed, histologically confirmed breast cancer patients and 50 age-matched healthy controls. Serum levels of resistin, adiponectin and leptin were determined by enzyme-linked immunosorbent assays (ELISA); fasting blood glucose (FBG), lipids, body mass index (BMI), and waist circumference (WC) were assayed simultaneously. RESULTS: Serum levels of adiponectin ((8.60 +/- 2.92) mg/L vs (10.37 +/- 2.81) mg/L, P = 0.001) and HDL-c were significantly decreased in breast cancer patients in comparison to controls. Serum levels of resistin ((26.35 +/- 5.36) microg/L vs (23.32 +/- 4.75) microg/L, P = 0.000), leptin ((1.35 +/- 0.42) microg/L vs (1.06 +/- 0.39) microg/L, P = 0.003), FBG and triglyceride (TG) in breast cancer patients were increased in contrast to controls, respectively. However, we did not find the significant difference of the serum levels of resistin, adiponectin and leptin between premenopausal breast cancer patients and healthy controls (P = 0.091, 0.109 and 0.084, respectively). The serum levels of resistin, adiponectin and leptin were significantly different between patients with lymph node metastasis (LNM) and those without LNM (P = 0.001, 0.000 and 0.006, respectively). The stepwise regression analysis indicated that the tumor size had the close correlation with leptin (R(2) = 0.414, P = 0.000) and FBG (R(2) = 0.602, P = 0.000). Logistic regression analysis showed that reduced serum levels of adiponectin (OR: 0.805; 95% CI: 0.704 - 0.921; P = 0.001), HDL (OR: 0.087; 95% CI: 0.011 - 0.691, P = 0.021), elevated leptin (OR: 2.235; 95% CI: 1.898 - 4.526; P = 0.004) and resistin (OR: 1.335; 95% CI: 1.114 - 2.354; P = 0.012) increased the risk for breast cancer; Reduced serum levels of adiponectin (OR: 0.742; 95% CI: 0.504 - 0.921; P = 0.003) and elevated leptin (OR: 2.134; 95% CI: 1.725 - 3.921; P = 0.001) were associated with lymph node metastasis of breast cancer. CONCLUSIONS: The decreased serum adiponectin levels and increased serum resistin and leptin levels are risk factors of breast cancer. The low serum adiponectin levels and high serum leptin levels are independent risk factors for metastasis of cancer. The association between obesity and breast cancer risk might be explained by adipocytokines. PMID- 17908479 TI - Expression of E-cadherin, beta-catenin, cathepsin D, gelatinases and their inhibitors in invasive ductal breast carcinomas. AB - BACKGROUND: E-cadherin, beta-catenin, cathepsin D, matrix metalloproteinase (MMP) 2, MMP-9, tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 are all invasion-related proteins. The expression patterns of these proteins in invasive ductal breast carcinomas, and their associations with known clinicopathological parameters, tumor recurrence and expressions of estrogen receptor (ER), progesterone receptor (PR), PS2 and c-erbB2 were not well studied in Chinese patients. METHODS: In a set of 94 invasive ductal breast carcinomas, protein expressions of these molecular markers were investigated by immunohistochemistry, and their associations with known clinicopathological parameters, tumor recurrence and expressions of ER, PR, PS2 and c-erbB2 were also examined. In addition, the interrelationship between the expressions of these proteins were studied. RESULTS: Preserved membrane E-cadherin expression was associated with late tumor stage and tumor recurrence, whereas the reduced junctional beta catenin associated with positive lymph node status and c-erbB2 overexpression. Positive staining of cathepsin D in tumor stromal cells displayed a significant association with late tumor stage. High expression of MMP-2 in cancer cells was associated with large tumor size and PR positive expression. TIMP-2 expression was positively associated with tumor recurrence. In addition, inter-relationship between the expressions of these biomarkers was also assessed. Cathepsin D staining in cancer cells was inversely correlated with its staining in stromal cells, and also inversely correlated with MMP-2 staining in tumor stromal cells. MMP-2 expression in stromal cells displayed an inverse correlation with TIMP-2 expression. MMP-9 expression displayed parallel associations with TIMP-1 and TIMP 2 expression. CONCLUSION: Evaluation of E-cadherin, beta-catenin, cathepsin D, MMP-2 and TIMP-2 expression may be of some help in more accurately predicting the prognosis of invasive ductal breast carcinomas. PMID- 17908480 TI - Efficacy of hepatitis B immunoglobulin in relation to the gene polymorphisms of human leukocyte Fcgamma receptor III (CD16) in Chinese liver transplant patients. AB - BACKGROUND: Although the use of hepatitis B immunoglobulin (HBIG) may lead to a significant reduction in recurrent hepatitis B virus (HBV) infection and improve the survival of patients who have undergone liver transplantation (LT) for hepatitis B-related diseases, the recurrence of the disease still remains at a lower level. Different clinical curative effects were observed in patients with the same HBV-related diseases and the same therapy. This study was undertaken to investigate whether the efficacy of HBIG is associated with FCGR3A gene polymorphisms in Chinese liver transplant patients. METHODS: Altogether 77 patients who had received liver transplantation for hepatitis B-related diseases with more than one-year survival after surgery were studied. The recurrence of HBV was characterized by the appearance of HBsAg in serum after the operation. The FCGR3A genotyping was performed using genomic DNA sequencing (ABI 3037). Single nucleotide polymorphism at nucleotide 559 was detected by Polyphred. RESULTS: Of the 77 patients, 14 were complicated with HBV recurrence post transplant. The FCGR3A at nucleotide 559 TT was observed in 35 (45.5%) subjects, whereas TG in 31 (40.3%) and GG in 11 (14.3%). In the 559G carrier group (n = 42, 54.5%), the risk of HBV recurrence was 9.5%, and 1- and 2-year recurrence-free survival rates were 95.2% and 88.7%, respectively. In the 559G noncarrier group (n = 35, 45.5%), the risk of HBV recurrence was 28.6%, and 1- and 2-year recurrence-free survival rates were 74.3% and 69.3%, respectively. The risk of HBV recurrence and the recurrence-free survival rate were both statistically different between the 559G carrier and noncarrier groups (P < 0.05). CONCLUSIONS: A single nucleotide polymorphism (T/G) at position 559 of the FCGR3A gene was found in Chinese patients. The efficacy of HBIG in prophylaxis of HBV recurrence after LT is associated with the gene polymorphism, so detecting FCGR3A genotypes can be a clinical reference of the HBIG administration. PMID- 17908481 TI - Expression of oestrogen receptor-alpha and oestrogen receptor-beta in prostate cancer. AB - BACKGROUND: Recent studies have suggested that estrogens are involved in normal and abnormal prostate growth, though their exact role is still controversial. Oestrogens exert inhibitory and stimulatory effects on prostate gland, but the expression of oestrogen receptor-alpha (ERalpha) and oestrogen receptor-beta (ERbeta) in malignant prostate tissue remains unresolved. We determined ERalpha and ERbeta in prostate cancer and investigated the relationship between expression of ER and pathological features of prostate carcinoma. METHODS: Thirty two cases of prostate cancer, 12 cases of normal prostate tissue and 32 cases of benign prostate hyperplasia were analyzed for the expression of ERalpha and ERbeta using semiquantitative, reverse transcription polymerase chain reaction (RT-PCR) and the products sequenced. RESULTS: Comparisons of the normal, hyperplastic and tumour prostate tissues indicated an overexpression of ERalpha in tumour specimens (P < 0.01). However, the expression of ERbeta significantly reduced in tumour tissues compared with normal and hyperplastic specimens (P < 0.01), suggesting that severe pathological features of prostate cancer were associated with lower ERbeta expression. Spearman analysis showed negative correlation between ERbeta expression and tumour stage, grade (-0.67, -0.43, respectively, both P < 0.05), and a positive correlation between ERalpha expression and tumour stage, grade (0.51, 0.57, respectively, both P < 0.01). Our analysis also showed that hormone refractory, prostate cancer, compared with hormone dependent, prostate cancer, displayed a decreased expression of ERbeta (P < 0.01) and an increased expression of ERalpha. CONCLUSIONS: ERalpha and ERbeta may play important roles in the development of prostate cancer. The decrease in ERbeta expression is associated with higher Gleason grade tumours and prostate cancer with higher metastatic potential. The loss of ERbeta could be one of the key processes leading to uncontrolled growth of prostate epithelial cells. PMID- 17908482 TI - Intrathecal administration of resiniferatoxin produces analgesia against prostatodynia in rats. AB - BACKGROUND: Prostatodynia remains a difficult clinical problem. Resiniferatoxin (RTX), an ultrapotent vanilloid, can produce a selective and long-lasting desensitization of nociception via C-fiber sensory neurons. Substance P (SP) and calcitonin gene-related peptide (CGRP) released from C-fibers are key neurotransmitters in visceral pain. In this study, we evaluated the analgesic effect of intrathecal RTX on rat prostatodynia. METHODS: Male Sprague-Dawley rats were divided into 3 groups for different treatment. In group A, sham operation was preformed. In group B, 100 microl complete Freund's adjuvant (CFA) was injected into the rat's bilateral ventral prostate to induce chronic inflammation. In group C, after prostatitis formed, 50 microl 10 nmol/L RTX was injected into the rat's lumbosacral (L5-S2) vertebral canal. SP and CGRP contents in the spinal cord were investigated by immunohistochemistry and radioimmunoassay (RIA). Their transcriptional levels in dorsal root ganglion (DRG) were determined by reverse transcriptase polymerase chain reaction (RT-PCR). In addition, pelvic nerve afferent discharge was recorded to explore the neuro-electrophysiological mechanisms underlying RTX-induced effect. RESULTS: SP and CGRP released in the spinal cord and their synthesis in DRG were increased significantly in response to CFA-induced chronic prostatitis, whereas this increase was effectively inhibited by intrathecal RTX. Meanwhile, pelvic nerve afferent electrical activity was enhanced significantly in rats with chronic prostatitis, but it was attenuated markedly in RTX-treated rats paralleled by the change of neuropeptides. CONCLUSIONS: Intrathecal RTX administration could produce an analgesic effect on rat prostatodynia. Suppression of pelvic nerve afferent electrical activity may be a crucial mechanism underlying RTX-induced analgesia. RTX intrathecal application may present a novel analgesic strategy of prostatodynia. PMID- 17908483 TI - High efficient generation of replication-defective adenoviruses containing thymidine kinase by homogeneous recombination in bacteria. AB - BACKGROUND: Suicide gene therapy is a widely used molecular treatment for head and neck cancer. In this study, we try to use the method of homogenous recombination in bacteria to clone thymidine kinase gene (tk)-a kind of suicide gene to adenovirus backbone vectors for the construction of replication-defective adenoviruses. METHODS: pAdTrack-CMV/tk was constructed through subclone of a restriction endonuclease fragment including thymidine kinase gene from plasmid pCMV-tk to another plasmid pAdTrack-CMV, and then co-transfected with supercoiled pAdEasy-1, which was an adenoviral backbone vector except for deletions of E1 and E3, to competent E. coli BJ5183 for homogenous recombination using electroporation procedure. With the same method, pAdTrack-CMV was also co transformed with pAdEasy-1 for homogenous recombination in BJ5183. Identified with restriction endonuclease PacI and polymerase chain reaction (PCR), plasmids pAd-GFP/tk and pAd-GFP were successfully constructed. Each of them was digested with PacI and sequently transfected into human embryo kidney 293 cells (HEK293) using Lipofectamine 2000. RESULTS: Comet-like adenovirus-producing foci of Ad GFP/tk and Ad-GFP were observed after 5 to 7 days of cell culture. After twelve days of packaging, the replication-defective adenoviruses were collected. Identified with PCR, thymidine kinase gene was successfully constructed into Ad GFP/tk. CONCLUSION: The replication-defective adenoviruses containing thymidine kinase can be constructed more easily by homogenous recombination in bacteria than conventional techniques. PMID- 17908484 TI - Gene induction and apoptosis in human hepatocellular carci-noma cells SMMC-7721 exposed to 5-aza-2'-deoxycytidine. AB - BACKGROUND: Aberrant DNA methylation plays a key role in human carcinogenesis. 5 aza-2'-deoxycytidine inhibits DNA methylation and induces the expression of genes putatively silenced by promoter methylation in vitro. There are few studies of the biological and clinical significance of 5-aza-2'-deoxycytidine in human hepatocellular carcinoma. This study explored the mechanism of 5-aza-2' deoxycytidine targeting transcriptional repressor complexes affecting global gene expression in hepatocellular carcinoma cell line. METHODS: High density oligonucleotide gene expression microarrays were used to examine the effects of 5 aza-2'-deoxycytidine treatments on human hepatocellular carcinoma cell line SMMC 7721. The 5' ends of the genes upregulated or downregulated in this manner were compared with BLAST database to determine whether they might have promoter CpG islands. Flow cytometry was used to detect stages of the cell cycle and apoptosis of SMMC-7721 after being treated with 5-aza-2'-deoxycytidine. RESULTS: Data obtained 3 days after 4 days of treatment with 5-aza-2'-deoxycytidine showed that more genes were induced in tumorigenic cells including genes that function in cell proliferation, differentiation, regulation of transcription, and cytokine signalling. Approximately 30% of induced genes did not have CpG islands within their 5' regions, suggesting that some genes activated by 5-aza-2'-deoxycytidine may not result from the direct inhibition of promoter methylation. This phenomenon may contribute to a number of upregulated genes involving regulation of transcription in the treated cell. Results showed that 100 micromol/L 5-aza-2' deoxycytidine blocked cell cycle at S/G2-M phase increasing rate of apoptosis. Notably, we found differential expression of molecular action in the methylation although DNA methyltransferases did not show significant difference in the treated cell line. CONCLUSION: 5-aza-2'-deoxycytidine could restore some silenced genes expression independently of DNA methylation inhibition and expression of DNA methyltransferases. PMID- 17908485 TI - Correlation among obstructive sleep apnea syndrome, coronary atherosclerosis and coronary heart disease. PMID- 17908486 TI - Detection of two amino acid deletions in HIV-1 Nef protein from Chinese former paid blood donors. PMID- 17908487 TI - Expression of protein kinase C isoforms in retinoic acid-induced differentiation of mouse embryonic stem cells into neuron-like cells. PMID- 17908488 TI - Long-term outcomes of combined chemotherapy in chronic refractory idiopathic thrombocytopenic purpura. PMID- 17908489 TI - Genetic heterogeneity of noncompaction. PMID- 17908491 TI - [Comparison of the binding affinity of LAIR-1(CD305) and LAIR-2(CD306) with their ligands]. AB - AIM: To compare the binding affinity of LAIR-1 and LAIR-2 with their ligands. METHODS: The ligand-expressing cells were incubated with the LAIR-1 and/or LAIR-2 fusion proteins at different concentration at the same time or in succession. Then the interaction changes and competition were measured by influorescent staining and flow cytometric analysis with the specific mAbs against LAIR-1 or LAIR-2. RESULTS: The membrane ligand for LAIR-1 and LAIR-2 was expressed extensively and ligand recognition was of cross-species. LAIR-2 blocked the interaction of LAIR-1 with its ligand but LAIR-1 didn't block the interaction of LAIR-2 with its ligand. CONCLUSION: LAIR-1 and LAIR-2 probably bind to the same ligand with different affinity, which provides some significant evidence for investigating the molecular mechanisms of LAIR-1 family in modulating immune response. PMID- 17908492 TI - [Secretion expression in Pichia pastoris and immunogenicity comparison of two forms of HBsAg/GM-CSF fusion proteins]. AB - AIM: To express two fusion forms of hepatitis B virus surface antigen (HBsAg-s and s-HBsAg) in the Pichia pastoris expression system, and compare immunogenicity of the two fusion proteins. METHODS: Was fused GM-CSF to 5' or 3' terminal of HBsAg by inserting the gene fragment of connecting peptide (Gly(4)Ser)(3) to linker gene of GM-CSF and HBsAg. The two fusion proteins were expressed by secreting type expression plasmid pPIC9K in the Pichia pastoris, then the expressed products were detected by SDS-PAGE, Western blot and purified by DEAE Sepharose Fast Flow ion exchange columns. Mice were inoculated with the two purified HBsAg/GM-CSF fusion proteins and HBsAg respectively in each, and the levels of anti-HBsAg in mice sera were tested by ELISA. RESULTS: Two HBsAg/GM-CSF fusion proteins were successfully expressed in the form of secretion in Pichia pastoris strain GS115, and exhibited specific reaction with both anti-HBsAg and anti-GM-CSF antibodies in Western blot. ELISA results showed after the inoculation the levels of anti-HBsAg induced by the two HBsAg/GM-CSF fusion proteins was higher than by HBsAg alone (P<0.05). Furthermore, the effect by fusing GM-CSF to C terminal of HBsAg was better than by fusing GM-CSF to N terminal of HBsAg. CONCLUSION: The immunogenicity of HBsAg could be enhanced by fusing GM-CSF. PMID- 17908493 TI - [Construction of plasmid stably expressing shRNA and inhibition of HBV replication and expression]. AB - AIM: To construct the vector stably expressing small hair RNA(shRNA) and investigate the effect of shRNA expressed with DNA vector on HBV replication and expression. METHODS: The vector expressing shRNA designated pU6 was constructed by U6 promoter. The sequence of shRNA targeting on specific region of HBV genome against HBV was designed and synthesized, then cloned into pU6. The plasmid designated pU6B/HBVi was transfected into 2.2.15 cells using Lipofectamine 2000 reagent. The HBsAg and HBeAg in the supernatants of the transfected cells were measured by ELISA. The HBV copies in the transfected cells were measured by quantitative PCR. RESULTS: Compared with controls, not only concentration of HBsAg and HBeAg but also HBV copies in transfected 2.2.15 cells decreased significantly by the plasmid expressing shRNA against HBV. CONCLUSION: HBV replication and expression in 2.2.15 cells are inhibited by stably expressed shRNA. PMID- 17908494 TI - [Inhibitory effect of short hairpin RNA on expression of Hax-1 gene in HeLa cells]. AB - AIM: To construct the short hairpin RNA (shRNA) eukaryotic expression vector specific for HS1-associated protein X-1 (Hax-1) and investigate the inhibitory effect of it on Hax-1. METHODS: According to the design rules of shRNA, the specific sequence of 19 nucleotides were selected from Hax-1 cDNA sequence and designed as the cDNA template of siRNA. ShRNA vector pGenesil-Hax-1 was constructed by recombining the synthesized specific sequence with siRNA expression vector pGensil-1. After the recombinant plasmids were transfected into HeLa cells, RT-PCR technique and Western blot were applied to analyze mRNA and protein expression of Hax-1. RESULTS: The results of RT-PCR showed that the down regulation of Hax-1 mRNA expression was found in the pGenesil-Hax-1 transfected group, but not in the pGenesil-1 transfected group or the negative control group (P<0.01). The expression of Hax-1 protein decreased by 70% in the pGenesil-Hax-1 transfected group compared with the negative control group. CONCLUSION: Hax-1 gene expression can be inhibited markedly by specific shRNA in HeLa cells, which establishes the experimental foundation for further study on the biological functions of Hax-1 in HeLa cells. PMID- 17908495 TI - [Construction, expression and purification of UreB-Omp11 fusion protein of Helicobacter pylori and its immunocompetence]. AB - AIM: To construct H.pylori vaccine candidate strain expressing UreB-Omp11 recombinant fusion protein of H.pylori. To express and purify the fusion protein UreB-Omp11 and to determine its immunocompetence. METHODS: The two genes were amplified by PCR, and the fusion gene ureB-omp11 was amplified by over lap extension PCR and then cloned into the fusion expression vector pET30a(+), pET28a(+) and pMAL-c2X. The appropriate expression system was selected, and the recombinant UreB-Omp11 fusion protein was expressed and indentfied by SDS-PAGE and Western blot analysis. Then the fusion protein was purified by MBP affinity chromatography and the purity was indentfied by SDS-PAGE. Then the fusion protein was immunized to mice. The immunized mice sera were analyzed by Western blot with purified fusion protein. RESULTS: The ureB-omp11 fusion gene was correctly insected into pET30a(+) and confirmed by Enzyme digestion and sequencing analysis; Results in SDS-PAGE and optical density scanning demonstrated that this fusion protein MBP-UreB-Omp11 was expressed in the recombinant strain of E.coli TB1(pMAL-ureB-omp11). The fusion protein UreB-Omp11 was recognized by the mice sera immunized by H.pylori, the human sera infected with H.pylori and The purity of fusion protein was 90% after purification. The fusion protein purified could be recognized by corresponding antibody of mice sera immunized by this fusion piotein, This fusion protein has strong immunoantigenicity and immunoreactivity. CONCLUSION: The prokaryotic expression system TB1 (pMAL-c2X-ureB-omp11) was successfully constructed and selected. The results obtained lay the foundation for research on development of protein and DNA vaccine of Hp. PMID- 17908496 TI - [Changes of cytokines of splenocytes in mice immunized by mix recombinant BCG EmII/3 and BCG-Em14-3-3 vaccine of Em]. AB - AIM: To investigate the changes of cytokines of splenocytes in mice immunized by mix recombinant BCG-EmII/3 and BCG-Em14-3-3 vaccine of Echinococcus multilocularis(Em) and challenged by Em protoscoleces. METHODS: BALB/c mice were vaccinated subcutaneously and intranasally by the vaccine respectively. In the eighth week of vaccination they were Challenged by Em protoscoleces.In the eighteenth week of infection they were killed for spleen. After splenocytes were separated,their culture was stimulated by EmAg or ConA and their supernatants were gathered. The levels of IL-2, IFN-gamma, TNF-alpha and IL-4 were measured by ELISA kit. Blank vector, BCG and PBS were served as control. RESULTS: In the groups of vaccine immunization, the levels of IFN-gamma and TNF-alpha increased obviously but the level of IL-4 decreased. The level of TNF-alpha in subcutaneous group was higher than that in intranasal group. CONCLUSION: Th1 response was induced in mice immunized by mix recombinant BCG-EmII/3 and BCG-Em14-3-3 vaccine of Em against the challenge by Em protoscoleces. Subcutaneous injection with this vaccine may be a good way of vaccination. PMID- 17908497 TI - [The role of cytokines in the production of IL-17 and IFN-gamma via the induction of normal human peripheral blood mononuclear cells and CD4(+) T cells]. AB - AIM: To investigate the role of cytokines (IL-23, IL-2 and IL-15) in the production and regulation of IL-17 and IFN-gamma via the induction of normal human peripheral blood mononuclear cells (PBMCs) and CD4(+) T cells. METHODS: PBMCs or CD4(+) T cells from normal human blood were cultured with IL-23, IL-2 or IL-15 in the presence or absence of IL-23. The level of IL-17 and IFN-gamma in the culture supernatants was assessed by ELISA. The frequency of IL-17 and IFN gamma produced cells was detected by ELISPOT. RESULTS: IL-23 induced PBMCs to produce IL-17 and IFN-gamma. IL-2 and IL-15 induced the production of IL-17 and IFN-gamma in a dose dependent manner by PBMCs. Similar results were confirmed by ELISPOT assay. The addition of Th2 cytokines (IL-4 and IL-10) or anti-IL-12Rbeta1 mAbs resulted in the inhibition of IL-17 and IFN-gamma production induced by IL 23. Further study suggested that IL-23, IL-2 and IL-15 directly induced the production of IL-17 and IFN-gamma by purified CD4(+) T cells. CONCLUSION: IL-23, IL-2 and IL-15 can directly induce CD4(+) T cells to produce IL-17 and IFN-gamma. The production of IL-17 and IFN-gamma induced by IL-23 can be inhibited by Th2 cytokines and anti-IL-12Rbeta1 mAbs. The finding of our research may provide some new targets for the study of mechanism and treatment of IL-17-mediated autoimmune diseases. PMID- 17908498 TI - [Immature dendritic cells directional differentiated from murine hemopoietic stem cells and morphous and function of them by MACS]. AB - AIM: To Investigate a method that can obtain massive highly purified immature dendritic cells(imDCs) steadily in vitro, and identify them by morphous, function and surface markers by MACS. METHODS: Isolate and purify CD117(+) hemopoietic stem cells(HSCs) from bone marrow of healthy C57 murine by MACS. After being expanded by SCF+IL-3, HSCs would be directional differentiated into imDCs by use of cytokine scheme of GM-CSF+IL-4+IL-10. Then identify imDCs through following ways: observing morphous and function of them under inverted microscope, scanning electron microscope and transmission electron microscope, detecting the expression of surface markers by flow cytometry. RESULTS: The fold of expansion 3, 5 and 7 days after cultured with SCF+IL-3 was separately 10.34 +/- 1.43, 22.65 +/- 2.71 and 54.39 +/- 3.08. HSCs can be successfully differentiated into imDCs, which have the function of phagocytosis. The imDCs were short and small, in shape of sentus. The expression of surfacee markers was CD11c(+), I-A/I-E(low), CD40( ), CD80(-), CD86(-) by flow cytometry. CONCLUSION: This method can obtain and identify massive highly purified imDCs steadily in vitro. PMID- 17908500 TI - [Culture and purification of olfactory ensheathing cells from human fetal using different attachment rates combined with the technique of using NT3 intermittently]. AB - AIM: To explore a simple and pragmatic method to obtain sufficient olfactory ensheathing cells from human fetal by using different attachment rates in harvested cells with the combinating of the technique of using NT3 intermittently. METHODS: DMEM/F12 culture solution including 100 mL/L of fetal bovine serum or including NT3 was used to culture olfactory ensheathing cells intermittently every 48 h. The conditions and growth degree of OECs were observed, and P75 immunocytochemistry was used to estimate the purity of the cells. RESULTS: Human fetal OECs were positive after P75 immunocytochemistry. They appeared to be dipolar or tripolar cells and their processes formed a network in vitro. The purity of OECs in good conditions reached about 95% on 9 d and 83% on 12 d. CONCLUSION: The method of using different attachment rates combined with the technique of using NT3 intermittently can culture and purify OECs simply and effectively. PMID- 17908499 TI - [Immune response induced by Mycobacterium tuberculosis Rv2450 protein in mice]. AB - AIM: To evaluate humoral and cellular immune response induced by Rv2450 protein. METHODS: Rv2450 protein was transferred to membrane and used to immunize C57BL/6 mice three times at 2 week interval. The spleen lymphocytes of the immunized mice were separated and the stimulation index (SI) was measured by MTT colorimetry and the levels of secreted IFN-gamma, IL-10 and IL-12 upon antigen-specific stimulation was detected by ELISA. RESULTS: The titer of specific antibody in sera of the immunized C57BL/6 mice was 1:3 200. The SI of Rv2450 protein immunized group (3.76 +/- 0.19) was significantly higher than that of saline immunized group (0.89+/-0.17). The IFN-gamma, IL-10, IL-12 levels in culture supernatant of spleen lymphocytes from the immunized mice was (1 740 +/- 19) ng/L, (678 +/- 15) ng/L and (469 +/- 13) ng/L respectively, significantly different from that of saline-immunized group. CONCLUSION: Rv2450 protein can be used as a candidate for the new TB vaccine. PMID- 17908501 TI - [Study on the role of Wnt/beta-catenin signaling transduction pathway in hepatocellular carcinoma cell line HepG2 and L02 cell line]. AB - AIM: To explore the possible mechanism of action of Wnt/beta-catenin signaling pathway in hepatocarcinogenesis by investigating the different expressions of the main members on this signaling pathway in hepatocellular carcinoma cell line HepG2 and L02 cell line. METHODS: The mRNAs of Wnt1, Wnt4, beta-catenin, cyclinD1 and c-myc genes were amplified by means of semiquantitative reverse transcription polymerase chain reaction (RT-PCR) in normal liver cell line L02 and hepatocellular carcinoma cell line HepG2, respectively. At the same time, the proteins expression of beta-catenin which was the key member in the Wnt/beta catenin signaling pathway was examined by immunocytochemical method and Western blot technique. RESULTS: In normal liver cell line L02, the mRNAs of Wnt1, Wnt4, cyclin D1 and c-myc genes were not detected except for the gene of beta-catenin. In hepatocellular carcinoma cell line HepG2, the mRNAs of Wnt1, beta-catenin, cyclin D1 and c-myc genes were detected except for the gene of Wnt4. Meanwhile, found that beta-catenin proteins were accumulated in the cytoplasm and/or nucleus in HepG2 but only in cell membrane in L02.Using Western blot technique, found that beta-catenin proteins expression was higher in HepG2 than in L02. CONCLUSION: The Wnt/beta-catenin signaling transduction pathway is activated with aberrant expression of Wnt1 in hepatocellular carcinoma cell line HepG2. PMID- 17908502 TI - [Immunologic tolerance of anti-Abeta antibody in the sera of AD patients]. AB - AIM: To characterize anti-amyloid-beta antibodies in the sera of Alzheimer's disease(AD) patients. METHODS: A special tissue amyloid plaque immunoreactivity (TAPIR) examinations of brains of Tg2576 mice was carried out. Abeta(42)-GST fusion protein was detected by Western blot. and then the survival rate of PC12 cells incubated with the sera and Abeta(42) was measured by MTT assay. RESULTS: The sera in AD patients could not recognize the senile plaque in the brains of Tg2576 mice and could not cross-react with Abeta(42)-GST fusion protein. Furthermore, PC12 cells incubated with the sera from AD patients presented the decreased A value compared with the healthy elderly(P<0.01). CONCLUSION: The anti Abeta antibodies in the sera of AD patients may have immunologic tolerance to amyloid peptide. PMID- 17908503 TI - [Effect of hemoperfusion treated plasma from patients with chronic severe hepatitis on the activity and expression of CYP4503A in C3A cells]. AB - AIM: To investigate the changes of CYP4503A, a key enzyme of diazepam metabolism, and explore the effect of hemoperfusion treated plasma from patients with chronic severe hepatitis on the activity and expression of CYP4503A in C3A cells. METHODS: Plasma was prepared and C3A cells were cultured. There were four groups in the experiment: normal fetal bovine plasma (NFBP) group, normal human plasma (NHP) group, hemoperfusion plasma (HPP) group, chronic severe hepatitis plasma (CSHP) group. The activity of erythromycin N-demethylase (ERD), namely the activity of CYP4503A, was measured by spectrophotometer and the expression of CYP4503A4 was detected by Western blot. RESULTS: The activity of ERD in CSHP group and HPP group was lower than that in NFBP group (P<0.05) and NHP group (P<0.05) but the activity of ERD in HPP group was higher than that in CSHP group (P<0.05). The expression of CYP4503A4 increased in NFBP group (P<0.05) and NHP group (P<0.05) but the expression of CYP4503A4 in CSHP group decreased compared with HPP group(P<0.05). CONCLUSION: The activity of ERD and the expression of CYP4503A decreased in CSHP group while the activity of ERD and the expression of CYP4503A in HPP group increased, which may cause the changes of the diazepam metabolic rates of C3A cells. This finding will provide some references for future researches on cell construction. PMID- 17908504 TI - [A dual effect on the growth of astrocytes of lipopolysaccharide]. AB - AIM: To investigate the changes of growth of astrocytes administrated with lipopolysaccharide (LPS) and the probable mechanisms. METHODS: LPS was administrated to rat astrocytes for 1 h, 6 h, 24 h and 48 h, and the changes of growth of astrocytes were detected by MTT method. Meanwhile, the long-term changes of growth of astrocytes after administration with LPS for 24 h were also studied. Moreover, the effects of inhibitor of NF-kappaB(SN50) on proliferation of astrocytes were observed. RESULTS: Changes of astrocytes growth were observed only when LPS had been administrated for 24 h. LPS of low concentration could promote proliferation of astrocytes and increase the cell viability(P<0.05 vs control), while LPS of high concentration inhibited the proliferation of astrocytes. Being administrated for 24 h, LPS could promote the proliferation of astrocytes in short-term. As for the long-term effects, LPS inhibited the proliferation of astrocytes in a concentration-dependent manner. Pretreatment with SN50 could block the effects of LPS on astrocytes. CONCLUSION: LPS of low concentration could promote the proliferation of astrocytes in short-term and LPS of high concentration could inhibit it. The molecular mechanism of the changes might be related to the activation of NF-kappaB. PMID- 17908505 TI - [Construction and primary application of two HBV-HLA-A*2402-peptide tetramers]. AB - AIM: To construct two soluble HLA-A*2402-peptide tetramers and detect the HBV specific cytotoxic T lymphocytes (CTLs) with the constructed HLA-A*2402-peptide tetramers. METHODS: After proteins HLA-A*2402-BSP and beta2m were made an effective prokanyotical expression, the purified proteins were refolded into HLA A*2402-beta2m-peptide complexes in the presence of two antigenic peptides (Hepatitis B virus Pol756-764 or Core117-125) with dilution method. Then the complexes were biotinylated by BirA enzyme and purified by gel-filtration chromatography. The tetramers were generated by mixing the complex with PE Streptavidin in the molar ratio of 5:1. FCM can and cell quest software were used to analyze the stained PBMCs. RESULTS: Two biotinylated HBV-HLA-A*2402-peptide complexes were identified by Western blot and they were shown to have natural conformation by Dot-ELISA and ELISA. CONCLUSION: The constructed HLA-A*2402 peptide tetramers can detect the HBV-specific CTLs in the patients with self limited acute HBV infection. PMID- 17908506 TI - [Construction and expression of diabody [CD3 x Pgp] without Etag]. AB - AIM: To construct and express a diabody [CD3 x Pgp] without Etag and analyse its biological activity. METHODS: In this study, the diabody [CD3 x Pgp] was obtained by PCR and restriction cleavage, and expressed in E.coli 16C9. The product was purified by anti-anti-CD3 scFv affinity chromatography and verified through SDS PAGE electrophoresis. Flow cytometry(FCM) was used to analyse the bingding properties and competitive bingding capacity. RESULTS: The sequence of diabody [CD3 x Pgp] without Etag was correct. It migrated as two bands with the expected molecular weight(25 kD and 26 kD) in SDS-PAGE. The binding rate to CD3 and Pgp antigen was 83.95% and 89.87% respectively. The competitive bingding rate to CD3 and Pgp was 43.78% and 50.25% respectively. CONCLUSION: The diabody [CD3 x Pgp] without Etag has been successfully constructed, expressed and purified. The product can bind to CD3 and Pgp antigen specifically, and its biological activity doesn't decrease. PMID- 17908507 TI - [Recombinant expression of human fibrogenic factors and preparation of polyclonal antibodies]. AB - AIM: To express the recombinant proteins of human fibrogenic factors and prepare the polyclonal antibodies of rabbit-anti-human fibrogenic factors. METHODS: The human fibrogenic factors were variant splicing forms of one gene. Their consensus sequence (ChS) was cloned into plasmid pQE30 to construct recombinant prokaryotic expression system. The recombinant expression vectors were transformed into E.coli M15, and then the expression vectors were induced by IPTG and many insoluble inclusion body proteins was obtained. The obtained proteins were used to immunize rabbits to obtain polyclonal antiserum with high titer and specificity. The eukaryotic expression of the recombinants of human fibrogenic factors was detected by Western blot. RESULTS: The insoluble inclusion body proteins were also used to prepare polyclonal antibodies with high titer and specificity. CONCLUSION: The expression of the recombinant proteins of human fibrogenic factors and the preparation of polyclonal antiserum provide a useful tool for further study of the expression and function of human fibrogenic factors. PMID- 17908508 TI - [Development and characterization of monoclonal antibodies against H7 hemagglutinin of avian influenza virus]. AB - AIM: To prepare monoclonal antibodies (mAbs) against the hemagglutinin protein of H7 subtype of avian influenza virus (AIV). METHODS: (6-8 weeks old) BALB/c mice of were immunized endermicly with H7 subtype of AIV. The splenocytes from the immunized mice were fused with Sp2/0-Ag-14 myeloma cells after the last immunization. Hybridoma cells were screened by hemagglutination (HA) and hemagglutination inhibition (HI) tests. The reactivity and specificity of mAbs were evaluated by HI test and Western blot assay. RESULTS: Four hybridoma cell lines secreting specific mAbs named 2E2, 2A4, 5F5 and 7G5 were developed. The HI titers of these mAbs were 5 x 2(7)-5 x 2(11), and the immunoglobulin subclass of 2E2 was IgM, that of 2A4 was IgG1, and that of 5F5 and 7G5 was IgG2a. Western blot analysis confirmed that the mAbs only reacted with M(r) 75 000 HA protein of H7 subtype of AIV but did not react with the proteins of Newcastle disease virus (NDV). The results of HI reactivity assay suggested that 2E2, 5F5 and 7G5 only reacted with H7 subtype of AIV but did not react with other subtypes of AIV, NDV and infectious bronchitis virus (IBV). However 2A4 reacted not only with H7 subtype of AIV but also with H15N8 reference strain of AIV at low HI level. CONCLUSION: These mAbs can be used as a useful tool to analyze the HA structure of AIV. They also provide the effective reagents for the rapid detection of H7 subtype of AIV. PMID- 17908510 TI - Newborn care: 12 beliefs that shape practice (but should they?). PMID- 17908511 TI - Acute onset of rash and oligoarthritis. PMID- 17908512 TI - Smoking cessation: tactics that make a big difference. PMID- 17908513 TI - Flu vaccination rates: how can you do better? PMID- 17908515 TI - Clinical inquiries. Which nondrug alternatives can help with insomnia? PMID- 17908514 TI - Genetic screening for iron overload: No evidence of discrimination at 1 year. AB - PURPOSE: This study measured the extent of insurance and employment problems associated with population screening for hereditary hemochromatosis and iron overload. METHODS: 101,168 primary care patients from the US and Canada were screened for iron phenotypes and HFE genotypes associated with hemochromatosis. Those identified to be at risk (2253) were offered a clinical examination, which 1677 (74%) accepted, and the 1154 of these who responded to an initial questionnaire about psychosocial issues were surveyed 1 year later about whether they had experienced problems with insurance or employment that they attributed to hereditary hemochromatosis and iron overload. RESULTS: 832 (72.1%) of the 1154 participants surveyed after 1 year responded to the second survey. Three (0.4%) had verified problems with insurance or employment that they believed were related to hereditary hemochromatosis and iron overload. Two had problems with life insurance, and one with long-term care insurance. All 3 had elevated iron levels but not a relevant HFE genotype. One of the life insurance problems was resolved; the second one was not serious. The participant who was denied long term care insurance had other health conditions unrelated to hereditary hemochromatosis and iron overload that could have contributed to the denial. No problems were verified for health insurance or employment, or from any of the comparison group participants (controls and those with inconclusive screening results). CONCLUSIONS: The risk of insurance or employment problems 1 year after phenotype and genotype screening for hereditary hemochromatosis and iron overload is very low. PMID- 17908516 TI - Clinical inquiries. Which tool is most useful in diagnosing bipolar disorder in children? PMID- 17908517 TI - Clinical inquiries. How do exercise and diet compare for weight loss? PMID- 17908518 TI - Clinical inquiries. What is the best approach to a solitary pulmonary nodule identified by chest x-ray? PMID- 17908519 TI - Clinical inquiries. How should you further evaluate an adult with a testicular mass? PMID- 17908520 TI - Sudden coma due to acute bilateral M1 occlusion. PMID- 17908521 TI - Staphylococcus aureus bacteremia and infective endocarditis: old questions, new answers? PMID- 17908522 TI - Risk factors for infective endocarditis and outcome of patients with Staphylococcus aureus bacteremia. AB - OBJECTIVE: To investigate the risk factors for Staphylococcus aureus infective endocarditis (SAIE) and 6-month mortality in patients with S aureus bacteremia (SAB). PATIENTS AND METHODS: This study consisted of patients who were diagnosed as having nosocomial or community-acquired SAB or SAIE between June 1, 2000, and December 31, 2005. Clinical characteristics of patients with SAB were compared with those of patients with SAIE, and predictors of mortality in patients with SAB were analyzed. RESULTS: The median age of the 132 randomly selected patients with SAB and the 66 patients with SAIE was 66 and 68 years, respectively. Univariable analysis showed that unknown origin of SAB, a valvular prosthesis, a pacemaker, persistent fever, and persistent bacteremia were significantly associated with SAIE. In multivariable analysis, unknown origin of SAB (odds ratio [OR], 4.2; 95% confidence interval [CI], 1.9-9.3; P=.001), a valvular prosthesis (OR, 9.2; 95% CI, 3.2-26.2; P<.001), persistent fever (OR, 3.1; 95% CI, 1.0-9.0; P=.04), and persistent bacteremia (OR, 6.8; 95% CI, 2.3-20.2- P=.001) were independently associated with SAIE. Six- month mortality was 8% in patients with SAB vs 35% in patients with SAIE (OR, 6.5; 95% CI, 2.9- 14.8; P<.001). In univariable analysis, methicillin- resistant S aureus (OR, 7.2; 95% CI, 1.7 - 29.4; P=.005) was significantly associated with 6-month mortality in patients with SAB. CONCLUSION: Unknown origin of SAB, a valvular prosthesis, persistent fever, and persistent bacteremia were independently associated with SAIE in patients with SAB. In univariable analysis, methicillin-resistant S aureus was associated with 6-month mortality in patients with SAB. S aureus infective endocarditis had a significantly higher mortality than SAB. The optimal management of SAB and SAIE deserves further study. PMID- 17908523 TI - Open-label study of a proprietary treatment program targeting type A gamma aminobutyric acid receptor dysregulation in methamphetamine dependence. AB - OBJECTIVE: To evaluate the safety and efficacy of the pharmacological component of a proprietary medical treatment program targeting type A gamma-aminobutyric acid receptor dysregulation in adults who met the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision criteria for methamphetamine dependence. PARTICIPANTS AND METHODS: A prospective, open-label, single-group study of the medication portion of a proprietary treatment program for methamphetamine dependence was conducted from July 1, 2005, to May 10, 2006, at Research Across America, an outpatient private, for-profit, clinical research company in Dallas, TX. In the study, flumazenil, hydroxyzine, and gabapentin, all of which were approved by the US Food and Drug Administration for indications other than drug dependence, were used off-label for the treatment of methamphetamine dependence. Fifty persons who had used methamphetamine within 7 days of study entry were enrolled and received the treatment. Treatment lasted 4 weeks, followed by 8 weeks of weekly follow-up visits to monitor for methamphetamine use via urine drug tests and self-reporting. RESULTS: Participant retention was higher than expected, with 85% of participants completing the program. Significant decrease in methamphetamine use (P<.001) was noted at 84 days after vs 90 days before treatment. If missing data are counted as days of methamphetamine use, a 47% reduction in use was observed for the entire sample (P<.001) and a 65% reduction for the 36 who completed the 8-week evaluation phase (P<.001). Urine test results and self-reported use were positively correlated (Pearson r=0.72, P<.001). The frequency of cravings was reduced on average by 66% (P<.001), with 30 of 31 (97%) of the 36 who completed the study reporting reduction in cravings. CONCLUSION: Substantial reductions in methamphetamine cravings and use were observed in all phases of treatment, and the retention rate of participants was high. These findings suggest that the efficacy of the medications and of the entire program in treating methamphetamine dependence should be examined in controlled trials. PMID- 17908524 TI - Long-term results of response to therapy, time to progression, and survival with lenalidomide plus dexamethasone in newly diagnosed myeloma. AB - OBJECTIVE: To determine the long-term effects of a combined regimen of lenalidomide and dexamethasone (Rev-Dex) on time to progression, progression-free survival, and overall survival (OS) in patients with multiple myeloma. PATIENTS AND METHODS: From March 2004 through October 2004, 34 patients were registered for the study. They were treated with 25 mg/d of lenalidomide on days 1 through 21 of a 28-day cycle and 40 mg/d of dexamethasone on days 1 through 4, 9 through 12, and 17 through 20 of each cycle. After 4 cycles of therapy, patients were allowed to discontinue treatment to pursue autologous stem cell transplant (SCT). Treatment beyond 4 cycles was permitted at the physician s discretion. RESULTS: Thirteen patients proceeded to SCT after initial therapy and were censored at that time point for purposes of calculation of response. Thirty-one patients achieved an objective response, defined as a partial response or better (91%; 95% confidence interval, 79%-98%), with a complete response plus very good partial response rate of 56%. The complete response plus very good partial response among the 21 patients who received Rev-Dex without SCT was 67%. The 2-year progression free survival rates for patients proceeding to SCT and patients remaining on Rev Dex were 83% and 59%, respectively; the OS rates were 92% and 90% at 2 years and 92% and 85% at 3 years, respectively. The 3-year OS rate for the whole cohort was 88%. CONCLUSION: The Rev-Dex regimen is highly active in the treatment of newly diagnosed multiple myeloma. Responses are durable with a low progression rate at 2 years. Randomized trials that incorporate quality-of-life measures are needed to determine if this and other combination regimens are better used early in therapy or should be reserved for later interventions. PMID- 17908526 TI - Viruses as an etiology of obesity. AB - Obesity is a serious chronic disease that has numerous etiologies. The prevalence of obesity has increased dramatically since about 1980 in the United States and worldwide in both developed and developing countries. This rapid spread is compatible with an infectious origin. This review discusses the 5 animal viruses and 3 human viruses that have been shown to cause obesity and examines the evidence to date for virus-induced obesity. The obesogenic animal viruses include canine distemper virus, Rous-associated virus type 7, Borna disease virus, scrapie agent, and SMAM-1. The first 4 viruses attack the central nervous system to produce obesity. SMAM-1, an avian adenovirus from India, acts directly on adipocytes and is the only animal virus that is associated with human obesity. The 3 human adenoviruses, adenovirus (Ad) 36, Ad-37, and Ad-5, that are associated with obesity also affect adipocytes directly. These viruses stimulate enzymes and transcription factors that cause accumulation of triglycerides and differentiation of preadipocytes into mature adipocytes. Ad-5 and Ad-37 have been shown to cause obesity in animals. Ad-36 has been studied the most and is the only human adenovirus to date that has been linked with human obesity. Ad-36 causes obesity in chickens, mice, rats, and monkeys and was present in 30% of obese humans and 11% of nonobese humans. In twins discordant for infection with Ad-36, the infected twins were heavier and fatter than their cotwins. The growing body of evidence demonstrating that viruses produce human obesity supports the concept that at least some of the worldwide epidemic of obesity in the past 25 years is due to viral infections. PMID- 17908525 TI - Participation bias and its impact on the assembly of a genetic specimen repository for a myocardial infarction cohort. AB - OBJECTIVE: To assess participation bias in the assembly of a specimen repository for genetic studies and to examine the association of participation with outcome within the Olmsted County myocardial infarction (MI) cohort. PARTICIPANTS AND METHODS: From January 1, 1979, to May 31, 2006, 3081 persons had MI in Olmsted County, MN. Face-to-face contact was used to recruit patients who were hospitalized for an acute event. Persons who had had an MI before establishment of this repository were contacted by mail. At initial contact, we sought consent to use blood samples for genetic studies. Persons who refused were contacted by mail and were asked to consent to the use of stored tissue samples. For deceased subjects, stored tissue was collected when available. RESULTS: Of the 3081 persons in the Olmsted County MI cohort, 1994 participated in the study; 1007 (50.5%) blood and 987 (49.5%) tissue specimens were provided. Participants were more likely to be younger men with hypertension, comorbidities, and non-ST segment elevation MI (all, P<.05). Participants who provided blood specimens were more likely to have non-ST-segment elevation MI and lower Killip class than those who provided tissue. After adjustment for age, sex, hypertension, ST-segment elevation, Killip class, and comorbidities, participation was not associated with outcome. Participants who provided blood specimens were less likely to have heart failure (hazard ratio, 0.49; 95% confidence interval, 0.40-0.59; P<.01) or to die (hazard ratio, 0.16; 95% confidence interval, 0.12-0.21; P<.01) than those who provided tissue. CONCLUSION: A variety of sources can be used to assemble community specimen repositories. Baseline characteristics differed between participants and nonparticipants and, among participants, by specimen source. Participants who provided blood specimens had better outcomes than those who provided tissue specimens. No survival advantage was observed for participants after combining blood and tissue specimens. PMID- 17908527 TI - Sexual activity and chronic heart failure. AB - Little has been published about sexual function in chronic heart failure (CHF) and knowledge among clinicians in this regard is sparse. To review data regarding sexual function and dysfunction in patients with CHF, 2 of the authors (S.A.M. and P.A.U.) independently conducted a literature search using the MEDLINE database. English-language articles and cited bibliographies published between January 1996 and November 2006 were reviewed. Search terms included heart failure or CHF or ventricular dysfunction or heart disease in conjunction with sexual activity, erectile dysfunction, impotence, or sex. Articles were selected for inclusion if they had a primary focus on CHF and sexual function or dysfunction. Critical reviews of the literature, observational studies using self-reported patient surveys, and prospective, blinded, randomized, placebo-controlled trials were included. Articles were not excluded on the basis of patient sample size but were excluded if the article concerned a broad aspect of cardiovascular disease rather than CHF. When properly screened and treated, most patients with CHF can safely engage in sexual activity and be treated for erectile dysfunction with sildenafil, provided that they do not have active ischemia and do not require treatment with nitrates. Clinicians should know the physiological requirements of sexual activity and the impact CHF has on sexual performance. Fear of a cardiac event during intercourse can interfere with patients' ability to perform and enjoy sex, and so it is important that the physician be able to counsel patients with CHF about sexual activity. PMID- 17908528 TI - Applying quality-of-life data formally and systematically into clinical practice. AB - The systematic integration of quality-of-life (QOL) assessment into the clinical setting, although deemed important, infrequently occurs. Barriers include the need for a practical approach perceived as useful and efficient by patients and clinicians and the inability of clinicians to readily identify the value of integrating QOL assessments into the clinical setting. We discuss the use of QOL data in patient care and review approaches used to integrate QOL assessment into the clinical setting. Additionally, we highlight select QOL measures that have been successfully applied in the clinical setting. These measures have been shown to identify key QOL issues, improve patient-clinician communications, and improve and enhance patient care. However, the work done to date requires continued development. Continued research is needed that provides information about benefits and addresses limitations of current approaches. PMID- 17908529 TI - Exploration of the value of health-related quality-of-life information from clinical research and into clinical practice. AB - Quality-of-life (QOL) instruments used in clinical research can provide important evidence to inform decisions about alternative treatments. This is particularly true when patients, such as those with cancer who are contemplating toxic chemotherapy, face tradeoffs between quantity of life and QOL or when the primary goal of therapy is to improve how patients feel. Surrogate measures (cardiac function, exercise capacity, bone density, tumor size) are inadequate substitutes for direct measurement of QOL. Quality-of-life measures will be most valuable when they comprehensively measure aspects of QOL that are both important to patients and likely to be influenced by therapy, when the QOL measurement instruments are valid (measuring what is intended) and responsive (able to detect all important changes, even if small), and when the results are readily interpretable (determining whether treatment-related changes are trivial, small but important, or large). Researchers are finding new, imaginative ways to help clinicians understand the magnitude of treatment impact on QOL. Additionally, QOL measures may be useful in clinical practice. Recent results from well-designed randomized controlled trials suggest that information on patient QOL provided to clinicians might, in some circumstances, result in benefits for these patients. Further investigation is warranted to confirm these observations and to define the particular combination of methods and settings most likely to yield important benefits. PMID- 17908530 TI - Precision of health-related quality-of-life data compared with other clinical measures. AB - To many clinicians, the assessment of health-related quality of life (HRQL) seems more art than science. This belief is due in part to the lack of formal training available to clinicians regarding HRQL measurement and interpretation. When HRQL is used systematically, it has been shown to improve patient-physician communication, clinical decision making, and satisfaction with care. Nevertheless, clinicians rarely use formal HRQL data in their practices. One major reason is unfamiliarity with the interpretation and potential utility of the data. This unfamiliarity causes a lack of appreciation for the reliability of data generated by formal HRQL assessment and a tendency to regard HRQL data as having insufficient precision for individual use. This article discusses HRQL in the larger context of health indicators and health outcome measurement and is targeted to the practicing clinician who has not had the opportunity to understand and use HRQL data. The concept and measurement of reliability are explained and applied to HRQL and common clinical measures simultaneously, and these results are compared with one another. By offering a juxtaposition of common medical measurements and their associated error with HRQL measurement error, we note that HRQL instruments are comparable with commonly used clinical data. We further discuss the necessary requirements for clinicians to adopt formal, routine HRQL assessment into their practices. PMID- 17908531 TI - Evaluation and management of childhood and adolescent obesity. AB - The prevalence of obesity in children and adolescents has increased dramatically in the past 3 decades. Childhood and adolescent obesity are associated with serious comorbidities including type 2 diabetes mellitus, hyperlipidemia, and hypertension. Most obese children and adolescents have no defined underlying endocrine or genetic syndrome. Evaluation of an obese child or adolescent involves a detailed personal and family history, physical examination, and selected laboratory evaluation. Lifestyle interventions and behavioral modification aimed at decreasing caloric intake and increasing caloric expenditure are essential to management of childhood and adolescent obesity. Surgical approaches have a role in management of morbid obesity and serious obesity-related comorbidities in adolescents. Further research is needed to evaluate the role of various dietary approaches and pharmacotherapy in the treatment of obesity in childhood and adolescence. PMID- 17908532 TI - 33-Year-old man with cough, fever, and weight loss. PMID- 17908533 TI - Central nervous system tumors. AB - Central nervous system tumors are relatively common in the United States, with more than 40,000 cases annually. Although more than half of these tumors are benign, they can cause substantial morbidity. Malignant primary brain tumors are the leading cause of death from solid tumors in children and the third leading cause of death from cancer in adolescents and adults aged 15 to 34 years. Common presenting symptoms include headache, seizures, and altered mental status. Whereas magnetic resonance imaging helps define the anatomic extent of tumor, biopsy is often required to confirm the diagnosis. Treatment depends on the histologic diagnosis. Benign tumors are usually curable with surgical resection or radiation therapy including stereotactic radiation; however, most patients with malignant brain tumors benefit from chemotherapy either at the time of initial diagnosis or at tumor recurrence. Metastases to the brain remain a frequent and morbid complication of solid tumors but are frequently controlled with surgery or radiation therapy. Unfortunately, the mortality rate from malignant brain tumors remains high, despite initial disease control. This article provides an overview of current diagnostic and treatment approaches for patients with primary and metastatic brain tumors. PMID- 17908534 TI - Intentional ingestion of ethanol-based hand sanitizer by a hospitalized patient with alcoholism. PMID- 17908535 TI - Association between priapism and concurrent use of risperidone and Ginkgo biloba. PMID- 17908536 TI - Bloodstream infection prevention practices. PMID- 17908537 TI - Enamel, dentine and pulp in talon primary maxillary central incisors: a histomorphometric study. AB - AIM: To examine whether a talon incisor has an abnormal shape due to excess of dental hard tissues or pulp. METHODS: Two bilateral primary maxillary central incisors with talon cusp and a supernumerary mesiodens were removed from a 6-year old boy. Histologically, 7 undecalcified cross sections (70 microm) were harvested from each talon tooth and from two regular primary central incisors of another child. Microradiographs of the sections were prepared and examined for enamel width, dentin width, pulp tissue area, and crown size (mesio-distal, labio palatal) using Image Analysis. RESULTS: No association to other developmental disorders was observed. The mean width of the enamel and dentin was similar in the talon and the regular incisors (difference 7.5% and 2.4%, respectively). The pulp area was greater in the talon teeth (37.4%). The latter had an effect on the increase in the mesio-distal and labio-palatal dimensions of the talon teeth (17.6% and 23.9%, respectively). CONCLUSIONS: The fact that the talon cusp has normal enamel and dentin layers with a substantial enlarged pulp tissue suggests that similar developmental anomalies occur during tooth morphodifferentiation as in taurodontism and dens evagintus of premolars. PMID- 17908538 TI - Prevalence of dental trauma children aged 1-3 years in Joao Pessoa (Brazil). AB - AIM: This was to determine the prevalence and distribution of traumatic injuries to primary anterior teeth in children from 1 to 3 years-old. METHODS: A cross sectional survey was carried out through clinical examination of the primary dentitions of preschool children enrolled in public nursery schools in Joao Pessoa (Brazil). A total of 293 boys and girls participated in the study. The children were clinically examined for signs of trauma according to Andreasen's classification. RESULTS: Traumatic injuries were identified in 10.2% of children. The largest percentage of injuries was demonstrated by 2-3 years-old, with no significant difference between boys and girls. The most common type of injury was enamel fracture and enamel-dentine fracture. The maxillary central incisor was the most vulnerable to injury, without differences between the right and left side. STATISTICS: The data were entered in the SPSS program, and the chi square test was used with a 5% significance level. CONCLUSION: The primary dentition was most affected by fracture of enamel, especially the maxillary central incisor teeth, in patients between 2 and 3 years of age. There is a need of providing adequate preventive and treatment care for preschool children. PMID- 17908539 TI - Unexpected early apical resorption of primary molars and canines. AB - AIM: This was to examine the resorption pattern of primary molars and canines in dentitions with advanced apical resorption even though the permanent successor had barely begun root formation and to verify the resorption process histologically. STUDY DESIGN: Panoramic or bite-wing radiographs were selected from a dentition archive of radiographs from 142 children with deviant resorption patterns. METHODS: There were 14 patients (10 boys, 4 girls) aged 6 years 2 months to 8 years 1 month selected. In 11 patients abnormal resorption occurred only in the roots (group I). In 3 patients resorption occurred in the roots as well as in the crown (group II). The degree of resorption was evaluated using Haavikko's five resorption stages, and the degree of development of the permanent successor was evaluated using Haavikko's ten formation stages [Haavikko, 1973]. For histologic study 7 teeth were decalcified, paraffin-embedded, sectioned and stained with hematoxylin and eosin and examined microscopically. RESULTS: Group I: in 5 patients severe root resorption occurred before crown formation of the permanent successor was completed. In 6 children the abnormal resorption pattern was less severe. Group II: in 3 subjects full resorption of the root complex was observed as well as partial resorption of the crown. A histological analysis confirmed the resorption process. CONCLUSION: This is the first study focussing on unexpected early resorption of primary molars and canines in 14 patients without agenesis. The study showed an abnormal resorption pattern of roots and crown of primary teeth before the permanent successor had barely begun root formation. This indicates that resorption of primary molars and canines may occur independently from eruptional processes in the succeeding permanent tooth. PMID- 17908540 TI - Levels of oral disease in a sample of children with disability; a study carried out prior to comprehensive dental treatment under general anaesthesia. AB - AIM: To audit the levels of oral disease in those children whose disability required general anaesthesia for comprehensive dental treatment. METHODS: An audit was conducted of oral disease levels in a sample of 51 children attending for treatment. RESULTS: The proportion of untreated decayed teeth was 72%, previously extracted 25% and restored 3%. Very high levels of debris and gingival bleeding indices were found. There were very low proportions of previous restorations and past treatment had comprised mainly extractions. CONCLUSIONS: There was a low restorative care index in both the primary and permanent dentitions indicating that high needs continue to exist in terms of comprehensive dental care for these Irish children. PMID- 17908541 TI - Ectopic eruption of first permanent molars: presenting features and associations. AB - AIM: To investigate presenting features of ectopically erupting first permanent molars and associations with other dental anomalies. STUDY DESIGN: Prospective convenience study. METHODS: 28 panoral radiographs were collected, over a 24 month period, of 7-11 year-old children with radiographic evidence of ectopic eruption of first permanent molars who presented to a Dental Teaching Hospital in the North of England. A further 20 radiographs were collected of matched patients with no evidence of ectopic molar eruption. All radiographs were analysed under standard conditions to record the distribution and type of ectopic eruption (if present). In addition, the presence of the following dental anomalies was noted: cleft lip and/or palate; supernumerary teeth; hypodontia, and infraocclusion of primary molars. Chi-squared analysis was performed to determine any significant differences in the frequency of these dental anomalies between ectopic molar and control groups. RESULTS: For patients with ectopic molar eruption, the majority demonstrated ectopic eruption of either one or two first permanent molars (32% and 57% of subjects respectively). There were a similar proportion of 'jumps' and 'holds'. 92% of these were maxillary teeth and there was equal left and right distribution. Interestingly, a positive record of ectopic eruption was only documented in the dental records of 35.7% of these subjects. Children with ectopic eruption were significantly more likely to have at least one additional dental anomaly than was the case for the control group (60.7% versus 25%). Notably, primary molar infraocclusion and cleft lip/palate were significantly more frequent in the ectopic group. CONCLUSIONS: This study, the first in a British population, has identified a significant association between ectopic eruption of first permanent molars and other dental anomalies. A multifactorial aetiology is thus supported and clinicians should be alert to the co-existence of ectopic eruption and other dental anomalies. PMID- 17908542 TI - Diagnostic and restorative behaviour in Dutch dentists' paediatric dental care. AB - AIM: This was to assess if Dutch dentists have comparable attitudes in providing diagnosis and consecutive dental treatment in children. Prevention, radiographic diagnostics and restorative care were compared in four consecutive age groups [<6 years old, 6-8, 9-11, 12-17]. METHODS: In a cross-sectional study calculations were made using the financial records of one of the largest Dutch health insurance companies. After Medical Ethics approval all dental records of 4,500 dentists over a period of three consecutive years were used to calculate correlations and odds ratios, using early treatment, oral diagnosis, preventive approach and the use of local analgesia as basic independent variables. RESULTS: Early diagnosis based on the use of radiographs in children younger than 6 years increased the likelihood of direct restorative care 2.8 times and the likelihood of restorative care in future age groups 2.2, 2.0 and 1.6 times respectively. Early radiographs increased the likelihood of diagnostics in consecutive older groups by 3.7, 3.4 and 1.9 times respectively. CONCLUSION: An early diagnostic approach based on radiographs in paediatric dental care seems to be indicative for dentists' pattern of dealing with older children. This child-centred approach is limited to a small number of dentists. PMID- 17908543 TI - Clinical evaluation of three fissure sealants: 24 month follow-up. AB - AIM: This was to compare 3 different materials for fissure sealing (FS) after clinical use during the 24 month period and to assess the use of flowable composite resins in combination with dentine adhesives as sealing materials. STUDY DESIGN: There were 41 patients aged from 7-17, for whom 100 permanent molars were sealed with 3 different sealing materials. METHODS: Group A, 33 teeth were sealed with the material Helioseal Clear Chroma (Vivadent, Schaan, Liechtenstein); Group B, 33 teeth were sealed with the material Teethmate F1 (Kuraray, Kurashiki, Japan, and in Group C, 34 teeth were sealed with the flowable composite Tetric Flow (Vivadent, Schaan, Liechtenstein). For the analysis of the degree of retention, the criteria used were: 0--complete retention, 1--loss of (1/3) of material, 2--loss of (2/3) of material, 3- complete loss of material. Clinical criteria for appearance of a new caries lesion included following values: Caries present (+) or caries absent (-). STATISTICS: Chi-Square test was used for statistical analysis of retention rates between groups. RESULTS: After the 24 month period, the highest retention rate of complete retention was established for the material Tetric Flow of 76.5%, Helioseal Clear Croma 66.7%, and Teethmate F1 60.6%. The incidence of caries at Helioseal Clear Croma was 6.1%, for Teethmate F1 3%, with no statistically significant difference after the 24 months period (p=0.656). In the group of teeth sealed with Tetric Flow, there was not a single case of caries noted. CONCLUSIONS: The use of flowable composites as sealing materials is equal to other materials for fissure sealing. PMID- 17908544 TI - Knowledge of paediatric dentistry by recently-qualified general dentists: a pilot study. AB - AIM: To determine an overview of opinions and knowledge of paediatric dentistry amongst a group of recently-qualified dentists working within the Scottish Hospital Dental Service. STUDY DESIGN: A prospective questionnaire-based survey undertaken at Dundee Dental Hospital and School, NHS Tayside, Scotland. The survey evaluated dentists' opinion of managing the child patient and the location of dental treatment as well as knowledge of trauma, minor oral surgery, developmental disorders, mixed dentition problems and joint paediatric/orthodontic scenarios. RESULTS: Data were available for 15 dentists (M: 7; F: 8), median age 25.00 (inter-quartile range 25.40, 27.20) years. Of the respondents, 73.3% enjoyed treating children, although concerns regarding the management of both dental trauma and paediatric minor oral surgery cases were noted by 93.3% and 100% of dentists respectively. Concerning the location of treatment for the child patient, 66.7% believed that this should be 'shared-care' between the general dental practitioners (GDP) and Community Dental Service (CDS), whilst 20.0% and 13.3% observed that this should be undertaken within the CDS and GDP respectively. None of the dentists felt that treatment should be undertaken either privately or within the hospital services. Overall, dentists were best able to answer questions relating to mixed dentition and paediatric/orthodontic problems and least able to do so for minor oral surgical conditions. CONCLUSION: This pilot study offers baseline data regarding both opinions and also knowledge of paediatric dentistry amongst a group of recently qualified dentists. Further educational opportunities in this discipline at the postgraduate level may be beneficial. PMID- 17908545 TI - Assessment of two curing systems in a self-etching primer/adhesive sealant: a preliminary study for a clinical trial. AB - AIM: This was to assess the clinical performance of two different polymerization regimens of a non-rinse conditioning self/etching adhesive/sealant system (Adper Prompt-L-Pop, 3M ESPE) placed in recently erupted first permanent molars in two paediatric dental practices. METHODS: A total of 40 healthy 5 to 8 year old patients (20 from each practice) presenting at least two caries-free recently erupted first permanent molars participated in this preliminary test. A total of 128 molars were fissure sealed (FS) and cured with two different curing regimens. Group 1 (64 teeth) used Adper L-Pop + Clinpro as a one-step cure regimen. Group 2 (64 teeth) used Adper L-Pop + Clinpro with a two-step cure regimen using cotton roll isolation. Sealants were evaluated 6 to 12 months after placement. RESULTS: No differences were found in the ratings between the two polymerization regimens. In Group 1, 28 molars recorded FS fully retained rated A, 29 were rated B (partially missing) and 7 sealants were completely lost (C). In Group 2, 30 molars were rated A, another 30 scored B and 4 molars were totally lost (C). CONCLUSION: The poor performance of both FS polymerization regimens of the non rinse conditioning self/etching adhesive/sealant system (Adper Prompt-L-Pop, 3M ESPE) placed in recently erupted first permanent molars in the present test does not justify its use in young children. PMID- 17908551 TI - Silencing insulin resistance through SIRT1. AB - Insulin resistance is an important risk factor for the development of type 2 diabetes and the metabolic syndrome. A new study in this issue of Cell Metabolism (Sun et al., 2007) shows that SIRT1, a mammalian sirtuin homolog and histone deacetylase, can ameliorate insulin resistance by silencing expression of protein tyrosine phosphatase 1B, a major negative regulator of insulin action. PMID- 17908552 TI - Does FASing out new fat in the hypothalamus make you slim? AB - Fatty acid synthase (FAS) is a key enzyme for lipogenesis. A recent study (Chakravarthy et al., 2007) shows that the targeted deletion of FAS in hypothalamic neurons produces hypophagic, lean mice and suggests that PPARalpha signaling is an important mediator of the effects of FAS on energy homeostasis. PMID- 17908553 TI - Neuronal glucose sensing: still a physiological orphan? AB - Although hypothalamic glucose sensing is a long-established phenomenon, its physiological role remains unclear. New studies (Parton et al., 2007; Claret et al., 2007) disrupting glucose sensing in pro-opiomelanocortin neurons via differing methods have yielded disparate energy and glucose homeostasis phenotypes, suggesting that neuronal glucose sensing is not critical for these processes. PMID- 17908554 TI - Estrogen and bone: osteoclasts take center stage. AB - Loss of estrogen at menopause causes osteoporosis in many women, but estrogen's relevant cellular target in this process has remained unclear. In a recent study in Cell, Kato and colleagues (Nakamura et al., 2007) selectively ablate estrogen receptor alpha in osteoclasts and demonstrate that estrogen directly induces osteoclast apoptosis. PMID- 17908555 TI - Diabetic larvae and obese flies-emerging studies of metabolism in Drosophila. AB - The past few years have seen a shift in the use of Drosophila, from studies of growth and development toward genetic characterization of carbohydrate, sterol, and lipid metabolism. This research, reviewed below, establishes a new foundation for using this simple genetic model system to define the basic regulatory mechanisms that underlie metabolic homeostasis and holds the promise of providing new insights into the causes and treatments of critical human disorders such as diabetes and obesity. PMID- 17908556 TI - Foxa2 controls vesicle docking and insulin secretion in mature Beta cells. AB - The winged-helix transcription factor Foxa2 regulates Pdx1 gene expression and fetal endocrine pancreas development. We show here by inducible gene ablation that Foxa2 inactivation in mature beta cells induces hyperinsulinemic hypoglycemia in Foxa2(loxP/loxP),Pdx1-CreERT2 adult mice. Mutant beta cells exhibited a markedly increased pool of docked insulin granules, some of which were engaged in sequential or compound exocytosis, consistent with increased first-phase glucose-stimulated insulin secretion. Expression of multiple genes involved in vesicular trafficking, membrane targeting, and fuel-secretion pathways is dependent on Foxa2. In addition, impaired cytosolic Ca(2+) oscillations and elevated intracellular cyclic AMP production accompanied this secretory defect and were likely contributors to the sensitization of the exocytotic machinery. Thus, in the absence of Foxa2, alterations in intracellular second-messenger signaling redistribute the insulin granules into the readily releasable pool. We conclude that Foxa2 is required for both fetal pancreas development and the function of mature beta cells. PMID- 17908557 TI - Glucose restriction extends Caenorhabditis elegans life span by inducing mitochondrial respiration and increasing oxidative stress. AB - Increasing cellular glucose uptake is a fundamental concept in treatment of type 2 diabetes, whereas nutritive calorie restriction increases life expectancy. We show here that increased glucose availability decreases Caenorhabditis elegans life span, while impaired glucose metabolism extends life expectancy by inducing mitochondrial respiration. The histone deacetylase Sir2.1 is found here to be dispensable for this phenotype, whereas disruption of aak-2, a homolog of AMP dependent kinase (AMPK), abolishes extension of life span due to impaired glycolysis. Reduced glucose availability promotes formation of reactive oxygen species (ROS), induces catalase activity, and increases oxidative stress resistance and survival rates, altogether providing direct evidence for a hitherto hypothetical concept named mitochondrial hormesis or "mitohormesis." Accordingly, treatment of nematodes with different antioxidants and vitamins prevents extension of life span. In summary, these data indicate that glucose restriction promotes mitochondrial metabolism, causing increased ROS formation and cumulating in hormetic extension of life span, questioning current treatments of type 2 diabetes as well as the widespread use of antioxidant supplements. PMID- 17908558 TI - A conserved role for phosphatidylinositol 3-kinase but not Akt signaling in mitochondrial adaptations that accompany physiological cardiac hypertrophy. AB - Physiological cardiac hypertrophy is associated with mitochondrial adaptations that are characterized by activation of PGC-1alpha and increased fatty acid oxidative (FAO) capacity. It is widely accepted that phosphatidylinositol 3 kinase (PI3K) signaling to Akt1 is required for physiological cardiac growth. However, the signaling pathways that coordinate physiological hypertrophy and metabolic remodeling are incompletely understood. We show here that activation of PI3K is sufficient to increase myocardial FAO capacity and that inhibition of PI3K signaling prevents mitochondrial adaptations in response to physiological hypertrophic stimuli despite increased expression of PGC-1alpha. We also show that activation of the downstream kinase Akt is not required for the mitochondrial adaptations that are secondary to PI3K activation. Thus, in physiological cardiac growth, PI3K is an integrator of cellular growth and metabolic remodeling. Although PI3K signaling to Akt1 is required for cellular growth, Akt-independent pathways mediate the accompanying mitochondrial adaptations. PMID- 17908559 TI - SIRT1 improves insulin sensitivity under insulin-resistant conditions by repressing PTP1B. AB - Insulin resistance is often characterized as the most critical factor contributing to the development of type 2 diabetes. SIRT1 has been reported to be involved in the processes of glucose metabolism and insulin secretion. However, whether SIRT1 is directly involved in insulin sensitivity is still largely unknown. Here we show that SIRT1 is downregulated in insulin-resistant cells and tissues and that knockdown or inhibition of SIRT1 induces insulin resistance. Furthermore, increased expression of SIRT1 improved insulin sensitivity, especially under insulin-resistant conditions. Similarly, resveratrol, a SIRT1 activator, enhanced insulin sensitivity in vitro in a SIRT1-dependent manner and attenuated high-fat-diet-induced insulin resistance in vivo at a dose of 2.5 mg/kg/day. Further studies demonstrated that the effect of SIRT1 on insulin resistance is mediated by repressing PTP1B transcription at the chromatin level. Taken together, the finding that SIRT1 improves insulin sensitivity has implications toward resolving insulin resistance and type 2 diabetes. PMID- 17908560 TI - Inhibition of PKCepsilon improves glucose-stimulated insulin secretion and reduces insulin clearance. AB - In type 2 diabetes, pancreatic beta cells fail to secrete sufficient insulin to overcome peripheral insulin resistance. Intracellular lipid accumulation contributes to beta cell failure through poorly defined mechanisms. Here we report a role for the lipid-regulated protein kinase C isoform PKCepsilon in beta cell dysfunction. Deletion of PKCepsilon augmented insulin secretion and prevented glucose intolerance in fat-fed mice. Importantly, a PKCepsilon inhibitory peptide improved insulin availability and glucose tolerance in db/db mice with preexisting diabetes. Functional ablation of PKCepsilon selectively enhanced insulin release ex vivo from diabetic or lipid-pretreated islets and optimized the glucose-regulated lipid partitioning that amplifies the secretory response. Independently, PKCepsilon deletion also augmented insulin availability by reducing both whole-body insulin clearance and insulin uptake by hepatocytes. Our findings implicate PKCepsilon in the etiology of beta cell dysfunction and highlight that enhancement of insulin availability, through separate effects on liver and beta cells, provides a rationale for inhibiting PKCepsilon to treat type 2 diabetes. PMID- 17908561 TI - Glycogen synthase kinase 3alpha-specific regulation of murine hepatic glycogen metabolism. AB - Glycogen synthase kinase 3 comprises two isoforms (GSK-3alpha and GSK-3beta) that are implicated in type II diabetes, neurodegeneration, and cancer. GSK-3 activity is elevated in human and rodent models of diabetes, and various GSK-3 inhibitors improve glucose tolerance and insulin sensitivity in rodent models of obesity and diabetes. Here, we report the generation of mice lacking GSK-3alpha. Unlike GSK 3beta mutants, which die before birth, GSK-3alpha knockout (GSK-3alpha KO) animals are viable but display enhanced glucose and insulin sensitivity accompanied by reduced fat mass. Fasted and glucose-stimulated hepatic glycogen content was enhanced in GSK-3alpha KO mice, whereas muscle glycogen was unaltered. Insulin-stimulated protein kinase B (PKB/Akt) and GSK-3beta phosphorylation was higher in GSK-3alpha KO livers compared to wild-type littermates, and IRS-1 expression was markedly increased. We conclude that GSK-3 isoforms exhibit tissue-specific physiological functions and that GSK-3alpha KO mice are insulin sensitive, reinforcing the potential of GSK-3 as a therapeutic target for type II diabetes. PMID- 17908562 TI - Comparison of a functional restoration program with active individual physical therapy for patients with chronic low back pain: a randomized controlled trial. AB - OBJECTIVE: To compare the short-term outcomes of active individual therapy (AIT) with those of a functional restoration program (FRP). DESIGN: Prospective randomized controlled study. SETTING: Two rehabilitation centers and private ambulatory physiotherapy facilities. PARTICIPANTS: One hundred thirty-two adults with chronic low back pain. Fifty-one percent of patients on sick leave or out of work (mean duration, 180d in the 2y before treatment). INTERVENTIONS: For 5 weeks, FRP (at 25h/wk) or AIT (at 3h/wk). MAIN OUTCOME MEASURES: Trunk flexibility, back flexor, and extensor endurance (Ito and Sorensen tests), general endurance, pain intensity, Dallas Pain Questionnaire (DPQ) scores, daily activities, anxiety depression, social interest, and work and leisure activities, and self-reported improvement (work ability, resumption of sport and leisure activities). RESULTS: All outcome measures improved after treatment except endurance in AIT. There was no between-group difference for pain intensity or DPQ daily activities or work and leisure activities scores. Better results were observed in FRP for all other outcome measures. There was a significant effect of treatment and the initial value for the gain of the Sorensen score with a treatment or initial value interaction; a significant effect of treatment and initial value on the gains of Ito, endurance, and DPQ social interest and anxiety depression scores, with no treatment or initial value interaction; and a significant effect of initial value but not treatment for the gains of DPQ daily activities and work and leisure activities scores. CONCLUSIONS: Low-cost ambulatory AIT is effective. The main advantage of FRP is improved endurance. We speculate that this may be linked to better self-reported work ability and more frequent resumption of sports and leisure activities. PMID- 17908563 TI - Dual-task exercise improves walking ability in chronic stroke: a randomized controlled trial. AB - OBJECTIVE: To examine the effectiveness of a dual-task-based exercise program on walking ability in subjects with chronic stroke. DESIGN: Single-blind randomized controlled trial. SETTING: General community. PARTICIPANTS: Twenty-five subjects with chronic stroke who were at least limited community ambulatory subjects (a minimum gait velocity, 58cm/s). INTERVENTIONS: Participants were randomized into a control group (n=12) or experimental group (n=13). Subjects in the control group did not receive any rehabilitation training. Subjects in the experimental group underwent a 4-week ball exercise program. MAIN OUTCOME MEASURES: Gait performance was measured under single task (preferred walking) and tray-carrying task. Gait parameters of interest were walking speed, cadence, stride time, stride length, and temporal symmetry index. RESULTS: The experimental group showed significant improvement in all selected gait measures except for temporal symmetry index under both task conditions. In the control group, there were no significant changes over the 4-week period for all selected measures. There was a significant difference between groups for all selected gait variables except for temporal symmetry index under both task conditions. CONCLUSIONS: The dual-task based exercise program is feasible and beneficial for improving walking ability in subjects with chronic stroke. PMID- 17908564 TI - The effect of hippotherapy on spasticity and on mental well-being of persons with spinal cord injury. AB - OBJECTIVES: To determine the effect of hippotherapy on spasticity and on mental well-being of persons with spinal cord injury (SCI), and to compare it with the effects of other interventions. DESIGN: Crossover trial with 4 conditions. SETTING: Swiss paraplegic center. PARTICIPANTS: A volunteer sample of 12 people with spastic SCI (American Spinal Injury Association grade A or B). INTERVENTIONS: Hippotherapy, sitting astride a Bobath roll, and sitting on a stool with rocking seat. Each session lasted 25 minutes and was conducted twice weekly for 4 weeks; the control condition was spasticity measurement without intervention. MAIN OUTCOME MEASURES: Clinical rating by a blinded examiner of movement-provoked muscle resistance, using the Ashworth Scale; self-rating of spasticity by subjects on a visual analog scale (VAS); and mental well-being evaluated with the self-rated well-being scale Befindlichkeits-Skala of von Zerssen. Assessments were performed immediately after intervention sessions (short-term effect); data from the assessments were analyzed 3 to 4 days after the sessions to calculate the long-term effect. RESULTS: By analyzing the clinically rated spasticity, only the effect of hippotherapy reached significance compared with the control condition (without intervention); median differences in the Ashworth scores' sum before and after hippotherapy sessions ranged between 8.0 and +0.5. There was a significant difference between the spasticity-reducing effect of hippotherapy and the other 2 interventions in self-rated spasticity by VAS; median differences of the VAS before and after hippotherapy sessions ranged between -4.6 and +0.05cm. There were no long-term effects on spasticity. Immediate improvements in the subjects' mental well-being were detected only after hippotherapy (P=.048). CONCLUSIONS: Hippotherapy is more efficient than sitting astride a Bobath roll or on a rocking seat in reducing spasticity temporarily. Hippotherapy had a positive short-term effect on subjects' mental well-being. PMID- 17908566 TI - Musculoskeletal disorders in referrals for suspected cervical radiculopathy. AB - OBJECTIVES: To determine (1) the prevalence of selected common musculoskeletal disorders in patients referred for electrodiagnosis when cervical radiculopathy is suspected and (2) whether these findings predict electrodiagnostic study outcome. DESIGN: Prospective study. SETTING: Electrodiagnostic laboratories in departments of physical medicine and rehabilitation at 5 participating institutions. PARTICIPANTS: A total of 191 subjects undergoing electrodiagnostic evaluations for upper-limb symptoms when cervical radiculopathy was suspected. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Prevalence of certain musculoskeletal disorders (myofascial pain, shoulder impingement, lateral epicondylitis, de Quervain's tenosynovitis) and outcomes of electrodiagnostic testing (normal study, cervical radiculopathy, or another electrodiagnostically confirmed diagnosis). RESULTS: The total prevalence of musculoskeletal disorders was 42%. The prevalence in those with a normal study was 69%, compared with 29% in those with cervical radiculopathy (P<.001) and 45% in those with another diagnosis (P=.02). CONCLUSIONS: Musculoskeletal disorders are common in patients with suspected cervical radiculopathy. Although the presence of certain musculoskeletal disorders makes having a normal electrodiagnostic evaluation significantly more likely, the high prevalence among both patients with normal studies and those with radiculopathy and other disorders limits the usefulness of this information in precisely predicting study outcome. The presence of musculoskeletal disorders should not preclude electrodiagnostic testing when otherwise indicated. PMID- 17908565 TI - Does the presence of a specialized rehabilitation unit in a Veterans Affairs facility impact referral for rehabilitative care after a lower-extremity amputation? AB - OBJECTIVE: To determine if the presence of specialized rehabilitation units (SRUs) within Veterans Affairs medical centers (VAMC) influences access to rehabilitation services. DESIGN: Retrospective cohort analysis. SETTING: Two types of VAMCs: those with and without SRUs. PARTICIPANTS: Veterans with lower extremity amputations discharged from VAMCs between October 1, 2002, and September 30, 2003. There were a total of 2375 veterans with amputations: 99% were men; and 60% had transtibial, 40% had transfemoral, and less than 1% had hip disarticulation amputations. Nine hundred sixty-six patients (41%) were seen at a VAMC with an SRU. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Level of service provided expressed as: no evidence of rehabilitation during the hospitalization, generalized rehabilitation through consultation only, or admission to an SRU. RESULTS: There were no differences between patients treated at facilities with SRUs and those treated in a facility without SRU beds with respect to age, sex, marital status, source of hospital admission, or level of amputation (all P<.05). Patients with lower initial FIM instrument scores were more likely to be treated in facilities with SRUs, and to have longer lengths of acute hospitalization (P<.01). Patients at facilities with an SRU compared with those without an SRU had comparable likelihoods of being seen for an initial rehabilitation consultation (75% vs 74%, P=.56), but were more likely to be admitted for high intensity specialty rehabilitation services (26% vs 11%, P<.01). CONCLUSIONS: Although the majority of patients were seen in consultation, structural differences in service availability among clinically similar populations appear to be causing access disparities to specialized rehabilitation among amputees in the VAMC setting. The implication of these differences with regard to patient outcomes will need to be determined. PMID- 17908567 TI - The measurement of disability-related stress in wheelchair users. AB - OBJECTIVE: To measure disability-related stress through the development of the Physical Disability Stress Scale (PDSS) for wheelchair users. DESIGN: Cross sectional. SETTING: General community. PARTICIPANTS: Sample of 119 wheelchair users with an acquired physical disability. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: General Health Questionnaire-28 (GHQ) and the World Health Organization Quality of Life (WHOQOL-BREF) (Australian version). RESULTS: Factor analysis of PDSS items revealed 4 main factors of disability-related stress: access accounted for 33.7% of the variance, physical for 8.4% of the variance, social for 7.9% of the variance, and burden of care for 7.2% of the variance. Internal consistencies for the 4 factors were within acceptable ranges (alpha range, .78-.83). Concurrent validity was shown with the PDSS factors predicting 7% to 23% of the variance in GHQ subscales and total score and 12% to 31% of the WHOQOL-BREF subscales. Participants scoring in the GHQ psychiatric group showed significantly higher stress levels on the physical, social, and burden of care factors of the PDSS compared with the GHQ nonpsychiatric group. CONCLUSIONS: The results suggest the PDSS factors are valid measures of disability-related stress with potential for clinical and research applications. Confirmatory factor analyses with larger sample sizes of wheelchair users are required to establish consistency in the measurement of disability-related stress. PMID- 17908568 TI - Mobility assistive device utilization in a prospective study of patients with first-ever stroke. AB - OBJECTIVE: To estimate the extent to which clinical and functional features of stroke were related to the use of mobility assistive technology devices. DESIGN: Longitudinal study of quality of life after stroke. SETTING: Hospitals, rehabilitation centers, and universities in Ontario and Quebec. PARTICIPANTS: Subjects (N=316) with confirmed initial stroke were included in this analysis. Fifty-eight percent of the overall sample were men (n=184). The mean age of this sample at the time of the stroke +/- standard deviation was 65.3+/-15.3 years (range, 19-96y). One hundred thirty-five patients received a mobility assistive device poststroke, and 181 did not. INTERVENTION: Assistive devices for mobility (canes, walkers, wheelchairs). MAIN OUTCOME MEASURES: Assistive device use and mobility capacity. RESULTS: Mobility device nonusers were less physically disabled than device users on a variety of measures. Poor physical functioning but good cognition were reliably associated with mobility device use. Use of multiple mobility assistive devices was more often associated with poorer physical functioning than was single device use. For single device users, wheelchair use was predicted by cognition, functional independence, and stroke recovery. Cane users, compared with walker users, had better mobility and were less physically impaired by stroke. CONCLUSIONS: Patients were well matched to device type based on their mobility capacity. The findings of this study suggest that assistive device prescription-outcome relationships in stroke can be effectively and meaningfully modeled. PMID- 17908569 TI - Acupuncture for chronic shoulder pain in persons with spinal cord injury: a small scale clinical trial. AB - OBJECTIVE: To determine the efficacy of acupuncture in the treatment of chronic musculoskeletal shoulder pain in subjects with spinal cord injury (SCI). DESIGN: Randomized, double blind (participants, evaluator), placebo (invasive sham) controlled trial. SETTING: Clinical research center. PARTICIPANTS: Seventeen manual wheelchair-using subjects with chronic SCI and chronic musculoskeletal shoulder pain. INTERVENTIONS: Participants were randomly assigned to receive 10 treatments of either acupuncture or invasive sham acupuncture (light needling of nonacupuncture points). MAIN OUTCOME MEASURE: Changes in shoulder pain intensity were measured using the Wheelchair User's Shoulder Pain Index. RESULTS: Shoulder pain decreased significantly over time in both the acupuncture and the sham acupuncture groups (P=.005), with decreases of 66% and 43%, respectively. There was no significant difference between the 2 groups (P=.364). There was, however, a medium effect size associated with the acupuncture treatment. CONCLUSIONS: There appears to be an analgesic effect or a powerful placebo effect associated with both acupuncture and sham acupuncture. There was a medium treatment effect associated with the acupuncture, which suggests that it may be superior to sham acupuncture. This observation, along with the limited power, indicates that a larger, more definitive randomized controlled trial using a similar design is warranted. PMID- 17908570 TI - Cognitive impairment in patients with traumatic brain injury and obstructive sleep apnea. AB - OBJECTIVE: To examine the impact of comorbid obstructive sleep apnea (OSA) on the cognitive functioning of traumatic brain injury (TBI) patients. DESIGN: A case control study. Neuropsychologic test performances of TBI patients with OSA were compared with those who did not have OSA. The diagnosis of OSA was based on standard criteria using nocturnal polysomnography. SETTING: Three academic medical centers with level I trauma centers, accredited sleep disorders centers, and rehabilitation medicine programs. PARTICIPANTS: Thirty-five TBI patients who were part of a project that assessed the effect of sleep disorders in a larger sample of consecutively recruited TBI patients. There were 19 patients with TBI and OSA. They were compared with 16 TBI patients without OSA who were comparable in terms of age, education, severity of injury (when available), time postinjury, and Glasgow Coma Scale scores (when available). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The Psychomotor Vigilance Test, Rey Complex Figure Test, Rey Auditory Verbal Learning Test, digit span test from the Wechsler Memory Scale Revised, and finger-tapping test. RESULTS: The TBI patients with OSA performed significantly worse than the non-sleep disordered TBI patients on verbal and visual delayed-recall measures. The groups performed comparably on motor, visual construction, and attention tests. The TBI patients with OSA made more attention lapses (reaction times >/=500ms), but showed comparable fastest and slowest reaction times on a measure of sustained attention. CONCLUSIONS: OSA is associated with more impairment of sustained attention and memory in TBI patients. It is possible that early identification and treatment of OSA may improve cognitive, and thus potentially functional, outcomes of TBI patients with this disease. PMID- 17908571 TI - Development of a French isometric strength normative database for adults using quantitative muscle testing. AB - OBJECTIVE: To establish a normative database for isometric strength measured by quantitative muscle testing (QMT) for a French adult population. DESIGN: Measurement of maximal voluntary isometric contraction. SETTING: Four clinical centers involved in neuromuscular disorders. PARTICIPANTS: A total of 315 healthy adults (147 men, 168 women) ages 20 to 80 years. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Isometric torque values of 14 muscle functions (13 bilaterally and neck). RESULTS: This study led to the development of a French isometric strength normative database for adults measured by QMT. For each muscle function, predictive regression models using age, sex, and weight are proposed. Some methodologic issues concerning strength measurement are discussed. CONCLUSIONS: This database can be used to compute relative deficits in muscle strength for 27 muscle functions and also to estimate composite scores for follow up of patients either during the natural history of their disease or during a therapeutic trial. PMID- 17908572 TI - Measurement of energy cost by the physiological cost index in walking after stroke. AB - OBJECTIVE: To compare the Physiological Cost Index (PCI) with direct measurement of oxygen consumption (Vo(2)) as an estimate of energy cost in persons with stroke and healthy subjects. DESIGN: Test-retest on separate days. A comparison of 2 methods of measurement. Measurements with and without an orthosis. SETTING: A university hospital. PARTICIPANTS: A convenience sample of 20 persons with hemiparesis more than 6 months after stroke and 16 healthy subjects, ages 30 to 63 years. INTERVENTIONS: Five minutes of treadmill walking at self-selected speeds while recording Vo(2) levels and heart rates. Additional data was recorded for 11 of the stroke subjects with and without an ankle-foot orthosis. MAIN OUTCOME MEASURES: Vo(2) and the PCI. RESULTS: No significant differences were found in the PCI or Vo(2) between test and retest. Both PCI and Vo(2) per distance were higher for the stroke subjects compared with healthy subjects. PCI showed a larger dispersion than Vo(2) between test and retest. The regression analysis for PCI showed that the model including age, sex, group assignment, and Vo(2) could explain 53% of the variation. The PCI did not show a significant difference in walking with or without an orthosis, whereas Vo(2) differed significantly. CONCLUSIONS: The PCI showed limited reliability and validity as a measure of energy cost after stroke due to the extensive variability between test and retest. PMID- 17908573 TI - Validity of the trunk impairment scale as a measure of trunk performance in people with Parkinson's disease. AB - OBJECTIVE: To evaluate construct validity of the Trunk Impairment Scale (TIS) as a measure of trunk performance in Parkinson's disease (PD). DESIGN: A cross sectional study of PD patients and healthy subjects. SETTING: University rehabilitation research unit. PARTICIPANTS: Twenty-six PD patients (Hoehn and Yahr stages 2-4) and 26 healthy subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The TIS and its subscales; static and dynamic sitting balance and trunk coordination. RESULTS: Compared with healthy controls, PD patients showed significantly lower scores on the total TIS, static sitting balance, and coordination subscale. Healthy subjects scored significantly better on the total TIS and coordination subscale compared with patients in the early stage of PD. Patients with PD in the early stage scored significantly higher for the total TIS as well as static and dynamic sitting balance in comparison with PD patients in a later stage. Forward stepwise multiple linear regression analysis showed that trunk impairment in PD patients was significantly related to a combination of older age and a higher score on part III of the Unified Parkinson's Disease Rating Scale, which assesses motor impairments. CONCLUSIONS: Early detection of trunk deficits and the significant relation with PD severity advocates further evaluation and use of the TIS in PD. PMID- 17908574 TI - The effect of a dual-task on obstacle crossing in healthy elderly and young adults. AB - OBJECTIVE: To investigate the effect of a dual-task on step initiation over an obstacle. DESIGN: Repeated-measures design between groups. SETTING: University laboratory. PARTICIPANTS: Ten healthy, community-dwelling elderly adults and 10 healthy young adults. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The effect of a reaction time task on ground reaction forces, toe clearance, and temporal events in stepping over an obstacle. RESULTS: Ground reaction forces of the swing limb before toe-off did not differ between the groups and was not affected by task difficulty. Stepping with a random stimulus after toe-off was the most difficult task, whereas stepping with a predictable stimulus before toe off was the easiest task. Reaction time and stepping time were greater and toe clearance was less for the elderly subjects. Both groups had a decrease in toe clearance and an increase in reaction time and stepping time as the task became more difficult. CONCLUSIONS: Our findings indicate that even healthy older adults may be at risk for falls in situations where they are engaged in concurrent tasks. The data support the inclusion of dual-task activities in fall prevention programs. PMID- 17908575 TI - Predictive validity and responsiveness of the functional ambulation category in hemiparetic patients after stroke. AB - OBJECTIVE: To determine the reliability, concurrent and predictive validity, and responsiveness of the Functional Ambulation Category (FAC) in hemiparetic patients after stroke. DESIGN: Prospective cohort. SETTING: An early rehabilitation center for patients with neurologic disorders. PARTICIPANTS: Fifty five nonambulatory patients after first-ever stroke, with duration of illness between 30 and 60 days, were included. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: FAC, Rivermead Mobility Index (RMI), walking velocity, step length, and six-minute walking test (6MWT) were assessed at the beginning, after 2 and 4 weeks of rehabilitation, and again 6 months later. After 6 months, community ambulation was also assessed. Test-retest and interrater reliability, concurrent, discriminant, and predictive validity and responsiveness of the FAC were calculated. RESULTS: Based on video examinations, high test-retest reliability (Cohen kappa=.950) and interrater reliability (kappa=.905) were found. FAC scores at the beginning and after 2 weeks, 3 weeks, and 6 months correlated highly with the RMI (Spearman rho=.686, rho=.787, rho=.825, rho=.893, respectively), distance walked in the 6MWT (rho=.949, rho=.937, rho=.931, rho=.906, respectively), walking velocity (rho=.952, rho=.939, rho=.902, rho=.901, respectively), and step length (rho=.952, rho=.932, rho=.896, rho=.877, respectively) at the same time points (all P<.001). The RMI, walking velocity, step length, and distance walked in the 6MWT differed for each FAC category (P<.001). After 4 weeks of rehabilitation, an FAC score of 4 or higher predicted community ambulation at 6 months with 100% sensitivity and 78% specificity. FAC scores changed significantly between the first 2 and second 2 weeks (Wilcoxon z=8.7, z=7.9, respectively; both P<.001) of the inpatient rehabilitation program. CONCLUSIONS: The FAC has excellent reliability, good concurrent and predictive validity, and good responsiveness in patients with hemiparesis after stroke. PMID- 17908576 TI - Fast-track programming and rehabilitation model: a novel approach to postoperative deep brain stimulation patient care. AB - OBJECTIVE: To propose a new model of integrated, multidisciplinary postoperative care of the patients with deep brain stimulation (DBS). DESIGN: Observational cohort study with follow-up at 3 months and 1 year. SETTING: Academic medical center movement disorder clinic. PARTICIPANTS: Seventy-three consecutive patients with medically refractory Parkinson's disease underwent bilateral DBS. Patients were then transferred directly to an inpatient rehabilitation facility. INTERVENTION: DBS and inpatient programming and rehabilitation. Simultaneous programming and rehabilitation was carried out by a multidisciplinary team. MAIN OUTCOME MEASURES: The FIM instrument, Unified Parkinson Disease Rating Scale (UPDRS), and levodopa dosage. RESULTS: The average rehabilitation stay was 17.3 days, with a mean of 6.2 stimulator adjustments during that time. FIM scores improved from 62.1 (admission) to 98.5 (discharge), an average improvement of 36.4 (58.6%). Average UPDRS scores improved from 52.5 (preoperative off) and 30.1 (preoperative on) to 20.4 (3mo postoperative on-medication, on-stimulation), a 32.2% improvement from the preoperative on score. Levodopa dosages decreased by an average of 48.3% (all P<.001). CONCLUSIONS: We describe our fast-track protocol, which allows for rapid DBS programming and tapering of Parkinson's medications. It also provides for treatment of concomitant medical and psychologic problems and optimized physical performance. PMID- 17908577 TI - Coordination between reaching and grasping in patients with hemiparesis and healthy subjects. AB - OBJECTIVE: To investigate the coordination of reach-to-grasp components in hemiparetic and healthy subjects. DESIGN: Split-plot repeated-measures design with 3 factors (group, object size, movement speed). SETTING: Movement laboratory. PARTICIPANTS: Twelve hemiparetic and 12 age-matched healthy subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: We used motion analysis to collect information on the kinematic variables of movement duration, peak velocity, peak deceleration, and maximum aperture, and the time of peak velocity, peak deceleration, and maximum aperture expressed as a percentage of movement duration during 32 reaching movements by each subject. We examined the coordination between the 2 components in 2 ways. First, we investigated the correlation between time of hand opening and start of hand transport, and between time of maximum aperture and time of peak deceleration. Second, we compared movements at preferred and fast speeds (manipulation of transport component) and to 2 different-sized cups (manipulation of grasp component). RESULTS: Both groups demonstrated a temporal coupling between grasp and transport components at the start of the reach and at the time of maximum aperture. Both groups increased the aperture of grasp for larger cups and increased the maximum grip aperture, and had a shorter deceleration phase for faster movements. The deceleration phase of the hemiparetic patients was longer than that of the healthy subjects, however, and the components were not as tightly coupled. CONCLUSIONS: The hemiparetic patients, who had a moderate amount of functional recovery, were similar to healthy subjects in their ability to control reach-to-grasp components. Their performance was not as skilled, however. PMID- 17908578 TI - Effects of robot-aided bilateral force-induced isokinetic arm training combined with conventional rehabilitation on arm motor function in patients with chronic stroke. AB - OBJECTIVE: To analyze the effects of conventional rehabilitation combined with bilateral force-induced isokinetic arm movement training on paretic upper-limb motor recovery in patients with chronic stroke. DESIGN: Single-cohort, pre- and postretention design. SETTING: Rehabilitation department at a medical university. PARTICIPANTS: Twenty subjects who had unilateral strokes at least 6 months before enrolling in the study. INTERVENTION: A training program (40min/session, 3 sessions/wk for 8wk) consisting of 10 minutes of conventional rehabilitation and 30 minutes of robot-aided, bilateral force-induced, isokinetic arm movement training to improve paretic upper-limb motor function. MAIN OUTCOME MEASURES: The interval of pretest, post-test, and retention test was set at 8 weeks. Clinical arm motor function (Fugl-Meyer Assessment [FMA], upper-limb motor function, Frenchay Arm Test, Modified Ashworth Scale), paretic upper-limb strength (grip strength, arm push and pull strength), and reaching kinematics analysis (peak velocity, percentage of time to peak velocity, movement time, normalized jerk score) were used as outcome measures. RESULTS: After comparing the sets of scores, we found that the post-test and retention test in arm motor function significantly improved in terms of grip (P=.009), push (P=.001), and pull (P=.001) strengths, and FMA upper-limb scale (P<.001). Reaching kinematics significantly improved in terms of movement time (P=.015), peak velocity (P=.035), percentage of time to peak velocity (P=.004), and normalized jerk score (P=.008). Improvement in reaching ability was not sustained in the retention test. CONCLUSIONS: Preliminary results showed that conventional rehabilitation combined with robot-aided, bilateral force-induced, isokinetic arm training might enhance the recovery of strength and motor control ability in the paretic upper limb of patients with chronic stroke. PMID- 17908579 TI - Hyperbaric gaseous cryotherapy: effects on skin temperature and systemic vasoconstriction. AB - OBJECTIVE: To compare skin-surface cooling caused by the application of an ice bag (15min) and the projection of carbon dioxide microcristals (2min) under high pressure (75 bar) and low temperature (-78 degrees C), a modality called hyperbaric gaseous cryotherapy. DESIGN: Randomized controlled trial with repeated measure. SETTING: Laboratory experiment. PARTICIPANTS: Twelve healthy male subjects (mean +/- standard deviation, 22.9+/-1.8y). INTERVENTIONS: Ice bag and hyperbaric gaseous cryotherapy were randomly applied on the skin of the nondominant hand. MAIN OUTCOME MEASURE: Skin temperature of the cooled (dorsal and palmar sides) and contralateral (dorsal side) hands were continuously measured with thermistor surface-contact probes before, during, and after (30min) cooling. RESULTS: Hyperbaric gaseous cryotherapy projection induced a large decrease (P<.05) of the dorsal skin temperature of the cooled hand (from 32.5 degrees +/-0.5 degrees C to 7.3 degrees +/-0.8 degrees C) and a significant decrease of the skin temperature of the palmar side and of the contralateral hand. The skin temperature of the dorsal side of the cooled hand was decreased with an ice bag (from 32.5 degrees +/-0.6 degrees C to 13.9 degrees +/-0.7 degrees C, P<.05). However, the lowest temperature was significantly higher than during hyperbaric gaseous cryotherapy, and no significant changes in the other skin temperatures were observed. Rewarming was equal after the 2 modalities, highlighting a more rapid increase of the skin temperature after hyperbaric gaseous cryotherapy. CONCLUSIONS: Hyperbaric gaseous cryotherapy projection decreased the skin temperature of the cooled and contralateral hand, suggesting a systemic skin vasoconstriction response. On the other hand, the vascular responses triggered by ice pack cooling appeared limited and localized to the cooled area. PMID- 17908580 TI - Alternating frequencies of transcutaneous electric nerve stimulation: does it produce greater analgesic effects on mechanical and thermal pain thresholds? AB - OBJECTIVE: To determine whether alternating frequency transcutaneous electric nerve stimulation (TENS) at 2 and 100Hz (2/100Hz) has a more potent hypoalgesic effect than a fixed frequency at 2 or 100Hz in healthy participants. DESIGN: A single-blind randomized controlled trial with a convenience sample. SETTING: University physiotherapy department. PARTICIPANTS: Sixty-four healthy volunteers (32 men [mean age, 28.1+/-5.9y], 32 women [mean age, 27.7+/-5.6y]) were recruited and randomly divided into 4 groups. INTERVENTIONS: The 4 groups received TENS delivered at (1) 2Hz; (2) 100Hz; (3) 2/100Hz alternating frequency; and (4) no treatment (control group), respectively. Electric stimulation was applied over the anterior aspect of the dominant forearm for 30 minutes. MAIN OUTCOME MEASURES: Mechanical pain thresholds (MPTs) and heat pain thresholds (HPTs) were recorded before, during, and after TENS stimulation. The data were analyzed using linear mixed models, with group treated as a between-subject factor and time a within-subject factor. RESULTS: During and shortly after electric stimulation, HPT increased significantly in the alternating frequency stimulation group (P=.024). MPT increased significantly in both the 100Hz (P=.008) and the alternating frequency groups (P=.012), but the increase was substantially larger in the 100Hz group. CONCLUSIONS: Alternating frequency stimulation produced a greater elevation in the HPT, but a greater increase in the MPT was achieved using 100Hz stimulation. PMID- 17908581 TI - Irreversible spinal nerve injury from dorsal ramus radiofrequency neurotomy: a case report. AB - Radiofrequency neurotomy (RFN) of the medial branches of the dorsal rami is a successful method of treating facet joint pain. Documented serious complications are rare. We discuss the case of a 33-year-old woman with low back pain (LBP) who sustained a right L5 nerve root injury during RFN. The patient had several months of axial LBP after a motor vehicle collision. She had no relief after anti inflammatory medications, physical therapy, L5-S1 interlaminar epidural corticosteroid injections, and a right sacroiliac joint injection. She then received bilateral L3 and L4 medial branch and bilateral L5 dorsal ramus blocks with excellent temporary pain relief. Subsequently she underwent bilateral L3 and L4 medial branch and bilateral L5 dorsal ramus RFN. Afterward, she noticed new right leg pain and paresthesias extending throughout the L5 dermatome. Electromyography and magnetic resonance imaging were normal and she was diagnosed with a right L5 sensory radiculopathy. The right leg symptoms were unresponsive to multiple medications. After a successful trial with a spinal cord stimulator, she underwent permanent stimulator placement. Afterward, she had 90% relief of her right leg pain and discontinued all analgesics. Irreversible injury of nontarget nerves is a possible complication of RFN, and can be avoided by following proper procedural protocol. PMID- 17908583 TI - Acute bacterial sacroiliitis in an adult: a case report and review of the literature. AB - Bacterial septic sacroiliitis is an uncommon diagnosis that occurs most frequently in children and young adults. Nonspecific physical examination findings often make it difficult to diagnose the condition, thus delaying appropriate treatment. We review the case of middle-aged woman with sacroiliac joint (SIJ) pain after a torsional injury. Radiographic films showed the pelvis and left lower extremity to be normal. Despite anti-inflammatory medications, analgesics, a corticosteroid injection, and physical therapy, her pain persisted. Laboratory data showed an elevated erythrocyte sedimentation rate and C-reactive protein; otherwise, tests were normal, including negative blood cultures. Magnetic resonance imaging (MRI) revealed a left posteroinferior SIJ effusion and computed tomography (CT) showed an effusion and irregularity in the left SIJ. An SIJ biopsy revealed inflammation suggestive of osteomyelitis. After a course of intravenous antibiotics, the symptoms completely resolved, thus supporting our diagnosis of bacterial sacroiliitis. Repeat MRI and CT confirmed the complete resolution of the sacroiliitis. PMID- 17908582 TI - Interspinous ligament steroid injections for the management of Baastrup's disease: a case report. AB - Mitra R, Ghazi U, Kirpalani D, Cheng I. Interspinous ligament steroid injections for the management of Baastrup's disease: a case report. Baastrup's disease has been identified as a source of axial low back pain. There has been debate as to the etiology of pain in patients with Baastrup's disease. It has been theorized that the pain may originate from degenerative disk disease and spinal stenosis associated with the disease, whereas some have identified the neoarthrosis between joints and accompanying reactive eburnation as the source of pain. We present a simple case report of an 89-year-old woman with symptomatic Baastrup's disease. The patient underwent a fluoroscopically guided interspinous process injection of 20mg of triamcinolone acetate with local anesthetic. The patient remained pain free for 3 months. The neoarthrosis in Baastrup's disease may be the primary pain generator in cases of Baastrup's disease without significant central canal stenosis. PMID- 17908584 TI - Further validation of the Orientation and Cognitive Logs: their relationship to the Mini-Mental State Examination. AB - OBJECTIVE: To further evaluate the construct validity of bedside screening measures of orientation (Orientation Log [O-Log]) and cognition (Cognitive-Log [Cog-Log]) by examining the relationship between these measures and the Mini Mental State Examination (MMSE). DESIGN: Correlational analysis used to assess the degree of overlapping variance among the O-Log, Cog-Log, and MMSE. Qualitative item analysis used to assess strengths and weaknesses of the measures. SETTING: Inpatient rehabilitation center affiliated with a large university medical school. PARTICIPANTS: Participants were 45 inpatients receiving neurorehabilitation. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The O-Log, Cog-Log, and MMSE. RESULTS: The MMSE correlated significantly with both measures (O-Log, r=.65, P<.001; Cog-Log, r=.75, P<.001). The O-Log and C-Log were significantly related to each other (r=.75, P<.001). CONCLUSIONS: The results indicated good construct validity of the O-Log and Cog Log. These measures may be better suited for a population with moderate to severe brain injury in a rehabilitation setting, compared with the MMSE, because scales were developed to give partial credit based on partially correct answers. Further, the O-Log and C-Log do not have a written component, allowing administration for persons with hemiparesis. PMID- 17908585 TI - Ultrasound-guided blockade of the lateral femoral cutaneous nerve: technical description and review of 10 cases. AB - Blockade of the lateral femoral cutaneous nerve (LFCN) is performed for therapeutic management of meralgia paresthetica and as a regional anesthetic technique. The conventional technique is associated with high failure rates secondary to variable LFCN anatomy. We describe a technique for blockade of the LFCN using ultrasound guidance. A cross-sectional view of the LFCN was obtained by identifying the anterior superior iliac spine, then moving a 14-to-7MHz linear array ultrasound probe in a medial caudal direction until the nerve was encountered. The needle was advanced to the LFCN under ultrasound guidance via a lateral to medial approach. Injection using dynamic ultrasound demonstrated excellent perineural spread. Ten subjects underwent successful blockade of the LFCN with this technique. Five subjects were obese. Use of ultrasound for precise needle placement allowed low injection volumes to be utilized. Theref were no complications. Ultrasound guidance can facilitate blockade of the LFCN for diagnostic and therapeutic purposes and may be particularly beneficial with patients with challenging surface anatomic landmarks, or when low volume injections are desired. PMID- 17908586 TI - Proceedings of the 5th International Conference on Cervical Cancer. PMID- 17908588 TI - The clinical effectiveness of optical spectroscopy for the in vivo diagnosis of cervical intraepithelial neoplasia: where are we? AB - OBJECTIVE: In this review, we evaluate the diagnostic efficacy of optical spectroscopy technologies (fluorescence and reflectance spectroscopy) for the in vivo diagnosis of cervical neoplasia using both point probe and multispectral imaging approaches. METHODS: We searched electronic databases using the following terms: cervical cancer, cervical intraepithelial neoplasia, squamous intraepithelial lesion, and spectroscopy, fluorescence spectroscopy, or reflectance spectroscopy. We included studies that evaluated fluorescence and reflectance spectroscopy devices for in vivo diagnosis, compared those results with biopsy results, and reported on the sensitivity and specificity of the devices tested. RESULTS: Twenty-six studies, including seven phase II trials and one randomized clinical trial, met our acceptability criteria. We found several important differences across the studies including device approach (multispectral versus point probe), study population, disease classification system, and disease threshold. This heterogeneity prevented formal combination of sensitivity and specificity results. CONCLUSION: Optical spectroscopy has similar performance to colposcopy and may help localize lesions and therefore be an effective adjunct to colposcopy. Reports on the diagnostic accuracy of these devices should use common thresholds for the construction of receiver operating characteristic curves to enable comparisons with standard technologies and facilitate their adoption. Optical spectroscopy has also been identified for possible use as ASCUS triage and primary screening, yet neither has been sufficiently evaluated to warrant a conclusion as to their suitability in this role. PMID- 17908587 TI - Bayesian meta-analysis of Papanicolaou smear accuracy. AB - OBJECTIVE: To perform a Bayesian analysis of data from a previous meta-analysis of Papanicolaou (Pap) smear accuracy (Fahey et al. Am J Epidemiol 1995; 141:680 689) and compare the results. METHODS: We considered two Bayesian models for the same data set used in the Fahey et al. study. Model I was a beta-binomial model which considered the number of true positives and false negatives as independent binomial random variables with probability parameters beta (sensitivity) and alpha (one minus specificity), respectively. We assumed that beta and alpha are independent, each following a beta distribution with exponential priors. Model II considered sensitivity and specificity jointly through a bivariate normal distribution on the logits of the sensitivity and specificity. We performed sensitivity analysis to examine the effect of prior selection on the parameter estimates. RESULTS: We compared the estimates of average sensitivity and specificity from the Bayesian models with those from Fahey et al.'s summary receiver operating characteristics (SROC) approach. Model I produced results similar to those of the SROC approach. Model II produced point estimates higher than those of the SROC approach, although the credible intervals overlapped and were wider. Sensitivity analysis showed that the Bayesian models are somewhat sensitive to the variance of the prior distribution, but their point estimates are more robust than those of the SROC approach. CONCLUSIONS: The Bayesian approach has advantages over the SROC approach in that it accounts for between study variation and allows for estimating the sensitivity and specificity for a particular trial, taking into consideration the results of other trials, i.e., "borrowing strength" from other trials. PMID- 17908589 TI - Web 2.0, library 2.0, physician learning 2.0. PMID- 17908590 TI - Visual acuity as an outcome measure in clinical trials of retinal diseases. AB - PURPOSE: Visual acuity (VA) is the primary outcome measure in many studies involving eye diseases. A standard statistical approach for comparing a continuous measurement such as a VA letter score between 2 treatment groups is to perform a t test comparing the means. However, frequently a binary variable is created from the continuous VA letter score based on whether or not there has been a worsening (or gain) of > or =15 letters (equivalent to > or =3 lines), and a chi square or similar statistical test is performed to compare the proportions of success (or failure) between groups. The purpose of this article is to contrast these 2 approaches. METHODS: Clinical trial reports of retinal disorders were used to compare results using mean change in the VA letter score versus binary proportions created from the VA letter score. Additionally, analyses were performed using generated data to gain a perspective on the magnitude of differences that might be expected between the 2 methods. RESULTS: Studies from the literature showed that differences of 6% to 15% in > or =15-letter worsening corresponded to mean differences in letter scores between groups of 3.0 to 7.0 (approximately 0.6 to 1.4 lines). Analyses using generated data demonstrated that a mean improvement in the VA letter score of 5 corresponded to a doubling of the proportion of eyes with > or =15-letter improvement and a 28% relative reduction in the proportion of eyes with > or =15-letter worsening. CONCLUSIONS: How VA data should be analyzed in a clinical trial depends to large extent on the research question. The frequently used outcome of > or =15-letter change has several drawbacks, including loss of efficiency (need for a larger sample), misclassification of the outcome, and potential for a ceiling or floor effect. Therefore, for most clinical trials we believe that the primary outcome analysis should be a comparison of changes in the VA letter score, and created binary variables should be reported as secondary outcomes. This approach maximizes the information gained from the data and accommodates both improvement and worsening of acuity. PMID- 17908591 TI - A phase III study of subconjunctival human anti-transforming growth factor beta(2) monoclonal antibody (CAT-152) to prevent scarring after first-time trabeculectomy. AB - OBJECTIVE: To evaluate CAT-152 (lerdelimumab), a monoclonal antibody to transforming growth factor-beta2 (TGF-beta2), in preventing the progression of fibrosis in patients undergoing first-time trabeculectomy for primary open-angle (POAG) or chronic angle-closure glaucoma (CACG). DESIGN: Randomized, double masked, multicenter, placebo-controlled trial. PARTICIPANTS: Individuals with a diagnosis of POAG, CACG, pseudoexfoliative glaucoma (PEXG), or pigmentary glaucoma (PG), with a recorded intraocular pressure (IOP) of more than 21 mmHg, visual field or optic disc changes characteristic of glaucoma, and taking the maximum tolerated dose of medication. INTERVENTION: Patients received unilateral trabeculectomy with either 4 subconjunctival injections of CAT-152 (100 microg in 100 microl phosphate buffer) or 4 placebo injections, administered immediately before and on completion of trabeculectomy, and on the first day and at 1 week after surgery. Patients were followed up for 12 months after surgery. MAIN OUTCOME MEASURES: The primary outcome measure was treatment success in the study eye (unmedicated IOP of 6-16 mmHg inclusive), at the 6- and 12-month follow-up. Secondary outcome measures were the incidence of postoperative intervention with 5-fluorouracil (5-FU); incidence of surgical failure; time to surgical failure; and incidence of vascularity, microcysts, and encapsulation or demarcation of the bleb site. RESULTS: Of the 388 patients evaluated in the trial, 81% (n = 274) had either POAG or CACG, combined into a single set (POAG/CACG) analyzed by intent-to treat (ITT) criteria. Separate ITT analyses were carried out for all participants (+PEXG/PG group), with similar results. The treatment success rate was 60% in the CAT-152 group and 68% in the placebo group (P = 0.23). No statistically significant differences emerged in the secondary end points. Patients requiring 5 FU for postsurgical management were more likely to be treatment failures (P = 0.0003). Patients with a primary diagnosis of PG (n = 49) had a higher success rate than those with other diagnoses (P = 0.0077). Administration of CAT-152 was not associated with an increased incidence of adverse events. The immunogenicity of CAT-152 was very low. CONCLUSIONS: At the dose level and regimen studied, there was no difference between CAT-152 and placebo in preventing the failure of primary trabeculectomy. The safety profile of CAT-152 was similar to that of placebo. PMID- 17908592 TI - Interface fluid syndrome in human eye bank corneas after LASIK: causes and pathogenesis. AB - PURPOSE: To evaluate the effects of corneal edema on human donor corneas that had previous LASIK using a laboratory model with histologic and ultrastructural correlations. DESIGN: Experimental study. PARTICIPANTS: Thirty human eye bank corneas from 15 donors (mean age +/- standard deviation, 49.9+/-8.9 years) who had had previous LASIK surgery (2-8 years before death). METHODS: The corneas were mounted in an artificial anterior chamber and the corneal endothelium was perfused for up to 5.0 hours with 0.9% saline solution (endothelial cell damage group) or BSS Plus at a pressure of 15 mmHg (control group), or BSS Plus at a pressure of 55 mmHg (high-pressure group). The corneas were evaluated by confocal and specular microscopy before, during, and at the end of the experimental period. Subsequently, the specimens were evaluated by light and electron microscopy. MAIN OUTCOME MEASURES: Corneal thickness, reflectivity, histology, and ultrastructure. RESULTS: Endothelial cell damage resulted in an increased (141.5+/-38.8 microm) total corneal thickness relative to controls (52.3+/-33.7 microm), whereas high pressure resulted in a decreased thickness (24.8+/-14.1 microm) relative to controls. This ultimately was due to swelling of the LASIK interface in both groups and swelling of the residual stromal bed (RSB) in the endothelial cell damage group or compression of the RSB and, possibly, the flap in the high-pressure group. A significant increase in corneal reflectivity at the LASIK interface occurred in both groups, primarily due to varying degrees of fluid accumulation and associated hydropic keratocyte degeneration, as well as increased corneal reflectivity in the RSB only in the endothelial cell damage group. CONCLUSIONS: After LASIK surgery, edematous corneas preferentially hydrate and swell in the paracentral and central interface wound, commonly resulting in a hazy corneal appearance primarily due to keratocyte hydropic degeneration. More severe corneal edema is characterized by the formation of an optically empty space corresponding to an interface fluid pocket. The spectrum of interface fluid syndrome can be described in 3 stages. PMID- 17908593 TI - Steroidal and nonsteroidal antiinflammatory medications can improve photoreceptor survival after laser retinal photocoagulation. AB - OBJECTIVE: To determine whether methylprednisolone or indomethacin can enhance photoreceptor survival after laser retinal injury in an animal model. DESIGN: Experimental study. PARTICIPANTS: Twenty rhesus monkeys. METHODS: Twenty rhesus monkeys (Macaca mulatta) received a grid of argon green (514.5 nm, 10 ms) laser lesions in the macula of the right eye and a grid of neodymium:yttrium-aluminum garnet (Nd:YAG; 1064 nm, 10 ns) lesions in the macula of the left eye, followed by randomization to 2 weeks of treatment in 1 of 4 treatment groups: high-dose methylprednisolone, moderate-dose methylprednisolone, indomethacin, or control. The lesions were assessed at day 1, day 14, 2 months, and 4 months. The authors were masked to the treatment group. This report discusses the histologic results of ocular tissue harvested at 4 months. MAIN OUTCOME MEASURE: The number of surviving photoreceptor cell nuclei within each lesion was compared with the number of photoreceptor nuclei in surrounding unaffected retina. The proportion of surviving photoreceptor nuclei was compared between each treatment group. RESULTS: Argon retinal lesions in the high-dose steroid treatment group and the indomethacin treatment group demonstrated improved photoreceptor survival compared with the control group (P = 0.004). Hemorrhagic Nd:YAG lesions demonstrated improved survivability with indomethacin treatment compared with controls (P = 0.003). In nonhemorrhagic Nd:YAG laser retinal lesions, the lesions treated with moderate-dose steroids demonstrated improved photoreceptor survival compared with the control group (P = 0.004). CONCLUSIONS: Based on histologic samples of retinal laser lesions 4 months after injury, treatment with indomethacin resulted in improved photoreceptor survival in argon laser lesions and hemorrhagic Nd:YAG laser lesions. Treatment with systemic methylprednisolone demonstrated improved photoreceptor survival in argon retinal lesions and in nonhemorrhagic Nd:YAG lesions. PMID- 17908594 TI - Adverse events after intravitreal triamcinolone in patients with and without uveitis. AB - PURPOSE: To evaluate the rates of adverse ocular events after intravitreal triamcinolone acetonide (IVTA) injection in patients with and without uveitis. DESIGN: Retrospective observational case series. PARTICIPANTS: Two hundred twenty two eyes of 173 patients were included in the study: 45 eyes of 31 patients with macular edema (ME) due to uveitis and 177 eyes of 142 patients with ME secondary to other etiologies. METHODS: Retrospective review of patients who received IVTA at the Cole Eye Institute for ME attributable to various causes between the years 2001 and 2005. Data review of clinical records included patient demographics, etiology of ME, and adverse outcomes after injection. Rates of adverse outcomes in patients with and without uveitis were compared. MAIN OUTCOME MEASURES: Intraocular pressure (IOP) elevation and posterior subcapsular cataract (PSC) progression. RESULTS: Uveitis patients were significantly younger, more likely to be female, and more likely to have had prior posterior sub-Tenon's capsule steroid injection and/or glaucoma therapy than their nonuveitis counterparts. In a multivariate analysis adjusting for the differences in these factors, the presence of uveitis was the strongest risk factor for an adverse IOP event (odds ratio, 2.5; 95% confidence interval [CI], 1.0-6.1; P = 0.05). The odds of having a documented increase in PSC after IVTA injection were 5.6 times greater in uveitis eyes (P = 0.007; 95% CI, 1.6-19.6). CONCLUSIONS: Intraocular pressure elevation and PSC progression occurred with greater frequency in uveitis patients receiving IVTA. Patients with uveitis treated with IVTA should be counseled about these risks and monitored closely. PMID- 17908596 TI - CAT-152 Trabeculectomy Study. PMID- 17908595 TI - Optic nerve head and retinal nerve fiber layer analysis: a report by the American Academy of Ophthalmology. AB - OBJECTIVE: To evaluate the current published literature on the use of optic nerve head (ONH) and retinal nerve fiber layer (RNFL) measurement devices in diagnosing open-angle glaucoma and detecting progression. METHODS: A search of peer-reviewed literature was conducted on February 15, 2006 in PubMed and the Cochrane Library for the period January 2003 to February 2006. The search was limited to studies of adults in English-language journals and yielded 442 citations. The panel reviewed the abstracts of these articles and selected 159 articles of possible clinical relevance for review. Of these 159 full-text articles, 82 were determined to be relevant for the first author and methodologist to review and rate according to the quality of evidence. RESULTS: There were no studies classified as having the highest level of evidence (level I). The ONH and RNFL imaging instruments reviewed in this assessment were determined to be highly effective in distinguishing eyes with glaucomatous visual field (VF) loss from normal eyes without VF loss, based on level II evidence. In addition, some studies demonstrated that parameters from ONH or RNFL imaging predicted the development of VF defects among glaucoma suspects. Studies on detecting glaucoma progression showed that although there was often agreement on progression between the structural and functional (VF) tests, a significant proportion of glaucoma patients progressed by either the structural or the functional test alone. CONCLUSIONS: The ONH and RNFL imaging devices provide quantitative information for the clinician. Based on studies that have compared the various available technologies directly, there is no single imaging device that outperforms the others in distinguishing patients with glaucoma from controls. Ongoing advances in imaging and related software, as well as the impracticalities associated with obtaining and assessing optic nerve stereophotographs, have made imaging increasingly important in many practice settings. The information obtained from imaging devices is useful in clinical practice when analyzed in conjunction with other relevant parameters that define glaucoma diagnosis and progression. PMID- 17908597 TI - Bevacizumab for graft rejection. PMID- 17908598 TI - Anterior stromal puncture after LASIK. PMID- 17908599 TI - Penetrating keratoplasty for keratoconus. PMID- 17908600 TI - Recurrent corneal erosion. PMID- 17908604 TI - Macular OCT changes after vitrectomy. PMID- 17908602 TI - Vascular endothelial growth factor. PMID- 17908605 TI - Classification of macular holes. PMID- 17908606 TI - Eye health in East Timor. PMID- 17908608 TI - Urate deposits. PMID- 17908611 TI - Evisceration vs. enucleation. PMID- 17908612 TI - Evisceration vs. enucleation. PMID- 17908614 TI - Bacterial eradication rates with shortened courses of 2nd- and 3rd-generation cephalosporins versus 10 days of penicillin for treatment of group A streptococcal tonsillopharyngitis in adults. AB - In a meta-analysis of 5 randomized controlled trials involving 1030 adults, the likelihood of bacteriologic eradication in the treatment of group A beta hemolytic streptococcal (GAS) tonsillopharyngitis with 5 days of select cephalosporins (cefpodoxime, cefuroxime, cefotiam, and cefdinir) was noninferior to 10 days of penicillin (odds ratio, 1.46; 95% confidence interval, 0.96-2.22, P = 0.08). PMID- 17908615 TI - Detection of the host immune response to Burkholderia mallei heat-shock proteins GroEL and DnaK in a glanders patient and infected mice. AB - We examined, by enzyme-linked immunosorbent assay and Western blot analysis, the host immune response to 2 heat-shock proteins (hsps) in a patient and mice previously infected with Burkholderia mallei. The patient was the first reported human glanders case in 50 years in the United States. The expression of the groEL and dnaK operons appeared to be dependent upon a sigma(32) RNA polymerase as suggested by conserved heat-shock promoter sequences, and the groESL operon may be negatively regulated by a controlling invert repeat of chaperone expression (CIRCE) site. In the antisera, the GroEL protein was found to be more immunoreactive than the DnaK protein in both a human patient and mice previously infected with B. mallei. Examination of the supernatant of a growing culture of B. mallei showed that more GroEL protein than DnaK protein was released from the cell. This may occur similarly within an infected host causing an elevated host immune response to the B. mallei hsps. PMID- 17908616 TI - Molecular characterization of class 1 integrons and antimicrobial resistance in Aeromonas strains from foodborne outbreak-suspect samples and environmental sources in Taiwan. AB - One hundred thirty-three Aeromonas spp. isolates were examined for multiple antibiotic resistance phenotypes and prevalence of class 1 integron sequences. Twenty-four (18.0%) of these isolates contained class 1 integron. Seven different class 1 integrons were found among 24strains, with a total of 10 different gene cassettes encoding for resistance to trimethoprim (dfr12 and dfr2d), aminoglycosides (aadA1 and aadA2), beta-lactam antibiotics (oxa2), chloramphenicol (catB3 and catB8), quaternary ammonium amines (qacE2), and 2 ORFs (orfD and orfF) with unknown function. Rate of antibiotic resistance was different between integron-positive and integron-negative strains. Trimethoprim and trimethoprim-sulphamethoxazole resistances were commonly associated with integron, and all of integron-positive isolates were multiple resistant to more than 3 agents. Resistance to as many as 10 antimicrobial agents were observed in integron-positive strains. Several cassette arrays of class 1 integrons identified in this study were not previously reported in Aeromonas strains. This study demonstrates the wide distribution of class 1 integron in Aeromonas spp. isolated from foodborne outbreak-suspect samples and environmental sources in Taiwan. PMID- 17908617 TI - Zyvox Annual Appraisal of Potency and Spectrum Program Results for 2006: an activity and spectrum analysis of linezolid using clinical isolates from 16 countries. AB - The Zyvox Annual Appraisal of Potency and Spectrum Program has completed its fifth year of monitoring for emerging resistance to linezolid and other Gram positive active agents on the continents of Europe, Asia, Australia, and Latin America. In 2006, 4216 Gram-positive isolates from 16 nations were submitted for analysis from 6 organism groups including Staphylococcus aureus (54.0%), coagulase-negative staphylococci (CoNS) (14.6%), enterococci (10.0%), Streptococcus pneumoniae (9.4%), viridans group streptococci (5.0%), and beta hemolytic streptococci (7.0%). Linezolid retained potent activity against S. aureus (MIC(50) and MIC(90), 2 microg/mL; 39.8% methicillin resistant) and CoNS (MIC(50) and MIC(90), 1 microg/mL; 74.3% methicillin resistant). Despite endemicity of vancomycin-resistant enterococci (up to 30.0%) in several nations, linezolid inhibited >99% of strains at or= 60 years (OR, 1.68; 95% CI, 1.262.23; p < 0.001), inadequate bowel preparation (OR, 1.66; 95% CI, 1.212.16; p = 0.001), history of constipation (OR, 2.43; 95% CI, 1.045.69; p = 0.042), and body mass index (BMI) < 25 kg/m2 (OR, 1.41; 95% CI, 1.051.89; p = 0.024) were all independent risk factors. The probability of incomplete insertion was significantly associated with the sum of the aforementioned risk factors (p < 0.001). Compared with subjects with no risk factors, the risks of incomplete insertions increased significantly among subjects bearing 1 risk factor (OR, 2.57; 95% CI, 1.474.49; p = 0.001), 2 risk factors (OR, 4.41; 95% CI, 2.527.39; p < 0.001), 3 risk factors (OR, 6.40; 95% CI, 3.5611.52; p < 0.001) and >or= 4 risk factors (OR, 10.00; 95% CI, 3.8925.70; p < 0.001). CONCLUSION: Female sex, age >or= 60 years, BMI < 25 kg/m2, history of constipation, and inadequate bowel preparation were independent risk factors for incomplete insertion of flexible sigmoidoscopy. Subjects with multiple risk factors may consider alternative modalities for colonic examination. PMID- 17908650 TI - Gastrointestinal stromal tumors: computed tomographic features and prediction of malignant risk from computed tomographic imaging. AB - BACKGROUND: Gastrointestinal stromal tumors (GISTs) are specific, generally Kit (CD117)-positive, mesenchymal tumors of the gastrointestinal tract encompassing a majority of tumors previously considered gastrointestinal smooth muscle tumors. Our aim was to characterize the computed tomographic findings and predict malignant risk from computed tomography for the evaluation of GISTs. METHODS: The computed tomographic images of 39 patients with pathologically and immunohistochemically proven GISTs were reviewed by 2 radiologists, and the final interpretations were reached by consensus. Images were assessed for the size, contour, growth pattern, boundary, degree of enhancement, and necrosis of the tumors. The presence of calcification within the lesions, abdominal lymphadenopathy, ascites, and bowel obstruction were also recorded. Categorical variables were compared using Fishers exact test. Univariate and multivariate logistic regression analyses were used for selection of significant predictors of high-risk malignancy. In addition, the relationships between computed tomographic features and tumor size were assessed by means of nonparametric univariate analysis with the MannWhitney U test and KruskalWallis test. RESULTS: Both old age and larger tumor size (>or= 5 cm) were statistically significant in the univariate logistic analysis for high-risk malignant tumors (p < 0.25). However, in multivariate logistic regression, only larger tumor size (>or= 5 cm) was found to have final statistical significance for high-risk malignant GISTs (p < 0.05). In addition, more exophytic growth pattern (p < 0.01), more lobulated appearance (p < 0.01), good enhancement (p < 0.05),and more necrosis (p < 0.01) of masses were more often observed in larger GISTs than small ones on computed tomography. CONCLUSION: Larger tumor size (>or= 5 cm) was found to have a predictive value with respect to high-risk malignant GISTs. PMID- 17908651 TI - The clinical manifestations and risk factors of a delayed diagnosis of Kawasaki disease. AB - BACKGROUND: Kawasaki disease (KD) is an acute febrile vasculitis and may cause coronary artery abnormalities. Due to the higher incidence in Asian countries, most pediatricians in Taiwan are familiar with KD. However, there are still some patients being diagnosed 10 days after the onset of the illness and not receiving a highly effective therapy. In this study, we analyzed the risk factors and clinical manifestations of patients with a delayed diagnosis of KD. METHODS: A retrospective review was made of the medical records of the patients diagnosed with KD at our institution between January 1996 and December 2005. The patients were divided into 2 groups: early-diagnosis group (EDG: diagnosis was made within 10 days after the onset of the fever) and delayed-diagnosis group (DDG: diagnosis was made 10 days after the onset of the fever). RESULTS: Fourteen of a total of 78 children (17.9%) were grouped into the DDG group, and 64 into the EDG group. There were no statistical differences between the 2 groups in terms of age, gender, number of antibiotics used, day of the first medical visit, total days of skin rash, conjunctivitis, mucosa changes, lymphadenopathy or laboratory examinations except for the higher white blood cell count and serum immunoglobulin G level in the DDG group. The patients in the EDG group had a clustered onset of symptoms as compared to the DDG group with a dispersed and late onset of symptoms. There was a higher risk of coronary artery abnormalities in the DDG group than the EDG group (42.9% vs. 14.1%; p = 0.036), and in the patients with KD who were younger than 1 year (29.0% vs. 12.7%; p = 0.043). CONCLUSION: Patients with delayed diagnosis of KD were associated with higher risk of developing coronary arterial lesions. It is necessary to develop a diagnostic test for KD and provide more education to health care providers for early recognition of KD. PMID- 17908652 TI - Expression of vascular endothelial growth factor in Taiwanese benign and malignant prostate tissues. AB - BACKGROUND: The expression of vascular endothelium growth factor (VEGF) has been correlated to the grading and stage of prostate cancers. However, data regarding Taiwanese prostate cancer patients are lacking. The aim of the present study was to examine VEGF expression in our radical prostatectomy specimens. METHODS: Fifty one radical prostatectomy specimens with prostate cancer (15 stage pT2N0, 25 pT3N0, 11 pT2-4 N1) were stained using goat anti-human VEGF polyclonal antibody (AB-293NA; R&D Systems Inc., Minneapolis, MN, USA). The VEGF expression in malignant and nonmalignant prostate tissues was compared. The correlations of VEGF immunoreactivity with Gleason scores and pathologic stages were examined. MannWhitney U test was used for comparison of preoperative prostate-specific antigen levels between patients with and without VEGF expression. RESULTS: Positive VEGF staining was observed in 80.4% of malignant epithelia, 39.2% of peritumoral stroma, 68.6% of benign hyperplastic glands, and 25.5% of adjacent stroma. There was no difference in VEGF expression between malignant and nonmalignant areas. Advanced disease had significantly higher frequency of stroma but not epithelium VEGF staining as compared to organ-confined disease (p = 0.002 and p = 0.412, respectively). The Gleason 7 and higher tumors had significantly higher frequency of VEGF staining in stroma but not glandular epithelium (p = 0.041 and p = 0.353, respectively). Tumors with positive epithelium VEGF staining had significantly higher PSA levels (21.3 18.1 vs. 10.8 6.8 ng/mL; p = 0.013). CONCLUSION: There was no difference in VEGF immunoreactivity between malignant and benign prostatic epithelium in Taiwanese. High Gleason grade tumors and advanced disease had significantly higher frequency of VEGF expression in stroma but not glandular epithelium. Tumors with positive epithelium VEGF staining had significantly higher PSA levels. PMID- 17908653 TI - Is ward experience in resuscitation effort related to the prognosis of unexpected cardiac arrest? AB - BACKGROUND: The aim of this study was to investigate the outcome of patients of unexpected cardiac arrest initially resuscitated by first responders with dissimilar experiences under the support of cardiac arrest team (CAT). METHODS: All unexpected cardiac arrest patients receiving in-hospital resuscitation with the activation of CAT in a tertiary-care teaching hospital over a 12-month period were recorded according to the Utstein criteria. We prospectively recorded various factors at resuscitation and retrospectively evaluated the outcome. Outcome measures included return of spontaneous circulation (ROSC), survival longer than 24 hours, and survival to discharge. RESULTS: Altogether, 76 emergency calls were registered, and among these, 44 calls (58%) were cardiac arrests, including 8 ventricular tachycardia/fibrillation, 15 pulseless electrical activity, and 21 asystole. The rate of ROSC was 61%, the rate of survival longer than 24 hours was 37%, and the rate of survival to discharge was 18%. The response time of our CAT was 271 seconds (4 minutes and 31 seconds) on average. The patients who collapsed in the wards experienced in resuscitation effort received higher rates of appropriate basic and advanced cardiac life support interventions before CAT arrival (79% vs. 44%; p = 0.019), had an increased chance of ROSC (75% vs. 38%; p = 0.014), survival longer than 24 hours (54% vs. 13%; p = 0.007), and survival to discharge (29% vs. 0%; p = 0.036). CONCLUSION: Hospital wards with more than 5 cardiac arrests per year have a better patient survival rate than those with fewer arrests. This is despite all ward staff receiving the same level of training. PMID- 17908654 TI - Pituitary apoplexy after thyrotropin-releasing hormone stimulation test in a patient with pituitary macroadenoma. AB - Pituitary apoplexy is a rare complication of pituitary tumors. We report a case of a 41-year-old female with acromegaly due to a pituitary macroadenoma, who developed pituitary apoplexy after a thyrotropin-releasing hormone (TRH) 200 microgram intravenous injection stimulation test. Neither emergency computed tomography (CT) scans nor magnetic resonance imaging (MRI), performed 6 hours and 12 hours, respectively, after the active episode, disclosed the evidence of acute hemorrhage or infarction. Two days later, the pituitary mass, removed by transsphenoidal approach, showed ischemic necrosis and acute hemorrhage. The TRH test is generally safe for evaluating pituitary function, but pituitary apoplexy may occur after the procedure. CT and MRI may miss the diagnosis of pituitary apoplexy, especially if performed immediately after the acute episode. PMID- 17908656 TI - Endoscopic intubation with aid of mechanical ventilation via a dedicated nasopharyngeal airway. AB - A young child with jaw-neck-sternum immobility suffering from acute upper airway obstruction was treated with nasotracheal intubation using flexible endoscope (FE). During this difficult intubation, an inserted trimming endotracheal tube acted as a nasopharyngeal airway and simultaneous supplement with mechanical ventilation through the tube successfully resuscitated and improved the patients ventilation and oxygenation. This management can greatly facilitate visualization of the laryngeal apparatus and translaryngeal passage of the FE. This technique can be helpful in resuscitative ventilation and difficult intubation in a critical upper airway emergency. PMID- 17908655 TI - Rapidly progressive glomerulonephritis associated with nontuberculous mycobacteria. AB - A 72-year-old woman with a past medical history of nontuberculous mycobacteria (NTM) pulmonary disease was admitted because of hemoptysis and acute renal failure. A chest X-ray showed interstitial infiltration over bilateral lung fields. Kidney biopsy showed immune complex-mediated acute diffuse proliferative glomerulonephritis with 48% crescents and glomerular endocapillary hypercellularity with exudative neutrophils suggestive of infection-related glomerulonephritis. Reactivated NTM infection of the lungs was suspected when mycobacterial cultures of the sputum repeatedly yielded Mycobacterium avium. A lung biopsy revealed chronic inflammation without evidence of alveolar capillaritis. A diagnosis of NTM pulmonary disease was further confirmed by tissue culture of the lung biopsy specimens. Antituberculous drugs in combination with clarithromycin were given for the treatment of NTM infection. Pulmonary symptoms promptly responded to the treatments. Furthermore, renal function steadily improved after initiation of anti-NTM therapy. To our knowledge, this is the first report of rapidly progressive glomerulonephritis associated with NTM infection. PMID- 17908657 TI - Bilateral renal cell carcinoma in a patient with autosomal dominant polycystic kidney disease. AB - Renal cell carcinoma (RCC) in autosomal dominant polycystic kidney disease (ADPKD) is very rare. Only 11 cases of bilateral RCC in ADPKD have been reported since 1954. Herein, we present a 58-year-old male who received laparoscopic bilateral radical nephrectomy for bilateral RCC with different cell variants in ADPKD and end-stage renal disease under regular hemodialysis. PMID- 17908658 TI - The current role of 1.5T non-contrast 3D time-of-flight magnetic resonance angiography to detect intracranial steno-occlusive disease. AB - BACKGROUND/PURPOSE: This study was performed to evaluate the role of non-contrast 3D time-of-flight (TOF) magnetic resonance angiography (MRA) to detect and quantify intracranial steno-occlusive disease. METHODS: Between April 2004 and January 2006, 45 patients with both 1.5T TOF MRA and digital subtraction angiography (DSA) performed within a 30-day interval were included. We evaluated the following intracranial arterial segments: petrous internal carotid artery (ICA), cavernous ICA, supraclinoid ICA, M1 of middle cerebral artery, A1 of anterior cerebral artery, P1 of posterior cerebral artery, basilar artery, and distal vertebral artery. In total, 675 arterial segments were evaluated and categorized as negative, moderate-1 (3049% stenosis), moderate-2 (5069%), severe (7099% stenosis, including gap sign on MRA), and occlusion. RESULTS: The sensitivity and specificity of TOF MRA for > 29% stenosis and > 49% stenosis were 94%, 96% and 95%, 96%, respectively; while sensitivity and specificity for occlusion lesions were both 100%. However, 44 segments (37% of diseased segments) were overestimated by MRA, including 20 false-positive stenoses (which occurred in 10 [22%] patients) and 24 overestimated stenosis degree. The gap sign as severe stenosis only showed about 21% sensitivity and 41% specificity. Seven lesions were underestimated by MRA: three arterial segments were out of the field of MRA examination, and four were moderate-1stenosis on DSA. CONCLUSION: TOF MRA has high sensitivity and specificity in detecting all categories of stenosis degree and occlusion. However, it tends to overestimate lesions. Therefore, MRA can be considered as a screening study. Confirmation with other studies is recommended in doubtful cases. PMID- 17908659 TI - Sonographic imaging of meniscal subluxation in patients with radiographic knee osteoarthritis. AB - BACKGROUND/PURPOSE: This study was undertaken to describe the sonographic features of meniscal subluxation in the weight-bearing position and to determine any association between meniscal subluxation and radiographic osteoarthritis. METHODS: In total, 238 knees with symptoms were examined successfully with weight bearing anteroposterior and lateral radiographs and high resolution ultrasonography. The radiographs were examined to determine whether participants had radiographic osteoarthritis, graded using the Kellgren-Lawrence Scale. The degree of subluxation of the medial meniscus in each knee was measured using high resolution ultrasound with a 10-MHz linear transducer, at the level of the medial collateral ligament in weight-bearing condition. The degree of subluxation was compared in knees with the presence or absence of radiographic osteoarthritis using Students t test. Additional analysis between knees with early and advanced radiographic osteoarthritis was also performed. RESULTS: Meniscal subluxation for knees with (n = 141) and without (n = 97) radiographic signs of osteoarthritis were 4.3 1.9 mm and 0.7 0.6 mm, respectively. The difference was highly significant (p < 0.001). After age adjustment, the medial meniscal subluxation of age-matched subjects were 4.8 1.7 mm for knees with radiographic osteoarthritis (n = 43) and 1.0 0.8 mm for knees without such changes (n = 43). The difference between the two groups was still significant (p < 0.001). The greatest meniscal subluxation was seen in knees with advanced radiographic signs of osteoarthritis; no knee with osteoarthritic changes on radiographs had an undisplaced meniscus. CONCLUSION: Meniscal subluxation is a prominent feature on weight-bearing sonographic imaging in patients with radiographic osteoarthritis and could be considered as a risk factor for the development of knee osteoarthritis. By using musculoskeletal ultrasonography, one can detect this occult meniscal derangement early before the appearance of radiographic signs of osteoarthritis. PMID- 17908660 TI - Effects of prone position on inflammatory markers in patients with ARDS due to community-acquired pneumonia. AB - BACKGROUND/PURPOSE: Acute respiratory distress syndrome (ARDS) is a serious disorder of intensive care unit patients. We evaluated the safety of continuous prone position ventilation (PRONE) and its effects on oxygenation and plasma cytokine concentrations in patients with ARDS caused by severe community-acquired pneumonia (CAP). METHODS: This was a prospective observational clinical study conducted in a respiratory intensive care unit of a 1200-bed medical center in central Taiwan. Twenty-two patients with severe CAP and ARDS were included. They were treated by traditional supine ventilation (SUPINE, n = 11) or PRONE (n = 11) if they met the criteria for ARDS. Patients in the PRONE group were ventilated in prone position continuously for at least 72 hours. Plasma cytokines were collected and analyzed at baseline, 24 hours and 72 hours after enrolment. Serial PaO2/FiO2 and complications were evaluated. RESULTS: Complications associated with PRONE were minor and self-limited. PRONE had higher PaO2/FiO2 ratio than SUPINE did at 48 hours after enrolment. The levels of plasma IL-6 concentration declined significantly with time in the PRONE group (p = 0.011). The levels of plasma IL-6 concentration at enrolment, 24 hours and 72 hours after enrolment also predicted the 14th day mortality of all patients. CONCLUSION: PRONE was a safe and effective maneuver for improving oxygenation in patients with severe CAP and ARDS. PRONE also influenced IL-6 expression in patients with severe CAP. PMID- 17908661 TI - Comparison between pancreaticojejunostomy and pancreaticogastrostomy after pancreaticoduodenectomy. AB - BACKGROUND/PURPOSE: Pancreatic leakage is a leading cause of morbidity and mortality after pancreaticoduodenectomy (PD). Pancreaticogastrostomy (PG) has been reported to be associated with a lower pancreatic leakage rate and morbidity rate than pancreaticojejunostomy (PJ). This study compared the preoperative characteristics, surgical risk factors, intraoperative parameters, and postoperative outcome between PJ and PG. METHODS: From March 1992 to March 2005, a comparative study between PJ and PG for patients with periampullary lesions undergoing PD was conducted. A total of 377 consecutive patients underwent PD. Among them, 188 patients underwent PJ and 189 underwent PG. RESULTS: The overall mortality, morbidity and pancreatic leakage following PD were 5%, 45.1% and 10.6%, respectively. The mortality, morbidity and pancreatic leakage were 8.9%, 56.4% and 17.6% in the PJ group, and 2.1%, 33.9% and 3.7% in the PG group (p < 0.001). Mean operative time was 9.3 hours versus 6.7 hours (p < 0.001), mean blood loss was 1032 mL versus 891 mL (p = 0.064) and mean hospital stay was 34.8 days versus 26.1 days (p < 0.001) in the PJ and PG groups, respectively. PJ, soft pancreas, pancreatic duct stenting, low surgical volume (< 20) and age (> 65 years) were identified as risk factors for pancreatic leakage, while PJ, soft pancreas, pancreatic duct stenting and low surgical volume (< 20) were four significant risk factors for surgical morbidity. Further, PJ, pancreatic leakage, low surgical volume (< 20) and age (> 65 years) were identified to be surgical risk factors for mortality. CONCLUSION: PG is a safer method than PJ following PD as a significantly lower rate of pancreatic leakage, surgical morbidity and mortality, shorter operation time, and shorter postoperative hospital stay are reported. PMID- 17908662 TI - Fine needle aspiration cytology stained with Rius method in quicker diagnosis of medullary thyroid carcinoma. AB - BACKGROUND/PURPOSE: The cytologic features of medullary thyroid carcinoma (MTC) have been described and documented in the literature. Most of the studies were related to the Papanicolaou stain or the May-Grnwald-Giemsa stain. The aim of the present study was to analyze detailed cytologic characteristics of MTC diagnosed by fine-needle aspiration cytology (FNAC) stained with a quick method, Rius stain. METHODS: We collected the FNAC slides of 38 cases with MTC that were proven by surgical pathology. The cytologic findings were reviewed under a light microscope. The median age at diagnosis was 46.5 years, ranging from 13 to 83 years; 20 were women and 18 were men. Among them, seven cases were familial forms and the others were sporadic. RESULTS: The majority of these cases showed moderate to abundant cellularity. Small round cells, spindle-shaped cells and large oval to polygonal cells formed the usual components. Twenty-seven cases were categorized as pleomorphic cell type and 11 cases as monomorphic type. Except in one case, cohesive and clustered small round cells predominated in all cases, alone or coupled with cohesive spindle cells and/or scattered large oval to polygonal cells. The diagnosis of mixed medullary-follicular thyroid carcinoma was made in the case composed solely of grouped polygonal cells. Cytoplasmic granularities were noted in 14 cases and vacuolations were seen in eight. Binucleated and multinucleated cells were not uncommon, while intranuclear cytoplasmic inclusions were less frequent. The diagnostic sensitivity for MTC by FNAC was 89%. CONCLUSION: The distinctive cytopathologic characteristics of MTC by Rius stain allowed us to make the diagnosis in aspiration biopsies accurately and quickly. PMID- 17908663 TI - Efficacy of mammographic evaluation of breast cancer in women less than 40 years of age: experience from a single medical center in Taiwan. AB - BACKGROUND/PURPOSE: Mammography is the standard imaging modality for breast cancer diagnosis. However, the value of mammographic diagnosis in breast cancer patients aged less than 40 years old has not been well assessed. The goal of our study was to determine the diagnostic efficacy of mammography for the detection of breast cancer in women under 40 years of age in a single medical center in Taiwan. METHODS: Of 1766 women diagnosed with breast cancer in one medical center between 1999 and 2005, 227 (12.9%) who were younger than 40 years of age were enrolled, and 105 of these 227 patients had pre-biopsy mammograms available for analysis. The sensitivities for mammography at first (prospective) and second (retrospective) readings and for corresponding ultrasound were calculated. The distribution of different breast composition between the mammographic true positive (TP) and false-negative (FN) lesions at the first and second readings was analyzed. RESULTS: Of the 105 patients, 104 presented with a palpable mass and the other one was asymptomatic. There were 109 pathologically proven breast cancers from the 105 patients; 92 of 109 cancerous lesions were detected at the first mammographic reading (sensitivity 84.4%), and the most common mammographic sign was microcalcifications (40.2%). The second reading detected seven additional cancers (99 of 109 lesions; sensitivity 90.8%). There was no significant difference between mammographic TP and FN lesions for the different breast composition on first and second readings. Ninety patients also had ultrasound available for correlation with 94 cancers diagnosed from them. The diagnostic sensitivity of ultrasound was 94.7% (89 of 94 lesions). CONCLUSION: Mammography has an acceptable sensitivity for the detection of breast cancer in women aged less than 40 years, regardless of different breast composition. Breast ultrasound can offer a higher sensitivity for such a population. PMID- 17908664 TI - Incidence and related factors of violence in emergency departments--a study of nurses in southern Taiwan. AB - BACKGROUND/PURPOSE: Nurses in emergency departments are high-risk groups who are most likely to experience the offense of violent behaviors. In a comparison on the occurrence of verbal and physical abuses in emergency departments, this study aimed to analyze the correlation of staff properties and occupational conditions in an attempt to provide a reference on violence prevention. METHODS: Eleven hospitals in southern Taiwan were selected and a survey of 267 nurses was conducted. The results were analyzed by percentage, Pearsons chi2 test and logistic regression. RESULTS: The 236 valid questionnaires showed that nurses experienced significantly more verbal abuse (92%) than physical abuse (30%). The highest occurrence of verbal abuse was 49% in night shift, and the physical abuse was 44% in overnight shift. Long waiting (89%), difference in cognition (87%), and lack of communication (82%) were the common causes. The occurrences of verbal abuse and physical abuse were significantly correlated to less-informed (odds ratio [OR], 1.67 and 1.22, respectively) and basic-level nurses (OR, 2.30 and 1.34, respectively). In addition, younger age (OR, 2.80; p < 0.01) and single status (OR, 9.09; p <0.05) were correlated with occurrence of verbal abuse, but not of physical abuse. The test on occupational conditions showed a significant correlation (OR, 0.68/0.44; p < 0.005) between the occurrence of verbal/physical abuse and whether supervisors could provide enough training to cope with violence. CONCLUSION: This study suggested that a well-informed nurse should be arranged to take the night/overnight shifts in order to reduce the occurrence of abuses in emergency departments. PMID- 17908665 TI - Prediction for major adverse outcomes in cardiac surgery: comparison of three prediction models. AB - BACKGROUND/PURPOSE: Recent advances in medical treatment have altered the profile of patients referred for cardiac surgery. The proportion of high risk patients has increased dramatically. Numerous multifactorial risk scores have been developed to predict outcomes after cardiac surgery. However, these additive risk models were all developed outside of Asia and have never been validated in Taiwan. We applied the Parsonnet score, Tu score and logistic regression to a population in Taiwan who received cardiac surgery to predict the mortality, morbidity and likelihood of prolonged stay in the intensive care unit (ICU). METHODS: This retrospective study included 622 adult patients who received cardiac surgery during a 2-year period at Taichung Veterans General Hospital. The patients were randomly divided into a reference set (n = 423) and a validation set (n = 199). The Parsonnet score and Tu score were calibrated separately with the reference set to determine mortality, morbidity and likelihood of prolonged ICU stay. We developed a separate logistic regression model for each of the three outcomes by using the reference set. The validation set was used to test these models. RESULTS: The area under the receiver operating characteristic (ROC) curve (AUC) of the Parsonnet score, Tu score and logistic regression for predicting in hospital mortality were 0.843, 0.714 and 0.867, respectively. The AUC of the Parsonnet score, Tu score and logistic regression for predicting major morbidity were 0.784, 0.736 and 0.808, respectively. The AUC of the Parsonnet score, Tu score and logistic regression for predicting likelihood of prolonged ICU stay were 0.701, 0.689 and 0.764, respectively. CONCLUSION: The Parsonnet score performed as well as the logistic regression models in predicting major adverse outcomes. The Parsonnet score appears to be a very suitable model for clinicians to use in risk stratification of cardiac surgery. PMID- 17908666 TI - Paraneoplastic Pemphigus and Bronchiolitis Obliterans in a Patient with Splenic B cell Lymphoma. AB - Paraneoplastic pemphigus (PNP), also called paraneoplastic autoimmune multiorgan syndrome, is a rare disorder associated with underlying neoplasia. The common underlying neoplasms include non-Hodgkins lymphoma, chronic lymphocytic leukemia, and Castlemans disease. Though B-cell lymphoma is the most common underlying malignancy, only one case associated with splenic B-cell lymphoma has been recognized. The prognosis of PNP is very poor, and PNP-associated bronchiolitis obliterans (BO) is not uncommon. Herein, we report a 44-year-old woman who initially presented with multiple oral ulcers, conjunctivitis, and numerous cutaneous blisters. Serial workup established the diagnosis of PNP and revealed an underlying splenic B-cell lymphoma. Although the mucocutaneous lesions gradually healed after splenectomy and chemotherapy, deteriorating respiratory function developed 7 months later with pathologically proven BO. She finally succumbed to respiratory failure 12 months after presentation despite intensive respiratory care. PMID- 17908667 TI - Hyperhomocysteinemia, deep vein thrombosis and vitamin B12 deficiency in a metformin-treated diabetic patient. AB - Vitamin B12 deficiency may be induced by long-term use of metformin, which may in turn lead to hyperhomocysteinemia. Thus, hyperhomocysteinemia may increase the risk of vascular thrombosis in diabetic patients, when metformin is used and a homozygous methylenetetrahydrofolate reductase (MTHFR) C677T mutation is present. We report a 65-year-old Taiwanese diabetic woman who was treated with metformin for 6 years and who had suffered from swelling of the left lower extremity for 3 months. Ascending venography confirmed the diagnosis of proximal deep vein thrombosis, while hyperhomocysteinemia, megaloblastic anemia caused by vitamin B12 deficiency, and a homozygous C677T mutation of the MTHFR gene were also found. She had no identifiable venous thrombotic risk factors other than hyperhomocysteinemia, which seemed to be caused by both MTHFR C677T homozygous mutation and vitamin B12 deficiency. With the substitution of insulin injection for metformin, short-term supplement of vitamin B12, and anticoagulant therapy for the deep vein thrombosis, her anemia and hyperhomocysteinemia recovered rapidly. The deep vein thrombosis also responded well. Our findings highly suggested the role of metformin in causing vitamin B12 deficiency, which may serve as an additional risk factor for venous thrombosis in diabetic patients. Our report also highlights the need to check vitamin B12 levels during metformin treatment. PMID- 17908668 TI - Acquisition and cure of autoimmune disease following allogeneic hematopoietic stem cell transplantation. AB - Hematopoietic stem cell transplantation (HSCT) can either cause or eliminate autoimmune disease. Here, we report two cases. One was a 33-year-old woman with myelodysplastic syndrome (refractory anemia) who received bone marrow transplantation from her human leukocyte antigen (HLA)-identical sister who had a history of Graves disease. Antithyroid antibodies, including antimicrosomal antibody and antithyroglobulin antibody, appeared 4 months after transplantation. Clinical hyperthyroidism appeared 7 months after transplantation, and a hypothyroid state was noted 2 months later. The other case was a 50-year-old woman with Sjgrens syndrome and hypothyroidism who was diagnosed with peripheral T cell non-Hodgkins lymphoma. She received allogeneic peripheral blood stem cell transplantation (PBSCT) from her histocompatible sister owing to only partial response to traditional chemotherapy. Cure of lymphoma and remission of Sjgrens syndrome was noted 4 years after PBSCT. These two illustrative cases, one of acquisition of hyperthyroidism and the other of remission of Sjgrens syndrome after transplantation, highlights that HSCT can induce adoptive autoimmune disease or cure coincidental autoimmune disease. Donor selection and attentive monitoring is required in such circumstances. PMID- 17908669 TI - Urologic manifestations of acute appendicitis secondary to metastatic cervical cancer. AB - Acute abdominal pain may be attributed to a variety of medical or surgical conditions. Acute appendicitis, a common entity in differential diagnosis, may present with diverse clinical manifestations. It may occasionally mimic urogenital disorders and be particularly challenging to diagnose in women. We report a 34-year-old woman who had undergone radical hysterectomy 2 years previously for stage Ib cervical cancer. She presented with lower abdominal pain, dysuria, and fever of 2 days duration, unrelieved by 5 days of antibiotics. Computed tomography revealed an enlarged appendix surrounded by an abscess, and appendectomy was performed. Pathologic examination of the surgical specimen revealed metastatic cervical cancer in the appendix. Patients with acute appendicitis may manifest with urologic disorders that can be caused by metastatic tumor. PMID- 17908670 TI - Diabetic ketoacidosis in a patient with acromegaly. AB - Diabetes mellitus develops in about 10% of acromegalic patients, usually secondary to insulin resistance caused by growth hormone excess. Diabetic ketoacidosis is a result of relative insulin deficiency and is a rare feature of acromegaly. Here, we present one case of this disorder. A 57-year-old man came to the emergency room due to 2 weeks of dizziness. He also had polyuria, polydipsia, nausea, diplopia, blurred vision and dysarthria. His plasma glucose level was 32.06 mmol/L, plasma osmolarity was 322 mOsm/L, arterial pH was 7.30, level of bicarbonates was 18 mmol/L, urine ketones was 4+, and HbA1c was 14.1%. No specific cause for the development of this metabolic derangement could be found. He displayed clinical features of acromegaly during admission, which was confirmed by an elevated growth hormone level and pituitary macroadenoma shown on magnetic resonance imaging. The patient underwent total transsphenoid tumor removal 2 weeks later; plasma glucose levels became normal thereafter. PMID- 17908671 TI - Application of forensic entomology to postmortem interval determination of a burned human corpse: a homicide case report from southern Taiwan. AB - Determining the postmortem interval (PMI) is strongly impacted by several variables, which consequently results in inaccuracy in the estimation of PMI used in court trials. A PMI experiment was conducted in Kaohsiung County by disposing a burned pig corpse in the woods. One month later, unexpectedly and interestingly, a homicide case, very similar to this mock study, occurred at a distance of 6 km away from the experimental site. The female victim had been killed and burned. The maggots collected from the victim were identified to be Chrysomya megacephala by morphologic observation and were then confirmed by mitochondrial DNA sequence. A PMI of 50 hours was concluded for the burned human body, based on the information of the maggots from the pig corpse. The murderer was eventually arrested and confessed to the crime. According to his statement, the elapsed time since death was calculated to have been 46 hours. In this case, the PMI was estimated successfully and it was almost precise. It would appear that the more similar the surrounding environment between the mock study and the actual case, the more precise can be the PMI estimation. PMID- 17908672 TI - Performance as annihilation or integration? AB - What happens when the analyst has the impression of being annihilated by the patient? Analysts have a tendency to use more general, i.e. simplifying, constructions such as destructiveness, psychosis or death instinct as explanatory models. In the authors' view, these constructions in the end evade rather than mirror clinical reality. More recent research points to promising possibilities of differentiation, e.g. psychotic mechanisms which are--as yet undiscussed- based on Freud's notion of the partial 'rent in the relation between ego and external world'. These findings emphasize the restitutive function of a symptom or disturbance, i.e. destruction of a relationship which hinders the therapeutic process and which is not understood initially, instead of solely stressing the destructive meaning in a tabooing gesture. The concept of performance attempts to replace simplifying models with a discriminant process, and will be preliminarily defined and explained in delineation to terms already in use such as acting out, enactment, and role responsiveness. The authors explore the question of how the perception of unthought certainty in the performance can either be recognized as a blueprint, i.e. organizing activity, or as the destruction of the relationship so that a new one can emerge. The evidence from a detailed clinical example shows that many treatments can fail at this point and demonstrates how an understanding of performance in this sense offers a chance for integrating processes that otherwise impede treatment. PMID- 17908673 TI - The intersubjective links in perversion. AB - The author studies the intersubjective links which the pervert maintains with analyst or partner, attempting to indicate the differences between the investments in each case. Rather than accepting that empathy towards these patients is impossible to achieve and disturbs the countertransference profoundly, it attempts to show that these difficulties may be overcome if they are reinterpreted in the light of the theory of the intersubjective link. The author examines the theories and the practice of intersubjectivity and gives a definition of his approach to the link between two subjects. He applies these ideas to the case of a sexually masochistic female patient. The countertransference is marked successively by indifference, rejection and smothering. The analysis of the analyst's dream allows the situation to evolve. Failures in primary identification can result in domination over others and utilitarianism. The author examines the place of the challenge to the 'Law' and the father (in the attempt by the patient to put a theory to the test) in order to identify the figure of the witness in the pervert's intersubjective links. The desire of the transference would be marked by the figure of the witness rather than by that of the analyst as accomplice. PMID- 17908674 TI - Bodily centered protections in adolescence: an extension of the work of Frances Tustin. AB - In this paper, the author discusses the recurrence of infantile, proto-mental functioning in adolescence mainly in the context of the work of Frances Tustin. She demonstrates, through clinical example, how the tendency to resort to bodily centered and sensation-dominated protections is reactivated on a grand scale when the internal and external physical and psychological changes, brought on in puberty, are felt to be potentially overwhelming. She also demonstrates how, when the capacity for adequate mental and emotional development is stultified, sensation and action once again come to the rescue as the adolescent's way of attenuating anxieties unconsciously experienced as resonating with those unmentalized happenings of early infancy and how the psychoanalytic relationship may be pivotal in setting previously derailed mental and emotional growth back on track. PMID- 17908675 TI - On the edge: the psychoanalyst's transference. AB - Countertransference is a central topic in analytic work and in the literature. The concept of countertransference includes a basic question which has been understood in different ways. The author attempts to differentiate between the psychoanalyst's transference and his countertransference in the analytic process. It is hard to draw a line between them; analysts are always on the edge. The analyst's transference will be explored and described using three approaches: narcissism, regression profile and the analyst's phase of life. Regression profile is a new concept developed by the author, which may help us to understand the core of the analyst's transference in the analytic situation. She illustrates the topic by clinical vignettes. PMID- 17908676 TI - Elements of analytic style: Bion's clinical seminars. AB - The author finds that the idea of analytic style better describes significant aspects of the way he practices psychoanalysis than does the notion of analytic technique. The latter is comprised to a large extent of principles of practice developed by previous generations of analysts. By contrast, the concept of analytic style, though it presupposes the analyst's thorough knowledge of analytic theory and technique, emphasizes (1) the analyst's use of his unique personality as reflected in his individual ways of thinking, listening, and speaking, his own particular use of metaphor, humor, irony, and so on; (2) the analyst's drawing on his personal experience, for example, as an analyst, an analysand, a parent, a child, a spouse, a teacher, and a student; (3) the analyst's capacity to think in a way that draws on, but is independent of, the ideas of his colleagues, his teachers, his analyst, and his analytic ancestors; and (4) the responsibility of the analyst to invent psychoanalysis freshly for each patient. Close readings of three of Bion's 'Clinical seminars' are presented in order to articulate some of the elements of Bion's analytic style. Bion's style is not presented as a model for others to emulate or, worse yet, imitate; rather, it is described in an effort to help the reader consider from a different vantage point (provided by the concept of analytic style) the way in which he, the reader, practices psychoanalysis. PMID- 17908677 TI - Semiotic transformations in psychoanalysis with infants and adults. AB - The author addresses issues that emerge when we compare psychoanalytic experiences with adults and with infants. Two analyses-one with a 35 year-old woman and one with a 2 week-old boy and his mother-illustrate that infant psychoanalytic experiences help us understand and handle adult transference. However, we cannot extrapolate infant experiences to adult work. Truly, witnessing the baby's communication widens our sensitivity to non-verbal layers of the adult's communication. Infant work also offers a direct encounter with the container and the contained personified by a mother with her baby. But we need to conceptualize carefully the links between clinical experiences with babies and adults. When we call an adult transference pattern 'infantile', we imply that primeval experience has been transformed into present behaviour. However, if we view the analytical situation as one in which infantile invariants have transformed into adult symptoms, we face the impossible task of indicating the roots of the present symptoms. The author rather suggests that what is transformed is not an invariant infantile essence but signs denoting the patient's inner reality. He proposes we define transformation as a semiotic process instead of building it on an essentialist grounding. If we view the analytic situation as a map of signs that we translate during our psychoanalytic work, we can proceed into defining containment as a semiotic process. This idea will be linked with a conceptualization of the mother-infant relation in semiotic terms. PMID- 17908678 TI - A perfectly staged 'concerted action' against psychoanalysis: the 1913 congress of German psychiatrists. AB - An eyewitness account provides evidence of a significant clandestine effort to neutralize the legitimacy and authority of psychoanalysis. In a letter, the witness confirms the existence of a perfectly staged concerted action among German psychiatrists against Freud's influence in 1913. Their congress in Breslau was meant to present the united front of German psychiatrists, who were going on record as being against psychoanalysis and, in that context, to give Eugen Bleuler, a leading psychiatrist, whose (however half-hearted) support for psychoanalysis had alarmed his colleagues, a public opportunity for back pedalling. The letter shows that Freud and his allies were not the only ones who tried to manage an intellectual movement by using informal networks and 'behind the scenes' manoeuvring. PMID- 17908679 TI - Authors who have an impact on candidates' training: cultural differences and theoretical languages. AB - The authors consider the influence that a sense of geographical and cultural ties of candidates from different regions has on their theoretical interests. They question the way that this is taken into consideration in psychoanalytic training. The function of theory, both in terms of its transmission and the creation of new knowledge, is explored from this perspective. The results of an Internet survey are presented. The candidate sample for this survey (N = 250) was drawn from Europe, Latin America and North America, and candidates were asked to indicate their degree of interest for each of the 55 authors in a given list. The results showed that there were significant differences in the areas of theoretical interest of the candidates depending on the geographical region. Furthermore, what is also significant is how these differences in areas of theoretical interest were linked to those authors who had developed their work in the same geographical region as the candidates. These differences are shown to be connected to the candidates' sense of regional belonging. Data are also presented about which authors have the greatest impact in a given region, along with the influence values of the authors in relation to each one of the regions. Finally, the candidates' interest in each of the authors is specified in terms of a general mean rank and a regional mean rank, thus showing which authors candidates find most interesting in each of the regions. The study concludes by arguing that the results of the investigation enable us to question how psychoanalytic theory is transmitted, and, more specifically, how it is transmitted within institutions at a regional level. It is also suggested that the means be found to uncover the inconsistencies linked to cultural ties. It is proposed that further research be conducted to look more deeply into how cultural differences play a part in the different theoretical languages in the training of psychoanalysts. PMID- 17908680 TI - Supervision, transference and countertransference. AB - The author discusses supervision, transference and countertransference as seen in the context of the clinical case of a patient who had been first seen as a training analysis case and who later, in a fortuitous way, was treated by the supervisor of the training analysis. The supervisor, who in the first instance did not recognize the patient, discusses the reasons for this unusual experience in terms of the presence and absence of transference during the analysis of this patient as a training case and the problems inherent in the task of supervising. The patient's feelings towards the first and the second analyst and the vicissitudes of transference and countertransference during the supervision of the training analysis and its influence on the presentation of the analytical sessions by the student are also detailed and discussed. The question of recorded supervision presentations and their possible influence on the dynamics of supervision is raised. PMID- 17908681 TI - Losing a training analyst for ethical violations: a candidate's perspective. AB - The author presents her experience as the analysand of a training analyst who was investigated and expelled for ethical violations with another patient, including sexual-boundary violations, during her analytic training. While boundary violations by training analysts are not uncommon, the particular trauma experienced by 'bystanders' such as candidates and supervisees is not discussed in the literature, nor the response of institutes to the educational problems that are generated. The author illustrates the complications for candidates that arise from the dual roles of training analyst as educator and analyst when he or she faces investigation or censure, including isolation and secrecy, which promote various splits in the candidate, analytic dyad and group, as well as loyalty conflicts. The discussion covers three phases of the author's experience as a candidate-analysand, namely the period encompassing the institute's ethics investigation, the announcement of findings to her and to the institute as a group, and the ensuing individual and group dynamics generated by her analyst's expulsion from the institute and revocation of his medical license. Theoretical perspectives are utilized to understand the group regression, including contamination and contagion fears, which occurred in the wake of the training analyst's expulsion, and the impact of these processes on the candidate, including the pressure to function as a 'container' for projections of the group. Implications and recommendations for candidates and institutes are made for dealing helpfully with trainees who are affected by the process of dealing with a training analyst's ethical violations. Short-term and longer-term outcomes of the experience are considered. PMID- 17908682 TI - On: projective identification. PMID- 17908684 TI - Observations on the calcium dependence and reversibility of cobalamin transport across the outer membrane of Escherichia coli. AB - The calcium dependence of cobalamin (Cbl) binding to the BtuB protein of Escherichia coli and the reversibility of its function in the transport of Cbl across the outer membrane have been examined. The results show that the two calcium-binding sites in BtuB that were identified previously by others are responsible for the calcium dependence of high affinity Cbl binding. The affinity of the pure BtuB protein for Cbl was approximately 1000-fold higher in the presence of saturating levels of calcium than in its absence. The affinities of BtuB for both Cbl and calcium were decreased by insertion of alanine residues at position 51 of the mature protein and were increased by several mutations and deletions in the TonB box. Experiments on the uptake of Cbl into the periplasmic space showed that this process is reversible and that the exit of Cbl back into the medium does not require the protonmotive force. Our interpretation of these results is that the role of the TonB-ExbB-ExbD complex, potentiated by the protonmotive force, is to reduce the affinity of the Cbl-binding site, thus increasing the rate of Cbl release into the periplasmic space. The evidence also indicates that access of the Cbl-binding site of BtuB to the periplasmic space does not require removal of the hatch domain from the barrel. PMID- 17908685 TI - The role of the N terminus and transmembrane domain of TRPM8 in channel localization and tetramerization. AB - Transient receptor potential (TRP) channels are a family of cation channels involved in diverse cellular functions. They are composed of a transmembrane domain of six putative transmembrane segments flanked by large N- and C-terminal cytoplasmic domains. The melastatin subfamily (TRPM) channels have N-terminal domains of approximately 700 amino acids with four regions of shared homology and C-terminal domains containing the conserved TRP domain followed by a coiled-coil region. Here we investigated the effects of N- and C-terminal deletions on the cold and menthol receptor, TRPM8, expressed heterologously in Sf21 insect cells. Patch-clamp electrophysiology was used to study channel activity and revealed that only deletion of the first 39 amino acids was tolerated by the channel. Further N-terminal truncation or any C-terminal deletions prevented proper TRPM8 function. Confocal microscopy with immunofluorescence revealed that amino acids 40-86 are required for localization to the plasma membrane. Furthermore, analysis of deletion mutant oligomerization shows that the transmembrane domain is sufficient for TPRM8 assembly into tetramers. TRPM8 channels with C-terminal deletions tetramerize and localize properly but are inactive, indicating that although not essential for tetramerization and localization, the C terminus is critical for proper function of the channel sensor and/or gate. PMID- 17908686 TI - The CheC phosphatase regulates chemotactic adaptation through CheD. AB - The bacterial chemotaxis system is one of the most extensively studied signal transduction systems in biology. The response regulator CheY controls flagellar rotation and is phosphorylated by the CheA histidine kinase to its active form. CheC is a CheY-P phosphatase, and this activity is enhanced in a CheC-CheD heterodimer. CheC is also critical for chemotactic adaptation, the return to the prestimulus system state despite persistent attractant concentrations. Here, CheC point mutants were examined in Bacillus subtilis for in vivo complementation and in vitro activity. The mutants were identified separating the three known abilities of CheC: CheD binding, CheY-P binding, and CheY-P phosphatase activity. Remarkably, the phosphatase ability was not as critical to the in vivo function of CheC as the ability to bind both CheY-P and CheD. Additionally, it was confirmed that CheY-P increases the affinity of CheC for CheD, the later of which is known to be necessary for receptor activation of CheA. These data suggest a model of CheC as a CheY-P-induced regulator of CheD. Here, CheY-P would cause CheC to sequester CheD from the chemoreceptors, inducing adaptation of the chemotaxis system. This model represents the first plausible means for feedback from the output of the system, CheY-P, to the receptors. PMID- 17908687 TI - Endotoxin priming of neutrophils requires NADPH oxidase-generated oxidants and is regulated by the anion transporter ClC-3. AB - Several soluble mediators, including endotoxin, prime neutrophils for an enhanced respiratory burst in response to subsequent stimulation. Priming of neutrophils occurs in vitro, and primed neutrophils are found in vivo. We previously localized the anion transporter ClC-3 to polymorphonuclear leukocytes (PMN) secretory vesicles and demonstrated that it is required for normal NADPH oxidase activation in response to both particulate and soluble stimuli. We now explore the contribution of the NADPH oxidase and ClC-3 to endotoxin-mediated priming. Lipooligosaccharide (LOS) from Neisseria meningitidis enhances the respiratory burst in response to formyl-Met-Leu-Phe, an effect that was impaired in PMNs lacking functional ClC-3 and under anaerobic conditions. Mobilization of receptors to the cell surface and phosphorylation of p38 MAPK by LOS were both impaired in PMN with the NADPH oxidase chemically inhibited or genetically absent and in cells lacking functional ClC-3. Furthermore, inhibition of the NADPH oxidase or ClC-3 in otherwise unstimulated cells elicited a phenotype similar to that seen after endotoxin priming, suggesting that basal oxidant production helps to maintain cellular quiescence. In summary, NADPH oxidase activation was required for LOS-mediated priming, but basal oxidants kept unstimulated cells from becoming primed. ClC-3 contributes to both of these processes. PMID- 17908688 TI - Neuronal glutamate transporters vary in substrate transport rate but not in unitary anion channel conductance. AB - Excitatory amino acid transporters (EAATs) not only sustain a secondary active glutamate transport but also function as anion-selective ion channels. The relative proportion of currents generated by glutamate transport or by the chloride conductance varies for each cloned EAAT subtype. For EAAT1, EAAT2, and EAAT3, the anion current is only a small component of the total transporter associated current amplitude, whereas EAAT4 and EAAT5 transporters mediate predominantly anion currents. We here demonstrate that the distinct current proportions are entirely due to differences in glutamate transport rates. EAAT3 and EAAT4 differ in unitary glutamate transport rates as well as in the voltage and substrate dependence of anion channel opening, but ion conduction properties are very similar. Noise analysis revealed identical unitary current amplitudes and similar absolute open probabilities for the two anion channels. The low glutamate transport rate of EAAT4 allows regulation of cellular excitability without interfering with extracellular glutamate homeostasis and makes this EAAT isoform ideally suited to regulate excitability in dendritic spines of Purkinje neurons. PMID- 17908689 TI - Kruppel-like factor 4 is acetylated by p300 and regulates gene transcription via modulation of histone acetylation. AB - Colon cancer is the second leading cause of cancer death in the United States. Kruppel-like factor 4 (KLF4) is a transcription factor involved in both proliferation and differentiation in the colon. It is down-regulated in both mouse and human colonic adenomas and has been implicated as a tumor suppressor in the gut, whereas in breast cancer, KLF4 is an oncogene. KLF4 is also involved in reprogramming differentiated cells into pluripotent stem cells. KLF4 can act as a transcriptional activator or repressor, but the underlying mechanisms are poorly understood. We found that p300, a CREB-binding protein-related protein, interacts with KLF4 both in vitro and in vivo and activates transcription. We further made the novel observation that endogenous KLF4 is acetylated by p300/CBP in vivo and that mutations of the acetylated lysines resulted in a decreased ability of KLF4 to activate target genes, suggesting that acetylation is important for KLF4 mediated transactivation. Furthermore, we found that KLF4 differentially modulates histone H4 acetylation at the promoters of target genes. Co transfection of KLF4 and HDAC3 resulted in a synergistic repression of a cyclin B(1) reporter construct. Our results suggest that KLF4 might function as an activator or repressor of transcription depending on whether it interacts with co activators such as p300 and CREB-binding protein or co-repressors such as HDAC3. PMID- 17908692 TI - The advantages of being locked. Assessing the cleavage of short and long RNAs by locked nucleic acid-containing 8-17 deoxyribozymes. AB - RNA-cleaving deoxyribozymes can be used for the sequence-specific knockdown of mRNAs. It was previously shown that activity of these deoxyribozymes is enhanced when their substrate-binding arms include some locked nucleic acid (LNA) residues, but the mechanistic basis of this enhancement was not explored. Here we dissected the kinetics and thermodynamics underlying the reaction of LNA containing 8-17 deoxyribozymes. Four 8-17 constructs were designed to target sequences within the E6 mRNA from human papillomavirus type 16. When one of these deoxyribozymes (DNAzymes) and the corresponding LNA-armed enzyme (LNAzyme) were tested against a minimal RNA substrate, they showed similar rates of substrate binding and similar rates of intramolecular cleavage, but the LNAzyme released its substrate more slowly. The superior thermodynamic stability of the LNAzyme substrate complex led to improved performances in reactions carried out at low catalyst concentrations. The four DNAzymes and the corresponding LNAzymes were then tested against extended E6 transcripts (>500 nucleotides long). With these structured substrates, the LNAzymes retained full activity, whereas the DNAzymes cleaved extremely poorly, unless they were allowed to pre-anneal to their targets. These results imply that LNAzymes can easily overcome the kinetic barrier represented by local RNA structure and bind to folded targets with a faster association rate as compared with DNAzymes. Such faster annealing to structured targets can be explained by a model whereby LNA monomers favor the initial hybridization to short stretches of unpaired residues ("nucleation"), which precedes disruption of the local mRNA structure and completion of the binding process. PMID- 17908690 TI - Controlling the inhibition of the sarcoplasmic Ca2+-ATPase by tuning phospholamban structural dynamics. AB - Cardiac contraction and relaxation are regulated by conformational transitions of protein complexes that are responsible for calcium trafficking through cell membranes. Central to the muscle relaxation phase is a dynamic membrane protein complex formed by Ca2+-ATPase (SERCA) and phospholamban (PLN), which in humans is responsible for approximately 70% of the calcium re-uptake in the sarcoplasmic reticulum. Dysfunction in this regulatory mechanism causes severe pathophysiologies. In this report, we used a combination of nuclear magnetic resonance, electron paramagnetic resonance, and coupled enzyme assays to investigate how single mutations at position 21 of PLN affects its structural dynamics and, in turn, its interaction with SERCA. We found that it is possible to control the activity of SERCA by tuning PLN structural dynamics. Both increased rigidity and mobility of the PLN backbone cause a reduction of SERCA inhibition, affecting calcium transport. Although the more rigid, loss-of function (LOF) mutants have lower binding affinities for SERCA, the more dynamic LOF mutants have binding affinities similar to that of PLN. Here, we demonstrate that it is possible to harness this knowledge to design new LOF mutants with activity similar to S16E (a mutant already used in gene therapy) for possible application in recombinant gene therapy. As proof of concept, we show a new mutant of PLN, P21G, with improved LOF characteristics in vitro. PMID- 17908691 TI - Rapamycin promotes vascular smooth muscle cell differentiation through insulin receptor substrate-1/phosphatidylinositol 3-kinase/Akt2 feedback signaling. AB - The phenotypic plasticity of mature vascular smooth muscle cells (VSMCs) facilitates angiogenesis and wound healing, but VSCM dedifferentiation also contributes to vascular pathologies such as intimal hyperplasia. Insulin/insulin like growth factor I (IGF-I) is unique among growth factors in promoting VSMC differentiation via preferential activation of phosphatidylinositol 3-kinase (PI3K) and Akt. We have previously reported that rapamycin promotes VSMC differentiation by inhibiting the mammalian target of rapamycin (mTOR) target S6K1. Here, we show that rapamycin activates Akt and induces contractile protein expression in human VSMC in an insulin-like growth factor I-dependent manner, by relieving S6K1-dependent negative regulation of insulin receptor substrate-1 (IRS 1). In skeletal muscle and adipocytes, rapamycin relieves mTOR/S6K1-dependent inhibitory phosphorylation of IRS-1, thus preventing IRS-1 degradation and enhancing PI3K activation. We report that this mechanism is functional in VSMCs and crucial for rapamycin-induced differentiation. Rapamycin inhibits S6K1 dependent IRS-1 serine phosphorylation, increases IRS-1 protein levels, and promotes association of tyrosine-phosphorylated IRS-1 with PI3K. A rapamycin resistant S6K1 mutant prevents rapamycin-induced Akt activation and VSMC differentiation. Notably, we find that rapamycin selectively activates only the Akt2 isoform and that Akt2, but not Akt1, is sufficient to induce contractile protein expression. Akt2 is required for rapamycin-induced VSMC differentiation, whereas Akt1 appears to oppose contractile protein expression. The anti restenotic effect of rapamycin in patients may be attributable to this unique pattern of PI3K effector regulation wherein anti-differentiation signals from S6K1 are inhibited, but pro-differentiation Akt2 activity is promoted through an IRS-1 feedback signaling mechanism. PMID- 17908693 TI - Dimerization of Hsp90 is required for in vivo function. Design and analysis of monomers and dimers. AB - Heat shock protein 90 (Hsp90) plays a central role in signal transduction and has emerged as a promising target for anti-cancer therapeutics, but its molecular mechanism is poorly understood. At physiological concentration, Hsp90 predominantly forms dimers, but the function of full-length monomers in cells is not clear. Hsp90 contains three domains: the N-terminal and middle domains contribute directly to ATP binding and hydrolysis and the C domain mediates dimerization. To study the function of Hsp90 monomers, we used a single-chain strategy that duplicated the C-terminal dimerization domain. This novel monomerization strategy had the dual effect of stabilizing the C domain to denaturation and hindering intermolecular association of the ATPase domain. The resulting construct was predominantly monomeric at physiological concentration and did not function to support yeast viability as the sole Hsp90. The monomeric construct was also defective at ATP hydrolysis and the activation of a kinase and steroid receptor substrate in yeast cells. The ability to support yeast growth was rescued by the addition of a coiled-coil dimerization domain, indicating that the parental single-chain construct is functionally defective because it is monomeric. PMID- 17908694 TI - NADPH oxidase activity selectively modulates vascular endothelial growth factor signaling pathways. AB - Vascular endothelial growth factor (VEGF) and reactive oxygen species (ROS) play critical roles in vascular physiology and pathophysiology. We have demonstrated previously that NADPH oxidase-derived ROS are required for VEGF-mediated migration and proliferation of endothelial cells. The goal of this study was to determine the extent to which VEGF signaling is coupled to NADPH oxidase activity. Human umbilical vein endothelial cells and/or human coronary artery endothelial cells were transfected with short interfering RNA against the p47(phox) subunit of NADPH oxidase, treated in the absence or presence of VEGF, and assayed for signaling, gene expression, and function. We show that NADPH oxidase activity is required for VEGF activation of phosphoinositide 3-kinase-Akt forkhead, and p38 MAPK, but not ERK1/2 or JNK. The permissive role of NADPH oxidase on phosphoinositide 3-kinase-Akt-forkhead signaling is mediated at post VEGF receptor levels and involves the nonreceptor tyrosine kinase Src. DNA microarrays revealed the existence of two distinct classes of VEGF-responsive genes, one that is ROS-dependent and another that is independent of ROS levels. VEGF-induced, thrombomodulin-dependent activation of protein C was dependent on NADPH oxidase activity, whereas VEGF-induced decay-accelerating factor-mediated protection of endothelial cells against complement-mediated lysis was not. Taken together, these findings suggest that NADPH oxidase-derived ROS selectively modulate some but not all the effects of VEGF on endothelial cell phenotypes. PMID- 17908695 TI - Autologous HSCT for advanced MS: is the glass half-empty or really half-full? PMID- 17908696 TI - Responses to desiccation stress in bryophytes and an important role of dithiothreitol-insensitive non-photochemical quenching against photoinhibition in dehydrated states. AB - The effects of air drying and hypertonic treatments in the dark on seven bryophytes, which had grown under different water environments, were studied. All the desiccation-tolerant species tested lost most of their PSII photochemical activity when photosynthetic electron transport was inhibited by air drying, while, in all the sensitive species, the PSII photochemical activity remained at a high level even when photosynthesis was totally inhibited. The PSI reaction center remained active under drying conditions in both sensitive and tolerant species, but the activity became non-detectable in the light only in tolerant species due to deactivation of the cyclic electron flow around PSI and of the back reaction in PSI. Light-induced non-photochemical quenching (NPQ) was found to be induced not only by the xanthophyll cycle but also by a DeltapH-induced, dithiothreitol-insensitive mechanism in both the desiccation-tolerant and intolerant bryophytes. Both mechanisms are thought to have an important role in protecting desiccation-tolerant species from photoinhibition under drying conditions. Fluorescence emission spectra at 77K showed that dehydration-induced quenching of PSII fluorescence was observed only in tolerant species and was due to neither state 1-state 2 transition nor detachment of light-harvesting chlorophyll protein complexes from PSII core complexes. The presence of dehydration-induced quenching of PSI fluorescence was also suggested. PMID- 17908697 TI - Seroprevalence of HIV and other vertically transmitted viral infections among pregnant women of Amritsar (Punjab, North India). PMID- 17908698 TI - Prevalence of celiac disease, Helicobacter pylori and gastroesophageal reflux in patients with refractory iron deficiency anemia. AB - OBJECTIVE: The aim of this article is to determine the prevalence of celiac disease (CD), Helicobacter pylori (H. pylori) and gastroesophageal reflux (GER) in patients with resistant iron deficiency anemia (IDA). PATIENTS: The study included 25 patients <18 years of age with refractory IDA (not responding to iron therapy for 3 months in a dose of 6 mg elemental iron/kg/day). METHODS: All patients included in the study were subjected to careful history taking and thorough clinical examination. Blood sample was taken for analysis of antibodies for CD including: antigliadin antibody (AGA), antiendomysial antibody (EMA), antireticulin antibody (ARA) and antitissue Transglutaminase (tTg) IgG antibody. Anti-H. pylori IgG antibodies and a (13)C-urea breath test (UBT) was done to all patients to diagnose H. pylori. Upper gastrointestinal tract endoscopy was done for all patients to evaluate for the presence of some etiologies of intractable anemia as chronic blood loss. These included: CD, H. pylori infection and GER. The upper gastrointestinal tract endoscopy was also done to evaluate the presence of bleeding spots, ulcers or angiomatous malformations. In addition, gastric antral biopsies were taken for diagnosis of H. pylori infection by the following tests: rapid urease test, histopathological examination and culture. RESULTS: CD was positive in 11 out of 25 patients (44%), H. pylori infection in 12 out of 25 patients (48%), while GER was diagnosed in 11 out of 25 patients (44%). Patients with CD had age of presentation < or =2 years in two patients (18.2%) while the remaining nine patients (81.8%) had age of presentation >2 years and it was statistically significant (p = 0.05*). Also patients with H. pylori had age of presentation < or =4 years in five patients (41.7%) and the remaining seven patients (81.8%) had age of presentation >4 years and it was statistically significant (p = 0.03*). Logistic regression analysis demonstrated that the risk factors for severity of anemia were age of patients and duration of anemia. On the other hand, other parameters have no significant influence on the severity of anemia. Also risk factors of short stature were age of presentation of anemia, degree of anemia and H. pylori infection. AGA had the highest sensitivity (100%) followed by antiendomysium antibody (81.8%) while the tTG antibody had the highest specificity (85.7%) for diagnosis of CD. UBT and histopathology had the highest sensitivity (100%) for diagnosis of H. pylori while rapid urease test, culture, H. pylori stool antigen and anti-H. pylori IgG antibody had the highest specificity (100%). In conclusion, refractory IDA may be due to clinically unapparent H. pylori gastritis and CD. CD is one of the most common causes of intestinal malabsorption during childhood which leads to impairment of iron absorption. Apart from offering them gluten-free diet rich in iron, early detection and treatment of IDA and prophylactic iron and folic acid supplementation will go a long way to optimize their mental and psychological functions. Eradication of H. pylori infection with concomitant iron therapy should correct the anemia. PMID- 17908699 TI - Use evidence to expose the unequal distribution of problems and the unequal distribution of solutions. PMID- 17908700 TI - A case of rheumatoid vasculitis involving the gastrointestinal tract in early disease. PMID- 17908701 TI - Influenza vaccination in subjects with alpha1-antitrypsin deficiency. AB - BACKGROUND: Influenza vaccination is recommended for all subjects with COPD, including alpha(1)-antitrypsin deficiency (AATD), but immunization practices are below US national goals. Influenza vaccination practices and their relation to respiratory outcomes in AATD are unknown. METHODS: Nine hundred thirty-nine subjects with AATD were followed up prospectively by monthly telephone interviews during the 2003 to 2004 influenza season. Vaccination status, exacerbation rates, and health-care utilization were documented. Residence zip codes were used to group subjects as living in high or low influenza-like illness (ILI) prevalence areas according to published Centers for Disease Control and Prevention data for the same influenza season. RESULTS: Overall, 81.6% of subjects received influenza vaccination, with no differences noted by gender, age (median age 52 years), Global Initiative for Chronic Obstructive Lung Disease stage, or ILI prevalence area. No significant differences were noted in the overall acute exacerbation rates using two different criteria between vaccinated and unvaccinated subjects (mean, 1.5 +/- 1 exacerbations per subject). Similarly, no differences were noted in either the severity of exacerbations or the monthly exacerbation rates between the two groups. Unvaccinated subjects had more unscheduled physician visits than vaccinated subjects, but there were no significant differences in scheduled visits, emergency department visits, or hospitalizations between the two groups. Older age (> 60 years) or residence in a high ILI prevalence area had no effect on outcomes. CONCLUSION: Subjects with AATD in the United States receive adequate influenza vaccination regardless of age. However, we did not observe a significant impact of the vaccination on disease exacerbations and other respiratory outcomes during the 2003 to 2004 influenza season. PMID- 17908702 TI - Photodynamic therapy for airway stenosis in rabbit models. AB - BACKGROUND: Acquired airway stenosis in childhood is resistant to conventional treatment. We examined whether endoscope-assisted photodynamic therapy (PDT) is effective for airway stenosis in animal models of which the pathophysiologic progressions are similar to those of clinical cases showing rapid deterioration. METHODS: Tracheal mucosa-scraped rabbits were administered IV porfimer sodium (Photofrin; Wyeth K.K., Tokyo, Japan) [2 mg/kg], and the tracheal lesions were irradiated with 630 nm of light emitted from a cylindrical diffuser tip via a transtracheal approach. RESULTS: Rabbits without PDT (untreated animals) showed dense granulation tissue in the scraped lesion, resulting in airway stenosis complicated with respiratory stridor. PDT ameliorated the degree of airway stenosis (p = 0.008) and reduced respiratory stridor; rabbits that received PDT showed patchy granulation tissue that was only 20 to 30% of the volume of that seen in the untreated animals. Survival time of rabbits that received PDT was significantly prolonged compared with that of untreated animals (p = 0.03). CONCLUSIONS: PDT was effective for airway stenosis in rabbit models. This suggests that PDT has the potential as a new therapeutic method for airway stenosis originating from granulation tissue. PMID- 17908703 TI - Angiotensin-converting enzyme insertion/deletion polymorphism and risk and outcome of pneumonia. AB - BACKGROUND: Recent studies have suggested involvement of the angiotensin converting enzyme (ACE) insertion/deletion (I/D) polymorphism in the susceptibility to and severity of community-acquired pneumonia (CAP) in Asian populations. We have explored the hypothesis that the ACE I/D polymorphism affects the risk and outcome of CAP in a Dutch white population. METHODS: This is a hospital-based prospective observational study including patients with CAP admitted between October 2004 and August 2006. All patients were genotyped, and pneumonia severity and clinical outcome were compared between patients with II, ID, and DD genotypes of the ACE gene. Pneumonia severity was assessed on day of hospital admission and consecutively on days 2, 3, 5, and 10 of hospital stay using the acute physiology score (APS). Outcomes evaluated were duration of hospital stay, ICU admittance, and in-hospital and 28-day mortality rates. To study the association between ACE genotype and risk of pneumonia, the distribution of the ACE I/D polymorphism was compared with healthy control subjects from the same geographic region. RESULTS: In total, 200 patients with pneumonia and 200 control subjects were included in the study. Mean age of the patients was 63 years. APS scores were not different between the genotype groups on any of the days, and all clinical outcomes (duration of hospital stay, ICU admittance, in-hospital and 28-day mortality rates) were comparable between the three genotype groups. The ACE I/D genotype distribution was identical for patients and control subjects (p = 0.973). CONCLUSIONS: The ACE I/D polymorphism is not associated with risk and outcome of CAP in the Dutch white population. PMID- 17908704 TI - Exposure of airway epithelium to bile acids associated with gastroesophageal reflux symptoms: a relation to transforming growth factor-beta1 production and fibroblast proliferation. AB - RATIONALE: Gastroesophageal reflux (GER) is common in patients with various airway diseases. Airway epithelial cells can release growth factors that promote fibroblast proliferation. Exposure of airway epithelium to bile acids may induce a fibrotic response. OBJECTIVES: To determine how bile acids interact with airway epithelium; particularly, whether transforming growth factor-beta1 secretion and fibroblast proliferation are affected. METHODS: Induced sputum from patients with asthma, GER, or asthma associated with GER symptoms, or from healthy control subjects was collected. Total bile acids were measured by a spectrophotometric enzymatic assay. The major components of bile acids, chenodeoxycholic acid (CD) and glycochenodeoxycholic acid (GCD), were used to stimulate primary airway epithelial cells. Quantitative polymerase chain reaction and Western blotting were applied for messenger RNA expression and signal pathway analysis, respectively. Conditioned medium following CD stimulation was coincubated with fibroblasts for proliferation study. RESULTS: The amount of total bile acids in induced sputum was significantly higher in patients with GER and asthma associated GER symptoms compared to that of healthy control subjects (p<0.005). CD, but not GCD, significantly induced TGF-beta1 production. TGF-beta1 messenger RNA expression was 2.5-fold increased compared to unstimulated cells. This occurred via p38 mitogen-activated protein (MAP) kinase and activating transcription factor-2 activation. Pretreatment with dexamethasone inhibited TGF beta1 production at both messenger RNA and protein levels by inhibiting p38 MAP kinase phosphorylation. Conditioned medium from CD-treated epithelial cells enhanced fibroblast proliferation. CONCLUSIONS: Aspiration of bile acids may induce airway fibrosis through the production of TGF-beta1 and fibroblast proliferation. Early intervention to attenuate these processes may reduce fibrogenesis in various airway diseases associated with GER. PMID- 17908706 TI - Decreased right and left ventricular myocardial performance in obstructive sleep apnea. AB - BACKGROUND: Obstructive sleep apnea (OSA) may predispose patients to congestive heart failure (CHF), suggesting a deleterious effect of OSA on myocardial contractility. METHODS: A cross-sectional study of 85 subjects with suspected OSA who had undergone their first overnight polysomnogram, accompanied by an echocardiographic study. Patients were divided according to the apnea-hypopnea index as follows: < 5 (control subjects); 5 to 14 (mild OSA); and >or= 15 (moderate-to-severe OSA). Right and left ventricular function was evaluated using the myocardial performance index (MPI) and other echocardiographic parameters. For the right ventricle analyses, we excluded patients with a Doppler pulmonary systolic pressure of >or= 45 mm Hg, while for the left ventricle we excluded patients with an ejection fraction of 98th centile) was slightly increased (RR = 1.35, 95% CI = 1.02 to 1.78). CONCLUSIONS: Adoption of the WHO 2006 Growth Charts would set a markedly lower standard of weight gain beyond the age of 4 months for UK infants and could support efforts to avoid future childhood obesity. However, the WHO standard is not representative of size at birth in the UK. PMID- 17908713 TI - Should gonadotropin releasing hormone analogue be administered to prevent premature ovarian failure in young women with systemic lupus erythematosus on cyclophosphamide therapy? PMID- 17908714 TI - Developing evidence-based guidelines for referral for short stature. AB - OBJECTIVE: To establish evidence-based guidelines for growth monitoring on a population basis. STUDY DESIGN: Several auxological referral criteria were formulated and applied to longitudinal growth data from four different patient groups, as well as three samples from the general population. RESULTS: Almost 30% of pathology can be detected by height standard deviation score (HSDS) below -3 or at least two observations of HSDS below -2.5 at a low false-positive rate (<1%) in 0-3-year-old infants. For 3-10-year olds, a rule concerning distance to target height of >2 SD in combination with HSDS <-2.0 has the best predictive value. In combination with a rule on severe short stature (<-2.5 SDS) and a minor contribution from a rule on "height deflection", 85.7% of children with Turner syndrome and 76.5% of children who are short because of various disorders are detected at a false-positive rate of 1.5-2%. CONCLUSIONS: The proposed guidelines for growth monitoring show high sensitivity at an acceptably low false-positive rate in 3-10-year-old children. Distance to target height is the most important criterion. Below the age of 3 years, the sensitivity is considerably lower. The resulting algorithm appears to be suitable for industrialised countries, but requires further testing in other populations. PMID- 17908715 TI - Is there a place for bariatric surgery in treating childhood obesity? PMID- 17908716 TI - Diagnosis dialog: progress report. PMID- 17908717 TI - Invited commentary. PMID- 17908718 TI - On "Development of a clinical prediction rule..." Currier et al. Phys Ther. 2007;87:1106 1119. PMID- 17908719 TI - Potential misdiagnosis of 3-methylcrotonyl-coenzyme A carboxylase deficiency associated with absent or trace urinary 3-methylcrotonylglycine. AB - We report 2 patients with isolated 3-methylcrotonyl-coenzyme A carboxylase deficiency whose urine was devoid of, or contained only trace, 3 methylcrotonylglycine, the pathognomonic marker for this disorder. The first patient, a girl with trisomy 21, was detected through newborn screening with an elevated 5 carbon hydroxycarnitine species level, and the second patient came to clinical attention at the age of 5 months because of failure to thrive and developmental delay. Investigation of urinary organic acids revealed an elevated 3-hydroxyisovaleric acid level but no demonstrable 3-methylcrotonylglycine in both patients. Enzyme studies in cultured fibroblasts confirmed isolated 3 methylcrotonyl-coenzyme A carboxylase deficiency with residual activities of 5% to 7% and 12% of the median control value, respectively. Incorporation of 14C isovaleric acid into intact fibroblasts was essentially normal, showing that the overall pathway was at least partially functional and potentially explaining the absence of 3-methylcrotonylglycine in urine. Mutation analysis of the MCCA and MCCB genes revealed that both patients were compound heterozygous for a missense mutation, MCCB-c.1015G-->A (p.V339M), and a second mutation that leads to undetectable MCCB messenger (poly A+) RNA. Absent or trace 3 methylcrotonylglycine levels in urine raises the potential for misdiagnosis in the clinical biochemical genetics laboratory based solely on urine organic acid analysis using combined gas chromatography-mass spectrometry. PMID- 17908720 TI - Hematopoietic stem cell transplantation corrects the immunologic abnormalities associated with immunodeficiency-centromeric instability-facial dysmorphism syndrome. AB - Immunodeficiency-centromeric instability-facial dysmorphism syndrome, characterized by variable immunodeficiency, centromeric instability, and facial anomalies caused by epigenetic dysregulation resulting in hypomethylation, is caused in many patients by mutations in DNMT3B, a DNA methyltransferase gene; associated infections are a major cause of serious sequelae and death. Hematopoietic stem cell transplantation may improve the clinical course in immunodeficiency-centromeric instability-facial dysmorphism syndrome. We report 3 unrelated patients with persistent infections and intestinal complications who successfully underwent hematopoietic stem cell transplantation after nonmyeloablative or myeloablative conditioning regimens using HLA-matched donors. In all cases, donor chimerism led to resolution of intestinal complications and infections, growth improvement, and correction of the immunodeficiency. PMID- 17908722 TI - Disruptions in insurance coverage: patterns and relationship to health care access, unmet need, and utilization before enrollment in the State Children's Health Insurance Program. AB - BACKGROUND: The numbers and types of disruptions in insurance that children experience and the effects of these disruptions on health care measures have not been well characterized. OBJECTIVES: Our goals were to (1) describe the number and patterns of insurance disruptions within a population of children newly enrolling into the State Children's Health Insurance Program and (2) assess the relationship among insurance disruptions and sociodemographic characteristics of these children and their families to specific measures of access to care, unmet need, and health care utilization during the year before enrollment. METHODS: We conducted telephone interviews in families with children newly enrolling in the State Children's Health Insurance Program. Families reported on measures for each of the 12 months preceding enrollment. They were grouped by number of insurance disruptions in the year before enrollment: continuously uninsured, > or = 2 disruptions, 1 disruption, or continuously insured. RESULTS: Of 920 families contacted, 739 (80%) completed the interview and 710 had useable data. Thirty five percent reported being continuously uninsured, 42% were intermittently insured (> or = 2 disruptions: 28%; 1 disruption: 14%), and 23% were continuously insured during the previous year. The most common patterns of change were between privately insured and uninsured (49%) and Medicaid and uninsured (40%). The continuously uninsured were more likely to be Hispanic and older in age. Multivariate modeling confirmed a gradient between greater insurance disruption and less access to care, less utilization, and greater unmet medical need. Using the continuously uninsured as a reference group, the adjusted odds ratio for having a medical home varied from 2.5 for those with > or = 2 disruptions to 4.5 for the continuously insured and from 1.9 to 3.2, respectively, for using any regular/routine care. The odds ratio for unmet need for a prescription medication was 0.9 for > or = 2 disruptions and 0.5 for those with continuous insurance coverage. CONCLUSIONS: There was significant disruption in insurance coverage in the year before State Children's Health Insurance Program enrollment. Most of these disruptions took the form of children previously enrolled in either Medicaid or private insurance becoming uninsured. Increasing numbers of disruptions were associated with less routine care and greater unmet medical need. These findings suggest that disruptions in insurance coverage for children should be minimized with the adoption of policies regarding continuous eligibility criteria for Medicaid and streamlining transitions between Medicaid, the State Children's Health Insurance Program, and private insurance. PMID- 17908721 TI - Cerebrospinal fluid distribution of ibuprofen after intravenous administration in children. AB - BACKGROUND: Ibuprofen is the most commonly used nonsteroidal, antipyretic, antiinflammatory analgesic in children. Nonsteroidal, antipyretic, antiinflammatory analgesics act in both the peripheral tissues and the central nervous system. The central nervous system penetration of ibuprofen has been described in adults but not in children. OBJECTIVES: Our goals were to investigate the cerebrospinal fluid penetration of ibuprofen in children and evaluate the analgesic plasma concentration of ibuprofen after inguinal surgery in children. MATERIALS AND METHODS: A total 36 healthy children (25 boys) aged 3 months to 12 years received a single intravenous injection of ibuprofen (10 mg/kg). A paired cerebrospinal fluid and blood sample was obtained 10 minutes to 8 hours after the injection. In children having inguinal surgery, a second blood sample was obtained at the time that the child first had wound pain. RESULTS: The ibuprofen level was determined in all cerebrospinal fluid and plasma samples. Cerebrospinal fluid concentrations ranged between 15 and 541 microg/L, and the highest concentrations were measured 30 to 38 minutes after dosing. In all cerebrospinal fluid samples collected after 30 minutes, ibuprofen concentration exceeded that of unbound plasma. The plasma analgesic concentrations after inguinal surgery ranged between 10 and 25 mg/L. CONCLUSIONS: Ibuprofen penetrates the cerebrospinal fluid readily, with peak concentrations attained 30 to 40 minutes after intravenous injection of a 10 mg/kg dose. The plasma analgesic concentration after inguinal surgery with spinal anesthesia is 10 to 25 mg/L. PMID- 17908723 TI - Influence of gender and age on upper-airway length during development. AB - OBJECTIVE: Obstructive sleep apnea has a strong male predominance in adults but not in children. The collapsible portion of the upper airway is longer in adult men than in women (a property that may increase vulnerability to collapse during sleep). We sought to test the hypothesis that in prepubertal children, pharyngeal airway length is equal between genders, but after puberty boys have a longer upper airway than girls, thus potentially contributing to this change in apnea propensity. METHODS: Sixty-nine healthy boys and girls who had undergone computed tomography scans of their neck for other reasons were selected from the computed tomography archives of Rambam and Carmel hospitals. The airway length was measured in the midsagittal plane and defined as the length between the lower part of the posterior hard palate and the upper limit of the hyoid bone. Airway length and normalized airway length/body height were compared between the genders in prepubertal (4- to 10-year-old) and postpubertal (14- to 19-year-old) children. RESULTS: In prepubertal children, airway length was similar between boys and girls (43.2 +/- 5.9 vs 46.8 +/- 7.7 mm, respectively). When normalized to body height, airway length/body height was significantly shorter in prepubertal boys than in girls (0.35 +/- 0.03 vs 0.38 +/- 0.04 mm/cm). In contrast, postpubertal boys had longer upper airways (66.5 +/- 9.2 vs 52.2 +/- 7.0 mm) and normalized airway length/body height (0.38 +/- 0.05 vs 0.33 +/- 0.05 mm/cm) than girls. CONCLUSIONS: Although boys have equal or shorter airway length compared with girls among prepubertal children, after puberty, airway length and airway length normalized for body height are significantly greater in boys than in girls. These data suggest that important anatomic changes at puberty occur in a gender-specific manner, which may be important in explaining the male predisposition to pharyngeal collapse in adults. PMID- 17908724 TI - A geographical comparison of prevalence of overweight school-aged children: the National Survey of Children's Health 2003. AB - OBJECTIVES: This study presents a geographical comparison of state-specific prevalence estimates of children who are at risk of overweight and/or overweight using the 2003 National Survey of Children's Health. METHODS: Using the 2003 National Survey of Children's Health, we computed prevalence estimates of children who are at risk of overweight and/or overweight among a nationally representative sample of 69,000 children between 5 and 17 years old. RESULTS: Overall, 36.4% of the children (39.8% of the boys and 32% of the girls) in the sample were in the combined category of at risk of overweight or overweight, representing an estimated 17 million US children. We found geographic variation at the state and the regional levels. The southeastern states, especially those west of the Appalachians and in the lower Mississippi region, had the highest prevalence of children who are at risk of overweight and/or overweight. The central Rocky Mountain states of Colorado, Utah, and Wyoming had the lowest prevalence, followed by the northwestern quadrant of the lower 48 states and New England. CONCLUSIONS: These National Survey of Children's Health data provide clinicians and public health professionals with useful data required for policy and planning related to childhood obesity at state levels. These data also serve as important baseline indicators and can be used to track changes over time. PMID- 17908725 TI - Weight faltering in infancy and IQ levels at 8 years in the Avon Longitudinal Study of Parents and Children. AB - OBJECTIVE: Our goal was to investigate the association between failure to thrive (defined as weight faltering in the first 9 months of life) and IQ levels 8 years later. METHODS: Weight gain (conditional on initial weight) from birth to 8 weeks, 8 weeks to 9 months, and birth to 9 months was measured on term infants from the Avon Longitudinal Study of Parents and Children. Cases of weight faltering were defined as those infants with a conditional weight gain below the 5th centile who were compared with the rest of the cohort as the control group. At the age of 8 years, 5771 infants born at term with no major congenital abnormalities had IQ measured by using the Wechsler Intelligence Scale for Children, Third Revision. RESULTS: Mean (SD) IQ scores were 104.7 (16.3) (total), 107.6 (16.5) (verbal), and 100.2 (16.9) (performance). Children whose weight faltered from birth to 9 months had a total IQ that was significantly lower by an average of -2.71 points at 8 years, equivalent to 0.17 SD. Weight gain from birth to 8 weeks had a positive linear association with child IQ at 8 years. This remained significant in a multivariate regression despite controlling for correlates of both infant growth and child IQ; 1 SD of weight gain was associated with a difference of 0.84 points in the total IQ score. In contrast to early weight faltering, weight gain from 8 weeks to 9 months was not related to IQ at 8 years. CONCLUSIONS: Failure to thrive in infancy was associated with persisting deficits in IQ at 8 years; the critical period for growth faltering was birth to 8 weeks. The relationship between infant growth from birth to 8 weeks and later intellectual development was approximately linear over the whole range of weight velocities. PMID- 17908726 TI - Healthy Buddies: a novel, peer-led health promotion program for the prevention of obesity and eating disorders in children in elementary school. AB - OBJECTIVE: We designed and tested a novel health promotion program for elementary schools that was based on peer teaching from older to younger schoolchildren ("Healthy Buddies"). SUBJECTS AND METHODS: This prospective pilot study compared the effect of our program (2-3 hours/week, 21 weeks) in 2 Canadian elementary schools (intervention: n = 232 children, the whole school implementing the program; control: n = 151). Older students (4th through 7th grade) were given direct instruction from 1 intervention teacher and were paired with younger students (kindergarten through 3rd grade) for the whole school year. Students in 4th through 7th grade then acted as teachers for their younger "buddies." All lessons included 3 components of healthy living: nutrition, physical activity, and healthy body image. The students first learned how to be positive buddies and learned the 3 components of a healthy life. Thereafter, they learned how to overcome challenges to living a healthy life. Outcome measures (intervention and control schools at the beginning and end of the school year) included validated questionnaires that assessed healthy-living knowledge, behavior and attitude, a 9 minute fitness run, self-competence, body satisfaction, disordered eating symptoms, and anthropometry (BMI, blood pressure, and heart rate). RESULTS: Compared with control students, both older and younger intervention students showed an increase in healthy-living knowledge, behavior, and attitude scores and a smaller increase in systolic blood pressure. BMI and weight increased less in the intervention students in 4th through 7th grade and height more in the intervention students in kindergarten through 3rd grade. CONCLUSIONS: Our student led curriculum improved knowledge not only in older schoolchildren but also in their younger buddies. It also decreased weight velocity in the older students. Student-led teaching may be an efficient, easy-to-implement way of promoting a healthy lifestyle from kindergarten to 7th grade. PMID- 17908727 TI - How do toddler eating problems relate to their eating behavior, food preferences, and growth? AB - OBJECTIVES: Eating problems are a common cause of concern for the parents of toddlers, but few studies have examined the correlates of eating problems or the growth patterns associated with them in a large population-based sample. Our goal was to examine the distribution of eating behaviors in a large representative sample of toddlers and their mothers' approach to feeding. In addition, we describe the prevalence of parentally perceived eating problems and how they relate to specific behaviors, food preferences, and growth in the child. METHODS: We conducted a cross-sectional analysis of data from a United Kingdom population based birth cohort, the Gateshead Millennium Baby Study, which included 455 questionnaires completed by parents when their children were aged 30 months. RESULTS: Eating was perceived to be a problem by 89 (20%) parents. Eating a limited variety (79 [17%]) and preferring drinks to food (57 [13%]) were the most prevalent problem behaviors. Thirty-seven children (8%) were described by parents as definitely "faddy" (picky), and these children liked fewer foods and had higher eating restriction scores than those described as not faddy. Children who were described as having an eating problem gained less weight over the first 2 years; 11.1% had weight faltering compared with 3.5% in children not described as having an eating problem. Being faddy was only weakly associated with poor growth, and simply eating a limited variety was unrelated to growth. High milk consumption was associated with lower appetite but not with poor growth. CONCLUSIONS: Eating problems are common in toddlers and in the majority are associated with normal growth, although weight faltering is more common in such children. Excessive milk-drinking may be a cause of low appetite at meal times. PMID- 17908728 TI - Down syndrome: a novel risk factor for respiratory syncytial virus bronchiolitis- a prospective birth-cohort study. AB - OBJECTIVES: Respiratory syncytial virus is the single-most important cause of lower respiratory tract infections in children. Preterm birth and congenital heart disease are known risk factors for severe respiratory syncytial virus infections. Although Down syndrome is associated with a high risk of respiratory tract infections, little is known about the incidence of respiratory syncytial virus infections in this group. The aim of our study was to determine the incidence of respiratory syncytial virus lower respiratory tract infection associated hospitalization among children with Down syndrome. PATIENTS AND METHODS: We performed a retrospective observational study and a prospective nationwide birth-cohort study of children with Down syndrome. The retrospective cohort comprised 176 children with Down syndrome. A birth cohort of 219 children with Down syndrome was prospectively followed until 2 years of age. All 276 siblings of the birth cohort were used as controls. RESULTS: Of the 395 patients with Down syndrome, 180 (45.6%) had a known risk factor for severe respiratory syncytial virus infections; 39 (9.9%) of these were hospitalized for respiratory syncytial virus lower respiratory tract infections. Two control children (0.7%) versus 9 term children with Down syndrome without congenital heart disease (7.6%) were hospitalized for respiratory syncytial virus lower respiratory tract infections. The median duration of hospitalization was 10 days; mechanical ventilation was required for 5 children (12.8%). CONCLUSIONS: This is the first study, to our knowledge, to demonstrate that Down syndrome is a novel independent risk factor for severe respiratory syncytial virus lower respiratory tract infections. These findings should prompt studies to investigate possible mechanisms that underlie severe respiratory syncytial virus lower respiratory tract infections in children with Down syndrome. The effect of respiratory syncytial virus prophylaxis in this specific population needs to be established. PMID- 17908730 TI - Swaddling: a systematic review. AB - Swaddling was an almost universal child-care practice before the 18th century. It is still tradition in certain parts of the Middle East and is gaining popularity in the United Kingdom, the United States, and The Netherlands to curb excessive crying. We have systematically reviewed all articles on swaddling to evaluate its possible benefits and disadvantages. In general, swaddled infants arouse less and sleep longer. Preterm infants have shown improved neuromuscular development, less physiologic distress, better motor organization, and more self-regulatory ability when they are swaddled. When compared with massage, excessively crying infants cried less when swaddled, and swaddling can soothe pain in infants. It is supportive in cases of neonatal abstinence syndrome and infants with neonatal cerebral lesions. It can be helpful in regulating temperature but can also cause hyperthermia when misapplied. Another possible adverse effect is an increased risk of the development of hip dysplasia, which is related to swaddling with the legs in extension and adduction. Although swaddling promotes the favorable supine position, the combination of swaddling with prone position increases the risk of sudden infant death syndrome, which makes it necessary to warn parents to stop swaddling if infants attempt to turn. There is some evidence that there is a higher risk of respiratory infections related to the tightness of swaddling. Furthermore, swaddling does not influence rickets onset or bone properties. Swaddling immediately after birth can cause delayed postnatal weight gain under certain conditions, but does not seem to influence breastfeeding parameters. PMID- 17908729 TI - Cutaneous reactions to drugs in children. AB - Cutaneous eruptions are a commonly reported adverse drug reaction. Cutaneous adverse drug reactions in the pediatric population have a significant impact on patients' current and future care options. A patient's recollection of having a "rash" when they took a medication as a child is a frequent reason for not prescribing a particular treatment. The quick detection and treatment of cutaneous adverse drug reactions, plus identification of the causative agent, are essential for preventing the progression of the reaction, preventing additional exposures, and ensuring the appropriate use of medications for both the current condition and others as the patient ages. The purpose of this review is to discuss a reasonable approach to recognition and initial management of cutaneous adverse drug reactions in children. PMID- 17908731 TI - Professionalism in pediatrics. AB - The purpose of this report is to provide a concrete overview of the ideal standards of behavior and professional practice to which pediatricians should aspire and by which students and residents can be evaluated. Recognizing that the ideal is not always achievable in the practical sense, this document details the key components of professionalism in pediatric practice with an emphasis on core professional values for which pediatricians should strive and that will serve as a moral compass needed to provide quality care for children and their families. PMID- 17908732 TI - Impact of Hurricane Katrina on newborn screening in Louisiana. AB - OBJECTIVE: The Louisiana Office of Public Health and the Centers for Disease Control and Prevention assessed the extent to which newborn screening was disrupted from August 15 to September 21, 2005, the immediate period before and after Hurricane Katrina. METHODS: A list of hospitals with labor and delivery services was obtained from the Louisiana Hospital Association. A survey sent to hospitals on October 17, 2005, asked about the number of live births during the assessment period, disruption in hospital services, the number of specimens sent to alternative laboratories, and the number of children without screening results. RESULTS: Among 64 Louisiana hospitals with labor and delivery units, 6 remained closed at the time of the survey. Of the 58 open hospitals, 53 (91.4%) completed the questionnaire. Twenty-one (36.2%) of 58 hospitals experienced disruption of newborn screening services. Respondents from 31 (58.5%) of the 53 open hospitals acknowledged receiving the advisory from the Office of Public Health regarding resumption of newborn screening laboratory services. Hospitals stated that of 5958 specimens submitted, reports had not been received for 1207 (20.3%) newborns. The Office of Public Health laboratory reviewed the names of 2828 newborns received from hospitals and determined that no specimen was received within 14 days of collection for 631 (22.3%). Thirty percent of the specimens received from infants who were born between August 15 and September 21 were rejected as a result of having been received >14 days after collection. Ten children had confirmed positive screening results during the assessment period; all were located, and treatment was initiated. CONCLUSIONS: Collaboration between the Office of Public Health and the Centers for Disease Control and Prevention was essential to increase awareness of changes in laboratory procedures after the hurricane and to help identify infants who might be in need of screening or rescreening. PMID- 17908734 TI - Sleep duration from ages 1 to 10 years: variability and stability in comparison with growth. AB - OBJECTIVE: Our goal was to describe the variability of sleep duration (time in bed per 24 hours) in healthy children from 1 to 10 years of age in comparison with growth measures. METHODS: A total of 305 children were followed with structured sleep-related interviews and measurements of height and weight 12, 18, and 24 months after birth and then at annual intervals until 10 years of age. SD scores were calculated, and smooth curves were fitted by smoothing splines through the SD scores. The long-term variability channel within children (units SD score) was defined as the difference between the maximum and the minimum of the smooth curves and the short-term variability channel (units SD score) as the difference of the largest and the smallest deviations of the original SD scores from the smooth curve. RESULTS: Sleep duration remained within a long-term variability channel <0.5 SD score in 21% of the children (34% for height, 21% for weight). Nearly every second child (46%) stayed within a long-term variability channel <1.0 SD score (76% for height, 64% for weight). Sleep duration of approximately 90% of all children ran within a long-term variability channel of <2.0 SD score (corresponding, eg, to the range between the 2nd and the 50th percentile). No single child's sleep duration remained within a short-term variability channel <0.5 SD score, indicating fluctuations from year to year (60% for height, 53% for weight). An association between aspects of sleep duration and somatic growth was not observed at any age. CONCLUSIONS: Sleep duration during early and middle childhood shows large variability among children, as well as trait-like long-term stability and state-like yearly fluctuations within children. An individual approach to the child's sleep behavior is needed; expectations in terms of appropriate sleep duration of the child should be adjusted to the individual sleep need. PMID- 17908733 TI - Prehospital preparedness for pediatric mass-casualty events. AB - OBJECTIVES: Recent events have reiterated the need for well-coordinated planning for mass-casualty events, including those that involve children. The objective of this study was to document the preparedness of prehospital emergency medical services agencies in the United States for the care of children who are involved in mass-casualty events. METHODS: A national list of all licensed prehospital emergency medical services agencies was prepared through contact with each state's emergency medical services office. A survey was mailed to 3748 emergency medical services agencies that were selected randomly from the national list in November 2004; a second survey was mailed to nonresponders in March 2005. Descriptive statistics were used to describe study variables. RESULTS: Most (72.9%) agencies reported having a written plan for response to a mass-casualty event, but only 248 (13.3%) reported having pediatric-specific mass-casualty event plans. Most (69%) services reported that they did not have a specific plan for response to a mass-casualty event at a school. Most (62.1%) agencies reported that their mass-casualty event plan does not include provisions for people with special health care needs. Only 19.2% of the services reported using a pediatric specific triage protocol for mass-casualty events, and 12.3% reported having a pediatrician involved in their medical control. Although most (69.3%) agencies reported participation in a local or regional disaster drill in the past year, fewer than half of those that participated in drills (49.0%) included pediatric victims. CONCLUSIONS: Although children are among the most vulnerable in the event of disaster, there are substantial deficiencies in the preparedness plans of prehospital emergency medical services agencies in the United States for the care of children in a mass-casualty event. PMID- 17908735 TI - Lumbar puncture success rate is not influenced by family-member presence. AB - OVERVIEW: The presence of a family member during invasive pediatric procedures such as lumbar puncture has been shown to reduce patient anxiety. However, family presence might also affect clinicians' stress and anxiety, with uncertain consequences for procedural success. OBJECTIVE: Our goal was to evaluate the association between family-member presence and lumbar puncture success rates. DESIGN/METHODS: We performed a prospective cohort study of all children who underwent a lumbar puncture in a single pediatric emergency department between July 2003 and January 2005. The presence of a family member was documented by the physician who performed the lumbar puncture. Success rates were assessed by using 2 main outcomes: (1) the rate of traumatic (cerebrospinal fluid red blood cells > or = 10,000 cells per microL) or unsuccessful lumbar puncture (no cerebrospinal fluid sent for cell counts) and (2) the number of lumbar puncture attempts. Multivariate analyses were adjusted for patient age, race, time of day, physician experience, use of local anesthetic, catheter stylet removal, and patient movement during the procedure. RESULTS: Of the 1474 eligible lumbar punctures, 1459 (99%) were included in the analysis. A family member was present for 1178 (81%) of the procedures studied. A total of 1267 (87%) lumbar punctures were nontraumatic, and 192 (13%) were traumatic or unsuccessful. Neither the rate of traumatic or unobtainable lumbar punctures nor the number of lumbar puncture attempts differed based on whether a family member was present for the procedure. CONCLUSIONS: The presence of a family member was not associated with an increased risk of traumatic or unobtainable lumbar puncture, nor was it associated with more attempts at the procedure. The benefits of having a family member present during the procedure were not counterbalanced by adverse effects on procedural success. PMID- 17908736 TI - Validity of parental report of influenza vaccination in children 6 to 59 months of age. AB - OBJECTIVE: The objective of this study was to assess the validity of parental report of influenza vaccinations. PATIENTS AND METHODS: A cross-sectional study of children who were 6 to 59 months of age and presented to a large, pediatric residency clinic from February through April 2005 was performed. A standardized, parental questionnaire ascertained the influenza vaccination status of children during the 2004-2005 influenza season and was compared with the medical chart, the criterion standard. Children were classified as being at high risk when they had a specific influenza vaccine recommendation in 2004-2005 by age (6-23 months of age) or by chronic medical condition. RESULTS: Of 218 parents approached in the pediatric residency clinic, 198 (95%) children who were 6 to 59 months of age were enrolled, and 84 (42%) were vaccinated according to the medical chart. More children who were 6 to 23 months than those who were 24 to 59 months of age were vaccinated (63% vs 21%). Children with chronic medical conditions were more likely to be vaccinated than healthy children who were 24 to 59 months of age (57% vs 11%), but no difference was observed for children who were 6 to 23 months of age (79% vs 60%). In comparison with the medical chart, parental report of influenza vaccination had a sensitivity of 88%, a specificity of 90%, and a kappa coefficient of 0.78. For children who were 6 to 23 months of age or had a chronic medical condition (n = 123), parental report had a sensitivity of 89%, a specificity of 81%, and a kappa coefficient of 0.71. CONCLUSIONS: Parental report of influenza vaccination among children who were 6 to 59 months of age had reasonable sensitivity, specificity, and reliability as compared with the medical chart in this study population. PMID- 17908737 TI - Postdischarge infant mortality among very low birth weight infants: a population based study. AB - OBJECTIVE: The objective of this study was to identify factors that were associated with death after discharge from the NICU of very low birth weight infants in a population-based study. METHODS: From a national cohort of 13,430 very low birth weight infants who were born in Israel from 1995 to 2003, 10,602 infants were discharged from the hospital and composed the study population. Demographic and clinical data regarding the pregnancy, delivery, and neonatal course were obtained from the Israel national very low birth weight infant database. Data on each case of death during the postdischarge period until 1 year of age were provided by the Ministry of Health from national linked birth and death certificates. Univariate analyses and a multivariable logistic regression analyses were performed to examine the perinatal and neonatal risk factors for postdischarge death. RESULTS: The postdischarge mortality rate was 7.5 per 1000 (80 of 10,602 infants discharged from the hospital). The death rate was significantly higher in non-Jewish infants, infants who were born to young mothers, and infants who were born to low-educated mothers. After adjustment for demographic characteristics and perinatal and neonatal variables, postdischarge mortality was independently associated with congenital malformations, neonatal seizures, necrotizing enterocolitis, and bronchopulmonary dysplasia. CONCLUSION: Although the postdischarge death rate was relatively low in our cohort of very low birth weight infants, attention should be focused on the subgroups of infants who are at higher risk to decrease their mortality further. PMID- 17908738 TI - Determinants of child-parent agreement in quality-of-life reports: a European study of children with cerebral palsy. AB - OBJECTIVES: The differences between child self-reports and parent proxy reports of quality of life in a large population of children with cerebral palsy were studied. We examined whether child characteristics, severity of impairment, socioeconomic factors, and parental stress were associated with parent proxy reports being respectively higher or lower than child self-reports of quality of life. METHODS: This study was conducted in 2004-2005 and assessed child quality of life (using the Kidscreen questionnaire, 10 domains, each scored 0-100) through self-reports and parent proxy reports of 500 children aged 8 to 12 years who had cerebral palsy and were living in 7 countries in Europe. RESULTS: The mean child-reported scores of quality of life were significantly higher than the parent proxy reports in 8 domains, significantly lower for the finances domain, and similar for the emotions domain. The average frequency of disagreement (child parent difference greater than half an SD of child scores) over all domains was 64%, with parents rating their child's quality of life lower than the children themselves in 29% to 57% of child-parent pairs. We found that high levels of stress in parenting negatively influenced parents' perception of their child's quality of life, whereas the main factor explaining parents' ratings of children's quality of life higher than the children themselves is self-reported severe child pain. CONCLUSIONS: This study shows that the factors associated with disagreement are different according to the direction of disagreement. In particular, parental well-being and child pain should be taken into account in the interpretation of parent proxy reports, especially when no child self-report of quality of life is available. In the latter cases, it may be advisable to obtain additional proxy reports (from caregivers, teachers, or clinicians) to obtain complementary information on the child's quality of life. PMID- 17908739 TI - Characteristics of neonatal units that care for very preterm infants in Europe: results from the MOSAIC study. AB - OBJECTIVES: We sought to compare guidelines for level III units in 10 European regions and analyze the characteristics of neonatal units that care for very preterm infants. METHODS: The MOSAIC (Models of Organising Access to Intensive Care for Very Preterm Births) project combined a prospective cohort study on all births between 22 and 31 completed weeks of gestation in 10 European regions and a survey of neonatal unit characteristics. Units that admitted > or = 5 infants at < 32 weeks of gestation were included in the analysis (N = 111). Place of hospitalization of infants who were admitted to neonatal care was analyzed by using the cohort data (N = 4947). National or regional guidelines for level III units were reviewed. RESULTS: Six of 9 guidelines for level III units included minimum size criteria, based on number of intensive care beds (6 guidelines), neonatal admissions (2), ventilated patients (1), obstetric intensive care beds (1), and deliveries (2). The characteristics of level III units varied, and many were small or unspecialized by recommended criteria: 36% had fewer than 50 very preterm annual admissions, 22% ventilated fewer than 50 infants annually, and 28% had fewer than 6 intensive care beds. Level II units were less specialized, but some provided mechanical ventilation (57%) or high-frequency ventilation (20%) or had neonatal surgery facilities (17%). Sixty-nine percent of level III and 36% of level I or II units had continuous medical coverage by a qualified pediatrician. Twenty-two percent of infants who were < 28 weeks of gestation were treated in units that admitted fewer than 50 very preterm infants annually (range: 2%-54% across the study regions). CONCLUSIONS: No consensus exists in Europe about size or other criteria for NICUs. A better understanding of the characteristics associated with high-quality neonatal care is needed, given the high proportion of very preterm infants who are cared for in units that are considered small or less specialized by many recommendations. PMID- 17908740 TI - Effects of nurse home visiting on maternal and child functioning: age-9 follow-up of a randomized trial. AB - OBJECTIVE: Our goal was to test the effect of prenatal and infancy home visits by nurses on mothers' fertility and children's functioning 7 years after the program ended at child age 2. METHODS: We conducted a randomized, controlled trial in a public system of obstetric and pediatric care. A total of 743 primarily black women <29 weeks' gestation, with previous live births and at least 2 sociodemographic risk characteristics (unmarried, <12 years of education, unemployed), were randomly assigned to receive nurse home visits or comparison services. Primary outcomes consisted of intervals between births of first and second children and number of children born per year; mothers' stability of relationships with partners and relationships with the biological father of the child; mothers' use of welfare, food stamps, and Medicaid; mothers' use of substances; mothers' arrests and incarcerations; and children's academic achievement, school conduct, and mental disorders. Secondary outcomes were the sequelae of subsequent pregnancies, women's employment, experience of domestic violence, and children's mortality. RESULTS: Nurse-visited women had longer intervals between births of first and second children, fewer cumulative subsequent births per year, and longer relationships with current partners. From birth through child age 9, nurse-visited women used welfare and food stamps for fewer months. Nurse-visited children born to mothers with low psychological resources, compared with control-group counterparts, had better grade-point averages and achievement test scores in math and reading in grades 1 through 3. Nurse-visited children, as a trend, were less likely to die from birth through age 9, an effect accounted for by deaths that were attributable to potentially preventable causes. CONCLUSIONS: By child age 9, the program reduced women's rates of subsequent births, increased the intervals between the births of first and second children, increased the stability of their relationships with partners, facilitated children's academic adjustment to elementary school, and seems to have reduced childhood mortality from preventable causes. PMID- 17908741 TI - Effect of vitamin A and zinc supplementation on gastrointestinal parasitic infections among Mexican children. AB - OBJECTIVE: Gastrointestinal parasites continue to be an important cause of morbidity and stunting among children in developing countries. We evaluated the effect of vitamin A and zinc supplementation on infections by Giardia lamblia, Ascaris lumbricoides, and Entamoeba histolytica. METHODS: A randomized, double blind, placebo-controlled trial was conducted among 707 children who were 6 to 15 months of age and from periurban areas of Mexico City, Mexico, between January 2000 and May 2002. Children, who were assigned to receive either vitamin A every 2 months, a daily zinc supplement, a combined vitamin A and zinc supplement, or a placebo, were followed for 1 year. The primary end points were the 12-month rates and durations of infection for the 3 parasites and rates of parasite-associated diarrheal disease as determined in stools collected once a month and after diarrheal episodes. RESULTS: G. lamblia infections were reduced and A. lumbricoides infections increased among children in the combined vitamin A and zinc group or the zinc alone group, respectively. Durations of Giardia infections were reduced among children in all 3 treatment arms, whereas Ascaris infections were reduced in the vitamin A and zinc group. In contrast, E. histolytica infection durations were longer among zinc-supplemented children. Finally, E. histolytica- and A. lumbricoides-associated diarrheal episodes were reduced among children who received zinc alone or a combined vitamin A and zinc supplement, respectively. CONCLUSIONS: We found that vitamin A and zinc supplementation was associated with distinct parasite-specific health outcomes. Vitamin A plus zinc reduces G. lamblia incidence, whereas zinc supplementation increases A. lumbricoides incidence but decreases E. histolytica-associated diarrhea. PMID- 17908742 TI - Effectiveness of previous mumps vaccination during a summer camp outbreak. AB - OBJECTIVES: Mumps is a vaccine-preventable disease that may cause outbreaks. In July 2005, an outbreak of mumps occurred during a children's summer camp in upstate New York. An investigation was initiated to describe the cases and evaluate vaccine effectiveness. METHODS: A retrospective cohort study was conducted among 541 children from the United States and abroad who attended a 1- or 2-month overnight summer camp. Patients with mumps were interviewed; serologic analysis was conducted for 6 case patients. Vaccine effectiveness was calculated by retrospective review of immunization records for 507 attendees who were eligible for vaccination and had verified immunization history. RESULTS: Thirty one camp attendees were identified as having mumps (attack rate: 5.7%); 5 (83%) of 6 patients tested had positivity for mumps immunoglobulin M. Of the 507 participants (including 29 patients) with available immunization history, 440 (including 16 [87%] patients) were 2-dose recipients of mumps vaccine (attack rate: 3.6%); 46 participants (including 4 [9%] patients) were 1-dose recipients (attack rate: 8.7%); and 21 (including 9 [4%] patients) were unvaccinated (attack rate: 42.9%). Vaccine effectiveness was 92% for 2 doses and 80% for 1 dose. CONCLUSIONS: Outbreaks of mumps in settings such as summer camps can occur despite high vaccination rates. Vaccine effectiveness for 2 mumps vaccinations was greater than vaccine effectiveness for 1 mumps vaccination. Therefore, recommendation of 2 mumps vaccinations for summer camp participants continues to be appropriate. Control of mumps disease relies on broad vaccination coupled with correct clinical diagnosis and strict control measures. PMID- 17908743 TI - Small changes in dietary sugar and physical activity as an approach to preventing excessive weight gain: the America on the Move family study. AB - OBJECTIVES: The intent of this study was to evaluate whether small changes in diet and physical activity, as promoted by the America on the Move initiative, could prevent excessive weight gain in overweight children. METHODS: In this family-intervention study, the America on the Move small-changes approach for weight-gain prevention was evaluated in families with at least 1 child (7-14 years old) who was overweight or at risk for overweight. These children were the primary target of the intervention, and parents were the secondary target. Families were randomly assigned to either the America on the Move group (n = 100) or the self-monitor-only group (n = 92). Families who were assigned to the America on the Move group were asked to make 2 small lifestyle changes: (1) to walk an additional 2000 steps per day above baseline as measured by pedometers and (2) to eliminate 420 kJ/day (100 kcal/day) from their typical diet by replacing dietary sugar with a noncaloric sweetener. Families who were assigned to the self-monitor group were asked to use pedometers to record physical activity but were not asked to change their diet or physical activity level. RESULTS: During a 6-month period, both groups of children showed significant decreases in BMI for age. However, the America on the Move group compared with the self-monitor group had a significantly higher percentage of target children who maintained or reduced their BMI for age and, consistently, a significantly lower percentage who increased their BMI for age. There was no significant weight gain during the 6-month intervention in parents of either group. CONCLUSIONS: The small-changes approach advocated by America on the Move could be useful for addressing childhood obesity by preventing excess weight gain in families. PMID- 17908744 TI - Validation of the Gross Motor Function Measure for use in children and adolescents with traumatic brain injuries. AB - OBJECTIVES: Motor function recovery is a key goal during rehabilitation of children and adolescents with traumatic brain injury. To evaluate how well treatment strategies improve motor function, we need validated outcome measures that are responsive to change in pediatric patients with traumatic brain injury. The Gross Motor Function Measure has demonstrated excellent psychometric properties in children with cerebral palsy and Down syndrome, yet its responsiveness in patients with pediatric traumatic brain injury has not been proven irrefutably. Our aim was to validate the Gross Motor Function Measure for this patient group. METHODS: Seventy-three patients (mean age: 11.4 years; range: 0.8-18.9 years) with moderate-to-severe traumatic brain injury were recruited in 12 rehabilitation centers and assessed twice with the Gross Motor Function Measure-88 over 4 to 6 weeks. As an external standard, we used judgements of change made independently by parents, physiotherapists, and 2 video assessors who were not familiar with the patients. We formulated and statistically investigated a priori hypotheses of how Gross Motor Function Measure change scores would correlate with those judgements of change. Both Gross Motor Function Measure versions, the original Gross Motor Function Measure-88 and the more recently developed Gross Motor Function Measure-66, were evaluated. RESULTS: Both Gross Motor Function Measure change scores correlated significantly with all of the clinical judgements of change. The degree of correlation that we postulated, that the Gross Motor Function Measure change score would correlate highest with the video rating followed by physiotherapists and parents, was fully confirmed by the Gross Motor Function Measure-88 and largely confirmed by the Gross Motor Function Measure-66. Both Gross Motor Function Measure versions revealed convincing discriminative capability. Test-retest reliability was excellent. CONCLUSIONS: We demonstrate convincing evidence of responsiveness and validity to support the use of both Gross Motor Function Measure versions as evaluative measures of gross motor function in children and adolescents with traumatic brain injury. PMID- 17908745 TI - Family-member presence during interventions in the intensive care unit: perceptions of pediatric cardiac intensive care providers. AB - OBJECTIVE: Should family members be present during interventions in an ICU? This question is a source of debate among health care providers. We propose to define perceptions and practice regarding family-member presence during ICU interventions from a multidisciplinary group of pediatric cardiac intensive care providers. METHODS: A 20-question survey was created and distributed to attendees of the 2004 Pediatric Cardiac Intensive Care Symposium, 1 year after the meeting. Interventions were defined as noninvasive (team rounds), invasive (tracheal intubation, central/arterial line placement, chest tube placement, or pericardiocentesis), or extremely invasive (cardiopulmonary resuscitation). RESULTS: A total of 211 surveys (145 physicians and 66 nonphysicians) were completed. Of all responders, the majority believe family members have a right to be present during cardiopulmonary resuscitation (75%), team rounds (77%), and invasive procedures (57%). Sixty-five percent of respondents encounter families that frequently request to be present for team rounds. However, the majority of respondents encounter families that rarely request to be present during invasive procedures (69%) and cardiopulmonary resuscitation (73%). Many providers practice in ICUs where family-member presence is allowed; 64% allow family members to attend team rounds. Some of the concerns providers have regarding family-member presence in the ICU include family-member presence causing stress to the provider during invasive procedures along with distractions and nervousness among the team during cardiopulmonary resuscitation. The majority of providers predict family member presence during cardiopulmonary resuscitation would not increase medicolegal concerns. CONCLUSIONS: Most respondents, nonphysicians more than physicians, believe that family members have a right to be present during all ICU interventions. The majority of respondents encounter families that frequently request to be present for team rounds. However, the majority of respondents encounter families that rarely request to be present during invasive procedures and cardiopulmonary resuscitation. Most respondents believe family-member presence during cardiopulmonary resuscitation would not increase medicolegal concerns. PMID- 17908746 TI - Improving asthma-related health outcomes among low-income, multiethnic, school aged children: results of a demonstration project that combined continuous quality improvement and community health worker strategies. AB - OBJECTIVE: The purpose of this work was to improve asthma-related health outcomes in an ethnically and geographically disparate population of economically disadvantaged school-aged children by using a team-based approach using continuous quality improvement and community health workers. PATIENTS AND METHODS: A demonstration project was conducted with 7 community clinics treating approximately 3000 children with asthma 5 to 18 years of age. The overall clinic population with asthma was assessed for care-process changes through random cross sectional chart reviews at baseline and 24 months (N = 560). A subset of patients with either moderate or severe persistent asthma or poorly controlled asthma (N = 405) was followed longitudinally for specific asthma-related clinical outcomes, satisfaction with care, and confidence managing asthma by family interview at baseline and at 12 or 24 months. Patient-centered and care-process outcomes included patient/parent assessment of quality of care and confidence in self management, asthma action plan review, and documentation of guideline-based indicators of quality of care. Direct clinical outcomes included daytime and nighttime symptoms, use of rescue medications, acute care and emergency department visits, hospitalizations, and missed school days. Each clinic site's degree of adherence to the intervention model was evaluated and ranked to examine the correlation between model adherence and outcomes. RESULTS: Cross-sectional data showed clinic-wide improvements in the documentation of asthma severity, review of action plans, health services use, and asthma symptoms. At follow-up in the longitudinal sample, fewer patients reported acute visits, emergency department visits, hospitalizations, frequent daytime and nighttime symptoms, and missed school days compared with baseline. More patients reported excellent or very good quality of care and confidence in asthma self-management. Linear regression analysis of the clinical sites' model adherence ranks against site level combined scores estimating overall outcomes, clinical outcomes, and improvements in clinical care processes showed significant linear correlations with R2 > or = 0.60. CONCLUSIONS: The demonstration produced major improvements in asthma-related care processes and clinical outcomes. Closer adherence to the demonstration model was directly associated with better outcomes. PMID- 17908747 TI - Immune function in young children with previous pulmonary or miliary/meningeal tuberculosis and impact of BCG vaccination. AB - OBJECTIVE: Children <5 years old are at increased risk of miliary/meningeal tuberculosis, but the immunologic factors that place them at risk are unknown. BCG vaccine protects against miliary/meningeal tuberculosis, but the mechanism of protection is unknown. We assessed for abnormalities in immune response associated with miliary/meningeal or pulmonary tuberculosis in young children. PATIENTS AND METHODS: We conducted a case-control study among HIV-seronegative Brazilian children who were <5 years old. Case subjects had previous culture confirmed or clinical miliary/meningeal tuberculosis. There were 2 sets of control subjects: those with culture-confirmed pulmonary tuberculosis and purified protein derivative-positive household contacts. All of the children had completed treatment. Peripheral blood mononuclear cells were stimulated (phytohemagglutinin, phytohemagglutinin + interleukin 12, lipopolysaccharide, lipopolysaccharide + interferon-gamma, and purified protein derivative), and cytokine responses (interleukin 1beta, interleukin-4, interleukin-6, interleukin 8, interleukin 10, interleukin 12, interferon-gamma, tumor necrosis factor-alpha, and monocyte chemoattractant protein 1) were quantified by bead-based assay. Median cytokine responses were compared by the Kruskal-Wallis test. Multivariate analysis of variance accounted for multiple comparisons. RESULTS: There were 18 case subjects with miliary/meningeal tuberculosis, 28 pulmonary control subjects, and 29 purified protein derivative-positive control subjects. The median age was 4.2 years. There was no difference in case and control subjects by age, gender, race, BMI, or median CD4 count. Twelve (67%) of 18 case subjects, 26 (93%) of 28 pulmonary control subjects, and 28 (97%) of 29 purified protein derivative positive subjects had received BCG vaccine. No cytokine defects were identified in case subjects with miliary/meningeal tuberculosis compared with either set of control subjects. Pulmonary control subjects had uniformly higher monocyte chemoattractant protein 1 levels than case subjects with miliary/meningeal tuberculosis and purified protein derivative-positive control subjects, both at rest and with lipopolysaccharide, lipopolysaccharide + interferon-gamma, and purified protein derivative stimulation. Pulmonary control subjects did not have a higher frequency of allele G in the -2518 monocyte chemoattractant protein 1 promoter polymorphism. Case subjects with miliary/meningeal tuberculosis who had received BCG vaccine (n = 12) had lower stimulated interleukin 8 production than children who did not receive BCG vaccine (n = 6). CONCLUSIONS: Children with previous miliary/meningeal tuberculosis did not have a major defect in the cytokine pathways studied. Increased monocyte chemoattractant protein 1 levels were associated with pulmonary disease, occurred despite BCG vaccination, and were not associated with a polymorphism in the monocyte chemoattractant protein 1 promoter. PMID- 17908748 TI - Myocardial infarction in healthy adolescents. AB - OBJECTIVE: Chest pain in children and adolescents is a frequent cause for office or emergency department visits. However, it is unclear whether myocardial infarction occurs in children with no anatomic abnormality presenting with chest pain. METHODS: Clinical history, electrocardiography, echocardiography, and cardiac enzyme levels were evaluated in patients presenting to the emergency department over a period of 11 years (June 1995 to May 2006). Patients in whom findings were suggestive of acute myocardial infarction, in addition, underwent drug screening, serum lipid profile, and hypercoagulability workup and, when myocardial infarction was diagnosed, heart catheterization with coronary angiography. RESULTS: Nine patients (8 boys; age range: 12-20 years; mean: 15.5 years) met established criteria for myocardial infarction. Abnormal electrocardiograms were found in 8 patients (6 with ST elevation and 2 with nonspecific ST-T abnormalities), abnormal cardiac enzyme levels in all, and echocardiographic abnormalities in 3. Cardiac dysrhythmias were found in 4 patients, 3 with nonsustained ventricular tachycardia. Drug abuse, lipid profile, and hypercoagulability studies were negative in all. Left ventricular focal hypokinesia was seen by echocardiogram or angiography in 5 patients and abnormal coronary anatomy in none. Cardiac function normalized in 8 patients. One patient had a persistent focal inferior hypokinesis. Calcium channel blocker therapy was initiated in all of the patients with no recurrence of anginal chest pain on follow-up. One patient complained of chest pain distinct from anginal pain. CONCLUSIONS: Myocardial infarction can occur in adolescents with normal coronary arterial anatomy. Adolescents who present for emergency care with typical chest pain need electrocardiographic and cardiac enzyme workups. Those with results that are suggestive of acute infarction require additional workup. Coronary vasodilation therapy seems helpful, but given the lack of coronary thrombosis in these patients, thrombolytic therapy seems unwarranted. Long-term follow-up is necessary, and adjustments in therapy may be required with time. PMID- 17908749 TI - Association between infant breastfeeding and early childhood caries in the United States. AB - OBJECTIVE: Despite limited epidemiologic evidence, concern has been raised that breastfeeding and its duration may increase the risk of early childhood caries. The objective of this study was to assess the potential association of breastfeeding and other factors with the risk for early childhood caries among young children in the United States. METHODS: Data about oral health, infant feeding, and other child and family characteristics among children 2 to 5 years of age (N = 1576) were extracted from the 1999-2002 National Health and Nutrition Examination Survey. The association of breastfeeding and its duration, as well as other factors that previous research has found associated with early childhood caries, was examined in bivariate analyses and by multivariable logistic and Poisson regression analyses. RESULTS: After adjusting for potential confounders significant in bivariate analyses, breastfeeding and its duration were not associated with the risk for early childhood caries. Independent associations with increased risk for early childhood caries were older child age, poverty, being Mexican American, a dental visit within the last year, and maternal prenatal smoking. Poverty and being Mexican American also were independently associated with severe early childhood caries, whereas characteristics that were independently associated with greater decayed and filled surfaces on primary teeth surfaces were poverty, a dental visit within the last year, 5 years of age, and maternal smoking. CONCLUSIONS: These data provide no evidence to suggest that breastfeeding or its duration are independent risk factors for early childhood caries, severe early childhood caries, or decayed and filled surfaces on primary teeth. In contrast, they identify poverty, Mexican American ethnic status, and maternal smoking as independent risk factors for early childhood caries, which highlights the need to target poor and Mexican American children and those whose mothers smoke for early preventive dental visits. PMID- 17908750 TI - Persistent beneficial effects of breast milk ingested in the neonatal intensive care unit on outcomes of extremely low birth weight infants at 30 months of age. AB - BACKGROUND: We previously reported beneficial effects of breast milk ingestion by infants with extremely low birth weight in the NICU on developmental outcomes at 18 months' corrected age. The objective of this study was to determine whether these effects of breast milk in infants with extremely low birth weight persisted at 30 months' corrected age. METHODS: Nutrition data, including enteral and parenteral feeds, were prospectively collected, and 30 months' corrected age follow-up assessments were completed on 773 infants with extremely low birth weight who participated in the National Institute of Child Health and Human Development Neonatal Research Network Glutamine Trial. A total of 593 ingested some breast milk during the neonatal hospitalization, and 180 ingested none. Neonatal feeding characteristics and morbidities and 30-month interim history, neurodevelopmental outcomes, and growth parameters were analyzed. Children were divided into quintiles of breast milk volume to evaluate the effects of volume of human milk ingested during the NICU hospitalization. RESULTS: At 30 months, increased ingestion of breast milk was associated with higher Bayley Mental Developmental Index scores, higher Bayley behavior score percentiles for emotional regulation, and fewer rehospitalizations between discharge and 30 months. There were no differences in growth parameters or cerebral palsy. For every 10 mL/kg per day increase in breast milk, the Mental Developmental Index increased by 0.59 points, the Psychomotor Developmental Index by 0.56 points, and the total behavior percentile score by 0.99 points, and the risk of rehospitalization between discharge and 30 months decreased by 5%. CONCLUSIONS: Beneficial effects of ingestion of breast milk in the NICU persist at 30 months' corrected age in this vulnerable extremely low birth weight population. Continued efforts must be made to offer breast milk to all extremely low birth weight infants both in the NICU and after discharge. PMID- 17908751 TI - Trends in pediatric organ donation after cardiac death. AB - OBJECTIVE: Organ donation after cardiac death is viewed as one way of partially closing the current gap between organ supply and demand. There are no published guidelines for organ donation after cardiac death specific to the pediatric population. The objective of this study was to examine the cumulative pediatric donation-after-cardiac-death experience to set the context for the development and sharing of best-practice guidelines. PATIENTS AND METHODS: This was a retrospective, descriptive study that used data from the Organ Procurement and Transplantation Network/United Network for Organ Sharing database from 1993 to 2005. Organ data from all donors after cardiac death who were < 18 years of age were analyzed. The list of donor medical centers was then cross-referenced with the member list from the National Association of Children's Hospitals and Related Institutions. RESULTS: There were 683 organs from donation-after-cardiac-death donors < 18 years of age. Of those, < 5% were used for pediatric recipients. In comparison, approximately 20% of non-donation-after-cardiac-death organs from pediatric donors were used for pediatric recipients. The vast majority of donation-after-cardiac-death organs donated were kidneys and livers. More than 50% of medical centers that had a pediatric organ-donation-after-cardiac-death donor had just 1. The medical center with the largest pediatric organ-donation after-cardiac-death donation experience had 14 donors. Forty-three percent of medical centers that had > or = 1 pediatric donation-after-cardiac-death donor were members of the National Association of Children's Hospitals and Related Institutions. Fifty-six percent of all of the pediatric donation-after-cardiac death organs were donated from the National Association of Children's Hospitals and Related Institution member centers. CONCLUSIONS: Data regarding the use of pediatric donation-after-cardiac-death organs for pediatric recipients remain sparse. Few medical centers have had enough donation-after-cardiac-death donor experience to report a tried-and-true approach. We advocate for comprehensive collection and reporting of outcome data for all-aged recipients of pediatric donation-after-cardiac-death organs to help facilitate the generation of evidence based best-practice guidelines for pediatric donation after cardiac death. PMID- 17908752 TI - Contribution of inherited heart disease to sudden cardiac death in childhood. AB - BACKGROUND: In children aged 1 to 18 years, the causes of sudden cardiac death may remain unresolved when autopsy results are negative. Because inherited cardiac diseases are likely, cardiologic and genetic investigations of relatives may still yield the diagnosis in these cases. Moreover, these investigations provide timely identification of relatives who are also at risk of sudden cardiac death. We aimed to establish the cause of sudden cardiac death in the children of whom the family was referred to our cardiogenetics department and the diagnostic yield of these investigations. METHODS AND RESULTS: We genetically counseled 25 consecutive, unrelated families after sudden cardiac death of a child (aged 1 to 18 years) who was disease-free during lifetime and in whose family there was no known inherited heart disease. We performed cardiac investigation (electrocardiography, exercise testing, and echocardiography) of first-degree and second-degree relatives and performed diagnosis-directed DNA analysis. Autopsy was performed in 20 case subjects. A diagnosis was identified in 14 of 25 families. In addition, we studied 10 children after aborted sudden cardiac death; in 6 of them, a diagnosis was made. Overall, in 17 of the 19 families in whom an inherited disease was diagnosed, a disease-causing mutation in either a first degree relative or the index patient confirmed the diagnosis. CONCLUSIONS: Sudden cardiac death in children seems to be caused often by inherited cardiac diseases. Cardiac and genetic examination of relatives combined, if possible, with postmortem analysis after sudden cardiac death of a child has a high diagnostic yield (14 of 25), comparable to analysis in surviving victims of sudden cardiac death (6 of 10). Because sudden cardiac death can be prevented by timely treatment, these results warrant active family screening after unexplained sudden cardiac death of a child. PMID- 17908753 TI - Susceptibility to nicotine dependence: the Development and Assessment of Nicotine Dependence in Youth 2 study. AB - OBJECTIVES: The purpose of this work was to identify characteristics that predict progression from the first inhalation of a cigarette to dependence. We studied a cohort of 1246 public school 6th-graders in 6 Massachusetts communities (mean age at baseline: 12.2 years). METHODS: We conducted a 4-year prospective study using 11 interviews. We assessed 45 risk factors and measured diminished autonomy over tobacco with the Hooked on Nicotine Checklist and evaluated tobacco dependence according to the International Classification of Diseases, 10th Revision. Cox proportional-hazards models were used. RESULTS: Among 217 youths who had inhaled from a cigarette, the loss of autonomy over tobacco was predicted by feeling relaxed the first time inhaling from a cigarette and depressed mood. Tobacco dependence was predicted by feeling relaxed, familiarity with Joe Camel, novelty seeking, and depressed mood. CONCLUSIONS: Once exposure to nicotine had occurred, remarkably few risk factors for smoking consistently contributed to individual differences in susceptibility to the development of dependence or loss of autonomy. An experience of relaxation in response to the first dose of nicotine was the strongest predictor of both dependence and lost autonomy. This association was not explained by trait anxiety or any of the other measured psychosocial factors. These results are discussed in relation to the theory that the process of dependence is initiated by the first dose of nicotine. PMID- 17908754 TI - Family breakup and adolescents' psychosocial maladjustment: public health implications of family disruptions. AB - OBJECTIVES: Recent changes in family structure are associated with an increase in psychosocial maladjustment in adolescents. We examined, from a public health intervention perspective, the association between family breakup and psychosocial maladjustment in adolescents and assessed the mediating role of family functioning variables. METHODS: We analyzed data from the Social and Health Survey of Children and Adolescents in Quebec, Montreal, Canada, which was conducted in 1999. Sample-weighted logistic regression analyses were performed to determine the risk of internalizing disorders, externalizing disorders, substance abuse, and alcohol consumption in relation to family breakups and family functioning variables, after adjusting for socioeconomic factors. RESULTS: All 4 of the indicators of psychosocial maladjustment were significantly associated with family breakup. The association between family breakups and internalizing disorders was mediated by parental psychological distress and low paternal emotional support. Independently, the witnessing of interparental violence was also strongly associated with internalizing disorders. For the other 3 outcomes, that is, externalizing disorders, substance abuse, and alcohol consumption, family breakup and family-functioning variables had independent effects. CONCLUSIONS: Family-based interventions and social approaches are complementary support modalities for adolescents experiencing family disruptions. PMID- 17908755 TI - Patient-physician e-mail: an opportunity to transform pediatric health care delivery. AB - OBJECTIVE: The objectives of this study were to assess the patterns of patients who use a patient-physician e-mail service, measure physician time required to answer a patient question via e-mail compared with that via telephone, and determine the satisfaction of families who are provided e-mail access to their child's rheumatologist. METHODS: A consecutive series of patients' families were offered e-mail access during a 2-year period. Data regarding patient e-mail use were collected, including urgency of message, subject matter, message volume, and time of day of messaging. The duration of the pediatric rheumatologist's e-mail interactions and telephone interactions with patients was measured using a stopwatch. After 1 year of enrollment in the patient-physician e-mail service, families were mailed a 12-item satisfaction survey regarding their e-mail experience. RESULTS: A total of 306 of 328 families who were offered patient physician e-mail access enrolled, and 121 used the service. The patients sent 40% of their e-mails outside business hours. Messages that were urgent (notification of disease flare, notification of new symptoms, or parent expectation of same-day response) made up 5.7% of the e-mails sent to the physician. Messages that required emergent attention made up 0.002% of the e-mails to the physician. Answering patient questions by e-mail was 57% faster than using the telephone for the physician. The physician received 1.2 e-mails per day from patients. The families who responded to the survey agreed that patient-physician e-mail increased access to the physician and improved the quality of care. The families did not find that patient-physician e-mail distanced them from their child's doctor. CONCLUSIONS: Patient-physician e-mail is a service that patients will use given the opportunity. The e-mail service enables physicians to answer medical questions with less time spent compared with telephone messaging. In our experience in an academic pediatric subspecialty practice, patients reported enhanced communication and access with the e-mail service. PMID- 17908756 TI - Impact of clinical alerts within an electronic health record on routine childhood immunization in an urban pediatric population. AB - OBJECTIVES: The objective of this study was to test the hypothesis that clinical alerts for routine pediatric vaccinations within an electronic health record would reduce missed opportunities for vaccination and improve immunization rates for young children in an inner-city population. METHODS: A 1-year intervention study (September 1, 2004, to August 31, 2005) with historical controls was conducted in 4 urban primary care centers affiliated with an academic medical center. All children who were younger than 24 months were enrolled. Electronic health record-based clinical reminders for routine childhood vaccinations were programmed to appear prominently at every patient encounter with vaccines due. The main outcome measures were rates of captured immunization opportunities and overall immunization rates at 24 months of age. RESULTS: Immunization alerts appeared at 15,928 visits during the intervention. Alert implementation was associated with increases in captured immunization opportunities from 78.2% to 90.3% at well visits and from 11.3% to 32.0% at sick visits. Adjusted up-to-date immunization rates at 24 months of age increased from 81.7% to 90.1% from the control to intervention period. Children in the intervention group also became up to-date earlier than control patients. Patient characteristics were stable throughout the study. CONCLUSIONS: An electronic health record-based clinical alert intervention was associated with increases in captured opportunities for vaccination at both sick and well visits and significant improvements in immunization rates at 2 years of age. As electronic health records become more common in medical practice, such systems may transform immunization delivery to children. PMID- 17908757 TI - Assessing procedural skills training in pediatric residency programs. AB - OBJECTIVE: The objective of this study was to assess the opinions of pediatric program directors regarding procedural skills training of pediatric residents. METHODS: We developed a survey based on the Residency Review Committee's guidelines for procedural training. It included items about the importance of 29 procedures encountered in pediatric training, estimates of residents' competence in performing them, and the teaching of procedural skills. The survey was sent to members of the Association of Pediatric Program Directors. The primary outcome was the perceived importance for residents to achieve competence in these procedures, rated on a 10-point Likert scale. Secondary outcomes included perception of resident competence to perform procedures and educational methods used by respondents for teaching procedural skills. Associations between demographic characteristics and perceived importance or competence were also assessed. RESULTS: Surveys were sent to 139 programs, and 112 responded. Thirteen procedures were rated 8 or higher by >75% of program directors. Seven skills that were prioritized by the Residency Review Committee did not achieve this level of consensus. Respondents reported that many residents failed to achieve competence by the end of training in 9 of 13 procedures that they rated as very important, including venipuncture, neonatal intubation, and administering injections. Residents who perform the majority of venipunctures and intravenous catheter placements at their institutions were more likely to be judged competent in performing these skills than residents who do not. CONCLUSIONS: The Residency Review Committee's list of procedures does not necessarily reflect the opinions of pediatric program directors on the most essential skills for trainees. Many residents may not develop competence in several important procedures by the end of residency, most notably vascular access and life-saving skills. A more robust and standardized method is needed for teaching procedural skills and for documenting competence. PMID- 17908758 TI - Adverse birth outcome among mothers with low serum cholesterol. AB - OBJECTIVE: The objective of this study was to assess whether low maternal serum cholesterol during pregnancy is associated with preterm delivery, impaired fetal growth, or congenital anomalies in women without identified major risk factors for adverse pregnancy outcome. METHODS: Mother-infant pairs were retrospectively ascertained from among a cohort of 9938 women who were referred to South Carolina prenatal clinics for routine second-trimester serum screening. Banked sera were assayed for total cholesterol; <10th percentile of assayed values (159 mg/dL at mean gestational age of 17.6 weeks) defined a "low total cholesterol" prenatal risk category. Eligible women were aged 21 to 34 years and nonsmoking and did not have diabetes; neonates were liveborn after singleton gestations. Total cholesterol values of eligible mothers were adjusted for gestational age at screening before risk group assignment. The study population included 118 women with low total cholesterol and 940 women with higher total cholesterol. Primary analyses used multivariate regression models to compare rates of preterm delivery, fetal growth parameters, and congenital anomalies between women with low total cholesterol and control subjects with mid-total cholesterol values >10th percentile but <90th percentile. RESULTS: Prevalence of preterm delivery among mothers with low total cholesterol was 12.7%, compared with 5.0% among control subjects with mid-total cholesterol. The association of low maternal serum cholesterol with preterm birth was observed only among white mothers. Term infants of mothers with low total cholesterol weighed on average 150 g less than those who were born to control mothers. A trend of increased microcephaly risk among neonates of mothers with low total cholesterol was found. Low maternal serum cholesterol was unassociated with risk for congenital anomalies. CONCLUSIONS: Total serum cholesterol <10th population percentile was strongly associated with preterm delivery among otherwise low-risk white mothers in this pilot study population. Term infants of mothers with low total cholesterol weighed less than control infants among both racial groups. PMID- 17908759 TI - Childhood overweight increases hospital admission rates for asthma. AB - OBJECTIVES: Although childhood overweight has been associated with increased hospital lengths of stay for patients with asthma, the possible relationship between overweight and hospital admission for asthma has not been well studied. We hypothesized that overweight children who presented to the emergency department with asthma exacerbations were more likely to be admitted to the hospital than nonoverweight children. METHODS: A retrospective chart review was conducted of all children who were older than 2 years and presented to the emergency department with an asthma exacerbation in calendar year 2005. Children with chronic medical conditions other than asthma were excluded. Children were classified as nonoverweight (< or = 95% weight-for-age percentile) or overweight (> 95% weight for age). RESULTS: During the study period, there were 884 visits to the emergency department for an asthma exacerbation by 813 children; 238 (27%) were admitted to the hospital, and 33 (4%) were admitted to the ICU. Overall, hospital admission was associated with higher clinical asthma score but not with age, gender, or poverty status (as quantified as home in zip-code areas designated as "impoverished"). Overweight children (n = 202 [23%]) were significantly older (8.5 +/- 4.4 vs 7.3 +/- 4.3 years) and more likely to live in an impoverished area (37% vs 28%). Presenting clinical asthma score and therapeutic interventions in the emergency department were similar for overweight and nonoverweight children; however, overweight children were significantly more likely to be admitted to the hospital. CONCLUSIONS: Overweight children who present to the emergency department with acute asthma exacerbations are significantly more likely to be admitted to the hospital than nonoverweight children. This identifies an important area in which childhood overweight has a significant impact on the health of children with asthma. PMID- 17908760 TI - Family history of asthma and atopy: in-depth analyses of the impact on asthma and wheeze in 7- to 8-year-old children. AB - OBJECTIVES: Development of asthma in children is influenced by interactions between genetic and environmental factors. It is unclear whether paternal or maternal histories of disease confer different risks. Previous population-based studies have not stratified analyses by child gender and sensitization status. Our aim was to study in detail the hereditary component of childhood asthma. METHODS: A population-based cohort of 3430 (97% of invited) 7- to 8-year-old school children participated in an expanded International Study of Asthma and Allergy in Childhood survey, and two thirds were skin-prick tested. Heredity was defined as a family history of (1) asthma and (2) atopy (allergic rhinitis or eczema). Multivariate analyses corrected for known risk factors for asthma. RESULTS: At ages 7 to 8, prevalence of asthma was 5.3% among the children and 9.0% among the parents. In children without parental asthma or parental atopy, the prevalence of asthma was 2.8%. Corrected for parental asthma, parental atopy was a weak but significant risk factor. There were minor differences in the impact of parental disease between sensitized and nonsensitized children and between boys and girls. CONCLUSIONS: As risk factors for childhood asthma, there were major differences between parental asthma and parental atopy. Sibling asthma was only a marker of parental disease. Interactions between parental disease and the child's allergic sensitization or gender were not statistically significant. Asthma in both parents conferred a multiplicative risk, whereas the effect of parental atopy was additive, however limited. Asthma and atopy, despite their causal relationship, are separate entities and could be inherited differently. This large, population-based, and well-characterized cohort study does not confirm parent-of-origin effects found in previous studies. PMID- 17908761 TI - Do adolescent inpatient wards make a difference? Findings from a national young patient survey. AB - OBJECTIVES: There is little evidence to support the effectiveness of adolescent inpatient wards. These analyses test the hypotheses that nursing young people in adolescent wards improves aspects of quality of care and patient satisfaction compared with child or adult wards. PATIENTS AND METHODS: Secondary analyses of the national English Young Patient Survey 2004 were weighted to take account of variations in hospital size and response rate. Participants included 8855 subjects aged 12 to 17 years. Ward types (adolescent, child, and adult) were compared. Patient-reported quality-of-care indicators included rating by young people of overall care, respect, safety, confidentiality, communication, team working, noise, and leisure facilities. Logistic regression models were adjusted for gender, disability, and previous hospital admissions. RESULTS: Ten percent of 12- to 14-year-olds and 18% of 15- to 17-year-olds were nursed in an adolescent ward, 0.4% of 12- to 15-year-olds and 16% of 15- to 17-year-olds in an adult ward, with the remainder in a child ward. Compared with being in an adolescent ward, 15- to 17-year-olds were less likely to report excellent overall care in an adult ward and less likely to report feeling secure, having confidentiality maintained, feeling treated with respect, confidence in staff, appropriate information transmission, appropriate involvement in own care, and appropriate leisure facilities. Compared with being in an adolescent ward, 12- to 14-year olds were less likely to report excellent overall care in a child ward and less likely to report feeling involved in their own care. CONCLUSIONS: Dedicated adolescent inpatient wards improve aspects of quality of care for young people compared with child or adult wards, particularly for older adolescents. These data support the continued development of adolescent wards in larger general hospitals and children's hospitals. PMID- 17908762 TI - Dopamine receptor D2 gene Taq1A (C32806T) polymorphism modifies the relationship between birth weight and educational attainment in adulthood: 21-year follow-up of the Cardiovascular Risk in Young Finns study. AB - OBJECTIVE: Low birth weight is suggested to be a risk factor for a wide variety of negative outcomes, including low educational attainment, but the role of cognition-related genetic influences on this association remains unclear. The objective of this study was to study whether variation in the dopamine receptor gene (dopamine receptor D2 polymorphism, rs1800497) modifies the association between birth weight and educational attainment in adulthood. METHODS: We studied the association between birth weight (range: 1440-4980 g) and educational attainment in 659 men and 832 women aged 27 to 39. Birth weight, gestational age, and parental education were assessed at ages 6 to 18. The genotyping was performed using TaqMan 5' nuclease assay. RESULTS: After adjustment for age, parental education, and gestational age, birth weight was associated with educational attainment in men with A1/A1 or A1/A2 (n = 245) genotype but not in men carrying A2/A2 (n = 414) genotype. In women, no moderating effect of dopamine receptor D2 polymorphism was found. CONCLUSIONS: Dopamine receptor D2 genotype is suggested to modify the association between birth weight and adulthood educational attainment over the whole birth weight range so that carriers of A1 allele capitalize on optimal birth weight, whereas a low birth weight seems to be a risk among them. These data support the hypothesis that the effect of birth weight on educational attainment depends on genetic influences. Gender-related difference may refer to an environmental effect (ie, to a better goodness-of-fit between girls' school behaving and expectations of school) that may mask a genetic effect. PMID- 17908763 TI - Children's television exposure and behavioral and social outcomes at 5.5 years: does timing of exposure matter? AB - BACKGROUND: The American Academy of Pediatrics recommends children > or = 2 years of age limit daily media exposure to < or = 1 to 2 hours and not have a television set in children's bedrooms. However, there are limited prospective studies to address how timing of media exposure influences children's health. OBJECTIVE: Our goal was to examine relations among children's early, concurrent, and sustained television exposure and behavioral and social skills outcomes at 5.5 years. METHODS: We analyzed data collected prospectively from the Healthy Steps for Young Children national evaluation. Television exposure was defined as > 2 hours of daily use (at 30-33 months and 5.5 years) and television in child's bedroom (at 5.5 years). At 5.5 years, outcomes were assessed by using the Child Behavior Checklist and social skills using the Social Skills Rating System. Linear regression was used to estimate the effect of television exposure on behavioral and social skills outcomes. RESULTS: Sixteen percent of parents reported that their child watched > 2 hours of television daily at 30 to 33 months only, 15% reported > 2 hours of television daily at 5.5 years only, and 20% reported > 2 hours of television daily at both times. Forty-one percent of the children had televisions in their bedrooms at 5.5 years. In adjusted analyses, sustained television viewing was associated with behavioral outcomes. Concurrent television exposure was associated with fewer social skills. For children with heavy television viewing only in early childhood, there was no consistent relation with behavioral or social skills outcomes. Having a television in the bedroom was associated with sleep problems and less emotional reactivity at 5.5 years but was not associated with social skills. CONCLUSIONS: Sustained exposure is a risk factor for behavioral problems, whereas early exposure that is subsequently reduced presents no additional risk. For social skills, concurrent exposure was more important than sustained or early exposure. Considering the timing of media exposure is vital for understanding the consequences of early experiences and informing prevention strategies. PMID- 17908764 TI - Sensitivity of amplitude-integrated electroencephalography for neonatal seizure detection. AB - BACKGROUND: Conventional electroencephalography remains the gold standard for the diagnosis and quantification of neonatal seizures. However, amplitude-integrated electroencephalography (aEEG) is being introduced to neonatal intensive care as an adjunct for neonatal seizure detection. OBJECTIVES: This study's purpose was to determine the sensitivity of neonatal seizure detection in a single electroencephalogram channel (C3-->C4), used to simulate the raw signal from which aEEG is derived. We also aimed to determine the sensitivity of seizure detection by neonatologists by using aEEG and to establish those neonatal seizure characteristics that are associated with their correct detection by aEEG. METHODS: Conventional electroencephalograms with neonatal seizures were reviewed for electroencephalogram background and neonatal seizure characteristics (site of onset, duration, and peak-to-peak amplitude). The presence, duration, and peak-to peak amplitude of each seizure were simultaneously noted in a single electroencephalogram channel (C3-->C4). aEEGs generated from this channel were reviewed for background and seizures by 6 neonatologists with varying aEEG interpretation expertise. RESULTS: A total of 851 neonatal seizures from 125 conventional electroencephalograms were analyzed. The patients' conceptional ages were 34 to 50 weeks. Because 94% of the conventional electroencephalograms had > or = 1 neonatal seizure visible in C3-->C4, and 78% of all neonatal seizures appeared in the C3-->C4 channel, the theoretical sensitivity of seizure detection in a single electroencephalogram channel was high. However, seizures were briefer and lower in amplitude in C3-->C4 compared with conventional electroencephalography. Neonatologists identified seizures in 22% to 57% of the 125 records of neonatal seizure. They detected 12% to 38% of the 851 individual seizures. Multivariate analysis revealed that the appearance of seizures in C3- >C4, neonatal seizure duration, seizure amplitude, seizure count per hour, and neonatologists' experience with aEEG interpretation all correlated with neonatal seizure detection. CONCLUSIONS: Even among physicians who have extensive experience, many neonatal seizures are difficult to detect on an aEEG, especially when they are infrequent, brief, or of low amplitude. PMID- 17908765 TI - Very low birth weight increases risk for sleep-disordered breathing in young adulthood: the Helsinki Study of Very Low Birth Weight Adults. AB - OBJECTIVE: We investigated whether very low birth weight (<1500 g) is associated with the risk of sleep-disordered breathing in young adulthood. METHODS: The study was a retrospective longitudinal study of 158 young adults born with very low birth weight and 169 term-born control subjects (aged 18.5-27.1 years). The principal outcome variable was sleep-disordered breathing defined as chronic snoring. RESULTS: The crude prevalence of chronic snoring was similar in both groups: 15.8% for the very low birth weight group versus 13.6% for the control group. However, after controlling for the confounding variables in multivariate logistic regression models (age, gender, current smoking, parental education, height, BMI, and depression), chronic snoring was 2.2 times more likely in the very low birth weight group compared with the control group. In addition, maternal smoking during pregnancy was significantly and independently of very low birth weight related to risk of sleep-disordered breathing. Maternal preeclampsia, standardized birth weight, and, for very low birth weight infants, small-for-gestational-age status were not related to sleep-disordered breathing. CONCLUSIONS: Premature infants with very low birth weight have a twofold risk of sleep-disordered breathing as young adults. In addition, maternal smoking during pregnancy increases the risk of sleep-disordered breathing by more than twofold. PMID- 17908766 TI - Neurodevelopmental outcome in survivors of periventricular hemorrhagic infarction. AB - OBJECTIVES: Periventricular hemorrhagic infarction is a serious complication of germinal matrix-intraventricular hemorrhage in premature infants. Our objective was to determine the neurodevelopmental and adaptive outcomes of periventricular hemorrhagic infarction survivors and identify early cranial ultrasound predictors of adverse outcome. METHODS: We retrospectively evaluated all cranial ultrasounds of 30 premature infants with periventricular hemorrhagic infarction and assigned a cranial ultrasound-based periventricular hemorrhagic infarction severity score (range: 0-3) on the basis of whether periventricular hemorrhagic infarction (1) involved > or = 2 territories, (2) was bilateral, or (3) caused midline shift. We then performed neuromotor, visual function, and developmental evaluations (Mullen Scales of Early Learning, Vineland Adaptive Behavior Scale). Developmental scores below 2 SD from the mean were defined as abnormal. RESULTS: Median adjusted age at evaluation was 30 months (range: 12-66 months). Eighteen subjects (60%) had abnormal muscle tone, and 7 (26%) had visual field defects. Developmental delays involved gross motor (22 [73%]), fine motor (17 [59%]), visual receptive (13 [46%]), expressive language (11 [38%]), and cognitive (14 [50%]) domains. Impairment in daily living and socialization was documented in 10 (33%) and 6 (20%) infants, respectively. Higher cranial ultrasound-based periventricular hemorrhagic infarction severity scores predicted microcephaly and abnormalities in gross motor, visual receptive, and cognitive function. CONCLUSIONS: In the current era, two thirds of periventricular hemorrhagic infarction survivors develop significant cognitive and/or motor abnormalities, whereas adaptive skills are relatively spared. Higher cranial ultrasound-based periventricular hemorrhagic infarction severity scores predict worse outcome in several modalities and may prove to be a valuable tool for prognostication. PMID- 17908767 TI - Motor and executive function at 6 years of age after extremely preterm birth. AB - BACKGROUND: Studies of very preterm infants have demonstrated impairments in multiple neurocognitive domains. We hypothesized that neuromotor and executive function deficits may independently contribute to school failure. METHODS: We studied children who were born at < or = 25 completed weeks' gestation in the United Kingdom and Ireland in 1995 at early school age. Children underwent standardized cognitive and neuromotor assessments, including the Kaufman Assessment Battery for Children and NEPSY, and a teacher-based assessment of academic achievement. RESULTS: Of 308 surviving children, 241 (78%) were assessed at a median age of 6 years 4 months. Compared with 160 term classmates, 180 extremely preterm children without cerebral palsy and attending mainstream school performed less well on 3 simple motor tasks: posting coins, heel walking, and 1 leg standing. They more frequently had non-right-hand preferences (28% vs 10%) and more associated/overflow movements during motor tasks. Standardized scores for visuospatial and sensorimotor function performance differed from classmates by 1.6 and 1.1 SDs of the classmates' scores, respectively. These differences attenuated but remained significant after controlling for overall cognitive scores. Cognitive, visuospatial scores, and motor scores explained 54% of the variance in teachers' ratings of performance in the whole set; in the extremely preterm group, additional variance was explained by attention-executive tasks and gender. CONCLUSIONS: Impairment of motor, visuospatial, and sensorimotor function, including planning, self-regulation, inhibition, and motor persistence, contributes excess morbidity over cognitive impairment in extremely preterm children and contributes independently to poor classroom performance at 6 years of age. PMID- 17908768 TI - Overweight, race, and psychological distress in children in the Childhood Asthma Management Program. AB - OBJECTIVE: The purpose of this work was to determine whether overweight in youth with mild-to-moderate asthma occurs with increased frequency and is accompanied by impaired psychological functioning. PATIENTS AND METHODS: The interrelationships among BMI and demographic and psychological characteristics were examined in 1005 children (aged 5-12 years) enrolled in the Childhood Asthma Management Program and seen for repeated visits over 4 1/2 years. RESULTS: Baseline rates of overweight (BMI for age: > or = 95th percentile) were comparable, but rates of overweight risk (BMI for age: 85th to < 95th percentile) among children in the Childhood Asthma Management Program were elevated in comparison with the general population of children in the United States. Rates of overweight and overweight risk did not increase over the course of the longitudinal study. Overweight and overweight risk were more frequent among black and Hispanic than white children, although they were not higher relative to same race groups in the general population. Overweight at baseline was associated with lower IQ, more social withdrawal, and greater internalized psychological distress. As the children became older, the overweight group demonstrated increased evidence of behavior problems and decreased physical activity. CONCLUSIONS: This study identifies an increase in overweight risk but not overweight in children with mild-to-moderate asthma. Comorbidity between asthma and overweight may be underestimated, because children with severe asthma and those from impoverished backgrounds were not represented in this sample. For the 14% of children who were overweight, some associated psychological difficulties were present in childhood, and additional problems were seen during adolescence. These results suggest a need for programs that encourage greater vigilance and intervention for overweight children with asthma. PMID- 17908769 TI - Genetic polymorphisms in immunoresponse genes TNFA, IL6, IL10, and TLR4 are associated with recurrent acute otitis media. AB - OBJECTIVE: Cytokines and other inflammatory mediators are involved in the pathogenesis of otitis media. We hypothesized that polymorphisms in inflammatory response genes contribute to the increased susceptibility to acute otitis media in otitis-prone children. PATIENTS AND METHODS: DNA samples from 348 children with > or = 2 acute otitis media episodes, who were participating in a randomized, controlled vaccination trial, and 463 healthy adult controls were included. Polymorphisms in TNFA, IL1B, IL4, IL6, IL10, IL8, NOS2A, C1INH, PARP, TLR2, and TLR4 were genotyped. Genotype distributions in children with recurrent acute otitis media were compared with those in controls. Within the patient group, the number of acute otitis media episodes before vaccination and the clinical and immunologic response to pneumococcal conjugate vaccinations were analyzed. RESULTS: The IL6-174 G/G genotype was overrepresented in children with acute otitis media when compared with controls. In the patient group, TNFA promoter genotypes -238 G/G and -376 G/G and the TLR4 299 A/A genotype were associated with an otitis-prone condition. Furthermore, lower specific anticapsular antibody production after complete vaccination was observed in patients with the TNFA-238 G/G genotype or TNFA-863 A allele carriage. Finally, the IL10-1082 A/A genotype contributed to protection from the recurrence of acute otitis media after pneumococcal vaccination. CONCLUSIONS: Variation in innate immunoresponse genes such as TNFA-863A, TNFA-376G, TNFA-238G, IL10-1082 A, and IL6-174G alleles in the promoter sequences may result in altered cytokine production that leads to altered inflammatory responses and, hence, contributes to an otitis-prone condition. PMID- 17908770 TI - Disease progression in Hutchinson-Gilford progeria syndrome: impact on growth and development. AB - OBJECTIVES: Hutchinson-Gilford progeria syndrome is a rare and uniformly fatal segmental "premature aging" disease that affects a variety of organ systems. We sought to more clearly define the bone and weight abnormalities in patients with progeria as potential outcome parameters for prospective clinical trials. PATIENTS AND METHODS: We collected and analyzed longitudinal medical information, both retrospectively and prospectively, from a total of 41 children with Hutchinson-Gilford progeria syndrome spanning 14 countries, from the Progeria Research Foundation Medical and Research Database at the Brown University Center for Gerontology. RESULTS: In addition to a number of previously well-defined phenotypic findings in children with progeria, this study identified abnormalities in the eruption of secondary incisors lingually and palatally in the mandible and maxilla, respectively. Although bony structures appeared normal in early infancy, clavicular resorption, coxa valga, avascular necrosis of the femoral head, modeling abnormalities of long bones with slender diaphyses, flared metaphyses, and overgrown epiphyses developed. Long bones showed normal cortical thickness centrally and progressive focal demineralization peripherally. The most striking finding identified in the retrospective data set of 35 children was an average weight increase of only 0.44 kg/year, beginning at approximately 24 months of age and persisting through life, with remarkable intrapatient linearity. This rate is >2 SD below normal weight gain for any corresponding age and sharply contrasts with the parabolic growth pattern for normal age- and gender-matched children. This finding was also confirmed prospectively. CONCLUSIONS: Our analysis shows evidence of a newly identified abnormal growth pattern for children with Hutchinson-Gilford progeria syndrome. The skeletal and dental findings are suggestive of a developmental dysplasia rather than a classical aging process. The presence of decreased and linear weight gain, maintained in all of the patients after the age of 2 years, provides the ideal parameter on which altered disease status can be assessed in clinical trials. PMID- 17908771 TI - New approaches to progeria. AB - Progeria (Hutchinson-Gilford progeria syndrome) is a rare genetic disorder that offers considerable insight into the biology of premature aging. This review summarizes the clinical characteristics of this disease and the underlying mutation in the lamin A (LMNA) gene that results in this phenotype. Modifications in the processing of prelamin A through alterations in farnesylation are detailed, because this pathway offers a possible drug target. Finally, discussion of an ongoing clinical trial for these children, including possible parameters for evaluation, are discussed. In the span of less than a decade, this disease has progressed from an interesting phenotype to one in which the gene defect has been identified, animal models have been created and tested with drugs that target the primary disease pathway, and significant clinical baseline data for the support of a clinical trial have been obtained. PMID- 17908772 TI - Parent presence during complex invasive procedures and cardiopulmonary resuscitation: a systematic review of the literature. AB - We conducted a systematic review of the literature on parent presence during complex invasive pediatric procedures and/or resuscitation. We identified 15 studies that met our inclusion criteria. All studies were summarized chronologically according to level of evidence. The studies all demonstrated that parents prefer to have the choice about whether they remain at their child's side during complex invasive procedures and resuscitation, but they also revealed that apprehensions and controversy abound among clinicians regarding this practice. Despite the endorsements of the American Academy of Pediatrics and the Society of Critical Care Medicine and the recommendations of the American Heart Association, few pediatric institutions have drafted guidelines, conducted clinical education, or committed sufficient staff resources to fully support this practice. We present this review not only to illustrate the various perspectives of parents/guardians, clinicians, and pediatric patients themselves that have been reported to date but also to encourage more research so that the practice can be performed safely and benefit parents, their children, and clinicians alike. PMID- 17908773 TI - Pseudo-asthma: when cough, wheezing, and dyspnea are not asthma. AB - Although asthma is the most common cause of cough, wheeze, and dyspnea in children and adults, asthma is often attributed inappropriately to symptoms from other causes. Cough that is misdiagnosed as asthma can occur with pertussis, cystic fibrosis, primary ciliary dyskinesia, airway abnormalities such as tracheomalacia and bronchomalacia, chronic purulent or suppurative bronchitis in young children, and habit-cough syndrome. The respiratory sounds that occur with the upper airway obstruction caused by the various manifestations of the vocal cord dysfunction syndrome or the less common exercise-induced laryngomalacia are often mischaracterized as wheezing and attributed to asthma. The perception of dyspnea is a prominent symptom of hyperventilation attacks. This can occur in those with or without asthma, and patients with asthma may not readily distinguish the perceived dyspnea of a hyperventilation attack from the acute airway obstruction of asthma. Dyspnea on exertion, in the absence of other symptoms of asthma or an unequivocal response to albuterol, is most likely a result of other causes. Most common is the dyspnea associated with normal exercise limitation, but causes of dyspnea on exertion can include other physiologic abnormalities including exercise-induced vocal cord dysfunction, exercise-induced laryngomalacia, exercise-induced hyperventilation, and exercise induced supraventricular tachycardia. A careful history, attention to the nature of the respiratory sounds that are present, spirometry, exercise testing, and blood-gas measurement provide useful data to sort out the various causes and avoid inappropriate treatment of these pseudo-asthma clinical manifestations. PMID- 17908775 TI - Viral testing and isolation of patients with bronchiolitis. PMID- 17908774 TI - Health care epidemiology perspective on the October 2006 recommendations of the Subcommittee on Diagnosis and Management of Bronchiolitis. PMID- 17908776 TI - Professionalism in pediatrics: statement of principles. AB - The purpose of this statement is to delineate the concept of professionalism within the context of pediatrics and to provide a brief statement of principles to guide the behavior and professional practice of pediatricians. PMID- 17908777 TI - Year 2007 position statement: Principles and guidelines for early hearing detection and intervention programs. PMID- 17908778 TI - Nondiscrimination in pediatric health care. AB - This policy statement is a revision of a 2001 statement and articulates the positions of the American Academy of Pediatrics on nondiscrimination in pediatric health care. It addresses both pediatricians who provide health care and the infants, children, adolescents, and young adults whom they serve. PMID- 17908779 TI - Unsuspected consequences of the adolescent overweight epidemic. PMID- 17908780 TI - Refrigerator-stable and frozen formulation of ProQuad. PMID- 17908781 TI - Criticism of infant swimming practice is political, not scientific. PMID- 17908782 TI - Pediatricians and counseling about the military. PMID- 17908783 TI - Hypnosis in children. PMID- 17908784 TI - Pediatricians and counseling about the military. PMID- 17908785 TI - Pediatricians and counseling about the military. PMID- 17908786 TI - Cardiovascular effects of sibutramine. PMID- 17908787 TI - Propofol compared with the morphine, atropine, and suxamethonium regimen as induction agents for neonatal endotracheal intubation: a randomized, controlled trial. PMID- 17908788 TI - Malpractice claims involving pediatricians. PMID- 17908789 TI - Functional analysis of KSRP interaction with the AU-rich element of interleukin-8 and identification of inflammatory mRNA targets. AB - mRNA stability is a major determinant of inflammatory gene expression. Rapid degradation of interleukin-8 (IL-8) mRNA is imposed by a bipartite AU-rich element (ARE) in the 3' untranslated region (R. Winzen et al., Mol. Cell. Biol. 24:4835-4847, 2004). Small interfering RNA-mediated knockdown of the ARE-binding protein KSRP resulted in stabilization of IL-8 mRNA or of a beta-globin reporter mRNA containing the IL-8 ARE. Rapid deadenylation was impaired, indicating a crucial role for KSRP in this step of mRNA degradation. The two IL-8 ARE domains both contribute to interaction with KSRP, corresponding to the importance of both domains for rapid degradation. Exposure to the inflammatory cytokine IL-1 has been shown to stabilize IL-8 mRNA through p38 mitogen-activated protein (MAP) kinase and MK2. IL-1 treatment impaired the interaction of KSRP with the IL-8 ARE in a manner dependent on p38 MAP kinase but apparently independent of MK2. Instead, evidence that TTP, a target of MK2, can also destabilize the IL-8 ARE reporter mRNA is presented. In a comprehensive approach to identify mRNAs controlled by KSRP, two criteria were evaluated by microarray analysis of (i) association of mRNAs with KSRP in pulldown assays and (ii) increased amounts in KSRP knockdown cells. According to both criteria, a group of 100 mRNAs is controlled by KSRP, many of which are unstable and encode proteins involved in inflammation. These results indicate that KSRP functions as a limiting factor in inflammatory gene expression. PMID- 17908790 TI - Ubiquitination and degradation of mutant p53. AB - While wild-type p53 is normally a rapidly degraded protein, mutant forms of p53 are stabilized and accumulate to high levels in tumor cells. In this study, we show that mutant and wild-type p53 proteins are ubiquitinated and degraded through overlapping but distinct pathways. While Mdm2 can drive the degradation of both mutant and wild-type p53, our data suggest that the ability of Mdm2 to function as a ubiquitin ligase is less important in the degradation of mutant p53, which is heavily ubiquitinated in an Mdm2-independent manner. Our initial attempts to identify ubiquitin ligases that are responsible for the ubiquitination of mutant p53 have suggested a role for the chaperone-associated ubiquitin ligase CHIP (C terminus of Hsc70-interacting protein), although other unidentified ubiquitin ligases also appear to contribute. The contribution of Mdm2 to the degradation of mutant p53 may reflect the ability of Mdm2 to deliver the ubiquitinated mutant p53 to the proteasome. PMID- 17908791 TI - Regulation of the Il4 gene is independently controlled by proximal and distal 3' enhancers in mast cells and basophils. AB - Mast cells and basophils are known to be a critical interleukin 4 (IL-4) source for establishing Th2 protective responses to parasitic infections. Chromatin structure and histone modification patterns in the Il13/Il4 locus of mast cells were similar to those of IL-4-producing type 2 helper T cells. However, using a transgenic approach, we found that Il4 gene expression was distinctly regulated by individual cis regulatory elements in cell types of different lineages. The distal 3' element contained conserved noncoding sequence 2 (CNS-2), which was a common enhancer for memory phenotype T cells, NKT cells, mast cells, and basophils. Targeted deletion of CNS-2 compromised production of IL-4 and several Th2 cytokines in connective-tissue-type and immature-type mast cells but not in basophils. Interestingly, the proximal 3' element containing DNase I hypersensitive site 4 (HS4), which controls Il4 gene silencing in T-lineage cells, exhibited selective enhancer activity in basophils. These results indicate that CNS-2 is an essential enhancer for Il4 gene transcription in mast cell but not in basophils. The transcription of the Il4 gene in mast cells and basophils is independently regulated by CNS-2 and HS4 elements that may be critical for lineage-specific Il4 gene regulation in these cell types. PMID- 17908792 TI - Domain architecture of the catalytic subunit in the ISW2-nucleosome complex. AB - ATP-dependent chromatin remodeling has an important role in the regulation of cellular differentiation and development. For the first time, a topological view of one of these complexes has been revealed, by mapping the interactions of the catalytic subunit Isw2 with nucleosomal and extranucleosomal DNA in the complex with all four subunits of ISW2 bound to nucleosomes. Different domains of Isw2 were shown to interact with the nucleosome near the dyad axis, another near the entry site of the nucleosome, and another with extranucleosomal DNA. The conserved DEXD or ATPase domain was found to contact the superhelical location 2 (SHL2) of the nucleosome, providing a direct physical connection of ATP hydrolysis with this region of nucleosomes. The C terminus of Isw2, comprising the SLIDE (SANT-like domain) and HAND domains, was found to be associated with extranucleosomal DNA and the entry site of nucleosomes. It is thus proposed that the C-terminal domains of Isw2 are involved in anchoring the complex to nucleosomes through their interactions with linker DNA and that they facilitate the movement of DNA along the surface of nucleosomes. PMID- 17908793 TI - Functional roles of Otx2 transcription factor in postnatal mouse retinal development. AB - We previously reported that Otx2 is essential for photoreceptor cell fate determination; however, the functional role of Otx2 in postnatal retinal development is still unclear although it has been reported to be expressed in retinal bipolar cells and photoreceptors at postnatal stages. In this study, we first examined the roles of Otx2 in the terminal differentiation of photoreceptors by analyzing Otx2; Crx double-knockout mice. In Otx2+/-; Crx-/- retinas, photoreceptor degeneration and downregulation of photoreceptor-specific genes were much more prominent than in Crx-/- retinas, suggesting that Otx2 has a role in the terminal differentiation of the photoreceptors. Moreover, bipolar cells decreased in the Otx2+/-; Crx-/- retina, suggesting that Otx2 is also involved in retinal bipolar-cell development. To further investigate the role of Otx2 in bipolar-cell development, we generated a postnatal bipolar-cell-specific Otx2 conditional-knockout mouse line. Immunohistochemical analysis of this line showed that the expression of protein kinase C, a marker of mature bipolar cells, was significantly downregulated in the retina. Electroretinograms revealed that the electrophysiological function of retinal bipolar cells was impaired as a result of Otx2 ablation. These data suggest that Otx2 plays a functional role in the maturation of retinal photoreceptor and bipolar cells. PMID- 17908794 TI - Compromised intestinal lipid absorption in mice with a liver-specific deficiency of liver receptor homolog 1. AB - Bile acids (BAs) are water-soluble end products from cholesterol metabolism and are essential for efficient absorption of dietary lipids. By using targeted somatic mutagenesis of the nuclear receptor liver receptor homolog 1 (LRH-1) in mouse hepatocytes, we demonstrate here that LRH-1 critically regulates the physicochemical properties of BAs. The absence of LRH-1 and subsequent deficiency of Cyp8b1 eliminate the production of cholic acid and its amino acid conjugate taurocholic acid and increase the relative amounts of less amphipathic BA species. Intriguingly, while the expression of Cyp8b1 is almost extinguished in the livers of mice that lack LRH-1, the expression of the rate-limiting enzyme of BA synthesis, i.e., Cyp7a1, remains unchanged. The profound remodeling of the BA composition significantly reduces the efficacy of intestinal absorption of lipids and reuptake of BAs and facilitates the removal of lipids from the body. Our studies unequivocally demonstrate a pivotal role for LRH-1 in determining the composition of BAs, which, in turn has major consequences on whole-body lipid homeostasis. PMID- 17908795 TI - Inhibition of interleukin-1beta-induced group IIA secretory phospholipase A2 expression by peroxisome proliferator-activated receptors (PPARs) in rat vascular smooth muscle cells: cooperation between PPARbeta and the proto-oncogene BCL-6. AB - The inflammation that occurs during atherosclerosis is characterized by the release of large amounts of group IIA secretory phospholipase A2 (sPLA2-IIA). This study was designed to define the function of the three peroxisome proliferator-activated receptors (PPARs) on sPLA2 expression in vascular smooth muscle cells (VSMCs). We found that PPAR ligands decreased sPLA2-IIA activity and inhibited mRNA accumulation under inflammatory conditions. Furthermore, interleukin-1beta-induced sPLA2-IIA promoter activity was inhibited by the three PPAR ligands and in a similar way when cells were cotransfected with PPARalpha, PPARbeta, or PPARgamma, plus retinoid X receptor alpha (RXRalpha). Our study revealed that the regulation of sPLA2-IIA gene transcription by PPARalpha/RXR and PPARgamma/RXR heterodimers requires an interaction with a PPAR response element (PPRE) of the sPLA2-IIA promoter. In contrast, PPARbeta operates through a PPRE independent mechanism. In addition, we demonstrated that VSMCs expressed the transcriptional repressor BCL-6. Overexpression of BCL-6 markedly reduced sPLA2 IIA promoter activity in VSMCs, while a dominant negative form of BCL-6 abrogated sPLA2 repression by PPARbeta. The PPARbeta agonist induced a BCL-6 binding to the sPLA2 promoter in VSMCs under inflammatory conditions. The knockdown of BCL-6 by short interfering RNA abolished the inhibitory effect of the PPARbeta ligand on sPLA2 activity and prostaglandin E2 release. Thus, the inhibition of sPLA2-IIA activity by PPARbeta agonists may provide a promising approach to impacting the initiation and progression of atherosclerosis. PMID- 17908796 TI - Differential modification of p27Kip1 controls its cyclin D-cdk4 inhibitory activity. AB - Whether p27 is a cyclin D-cdk4/6 inhibitor or not is controversial, and how it might switch between these two modes is unknown. Arguing for a two-state mechanism, we show that p27 bound to cyclin D-cdk4 can be both inhibitory and noninhibitory, due to its differential-growth-state-dependent tyrosine phosphorylation. We found that p27 from proliferating cells was noninhibitory but that p27 from arrested cells was inhibitory, and the transition from a bound noninhibitor to a bound inhibitor was not due to an increase in p27 concentration. Rather, two tyrosine residues (Y88 and Y89) in p27's cdk interaction domain were phosphorylated preferentially in proliferating cells, which converted p27 to a noninhibitor. Concordantly, mutation of these sites rendered p27 resistant to phosphorylation and locked it into the bound-inhibitor mode in vivo and in vitro. Y88 was directly phosphorylated in vitro by the tyrosine kinase Abl, which converted p27 to a cdk4-bound noninhibitor. These data show that the growth-state-dependent tyrosine phosphorylation of p27 modulates its inhibitory activity in vivo. PMID- 17908798 TI - Activation of the G2/M-specific gene CLB2 requires multiple cell cycle signals. AB - In budding yeast (Saccharomyces cerevisiae), the periodic expression of the G2/M specific gene CLB2 depends on a DNA binding complex that mediates its repression during G1 and activation from the S phase to the exit of mitosis. The switch from low to high expression levels depends on the transcriptional activator Ndd1. We show that the inactivation of the Sin3 histone deacetylase complex bypasses the essential role of Ndd1 in cell cycle progression. Sin3 and its catalytic subunit Rpd3 associate with the CLB2 promoter during the G1 phase of the cell cycle. Both proteins dissociate from the promoter at the onset of the S phase and reassociate during G2 phase. Sin3 removal coincides with a transient increase in histone H4 acetylation followed by the expulsion of at least one nucleosome from the promoter region. Whereas the first step depends on Cdc28/Cln1 activity, Ndd1 function is required for the second step. Since the removal of Sin3 is independent of Ndd1 recruitment and Cdc28/Clb activity it represents a unique regulatory step which is distinct from transcriptional activation. PMID- 17908797 TI - Tissue- and nuclear receptor-specific function of the C-terminal LXXLL motif of coactivator NCoA6/AIB3 in mice. AB - Although the LXXLL motif of nuclear receptor (NR) coactivators is essential for interaction with NRs, its role has not been assessed in unbiased animal models. The nuclear receptor coactivator 6 (NCoA6; also AIB3, PRIP, ASC-2, TRBP, RAP250, or NRC) is a coactivator containing an N-terminal LXXLL-1 (L1) and a C-terminal L2. L1 interacts with many NRs, while L2 interacts with the liver X receptor alpha (LXRalpha) and the estrogen receptor alpha (ERalpha). We generated mice in which L2 was mutated into AXXAL (L2m) to disrupt its interaction with LXRalpha and ERalpha. NCoA6(L2m/L2m) mice exhibited normal reproduction, mammary gland morphogenesis, and ERalpha target gene expression. In contrast, when treated with an LXRalpha agonist, lipogenesis and the LXRalpha target gene expression were significantly reduced in NCoA6(L2m/L2m) mice. The induction of Cyp7A1 expression by a high-cholesterol diet was impaired in NCoA6(L2m/L2m) mice, which reduced bile acid synthesis in the liver and excretion in the feces and resulted in cholesterol accumulation in the liver and blood. These results demonstrate that L2 plays a tissue- and NR-specific role: it is required for NCoA6 to mediate LXRalpha-regulated lipogenesis and cholesterol/bile acid homeostasis in the liver but not required for ERalpha function in the mammary gland. PMID- 17908799 TI - RACK1 targets the extracellular signal-regulated kinase/mitogen-activated protein kinase pathway to link integrin engagement with focal adhesion disassembly and cell motility. AB - The extracellular signal-regulated kinase (ERK) cascade is activated in response to a multitude of extracellular signals and converts these signals into a variety of specific biological responses, including cell differentiation, cell movement, cell division, and apoptosis. The specificity of the biological response is likely to be controlled in large measure by the localization of signaling, thus enabling ERK activity to be directed towards specific targets. Here we show that the RACK1 scaffold protein functions specifically in integrin-mediated activation of the mitogen-activated protein kinase/ERK cascade and targets active ERK to focal adhesions. We found that RACK1 associated with the core kinases of the ERK pathway, Raf, MEK, and ERK, and that attenuation of RACK1 expression resulted in a decrease in ERK activity in response to adhesion but not in response to growth factors. RACK1 silencing also caused a reduction of active ERK in focal adhesions, an increase in focal adhesion length, a decreased rate of focal adhesion disassembly, and decreased motility. Our data further suggest that focal adhesion kinase is an upstream activator of the RACK1/ERK pathway. We suggest that RACK1 tethers the ERK pathway core kinases and channels signals from upstream activation by integrins to downstream targets at focal adhesions. PMID- 17908800 TI - Identification of candidate angiogenic inhibitors processed by matrix metalloproteinase 2 (MMP-2) in cell-based proteomic screens: disruption of vascular endothelial growth factor (VEGF)/heparin affin regulatory peptide (pleiotrophin) and VEGF/Connective tissue growth factor angiogenic inhibitory complexes by MMP-2 proteolysis. AB - Matrix metalloproteinases (MMPs) exert both pro- and antiangiogenic functions by the release of cytokines or proteolytically generated angiogenic inhibitors from extracellular matrix and basement membrane remodeling. In the Mmp2-/- mouse neovascularization is greatly reduced, but the mechanistic aspects of this remain unclear. Using isotope-coded affinity tag labeling of proteins analyzed by multidimensional liquid chromatography and tandem mass spectrometry we explored proteome differences between Mmp2-/- cells and those rescued by MMP-2 transfection. Proteome signatures that are hallmarks of proteolysis revealed cleavage of many known MMP-2 substrates in the cellular context. Proteomic evidence of MMP-2 processing of novel substrates was found. Insulin-like growth factor binding protein 6, follistatin-like 1, and cystatin C protein cleavage by MMP-2 was biochemically confirmed, and the cleavage sites in heparin affin regulatory peptide (HARP; pleiotrophin) and connective tissue growth factor (CTGF) were sequenced by matrix-assisted laser desorption ionization-time of flight mass spectrometry. MMP-2 processing of HARP and CTGF released vascular endothelial growth factor (VEGF) from angiogenic inhibitory complexes. The cleaved HARP N-terminal domain increased HARP-induced cell proliferation, whereas the HARP C-terminal domain was antagonistic and decreased cell proliferation and migration. Hence the unmasking of cytokines, such as VEGF, by metalloproteinase processing of their binding proteins is a new mechanism in the control of cytokine activation and angiogenesis. PMID- 17908801 TI - Light and electron microscopy characteristics of the muscle of patients with SURF1 gene mutations associated with Leigh disease. AB - AIMS: Leigh syndrome (LS) is characterised by almost identical brain changes despite considerable causal heterogeneity. SURF1 gene mutations are among the most frequent causes of LS. Although deficiency of cytochrome c oxidase (COX) is a typical feature of the muscle in SURF1-deficient LS, other abnormalities have been rarely described. The aim of the present work is to assess the skeletal muscle morphology coexisting with SURF1 mutations from our own research and in the literature. METHODS: Muscle samples from 21 patients who fulfilled the criteria of LS and SURF1 mutations (14 homozygotes and 7 heterozygotes of c.841delCT) were examined by light and electron microscopy. RESULTS: Diffuse decreased activity or total deficit of COX was revealed histochemically in all examined muscles. No ragged red fibres (RRFs) were seen. Lipid accumulation and fibre size variability were found in 14 and 9 specimens, respectively. Ultrastructural assessment showed several mitochondrial abnormalities, lipid deposits, myofibrillar disorganisation and other minor changes. In five cases no ultrastructural changes were found. Apart from slight correlation between lipid accumulation shown by histochemical and ultrastructural techniques, no other correlations were revealed between parameters investigated, especially between severity of morphological changes and the patient's age at the biopsy. CONCLUSION: Histological and histochemical features of muscle of genetically homogenous SURF1-deficient LS were reproducible in detection of COX deficit. Minor muscle changes were not commonly present. Also, ultrastructural abnormalities were not a consistent feature. It should be emphasised that SURF1 deficient muscle assessed in the light and electron microscopy panel may be interpreted as normal if COX staining is not employed. PMID- 17908802 TI - A detailed morphologic and immunohistochemical comparison of pre- and postmenopausal endometriosis. AB - BACKGROUND: Endometriosis depends on oestrogenic stimulation for its continued growth, accounting for its prevalence during reproductive years. There is doubt among clinicians regarding its existence in postmenopausal women in the absence of exogenous or endogenous sex hormones. We postulated that endometriosis occurring in postmenopausal women would show a different morphologic and immunohistochemical profile to the disease occurring in premenopausal women. METHODS: We reviewed the most recent 100 cases of endometriosis in our department plus all cases occurring in women aged > or =50 dating back to 1999. Cases were divided into <50 (n = 91), 50-59 (8) and > or =60 (6), and analysed for extent of disease, proportions of epithelium and stroma, and amount of haemorrhage. Immunohistochemistry for oestrogen receptor (OR), progesterone receptor (PR) and CD10 was performed and analysed on all cases aged > or =50 and on controls <50. RESULTS: There was statistically less disease in older women when cases occurring in the cervix and in scars were excluded (p = 0.0191). There was no statistical difference in the proportions of epithelium or stroma, but there was a statistical difference in the amount of haemorrhage (p = 0.0154) with older women showing less haemorrhage. There was no significant difference in immunohistochemical profile. CONCLUSIONS: Endometriosis does occur in postmenopausal women but is less common, is present in smaller volumes, and is less active. It has the same immunohistochemical profile as the disease occurring in premenopausal women and we infer from this that it has the potential to reactivate given the appropriate stimulation. PMID- 17908804 TI - Detection of epidermal growth factor receptor gene mutations in cytology specimens from patients with non-small cell lung cancer utilising high-resolution melting amplicon analysis. AB - BACKGROUND: Activating epidermal growth factor receptor (EGFR) mutations have been implicated in non-small cell lung cancer (NSCLC), and have also been clinically correlated with patient sensitivity to targeted EGFR inhibitors. AIM: To describe a technique for determining EGFR mutation status on archival fine needle aspirate (FNA) specimens from advanced NSCLC patients. METHODS: Eleven archival FNA slides from patients with advanced NSCLC were examined for diagnostic material to identify tumour cell-enriched regions. EGFR mutation status was determined using a slide-scrape DNA extraction protocol of selected tumour cell regions on the smear slides, followed by real time PCR and high resolution melt analysis (HRMAA) of EGFR exons 18, 19, 20, and 21, followed by sequence analysis. RESULTS: All DNA samples were successfully amplified by PCR. Three adenocarcinoma patient samples contained EGFR mutations in exon 19 (L747 P753insS). One of the three had an additional exon 19 mutation (A755D). CONCLUSIONS: Archival cytology slides from patients with NSCLC can be used to determine EGFR mutation status by PCR, HRMAA, and sequencing. The ability to use archival cytology slides greatly increases the potential material available for molecular analysis in diagnosis and selection of patients for targeted therapeutic agents. PMID- 17908803 TI - E2F-1 overexpression correlates with decreased proliferation and better prognosis in adenocarcinomas of Barrett oesophagus. AB - BACKGROUND: E2F-1 expression is positively associated with tumour growth in oesophageal squamous-cell carcinomas (OSCC), while it exhibits oncosuppressive features in colonic adenocarcinomas (AC). To date there are no data regarding E2F 1 expression and its relationship with tumour kinetics (proliferation, apoptosis) in adenocarcinomas that develop on Barrett oesophagus. AIM: As oesophageal adenocarcinomas occur almost exclusively in the metaplastic Barrett epithelium and the opposing E2F-1 behaviour seems to be cell and tissue-type dependent, we examined the manner in which E2F-1 acts in ACs of Barrett oesophagus. METHODS: We estimated the immunohistochemical expression of E2F-1, Ki-67, caspase-3 and p53 immunohistochemical status in 35 Barrett oesophagus ACs. RESULTS: E2F-1 immunopositivity correlated inversely with Ki-67, by semi-serial section and statistical analysis (p = 0.023, Spearman correlation). Semi-serial section analysis revealed a direct association between E2F-1 and caspase-3 staining. No correlation was found with p53 status. Cases with higher E2F-1 immunoexpression exhibited longer survival (p = 0.047, Cox-regression). CONCLUSIONS: E2F-1 expression was negatively related to tumour proliferation in ACs of Barrett oesophagus. Additionally, E2F-1 immunohistochemical status correlated positively with patient survival. These findings are opposite from those seen in OSCCs, suggesting that the tumour-suppressing E2F-1 behaviour in oesophageal adenocarcinomas is possibly due to the intestinal-type nature of the metaplastic Barrett mucosa. PMID- 17908805 TI - Hypoxia-inducible factor-1alpha expression correlates with focal macrophage infiltration, angiogenesis and unfavourable prognosis in urothelial carcinoma. AB - BACKGROUND: Hypoxia inducible factor (HIF)-1alpha is a critical regulatory protein of cellular response to hypoxia and is closely related to angiogenic process. AIMS: To explore the potential role and the prognostic value of HIF 1alpha in urothelial carcinoma (UC). METHODS: Clinicopathological and follow-up data on 99 UC cases were reviewed and immunostained for HIF-1alpha, CD68, vascular endothelial growth factor (VEGF) and CD34 antigen. Tumour-associated macrophage (TAM) counts and HIF-1alpha expression were compared with clinicopathologic characteristics, overall survival (OS) and disease-free survival rates (DFS). RESULTS: High expression of HIF-1alpha was detected in 55 of 99 (55.6%) tumours. HIF-1alpha expression was correlated with tumour size, histological grade, tumour invasiveness and recurrence. VEGF and microvessel density (MVD) demonstrated their positive correlation with HIF-1alpha overexpression, supporting the correlation of HIF-1alpha up-regulation with tumour angiogenesis. Higher TAM infiltration was identified in high expression of HIF-1alpha cases rather than HIF-1alpha low expression cases (p = 0.002). Kaplan Meier analysis found that HIF-1alpha overexpression and high TAM count was only associated with worse DFS (p = 0.009, p = 0.023) but was not associated with OS (p = 0.696, p = 0.141). Multivariate analyses indicated only tumour size (p = 0.038) to be an independently significant prognostic factor for OS, in addition, HIF-1alpha expression (p = 0.011), as well as histological grade (p = 0.038), and MVD (p = 0.004), to be independently significant prognostic factors for DFS. CONCLUSIONS: Our results indicate that HIF-1alpha is a key regulator of the angiogenic cascade. We show that HIF-1alpha is an independent prognostic factor for disease-free survival. PMID- 17908806 TI - Hybrid cell vaccination resolves Leishmania donovani infection by eliciting a strong CD8+ cytotoxic T-lymphocyte response with concomitant suppression of interleukin-10 (IL-10) but not IL-4 or IL-13. AB - There is an acute dearth of therapeutic interventions against visceral leishmaniasis that is required to restore an established defective cell-mediated immune response. Hence, formulation of effective immunotherapy requires the use of dominant antigen(s) targeted to elicit a specific antiparasitic cellular immune response. We implemented hybrid cell vaccination therapy in Leishmania donovani-infected BALB/c mice by electrofusing dominant Leishmania antigen kinetoplastid membrane protein 11 (KMP-11)-transfected bone marrow-derived macrophages from BALB/c mice with allogeneic bone marrow-derived dendritic cells from C57BL/6 mice. Hybrid cell vaccine (HCV) cleared the splenic and hepatic parasite burden, eliciting KMP-11-specific major histocompatibility complex class I-restricted CD8+ cytotoxic T-lymphocyte (CTL) responses. Moreover, splenic lymphocytes of HCV-treated mice not only showed the enhancement of gamma interferon but also marked an elevated expression of the Th2 cytokines interleukin-4 (IL-4) and IL-13 at both transcriptional and translational levels. On the other hand, IL-10 production from splenic T cells was markedly suppressed as a result of HCV therapy. CD8+ T-cell depletion completely abrogated HCV mediated immunity and the anti-KMP-11 CTL response. Interestingly, CD8+ T-cell depletion completely abrogated HCV-induced immunity, resulting in a marked increase of IL-10 but not of IL-4 and IL-13. The present study reports the first implementation of HCV immunotherapy in an infectious disease model, establishing strong antigen-specific CTL generation as a correlate of HCV-mediated antileishmanial immunity that is reversed by in vivo CD8+ T-cell depletion of HCV treated mice. Our findings might be extended to drug-nonresponsive visceral leishmaniasis patients, as well as against multiple infectious diseases with pathogen-specific immunodominant antigens. PMID- 17908808 TI - Contribution of Moraxella catarrhalis type IV pili to nasopharyngeal colonization and biofilm formation. AB - Moraxella catarrhalis is a gram-negative mucosal pathogen of the human respiratory tract. Although little information is available regarding the initial steps of M. catarrhalis pathogenesis, this organism must be able to colonize the human mucosal surface in order to initiate an infection. Type IV pili (TFP), filamentous surface appendages primarily comprised of a single protein subunit termed pilin, play a crucial role in the initiation of disease by a wide range of bacteria. We previously identified the genes that encode the major proteins involved in the biosynthesis of M. catarrhalis TFP and determined that the TFP expressed by this organism are highly conserved and essential for natural transformation. We extended this initial study by investigating the contribution of TFP to the early stages of M. catarrhalis colonization. TFP-deficient M. catarrhalis bacteria exhibit diminished adherence to eukaryotic cells in vitro. Additionally, our studies demonstrate that M. catarrhalis cells form a mature biofilm in continuous-flow chambers and that biofilm formation is enhanced by TFP expression. The potential role of TFP in colonization by M. catarrhalis was further investigated using in vivo studies comparing the abilities of wild-type M. catarrhalis and an isogenic TFP mutant to colonize the nasopharynx of the chinchilla. These results suggest that the expression of TFP contributes to mucosal airway colonization. Furthermore, these data indicate that the chinchilla model of nasopharyngeal colonization provides an effective animal system for studying the early steps of M. catarrhalis pathogenesis. PMID- 17908807 TI - Neutrophils play an important role in host resistance to respiratory infection with Acinetobacter baumannii in mice. AB - Acinetobacter baumannii has emerged as a major cause of both community-associated and nosocomial pneumonia, but little is known about the cellular and molecular mechanisms of host defense against respiratory infection with this bacterial pathogen. In this study, we examined the role of neutrophils in host resistance to pulmonary A. baumannii infection in a mouse model of intranasal (i.n.) infection. We found that neutrophils were rapidly recruited to the lungs following i.n. inoculation of the pathogen and declined to baseline level upon clearance of the infection. Depletion of neutrophils using monoclonal antibody RB6-8C5 prior to infection resulted in an acute lethal infection that was associated with enhanced bacterial burdens in the lung (P < 0.05) and extrapulmonary dissemination to the spleen. The increased susceptibility to A. baumannii in neutropenic mice was associated with a delay in the mRNA expression and production of early proinflammatory cytokines such as tumor necrosis factor alpha, interleukin-6, keratinocyte chemoattractant protein, monocyte chemoattractant protein 1, and macrophage inflammatory protein 2 (MIP-2) in the lungs and development of severe bronchopneumonia and lymphoid tissue destruction in the spleen. Moreover, i.n. administration of the neutrophil-inducing chemokine MIP-2 to normal mice induced a pulmonary influx of neutrophils and significantly enhanced the clearance of A. baumannii from the lungs (P < 0.01). These results imply that neutrophils play a critical role in host resistance to respiratory A. baumannii infection. PMID- 17908809 TI - Combination of protein and viral vaccines induces potent cellular and humoral immune responses and enhanced protection from murine malaria challenge. AB - The search for an efficacious vaccine against malaria is ongoing, and it is now widely believed that to confer protection a vaccine must induce very strong cellular and humoral immunity concurrently. We studied the immune response in mice immunized with the recombinant viral vaccines fowlpox strain FP9 and modified virus Ankara (MVA), a protein vaccine (CV-1866), or a combination of the two; all vaccines express parts of the same preerythrocytic malaria antigen, the Plasmodium berghei circumsporozoite protein (CSP). Mice were then challenged with P. berghei sporozoites to determine the protective efficacies of different vaccine regimens. Two immunizations with the protein vaccine CV-1866, based on the hepatitis B core antigen particle, induced strong humoral immunity to the repeat region of CSP that was weakly protective against sporozoite challenge. Prime-boost with the viral vector vaccines, FP9 followed by MVA, induced strong T cell immunity to the CD8+ epitope Pb9 and partially protected animals from challenge. Physically mixing CV-1866 with FP9 or MVA and then immunizing with the resultant combinations in a prime-boost regimen induced both cellular and humoral immunity and afforded substantially higher levels of protection (combination, 90%) than either vaccine alone (CV-1866, 12%; FP9/MVA, 37%). For diseases such as malaria in which different potent immune responses are required to protect against different stages, using combinations of partially effective vaccines may offer a more rapid route to achieving deployable levels of efficacy than individual vaccine strategies. PMID- 17908810 TI - Agonists of Toll-like receptors 3, 4, 7, and 9 are candidates for use as adjuvants in an outer membrane vaccine against Neisseria meningitidis serogroup B. AB - The bacterium Neisseria meningitidis is the causative agent of meningitis and sepsis. A generally effective vaccine against N. meningitidis serogroup B is not yet available, but outer membrane vesicle vaccines are in development. These vaccines contain lipopolysaccharide (LPS). The inclusion of N. meningitidis wild type LPS in a vaccine is controversial because of its high toxicity. Therefore, the adjuvant activity of a panel of different Toll-like receptor (TLR) agonists in combination with LPS-deficient meningococcal outer membrane complexes was compared after immunization of mice. The results demonstrate that TLR3, TLR4, TLR7, and TLR9 agonists enhance immune responses against LPS-deficient outer membrane complexes. Their adjuvant activity was characterized by higher levels of antigen-specific immunoglobulin G (IgG), IgG2a, and IgG2b; a higher IgG2a/IgG1 ratio; lower total IgE levels; and most importantly, higher serum bactericidal antibody titers compared to LPS-deficient outer membrane complexes alone. PMID- 17908811 TI - Immunoepidemiology of Wuchereria bancrofti infection: parasite transmission intensity, filaria-specific antibodies, and host immunity in two East African communities. AB - We compared the age profiles of infection and specific antibody intensities in two communities with different transmission levels in East Africa to examine the contribution of humoral responses to human immunity to the vector-borne helminth Wuchereria bancrofti. The worm intensities were higher and exhibited a nonlinear age pattern in a high-transmission community, Masaika, in contrast to the low but linearly increasing age infection profile observed for a low-transmission community, Kingwede. The mean levels of specific immunoglobulin G1 (IgG1), IgG2, IgG4, and IgE were also higher in Masaika, but intriguingly, the IgG3 response was higher in Kingwede. The age-antibody patterns differed in the two communities but in a manner apparently contrary to a role in acquired immunity when the data were assessed using simple correlation methods. By contrast, multivariate analyses showed that the antibody response to infection may be classified into three types and that two of these types, a IgG3-type response and a response measuring a trade-off in host production of IgG4 and IgG3 versus production of IgG1, IgG2, and IgE, had a negative effect on Wuchereria circulating antigen levels in a manner that supported a role for these responses in the generation of acquired immunity to infection. Mathematical modeling supported the conclusions drawn from empirical data analyses that variations in both transmission and worm intensity can explain community differences in the age profiles and impacts of these antibody response types. This study showed that parasite-specific antibody responses may be associated with the generation of acquired immunity to human filarial infection but in a form which is dependent on worm transmission intensity and interactions between immune components. PMID- 17908813 TI - Chlamydia trachomatis species-specific induction of ezrin tyrosine phosphorylation functions in pathogen entry. AB - Chlamydia trachomatis is an obligate intracellular pathogen of humans that exhibits species-specific biological characteristics in its early interactions with host cells that are likely important to pathogenesis. One such characteristic is the tyrosine phosphorylation (Tyr-P) of an approximately 70-kDa polypeptide that occurs only after infection of mammalian cells by human strains. We sought to identify this protein because of its potential significance to the pathogenesis of human chlamydial infections. Using an immunoproteomic approach we identified the host protein ezrin, a member of the ezrin-radixin-moesin (ERM) protein family that serves as a physical link between host cell receptors and the actin cytoskeleton. Confocal microscopy studies showed colocalization of ezrin and actin at the tips and crypts of microvilli, the site of chlamydial attachment and entry, respectively. To demonstrate a functional role for ezrin we infected cells with a dominant-negative (DN) ezrin phenotype or treated cells with ezrin specific small interfering RNA (siRNA). We found that both DN and siRNA-treated cells were significantly less susceptible to infection by human chlamydial strains. Moreover, we demonstrated that inhibition of infection in ezrin DN cells occurred at the stage of chlamydial entry. We hypothesize that the C. trachomatis specific Tyr-P of ezrin might relate to an undefined species-specific mechanism of pathogen entry that involves chlamydial specific ligand(s) and host cell coreceptor usage. PMID- 17908812 TI - The p38 mitogen-activated protein kinase signaling pathway is coupled to Toll like receptor 5 to mediate gene regulation in response to Pseudomonas aeruginosa infection in human airway epithelial cells. AB - In this study, we show that stimulation of human airway epithelial cells (HAECs) by Pseudomonas aeruginosa strain PAO1 induces time- and dose-dependent activation of p38 mitogen-activated protein kinase (MAPK). Activated p38 MAPK stayed in the cytoplasm instead of translocating to the nucleus, as shown by cellular fractionation. p38 MAPK was activated when HAECs were incubated with P. aeruginosa strain PAK and Burkholderia cepacia, while little activation was observed with the isogenic flagellin-free strains PAK/fliC and B. cepacia BC/fliC. The presence of Toll-like receptor 5 (TLR5) in 293 cells mediated PAO1 dependent activation of p38 MAPK, and in HAECs p38 MAPK activation was blocked by the overexpression of a dominant negative TLR5. Two inhibitors of p38 MAPK, SB202190 and SB203580, significantly attenuated PAO1-dependent expression of an NF-kappaB-dependent luciferase reporter gene, suggesting that p38 MAPK activation is required for full activation of NF-kappaB-dependent signaling. Microarray analysis of NF-kappaB target genes revealed up-regulation of multiple genes by PAO1 in HAECs. Reverse transcription-PCR and protein expression analysis were used to show that up-regulation of NF-kappaB-dependent genes induced by PAO1, such as the genes encoding Cox-2 and interleukin-8, was attenuated by SB203580. These results demonstrate a role for p38 MAPK signaling in gene regulation in response to P. aeruginosa via TLR5. PMID- 17908814 TI - Phosphoinositide-3-kinase-dependent, MyD88-independent induction of CC-type chemokines characterizes the macrophage response to Toxoplasma gondii strains with high virulence. AB - Chemokines play an important role in inflammation and infection due to their ability to recruit cells of innate and adaptive immunity. Here we examined mouse macrophage chemokine responses during intracellular infections with high- and low virulence Toxoplasma gondii strains. The high-virulence type I strain RH induced a large panel of CC-type chemokines, whereas responses elicited by strains PTG (type II) and M7741 (type III) were much weaker. Strikingly, the T. gondii induced chemokine response occurred independently of signaling through the Toll like receptor adaptor MyD88. Instead, production of chemokines during infection was heavily dependent upon phosphoinositide-3-kinase signaling pathways. Because infection with type I strains such as RH results in an uncontrolled proinflammatory cytokine response, we hypothesize that this virulence phenotype is a consequence of early strong induction of chemokines by type I, but not type II or III, Toxoplasma strains. PMID- 17908816 TI - Distortion-corrected T2 weighted MRI: a novel approach to prostate radiotherapy planning. AB - The purpose of this study was to evaluate distortion-corrected MRI as a radiotherapy planning tool for prostate cancer and the resultant implications for dose sparing of organs at risk. 11 men who were to be treated with radical conformal radiotherapy for localized prostate cancer had an MRI scan under radiotherapy planning conditions, which was corrected for geometric distortion. Radiotherapy plans were created for planning target volumes derived from the MRI- and CT-defined prostate. Dose volume histograms were produced for the rectum, bladder and penile bulb. The mean volume of the prostate as defined on CT and MRI was 41 cm3 and 36 cm3, respectively (p = 0.009). The predicted percentage of the rectum treated to dose levels of 45-65 Gy was significantly lower for plans delineating the prostate with MRI than for those with CT. The rectal-sparing effect was confined to the lowermost 4 cm of the rectum (anal canal). There were no differences between the predicted doses to bladder or penile bulb (as defined using MRI) between plans. In conclusion, prostate radiotherapy planning based on distortion-corrected MRI is feasible and results in a smaller target volume than does CT. This leads to a lower predicted proportion of the rectum, in particular the lower rectum (anal canal), treated to a given dose than with CT. PMID- 17908815 TI - Chlamydia pneumoniae inclusion membrane protein Cpn0585 interacts with multiple Rab GTPases. AB - Chlamydiae are intracellular bacteria that develop within a membrane-bound vacuole called an inclusion. To ensure that the inclusion is a safe niche for chlamydial replication, chlamydiae exploit a number of host cell processes, including membrane-trafficking pathways. Recently, several Rab GTPases were found to associate with the inclusions of various chlamydial species. Here we report that Cpn0585, a Chlamydia pneumoniae inclusion membrane protein (Inc), interacts with multiple Rab GTPases. The results from yeast two-hybrid experiments revealed that an amino-terminally truncated form of Cpn0585 (Cpn0585(102-651)) interacts with Rab1, Rab10, and Rab11 but not with Rab4 or Rab6. Cpn0585-Rab GTPase interactions are direct and GTP dependent as shown in glutathione S-transferase pull-down assays using native and recombinant Cpn0585. In C. pneumoniae-infected HEp-2 cells transfected with enhanced green fluorescent protein (EGFP)-tagged Rab GTPases, the colocalization with Cpn0585 at the inclusion membrane was partial for EGFP-Rab1 and EGFP-Rab10, but extensive for wild-type EGFP-Rab11A and the constitutively active GTPase-deficient EGFP-Rab11AQ70L. Moreover, Cpn0585 colocalized with EGFP-Rab11AQ70L as early as 2 h postinfection. Upon delivery into live C. pneumoniae-infected cells, Cpn0585(628-651)-specific antibodies bound to the inclusion membrane, demonstrating that the Rab GTPase-interacting domain of Cpn0585 faces the host cell cytosol. Finally, ectopic expression of Cpn0585(102-651) partially inhibited the development of C. pneumoniae inclusions in EGFP. but not in EGFP-Rab11AQ70L-expressing HEp-2 cells. Collectively, these data suggest that Cpn0585 is involved in the recruitment of Rab GTPases to the inclusion membrane and that interfering with this function may adversely impact the fitness of the C. pneumoniae inclusion for chlamydial replication. PMID- 17908817 TI - The CT appearances of gallbladder perforation. AB - Perforation of the gallbladder is an uncommon complication of acute cholecystitis that is associated with relatively high mortality. Symptoms and clinical signs can be indistinguishable from those of uncomplicated acute cholecystitis, leading to delayed diagnosis. We reviewed the clinical and imaging findings in 17 patients with gallbladder perforation confirmed at surgery. PMID- 17908818 TI - Hypoxia can be detected in irradiated normal human tissue: a study using the hypoxic marker pimonidazole hydrochloride. AB - Chronic tissue hypoxia may play a role in the pathogenesis of late radiation fibrosis. In order to investigate this hypothesis, the immunohistochemical distribution of pimonidazole hydrochloride (n = 14 patients) and carbonic anhydrase IX (CAIX) (n = 38 patients) was studied in samples of previously irradiated normal human tissue. One sample of irradiated breast tissue, which also showed marked histological features of radiation injury, stained positive for pimonidazole hydrochloride. No CAIX staining was seen in irradiated tissue other than some evidence of physiological hypoxia in the epidermis of two samples of irradiated skin; both were positive for pimonidazole and one was focally positive for CAIX. Pimonidazole hydrochloride staining of tissue with morphological changes of radiation injury could support a role for hypoxia in the pathogenesis of late normal tissue fibrosis in humans. PMID- 17908819 TI - Imaging findings of radiation-induced sarcoma of the head and neck. AB - We set out to retrospectively review the clinical and imaging features of patients with post-radiation sarcoma, especially in the head and neck region. We reviewed the records of 4194 patients with carcinoma of the head and neck region who had a history of radiation. They had undergone CT and/or MRI. Medical records were reviewed for the primary diagnosis, radiation history and latency period to the development of sarcoma. The patients included four men and two women with a mean age of 64.5 years. The mean latency period for the development of sarcoma was 11.5 years. Primary diagnoses were maxillary carcinoma, nasopharyngeal carcinoma, adenoid cystic carcinoma of the oral floor, tonsilar carcinoma, soft palate carcinoma and tongue carcinoma. Histopathological examinations revealed osteosarcoma, spindle cell sarcoma, chondrosarcoma, malignant peripheral nerve sheath tumour, spindle cell carcinoma and malignant fibrous histiocytoma, respectively. Common findings were a heterogeneous and well-enhanced soft tissue mass and bone destruction. There is at present little or no prospect for the effective prevention of radiation-induced sarcoma of the head and neck. This emphasizes the importance of the earliest possible diagnosis for such patients. The imaging findings are not diagnosis specific, but strict follow-up within the radiation field by CT and MRI and an appreciation of the expected latency period may help to provide the diagnosis. When radiotherapy is performed for head and neck neoplasms, periodic follow-up observations may be necessary for many years. PMID- 17908820 TI - Lower limb contrast venography: a modified technique for use in thromboprophylaxis clinical trials for the accurate evaluation of deep vein thrombosis. AB - Lower limb venography remains the imaging modality of choice for detection of asymptomatic deep vein thrombosis (DVT) in clinical trials of anticoagulant agents. A variety of techniques of venography have been described. Here, we describe a modified technique (the "King's" technique) developed to increase the overall adequacy of identification of lower limb veins and detection of small asymptomatic DVT. Essential elements include proper preparation of patients prior to their arrival in the radiology department, intermittent use of tourniquets to ensure complete and adequate deep vein filling, use of a consistent image acquisition sequence and visualization of all veins in at least two different planes. Use of this technique minimizes technical difficulties, provides improved patient through-put in "busy" fluoroscopy units and, ultimately, improves "off site" levels of adjudication. PMID- 17908821 TI - A comprehensive ChIP-chip analysis of E2F1, E2F4, and E2F6 in normal and tumor cells reveals interchangeable roles of E2F family members. AB - Using ChIP-chip assays (employing ENCODE arrays and core promoter arrays), we examined the binding patterns of three members of the E2F family in five cell types. We determined that most E2F1, E2F4, and E2F6 binding sites are located within 2 kb of a transcription start site, in both normal and tumor cells. In fact, the majority of promoters that are active (as defined by TAF1 or POLR2A binding) in GM06990 B lymphocytes and Ntera2 carcinoma cells were also bound by an E2F. This very close relationship between E2F binding sites and binding sites for general transcription factors in both normal and tumor cells suggests that a chromatin-bound E2F may be a signpost for active transcription initiation complexes. In general, we found that several E2Fs bind to a given promoter and that there is only modest cell type specificity of the E2F family. Thus, it is difficult to assess the role of any particular E2F in transcriptional regulation, due to extreme redundancy of target promoters. However, Ntera2 carcinoma cells were exceptional in that a large set of promoters were bound by E2F6, but not by E2F1 or E2F4. It has been proposed that E2F6 contributes to gene silencing by recruiting enzymes involved in methylating histone H3. To test this hypothesis, we created Ntera2 cell lines harboring shRNAs to E2F6. We found that reduction of E2F6 only induced minimal alteration of the transcriptome of Ntera2 transcriptome. Our results support the concept of functional redundancy in the E2F family and suggest that E2F6 is not critical for histone methylation. PMID- 17908823 TI - SHARCGS, a fast and highly accurate short-read assembly algorithm for de novo genomic sequencing. AB - The latest revolution in the DNA sequencing field has been brought about by the development of automated sequencers that are capable of generating giga base pair data sets quickly and at low cost. Applications of such technologies seem to be limited to resequencing and transcript discovery, due to the shortness of the generated reads. In order to extend the fields of application to de novo sequencing, we developed the SHARCGS algorithm to assemble short-read (25-40-mer) data with high accuracy and speed. The efficiency of SHARCGS was tested on BAC inserts from three eukaryotic species, on two yeast chromosomes, and on two bacterial genomes (Haemophilus influenzae, Escherichia coli). We show that 30-mer based BAC assemblies have N50 sizes >20 kbp for Drosophila and Arabidopsis and >4 kbp for human in simulations taking missing reads and wrong base calls into account. We assembled 949,974 contigs with length >50 bp, and only one single contig could not be aligned error-free against the reference sequences. We generated 36-mer reads for the genome of Helicobacter acinonychis on the Illumina 1G sequencing instrument and assembled 937 contigs covering 98% of the genome with an N50 size of 3.7 kbp. With the exception of five contigs that differ in 1 4 positions relative to the reference sequence, all contigs matched the genome error-free. Thus, SHARCGS is a suitable tool for fully exploiting novel sequencing technologies by assembling sequence contigs de novo with high confidence and by outperforming existing assembly algorithms in terms of speed and accuracy. PMID- 17908822 TI - Altered adenosine-to-inosine RNA editing in human cancer. AB - Adenosine-to-inosine (A-to-I) RNA editing was recently shown to be abundant in the human transcriptome, affecting thousands of genes. Employing a bioinformatic approach, we identified significant global hypoediting of Alu repetitive elements in brain, prostate, lung, kidney, and testis tumors. Experimental validation confirmed this finding, showing significantly reduced editing in Alu sequences within MED13 transcripts in brain tissues. Looking at editing of specific recoding and noncoding sites, including in cancer-related genes, a more complex picture emerged, with a gene-specific editing pattern in tumors vs. normal tissues. Additionally, we found reduced RNA levels of all three editing mediating enzymes, ADAR, ADARB1, and ADARB2, in brain tumors. The reduction of ADARB2 correlated with the grade of malignancy of glioblastoma multiforme, the most aggressive of brain tumors, displaying a 99% decrease in ADARB2 RNA levels. Consistently, overexpression of ADAR and ADARB1 in the U87 glioblastoma multiforme cell line resulted in decreased proliferation rate, suggesting that reduced A-to-I editing in brain tumors is involved in the pathogenesis of cancer. Altered epigenetic control was recently shown to play a central role in oncogenesis. We suggest that A-to-I RNA editing may serve as an additional epigenetic mechanism relevant to cancer development and progression. PMID- 17908825 TI - Using buprenorphine for outpatient opioid detoxification. AB - The Drug Addiction Treatment Act of 2000 (DATA 2000) was established to create a new paradigm for medication-assisted treatment of persons with opiate addiction in the United States. Before enactment of DATA 2000, the use of opioid medications to treat patients with opioid addiction was permissible only in federally approved treatment programs, ie, "methadone clinics." The only medications permitted were Schedule II drugs (eg, methadone hydrochloride and l alpha-acetylmethadol [LAAM]), which could only be dispensed, not prescribed. Under provisions of DATA 2000, qualified physicians in a medical office and other appropriate settings outside the opioid treatment program system may prescribe and/or dispense Schedule III, IV, and V opioid medications for treating persons with opioid addiction if such medications have been specifically approved by the US Food and Drug Administration for that indication. Opioid addiction treatment programs were commonly known as methadone clinics. Such programs now may also dispense buprenorphine hydrochloride and the buprenorphine hydrochloride-naloxone combination. PMID- 17908824 TI - Reductive evolution of architectural repertoires in proteomes and the birth of the tripartite world. AB - The repertoire of protein architectures in proteomes is evolutionarily conserved and capable of preserving an accurate record of genomic history. Here we use a census of protein architecture in 185 genomes that have been fully sequenced to generate genome-based phylogenies that describe the evolution of the protein world at fold (F) and fold superfamily (FSF) levels. The patterns of representation of F and FSF architectures over evolutionary history suggest three epochs in the evolution of the protein world: (1) architectural diversification, where members of an architecturally rich ancestral community diversified their protein repertoire; (2) superkingdom specification, where superkingdoms Archaea, Bacteria, and Eukarya were specified; and (3) organismal diversification, where F and FSF specific to relatively small sets of organisms appeared as the result of diversification of organismal lineages. Functional annotation of FSF along these architectural chronologies revealed patterns of discovery of biological function. Most importantly, the analysis identified an early and extensive differential loss of architectures occurring primarily in Archaea that segregates the archaeal lineage from the ancient community of organisms and establishes the first organismal divide. Reconstruction of phylogenomic trees of proteomes reflects the timeline of architectural diversification in the emerging lineages. Thus, Archaea undertook a minimalist strategy using only a small subset of the full architectural repertoire and then crystallized into a diversified superkingdom late in evolution. Our analysis also suggests a communal ancestor to all life that was molecularly complex and adopted genomic strategies currently present in Eukarya. PMID- 17908826 TI - Eliminating disparities in pain management. AB - Not all patients are treated equally for their pain with some therefore being undertreated. Discrepancies still exist in the way physicians treat special populations of patients such as racial minorities, women, and substance abusers. All healthcare providers need to be aware of the not so readily apparent disparities resulting from stereotyping, bias, ageism, and socioeconomic considerations. Physicians can best provide appropriate and equal care by following pain management guidelines; however, they may receive contradictory information and be apprehensive about prescribing opioids, especially to substance abusers. In this "refreshed" article, the authors describe patient encounters with patients of color and offer some goals for removing inequality and inequity from clinical settings. PMID- 17908827 TI - What is being done to address the new drug epidemic? AB - As osteopathic physicians care for patients with complaints of pain, they commonly prescribe controlled substances. The use of these agents presents special challenges for providers, patients, and communities. The US Drug Enforcement Administration (DEA) has provided testimony to the US Congress in regard to the growing problem of diversion and misuse of such medications. Joseph T. Rannazzisi, the deputy assistant administrator in the Office of Diversion Control, appeared before the House Government Reform Committee's Subcommittee on Criminal Justice, Drug Policy, and Human Resources on July 26, 2006. PMID- 17908829 TI - Using opioids for patients with moderate to severe pain. AB - In the United States, many visits to physician offices are for complaints of pain. Patients who have moderate to severe pain can be effectively treated with different modalities, including opioids. Proper management requires that physicians be open-minded and thorough. Physicians should take a complete history and perform a complete physical evaluation including an osteopathic structural examination to develop a comprehensive treatment plan. This strategy should include follow-up visits for continued assessment of therapy. Continued reassessment of treatment and patient responsiveness have been shown to be most beneficial to both physician and patient. Using the osteopathic medical model of treatment, physicians should identify psychosocial as well as somatic dysfunctions and appropriately treat patients for them. They should not avoid prescribing opioids because of fear of a patient's becoming addicted, but instead, integrate the use of such important analgesics in a multidisciplinary treatment plan. However, it must be recognized that opioids are powerful medications that require monitoring and dosing according to patient response. PMID- 17908828 TI - Improving physician and medical student education in substance use disorders. AB - Medical and psychosocial problems related to substance use disorders (SUDs) remain a major source of national morbidity and mortality. This situation exists despite greater understanding of genetic, neurobiologic, and social underpinnings of the development of these illnesses that has resulted in many advances in addiction medicine. The value of assessment and brief intervention of this disease is well documented. Patients need to be identified and engaged in order for them to be treated. A variety of evidence-based pharmacologic and psychotherapeutic treatments are now available. Strong evidence exists that treatment of patients for SUDs produces results similar to or better than those obtained from treatment for other chronic illnesses. It is also clear that physicians can play a pivotal role in helping to reduce the burden of disease related to SUDs However, to do this, physicians need to be better educated. Through such education comes greater confidence in identification and providing treatment. Also, the discomfort and stigma often associated with this disease are reduced. The federal government-through the Office of National Drug Control Policy, the Surgeon General, the Center for Substance Abuse Treatment, the National Institute on Drug Abuse, the National Institute on Alcohol Abuse and Alcoholism, and the National Highway Traffic Safety Administration of the Department of Transportation (DOT)-is expending concerted efforts to improve physician education in addiction medicine. These efforts culminated in the Second Leadership Conference on Medical Education in Substance Abuse in December 2006. The osteopathic medical profession was represented at this conference. This article reviews not only the recommendations from this meeting, but also the nature of the problem, how members of the osteopathic medical profession are currently addressing it, and a strategy for improvement endorsed by the American Osteopathic Academy of Addiction Medicine. PMID- 17908830 TI - Overactive bladder: treatment options in primary care medicine. AB - Overactive bladder is a highly prevalent condition, affecting approximately 33 million adults in the United States. Despite the considerable impact this condition has on patients' quality of life, overactive bladder remains underrecognized and undertreated as a result of patient embarrassment and reluctance to seek medical help, as well as a lack of proactive questioning by physicians. The present article encourages physicians to initiate a dialogue with patients regarding urinary control and, specifically, overactive bladder. Treatment options for overactive bladder recommended in the current article include both nonpharmacologic and pharmacologic therapies. Properties of antimuscarinic agents, including three new drug therapies, are reviewed and provided for physicians to optimize therapy options, particularly among elderly patients. PMID- 17908831 TI - Role of osteopathic manipulative treatment in altering pain biomarkers: a pilot study. AB - CONTEXT: Underlying mechanisms explaining the effects of osteopathic manipulative treatment (OMT) are poorly defined. The authors evaluate various nociceptive (pain) biomarkers that have been suggested as important mediators in this process. OBJECTIVE: To determine if OMT influences levels of circulatory pain biomarkers. METHODS: In a prospective, blinded assessment, blood was collected from 20 subjects (10 with chronic low back pain [LBP], 10 controls without chronic LBP) for 5 consecutive days. On day 4, OMT was administered to subjects 1 hour before blood collection. Blood was analyzed for levels of beta-endorphin (betaE), serotonin (5-hydroxytryptamine [5-HT]), 5-hydroxyindoleacetic acid (5 HIAA), anandamide (arachidonoylethanolamide [AEA]), and N-palmitoylethanolamide (PEA). A daily questionnaire was used to monitor confounding factors, including pain and stress levels, sleep patterns, and substance use. RESULTS: Increases from baseline in betaE and PEA levels and a decrease in AEA levels occurred immediately posttreatment. At 24 hours posttreatment, similar biomarker changes from baseline were observed. A decrease in stress occurred from baseline to day 5. The change in PEA from baseline to 24 hours posttreatment correlated with the corresponding changes in stress. Subgroup analysis showed that subjects with chronic LBP had significantly reduced 5-HIAA levels at 30 minutes posttreatment (P=.05) and 5-HT levels at 24 hours posttreatment (P=.02) when compared with baseline concentrations. The increase in PEA in subjects with chronic LBP at 30 minutes posttreatment was two times greater than the increase in control subjects. CONCLUSION: Concentrations of several circulatory pain biomarkers were altered after OMT. The degree and duration of these changes were greater in subjects with chronic LBP than in control subjects without the disorder. PMID- 17908832 TI - Gender differences and alcohol use in the US Army. AB - CONTEXT: Operation Iraqi Freedom offered an opportunity to study the role of alcohol use among men and women serving in the US Army. The goal of this study was to determine whether there are gender-based differences in alcohol use among US Army soldiers, and if so, to evaluate the role of alcohol education efforts in the military. METHODS: In February 2005, 1200 individuals enlisted in the US Army were asked to complete a 29-item questionnaire regarding alcohol-use patterns. Survey topics included attitudes toward alcohol consumption and associated negative consequences. RESULTS: Six hundred eighty-five men and 325 women (N=1010) responded to the questionnaire for an overall response rate of 84%. Although men were more likely to engage in "bolus" drinking (ie, binge drinking), women exceeded established guidelines for safe alcohol consumption at a risk adjusted rate nearly twice that of men. In addition, for individuals whose behaviors were not in conformity with public health guidelines for safe alcohol consumption, the severity of reported negative consequences was influenced by gender. Women initially experience greater psychosocial impairment, and-should harmful drinking patterns progress to alcohol dependency-they are at greater risk of injury, morbidity, and mortality than men. CONCLUSION: Several gender-specific differences in alcohol-consumption patterns were found. Because the present study also found that women generally have more interest in educational interventions for alcohol abuse issues, however, researchers conclude that the efficacy of US Army risk-reduction programs would be improved by addressing gender-based differences. PMID- 17908833 TI - Trauma-induced pneumothorax after "bear-hug back crack" home remedy: attempted spinal manipulation by a layperson. AB - Spinal manipulation, as practiced by US-trained osteopathic physicians, is a safe and effective method of resolving patient pain and encouraging desirable physiologic improvement--often without pharmacologic intervention. Though novices, laypeople, and other clinicians also use manual techniques with similar goals in mind, their results are varied and sometimes dangerous to those they would help. The authors describe a case in which a layperson attempted spinal manipulation on a 20-year-old woman who later required a chest tube thoracostomy and hospitalization as a result of a pneumothorax. Osteopathic physicians are encouraged to consider patient risk factors for pneumothorax as a contraindication for the use of thoracic thrust techniques. PMID- 17908834 TI - Chronic psoas syndrome caused by the inappropriate use of a heel lift. AB - Heel lifts are commonly recommended for patients to manage the pain and discomfort of leg length discrepancies. However, used inappropriately, orthotics can create additional pain instead of alleviating it. In the case described, a 79 year-old male physician used a recommended heel lift for a perceived leg length discrepancy after right hip arthroplasty. Six months postsurgery, chronic, intractable pain developed in his hip and groin. He underwent a battery of tests to locate the pain, but its source remained elusive. Osteopathic evaluation and radiographic examination revealed an absence of leg length discrepancy and the presence of chronic psoas syndrome. Osteopathic manipulative treatment was prescribed and heel lift therapy discontinued, and the patient reported complete remission from pain. PMID- 17908835 TI - Ethical considerations in publishing research involving human subjects. PMID- 17908836 TI - Nationalized dentistry: a fork in the road. PMID- 17908837 TI - Discounting fees. PMID- 17908838 TI - Oral, nonoral disease. PMID- 17908839 TI - Alveolar ridge keratosis. PMID- 17908841 TI - 'Lab on a chip' probes cells for common sign of oral cancer. PMID- 17908845 TI - The dental care of U.S. children: access, use and referrals by nondentist providers, 2003. AB - BACKGROUND: Improvements in oral health care services have not reached evenly across every segment of American society. The authors examine the role of nondentist practitioners in referring child patients for dental care by analyzing data from the 2003 Medical Expenditure Panel Survey conducted by the Agency for Healthcare Research and Quality and the National Center for Health Statistics. METHODS: The authors provide national estimates of the percentage of the civilian noninstitutionalized population of the United States aged 2 through 17 years who had a dental visit, who had a dental checkup and who received advice from a nondentist health care provider to have a dental checkup. RESULTS: Overall, 38 percent of all poor, near-poor or low-income children and 60 percent of all middle- or high-income children aged 2 through 17 years reported having had a dental checkup during 2003. The authors observed no significant differences between poor, near-poor and low-income children and higher-income children in terms of having been advised by a nondentist health care provider to have a dental checkup. CONCLUSION: Although income may not predict the likelihood of patients' receiving advice from a nondentist health care provider to have a dental checkup, children from families with higher levels of income were more likely to seek dental care than were children from families with lower levels of income. Practice Implications. Efforts to increase access to dental care should aim to maximize the benefit of advice provided by nondentist health care practitioners to receive a dental checkup, so that children from families with limited income are as likely to receive a dental checkup as are children from families with higher levels of income. PMID- 17908844 TI - Tooth loss, dementia and neuropathology in the Nun study. AB - BACKGROUND: Numerous studies have linked dementia to the subsequent deterioration of oral health. Few investigators, however, have examined oral disease as a potential risk factor in the development of dementia. The authors conducted a study to investigate a potential association between a history of oral disease and the development of dementia. METHODS: Longitudinal dental records supplemented data collected from 10 annual cognitive assessments of 144 Milwaukee participants in the Nun Study, a longitudinal study of aging and Alzheimer disease, who were 75 to 98 years old. Neuropathologic findings at autopsy were available for 118 participants who died. RESULTS: A low number of teeth increased the risk of higher prevalence and incidence of dementia. CONCLUSION: Participants with the fewest teeth had the highest risk of prevalence and incidence of dementia. CLINICAL IMPLICATIONS: Edentulism or very few (one to nine) teeth may be predictors of dementia late in life. PMID- 17908846 TI - The use of compound topical anesthetics: a review. AB - BACKGROUND: The author reviewed the history of, federal regulations regarding, risks of and adverse drug reactions of five compound topical anesthetics: tetracaine, adrenaline/epinephrine and cocaine (TAC); lidocaine, adrenaline/epinephrine and tetracaine (LET); lidocaine, tetracaine and phenylephrine (TAC 20 percent Alternate); lidocaine, prilocaine and tetracaine (Profound); and lidocaine, prilocaine, tetracaine and phenylephrine with thickeners (Profound PET). TYPES OF STUDIES REVIEWED: The author reviewed clinical trials, case reports, descriptive articles, and U.S. Food and Drug Administration (FDA) regulations and recent public advisory warnings regarding the federal approval of and risks associated with the use of compound topical anesthetics. RESULTS: Compound topical anesthetics are neither FDA-regulated nor unregulated. Some compounding pharmacies bypass the new FDA drug approval process, which is based on reliable scientific data and ensures that a marketed drug is safe, effective, properly manufactured and accurately labeled. Two deaths have been attributed to the lay use of compound topical anesthetics. In response, the FDA has announced the strengthening of its efforts against unapproved drug products. CLINICAL IMPLICATIONS: Compound topical anesthetics may be an effective alternative to local infiltration for some minimally invasive dental procedures; however, legitimate concerns exist in regard to their safety. Until they become federally regulated, compound topical anesthetics remain unapproved drug products whose benefits may not outweigh their risks for dental patients. PMID- 17908847 TI - Exposed bone in the palate. PMID- 17908848 TI - The incidence of four canals in maxillary first molars: a clinical determination. AB - BACKGROUND: The authors conducted an in vivo study to report the incidence of fourth root canals located and treated in maxillary first molars during a seven month period in a postgraduate endodontic program. METHODS: In this retrospective study, the authors determined the number of canals treated by postgraduate students in an endodontic program. The attending postgraduate endodontic faculty member supervising the case verified the number of canals in the teeth. The authors then collected the data from each resident and compiled them. RESULTS: The residents treated a total of 121 maxillary molars, 85 (70.2 percent) of which met the criterion of having four or more canals treated. Approximately 99 percent of the fourth canals were located in the mesiobuccal root. CONCLUSIONS: Overall, 70 percent of the maxillary first molars contained at least four canals that required instrumentation. CLINICAL IMPLICATIONS: The results of this study demonstrate that it is imperative for any dentist performing root canal therapy on maxillary first molars to examine carefully the pulpal floor to locate all canals, especially the second mesiobuccal canal. Performing thorough examinations may increase the chance of treatment success. PMID- 17908849 TI - Surface antibacterial properties of glass ionomer cements used in atraumatic restorative treatment. AB - BACKGROUND: Atraumatic restorative treatment (ART) is recommended for use worldwide, not only in developing countries where resources are not readily available, but also in more industrialized countries. The antibacterial properties of restorative dental materials may improve the restorative treatment outcome. Glass ionomer cement (GIC) has been advocated as the preferred restoration material for ART. The authors evaluated the antibacterial properties of restorative materials-three GICs and a zinc oxide eugenol (ZOE)-in vitro. METHODS: Streptococcus mutans, Actinomyces viscosus and Enterococcus faecalis were the test microorganisms. The authors used a quantitative microtiter spectrophotometric assay to evaluate the antibacterial effect of the restorative materials using the direct contact test (DCT) of freshly prepared and one-week aged materials. RESULTS: The freshly prepared GICs and ZOE showed no bacterial growth in all tested bacteria compared with a control. This effect lasted for at least one week for S. mutans and A. viscosus but not for E. faecalis. CONCLUSIONS: Conventional GICs used in ART showed antibacterial surface properties against cariogenic bacteria for at least one week. Further study on the long-term antimicrobial effects of GICs is needed. CLINICAL IMPLICATIONS: The antimicrobial properties of freshly prepared restorative materials and aged restorative materials used in ART have a potent effect against cariogenic bacteria. These properties have crucial importance in preventing secondary caries. PMID- 17908850 TI - A comparison of older and newer versions of intraoral digital radiography systems: diagnosing noncavitated proximal carious lesions. AB - BACKGROUND: The authors conducted a study to compare the accuracy of an older and newer version of two intraoral digital systems in terms of radiographic detection of proximal carious lesions. METHODS: Under in vitro and standardized conditions, the authors obtained radiographs of 160 noncavitated proximal surfaces using the Digora FMX (Soredex, Tuusula, Finland), the Digora Optime, the Schick CDR (Schick Technologies, Long Island City, N.Y.) and the Schick CDR Wireless (Schick Technologies) systems. Eight observers recorded proximal carious lesions on a five-point confidence scale. The presence of caries was validated histologically. RESULTS: The new digital systems (Digora Optime and Schick CDR Wireless) had significantly higher sensitivities than their predecessors. The authors found no significant differences in specificity among the Digora FMX, Schick CDR and Schick CDR Wireless systems, all of which had a significantly higher specificity than did the Digora Optime system (P < .02). The positive predictive value for the Digora Optime system was affected by its high sensitivity and low specificity, and it was lower than that for the two CDR systems (P < .02). CONCLUSIONS: Regarding overall accuracy, the difference between the older and newer versions of the photostimulable storage phosphor and complementary metal oxide semiconductor systems was not statistically significant. However, the authors found more false-positive diagnoses made with the Digora Optime system than with the Digora FMX system. CLINICAL IMPLICATIONS: Though the difference in specificities was statistically significant, the authors question whether the difference between the Digora Optime and the other systems is clinically relevant. Therefore, dentists can purchase any of these systems after considering factors other than those evaluated in this study. PMID- 17908852 TI - Strategic partnerships between academic dental institutions and communities: addressing disparities in oral health care. AB - BACKGROUND: A landmark report from the U.S. surgeon general identified disparities in oral health care as an urgent and high-priority problem. A parallel development in the dental education community is the growing consensus that significant curriculum reform is long overdue. METHODS: The authors performed a literature review and conducted a series of structured interviews with key institutional and community stakeholders from seven geographical regions of the United States. They investigated a wide range of partnerships between community-based dental clinics and academic dental institutions. RESULTS: On the basis of their interviews and literature review, the authors identified common themes and made recommendations to the dental community to improve access to care while enhancing the dental curriculum. CONCLUSIONS: Reducing disparities in access to oral health care and the need for reform of the dental curriculum may be addressed, in part, by a common solution: strategic partnerships between academic dental institutions and communities. Practice Implications. Organized dentistry and individual practitioners, along with other major stakeholders, can play a significant role in supporting reform of the dental curriculum and improving access to care. PMID- 17908851 TI - The in vivo contamination of air-driven low-speed handpieces with prophylaxis angles. AB - BACKGROUND: The authors conducted an in vivo study to determine if low-speed handpiece motors can become contaminated with oral flora when used with prophylaxis angles. METHODS: This crossover study involved 20 subjects, two types of handpieces and three prophylaxis angles. The authors used each handpiece/prophylaxis angle system to polish teeth. They then collected samples, spiral-plated the specimens and incubated them at 37 degrees C anaerobically and aerobically (with 5 percent carbon dioxide). After incubation, the authors examined the plates for the presence of bacterial colonies. RESULTS: At least 75 percent of the handpiece/prophylaxis angle systems used on the 20 subjects had bacterial contamination for at least one cultured area. Of the 420 specimens, 258 (61.4 percent) produced bacterial growth. Contamination varied from zero to 6,300 colony-forming units per milliliter. CONCLUSIONS: These data suggest that the internal surfaces of low-speed handpieces can become microbially contaminated during use with prophylaxis angles. CLINICAL IMPLICATIONS: Unless low-speed handpieces are sterilized properly after each use, they pose a risk for crossinfection. PMID- 17908853 TI - Frequently encountered errors in tooth preparations for crowns. PMID- 17908854 TI - Delegating tasks and growing the dental team. PMID- 17908855 TI - Is it ethical for dentists to treat their own children? PMID- 17908856 TI - For the dental patient. Gingival recession: causes and treatment. PMID- 17908857 TI - The electronic medical record. PMID- 17908858 TI - Seizures in children: determining the variation. PMID- 17908859 TI - Back to basics: congenital nephrogenic diabetes insipidus. PMID- 17908860 TI - Complementary, holistic, and integrative medicine: colic. PMID- 17908861 TI - Pediatrics in the community: Cap4Kids.org: connecting pediatricians to the community at the speed of light. PMID- 17908862 TI - Index of suspicion. PMID- 17908863 TI - Question from the clinician: alternating acetaminophen and ibuprofen in the treatment of fever. PMID- 17908864 TI - Firearms. PMID- 17908865 TI - Pharmacokinetics. PMID- 17908867 TI - Novel methods for diagnosis and treatment of posterolateral rotatory instability of the knee. PMID- 17908868 TI - The treatment of severe posttraumatic arthritis of the ankle joint. PMID- 17908869 TI - Hip arthroscopy in the athletic patient: current techniques and spectrum of disease. PMID- 17908870 TI - Management of proximal humeral fractures based on current literature. PMID- 17908871 TI - Functional problems and arthrofibrosis following total knee arthroplasty. PMID- 17908872 TI - The current state of cervical and lumbar spinal disc arthroplasty. PMID- 17908873 TI - Evaluation of complications associated with six hundred mini-subvastus total knee arthroplasties. PMID- 17908874 TI - Correction of hallux valgus using lateral soft-tissue release and proximal Chevron osteotomy through a medial incision. PMID- 17908875 TI - Posterior-stabilized constrained total knee arthroplasty for complex primary cases. PMID- 17908876 TI - Magnetic resonance imaging of soft-tissue tumors: determinate and indeterminate lesions. PMID- 17908877 TI - Anterior cruciate ligament reconstruction with use of autologous quadriceps tendon graft. PMID- 17908878 TI - Systematic review of arthroscopic rotator cuff repair and mini-open rotator cuff repair. PMID- 17908879 TI - Intraoperative assessment of bone cuts to guide surgical technique during total knee arthroplasty. PMID- 17908880 TI - Future clinical and economic impact of revision total hip and knee arthroplasty. PMID- 17908881 TI - Effects of electrical physical stimuli on articular cartilage. PMID- 17908882 TI - Stress over the anterior aspect of the knee with kneeling. PMID- 17908883 TI - The lateral intercondylar ridge--a key to anatomic anterior cruciate ligament reconstruction. PMID- 17908884 TI - A randomized trial comparing autologous chondrocyte implantation with microfracture. Findings at five years. AB - BACKGROUND: The optimal treatment for cartilage lesions has not yet been established. The objective of this randomized trial was to compare autologous chondrocyte implantation with microfracture. This paper represents an update, with presentation of the clinical results at five years. METHODS: Eighty patients who had a single chronic symptomatic cartilage defect on the femoral condyle in a stable knee without general osteoarthritis were included in the study. Forty patients were treated with autologous chondrocyte implantation, and forty were treated with microfracture. We used the International Cartilage Repair Society, Lysholm, Short Form-36, and Tegner forms to collect clinical data, and radiographs were evaluated with use of the Kellgren and Lawrence grading system. RESULTS: At two and five years, both groups had significant clinical improvement compared with the preoperative status. At the five-year follow-up interval, there were nine failures (23%) in both groups compared with two failures of the autologous chondrocyte implantation and one failure of the microfracture treatment at two years. Younger patients did better in both groups. We did not find a correlation between histological quality and clinical outcome. However, none of the patients with the best-quality cartilage (predominantly hyaline) at the two-year mark had a later failure. One-third of the patients in both groups had radiographic evidence of early osteoarthritis at five years. CONCLUSIONS: Both methods provided satisfactory results in 77% of the patients at five years. There was no significant difference in the clinical and radiographic results between the two treatment groups and no correlation between the histological findings and the clinical outcome. One-third of the patients had early radiographic signs of osteoarthritis five years after the surgery. Further long term follow-up is needed to determine if one method is better than the other and to study the progression of osteoarthritis. PMID- 17908885 TI - Reliability of histopathologic and radiologic grading of cartilaginous neoplasms in long bones. AB - BACKGROUND: Providing the best treatment options and appropriate prognostic information to patients with cartilaginous neoplasms of long bones depends on distinguishing benign from malignant lesions. Correlative interpretation of imaging, histopathology, and clinical information is the current method for making this distinction, yet the reliability of this approach has not been critically evaluated. This study quantifies the interobserver reliability of the determination of grade for cartilaginous neoplasms among a group of experienced musculoskeletal pathologists and radiologists. METHODS: Nine recognized musculoskeletal pathologists and eight recognized musculoskeletal radiologists reviewed forty-six consecutive cases of cartilaginous lesions in long bones that underwent open biopsy or intralesional curettage. All diagnosticians had a bulleted history and preoperative conventional radiographs for review. Pathologists reviewed the original hematoxylin and eosin-stained glass slides from each case. Radiologists reviewed any additional imaging that was available, variably including serial radiographs, magnetic resonance imaging, and computed tomography scans. Each diagnostician classified a lesion as benign, low-grade malignant, or high-grade malignant. Kappa coefficients were calculated as a measure of reliability. RESULTS: Kappa coefficients for interrater reliability were 0.443 for the pathologists and 0.345 for the radiologists (p < 0.0001 for both). Kappa coefficients for a subgroup of cases determined to be high risk by subsequent clinical course were poorer at 0.236 and 0.206, respectively (p < 0.0001 for both). Slightly improved agreement among radiologists was noted for the twenty lesions that had magnetic resonance imaging available (Kappa = 0.437, p < 0.0001), but not for the lesions analyzed with serial plain radiographs or computed tomography scans. CONCLUSIONS: This study demonstrates low reliability for the grading of cartilaginous lesions in long bones, even among specialized and experienced pathologists and radiologists. This included low reliability both in differentiating benign from malignant lesions and in differentiating high grade from low-grade malignant lesions, both of which are critical to the safe treatment of these neoplasms. This may explain in part the wide variation in outcomes reported for chondrosarcomas treated in different medical centers. New diagnostic and grading strategies linked to protocol-driven treatments are needed, but they must be measured against the long-term gold standard of patient outcomes. PMID- 17908886 TI - Immobilization in external rotation after shoulder dislocation reduces the risk of recurrence. A randomized controlled trial. AB - BACKGROUND: An initial anterior dislocation of the shoulder becomes recurrent in 66% to 94% of young patients after immobilization of the shoulder in internal rotation. Magnetic resonance imaging and studies of cadavera have shown that coaptation of the Bankart lesion is better with the arm in external rotation than it is with the arm in internal rotation. Our aim was to determine the benefit of immobilization in external rotation in a randomized controlled trial. METHODS: One hundred and ninety-eight patients with an initial anterior dislocation of the shoulder were randomly assigned to be treated with immobilization in either internal rotation (ninety-four shoulders) or external rotation (104 shoulders) for three weeks. The primary outcome measure was a recurrent dislocation or subluxation. The minimum follow-up period was two years. RESULTS: The follow-up rate was seventy-four (79%) of ninety-four in the internal rotation group and eighty-five (82%) of 104 in the external rotation group. The compliance rate was thirty-nine (53%) of seventy-four in the internal rotation group and sixty-one (72%) of eighty-five in the external rotation group (p = 0.013). The intention-to treat analysis revealed that the recurrence rate in the external rotation group (twenty-two of eighty-five; 26%) was significantly lower than that in the internal rotation group (thirty-one of seventy-four; 42%) (p = 0.033) with a relative risk reduction of 38.2%. In the subgroup of patients who were thirty years of age or younger, the relative risk reduction was 46.1%. CONCLUSIONS: Immobilization in external rotation after an initial shoulder dislocation reduces the risk of recurrence compared with that associated with the conventional method of immobilization in internal rotation. This treatment method appears to be particularly beneficial for patients who are thirty years of age or younger. PMID- 17908887 TI - The economic impact of reprocessing external fixation components. AB - BACKGROUND: The trend toward temporizing external fixation of complex fractures has resulted in increased expenditures for these devices. Increasing pressure to reduce health-care expenditures has led to exploration of reuse of equipment intended for single use. Devices must be tested and recertified prior to redeployment in hospital stock. We report the rate of manufacturer recertification and institutional cost savings associated with a reuse program approved by the United States Food and Drug Administration. METHODS: All Hoffmann II external fixation components that had been removed at our institution during the study period were submitted to the manufacturer for visual inspection and mechanical testing. Pass rates for original components and previously recycled components were determined. With use of a conservative pass rate and the assumption of a maximum of three recertifications of each component, the total potential hospital savings on external fixation were calculated. RESULTS: The first pass rate was 76%. The second pass rate (i.e., the rate for components that had already been recertified once and had been sent for a second recertification) was 83%, but that rate was derived from a limited sample. On the basis of a conservative pass-rate estimate of 75%, the predicted average number of uses of a recyclable component was 2.7. The recertified components were sold back to our hospital at 50% of the original price. Because carbon-fiber bars and half-pins are not recycled, 85% of the charges expended on a new external fixation component are spent on portions of the system that are recyclable. The potential total savings on reusable components was found to be 32%, with a total savings of 27% for the whole external fixation system. No recertified components failed in clinical use over the course of the study. CONCLUSIONS: With the expansion of cost-control efforts, the recycling of medical devices appears inevitable. Previous data have demonstrated the safety of reuse of external fixation devices, and this study confirms that finding. Our paper demonstrates the real cost savings associated with a manufacturer-based testing and recertification program. Issues of voluntary participation in reuse programs, component ownership, and the impact of savings on patient charges are yet to be worked out by individual institutions. PMID- 17908888 TI - Unicondylar osteoarticular allografts of the knee. AB - BACKGROUND: In the management of a resected distal femoral or proximal tibial condyle as the result of tumor or trauma, a unicondylar osteoarticular allograft is currently the only reconstructive option that avoids the sacrifice of the unaffected condyle. The purposes of this study were to perform a survival analysis of unicondylar osteoarticular allografts of the knee and to evaluate the complications. METHODS: We retrospectively reviewed the results of forty large unicondylar osteoarticular allograft procedures in thirty-eight patients who were followed for a mean of eleven years. Twenty-nine allografts were femoral transplants and included eleven medial and eighteen lateral femoral condyles. Eleven allografts were tibial transplants, including four medial and seven lateral tibial condyles. The procedure was performed after a tumor resection in thirty-six patients and to replace condylar loss after a severe open fracture in the remaining two patients. Complications were analyzed, and allograft survival from the date of implantation to the date of revision or the time of the latest follow-up was determined. Functional and radiographic results were documented according to the Musculoskeletal Tumor Society scoring system at the time of the latest follow-up. RESULTS: One patient died of tumor-related causes without allograft failure before the two-year follow-up evaluation. The global rate of allograft survival at both five and ten years was 85%, with a mean follow-up of 148 months. In six patients, the allografts were removed at an average of twenty six months (range, six to forty-eight months) and these were considered failures. All six patients underwent a second allograft procedure including two new unicondylar and four bicondylar reconstructions. The mean radiographic score for the thirty-three surviving allografts evaluated was 89%, with an average functional score of 27 of a possible 30 points. CONCLUSIONS: Unicondylar osteoarticular allografts of the knee appear to be a reliable alternative for patients in whom reconstruction of massive osteoarticular bone loss is limited to one condyle of the femur or the tibia. PMID- 17908889 TI - Comparison of reoperation rates following ankle arthrodesis and total ankle arthroplasty. AB - BACKGROUND: The role of ankle arthroplasty in the treatment of ankle arthritis is controversial. Ankle fusion is commonly performed, but there is ongoing concern about functional limitations and arthritis in the adjacent subtalar joint following ankle arthrodesis. The use of ankle arthroplasty as an alternative to ankle fusion is expanding, but reported results have been limited to those in case series. The purpose of this study was to compare the reoperation rates following ankle arthrodesis and ankle replacement on the basis of observational, population-based data from all inpatient admissions in California over a ten-year period. Our hypothesis was that patients treated with ankle replacement would have a lower risk of undergoing subtalar fusion but a higher overall risk of undergoing major revision surgery. METHODS: We used California's hospital discharge database to identify patients who had undergone ankle replacement or ankle arthrodesis as inpatients in the years 1995 through 2004. Short-term outcomes, including rates of major revision surgery, pulmonary embolism, amputation, and infection, were examined. Long-term outcomes that were analyzed included the rates of major revision surgery and subtalar joint fusion. Logistic and proportional hazard regression models were used to estimate the impact of the choice of ankle replacement or ankle fusion on the rates of adverse outcomes, with adjustment for patient factors including age and comorbidity. RESULTS: A total of 4705 ankle fusions and 480 ankle replacements were performed during the ten-year study period. Patients who had undergone ankle replacement had an increased risk of device-related infection and of having a major revision procedure. The rates of major revision surgery after ankle replacement were 9% at one year and 23% at five years compared with 5% and 11% following ankle arthrodesis. Patients treated with ankle arthrodesis had a higher rate of subtalar fusion at five years postoperatively (2.8%) than did those treated with ankle replacement (0.7%). Regression analysis confirmed a significant increase in the risk of major revision surgery (hazard ratio, 1.93 [95% confidence interval, 1.50 to 2.49]; p < 0.001) but a decreased risk of subtalar fusion (hazard ratio, 0.28 [95% confidence interval, 0.09 to 0.87]; p = 0.03) in patients treated with ankle replacement compared with those treated with ankle fusion. CONCLUSIONS: This study confirms that, compared with ankle fusion, ankle replacement is associated with a higher risk of complications but also potential advantages in terms of a decreased risk of the patient requiring subtalar joint fusion. Additional controlled trials are needed to clarify the appropriate indications for ankle arthrodesis and ankle replacement. PMID- 17908890 TI - Morphology of the femoral intercondylar notch. AB - BACKGROUND: During anterior cruciate ligament reconstruction, proper femoral tunnel placement is important. The purpose of the present study was to characterize the osseous anatomy of the femoral intercondylar notch. METHODS: We studied the morphology of the femoral intercondylar notch in 200 human femora from skeletally mature donors, with specific attention being paid to the morphology of the ridge on the lateral wall of the intercondylar notch and the posterolateral rim of the intercondylar notch. The distances from the posterolateral rim of the intercondylar notch to the lateral intercondylar ridge and from the posterolateral rim of the intercondylar notch to the inlet of the intercondylar notch (notch depth) were measured at the nine, ten, and eleven o'clock positions for right knees and at the one, two and three o'clock positions for left knees. RESULTS: The lateral intercondylar ridge was present in 194 femora and absent in six. The mean distance from the posterolateral rim of the intercondylar notch to the lateral intercondylar ridge was 9.0, 11.0, and 12.7 mm at the nine, ten, and eleven o'clock positions in right knees and the one, two, and three o'clock positions in left knees, respectively. We observed three different types of morphology of the posterolateral rim of the intercondylar notch. The morphology of the posterolateral rim of the intercondylar notch was distinct in 183 of 200 specimens. A distinct, straight border (type 1) was seen in 175 femora (87.5%); a distinct, V-shaped border (type 2) was seen in eight (4%); and an indistinct border (type 3) was seen in seventeen (8.5%). CONCLUSIONS: The morphology of the femoral intercondylar notch varies little. Occasionally, the posterolateral rim of the intercondylar notch is not well defined. In these knees, accurate placement of commercial femoral tunnel aiming guides may be difficult. PMID- 17908891 TI - Radial head excision and synovectomy in patients with hemophilia. AB - BACKGROUND: Chronic hemophilic synovitis of the elbow usually leads to enlargement and erosion of the radial head, resulting in mechanical blockage of forearm rotation, synovial impingement, recurrent hemarthrosis, and pain. The purpose of the present study was to evaluate the intermediate-term results of radial head excision and synovectomy in a large group of patients with hemophilia who had been managed at a single institution. METHODS: Information on forty radial head excision and synovectomy procedures that had been performed at our institution from 1969 to 2004 was retrospectively collected. All but one of the operations had been performed in patients with severe hemophilia. The mean age of the patients at the time of the procedure was thirty-three years. Pain, limited range of motion, and bleeding were the indications for surgery. The mean duration of follow-up was 7.7 years. RESULTS: Only one postoperative complication was observed: a posterior interosseous nerve palsy that fully resolved by six months. No additional surgical intervention for bleeding was required in sixteen of the nineteen elbows in which bleeding was one of the indications for surgery. Of the forty elbows, seven required a secondary surgical procedure at a mean of five years after the excision of the radial head. Examination of the mean range of motion at the time of the latest follow-up demonstrated a 63 degrees increase in the pronation-supination arc (p < 0.00001) but only a 2 degrees increase in the flexion arc. CONCLUSIONS: Radial head excision in patients with hemophilia is an effective procedure for improving forearm rotation and reducing pain and bleeding frequency, with a low risk of complications. PMID- 17908892 TI - The shape of the lateral edge of the first metatarsal head as a risk factor for recurrence of hallux valgus. AB - BACKGROUND: The relationship between the shape of the first metatarsal head and hallux valgus deformity remains controversial. The purpose of the present study was to retrospectively analyze differences in the radiographic appearance of the shape of the lateral edge of the first metatarsal head between women with normal feet and those with hallux valgus and to clarify the relationship between the shape of the lateral edge and the postoperative recurrence of hallux valgus deformity. METHODS: Dorsoplantar weight-bearing radiographs of sixty normal feet in women (the control group) and sixty feet in women with hallux valgus (the hallux valgus group) were reviewed. The feet in the hallux valgus group were treated with a proximal metatarsal osteotomy, and the radiographs of those feet were assessed preoperatively, at the time of early follow-up (mean, 3.4 months), and at the time of the most recent follow-up (mean, forty-eight months). The shape of the lateral edge, which was defined as consisting of the articular and lateral surfaces of the first metatarsal head, was examined. The shape of the lateral edge was classified as one of three types: round (type R), angular (type A), and intermediate (type I). We defined the round sign as being positive when the shape of the lateral edge was classified as type R. RESULTS: Prior to surgery, the prevalence of the type-R shape was significantly greater in the hallux valgus group than it was in the control group (78.3% compared with 1.7%; p < 0.0001) and the prevalence of type-A shape was significantly lower in the hallux valgus group than in the control group (3.3% compared with 81.7%; p < 0.0001). In the hallux valgus group, the prevalence of the type-R shape at the time of the early follow-up after surgery was significantly lower than that before surgery (p < 0.0001). Feet with a positive round sign at the time of the early follow-up had a greater risk of having recurrence of the hallux valgus deformity at the time of the most recent follow-up than did those without a round sign at the time of the early follow-up (odds ratio, 12.71; 95% confidence interval, 3.21 to 50.36). CONCLUSIONS: There is a significant relationship between a round-shaped lateral edge of the first metatarsal head and hallux valgus, and a positive round sign after a proximal first metatarsal osteotomy can be a risk factor for the recurrence of hallux valgus. PMID- 17908893 TI - Adductor-related groin pain in competitive athletes. Role of adductor enthesis, magnetic resonance imaging, and entheseal pubic cleft injections. AB - BACKGROUND: Adductor dysfunction is a condition that can cause groin pain in competitive athletes, but the source of the pain has not been established and no specific interventions have been evaluated. We previously defined a magnetic resonance imaging protocol to visualize adductor enthesopathy. The aim of this study was to elucidate, in the context of adductor-related groin pain in the competitive athlete, the role of the adductor enthesis (origin), the relevance of adductor enthesopathy diagnosed with magnetic resonance imaging, and the efficacy of entheseal pubic cleft injections of local anesthetic and steroids. METHODS: We reviewed the findings in a consecutive series of twenty-four competitive athletes who had presented to our sports medicine clinic with groin pain secondary to adductor longus dysfunction. Magnetic resonance imaging was performed to assess the adductor longus origin for the presence or absence of enthesopathy. Seven patients (Group 1) had no evidence of enthesopathy on magnetic resonance imaging, and seventeen patients (Group 2) had enthesopathy confirmed on magnetic resonance imaging. All patients were treated with a single pubic cleft injection of local anesthetic and steroid into the adductor enthesis. At one year after this treatment, the patients were assessed for recurrence of symptoms. RESULTS: On clinical reassessment five minutes after the injection, all twenty-four athletes reported resolution of the groin pain. At one year, none of the seven patients in Group 1 had experienced a recurrence. Sixteen of the seventeen patients in Group 2 had a recurrence of the symptoms (p < 0.001) at a mean of five weeks (range, one to sixteen weeks) after the injection. CONCLUSIONS: A single entheseal pubic cleft injection can be expected to afford at least one year of relief of adductor related groin pain in a competitive athlete with normal findings on a magnetic resonance imaging scan; however, it should be employed only as a diagnostic test or short-term treatment for a competitive athlete with evidence of enthesopathy on magnetic resonance imaging. PMID- 17908894 TI - Magnetic resonance imaging-guided percutaneous biopsy of musculoskeletal lesions. AB - BACKGROUND: Bone, soft-tissue, and articular lesions are often well visualized by magnetic resonance imaging. Our goal was to evaluate the diagnostic performance of magnetic resonance imaging-guided biopsies of selected musculoskeletal lesions. METHODS: In this retrospective case series, forty-five consecutive biopsies were performed in an open mid-field 0.5-T interventional magnetic resonance imaging unit with a real-time guidance system. The biopsies were performed at twenty bone, eighteen extra-articular soft-tissue, and seven intra articular soft-tissue sites. The main reasons for using magnetic resonance imaging guidance were the need to improve lesion conspicuity compared with that provided by other imaging modalities, the need for site-specific targeting within the lesion, and the need for real-time guidance. Samples were obtained with fine needle aspiration, core-needle biopsy, or a combination of these techniques. An independent reference standard was used to confirm the final diagnosis. Diagnostic performance was evaluated on the basis of the diagnostic yield (the proportion of biopsies yielding sufficient material for pathological evaluation) and diagnostic accuracy (sensitivity, specificity, positive predictive value, and negative predictive value). Complications were identified as well. RESULTS: The diagnostic yield was 91% (forty-one of forty-five biopsies yielded sufficient material for a diagnosis) overall, 95% (nineteen of twenty) for the bone lesions, 94% (seventeen of eighteen) for the extra-articular soft-tissue lesions, and 71% (five of seven) for the intra-articular soft-tissue lesions. With regard to the diagnostic accuracy, the sensitivity was 0.86, the specificity was 1.00, the positive predictive value was 1.00, and the negative predictive value was 0.76 in the overall group. The respective values were 0.92, 1.00, 1.00, and 0.86 for the bone lesions; 0.77, 1.00, 1.00, and 0.57 for the extra-articular soft-tissue lesions; and 1.00, 1.00, 1.00, and 1.00 for the intra-articular soft-tissue lesions. There was one complication: exacerbation of neuropathic pain related to a biopsy of a peripheral nerve sheath tumor. CONCLUSIONS: Magnetic resonance imaging-guided percutaneous biopsies of musculoskeletal lesions for which other imaging modalities might be inadequate have a good diagnostic performance overall. The performance can be very good for bone lesions, moderate for extra articular soft-tissue lesions, and fair for intra-articular soft-tissue lesions. PMID- 17908895 TI - Osteochondral transplantation to treat osteochondral lesions in the elbow. AB - BACKGROUND: Effective treatment of osteochondral lesions in the elbow remains challenging. Arthroscopic debridement and microfracture or retrograde drilling techniques are often insufficient and provide only temporary symptomatic relief. The purpose of this study was to evaluate the treatment of these lesions with osteochondral autografts. METHODS: From 1999 to 2002, seven patients with osteochondral lesions of the capitellum humeri (five patients), trochlea (one patient), or radial head (one patient) were treated with cylindrical osteochondral grafts, which were harvested from the non-weight-bearing area of the proximal aspect of the lateral femoral condyle. The patients (three female and four male patients with an average age of seventeen years) were evaluated preoperatively and postoperatively, with an average follow-up of fifty-nine months. The Broberg and Morrey score was chosen for functional evaluation of the elbow (with regard to motion, pain, strength, activities of daily living, and stability), and the American Shoulder and Elbow Surgeons score was used for the analysis of pain. All patients had imaging studies done preoperatively to evaluate the defect and postoperatively to assess the ingrowth and viability of the graft. The ipsilateral knee was examined for donor-site morbidity. RESULTS: The Broberg and Morrey score improved from a mean (and standard deviation) of 76.3 +/- 13.2 preoperatively to 97.6 +/- 2.7 postoperatively, and pain scores were significantly reduced (p < 0.05). The mean elbow extension lag of 4.7 degrees +/- 5.8 degrees was reduced to 0 degrees postoperatively. Compared with the contralateral side, there was a mean preoperative flexion lag of 12.9 degrees +/- 13.8 degrees . At the time of the final follow-up, flexion was free and was equal bilaterally in all patients. None of the plain radiographs made at the time of follow-up showed any degenerative changes or signs of osteoarthritis. The postoperative magnetic resonance imaging scans showed graft viability and a congruent chondral surface in all seven patients. No donor-site morbidity was noted at one year postoperatively. CONCLUSIONS: The osteochondral autograft procedure described in the present study provides the opportunity to retain viable hyaline cartilage for the repair of osteochondral lesions in the elbow while restoring joint congruity and function and perhaps reducing the risk of osteoarthritis. These medium-term results suggest that the risks of a two-joint procedure are modest and justifiable. In addition, the described technique provides an option for revision surgery after the failure of other surgical procedures. PMID- 17908896 TI - The effect of stuffing the patellofemoral compartment on the outcome of total knee arthroplasty. AB - BACKGROUND: The effect of so-called stuffing of the patellofemoral compartment at the time of total knee arthroplasty (that is, increasing the anterior patellar displacement, the anteroposterior femoral size, or the combined anteroposterior patellofemoral size) has not been well studied. The purpose of the present study was to evaluate the effect of stuffing the patellofemoral compartment on the outcome of primary total knee arthroplasty. METHODS: A retrospective review of 1100 primary total knee arthroplasties that had been performed in 1997 and 1998 was conducted. Eight hundred and thirty arthroplasties (75.5%) met the diagnostic and minimum two-year follow-up criteria for inclusion in this report. Radiographic measurements were made to determine preoperative and postoperative anterior patellar displacement, anteroposterior femoral size, combined anteroposterior patellofemoral size, anterior femoral offset, and posterior femoral offset. Regression analysis was performed to determine the effects of changes in these variables on the range of motion, the Knee Society Knee Score, the Knee Society Function Score, the Knee Society Pain Score, and the rate of lateral retinacular release. RESULTS: Preoperative to postoperative changes in anterior patellar displacement, anteroposterior femoral size, combined anteroposterior patellofemoral size, anterior femoral offset, and posterior femoral offset had no clinically meaningful effect on the range of motion of the knee or on any of the Knee Society scores. Increases in anterior patellar displacement were associated with a lower probability of the need for a lateral retinacular release. Increases in measured anteroposterior femoral size were associated with a higher probability of the need for lateral release. Even when combined, however, these relationships explained only 10.1% of the observed variance in the need for lateral retinacular release. Moreover, analyses indicated that patient gender, large as opposed to medium patellar size, and absolute femoral component size influenced the likelihood of lateral release more than did anterior patellar displacement and measured anteroposterior femoral size. CONCLUSIONS: Our findings do not support the widely held belief that stuffing of the patellofemoral joint results in adverse outcomes after total knee arthroplasty. Furthermore, the need for lateral release appears to be multifactorial and likely involves a more complex set of factors. Thus, without evidence of other identifiable causes of failure, we do not recommend revision for the treatment of pain of an overstuffed knee joint. PMID- 17908897 TI - Hydroxyapatite-coated tibial implants compared with cemented tibial fixation in primary total knee arthroplasty. A randomized trial of outcomes at five years. AB - BACKGROUND: Although excellent long-term results have been reported with cemented tibial fixation, cementless fixation as a means to improve the longevity of total knee prostheses continues to be of interest to clinicians. The purpose of this study was to compare outcomes between cementless tibial fixation with hydroxyapatite and cemented tibial fixation in the first five years following primary total knee arthroplasty. METHODS: We performed a prospective, randomized clinical trial that included eighty-one patients with noninflammatory knee arthritis who underwent primary total knee arthroplasty when they were less than seventy years of age. The subjects were randomized at the time of surgery to be treated with either cementless tibial fixation with hydroxyapatite or cemented tibial fixation. Evaluations were performed preoperatively and at six months, one year, and five years postoperatively by a physical therapist who was blinded to group allocation. Self-reported pain and function, the primary outcomes, were measured with the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and the RAND 36-Item Health Survey (RAND-36). Complications and revision rates were determined through a review of hospital records and at each patient evaluation. The Knee Society radiographic score was used to evaluate plain radiographs at each assessment. RESULTS: Seventy subjects (86%) completed the five-year assessment. Slightly more pain was reported in the hydroxyapatite group at six months as measured with both the WOMAC and the RAND-36, a difference that disappeared by one year postoperatively. No differences were seen in function, radiographic findings, or complications. No subject required revision of the tibial prosthesis during the study. CONCLUSIONS: At five years postoperatively, there is no difference between cementless tibial fixation with hydroxyapatite and cemented tibial fixation in terms of self-reported pain, function, health-related quality of life, postoperative complications, or radiographic scores. PMID- 17908898 TI - Rim cracking of the cross-linked longevity polyethylene acetabular liner after total hip arthroplasty. AB - BACKGROUND: Studies have suggested that cross-linked polyethylene bearings reduce wear rates from 40% to 100% compared with conventional polyethylene. However, the reduced mechanical properties of highly cross-linked polyethylene have the potential to be a limiting factor in device performance. We reviewed a series of retrieved acetabular liners with a fracture of the superior rim to assess the factors that played a role in their failure. METHODS: Four Longevity acetabular bearings, which had been retrieved from two patients after seven to twenty-seven months in vivo, were visually examined for clinical damage, were assessed with use of Fourier transform infrared spectroscopy to determine the level of oxidation, and were analyzed for mechanical properties and fracture surface characterization. Control data were obtained from never-implanted devices and from global reference ultrahigh molecular weight polyethylene bar stock as an industry calibration material. RESULTS: All four retrieved liners demonstrated articular surface wear modes, which in most cases were rated as moderate, and none were rated as severe. All showed cracking or rim failure of the liner at the superior aspect along the groove in the polyethylene that engages the locking ring of the shell. The retrieved liners had no measurable oxidation, and the mechanical properties were comparable with those of never-implanted material. CONCLUSIONS: There was no notable in vivo degradation of the retrieved liners. Important factors related to failure appear to be thin polyethylene at the cup rim, relatively vertical cup alignment, and the material properties of the highly cross-linked polyethylene that are decreased relative to conventional polyethylene. The critical dimension with respect to rim failure in modular liners appears to be the minimum thickness at the equatorial region. PMID- 17908899 TI - Distal tibial reconstruction with use of a circular external fixator and an intramedullary nail. The combined technique. AB - BACKGROUND: Distal tibial reconstruction with use of an external fixator when there is bone loss, limb-length discrepancy, and/or ankle instability is associated with many problems. The technique of limb-lengthening, ankle arthrodesis, and segmental transfer over an intramedullary nail has been introduced to overcome these problems. The present study investigates this combined technique. METHODS: Between 2002 and 2005, six patients, who ranged from seventeen to seventy years old, underwent distal tibial reconstruction and ankle arthrodesis with use of a circular external fixator and an intramedullary nail to treat a distal tibial defect following resection for chronic osteomyelitis or tumor or to treat a limb-length discrepancy combined with ankle instability. Functional and radiographic results were evaluated, with use of the criteria described by Paley et al., at an average follow-up of thirty-four months. RESULTS: The mean size of the bone defects in three patients was 5.3 cm (2, 7, and 7 cm), and the mean amount of the limb-shortening in four patients was 5.25 cm (range, 4 to 6 cm). The mean external fixation time was 3.5 months, and the mean external fixator index was 0.57 mo/cm. There was no recurrence of infection in the two patients with osteomyelitis. All six patients had excellent bone results, and the functional results were excellent for two patients and good for four patients. There were four complications, three of which were categorized, according to Paley, as a problem (a difficulty that occurs during lengthening and is resolved without operative intervention) and one that was categorized as an obstacle (a difficulty that occurs during lengthening and needs operative treatment). CONCLUSIONS: The combined technique is an improvement over the classic external fixation techniques of distal tibial reconstruction with ankle arthrodesis. It reduces the duration of external fixation, thus increasing patient acceptance, and it is associated with a low complication rate facilitating more rapid rehabilitation. PMID- 17908900 TI - Lisfranc joint displacement following sequential ligament sectioning. AB - BACKGROUND: There are two primary radiographic patterns of Lisfranc instability, transverse and longitudinal. There is no single diagnostic method with which to consistently confirm the diagnosis of an unstable injury. Our purpose was to define which ligament disruptions produce these two injury patterns and to compare the utility of weight-bearing and stress radiographs for detecting each pattern of instability. METHODS: Ten fresh-frozen cadaveric lower extremities were dissected to expose the dorsal aspect of the midfoot. Radiographic markers were placed at the base of the second metatarsal and the distal borders of the first and second cuneiforms. The specimens underwent sectioning of the interosseous first cuneiform-second metatarsal (Lisfranc) ligament and were then divided into two groups. The transverse group underwent sectioning of the plantar ligament between the first cuneiform and the second and third metatarsals at the plantar aspect of the second cuneiform-second metatarsal joint, whereas the longitudinal group underwent sectioning of the interosseous ligament between the first and second cuneiforms. Weight-bearing, adduction, and abduction stress radiographs were made before and after each ligament was sectioned. The radiographs were digitized, and displacement was recorded. Instability was defined as >or=2 mm of displacement. RESULTS: Weight-bearing radiographs made after the Lisfranc (first cuneiform-second metatarsal) ligament alone was sectioned were diagnostic (showed instability) for one of ten specimens. Abduction stress radiographs were diagnostic for two of five specimens, and adduction stress radiographs were diagnostic for zero of five specimens. In the transverse group (sectioning of the plantar ligament between the first cuneiform and the second and third metatarsals), weight-bearing radiographs were diagnostic on the basis of first cuneiform-second metatarsal displacement for one of five specimens but were not diagnostic on the basis of second cuneiform-second metatarsal displacement for any of five specimens. Abduction stress radiographs were diagnostic on the basis of displacement of both the first cuneiform-second metatarsal and the second cuneiform-second metatarsal joints for five of five specimens. In the longitudinal group (sectioning of the interosseous ligament between the first and second cuneiforms), weight-bearing radiographs were diagnostic on the basis of first cuneiform-second metatarsal displacement for one of five specimens and were diagnostic on the basis of displacement between the first and second cuneiforms for one of five specimens. Adduction stress radiographs were diagnostic on the basis of first cuneiform-second metatarsal displacement for one of five specimens and were diagnostic on the basis of displacement between the first and second cuneiforms for four of five specimens. CONCLUSIONS: Transverse instability required sectioning of both the interosseous first cuneiform-second metatarsal ligament and the plantar ligament between the first cuneiform and the second and third metatarsals. Longitudinal instability required sectioning of both the interosseous first cuneiform-second metatarsal ligament and the interosseous ligament between the first and second cuneiforms. Compared with weight-bearing radiographs, injury-specific manual stress radiographs showed qualitatively greater displacement when used to evaluate both patterns of instability. PMID- 17908901 TI - Reconstruction of large skeletal defects due to osteomyelitis with the vascularized fibular graft in children. AB - BACKGROUND: Reconstruction of large skeletal defects secondary to osteomyelitis is a challenging problem. The purpose of this study was to evaluate the outcome of the use of a vascularized fibular graft to treat such defects in children. METHODS: Eight patients with a mean age of seven years and a skeletal defect with a mean length of 11.8 cm (range, 6 to 17 cm) were treated with a vascularized fibular graft. A staged protocol was used for the five patients with an active infection at the time of presentation. The first procedure consisted of radical debridement, and at the second stage a free (seven patients) or pedicled (one patient) vascularized fibular graft was used. The mean follow-up time was 5.7 years. RESULTS: Union of the graft occurred primarily in seven of the eight patients, at a mean of 3.5 months, and after iliac crest bone-grafting in the remaining patient. There was no recurrence of deep infection. Complications developed in two patients. The mean time to full weight-bearing by the seven patients with a lower-extremity reconstruction was 8.4 months, and all patients were pain-free and able to walk without supportive devices. CONCLUSIONS: A vascularized fibular graft is a viable option for the management of large skeletal defects resulting from osteomyelitis in children. PMID- 17908902 TI - Bioprotection of tendon repair: adjunctive use of botulinum toxin A in Achilles tendon repair in the rat. AB - BACKGROUND: Tendon-repair techniques have evolved to increase the construct strength of the repair site in order to permit early active range of motion without tendon gap or rupture. The present study evaluated the hypothesis that the injection of botulinum neurotoxin type-A (BoNT-A) into the gastrocnemius muscle will reduce the active force production of that muscle below the force required to rupture the associated, repaired Achilles tendon. METHODS: Seventy nine rat Achilles tendons were surgically bisected and were repaired with use of a two-strand core suture with a running epitenon repair. After the repair, the animals were treated with unilateral intramuscular (gastrocnemius) injections of either BoNT-A (6 U/kg body weight) (thirty-seven rats) or saline solution (forty two rats). Operatively treated ankles were fixed in the neutral position with a percutaneous pin for the first two days after surgery. Unrestricted ankle motion and weight-bearing were allowed after the second postoperative day. An assessment of gap formation or rupture at the repair site, electrophysiologic measurements of force applied to the tendon, and an assessment of the strength of the repaired tendon were performed. RESULTS: Intramuscular BoNT-A injections produced a significant, reversible reduction in active muscle force (p < 0.007). Twitch and tetanus contractions decreased to approximately 25% of the values for the control side within one week, remained at <50% of the values for the control side at one month, and returned to normal levels by six months. The tetanic force capability of the muscles that had been injected with BoNT-A was fivefold to tenfold less than the force required to rupture the associated Achilles tendon for as long as four weeks after tendon repair. The spontaneous Achilles tendon rupture rate of repaired tendons in the BoNT-A group was three times lower than that in the saline solution group at one week, and the tendon rupture force was significantly higher in the BoNT-A group between one and three weeks after repair (p < 0.007). There was no significant difference in tendon rupture force between the two groups after three weeks. CONCLUSIONS: Intramuscular gastrocnemius BoNT-A injections were associated with a significant reduction in force-generating potential, such that the muscle was incapable of actively producing enough force to rupture the repaired Achilles tendon in this rat model of tendon repair. PMID- 17908904 TI - Clavicular anatomy and the applicability of precontoured plates. AB - BACKGROUND: Plate fixation of clavicular fractures is technically difficult because of the complex anatomy of the bone, with an S-shaped curvature and a cephalad-to-caudad bow. The purpose of the present study was to characterize variations in clavicular anatomy and to determine the clinical applicability of an anatomic precontoured clavicular plate designed for fracture fixation. METHODS: One hundred pairs of clavicles were analyzed. The location and magnitude of the superior clavicular bow were determined with use of a digitizer and modeling software. Axial radiographs were made of each clavicle and the precontoured Acumed Locking Clavicle Plate, which is designed to be applied superiorly. With use of Adobe Photoshop technology, the plates were freely translated and rotated along each clavicle to determine the quality of fit and the location of the "best fit." RESULTS: The location of the maximum superior bow was lateral, with a mean distance of 37.2 +/- 18.4 mm from the acromial articulation and with a mean magnitude of 5.1 +/- 5.9 mm. There was no significant difference in the location or magnitude of the apex of the bow between specimens from male and female donors. The anatomic precontoured clavicular plate had the best fit in specimens from black male donors and the worst fit in specimens from white female donors, with a poor fit being seen in 38% (nineteen) of the fifty specimensfrom white female donors. The best location for superior plate application was along the medial aspect of the clavicle. CONCLUSIONS: The apex of the superior bow of the clavicle is typically located along the lateral aspect of the bone, whereas the medial aspect of the superior surface of the clavicle remains relatively flat, making it an ideal plating surface. The precontoured anatomic clavicular plate appears to fit the S-shaped curvature on the superior surface of the majority of clavicles in male patients but may not be as conforming in white female patients. While this plate fits in the medial three-fifths of the clavicle, it does not fit as well laterally. PMID- 17908903 TI - The effect of osteoclastic activity on tendon-to-bone healing: an experimental study in rabbits. AB - BACKGROUND: Healing of a tendon graft in a bone tunnel depends on bone ingrowth into the interface between tendon and bone. Excessive osteoclastic activity may contribute to bone resorption, tunnel widening, and impaired healing. We hypothesized that inhibition of osteoclastic activity by osteoprotegerin (OPG) would increase bone formation around a tendon graft in anterior cruciate ligament reconstruction in a rabbit model, while increased osteoclastic activity due to the application of receptor activator of nuclear factor-kappa B ligand (RANKL) would impair bone ingrowth. METHODS: Sixty skeletally mature, male New Zealand White rabbits underwent bilateral anterior cruciate ligament reconstruction. OPG (100 microg per tunnel) or RANKL (10 microg per tunnel) was delivered to the tendon-bone interface with use of a synthetic calcium phosphate carrier vehicle. Twenty animals were killed at two, four, and eight weeks after surgery. Two rabbits from each group were prepared for histological evaluation, and the other rabbits were used for biomechanical testing. RESULTS: A significantly greater amount of bone surrounded the tendon at the healing tendon-bone interface in the OPG-treated limbs compared with the controls and the RANKL-treated limbs at all time-points (p < 0.05). There were significantly fewer osteoclasts in the OPG treated limbs compared with the controls and the RANKL-treated limbs (p < 0.05). The average tunnel area in the OPG group was significantly smaller than that in the RANKL group (p = 0.003 at two weeks and p = 0.004 at four weeks). The femur anterior cruciate ligament-tibia complex of the OPG-treated limbs had significantly increased stiffness compared with RANKL-treated limbs at eight weeks (p = 0.04). CONCLUSIONS: Osteoprotegerin significantly improves bone formation around the grafted tendon and improves the stiffness at the healing tendon-bone junction in a rabbit model. PMID- 17908905 TI - Aspherical femoral head with highly cross-linked ultra-high molecular weight polyethylene surface cracking. A case report. PMID- 17908906 TI - Failure at the taper lock of a modular stemmed femoral implant in revision knee arthroplasty. A report of two cases and a retrieval analysis. PMID- 17908907 TI - Intracranial hemorrhage following incidental durotomy during spinal surgery. A report of four patients. PMID- 17908909 TI - Periprosthetic patellar fractures. PMID- 17908908 TI - Fractures associated with computer-navigated total knee arthroplasty. A report of two cases. PMID- 17908910 TI - Musculoskeletal medicine educational reform in the bone and joint decade. PMID- 17908911 TI - AOA-JOA Traveling Fellowship 2006: The Fellows' Travel Log. PMID- 17908912 TI - What's new in orthopaedic rehabilitation. PMID- 17908913 TI - Two electrical potential-dependent steps are required for transport by the Escherichia coli Tat machinery. AB - The twin-arginine translocation (Tat) pathway in Escherichia coli transports fully folded and assembled proteins across the energy-transducing periplasmic membrane. In chloroplasts, Tat transport requires energy input only from the proton motive force. To elucidate the mechanism and energetics of bacterial Tat protein transport, we developed an efficient in vitro transport assay using TatABC-enriched inverted membrane vesicles and the physiological precursor pre SufI. We report transport efficiencies of 60-80% for nanomolar pre-SufI concentrations. Dissipation of the pH gradient does not reduce pre-SufI transport efficiency. Instead, pre-SufI transport requires at least two electrical potential (Deltapsi)-dependent steps that differ in both the duration and minimum magnitude of the required Deltapsi. The data are consistent with a model in which a substantial Deltapsi of short duration is required for an early transport step, and in which a small Deltapsi of long duration is necessary to drive a later transport step. PMID- 17908914 TI - JAK1-STAT1-STAT3, a key pathway promoting proliferation and preventing premature differentiation of myoblasts. AB - Skeletal muscle stem cell-derived myoblasts are mainly responsible for postnatal muscle growth and injury-induced muscle regeneration. However, the cellular signaling pathways controlling the proliferation and differentiation of myoblasts are not fully understood. We demonstrate that Janus kinase 1 (JAK1) is required for myoblast proliferation and that it also functions as a checkpoint to prevent myoblasts from premature differentiation. Deliberate knockdown of JAK1 in both primary and immortalized myoblasts induces precocious myogenic differentiation with a concomitant reduction in cell proliferation. This is caused, in part, by an accelerated induction of MyoD, myocyte enhancer-binding factor 2 (MEF2), p21Cip1, and p27Kip1, a faster down-regulation of Id1, and an increase in MEF2 dependent gene transcription. Downstream of JAK1, of all the signal transducer and activator of transcriptions (STATs) present in myoblasts, we find that only STAT1 knockdown promotes myogenic differentiation in both primary and immortalized myoblasts. Leukemia inhibitory factor stimulates myoblast proliferation and represses differentiation via JAK1-STAT1-STAT3. Thus, JAK1 STAT1-STAT3 constitutes a signaling pathway that promotes myoblast proliferation and prevents premature myoblast differentiation. PMID- 17908915 TI - SMK-1/PPH-4.1-mediated silencing of the CHK-1 response to DNA damage in early C. elegans embryos. AB - During early embryogenesis in Caenorhabditis elegans, the ATL-1-CHK-1 (ataxia telangiectasia mutated and Rad3 related-Chk1) checkpoint controls the timing of cell division in the future germ line, or P lineage, of the animal. Activation of the CHK-1 pathway by its canonical stimulus DNA damage is actively suppressed in early embryos so that P lineage cell divisions may occur on schedule. We recently found that the rad-2 mutation alleviates this checkpoint silent DNA damage response and, by doing so, causes damage-dependent delays in early embryonic cell cycle progression and subsequent lethality. In this study, we report that mutations in the smk-1 gene cause the rad-2 phenotype. SMK-1 is a regulatory subunit of the PPH-4.1 (protein phosphatase 4) protein phosphatase, and we show that SMK-1 recruits PPH-4.1 to replicating chromatin, where it silences the CHK-1 response to DNA damage. These results identify the SMK-1-PPH-4.1 complex as a critical regulator of the CHK-1 pathway in a developmentally relevant context. PMID- 17908916 TI - p66Shc mediates anoikis through RhoA. AB - Detachment of parenchymal cells from a solid matrix switches contextual cues from survival to death during anoikis. Marked shape changes accompany detachment and are thought to trigger cell death, although a working model to explain the coordination of attachment sensation, shape change, and cell fate is elusive. The constitutive form of the adapter Shc, p52Shc, confers survival properties, whereas the longer p66Shc signals death through association with cytochrome c. We find that cells that lack p66Shc display poorly formed focal adhesions and escape anoikis. However, reexpression of p66Shc restores anoikis through a mechanism requiring focal adhesion targeting and RhoA activation but not an intact cytochrome c-binding motif. This pathway stimulates the formation of focal adhesions and stress fibers in attached cells and tension-dependent cell death upon detachment. p66Shc may thus report attachment status to the cell by imposing a tension test across candidate anchorage points, with load failure indicating detachment. PMID- 17908917 TI - Stress-dependent relocalization of translationally primed mRNPs to cytoplasmic granules that are kinetically and spatially distinct from P-bodies. AB - Cytoplasmic RNA granules serve key functions in the control of messenger RNA (mRNA) fate in eukaryotic cells. For instance, in yeast, severe stress induces mRNA relocalization to sites of degradation or storage called processing bodies (P-bodies). In this study, we show that the translation repression associated with glucose starvation causes the key translational mediators of mRNA recognition, eIF4E, eIF4G, and Pab1p, to resediment away from ribosomal fractions. These mediators then accumulate in P-bodies and in previously unrecognized cytoplasmic bodies, which we define as EGP-bodies. Our kinetic studies highlight the fundamental difference between EGP- and P-bodies and reflect the complex dynamics surrounding reconfiguration of the mRNA pool under stress conditions. An absence of key mRNA decay factors from EGP-bodies points toward an mRNA storage function for these bodies. Overall, this study highlights new potential control points in both the regulation of mRNA fate and the global control of translation initiation. PMID- 17908918 TI - Heterotrimeric G protein signaling functions with dynein to promote spindle positioning in C. elegans. AB - Proper orientation and positioning of the mitotic spindle is essential for the correct segregation of fate determinants during asymmetric cell division. Although heterotrimeric G proteins and their regulators are essential for spindle positioning in many cell types, their mechanism of action remains unclear. In this study, we show that dyrb-1, which encodes a dynein light chain, provides a functional link between heterotrimeric G protein signaling and dynein activity during spindle positioning in Caenorhabditis elegans. Embryos depleted of dyrb-1 display phenotypes similar to a weak loss of function of dynein activity, indicating that DYRB-1 is a positive regulator of dynein. We find that the depletion of dyrb-1 enhances the spindle positioning defect of weak loss of function alleles of two regulators of G protein signaling, LIN-5 and GPR-1/2, and that DYRB-1 physically associates with these two proteins. These results indicate that dynein activity functions with regulators of G protein signaling to regulate common downstream effectors during spindle positioning in the early C. elegans embryo. PMID- 17908920 TI - Hepatic UDP-glucuronosyltransferases responsible for glucuronidation of thyroxine in humans. AB - To clarify the UDP-glucuronosyltransferase (UGT) isoform(s) responsible for the glucuronidation of the thyroid hormone thyroxine (T(4)) in the human liver, the T(4) glucuronidation activities of recombinant human UGT isoforms and microsomes from seven individual human livers were comparatively examined. Among the 12 recombinant human UGT1A and UGT2B subfamily enzymes examined, UGT1A1, UGT1A3, UGT1A9, and UGT1A10 showed definite activities for T(4) glucuronidation. These UGT1A enzymes, with the exception of UGT1A10, were detected in all of the human liver microsomes examined. Interindividual differences in T(4) glucuronidation activity were observed among the microsomes from the seven individual human livers, and the T(4) glucuronidation activity was closely correlated with beta estradiol 3-glucuronidation activity. Furthermore, Spearman correlation analysis for a relationship between the T(4) glucuronidation activity and the level of UGT1A1, UGT1A3, and UGT1A9 in the microsomes revealed that levels of UGT1A1 and UGT1A3, but not that of UGT1A9, were closely correlated with T(4) glucuronidation activity. T(4) glucuronidation activity in human liver microsomes was strongly inhibited by 26,26,26,27,27,27-hexafluoro-1alpha,23(S),25-trihydroxyvitamin D(3) (an inhibitor of UGT1A3), moderately inhibited by either bilirubin (an inhibitor of UGT1A1) or beta-estradiol (an inhibitor of UGT1A1 and UGT1A9), but not inhibited by propofol (an inhibitor of UGT1A9). These findings indicated strongly that glucuronidation of T(4) in the human liver was mediated by UGT1A subfamily enzymes, especially UGT1Al and UGT1A3, and further suggested that the interindividual differences would come from differences in the expression levels of UGT1A1 and UGT1A3 in individual human livers. PMID- 17908921 TI - Regulation of CYP2A6 protein expression by skatole, indole, and testicular steroids in primary cultured pig hepatocytes. AB - CYP2A6 is one of the enzymes involved in the hepatic metabolism of a naturally produced compound, skatole, in the pig. Low CYP2A6 activity has been linked to excessive accumulation of skatole in pig adipose tissue and development of the phenomenon "boar taint." CYP2A6 activity varies between male and female animals, suggesting the involvement of sex hormones in regulation of the enzyme activity and/or expression. The present study investigated whether pig hepatic CYP2A6 protein expression is regulated by the testicular steroids testosterone, androstenone, or estrone sulfate using primary cultured hepatocytes as a model system. The study has also examined whether CYP2A6 expression can be modulated by the boar taint compounds skatole and indole. The research has established that androstenone inhibits CYP2A6 protein expression at the concentration of 1, 10, and 100 nM by 55, 37, and 44%, respectively. In contrast to androstenone, skatole and indole (final concentrations, 1, 10, and 100 nM) had a stimulatory effect on CYP2A6 expression. The effect of indole was more pronounced than that of skatole (maximum induction by 145 and 70%, respectively). Estrone sulfate and testosterone did not have a significant effect on CYP2A6 protein level. This is, as far as we know, the first communication to report the regulation of pig hepatic CYP2A6 expression by steroids and boar taint compounds. The hormonal modulation of CYP2A6 expression might contribute to gender-related differences in pig hepatic CYP2A6 activity and skatole accumulation in pig adipose tissue. PMID- 17908922 TI - Glucuronidation of mycophenolic acid by Wistar and Mrp2-deficient TR- rat liver microsomes. AB - In humans, mycophenolic acid (MPA) is metabolized primarily by glucuronidation in the liver to mycophenolate ether glucuronide (MPAGe) and mycophenolate acyl glucuronide (MPAGa). We have previously reported that in perfused livers of TR(-) rats (lacking the Mrp2 transporter), the clearance and hepatic extraction ratio of MPA were significantly lower compared with control Wistar rats, suggesting a difference in the capacity of the TR(-) rats to metabolize MPA in situ. There is very little information regarding the phase II metabolic capabilities of TR(-) rats; therefore, the aim of this study was to investigate the in vitro glucuronidation of MPA in Wistar and TR(-) rat liver microsomal protein. A second aim was to determine whether MPAGa, cyclosporine (CsA), and/or its metabolites AM1, AM1c, and AM9 inhibit the metabolism of MPA to MPAGe in rat liver microsomes. MPAGe formation rates by Wistar and TR(-) microsomes were 0.48 and 0.65 nmol/min/mg, respectively (p = 0.33). K(m) values for control and TR(-) microsomes were 0.47 and 0.50 mM, respectively (p = 0.81). The mean (S.E.M.) ratios of MPAGe formation by Wistar rat liver microsomes incubated with 50 microM MPA plus inhibitor versus 50 microM MPA alone were MPAGa 1.2 (0.1), CsA 0.7 (0.1) (p < 0.05), AM1 2.2 (0.3) (p < 0.05), AM1c 1.2 (0.2), and AM9 1.0 (0.2). Our results suggest that lower in situ glucuronidation of MPA in TR(-) rats may be because of inhibition of glucuronidation by endogenous and exogenous compounds that accumulate in the transporter-deficient rat. Whereas CsA inhibits glucuronidation of MPA, its metabolite AM1 enhances MPAGe formation by rat liver microsomes. PMID- 17908923 TI - Pharmacokinetics and metabolism of 14C-brivaracetam, a novel SV2A ligand, in healthy subjects. AB - This study was designed to investigate the human absorption, disposition, and mass balance of (14)C-brivaracetam, a novel high affinity SV2A ligand with potent anticonvulsant activity. Six healthy male subjects received a single p.o. dose of (14)C-brivaracetam (150 mg, 82 microCi, or 3.03 MBq). Serial blood and complete urine and feces were collected until 144 h postdose. Expired air samples were obtained until 24 h. Brivaracetam was rapidly absorbed, with C(max) of 4 mug/ml occurring within 1.5 h of dosing. Unchanged brivaracetam amounted to 90% of the total plasma radioactivity, suggesting a modest first-pass effect. Plasma protein binding of radioactivity was low (17.5%). Urinary excretion exceeded 90% after 2 days, and the final mass balance reached 96.8% of the radioactivity in urine and 0.7% in feces. Only 8.6% of the radioactive dose was recovered in urine as unchanged brivaracetam, the remainder being identified as non-cytochrome P450 (P450)- and P450-dependent biotransformation products resulting from hydrolysis of the amide moiety (M9, 34.2%), hydroxylation of the n-propyl side chain (M1b, 15.9%), and a combination of these two pathways leading to the hydroxy acid (M4b, 15.2%). Minor amounts of taurine and glucuronic acid conjugates and other oxidized derivatives were also identified. Brivaracetam is completely absorbed, is weakly bound to plasma proteins, extensively biotransformed through several metabolic pathways, and eliminated renally. PMID- 17908924 TI - Disposition and metabolism of [14C]brasofensine in rats, monkeys, and humans. AB - Brasofensine is an inhibitor of the synaptic dopamine transporter. These studies were conducted to characterize the pharmacokinetics, absolute bioavailability, disposition, and metabolism of brasofensine after i.v. and/or p.o. administrations of [(14)C]brasofensine in rats (1.5 mg/kg i.v., 4 mg/kg p.o.) and monkeys (4 mg i.v., 12 mg p.o.) and humans (50 mg p.o.). Brasofensine was rapidly absorbed after p.o. administration in rats and monkeys, with peak plasma concentrations occurring 0.5 to 1 h but 3 to 8 h for brasofensine in humans. Plasma terminal elimination half-lives were approximately 2 h in rats, approximately 4 h in monkeys, and approximately 24 h in humans. Total body clearance and steady-state volume of distribution values were 199 ml/min/kg and 24 l/kg, respectively, in the rat and 32 ml/min/kg and 46 l/kg, respectively, in the monkey. Absolute bioavailability was 7% in rats and 0.8% in monkeys. After a single p.o. dose, urinary excretion of radioactivity accounted for 20% of the administered dose in rats, 70% in monkeys, and 86% in humans, with the remainder excreted into the feces. Brasofensine had extensive first-pass metabolism following p.o. administration in humans, monkeys, and rats. It primarily underwent O- and N-demethylation and isomerization. Some of the desmethyl metabolites were further converted to glucuronides. These primary metabolites and glucuronides of demethyl brasofensine (M1 and M2) were major circulating metabolites in humans and were also observed in rat and monkey plasma. PMID- 17908925 TI - Move on up, it's time for change--mobile signals controlling photoperiod dependent flowering. AB - Plants do not bloom randomly--but how do they know when and where to make flowers? Here, we review molecular mechanisms that integrate spatial and temporal information in day-length-dependent flowering. Primarily through genetic analyses in two species, Arabidopsis thaliana and rice, we today understand the essentials of two central issues in plant biology: how the appropriate photoperiod generates an inductive stimulus based on an external coincidence mechanism, and the nature of the mobile flowering signal, florigen, which relays photoperiod-dependent information from the leaf to the growing tip of the plant, the shoot apex. PMID- 17908926 TI - Bimodal degradation of MLL by SCFSkp2 and APCCdc20 assures cell cycle execution: a critical regulatory circuit lost in leukemogenic MLL fusions. AB - Human chromosome 11q23 translocations disrupting MLL result in poor prognostic leukemias. It fuses the common MLL N-terminal approximately 1400 amino acids in frame with >60 different partners without shared characteristics. In addition to the well-characterized activity of MLL in maintaining Hox gene expression, our recent studies established an MLL-E2F axis in orchestrating core cell cycle gene expression including Cyclins. Here, we demonstrate a biphasic expression of MLL conferred by defined windows of degradation mediated by specialized cell cycle E3 ligases. Specifically, SCF(Skp2) and APC(Cdc20) mark MLL for degradation at S phase and late M phase, respectively. Abolished peak expression of MLL incurs corresponding defects in G1/S transition and M-phase progression. Conversely, overexpression of MLL blocks S-phase progression. Remarkably, MLL degradation initiates at its N-terminal approximately 1400 amino acids, and tested prevalent MLL fusions are resistant to degradation. Thus, impaired degradation of MLL fusions likely constitutes the universal mechanism underlying all MLL leukemias. Our data conclude an essential post-translational regulation of MLL by the cell cycle ubiquitin/proteasome system (UPS) assures the temporal necessity of MLL in coordinating cell cycle progression. PMID- 17908927 TI - A role for AGL ubiquitination in the glycogen storage disorders of Lafora and Cori's disease. AB - Cori's disease is a glycogen storage disorder characterized by a deficiency in the glycogen debranching enzyme, amylo-1,6-glucosidase,4-alpha-glucanotransferase (AGL). Here, we demonstrate that the G1448R genetic variant of AGL is unable to bind to glycogen and displays decreased stability that is rescued by proteasomal inhibition. AGL G1448R is more highly ubiquitinated than its wild-type counterpart and forms aggresomes upon proteasome impairment. Furthermore, the E3 ubiquitin ligase Malin interacts with and promotes the ubiquitination of AGL. Malin is known to be mutated in Lafora disease, an autosomal recessive disorder clinically characterized by the accumulation of polyglucosan bodies resembling poorly branched glycogen. Transfection studies in HepG2 cells demonstrate that AGL is cytoplasmic whereas Malin is predominately nuclear. However, after depletion of glycogen stores for 4 h, approximately 90% of transfected cells exhibit partial nuclear staining for AGL. Furthermore, stimulation of cells with agents that elevate cAMP increases Malin levels and Malin/AGL complex formation. Refeeding mice for 2 h after an overnight fast causes a reduction in hepatic AGL levels by 48%. Taken together, these results indicate that binding to glycogen crucially regulates the stability of AGL and, further, that its ubiquitination may play an important role in the pathophysiology of both Lafora and Cori's disease. PMID- 17908928 TI - Accelerated aging and failure to segregate damaged proteins in Sir2 mutants can be suppressed by overproducing the protein aggregation-remodeling factor Hsp104p. AB - The levels of oxidatively damaged, carbonylated, proteins increase with the replicative age of yeast mother cells. We show here that such carbonylated proteins are associated with Hsp104p-containing protein aggregates and that these aggregates, like oxidized proteins, are retained in the progenitor cell during cytokinesis by a Sir2p-dependent process. Deletion of HSP104 resulted in a breakdown of damage asymmetry, and overproduction of Hsp104p partially restored damage retention in sir2Delta cells, suggesting that functional chaperones associated with protein aggregates are required for the establishment of damage asymmetry and that these functions are limited in sir2Delta cells. In line with this, Hsp104p and several Hsp70s displayed elevated damaged in sir2Delta cells, and protein aggregates were rescued at a slower rate in this mutant. Moreover, overproduction of Hsp104p suppressed the accelerated aging of cells lacking Sir2p, and drugs inhibiting damage segregation further demonstrated that spatial quality control is required to rejuvenate the progeny. PMID- 17908929 TI - Lineage tracing demonstrates the venous origin of the mammalian lymphatic vasculature. AB - The origin of the mammalian lymphatic vasculature has been debated for more than 100 years. Whether lymphatic endothelial cells have a single or dual, venous or mesenchymal origin remains controversial. To resolve this debate, we performed Cre/loxP-based lineage-tracing studies using mouse strains expressing Cre recombinase under the control of the Tie2, Runx1, or Prox1 promoter elements. These studies, together with the analysis of Runx1-mutant embryos lacking definitive hematopoiesis, conclusively determined that from venous-derived lymph sacs, lymphatic endothelial cells sprouted, proliferated, and migrated to give rise to the entire lymphatic vasculature, and that hematopoietic cells did not contribute to the developing lymph sacs. We conclude that the mammalian lymphatic system has a solely venous origin. PMID- 17908930 TI - Activation of the UNC5B receptor by Netrin-1 inhibits sprouting angiogenesis. AB - Netrins are secreted molecules with roles in axonal growth and angiogenesis. The Netrin receptor UNC5B is required during embryonic development for vascular patterning, suggesting that it may also contribute to postnatal and pathological angiogenesis. Here we show that unc5b is down-regulated in quiescent adult vasculature, but re-expressed during sprouting angiogenesis in matrigel and tumor implants. Stimulation of UNC5B-expressing neovessels with an agonist (Netrin-1) inhibits sprouting angiogenesis. Genetic loss of function of unc5b reduces Netrin 1-mediated angiogenesis inhibition. Expression of UNC5B full-length receptor also triggers endothelial cell repulsion in response to Netrin-1 in vitro, whereas a truncated UNC5B lacking the intracellular signaling domain fails to induce repulsion. These data show that UNC5B activation inhibits sprouting angiogenesis, thus identifying UNC5B as a potential anti-angiogenic target. PMID- 17908931 TI - Temporal ChIP-on-chip reveals Biniou as a universal regulator of the visceral muscle transcriptional network. AB - Smooth muscle plays a prominent role in many fundamental processes and diseases, yet our understanding of the transcriptional network regulating its development is very limited. The FoxF transcription factors are essential for visceral smooth muscle development in diverse species, although their direct regulatory role remains elusive. We present a transcriptional map of Biniou (a FoxF transcription factor) and Bagpipe (an Nkx factor) activity, as a first step to deciphering the developmental program regulating Drosophila visceral muscle development. A time course of chromatin immunoprecipitatation followed by microarray analysis (ChIP on-chip) experiments and expression profiling of mutant embryos reveal a dynamic map of in vivo bound enhancers and direct target genes. While Biniou is broadly expressed, it regulates enhancers driving temporally and spatially restricted expression. In vivo reporter assays indicate that the timing of Biniou binding is a key trigger for the time span of enhancer activity. Although bagpipe and biniou mutants phenocopy each other, their regulatory potential is quite different. This network architecture was not apparent from genetic studies, and highlights Biniou as a universal regulator in all visceral muscle, regardless of its developmental origin or subsequent function. The regulatory connection of a number of Biniou target genes is conserved in mice, suggesting an ancient wiring of this developmental program. PMID- 17908932 TI - Cro's role in the CI Cro bistable switch is critical for {lambda}'s transition from lysogeny to lytic development. AB - CI represses cro; Cro represses cI. This double negative feedback loop is the core of the classical CI-Cro epigenetic switch of bacteriophage lambda. Despite the classical status of this switch, the role in lambda development of Cro repression of the P(RM) promoter for CI has remained unclear. To address this, we created binding site mutations that strongly impaired Cro repression of P(RM) with only minimal effects on CI regulation of P(RM). These mutations had little impact on lambda development after infection but strongly inhibited the transition from lysogeny to the lytic pathway. We demonstrate that following inactivation of CI by ultraviolet treatment of lysogens, repression of P(RM) by Cro is needed to prevent synthesis of new CI that would otherwise significantly impede lytic development. Thus a bistable CI-Cro circuit reinforces the commitment to a developmental transition. PMID- 17908933 TI - Defining the roles of the periplasmic chaperones SurA, Skp, and DegP in Escherichia coli. AB - Integral beta-barrel proteins (OMPs) are a major class of outer membrane proteins in Gram-negative bacteria. In Escherichia coli, these proteins are synthesized in the cytoplasm, translocated across the inner membrane via the Sec machinery, and assembled in the outer membrane through an unknown mechanism that requires the outer membrane YaeT complex and the periplasmic chaperones SurA, DegP, and Skp. Here, we have established the relationship between these three chaperones providing insight into the mechanism of OMP biogenesis using depletion analysis. Depletion of SurA alone results in a marked decrease in outer membrane density, while the loss of DegP and Skp has no effect on outer membrane composition. Furthermore, we demonstrate that SurA and YaeT interact directly in vivo. Based on these results, we suggest that SurA is the primary chaperone responsible for the periplasmic transit of the bulk mass of OMPs to the YaeT complex. The role of Skp and DegP is amplified in the absence of SurA. Evidence presented suggests that DegP/Skp function to rescue OMPs that fall off the SurA pathway. The seemingly redundant periplasmic chaperones do function in parallel, but the relative importance of the primary function of each pathway depends on whether or not cells are under stress. PMID- 17908934 TI - Telomerase repeat addition processivity is increased at critically short telomeres in a Tel1-dependent manner in Saccharomyces cerevisiae. AB - Telomerase is the ribonucleoprotein enzyme that elongates telomeres to counteract telomere shortening. The core enzyme consists of a reverse transcriptase protein subunit and an RNA subunit. The RNA subunit contains a short region that is used as a template by the reverse transcriptase to add short, tandem, G-rich repeats to the 3' ends of telomeres. By coexpressing two RNA subunits that differ in the telomeric repeat sequence specified and examining the telomere extensions after one cell cycle, we determined that Saccharomyces cerevisiae telomerase can dissociate and reassociate from a given telomere during one cell cycle. We also confirmed that telomerase is nonprocessive in terms of telomeric repeat addition. However, repeat addition processivity is significantly increased at extremely short telomeres, a process that is dependent on the ATM-ortholog Tel1. We propose that this enhancement of telomerase processivity at short telomeres serves to rapidly elongate critically short telomeres. PMID- 17908935 TI - The nature of telomere fusion and a definition of the critical telomere length in human cells. AB - The loss of telomere function can result in telomeric fusion events that lead to the types of genomic rearrangements, such as nonreciprocal translocations, that typify early-stage carcinogenesis. By using single-molecule approaches to characterize fusion events, we provide a functional definition of fusogenic telomeres in human cells. We show that approximately half of the fusion events contained no canonical telomere repeats at the fusion point; of those that did, the longest was 12.8 repeats. Furthermore, in addition to end-replication losses, human telomeres are subjected to large-scale deletion events that occur in the presence or absence of telomerase. Here we show that these telomeres are fusogenic, and thus despite the majority of telomeres being maintained at a stable length in normal human cells, a subset of stochastically shortened telomeres can potentially cause chromosomal instability. Telomere fusion was accompanied by the deletion of one or both telomeres extending several kilobases into the telomere-adjacent DNA, and microhomology was observed at the fusion points. This contrasted with telomere fusion that was observed following the experimental disruption of TRF2. The distinct error-prone mutational profile of fusion between critically shortened telomeres in human cells was reminiscent of Ku-independent microhomology-mediated end-joining. PMID- 17908936 TI - Opposing activities of two novel members of the IL-1 ligand family regulate skin inflammation. AB - The interleukin (IL)-1 family members IL-1alpha, -1beta, and -18 are potent inflammatory cytokines whose activities are dependent on heterodimeric receptors of the IL-1R superfamily, and which are regulated by soluble antagonists. Recently, several new IL-1 family members have been identified. To determine the role of one of these family members in the skin, transgenic mice expressing IL1F6 in basal keratinocytes were generated. IL1F6 transgenic mice exhibit skin abnormalities that are dependent on IL-1Rrp2 and IL-1RAcP, which are two members of the IL-1R family. The skin phenotype is characterized by acanthosis, hyperkeratosis, the presence of a mixed inflammatory cell infiltrate, and increased cytokine and chemokine expression. Strikingly, the combination of the IL-1F6 transgene with an IL1F5 deficiency results in exacerbation of the skin phenotype, demonstrating that IL-1F5 has antagonistic activity in vivo. Skin from IL1F6 transgenic, IL1F5(-/-) pups contains intracorneal and intraepithelial pustules, nucleated corneocytes, and dilated superficial dermal blood vessels. Additionally, expression of IL1RL2, -1F5, and -1F6 is increased in human psoriatic skin. In summary, dysregulated expression of novel agonistic and antagonistic IL-1 family member ligands can promote cutaneous inflammation, revealing potential novel targets for the treatment of inflammatory skin disorders. PMID- 17908937 TI - Thymic emigration revisited. AB - Conventional alphabeta T cell precursors undergo positive selection in the thymic cortex. When this is successful, they migrate to the medulla and are exposed to tissue-specific antigens (TSA) for purposes of central tolerance, and they undergo maturation to become functionally responsive T cells. It is commonly understood that thymocytes spend up to 2 wk in the medulla undergoing these final maturation steps before emigrating to peripheral lymphoid tissues. In addition, emigration is thought to occur via a stochastic mechanism whereby some progenitors leave early and others leave late-a so-called "lucky dip" process. However, recent research has revealed that medullary thymocytes are a heterogeneous mix of naive alphabeta T cell precursors, memory T cells, natural killer T cells, and regulatory T cells. Given this, we revisited the question of how long it takes naive alphabeta T cell precursors to emigrate. We combined the following three approaches to study this question: BrdU labeling, intrathymic injection of a cellular tag, and RAG2p-GFP reporter mice. We established that, on average, naive alphabeta T cell precursors emigrate only 4-5 d after becoming single-positive (SP) thymocytes. Furthermore, emigration occurs via a strict "conveyor belt" mechanism, where the oldest thymocytes leave first. PMID- 17908938 TI - Proliferative arrest and rapid turnover of thymic epithelial cells expressing Aire. AB - Expression of autoimmune regulator (Aire) by thymic medullary epithelial cells (MECs) is critical for central tolerance of self. To explore the mechanism by which such a rare cell population imposes tolerance on the large repertoire of differentiating thymocytes, we examined the proliferation and turnover of Aire(+) and Aire(-) MEC subsets through flow cytometric analysis of 5-bromo 2'deoxyuridine (BrdU) incorporation. The Aire(+) MEC subset was almost entirely postmitotic and derived from cycling Aire(-) precursors. Experiments using reaggregate thymic organ cultures revealed the presence of such precursors among Aire(-) MECs expressing low levels of major histocompatibility complex class II and CD80. The kinetics of BrdU decay showed the Aire(+) population to have a high turnover. Aire did not have a direct impact on the division of MECs in vitro or in vivo but, rather, induced their apoptosis. We argue that these properties strongly favor a "terminal differentiation" model for Aire function in MECs, placing strict temporal limits on the operation of any individual Aire(+) MEC in central tolerance induction. We further speculate that the speedy apoptosis of Aire-expressing MECs may be a mechanism to promote cross-presentation of the array of peripheral-tissue antigens they produce. PMID- 17908939 TI - Ralph Steinman: dendritic cells bring home the Lasker. AB - Ralph Steinman is perhaps best known as a codiscoverer of dendritic cells (DCs) and as a founding father of the research area that these cells have spawned. For his discovery, Steinman was recently awarded the 2007 Albert Lasker Award for Basic Medical Research. Yet the man behind the research holds his praise for the many other scientists-in the U.S. and abroad-who have further advanced the therapeutic promise of DCs. PMID- 17908940 TI - National and regional assessment of antimicrobial resistance among community acquired respiratory tract pathogens identified in a 2005-2006 U.S. Faropenem surveillance study. AB - Surveillance studies conducted in the United States over the last decade have revealed increasing resistance among community-acquired respiratory pathogens, especially Streptococcus pneumoniae, that may limit future options for empirical therapy. The objective of this study was to assess the scope and magnitude of the problem at the national and regional levels during the 2005-2006 respiratory season (the season when community-acquired respiratory pathogens are prevalent) in the United States. Also, since faropenem is an oral penem being developed for the treatment of community-acquired respiratory tract infections, another study objective was to provide baseline data to benchmark changes in the susceptibility of U.S. respiratory pathogens to the drug in the future. The in vitro activities of faropenem and other agents were determined against 1,543 S. pneumoniae isolates, 978 Haemophilus influenzae isolates, and 489 Moraxella catarrhalis isolates collected from 104 U.S. laboratories across six geographic regions during the 2005-2006 respiratory season. Among S. pneumoniae isolates, the rates of resistance to penicillin, amoxicillin-clavulanate, and cefdinir were 16, 6.4, and 19.2%, respectively. The least effective agents were trimethoprim sulfamethoxazole (SXT) and azithromycin, with resistance rates of 23.5 and 34%, respectively. Penicillin resistance rates for S. pneumoniae varied by region (from 8.7 to 22.5%), as did multidrug resistance rates for S. pneumoniae (from 8.8 to 24.9%). Resistance to beta-lactams, azithromycin, and SXT was higher among S. pneumoniae isolates from children than those from adults. beta-Lactamase production rates among H. influenzae and M. catarrhalis isolates were 27.4 and 91.6%, respectively. Faropenem MICs at which 90% of isolates are inhibited were 0.5 mug/ml for S. pneumoniae, 1 mug/ml for H. influenzae, and 0.5 mug/ml for M. catarrhalis, suggesting that faropenem shows promise as a treatment option for respiratory infections caused by contemporary resistant phenotypes. PMID- 17908941 TI - Rifampin inhibits prostaglandin E2 production and arachidonic acid release in human alveolar epithelial cells. AB - Rifampin, a potent antimicrobial agent, is a major drug in the treatment of tuberculosis. There is evidence that rifampin also serves as an immunomodulator. Based on findings that arachidonic acid and its metabolites are involved in the pathogeneses of Mycobacterium tuberculosis infections, we investigated whether rifampin affects prostaglandin E(2) (PGE(2)) production in human alveolar epithelial cells stimulated with interleukin-1beta. Rifampin caused a dose dependent inhibition of PGE(2) production. At doses of 100, 50, and 25 microg/ml, it inhibited PGE(2) production by 75%, 59%, and 45%, respectively (P < 0.001). Regarding the mechanism involved, rifampin caused a time- and dose-dependent inhibition of arachidonic acid release from the alveolar cells. At doses of 100, 50, 25, and 10 mug/ml, it significantly inhibited the release of arachidonic acid by 93%, 64%, 58%, and 35%, respectively (P < 0.001). Rifampin did not affect the phosphorylation of cytosolic phospholipase A(2) or the expression of cyclooxygenase-2. The inhibition of PGE(2), and presumably other arachidonic acid products, probably contributes to the efficacy of rifampin in the treatment of tuberculosis and may explain some of its adverse effects. PMID- 17908943 TI - A MurF inhibitor that disrupts cell wall biosynthesis in Escherichia coli. AB - MurF is an essential enzyme of bacterial cell wall biosynthesis. Few MurF inhibitors have been reported, and none have displayed measurable antibacterial activity. Through the use of a MurF binding assay, a series of 8 hydroxyquinolines that bound to the Escherichia coli enzyme and inhibited its activity was identified. To derive additional chemotypes lacking 8 hydroxyquinoline, a known chelating moiety, a pharmacophore model was constructed from the series and used to select compounds for testing in the MurF binding and enzymatic inhibition assays. Whereas the original diverse library yielded 0.01% positive compounds in the binding assay, of which 6% inhibited MurF enzymatic activity, the pharmacophore-selected set yielded 14% positive compounds, of which 37% inhibited the enzyme, suggesting that the model enriched for compounds with affinity to MurF. A 4-phenylpiperidine (4-PP) derivative identified by this process displayed antibacterial activity (MIC of 8 microg/ml against permeable E. coli) including cell lysis and a 5-log(10)-unit decrease in CFU. Importantly, treatment of E. coli with 4-PP resulted in a 15-fold increase in the amount of the MurF UDP-MurNAc-tripeptide substrate, and a 50% reduction in the amount of the MurF UDP-MurNAc-pentapeptide product, consistent with inhibition of the MurF enzyme within bacterial cells. Thus, 4-PP is the first reported inhibitor of the MurF enzyme that may contribute to antibacterial activity by interfering with cell wall biosynthesis. PMID- 17908944 TI - Antileishmanial structure-activity relationships of synthetic phospholipids: in vitro and in vivo activities of selected derivatives. AB - Antileishmanial activities of 91 synthetic phospholipids against Leishmania donovani were evaluated in vitro and cytotoxicity assessed against two mammalian cell lines. Promising compounds were tested further in vivo. In vitro structure activity relationships showed a positive contribution of head groups bearing ring systems (N-methylpiperidino and N-methylmorpholino) to antileishmanial activity. PMID- 17908942 TI - Frequency of development and associated physiological cost of azithromycin resistance in Chlamydia psittaci 6BC and C. trachomatis L2. AB - Azithromycin is a major drug used in the treatment and prophylaxis of chlamydial infections. Spontaneous azithromycin-resistant mutants of Chlamydia psittaci 6BC were isolated in vitro in the plaque assay at a frequency of about 10(-8). Isogenic clonal variants with A(2058)C, A(2059)G, or A(2059)C mutations in the unique 23S rRNA gene (Escherichia coli numbering system) displayed MICs for multiple macrolides (i.e., azithromycin, erythromycin, josamycin, and spiramycin) at least 100 times higher than those of the parent strain and were also more resistant to the lincosamide clindamycin. Chlamydia trachomatis L2 variants with a Gln-to-Lys substitution in ribosomal protein L4 at position 66 (E. coli numbering system), conferring an eightfold decrease in azithromycin and erythromycin sensitivities and a fourfold decrease in josamycin and spiramycin sensitivities, were isolated following serial passage in subinhibitory concentrations of azithromycin. Each mutation was stably maintained in the absence of selection but severely affected chlamydial infectivity, as determined by monitoring the development of each isolate over 46 h in the absence of selection, in pure culture or in 1:1 competition with the isogenic parent. Data in this study support the hypothesis that the mechanisms which confer high-level macrolide resistance in chlamydiae carry a prohibitive physiological cost and may thus limit the emergence of highly resistant clones of these important pathogens in vivo. PMID- 17908945 TI - Outbreaks associated with contaminated antiseptics and disinfectants. PMID- 17908947 TI - In vitro evaluation of the activities of telavancin, cefazolin, and vancomycin against methicillin-susceptible and methicillin-resistant Staphylococcus aureus in peritoneal dialysate. AB - This study compared the ability of telavancin to the ability of cefazolin and vancomycin to eliminate staphylococci from peritoneal dialysis fluid by using a static in vitro model to simulate the conditions of peritoneal dialysis. The results showed that telavancin exhibited statistically significantly better kill (P < 0.05) against both methicillin-susceptible and methicillin-resistant Staphylococcus aureus. PMID- 17908946 TI - Antimicrobial drug resistance of Salmonella enterica serovar typhi in asia and molecular mechanism of reduced susceptibility to the fluoroquinolones. AB - This study describes the pattern and extent of drug resistance in 1,774 strains of Salmonella enterica serovar Typhi isolated across Asia between 1993 and 2005 and characterizes the molecular mechanisms underlying the reduced susceptibilities to fluoroquinolones of these strains. For 1,393 serovar Typhi strains collected in southern Vietnam, the proportion of multidrug resistance has remained high since 1993 (50% in 2004) and there was a dramatic increase in nalidixic acid resistance between 1993 (4%) and 2005 (97%). In a cross-sectional sample of 381 serovar Typhi strains from 8 Asian countries, Bangladesh, China, India, Indonesia, Laos, Nepal, Pakistan, and central Vietnam, collected in 2002 to 2004, various rates of multidrug resistance (16 to 37%) and nalidixic acid resistance (5 to 51%) were found. The eight Asian countries involved in this study are home to approximately 80% of the world's typhoid fever cases. These results document the scale of drug resistance across Asia. The Ser83-->Phe substitution in GyrA was the predominant alteration in serovar Typhi strains from Vietnam (117/127 isolates; 92.1%). No mutations in gyrB, parC, or parE were detected in 55 of these strains. In vitro time-kill experiments showed a reduction in the efficacy of ofloxacin against strains harboring a single-amino acid substitution at codon 83 or 87 of GyrA; this effect was more marked against a strain with a double substitution. The 8-methoxy fluoroquinolone gatifloxacin showed rapid killing of serovar Typhi harboring both the single- and double-amino acid substitutions. PMID- 17908948 TI - In Vitro susceptibility of methicillin-resistant Staphylococcus aureus and methicillin-susceptible Staphylococcus aureus to a new antimicrobial, copper silicate. AB - The soluble copper silicate (CS) MIC of 100 strains of methicillin-resistant Staphylococcus aureus and 100 strains of methicillin-susceptible S. aureus (MSSA) was 175 mg Cu/liter. Bactericidal and postantibiotic effects (> or =1 h) were seen at 2x MIC and 4x MIC. The frequency of mutation was <10(-9), and serial passage could not extend growth beyond 1.6x MIC. PMID- 17908949 TI - Effect of silver content on the structure and antibacterial activity of silver doped phosphate-based glasses. AB - Staphylococcus aureus can cause a range of diseases, such as osteomyelitis, as well as colonize implanted medical devices. In most instances the organism forms biofilms that not only are resistant to the body's defense mechanisms but also display decreased susceptibilities to antibiotics. In the present study, we have examined the effect of increasing silver contents in phosphate-based glasses to prevent the formation of S. aureus biofilms. Silver was found to be an effective bactericidal agent against S. aureus biofilms, and the rate of silver ion release (0.42 to 1.22 microg x mm(-2) x h(-1)) from phosphate-based glass was found to account for the variation in its bactericidal effect. Analysis of biofilms by confocal microscopy indicated that they consisted of an upper layer of viable bacteria together with a layer ( approximately 20 microm) of nonviable cells on the glass surface. Our results showed that regardless of the silver contents in these glasses (10, 15, or 20 mol%) the silver exists in its +1 oxidation state, which is known to be a highly effective bactericidal agent compared to that of silver in other oxidation states (+2 or +3). Analysis of the glasses by (31)P nuclear magnetic resonance imaging and high-energy X-ray diffraction showed that it is the structural rearrangement of the phosphate network that is responsible for the variation in silver ion release and the associated bactericidal effectiveness. Thus, an understanding of the glass structure is important in interpreting the in vitro data and also has important clinical implications for the potential use of the phosphate-based glasses in orthopedic applications to deliver silver ions to combat S. aureus biofilm infections. PMID- 17908950 TI - Activity of a potent hepatitis C virus polymerase inhibitor in the chimpanzee model. AB - A-837093 is a potent and specific nonnucleoside inhibitor of the hepatitis C virus (HCV) nonstructural protein 5B (NS5B) RNA-dependent RNA polymerase. It possesses nanomolar potencies in both enzymatic and replicon-based cell culture assays. In rats and dogs this compound demonstrated an oral plasma half-life of greater than 7 h, and its bioavailability was >60%. In monkeys it had a half-life of 1.9 h and 15% bioavailability. Its antiviral efficacy was evaluated in two chimpanzees infected with HCV in a proof-of-concept study. The design included oral dosing of 30 mg per kg of body weight twice a day for 14 days, followed by a 14-day posttreatment observation. Maximum viral load reductions of 1.4 and 2.5 log(10) copies RNA/ml for genotype 1a- and 1b-infected chimpanzees, respectively, were observed within 2 days after the initiation of treatment. After this initial drop in the viral load, a rebound of plasma HCV RNA was observed in the genotype 1b-infected chimpanzee, while the genotype 1a-infected chimpanzee experienced a partial rebound that lasted throughout the treatment period. Clonal analysis of NS5B gene sequences derived from the plasma of A-837093-treated chimpanzees revealed the presence of several mutations associated with resistance to A 837093, including Y448H, G554D, and D559G in the genotype 1a-infected chimpanzee and C316Y and G554D in the genotype 1b-infected chimpanzee. The identification of resistance-associated mutations in both chimpanzees is consistent with the findings of in vitro selection studies, in which many of the same mutations were selected. These findings validate the antiviral efficacy and resistance development of benzothiadiazine HCV polymerase inhibitors in vivo. PMID- 17908951 TI - In vitro resistance study of rupintrivir, a novel inhibitor of human rhinovirus 3C protease. AB - Rupintrivir (formerly AG7088) is an irreversible inhibitor of the human rhinovirus (HRV) 3C protease that has been demonstrated to have in vitro activity against all HRVs tested, consistent with its interaction with a strictly conserved subset of amino acids in the 3C protease. The potential for resistance was studied following in vitro serial passage of HRV serotypes 14, 2, 39, and Hanks in the presence of increasing rupintrivir concentrations. HRV variants with reduced susceptibilities to rupintrivir (sevenfold for HRV 14) or with no significant reductions in susceptibility but genotypic changes (HRV 2, 39, and Hanks) were initially isolated following 14 to 40 cumulative days in culture (three to six passages). Sequence analysis of the 3C protease identified one to three substitutions in diverse patterns but with common features (T129T/A, T131T/A, and T143P/S in HRV 14; N165T in HRV 2; N130N/K and L136L/F in HRV 39; T130A in HRV Hanks). Notably, three of the four HRV variants contained a substitution at residue 130 (residue 129 in HRV 14). Continued selection in the presence of escalating concentrations of rupintrivir (40 to 72 days) resulted in the accumulation of additional mutations (A121A/V and Y139Y/H in HRV 14, E3E/G and A103A/V in HRV 2, S105T in HRV 39), with only minimal further reductions in susceptibility (up to fivefold). The ability of specific substitutions to confer resistance was examined by susceptibility testing of HRV 14 variants constructed to contain 3C protease mutations. In summary, the slow accumulation of multiple amino acid substitutions with only minimal to moderate reductions in susceptibility highlight the advantages of 3C protease as an antiviral target. PMID- 17908952 TI - "PIKing" the winner for phosphatidylinositol 3-kinase inhibitors in ErbB2 positive breast cancer: let's not "PTENed" it's easy! PMID- 17908954 TI - Targeting the cytoplasmic and nuclear functions of signal transducers and activators of transcription 3 for cancer therapy. AB - Signal transducers and activators of transcription (STAT) are a highly conserved family of transcription factors that are activated by phosphorylation in the cytoplasm, after which they translocate to the nucleus to regulate gene expression. Among the seven STATs, STAT3 is of particular interest due to its constitutive phosphorylation in a large proportion of human cancers and its ability to induce neoplastic transformation. Inhibition of STAT3 can reverse tumor growth in experimental systems while having few effects in normal cells. These findings have implicated STAT3 as a potentially important target for therapeutic intervention. In addition to its well-described role as a transcription factor, STAT3 has been found recently to have important effects in the cytoplasm. Collectively, these functions of STAT3 directly contribute to tumorigenesis, invasion, and metastasis. Given the potential importance of STAT3 as a target for cancer therapy, molecules have been developed that can block STAT3 function at a variety of steps. These drugs show promise as anticancer agents in model systems of a variety of common human cancers. Thus, elucidating the functions of STAT3 and developing agents to inhibit this protein remain important scientific and clinical challenges. PMID- 17908953 TI - NF-kappaB gene signatures and p53 mutations in head and neck squamous cell carcinoma. PMID- 17908955 TI - Hypoxia inducible factor-1 independent pathways in tumor angiogenesis. AB - Among the factors that can stimulate angiogenesis, vascular endothelial growth factor has emerged as one of the most important, and inhibition of vascular endothelial growth factor has recently shown efficacy in the treatment of advanced colorectal cancer. Hypoxia develops within solid tumors and is one of the most potent stimuli of vascular endothelial growth factor expression. This effect is mediated primarily by hypoxia inducible factor-1 (HIF-1), often considered a master regulator of angiogenesis in hypoxia. Consequently, inhibition of HIF-1 has been proposed as a strategy to block tumor angiogenesis therapeutically. However, accumulating evidence indicates that HIF-independent pathways can also control angiogenesis. This review highlights some of the key signaling pathways independent of HIF-1 that can stimulate angiogenesis in hypoxia. Understanding the full spectrum of molecular pathways that control tumor angiogenesis is critical for the optimal design of targeted therapies. PMID- 17908956 TI - Universal and stemness-related tumor antigens: potential use in cancer immunotherapy. PMID- 17908957 TI - A novel nuclear factor-kappaB gene signature is differentially expressed in head and neck squamous cell carcinomas in association with TP53 status. AB - PURPOSE: To determine if gene signatures differentially expressed in head and neck squamous cell carcinomas (HNSCC) are related to alterations in transcription factors nuclear factor-kappaB (NF-kappaB) and TP53 previously associated with decreased cell death, response to therapy, and worse prognosis. EXPERIMENTAL DESIGN: Unique gene signatures expressed by HNSCC lines were identified by cDNA microarray, principal components, and cluster analyses and validated by quantitative reverse transcription-PCR (RT-PCR) and in situ hybridization. Bioinformatic analysis of the promoters and ontogeny of these clustered genes was done. Expression of proteins encoded by genes of a putative NF-kappaB signature, NF-kappaB p65, and TP53 were examined in HNSCC tissue specimens by immunostaining. Predicted promoter binding and modulation of expression of candidate NF-kappaB genes and cell survival were evaluated by p65 chromatin immunoprecipitation (ChIP) and small interfering RNA (siRNA) knockdown. RESULTS: Two groups of HNSCC exhibiting distinct gene signatures were identified: cluster A enriched for histone genes, with a higher prevalence of TP53 promoter binding motifs; and cluster B enriched for injury response genes with NF-kappaB regulatory motifs. Coexpression of cluster B proteins was observed with strong NF kappaB phospho-p65 and weak TP53 staining, and NF-kappaB phospho-p65 was inversely associated with TP53 (P = 0.02). Promoter binding of the NF-kappaB signature genes was confirmed by p65 ChIP, and down-modulation of their expression and cell death were induced by p65 siRNA. CONCLUSION: NF-kappaB promotes expression of a novel NF-kappaB-related gene signature and cell survival in HNSCC that weakly express TP53, a subset previously associated with inactivated wild-type TP53, greater resistance to chemoradiotherapy, and worse prognosis. PMID- 17908958 TI - Smad3 is overexpressed in advanced human prostate cancer and necessary for progressive growth of prostate cancer cells in nude mice. AB - PURPOSE: The purpose of this study was to investigate the potential role of Smad3, a key mediator of transforming growth factor-beta signaling, in progression of prostate cancer. EXPERIMENTAL DESIGN: Expression of Smad proteins was determined in human prostate cancer tissue array and cell lines. Growth and metastasis of cells overexpressing dominant-negative Smad3 (Smad3D) were studied to determine its role in tumor progression in mice. Cell growth, apoptosis, and expression of angiogenic molecules in tumor lesions were studied to determine potential pathways that Smad3 promotes tumor progression. RESULTS: Smad3 was overexpressed in human prostate cancer, which correlated with Gleason score and expression of proliferating cell nuclear antigen. Androgen-independent PC-3MM2 and DU145 cells expressed much higher levels of Smad3 than did androgen-dependent LNCaP, 22Rv1, and LAPC-4 cells. Overexpression of Smad3D in PC-3MM2 cells (PC 3MM2-Smad3D) had minimal direct effects on cell growth but attenuated effects of transforming growth factor-beta1 on gene expression and cell growth. Overexpression of Smad3D did not significantly alter tumor incidence but reduced tumor growth rate and metastasis incidence. Most cells in the control tumors, but not PC-3MM2-Smad3D tumors, were positively stained by an antibody to proliferating cell nuclear antigen. Microvessels and expression of angiogenic molecule interleukin-8 were significantly reduced in tumors from PC-3MM2-Smad3D cells. PC-3MM2-Smad3D tumors also expressed lower levels of vascular endothelial growth factor and platelet-derived growth factor. CONCLUSIONS: These data suggest that Smad3, through regulating angiogenic molecule expression in tumor cells, is critical for progression of human prostate cancer. PMID- 17908959 TI - Identification of Toll-like receptor 3 as a potential therapeutic target in clear cell renal cell carcinoma. AB - PURPOSE: Renal cell carcinoma (RCC) is one of the most drug-refractory cancers. The aim of this study is to discover a novel therapeutic target molecule for clear cell RCC (CCRCC), which accounts for the majority of RCC. EXPERIMENTAL DESIGN: Gene expression profiles of 27 CCRCCs and 9 normal kidney tissues as well as 15 various adult normal tissues were examined by Affymetrix U133 Plus 2.0 arrays. Among the 34 genes specifically up-regulated in CCRCC, overexpression of Toll-like receptor 3 (TLR3) mRNA and its protein was validated by quantitative reverse transcription-PCR, immunoblot, and immunohistochemistry. The effects of TLR3 signaling on in vitro cell growth were examined. RESULTS: TLR3 gene was highly expressed in CCRCC, with only limited expression in a panel of normal tissues. On immunohistochemical analysis using a monoclonal antibody against TLR3, overexpression of TLR3 was observed in 139 of 189 (73.5%) cases of CCRCC as well as in lung metastatic CCRCC (6 of 8), whereas TLR3 expression was entirely absent in chromophobe RCC (0 of 8). Polyinosinic-polycytidilic acid, a TLR3 ligand, exerted a growth-inhibitory effect against RCC cells in a TLR3-dependent manner. Moreover, a combination of polyinosinic-polycytidilic acid and IFNalpha exerted a synergistic growth-inhibitory effect against Caki-1 RCC cells. CONCLUSIONS: This is the first report that TLR3 is overexpressed in CCRCC. These observations suggest that TLR3 pathway may represent a novel therapeutic target in CCRCC. PMID- 17908960 TI - The proangiogenic phenotype of tumor-derived endothelial cells is reverted by the overexpression of platelet-activating factor acetylhydrolase. AB - PURPOSE: We previously reported that human tumor-derived endothelial cells (TEC) have an angiogenic phenotype related to the autocrine production of several angiogenic factors. The purpose of the present study was to evaluate whether an enhanced synthesis of platelet-activating factor (PAF) might contribute to the proangiogenic characteristics of TEC and whether its inactivation might inhibit angiogenesis. EXPERIMENTAL DESIGN: To address the potential role of PAF in the proangiogenic characteristics of TEC, we engineered TEC to stably overexpress human plasma PAF-acetylhydrolase (PAF-AH), the major PAF-inactivating enzyme, and we evaluated in vitro and in vivo angiogenesis. RESULTS: TECs were able to synthesize a significantly enhanced amount of PAF compared with normal human microvascular endothelial cells when stimulated with thrombin, vascular endothelial growth factor, or soluble CD154. Transfection of TEC with PAF-AH (TEC PAF-AH) significantly inhibited apoptosis resistance and spontaneous motility of TEC. In addition, PAF and vascular endothelial growth factor stimulation enhanced the motility and adhesion of TEC but not of TEC-PAF-AH. In vitro, TEC-PAF-AH lost the characteristic ability of TEC to form vessel-like structures when plated on Matrigel. Finally, when cells were injected s.c. within Matrigel in severe combined immunodeficiency mice or coimplanted with a renal carcinoma cell line, the overexpression of PAF-AH induced a significant reduction of functional vessel formation. CONCLUSIONS: These results suggest that inactivation of PAF, produced by TEC, by the overexpression of plasma PAF-AH affects survival, migration, and the angiogenic response of TEC both in vitro and in vivo. PMID- 17908961 TI - Eotaxin-2 and colorectal cancer: a potential target for immune therapy. AB - PURPOSE: To study the production of chemokines by colorectal hepatic metastases. EXPERIMENTAL DESIGN: Biopsies of resected colorectal hepatic metastases and nonneoplastic adjacent liver tissue were screened for chemokines using protein arrays and results were confirmed by ELISA and immunohistochemistry. RESULTS: Two chemokines, eotaxin-2 and MCP-1, were found at elevated levels within the tumor biopsy compared with adjacent liver. The relative increase in expression from tumor was much higher for eotaxin-2 than MCP-1, with 10 of 25 donors having a >100-fold increase in expression compared with 0 of 24 donors for MCP-1. In a parallel analysis, eotaxin-2 was also found at elevated levels in the tumor region of primary colorectal cancer biopsies. Immunohistochemical staining indicated that carcinoembryonic antigen-positive tumor cells stained strongly for eotaxin-2, implicating these cells as the predominant source of the chemokine. In vitro studies confirmed that several colorectal tumor lines produce eotaxin-2 and that secretion of this chemokine could be depressed by IFN-gamma and enhanced by the Th2-type cytokines interleukin-4 and interleukin-13. Jurkat T cells were engineered to express the receptor for eotaxin-2 (CCR3). These cells effectively migrated in response to eotaxin-2 protein, suggesting that immune cells gene modified to express a chemokine receptor may have improved abilities to home to tumor. CONCLUSIONS: Taken together, these observations confirm eotaxin-2 as a chemokine strongly associated with primary and metastatic tumors of colorectal origin. Furthermore, the importance of this result may be a useful tool in the development of targeted therapeutic approaches to colorectal tumors. PMID- 17908962 TI - Molecular analysis of colorectal cancer tumors from patients with mismatch repair proficient hereditary nonpolyposis colorectal cancer suggests novel carcinogenic pathways. AB - PURPOSE: A subset of colorectal cancers (CRC) arises in families that, despite fulfilling clinical criteria for hereditary nonpolyposis colorectal cancer (HNPCC), do not show evidence of a mismatch repair (MMR) deficiency. The main objective of this study was to characterize these tumors at the molecular level. EXPERIMENTAL DESIGN: After comprehensive germ line mutation scanning, microsatellite analysis, and MMR protein expressions, we selected a well-defined cohort of 57 colorectal tumors with no evidence of MMR defects. In this group of tumors, we analyzed KRAS, BRAF, and APC somatic mutations, as well as methylguanine methyltransferase (MGMT) and beta-catenin expression. We correlated these alterations with clinicopathologic data and explored the relationship between KRAS G > A transitions and lack of MGMT expression. RESULTS: The mutation profile at the RAS/RAF/MAPK pathway mimics sporadic microsatellite-stable CRCs. We found an average age of diagnosis 10 years older in KRAS-mutated patients (P = 0.001). In addition, we show that KRAS G > A transitions are actively selected by tumors, regardless of MGMT status. Similarities with HNPCC high-microsatellite instability tumors are observed when APC data are analyzed. The APC mutation rate was low and small insertions/deletions accounted for 70% of the alterations. In addition, we found a low frequency of beta-catenin nuclear staining. Finally, we did not find evidence of tumors arising in individuals from the same family sharing molecular features. CONCLUSIONS: We show evidence that CRC tumors arising in HNPCC families without MMR alterations have distinctive molecular features. Overall, our work shows that systematic analysis of somatic alterations in a well defined subset of CRCs is a good approach to provide new insights into the mechanisms of colorectal carcinogenesis. PMID- 17908963 TI - Comparative analysis of peritoneum and tumor eicosanoids and pathways in advanced ovarian cancer. AB - PURPOSE: To describe the eicosanoid profile and differentially expressed eicosanoid and arachidonic acid pathway genes in tissues from patients with advanced epithelial ovarian cancer (EOC). EXPERIMENTAL DESIGN: We first employed electrospray tandem mass spectrometry to determine tissue-specific concentrations of the eicosanoids prostaglandin E2 (PGE2), the hydroxyeicosatetraenoic acids (12 HETE and 5-HETE), and leukotriene (LTB4), selected for tumor growth potential, and two other bioactive lipids (15-HETE and 13-HODE) with tumor cell proliferation interference potential. The cellular location of eicosanoid activity was identified by immunofluorescence antibody costaining and confocal microscopy. Differential analysis of eicosanoid and arachidonic pathway genes was done using a previously validated cDNA microarray platform. Tissues used included EOC tumor, tumor-free malignant peritoneum (MP), and benign peritoneum (BP) from patients with benign pelvic disease. RESULTS: (a) Eicosanoid products were detected in tumor, MP, and BP specimens. PGE2 levels were significantly elevated in tumors in an overall comparison with MP or BP (P < 0.001). Combined levels of PGE2, 12-HETE, 5-HETE, and LTB4 increased progressively from low to high concentrations in BP, MP, and tumors (P = 0.012). Neither 15-HETE nor 13-HODE showed a significant opposite trend toward levels found in BP. (b) Tissue specimens representing common EOC histotypes showed strong coexpressions of cyclooxygenases (COX-1) and prostaglandin E synthases (PGES-1) on tumor cells, whereas intratumoral or peritumoral MO/MA coexpressed COX-1 and COX-2 and PGES-1 and PGES-2, respectively. (c) cDNA microarray analysis of MP, BP, and tumor showed that a number of eicosanoid and arachidonic acid pathway genes were differentially expressed in MP and BP compared with tumor, except for CYP2J2, which was increased in tumors. CONCLUSIONS: Elevated levels of eicosanoid metabolites in tumors and differential expression of eicosanoid and arachidonic acid pathway genes in the peritoneum support the involvement of bioactive lipids in the inflammatory tumor environment of EOC. PMID- 17908964 TI - Amplification of PVT1 contributes to the pathophysiology of ovarian and breast cancer. AB - PURPOSE: This study was designed to elucidate the role of amplification at 8q24 in the pathophysiology of ovarian and breast cancer because increased copy number at this locus is one of the most frequent genomic abnormalities in these cancers. EXPERIMENTAL DESIGN: To accomplish this, we assessed the association of amplification at 8q24 with outcome in ovarian cancers using fluorescence in situ hybridization to tissue microarrays and measured responses of ovarian and breast cancer cell lines to specific small interfering RNAs against the oncogene MYC and a putative noncoding RNA, PVT1, both of which map to 8q24. RESULTS: Amplification of 8q24 was associated with significantly reduced survival duration. In addition, small interfering RNA-mediated reduction in either PVT1 or MYC expression inhibited proliferation in breast and ovarian cancer cell lines in which they were both amplified and overexpressed but not in lines in which they were not amplified/overexpressed. Inhibition of PVT1 expression also induced a strong apoptotic response in cell lines in which it was overexpressed but not in lines in which it was not amplified/overexpressed. Inhibition of MYC, on the other hand, did not induce an apoptotic response in cell lines in which MYC was amplified and overexpressed. CONCLUSIONS: These results suggest that MYC and PVT1 contribute independently to ovarian and breast pathogenesis when overexpressed because of genomic abnormalities. They also suggest that PVT1-mediated inhibition of apoptosis may explain why amplification of 8q24 is associated with reduced survival duration in patients treated with agents that act through apoptotic mechanisms. PMID- 17908965 TI - CMTM5 exhibits tumor suppressor activities and is frequently silenced by methylation in carcinoma cell lines. AB - PURPOSE: CMTM5 (CKLF-like MARVEL transmembrane domain containing member 5) is located at 14q11.2, a locus associated with multiple cancers. It has six RNA splicing variants with CMTM5-v1 as the major one. We explored its expression pattern in normal tissues and tumor cell lines, as well as its functions in carcinoma cells. EXPERIMENTAL DESIGN: We evaluated CMTM5 expression by semiquantitative reverse transcription-PCR (RT-PCR) in normal tissues and carcinoma cell lines of cervical, breast, nasopharyngeal, lung, hepatocellular, esophageal, gastric, colon, and prostate. We further examined CMTM5 promoter methylation in these cell lines. We also analyzed CMTM5 expression after 5-aza-2' deoxycytidine treatment and genetic demethylation and the functional consequences of restoring CMTM5 in HeLa and PC-3 cells. RESULTS: CMTM5-v1 is broadly expressed in human normal adult and fetal tissues, but undetectable or down-regulated in most carcinoma cell lines. Its promoter methylation was detected in virtually all the silenced or down-regulated cell lines. The silencing of CMTM5 could be reversed by pharmacologic demethylation or genetic double-knockout of DNMT1 and DNMT3B, indicating methylation-mediated mechanism. Restoration of CMTM5-v1 suppressed carcinoma cell proliferation, migration, and invasion. CONCLUSIONS: These results indicate that CMTM5 exhibits tumor suppressor activities, but with frequent epigenetic inactivation in carcinoma cell lines. PMID- 17908966 TI - The impact of sex and smoking status on the mutational spectrum of epidermal growth factor receptor gene in non small cell lung cancer. AB - PURPOSE: Mutation of epidermal growth factor receptor (EGFR) gene has been reported to be present in non-small cell lung cancer (NSCLC) and significantly associated with female sex and never-smoking status. In this study, we extensively investigated the impact of sex and smoking on the EGFR mutation. EXPERIMENTAL DESIGN: We examined EGFR exons 18 to 21 status in 1,467 NSCLC patients by direct sequencing to study the impact of sex and smoking status on the EGFR mutational spectrum. RESULTS: Among 1,467 patients, 197 mutations were found at exon 19, 176 at exon 21, 21 at exon 18, and 24 at exon 20. To examine the independent effect of sex and smoking, the mutational status of each exon was compared between smokers and never smokers in each sex and between males and females stratified by smoking status. In females, exon 19 (P = 0.001) and exon 21 (P < 0.001) mutations were significantly less frequent in ever smokers compared with never smokers. In males, exon 19 (P < 0.001), exon 21 (P < 0.001), and exon 18 (P = 0.003) mutations were significantly less frequent in ever smokers compared with never smokers. In analysis stratified by smoking, there was no difference in sex among never smokers. However, exon 19 mutations were significantly less frequent in males compared with females among ever smokers (P = 0.003). In addition, the interactive effect of male sex and ever smoking status significantly decreased the frequency of exon 19 mutations (P = 0.047) when female never smoker was set as a reference. CONCLUSION: Both sex and smoking status could influence the EGFR mutational spectrum. Our findings suggest that individual EGFR exons may have differing susceptibilities for mutagenesis. PMID- 17908967 TI - Phosphorylation of estrogen receptor-alpha at Ser167 is indicative of longer disease-free and overall survival in breast cancer patients. AB - PURPOSE: Ser(167) was first identified as a major phosphorylation site of the estrogen receptor -alpha (ER) positive in the MCF7 breast cancer cell line. Subsequent studies have shown that Ser(167) phosphorylation is important in the regulation of ER activity and have identified p90RSK and AKT as protein kinases that phosphorylate Ser(167). The purpose of this study was to determine the importance of Ser(167) phosphorylation in breast cancer progression. EXPERIMENTAL DESIGN: Immunohistochemical staining of primary breast cancer biopsies (n = 290) was carried out using antibodies specific for ER phosphorylated at Ser(167) and for phosphorylated p44/p42 mitogen-activated protein kinase (MAPK), phosphorylated p90RSK, and phosphorylated AKT. RESULTS: In ER-positive breast cancer patients, Ser(167) phosphorylation was associated with low tumor grade (P = 0.011), lymph node negativity (P = 0.034), and relapse-free (P = 0.006) and overall (P = 0.023) survival. Further, Ser(167) phosphorylation was strongly associated with phosphorylated p90RSK (P < 0.001), previously shown to phosphorylate Ser(167) in vitro, as well as being associated with phosphorylated MAPK (P < 0.0005). The activities of both kinases also seemed to be indicative of better prognosis. There was, however, no association between HER2 positivity and Ser(167) phosphorylation nor were the activities of MAPK or p90RSK associated with HER2 status, suggesting that other cell surface receptors may be important in regulating these activities in breast cancer. CONCLUSIONS: These findings show that phosphorylation at Ser(167) of ER predicts for likelihood of response of ER positive breast cancer patients to endocrine therapies. PMID- 17908968 TI - Short telomeres: a novel potential predictor of relapse in Ewing sarcoma. AB - PURPOSE: Despite advances in therapy, >50% of patients with Ewing sarcoma will relapse. The current prognostic factors are not optimal for risk prediction. Studies have shown that telomere length could predict outcome in different malignancies. Our aim was to evaluate whether telomere length could be a better prognostic factor in Ewing sarcoma and correlate the results with clinical variables, outcome, and chromosomal instability. EXPERIMENTAL DESIGN: Telomere length was determined in the primary tumor and peripheral blood of 32 patients with Ewing sarcoma. Chromosomal instability was evaluated by combining classical cytogenetics, comparative genomic hybridization and random aneuploidy. Telomere length was correlated to clinical variables, chromosomal instability, and outcome. RESULTS: In 75% of the tumors, changes in telomere length, when compared with the corresponding peripheral blood lymphocytes, were noted. The majority of changes consisted of a reduction in telomere length. Patients harboring shorter telomeres had a significantly adverse outcome (P = 0.015). Chromosomal instability was identified in 65% of tumors, significantly correlating with short telomeres (P = 0.0094). Using multivariate analysis, telomere length remained the only significant prognostic variable (P = 0.034). Patients with short telomeres had a 5.3-fold risk of relapse as compared to those with unchanged or longer telomeres. CONCLUSION: We have shown that tumors with telomere length reduction result in genomic instability. In addition, telomere length reduction was the only significant predictor of outcome. We suggest that reduction of telomere length in tumor cells at diagnosis could serve as a prognostic marker in Ewing sarcoma. PMID- 17908969 TI - A high tumor-associated macrophage content predicts favorable outcome in follicular lymphoma patients treated with rituximab and cyclophosphamide doxorubicin-vincristine-prednisone. AB - PURPOSE: Tumor-associated macrophage (TAM) content predicts survival in follicular lymphoma (FL) patients treated with chemotherapy. The aim of this study was to determine how combination of rituximab with chemotherapy influences TAM-associated clinical outcome. EXPERIMENTAL DESIGN: Expression of a macrophage marker, CD68, was determined immunohistochemically from FL samples of 96 patients treated with rituximab and cyclophosphamide-Adriamycin-vincristine-prednisone regimen. Of them, 71 received therapy at diagnosis and 25 at relapse. Neutrophil and CD3+ lymphocyte counts were also measured. The median follow-up time for the cohort was 54 months. Fourty-five patients previously treated with chemotherapy served as a control group. RESULTS: Consistent with previous studies, high TAM amount was associated with adverse outcome in chemotherapy-treated patients (P = 0.026). In contrast, after rituximab and cyclophosphamide-doxorubicin-vincristine prednisone regimen, high TAM content correlated with longer survival rates. According to Kaplan Meier estimates, the median progression free survival (PFS) was not reached for patients with high TAM content compared with 45 months for patients with low TAM scores (P = 0.006). A trend toward a better overall survival (OS) at 5 years was also observed for patients with high TAM content (OS, 97% versus 90%, P = 0.116). The positive prognostic value of TAMs was seen both for the patients treated at diagnosis and at relapse. In multivariate analyses, TAM content remained an independent prognostic factor for OS and PFS. Neutrophil and CD3+ lymphocyte counts did not correlate with outcome. CONCLUSIONS: The data suggest that high TAM score is associated with a favorable prognosis in FL patients treated with immunochemotherapy. PMID- 17908970 TI - The AA genotype of the regulatory BCL2 promoter polymorphism ( 938C>A) is associated with a favorable outcome in lymph node negative invasive breast cancer patients. AB - PURPOSE: Expression of the antiapoptotic and antiproliferative protein Bcl-2 has been repeatedly shown to be associated with better clinical outcome in breast cancer. We recently showed a novel regulatory (-938C>A) single-nucleotide polymorphism (SNP) in the inhibitory P2 BCL2 gene promoter generating significantly different BCL2 promoter activities. EXPERIMENTAL DESIGN: Paraffin embedded neoplastic and nonneoplastic tissues from 274 patients (161 still alive after a follow-up period of at least 80 months) with primary unilateral invasive breast carcinoma were investigated. Bcl-2 expression of tumor cells was shown by immunohistochemistry; nonneoplastic tissues were used for genotyping. Both the Bcl-2 expression and the (-938C>A) genotypes were correlated with the patients' survival. RESULTS: Kaplan-Meier curves revealed a significant association of the AA genotype with increased survival (P = 0.030) in lymph node-negative breast cancer patients, whereas no genotype effect could be observed in lymph node positive cases. Ten-year survival rates were 88.6% for the AA genotype, 78.4% for the AC genotype, and 65.8% for the CC genotype. Multivariable Cox regression identified the BCL2 (-938CC) genotype as an independent prognostic factor for cancer-related death in lymph node-negative breast carcinoma patients (hazard ratio, 3.59; P = 0.032). Immunohistochemical Bcl-2 expression was significantly associated with the clinical outcome of lymph node-positive but not of lymph node negative breast cancer patients. In lymph node-negative cases, the (-938C>A) SNP was both significantly related with the immunohistochemically determined level of Bcl-2 expression (P = 0.044) and the survival of patients with Bcl-2-expressing carcinomas (P = 0.006). CONCLUSIONS: These results suggest the (-938C>A) polymorphism as a survival prognosticator as well as indicator of a high-risk group within patients with lymph node-negative breast cancer. PMID- 17908971 TI - p66 Shc tumor levels show a strong prognostic correlation with disease outcome in stage IIA colon cancer. AB - PURPOSE: Most stage IIA colon cancer patients receive no adjuvant therapy despite an estimated 15% risk of disease-related death within 5 years of resection. Prognostication of disease outcome would benefit the clinician by categorizing patients with stage IIA disease by risk. The abundance of the signal transduction proteins p66 Shc and tyrosine-phosphorylated (PY)-Shc in tumor cells is a prognostic indicator of disease outcome in breast cancer, suggesting that Shc analysis may provide prognostic information in stage IIA colon cancer. EXPERIMENTAL DESIGN: Immunohistochemical staining of p66 Shc and PY-Shc was examined in resection specimens from 240 chemotherapy-naive patients with stage IIA (T(3)N(0)M(0)) colon cancer from two independent (130 and 110 cases, respectively) retrospective cohorts. Staining was scored on a 0 to 5 scale and correlated with relapse-free survival and disease-specific survival in a multivariate analysis to obtain hazard ratios (HR) for both outcomes. RESULTS: In a pooled analysis of both cohorts, p66 Shc score was a significant prognostic indicator of relapse-free survival (full-range HR, 13.0; P = 0.012) and disease specific survival (full-range HR, 36.6; P = 0.004) when analyzed as a continuous variable in a multivariate Cox proportional hazards model stratified by study site and adjusted for age, sex, grade, and lymphovascular involvement. PY-Shc in this multivariate Cox model, however, did not achieve statistical significance for either outcome. CONCLUSIONS: Measuring p66 Shc tumor levels provides a unique and simple tool for stratifying stage IIA colon cancer patients by risk of recurrence and disease-specific death and may assist in determining treatment strategies for these patients. PMID- 17908972 TI - Measurement of cyclin E genomic copy number and strand length in cell-free DNA distinguish malignant versus benign effusions. AB - PURPOSE: Previous studies have shown that the concentration of cell-free DNA was higher and its strand length longer in body fluids obtained from patients with cancer as compared to patients with benign diseases. We hypothesized that analysis of both DNA copy number and strand length of cell-free DNA from an amplified chromosomal region could improve the diagnosis of malignant diseases in body fluids. EXPERIMENTAL DESIGN: To test this hypothesis, we used ovarian cancer effusion as an example and applied a quantitative real-time PCR to measure the relative copy number and strand length of DNA fragments from one of the most frequently amplified genes, cyclin E, in ovarian serous carcinomas. RESULTS: As compared with nonamplified chromosomal loci, including beta-actin, p53, 2p24.1, and 4p15.31, measurement of cyclin E DNA copy number (100 bp) had the best performance in distinguishing malignant (n = 88) from benign (n = 70) effusions after normalization to effusion volume or Line-1 DNA with areas under the receiver operating characteristics curve (AUC) of 0.832 and 0.847, respectively. Different DNA lengths of the cyclin E locus were further analyzed and we found that the AUC was highest by measuring the 400-bp cyclin E locus (AUC = 0.896). The AUC was improved to 0.936 when it was combined with the length integrity index as defined by the relative abundance of 400 bp cyclin E to 100 bp p53 loci. Cyclin E real-time PCR assay had a higher sensitivity (95.6%) than routine cytology examination (73.9%) and was able to diagnose false-negative cytology cases in this study. CONCLUSIONS: The above findings indicate that measurement of the DNA copy number and strand length of the cyclin E locus is a useful cancer diagnostic tool. PMID- 17908973 TI - Caspase-3 activity predicts local recurrence in rectal cancer. AB - PURPOSE: Radiotherapy followed by total mesorectal excision surgery has been shown to significantly reduce local recurrence rates in rectal cancer patients. Radiotherapy, however, is associated with considerable morbidity. The present study evaluated the use of biochemical detection of enzymatic caspase-3 activity as preoperative marker for apoptosis to preselect patients that are unlikely to develop a local recurrence to spare these patients from overtreatment and the negative side effects of radiotherapy. EXPERIMENTAL DESIGN: Nonirradiated freshly frozen tissue samples from 117 stage III rectal cancer patients were collected from a randomized clinical trial that evaluated preoperative radiotherapy in total mesorectal excision surgery. Additional frozen archival tissues from 47 preoperative biopsies and corresponding resected colorectal tumors were collected. Level of apoptosis was determined by measuring the enzymatic activity of caspase-3 in a biochemical assay. RESULTS: In tumor tissue, caspase-3 activity lower than the median was predictive of 5-year local recurrence (hazard ratio, 7.4; 95% confidence interval, 1.7-32.8; P = 0.008), which was unaffected by adjustment for type of resection, tumor location, and T status (adjusted hazard ratio, 7.5; 95% confidence interval, 1.7-34.1; P = 0.009). Caspase-3 activity in preoperative biopsies was significantly correlated with caspase-3 activity in corresponding resected tumors (r = 0.56; P < 0.0001). CONCLUSION: Detection of tumor apoptosis levels by measuring caspase-3 activity, for which a preoperative biopsy can be used, accurately predicted local recurrence in rectal cancer patients. These findings indicate that caspase-3 activity is an important denominator of local recurrence and should be evaluated prospectively to be added to the criteria to select rectal cancer patients in which radiotherapy is redundant. PMID- 17908974 TI - Platelet-derived growth factor receptor inhibition and chemotherapy for castration-resistant prostate cancer with bone metastases. AB - PURPOSE: To further assess preclinical and early clinical evidence that imatinib mesylate, a platelet-derived growth factor receptor (PDGFR) inhibitor, modulates taxane activity in prostate cancer and bone metastases, a randomized study was conducted. EXPERIMENTAL DESIGN: Men with progressive castration-resistant prostate cancer with bone metastases (n = 144) were planned for equal randomization to i.v. 30 mg/m(2) docetaxel on days 1, 8, 15, and 22 every 42 days with 600 mg imatinib daily or placebo, for an improvement in median progression free survival from 4.5 to 7.5 months (two-sided alpha = 0.05 and beta = 0.20). Secondary end points included differential toxicity and bone turnover markers, tumor phosphorylated PDGFR (p-PDGFR) expression, and modulation of p-PDGFR in peripheral blood leukocytes. RESULTS: Accrual was halted early because of adverse gastrointestinal events. Among 116 evaluable men (57 docetaxel + imatinib; 59 docetaxel + placebo), respective median times to progression were 4.2 months (95% confidence interval, 3.1-7.5) and 4.2 months (95% confidence interval, 3.0-6.8; P = 0.58, log-rank test). Excess grade 3 toxicities (n = 23) in the docetaxel + imatinib group were principally fatigue and gastrointestinal. Tumor p-PDGFR expression was observed in 12 of 14 (86%) evaluable bone specimens. In peripheral blood leukocytes, p-PDGFR reduction was more likely in docetaxel + imatinib treated patients compared with docetaxel + placebo (P < 0.0001), as were reductions in urine N-telopeptides (P = 0.004) but not serum bone-specific alkaline phosphatase (P = 0.099). CONCLUSIONS: These clinical and translational results question the value of PDGFR inhibition with taxane chemotherapy in prostate cancer bone metastases and are at variance with the preclinical studies. This discordance requires explanation. PMID- 17908975 TI - Molecular alterations in prostate carcinomas that associate with in vivo exposure to chemotherapy: identification of a cytoprotective mechanism involving growth differentiation factor 15. AB - PURPOSE: To identify molecular alterations associating with in vivo exposure of prostate carcinoma to chemotherapy and assess functional roles modulating tumor response and resistance. EXPERIMENTAL DESIGN: Patients with high-risk localized prostate cancer (tumor-node-metastasis >or= T(2b) or prostate-specific antigen >or= 15 ng/mL or Gleason glade >or= 4+3) were enrolled into a phase II clinical trial of neoadjuvant chemotherapy with docetaxel and mitoxantrone followed by prostatectomy. Pretreatment prostate tissue was acquired by needle biopsy and posttreatment tissue was acquired by prostatectomy. Prostate epithelium was captured by microdissection, and transcript levels were quantitated by cDNA microarray hybridization. Gene expression changes associated with chemotherapy were determined by a random variance t test. Several were verified by quantitative reverse transcription PCR. In vitro analyses determining the influence of growth differentiation factor 15 (GDF15) on chemotherapy resistance were done. RESULTS: Gene expression changes after chemotherapy were measured in 31 patients who completed four cycles of neoadjuvant chemotherapy. After excluding genes shown previously to be influenced by the radical prostatectomy procedure, we identified 51 genes with significant transcript level alterations following chemotherapy. This group included several cytokines, including GDF15, chemokine (C-X-C motif) ligand 10, and interleukin receptor 1beta. Overexpression of GDF15 or exposure of prostate cancer cell lines to exogenous recombinant GDF15 conferred resistance to docetaxel and mitoxantrone. CONCLUSIONS: Consistent molecular alterations were identified in prostate cancer cells exposed to docetaxel and mitoxantrone chemotherapy. These alterations include transcripts encoding cytokines known to be regulated through the nuclear factor-kappaB pathway. Chemotherapy-induced cytokines and growth factors, such as GDF15, contribute to tumor cell therapy resistance and may serve as targets to improve responses. PMID- 17908976 TI - Phase I dose escalation study of the anti insulin-like growth factor-I receptor monoclonal antibody CP-751,871 in patients with refractory solid tumors. AB - PURPOSE: This phase I study was undertaken to define the maximum tolerated dose, safety, and pharmacokinetic profile of CP-751,871. EXPERIMENTAL DESIGN: Using a rapid dose escalation design, patients with advanced nonhematologic malignancies were treated with CP-751,871 in four dose escalation cohorts. CP-751,871 was administered i.v. on day 1 of each 21-day cycle. Pharmacokinetic evaluation was done in all treatment cohorts during cycles 1 and 4. RESULTS: Twenty-four patients received 110 cycles at four dose levels. The maximum tolerated dose exceeded the maximal feasible dose of 20 mg/kg and, thus, was not identified. Treatment-related toxicities were generally mild. The most common adverse events were hyperglycemia, anorexia, nausea, elevated aspartate aminotransferase, elevated gamma-glutamyltransferase, diarrhea, hyperuracemia, and fatigue. At 20 mg/kg, 10 of 15 patients experienced stability of disease. Two of these patients experienced long-term stability. There were no objective responses. Pharmacokinetic analysis revealed a dose-dependent increase in CP-751,871 exposure and approximately 2-fold accumulation on repeated dosing in 21-day cycles. Plasma concentrations of CP-751,871 attained were several log-fold greater than the biologically active concentration. Treatment with CP-751,871 increased serum insulin and human growth hormone levels, with modest increases in serum glucose levels. CONCLUSIONS: CP-751,871 has a favorable safety profile and was well tolerated when given in continuous cycles. At the maximal feasible dose of 20 mg/kg, there was a moderate accumulation in plasma exposure, and most of the treated patients experienced stability of disease. PMID- 17908977 TI - Phase I dose-finding study of weekly docetaxel followed by flavopiridol for patients with advanced solid tumors. AB - PURPOSE: Flavopiridol is a cyclin-dependent kinase inhibitor that enhances docetaxel-induced apoptosis in a sequence-specific manner. In vivo, docetaxel must precede flavopiridol by at least 4 h to induce this effect. We conducted a phase I trial of weekly, sequential docetaxel followed 4 h later by flavopiridol in patients with advanced solid tumors. EXPERIMENTAL DESIGN: Docetaxel at a fixed dose of 35 mg/m2 was administered over 30 min, followed 4 h later by escalating doses of flavopiridol, ranging from 20 to 80 mg/m2 in successive cohorts, administered weekly over 1 h. This schedule was repeated for 3 weeks of each 4 week cycle. RESULTS: Twenty-seven evaluable patients were enrolled. The combination was well tolerated, with one dose-limiting toxicity occurring at flavopiridol 70 mg/m2 (grade 3 mucositis) and one dose-limiting toxicity at 80 mg/m2 (grade 4 neutropenia). We observed 1 complete response in a patient with pancreatic carcinoma and 4 partial responses in pancreatic (1), breast (2), and ovarian (1) cancer patients. Stable disease was seen in 10 patients. Pharmacokinetic studies showed Cmax ranging from 1.49 +/- 0.69 micromol/L (flavopiridol 20 mg/m2) to 4.54 +/- 0.08 micromol/L (flavopiridol 60 mg/m2) in cycle 1. CONCLUSIONS: Treatment with weekly, sequential docetaxel followed by flavopiridol is an effective and safe regimen at all flavopiridol dose levels. The pharmacokinetic data indicate that concentrations of flavopiridol that enhance the effects of docetaxel both in vitro and in vivo can be achieved. Clinical activity is encouraging, even in patients who have received a prior taxane and in patients with gemcitabine-refractory metastatic pancreatic cancer. PMID- 17908978 TI - Adjuvant adenovirus-mediated delivery of herpes simplex virus thymidine kinase administration improves outcome of liver transplantation in patients with advanced hepatocellular carcinoma. AB - PURPOSE: Previous poor results of liver transplantation (LT) have been confirmed in patients with advanced hepatocellular carcinoma (HCC). Adenovirus-mediated delivery of herpes simplex virus thymidine kinase (ADV-TK) therapy is an established adjuvant treatment in cancer, and we evaluated its potential as an adjuvant treatment for HCC patients who underwent LT. EXPERIMENTAL DESIGN: Forty five HCC patients with tumors >5 cm in diameter participated in the study over a follow-up period of 50 months. Among these patients, 22 received LT only, and 23 received LT combined with ADV-TK therapy. All HCC patients enrolled in this study had tumors >5 cm in diameter and no metastasis in lungs or bones was detected by computed tomography or magnetic resonance imaging scans. RESULTS: The recurrence free survival and the overall survival in the LT plus ADV-TK therapy group were 43.5% and 69.6%, respectively, at 3 years; both values were significantly higher than those in the LT-only group (9.1% and 19.9%, respectively). In the nonvascular invasion subgroup, overall survival was 100% and recurrence-free survival was 83.3% in the patients receiving LT plus ADV-TK, significantly higher than the patients receiving LT only. CONCLUSIONS: HCC patients with no vascular invasion could be selected for LT followed by adjuvant ADV-TK therapy, regardless of intrahepatic huge or diffuse tumor. We propose that the current criteria for LT based on tumor size may be expanded if accompanied by ADV-TK therapy due to improved prognosis. PMID- 17908979 TI - Phase I trial of poly-L-glutamate camptothecin (CT-2106) administered weekly in patients with advanced solid malignancies. AB - PURPOSE: CT-2106 is a 20(S)-camptothecin poly-L-glutamate conjugate. This linkage stabilizes the active lactone form of camptothecin and enhances aqueous solubility. In addition, poly-L-glutamate is postulated to increase tumor delivery of the active compound through enhanced permeability and retention effect in tumor. We studied a weekly schedule of CT-2106 in patients with refractory solid tumor malignancies. EXPERIMENTAL DESIGN: CT-2106 was infused (10 min i.v. infusion) on days 1, 8, and 15 of each 28-day cycle. Plasma and urine were analyzed for total and unconjugated camptothecin by high-performance liquid chromatography equipped with a fluorescence detector. Toxicity and response assessments were done with Common Toxicity Criteria for Adverse Events version 3 and Response Evaluation Criteria in Solid Tumors, respectively. RESULTS: Twenty six patients were enrolled. Median age was 58 years (range, 36-83) and median number of doses was 6 (range, 1-9). The most frequent tumor type (50%) was melanoma. Dose limiting toxicities were thrombocytopenia and fatigue. A weekly dose of 25 mg/m2 given every 3 of 4 weeks was the maximum tolerated dose. The majority of grade 3 and 4 toxicities were hematologic. The pharmacokinetic profile of conjugated and unconjugated camptothecin showed a polyexponential decline with similar terminal half life (t1/2 range was 44-63 and 31-48 h for conjugated and unconjugated, respectively). Pharmacokinetics of conjugated and unconjugated camptothecin were dose and time independent in the tested dose range. Urinary excretion of conjugated and unconjugated camptothecin accounted for about 30% and 4% of the administered dose, respectively. CONCLUSIONS: CT-2106 has a more manageable toxicity profile compared with unconjugated camptothecin. The maximum tolerated dose is 25 mg/m2 weekly given 3 of 4 weeks. This compound results in prolonged release of unconjugated camptothecin. PMID- 17908980 TI - Inflammatory breast cancer as a model disease to study tumor angiogenesis: results of a phase IB trial of combination SU5416 and doxorubicin. AB - PURPOSE: We used inflammatory breast cancer (IBC) as a model disease to investigate biological changes associated with an antiangiogenesis agent, SU5416, combined with doxorubicin. EXPERIMENTAL DESIGN: Patients with stage IIIB or IV IBC were treated neoadjuvantly with the combination of SU5416 and doxorubicin for induction therapy. The dose of SU5416 (administered on days 1 and 4, every 3 weeks) and doxorubicin (administered on day 1 every 3 weeks) were escalated in cohorts of three patients starting at 110 and 60 mg/m2, respectively, for a total of five cycles leading up to mastectomy. Patients underwent serial assessment (pharmacokinetic sampling, biopsy of breast, tumor blood flow dynamic contrast enhanced magnetic resonance imaging, plasma angiogenesis, and endothelial cell damage markers) prior to treatment, at the end of cycles no. 2 and no. 5, and after mastectomy. RESULTS: Eighteen patients were enrolled; neutropenia was dose limiting, and overall median survival was not reached (50 months of study follow up). Four patients (22%) experienced congestive heart failure, which resolved and were likely attributable to a smaller volume of distribution and higher Cmax of doxorubicin in combination with SU5416. We did observe a significant decline in tumor blood flow using Kep calculated by Brix (pretreatment versus post-cycle no. 5; P = 0.033), trend for a decline in tumor microvessel density after treatment, and low baseline levels of soluble intracellular adhesion molecule were associated with improved event-free survival. CONCLUSIONS: This study showed evidence of an unfavorable cardiac interaction between SU5416 and doxorubicin, which prohibits further investigation of this combination. However, this study supports the importance of using IBC as a model for investigating angiogenesis inhibitors. PMID- 17908981 TI - A phase II study of gefitinib monotherapy in advanced esophageal adenocarcinoma: evidence of gene expression, cellular, and clinical response. AB - PURPOSE: At presentation, most cases of adenocarcinoma of the esophagus (ACE) are inoperable. Although chemotherapy can prolong survival, patients eventually die as a result of refractory disease. Epidermal growth factor receptor (EGFR) is almost universally expressed in ACE and is a negative prognostic factor. EXPERIMENTAL DESIGN: This open-label, two-center, noncomparative, two-part phase II trial assessed the EGFR tyrosine kinase inhibitor gefitinib (500 mg/d) in patients with advanced, inoperable ACE. The primary end point was tumor response. The effect of EGFR inhibition was also evaluated by gene expression analysis of tumor biopsies taken before gefitinib treatment and 28 days after. RESULTS: Twenty-seven patients were recruited and evaluable for tumor response and safety. Three patients had a partial response and seven had stable disease, giving a disease control rate (partial response + stable disease) of 37%. Drug-related adverse events were generally mild: diarrhea in 19 (grade 3 in three) and rash in 19 (grade 3 in five) patients, and there were no grade 4 drug-related adverse events. Microarray experiments on tumor biopsies showed that gefitinib also down regulated oncogenes associated with tumor progression. Ki67 (a marker of tumor growth) expression decreased in five of seven biopsies taken before and after treatment. CONCLUSION: Gefitinib (500 mg/d) is an active and generally well tolerated treatment for ACE. Studies on endoscopic biopsies are feasible and indicate that gefitinib inhibits both gene expression and cellular biology at 500 mg/d, and these may provide surrogate end points for predictive biomarkers. Further trials of gefitinib are warranted, particularly as patient response seems to be durable and current second-line chemotherapy options have no proven ability to prolong life. PMID- 17908982 TI - Phase I and pharmacokinetic study of imatinib mesylate (Gleevec) and gemcitabine in patients with refractory solid tumors. AB - PURPOSE: Preclinical data shows improvements in response for the combination of imatinib mesylate (IM, Gleevec) and gemcitabine (GEM) therapy compared with GEM alone. Our goals were to determine the maximum tolerated dose of GEM and IM in combination, the pharmacokinetics of GEM in the absence and in the presence of IM, and IM pharmacokinetics in this combination. PATIENTS AND METHODS: Patients with refractory malignancy, intact intestinal absorption, measurable/evaluable disease, adequate organ function, Eastern Cooperative Oncology Group PS 0-2, and signed informed consent were eligible. Initially, treatment consisted of 600 mg/m2 of GEM (10 mg/m2/min) on days 1, 8, and 15, and 300 mg of IM daily every 28 days. Due to excessive toxicity, the schedule was altered to IM on days 1 to 5 and 8 to 12, and GEM on days 3 and 10 every 21 days. Two final cohorts received IM on days 1 to 5, 8 to 12, and 15 to 19. RESULTS: Fifty-four patients were treated. IM and GEM given daily at 500 to 600 mg/m2 on days 1, 8, and 15 produced frequent dose-limiting toxicities. With the modified scheduling, GEM given at 1,500 mg/m2/150 min was deliverable, along with 400 mg of IM, without dose limiting toxicities. Three partial (laryngeal, renal, and mesothelioma) and two minor (renal and pancreatic) responses were noted at GEM doses of 450 to 1,500 mg/m2. Stable disease >24 weeks was seen in 17 patients. CA19-9 in 7 of 10 patients with pancreatic cancer was reduced by approximately 90%. IM did not significantly alter GEM pharmacokinetics. CONCLUSION: The addition of intermittently dosed IM to GEM at low to full dose was associated with broad antitumor activity and little increase in toxicity. PMID- 17908984 TI - Characterization of a putative ovarian oncogene, elongation factor 1alpha, isolated by panning a synthetic phage display single-chain variable fragment library with cultured human ovarian cancer cells. AB - PURPOSE: In an effort to identify cell surface targets and single short-chain antibody (scFv) for ovarian cancer therapy, we used a phage display approach to isolate an antibody with high reactivity against ovarian cancer. EXPERIMENTAL DESIGN: A phage scFv library was subjected to panning against human SK-OV-3 ovarian cancer cells. A clone with high reactivity was selected and tested in immunoperoxidase staining on a panel of normal tissues and ovarian carcinoma. Using immunoprecipitation, a differentially expressed band was analyzed by mass spectrometry. The antigen subclass was characterized with reverse transcription PCR on cDNA library of normal tissues, and 91 ovarian cancer specimens, and correlated with clinicohistopathologic characteristics. RESULTS: Ninety-six individual scFv clones were screened in ELISA following panning. scFv F7 revealed high reactivity with ovarian cancer cell lines and showed intense staining of 15 fresh ovarian cancer specimens and no staining of a panel of normal tissues. A 40 kDa protein was identified to be translation elongation factor 1alpha1 (EEF1A1; P < 0.05). The expression of EEF1A2, a highly homologous and functionally similar oncogene, was found to be restricted only to the normal tissues of the heart, brain, and skeletal muscle. Aberrant EEF1A2 mRNA expression was found in 21 of 91 (23%) of ovarian cancer specimens and significantly correlated with increased likelihood of recurrence (P = 0.021). CONCLUSIONS: scFv F7 may represent an ovarian cancer-specific antibody against translation EEF1A family of translational factors. We propose that EEF1A2 may be a useful target for therapy of human ovarian cancer. PMID- 17908983 TI - Preclinical testing of clinically applicable strategies for overcoming trastuzumab resistance caused by PTEN deficiency. AB - PURPOSE: We have previously shown that PTEN loss confers trastuzumab resistance in ErbB2-overexpressing breast cancer using cell culture, xenograft models, and patient samples. This is a critical clinical problem because trastuzumab is used in a variety of therapeutic regimens, and at the current time, there are no established clinical strategies to overcome trastuzumab resistance. Here, we did preclinical studies on the efficacy of clinically applicable inhibitors of the Akt/mammalian target of rapamycin (mTOR) pathway to restore trastuzumab sensitivity to PTEN-deficient cells. EXPERIMENTAL DESIGN: Cell culture and xenograft models were used to test a panel of clinically applicable, small molecule inhibitors of the Akt/mTOR signal transduction pathway, a critical pathway downstream of ErbB2, and identify compounds with the ability to restore trastuzumab sensitivity to PTEN-deficient cells. RESULTS: When trastuzumab was combined with the Akt inhibitor triciribine, breast cancer cell growth was inhibited and apoptosis was induced. In a xenograft model, combination therapy with trastuzumab and triciribine dramatically inhibited tumor growth. The combination of trastuzumab and the mTOR inhibitor RAD001 also slowed breast cancer cell growth in vitro and in vivo. CONCLUSIONS: Combining trastuzumab with inhibitors of the Akt/mTOR pathway is a clinically applicable strategy and combinations of trastuzumab with triciribine or RAD001 are promising regimens for rescue of trastuzumab resistance caused by PTEN loss. PMID- 17908985 TI - Herpes simplex virus Us3(-) mutant as oncolytic strategy and synergizes with phosphatidylinositol 3-kinase-Akt targeting molecular therapeutics. AB - PURPOSE: Oncolytic herpes simplex virus (HSV) vectors have shown safety in clinical trials, but efficacy remains unsatisfactory. Novel HSV vectors that possess tumor selectivity with enhanced potency are therefore needed. The gene product of HSV Us3 protects virus-infected cells from apoptosis, a cellular pathway frequently dysfunctional in tumors. We hypothesized that Us3 mutants, whose replication would be inhibited by apoptosis in normal cells, would be selective for tumor cells. EXPERIMENTAL DESIGN: HSV mutants G207 (ribonucleotide reductase-/gamma34.5-), R7041 (Us3-), and R7306 (Us3 revertant) were tested in normal and tumor cells for viral replication, antitumoral potency, apoptosis induction, and Akt activation. Safety and biodistribution after systemic administration and antitumoral efficacy after intratumoral (i.t.) or i.v. administration were examined. RESULTS: Us3 deletion results in up to 3-log replication inhibition in normal cells, which correlates with enhanced apoptosis induction. In contrast, R7041 replicates very well in tumor cells, showing 1 to 2 log greater yield than G207. In vivo, R7041 shows no signs of toxicity after systemic delivery in both immunocompetent and immunodeficient mice and shows preferential and prolonged replication in tumors compared with normal tissues. R7041 displays significant antitumoral efficacy after i.t. or i.v. administration. An additional feature of Us3 mutants is enhanced Akt activation compared with wild-type infection, which sensitizes cells to phosphatidylinositol 3-kinase-Akt inhibitors (LY294002, Akt inhibitor IV), shown by synergistic antitumoral activity in vitro and enhanced efficacy in vivo. CONCLUSIONS: Us3 deletion confers enhanced tumor selectivity and antitumoral potency on herpes simplex virus-1 and provides for a novel mechanism of combination therapy with phosphatidylinositol 3-kinase-Akt-targeting molecular therapeutics. PMID- 17908986 TI - Neutralizing B-cell activating factor antibody improves survival and inhibits osteoclastogenesis in a severe combined immunodeficient human multiple myeloma model. AB - PURPOSE: B-cell-activating factor (BAFF) is a tumor necrosis factor superfamily member critical for the maintenance and homeostasis of normal B-cell development. It has been implicated in conferring a survival advantage to B-cell malignancies, including multiple myeloma (MM). EXPERIMENTAL DESIGN: Here, we validate the role of BAFF in the in vivo pathogenesis of MM examining BAFF and its receptors in the context of patient MM cells and show activity of anti-BAFF antibody in a severe combined immunodeficient model of human MM. RESULTS: Gene microarrays and flow cytometry studies showed increased transcripts and the presence of all three receptors for BAFF in CD138+ patient MM cells, as well as an increase in plasma BAFF levels in 51 MM patients. Functional studies show that recombinant BAFF protects MM cells against dexamethasone-induced apoptosis accompanied by an increase in survival proteins belonging to the BCL family. These in vitro studies led to the evaluation of a clinical grade-neutralizing antibody to BAFF in a severe combined immunodeficient human MM model. Anti-BAFF-treated animals showed decreased soluble human interleukin 6 receptor levels, a surrogate marker of viable tumor, suggesting direct anti-MM activity. This translated into a survival advantage of 16 days (P < 0.05), a decrease in tartrate-resistant acid phosphatase-positive osteoclasts, and a reduction in radiologically evident lytic lesions in anti-BAFF-treated animals. CONCLUSIONS: Our data show a role for BAFF as a survival factor in MM. Importantly, the in vivo antitumor activity of neutralizing anti-BAFF antibody provide the preclinical rationale for its evaluation in the treatment of MM. PMID- 17908987 TI - Inhibition of Jun NH2-terminal kinases suppresses the growth of experimental head and neck squamous cell carcinoma. AB - PURPOSE: This study was carried out to investigate whether c-Jun NH2-terminal kinases (JNK) are potential targets for treating head and neck squamous cell carcinoma (HNSCC). EXPERIMENTAL DESIGN: JNK activity was first evaluated in 20 paired samples of human HNSCC. The antitumor activity of SP600125, a reversible nonselective ATP-competitive inhibitor of JNKs, was then investigated both in an HNSCC xenograft model and in vitro using immunohistochemistry, immunoblotting, enzyme immunoassay, flow cytometry, and a Matrigel assay of capillary tube formation. Complementary studies were carried out using small interfering RNA to JNK1/2. RESULTS: JNK activity was increased in human HNSCC compared with normal appearing epithelium. Treatment of mice bearing HNSCC xenografts with SP600125 resulted in >60% inhibition of tumor growth relative to vehicle-treated animals. Inhibition of tumor growth was associated with significant reductions in both cell proliferation and microvessel density. SP600125 inhibited tumor cell proliferation by causing delays in both the S and G2-M phases of the cell cycle. Inhibition of angiogenesis seemed to reflect effects on both tumor and endothelial cells. The JNK inhibitor suppressed the production of vascular endothelial growth factor and interleukin-8 by tumor cells and also inhibited endothelial cell proliferation and capillary tube formation. Reduced amounts and phosphorylation of epidermal growth factor receptor were found in tumor cells after treatment with SP600125. Small interfering RNA-mediated suppression of JNK1/2 led to reduced tumor cell proliferation and decreased levels of epidermal growth factor receptor, vascular endothelial growth factor, and interleukin-8. CONCLUSIONS: JNK activity is commonly increased in HNSCC. Our preclinical results provide a rationale for evaluating JNK inhibition as an approach to treating HNSCC. PMID- 17908988 TI - The therapeutic efficacy of anti vascular endothelial growth factor antibody, bevacizumab, and pemetrexed against orthotopically implanted human pleural mesothelioma cells in severe combined immunodeficient mice. AB - PURPOSE: Malignant pleural mesothelioma (MPM) is an aggressive malignancy, which has a poor prognosis with a median survival of less than 1 year. The vascular endothelial growth factor (VEGF) has been reported to be an ideal therapeutic target, and a multitargeted antifolate, pemetrexed, has been clinically used for the treatment of MPM. EXPERIMENTAL DESIGN: We examined the therapeutic efficacy of the antihuman VEGF neutralizing antibody, bevacizumab, in combination with pemetrexed against two different human MPM cells, EHMES-10 and MSTO-211H, orthotopically inoculated into severe combined immunodeficient mice. RESULTS: Bevacizumab inhibited a VEGF-induced proliferation of the human endothelial cells in a dose-dependent manner, but it had no effect on the proliferation of the two MPM cell lines in vitro. The orthotopically inoculated EHMES-10 cells (VEGF high expressing) produced thoracic tumors and a large volume of bloody pleural effusion, whereas the MSTO-211H cells (VEGF low expressing) produced thoracic tumors and a small volume of bloody effusions. Treatment with bevacizumab effectively inhibited the production of thoracic tumors and dramatically prevented the production of pleural effusion by the EHMES-10 cells but not the MSTO-211H cells. Treatment with bevacizumab reduced the number of enlarged tumor associated vessels and proliferating tumor cells. Moreover, treatment with bevacizumab in combination with pemetrexed more effectively suppressed the formation of the pleural effusion and prolonged the survival compared with the control and monotherapy in the EHMES-10 cell-bearing severe combined immunodeficient mice. CONCLUSIONS: These results suggest that the combined use of bevacizumab and pemetrexed may therefore be promising for controlling the progression of MPM highly expressing VEGF. PMID- 17908989 TI - Anthrax lethal toxin inhibits growth of and vascular endothelial growth factor release from endothelial cells expressing the human herpes virus 8 viral G protein coupled receptor. AB - PURPOSE: In this study, we tested the hypothesis that inhibition of mitogen activated protein kinase kinases (MKK) inhibits tumor growth by acting on angiogenic signaling and by extension may form the basis of an effective strategy for treatment of Kaposi's sarcoma. EXPERIMENTAL DESIGN: Murine endothelial cells expressing the human herpes virus 8 G protein-coupled receptor (vGPCR-SVEC) were treated with anthrax lethal toxin (LeTx). LeTx is a binary toxin ordinarily secreted by Bacillus anthracis and is composed of two proteins: protective antigen (the binding moiety) and lethal factor (the active moiety). Lethal factor is a protease that cleaves and inactivates MKKs. RESULTS: In vitro, treatment of vGPCR-SVEC with LeTx inhibited MKK signaling, moderately inhibited cell proliferation, and blocked the ability of these cells to form colonies in soft agar. Treatment with LeTx also blocked the ability of these cells to release several angioproliferative cytokines, notably vascular endothelial growth factor (VEGF). In contrast, inhibition of mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2 with U0126 caused a substantial inhibition of proliferation but only modestly inhibited VEGF release. In xenograft models, i.v. injection of LeTx caused reduced tumor growth characterized immunohistochemically by inhibition of MKK signaling, decreased rates of proliferation, and reduced levels of VEGF and VEGF receptor 2, with a corresponding decrease in vascular density. CONCLUSIONS: These data support a role for MKK signaling in tumor growth and vascularization and are consistent with the hypothesis that inhibition of MKK signaling by LeTx or a similar agent may be an effective strategy for the treatment of Kaposi's sarcoma as well as other vascular tumors. PMID- 17908990 TI - Imaging and modulating antisense microdistribution in solid human xenograft tumor models. AB - PURPOSE: The tumor microenvironment is complex and heterogeneous, populated by tortuous irregular vasculature, hypoxic cells, and necrotic regions. These factors can all contribute to the biodistribution difficulties encountered by most cancer therapeutic agents. Antisense oligodeoxynucleotides (ASO) are a class of therapeutics where limited information is available about their distribution within a solid tumor environment. EXPERIMENTAL DESIGN: To assess ASO distribution, a fluorescein-labeled phosphorothionated ASO based on the G3139 mismatch control was injected systemically (i.v.) into tumor-bearing severe combined immunodeficient mice. Hoechst 33342 was injected i.v. to visualize active vasculature. Unstained sections were imaged through tiled fluorescence stereomicroscopy and then quantitated using novel algorithms. Tumor sections from four human tumor models were examined (CaSki, DU-145, C666-1, and C15) for hypoxia, apoptosis/necrosis, and morphology. RESULTS: For all four tumors, ASO accumulated within regions of hypoxia, necrosis, and apoptosis. Scatter plots of ASO versus active vasculature generated for each individual tumor revealed a consistent pattern of distribution of the ASO within each model. In C666-1 xenografts, the slopes of these scatter plots were significantly reduced from 0.41 to 0.16 when pretreated with the antivascular agent ZD6126 48 h before ASO injection. This was accompanied by the formation of large disseminated necrotic regions in the tumor, along with a 13.1 mmHg reduction in interstitial fluid pressure. CONCLUSIONS: These data suggest the possibility that these algorithms might offer a generalizable and objective methodology to describe the distribution of molecular therapeutic agents within a tumor microenvironment and to quantitatively assess distribution changes in response to combination therapies. PMID- 17908991 TI - Bortezomib inhibits nuclear factor-kappaB dependent survival and has potent in vivo activity in mesothelioma. AB - PURPOSE: Purpose of this study has been the assessment of nuclear factor-kappaB (NF-kappaB) as a survival factor in human mesothelial cells (HMC), transformed HMC and malignant mesothelioma (MMe) cells. We aimed at verifying whether the proteasome inhibitor Bortezomib could abrogate NF-kappaB activity in MMe cells, leading to tumor cell death and may be established as a novel treatment for this aggressive neoplasm. EXPERIMENTAL DESIGN: In HMC and MMe cells, NF-kappaB nuclear translocation and DNA binding were studied by electrophoretic mobility shift assay, following treatment with tumor necrosis factor-alpha (TNF-alpha). The IKK inhibitor Bay11-7082 was also tested to evaluate its effects on HMC, transformed HMC, and MMe cell viability upon exposure to asbestos fibers. Following Bortezomib treatment, cytotoxicity of MMe cells was evaluated by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, whereas apoptosis and cell cycle blockade were investigated by high-content analysis. Bortezomib was also given to mice bearing i.p. xenografts of MMe cells, and its effects on tumor growth were evaluated. RESULTS: Here, we show that NF-kappaB activity is a constitutive survival factor in transformed HMC, MMe cells, and acts as a survival factor in HMC exposed to asbestos fibers. Bortezomib inhibits NF-kappaB activity in MMe cells and induces cell cycle blockade and apoptosis in vitro as well as tumor growth inhibition in vivo. CONCLUSIONS: Inhibition of NF-kappaB constitutive activation in MMe cells by Bortezomib resulted in in vitro cytotoxicity along with apoptosis and in vivo tumor regression. Our results support the use of Bortezomib in the treatment of MMe and has led to a phase II clinical trial currently enrolling in Europe. PMID- 17908992 TI - N-acetylcysteine protects melanocytes against oxidative stress/damage and delays onset of ultraviolet-induced melanoma in mice. AB - PURPOSE: UV radiation is the major environmental risk factor for melanoma and a potent inducer of oxidative stress, which is implicated in the pathogenesis of several malignancies. We evaluated whether the thiol antioxidant N-acetylcysteine (NAC) could protect melanocytes from UV-induced oxidative stress/damage in vitro and from UV-induced melanoma in vivo. EXPERIMENTAL DESIGN: In vitro experiments used the mouse melanocyte line melan-a. For in vivo experiments, mice transgenic for hepatocyte growth factor and survivin, shown previously to develop melanoma following a single neonatal dose of UV irradiation, were given NAC (7 mg/mL; mother's drinking water) transplacentally and through nursing until 2 weeks after birth. RESULTS: NAC (1-10 mmol/L) protected melan-a cells from several UV-induced oxidative sequelae, including production of intracellular peroxide, formation of the signature oxidative DNA lesion 8-oxoguanine, and depletion of free reduced thiols (primarily glutathione). Delivery of NAC reduced thiol depletion and blocked formation of 8-oxoguanine in mouse skin following neonatal UV treatment. Mean onset of UV-induced melanocytic tumors was significantly delayed in NAC treated compared with control mice (21 versus 14 weeks; P = 0.0003). CONCLUSIONS: Our data highlight the potential importance of oxidative stress in the pathogenesis of melanoma and suggest that NAC may be useful as a chemopreventive agent. PMID- 17908993 TI - Association of RNASEL variants with prostate cancer risk in Hispanic Caucasians and African Americans. AB - PURPOSE: The RNASEL gene at 1q25 has been identified as a hereditary prostate cancer susceptibility gene, but to date, no study has investigated the role of RNASEL variants in Hispanic Caucasian men with prostate cancer. EXPERIMENTAL DESIGN: Two RNASEL common variants, located at amino acids 462 and 541, were genotyped in non-Hispanic Caucasian, Hispanic Caucasian, and African American prostate cancer cases and controls. RESULTS: The RNASEL 462 AA genotype was found to increase prostate cancer risk over 4-fold in Hispanic Caucasians [odds ratio (OR), 4.43; 95% confidence interval (95% CI), 1.68-11.68; P = 0.003] and over 10 fold in African Americans (OR, 10.41; 95% CI, 2.62-41.40; P = 0.001) when compared with the GG genotype. Analysis of the RNASEL 541 variant showed that Hispanic Caucasian patients with the GG genotype had a statistically significant increase in their risk for developing prostate cancer when compared with the TT and GT genotypes (OR, 1.91; 95% CI, 1.16-3.14; P = 0.01). A common G-T haplotype for the combination of the RNASEL 462 and 541 variants was found to occur more frequently in controls compared with cases in African Americans (P = 0.04) but not in non-Hispanic Caucasians or Hispanic Caucasians. CONCLUSIONS: This is the first study that investigates the association of prostate cancer risk with RNASEL variants in Hispanic men. Our data support the role of RNASEL as a predisposition gene for prostate cancer and showed a significant association between the RNASEL 462 variant and prostate cancer risk in African Americans and Hispanic Caucasians. PMID- 17908994 TI - Exisulind in combination with celecoxib modulates epidermal growth factor receptor, cyclooxygenase-2, and cyclin D1 against prostate carcinogenesis: in vivo evidence. AB - PURPOSE: Nonsteroidal anti-inflammatory drugs mediate anticancer effects by modulating cyclooxygenase-2 (COX-2)-dependent and/or COX-2-independent mechanism(s); however, the toxicity issue is a concern with single agents at higher doses. In this study, we determined the combined effect of celecoxib, a COX-2 inhibitor, along with exisulind (sulindac sulfone/Aptosyn) at low doses in prostate cancer. EXPERIMENTAL DESIGN: We used a sequential regimen of N-methyl-N nitrosourea + testosterone to induce prostate cancer in Wistar-Unilever rats. Following carcinogen treatment, celecoxib and exisulind individually and their combination at low doses were given in NIH-07 diet for 52 weeks. We determined the incidence of prostatic intraepithelial neoplasia, adenocarcinomas, rate of tumor cell proliferation, and apoptosis. Immunohistochemical and Western blot analysis were done to determine COX-2, epidermal growth factor receptor (EGFR), Akt, androgen receptor, and cyclin D1 expression. Serum prostaglandin E2 and tumor necrosis factor-alpha levels were determined using enzyme immunoassay/ELISA assays. RESULTS: The rats that received celecoxib in combination with exisulind at low doses showed a significant decrease in prostatic intraepithelial neoplasia and adenocarcinomas as well as an enhanced rate of apoptosis. An overall decrease in COX-2, EGFR, Akt, androgen receptor, and cyclin D1 expression was found associated with tumor growth inhibition. Reduced serum levels of COX-2 protein, prostaglandin E2, and tumor necrosis factor-alpha indicated anti-inflammatory effects. A strong inhibition of total and phosphorylated form of EGFR (Tyr(992) and Tyr(845)) and Akt (Ser(473)) was significant in rats given with these agents in combination. CONCLUSIONS: In this study, we show for the first time that the combination of celecoxib with exisulind at low doses could prevent prostate carcinogenesis by altering key molecular events. PMID- 17908995 TI - Genetic variants in cell cycle control pathway confer susceptibility to lung cancer. AB - PURPOSE: To test the hypothesis that common sequence variants of cell cycle control genes may affect lung cancer predisposition. EXPERIMENTAL DESIGN: We explored lung cancer risk associations of 11 polymorphisms in seven cell cycle genes in a large case-control study including 1,518 Caucasian lung cancer patients and 1,518 controls. RESULTS: When individuals with variant-containing genotypes were compared with homozygous wild-type carriers, a significantly increased lung cancer risk was identified for polymorphisms in p53 intron 6 [rs1625895; odds ratio (OR), 1.29; 95% confidence interval (95% CI), 1.08-1.55] and in p27 5' untranslated region (UTR; rs34330; OR, 1.27; 95% CI, 1.01-1.60). Compared with homozygous wild-types, the homozygous variant genotypes of STK15 F31I and CCND1 G870A were associated with a significantly altered lung cancer risk with ORs of 0.58 (95% CI, 0.37-0.90) and 1.26 (95% CI, 1.03-1.53), respectively. To assess the cumulative effects of all the investigated polymorphisms on lung carcinogenesis, we conducted a combined analysis and found that compared with low-risk individuals with few adverse alleles, individuals with more adverse alleles had an increased risk in a significant dose-dependent manner (P(trend) = 0.041). This pattern was more evident in ever smokers (P(trend) = 0.037), heavy smokers (P(trend) = 0.020), and older subjects (P(trend) = 0.011). Higher-order gene-gene interactions were evaluated using the classification and regression tree analysis, which indicated that STK15 F31I and p53 intron 6 polymorphisms might be associated with lung carcinogenesis in never/light-smokers and heavy smokers, respectively. CONCLUSIONS: Our results suggest that cell cycle gene polymorphisms and smoking may function collectively to modulate the risk of lung cancer. PMID- 17908996 TI - A new central scaffold for metastasis: parsing HEF1/Cas-L/NEDD9. AB - Greater understanding of metastasis is required to improve cancer treatment outcomes. Recently, changes in expression of the scaffold protein HEF1/CAS L/NEDD9 were found to be a potent prometastatic stimulus in melanoma and other cancers. Mechanistic studies suggest diverse cellular roles of HEF1 and highlight its importance in the response to extracellular cues that drive invasion and metastasis. As a metastatic "hub" for signaling in cancer, HEF1 may provide a useful target for drug discovery efforts. PMID- 17908997 TI - Cancer stem cells in radiation resistance. AB - Highly tumorigenic subpopulations of several solid cancers share characteristics with somatic stem cells. We showed recently that cancer stem cells, or tumor initiating cells, derived from human glioblastoma surgical specimens and xenografts display resistance to radiation due to increased activation of the DNA damage checkpoint. We additionally showed that these same tumor subpopulations promote tumor angiogenesis through increased expression of vascular endothelial growth factor. These studies and subsequent reports from other researchers support critical roles for cancer stem cells in determining tumor response to therapy. Hypoxia and stem cell maintenance pathways may provide therapeutic targets to sensitize cancer stem cells to cytotoxic therapies to improve cancer patient treatments. PMID- 17908998 TI - The fuzzy math of solid tumor stem cells: a perspective. AB - Apparently effective therapeutic agents very often fail to cure cancer patients. It is therefore attractive to wonder whether a specific resistant cell subset should be recognized and separately targeted. In solid tumors, such as carcinomas, a minor population of "cancer stem cells" has been proposed and sought experimentally in human tumors and isolated cell populations. It is often overlooked that the rationale and supportive data are essentially numerical and can be evaluated as such. A reevaluation of the published studies and related claims within awarded U.S. patents suggests that the mathematical support for the concept of therapeutically useful stem cells is weak and may even invalidate the foundations of these publications and patent claims. Mathematical arguments should be used more consistently, because they can serve as a guide for interpreting studies into cancer stem cells of solid tumors. PMID- 17908999 TI - MicroRNA-21 knockdown disrupts glioma growth in vivo and displays synergistic cytotoxicity with neural precursor cell delivered S-TRAIL in human gliomas. AB - Despite the development of new glioma therapies that allow for tumor-targeted in situ delivery of cytotoxic drugs, tumor resistance to apoptosis remains a key impediment to effective treatment. Mounting evidence indicates that microRNAs (miRNA) might play a fundamental role in tumorigenesis, controlling cell proliferation and apoptosis. In gliomas, microRNA-21 (miR-21) levels have been reported to be elevated and their knockdown is associated with increased apoptotic activity. We hypothesized that suppression of miR-21 might sensitize gliomas for cytotoxic tumor therapy. With the use of locked nucleic acid (LNA) antimiR-21 oligonucleotides, bimodal imaging vectors, and neural precursor cells (NPC) expressing a secretable variant of the cytotoxic agent tumor necrosis factor-related apoptosis inducing ligand (S-TRAIL), we show that the combined suppression of miR-21 and NPC-S-TRAIL leads to a synergistic increase in caspase activity and significantly decreased cell viability in human glioma cells in vitro. This phenomenon persists in vivo, as we observed complete eradication of LNA-antimiR-21-treated gliomas subjected to the presence of NPC-S-TRAIL in the murine brain. Our results reveal the efficacy of miR-21 antagonism in murine glioma models and implicate miR-21 as a target for therapeutic intervention. Furthermore, our findings provide the basis for developing combination therapies using miRNA modulation and cytotoxic tumor therapies. PMID- 17909000 TI - Evidence for calpain-mediated androgen receptor cleavage as a mechanism for androgen independence. AB - Prostate carcinoma is the most commonly diagnosed cancer in men and the second leading cause of death due to cancer in Western civilization. Androgen ablation therapy is effective in treating androgen-dependent tumors, but eventually, androgen-independent tumors recur and are refractory to conventional chemotherapeutics. Hence, the emergence of androgen independence is the most challenging problem in managing prostate tumors. We report a novel mechanism of androgen independence: calpain cleaves the androgen receptor (AR) into an androgen-independent isoform. In vitro and in vivo analyses show that calpain removes the COOH-terminal ligand binding domain generating a constitutively active molecule. Analysis of human prostate tumors indicates that several tumors express higher levels of this truncated AR than noncancerous prostate tissue. In transient transfection studies, the truncated AR is three to five times more potent than the full-length receptor in transactivating transcription. The androgen-independent Rv1 cells express high levels of the truncated AR, and treatment of these cells with a calpain inhibitor reduces truncated AR expression. In the absence of androgen, inhibition of calpain activity induces apoptosis. The HIV protease inhibitor amprenavir inhibits calpain activity and is also effective in inducing apoptosis in the Rv1 cell line. The cell culture studies were reproduced in a mouse xenograft model, where, in the absence of androgens, amprenavir significantly reduces tumor growth. Together, these studies indicate that calpain-dependent proteolysis of the AR may be a mechanism of androgen independence. The calpain inhibition studies suggest that inhibiting this activity may be a potential treatment for some androgen-independent prostate tumors. PMID- 17909002 TI - Loss of the mitochondrial bioenergetic capacity underlies the glucose avidity of carcinomas. AB - The down-regulation of the catalytic subunit of the mitochondrial H+-ATP synthase (beta-F1-ATPase) is a hallmark of most human carcinomas. This characteristic of the cancer cell provides a proteomic signature of cellular bioenergetics that can predict the prognosis of colon, lung, and breast cancer patients. Here we show that the in vivo tumor glucose uptake of lung carcinomas, as assessed by positron emission tomography in 110 patients using 2-deoxy-2-[18F]fluoro-d-glucose as probe, inversely correlates with the bioenergetic signature determined by immunohistochemical analysis in tumor surgical specimens. Further, we show that inhibition of the activity of oxidative phosphorylation by incubation of cancer cells with oligomycin triggers a rapid increase in their rates of aerobic glycolysis. Moreover, we show that the cellular expression level of the beta-F1 ATPase protein of mitochondrial oxidative phosphorylation inversely correlates (P < 0.001) with the rates of aerobic glycolysis in cancer cells. The results highlight the relevance of the alteration of the bioenergetic function of mitochondria for glucose capture and consumption by aerobic glycolysis in carcinomas. PMID- 17909001 TI - FBXW7/hCDC4 is a general tumor suppressor in human cancer. AB - The ubiquitin-proteasome system is a major regulatory pathway of protein degradation and plays an important role in cellular division. Fbxw7 (or hCdc4), a member of the F-box family of proteins, which are substrate recognition components of the multisubunit ubiquitin ligase SCF (Skp1-Cdc53/Cullin-F-box protein), has been shown to mediate the ubiquitin-dependent proteolysis of several oncoproteins including cyclin E1, c-Myc, c-Jun, and Notch. The oncogenic potential of Fbxw7 substrates, frequent allelic loss in human cancers, and demonstration that mutation of FBXW7 cooperates with p53 in mouse tumorigenesis have suggested that Fbxw7 could function as a tumor suppressor in human cancer. Here, we carry out an extensive genetic screen of primary tumors to evaluate the role of FBXW7 as a tumor suppressor in human tumorigenesis. Our results indicate that FBXW7 is inactivated by mutation in diverse human cancer types with an overall mutation frequency of approximately 6%. The highest mutation frequencies were found in tumors of the bile duct (cholangiocarcinomas, 35%), blood (T-cell acute lymphocytic leukemia, 31%), endometrium (9%), colon (9%), and stomach (6%). Approximately 43% of all mutations occur at two mutational "hotspots," which alter Arg residues (Arg465 and Arg479) that are critical for substrate recognition. Furthermore, we show that Fbxw7Arg465 hotspot mutant can abrogate wild-type Fbxw7 function through a dominant negative mechanism. Our study is the first comprehensive screen of FBXW7 mutations in various human malignancies and shows that FBXW7 is a general tumor suppressor in human cancer. PMID- 17909003 TI - Trastuzumab-resistant HER2-driven breast cancer cells are sensitive to epigallocatechin-3 gallate. AB - Overexpression of the epidermal growth factor receptor family member HER2 is found in approximately 30% of breast cancers and is a target for immunotherapy. Trastuzumab, a humanized monoclonal antibody against HER2, is cytostatic when added alone and highly successful in clinical settings when used in combination with other chemotherapeutic agents. Unfortunately, HER2 tumors in patients develop resistance to trastuzumab or metastasize to the brain, which is inaccessible to antibody therapy. Previously, we showed that the green tea polyphenol epigallocatechin-3 gallate (EGCG) inhibits growth and transformed phenotype of Her-2/neu-driven mouse mammary tumor cells. The different modes of action of EGCG and trastuzumab led us to hypothesize that EGCG will inhibit HER2 driven breast cancer cells resistant to trastuzumab. We studied trastuzumab resistant BT474 human breast cancer cells, isolated by chronic trastuzumab exposure, and JIMT-1 breast cancer cells, derived from a pleural effusion in a patient who displayed clinical resistance to trastuzumab therapy. EGCG treatment caused a dose-dependent decrease in growth and cellular ATP production, and apoptosis at high concentrations. Akt activity was suppressed by EGCG leading to the induction of FOXO3a and target cyclin-dependent kinase inhibitor p27Kip1 levels. Thus, EGCG in combination with trastuzumab may provide a novel strategy for treatment of HER2-overexpressing breast cancers, given that EGCG can cross the blood-brain barrier. PMID- 17909005 TI - Opposing function of the proprotein convertases furin and PACE4 on breast cancer cells' malignant phenotypes: role of tissue inhibitors of metalloproteinase-1. AB - Proteolytic cleavage of various cancer-related substrates by the proprotein convertases (PC) was reported to be important in the processes of neoplasia. These enzymes are inhibited by their naturally occurring inhibitors, the prosegments (ppPC), and by the engineered general PC inhibitor, the serpin variant alpha1-PDX. In the present study, we sought to compare the effect of these PC inhibitors on malignant phenotypes of breast cancer cells. Overexpression in a stable manner of alpha1-PDX and the prosegment ppPACE4 in MDA MB-231 breast cancer cells resulted in increased matrix metalloproteinase (MMP)-9 (but not MMP-2) activity and a reduced secretion of tissue inhibitor of metalloproteinase 1 (TIMP-1). This was associated with significant enhancement in cell motility, migration, and invasion of collagen in vitro. In contrast, ppFurin expression in these cells decreased MMP-9 activity and diminished these biological functions, but had no significant effect on TIMP-1 secretion. Taken together, these data showed the specific and opposing roles of Furin and PACE4 in the regulation of MMP-9/TIMP-1-mediated cell motility and invasion. PMID- 17909004 TI - Glucuronidation of nicotine and cotinine by UGT2B10: loss of function by the UGT2B10 Codon 67 (Asp>Tyr) polymorphism. AB - Nicotine, the major addicting agent in tobacco and tobacco smoke, undergoes a complex metabolic pathway, with approximately 22% of nicotine urinary metabolites in the form of phase II N-glucuronidated compounds. Recent studies have shown that UGT2B10 is a major enzyme involved in the N-glucuronidation of several tobacco-specific nitrosamines. In the present study, microsomes of UGT2B10 overexpressing HEK293 cells exhibited high N-glucuronidation activity against both nicotine and cotinine with apparent KM's that were 37- and 3-fold lower than that observed for microsomes of UGT1A4-overexpressing cells against nicotine and cotinine, respectively. The KM of microsomes from wild-type (WT) UGT2B10 overexpressing cells for nicotine and cotinine was similar to that observed for human liver microsomes (HLM) against both substrates. The level of glucuronidated nicotine or cotinine in 112 HLM samples was correlated with UGT2B10 genotype; the levels of nicotine- and cotinine-glucuronide were 21% to 30% lower in specimens from subjects with the UGT2B10 (*1/*2) genotype compared with specimens from subjects with the WT UGT2B10 (*1/*1) genotype; a 5- and 16-fold lower level of nicotine- and cotinine-glucuronide formation, respectively, was observed in HLM from subjects with the UGT2B10 (*2/*2) genotype. In contrast to the relatively high activity observed for cells overexpressing WT UGT2B10 in vitro, little or no glucuronidation was observed for microsomes from cells overexpressing the UGT2B10*2 variant against either nicotine or cotinine. These data suggest that UGT2B10 is the major hepatic enzyme involved in nicotine/cotinine glucuronidation and that the UGT2B10*2 variant significantly reduces nicotine- and cotinine-N glucuronidation formation and plays an important role in nicotine metabolism and elimination. PMID- 17909006 TI - Fetal microchimerism in women with breast cancer. AB - Fetal microchimerism (FMc) describes long-term persistence of small numbers of fetal-derived allogeneic cells in the mother. Although FMc has been implicated as a mechanism of autoimmune disease, it may confer a beneficial effect with immune surveillance of malignant cells. We hypothesized that allogeneic FMc imparts a protective effect against breast cancer. Two observations provided a rationale for the study hypothesis. First, allogeneic cells convey risk reduction for recurrent malignancy in hematopoietic cell transplantation. Second, reduced risk of breast cancer is well recognized among parous compared with nulliparous women. As an initial test of the hypothesis, we investigated 82 women, 35 with breast cancer and 47 who were healthy, for male DNA in peripheral blood, presumed from a prior pregnancy with a male fetus. The prevalence and levels of male DNA were determined by real-time quantitative PCR for the Y chromosome-specific gene DYS14 in DNA extracted from peripheral blood mononuclear cells. FMc was found significantly more often in healthy women than women with breast cancer (43% versus 14%, respectively). Considering the absence of FMc as a risk factor, the odds ratio was 4.4 [95% confidence intervals (95% CI), 1.34-16.99; P = 0.006]. Restricting analysis to women known to had given birth to a son, the odds ratio was 5.9 (95% CI, 1.26-6.69; P = 0.01). Our findings indicate that allogeneic FMc may contribute to reduction in risk of breast cancer. Further studies are indicated and, if confirmed, extended studies to examine whether allogeneic immune surveillance from FMc is deficient in women with breast cancer. PMID- 17909007 TI - A novel variant of ileal bile acid binding protein is up-regulated through nuclear factor-kappaB activation in colorectal adenocarcinoma. AB - Ileal bile acid binding protein (IBABP) is the only cytosolic protein known to bind and transport bile acids. Because IBABP is reportedly up-regulated in colorectal cancer, it has been suggested as a link between bile acids and the risk of colorectal cancer. However, in this study, we show that IBABP is not up regulated. Rather, a novel transcript of the IBABP gene, which encodes an additional 49 NH(2)-terminal amino acid residues, is up-regulated in colorectal cancer (P < 0.001). The novel transcript, called IBABP-L, is also distinct from IBABP because its transcription is controlled by nuclear factor-kappaB (NF kappaB) rather than by the farnesoid X receptor. Most significantly, IBABP-L is necessary for the survival of HCT116 colon cancer cells in the presence of physiologic levels of the secondary bile acid deoxycholate. Collectively, the studies point toward a unique bile acid response pathway involving NF-kappaB and IBABP-L that could be useful for diagnosis and could potentially be targeted for therapeutic benefit. PMID- 17909008 TI - Reduced body size and decreased intestinal tumor rates in HDAC2-mutant mice. AB - Histone deacetylases (HDAC) reverse the acetylation of histone and nonhistone proteins and thereby modulate chromatin structure and function of nonhistone proteins. Many tumor cell lines and experimental tumors respond to HDAC inhibition. To assess the role of an individual HDAC isoenzyme in physiology and tumor development, HDAC2-mutant mice were generated from a gene trap embryonic stem cell clone. These mice express a catalytically inactive fusion protein of the NH(2)-terminal part of HDAC2 and beta-galactosidase, which fails to integrate into corepressor complexes with mSin3B. They are the first class 1 HDAC mutant mice that are viable although they are approximately 25% smaller than their littermates. Cell number and thickness of intestinal mucosa are reduced. Mutant embryonic fibroblasts fail to respond to insulin-like growth factor I (IGF) by the IGF-I-induced increase in cell number observed in wild-type cells. These data suggest a novel link between HDACs and IGF-I-dependent responses. Crossing of HDAC2-mutant with tumor-prone APC(min) mice revealed tumor rates that are lower in HDAC2-deficient mice by 10% to 100% depending on segment of the gut and sex of the mice. These mice provide evidence that the key functions of HDAC2, although not essential for survival of the organism, play a rate-limiting role for tumor development in vivo. PMID- 17909009 TI - High cancer-specific expression of mesothelin (MSLN) is attributable to an upstream enhancer containing a transcription enhancer factor dependent MCAT motif. AB - Identification of genes with cancer-specific overexpression offers the potential to efficiently discover cancer-specific activities in an unbiased manner. We apply this paradigm to study mesothelin (MSLN) overexpression, a nearly ubiquitous, diagnostically and therapeutically useful characteristic of pancreatic cancer. We identified an 18-bp upstream enhancer, termed CanScript, strongly activating transcription from an otherwise weak tissue-nonspecific promoter and operating selectively in cells having aberrantly elevated cancer specific MSLN transcription. Introducing mutations into CanScript showed two functionally distinct sites: an Sp1-like site and an MCAT element. Gel retardation and chromatin immunoprecipitation assays showed the MCAT element to be bound by transcription enhancer factor (TEF)-1 (TEAD1) in vitro and in vivo. The presence of TEF-1 was required for MSLN protein overexpression as determined by TEF-1 knockdown experiments. The cancer specificity seemed to be provided by a putative limiting cofactor of TEF-1 that could be outcompeted by exogenous TEF-1 only in a MSLN-overexpressing cell line. A CanScript concatemer offered enhanced activity. These results identify a TEF family member as a major regulator of MSLN overexpression, a fundamental characteristic of pancreatic and other cancers, perhaps due to an upstream and highly frequent aberrant cellular activity. The CanScript sequence represents a modular element for cancer-specific targeting, potentially suitable for nearly a third of human malignancies. PMID- 17909011 TI - Cisplatin depletes TREX2 and causes Robertsonian translocations as seen in TREX2 knockout cells. AB - Cisplatin, an anticancer drug, forms DNA interstrand cross-links (ICL) that interfere with replication, whereas TREX2 is a 3'-->5' exonuclease that removes 3' mismatched nucleotides and promotes cellular proliferation. Here, we show that TREX2 is depleted in human cells derived from cancer after exposure to cisplatin but not other genotoxins including another cross-linking agent, mitomycin C (MMC), indicating a potential role for TREX2 depletion in cisplatin-induced cytotoxicity. To better understand TREX2 cellular function, we deleted TREX2 in mouse embryonic stem (ES) cells by gene targeting and find these cells exhibit reduced proliferation and gross chromosomal rearrangements including Robertsonian translocations (RbT). Quite interestingly, ES cells exposed to cisplatin also exhibit RbTs. By contrast, RbTs are not observed for ES cells exposed to MMC, indicating that RbTs are not caused by ICLs but instead TREX2 depletion by either cisplatin exposure or mutation. Taken together, our results show that cisplatin depletes TREX2 and causes genomic instability that is similarly observed in TREX2 mutant cells. Thus, cisplatin has two potential cytotoxic activities: (a) the generation of ICLs and (b) the depletion of TREX2. PMID- 17909010 TI - Epidermal growth factor receptor cooperates with signal transducer and activator of transcription 3 to induce epithelial-mesenchymal transition in cancer cells via up-regulation of TWIST gene expression. AB - Aberrant epidermal growth factor receptor (EGFR) signaling is a major cause of tumor progression and metastasis; the underlying mechanisms, however, are not well understood. In particular, it remains elusive whether deregulated EGFR pathway is involved in epithelial-mesenchymal transition (EMT), an early event that occurs during metastasis of cancers of an epithelial origin. Here, we show that EGF induces EGFR-expressing cancer cells to undergo a transition from the epithelial to the spindle-like mesenchymal morphology. EGF reduced E-cadherin expression and increased that of mesenchymal proteins. In search of a downstream mediator that may account for EGF-induced EMT, we focused on transcription repressors of E-cadherin, TWIST, SLUG, and Snail and found that cancer cells express high levels of TWIST and that EGF enhances its expression. EGF significantly increases TWIST transcripts and protein in EGFR-expressing lines. Forced expression of EGFR reactivates TWIST expression in EGFR-null cells. TWIST expression is suppressed by EGFR and Janus-activated kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) inhibitors, but not significantly by those targeting phosphoinositide-3 kinase and MEK/ERK. Furthermore, constitutively active STAT3 significantly activates the TWIST promoter, whereas the JAK/STAT3 inhibitor and dominant-negative STAT3 suppressed TWIST promoter. Deletion/mutation studies further show that a 26-bp promoter region contains putative STAT3 elements required for the EGF-responsiveness of the TWIST promoter. Chromatin immunoprecipitation assays further show that EGF induces binding of nuclear STAT3 to the TWIST promoter. Immunohistochemical analysis of 130 primary breast carcinomas indicates positive correlations between non-nuclear EGFR and TWIST and between phosphorylated STAT3 and TWIST. Together, we report here that EGF/EGFR signaling pathways induce cancer cell EMT via STAT3 mediated TWIST gene expression. PMID- 17909012 TI - Pediatric KIT wild-type and platelet-derived growth factor receptor alpha-wild type gastrointestinal stromal tumors share KIT activation but not mechanisms of genetic progression with adult gastrointestinal stromal tumors. AB - Fewer than 15% of gastrointestinal stromal tumors (GIST) in pediatric patients harbor KIT or platelet-derived growth factor receptor alpha (PDGFRA) mutations in contrast to a mutation rate of 80% in adult GISTs. However, some therapeutic inhibitors of KIT have efficacy in pediatric GIST, suggesting that KIT may, nevertheless, play an important role in oncogenesis. In adult GIST, characteristic cytogenetic changes occur during progression to malignancy. A better understanding of mechanisms of genetic progression and KIT and PDGFRA transforming roles in pediatric GIST might facilitate treatment advances. KIT and PDGFRA mutation analysis was done in 27 pediatric GISTs. The activation status of KIT, PDGFRA, and downstream signaling intermediates was defined, and chromosomal aberrations were determined by single nucleotide polymorphism assays. Mutations in KIT or PDGFRA were identified in 11% of pediatric GISTs. KIT and the signaling intermediates AKT and mitogen-activated protein kinase were activated in pediatric GISTs. In particular, most pediatric KIT-wild-type GISTs displayed levels of KIT activation similar to levels in adult KIT-mutant GISTs. Pediatric KIT-wild-type GISTs lacked the typical cytogenetic deletions seen in adult KIT mutant GISTs. Notably, most pediatric KIT-wild-type GISTs progress to malignancy without acquiring large-scale chromosomal aberrations, which is a phenomenon not reported previously in malignant solid tumors. KIT activation levels in pediatric KIT-wild-type GISTs are comparable with those in KIT-mutant GISTs. Therapies that inhibit KIT activation, or crucial KIT signaling intermediates, should be explored in pediatric KIT-wild-type GIST. PMID- 17909013 TI - Prolonged exposure to reduced levels of androgen accelerates prostate cancer progression in Nkx3.1; Pten mutant mice. AB - In this report, we have investigated the relationship between androgen levels and prostate tumorigenesis in Nkx3.1; Pten mutant mice, a genetically engineered mouse model of human prostate cancer. By experimentally manipulating serum levels of testosterone in these mice for an extended period (i.e., 7 months), we have found that prolonged exposure of Nkx3.1; Pten mutant mice to androgen levels that are 10-fold lower than normal (the "Low-T" group) resulted in a marked acceleration of prostate tumorigenesis compared with those exposed to androgen levels within the reference range (the "Normal-T" group). We found that prostate tumors from the Low-T mutant mice share a similar gene expression profile as androgen-independent prostate tumors from these mutant mice, which includes the deregulated expression of several genes that are up-regulated in human hormone refractory prostate cancer, such as Vav3 and Runx1. We propose that exposure to reduced androgens may promote prostate tumorigenesis by selecting for molecular events that promote more aggressive, hormone-refractory tumors. PMID- 17909014 TI - The Ajuba LIM domain protein is a corepressor for SNAG domain mediated repression and participates in nucleocytoplasmic Shuttling. AB - The SNAG repression domain is comprised of a highly conserved 21-amino acid sequence, is named for its presence in the Snail/growth factor independence-1 class of zinc finger transcription factors, and is present in a variety of proto oncogenic transcription factors and developmental regulators. The prototype SNAG domain containing oncogene, growth factor independence-1, is responsible for the development of T cell thymomas. The SNAIL proteins also encode the SNAG domain and play key roles in epithelial mesenchymal differentiation events during development and metastasis. Significantly, these oncogenic functions require a functional SNAG domain. The molecular mechanisms of SNAG domain-mediated transcriptional repression are largely unknown. Using a yeast two-hybrid strategy, we identified Ajuba, a multiple LIM domain protein that can function as a corepressor for the SNAG domain. Ajuba interacts with the SNAG domain in vitro and in vivo, colocalizes with it, and enhances SNAG-mediated transcriptional repression. Ajuba shuttles between the cytoplasm and the nucleus and may form a novel intracellular signaling system. Using an integrated reporter gene combined with chromatin immunoprecipitation, we observed rapid, SNAG-dependent assembly of a multiprotein complex that included Ajuba, SNAG, and histone modifications consistent with the repressed state. Thus, SNAG domain proteins may bind Ajuba, trapping it in the nucleus where it functions as an adapter or molecular scaffold for the assembly of macromolecular repression complexes at target promoters. PMID- 17909015 TI - Epigenetic inactivation of a cluster of genes flanking MLH1 in microsatellite unstable colorectal cancer. AB - Biallelic promoter methylation and transcriptional silencing of the MLH1 gene occurs in the majority of sporadic colorectal cancers exhibiting microsatellite instability due to defective DNA mismatch repair. Long-range epigenetic silencing of contiguous genes has been found on chromosome 2q14 in colorectal cancer. We hypothesized that epigenetic silencing of MLH1 could occur on a regional scale affecting additional genes within 3p22, rather than as a focal event. We studied the levels of CpG island methylation and expression of multiple contiguous genes across a 4 Mb segment of 3p22 including MLH1 in microsatellite-unstable and stable cancers, and their paired normal colonic mucosa. We found concordant CpG island hypermethylation, H3-K9 dimethylation and transcriptional silencing of MLH1 and multiple flanking genes spanning up to 2.4 Mb in microsatellite-unstable colorectal cancers. This region was interspersed with unmethylated genes, which were also transcriptionally repressed. Expression of both methylated and unmethylated genes was reactivated by methyltransferase and histone deacetylase inhibitors in a microsatellite-unstable colorectal carcinoma cell line. Two genes at the telomeric end of the region were also hypermethylated in microsatellite stable cancers, adenomas, and at low levels in normal colonic mucosa from older individuals. Thus, the cluster of genes flanking MLH1 that was specifically methylated in the microsatellite-unstable group of cancers extended across 1.1 Mb. Our results show that coordinate epigenetic silencing extends across a large chromosomal region encompassing MLH1 in microsatellite-unstable colorectal cancers. Simultaneous epigenetic silencing of this cluster of 3p22 genes may contribute to the development or progression of this type of cancer. PMID- 17909016 TI - Targeted repression of bone morphogenetic protein 7, a novel target of the p53 family, triggers proliferative defect in p53-deficient breast cancer cells. AB - p53 tumor suppressor and its family members, p63 and p73, are known to play a role in the survival of cells exposed to stress signals. As a transcription factor, the p53 family proteins induce a plethora of target genes that mediate their functions in the cell cycle, apoptosis, and other biological activities. However, the mechanism by which the p53 family proteins regulate their cell survival functions is still not clear. Here, we showed that bone morphogenetic protein 7 (BMP7) is a novel target gene regulated by the p53 family and mediates the cell survival function of the basal physiologically relevant level of p53. Specifically, we found that knockdown of BMP7 markedly inhibits the proliferation of p53-deficient, but not p21-knockdown, breast cancer cells compared with the ones with wild-type p53. In addition, we found that inhibitor of differentiation or DNA binding 2 (Id2), a transcription factor implicated for cell survival, is regulated by the BMP7 and p53 pathways. Interestingly, whereas a functional BMP7 or p53 pathway is sufficient to maintain the basal level of Id2 expression, loss of both pathways abrogates Id2 expression. Furthermore, we showed that overexpression of Id2 can restore p53-deficient cells to survive in the absence of BMP7. As a result, we identified a previously unrecognized role for BMP7 in the maintenance of cell survival for p53-deficient cells, at least in part, through Id2. Together, we hypothesize that breast cancer patients with mutant p53 might benefit from targeted repression of BMP7 expression and/or targeted inhibition of the BMP7 pathway. PMID- 17909017 TI - Egr-1 mediates hypoxia-inducible transcription of the NDRG1 gene through an overlapping Egr-1/Sp1 binding site in the promoter. AB - N-myc down-regulated gene 1 (NDRG1/Cap43) is inducible by a variety of environmental stressors, including hypoxia. The present study identified a cis acting element mediating the transactivation of the NDRG1 gene in murine RAW264.7 macrophage cells treated with hypoxia or deferoxamine, an iron chelator mimicking hypoxia. Through a series of deletions of the promoter of NDRG1 luciferase constructs, a minimal cis-acting element conferring inducibility by hypoxia and deferoxamine was localized to an early growth response 1 (Egr-1) and Sp1 overlapping binding site. Electrophoretic mobility shift assay, antibody supershift assay, and mutations of the Egr-1 binding site confirmed the specific binding of Egr-1 protein to this Egr-1/Sp1 motif. In addition, hypoxia increased the level of Egr-1 protein that correlated with induction of NDRG1 expression at both RNA and protein levels. Transient transfection of the Egr-1 gene into HeLa cells also resulted in up-regulation of the NDRG1 mRNA. The role of Egr-1 was further verified by mutations in the Egr-1 binding site, which reduced promoter inducibility by hypoxia and deferoxamine. Furthermore, the induction of NDRG1 expression by hypoxia and deferoxamine was diminished by RNA interference knockdown of Egr-1 gene expression and in Egr-1-/- mouse embryonic fibroblasts (MEF) compared with Egr-1+/- MEFs. These results showed for the first time that Egr-1 regulates NDRG1 transcription through an overlapping Egr-1/Sp1 binding site that acts as a major site of positive regulation of the NDRG1 promoter by hypoxia signaling. PMID- 17909018 TI - A CDKN2A mutation in familial melanoma that abrogates binding of p16INK4a to CDK4 but not CDK6. AB - The CDKN2A locus encodes two distinct proteins, p16INK4a and p14ARF, both of which are implicated in replicative senescence and tumor suppression in different contexts. Here, we describe the characterization of a novel strain of human diploid fibroblasts (designated Milan HDFs) from an individual who is homozygous for the R24P mutation in p16INK4a. As this mutation occurs in the first exon of INK4a (exon 1alpha), it has no effect on the primary sequence of p14(ARF). Based on both in vitro and in vivo analyses, the R24P variant is specifically defective for binding to CDK4 but remains able to associate with CDK6. Nevertheless, Milan HDFs behave as if they are p16INK4a deficient, in terms of sensitivity to spontaneous and oncogene-induced senescence, and the R24P variant has little effect on proliferation when ectopically expressed in normal fibroblasts. It can, however, impair the proliferation of U20S cells, presumably because they express more CDK6 than primary fibroblasts. These observations suggest that CDK4 and CDK6 are not functionally redundant and underscore the importance of CDK4 in the development of melanoma. PMID- 17909019 TI - Human bone marrow derived mesenchymal stem cells do not undergo transformation after long-term in vitro culture and do not exhibit telomere maintenance mechanisms. AB - Significant improvement in the understanding of mesenchymal stem cell (MSC) biology has opened the way to their clinical use. However, concerns regarding the possibility that MSCs undergo malignant transformation have been raised. We investigated the susceptibility to transformation of human bone marrow (BM) derived MSCs at different in vitro culture time points. MSCs were isolated from BM of 10 healthy donors and propagated in vitro until reaching either senescence or passage (P) 25. MSCs in the senescence phase were closely monitored for 8 to 12 weeks before interrupting the cultures. The genetic characterization of MSCs was investigated through array-comparative genomic hybridization (array-CGH), conventional karyotyping, and subtelomeric fluorescent in situ hybridization analysis both before and after prolonged culture. MSCs were tested for the expression of telomerase activity, human telomerase reverse transcriptase (hTERT) transcripts, and alternative lengthening of telomere (ALT) mechanism at different passages. A huge variability in terms of proliferative capacity and MSCs life span was noted between donors. In eight of 10 donors, MSCs displayed a progressive decrease in proliferative capacity until reaching senescence. In the remaining two MSC samples, the cultures were interrupted at P25 to pursue data analysis. Array-CGH and cytogenetic analyses showed that MSCs expanded in vitro did not show chromosomal abnormalities. Telomerase activity and hTERT transcripts were not expressed in any of the examined cultures and telomeres shortened during the culture period. ALT was not evidenced in the MSCs tested. BM-derived MSCs can be safely expanded in vitro and are not susceptible to malignant transformation, thus rendering these cells suitable for cell therapy approaches. PMID- 17909020 TI - Cell division cycle 25B phosphatase is essential for benzo(a)pyrene-7,8-Diol-9,10 epoxide induced neoplastic transformation. AB - Cell division cycle 25B (Cdc25B) phosphatase controls entry into mitosis and regulates recovery from G2-M checkpoint-induced arrest. In the present study, we show that exposure of diploid mouse embryonic fibroblasts (MEF) to the ultimate carcinogen anti-benzo(a)pyrene (BP)-7,8-diol-9,10-epoxide (anti-BPDE) resulted in a concentration- and time-dependent increase in Cdc25B protein levels. Chronic exposure of wild-type (Cdc25B+/+) MEFs to anti-BPDE (0.1 micromol/L) caused neoplastic transformation characterized by colony formation in culture and tumor production in nude mice. In contrast, the Cdc25B null MEFs were resistant to anti BPDE-induced transformation. Furthermore, a carcinogenic dose of the parent hydrocarbon (BP) increased Cdc25B protein levels in the target organ, lung. The biological importance of elevated Cdc25B levels was documented by the early reentry into mitosis of cells overexpressing ectopic Cdc25B levels even in the presence of DNA damage following anti-BPDE exposure, whereas control cells resumed only after DNA damage was repaired. We conclude that Cdc25B has an essential role in anti-BPDE-induced neoplastic transformation, including regulation of cell cycle resumption in the presence of DNA damage. PMID- 17909021 TI - Reduction in IkappaB kinase alpha expression promotes the development of skin papillomas and carcinomas. AB - We reported recently a marked reduction in IkappaB kinase alpha (IKKalpha) expression in a large proportion of human poorly differentiated squamous cell carcinomas (SCC) and the occurrence of Ikkalpha mutations in human SCCs. In addition, overexpression of IKKalpha in the epidermis inhibited the development of skin carcinomas and metastases in mice. However, whether a reduction in IKKalpha expression promotes skin tumor development is currently unknown. Here, we assessed the susceptibility of Ikkalpha hemizygotes to chemical carcinogen induced skin carcinogenesis. Ikkalpha+/- mice developed 2 times more papillomas and 11 times more carcinomas than did Ikkalpha+/+ mice. The tumors were larger in Ikkalpha+/- than in Ikkalpha+/+ mice, but tumor latency was shorter in Ikkalpha+/ than in Ikkalpha+/+ mice. Some of the Ikkalpha+/- papillomas and most Ikkalpha+/ carcinomas lost the remaining Ikkalpha wild-type allele. Somatic Ikkalpha mutations were detected in carcinomas and papillomas. The chemical carcinogen induced H-Ras mutations were detected in all the tumors. The phorbol ester tumor promoter induced higher mitogenic and angiogenic activities in Ikkalpha+/- than in Ikkalpha+/+ skin. These elevated activities were intrinsic to keratinocytes, suggesting that a reduction in IKKalpha expression provided a selective growth advantage, which cooperated with H-Ras mutations to promote papilloma formation. Furthermore, excessive extracellular signal-regulated kinase and IKK kinase activities were observed in carcinomas compared with those in papillomas. Thus, the combined mitogenic, angiogenic, and IKK activities might contribute to malignant conversion. Our findings provide evidence that a reduction in IKKalpha expression promotes the development of papillomas and carcinomas and that the integrity of the Ikkalpha gene is required for suppressing skin carcinogenesis. PMID- 17909022 TI - Tenascin-W is a novel marker for activated tumor stroma in low-grade human breast cancer and influences cell behavior. AB - This is the first report about human tenascin-W, the fourth and final member of the extracellular matrix protein family of tenascins. Sixty-three human breast tumor extracts were analyzed by Western blotting for the presence of tenascin-W and compared with tenascin-C, an established marker of tumor stroma. Interestingly, we found tenascin-W expression in the majority of the tumor tissues, but no detectable expression in the normal mammary parenchyma. Eighty one percent of the breast tumor samples were tenascin-W positive and 86% showed expression of tenascin-C. However, tenascin-W and tenascin-C amounts varied greatly between tumors and some contained either tenascin-W or tenascin-C exclusively, indicating independent mechanisms regulating their expression. Although there was no difference between high- or low-grade tumors with respect to the presence of tenascin-C, tenascin-W was more prominent in low-grade tumors. For 42 of the breast cancer tissues, a frozen tumor microarray was available to confirm the Western blot data by immunohistochemistry. Similar to tenascin-C, tenascin-W was detected in the tumor stroma. Fibroblasts adhered to tenascin-W in a beta(1) integrin-dependent manner and spread with a distinctive morphology under conditions where they remained round on tenascin-C. CHOB2 cells expressing alpha(v)beta(1) or alpha4beta(1) integrins were able to spread on tenascin-W. Furthermore, addition of tenascin-W to the culture medium increased migration of breast cancer cells toward a fibronectin substratum in vitro. These data imply that tenascin-W expression in the activated tumor stroma facilitates tumorigenesis by supporting the migratory behavior of breast cancer cells. PMID- 17909023 TI - In vivo imaging of the systemic recruitment of fibroblasts to the angiogenic rim of ovarian carcinoma tumors. AB - Tumor-associated stroma, in general, and tumor fibroblasts and myofibroblasts, in particular, play a role in tumor progression. We previously reported that myofibroblast infiltration into implanted ovarian carcinoma spheroids marked the exit of tumors from dormancy and that these cells contributed to vascular stabilization in ovarian tumors by expression of angiopoietin-1 and angiopoietin 2. Ex vivo labeling of fibroblasts with either magnetic resonance or optical probes rendered them detectable for in vivo imaging. Thus, magnetic resonance imaging (MRI) follow-up was feasible by biotin-bovine serum albumin-gadolinium diethylenetriaminepentaacetic acid or iron oxide particles, whereas labeling with near-IR and fluorescent vital stains enabled in vivo visualization by near-IR imaging and two-photon microscopy. Using this approach, we show here that prelabeled fibroblasts given i.p. to CD-1 nude mice can be followed in vivo by MRI and optical imaging over several days, revealing their extensive recruitment into the stroma of remote s.c. MLS human epithelial ovarian carcinoma tumors. Two photon microscopy revealed the alignment of these invading fibroblasts in the outer rim of the tumor, colocalizing with the angiogenic neovasculature. Such angiogenic vessels remained confined to the stroma tracks within the tumor and did not penetrate the tumor nodules. These results provide dynamic evidence for the role of tumor fibroblasts in maintenance of functional tumor vasculature and offer means for image-guided targeting of these abundant stroma cells to the tumor as a possible mechanism for cellular cancer therapy. PMID- 17909024 TI - An increased expression of cysteinyl leukotriene 2 receptor in colorectal adenocarcinomas correlates with high differentiation. AB - Increased levels of inflammatory mediators such as cysteinyl leukotrienes (CysLT) have been found in and around tumors. These data, along with our previous observation that the G-protein-coupled receptor CysLT(1)R, which signals survival and proliferation, is up-regulated in colon cancer, suggest an important role for CysLT(1)R in tumor development. The objective of this study was to examine the expression and function of the low-affinity CysLT2 receptor (CysLT2R) in colon cancer. We found lower expression levels of CysLT2R compared with CysLT(1)R in cancer cell lines as well as clinical tumor material. Interestingly, CysLT2R, like CysLT(1)R, was found to be one of few G-protein-coupled receptors that are located both at the plasma membrane and the nuclear membrane. No effect of CysLT2R signaling on cell proliferation was observed, nor was there a correlation between CysLT2R and different proliferation markers such as Ki-67 and cyclooxygenase-2 in the tumor material. Instead, we found that activation of this receptor in colon cancer cells led to cellular differentiation similar to the effects of butyrate treatment. In accordance with this finding, we found that reduced expression of CysLT2R in colon cancer was associated with poor prognosis. We report the novel finding that CysLT2R signaling leads to terminal differentiation of colon carcinoma cells and growth inhibition, and that its expression is relatively high in less malignant forms of colon cancer. These data suggest that the balance between these two receptors is important for tumor progression and disease outcome. PMID- 17909026 TI - Snail-induced down-regulation of DeltaNp63alpha acquires invasive phenotype of human squamous cell carcinoma. AB - p63 is a member of the p53 family and regulates crucial events in the formation of epithelial structures, but the role of p63 in tumor is unclear. We found that Snail-induced epithelial-to-mesenchymal transition (EMT) is accompanied by down regulation of p63 in human squamous cell carcinomas (SCC). DeltaNp63alpha is the predominantly expressed p63 isoform in SCC cells. DeltaNp63 promoter activity required a CAAT/enhancer binding protein (C/EBP) binding element and was reduced remarkably by Snail. Down-regulation of DeltaNp63alpha and reduction of C/EBPalpha were observed in EMT phenotype cells, which exhibited invasive activity in vitro. p63 knockdown in cells enhanced invasive activity in the presence of E-cadherin. Conversely, forced expression of DeltaNp63alpha blocked invasive activity of cells with the EMT phenotype. These findings indicate that Snail down-regulates DeltaNp63alpha, leading to acquisition of the invasive phenotype by SCC. The invasive activity caused by down-regulation of DeltaNp63alpha does not require down-regulation of E-cadherin. PMID- 17909025 TI - Roles for the stem cell associated intermediate filament Nestin in prostate cancer migration and metastasis. AB - The intermediate filament protein Nestin identifies stem/progenitor cells in adult tissues, but the function of Nestin is poorly understood. We investigated Nestin expression and function in common lethal cancers. Nestin mRNA was detected in cell lines from small cell lung, and breast cancers, and particularly elevated in cell lines derived from prostate cancer metastases. Whereas the androgen independent lines PC3, 22RV1, and DU145 all expressed Nestin transcripts under standard culture conditions, the androgen-dependent line LnCaP expressed Nestin only on androgen withdrawal. We confirmed associations of Nestin expression, androgen withdrawal, and metastatic potential by immunohistochemical analysis of samples from 254 prostate cancer patients. Cytoplasmic Nestin protein was readily identifiable in prostate cancer cells from 75% of patients with lethal androgen independent disease, even in cancer sampled from the prostate itself. However, Nestin expression was undetectable in localized androgen-deprived tumors and in metastases without prior androgen deprivation. To address its function, we reduced Nestin levels with short hairpin RNAs, markedly inhibiting in vitro migration and invasion in prostate cancer cells but leaving cell growth intact. Nestin knockdown also diminished metastases 5-fold compared with controls despite uncompromised tumorigenicity at the site of inoculation. These results specify a function for Nestin in cell motility and identify a novel pathway for prostate cancer metastasis. Activity of this pathway may be selected by the extraprostatic environment or, as supported by our data, may originate within the prostate after androgen deprivation. Further dissection of this novel Nestin migration pathway may lead to strategies to prevent and neutralize metastatic spread. PMID- 17909028 TI - Whole-genome profiling in liposarcomas reveals genetic alterations common to specific telomere maintenance mechanisms. AB - Telomere attrition ultimately leads to the activation of protective cellular responses, such as apoptosis or senescence. Impairment of such mechanisms can allow continued proliferation despite the presence of dysfunctional telomeres. Under such conditions, high levels of genome instability are often engendered. Data from both mouse and human model systems indicate that a period of genome instability might facilitate tumorigenesis. Here, we use a liposarcoma model system to assay telomere maintenance mechanism (TMM)-specific genetic alterations. A multiassay approach was used to assess the TMMs active in tumors. Genomic DNA from these samples was then analyzed by high-resolution DNA mapping array to identify genetic alterations. Our data reveal a higher level of genome instability in alternative lengthening of telomere (ALT)-positive tumors compared with telomerase-positive tumors, whereas tumors lacking both mechanisms have relatively low levels of genome instability. The bulk of the genetic changes are amplifications, regardless of the mode of telomere maintenance used. We also identified genetic changes specific to the ALT mechanism (e.g., deletion of chromosome 1q32.2-q44) as well as changes that are underrepresented among ALT positive tumors, such as amplification of chromosome 12q14.3-q21.2. Taken together, these studies provide insight into the molecular pathways involved in the regulation of ALT and reveal several loci that might be exploited either as prognostic markers or targets of chemotherapeutic intervention. PMID- 17909027 TI - Signal transducer and activator of transcription 1 regulates both cytotoxic and prosurvival functions in tumor cells. AB - Elsewhere, we reported that multiple serial in vivo passage of a squamous cell carcinoma cells (SCC61) concurrent with ionizing radiation (IR) treatment resulted in the selection of radioresistant tumor (nu61) that overexpresses the signal transducer and activator of transcription 1 (Stat1)/IFN-dependent pathway. Here, we report that (a) the Stat1 pathway is induced by IR, (b) constitutive overexpression of Stat1 is linked with failure to transmit a cytotoxic signal by radiation or IFNs, (c) selection of parental cell line SCC61 against IFN-alpha and IFN-gamma leads to the same IR- and IFN-resistant phenotype as was obtained by IR selection, and (d) suppression of Stat1 by short hairpin RNA renders the IR resistant nu61 cells radiosensitive to IR. We propose a model that transient induction of Stat1 by IFN, IR, or other stress signals activates cytotoxic genes and cytotoxic response. Constitutive overexpression of Stat1 on the other hand leads to the suppression of the cytotoxic response and induces prosurvival genes that, at high levels of Stat1, render the cells resistant to IR or other inducers of cell death. PMID- 17909030 TI - Relocalized p27Kip1 tumor suppressor functions as a cytoplasmic metastatic oncogene in melanoma. AB - The p27 tumor suppressor negatively regulates G1 cell cycle progression. However, human malignancies rarely select for deletion/inactivation of p27, a hallmark of tumor suppressor genes. Instead, p27 is degraded or relocalized to the cytoplasm in aggressive malignancies, supporting the notion that p27 sequestration from its nuclear cyclin:cyclin-dependent kinase (cdk) targets is critical. However, emerging cell biology data suggest a novel cdk-independent cytoplasmic function of p27 in cell migration. Here, we find cytoplasmic p27 in 70% of invasive and metastatic melanomas. In contrast, no cytoplasmic p27 was detected in noninvasive, basement membrane-confined melanoma in situ, suggesting a late oncogenic role for cytoplasmic p27 in metastasis. Targeted cytoplasmic expression of wild-type or non-cdk-binding p27 at subphysiologic levels induced melanoma motility and resulted in numerous metastases to lymph node, lung, and peritoneum. These observations point to a prominent role of cytoplasmic p27 in metastatic disease that is independent of cyclin:cdk regulation or mere nuclear loss. PMID- 17909029 TI - The phosphoinositide kinase PIKfyve mediates epidermal growth factor receptor trafficking to the nucleus. AB - ErbB receptor tyrosine kinases can transit to nuclei in tumor cells, where they have been shown to regulate gene expression as components of transcriptional complexes. Quantitative analysis of a human bladder cancer tissue microarray identified nuclear epidermal growth factor receptor (EGFR) in tumor cells and also showed an increased frequency of this histologic feature in cancer relative to normal tissues. This observation suggests a potential role for nuclear EGFR in bladder cancer. We confirmed that EGFR could be induced to transit to nuclei in cultured human bladder cancer cells in response to the urothelial cell growth factor and EGFR ligand heparin-binding EGF-like growth factor (HB-EGF). Mass spectrometric analysis of EGFR immune complexes from a transitional carcinoma cell line (TCCSUP) identified the phosphoinositide kinase, PIKfyve, as a potential component of the EGFR trafficking mechanism. RNA silencing indicated that PIKfyve is a mediator of HB-EGF-stimulated EGFR nuclear trafficking, EGFR binding to the cyclin D1 promoter, and cell cycle progression. These results identify a novel mediator of the EGFR transcription function and further suggest that nuclear EGFR and the lipid kinase PIKfyve may play a role in bladder oncogenesis. PMID- 17909031 TI - In vivo significance of the G2 restriction point. AB - Loss of activity of the retinoblastoma pathway is a common event in human cancer. Mouse models have revealed that tumorigenesis by loss of Rb was accelerated by concomitant loss of the cell cycle inhibitor p27KIP1. This has been attributed to reduced apoptosis and weakening of the G1 checkpoint. However, the role of p27KIP1 in a recently identified G2 restriction point may offer an alternative explanation for this synergy. Here, we have investigated the significance of the G2 restriction point in Rb-deficient pituitaries. We show that Rb loss in the pituitary gland activated the G2 restriction point, as evidenced by the appearance of cyclin B1-p27KIP1 complexes. Somewhat unexpectedly, these complexes remained present in Rb-deficient tumors. These results indicate that the G2 restriction point does operate in vivo. However, in the pituitary gland, this mechanism seems to retard rather than to prevent tumor growth. PMID- 17909033 TI - Inhibition of the proteasome activity by gallium(III) complexes contributes to their anti prostate tumor effects. AB - The investigation of metal-based complexes with potential antitumor activity has been of paramount importance in recent years due to the successful use of cisplatin against various cancers. Gallium(III) and subsequently developed gallium(III)-containing complexes have shown promising antineoplastic effects when tested in a host of malignancies, specifically in lymphomas and bladder cancer. However, the molecular mechanism responsible for their anticancer effect is yet to be fully understood. We report here for the first time that the proteasome is a molecular target for gallium complexes in a variety of prostate cancer cell lines and in human prostate cancer xenografts. We tested five gallium complexes (1-5) in which the gallium ion is bound to an NN'O asymmetrical ligand containing pyridine and substituted phenolate moieties in a 1:2 (M/L) ratio. We found that complex 5 showed superior proteasome inhibitory activity against both 26S proteasome (IC50, 17 micromol/L) and purified 20S (IC50, 16 micromol/L) proteasome. Consistently, this effect was associated with apoptosis induction in prostate cancer cells. Additionally, complex 5 was able to exert the same effect in vivo by inhibiting growth of PC-3 xenografts in mice (66%), which was associated with proteasome inhibition and apoptosis induction. Our results strongly suggest that gallium complexes, acting as potent proteasome inhibitors, have a great potential to be developed into novel anticancer drugs. PMID- 17909032 TI - Glutathione transferase pi plays a critical role in the development of lung carcinogenesis following exposure to tobacco-related carcinogens and urethane. AB - Human cancer is controlled by a complex interaction between genetic and environmental factors. Such environmental factors are well defined for smoking induced lung cancer; however, the roles of specific genes have still to be elucidated. Glutathione transferase pi (GSTP) catalyzes the detoxification of electrophilic diol epoxides produced by the metabolism of polycyclic aromatic hydrocarbons such as benzo[a]pyrene (BaP), a common constituent of tobacco smoke. Activity-altering polymorphisms in Gstp have therefore been speculated to be potential risk modifiers in lung cancer development. To clearly establish a role for GSTP in lung tumorigenesis, we investigated whether deletion of the murine Gstp genes (Gstp1 and Gstp2) alters susceptibility to chemically induced lung tumors following exposure to BaP, 3-methylcholanthrene (3-MC), and urethane. Gstp null mice were found to have substantially increased numbers of adenomas relative to wild-type mice following exposure to all three compounds (8.3-, 4.3-, and 8.7 fold increase for BaP, 3-MC, and urethane, respectively). In Gstp-null mice, the capacity of pulmonary cytosol to catalyze conjugation of the BaP diol epoxide was significantly reduced. Concomitant with this, a significant increase in the level of BaP DNA adducts was measured in the lungs of null animals; however, no increase in DNA adducts was measured in the case of 3-MC exposure, suggesting that an alternative protective pathway exists. Indeed, significant differences in pulmonary gene expression profiles were also noted between wild-type and null mice. This is the first report to establish a clear correlation between Gstp status and lung cancer in vivo. PMID- 17909034 TI - Insulin receptor substrate-1 is an important mediator of ovarian cancer cell growth suppression by all-trans retinoic acid. AB - There is a need to identify more effective drugs for the treatment of ovarian cancer as it is the leading cause of death among gynecologic tumors. All-trans retinoic acid (ATRA), a natural retinoid, arrests the growth of CA-OV3 ovarian carcinoma cells in G(0)-G(1). Because the insulin-like growth factor-I receptor has been implicated in the proliferation of various tumors, we investigated its potential role in the suppression of ovarian cancer cell growth by ATRA. Our studies revealed that insulin receptor substrate-1 (IRS-1) protein levels decrease in CA-OV3 cells on ATRA treatment, whereas no differences in IRS-1 levels were seen in the ATRA-resistant SK-OV3 cells. Moreover, CA-OV3 clones overexpressing IRS-1 were growth inhibited less by ATRA, whereas SK-OV3 clones in which levels of IRS-1 were reduced by expression of antisense IRS-1 became sensitive to growth inhibition by ATRA treatment. Studies to determine the mechanism by which ATRA reduced IRS-1 expression showed that ATRA altered steady state levels of IRS-1 mRNA and the stability of IRS-1 protein. Finally, the role of IRS-1 as a potential molecular target of ATRA in ovarian tumors was assessed by immunohistochemistry in an ovarian cancer tissue array. Compared with normal ovary, the majority of malignant epithelial ovarian tumors overexpressed IRS-1. Thus, there seems to be a correlation between IRS-1 expression and malignancy in ovarian tumors. Our results suggest that IRS-1 is in fact an important growth regulatory molecule that can be a potential effective target for chemotherapeutic intervention with growth-suppressive agents, including retinoids. PMID- 17909035 TI - Cancer resistance in transgenic mice expressing the SAC module of Par-4. AB - Prostate apoptosis response-4 (Par-4) is a tumor-suppressor protein that induces apoptosis in cancer cells, but not in normal/immortalized cells. The cancer specific proapoptotic action of Par-4 is encoded in its centrally located SAC domain. We report here the characterization of a novel mouse model with ubiquitous expression of the SAC domain. Although SAC transgenic mice displayed normal development and life span, they were resistant to the growth of spontaneous, as well as oncogene-induced, autochthonous tumors. Resistance to tumorigenesis was linked to inhibition of nuclear factor-kappaB activity and induction of apoptosis by the SAC domain. Collectively, our findings provide genetic evidence that the SAC domain of Par-4 confers cancer resistance in transgenic mice without compromising normal viability or aging, and may have therapeutic significance. PMID- 17909038 TI - Aberrant expression of cortactin in head and neck squamous cell carcinoma cells is associated with enhanced cell proliferation and resistance to the epidermal growth factor receptor inhibitor gefitinib. AB - The CTTN gene (formerly designated EMS1), encodes cortactin, a key regulator of dynamic actin networks. Both CTTN and CCND1, the latter encoding the cell cycle regulator cyclin D1, reside at chromosomal locus 11q13, a region commonly amplified in breast cancers and head and neck squamous cell carcinoma (HNSCC). Previously, we identified a novel role for cortactin in cancer cells, whereby cortactin overexpression attenuated ligand-induced down-regulation of the epidermal growth factor (EGF) receptor (EGFR), leading to sustained signaling. However, how this affected growth factor-induced cellular responses was unclear. Here, by modulation of cortactin expression in a panel of HNSCC cell lines, we show that cortactin overexpression enhances serum- and EGF-stimulated proliferation under both anchorage-dependent and anchorage-independent conditions and also increases resistance to anoikis (detachment-induced apoptosis). These effects are associated with increased activation of extracellular signal regulated kinase and/or AKT. Furthermore, we report that cortactin stabilizes the c-MET receptor tyrosine kinase and enhances hepatocyte growth factor-induced mitogenesis and cell scattering. Therefore, cortactin may modulate signaling by a broader range of receptors than originally proposed and thereby affect a variety of responses. Finally, we have determined that cortactin overexpression, either alone or in combination with cyclin D1 up-regulation, promotes resistance to the EGFR kinase inhibitor gefitinib. These findings indicate that cortactin may play multiple roles in progression of HNSCC and should be evaluated as a marker of prognosis, disease progression, and therapeutic responsiveness, particularly to EGFR-directed agents. PMID- 17909037 TI - Peroxiredoxin 1 interacts with androgen receptor and enhances its transactivation. AB - Although hypoxia is accepted as an important microenvironmental factor influencing tumor progression and treatment response, it is usually regarded as a static global phenomenon. Consequently, less attention is given to the impact of dynamic changes in tumor oxygenation in regulating the behavior of cancer cells. Androgen receptor (AR) signaling plays a critical role in prostate cancer. We previously reported that hypoxia/reoxygenation, an in vitro condition used to mimic an unstable oxygenation climate in a tumor, stimulates AR activation. In the present study, we showed that peroxiredoxin 1 (Prx1), a member of the peroxiredoxin protein family, acts as a key mediator in this process. We found that the aggressive LN3, C4-2, and C4-2B prostate cancer cell lines derived from LNCaP possess constitutively elevated Prx1 compared with parental cells, and display greater AR activation in response to hypoxia/reoxygenation. Although the cell survival-enhancing property of Prx1 has traditionally been attributed to its antioxidant activity, the reactive oxygen species-scavenging activity of Prx1 was not essential for AR stimulation because Prx1 itself was oxidized and inactivated by hypoxia/reoxygenation. Increased AR transactivation was observed when wild type Prx1 or mutant Prx1 (C52S) lacking antioxidant activity was introduced into LNCaP cells. Reciprocal immunoprecipitation, chromatin immunoprecipitation, and in vitro pull-down assays corroborated that Prx1 interacts with AR and enhances its transactivation. We also show that Prx1 is capable of sensitizing a ligand stimulated AR. Based on the above information, we suggest that disrupting the interaction between Prx1 and AR may serve as a fruitful new target in the management of prostate cancer. PMID- 17909036 TI - Nitric oxide inactivates the retinoblastoma pathway in chronic inflammation. AB - Patients with chronic inflammatory bowel disease have a high risk of colon cancer. The molecules that initiate and promote colon cancer and the cancer pathways altered remain undefined. Here, using in vitro models and a mouse model of colitis, we show that nitric oxide (NO) species induce retinoblastoma protein (pRb) hyperphosphorylation and inactivation, resulting in increased proliferation through the pRb-E2F1 pathway. NO-driven pRb hyperphosphorylation occurs through soluble guanylyl cyclase/guanosine 3',5'-cyclic monophosphate signaling and is dependent on the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase MEK/ERK and phosphatidylinositol 3-kinase/AKT pathways. Our results reveal a link between NO and pRb inactivation and provide insight into molecules that can be targeted in the prevention of the inflammation-to-cancer sequence. PMID- 17909039 TI - beta-catenin regulates multiple steps of RNA metabolism as revealed by the RNA aptamer in colon cancer cells. AB - Nuclear beta-catenin forms a transcription complex with TCF-4, which is implicated in colon cancer development and progression. Recently, we and others have shown that beta-catenin could be a regulator of RNA splicing and it also stabilizes the cyclooxygenase-2 (COX-2) mRNA. Here, we further explored the role of beta-catenin in the RNA metabolism in colon cancer cells. To specifically modulate the subcellular functions of beta-catenin, we expressed the RNA aptamer in the form of RNA intramers with unique cellular localizations. The nucleus expressed RNA intramer proved to be effective in reducing the protein-protein interaction between beta-catenin and TCF-4, thus shown to be a specific regulator of beta-catenin-activated transcription. It could also regulate the alternative splicing of E1A minigene in diverse colon cancer cell lines. In addition, we tested whether beta-catenin could stabilize any other mRNAs and found that cyclin D1 mRNA was also bound and stabilized by beta-catenin. Significantly, the cytoplasm-expressed RNA intramer reverted the beta-catenin-induced COX-2 and cyclin D1 mRNA stabilization. We show here that beta-catenin regulated multiple steps of RNA metabolism in colon cancer cells and might be the protein factor coordinating RNA metabolism. We suggest that the RNA intramers could provide useful ways for inhibiting beta-catenin-mediated transcription and RNA metabolism, which might further enhance the antitumorigenic effects of these molecules in colon cancer cells. PMID- 17909041 TI - Association of Wwox with ErbB4 in breast cancer. AB - WWOX, WW domain-containing oxidoreductase, is a tumor suppressor that is altered in many human cancers, including breast cancer. Wwox interacts with the ErbB4 receptor, reduces nuclear translocation of the cleaved intracellular domain of ErbB4, and inhibits its transactivation function mediated through Yes-associated protein. Here, we assessed the clinical significance of the Wwox-ErbB4 association. We determined Wwox protein expression by immunohistochemistry in a series of 556 breast cancers. Wwox expression was absent in 36% of the cancers, and loss of Wwox expression was associated with unfavorable outcome (P = 0.02). Membranous location of ErbB4 was associated with favorable survival compared with women whose cancer lacked such ErbB4 expression (P = 0.02). Wwox expression was strongly associated with membranous ErbB4 localization (P = 0.0003) and with overall ErbB4 expression (P = 0.0002). Coexpression of membranous ErbB4 and Wwox was associated with favorable outcome compared with cases with membranous ErbB4 and no Wwox immunoreactivity (P = 0.002). In vitro, Wwox associated with the two ErbB4 isoforms, JM-a CYT-1 and JM-a CYT-2, expressed in breast cancer. Moreover, expression of Wwox both in vitro and in vivo led to accumulation of total full length membrane-associated ErbB4. These results suggest that expression of Wwox is associated with ErbB4 expression and that their coexpression has prognostic significance in breast cancer. PMID- 17909040 TI - The alternative reading frame tumor suppressor antagonizes hypoxia-induced cancer cell migration via interaction with the COOH-terminal binding protein corepressor. AB - The alternative reading frame (ARF) tumor suppressor exerts both p53-dependent and p53-independent activities critical to the prevention of cancer in mice and humans. Recent evidence from mouse models suggests that when p53 is absent, further loss of ARF can widen the tumor spectrum, and potentiate invasion and metastasis. A major target of the p53-independent activity of ARF is the COOH terminal binding protein (CtBP) family of metabolically regulated transcriptional corepressors, which are degraded upon acute exposure to the ARF protein. CtBPs are activated under conditions of metabolic stress, such as hypoxia, to repress epithelial and proapoptotic genes, and can mediate hypoxia-induced migration of cancer cells. The possibility that ARF could suppress tumor cell migration as part of its p53-independent activities was thus explored. Small-interfering RNA (siRNA)-mediated knockdown of ARF in human lung carcinoma cells led to increased cell migration, especially during hypoxia, and this effect was blocked by concomitant treatment with CtBP2 siRNA. Introduction of ARF into p53 and ARF-null human colon cancer cells inhibited hypoxia-induced migration. Furthermore, overexpression of CtBP2 in ARF-expressing cells enhanced cell migration, and an ARF mutant defective in CtBP-family binding was impaired in its ability to inhibit cell migration induced by CtBP2. ARF depletion or CtBP2 overexpression was associated with decreased PTEN expression and activation of the phosphatidylinositol 3-kinase pathway, and a phosphatidylinositol 3-kinase inhibitor blocked CtBP2-mediated cell migration. Thus, ARF can suppress cell migration by antagonizing CtBP2 and the phosphatidylinositol 3-kinase pathway, and these data may explain the increased aggressiveness of ARF-null tumors in mouse models. PMID- 17909042 TI - Antitumor and antivascular effects of AVE8062 in ovarian carcinoma. AB - The purpose of this study was to examine the therapeutic efficacy and underlying mechanisms of action of a vascular-disrupting agent, AVE8062, and to determine its effects on tumor metabolic activity. The in vitro and in vivo effects of AVE8062 alone and in combination with docetaxel were tested in chemotherapy sensitive and chemotherapy-resistant ovarian cancer models. Tumors were analyzed for necrosis, microvessel density, endothelial cell apoptosis, and proliferation following treatment. The effect of AVE8062 on tumor regression and metabolic activity was examined by magnetic resonance (MR) or by [18F]fluorodeoxyglucose ([18F]FDG) uptake by positron emission tomography (PET) with MR imaging, respectively. AVE8062 monotherapy was effective in inhibiting tumor growth in all models (range 43-51% versus control; P < 0.05). Combination therapy was even more effective in inhibiting tumor growth (range 76-90% compared with controls, P < 0.01). AVE8062 in combination with chemotherapy significantly prolonged survival in HeyA8-injected mice (P < 0.001) compared with other groups. AVE8062-based therapy resulted in rapid development of central tumor necrosis, decreased microvessel density, decreased proliferation, and induction of apoptosis of tumor associated endothelial cells. MR imaging showed regression of established HeyA8 ovarian tumors and [18F]FDG PET with MR showed rapid decrease in metabolic activity after AVE8062 therapy. Combination of AVE8062 plus docetaxel results in potent inhibition of ovarian cancer growth. These results suggest that AVE8062 may be useful as a clinical therapeutic approach for ovarian cancer patients and that functional [18F]FDG PET imaging may predict clinical response before an anatomic reduction in tumor size. PMID- 17909043 TI - The activity of zoledronic Acid on neuroblastoma bone metastasis involves inhibition of osteoclasts and tumor cell survival and proliferation. AB - Metastasis to the bone is seen in 56% of patients with neuroblastoma and contributes to morbidity and mortality. Using a murine model of bone invasion, we have reported previously that neuroblastoma cells invade the bone by activating osteoclasts. Here, we investigated the antitumoral and antiosteolytic activities of zoledronic acid, a bisphosphonate inhibitor of osteoclasts, in combination with cytotoxic chemotherapy in our model. We first show that zoledronic acid given at the same time (early prevention) or 2 weeks after tumor cell injection (late prevention) significantly prevented the formation of severe osteolytic lesions. It also prevented formation of these lesions when given 4 weeks after tumor cell injection (intervention) when combined with chemotherapy including cyclophosphamide and topotecan. The combination of zoledronic acid + cyclophosphamide/topotecan also significantly improved survival (P < 0.001). In mice treated with zoledronic acid, we observed a marked inhibition of osteoclasts inside the bone associated with a decrease in tumor cell proliferation and increase in tumor cell apoptosis. In vitro, zoledronic acid inhibited neuroblastoma cell proliferation and induced apoptosis, and these effects were significantly enhanced by the addition of 4-hydroxyperoxycyclophosphamide (4-HC). The proapoptotic effect of zoledronic acid and zoledronic acid in combination with 4-HC on tumor cells was associated with an increase in caspase-3 activity and a decrease in phosphorylated Bcl-2, Bcl-2, and Bcl-X(L) expression. Zoledronic acid inhibited the association of Ras with the plasma membrane and activation of c-Raf, Akt, and extracellular signal-regulated kinase 1/2. The data indicate that zoledronic acid, in addition to inhibiting osteoclasts, is active against tumor cells and suggest that zoledronic acid in combination with cytotoxic chemotherapy may be effective in children with neuroblastoma that has metastasized to the bone. PMID- 17909044 TI - Class III beta-tubulin mediates sensitivity to chemotherapeutic drugs in non small cell lung cancer. AB - First line therapy for non-small cell lung carcinoma (NSCLC) commonly includes combination therapy with a tubulin-binding agent (TBA) and a DNA-damaging agent. TBAs suppress microtubule dynamics by binding to the beta-tubulin subunit of alpha/beta-tubulin, inducing mitotic arrest and apoptosis. Up-regulation of class III beta-tubulin (betaIII-tubulin) has been implicated in clinical resistance in NSCLC, ovarian and breast tumors treated in combination with a TBA and DNA damaging agent. To investigate the functional significance of betaIII-tubulin in resistance to both these classes of agents, small interfering RNA (siRNA) was used to silence the expression of this isotype in two NSCLC cell lines, NCI-H460 and Calu-6. Reverse transcription-PCR and immunoblotting showed that betaIII siRNA potently inhibited the expression of betaIII-tubulin, without affecting the expression of other major beta-tubulin isotypes. Clonogenic assays showed that betaIII-siRNA cells were significantly more sensitive to TBAs, paclitaxel, vincristine, and vinorelbine, and for the first time, DNA-damaging agents, cisplatin, doxorubicin, and etoposide compared with controls. Cell cycle analysis of H460 betaIII-siRNA cells showed reduced accumulation at the G(2)-M boundary and an increase in the sub-G(1) population in response to TBA treatment compared with control cells. Importantly, betaIII-siRNA cells displayed a significant dose dependent increase in Annexin V staining when treated with either paclitaxel or cisplatin, compared with controls. These findings have revealed a novel role for betaIII-tubulin in mediating response to both TBA and DNA-damaging agent therapy and may have important implications for improving the targeting and treatment of drug-refractory NSCLC. PMID- 17909045 TI - Serial assessment of human tumor burdens in mice by the analysis of circulating DNA. AB - Internal human xenografts provide valuable animal models to study the microenvironments and metastatic processes occurring in human cancers. However, the use of such models is hampered by the logistical difficulties of reproducibly and simply assessing tumor burden. We developed a high-sensitivity assay for quantifying human DNA in small volumes of mouse plasma, enabling in-life monitoring of systemic tumor burden. Growth kinetics analyses of various xenograft models showed the utility of circulating human DNA as a biomarker. We found that human DNA concentration reproducibly increased with disease progression and decreased after successful therapeutic intervention. A marked, transient spike in circulating human tumor DNA occurred immediately after cytotoxic therapy or surgery. This simple assay may find broad utility in target validation studies and preclinical drug development programs. PMID- 17909046 TI - Systemic therapy of spontaneous prostate cancer in transgenic mice with oncolytic herpes simplex viruses. AB - Oncolytic viruses are an innovative therapeutic strategy for cancer, wherein viral replication and cytotoxicity are selective for tumor cells. Here we show the efficacy of systemically administered oncolytic viruses for the treatment of spontaneously arising tumors, specifically the use of oncolytic herpes simplex viruses (HSV) administered i.v. to treat spontaneously developing primary and metastatic prostate cancer in the transgenic TRAMP mouse, which recapitulates human prostate cancer progression. Four administrations of systemically delivered NV1023 virus, an HSV-1/HSV-2 oncolytic recombinant, to TRAMP mice at 12 or 18 weeks of age (presence of prostate adenocarcinoma or metastatic disease, respectively) inhibited primary tumor growth and metastases to lymph nodes. Expression of interleukin 12 (IL-12) from NV1042 virus, a derivative of NV1023, was additionally effective, significantly reducing the frequency of development of prostate cancer and lung metastases, even when the mice were treated after the onset of metastasis at 18 weeks of age. NV1042-infected cells, as detected by 5 bromo-4-chloro-3-indolyl-beta-d-galactopyranoside staining for Lac Z expressed by the virus, were present in prostate tumors 1 week after the final virus injection and viral DNA was detected at 2 weeks after final virus injection by real-time PCR in primary and metastatic tumors but not in liver or blood. No toxicity was observed in any of the treated mice. The efficacy of the IL-12-expressing NV1042 virus in this aggressive prostate cancer model using a clinically relevant treatment paradigm merits its consideration for clinical studies. PMID- 17909047 TI - Targeting cyclooxygenase-2 and the epidermal growth factor receptor for the prevention and treatment of intestinal cancer. AB - Clinical and animal studies indicate a role for cyclooxygenase-2 (COX-2) and the epidermal growth factor receptor (EGFR) in the development and progression of intestinal polyps and cancers. Although this combination of enzyme inhibition has shown synergy in intestinal polyp and tumor models, the exact mechanism for these effects remains undefined. Therefore, we sought to define the molecular mechanisms through which this process occurs. We observed a significant reduction in the number and size of small intestinal polyps in APC(min+/-) mice treated with either celecoxib (a selective COX-2 inhibitor) or erlotinib (Tarceva, an EGFR inhibitor). However, in combination, there was an overall prevention in the formation of polyps by over 96%. Furthermore, we observed a 70% reduction of colorectal xenograft tumors in mice treated with the combination and microarray analysis revealed genes involved in cell cycle progression were negatively regulated. Although we did not observe significant changes in mRNAs of genes with known apoptotic function, there was a significant increase of apoptosis in tumors from animals treated with the combination. The inhibition of EGFR also induced the down-regulation of COX-2 and further inhibited prostaglandin E2 formation. We observed similar effects on the prevention of intestinal adenomas and reduction of xenograft tumor volume when nonselective COX inhibitors were used in combination with erlotinib. Together, these findings suggest that the inhibition of both COX-2 and EGFR may provide a better therapeutic strategy than either single agent through a combination of decreased cellular proliferation and prostaglandin signaling as well as increased apoptosis. PMID- 17909048 TI - ABCG2/BCRP expression modulates D-Luciferin based bioluminescence imaging. AB - Bioluminescence imaging (BLI) is becoming indispensable to the study of transgene expression during development and, in many in vivo models of disease such as cancer, for high throughput drug screening in vitro. Because reaction of d luciferin with firefly luciferase (fLuc) produces photons of sufficiently long wavelength to permit imaging in intact animals, use of this substrate and enzyme pair has become the method of choice for performing BLI in vivo. We now show that expression of the ATP-binding cassette (ABC) family transporter ABCG2/BCRP affects BLI signal output from the substrate d-luciferin. In vitro studies show that d-luciferin is a substrate for ABCG2/BCRP but not for the MDR1 P glycoprotein (ABCB1/Pgp), multidrug resistance protein 1 (MRP1/ABCC1), or multidrug resistance protein 2 (MRP2/ABCC2). d-Luciferin uptake within cells is shown to be modulated by ABC transporter inhibitors, including the potent and selective ABCG2/BCRP inhibitor fumitremorgin C. Images of xenografts engineered to express transgenic ABCG2/BCRP, as well as xenografts derived from the human prostate cancer cell line 22Rv1 that naturally express ABCG2/BCRP, show that ABCG2/BCRP expression and function within regions of interest substantially influence d-luciferin-dependent bioluminescent output in vivo. These findings highlight the need to consider ABCG2/BCRP effects during d-luciferin-based BLI and suggest novel high throughput methods for identifying new ABCG2/BCRP inhibitors. PMID- 17909049 TI - Depletion of peripheral macrophages and brain microglia increases brain tumor titers of oncolytic viruses. AB - Clinical trials have proven oncolytic virotherapy to be safe but not effective. We have shown that oncolytic viruses (OV) injected into intracranial gliomas established in rodents are rapidly cleared, and this is associated with up regulation of markers (CD68 and CD163) of cells of monocytic lineage (monocytes/microglia/macrophages). However, it is unclear whether these cells directly impede intratumoral persistence of OV through phagocytosis and whether they infiltrate the tumor from the blood or the brain parenchyma. To investigate this, we depleted phagocytes with clodronate liposomes (CL) in vivo through systemic delivery and ex vivo in brain slice models with gliomas. Interestingly, systemic CL depleted over 80% of peripheral CD163+ macrophages in animal spleen and peripheral blood, thereby decreasing intratumoral infiltration of these cells, but CD68+ cells were unchanged. Intratumoral viral titers increased 5 fold. In contrast, ex vivo CL depleted only CD68+ cells from brain slices, and intratumoral viral titers increased 10-fold. These data indicate that phagocytosis by both peripheral CD163+ and brain-resident CD68+ cells infiltrating tumor directly affects viral clearance from tumor. Thus, improved therapeutic efficacy may require modulation of these innate immune cells. In support of this new therapeutic paradigm, we observed intratumoral up-regulation of CD68+ and CD163+ cells following treatment with OV in a patient with glioblastoma. PMID- 17909050 TI - Triptolide induces pancreatic cancer cell death via inhibition of heat shock protein 70. AB - Pancreatic cancer is highly resistant to current chemotherapy agents. We therefore examined the effects of triptolide (a diterpenoid triepoxide) on pancreatic cancer growth and local-regional tumor spread using an orthotopic model of pancreatic cancer. We have recently shown that an increased level of HSP70 in pancreatic cancer cells confers resistance to apoptosis and that inhibiting HSP70 induces apoptosis in these cells. In addition, triptolide was recently identified as part of a small molecule screen, as a regulator of the human heat shock response. Therefore, our aims were to examine the effects of triptolide on (a) pancreatic cancer cells by assessing viability and apoptosis, (b) pancreatic cancer growth and local invasion in vivo, and (c) HSP70 levels in pancreatic cancer cells. Incubation of PANC-1 and MiaPaCa-2 cells with triptolide (50-200 nmol/L) significantly reduced cell viability, but had no effect on the viability of normal pancreatic ductal cells. Triptolide induced apoptosis (assessed by Annexin V, caspase-3, and terminal nucleotidyl transferase-mediated nick end labeling) and decreased HSP70 mRNA and protein levels in both cell lines. Triptolide (0.2 mg/kg/d for 60 days) administered in vivo decreased pancreatic cancer growth and significantly decreased local-regional tumor spread. The control group of mice had extensive local invasion into adjacent organs, including the spleen, liver, kidney, and small intestine. Triptolide causes pancreatic cancer cell death in vitro and in vivo by induction of apoptosis and its mechanism of action is mediated via the inhibition of HSP70. Triptolide is a potential therapeutic agent that can be used to prevent the progression and metastases of pancreatic cancer. PMID- 17909051 TI - Targeting CCL2 with systemic delivery of neutralizing antibodies induces prostate cancer tumor regression in vivo. AB - The identification of novel tumor-interactive chemokines and the associated insights into the molecular and cellular basis of tumor-microenvironment interactions have continued to stimulate the development of targeted cancer therapeutics. Recently, we have identified monocyte chemoattractant protein 1 (MCP-1; CCL2) as a prominent regulator of prostate cancer growth and metastasis. Using neutralizing antibodies to human CCL2 (CNTO888) and the mouse homologue CCL2/JE (C1142), we show that treatment with anti-CCL2/JE antibody (2 mg/kg, twice weekly i.p.) attenuated PC-3Luc-mediated overall tumor burden in our in vivo model of prostate cancer metastasis by 96% at 5 weeks postintracardiac injection. Anti-CCL2 inhibition was not as effective as docetaxel (40 mg/kg, every week for 3 weeks) as a single agent, but inhibition of CCL2 in combination with docetaxel significantly reduced overall tumor burden compared with docetaxel alone, and induced tumor regression relative to initial tumor burden. These data suggest an interaction between tumor-derived chemokines and host-derived chemokines acting in cooperation to promote tumor cell survival, proliferation, and metastasis. PMID- 17909052 TI - c-Fos as a proapoptotic agent in TRAIL-induced apoptosis in prostate cancer cells. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)/Apo-2L promotes apoptosis in cancer cells while sparing normal cells. Although many cancers are sensitive to TRAIL-induced apoptosis, some evade the proapoptotic effects of TRAIL. Therefore, differentiating molecular mechanisms that distinguish between TRAIL-sensitive and TRAIL-resistant tumors are essential for effective cancer therapies. Here, we show that c-Fos functions as a proapoptotic agent by repressing the antiapoptotic molecule c-FLIP(L). c-Fos binds the c-FLIP(L) promoter, represses its transcriptional activity, and reduces c-FLIP(L) mRNA and protein levels. Therefore, c-Fos is a key regulator of c-FLIP(L), and activation of c-Fos determines whether a cancer cell will undergo cell death after TRAIL treatment. Strategies to activate c-Fos or inhibit c-FLIP(L) may potentiate TRAIL based proapoptotic therapies. PMID- 17909054 TI - Cell cycle dependent and schedule-dependent antitumor effects of sorafenib combined with radiation. AB - The antineoplastic drug sorafenib (BAY 43-9006) is a multikinase inhibitor that targets the serine-threonine kinase B-Raf as well as several tyrosine kinases. Given the numerous molecular targets of sorafenib, there are several potential anticancer mechanisms of action, including induction of apoptosis, cytostasis, and antiangiogenesis. We observed that sorafenib has broad activity in viability assays in several human tumor cell lines but selectively induces apoptosis in only some lines. Sorafenib was found to decrease Mcl-1 levels in most cell lines tested, but this decrease did not correlate with apoptotic sensitivity. Sorafenib slows cell cycle progression and prevents irradiated cells from reaching and accumulating at G2-M. In synchronized cells, sorafenib causes a reversible G1 delay, which is associated with decreased levels of cyclin D1, Rb, and phosphorylation of Rb. Although sorafenib does not affect intrinsic radiosensitivity using in vitro colony formation assays, it significantly reduces colony size. In HCT116 xenograft tumor growth delay experiments in mice, sorafenib alters radiation response in a schedule-dependent manner. Radiation treatment followed sequentially by sorafenib was found to be associated with the greatest tumor growth delay. This study establishes a foundation for clinical testing of sequential fractionated radiation followed by sorafenib in gastrointestinal and other malignancies. PMID- 17909053 TI - Malignant progression and blockade of angiogenesis in a murine transgenic model of neuroblastoma. AB - Targeted expression of MYCN to the neural crest [under control of the rat tyrosine hydroxylase (TH) promoter] causes neuroblastoma in transgenic mice (TH MYCN) and is a well-established model for this disease. Because high levels of MYCN are associated with enhanced tumor angiogenesis and poor clinical outcome in neuroblastoma, we serially characterized malignant progression, angiogenesis, and sensitivity to angiogenic blockade in tumors from these animals. Tumor cells were proliferative, secreted high levels of the angiogenic ligand vascular endothelial growth factor (VEGF), and recruited a complex vasculature expressing the angiogenic markers VEGF-R2, alpha-SMA, and matrix metalloproteinases MMP-2 and MMP-9, all of which are also expressed in human disease. Treatment of established murine tumors with the angiogenesis inhibitor TNP-470 caused near-complete ablation, with reduced proliferation, enhanced apoptosis, and vasculature disruption. Because TNP-470 has been associated with neurotoxicity, we tested the recently described water-soluble HPMA copolymer-TNP-470 conjugate (caplostatin), which showed comparable efficacy and was well tolerated without weight loss or neurotoxicity as measured by rotarod testing. This study highlights the importance of angiogenesis inhibition in a spontaneous murine tumor with native tumor-microenvironment interactions, validates the use of mice transgenic for TH MYCN as a model for therapy in this common pediatric tumor, and supports further clinical development of caplostatin as an antiangiogenic therapy in childhood neuroblastoma. PMID- 17909055 TI - Tumor necrosis factor alpha mediates homogeneous distribution of liposomes in murine melanoma that contributes to a better tumor response. AB - Successful treatment of solid tumors with chemotherapeutics requires that adequate levels reach the tumor cells. Tumor vascular normalization has been proposed to enhance drug delivery and improve tumor response to chemotherapy. Differently, augmenting leakage of the tumor-associated vasculature, and as such enhance vascular abnormality, may improve tumor response as well. In the present study, we show that addition of low-dose tumor necrosis factor alpha (TNF) to systemic injections with pegylated long circulating liposomes augmented the tumor accumulation of these liposomes 5- to 6-fold, which strongly correlated with enhanced tumor response. Using intravital microscopy, we could study the liposomal distribution inside the tumor in more detail. Especially 100 nm liposomes effectively extravasate in the surrounding tumor tissue in the presence of TNF and this occurred without any effect on tumor vascular density, branching, and diameter. Next to that, we observed in living animals that tumor cells take up the liposomes intact, followed by intracellular degradation. To our knowledge, this is an unprecedented observation. Taken together, TNF renders more tumor vessels permeable, leading to a more homogeneous distribution of the liposomes throughout the tumor, which is crucial for an optimal tumor response. We conclude that delivery of nanoparticulate drug formulations to solid tumor benefits from augmenting the vascular leakage through vascular manipulation with vasoactive drugs like TNF. PMID- 17909056 TI - Autocrine glutamate signaling promotes glioma cell invasion. AB - Malignant gliomas have been shown to release glutamate, which kills surrounding brain cells, creating room for tumor expansion. This glutamate release occurs primarily via system xC, a Na+-independent cystine-glutamate exchanger. We show here, in addition, that the released glutamate acts as an essential autocrine/paracrine signal that promotes cell invasion. Specifically, chemotactic invasion and scrape motility assays each show dose-dependent inhibition of cell migration when glutamate release was inhibited using either S-(4)-CPG or sulfasalazine, both potent blockers of system xC. This inhibition could be overcome by the addition of exogenous glutamate (100 micromol/L) in the continued presence of the inhibitors. Migration/invasion was also inhibited when Ca2+ permeable alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptors (AMPA-R) were blocked using GYKI or Joro spider toxin, whereas CNQX was ineffective. Ca2+ imaging experiments show that the released glutamate activates Ca2+-permeable AMPA-R and induces intracellular Ca2+ oscillations that are essential for cell migration. Importantly, glioma cells release glutamate in sufficient quantities to activate AMPA-Rs on themselves or neighboring cells, thus acting in an autocrine and/or paracrine fashion. System xC and the appropriate AMPA-R subunits are expressed in all glioma cell lines, patient derived glioma cells, and acute patient biopsies investigated. Furthermore, animal studies in which human gliomas were xenographed into scid mice show that chronic inhibition of system xC-mediated glutamate release leads to smaller and less invasive tumors compared with saline-treated controls. These data suggest that glioma invasion is effectively disrupted by inhibiting an autocrine glutamate signaling loop with a clinically approved candidate drug, sulfasalazine, already in hand. PMID- 17909057 TI - Inhibitors of ubiquitin-activating enzyme (E1), a new class of potential cancer therapeutics. AB - The conjugation of proteins with ubiquitin plays numerous regulatory roles through both proteasomal-dependent and nonproteasomal-dependent functions. Alterations in ubiquitylation are observed in a wide range of pathologic conditions, including numerous malignancies. For this reason, there is great interest in targeting the ubiquitin-proteasome system in cancer. Several classes of proteasome inhibitors, which block degradation of ubiquitylated proteins, are widely used in research, and one, Bortezomib, is now in clinical use. Despite the well-defined and central role of the ubiquitin-activating enzyme (E1), no cell permeable inhibitors of E1 have been identified. Such inhibitors should, in principle, block all functions of ubiquitylation. We now report 4[4-(5-nitro furan-2-ylmethylene)-3,5-dioxo-pyrazolidin-1-yl]-benzoic acid ethyl ester (PYR 41) as the first such inhibitor. Unexpectedly, in addition to blocking ubiquitylation, PYR-41 increased total sumoylation in cells. The molecular basis for this is unknown; however, increased sumoylation was also observed in cells harboring temperature-sensitive E1. Functionally, PYR-41 attenuates cytokine mediated nuclear factor-kappaB activation. This correlates with inhibition of nonproteasomal (Lys-63) ubiquitylation of TRAF6, which is essential to IkappaB kinase activation. PYR-41 also prevents the downstream ubiquitylation and proteasomal degradation of IkappaBalpha. Furthermore, PYR-41 inhibits degradation of p53 and activates the transcriptional activity of this tumor suppressor. Consistent with this, it differentially kills transformed p53-expressing cells. Thus, PYR-41 and related pyrazones provide proof of principle for the capacity to differentially kill transformed cells, suggesting the potential for E1 inhibitors as therapeutics in cancer. These inhibitors can also be valuable tools for studying ubiquitylation. PMID- 17909058 TI - Heat shock protein 90alpha recruits FLIPS to the death-inducing signaling complex and contributes to TRAIL resistance in human glioma. AB - Heat shock protein 90 (HSP90) is a molecular chaperone that contributes to the proper folding and stability of target proteins. Because HSP90 has been suggested to interact with FLIP(S), the key regulator of tumor necrosis factor-alpha related apoptosis-inducing ligand (TRAIL)-induced apoptosis in glioma cells, we examined the role HSP90 played in controlling TRAIL response. HSP90alpha was found to associate with FLIP(S) in resting cells in a manner dependent on the ATP binding NH2-terminal domain of HSP90alpha. Following TRAIL exposure, HSP90alpha and the client FLIP(S) protein were recruited to the death-inducing signaling complex (DISC). Short interfering RNA-mediated suppression of HSP90alpha did not alter the total cellular levels of FLIP(S), but rather inhibited the recruitment of FLIP(S) and other antiapoptotic proteins such as RIP and FLIP(L) to the DISC, and sensitized otherwise resistant glioma cells to TRAIL-induced apoptosis. These results show that HSP90alpha, by localizing FLIP(S) to the DISC, plays a key role in the resistance of tumor cells to TRAIL, and perhaps other proapoptotic agents. The results also define a novel means of apoptotic control by a HSP90alpha that may in turn help explain the global antiapoptotic effects of this protein. PMID- 17909059 TI - The multikinase inhibitor sorafenib potentiates TRAIL lethality in human leukemia cells in association with Mcl-1 and cFLIPL down-regulation. AB - Interactions between the multikinase inhibitor sorafenib and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) were examined in malignant hematopoietic cells. Pretreatment (24 h) of U937 leukemia cells with 7.5 micromol/L sorafenib dramatically increased apoptosis induced by sublethal concentrations of TRAIL/Apo2L (75 ng/mL). Similar interactions were observed in Raji, Jurkat, Karpas, K562, U266 cells, primary acute myelogenous leukemia blasts, but not in normal CD34+ bone marrow cells. Sorafenib/TRAIL-induced cell death was accompanied by mitochondrial injury and release of cytochrome c, Smac, and AIF into the cytosol and caspase-9, caspase-3, caspase-7, and caspase-8 activation. Sorafenib pretreatment down-regulated Bcl-xL and abrogated Mcl-1 expression, whereas addition of TRAIL sharply increased Bid activation, conformational change of Bak (ccBak) and Bax (ccBax), and Bax translocation. Ectopic Mcl-1 expression significantly attenuated sorafenib/TRAIL-mediated lethality and dramatically reduced ccBak while minimally affecting levels of ccBax. Similarly, inhibition of the receptor-mediated apoptotic cascade with a caspase-8 dominant-negative mutant significantly blocked sorafenib/TRAIL-induced lethality but not Mcl-1 down-regulation or Bak/Bax conformational change, indicating that TRAIL-mediated receptor pathway activation is required for maximal lethality. Sorafenib/TRAIL did not increase expression of DR4/DR5, or recruitment of procaspase-8 or FADD to the death-inducing signaling complex (DISC), but strikingly increased DISC-associated procaspase-8 activation. Sorafenib also down-regulated cFLIP(L), most likely through a translational mechanism, in association with diminished eIF4E phosphorylation, whereas ectopic expression of cFLIP(L) significantly reduced sorafenib/TRAIL lethality. Together, these results suggest that in human leukemia cells, sorafenib potentiates TRAIL induced lethality by down-regulating Mcl-1 and cFLIP(L), events that cooperate to engage the intrinsic and extrinsic apoptotic cascades, culminating in pronounced mitochondrial injury and apoptosis. PMID- 17909060 TI - Local administration of interleukin-11 ameliorates intestinal radiation injury in rats. AB - Intestinal radiation injury is dose limiting during abdominal and pelvic radiotherapy and critical for the outcome after accidental whole-body radiation exposure. The multifunctional cytokine, interleukin-11 (IL-11), ameliorates the intestinal radiation response, but its clinical use is hampered by severe toxicity after systemic administration. This study addressed whether protection against intestinal radiation injury can be achieved by intraluminal administration of IL-11. Male rats underwent surgical transposition of a 4-cm small bowel loop to the scrotum. For repeated intraluminal drug administration, an ileostomy, proximal to the bowel loop in the scrotum, was created. The transposed intestinal loop was exposed to 5 Gy fractions on 9 consecutive days. Recombinant human IL-11 (rhIL-11; 2 mg/kg/d) or vehicle was given through the ileostomy from 2 days before until 2 weeks after irradiation. At 2 weeks, structural, cellular, and molecular aspects of intestinal radiation injury were assessed. rhIL-11 ameliorated structural manifestations of radiation enteropathy, including radiation injury score (6.5 +/- 0.6 in the vehicle group versus 4.0 +/- 0.3 in the IL-11 group; P = 0.001), mucosal surface area loss (0.2 +/- 0.1 versus 0.5 +/- 0.03; P < 0.0001), and intestinal wall thickening (842 +/- 66 microm versus 643 +/- 54 microm; P = 0.02), reduced postradiation transforming growth factor-beta overexpression, and reduced numbers of ED2-positive cells. Postirradiation mucosal mast cell numbers were partially restored by rhIL-11. These data show that local administration of rhIL-11 ameliorates early intestinal radiation injury and support further development of rhIL-11 to reduce manifestations of intestinal radiation injury in the clinic. PMID- 17909061 TI - Phage display derived human monoclonal antibodies isolated by binding to the surface of live primary breast cancer cells recognize GRP78. AB - Clinical trials using monoclonal antibodies (mAb) against cell-surface markers have yielded encouraging therapeutic results in several cancer types. Generally, however, anticancer antibodies are only efficient against a subpopulation of cancers, and there is a strong need for identification of novel targets and human antibodies against them. We have isolated single-chain human mAbs from a large naive antibody phage display library by panning on a single-cell suspension of freshly isolated live cancer cells from a human breast cancer specimen, and these antibodies were shown to specifically recognize cancer-associated cell-surface proteins. One of the isolated human antibody fragments, Ab39, recognizes a cell surface antigen expressed on a subpopulation of cancer cell lines of different origins. Immunohistochemical analysis of a large panel of cancerous and normal tissues showed that Ab39 bound strongly to several cancers, including 45% breast carcinomas, 35% lung cancers, and 86% melanomas, but showed no or weak binding to normal tissues. A yeast two-hybrid screen of a large human testis cDNA library identified the glucose-regulated protein of 78 kDa (GRP78) as the antigen recognized by Ab39. The interaction was confirmed by colocalization studies and antibody competition experiments that also mapped the epitope recognized by Ab39 to the COOH terminus of GRP78. The expression of GRP78 on the surface of cancer cells, but not normal cells, makes it an attractive target for cancer therapies including mAb-based immunotherapy. Our results suggest that the human antibody Ab39 may be a useful starting point for further genetic optimization that could render it a useful diagnostic and therapeutic reagent for a variety of cancers. PMID- 17909062 TI - Dynamics of the immune reaction to pancreatic cancer from inception to invasion. AB - The dynamics of cancer immunosurveillance remain incompletely understood, hampering efforts to develop immunotherapy of cancer. We evaluated the evolving in vivo immune response to a spontaneous tumor in a genetically defined mouse model of pancreatic ductal adenocarcinoma from the inception of preinvasive disease to invasive cancer. We observed a prominent leukocytic infiltration even around the lowest grade preinvasive lesions, but immunosuppressive cells, including tumor-associated macrophages, myeloid-derived suppressor cells (MDSC), and regulatory T cells (Treg), dominated the early response and persisted through invasive cancer. Effector T cells, however, were scarce in preinvasive lesions, found in only a subset of advanced cancers, and showed no evidence of activation. The lack of tumor-infiltrating effector T cells strongly correlated with the presence of intratumoral MDSC with a near mutual exclusion. In vitro, we found that MDSC suppressed T-cell proliferation. Overall, our results show that suppressive cells of the host immune system appear early during pancreatic tumorigenesis, preceding and outweighing antitumor cellular immunity, and likely contribute to disease progression. Thus, in contrast to the hypothesis that an early "elimination phase" of cancer immunosurveillance is eventually overwhelmed by a growing invasive tumor, our findings suggest that productive tumor immunity may be undermined from the start. Efforts to test potent inhibitors of MDSC, tumor-associated macrophages, and Treg, particularly early in the disease represent important next steps for developing novel immunotherapy of cancer. PMID- 17909063 TI - A placenta-specific gene ectopically activated in many human cancers is essentially involved in malignant cell processes. AB - The identification and functional characterization of tumor-specific genes is a prerequisite for the development of targeted cancer therapies. Using an integrated data mining and experimental validation approach for the discovery of new targets for antibody therapy of cancer, we identified PLAC1. PLAC1 is a placenta-specific gene with no detectable expression in any other normal human tissue. However, it is frequently aberrantly activated and highly expressed in a variety of tumor types, in particular breast cancer. RNAi-mediated silencing of PLAC1 in MCF-7 and BT-549 breast cancer cells profoundly impairs motility, migration, and invasion and induces a G1-S cell cycle block with nearly complete abrogation of proliferation. Knockdown of PLAC1 is associated with decreased expression of cyclin D1 and reduced phosphorylation of AKT kinase. Moreover, PLAC1 is localized on the surface of cancer cells and is accessible for antibodies which antagonize biological functions of this molecule. These features, in summary, make PLAC1 an attractive candidate for targeted immunotherapeutic approaches. PMID- 17909064 TI - Enhancement of antibody-dependent mechanisms of tumor cell lysis by a targeted activator of complement. AB - Complement inhibitors expressed on tumor cells provide a hindrance to the therapeutic efficacy of some monoclonal antibodies (mAb). We investigated a novel strategy to overwhelm complement inhibitor activity and amplify complement activation on tumor cells. The C3-binding domain of human complement receptor 2 (CR2; CD21) was linked to the complement-activating Fc region of human IgG1 (CR2 Fc), and the ability of the construct to target and amplify complement deposition on tumor cells was investigated. CR2 binds C3 activation fragments, and CR2-Fc targeted tumor cells by binding to C3 initially deposited by a tumor-specific antibody. Complement deposition on Du145 cells (human prostate cancer cell line) and anti-MUC1 mAb-mediated complement-dependent lysis of Du145 cells were significantly enhanced by CR2-Fc. Anti-MUC1 antibody-dependent cell-mediated cytotoxicity of Du145 by human peripheral blood mononuclear cells was also significantly enhanced by CR2-Fc in both the presence and the absence of complement. Radiolabeled CR2-Fc targeted to s.c. Du145 tumors in nude mice treated with anti-MUC1 mAb, validating the targeting strategy in vivo. A metastatic model was used to investigate the effect of CR2-Fc in a therapeutic paradigm. Administration of CR2-Fc together with mAb therapy significantly improved long-term survival of nude mice challenged with an i.v. injection of EL4 cells. The data show that CR2-Fc enhances the therapeutic efficacy of antibody therapy, and the construct may provide particular benefits under conditions of limiting antibody concentration or low tumor antigen density. PMID- 17909065 TI - Accelerated bone resorption, due to dietary calcium deficiency, promotes breast cancer tumor growth in bone. AB - The skeleton is a major site of breast cancer metastases. High bone turnover increases risk of disease progression and death. However, there is no direct evidence that high bone turnover is causally associated with the establishment and progression of metastases. In this study, we investigate the effects of high bone turnover in a model of breast cancer growth in bone. Female nude mice commenced a diet containing normal (0.6%; 'Normal-Ca') or low (0.1%; 'Low-Ca') calcium content. Mice were concurrently treated with vehicle or osteoprotegerin (1 mg/kg/d s.c; n = 16 per group). Three days later (day 0), 50,000 Tx-SA cells (variant of MDA-MB-231 cells) were implanted by intratibial injection. On day 0, mice receiving Low-Ca had increased serum parathyroid hormone (PTH) and tartrate resistant acid phosphatase 5b levels, indicating secondary hyperparathyroidism and high bone turnover, which was maintained until day 17. Osteoprotegerin increased serum PTH but profoundly reduced bone resorption. On day 17, in mice receiving Low-Ca alone, lytic lesion area, tumor area, and cancer cell proliferation increased by 43%, 24%, and 24%, respectively, compared with mice receiving Normal Ca (P < 0.01). Osteoprotegerin treatment completely inhibited lytic lesions, reduced tumor area, decreased cancer cell proliferation, and increased cancer cell apoptosis. Increased bone turnover, due to dietary calcium deficiency, promotes tumor growth in bone, independent of the action of PTH. Breast cancer patients frequently have low dietary calcium intake and high bone turnover. Treatment to correct calcium insufficiency and/or treatment with antiresorptive agents, such as osteoprotegerin, may be of benefit in the adjuvant as well as palliative setting. PMID- 17909066 TI - Definition of functionally important mechanistic differences among selective estrogen receptor down-regulators. AB - One subclass of antiestrogens, the selective estrogen receptor down-regulators (SERDs), have received considerable attention of late as they competitively inhibit estrogen binding and induce a rapid, proteasome-dependent degradation of the receptor. Contained within this class of molecules is the steroidal antiestrogen ICI182,780 (faslodex), recently approved for the treatment of metastatic cancer, and GW5638/DPC974, a SERD that is currently being evaluated in the clinic. Given that mechanistic differences between different selective estrogen receptor modulators have been translated into important clinical profiles, it was of interest to determine if the SERD subclass of ligands were likewise functionally or mechanistically distinguishable. In this study, we show that although the steroidal and nonsteroidal SERDs target ERalpha for degradation, the underlying mechanism(s) are different. Of note was the identification of a specific protein-protein interaction surface presented on ERalpha in the presence of the ICI182,780-activated receptor which is required for degradation. Interestingly, this surface is also presented on ERalpha in the presence of RU58,668, a SERD that is chemically distinct from ICI182,780. This surface is not required for GW5638-mediated degradation, and thus, this SERD seems to affect ERalpha down-regulation by a different mechanism. These data suggest that sequencing of therapies using drugs of this class is likely to be possible. Finally, because of the unmet need for orally active SERDS that function similarly to ICI182,780, we have used the insights from these mechanistic studies to develop and validate a high-throughput screen for compounds of this class with improved pharmaceutical properties. PMID- 17909067 TI - Association study of 69 genes in the ret pathway identifies low-penetrance loci in sporadic medullary thyroid carcinoma. AB - To date, few association studies have been done to better understand the genetic basis for the development of sporadic medullary thyroid carcinoma (sMTC). To identify additional low-penetrance genes, we have done a two-stage case-control study in two European populations using high-throughput genotyping. We selected 417 single nucleotide polymorphisms (SNP) belonging to 69 genes either related to RET signaling pathway/functions or involved in key processes for cancer development. TagSNPs and functional variants were included where possible. These SNPs were initially studied in the largest known series of sMTC cases (n = 266) and controls (n = 422), all of Spanish origin. In stage II, an independent British series of 155 sMTC patients and 531 controls was included to validate the previous results. Associations were assessed by an exhaustive analysis of individual SNPs but also considering gene- and linkage disequilibrium-based haplotypes. This strategy allowed us to identify seven low-penetrance genes, six of them (STAT1, AURKA, BCL2, CDKN2B, CDK6, and COMT) consistently associated with sMTC risk in the two case-control series and a seventh (HRAS) with individual SNPs and haplotypes associated with sMTC in the Spanish data set. The potential role of CDKN2B was confirmed by a functional assay showing a role of a SNP (rs7044859) in the promoter region in altering the binding of the transcription factor HNF1. These results highlight the utility of association studies using homogeneous series of cases for better understanding complex diseases. PMID- 17909068 TI - SND1, a component of RNA-induced silencing complex, is up-regulated in human colon cancers and implicated in early stage colon carcinogenesis. AB - Colon cancers have been shown to develop after accumulation of multiple genetic and epigenetic alterations with changes in global gene expression profiles, contributing to the establishment of widely diverse phenotypes. Transcriptional and posttranscriptional regulation of gene expression by small RNA species, such as the small interfering RNA and microRNA and the RNA-induced silencing complex (RISC), is currently drawing major interest with regard to cancer development. SND1, also called Tudor-SN and p100 and recently reported to be a component of RISC, is among the list of highly expressed genes in human colon cancers. In the present study, we showed remarkable up-regulation of SND1 mRNA in human colon cancer tissues, even in early-stage lesions, and also in colon cancer cell lines. When mouse Snd1 was stably overexpressed in IEC6 rat intestinal epithelial cells, contact inhibition was lost and cell growth was promoted, even after the cells became confluent. Intriguingly, IEC6 cells with high levels of Snd1 also showed an altered distribution of E-cadherin from the cell membrane to the cytoplasm, suggesting loss of cellular polarity. Furthermore, the adenomatous polyposis coli (Apc) protein was coincidentally down-regulated, with no significant changes in the Apc mRNA level. Immunohistochemical analysis using chemically induced colonic lesions developed in rats revealed overexpression of Snd1 not only in colon cancers but also in aberrant crypt foci, putative precancerous lesions of the colon. Up-regulation of SND1 may thus occur at a very early stage in colon carcinogenesis and contribute to the posttranscriptional regulation of key players in colon cancer development, including APC and beta-catenin. PMID- 17909069 TI - Smad7 sensitizes tumor necrosis factor induced apoptosis through the inhibition of antiapoptotic gene expression by suppressing activation of the nuclear factor kappaB pathway. AB - Although tumor necrosis factor (TNF) induces apoptosis and cell death in many tumor cells, some cancer cells are still resistant to the TNF-induced death signal. In this report, we showed that Smad7, an inhibitory Smad of transforming growth factor-beta (TGF-beta) signaling, can overcome the TNF resistance in human breast and gastric cancer cells. Overexpression of Smad7 induces the degradation of poly(ADP-ribose) polymerase and the activation of caspase cascade. Although c Jun NH2-terminal kinase (JNK) signaling is involved in TNF-induced cell death, the expression of Smad7 does not synergize the activation of JNK. However, the activation of nuclear factor-kappaB (NF-kappaB), the cell survival factor, is markedly decreased in Smad7-stable cells. Furthermore, the expression of antiapoptotic target genes of NF-kappaB is significantly reduced in accordance with the level of Smad7. In addition, Smad7 mediates the inhibitory activity of TGF-beta on TNF-induced NF-kappaB activation and the synergistic activity of TGF beta on TNF-induced apoptosis. These findings suggest that Smad7 sensitizes the tumor cells to TNF-induced apoptosis through the inhibition of expression of antiapoptotic NF-kappaB target genes. PMID- 17909070 TI - Do MDM2 SNP309 and TP53 R72P interact in breast cancer susceptibility? A large pooled series from the breast cancer association consortium. AB - Association studies in large series of breast cancer patients can be used to identify single-nucleotide polymorphisms (SNP) contributing to breast cancer susceptibility. Previous studies have suggested associations between variants in TP53 (R72P) and MDM2 (SNP309) and cancer risk. Data from molecular studies suggest a functional interaction between these genes. We therefore investigated the effect of TP53 R72P and MDM2 SNP309 on breast cancer risk and age at onset of breast cancer in a pooled series of 5,191 cases and 3,834 controls from the Breast Cancer Association Consortium (BCAC). Breast cancer risk was not found to be associated with the combined variant alleles [odds ratio (OR), 1.00; 95% confidence interval (95% CI), 0.81-1.23]. Estimated ORs were 1.01 (95% CI, 0.93 1.09) per MDM2 SNP309 allele and 0.98 (95% CI, 0.91-1.04) for TP53 R72P. Although we did find evidence for a 4-year earlier age at onset for carriers of both variant alleles in one of the breast cancer patient series of the BCAC (the German series), we were not able to confirm this effect in the pooled analysis. Even so, carriers of both variant alleles did not have different risk estimates for bilateral or estrogen receptor-positive breast cancer. In conclusion, in this large collaborative study, we did not find an association of MDM2 SNP309 and TP53 R72P, separately or in interaction, with breast cancer. This suggests that any effect of these two variants would be very small and possibly confined to subgroups that were not assessed in our present study. PMID- 17909071 TI - Genetic heterogeneity among Fanconi anemia heterozygotes and risk of cancer. AB - Fanconi anemia (FA) is a rare autosomal recessive disease characterized by a greatly increased risk of cancer among those diagnosed with the syndrome. The question as to whether FA heterozygotes are at increased risk for cancer is of great importance to those at risk for being a carrier. To address this question, we formed a cohort of grandparents of probands identified through the International Fanconi Anemia Registry. We obtained informed consent, a short questionnaire, and either blood or buccal swab DNA. After diagnosis of the proband was confirmed and complementation studies or DNA sequencing on the proband were completed, mutation analyses of the putative carriers and noncarriers was carried out. Standardized incidence ratios (SIR) were calculated to compare the observed cancer incidence of the grandparents and other relatives with the expected rates of cancer, using the Surveillance, Epidemiology, and End Results registries and the Connecticut Cancer registry. In the 944 study subjects who participated (784 grandparents and 160 other relatives), there was no suggestion of an increase in overall cancer incidence. On the other hand, a significantly higher rate of breast cancer than expected was observed among carrier grandmothers [SIR, 1.7; 95% confidence interval (95% CI), 1.1-2.7]. Among the grandmothers, those who were carriers of FANCC mutations were found to be at highest risk (SIR, 2.4; 95% CI, 1.1-5.2). Overall, there was no increased risk for cancer among FA heterozygotes in this study of Fanconi relatives, although there is some evidence that FANCC mutations are possibly breast cancer susceptibility alleles. PMID- 17909072 TI - The cooked meat derived genotoxic carcinogen 2-amino-3-methylimidazo[4,5 b]pyridine has potent hormone-like activity: mechanistic support for a role in breast cancer. AB - The cooked meat-derived heterocyclic amine 2-amino-3-methylimidazo[4,5-b]pyridine (PhIP) is activated by CYP1A2 to the N-hydroxy metabolite, then esterified by acetyl transferase and sulfur transferase into unstable DNA-reactive products that can lead to mutation. The genotoxicity of PhIP has been implicated in its carcinogenicity. Yet, CYP1A2-null mice are still prone to PhIP-mediated cancer, inferring that alternative mechanisms must be operative in tumor induction. PhIP induces tumors of the breast, prostate, and colon in rats and lymphoma in mice. This profile of carcinogenicity is indicative of hormonal involvement. We recently reported that PhIP has potent estrogenic activity inducing transcription of estrogen (E2)-regulated genes, proliferation of E(2)-dependent cells, up regulation of progesterone receptor, and stimulation of mitogen-activated protein kinase signaling. In this report, we show for the first time that PhIP at doses as low as of 10(-11) mol/L has direct effects on a rat pituitary lactotroph model (GH3 cells) and is able to induce cell proliferation and the synthesis and secretion of prolactin. This PhIP-induced pituitary cell proliferation and synthesis and secretion of prolactin can be attenuated by an estrogen receptor (ER) inhibitor, implying that PhIP effects on lactotroph responses are ERalpha mediated. In view of the strong association between estrogen, progesterone, prolactin, and breast cancer, the PhIP repertoire of hormone-like activities provides further mechanistic support for the tissue-specific carcinogenicity of the chemical. Furthermore, the recent epidemiology studies that report an association between consumption of cooked red meat and premenopausal and postmenopausal human breast cancer are consonant with these observations. PMID- 17909073 TI - Comment on: Screening for Lynch Syndrome (Hereditary Nonpolyposis Colorectal Cancer) among Endometrial Cancer Patients. PMID- 17909075 TI - Macrophages contribute to the antitumor activity of the anti-CD30 antibody SGN 30. AB - Increased expression of CD30 is associated with a variety of hematologic malignancies, including Hodgkin disease (HD) and anaplastic large cell lymphoma (ALCL). The anti-CD30 monoclonal antibody SGN-30 induces direct antitumor activity by promoting growth arrest and DNA fragmentation of CD30(+) tumor cells. In this study, we investigated the contributions of Fc-mediated effector cell functions to SGN-30 activity. We determined that antibody-dependent cellular phagocytosis, mediated by macrophages, to contribute significantly to antitumor activity in vitro. To delineate the identity of the host effector cells involved in mediating antitumor activity in vivo, we studied the effects of effector cell ablation in a disseminated model of HD (L540cy). Depletion of macrophages markedly reduced efficacy of SGN-30, demonstrating that macrophages contribute significantly to SGN-30 efficacy in this model. These findings may have implications for patient stratification or combination treatment strategies in clinical trials conducted with SGN-30 in HD and ALCL. PMID- 17909074 TI - Postsurgery outcomes in patients with polycythemia vera and essential thrombocythemia: a retrospective survey. AB - A multicenter retrospective analysis was performed to estimate the frequency of thrombosis and hemorrhage after surgical procedures in patients with polycythemia vera (PV) and patients with essential thrombocythemia (ET). Data from 105 patients with PV and 150 patients with ET were analyzed, for a total of 311 surgical interventions. An emergency procedure was performed in 25 (8.1%) patients; 194 surgeries were done under general anesthesia, and 21 (23%) of 91 abdominal interventions were done under laparoscopy; 155 (50.1%) were major surgeries. Subcutaneous heparin was administered in 169 (54.3%) of 311 cases and antiplatelet therapy in 48 (15.4%) of 311 case interventions. One hundred eighty eight (74%) of 255 patients were on cytoreductive therapy before surgery. No events were observed in 259 (83.2%) of 311 procedures during 3 months of follow up; there were 12 arterial and 12 venous thrombotic events, 23 major and 7 minor hemorrhages, and 5 deaths. Arterial thromboses were more frequent in ET (5.3% vs 1.5%; P=.08), venous events were more frequent in PV (7.7% vs 1.1%; P=.002). There was not a correlation between bleeding episodes and the type of diagnosis, use of antithrombotic prophylaxis, or type of surgery. A high proportion of PV and ET surgeries was complicated by vascular occlusion (7.7%) or by a major hemorrhage (7.3%). Prospective investigations analyzing the optimal prophylaxis in these patients are suggested. PMID- 17909076 TI - Prediction of adverse outcomes in children with sickle cell anemia: a study of the Dallas Newborn Cohort. AB - The Cooperative Study of Sickle Cell Disease reported that dactylitis, severe anemia, and leukocytosis in very young children with sickle cell disease (SCD) increased the risk of later adverse outcomes, including death, stroke, frequent pain, and recurrent acute chest syndrome. This model has not been validated in other cohorts. We evaluated its performance in the Dallas Newborn Cohort, a newborn inception cohort of children with SCD. We studied 168 children (55% male, 97% sickle cell anemia) with a mean follow-up of 7.1 years who provided 1188 patient-years of observation. Of the 23 (13.7%) subjects who experienced adverse events, 2 (1.2%) died, 14 (8.3%) had a stroke, 4 (2.4%) had frequent pain, and 3 (1.8%) had recurrent acute chest syndrome. No relationship existed between early clinical predictors and later adverse outcomes, with the possible exception of leukocyte count. Most subjects who experienced adverse events were predicted to be at low risk for those events. No subject who was predicted to be at high risk actually experienced an adverse outcome. The sensitivity of the model did not rise above 20% until specificity fell below 60%. We suggest that this model not be used as a criterion to initiate early interventions for SCD. PMID- 17909077 TI - Early peripheral blood blast clearance during induction chemotherapy for acute myeloid leukemia predicts superior relapse-free survival. AB - In childhood acute lymphoblastic leukemia (ALL), a rapid decline of circulating leukemic blasts in response to induction chemotherapy or prednisone is one of the most important prognostic factors, not only for achieving remission but also for relapse-free survival (RFS). However, in acute myeloid leukemia (AML) parameters of chemosensitivity have been restricted mainly to the rapidity of achievement of complete remission (CR) or the assessment of residual leukemic bone marrow blasts during aplasia. We hypothesized that the time to circulating peripheral blood blast clearance, as a potential surrogate for in vivo chemosensitivity, would have prognostic relevance in AML also. In a retrospective analysis of a cohort of 86 adult patients with AML receiving uniform induction and consolidation chemotherapy, we demonstrate that the time to clearance of circulating blasts during induction chemotherapy is an independent prognostic marker of RFS, superseding other known or established risk factors, including karyotype and number of inductions to achieve CR. PMID- 17909078 TI - Calmodulin-dependent kinase IV links Toll-like receptor 4 signaling with survival pathway of activated dendritic cells. AB - Microbial products, including lipopolysaccharide (LPS), an agonist of Toll-like receptor 4 (TLR4), regulate the lifespan of dendritic cells (DCs) by largely undefined mechanisms. Here, we identify a role for calcium-calmodulin-dependent kinase IV (CaMKIV) in this survival program. The pharmacologic inhibition of CaMKs as well as ectopic expression of kinase-inactive CaMKIV decrease the viability of monocyte-derived DCs exposed to bacterial LPS. The defect in TLR4 signaling includes a failure to accumulate the phosphorylated form of the cAMP response element-binding protein (pCREB), Bcl-2, and Bcl-xL. CaMKIV null mice have a decreased number of DCs in lymphoid tissues and fail to accumulate mature DCs in spleen on in vivo exposure to LPS. Although isolated Camk4-/- DCs are able to acquire the phenotype typical of mature cells and release normal amounts of cytokines in response to LPS, they fail to accumulate pCREB, Bcl-2, and Bcl-xL and therefore do not survive. The transgenic expression of Bcl-2 in CaMKIV null mice results in full recovery of DC survival in response to LPS. These results reveal a novel link between TLR4 and a calcium-dependent signaling cascade comprising CaMKIV-CREB-Bcl-2 that is essential for DC survival. PMID- 17909079 TI - HIV-1 induced activation of CD4+ T cells creates new targets for HIV-1 infection in human lymphoid tissue ex vivo. AB - We demonstrate mechanisms by which HIV-1 appears to facilitate its own infection in ex vivo-infected human lymphoid tissue. In this system, HIV-1 readily infects various CD4+ T cells, but productive viral infection was supported predominantly by activated T cells expressing either CD25 or HLA-DR or both (CD25/HLA-DR) but not other activation markers: There was a strong positive correlation (r=0.64, P=.001) between virus production and the number of CD25+/HLA-DR+ T cells. HIV-1 infection of lymphoid tissue was associated with activation of both HIV-1 infected and uninfected (bystanders) T cells. In these tissues, apoptosis was selectively increased in T cells expressing CD25/HLA-DR and p24gag but not in cells expressing either of these markers alone. In the course of HIV-1 infection, there was a significant increase in the number of activated (CD25+/HLA-DR+) T cells both infected and uninfected (bystander). By inducing T cells to express particular markers of activation that create new targets for infection, HIV-1 generates in ex vivo lymphoid tissues a vicious destructive circle of activation and infection. In vivo, such self-perpetuating cycle could contribute to HIV-1 disease. PMID- 17909081 TI - Immunogenicity of umbilical cord tissue derived cells. AB - Umbilical cord tissue provides a unique source of cells with potential for tissue repair. Umbilical cord tissue-derived cells (UTCs) are MHC class I (MHCI) dull and negative for MHC class II (MHCII), but can be activated to increase MHCI and to express MHCII with IFN-gamma stimulation. Mesenchymal stem cells with similar characteristics have been inferred to be nonimmunogenic; however, in most cases, immunogenicity was not directly assessed. Using UTC from Massachusetts General Hospital MHC-defined miniature swine, we assessed immunogenicity across a full MHC barrier. Immunogenicity was assessed by in vitro assays including mixed lymphocyte reaction (MLR) and flow cytometry to detect serum alloantibody. A single injection of MHC-mismatched unactivated UTCs did not induce a detectable immune response. When injected in an inflamed region, injected repeatedly in the same region or stimulated with IFN-gamma prior to injection, UTCs were immunogenic. As clinical cellular repair strategies may involve injection of allogeneic cells into inflamed regions of damaged tissue or repeated doses of cells to achieve the desired benefit, our results on the immunogenicity of these cells in these circumstances may have important implications for optimal success and functional improvement for this cellular treatment strategy for diseased tissues. PMID- 17909080 TI - A2A receptor signaling promotes peripheral tolerance by inducing T-cell anergy and the generation of adaptive regulatory T cells. AB - Tissue-derived adenosine, acting via the adenosine A(2A) receptor (A(2A)R), is emerging as an important negative regulator of T-cell function. In this report, we demonstrate that A(2A)R stimulation not only inhibits the generation of adaptive effector T cells but also promotes the induction of adaptive regulatory T cells. In vitro, antigen recognition in the setting of A(2A)R engagement induces T-cell anergy, even in the presence of costimulation. T cells initially stimulated in the presence of an A(2A)R agonist fail to proliferate and produce interleukin-2 and interferon (IFN)-gamma when rechallenged in the absence of A(2A)R stimulation. Likewise, in an in vivo model of autoimmunity, tissue-derived adenosine promotes anergy and abrogates tissue destruction. Indeed, A(2A)R stimulation inhibits interleukin-6 expression while enhancing the production of transforming growth factor-beta. Accordingly, treating mice with A(2A)R agonists not only inhibits Th1 and Th17 effector cell generation but also promotes the generation of Foxp3(+) and LAG-3(+) regulatory T cells. In this regard, A(2A)R agonists fail to prevent autoimmunity by LAG-3(-/-) clonotypic T cells, implicating an important role for LAG-3 in adenosine-mediated peripheral tolerance. Overall, our findings demonstrate that extracellular adenosine stimulates the A(2A)R to promote long-term T-cell anergy and the generation of adaptive regulatory T cells. PMID- 17909082 TI - Antimetastatic effect of a small-molecule vacuolar H+-ATPase inhibitor in in vitro and in vivo preclinical studies. AB - On the basis of the evidence that vacuolar H(+)-ATPase is implicated in the development of the metastatic phenotype, we have explored the possibility to target the enzyme function in an attempt to control the metastatic behavior of tumor cells. In this study, we used an indole derivative, NiK-12192 [4-(5,6 dichloro-1H-indol-2-yl)-3-ethoxy-N-(2,2,6,6-tetramethyl-piperidin-4-yl) benzamide], recently identified as an effective inhibitor of vacuolar H(+) ATPase, as a potential antimetastatic agent in the treatment of NSCLC H460 xenograft, which is able to induce lung metastases in mice. Oral administration of NiK-12192 caused a significant inhibition of formation of spontaneous metastases. In contrast, the drug exhibited a negligible effect on the development of artificial metastases (i.e., after i.v. injection of tumor cells), thus supporting that the drug affects the early events of the metastatic process (e.g., migration and invasion). Cellular effects are consistent with this interpretation. In conclusion, the available results show for the first time that a vacuolar H(+)-ATPase inhibitor is effective in modulation of the metastatic behavior of a lung carcinoma, supporting its potential therapeutic interest as a novel treatment approach. PMID- 17909083 TI - Effect of puberty on the pharmacodynamic and pharmacokinetic properties of insulin pump therapy in youth with type 1 diabetes. PMID- 17909084 TI - Addition of pioglitazone and ramipril to intensive insulin therapy in type 2 diabetic patients improves vascular dysfunction by different mechanisms. AB - OBJECTIVE: We examined the relationship between glycemic control, vascular reactivity, and inflammation in type 2 diabetic subjects. RESEARCH DESIGN AND METHODS: Thirty subjects with type 2 diabetes were initiated on intensive insulin therapy (continuous subcutaneous insulin infusion [n = 12] or multiple daily injections [n = 18]) and then randomized to either pioglitazone (PIO group;45 mg/day), ramipril (RAM group; 10 mg/day), or placebo (PLC group) for 36 weeks. Euglycemic-hyperinsulinemic clamp was used to quantify insulin resistance, and plethysmography was used to assess changes in forearm blood flow (FBF) after 1) 5 min of reactive hyperemia and 2) brachial artery infusion of acetylcholine (7.5, 15, and 30 microg/min) and sodium nitroprusside (3 and 10 microg/min). RESULTS: The decreases in A1C (approximately 9.0-7.0%) and fasting plasma glucose (approximately 190-128 mg/dl) were equal in all groups. In the PIO group, glucose disposal increased from 3.1 to 4.7 mg x kg(-1) x min(-1), and there was a greater decrease in plasma triglycerides ( approximately 148 vs. 123 mg/dl) and free fatty acids (approximately 838 vs. 595 mEq/l) compared with the RAM or PLC groups (P < 0.05). Plasma adiponectin doubled with pioglitazone treatment (6.2 +/- 0.7 to 13.1 +/- 1.8 microg/ml), while endothelin-1 decreased only with ramipril treatment (2.5 +/- 0.2 to 1.1 +/- 0.2 pg/ml) (P < 001). The increase in FBF during reactive hyperemia (215%) and acetylcholine (from 132 to 205%, 216 to 262%, and 222 to 323%) was greater in the PIO versus RAM or PLC groups. In contrast, FBF during sodium nitroprusside treatment was greater in the RAM group (141-221% and 218-336%) compared with the PIO or PLC groups (all P < 0.05). CONCLUSIONS: Addition of pioglitazone or ramipril to intensive insulin therapy in type 2 diabetes further improves vascular dysfunction. Pioglitazone enhances endothelial-mediated vasodilation, whereas ACE inhibition enhances endothelial independent vasodilation. These different vascular effects, combined with the observation that pioglitazone decreases free fatty acids and triglycerides and increases adiponectin, while ramipril reduces endothelin-1, suggest that different mechanisms underlie the vascular responses. PMID- 17909085 TI - Effect of cinnamon on glucose control and lipid parameters. AB - OBJECTIVE: To perform a meta-analysis of randomized controlled trials of cinnamon to better characterize its impact on glucose and plasma lipids. RESEARCH DESIGN AND METHODS: A systematic literature search through July 2007 was conducted to identify randomized placebo-controlled trials of cinnamon that reported data on A1C, fasting blood glucose (FBG), or lipid parameters. The mean change in each study end point from baseline was treated as a continuous variable, and the weighted mean difference was calculated as the difference between the mean value in the treatment and control groups. A random-effects model was used. RESULTS: Five prospective randomized controlled trials (n = 282) were identified. Upon meta-analysis, the use of cinnamon did not significantly alter A1C, FBG, or lipid parameters. Subgroup and sensitivity analyses did not significantly change the results. CONCLUSIONS: Cinnamon does not appear to improve A1C, FBG, or lipid parameters in patients with type 1 or type 2 diabetes. PMID- 17909086 TI - Oral glucose tolerance test: a reliable tool for early detection of glucose abnormalities in patients with acute myocardial infarction in clinical practice: a report on repeated oral glucose tolerance tests from the GAMI study. AB - OBJECTIVE: Previously undetected glucose abnormalities are common in patients with acute myocardial infarction (AMI). We evaluated long-term reliability of early glucometabolic classification of patients with AMI by repeated oral glucose tolerance tests (OGTTs). RESEARCH DESIGN AND METHODS: A glucometabolic OGTT-based classification was obtained in 122 patients by measuring capillary whole-blood glucose. The classification was performed on three occasions, before hospital discharge and 3 and 12 months thereafter. RESULTS: At discharge, 34, 31, and 34% were classified as having normal glucose tolerance, impaired glucose tolerance (IGT), or type 2 diabetes, respectively, and 93% of all patients with type 2 diabetes were still classified with type 2 diabetes (n = 27) or IGT (n = 12) after 12 months. The agreements between the OGTTs at discharge and 3 and 12 months were kappa = 0.35, P < 0.001, and kappa = 0.43, P < 0.001, respectively. CONCLUSIONS: The outcome of an OGTT performed in AMI patients at hospital discharge reliably informs on long-term glucometabolic state. PMID- 17909087 TI - The dipeptidyl peptidase-4 inhibitor vildagliptin improves beta-cell function and insulin sensitivity in subjects with impaired fasting glucose. AB - OBJECTIVE: To evaluate the effect of treatment with the dipeptidyl peptidase (DPP)-4 inhibitor vildagliptin on insulin sensitivity and beta-cell function in subjects with impaired fasting glucose (IFG). RESEARCH DESIGN AND METHODS: A total of 22 subjects with IFG (11 female and 11 male, mean +/- SD age 59.6 +/- 11.5 years) were treated orally with 100 mg vildagliptin once daily in a single blind study. Subjects received placebo for 2 weeks (run-in) followed by vildagliptin for 6 weeks (treatment) and then placebo for 2 weeks (washout). A frequently sampled intravenous glucose tolerance test (FSIGT), followed by a 2-h meal tolerance test (MTT), was performed at 2, 8, and 10 weeks. From the FSIGT, the acute insulin response to glucose (AIR(g)) and insulin sensitivity index (S(I)) were determined and used to compute the disposition index (AIR(g) x S(I)) as a measure of beta-cell function. RESULTS: Fasting plasma glucose did not change after 6 weeks of vildagliptin treatment. With treatment, mean +/- SEM AIR(g) increased from 224 +/- 44 to 286 +/- 52 pmol/l (P < 0.05), and S(I) improved from 2.8 +/- 0.5 to 3.5 +/- 0.5 x 10(-5) x min(-1) x pmol(-1) x l (P < 0.01), resulting in an increase in the disposition index from 688 +/- 180 to 1,164 +/- 318 x 10(-5)/min (P < 0.05). These effects were not sustained after washout. During the MTT, the incremental area under the glucose curve was significantly decreased after treatment (240 +/- 15 vs. 191 +/- 14 mmol x l(-1) x min(-1); P = 0.002), but this effect was not sustained after washout. CONCLUSIONS: The DPP-4 inhibitor vildagliptin improves insulin sensitivity and beta-cell function, leading to improved postprandial glycemia in subjects with IFG, who are known to have beta-cell dysfunction. Thus, vildagliptin may prevent progression to diabetes in high-risk subjects. PMID- 17909088 TI - Relationship of prospective GHb to glycated serum proteins in incident diabetic retinopathy: implications of the glycation gap for mechanism of risk prediction. PMID- 17909089 TI - Ten-year trends in self-rated health among Spanish adults with diabetes, 1993 2003. AB - OBJECTIVE: Improving health-related quality of life among individuals with diabetes is a public health goal. This study sought to assess trends in self rated "fair" and "poor" health among Spanish adults with diabetes and to identify factors associated with fair and poor health using data from all five National Health Surveys conducted from 1993 to 2003. RESEARCH DESIGN AND METHODS: Poisson regression analysis was used to assess the trend in fair or poor health status during the period 1993-2003 and to compare prevalences between diabetes and nondiabetes sufferers. RESULTS: The most relevant finding of this study is that, among Spanish adults with diabetes, prevalence of fair or poor heath is more than double that of individuals without diabetes and that overall prevalence did not vary during the decade 1993-2003 (71.2 to 70.5%). The variables associated with an increased risk of self-rated fair or poor health were as follows: age 54-64 years (odds ratio [OR] 1.5) or >or= 65years (2.1), presence of comorbidity (4.3), female sex (1.2), lower educational level (1.7), obesity (1.3), and no physical activity (1.6). CONCLUSIONS: The lack of improvement in self-rated health among Spanish adults with diabetes calls for urgent implementation of health-promotion, prevention, and diabetes-management strategies aimed at enhancing the quality of life of such individuals. PMID- 17909090 TI - Head-to-head comparison of sirolimus- and paclitaxel-eluting stent in the same diabetic patient with multiple coronary artery lesions: a prospective, randomized, multicenter study. AB - OBJECTIVE: It is still controversial whether sirolimus-eluting stent (SES) and paclitaxel-eluting stent (PES) are equally effective in patients with diabetes. In these patients, multiple individual variables may be responsible for neointimal hyperplasia, thus making difficult the comparison of the two drug eluting stents (DES). RESEARCH DESIGN AND METHODS: We designed a prospective, randomized study to compare the efficacy in prevention of restenosis of SES and PES, both implanted in the same diabetic patient with multiple de novo coronary artery lesions undergoing elective percutaneous coronary intervention. We enrolled 60 patients with diabetes with at least two significant de novo angiographic stenoses in different coronary segments. The primary end point was in-stent late luminal loss (LLL) at 8-month angiographic follow-up. RESULTS: A total of 120 lesions were successfully treated with the randomly assigned DES (SES, n = 60; PES, n = 60). In-stent LLL was lower in the SES than in the PES group (0.26 +/- 0.4 vs. 0.50 +/- 0.6 mm; P = 0.01). Coronary lesions treated with SES presented a reduced in-stent LLL in 40 (68%) patients, while PES resulted in a lower in-stent LLL in 19 (32%) patients (P = 0.0002). At multivariable analysis, the type of DES implanted was the only independent predictor of in stent LLL (odds ratio 2.3 [95% CI 1.1-5.0]; P = 0.03). CONCLUSIONS: SES directly compared with PES in the same diabetic patient is associated with a decrease in the extent of in-stent LLL at 8 months, suggesting a reduced risk of restenosis. PMID- 17909091 TI - Hypoglycemia in pregnant women with type 1 diabetes: predictors and role of metabolic control. AB - OBJECTIVE: In pregnancy with type 1 diabetes, we evaluated occurrence of mild and severe hypoglycemia and analyzed the influence of strict metabolic control, nausea, vomiting, and other potential predictors of occurrence of severe hypoglycemia. RESEARCH DESIGN AND METHODS: A prospective observational study of 108 consecutive pregnant women with type 1 diabetes was conducted. At 8, 14, 21, 27, and 33 weeks of gestation, patients performed self-monitored plasma glucose (SMPG) (eight/day) for 3 days and completed a questionnaire on nausea, vomiting, hypoglycemia awareness, and history of mild (managed by the patient) and severe (requiring assistance from others) hypoglycemia. RESULTS: Forty-nine (45%) women experienced 178 severe hypoglycemic events, corresponding to 5.3, 2.4, and 0.5 events/patient-year in the first, second, and third trimesters, respectively. The incidence of mild hypoglycemia was 5.5 events/patient-week in early pregnancy and decreased throughout pregnancy (P < 0.0001), regardless of presence of severe hypoglycemia. Prevalence of nausea and vomiting, mild hypoglycemia, and fraction of SMPG readings or=18 years. Fasting plasma glucose and serum ferritin were measured. Dietary information was collected by 3-day food records. RESULTS: Serum ferritin was associated with elevated risk of diabetes even adjusted for age, sex, nondietary factors, and dietary factors. No association among total iron intake, nonheme iron intake, and diabetes risk was found. However, higher heme iron intake was significantly associated with elevated risk of diabetes after adjusting for known factors. CONCLUSIONS: In Chinese, associations among higher serum ferritin level, higher heme iron intake, and elevated risk of diabetes were found. PMID- 17909093 TI - Clinical experience with the addition of pramlintide in patients with insulin requiring type 2 diabetes. PMID- 17909094 TI - Neuritin mediates nerve growth factor-induced axonal regeneration and is deficient in experimental diabetic neuropathy. AB - OBJECTIVE: Axonal regeneration is defective in both experimental and clinical diabetic neuropathy, contributing to loss of axonal extremities and neuronal dysfunction. The mechanisms behind this failure are not fully understood; however, a deficit in neurotrophic support and signaling has been implicated. RESEARCH DESIGN AND METHODS: We investigated the expression of neuritin (also known as candidate plasticity gene 15, cpg15) in the sensory nervous system of control rats and rats with streptozotocin (STZ)-induced diabetes using microarray PCR, Western blotting, and immunocytochemical analysis. The functional role of neuritin in sensory neurons in vitro was assessed using silencing RNA. RESULTS: Neuritin was expressed by a population of small-diameter neurons in the dorsal root ganglia (DRG) and was anterogradely and retrogradely transported along the sciatic nerve in vivo. Nerve growth factor (NGF) treatment induced an increase in the transcription and translation of neuritin in sensory neurons in vitro. This increase was both time and dose dependent and occurred via mitogen-activated protein kinase or phosphatidylinositol-3 kinase activation. Inhibition of neuritin using silencing RNA abolished NGF-mediated neurite outgrowth, demonstrating the crucial role played by neuritin in mediating regeneration. Neuritin levels were reduced in both the DRG and sciatic nerve of rats with 12 weeks of STZ-induced diabetes, and these deficits were reversed in vivo by treatment with NGF. CONCLUSIONS: Manipulation of neuritin levels in diabetes may therefore provide a potential target for therapeutic intervention in the management of neuropathy. PMID- 17909095 TI - In vivo evidence for inverse agonism of Agouti-related peptide in the central nervous system of proopiomelanocortin-deficient mice. AB - OBJECTIVE: Melanocyte-stimulating hormone (MSH) peptides processed from proopiomelanocortin (POMC) regulate energy homeostasis by activating neuronal melanocortin receptor (MC-R) signaling. Agouti-related peptide (AgRP) is a naturally occurring MC-R antagonist but also displays inverse agonism at constitutively active melanocortin-4 receptor (MC4-R) expressed on transfected cells. We investigated whether AgRP functions similarly in vivo using mouse models that lack all neuronal MSH, thereby precluding competitive antagonism of MC-R by AgRP. RESEARCH DESIGN AND METHODS: Feeding and metabolic effects of the MC-R agonist melanotan II (MTII), AgRP, and ghrelin were investigated after intracerebroventricular injection in neural-specific POMC-deficient (Pomc(-/ )Tg/+) and global POMC-deficient (Pomc(-/-)) mice. Gene expression was quantified by RT-PCR. RESULTS: Hyperphagic POMC-deficient mice were more sensitive than wild type mice to the anorectic effects of MTII. Hypothalamic melanocortin-3 (MC3)/4-R mRNAs in POMC-deficient mice were unchanged, suggesting increased receptor sensitivity as a possible mechanism for the heightened anorexia. AgRP reversed MTII-induced anorexia in both mutant strains, demonstrating its ability to antagonize MSH agonists at central MC3/4-R, but did not produce an acute orexigenic response by itself. The action of ghrelin was attenuated in Pomc(-/ )Tg/+ mice, suggesting decreased sensitivity to additional orexigenic signals. However, AgRP induced delayed and long-lasting modifications of energy balance in Pomc(-/-)Tg/+, but not glucocorticoid-deficient Pomc(-/-) mice, by decreasing oxygen consumption, increasing the respiratory exchange ratio, and increasing food intake. CONCLUSIONS: These data demonstrate that AgRP can modulate energy balance via a mechanism independent of MSH and MC3/4-R competitive antagonism, consistent with either inverse agonist activity at MC-R or interaction with a distinct receptor. PMID- 17909096 TI - Stromal cell-derived factor-1/CXCL12 stimulates chemorepulsion of NOD/LtJ T-cell adhesion to islet microvascular endothelium. AB - OBJECTIVE: Diabetogenic T-cell recruitment into pancreatic islets facilitates beta-cell destruction during autoimmune diabetes, yet specific mechanisms governing this process are poorly understood. The chemokine stromal cell-derived factor-1 (SDF-1) controls T-cell recruitment, and genetic polymorphisms of SDF-1 are associated with early development of type 1 diabetes. RESEARCH DESIGN AND METHODS: Here, we examined the role of SDF-1 regulation of diabetogenic T-cell adhesion to islet microvascular endothelium. Islet microvascular endothelial cell monolayers were activated with tumor necrosis factor-alpha (TNF-alpha), subsequently coated with varying concentrations of SDF-1 (1-100 ng/ml), and assayed for T-cell/endothelial cell interactions under physiological flow conditions. RESULTS: TNF-alpha significantly increased NOD/LtJ T-cell adhesion, which was completely blocked by SDF-1 in a dose-dependent manner, revealing a novel chemorepulsive effect. Conversely, SDF-1 enhanced C57BL/6J T-cell adhesion to TNF-alpha-activated islet endothelium, demonstrating that SDF-1 augments normal T-cell adhesion. SDF-1 chemorepulsion of NOD/LtJ T-cell adhesion was completely reversed by blocking G(i)alpha-protein-coupled receptor activity with pertussis toxin. CXCR4 protein expression was significantly decreased in NOD/LtJ T-cells, and inhibition of CXCR4 activity significantly reversed SDF-1 chemorepulsive effects. Interestingly, SDF-1 treatment significantly abolished T cell resistance to shear-mediated detachment without altering adhesion molecule expression, thus demonstrating decreased integrin affinity and avidity. CONCLUSIONS: In this study, we have identified a previously unknown novel function of SDF-1 in negatively regulating NOD/LtJ diabetogenic T-cell adhesion, which may be important in regulating diabetogenic T-cell recruitment into islets. PMID- 17909097 TI - Metformin inhibits hepatic gluconeogenesis through AMP-activated protein kinase dependent regulation of the orphan nuclear receptor SHP. AB - OBJECTIVE: Metformin is an antidiabetic drug commonly used to treat type 2 diabetes. The aim of the study was to determine whether metformin regulates hepatic gluconeogenesis through the orphan nuclear receptor small heterodimer partner (SHP; NR0B2). RESEARCH DESIGN AND METHODS: We assessed the regulation of hepatic SHP gene expression by Northern blot analysis with metformin and adenovirus containing a constitutive active form of AMP-activated protein kinase (AMPK) (Ad-AMPK) and evaluated SHP, PEPCK, and G6Pase promoter activities via transient transfection assays in hepatocytes. Knockdown of SHP using siRNA SHP was conducted to characterize the metformin-induced inhibition of hepatic gluconeogenic gene expression in hepatocytes, and metformin-and adenovirus SHP (Ad-SHP)-mediated hepatic glucose production was measured in B6-Lep(ob/ob) mice. RESULTS: Hepatic SHP gene expression was induced by metformin, 5-aminoimidazole-4 carboxamide-1-beta-d-ribofuranoside (AICAR), and Ad-AMPK. Metformin-induced SHP gene expression was abolished by adenovirus containing the dominant negative form of AMPK (Ad-DN-AMPK), as well as by compound C. Metformin inhibited hepatocyte nuclear factor-4alpha-or FoxA2-mediated promoter activity of PEPCK and G6Pase, and the inhibition was blocked with siRNA SHP. Additionally, SHP knockdown by adenovirus containing siRNA SHP inhibited metformin-mediated repression of cAMP/dexamethasone-induced hepatic gluconeogenic gene expression. Furthermore, oral administration of metformin increased SHP mRNA levels in B6-Lep(ob/ob) mice. Overexpression of SHP by Ad-SHP decreased blood glucose levels and hepatic gluconeogenic gene expression in B6-Lep(ob/ob) mice. CONCLUSIONS: We have concluded that metformin inhibits hepatic gluconeogenesis through AMPK-dependent regulation of SHP. PMID- 17909098 TI - Expression of NG,NG-dimethylarginine dimethylaminohydrolase and protein arginine N-methyltransferase isoforms in diabetic rat kidney: effects of angiotensin II receptor blockers. AB - OBJECTIVE: The nitric oxide (NO) synthase inhibitor asymmetric dimethylarginine (ADMA) is generated by protein arginine N-methyltransferase (PRMT)-1 and is metabolized by N(G),N(G)-dimethylarginine dimethylaminohydrolase (DDAH). We tested the hypothesis that increased serum ADMA (S(ADMA)) in the streptozotocin (STZ)-induced diabetic rat model of diabetes is mediated by an angiotensin receptor blocker-sensitive change in DDAH or PRMT expression. RESEARCH DESIGN AND METHODS: Data were compared from four groups of rats: sham-injected controls, untreated STZ-induced diabetic rats at 4 weeks, STZ-induced diabetic rats administered the angiotensin II (Ang II) receptor blocker telmisartan for 2 weeks, and control rats administered telmisartan for 2 weeks. RESULTS: Immunostaining and Western blotting of microdissected nephron segments localized DDAH I in the proximal tubules and DDAH II in the glomeruli, afferent arterioles, macula densa, and distal nephron. Renal Ang II and S(ADMA) increased with diabetes but were normalized by 2 weeks of telmisartan. DDAH I expression was decreased in diabetic kidneys, while DDAH II expression was increased. These changes were reversed by telmisartan, which also reduced expression of PRMT-1 and -5. Telmisartan increased expressions of DDAH I but decreased DDAH II in Ang II stimulated kidney slices ex vivo. CONCLUSIONS: Renal Ang II and S(ADMA) are increased in insulinopenic diabetes. They are normalized by an Ang II receptor blocker, which increases the renal expression of DDAH I, decreases PRMT-1, and increases renal NO metabolites. PMID- 17909099 TI - TCF7L2 is not a major susceptibility gene for type 2 diabetes in Pima Indians: analysis of 3,501 individuals. AB - OBJECTIVE: The transcription factor 7-like 2 (TCF7L2) gene was initially reported to be associated with type 2 diabetes in Icelandic, Danish, and U.S. populations. We investigated whether TCF7L2 also has a role in type 2 diabetes susceptibility in Pima Indians. RESEARCH DESIGN AND METHODS: The six variants reported to be associated with type 2 diabetes in the Icelandic study were genotyped in a population-based sample of 3,501 Pima Indians (1,561 subjects had type 2 diabetes, and 1,940 did not have diabetes). In addition, the coding and promoter regions of TCF7L2 were sequenced in 24 Pima subjects. The one variant identified by sequencing, 35 additional database variants positioned in introns, and the six variants reported in the Icelandic study were genotyped in Pima families to determine the haplotype structure of TCF7L2 among Pima Indians. Fourteen representative variants were selected and genotyped in 3,501 Pima Indians. RESULTS: The six variants initially reported to be associated with type 2 diabetes were less common in Pima Indians compared with samples of European origin, and none were associated with type 2 diabetes. One representative variant, rs1225404, was nominally associated with type 2 diabetes in a general model (additive P = 0.03, dominant P = 0.005) but not in a within-family analysis (additive P = 0.2, dominant P = 0.07). However, several variants were associated with BMI; in particular, rs12255372 was associated in both general and within family analyses (both P = 0.0007). Modest associations were also found with traits predictive for type 2 diabetes. CONCLUSIONS: Variation within TCF7L2 does not confer major risk for type 2 diabetes among the Pima Indian population. PMID- 17909101 TI - The effect of the NMDA receptor antagonist MK-801 on the acquisition and extinction of learned fear in the developing rat. AB - Recent findings reveal qualitative developmental differences in extinction of learned fear. The present study explored potential developmental differences in the role of NMDA in acquisition and extinction. Rats were injected with MK-801 prior to fear conditioning or extinction training. Acquisition was found to be NMDA dependent in both age groups, whereas extinction was found to be NMDA dependent in 23-day-old rats, but NMDA independent in 16-day-old rats. These results illustrate another fundamental developmental difference in extinction as well as a dissociation in the role of NMDA in the acquisition and extinction of fear early in development. PMID- 17909100 TI - The extracellular protease matrix metalloproteinase-9 is activated by inhibitory avoidance learning and required for long-term memory. AB - Matrix metalloproteinases (MMPs) are a family of extracellularly acting proteolytic enzymes with well-recognized roles in plasticity and remodeling of synaptic circuits during brain development and following brain injury. However, it is now becoming increasingly apparent that MMPs also function in normal, nonpathological synaptic plasticity of the kind that may underlie learning and memory. Here, we extend this idea by investigating the role and regulation of MMP 9 in an inhibitory avoidance (IA) learning and memory task. We demonstrate that following IA training, protein levels and proteolytic activity of MMP-9 become elevated in hippocampus by 6 h, peak at 12-24 h, then decline to baseline values by approximately 72 h. When MMP function is abrogated by intrahippocampal infusion of a potent gelatinase (MMP-2 and MMP-9) inhibitor 3.5 h following IA training, a time prior to the onset of training-induced elevation in levels, IA memory retention is significantly diminished when tested 1-3 d later. Animals impaired at 3 d exhibit robust IA memory when retrained, suggesting that such impairment is not likely attributed to toxic or other deleterious effects that permanently disrupt hippocampal function. In anesthetized adult rats, the effective distance over which synaptic plasticity is impaired by a single intrahippocampal infusion of the MMP inhibitor of the kind that blocks IA memory is approximately 1200 microm. Taken together, these data suggest that IA training induces a slowly emerging, but subsequently protracted period of MMP-mediated proteolysis critical for enabling long-lasting synaptic modification that underlies long-term memory consolidation. PMID- 17909102 TI - Fibroblast growth factor 1 gene and hypertension: from the quantitative trait locus to positional analysis. AB - BACKGROUND: The distal portion of the long arm of chromosome 5 is linked to hypertension and contains functional candidate blood pressure-regulating genes. METHODS AND RESULTS: Tightening the grid of microsatellite markers under this quantitative trait locus in the Silesian Hypertension Study (629 individuals from 207 Polish hypertensive families) provided enhanced support for linkage of this region to blood pressure (maximal Z=3.51, P=0.0002). The fine mapping, comparative genomics, and functional prioritization identified fibroblast growth factor 1 gene (FGF1) as the positional candidate. Linkage disequilibrium mapping based on 51 single nucleotide polymorphisms spanning the locus showed no overlap between 3 independent haploblocks of FGF1 and the adjacent extragenic chromosomal regions. Single and multilocus family-based analysis revealed that genetic variation within FGF1 haploblock 1 was associated with hypertension and identified a common intronic single nucleotide polymorphism, rs152524, as the major driver of this association (P=0.0026). Real-time quantitative polymerase chain reaction and Western blotting analysis of renal tissue obtained from subjects undergoing unilateral nephrectomy showed an increase in both mRNA and protein FGF1 expression in hypertensive patients compared with normotensive controls. Renal immunohistochemistry revealed that FGF1 was expressed exclusively within the glomerular endothelial and mesangial cells. CONCLUSIONS: Our data demonstrate that genetic variation within FGF1 cosegregates with elevated blood pressure in hypertensive families and that this association is likely to be mediated by upregulation of renal FGF1 expression. The results of our study will need to be replicated in other cohorts. PMID- 17909103 TI - Persistent cardiac troponin I elevation in stabilized patients after an episode of acute coronary syndrome predicts long-term mortality. AB - BACKGROUND: In patients with non-ST-elevation acute coronary syndrome, any troponin elevation is associated with an increased risk for cardiovascular events. However, the prevalence and prognostic importance of persistent troponin elevation in stabilized patients after an episode of non-ST-elevation acute coronary syndrome are unknown and were therefore assessed in this study. METHODS AND RESULTS: Cardiac troponin I (cTnI) was measured in 1092 stabilized patients at 6 weeks and 3 and 6 months after enrollment in the FRagmin and Fast Revascularization during InStability in Coronary artery disease (FRISC-II) trial. cTnI was analyzed with the Access AccuTnI assay with the application of different prognostic cutoffs. Outcomes were assessed through 5 years. Elevated cTnI levels >0.01 microg/L were found in 48% of the study patients at 6 weeks, in 36% at 6 months, and in 26% at all 3 measurements. cTnI elevation was associated with increased age and other cardiovascular high-risk features. The lowest tested cTnI cutoff (0.01 microg/L) was prognostically most useful and was independently predictive of mortality (hazard ratio, 2.1 [95% confidence interval, 1.3 to 3.3]; P=0.001) on multivariable analysis adjusted for cardiovascular risk factors and randomization to an invasive versus noninvasive treatment strategy, whereas it was related to myocardial infarction only on univariate analysis. CONCLUSIONS: Persistent minor cTnI elevation can be detected frequently in patients stabilized after an episode of non-ST-elevation acute coronary syndrome with the use of a sensitive assay. Elevated cTnI levels >0.01 microg/L predict mortality during long-term follow-up. Our results emphasize the importance of further troponin testing in non-ST-elevation acute coronary syndrome patients after hospital discharge. PMID- 17909104 TI - CC chemokine ligand-5 (CCL5/RANTES) and CC chemokine ligand-18 (CCL18/PARC) are specific markers of refractory unstable angina pectoris and are transiently raised during severe ischemic symptoms. AB - BACKGROUND: Chemokines play an important role in atherogenesis and in ischemic injury and repair; however, prospective data on individual chemokines in unstable angina pectoris (UAP) are scarce. Therefore, we assessed chemokine patterns in a prospective cohort of patients with UAP. METHODS AND RESULTS: Plasma samples of 54 patients with Braunwald class IIIB UAP were examined at baseline for 11 chemokines and 5 inflammatory mediators via multiplex analysis. Levels of CC chemokine ligand (CCL)-5 (also known as RANTES [regulated on activation, normally T-cell expressed, and secreted]; 32.7 versus 23.1 ng/mL, P=0.018) and CCL18 (also known as PARC [pulmonary and activation-regulated chemokine]; 104.4 versus 53.7 ng/mL, P=0.011) were significantly elevated in patients with refractory ischemic symptoms versus stabilized patients. Temporal monitoring by ELISA of CCL5, CCL18, and soluble CD40 ligand (sCD40) levels revealed a drop in CCL5 and sCD40L levels in all UAP patients from day 2 onward (CCL5 12.1 ng/mL, P<0.001; sCD40L 1.35 ng/mL, P<0.05), whereas elevated CCL18 levels were sustained for at least 2 days, then were decreased at 180 days after inclusion (34.5 ng/mL, P<0.001). Peripheral blood mononuclear cells showed increased protein expression of chemokine receptors CCR3 and CCR5 in CD3+ and CD14+ cells at baseline compared with 180 days after inclusion, whereas mRNA levels were downregulated, which was attributable in part to a postischemic release of human neutrophil peptide-3 positive neutrophils and in part to negative feedback. Finally, elevated CCL5 and CCL18 levels predicted future cardiovascular adverse events, whereas C-reactive protein and sCD40L levels did not. CONCLUSIONS: We are the first to report that CCL18 and CCL5 are transiently raised during episodes of UAP, and peak levels of both chemokines are indicative of refractory symptoms. Because levels of both chemokines, as well as of cognate receptor expression by circulating peripheral blood mononuclear cells, are increased during cardiac ischemia, this may point to an involvement of CCL5/CCL18 in the pathophysiology of UAP and/or post-UAP responses. PMID- 17909105 TI - A secretion trap screen in yeast identifies protease inhibitor 16 as a novel antihypertrophic protein secreted from the heart. AB - BACKGROUND: Cardiomyocyte hypertrophy is of central importance in the development of congestive heart failure. Whether proteins secreted from the myocardium itself contribute to myocardial hypertrophy is largely unknown. METHODS AND RESULTS: We performed a genetic yeast secretion trap screen using a murine cardiac cDNA library and identified 54 cardiac proteins that contained a secretion signal. When determining their mRNA expression in the myocardium of failing hearts, we found protease inhibitor 16 (PI16) to be strongly upregulated in hypertrophic and failing myocardium. PI16, a 489-amino acid protein with an unknown function, also displayed enhanced expression on the protein level after serum stimulation of primary cardiomyocytes and in failing myocardium. We found PI16 to be secreted rapidly by primary cardiomyocytes into the culture medium, where it inhibited cardiomyocyte growth. RNA interference-mediated suppression of endogenous PI16 in primary cardiomyocytes significantly enhanced cardiomyocyte size. Transgenic mice overexpressing PI16 in a cardiomyocyte-specific manner showed normal cardiac function but had smaller hearts with hypotrophic cardiomyocytes. CONCLUSIONS: Taken together, we identified 54 putatively secreted cardiac proteins. PI16, a novel protein secreted from the heart, is strongly upregulated early in heart failure and inhibits growth of cardiomyocytes both in vitro and in vivo. PI16 might represent a novel therapeutic target in heart failure. PMID- 17909106 TI - Identification of a novel role of T cells in postnatal vasculogenesis: characterization of endothelial progenitor cell colonies. AB - BACKGROUND: The colony number of early endothelial progenitor cells (EPCs) has been used as a quantitative indicator of the number of EPCs in the blood or as a biological marker of cardiovascular diseases. In the present study, we found a subset of T cells that were localized at the center of the EPC colony and played a pivotal role in colony formation and differentiation of early EPCs. METHODS AND RESULTS: We found that CD3+ CD31+ CXCR4+ T cells (referred to as angiogenic T cells in the present study) constituted the center of EPC colonies during cultures of human peripheral blood mononuclear cells. These angiogenic T cells were required for colony formation and differentiation of early EPCs. They secreted high levels of angiogenic cytokines such as vascular endothelial growth factor, interleukin-8, and matrix metalloproteinases. Angiogenic T cells showed superior angiogenic potential to the other subset of T cells in the experiments with regard to Matrigel tube formation, adhesion, transendothelial migration, and collagen invasion assay, mainly through the stromal cell-derived factor 1/CXCR-4 axis. Furthermore, angiogenic T cells enhanced endothelial cell proliferation and function. In vivo study showed that angiogenic T cells play an important role in the process of vessel formation. Clinical study showed that the level of angiogenic T cells in the peripheral blood was well correlated with EPC colony numbers and had inverse relationships with age and the number of risk factors for coronary artery disease. CONCLUSIONS: These findings suggest that angiogenic T cells could be a potential therapeutic target for ischemic cardiovascular diseases. PMID- 17909107 TI - CD39/ectonucleoside triphosphate diphosphohydrolase 1 provides myocardial protection during cardiac ischemia/reperfusion injury. AB - BACKGROUND: Extracellular adenosine, generated from extracellular nucleotides via ectonucleotidases, binds to specific receptors and provides cardioprotection from ischemia and reperfusion. In the present study, we studied ecto-enzymatic ATP/ADP phosphohydrolysis by select members of the ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) family during myocardial ischemia. METHODS AND RESULTS: As a first step, we used a murine model of myocardial ischemia and in situ preconditioning and performed pharmacological studies with polyoxometalate 1, a potent E-NTPDase inhibitor (Na6[H2W12O40]). Polyoxometalate 1 treatment increased infarct sizes and abolished beneficial effects of preconditioning. To define relative contributions of distinct E-NTPDases, we investigated transcriptional responses of E-NTPDases 1 to 3 and 8 to preconditioning. We noted robust and selective induction of E-NTPDase 1 (CD39) transcript and protein. Histological analysis of preconditioned myocardium localized CD39 induction to endothelia and myocytes. Cd39-/- mice exhibited larger infarct sizes with ischemia (cd39+/+ 43.0+/-3.3% versus cd39-/- 52%+/-1.8; P<0.05), and cardioprotection was abrogated by preconditioning (cd39+/+ 13.3%+/-1.5 versus cd39-/- 50.5%+/-2.8; P<0.01). Heightened levels of injury after myocardial ischemia and negligible preconditioning benefits in cd39-/- mice were corrected by infusion of the metabolic product (AMP) or apyrase. Moreover, apyrase treatment of wild-type mice resulted in 43+/-4.2% infarct size reduction (P<0.01). CONCLUSIONS: Taken together, these studies reveal E-NTPDase 1 in cardioprotection and suggest apyrase in the treatment of myocardial ischemia. PMID- 17909109 TI - Viewpoint: Maria Grazia Modena MD, FESC. Interview by Barry Shurlock. PMID- 17909108 TI - CXCR6 promotes atherosclerosis by supporting T-cell homing, interferon-gamma production, and macrophage accumulation in the aortic wall. AB - BACKGROUND: T lymphocytes are thought to be important in atherosclerosis, but very little is known about the mechanisms of lymphocyte recruitment into atherosclerosis-prone aortas. In this study we tested the hypothesis that CXCR6, a chemokine receptor that is expressed on a subset of CD4+ T helper 1 cells and natural killer T cells, is involved in lymphocyte homing into the aortic wall and modulates the development and progression of atherosclerosis. METHODS AND RESULTS: To investigate the role of CXCR6 in the development and progression of atherosclerosis, we bred CXCR6-deficient (CXCR6(GFP/GFP)) mice with apolipoprotein E-deficient (ApoE(-/-)) mice. We found that CXCR6(GFP/GFP)/ApoE(-/ ) mice fed a Western diet for 17 weeks or a chow diet for 56 weeks had decreased atherosclerosis compared with ApoE(-/-) controls. Flow cytometry analysis of the aortas from CXCR6(GFP/GFP)/ApoE(-/-) mice showed that the reduction of atherosclerosis was accompanied by a decreased percentage of CXCR6+ T cells within the aortas. Short-term homing experiments demonstrated that CXCR6 is involved in the recruitment of CXCR6+ leukocytes into the atherosclerosis-prone aortic wall. The reduced percentage of CXCR6+ T cells within the aortas resulted in significantly diminished production of interferon-gamma and reduction of CD11b+/CD68+ macrophages in the aorta. CONCLUSIONS: These data provide evidence for a proatherosclerotic role of CXCR6. Absence of CXCR6 alters the recruitment of CXCR6+ leukocytes and modulates the local immune response within the aortic wall. PMID- 17909111 TI - Images in cardiovascular medicine. Optical coherence tomography in the setting of an acute anterior myocardial infarction. PMID- 17909110 TI - Images in cardiovascular medicine. Pseudoaneurysm and intracardiac fistula caused by an infected paclitaxel-eluting coronary stent. PMID- 17909112 TI - Letter by Weidemann et al regarding article, "Global diastolic strain rate for the assessment of left ventricular relaxation and filling pressure". PMID- 17909113 TI - Can we predict and prevent the onset of acute decompensated heart failure? PMID- 17909114 TI - Eat your fruits and vegetables but hold the salt. PMID- 17909115 TI - Carotid stents: unleashed, unproven. PMID- 17909116 TI - The argument to support broader application of extracranial carotid artery stent technology. PMID- 17909117 TI - Peroxisome proliferator-activated receptor alpha: friend or foe? PMID- 17909118 TI - Will the lessons from primary aldosteronism change the treatment of hypertension and left ventricular hypertrophy? PMID- 17909119 TI - Qualitative assessment of sympathetic neural drive in cardiometabolic disease: a new challenge. PMID- 17909120 TI - Differing pattern of sympathoexcitation in normal-weight and obesity-related hypertension. AB - Hypertension in normal-weight and obese individuals is characterized by activation of the sympathetic nervous system. Measurement of spillover of the sympathetic transmitter, norepinephrine, to plasma indicates that the regional pattern of sympathetic activation in the 2 "variants" of essential hypertension differs, excluding the heart in obesity-related hypertension. Whether sympathetic nerve firing characteristics also differ is unknown. We studied multiunit and single fiber sympathetic nerve firing properties in patients with normal-weight hypertension and obesity-related hypertension, comparing these with nerve characteristics in normal-weight and obese people with normal blood pressure. Both normal-weight hypertensive (n=10) and obese hypertensive (n=14) patients had increased total multiunit muscle sympathetic nerve activity compared with the normal-weight (n=11) and obese (n=11) people with normal blood pressure (65+/-4 versus 47+/-6 bursts per 100 heartbeats, P<0.01 in the normal-weight groups and 68+/-4 versus 53+/-3 bursts per 100 beats, P<0.01 in the obese groups). Sympathetic activation in normal-weight hypertension was characterized by increased firing rate of single vasoconstrictor fibers (70+/-8 versus 28+/-3 spikes per 100 beats; P<0.001), increased firing probability per heartbeat (39+/ 3% versus 20+/-3%; P<0.001), and higher incidence of multiple spikes per heartbeat (30+/-4% versus 17+/-4%; P<0.05). Sympathetic activation in obesity related hypertension differed, involving recruitment of previously silent fibers, which fired at a normal rate. The pattern of sympathetic activation in normal weight and obesity-related hypertension differs in terms of both the firing characteristics of individual sympathetic fibers and the sympathetic outflows involved. The underlying central nervous system mechanism and the adverse consequences of the 2 modes of sympathetic activation may differ. PMID- 17909121 TI - Absence of peroxisome proliferator-activated receptor-alpha abolishes hypertension and attenuates atherosclerosis in the Tsukuba hypertensive mouse. AB - Peroxisome proliferator-activated receptor-alpha is widely distributed in the vasculature where it is believed to exert pleiotropic antiatherogenic effects. Its role in the regulation of blood pressure is still unresolved; however, some evidence suggests that it may affect the renin-angiotensin system. We investigated its role in angiotensin II-induced hypertension in the Tsukuba hypertensive mouse (THM). This is a model of hypertension and atherosclerosis because of high angiotensin II and aldosterone levels as a result of the transgenic expression of the entire human renin-angiotensin system. Making the THM animals deficient in Peroxisome proliferator-activated receptor-alpha (THM/PPARKO) totally abolished hypertension and myocardial hypertrophy. This was accompanied by a reduction in plasma human active renin in THM/PPARKO mice compared with THM animals from 3525+/-128 mU/L to 1910+/-750 mU/L (P<0.05) and by a normalization of serum aldosterone (1.6+/-0.29 nmol/L versus 3.4+/-0.69 nmol/L; P=0.003). In the THM/PPARKO mice, the extent of atherosclerosis at the aortic sinus after a 12-week period on an atherogenic diet was decreased by >80%. In addition, the spontaneous formation of foam cells from peritoneal macrophages, a blood pressure-independent event, was reduced by 92% in the THM/PPARKO mice, suggesting protection from the usual oxidative stress in these animals, possibly because of lower prevailing angiotensin II levels. Finally, chronic fenofibrate treatment further elevated blood pressure in THM animals but not in THM/PPARKO animals. Taken together, these data indicate that peroxisome proliferator activated receptor-alpha may regulate the renin-angiotensin system. They raise the possibility that its activation may aggravate hypertension and hasten atherosclerosis in the context of an activated renin-angiotensin system. PMID- 17909123 TI - Cognitive improvement after treatment with second-generation antipsychotic medications in first-episode schizophrenia: is it a practice effect? AB - CONTEXT: Cognitive impairment in schizophrenia is frequent, involves multiple domains, and is enduring. Numerous recent clinical trials have suggested that second-generation antipsychotic medications significantly enhance cognition in schizophrenia. However, none of these studies included healthy controls undergoing repeated testing to assess the possibility that improvements might reflect simple practice effects. OBJECTIVE: To report the results on cognition of a randomized comparison of 2 widely prescribed second-generation antipsychotic medications, olanzapine and risperidone, in patients with first-episode schizophrenia and a healthy control group. DESIGN: Randomized clinical trial. SETTING: Hospital-based research units. Patients A total of 104 participants with first-episode schizophrenia and 84 healthy controls. MAIN OUTCOME MEASURES: Cognitive assessment of all study participants occurred at baseline, 6 weeks later, and 16 weeks later. Neurocognitive tests included measures of working memory and attention, speed, motor function, episodic memory, and executive function. RESULTS: No differential drug effects were observed. Of 16 cognitive measures, 9 demonstrated improvement over time and only 2 demonstrated greater rates of change than those observed in the healthy control group undergoing repeated assessment. The composite effect size for cognitive change was 0.33 in the healthy control group (attributed to practice) and 0.36 in the patients with first-episode schizophrenia. Improvements in cognition in the first-episode schizophrenia group could not be accounted for by medication dose, demographic variables, or intellectual level. CONCLUSIONS: The cognitive improvements observed in the trial were consistent in magnitude with practice effects observed in healthy controls, suggesting that some of the improvements in cognition in the first-episode schizophrenia group may have been due to practice effects (ie, exposure, familiarity, and/or procedural learning). Our results also indicated that differential medication effects on cognition were small. We believe that these findings have important implications for drug discovery and the design of registration trials that attempt to demonstrate cognitive enhancement. PMID- 17909124 TI - A systematic review of mortality in schizophrenia: is the differential mortality gap worsening over time? AB - CONTEXT: Despite improvements in mental health services in recent decades, it is unclear whether the risk of mortality in schizophrenia has changed over time. OBJECTIVE: To explore the distribution of standardized mortality ratios (SMRs) for people with schizophrenia. DATA SOURCES: Broad search terms were used in MEDLINE, PsychINFO, Web of Science, and Google Scholar to identify all studies that investigated mortality in schizophrenia, published between January 1, 1980, and January 31, 2006. References were also identified from review articles, reference lists, and communication with authors. STUDY SELECTION: Population based studies that reported primary data on deaths in people with schizophrenia. DATA EXTRACTION: Operationalized criteria were used to extract key study features and mortality data. DATA SYNTHESIS: We examined the distribution of SMRs and pooled selected estimates using random-effects meta-analysis. We identified 37 articles drawn from 25 different nations. The median SMR for all persons for all cause mortality was 2.58 (10%-90% quantile, 1.18-5.76), with a corresponding random-effects pooled SMR of 2.50 (95% confidence interval, 2.18-2.43). No sex difference was detected. Suicide was associated with the highest SMR (12.86); however, most of the major causes-of-death categories were found to be elevated in people with schizophrenia. The SMRs for all-cause mortality have increased during recent decades (P = .03). CONCLUSIONS: With respect to mortality, a substantial gap exists between the health of people with schizophrenia and the general community. This differential mortality gap has worsened in recent decades. In light of the potential for second-generation antipsychotic medications to further adversely influence mortality rates in the decades to come, optimizing the general health of people with schizophrenia warrants urgent attention. PMID- 17909125 TI - The Treatment for Adolescents With Depression Study (TADS): long-term effectiveness and safety outcomes. AB - CONTEXT: The Treatment for Adolescents With Depression Study evaluates the effectiveness of fluoxetine hydrochloride therapy, cognitive behavior therapy (CBT), and their combination in adolescents with major depressive disorder. OBJECTIVE: To report effectiveness outcomes across 36 weeks of randomized treatment. DESIGN AND SETTING: Randomized, controlled trial conducted in 13 academic and community sites in the United States. Cognitive behavior and combination therapies were not masked, whereas administration of placebo and fluoxetine was double-blind through 12 weeks, after which treatments were unblinded. Patients assigned to placebo were treated openly after week 12, and the placebo group is not included in these analyses by design. PARTICIPANTS: Three hundred twenty-seven patients aged 12 to 17 years with a primary DSM-IV diagnosis of major depressive disorder. INTERVENTIONS: All treatments were administered per protocol. MAIN OUTCOME MEASURES: The primary dependent measures rated blind to treatment status by an independent evaluator were the Children's Depression Rating Scale-Revised total score and the response rate, defined as a Clinical Global Impressions-Improvement score of much or very much improved. RESULTS: Intention-to-treat analyses on the Children's Depression Rating Scale Revised identified a significant time x treatment interaction (P < .001). Rates of response were 73% for combination therapy, 62% for fluoxetine therapy, and 48% for CBT at week 12; 85% for combination therapy, 69% for fluoxetine therapy, and 65% for CBT at week 18; and 86% for combination therapy, 81% for fluoxetine therapy, and 81% for CBT at week 36. Suicidal ideation decreased with treatment, but less so with fluoxetine therapy than with combination therapy or CBT. Suicidal events were more common in patients receiving fluoxetine therapy (14.7%) than combination therapy (8.4%) or CBT (6.3%). CONCLUSIONS: In adolescents with moderate to severe depression, treatment with fluoxetine alone or in combination with CBT accelerates the response. Adding CBT to medication enhances the safety of medication. Taking benefits and harms into account, combined treatment appears superior to either monotherapy as a treatment for major depression in adolescents. PMID- 17909126 TI - Prospective effects of attention-deficit/hyperactivity disorder, conduct disorder, and sex on adolescent substance use and abuse. AB - CONTEXT: Attention-deficit/hyperactivity disorder (ADHD), an early manifestation of externalizing behavior, may identify children at high risk for later substance abuse. However, the ADHD-substance abuse relationship often disappears when co occurring conduct disorder (CD) is considered. OBJECTIVE: To determine whether there is a prospective relationship between ADHD and the initiation of substance use and disorders, and whether this relationship depends on the ADHD subtype (hyperactive/impulsive or inattentive), CD, or sex. DESIGN, SETTING, AND PARTICIPANTS: Dimensional and categorical measures of ADHD and CD were examined via logistic regression analyses in relation to subsequent initiation of tobacco, alcohol, and illicit drug use by 14 years of age and onset of substance use disorders by 18 years of age in a population-based sample of 11-year-old twins (760 female and 752 male twins) from the Minnesota Twin Family Study. MAIN OUTCOME MEASURES: Structured interviews were administered to adolescents and their mothers regarding substance use and to generate diagnoses. RESULTS: For boys and girls, hyperactivity/impulsivity predicted initiation of all types of substance use, nicotine dependence, and cannabis abuse/dependence (for all, P < .05), even when controlling for CD at 2 time points. By contrast, relationships between inattention and substance outcomes disappeared when hyperactivity/impulsivity and CD were controlled for, with the possible exception of nicotine dependence. A categorical diagnosis of ADHD significantly predicted tobacco and illicit drug use only (adjusted odds ratios, 2.01 and 2.82, respectively). A diagnosis of CD between 11 and 14 years of age was a powerful predictor of substance disorders by 18 years of age (all odds ratios, > 4.27). CONCLUSIONS: Hyperactivity/impulsivity predicts later substance problems, even after growth in later-emerging CD is considered, whereas inattention alone poses less risk. Even a single symptom of ADHD or CD is associated with increased risk. Failure in previous research to consistently observe relationships between ADHD and substance use and abuse outcomes could be due to reliance on less-sensitive categorical diagnoses. PMID- 17909127 TI - Panic attacks and risk of incident cardiovascular events among postmenopausal women in the Women's Health Initiative Observational Study. AB - CONTEXT: Previous studies have documented an association of depression and phobic anxiety with cardiovascular morbidity and mortality, but little is known about the cardiovascular sequelae of panic anxiety. OBJECTIVE: To determine whether panic attacks are associated with risk of cardiovascular morbidity and mortality in postmenopausal women. DESIGN: Prospective cohort survey. SETTING: Ten clinical centers of the 40-center Women's Health Initiative. PARTICIPANTS: A total of 3369 community-dwelling, generally healthy postmenopausal women (aged 51-83 years) enrolled between 1997 and 2000 in the Myocardial Ischemia and Migraine Study who completed a questionnaire about occurrence of panic attacks in the previous 6 months. MAIN OUTCOME MEASURES: Cardiovascular/cerebrovascular outcomes (fatal and nonfatal myocardial infarction and stroke) and all-cause mortality were ascertained after a mean of 5.3 years of follow-up. RESULTS: A 6-month history of full-blown panic attacks, endorsed by 10% of postmenopausal women in this cohort, was associated with both coronary heart disease (hazard ratio, 4.20; 95% confidence interval, 1.76-9.99) and the combined end point of coronary heart disease or stroke (hazard ratio, 3.08; 95% confidence interval, 1.60-5.94) after controlling for multiple potential confounders. The hazard ratio for all-cause mortality, excluding those with a history of cardiovascular/cerebrovascular events, was 1.75 (95% confidence interval, 1.04-2.94). CONCLUSION: Panic attacks are relatively common among postmenopausal women and appear to be an independent risk factor for cardiovascular morbidity and mortality in older women. PMID- 17909129 TI - Effects of a psychosocial family-based preventive intervention on cortisol response to a social challenge in preschoolers at high risk for antisocial behavior. AB - CONTEXT: Salivary cortisol levels during social challenge relate to adaptive functioning in children and adults. Low cortisol levels have been related to conduct problems and antisocial behavior. Although studies in rodents implicate early-life social experience in cortisol regulation, no studies with humans have examined the effects of an experimentally manipulated early-life social experience on cortisol regulation. OBJECTIVE: To examine the effects of experimental manipulations of social experience on cortisol response to a social challenge in preschoolers at risk for antisocial behavior. DESIGN: Randomized controlled trial. SETTING: Department of Child and Adolescent Psychiatry, New York University School of Medicine. PARTICIPANTS: Ninety-two preschool-age siblings of youths adjudicated for delinquent acts. Intervention Family-based intervention included 22 weekly group sessions for parents and preschoolers and 10 biweekly home visits conducted during a 6- to 8-month period. MAIN OUTCOME MEASURES: Salivary cortisol levels before and after a social challenge (entry into an unfamiliar peer group). RESULTS: Relative to controls, children in the intervention condition had increased cortisol levels in anticipation of the peer social challenge. Increases were relative to both preintervention cortisol levels during the challenge and cortisol levels in the home, which were not altered by the intervention. CONCLUSIONS: A family-based preventive intervention for children at high risk for antisocial behavior alters stress response in anticipation of a peer social challenge. The experimentally induced change in cortisol levels parallels patterns found in normally developing, low-risk children. PMID- 17909128 TI - Effect of Alzheimer disease risk on brain function during self-appraisal in healthy middle-aged adults. AB - CONTEXT: Asymptomatic middle-aged adult children of patients with Alzheimer disease (AD) recently were found to exhibit functional magnetic resonance imaging (fMRI) deficits in the mesial temporal lobe during an encoding task. Whether this effect will be observed on other fMRI tasks is yet unknown. This study examines the neural substrates of self-appraisal (SA) in persons at risk for AD. Accurate appraisal of deficits is a problem for many patients with AD, and prior fMRI studies of healthy young adults indicate that brain areas vulnerable to AD such as the anterior mesial temporal lobe and posterior cingulate are involved during SA tasks. OBJECTIVE: To determine whether parental family history of AD (hereafter referred to as FH) or presence of the epsilon4 allele of the apolipoprotein E gene (APOE4) exerts independent effects on brain function during SA. DESIGN: Cross-sectional factorial design in which APOE4 status (present vs absent) was one factor and FH was the other. All participants received cognitive testing, genotyping, and an fMRI task that required subjective SA decisions regarding trait adjective words in comparison with semantic decisions about the same words. SETTING: An academic medical center with a research-dedicated 3.0-T MR imaging facility. PARTICIPANTS: Cognitively normal middle-aged adults (n = 110), 51 with an FH and 59 without an FH. MAIN OUTCOME MEASURE: Blood oxygen dependent contrast measured using T2*-weighted echo-planar imaging. RESULTS: Parental family history of AD and APOE4 status interacted in the posterior cingulate and left superior and medial frontal regions. There were main effects of FH (FH negative > FH positive) in the left hippocampus and ventral posterior cingulate. There were no main effects of APOE genotype. CONCLUSIONS: Our results suggest that FH may affect brain function during subjective SA in regions commonly affected by AD. Although the participants in this study were asymptomatic and middle-aged, the findings suggest that there may be subtle alterations in brain function attributable to AD risk factors. PMID- 17909130 TI - The impact of comorbidity of mental and physical conditions on role disability in the US adult household population. AB - CONTEXT: There is limited information that accounts for comorbidity on the impact of role disability associated with a wide range of mental and physical disorders in population-based samples. OBJECTIVE: To estimate the comparative effects of common mental and physical conditions on role disability in the general population using a novel method that accounts for comorbidity. DESIGN: Direct interviews about physical and mental conditions during the past year. SETTING: The National Comorbidity Survey Replication, a nationally representative series of face-to-face interviews. PATIENTS: A nationally representative sample of adults living in households (N = 5962 respondents, 18 years and older). MAIN OUTCOME MEASURE: Disability in major life roles was assessed with the World Health Organization Disability Assessment Schedule. Simulations that allow for complex interactions among conditions were used to estimate the conditions' effects on disability days, when respondents were completely unable to carry out their usual daily activities because of problems with mental or physical health, in the past 12 months. RESULTS: An estimated 53.4% of US adults have 1 or more of the mental or physical conditions assessed in the survey. These respondents report an average 32.1 more role-disability days in the past year than demographically matched controls, equivalent to nearly 3.6 billion days of role disability in the population. Musculoskeletal disorders and major depression had the greatest effects on disability days. Mental conditions accounted for more than half as many disability days as all physical conditions at the population level. Associations of specific conditions with disability decreased substantially after controlling for comorbidity, suggesting that prior studies, which generally did not control for comorbidity, overestimated disease-specific effects. CONCLUSION: The staggering amount of health-related disability associated with mental and physical conditions should be considered in establishing priorities for the allocation of health care and research resources. PMID- 17909131 TI - Evidence of an association between the vasopressin V1b receptor gene (AVPR1B) and childhood-onset mood disorders. AB - CONTEXT: Disturbances in stress hormones have been implicated in mood disorders, in particular in the hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis. Arginine vasopressin (AVP) plays a crucial role in modulating the HPA axis under stress and does so through a G protein-coupled receptor, vasopressin V1b receptor (AVPR1b). OBJECTIVE: To determine if genetic variation in AVPR1B could be contributing to vulnerability to mood disorders. DESIGN: We genotyped single nucleotide polymorphisms (SNPs) across the AVPR1B gene in a family-based sample with childhood-onset mood disorders. Six SNPs were genotyped; 2 were novel nonsynonymous polymorphisms, and the other 4 were constituents of a haplotype that was previously shown to be protective against depression. SETTING: Twenty three mental health facilities in Hungary. PARTICIPANTS: The sample was composed of 382 Hungarian nuclear families ascertained through affected probands with a diagnosis of childhood-onset mood disorder. MAIN OUTCOME MEASURES: Association with childhood-onset mood disorders was tested using the transmission disequilibrium test, which measures the transmission frequency of alleles, or haplotypes, from parents to affected offspring. RESULTS: Two of the AVPR1B SNPs showed association individually (Lys65Asn: chi(2) = 7.81, P = .005; S4: chi(2) = 4.58, P = .03); of particular interest is Lys65Asn, which causes an amino acid change in an intracellular protein domain. Haplotype analysis demonstrated significant overtransmission of the most frequent haplotype (chi(2)(3) = 22.42, P <.001). Furthermore, stratifying the sample by sex established that the association was predominantly in affected females, which is consistent with previous observations. CONCLUSIONS: We have found evidence to implicate the AVPR1B gene in the etiology of mood disorders, particularly in females. Antagonists of AVPR1b exhibit antidepressant qualities; hence, genetic variation in AVPR1B may have implications in HPA axis dysregulation in mood disorders. PMID- 17909132 TI - Understanding mental health treatment in persons without mental diagnoses: results from the National Comorbidity Survey Replication. AB - CONTEXT: Epidemiologic surveys have consistently found that approximately half of respondents who obtained treatment for mental or substance use disorders in the year before interview did not meet the criteria for any of the disorders assessed in the survey. Concerns have been raised that this pattern might represent evidence of misallocation of treatment resources. OBJECTIVE: To examine patterns and correlates of 12-month treatment of mental health or substance use problems among people who do not have a 12-month DSM-IV disorder. DESIGN AND SETTING: Data are from the National Comorbidity Survey Replication, a nationally representative face-to-face US household survey performed between February 5, 2001, and April 7, 2003, that assessed DSM-IV disorders using a fully structured diagnostic interview, the World Health Organization Composite International Diagnostic Interview (CIDI). PARTICIPANTS: A total of 5692 English-speaking respondents 18 years and older. MAIN OUTCOME MEASURES: Patterns of 12-month service use among respondents without any 12-month DSM-IV CIDI disorders. RESULTS: Of respondents who used 12-month services, 61.2% had a 12-month DSM-IV CIDI diagnosis, 21.1% had a lifetime but not a 12-month diagnosis, and 9.7% had some other indicator of possible need for treatment (subthreshold 12-month disorder, serious 12-month stressor, or lifetime hospitalization). The remaining 8.0% of service users accounted for only 5.6% of all services and even lower proportions of specialty (1.9%-2.4%) and general medical (3.7%) visits compared with higher proportions of human services (18.9%) and complementary and alternative medicine (7.6%) visits. Only 26.5% of the services provided to the 8.0% of presumably low-need patients were delivered in the mental health specialty or general medical sectors. CONCLUSIONS: Most services provided for emotional or substance use problems in the United States go to people with a 12-month diagnosis or other indicators of need. Patients who lack these indicators of need receive care largely outside the formal health care system. PMID- 17909133 TI - Conscientiousness and the incidence of Alzheimer disease and mild cognitive impairment. AB - CONTEXT: The personality trait of conscientiousness has been related to morbidity and mortality in old age, but its association with the development of Alzheimer disease is not known. OBJECTIVE: To test the hypothesis that a higher level of conscientiousness is associated with decreased risk of Alzheimer disease. DESIGN: Longitudinal clinicopathologic cohort study with up to 12 years of annual follow up. SETTING: The Religious Orders Study. PARTICIPANTS: A total of 997 older Catholic nuns, priests, and brothers without dementia at enrollment, recruited from more than 40 groups across the United States. At baseline, they completed a standard 12-item measure of conscientiousness. Those who died underwent a uniform neuropathologic evaluation from which previously established measures of amyloid burden, tangle density, Lewy bodies, and chronic cerebral infarction were derived. MAIN OUTCOME MEASURES: Clinical diagnosis of Alzheimer disease and change in previously established measures of global cognition and specific cognitive functions. RESULTS: Conscientiousness scores ranged from 11 to 47 (mean, 34.0; SD, 5.0). During follow-up, 176 people developed Alzheimer disease. In a proportional hazards regression model adjusted for age, sex, and education, a high conscientiousness score (90th percentile) was associated with an 89% reduction in risk of Alzheimer disease compared with a low score (10th percentile). Results were not substantially changed by controlling for other personality traits, activity patterns, vascular conditions, or other risk factors. Conscientiousness was also associated with decreased incidence of mild cognitive impairment and reduced cognitive decline. In those who died and underwent brain autopsy, conscientiousness was unrelated to neuropathologic measures, but it modified the association of neurofibrillary pathologic changes and cerebral infarction with cognition proximate to death. CONCLUSION: Level of conscientiousness is a risk factor for Alzheimer disease. PMID- 17909134 TI - Disability in children and young adults: the unintended consequences. PMID- 17909135 TI - A family-centered, community-based system of services for children and youth with special health care needs. AB - OBJECTIVE: To present a conceptual definition of a family-centered system of services for children and youth with special health care needs (CYSHCN). Previous work by the Maternal and Child Health Bureau to define CYSHCN has had widespread program effects. This article similarly seeks to provide a definition of a system of services. DESIGN: Comprehensive literature review of systems of services and consensus panel organized to review and refine the definition. SETTING: Policy research group and advisors at multiple sites. PARTICIPANTS: Policy researchers, content experts on CYSHCN, family representatives, and state program directors. OUTCOME: Definition of a system of services for CYSHCN. RESULTS: This article defines a system of services for CYSHCN as a family-centered network of community based services designed to promote the healthy development and well-being of these children and their families. The definition can guide discussion among policy makers, practitioners, state programs, researchers, and families for implementing the "community-based systems of services" contained in Title V of the Social Security Act. Critical characteristics of a system include coordination of child and family services, effective communication among providers and the family, family partnership in care provision, and flexibility. CONCLUSIONS: This definition provides a conceptual model that can help measurement development and assessment of how well systems work and achieve their goals. Currently available performance objectives for the provision of care for CYSHCN and national surveys of child health could be modified to assess systems of services in general. PMID- 17909136 TI - A tertiary care-primary care partnership model for medically complex and fragile children and youth with special health care needs. AB - OBJECTIVE: To evaluate the impact of a tertiary care center special needs program that partners with families and primary care physicians to ensure seamless inpatient and outpatient care and assist in providing medical homes. DESIGN: Up to 3 years of preenrollment and postenrollment data were compared for patients in the special needs program from July 1, 2002, through June 30, 2005. SETTING: A tertiary care center pediatric hospital and medical school serving urban and rural patients. PARTICIPANTS: A total of 227 of 230 medically complex and fragile children and youth with special needs who had a wide range of chronic disorders and were enrolled in the special needs program. INTERVENTIONS: Care coordination provided by a special needs program pediatric nurse case manager with or without a special needs program physician. MAIN OUTCOME MEASURES: Preenrollment and postenrollment tertiary care center resource utilization, charges, and payments. RESULTS: A statistically significant decrease was found in the number of hospitalizations, number of hospital days, and tertiary care center charges and payments, and an increase was found in the use of outpatient services. Aggregate data revealed a decrease in hospital days from 7926 to 3831, an increase in clinic visits from 3150 to 5420, and a decrease in tertiary care center payments of $10.7 million. The special needs program budget for fiscal year 2005 had a deficit of $400,000. CONCLUSION: This tertiary care-primary care partnership model improved health care and reduced costs with relatively modest institutional support. PMID- 17909137 TI - Impaired motor competence in school-aged children with complex congenital heart disease. AB - OBJECTIVE: To explore the extent and type of motor problems in children with complex congenital heart disease (CHD) compared with schoolchildren without any documented heart failure. DESIGN: Cohort study. SETTING: Biomechanical Laboratory, Rikshospitalet-Radiumhospitalet Medical Centre. PARTICIPANTS: One hundred twenty children aged 7 to 12 years with complex CHD and 387 healthy schoolchildren in the same age range (control group). INTERVENTIONS: All children with CHD were surgically treated with multiple corrections within the first year of life. MAIN OUTCOME MEASURES: Movement Assessment Battery for Children, grip strength, quadriceps muscle strength, and balance. RESULTS: Compared with the control group, children with CHD had a risk of having any degree of impaired motor competence of 5.8 (95% confidence interval, 3.8-8.8). The risk for having severe motor problems was 11.0 (95% confidence interval, 5.4-22.5). There were highly significant differences between the groups for manual dexterity, ball skills, grip strength, quadriceps muscle strength, and static and dynamic balance (P<.001). CONCLUSIONS: Children with CHD have a risk of severe motor problems 11 fold that of schoolchildren without any known heart failure. This suggests that primary health care providers should screen the motor competence in children with CHD at an early age to initiate therapeutic actions for children who show incipient motor problems. Optimal rehabilitative, social, and environmental support may improve the children's motor competence and prevent future health problems. PMID- 17909138 TI - Hypothermia to treat neonatal hypoxic ischemic encephalopathy: systematic review. AB - OBJECTIVES: To systematically review the effectiveness, as determined by survival without moderate to severe neurodevelopmental disability in infancy and childhood, and the safety of hypothermia vs normothermia in neonates with postintrapartum hypoxic-ischemic encephalopathy and to perform subgroup analyses based on severity of encephalopathy (moderate or severe), type of hypothermia (systemic or selective head cooling), and degree of hypothermia (moderate [or=33.6 degrees C]). DATA SOURCES: MEDLINE, EMBASE, CINAHL (Cumulative Index for Nursing and Allied Health Literature), the Cochrane Library, abstracts of annual meetings of the Pediatric Academic Societies, and bibliographies of identified articles. STUDY SELECTION: Randomized and quasi-randomized controlled trials without language restriction were assessed by 2 reviewers independently and discrepancies were resolved by involving a third reviewer. Quality of the trials was assessed on the basis of concealment of allocation, method of randomization, masking of outcome assessment, and completeness of follow-up. INTERVENTION: Systemic or selective head hypothermia compared with normothermia. MAIN OUTCOME MEASURE: Death or moderate to severe neurodevelopmental disability. RESULTS: Eight studies of acceptable quality were included. The combined outcome of death or neurodevelopmental disability in childhood was reduced in infants receiving hypothermia compared with control infants (4 studies including 497 infants; relative risk, 0.76, 95% confidence interval, 0.65-0.88; number needed to treat, 6; 95% confidence interval, 4-14), as were death and moderate to severe neurodevelopmental disability when analyzed separately. Cardiac arrhythmias and thrombocytopenia were more common with hypothermia; however, they were clinically benign. CONCLUSIONS: In neonates with postintrapartum asphyxial hypoxic-ischemic encephalopathy, hypothermia is effective in reducing death and moderate to severe neurodevelopmental disability either in combination or separately and is a safe intervention. PMID- 17909139 TI - Predictors of initiation of alcohol use among US adolescents: findings from a prospective cohort study. AB - OBJECTIVE: To identify precursors of adolescent alcohol initiation and binge drinking. DESIGN: Prospective cohort study. SETTING: Self-report questionnaires. PARTICIPANTS: A total of 5511 Growing Up Today Study participants aged 11 to 18 years in 1998. Main Exposures Individual, family, and social factors. MAIN OUTCOME MEASURES: First whole drink of alcohol and binge drinking. RESULTS: Between 1998 and 1999, 611 girls (19%) and 384 boys (17%) initiated alcohol use. Older age, later maturational stage, smoking, adults drinking in the home, underage sibling drinking, peer drinking, possession of or willingness to use alcohol promotional items, and positive attitudes toward alcohol were associated with an increased likelihood of alcohol initiation. Girls who ate family dinner at home every day were less likely to initiate alcohol use than girls who ate family dinner only on some days or never (odds ratio, 0.66; 95% confidence interval, 0.50-0.87). Girls with higher social self-esteem and boys with higher athletic self-esteem were more likely to initiate alcohol use than those with lower self-esteem. Among teens who initiated alcohol use, 149 girls (24%) and 112 boys (29%) further engaged in binge drinking. Among girls, positive attitudes toward alcohol, underage sibling drinking, and possession of or willingness to use alcohol promotional items were associated with binge drinking; among boys, positive attitudes toward alcohol and older age were associated with binge drinking. CONCLUSIONS: Eating family dinner at home every day may delay alcohol uptake among some adolescents. Alcohol promotional items appear to encourage underage alcohol initiation and binge drinking; this may warrant marketing restrictions on the alcohol industry. PMID- 17909140 TI - Effect of block play on language acquisition and attention in toddlers: a pilot randomized controlled trial. AB - OBJECTIVE: To test the hypotheses that block play improves language acquisition and attention. DESIGN: Randomized controlled trial. SETTING: Pediatric clinic. PARTICIPANTS: Children aged 1(1/2) to 2(1/2) years. INTERVENTION: Distribution of 2 sets of building blocks. MAIN OUTCOME MEASURES: Scores on the MacArthur-Bates Communicative Development Inventories, television viewing based on diary data, and the hyperactivity domain of the Child Behavior Checklist. RESULTS: Of 220 families approached in the clinic waiting room, 175 (80%) agreed to participate in the study. At least 1 diary was returned from 92 of the 175 families (53%). A total of 140 families (80%) completed exit interviews. Of the children in the intervention group, 52 (59%) had block play reported in their diaries compared with 11 (13%) in the control group (P<.01). The linear regression results for language acquisition were as follows: entire sample--raw score, 7.52 (P=.07); percentile, 8.4 (P=.15); low-income sample--raw score, 12.40 (P=.01); percentile, 14.94 (P=.03). For attention the results were as follows: entire sample--odds ratio, 0.49 (P=.29); low-income sample--odds ratio, 0.48 (P=.26) There were no statistically significant differences with respect to hyperactivity scores. CONCLUSIONS: Distribution of blocks can lead to improved language development in middle- and low-income children. Further research is warranted. PMID- 17909141 TI - Infections in child day care centers and later development of asthma, allergic rhinitis, and atopic dermatitis: prospective follow-up survey 12 years after controlled randomized hygiene intervention. AB - OBJECTIVE: To evaluate the effect of successful prevention of common infections in child day care centers on the later development of allergic diseases. DESIGN: Prospective follow-up survey with a questionnaire administered 12 years after a controlled randomized hygiene intervention. SETTING: Twenty municipal child day care centers in Oulu, Finland. PARTICIPANTS: A questionnaire was sent to 1354 prior participants (98%) in the intervention trial. The response rate was 68% (928 of 1354 participants). MAIN INTERVENTION: Hygiene intervention from March 1, 1991, to May 31, 1992. MAIN OUTCOME MEASURES: The number of respondents who had a diagnosis of asthma, allergic rhinitis, and/or atopic dermatitis made by a physician, and the number of those who reported symptoms of atopic diseases. RESULTS: Asthma was diagnosed by a physician in 48 of the 481 respondents (10%) from the intervention child day care centers, with markedly fewer infections, and in 46 of the 447 controls (10%) (relative risk, 1.0; 95% confidence interval, 0.7 1.4). Similarly, no differences were found in the numbers of children who had a diagnosis of other atopic diseases or who had reported such symptoms. CONCLUSION: The prevention of common respiratory tract and enteric infections during early childhood does not change later allergic morbidity. PMID- 17909142 TI - Dietary supplement use among infants, children, and adolescents in the United States, 1999-2002. AB - OBJECTIVE: To describe dietary supplement use among US children. DESIGN: Analysis of nationally representative data from the 1999-2002 National Health and Nutrition Examination Survey (NHANES). SETTING: Home interviews and a mobile examination center. PARTICIPANTS: Children from birth through 18 years who participated in NHANES (N=10,136). MAIN EXPOSURE: Frequency of use of any dietary supplement product. OUTCOME MEASURE: Prevalence of use and intake of key nutrients from supplements among children. RESULTS: In 1999-2002, 31.8% of children used dietary supplements, with the lowest use reported among infants younger than 1 year (11.9%) and teenagers 14 to 18 years old (25.7%) and highest use among 4- to 8-year-old children (48.5%). Use was highest among non-Hispanic white (38.1%) and Mexican American (22.4%) participants, lowest among non Hispanic black participants (18.8%), and was not found to differ by sex. The type of supplement most commonly used was multivitamins and multiminerals (18.3%). Ascorbic acid (28.6%), retinol (25.8%), vitamin D (25.6%), calcium (21.1%), and iron (19.3%) were the primary supplemental nutrients consumed. Supplement use was associated with families with higher incomes; a smoke-free environment; not being certified by the US Department of Agriculture Special Supplemental Nutrition Program for Women, Infants and Children in the last 12 months; lower child body mass index; and less daily recreational screen time (television, video games, computers, etc) (P<.005). The highest prevalence of supplement use (P<.005) was in children who were underweight or at risk for underweight (P<.005). CONCLUSIONS: More than 30% of children in the United States take dietary supplements regularly, most often multivitamins and multiminerals. Given such extensive use, nutrient intakes from dietary supplements must be included to obtain accurate estimates of overall nutrient intake in children. PMID- 17909143 TI - Homelessness and health care access after emancipation: results from the Midwest Evaluation of Adult Functioning of Former Foster Youth. AB - OBJECTIVE: To estimate the association between housing status and health care access and outcomes among young adults aging out of the child welfare system. DESIGN: Prospective cohort study SETTING: Illinois, Iowa, and Wisconsin. Baseline interviews were conducted between May 2002 and March 2003 and follow-up interviews, between March and December 2004. PARTICIPANTS: Participants were foster youth aged 17 or 18 years in Illinois, Wisconsin, or Iowa. We invited a random sample of 67% of eligible Illinois youth and all eligible youth from Wisconsin and Iowa to participate. Researchers interviewed 749 at baseline (94.7% response) and 643 at follow-up (85.8%); we excluded 8 participants without housing data (n=635). We included only the 345 emancipated participants in analyses of health care access. MAIN EXPOSURE Housing status after emancipation: stable housing; unstable housing; or homeless. MAIN OUTCOME MEASURES: Multivariate adjusted odds ratio (AOR) of association between main exposure variables with 3 measures of access to care and 2 health outcomes. RESULTS: Among the 345 emancipated participants, 14.2% experienced homelessness and 39.4% were unstably housed. In multivariate analysis of emancipated participants, homelessness was associated with being uninsured (AOR, 3.41; 95% confidence interval, 1.52-7.63) and having unmet need for health care (AOR, 3.26; 95% confidence interval, 1.40-7.56); it was not associated with not having had ambulatory care. In multivariate analysis of all participants, housing status was not associated with reporting fair or poor health at follow-up or, among women, with having had a pregnancy. CONCLUSION: Having had an episode of homelessness after emancipation is associated with worse health access, but not worse outcomes, among youth emancipated from foster care. PMID- 17909144 TI - Long-term follow-up data collection and use in state newborn screening programs. AB - OBJECTIVES: To describe and analyze the types of data-related policies and practices that currently exist among state newborn screening (NBS) programs in relation to long-term follow-up (LTFU) and oversight for newborns with confirmed disorders. DESIGN: A 19-question online survey. PARTICIPANTS: Thirty-five state NBS programs. MAIN OUTCOME MEASURES: Whether LTFU is performed, collection and use of LTFU data, and variety of LTFU data collected. RESULTS: Survey findings reveal data-related challenges faced by state NBS programs in their ability to perform ongoing oversight, evaluation, and quality assurance with respect to LTFU for newborns with confirmed disorders. Of the NBS programs surveyed, 56% reported collecting no LTFU data. More than two-thirds of state NBS programs surveyed do not use LTFU data at all or use it only minimally. Most programs that collect any LTFU data from providers (physicians, nurses, and allied health professionals) do it through verbal communication or paper forms rather than electronically. Almost half of the programs collecting any LTFU data do so only once a year. A lot of variety exists in the types of LTFU data collected across programs. Most of the 15 programs that reported collecting LTFU data use it to track the clinical outcomes of patients, assess the needs of patients and their families for services, and track and identify individuals lost to follow-up across time. CONCLUSION: The results generally point to a need for greater alignment of state NBS program data practices and policies with the data requirements for essential public health functions, such as quality assurance, program evaluation, and cost benefit analysis. PMID- 17909145 TI - Picture of the month. Diagnosis: periocular hemangioma: an eye-opening experience. PMID- 17909146 TI - Being specific about being special: defining children's conditions and special health care needs. PMID- 17909147 TI - Providing better opportunities for older children in the child welfare system. PMID- 17909148 TI - A need for pediatric genetics. PMID- 17909150 TI - Neutralizing antibodies to biological therapies: A "touch of gray" vs a "black and white" story. PMID- 17909151 TI - The plate is overflowing and it's not enough: binge eating in frontotemporal lobar degeneration. PMID- 17909152 TI - Minocycline treatment in acute stroke: an open-label, evaluator-blinded study. AB - BACKGROUND: Ischemic animal model studies have shown a neuroprotective effect of minocycline. OBJECTIVE: To analyze the effect of minocycline treatment in human acute ischemic stroke. METHODS: We performed an open-label, evaluator-blinded study. Minocycline at a dosage of 200 mg was administered orally for 5 days. The therapeutic window of time was 6 to 24 hours after onset of stroke. Data from NIH Stroke Scale (NIHSS), modified Rankin Scale (mRS), and Barthel Index (BI) were evaluated. The primary objective was to compare changes from baseline to day 90 in NIHSS in the minocycline group vs placebo. RESULTS: One hundred fifty-two patients were included in the study. Seventy-four patients received minocycline treatment, and 77 received placebo. NIHSS and mRS were significantly lower and BI scores were significantly higher in minocycline-treated patients. This pattern was already apparent on day 7 and day 30 of follow-up. Deaths, myocardial infarctions, recurrent strokes, and hemorrhagic transformations during follow-up did not differ by treatment group. CONCLUSIONS: Patients with acute stroke had significantly better outcome with minocycline treatment compared with placebo. The findings suggest a potential benefit of minocycline in acute ischemic stroke. PMID- 17909153 TI - Cognitive impairment in familial ALS. AB - BACKGROUND: ALS is a progressive neurodegenerative disorder that affects upper and lower motor neurons. Recent reports demonstrate cognitive impairment in patients with sporadic ALS (sALS). OBJECTIVE: To test whether patients with familial ALS (fALS) have cognitive impairment and whether it is of the same type and degree as observed in sporadic ALS. METHODS: Thirty-seven consecutive patients with fALS underwent comprehensive neuropsychological testing. Cognitive impairment was categorized by 1) cluster analysis of non-timed, non-motor dependent neuropsychological tests, 2) cutoff scores, and 3) clinical impression. RESULTS: By cluster analysis, 23 of 37 (62%) patients with fALS and 190 of 392 (48.5%) patients with sALS had cognitive impairment (difference not significant). Similar rates of impairment were found using clinical diagnostic criteria and cutoff score analysis. Neither motor scores nor the site of symptom onset correlated with cognitive impairment. Only age differed between the affected and unaffected fALS groups. CONCLUSIONS: The prevalence and pattern of cognitive impairment in familial ALS (fALS) is similar to that of sporadic ALS. For patients with fALS, the site of symptom onset did not correlate with cognitive impairment, but age did. Future studies will determine whether cognitive impairment in fALS correlates with specific genetic mutations or polymorphisms. PMID- 17909154 TI - Serum uric acid and brain ischemia in normal elderly adults. AB - BACKGROUND: Uric acid (UA) has antioxidant properties yet when elevated is associated with vascular disease and stroke. Further, even high normal UA is associated with increased risk of mild cognitive dysfunction in elderly adults. METHOD: In this cross-sectional, observational study, we examined the relationship between serum UA and aggregate volume of white matter hyperintense (WMH) signals observed on proton density and T2-weighted brain MR images in a community sample of 177 adults ages 20 to 92. Using logistic regression, we tested whether participants with UA concentrations in the highest quartile of the sample--but still normal--would have increased WMH volumes. RESULTS: Compared with those with low to moderate levels, participants with high normal serum UA were more likely to fall in the highest quartile of WMH volume. The odds ratios (95% CIs) of increased WMH were 2.6 (1.2 to 5.4) for total, 2.5 (1.2 to 5.1) for periventricular, and 2.8 (1.4 to 5.9) for subcortical WMH volume. After controlling for age, sex, race, education, body mass, hypertension, and diabetes, the multivariate-adjusted odds of large total and subcortical WMH volumes remained elevated. Finally, high normal UA increased the odds of having excessive ischemic burden four- to fivefold in adults ages 60 and older. CONCLUSIONS: These findings demonstrate that mildly elevated serum uric acid is associated with increased burden of cerebral ischemic pathology, particularly in older adults. We outline the potential pathogenesis of this association. A clinical trial of antihyperuricemic medication to treat or prevent chronic brain ischemia might be warranted. PMID- 17909155 TI - Binge eating is associated with right orbitofrontal-insular-striatal atrophy in frontotemporal dementia. AB - BACKGROUND: Neurophysiologic studies on human and nonhuman primates implicate an orbitofrontal-insular-striatal circuit in high-level regulation of feeding. However, the role of these areas in determining feeding disturbances in neurologic patients remains uncertain. OBJECTIVE AND METHODS: To determine brain structures critical for control of eating behavior, we performed a prospective, laboratory-based, free-feeding study of 18 healthy control subjects and 32 patients with neurodegenerative disease. MR voxel-based morphometry (VBM) was used to identify regions of significant atrophy in patients who overate compared with those who did not. RESULTS: Despite normal taste recognition, 6 of 32 patients compulsively binged, consuming large quantities of food after reporting appropriate satiety. All six patients who overate were clinically diagnosed with frontotemporal dementia (FTD), a disorder previously associated with disordered eating, while the nonovereaters were diagnosed with FTD, semantic dementia, progressive aphasia, progressive supranuclear palsy, and Alzheimer disease. VBM revealed that binge-eating patients had significantly greater atrophy in the right ventral insula, striatum, and orbitofrontal cortex. CONCLUSION: Binge eating can occur despite reported satiety and is associated with damage to a right-sided orbitofrontal-insular-striatal circuit in humans. These findings support a model in which ventral insular and orbitofrontal cortices serve as higher-order gustatory regions and cooperate with the striatum to guide appropriate feeding responses. PMID- 17909157 TI - Education and dementia: what lies behind the association? AB - BACKGROUND: Low education seems to be associated with an increased risk of dementia and Alzheimer disease (AD). People with low education have unhealthier lifestyles and more cardiovascular risk factors, but it is unclear how this affects the association between education and dementia. METHODS: Participants of the Cardiovascular Risk Factors, Aging and Dementia (CAIDE) study were derived from random, population-based samples previously studied in a survey in 1972, 1977, 1982, or 1987. After an average follow-up of 21 years, 1,449 individuals (72%) aged 65 to 79 participated in a re-examination in 1998. RESULTS: Compared to individuals with formal education of 5 years or less, those with 6 to 8 years of education had OR of 0.57 (95% CI 0.29 to 1.13), and those with 9 years of education or more had OR of 0.16 (95% CI 0.06 to 0.41) for dementia. The corresponding ORs for AD were 0.49 (0.24 to 1.00) and 0.15 (0.05 to 0.40). The associations remained unchanged after adjustments for several demographic, socioeconomic, vascular, and lifestyle characteristics. The results were similar among both men and women. ApoE4 did not modify the association, but the risk of dementia and AD was very low among ApoE4 noncarriers with high education. CONCLUSIONS: The association between low education and dementia is probably not explained by the unhealthy lifestyles of the less educated compared with higher educated persons. Higher educated persons may have a greater cognitive reserve that can postpone the clinical manifestation of dementia. Unhealthy lifestyles may independently contribute to the depletion of this reserve or directly influence the underlying pathologic processes. PMID- 17909156 TI - Predictors of fitness to drive in people with Parkinson disease. AB - OBJECTIVE: To develop an efficient clinical screening battery to accurately predict the fitness to drive in people with Parkinson disease (PD). METHODS: This prospective study included 80 participants: 40 patients with PD and 40 healthy age- and sex-matched control subjects. All participants were assessed using a driving simulator, a driving history survey, and the Clinical Dementia Rating. The patients with PD also underwent a clinical test battery and an evaluation of fitness to drive performed by an official center, which included visual, cognitive, and on-road tests. A two-class decision from this driving assessment center was the main outcome measure. RESULTS: A screening battery assessing four clinical variables (disease duration, contrast sensitivity, Clinical Dementia Rating, and motor part of the Unified Parkinson's Disease Rating Scale) provided the best model (R(2) = 0.52) to predict the fitness to drive and correctly classified 36 (90%) of the patients with PD as pass or fail (sensitivity = 91%, specificity = 90%). The Test Ride for Investigating Practical fitness to drive (TRIP) driving simulator score discriminated significantly between drivers with PD and their healthy peers (p = 0.0008). When the TRIP driving simulator score was added to the clinical model, the total explained variance increased (R(2) = 0.60) and correctly classified 39 (97.5%) of drivers with PD into the pass/fail category (sensitivity = 91%, specificity = 100%). CONCLUSIONS: A short clinical screening battery that measures disease duration, contrast sensitivity, cognitive and motor functions can predict fitness to drive in people with Parkinson disease with a high degree of accuracy. PMID- 17909158 TI - Differences and similarities between atypical facial pain and trigeminal neuropathic pain. AB - OBJECTIVE: To investigate contribution of neuropathic mechanisms to clinically diagnosed atypical facial pain (AFP) using neurophysiologic and thermal quantitative sensory testing (QST) and comparing findings in AFP with those in definite trigeminal neuropathic pain (TNP). METHODS: Twenty patients with AFP and 12 patients with TNP participated after thorough clinical diagnostic workup. All patients underwent blink reflex (BR) recordings, habituation of the BR, and (except one patient with TNP) thermal QST. The results were compared with the reference values of our laboratory for normality. RESULTS: Of the patients with AFP, 75% showed abnormal findings. The BR responses were abnormal in three (15%) AFP patients (in two patients, the findings were compatible with a peripheral neuropathy and in one with a brainstem lesion), and in seven (58%) TNP patients. Seven (35%) patients with AFP and four (33%) with TNP showed increased excitability of the BR in the form of deficient habituation. Thermal QST indicated abnormal small fiber function in 11 (55%) patients with AFP and in all patients with TNP tested. QST showed thermal hypoesthesia in 45% and warm allodynia in 10% of patients with AFP. In TNP, all findings indicated thermal hypoesthesia. Abnormalities in BR and thermal QST were less frequent in AFP than TNP, but when present, type and pattern of findings were similar in both conditions. CONCLUSIONS: Clinical diagnosis of atypical facial pain represents a heterogeneous entity and seems to form a continuum regarding the level and extent of neuropathic involvement. Without detailed neurophysiologic and quantitative sensory examinations, neuropathic cause of chronic orofacial pain may be overlooked. PMID- 17909159 TI - Reduced cardiac 123I-MIBG uptake reflects cardiac sympathetic dysfunction in Lewy body disease. AB - OBJECTIVE: To examine the relation between the results of cardiac (123)I-meta iodobenzylguanidine (MIBG) scintigraphy and cardiovascular autonomic function in Lewy body disease (LBD). METHODS: The subjects were 66 patients with LBD, 44 of whom had Parkinson disease (PD), 10 PD with dementia (PDD), and 12 dementia with Lewy bodies (DLB); 20 age-matched healthy subjects were studied as controls. Cardiovascular autonomic function was evaluated on the basis of cardiac (123)I MIBG uptake, cardiovascular autonomic response on the Valsalva maneuver (VM), and systolic blood pressure (SBP) response on head-up tilt table (HUT) testing. RESULTS: Patients with LBD had reduced cardiac (123)I-MIBG uptake, cardiovascular autonomic response on the VM, and SBP response on HUT testing as compared with controls. Cardiac (123)I-MIBG uptake and cardiovascular autonomic function in PDD and DLB were severely impaired as compared with those in PD. Cardiac (123)I-MIBG uptake in LDB was not significantly related to vasomotor sympathetic function, baroreceptor reflex gain, cardiac parasympathetic function, or the changes in SBP on HUT testing. Cardiac (123)I-MIBG uptake was, however, significantly related to the blood pressure overshoot in phase IV of the VM. CONCLUSION: Cardiac (123)I meta-iodobenzylguanidine uptake clinically reflects cardiac sympathetic dysfunction in Lewy body disease. PMID- 17909160 TI - Sign language aphasia due to left occipital lesion in a deaf signer. AB - Localization of sign language production and comprehension in deaf people has been described as similar to that of spoken language aphasia. However, sign language employs a visuospatial modality through visual information. We present the first report of a deaf signer who showed substantial sign language aphasia with severe impairment in word production due to a left occipital lesion. This case may indicate the possibility of other localizations of plasticity. PMID- 17909161 TI - Re: Low LDL cholesterol, statins, and brain hemorrhage: should we worry? PMID- 17909162 TI - Tissue factor: a critical role in inflammation and cancer. AB - A series of coordinated enzymatic reactions takes place in the body whenever blood clots. The major physiological initiator of these reactions is a membrane bound glycoprotein known as tissue factor (TF), which is normally separated from the bloodstream by the vascular endothelium. Bleeding, caused by injury or tissue damage, activates a complex enzyme cascade as TF becomes exposed to the bloodstream. In disease states, leukocytes or the vascular endothelium may abnormally express TF to cause intravascular coagulation. The blood-coagulation cascade is also relevant to diseases such as hemophilia, in which patients are deficient in blood proteins necessary for clotting, and is linked to vascular diseases such as heart attack and stroke, in which clotting can lead to the occlusion of blood vessels. Coagulation is also activated in inflammation and cancer. In this article, we discuss characteristics of TF and review its role in inflammation and cancer. PMID- 17909163 TI - C-reactive protein in nipple aspirate fluid associated with Gail model factors. AB - BACKGROUND: The majority of breast cancers originate in the epithelial lining of the breast ductal system. Premalignant cell damage in this lining may produce biochemical signals that deliver inflammatory proteins to the site. The presence of C-reactive protein (CRP) in nipple aspirate fluid (NAF) may reflect an inflammatory state indicative of a premalignant breast microenvironment. This study ascertained CRP's presence in NAF and evaluated if risk factors, as identified by the Gail model, were associated with NAF CRP levels among healthy women. DESIGN: NAF CRP levels were assayed in 59 women. RESULTS: CRP was present in NAF and significantly (p = .04) and positively related to breast cancer risk as predicted by the Gail model. CONCLUSION: CRP is differentially present in NAF and varies by Gail model risk factors. CRP in NAF holds promise as a noninvasive biomarker that detects a precarcinogenic breast ductal microenvironment and may contribute to the diagnosis of breast cancer early in the course of the disease when prognosis is most favorable. PMID- 17909164 TI - The effects of vitamin D deficiency and insufficiency on the endocrine and paracrine systems. AB - Individuals are capable of producing vitamin D with proper exposure to sunlight. However, several factors can interfere with the effectiveness of this process. Most sunscreens filter out UVB light, thus inhibiting vitamin D production. Individuals with more darkly pigmented skin have greater difficulty producing vitamin D because melanin acts as an effective natural sunscreen, requiring longer sun exposure to produce an adequate daily allotment of vitamin D. Additionally, solely breastfed infants whose mothers suffered from vitamin D deficiency or insufficiency when pregnant have smaller reserves of the nutrient and are at greater risk of developing nutritional rickets. Vitamin D deficiency leads to rickets, osteomalacia, and osteoporosis. Long-term vitamin D insufficiency can lead to paracrine effects such as type 1 diabetes, cancer, and multiple sclerosis. This article reviews the current literature on vitamin D deficiency and insufficiency and their relation to different disease states. Potential areas for research are discussed. PMID- 17909165 TI - A feasibility study of a culturally tailored diabetes intervention for Mexican Americans. AB - Latinos, the fastest growing minority group in the United States, are among the hardest hit by diabetes. Among Latinos, Mexican Americans have the highest rate (23.9%) of diabetes. Good self-management can improve glycemic control and decrease diabetes complications but can be challenging to achieve. The purpose of this study was to test the feasibility and examine the effects of a culturally tailored intervention for Mexican Americans with type 2 diabetes on outcomes of self-management. The study used a pretest/posttest control group design with 10 participants in each group (N = 17). Feasibility and acceptability of the tailored diabetes self-management program was assessed by examining ease of recruitment and retention rates. The behavioral outcomes of self-efficacy, diabetes knowledge and self-care measures, and the biologic outcomes of weight, body mass index, HbA1C, and blood glucose were used to examine intervention effectiveness. Successful recruitment of participants came from personal referrals from providers or the promotora. Retention rates were 100% for the intervention group and 80% for the control group. Findings suggest that the intervention had a positive clinical and statistical effect on diabetes knowledge, weight, and body mass index. Improvements were also noted in self efficacy scores, blood glucose, and HbA1C, but these changes did not reach statistical significance. A culturally tailored diabetes self-management program may result in improved outcomes for Mexican Americans with type 2 diabetes. PMID- 17909166 TI - Use of general clinical research centers for nursing research. AB - This study examines to what extent academic nursing faculty members understand the purpose of general clinic research centers (GCRCs) and use this resource in their programs of research. GCRCs provide a controlled research-oriented infrastructure to conduct safe, innovative, and multidisciplinary studies. Survey questionnaires were sent to associate deans of research at schools of nursing accredited by the Commission on Collegiate Nursing Education or the National League for Nursing Accrediting Commission within 60 miles of a GCRC. They were asked to distribute questionnaires to 20% of faculty members. Of the 186 nurse faculty members responding, 85% had not conducted research at a GCRC as a principal investigator and 69% of the sample reported that their colleges or universities do not make information about GCRCs available to new faculty. Making greater use of this valuable resource allows nurse researchers to become more involved in developing new knowledge and testing interventions and in psychological and physiologic measurements, thus doing more to fulfill the mission of nursing research and increasing the involvement of nurses in the broader community of health science research. PMID- 17909167 TI - The effects of estradiol on central serotonergic systems and its relationship to mood in women. AB - Lifetime prevalence rates of depression are higher in women than men. Because this gender disparity appears after the onset of puberty and declines after menopause, gonadal hormones may play a role in women's increased vulnerability to dysphoric states. Estrogens have powerful effects beyond their role in reproduction. Fluctuations in estrogen occur naturally throughout the reproductive years and can be associated with disruptions in mood. Treatment for depression with exogenous estrogen has produced equivocal results. To shed light on the complex interactions among estrogens, serotonin, and mood, we briefly examine (a) central serotonin systems and their relationship to mood and mood disorders, (b) nonreproductive effects of estrogens on those systems, (c) potential points of intersection between serotonin systems and estrogens, and (d) research into the use of exogenous estrogen in depression in women. In conclusion, we reiterate the call for carefully controlled research into the etiology and treatment of depression in women. PMID- 17909168 TI - Relationship of SERT polymorphisms to depressive and anxiety symptoms in irritable bowel syndrome. AB - This study investigates the association of psychological symptoms with the distribution of two serotonin transporter gene (SERT) polymorphisms, located in the promoter region (5-HTTLPR) and in intron 2 (STin2 VNTR), in patients with irritable bowel syndrome (IBS). Participants, 21 men and 117 women, were assessed for mental health history and current psychological distress. A blood sample was used for genotyping. Participants who were homozygous for the short allele of 5 HTTLPR or carried a STin2.9 VNTR allele were significantly more likely to have a history of depression. Participants did not differ by genotype in their history of anxiety or suicidal ideation nor in their current levels of depression, anxiety, or general psychological distress. The results support a biopsychosocial model of IBS in which SERT genotype modifies the risk for depressive episodes. Long term, practitioners may individualize treatment of patients with IBS using genotype as one of the factors. PMID- 17909169 TI - Direct sequencing is more accurate and feasible in detecting single nucleotide polymorphisms than RFLP: using human vascular endothelial growth factor gene as a model. AB - Since the sequencing of the human genome, there has been increased interest in understanding the distribution and effects of genetic variations among individuals. Restriction fragment length polymorphism (RFLP) is a well established and frequently used method for genotyping. This method, however, is indirect and has a number of limitations. It is thus important to reevaluate the use of RFLP in light of more contemporary methods of genotyping. The specific aims of this study are to (a) compare genotyping methods of traditional RFLP with contemporary direct sequencing for accurate identification of polymorphisms within the human vascular endothelial growth factor (VEGF) gene and (b) describe distribution of a known single nucleotide polymorphism (SNP) in the VEGF gene in a sample composed of 50 healthy volunteers. Polymerase chain reaction (PCR) was used to amplify the initial sample of DNA. Genotypes of a G-to-A substitution (GG, AG, AA) at -1154 were analyzed by RFLP and direct sequencing. RFLP was unable to discriminate among the three possible genotypes, whereas direct sequencing clearly identified genotype for all 50 samples. Observed genotype frequencies were comparable with the Hardy-Weinberg principle. This comparative study provides justification for selecting direct sequencing instead of RFLP for detecting SNPs in selected genes. PMID- 17909170 TI - The clinical and economic potential of hyperbaric oxygen therapy in the treatment of diabetic ulceration and other conditions. AB - Selective use of systemic hyperbaric oxygen therapy (HBOT) is a treatment that deserves further study and analysis. The current situation in the United Kingdom (UK) is discussed in relation to available evidence and practices elsewhere. It would appear that there is increasing evidence that HBOT could benefit many patients and health care budgets through improved clinical efficacy and cost efficiency in the treatment of specific conditions, notably nonhealing diabetic ulceration of the lower limbs. This is not only disabling, it may lead to amputation. It is also a financial burden to patients and health service providers. In the UK, it is estimated that chronic wound care costs more than pound1 billion a year, with diabetic ulceration accounting for a substantial part of that staggering sum. It has been said repeatedly, and quite correctly, that there is insufficient good-quality evidence upon which a properly informed decision may be made on the contribution HBOT might make to alleviate that situation. It is intriguing that no determined effort is apparent that would seek to settle the issue by encouraging and facilitating appropriately designed and conducted randomized controlled trials to assess the actual effects of this treatment. Indeed, a proposed government research initiative appears to have been cancelled. Is lack of research preventing provision of HBOT? PMID- 17909171 TI - ECHM-ETRS joint conference on oxygen and tissue repair, Ravenna, Italy, October 27-28, 2006: recommendations by the international jury. PMID- 17909172 TI - A case report of the treatment of diabetic foot ulcers using a sodium hyaluronate and iodine complex. AB - Diabetic foot ulcers are difficult to heal due to defects in local microvasculature and persistent, concomitant infection. Despite the best medical care, amputation is often a management option for this problem. The authors have developed a new and unique system for wound treatment, which is based on a combination of high molecular weight sodium hyaluronate with an iodine complex Hyiodine (Contipro C, Dolni Dobrouc, Czech Republic). In this case report, the authors present an observational study on a series of patients with diabetic foot disease with nonhealing wounds treated with Hyiodine. The effect of the HA-iodine complex was studied on 18 patients suffering from complicated foot diabetic wounds. The HA-iodine complex was either spread directly over the wound, or more frequently, gauze was immersed in the HA-iodine complex and then put on/into the wound. Then several layers of dry gauze covered the wound. This dressing was changed every 24 hours. Wound healing was monitored daily, and wound pictures were taken each second week. Clinical improvement was observed in the majority. This suggests that the HA-iodine complex dressing has potential that needs to be developed from controlled studies. PMID- 17909173 TI - Painful ANA-positive scleroderma-like disease with acral ulcerations: a case of chronic gangrenous ergotism. AB - Chronic ergotism is a rare cause of limb ischemia. In this case report, the authors present a 62-year-old woman with history of long-term use of ergotamine alkaloids for the treatment of menstrual pain, who developed a severe painful disease initially misdiagnosed as systemic sclerosis (scleroderma) for 3 decades. She presented with a combination of acral gangrene, foot ulcer, renal obstruction, mild pulmonary fibrosis, and reduced esophageal motility. Right sided renal obstruction was evident. The condition was extremely painful and had led to muscular contractions and immobility, drug abuse, and anemia. After establishing the diagnosis of chronic gangrenous ergotism, changing drug therapy, mobilization, and treatment of chronic wounds, she showed a remarkable recovery. Eventually the foot ulcer was closed successfully using a mesh graft transplantation, and the patient was able to walk alone. Chronic ergotism is rare but has to be taken into account when presented with painful chronic digital and foot ulcers. PMID- 17909174 TI - A retained sponge is a complication of vacuum-assisted closure therapy. AB - Vacuum-assisted closure (VAC) system has become a widely used, efficacious, and overall safe method for managing many types of open wounds. The authors present a case of a rare but avoidable complication caused by a retained piece of sponge after VAC therapy (KCI Inc, San Antonio, TX). They emphasize the need for awareness and careful evaluation of the patient who receives VAC therapy to avoid diagnostic confusion and morbidity to the patient. PMID- 17909178 TI - The efficacy of honey in the treatment of wounds. PMID- 17909179 TI - Notch signaling in vascular smooth muscle cells is required to pattern the cerebral vasculature. AB - Stroke is the third leading cause of death and a significant contributor of morbidity in the United States. In humans, suboptimal cerebral collateral circulation within the circle of Willis (CW) predisposes to ischemia and stroke risk in the setting of occlusive carotid artery disease. Unique genes or developmental pathways responsible for proper CW formation are unknown. Herein we characterize a mouse model lacking Notch signaling in vascular smooth muscle cells (vSMCs), in which the animals are intolerant to reduced cerebral blood flow. Remarkably, unilateral carotid artery ligation results in profound neurological sequelae and death. After carotid ligation, perfusion of the ipsilateral cerebral hemisphere was markedly diminished, suggesting an anastomotic deficiency within the CW. High-resolution microcomputed tomographic (micro-CT) imaging revealed profound defects in cerebrovascular patterning, including interruption of the CW and anatomic deformity of the cerebral arteries. These data identify a vSMC-autonomous function for Notch signaling in patterning and collateral formation within the cerebral arterial circulation. The data further implicate genetic or functional deficiencies in Notch signaling in the pathogenesis of anatomic derangements underlying cerebrovascular accidents. PMID- 17909181 TI - Category-specific attention for animals reflects ancestral priorities, not expertise. AB - Visual attention mechanisms are known to select information to process based on current goals, personal relevance, and lower-level features. Here we present evidence that human visual attention also includes a high-level category specialized system that monitors animals in an ongoing manner. Exposed to alternations between complex natural scenes and duplicates with a single change (a change-detection paradigm), subjects are substantially faster and more accurate at detecting changes in animals relative to changes in all tested categories of inanimate objects, even vehicles, which they have been trained for years to monitor for sudden life-or-death changes in trajectory. This animate monitoring bias could not be accounted for by differences in lower-level visual characteristics, how interesting the target objects were, experience, or expertise, implicating mechanisms that evolved to direct attention differentially to objects by virtue of their membership in ancestrally important categories, regardless of their current utility. PMID- 17909180 TI - Chymotryptic specificity determinants in the 1.0 A structure of the zinc inhibited human tissue kallikrein 7. AB - hK7 or human stratum corneum chymotryptic enzyme belongs to the human tissue kallikrein (hKs) serine proteinase family and is strongly expressed in the upper layers of the epidermis. It participates in skin desquamation but is also implicated in diverse skin diseases and is a potential biomarker of ovarian cancer. We have solved x-ray structures of recombinant active hK7 at medium and atomic resolution in the presence of the inhibitors succinyl-Ala-Ala-Pro-Phe chloromethyl ketone and Ala-Ala-Phe-chloromethyl ketone. The most distinguishing features of hK7 are the short 70-80 loop and the unique S1 pocket, which prefers P1 Tyr residues, as shown by kinetic data. Similar to several other kallikreins, the enzyme activity is inhibited by Zn(2+) and Cu(2+) at low micromolar concentrations. Biochemical analyses of the mutants H99A and H41F confirm that only the metal-binding site at His(99) close to the catalytic triad accounts for the noncompetitive Zn(2+) inhibition type. Additionally, hK7 exhibits large positively charged surface patches, representing putative exosites for prime side substrate recognition. PMID- 17909183 TI - Stimulus-dependent differential regulation in the Escherichia coli PhoQ PhoP system. AB - In Escherichia coli, Salmonella, and related bacteria, the PhoQ-PhoP system regulates the expression of a large collection of genes in response to conditions of low magnesium or to the presence of certain antimicrobial peptides. We measured transcription of four PhoP-regulated promoters in E. coli that have significantly different PhoP-binding sites. Surprisingly, three promoters show identical responses to magnesium concentrations that range over four orders of magnitude. By analyzing and testing a simple model of transcriptional regulation, we find an explanation for this puzzle and show that these promoters are indeed differentially regulated at sufficiently high levels of stimulus. We then use this analysis to infer an effective level of phosphorylated PhoP as a function of magnesium stimulus. Our results demonstrate that differential regulation generally depends on the strength of the stimulus and highlight how quantitative analysis of stimulus-response curves can be used to infer properties of cell regulatory circuits that cannot be easily obtained from in vitro measurements. PMID- 17909182 TI - Kaposi's sarcoma herpesvirus-encoded latency-associated nuclear antigen stabilizes intracellular activated Notch by targeting the Sel10 protein. AB - Deregulation of the evolutionarily conserved Notch signaling is highly correlated with oncogenesis. Intracellular activated Notch (ICN) is a protooncogene linked to the transcription activation of a number of cellular genes involved in cell cycle regulation, differentiation, and proliferation. Stability of ICN is tightly regulated by the Sel10-mediated ubiquitin-proteasome pathway. Sel10 can function as a negative regulator of Notch and exhibits activities of a tumor-suppressor protein. This article shows that the Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) directly interacts with Sel10 and forms a complex in KSHV-infected cells. This results in suppression of ICN ubiquitination and degradation. The carboxyl terminus of LANA interacts with the F-box and WD40 domains of Sel10 and competes with ICN for binding to Sel10. This elevated level of ICN is also critical for maintaining the enhanced proliferation of KSHV-infected tumor cells. These findings describe a mechanism by which the KSHV-encoded LANA protein regulates ubiquitination of ICN mediated by the F-box component of the E3 ligase Sel10, leading to proliferation of the virus-infected cells. PMID- 17909184 TI - Heritability of ultimatum game responder behavior. AB - Experimental evidence suggests that many people are willing to deviate from materially maximizing strategies to punish unfair behavior. Even though little is known about the origins of such fairness preferences, it has been suggested that they have deep evolutionary roots and that they are crucial for maintaining and understanding cooperation among non-kin. Here we report the results of an ultimatum game, played for real monetary stakes, using twins recruited from the population-based Swedish Twin Registry as our subject pool. Employing standard structural equation modeling techniques, we estimate that >40% of the variation in subjects' rejection behavior is explained by additive genetic effects. Our estimates also suggest a very modest role for common environment as a source of phenotypic variation. Based on these findings, we argue that any attempt to explain observed ultimatum bargaining game behavior that ignores this genetic influence is incomplete. PMID- 17909185 TI - A chloroplast cyclophilin functions in the assembly and maintenance of photosystem II in Arabidopsis thaliana. AB - Photosynthetic light reactions rely on the proper function of large protein complexes (including photosystems I and II) that reside in the thylakoid membrane. Although their composition, structure, and function are known, the repertoire of assembly and maintenance factors is still being determined. Here we show that an immunophilin of the cyclophilin type, CYP38, plays a critical role in the assembly and maintenance of photosystem II (PSII) supercomplexes (SCs) in Arabidopsis. Mutant plants with the CYP38 gene interrupted by T-DNA insertion showed stunted growth and were hypersensitive to high light. Leaf chlorophyll fluorescence analysis and thylakoid membrane composition indicated that cyp38 mutant plants had defects in PSII SCs. Sucrose supplementation enabled the rescue of the mutant phenotype under low-light conditions, but failed to mitigate hypersensitivity to high-light stress. Protein radiolabeling assays showed that, although individual thylakoid proteins were synthesized equally in mutant and wild type, the assembly of the PSII SC was impaired in the mutant. In addition, the D1 and D2 components of the mutant PSII had a short half-life under high light stress. The results provide evidence that CYP38 is necessary for the assembly and stabilization of PSII. PMID- 17909186 TI - Traffic related exposures and lung function in adults. PMID- 17909187 TI - Where there's smoke there's lung disease. PMID- 17909188 TI - Is it time to change the approach to oxygen therapy in the breathless patient? PMID- 17909189 TI - Challenges in pulmonary fibrosis . 4: smoking-induced diffuse interstitial lung diseases. AB - Smoking-induced diffuse interstitial lung processes include respiratory bronchiolitis, respiratory bronchiolitis-associated interstitial lung disease (RBILD), desquamative interstitial pneumonia (DIP) and Langerhans' cell histiocytosis. The histological, radiological and clinical features of respiratory bronchiolitis, RBILD and DIP are reviewed, with particular reference to management issues; Langerhans' cell histiocytosis is covered elsewhere in this series of articles. Possible relationships between smoking and other diffuse lung diseases are explored briefly. PMID- 17909191 TI - Which children with persistent cough should be regarded as persistent bacterial bronchitis? PMID- 17909190 TI - Central hypoventilation with PHOX2B expansion mutation presenting in adulthood. AB - Congenital central hypoventilation syndrome most commonly presents in neonates with sleep related hypoventilation; late onset cases have occurred up to the age of 10 years. It is associated with mutations in the PHOX2B gene, encoding a transcription factor involved in autonomic nervous system development. The case history is described of an adult who presented with chronic respiratory failure due to PHOX2B mutation-associated central hypoventilation and an impaired response to hypercapnia. PMID- 17909193 TI - HIV-related TB and adverse drug events. PMID- 17909192 TI - Outcomes in children treated for persistent bacterial bronchitis. PMID- 17909194 TI - Diagnosis of COPD. PMID- 17909195 TI - Is childhood immunisation associated with atopic disease from age 7 to 32 years? PMID- 17909196 TI - Reducing door-to-antibiotic time in community acquired pneumonia. PMID- 17909197 TI - Donor lymphocyte infusion in the treatment of first hematological relapse after allogeneic stem-cell transplantation in adults with acute myeloid leukemia: a retrospective risk factors analysis and comparison with other strategies by the EBMT Acute Leukemia Working Party. AB - PURPOSE: To evaluate the role of donor lymphocyte infusion (DLI) in the treatment of relapsed acute myeloid leukemia (AML) after allogeneic hematopoietic stem cell transplantation (HSCT). PATIENTS AND METHODS: We retrospectively analyzed the data of 399 patients with AML in first hematological relapse after HSCT whose treatment did (n = 171) or did not (n = 228) include DLI. After correction for imbalances and established risk factors, the two groups were compared with respect to overall survival. Further, a detailed analysis of risk factors for survival among DLI recipients was performed. RESULTS: Median follow-up was 27 and 40 months, respectively. Estimated survival at 2 years (+/- standard deviation) was 21% +/- 3% for patients receiving DLI and 9% +/- 2% for patients not receiving DLI. After adjustment for differences between the groups, better outcome was associated with age younger than 37 years (P = .008), relapse occurring more than 5 months after HSCT (P < .0001), and use of DLI (P = .04). Among DLI recipients, a lower tumor burden at relapse (< 35% of bone marrow blasts; P = .006), female sex (P = .02), favorable cytogenetics (P = .004), and remission at time of DLI (P < .0001) were predictive for survival in a multivariate analysis. Two-year survival was 56% +/- 10%, if DLI was performed in remission or with favorable karyotype, and 15% +/- 3% if DLI was given in aplasia or with active disease. CONCLUSION: Although further evidence for a graft-versus leukemia effect by DLI is provided, our results confirm, that the clinical benefit is limited to a minority of patients. Strategies to reduce tumor burden before DLI, as well as alternative treatment options should be investigated in adults with relapsed AML after HSCT. PMID- 17909198 TI - Antifungal prophylaxis in cancer patients after chemotherapy or hematopoietic stem-cell transplantation: systematic review and meta-analysis. AB - PURPOSE: To evaluate the effect of antifungal prophylaxis on all-cause mortality as primary outcome, invasive fungal infections (IFIs), and adverse events. Many studies have evaluated the role of antifungal prophylaxis in cancer patients, with inconsistent conclusions. METHODS: We performed a systematic review and meta analysis of randomized, controlled trials comparing systemic antifungals with placebo, no intervention, or other antifungal agents for prophylaxis in cancer patients after chemotherapy. The Cochrane Library, MEDLINE, conference proceedings, and references were searched. Two reviewers independently appraised the quality of trials and extracted data. RESULTS: Sixty-four trials met inclusion criteria. Antifungal prophylaxis decreased all-cause mortality significantly at end of follow-up compared with placebo, no treatment, or nonsystemic antifungals (relative risk [RR], 0.84; 95% CI, 0.74 to 0.95). In allogeneic hematopoietic stem-cell transplantation (HSCT) recipients, prophylaxis reduced all-cause mortality (RR, 0.62; 95% CI, 0.45 to 0.85), fungal-related mortality, and documented IFI. In acute leukemia patients, there was a significant reduction in fungal-related mortality and documented IFI, whereas the difference in mortality was only borderline significant (RR, 0.88; 95% CI, 0.74 to 1.06). Prophylaxis with itraconazole suspension reduced documented IFI when compared with fluconazole, with no difference in survival, and at the cost of more adverse events. On the basis of two studies, posaconazole prophylaxis reduced all-cause mortality (RR, 0.74; 95% CI, 0.56 to 0.98), fungal-related mortality, and IFI when compared with fluconazole. CONCLUSION: Antifungal prophylaxis decreases all-cause mortality significantly in patients after chemotherapy. Antifungal prophylaxis should be administered to patients undergoing allogeneic HSCT, and should probably be administered to high-risk acute leukemia patients. PMID- 17909199 TI - Phase II study of efficacy and safety of bevacizumab in combination with chemotherapy or erlotinib compared with chemotherapy alone for treatment of recurrent or refractory non small-cell lung cancer. AB - PURPOSE: Bevacizumab, a humanized anti-vascular endothelial growth factor monoclonal antibody, and erlotinib, a reversible, orally available epidermal growth factor receptor tyrosine kinase inhibitor, have demonstrated evidence of a survival benefit in the treatment of non-small-cell lung cancer (NSCLC). A single arm phase I and II study of bevacizumab plus erlotinib demonstrated encouraging efficacy, with a favorable safety profile. PATIENTS AND METHODS: A multicenter, randomized phase II trial evaluated the safety of combining bevacizumab with either chemotherapy (docetaxel or pemetrexed) or erlotinib and preliminarily assessed these combinations versus chemotherapy alone, as measured by progression free survival (PFS). All patients had histologically confirmed nonsquamous NSCLC that had progressed during or after one platinum-based regimen. RESULTS: One hundred twenty patients were randomly assigned and treated. No unexpected adverse events were noted. Fewer patients (13%) in the bevacizumab-erlotinib arm discontinued treatment as a result of adverse events than in the chemotherapy alone (24%) or bevacizumab-chemotherapy (28%) arms. The incidence of grade 5 hemorrhage in patients receiving bevacizumab was 5.1%. Although not statistically significant, relative to chemotherapy alone, the risk of disease progression or death was 0.66 (95% CI, 0.38 to 1.16) among patients treated with bevacizumab chemotherapy and 0.72 (95% CI, 0.42 to 1.23) among patients treated with bevacizumab-erlotinib. One-year survival rate was 57.4% for bevacizumab-erlotinib and 53.8% for bevacizumab-chemotherapy compared with 33.1% for chemotherapy alone. CONCLUSION: Results for PFS and overall survival favor combination of bevacizumab with either chemotherapy or erlotinib over chemotherapy alone in the second-line setting. No unexpected safety signals were noted. The rate of fatal pulmonary hemorrhage was consistent with previous bevacizumab trials. The toxicity profile of the bevacizumab-erlotinib combination is favorable compared with either chemotherapy-containing group. PMID- 17909200 TI - Summaries for patients. Antiviral therapy for chronic hepatitis B. PMID- 17909202 TI - Summaries for patients. Validity of models for predicting BRCA1 and BRCA2 mutations. PMID- 17909201 TI - Treatment of hepatitis B e antigen positive chronic hepatitis with telbivudine or adefovir: a randomized trial. AB - BACKGROUND: The efficacy of nucleoside and nucleotide analogues for hepatitis B has been linked to the magnitude and durability of hepatitis B virus (HBV) suppression. OBJECTIVE: To compare the antiviral efficacy of telbivudine and adefovir dipivoxil, and the effects of switching from adefovir to telbivudine, in hepatitis B e antigen (HBeAg)-positive patients with chronic hepatitis B. DESIGN: Randomized, controlled, open-label trial. SETTING: 16 outpatient clinics. PATIENTS: 135 treatment-naive, HBeAg-positive adults with chronic hepatitis B. INTERVENTION: Patients were randomly assigned in a 1:1:1 ratio to 52 weeks of telbivudine (group A) or adefovir (group B), or 24 weeks of adefovir and then telbivudine for the remaining 28 weeks (group C). One hundred thirty-one patients completed 52 weeks of treatment. MEASUREMENTS: The primary efficacy comparison was serum HBV DNA reduction at week 24, with a secondary comparison at week 52. RESULTS: At week 24, mean HBV DNA reduction was greater in group A than in pooled groups B and C (-6.30 vs. -4.97 log10 copies/mL; difference, -1.33 log10 copies/mL [95% CI, -1.99 to -0.66 log(10) copies/mL]; P < 0.001), and more patients in group A were polymerase chain reaction-negative (39% vs. 12%; odds ratio, 4.46 [CI, 1.86 to 10.72]; P = 0.001). At week 52, the mean residual HBV DNA level was lower in group A and group C than in group B (3.01 log10 copies/mL [group A] and 3.02 log10 copies/mL [group C] vs. 4.00 log10 copies/mL [group B]; difference, -0.99 log10 copies/mL [CI, -1.67 to -0.32 log10 copies/mL] and -0.98 log10 copies/mL [CI, -1.64 to -0.32 log10 copies/mL]; P = 0.004). Adverse events were similar across groups; the most common were upper respiratory symptoms, headache, back pain, and diarrhea. LIMITATIONS: The trial was open-label and was not of sufficient size or duration to compare clinical outcomes and long-term efficacy. CONCLUSION: Telbivudine demonstrated greater and more consistent HBV DNA suppression than adefovir after 24 weeks of treatment. After 52 weeks, HBV DNA suppression was greater in patients who had received continuous telbivudine or were switched to telbivudine after 24 weeks than in those who received continuous adefovir. PMID- 17909203 TI - Summaries for patients. Diagnosis and treatment of low back pain: recommendations from the American College of Physicians/American Pain Society. PMID- 17909204 TI - In the clinic. Influenza. PMID- 17909207 TI - Cost-effectiveness of screening and vaccinating Asian and Pacific Islander adults for hepatitis B. AB - BACKGROUND: As many as 10% of Asian and Pacific Islander adults in the United States are chronically infected with hepatitis B virus (HBV), and up to two thirds are unaware that they are infected. Without proper medical management and antiviral therapy, up to 25% of Asian and Pacific Islander persons with chronic HBV infection will die of liver disease. OBJECTIVE: To assess the cost effectiveness of 4 HBV screening and vaccination programs for Asian and Pacific Islander adults in the United States. DESIGN: Markov model with costs and benefits discounted at 3%. DATA SOURCES: Published literature and expert opinion. TARGET POPULATION: Asian and Pacific Islander adults (base-case age, 40 years; sensitivity analysis conducted on ages 20 to 60 years). TIME HORIZON: Lifetime. PERSPECTIVE: U.S. societal. INTERVENTIONS: A universal vaccination strategy in which all individuals are given a 3-dose vaccination series; a screen-and-treat strategy, in which individuals are given blood tests to determine whether they are chronically infected, and infected persons are monitored and treated; a screen, treat, and ring vaccinate strategy, in which all individuals are tested for chronic HBV infection and close contacts of infected persons are screened and vaccinated if needed; and a screen, treat, and vaccinate strategy, in which all individuals are tested and then vaccinated with a 3-dose series if needed. In all cases, persons found to be chronically infected are monitored and treated if indicated. OUTCOME MEASURES: Costs (2006 U.S. dollars), quality-adjusted life years (QALYs), and incremental cost-effectiveness. RESULTS OF BASE-CASE ANALYSIS: Compared with the status quo, the screen-and-treat strategy has an incremental cost-effectiveness ratio of $36,088 per QALY gained. The screen, treat, and ring vaccinate strategy gains more QALYs than the screen and treat strategy and incurs modest incremental costs, leading to incremental cost-effectiveness of $39,903 per QALY gained compared with the screen and treat strategy. The universal vaccination and screen, treat, and vaccinate strategies were weakly dominated by the other 2 strategies. RESULTS OF SENSITIVITY ANALYSIS: Over a wide range of variables, the incremental cost-effectiveness ratios of the screen and treat and screen, treat, and ring vaccinate strategies were less than $50,000 per QALY gained. LIMITATIONS: Results depend on the accuracy of the underlying data and assumptions. The long-term effectiveness of new and future HBV treatments is uncertain. CONCLUSIONS: Screening programs for HBV among Asian and Pacific Islander adults are likely to be cost effective. Clinically significant benefits accrue from identifying chronically infected persons for medical management and vaccinating their close contacts. Such efforts can greatly reduce the burden of HBV-associated liver cancer and chronic liver disease in the Asian and Pacific Islander population. PMID- 17909206 TI - Serum and biliary insulin-like growth factor I and vascular endothelial growth factor in determining the cause of obstructive cholestasis. AB - BACKGROUND: Cholangiocarcinoma cells express and secrete insulin-like growth factor I (IGF-I) and vascular endothelial growth factor (VEGF). OBJECTIVE: To measure IGF-I and VEGF in bile and serum of patients with extrahepatic cholangiocarcinoma and to evaluate their performance as diagnostic markers. DESIGN: Cross-sectional study. SETTING: Inpatients at the Division of Gastroenterology, University Hospital, Ancona, Italy. PATIENTS: 73 patients who consecutively had endoscopic retrograde cholangiopancreatography (ERCP), including patients with extrahepatic cholangiocarcinoma (n = 29), pancreatic cancer (n = 19), and benign biliary abnormalities (n = 25; bile duct stones, primary sclerosing cholangitis, and cholangitis). MEASUREMENTS: Diagnosis was based on conventional radiology, ERCP, and follow-up. Insulin-like growth factor I and VEGF were measured by using enzyme-linked immunosorbent assay. RESULTS: The biliary IGF-I concentration was 15- to 20-fold higher (P < 0.001) in extrahepatic cholangiocarcinoma (mean, 84.6 nmol/L [95% CI, 74.0 to 95.2 nmol/L]) than in pancreatic cancer (5.8 nmol/L [CI, 4.0 to 7.5 nmol/L]) or benign biliary abnormalities (4.1 nmol/L [CI, 3.1 to 5.2 nmol/L]). The area under the receiver operating characteristic curve was 1 when biliary IGF-I values in the extrahepatic cholangiocarcinoma were compared with benign biliary abnormalities or pancreatic cancer. In contrast, biliary VEGF concentration was similar in the 3 groups. Serum IGF-I levels were similar among the groups, whereas serum VEGF levels were higher in the cholangiocarcinoma (0.97 ng/mL [CI, 0.59 to 1.35 ng/mL]; P = 0.0016) and pancreatic cancer groups (0.66 ng/mL [CI, 0.43 to 0.88 ng/mL]; P < 0.001) compared with patients with benign biliary abnormalities (0.28 ng/mL [CI, 0.17 to 0.37 ng/mL]). LIMITATIONS: Data were obtained in a small sample, the study was performed in a single center, and few patients had a tissue diagnosis. CONCLUSIONS: Biliary IGF-I levels in patients undergoing ERCP for biliary obstruction may differentiate extrahepatic cholangiocarcinoma from either pancreatic cancer or benign biliary abnormalities. PMID- 17909205 TI - Validity of models for predicting BRCA1 and BRCA2 mutations. AB - BACKGROUND: Deleterious mutations of the BRCA1 and BRCA2 genes confer susceptibility to breast and ovarian cancer. At least 7 models for estimating the probabilities of having a mutation are used widely in clinical and scientific activities; however, the merits and limitations of these models are not fully understood. OBJECTIVE: To systematically quantify the accuracy of the following publicly available models to predict mutation carrier status: BRCAPRO, family history assessment tool, Finnish, Myriad, National Cancer Institute, University of Pennsylvania, and Yale University. DESIGN: Cross-sectional validation study, using model predictions and BRCA1 or BRCA2 mutation status of patients different from those used to develop the models. SETTING: Multicenter study across Cancer Genetics Network participating centers. PATIENTS: 3 population-based samples of participants in research studies and 8 samples from genetic counseling clinics. MEASUREMENTS: Discrimination between individuals testing positive for a mutation in BRCA1 or BRCA2 from those testing negative, as measured by the c-statistic, and sensitivity and specificity of model predictions. RESULTS: The 7 models differ in their predictions. The better-performing models have a c-statistic around 80%. BRCAPRO has the largest c-statistic overall and in all but 2 patient subgroups, although the margin over other models is narrow in many strata. Outside of high-risk populations, all models have high false-negative and false positive rates across a range of probability thresholds used to refer for mutation testing. LIMITATION: Three recently published models were not included. CONCLUSIONS: All models identify women who probably carry a deleterious mutation of BRCA1 or BRCA2 with adequate discrimination to support individualized genetic counseling, although discrimination varies across models and populations. PMID- 17909208 TI - Update in geriatric medicine. PMID- 17909209 TI - Diagnosis and treatment of low back pain: a joint clinical practice guideline from the American College of Physicians and the American Pain Society. AB - RECOMMENDATION 1: Clinicians should conduct a focused history and physical examination to help place patients with low back pain into 1 of 3 broad categories: nonspecific low back pain, back pain potentially associated with radiculopathy or spinal stenosis, or back pain potentially associated with another specific spinal cause. The history should include assessment of psychosocial risk factors, which predict risk for chronic disabling back pain (strong recommendation, moderate-quality evidence). RECOMMENDATION 2: Clinicians should not routinely obtain imaging or other diagnostic tests in patients with nonspecific low back pain (strong recommendation, moderate-quality evidence). RECOMMENDATION 3: Clinicians should perform diagnostic imaging and testing for patients with low back pain when severe or progressive neurologic deficits are present or when serious underlying conditions are suspected on the basis of history and physical examination (strong recommendation, moderate-quality evidence). RECOMMENDATION 4: Clinicians should evaluate patients with persistent low back pain and signs or symptoms of radiculopathy or spinal stenosis with magnetic resonance imaging (preferred) or computed tomography only if they are potential candidates for surgery or epidural steroid injection (for suspected radiculopathy) (strong recommendation, moderate-quality evidence). RECOMMENDATION 5: Clinicians should provide patients with evidence-based information on low back pain with regard to their expected course, advise patients to remain active, and provide information about effective self-care options (strong recommendation, moderate-quality evidence). RECOMMENDATION 6: For patients with low back pain, clinicians should consider the use of medications with proven benefits in conjunction with back care information and self-care. Clinicians should assess severity of baseline pain and functional deficits, potential benefits, risks, and relative lack of long-term efficacy and safety data before initiating therapy (strong recommendation, moderate-quality evidence). For most patients, first-line medication options are acetaminophen or nonsteroidal anti-inflammatory drugs. RECOMMENDATION 7: For patients who do not improve with self-care options, clinicians should consider the addition of nonpharmacologic therapy with proven benefits-for acute low back pain, spinal manipulation; for chronic or subacute low back pain, intensive interdisciplinary rehabilitation, exercise therapy, acupuncture, massage therapy, spinal manipulation, yoga, cognitive-behavioral therapy, or progressive relaxation (weak recommendation, moderate-quality evidence). PMID- 17909210 TI - Nonpharmacologic therapies for acute and chronic low back pain: a review of the evidence for an American Pain Society/American College of Physicians clinical practice guideline. AB - BACKGROUND: Many nonpharmacologic therapies are available for treatment of low back pain. PURPOSE: To assess benefits and harms of acupuncture, back schools, psychological therapies, exercise therapy, functional restoration, interdisciplinary therapy, massage, physical therapies (interferential therapy, low-level laser therapy, lumbar supports, shortwave diathermy, superficial heat, traction, transcutaneous electrical nerve stimulation, and ultrasonography), spinal manipulation, and yoga for acute or chronic low back pain (with or without leg pain). DATA SOURCES: English-language studies were identified through searches of MEDLINE (through November 2006) and the Cochrane Database of Systematic Reviews (2006, Issue 4). These electronic searches were supplemented by hand searching of reference lists and additional citations suggested by experts. STUDY SELECTION: Systematic reviews and randomized trials of 1 or more of the preceding therapies for acute or chronic low back pain (with or without leg pain) that reported pain outcomes, back-specific function, general health status, work disability, or patient satisfaction. DATA EXTRACTION: We abstracted information about study design, population characteristics, interventions, outcomes, and adverse events. To grade methodological quality, we used the Oxman criteria for systematic reviews and the Cochrane Back Review Group criteria for individual trials. DATA SYNTHESIS: We found good evidence that cognitive behavioral therapy, exercise, spinal manipulation, and interdisciplinary rehabilitation are all moderately effective for chronic or subacute (>4 weeks' duration) low back pain. Benefits over placebo, sham therapy, or no treatment averaged 10 to 20 points on a 100-point visual analogue pain scale, 2 to 4 points on the Roland-Morris Disability Questionnaire, or a standardized mean difference of 0.5 to 0.8. We found fair evidence that acupuncture, massage, yoga (Viniyoga), and functional restoration are also effective for chronic low back pain. For acute low back pain (<4 weeks' duration), the only nonpharmacologic therapies with evidence of efficacy are superficial heat (good evidence for moderate benefits) and spinal manipulation (fair evidence for small to moderate benefits). Although serious harms seemed to be rare, data on harms were poorly reported. No trials addressed optimal sequencing of therapies, and methods for tailoring therapy to individual patients are still in early stages of development. Evidence is insufficient to evaluate the efficacy of therapies for sciatica. LIMITATIONS: Our primary source of data was systematic reviews. We included non-English language trials only if they were included in English-language systematic reviews. CONCLUSIONS: Therapies with good evidence of moderate efficacy for chronic or subacute low back pain are cognitive-behavioral therapy, exercise, spinal manipulation, and interdisciplinary rehabilitation. For acute low back pain, the only therapy with good evidence of efficacy is superficial heat. PMID- 17909212 TI - Cancer risk models: translating family history into clinical management. PMID- 17909211 TI - Medications for acute and chronic low back pain: a review of the evidence for an American Pain Society/American College of Physicians clinical practice guideline. AB - BACKGROUND: Medications are the most frequently prescribed therapy for low back pain. A challenge in choosing pharmacologic therapy is that each class of medication is associated with a unique balance of risks and benefits. PURPOSE: To assess benefits and harms of acetaminophen, nonsteroidal anti-inflammatory drugs (NSAIDs), antidepressants, benzodiazepines, antiepileptic drugs, skeletal muscle relaxants, opioid analgesics, tramadol, and systemic corticosteroids for acute or chronic low back pain (with or without leg pain). DATA SOURCES: English-language studies were identified through searches of MEDLINE (through November 2006) and the Cochrane Database of Systematic Reviews (2006, Issue 4). These electronic searches were supplemented by hand searching reference lists and additional citations suggested by experts. STUDY SELECTION: Systematic reviews and randomized trials of dual therapy or monotherapy with 1 or more of the preceding medications for acute or chronic low back pain that reported pain outcomes, back specific function, general health status, work disability, or patient satisfaction. DATA EXTRACTION: We abstracted information about study design, population characteristics, interventions, outcomes, and adverse events. To grade methodological quality, we used the Oxman criteria for systematic reviews and the Cochrane Back Review Group criteria for individual trials. DATA SYNTHESIS: We found good evidence that NSAIDs, acetaminophen, skeletal muscle relaxants (for acute low back pain), and tricyclic antidepressants (for chronic low back pain) are effective for pain relief. The magnitude of benefit was moderate (effect size of 0.5 to 0.8, improvement of 10 to 20 points on a 100-point visual analogue pain scale, or relative risk of 1.25 to 2.00 for the proportion of patients experiencing clinically significant pain relief), except in the case of tricyclic antidepressants (for which the benefit was small to moderate). We also found fair evidence that opioids, tramadol, benzodiazepines, and gabapentin (for radiculopathy) are effective for pain relief. We found good evidence that systemic corticosteroids are ineffective. Adverse events, such as sedation, varied by medication, although reliable data on serious and long-term harms are sparse. Most trials were short term (< or =4 weeks). Few data address efficacy of dual-medication therapy compared with monotherapy, or beneficial effects on functional outcomes. LIMITATIONS: Our primary source of data was systematic reviews. We included non-English-language trials only if they were included in English-language systematic reviews. CONCLUSIONS: Medications with good evidence of short-term effectiveness for low back pain are NSAIDs, acetaminophen, skeletal muscle relaxants (for acute low back pain), and tricyclic antidepressants (for chronic low back pain). Evidence is insufficient to identify one medication as offering a clear overall net advantage because of complex tradeoffs between benefits and harms. Individual patients are likely to differ in how they weigh potential benefits, harms, and costs of various medications. PMID- 17909213 TI - The story between the pinstripes: interviewing for internal medicine residencies. PMID- 17909214 TI - Management of venous thromboembolism. PMID- 17909215 TI - Management of venous thromboembolism. PMID- 17909216 TI - Management of venous thromboembolism. PMID- 17909217 TI - Intensive intraoperative insulin therapy versus conventional glucose management during cardiac surgery. PMID- 17909218 TI - Intensive intraoperative insulin therapy versus conventional glucose management during cardiac surgery. PMID- 17909219 TI - Intensive intraoperative insulin therapy versus conventional glucose management during cardiac surgery. PMID- 17909220 TI - Quantitative immunochemical fecal occult blood test for diagnosing colorectal neoplasia. PMID- 17909221 TI - Anticoagulant prophylaxis for hospitalized medical patients. PMID- 17909222 TI - Psychosocial influences on suboptimal adjuvant breast cancer treatment adherence among African American women: implications for education and intervention. AB - Despite lower incidence, African American women are at increased risk of dying from breast cancer relative to their European American counterparts. Although there are key differences in both screening behavior and tumor characteristics, an additional part of this mortality difference may lie in the fact that African American women receive suboptimal adjuvant chemotherapy and may receive suboptimal hormonal therapy, therapies that are known to increase survival. The authors consider ethnic differences in the psychosocial factors that have been shown to relate to poor screening adherence and consider how they may influence adherence to breast cancer adjuvant treatment, thus the receipt of suboptimal adjuvant chemo or hormonal therapy. To this end, they review ethnic differences in cognitive, emotional, and social network variables. Psychosocial variables should be included in research designed to understand cancer disparities as well interventions that can be tailored to culturally diverse populations to improve treatment adherence. PMID- 17909224 TI - Methionine restriction effects on 11 -HSD1 activity and lipogenic/lipolytic balance in F344 rat adipose tissue. AB - Methionine restriction (MR) limits age-related adiposity in Fischer 344 (F344) rats. To assess the mechanism of adiposity resistance, the effect of MR on adipose tissue (AT) 11beta-hydroxysteroid dehydrogenase-1 (11beta-HSD1) was examined. MR induced 11beta-HSD1 activity in all ATs, correlating with increased tissue corticosterone. However, an inverse relationship between 11beta-HSD1 activity and adipocyte size was observed. Because dietary restriction controls lipogenic and lipolytic rates, MR's effects on lipogenic and lipolytic enzymes were evaluated. MR increased adipose triglyceride lipase and acetyl-coenzyme A carboxylase (ACC) protein levels but induced ACC phosphorylation at serine residues that render the enzyme inactive, suggesting alterations of basal lipolysis and lipogenesis. In contrast, no changes in basal or phosphorylated hormone-sensitive lipase levels were observed. ACC-phosphorylated sites were specific for AMP-activated protein kinase (AMPK); therefore, AMPK activation was evaluated. Significant differences in AMPKalpha protein, phosphorylation, and activity levels were observed only in retroperitoneal fat from MR rats. No differences in protein kinase A phosphorylation and intracellular cAMP levels were detected. In vitro studies revealed increased lipid degradation and a trend toward increased lipid synthesis, suggesting the presence of a futile cycle. In conclusion, MR disrupts the lipogenic/lipolytic balance, contributing importantly to adiposity resistance in F344 rats. PMID- 17909223 TI - Electronegative LDL circulating in smokers impairs endothelial progenitor cell differentiation by inhibiting Akt phosphorylation via LOX-1. AB - Endothelial progenitor cells (EPCs), important for endothelial regeneration and vasculogenesis, are reduced by cigarette smoking. To elucidate the mechanisms, we examined the effects of electronegative LDL, circulating in chronic smokers, on EPC differentiation. Using ion-exchange chromatography, we purified smoker LDL into five subfractions, L1-L5. In matched, nonsmoking healthy subjects, L5, the most electronegative subfraction, was either absent or scanty. Sustained L5 treatment inhibited CD31 and KDR expression and EPC differentiation, whereas L1 L4 had no effect. L5 also inhibited telomerase activity to accelerate EPC senescence in correlation with reduced Akt phosphorylation. Transfection of day 3 EPCs with dominant negative Akt constructs inhibited CD31 and KDR expression, stalled EPC differentiation, and promoted early senescence. In contrast, transfection with constitutively active Akt rendered the EPCs resistant to L5, allowing normal maturation. L5 upregulated the lectin-like oxidized low density lipoprotein receptor 1 (LOX-1), and pretreatment of EPCs with TS20, a LOX-1 neutralizing antibody, blocked internalization of L5 by EPCs and prevented L5 mediated inhibition of EPC differentiation. Mixing L5 with L1 to physiological L5/L1 ratios did not attenuate L5's effects. These findings suggest that cigarette smoking is associated with the formation of L5, which inhibits EPC differentiation by impairing Akt phosphorylation via the LOX-1 receptor. PMID- 17909225 TI - Putting evidence into context: some advice for guideline writers. PMID- 17909226 TI - Practice corner: chair's rounds. PMID- 17909227 TI - Long term use of eicosapentaenoic acid reduced major coronary events in hypercholesterolaemia. PMID- 17909228 TI - Hormone therapy for younger women may not increase CHD risk during 5 7 years follow-up, but stroke risk was increased independent of age. PMID- 17909229 TI - The Atkins diet led to more weight loss than the Zone diet in overweight and obese premenopausal women at 12 months. PMID- 17909230 TI - A care management intervention improved depression after stroke. PMID- 17909231 TI - A self-management approach to patient education for type 2 diabetes was more effective than a didactic approach. PMID- 17909232 TI - Carvedilol was more effective than metoprolol for preventing cardiovascular events in heart failure. PMID- 17909233 TI - An escalating higher energy regimen was better than a fixed lower energy regimen for defibrillation in out of hospital cardiac arrest. PMID- 17909234 TI - 6 or 3 months of anticoagulant therapy did not differ for treatment failure in patients with DVT, PE, or both. PMID- 17909235 TI - Ibuprofen was more effective than codeine or acetaminophen for musculoskeletal pain in children. PMID- 17909236 TI - A once yearly IV infusion of zoledronic acid prevented fractures in postmenopausal women with osteoporosis. PMID- 17909237 TI - 10 day sequential therapy was more effective than 10 day triple drug therapy for eradicating Helicobacter pylori infection. PMID- 17909238 TI - Review: partner notification interventions can reduce sexually transmitted infections. PMID- 17909239 TI - Ambulatory titration of continuous positive airway pressure was as effective as polysomnography for obstructive sleep apnoea. PMID- 17909241 TI - A short version of Mini-Mental State Examination (MMSE) was as accurate as the original MMSE for predicting dementia. PMID- 17909240 TI - The 2-item Generalized Anxiety Disorder scale had high sensitivity and specificity for detecting GAD in primary care. PMID- 17909242 TI - Review: clinical examination is often as accurate as magnetic resonance imaging for diagnosing meniscus tears. PMID- 17909243 TI - Review: glycated haemoglobin A1c and fasting plasma glucose screening tests have similar sensitivities and specificities for early detection of type 2 diabetes. PMID- 17909244 TI - Routine rotavirus vaccinations of infants cost US$138 per case averted and US$197,190 per life year saved from a societal perspective. PMID- 17909245 TI - A clinical prediction guide predicted progression to rheumatoid arthritis in undifferentiated arthritis. PMID- 17909246 TI - Pre-endoscopic serological test with duodenal biopsy in high risk patients had high sensitivity and low specificity for coeliac disease. PMID- 17909247 TI - Learning disabilities and delinquency: a study of Israeli prison inmates. AB - Research consistently illustrates that several intellectual disabilities--namely, learning disabilities (LD), low intelligence, challenging behavior, and inadequate adaptive behavior, as well as attention deficit hyperactivity disorder (ADHD)--are considered risk factors for antisocial and criminal behavior. Although much attention has been paid to the relationship of LD, ADHD, and criminal behavior, three research topics have been overlooked: the frequency of LD with ADHD among inmates, the relationship between LD and/or ADHD and level of education among prisoners, and the connection between LD and/or ADHD and age of criminal onset. The present study examined the frequency of LD and ADHD in a sample of Israeli-born prisoners, in addition to the frequency of each category by itself, and it investigated the relationship of LD and/or ADHD, school dropout age, and onset of criminal activity. PMID- 17909255 TI - 18F-DOPA PET and PET/CT. PMID- 17909256 TI - 18F-FDG PET in non-Hodgkin's lymphoma: qualitative or quantitative? PMID- 17909257 TI - PET imaging of CCND1 mRNA in human MCF7 estrogen receptor positive breast cancer xenografts with oncogene-specific [64Cu]chelator-peptide nucleic acid-IGF1 analog radiohybridization probes. AB - Treatment of breast cancer is hampered by a large unmet need for rapid, sensitive, specific staging and stratification of palpable and nonpalpable abnormalities. Mammography and physical examination miss many early breast cancers, yet detect many benign lesions. Cyclin D1, encoded by CCND1 messenger RNA (mRNA), and insulin-like growth factor 1 receptor (IGF1R) are key regulators of cell proliferation that are overexpressed in most breast cancers. Therefore, we hypothesized that malignant breast masses could be imaged and quantitated externally by PET with a dual-specificity probe that targets both CCND1 mRNA and IGF1R. METHODS: We designed a CCND1-specific peptide nucleic acid (PNA) hybridization sequence (CTGGTGTTCCAT), separated by a C-terminal spacer to a cyclized IGF1 peptide analog (d-Cys-Ser-Lys-Cys), for IGF1R-mediated endocytosis. On the N-terminus we attached a chelator (1,4,7 tris(carboxymethylaza)cyclododecane-10-azaacetyl [DO3A]) for the positron emitting nuclide (64)Cu. We administered the [(64)Cu]CCND1-IGF1 analog radiohybridization probes, as well as sequence controls, by tail vein to immunocompromised female NCr mice bearing human MCF7 estrogen-dependent, receptor positive xenografts. We imaged the mice by PET and CT 4 and 24 h later, and measured tissue distribution of the radiohybridization probes. RESULTS: We observed 8 +/- 2-fold higher PET intensity in the center of the breast cancer xenografts than in the contralateral tissues at 24 h after injection of the [(64)Cu]CCND1-IGF1 analog radiohybridization probe. IGF1 blocking yielded significantly weaker images (P < 0.05) relative to the tumor-free side at 24 h after injection, as did a PNA mismatch probe, a peptide mismatch probe, and free (64)CuCl(2). CONCLUSION: These results are consistent with our hypothesis for radiohybridization PET of overexpressed CCND1 mRNA, dependent on IGF1R-mediated endocytosis, in suspect masses. Early noninvasive detection of initial cancerous transformation, as well as invasive or recurrent breast cancer, with dual specificity radiohybridization probes, might enable molecularly targeted staging, stratification, and choice of therapy. PMID- 17909258 TI - Dynamic tracking during intracoronary injection of 18F-FDG-labeled progenitor cell therapy for acute myocardial infarction. AB - We assessed the feasibility of dynamic 3-dimensional (3D) PET/CT tracking of (18)F-FDG-labeled circulating progenitor cell (CPC) therapy during intracoronary injection, using a porcine model of acute myocardial infarction (MI). METHODS: Human and porcine CPC were radiolabeled with (18)F-FDG, with variation in temperature and incubation time to determine optimal conditions. For in vivo experiments, CPC were harvested before induction of infarction (using 90-min coronary balloon occlusion). At 48 h, animals underwent cardiac MRI to assess infarct size. A balloon catheter was placed in the infarct artery at the same location as that used for induction of MI, and during dynamic 3D PET/CT 3 x 10(7) autologous (18)F-FDG progenitor cells were injected through the central lumen using either (a) 3 cycles of balloon occlusion and reperfusion or (b) high concentration, single-bolus injection without balloon occlusion (n = 3 for both protocols). Peripheral blood was drawn at 1-min intervals during cell injection. RESULTS: Labeling efficiency was optimized by 30-min incubation at 37 degrees C (human CPC, 89.9% +/- 4.8%; porcine CPC, 91.6% +/- 6.4%). Cell-bound activity showed a nonsignificant decrease at 1 h (human, 74.3% +/- 10.7%; porcine, 77.7% +/- 12.8%; P > 0.05) and a significant decrease at 2 h (human, 62.1% +/- 8.9%; porcine, 68.6% +/- 5.4%; P = 0.009). Mean infarct size was similar for both injection protocols (16.3% +/- 3.4% and 20.6% +/- 2.7%; P > 0.05). Dynamic scanning demonstrated a sharp rise in myocardial activity during each cycle of balloon-occlusion cell delivery, with a significant fall in activity (around 80%) immediately after balloon deflation. The latter was associated with a transient spike in peripheral blood (18)F-FDG activity, consistent with the first pass of labeled cells in the systemic circulation. A single spike and gradual fall in myocardial activity was observed with high-concentration, single-bolus therapy. At 1 h, myocardial activity was 8.7% +/- 1.5% of total injected dose for balloon occlusion delivery and 17.8% +/- 7.9% for high-concentration, single-bolus delivery (P = 0.08). CONCLUSION: Dynamic tracking during intracoronary injection of (18)F-FDG-labeled CPC is feasible and demonstrates significant cell washout from the myocardium immediately after balloon deflation. High-concentration, single-bolus therapy may be as effective as balloon-occlusion delivery. This tracking technique should facilitate development of improved delivery strategies for cardiac cell therapy. PMID- 17909261 TI - 18F-FDG PET in detecting primary breast cancer. PMID- 17909262 TI - Identification of regulatory elements in the Cyp19 proximal promoter in rat luteal cells. AB - The cytochrome P450 aromatase (Cyp19) gene encodes an enzyme of crucial importance in the synthesis of estradiol. Estradiol is luteotropic in the rat. In this species, luteal Cyp19 expression increases progressively during pregnancy and falls before parturition. The mechanisms that control these changes are unknown. Using gel shift assays, we sought to identify the promoter regions that control Cyp19 expression in the rat corpus luteum (CL). The Cyp19 promoter contains a cAMP response element-like sequence (CLS), two nuclear receptor elements half sites (NREs), a GATA binding site, a Yin Yang-1 (YY1) response element, and an activation protein 3 (AP3) binding site. Nuclear extracts were obtained from CL of rats on days 4, 15, and 23 of pregnancy and from the ovaries of immature rats treated with vehicle or a hormone that induces Cyp19 expression in the follicles. CLS was active in immature ovaries but inactive in the CL of pregnant rats, whereas binding to NREs and GATA was observed in both tissues. YY1 was inactive in all samples tested. In the CL, AP3 binding was higher on day 15 of pregnancy when compared with day 4 and day 23 but it was absent in ovaries of immature rats, whereas luteinization increased AP3 binding activity. Mutation of the AP3 site blunted the stimulation of Cyp19 promoter activity in granulosa cells. Our results indicate that CLS is active only in follicles; whereas in the CL, binding to the GATA, NRE, and AP3 sites associates with changes in Cyp19 expression, suggesting that they control Cyp19 promoter activity in luteal cells. PMID- 17909263 TI - Distinct expression and activity profiles of largemouth bass (Micropterus salmoides) estrogen receptors in response to estradiol and nonylphenol. AB - The estrogen receptor (ER) signaling cascade is a vulnerable target of exposure to environmental xenoestrogens, like nonylphenol (NP), which are causally associated with impaired health status. However, the impact of xenoestrogens on the individual receptor isotypes (alpha, beta a, and beta b) is not well understood. The goal of these studies was to determine the impact of NP on largemouth bass (Micropterus salmoides) ER isotype expression and activity. Here, we show that hepatic expression levels of three receptors are not equivalent in male largemouth bass exposed to NP by injection. Transcript levels of the ER alpha subtype were predominantly induced in concert with vitellogenin similarly to fish exposed to 17beta-estradiol (E(2)) as measured by quantitative real-time PCR. NP also induced circulating plasma levels of estrogen, which may contribute to overall activation of the ERs. To measure the activation of each receptor isotype by E(2) and NP, we employed reporter assays using an estrogen response element (ERE)-luciferase construct. Results from these studies show that ER alpha had the greatest activity following exposure to E(2) and NP. This activity was inhibited by the antagonists ICI 182 780 and ZM 189 154. Furthermore, both beta b and beta a subtypes depressed ER alpha activation, suggesting that the cellular composition of receptor isotypes may contribute to the overall actions of estrogen and estrogenic contaminants via the receptors. Results from these studies collectively reveal the differential response of fish ER isotypes in response to xenoestrogens. PMID- 17909264 TI - Vitamin K2 induces phosphorylation of protein kinase A and expression of novel target genes in osteoblastic cells. AB - Vitamin K is known as a critical nutrient required for bone homeostasis and blood coagulation, and it is clinically used as a therapeutic agent for osteoporosis in Japan. Besides its enzymatic action as a cofactor of vitamin K-dependent gamma glutamyl carboxylase (GGCX), we have previously shown that vitamin K(2) is a transcriptional regulator of bone marker genes and extracellular matrix-related genes, by activating the steroid and xenobiotic receptor (SXR). To explore a novel action of vitamin K in osteoblastic cells, we identified genes up-regulated by a vitamin K(2) isoform menaquinone-4 (MK-4) using oligonucleotide microarray analysis. Among these up-regulated genes by MK-4, growth differentiation factor 15 (GDF15) and stanniocalcin 2 (STC2) were identified as novel MK-4 target genes independent of GGCX and SXR pathways in human and mouse osteoblastic cells. The induction of GDF15 and STC2 is likely specific to MK-4, as it was not exerted by another vitamin K(2) isoform MK-7, vitamin K(1), or the MK-4 side chain structure geranylgeraniol. Investigation of the involved signaling pathways revealed that MK-4 enhanced the phosphorylation of protein kinase A (PKA), and the MK-4 dependent induction of both GDF15 and STC2 genes was reduced by the treatment with a PKA inhibitor H89 or siRNA against PKA. These results suggest that vitamin K(2) modulates its target gene expression in osteoblastic cells through the PKA dependent mechanism, which may be distinct from the previously known vitamin K signaling pathways. PMID- 17909265 TI - Rho GDP dissociation inhibitor alpha interacts with estrogen receptor alpha and influences estrogen responsiveness. AB - Estrogen receptor alpha (ER alpha) is a ligand-activated transcription factor that regulates expression of estrogen-responsive genes. Upon binding of the ligand-occupied ER alpha to estrogen response elements (EREs) in DNA, the receptor interacts with a variety of coregulatory proteins to modulate transcription of target genes. We have isolated and identified a number of proteins associated with the DNA-bound ER alpha. One of these proteins, Rho guanosine diphosphate (GDP) dissociation inhibitor alpha (RhoGDI alpha), is a negative regulator of the Rho family of GTP-binding proteins. In this study, we demonstrate that endogenously expressed RhoGDI alpha is present in the nucleus as well as the cytoplasm of MCF-7 breast cancer cells, and that RhoGDI alpha binds directly to ER alpha, alters the ER alpha-ERE interaction, and influences the ability of ER alpha to regulate transcription of a heterologous estrogen responsive reporter plasmid in transient transfection assays as well as endogenous, estrogen-responsive genes in MCF-7 cells. Our studies suggest that, in addition to the activity of RhoGDI alpha in the cytoplasm, it also influences ER alpha signaling in the nucleus. PMID- 17909266 TI - Crosstalk of CREB and Fos/Jun on a single cis-element: transcriptional repression of the steroidogenic acute regulatory protein gene. AB - Transcriptional regulation of the steroidogenic acute regulatory (StAR) protein gene by cAMP-dependent mechanisms occurs in the absence of a consensus cAMP response element (CRE; TGACGTCA) and is mediated by several sequence-specific transcription factors. We previously identified three CRE-like sites (within the 151/-1 bp cAMP-responsive region of the mouse StAR gene), of which the CRE2 site overlaps with an activator protein-1 (AP-1) motif (TGACTGA, designated as CRE2/AP 1) that can bind both CRE and AP-1 DNA-binding proteins. The present studies were aimed at exploring the functional crosstalk between CREB (CRE-binding protein) and cFos/cJun (AP-1 family members) on the CRE2/AP-1 element and its role in regulating transcription of the StAR gene. Using MA-10 mouse Leydig tumor cells, we demonstrate that the CRE and AP-1 families of proteins interact with the CRE2/AP-1 sequence. CREB, cFos, and cJun proteins were found to bind to the CRE2/AP-1 motif but not the CRE1 and CRE3 sites. Treatment with the cAMP analog (Bu)(2)cAMP augmented phosphorylation of CREB (Ser(133)), cFos (Thr(325)), and cJun (ser(73)). Chromatin immunoprecipitation studies revealed that the induction of CREB, cFos, and cJun by (Bu)(2)cAMP was correlated with protein-DNA interactions and recruitment of the coactivator CREB-binding protein (CBP) to the StAR promoter. EMSA studies employing CREB and cFos/cJun proteins demonstrated competition between these factors for binding to the CRE2/AP-1 motif. Transfection of cells containing the -151/-1 StAR reporter with CREB and cFos/cJun resulted in trans-repression of the StAR gene, an event tightly associated with CBP, demonstrating that both CREB and Fos/Jun compete with each other for binding with limited amounts of intracellular CBP. Overexpression of adenovirus E1A, which binds and inactivates CBP, markedly suppressed StAR gene expression. Ectopic expression of CBP eliminated the repression of the StAR gene by E1A and potentiated the activity of CREB and cFos/cJun on StAR promoter responsiveness. These findings identify molecular events involved in crosstalk between CREB and cFos/cJun, which confer both gain and loss of function on a single cis-element in fine-tuning of the regulatory events involved in transcription of the StAR gene. PMID- 17909267 TI - The AMP-activated protein kinase activator, 5-aminoimidazole-4-carboxamide-1-b-D ribonucleoside, regulates lactate production in rat Sertoli cells. AB - The aim of the present study was to investigate whether the AMP-activated protein kinase (AMPK), a key regulator of cellular energy homeostasis, is present in Sertoli cells and whether its activation by 5-aminoimidazole-4-carboxamide-1-b-d ribonucleoside (AICAR) results in the regulation of cell metabolism to ensure lactate supply for germ cell development. Sertoli cell cultures from 20-day-old rats were used. Western blot analysis for the alpha-subunit of AMPK showed that high levels of AMPK are present in Sertoli cells. Treatment of the cultures with AICAR resulted in a dose- and time-dependent increase of P-AMPK levels indicating activation of the enzyme. A possible effect of AICAR on Sertoli cell lactate production was then analyzed. A dose- and time-dependent increment in lactate secretion was observed. The participation of AMPK activation in different biochemical processes that may be implicated in the regulation of lactate production was also analyzed. AICAR stimulated glucose uptake in a dose- and time dependent manner. Additionally, AICAR increased the glucose transporter 1 (GLUT1) and decreased the glucose transporter 3 (GLUT3) mRNA levels. As for the role of AMPK in the regulation of the monocarboxylate transporters 1 and 4 (MCT1 and MCT4), it has been observed that AICAR treatment decreased MCT1 and increased MCT4 mRNA levels. In summary, the results presented herein show that AMPK is present in Sertoli cells and that its activation by AICAR increases lactate production as a result, at least in part, of a) an increase in glucose uptake, b) an increase in GLUT1 expression, and c) a decrease in MCT1 and an increase in MCT4 levels. Altogether, these results suggest an important role of AMPK in modulating the nutritional function of Sertoli cells. PMID- 17909268 TI - Role of specificity protein transcription factors in estrogen-induced gene expression in MCF-7 breast cancer cells. AB - Deletion analysis of several 17beta-estradiol (E(2))-responsive genes have identified GC-rich sites that are associated with hormone-induced transactivation in MCF-7 breast cancer cells. However, the role of individual specificity proteins (Sps) in mediating hormone-induced gene expression has not been unequivocally determined. In transient transfection studies using E(2)-responsive GC-rich promoters from the E(2)F1, carbamoylphosphate synthetase/aspartate transcarbamylase/dihydroorotase (CAD), and retinoic acid receptor alpha (RAR alpha) genes, RNA interference using small inhibitory RNAs for Sp1 (iSp1), Sp3 (iSp3), and Sp4 (iSp4) decreased both basal and E(2)-induced transactivation. The contributions of individual Sp proteins to basal and E(2)-induced activity were promoter dependent. iSp1, iSp3, and iSp4 also significantly inhibited hormonal induction of E(2)F1, CAD, and RAR alpha mRNA levels; however, the enhanced inhibitory effects of the latter two small inhibitory RNAs suggest that Sp3 and Sp4 play a major role in estrogen receptor alpha/Sp-mediated gene expression in MCF-7 cells. PMID- 17909269 TI - Oligonucleotide microarray analysis of estrogen receptor alpha-positive postmenopausal breast carcinomas: identification of HRPAP20 and TIMELESS as outstanding candidate markers to predict the response to tamoxifen. AB - The estrogen receptor alpha (ER alpha) status of breast tumors is used to identify patients who may respond to endocrine agents such as tamoxifen. However, ER alpha status alone is not perfectly predictive, and there is a pressing need for more reliable markers of endocrine responsiveness. In this aim, we used a two step strategy. We first screened genes of interest by a pangenomic 44 K oligonucleotide microarray in a series of ten ER alpha-positive tumors from five tamoxifen-treated postmenopausal patients who relapsed (distant metastasis) and five tamoxifen-treated postmenopausal patients who did not relapse, matched with respect to age, Scarff-Bloom-Richardson grade, lymph node status, and macroscopic tumor size. Genes of interest (n=24) were then investigated in an independent well-characterized series of ER alpha-positive unilateral invasive primary breast tumors from postmenopausal women who received tamoxifen alone as adjuvant hormone therapy after primary surgery. We identified four genes (HRPAP20, TIMELESS, PTPLB, and MGC29814) for which high mRNA levels were significantly associated with shorter relapse-free survival (log-rank test). We also showed that hormone regulated proliferation-associated 20 kDa protein (HRPAP20) and TIMELESS are 17beta-estradiol-regulated in vitro and are ectopically expressed in OH-Tam resistant cell lines. In conclusion, these findings point to HRPAP20 and TIMELESS as promising markers of tamoxifen resistance in women with ER alpha-positive breast tumors. PMID- 17909270 TI - 20 beta-hydroxysteroid dehydrogenase and CYP19A1 are differentially expressed during maturation in Atlantic cod (Gadus morhua). AB - In order to better quantify the molecular mechanisms regulating final oocyte maturation and spawning, complete coding sequences with partially or fully untranslated regions for the steroidogenic enzymes, cytochrome P450 aromatase and 20 beta-hydroxysteroid dehydrogenase, were cloned from ovaries of Atlantic cod (Gadus morhua). The nucleotide and amino acid sequences showed high homologies with the corresponding sequences of other fish species, and conserved features important for functionality were identified in both predicted proteins. The sequences of the corresponding genomic loci were also determined, allowing the design of mRNA-specific quantitative PCR assays. As a reference gene for the real time RT-PCR assays, eukaryotic elongation factor 1 alpha was chosen, and the mRNA as well as the genomic sequence was determined. In addition, a real-time quantitative PCR assay for the 18S rRNA was adapted to be used in cod. Analysis of immature and maturing female cod from July to January respectively showed that the enzyme genes showed the expected quantitative changes associated with physiological regulation. However, mRNA for eukaryotic elongation factor 1 alpha, and to a lesser extent even 18S rRNA, showed variable expression in these samples as well. To find accurate standards for real-time PCR in such a dynamic organ as the cod ovary is not an easy task, and several possible solutions are discussed. PMID- 17909271 TI - Emerging pharmacologic options for treating postoperative ileus. AB - PURPOSE: Characteristics of the ideal drug therapy for postoperative ileus (POI); the pharmacology, efficacy, and safety of currently available nonselective opioid antagonists and the new peripherally selective opioid antagonists methylnaltrexone and alvimopan for the treatment of POI; and formulary considerations associated with the introduction of these new POI drug therapies are discussed. SUMMARY: The ideal drug therapy for treating POI would selectively antagonize the inhibitory effects on the gastrointestinal (GI) tract of all of the potential factors implicated in the pathophysiology of POI (neurogenic, inflammatory, hormonal, and pharmacologic mediators). The most promising target to date is inhibition of the adverse GI effects of endogenous and exogenous opioids. Selective inhibition of the mu-opioid receptors in the GI tract, without reversing centrally mediated opioid-induced analgesia, may be beneficial in reducing POI. The nonselective opioid antagonists naloxone and nalmefene have not been studied for POI, and they cross the blood-brain barrier. Therefore, they are not appropriate for preventing or treating POI. The peripherally selective opioid antagonist methylnaltrexone shortens the duration of POI and the hospital length of stay (LOS). Alvimopan, a more extensively studied peripherally selective opioid antagonist, has been shown to reduce the duration of POI, frequency of postoperative nausea and vomiting, and hospital LOS. Both methylnaltrexone and alvimopan also appear effective for treating opioid-induced constipation. Preliminary results of a long-term study of alvimopan safety have revealed some potential concerns, and the significance of the adverse effects must be understood before the most appropriate role of alvimopan in patient care can be determined. Restricting the prescribing of new POI drug therapies to certain types of patients, surgeries, and prescribers; incorporating these therapies into preoperative and postoperative policies, procedures, and protocols; and the potential cost savings from reducing hospital LOS are among the considerations in adding these agents to health-system formularies. CONCLUSION: Peripherally selective opioid receptor antagonists are promising new drug therapies that can reduce the clinical and economic burden of POI. PMID- 17909274 TI - Pathogenesis and clinical and economic consequences of postoperative ileus. AB - PURPOSE: The pathogenesis, etiology, clinical manifestations, and clinical and economic consequences of postoperative ileus (POI) in patients undergoing major abdominal surgery; the estimated prevalence of POI; the potential cost savings from efforts to shorten hospital length of stay (LOS); and the role of patient counseling in minimizing the consequences of POI are discussed. SUMMARY: POI has neurogenic, inflammatory, hormonal, and pharmacologic components. It manifests as abdominal distention, pain, nausea, vomiting, and inability to pass stools or tolerate a solid diet that in half of patients undergoing major abdominal surgery persist for more than four days. Surgical stress and prolonged opioid analgesic use contribute to POI. Delayed surgical wound healing and ambulation, atelectasis, pneumonia, and deep vein thrombosis are among the possible complications of POI that can increase LOS, resource use, and health care costs. POI is common; its prevalence probably is underestimated. The potential cost savings from shortening LOS by one day are substantial. Providing advice about the proper preoperative and postoperative care regimen to patients undergoing major abdominal surgery can minimize the clinical and economic consequences of POI. CONCLUSION: POI is a common complication of major abdominal surgery that can have a substantial clinical and economic impact. PMID- 17909275 TI - Current strategies for preventing or ameliorating postoperative ileus: a multimodal approach. AB - PURPOSE: Internal and external factors that contribute to postoperative ileus (POI), the efficacy and safety of various nonpharmacologic and pharmacologic interventions that have been evaluated for the prevention or amelioration of POI, and the current multimodal approach used in patients undergoing major abdominal surgery are described. SUMMARY: Catecholamine and cytokine release associated with the stress response to surgery and the use of certain antiemetic medications, opioid analgesics, and inhaled anesthetics are among the factors that contribute to POI. Early ambulation does not affect the duration of POI, although it has other benefits for patients undergoing abdominal surgery. Clinical experience supports the use of laparoscopy instead of laparotomy if possible, removal of nasogastric tubes shortly after surgery, restriction of intravenous fluids, and initiation of clear oral liquids and ambulation on the first postoperative day. The recommended therapeutic approach for patients undergoing major abdominal surgery involves thoracic epidural analgesia using a local anesthetic with or without an epidural opioid analgesic, and systemic nonsteroidal anti-inflammatory drugs for their opioid-sparing effect if systemic opioid analgesics are used. Buprenorphine may be preferred if a systemic opioid analgesic is used, because it has little effect on gastrointestinal smooth muscle. Metoclopramide, erythromycin, beta blockers, laxatives, neostigmine, naloxone, and gum chewing are not useful for treating POI. CONCLUSION: Most pharmacologic interventions that have been tried in an effort to prevent or ameliorate POI are ineffective or cause intolerable adverse effects. Research is needed to identify and develop new drug therapies for POI. PMID- 17909277 TI - [Birth asphyxia, neonatal risk factors for hypoxic ischemic encephalopathy]. AB - OBJECTIVE: Neonates suffering from severe birth asphyxia may develop hypoxic ischemic encephalopathy (HIE), some of which develop permanent neurological damage. As the incidence of asphyxia and HIE in Iceland is unknown, this study was conducted. Furthermore, we evaluated the association between some neonatal risk factors and the development of HIE. MATERIAL AND METHODS: All term infants born at LSH from 1997-2001 with birth asphyxia, defined as 5 minute Apgar score or=<6, were included in the study. Clinical information, length and weight, Apgar scores at 1, 5 and 10 minutes normoblasts count, initial pH and hemoglobin levels were retrospectively collected. RESULTS: The incidence of HIE after birth asphyxia was 1.4/1000. The infants who developed HIE had significantly lower birth weight and Apgar scores at one, five and ten minutes. They also had lower umbilical artery pH, had more base deficit and lower serum bicarbonate concentrations than the infants who did not develop HIE. CONCLUSION: The incidence of HIE was low compared to other studies. Birth asphyxia resulting in HIE is associated with lower birth weight, Apgar scores, pH and neonatal hemoglobin levels at birth. We conclude that neonates with low hemoglobin level are at increased risk for developing HIE and that low pH and Apgar scores may predict worse outcomes after birth asphyxia. PMID- 17909278 TI - [Respiratory dysfunction in infants born by elective cesarean section without labor]. AB - OBJECTIVE: To evaluate the effects of gestational age at the timing of elective caesarean section (ECS) on the incidence of respiratory dysfunction in the newborn. STUDY GROUP AND METHODS: This was a retrospective study. All infants born by ECS at the Landspitali-University Hospital Iceland over a 10 years period (1996-2005) at >or=37 weeks gestation and diagnosed with transient tachypnoea of the newborn (TTN) or respiratory distress syndrome (RDS) were included in the study. RESULTS: Of the 1486 infants delivered by ECS over the study period 57 (3.8%) developed TTN (50 infants) or RDS (7 infants). The incidence of respiratory dysfunction was inversely related to gestational age, 13.8% at 37 weeks gestation and 2.5% at 40 weeks gestation. A statistically significant reduction in the incidence of TTN or RDS was observed from 38 weeks to 39 weeks gestation (6.6% and 2.3% respectively; p<0.001). There has been a reduction in the incidence of ECS before 39 weeks gestation since 2001, when guidelines regarding optimal timing of ECS were set at our hospital. CONCLUSION: The incidence of respiratory dysfunction in neonates born by ECS is inversely related to gestational age, even in the term infant. It is important to delay ECS until 39 weeks gestation whenever possible, in order to minimize the risk of respiratory dysfunction in the newborn infant. PMID- 17909279 TI - [Unilateral spatial neglect: a review of symptoms, frequency diagnosis and prognosis]. AB - Unilateral spatial neglect is a disorder commonly encountered after hemisphere stroke, most often in the right hemisphere. Neglect patients fail to attend and respond to stimuli presented on the side of space opposite to the brain lesion. Neglect implies a complex dysfunction in the co-action between perception, motor behavior and the environment where the patients attentive capacities and the environmental space are of special importance. Patients difficulties can be seen in that they do not eat from the left side of the plate or omit words to the left when asked to read. Commonly patients do not have a complete insight into their neglect problems. Neglect in stroke patients has been associated with poor outcome on functional activities. Signs of neglect are not always obvious but can be explored and assessed quickly by bedside neuropsychological testing. Neglect is often more unclear to an observer than, e.g. if a patient suffers from paresis or aphasia. Education for patients, their relatives and others are therefore important. PMID- 17909280 TI - Structure of Alzheimer's disease amyloid precursor protein copper-binding domain at atomic resolution. AB - Amyloid precursor protein (APP) plays a central role in the pathogenesis of Alzheimer's disease, as its cleavage generates the Abeta peptide that is toxic to cells. APP is able to bind Cu2+ and reduce it to Cu+ through its copper-binding domain (CuBD). The interaction between Cu2+ and APP leads to a decrease in Abeta production and to alleviation of the symptoms of the disease in mouse models. Structural studies of CuBD have been undertaken in order to better understand the mechanism behind the process. Here, the crystal structure of CuBD in the metal free form determined to ultrahigh resolution (0.85 A) is reported. The structure shows that the copper-binding residues of CuBD are rather rigid but that Met170, which is thought to be the electron source for Cu2+ reduction, adopts two different side-chain conformations. These observations shed light on the copper binding and redox mechanisms of CuBD. The structure of CuBD at atomic resolution provides an accurate framework for structure-based design of molecules that will deplete Abeta production. PMID- 17909281 TI - Structure of 3(17)alpha-hydroxysteroid dehydrogenase (AKR1C21) holoenzyme from an orthorhombic crystal form: an insight into the bifunctionality of the enzyme. AB - Mouse 3(17)alpha-hydroxysteroid dehydrogenase (AKR1C21) is a bifunctional enzyme that catalyses the oxidoreduction of the 3- and 17-hydroxy/keto groups of steroid substrates such as oestrogens, androgens and neurosteroids. The structure of the AKR1C21-NADPH binary complex was determined from an orthorhombic crystal belonging to space group P2(1)2(1)2(1) at a resolution of 1.8 A. In order to identify the factors responsible for the bifunctionality of AKR1C21, three steroid substrates including a 17-keto steroid, a 3-keto steroid and a 3alpha hydroxysteroid were docked into the substrate-binding cavity. Models of the enzyme-coenzyme-substrate complexes suggest that Lys31, Gly225 and Gly226 are important for ligand recognition and orientation in the active site. PMID- 17909282 TI - Structure of Mycobacterium tuberculosis mtFabD, a malonyl-CoA:acyl carrier protein transacylase (MCAT). AB - Mycobacteria display a unique and unusual cell-wall architecture, central to which is the membrane-proximal mycolyl-arabinogalactan-peptidoglycan core (mAGP). The biosynthesis of mycolic acids, which form the outermost layer of the mAGP core, involves malonyl-CoA:acyl carrier protein transacylase (MCAT). This essential enzyme catalyses the transfer of malonyl from coenzyme A to acyl carrier protein AcpM, thus feeding these two-carbon units into the chain elongation cycle of the type II fatty-acid synthase. The crystal structure of M. tuberculosis mtFabD, the mycobacterial MCAT, has been determined to 3.0 A resolution by multi-wavelength anomalous dispersion. Phasing was facilitated by Ni2+ ions bound to the 20-residue N-terminal affinity tag, which packed between the two independent copies of mtFabD. PMID- 17909284 TI - Cocrystallizing natural RNA with its unnatural mirror image: biochemical and preliminary X-ray diffraction analysis of a 5S rRNA A-helix racemate. AB - Chemically synthesized RNAs with the unnatural L-configuration possess enhanced in vivo stability and nuclease resistance, which is a highly desirable property for pharmacological applications. For a structural comparison, both L- and D-RNA oligonucleotides of a shortened Thermus flavus 5S rRNA A-helix were chemically synthesized. The enantiomeric RNA duplexes were stochiometrically cocrystallized as a racemate, which enabled analysis of the D- and L-RNA enantiomers in the same crystals. In addition to a biochemical investigation, diffraction data were collected to 3.0 A resolution using synchrotron radiation. The crystals belonged to space group P3(1)21, with unit-cell parameters a = b = 35.59, c = 135.30 A, gamma = 120 degrees and two molecules per asymmetric unit. PMID- 17909283 TI - Purification, crystallization, X-ray diffraction analysis and phasing of an engineered single-chain PvuII restriction endonuclease. AB - The restriction endonuclease PvuII from Proteus vulgaris has been converted from its wild-type homodimeric form into the enzymatically active single-chain variant scPvuII by tandemly joining the two subunits through the peptide linker Gly-Ser Gly-Gly. scPvuII, which is suitable for the development of programmed restriction endonucleases for highly specific DNA cleavage, was purified and crystallized. The crystals diffract to a resolution of 2.35 A and belong to space group P4(2), with unit-cell parameters a = b = 101.92, c = 100.28 A and two molecules per asymmetric unit. Phasing was successfully performed by molecular replacement. PMID- 17909285 TI - Crystallographic characterization of the radixin FERM domain bound to the cytoplasmic tail of adhesion molecule CD44. AB - CD44 is an important adhesion molecule that specifically binds hyaluronic acid and regulates cell-cell and cell-matrix interactions. Increasing evidence has indicated that CD44 is assembled in a regulated manner into the membrane cytoskeletal junction, a process that is mediated by ERM (ezrin/radixin/moesin) proteins. Crystals of a complex between the radixin FERM domain and the C terminal cytoplasmic region of CD44 have been obtained. The crystal of the radixin FERM domain bound to the CD44 cytoplasmic tail peptide belongs to space group P2(1)2(1)2(1), with unit-cell parameters a = 62.70, b = 66.18, c = 86.22 A, and contain one complex in the crystallographic asymmetric unit. An intensity data set was collected to a resolution of 2.1 A. PMID- 17909286 TI - Purification, crystallization and initial crystallographic characterization of peanut major allergen Ara h 3. AB - The peanut is a significant food source, but is responsible for many cases of anaphylaxis. The peanut 11S legumin-like seed storage protein Ara h 3 is one of the best characterized allergens. In this study, Ara h 3 was extracted from peanut kernels and purified by sequential anion-exchange, hydrophobic interaction and gel-filtration chromatography to very high purity to facilitate crystallization and structural studies. Well diffracting single crystals were obtained by the vapor-diffusion method. A molecular-replacement structural solution has been obtained and refinement of the structure is currently under way. PMID- 17909287 TI - Isolation, crystallization and preliminary crystallographic analysis of Salmonella typhimurium uridine phosphorylase crystallized with 2,2' anhydrouridine. AB - Uridine phosphorylase (UPh; EC 2.4.2.3) is a member of the pyrimidine nucleoside phosphorylase family of enzymes which catalyzes the phosphorolytic cleavage of the C-N glycoside bond of uridine, with the formation of ribose 1-phosphate and uracil. This enzyme has been shown to be important in the activation and catabolism of fluoropyrimidines. Modulation of its enzymatic activity may affect the therapeutic efficacy of chemotherapeutic agents. The structural investigation of the bacterial uridine phosphorylases, both unliganded and complexed with substrate/product analogues and inhibitors, may help in understanding the catalytic mechanism of the phosphorolytic cleavage of uridine. Salmonella typhimurium uridine phosphorylase has been crystallized with 2,2'-anhydrouridine. X-ray diffraction data were collected to 2.15 A. Preliminary analysis of the diffraction data indicates that the crystal belongs to space group P2(1)2(1)2(1), with unit-cell parameters a = 88.52, b = 123.98, c = 133.52 A. The solvent content is 45.51%, assuming the presence of one hexamer molecule per asymmetric unit. PMID- 17909288 TI - Crystallization and preliminary crystallographic analysis of the ferredoxin component of carbazole 1,9a-dioxygenase from Nocardioides aromaticivorans IC177. AB - Carbazole 1,9a-dioxygenase (CARDO) catalyzes the dihydroxylation of carbazole by angular position (C9a) carbon bonding to the imino nitrogen and its adjacent C1 carbon. CARDO consists of a terminal oxygenase component and two electron transfer components: ferredoxin and ferredoxin reductase. The ferredoxin component of carbazole 1,9a-dioxygenase from Nocardioides aromaticivorans IC177 was crystallized at 293 K using the hanging-drop vapour-diffusion method with ammonium sulfate as the precipitant. The crystals, which were improved by macroseeding, diffract to 2.0 A resolution and belong to space group P4(1)2(1)2. PMID- 17909289 TI - Human tRNA(Gly) acceptor-stem microhelix: crystallization and preliminary X-ray diffraction analysis at 1.2 A resolution. AB - The major dissimilarities between the eukaryotic/archaebacterial-type and eubacterial-type glycyl-tRNA synthetase systems (GlyRS; class II aminoacyl-tRNA synthetases) represent an intriguing example of evolutionarily divergent solutions to similar biological functions. The differences in the identity elements of the respective tRNA(Gly) systems are located within the acceptor stem and include the discriminator base U73. In the present work, the human tRNA(Gly) acceptor-stem microhelix was crystallized in an attempt to analyze the structural features that govern the correct recognition of tRNA(Gly) by the eukaryotic/archaebacterial-type glycyl-tRNA synthetase. The crystals of the human tRNA(Gly) acceptor-stem helix belong to the monoclinic space group C2, with unit cell parameters a = 37.12, b = 37.49, c = 30.38 A, alpha = gamma = 90, beta = 113.02 degrees, and contain one molecule per asymmetric unit. A high-resolution data set was acquired using synchrotron radiation and the data were processed to 1.2 A resolution. PMID- 17909290 TI - Crystallization and preliminary X-ray characterization of arylamine N acetyltransferase C (BanatC) from Bacillus anthracis. AB - The arylamine N-acetyltransferase (NAT) enzymes are xenobiotic metabolizing enzymes that have been found in a large range of eukaryotes and prokaryotes. These enzymes catalyse the acetylation of arylamine drugs and/or pollutants. Recently, a Bacillus anthracis NAT isoform (BanatC) has been cloned and shown to acetylate the sulfonamide antimicrobial sulfamethoxazole (SMX). Subsequently, it was shown that BanatC contributes to the resistance of this bacterium to SMX. Here, the crystallization and the X-ray characterization of BanatC (Y38F mutant) are reported. The crystals belong to the tetragonal space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 53.70, c = 172.40 A, and diffract to 1.95 A resolution on a synchrotron source. PMID- 17909291 TI - Crystallization and preliminary X-ray diffraction of human interleukin-7 bound to unglycosylated and glycosylated forms of its alpha-receptor. AB - The interleukin-7 (IL-7) signaling pathway plays an essential role in the development, proliferation and homeostasis of T and B cells in cell-mediated immunity. Understimulation and overstimulation of the IL-7 signaling pathway leads to severe combined immunodeficiency, autoimmune reactions, heart disease and cancers. Stimulation of the IL-7 pathway begins with IL-7 binding to its alpha-receptor, IL-7R alpha. Protein crystals of unglycosylated and glycosylated complexes of human IL-7-IL-7R alpha extracellular domain (ECD) obtained using a surface entropy-reduction approach diffract to 2.7 and 3.0 A, respectively. Anomalous dispersion methods will be used to solve the unglycosylated IL-7-IL-7R alpha ECD complex structure and this unglycosylated structure will then serve as a model in molecular-replacement attempts to solve the structure of the glycosylated IL-7-alpha-receptor complex. PMID- 17909292 TI - Expression, purification, crystallization and preliminary X-ray crystallographic analysis of a resuscitation-promoting factor from Mycobacterium tuberculosis. AB - The resuscitation-promoting factor RpfB, the most complex of the five resuscitation-promoting factors produced by M. tuberculosis, is devoted to bacterial reactivation from the dormant state. RpfB consists of 362 residues predicted to form five domains. An RpfB fragment containing the protein catalytic domain and a G5 domain has been successfully crystallized using vapour-diffusion methods. This is the first crystallographic study of a resuscitation-promoting factor. Crystals of this protein belong to space group I422, with unit-cell parameters a = 97.63, b = 97.63, c = 114.87 A. Diffraction data have also been collected from a selenomethionine derivative at 2.9 A resolution. Model building using the phases derived from the multiwavelength anomalous dispersion experiment is in progress. PMID- 17909293 TI - Crystallization and preliminary X-ray data analysis of beta-alanine synthase from Drosophila melanogaster. AB - Beta-alanine synthase catalyzes the last step in the reductive degradation pathway for uracil and thymine, which represents the main clearance route for the widely used anticancer drug 5-fluorouracil. Crystals of the recombinant enzyme from Drosophila melanogaster, which is closely related to the human enzyme, were obtained by the hanging-drop vapour-diffusion method. They diffracted to 3.3 A at a synchrotron-radiation source, belong to space group C2 (unit-cell parameters a = 278.9, b = 95.0, c = 199.3 A, beta = 125.8 degrees) and contain 8-10 molecules per asymmetric unit. PMID- 17909294 TI - Purification, crystallization and preliminary structural characterization of the N-terminal region of the human formin-homology protein FHOD1. AB - Formins are key regulators of actin cytoskeletal dynamics that constitute a diverse protein family that is present in all eukaryotes examined. They typically consist of more than 1000 amino acids and are defined by the presence of two conserved regions, namely the formin homology 1 and 2 domains. Additional conserved domains comprise a GTPase-binding domain for activation, a C-terminal autoregulation motif and an N-terminal recognition domain. In this study, the N terminal region (residues 1-339) of the human formin homology domain-containing protein 1 (FHOD1) was purified and crystallized from 20%(w/v) PEG 4000, 10%(v/v) glycerol, 0.3 M magnesium chloride and 0.1 M Tris-HCl pH 8.0. Native crystals belong to space group P1, with unit-cell parameters a = 35.4, b = 73.9, c = 78.7 A, alpha = 78.2, beta = 86.2, gamma = 89.7 degrees. They contain two monomers of FHOD1 in the asymmetric unit and diffract to a resolution of 2.3 A using a synchrotron-radiation source. PMID- 17909295 TI - Protein preparation, crystallization and preliminary X-ray crystallographic analysis of SMU.961 protein from the caries pathogen Streptococcus mutans. AB - The smu.961 gene encodes a putative protein of 183 residues in Streptococcus mutans, a major pathogen in human dental caries. The gene was cloned into expression vector pET28a and expressed in a substantial quantity in Escherichia coli strain BL21 (DE3) with a His tag at its N-terminus. The recombinant protein SMU.961 was purified to homogeneity in a two-step procedure consisting of Ni2+ chelating and size-exclusion chromatography. Crystals suitable for X-ray diffraction were obtained by the hanging-drop vapour-diffusion method and diffracted to 2.9 A resolution at beamline I911-3, MAX-II-lab, Sweden. The crystal belonged to space group C2, with unit-cell parameters a = 98.62, b = 73.73, c = 184.73 A, beta = 98.82 degrees. PMID- 17909296 TI - Overproduction, purification and preliminary X-ray diffraction analysis of CzcE from Cupriavidus metallidurans CH34. AB - CzcE is encoded by the czc determinant that allows Cupriavidus metallidurans CH34 to modulate its internal concentrations of cobalt, zinc and cadmium. This periplasmic protein was overproduced in its mature form in Escherichia coli and purified in two steps. After preliminary screening of crystallization conditions using a robot, well diffracting crystals were obtained using the hanging-drop vapour-diffusion method. Crystals diffracted to 1.96 A using synchrotron radiation. They belonged to the monoclinic space group C2, with unit-cell parameters a = 105.54, b = 29.68, c = 71.10 A. The asymmetric unit is expected to contain a dimer, in agreement with the quaternary structure deduced from gel filtration experiments. PMID- 17909297 TI - Crystallization of sheep (Ovis aries) and goat (Capra hircus) haemoglobins under unbuffered low-salt conditions. AB - Haemoglobin is a tetrameric protein that plays a vital role in the transport of oxygen from the lungs to the tissues and of carbon dioxide back to the lungs. Even though a large amount of work has already been performed in this area, the study of the haemoglobin structures of avian and mammalian species is rather incomplete. Efforts are being made to understand the salient features of the species mentioned above. Here, whole blood plasma was collected from sheep and goat and purified by anion-exchange chromatography; the haemoglobins were crystallized by the hanging-drop vapour-diffusion method under unbuffered low salt conditions using PEG 3350 as a precipitant. Data collection was carried out using a MAR345 image-plate detector system. Sheep haemoglobin crystallizes in the orthorhombic space group P2(1)2(1)2(1) with one whole biological molecule (alpha2beta2) in the asymmetric unit, with unit-cell parameters a = 60.231, b = 70.695, c = 131.479 A. In contrast, goat haemoglobin crystallizes in the triclinic system with two biological molecules (alpha2beta2) in the unit cell. The unit-cell parameters are a = 53.103, b = 69.382, c = 96.098 A, alpha = 110.867, beta = 91.133, gamma = 109.437 degrees. PMID- 17909298 TI - Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DsrEFH from Allochromatium vinosum. AB - In purple sulfur bacteria, the proteins encoded by dsr genes play an essential role in the oxidation of intracellular sulfur, which is an obligate intermediate during the oxidation of sulfide and thiosulfate. One such gene product, DsrEFH from Allochromatium vinosum, has been cloned, expressed, purified and crystallized. Synchrotron data were collected to 2.5 A from a crystal of selenomethionine-substituted DsrEFH. The crystal belongs to the primitive monoclinic space group P2(1), with unit-cell parameters a = 56.6, b = 183.1, c = 107.8 A, beta = 99.6 degrees. A full structure determination is under way in order to provide insight into the structure-function relationships of this protein. PMID- 17909299 TI - Crystallization and preliminary crystallographic analysis of the fourth FAS1 domain of human BigH3. AB - The protein BigH3 is a cell-adhesion molecule induced by transforming growth factor-beta (TGF-beta). It consists of four homologous repeat domains known as FAS1 domains; mutations in these domains have been linked to corneal dystrophy. The fourth FAS1 domain was expressed in Escherichia coli B834 (DE3) (a methionine auxotroph) and purified by DEAE anion-exchange and gel-filtration chromatography. The FAS1 domain was crystallized using the vapour-diffusion method. A SAD diffraction data set was collected to a resolution of 2.5 A at 100 K. The crystal belonged to space group P6(1) or P6(5) and had two molecules per asymmetric unit, with unit-cell parameters a = b = 62.93, c = 143.27 A, alpha = beta = 90.0, gamma = 120.0 degrees. PMID- 17909302 TI - Gambling and increased sexual desire with dopaminergic medications in restless legs syndrome. AB - OBJECTIVES: Do patients with restless legs syndrome (RLS) report gambling or other abnormal behaviors as previously reported in Parkinson disease. METHODS: This survey study was sent to 261 idiopathic RLS patients, and it included the Gambling Symptoms Assessment Scale, Altman Self-Rating Mania Scale, and questions pertaining to sexual activity and novelty-seeking behaviors. RESULTS: Ninety-nine patients responded to the survey, and 77 were actively taking 1 or more dopaminergic medications. Of the 70 respondents who answered the gambling questions, 5 (7%) noted a change in gambling, with 4 (6%; 95% confidence interval, 2%-14%) stating that increased urges and time spent gambling occurred specifically after the use of dopaminergic medications (2 on pramipexole, 1 on ropinirole, and 1 on levodopa and pramipexole). Increased sexual desire was reported by 4 (5%) of the 77 respondents, 3 (4%; 95% confidence interval, 1%-11%) reported that this occurred specifically after the use of dopaminergic medications (1 on pramipexole, 1 on ropinirole, and 1 on levodopa). One patient reported both an increase in gambling and sexual habits. CONCLUSIONS: This exploratory survey study revealed the development of gambling and/or increased sexuality in patients with RLS. These data raise the possibility that, as in Parkinson disease, RLS patients should be cautioned about potential behaviors that may occur with the use of dopaminergic medications. Further prospective studies are needed to assess the relationship between these medications and compulsive behaviors associated with the treatment of RLS. PMID- 17909300 TI - Preparation, crystallization and preliminary crystallographic analysis of old yellow enzyme from Trypanosoma cruzi. AB - Old yellow enzyme (OYE) is an NADPH oxidoreductase that contains a flavin mononucleotide as a prosthetic group. The OYE from Trypanosoma cruzi, which produces prostaglandin F(2alpha), a potent mediator of various physiological and pathological processes, from prostaglandin H2. The protein was recombinantly expressed and purified from Escherichia coli and was crystallized using the hanging-drop vapour-diffusion method. The crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 56.3, b = 78.8, c = 78.8 A, beta = 93.4 degrees and two molecules per asymmetric unit. The crystals were suitable for X-ray crystallographic studies and diffracted to 1.70 A resolution. A Patterson search method is in progress using the structure of OYE from Pseudomonas putida as a starting model. PMID- 17909303 TI - Overnight switch from oral dopaminergic agonists to transdermal rotigotine patch in subjects with Parkinson disease. AB - OBJECTIVE: To assess safety, tolerability, and efficacy outcomes of an overnight switch from oral ropinirole, pramipexole, or cabergoline to rotigotine, a dopaminergic agonist with transdermal delivery over 24 hours in subjects with established Parkinson disease (PD). METHODS: In this open-label multicenter study, we hypothesized that the selected doses of transdermal rotigotine would provide at least equivalent antiparkinsonian actions in subjects with idiopathic PD not adequately controlled with oral ropinirole (up to 9 mg/d), pramipexole (up to 2 mg/d), or cabergoline (up to 3 mg/d). The tolerability of the rotigotine switch was evaluated by the number of subjects completing the scheduled 28-day treatment period, need for rotigotine dose reductions, and dropouts due to adverse events. Efficacy assessment relied on changes in Unified Parkinson's Disease Rating Scale from the baseline to the end of treatment in PD symptoms and subject preference of dopaminergic agonist. RESULTS: Of 116 PD subjects enrolled, 104 completed the 28-day rotigotine treatment. Fifteen subjects required rotigotine dose adjustment; of these, 11 completed the trial. The most common adverse events (generally mild or moderate in intensity) were application site reactions, nausea, and somnolence. The change to rotigotine was well tolerated. Rotigotine was preferred by 77% of subjects who were not adequately controlled by their previous oral dopaminergic agonist. The predetermined rotigotine substitutions provided improvements over baseline in Unified Parkinson's Disease Rating Scale II and III subscales. CONCLUSIONS: Subjects and clinicians found the overnight switch to rotigotine convenient, well tolerated, and effective for control of PD signs and symptoms for subjects previously receiving low-to moderate doses of oral dopaminergic agonists. PMID- 17909304 TI - The Patient Card questionnaire to identify wearing-off in Parkinson disease. AB - This study sought to evaluate the feasibility and performance of the Patient Card (PC), an instrument derived from the Patient Questionnaire by Stacy et al, for detection of wearing-off (W-O), in a very first application to a Spanish population with Parkinson disease. Patients were classified as experiencing W-O on the basis of the clinician's Follow-up Questionnaire, the Unified Parkinson Disease Rating Scale-Item 36, or improvement, after the following scheduled dose of medication, of 1 or more symptoms included in the PC. The highest proportion of patients with W-O was identified by the PC (79.8%), followed by the Follow-up Questionnaire (55.4%), and Unified Parkinson Disease Rating Scale (35.1%) (Cochran Q, P < 0.01). The PC Spanish version performed adequately, capturing W-O symptoms more frequently than did the other methods, as in the original Patient Questionnaire study by Stacy et al. Accordingly, the PC may potentially help clinicians identify W-O phenomena and thereby allow for better management of patients with Parkinson disease. PMID- 17909305 TI - Hormonal replacement therapy in women with Parkinson disease and levodopa-induced dyskinesia: a crossover trial. AB - Eleven postmenopausal women with Parkinson disease and levodopa-induced peak-dose dyskinesias underwent a double-blind, placebo-controlled, crossover study. The active treatment consisted of estrogen replacement therapy for 12 weeks, followed by medroxyprogesterone acetate for 2 weeks. Estrogen replacement therapy medroxyprogesterone acetate administration significantly improved peak-dose dyskinesia without worsening motor disability, thus suggesting a possible benefit on dyskinesias in postmenopausal women with Parkinson disease. PMID- 17909306 TI - Results from a 2-year centralized tolcapone liver enzyme monitoring program. AB - OBJECTIVES: Tolcapone, administered with levodopa/carbidopa to patients with Parkinson disease, is an effective and generally well-tolerated adjunctive therapy. However, 4 early cases of hepatotoxicity causing 3 deaths in patients not properly monitored gave rise to more rigorous liver function test monitoring guidelines and a liver function test monitoring program in which blood samples from tolcapone-treated patients were tested and results were collected in a central database. We analyzed these results to determine the percentages of patients with at least 1 aspartate aminotransferase (AST) or alanine aminotransferase (ALT) elevation above the upper limit of normal (ULN) and at least 1 elevation greater than 2 times the ULN. METHODS: This retrospective, observational analysis included all AST and ALT values recorded in the database from 11,883 samples from patients who received tolcapone for up to 2 years or longer (January 1999-January 2001). RESULTS: Of 1725 patients who had at least 1 AST or ALT measurement, 3.9% (n = 67) had elevations above the ULN. Less than 1% (15/1725) of patients had AST or ALT elevations greater than 2 times the ULN. Most values returned to normal during continued tolcapone treatment. In 472 patients monitored biweekly while receiving tolcapone for 20 to 114 consecutive weeks, only 0.6% (n = 3) had an AST or ALT value greater than 2 times the ULN; no significant elevations occurred in the remaining 469 (99.4%). CONCLUSIONS: This analysis of results from the central monitoring program suggests that significant transaminase elevations are rare, typically transient, and may return to normal in the face of continued tolcapone therapy in most patients. PMID- 17909307 TI - Tolcapone: an efficacy and safety review (2007). AB - Tolcapone (Tasmar), an inhibitor of catechol-O-methyltransferase, is an effective antiparkinsonian agent when used as an adjunct to levodopa in patients with Parkinson disease who have end-of-dose motor fluctuations. In clinical trials, tolcapone significantly reduced "off" time and levodopa requirements. The drug is generally well tolerated, with the most common adverse events being dopaminergic related. However, clinical trials demonstrated dose-related increases in liver enzymes, and postmarketing surveillance noted 4 cases of acute hepatotoxicity with 3 fatalities that were attributed to tolcapone. For this reason, the drug was withdrawn from the market in some countries, and its use was severely restricted in the United States. An analysis of safety data indicates that, since the labeling restrictions in 1998, there have been more than 40,000 patient-years of tolcapone treatment worldwide, with only 3 reports of severe, but reversible, liver injury and no reports of hepatic fatality. It can be concluded that severe liver injury due to tolcapone is a rare event. Based on these data, the drug has been reintroduced to the market in several European countries, and the Food and Drug Administration in the United States has modified monitoring requirements. The new labeling recommends monitoring of liver function every 2 to 4 weeks for 6 months and at the physician's discretion thereafter. In addition, patients must be taken off the drug if blood tests show enzyme elevation of greater than twice the upper limit of normal. This article reviews the data pertaining to the safety and efficacy of tolcapone. PMID- 17909308 TI - Selegiline orally disintegrating tablets in patients with Parkinson disease and "wearing off" symptoms. AB - BACKGROUND: Selegiline orally disintegrating tablet (ODT; Zelapar) is a selective monoamine oxidase B inhibitor developed as an adjunct to levodopa (LD) for Parkinson disease. Most patients on long-term LD therapy eventually experience deterioration at the end of the LD dosing interval, with predictable "wearing off" and "on-off" fluctuations. METHODS: We conducted a 12-week, double-blind, placebo-controlled, parallel-design trial of selegiline ODT. The primary efficacy point was reduction in the percentage of average daily "off" time. Secondary measures included reductions in daily off hours and total daily off time, Clinical Global Impressions-Improvement (CGI-I), and Patient Global Impression Improvement (PGI-I). Patients on LD received selegiline ODT (1.25 mg/d for 6 weeks, then 2.5 mg/d for 6 weeks) or placebo. Safety and tolerability were measured. RESULTS: The intent-to-treat population included 98 patients receiving selegiline ODT and 50 patients receiving placebo. Combined efficacy results for weeks 10 and 12 revealed an 11.6% reduction in percentage of daily off time for selegiline ODT versus a 9.8% reduction for placebo (NS). PGI-I detected a statistically significant difference between treatment groups in favor of selegiline ODT (P = 0.02), whereas CGI-I detected a strong trend toward improvement (P = 0.06). Selegiline ODT was safe and well tolerated. CONCLUSIONS: This study showed no significant difference in improvement in percentage of off time with selegiline ODT versus placebo. Some clinical impressions (e.g., PGI-I, CGI-I) improved. This result contrasts with an identically designed study that showed a significant improvement in off time with selegiline ODT. A combined analysis of both studies suggested overall efficacy. PMID- 17909309 TI - Placebo response in Parkinson trials using patient diaries: sites do matter. PMID- 17909310 TI - A perspective on adjunctive therapy for Parkinson disease with monoamine oxidase B inhibition. PMID- 17909311 TI - Topiramate as treatment for hemifacial spasm. AB - OBJECTIVE: To report a patient with hemifacial spasm (HFS) which improved with topiramate. CASE REPORT: A 52-year-old woman diagnosed with migraine, who also had a right HFS, did experience a marked improvement of HFS after introduction of topiramate as prophylactic therapy for her migraine episodes. Topiramate withdrawal led to reappearance of HFS, which improved again after its reintroduction. CONCLUSIONS: Topiramate should be considered as a possible therapy for HFS. PMID- 17909312 TI - Iatrogenic botulism due to therapeutic botulinum toxin a injection in a pediatric patient. AB - Botulinum toxin A is commonly used to reduce spasticity and dystonia in children with cerebral palsy. We report a pediatric patient who developed systemic botulism as a result of a severe overdose of the injected toxin (40 U/kg). This case highlights the importance of physicians having adequate knowledge of primate and human literature on the lethal dose, 50% of botulinum toxin A before injecting children. PMID- 17909313 TI - Dapoxetine in the treatment of premature ejaculation. PMID- 17909314 TI - Dapoxetine and paroxetine for the treatment of premature ejaculation. PMID- 17909315 TI - A phase 1/2 comparative vaccine trial of the safety and immunogenicity of a CRF01_AE (subtype E) candidate vaccine: ALVAC-HIV (vCP1521) prime with oligomeric gp160 (92TH023/LAI-DID) or bivalent gp120 (CM235/SF2) boost. AB - BACKGROUND: The development of an effective HIV-1 vaccine is critical to control the pandemic. A prime-boost HIV-1 vaccine trial assessing safety and immunogenicity was conducted in Thailand as part of an evaluation of candidate regimens for a phase 3 efficacy trial. METHODS: ALVAC-HIV (vCP1521), expressing circulating recombinant form 01_AE (CRF01_AE) gp120/subtype B LAI and subtype B Gag/Protease boosted with recombinant envelope oligomeric CRF01_AE gp160 (ogp160) or bivalent CRF01_AE/subtype B gp120 CM235/SF2, was evaluated in a phase 1/II trial of 130 HIV-negative Thai adults. RESULTS: One hundred forty volunteers were enrolled, and 130 completed all safety and immunogenicity visits. Reactogenicity was common but generally mild, and there was no significant difference in the adverse event rate between vaccine and placebo recipients (P = 0.26). There were 7 serious adverse events during the follow-up period, none of which were vaccine related. Cumulative HIV-specific, CD8-mediated, cytotoxic T-lymphocyte responses were observed in 11 (25%) of 44 subjects who received ALVAC boosted by bivalent gp120 and in 5 (11%) of 45 subjects who received ALVAC boosted by ogp160, but these differences were not statistically significant compared with those in placebo recipients (P = 0.62 and P = 0.37, respectively). HIV-specific lymphoproliferative responses were detected in 84% of subunit-boosted vaccine recipients and in 10% of placebo recipients. Neutralizing antibody responses to CRF01_AE and subtype B laboratory strains were seen in 95% of ogp160-boosted and 100% of gp120 B/E-boosted vaccinees, respectively. CONCLUSIONS: These 2 different prime-boost regimens seem to be safe and displayed cell-mediated immune responses consistent with those in other trials of canarypox vectors. PMID- 17909317 TI - Early virologic rebound in a pilot trial of ritonavir-boosted atanazavir as maintenance monotherapy. PMID- 17909318 TI - Opportunities and options for the use of protease inhibitors in a resource-poor setting: experiences from southern India. PMID- 17909319 TI - Optimum time to initiate antiretroviral therapy in patients with HIV-associated tuberculosis: there may be more than one right answer. PMID- 17909321 TI - A model of caring mentorship for nursing. AB - Mentoring is a multidimensional relationship that energizes personal and professional growth. This article explores the concept of mentoring in nursing and presents a mentorship model based on a caring philosophy. The RN-student nurse mentoring program cited is the result of a collaborative commitment between a community hospital and two colleges. Discussed are the experience, process, insights, and impact of the program as a retention and professional development tool. PMID- 17909323 TI - Experienced registered nurses' satisfaction with using self-learning modules versus traditional lecture/discussion to achieve competency goals during hospital orientation. AB - Staff development educators are challenged with various levels of experience and learning styles among newly hired registered nurses in a hospital orientation group. This diversity forces the educator to use various teaching strategies to meet the learning needs of the group. This study investigated the experienced registered nurses' satisfaction with using self-learning modules versus traditional lecture/discussion methods during the nursing orientation process. The results revealed that experienced nurses prefer the more traditional method of learning by lecture/discussion. PMID- 17909325 TI - Video as a strategy to evaluate and assist clinical teachers with student assessment. AB - The assessment of the clinical competence of nursing students requires expertise in nursing as well as skills in student assessment. The development of these student assessment skills is dependent on the preparation of the clinical teacher. The results of this pilot study indicate that video can be one successful tool in evaluating the assessment skills of clinical teachers. Video also may be useful as a staff development tool by providing a nonthreatening way for clinical teachers to familiarize themselves with clinical assessment. In turn, this will have benefits for universities and students by ensuring consistency of clinical assessment by clinical teachers. PMID- 17909326 TI - The effect of problem-solving training on the counseling skills of telephonic nurse care managers. AB - Nurses increasingly use telephonic assessment and counseling to manage clients with a variety of chronic illnesses. This article describes a study designed to assess a group of telephonic nurse disease managers' teaching and adherence promotion skills during actual patient interactions. Nurse care managers showed improvements after training in four main counseling skills categories, with a decrease in time spent. PMID- 17909328 TI - Guiding the nurse educator: advice to mentors. PMID- 17909329 TI - Preceptorship. PMID- 17909327 TI - The One-Minute Preceptor: a five-step tool to improve clinical teaching skills. AB - The One-Minute Preceptor (OMP) is a teaching tool that has been used successfully for over 10 years in family practice residency programs. It was designed to enhance the teaching skills of physicians involved in the clinical education of new residents. This article describes the five steps of the OMP and how it was taught to a group of nurse preceptors and reports their evaluations of the impact that this education had on their ability to instruct and offer feedback to the novice nurse. PMID- 17909330 TI - Instant teaching tools. PMID- 17909334 TI - Normal reference values of Southwick's anteroposterior angle in prepubertal and pubertal normal adolescents. AB - Measurement of the Southwick's anteroposterior (AP) angle (shaft epiphysis proximal femoral AP angle) is not only a useful tool for planning the surgical treatment of deformities caused by slipped capital femoral epiphysis, but seems to be also important for recognizing the risk of epiphysiolysis development in obese patients (increased AP angle) or to confirm the diagnosis of slipped capital femoral epiphysis (decreased AP angle). To establish normal reference values of the Southwick's AP angle, we studied 97 normal nonobese adolescents (42 females, 55 males), with ages ranging between 8 and 16 years. The mean (SD) AP angle was 151.2 (5.0), ranging from 140 to 164. The limits for the first (p25) and third (p75) quartiles were 148 and 155, respectively. No difference was observed in the AP angle in males when compared with females. The AP angle was evaluated according to sex, chronological age, bone age, weight, height, and pubertal stage of development. We observed an inverse correlation of the AP angle with chronological age (r=-0.57) and bone age (r=-0.52). A weak inverse correlation was also found with stature (r=-0.33). Only a tendency toward an inverse correlation with weight (r=-0.27) or body mass index (r=-0.26) was observed. No significant correlation with the pubertal stage was found. When chronological and bone ages were divided into intervals, a significant reduction of the AP angle was observed only in patients older than 14 years compared with those younger than 10 years of age. In this study, we propose that the AP angle should be considered to be normal if it varies between 148 and 155. We conclude that the normal AP angle does not depend on sex; however, it tends to decrease with stature, and chronological and bone ages. In the normal weight range also, the AP angle decreases, contrasting with our previous findings in obese adolescents, in which the AP angle increases with the severity of obesity. PMID- 17909335 TI - Avascular necrosis in acute and acute-on-chronic slipped capital femoral epiphysis. AB - This study reviews the factors affecting the development of avascular necrosis (AVN) after slipped capital femoral epiphysis. Seventy-three patients (n=73) with ninety-two (n=92) hips were included in the study. Out of these 92, 16 were acute, 43 acute-on-chronic, 29 chronic, and four were preslips. Postoperative radiographs showed a reduction in 18 (19.5%) slips. The mean preoperative slip angle was 32 degrees (range 9-76) compared with the postoperative slip angle of 29.9 degrees (range 10-75 degrees ) (P=0.004). Four patients developed AVN. Our results showed that intraoperative reduction of the slip (P<0.001) was significantly related to the development of AVN and was also associated with poor functional outcome. PMID- 17909336 TI - What follow-up is required for children with a family history of developmental dysplasia of the hip? AB - To determine what follow-up is needed for children with a family history of developmental dysplasia of the hip (DDH) but who have had a normal clinical and ultrasound examination at 6 weeks of age. A retrospective review of medical and imaging records of all children with a family history of DDH referred to the hip screening clinic over a 5-year period. An absolute acetabular index value of greater than 30 degrees or gross asymmetry in acetabular index values was used as an indication of occult acetabular dysplasia. One hundred and eighty-one children were identified as having had a normal ultrasound scan with a positive family history of DDH. Two (1.1%) children had acetabular dysplasia on their radiograph performed at 9-12 months of age. The dysplasia, however, resolved in both cases with simple observation and both children have now been discharged from further follow-up. We do not feel that it is necessary for children with a positive family history of DDH to have a radiograph of their pelvis at 1 year of age if they have had a normal clinical and ultrasound examination at 6 weeks of age and such children can safely be discharged from further follow-up at that time. PMID- 17909337 TI - The effect of tibial diaphyseal lengthening on the longitudinal growth of the tibia. AB - Limb lengthening by tibial callotasis is usually performed in the metaphysis but may cause growth inhibition. Is diaphyseal lengthening more advantageous? Sixteen immature rabbits underwent 30% diaphyseal lengthening by tibial callotasis. The tibial length was measured on radiographs at the end of the distraction period and after an additional 5 weeks. The proximal and distal growth plates were assessed histomorphometrically. Osteotomy stimulated tibial elongation; however, combined with diaphyseal lengthening the stimulation was suppressed resulting in longitudinal growth that matched the control side. In longer lengthenings of limbs diaphyseal callotasis may be more advantageous than metaphyseal by not inhibiting longitudinal growth. PMID- 17909338 TI - A 5-year follow-up study after knee disarticulation in two cases of Gollop Wolfgang complex. AB - The Gollop-Wolfgang Complex is a very rare anomaly, which has its essential features in congenital absence of the tibia and ipsilateral bifurcation of the femur. Surgical treatment of two patients with a follow-up of 5 years is reported. PMID- 17909339 TI - Treatment of acute salmonella epiphyseal osteomyelitis using computed tomography guided drainage in a child without sickle cell disease. AB - Salmonella osteomyelitis occurs infrequently in children without sickle cell disease. Similarly, acute osteomyelitis of the epiphysis has been rarely reported. We present a case of primary epiphyseal osteomyelitis caused by Salmonella in the distal femur of an otherwise healthy 17-month-old child. Before isolating an organism, parenteral nafcillin provided ineffective clinical, radiographic, and laboratory responses. Repeated fluoroscopic-guided percutaneous surgical drainages allowed for identification of the Salmonella, but did not resolve the epiphyseal infection, as the infection focus was missed. In the effort to eradicate the infection yet minimize further trauma to the epiphysis, computed tomography-guided drainage was performed and the infection subsequently resolved. Owing to its greater localization accuracy and minimal invasiveness, the computed tomography-guided intervention allowed for precise drainage without compromising the contiguous growth plate. At latest follow-up, the patient was ambulating well, had a normal knee examination, and had no evidence of leg length discrepancy or growth disturbance. PMID- 17909340 TI - Pseudomonas stutzeri knee arthritis in a child: case report and review. AB - We report a child with Pseudomonas stutzeri-associated right knee arthritis following knife puncture wound. Only four children with P. stutzeri-associated infections have been reported in the English literature in the last 40 years of whom one suffered from calcaneal osteomyelitis caused by this pathogen. In both cases, the suggested mechanism of the infection was local rather than bacterium invasion. PMID- 17909341 TI - Severe bilateral rachitic genu valgum in patients with nonbullous congenital ichthyosiform erythroderma: a report of two cases and review of literature. AB - We present two cases of ichthyosiform erythroderma associated with severe bilateral genu valgum. Other musculoskeletal features associated with this condition are described. The details and outcome of operative intervention for the correction of the deformities are discussed. The disturbances of the metabolism of vitamin D and medical management are discussed. A review of literature is presented. PMID- 17909342 TI - Aneurysmal bone cyst of the paediatric shoulder girdle: a case series and literature review. AB - Limited research has analysed paediatric shoulder girdle aneurysmal bone cyst management and outcomes. This study analysed locations affected, investigations, treatments and recurrence in children treated at the London Bone Tumour Unit between 1998 and 2004 and in English and French literature between 1956 and 2004. The proximal humerus and clavicle are most frequently affected whereas scapula involvement is rare. Radiographs, computed tomography and MRI are valuable. Sole curettage of clavicle and scapula lesions has low recurrence rates. Proximal humerus lesions recur most frequently. Curettage alone or with cementation are the most appealing treatments but are associated with significant recurrence. PMID- 17909343 TI - Atlantoaxial rotatory fixation owing to neck burn. AB - The purpose of this paper is to present a case of severe atlantoaxial rotatory fixation owing to a previously unreported etiology, and to discuss its pathogenesis and management. Conservative measures were unable to prevent progression, thus requiring surgical intervention. PMID- 17909344 TI - Cervical spine stenosis in chondrodysplasia punctata. AB - Chondrodysplasia punctata (CDP) is a rare skeletal dysplasia characterized by stippled epiphyses during infancy. The frequency is probably underdiagnosed because of the large heterogeneity in this group. Many genotypic variations exist. Although cervical instability is commonly seen in many skeletal dysplasias, cervical spine stenosis associated with CDP is very rare. We report a boy with phenotypic features of brachytelephalangic chondrodysplasia punctata (BCDP) who had severe cervical spine stenosis successfully corrected by vertebrectomies of C6 and C7 with a fibular strut graft. We discuss the significance of this association. PMID- 17909345 TI - Audit of elective paediatric clinic referrals from primary care February 2002 September 2006. AB - Our large district general hospital organized additional Waiting List Initiative clinics for paediatric orthopaedic referrals. A prospective audit examined pathology, investigations and disposal, and the implications of Waiting List Initiative clinics, particularly the increased workload for other specialties and disciplines. Variations of normal development were the most common presenting complaints, with the majority discharged following initial consultation. Local protocols, providing guidance to primary care physicians, covered common paediatric orthopaedic problems. We compared these guidelines with the referrals received to establish the referral quality and quantify the extra work generated. Few examples of inappropriate referrals were found, suggesting that referring physicians utilize the guidelines. PMID- 17909346 TI - The orthopedic and renal manifestations of idiopathic carpal tarsal osteolysis. AB - Idiopathic carpal tarsal osteolysis (ICTO) is a rare congenital disorder that results in the destruction and resorption of bone, leading to severe functional deficits and cosmetic deformities. This report includes a literature review describing the orthopedic and renal manifestations of ICTO. An additional case report of ICTO with atypical features is included. PMID- 17909347 TI - Mesothelioma: benefit from surgical resection is questionable. PMID- 17909348 TI - Role of carbon ion therapy for stage I NSCLC using a regimen of four fractions over week. PMID- 17909349 TI - Role of the wnt signaling pathway and lung cancer. PMID- 17909351 TI - ERCC1 mRNA expression is not associated with response and survival after platinum based chemotherapy regimens in advanced non-small cell lung cancer. AB - BACKGROUND: Platinum-based therapy is pivotal to the treatment of advanced non small cell lung Cancer (NSCLC). Excision repair cross-complementation group 1 (ERCC1) is a key component of the platinum-DNA repair machinery responsible for nucleotide excision repair. We sought to determine the influence of ERCC1 mRNA expression in advanced NSCLC on chemotherapy response, toxicity, and survival after platinum-based chemotherapy. METHODS: Patients randomized to a phase III trial of platinum-based chemotherapy were eligible for inclusion. Formalin-fixed paraffin-embedded tumor biopsies were retrieved for mRNA extraction and purification before quantitative real-time polymerase chain reaction analysis using Taqman technology. Expression data were correlated with treatment response, toxicity, and overall survival. RESULTS: Sixty-six patients were enrolled. No statistically significant relationship existed between ERCC1 mRNA expression and response to chemotherapy (p = 0.794) or hematological toxicity. No statistically significant difference in median survival was demonstrated according to ERCC1 expression (high expression, 415 days, 95% confidence interval [95%CI]: 197-633 days; low expression, 327 days [95%CI: 211-433 days]; p = 0.801). High ERCC1 mRNA expression was associated with a hazard ratio for death of 0.96 (95% CI 0.919 1.004; p = 0.08). CONCLUSION: In contrast to recent publications, ERCC1 mRNA expression in our study did not favor a prognostically better outcome after platinum-based chemotherapy in advanced NSCLC. We explore potential reasons for this, including the need for cautious interpretation of mRNA expression data from archival materials and highlight the need for additional translational research linking gene expression with a promising ERCC1 polymorphism. PMID- 17909352 TI - Postoperative prognosis in patients with non-small cell lung cancer according to the method of initial detection. AB - INTRODUCTION: In this study, we investigated the difference in the surgical results of non-small cell lung cancer according to the method of initial detection. METHODS: We reviewed the medical records of 796 patients who underwent pulmonary resection for non-small cell lung cancer between 1994 and 2005. The subjects consisted of 171 patients whose cancer was detected by a medical checkup or mass health screening (group I), 316 patients who were under evaluation for other diseases or with symptoms related to other diseases (group II), and 309 patients with lung cancer-related symptoms (group III). The mean ages of the three groups were 63.2, 69.7, and 68.2 years old, respectively, with group I being significantly younger than the other groups. The proportion of women in the symptomatic group was significantly lower than that of men. RESULTS: Pathologic stage I lung cancer was found in 112 (65.5%), 209 (65.2%), and 110 (35.6%) patients in groups I, II, and III, respectively. In comparison with stage II-IV cancer, stage I cancer was diagnosed more frequently in group I. According to the histologic type, adenocarcinoma was found in 132 patients (77.2%) in group I. However, squamous cell carcinoma was detected in only 27 patients (15.8%) in group I. The overall 5-year survival rates were 71.9%, 60.2%, and 48.0% in groups I, II, and III, respectively. Groups I and II had significantly better prognoses than group III. CONCLUSION: Groups I and II had favorable prognoses, and the presence of symptoms related to lung cancer was a significantly unfavorable prognostic factor independent of all other factors. PMID- 17909350 TI - Diagnostic accuracy of MALDI mass spectrometric analysis of unfractionated serum in lung cancer. AB - PURPOSE: There is a critical need for improvements in the noninvasive diagnosis of lung cancer. We hypothesized that matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) analysis of the most abundant peptides in the serum may distinguish lung cancer cases from matched controls. PATIENTS AND METHODS: We used MALDI MS to analyze unfractionated serum from a total of 288 cases and matched controls split into training (n = 182) and test sets (n = 106). We used a training-testing paradigm with application of the model profile defined in a training set to a blinded test cohort. RESULTS: Reproducibility and lack of analytical bias was confirmed in quality-control studies. A serum proteomic signature of seven features in the training set reached an overall accuracy of 78%, a sensitivity of 67.4%, and a specificity of 88.9%. In the blinded test set, this signature reached an overall accuracy of 72.6 %, a sensitivity of 58%, and a specificity of 85.7%. The serum signature was associated with the diagnosis of lung cancer independently of gender, smoking status, smoking pack-years, and C reactive protein levels. From this signature, we identified three discriminatory features as members of a cluster of truncated forms of serum amyloid A. CONCLUSIONS: We found a serum proteomic profile that discriminates lung cancer from matched controls. Proteomic analysis of unfractionated serum may have a role in the noninvasive diagnosis of lung cancer and will require methodological refinements and prospective validation to achieve clinical utility. PMID- 17909353 TI - Restaging of mediastinal nodes with transbronchial needle aspiration after induction chemoradiation for locally advanced non-small cell lung cancer. AB - INTRODUCTION: Selecting the appropriate treatment strategy for patients with locally advanced non-small cell carcinoma (NSCLC) is of utmost importance to determine patient outcome. Previous studies have shown that nodal down-staging after induction therapy and definitive local irradiation in these patients better predict survival when combined with surgery. However, nodal restaging can be technically difficult. We investigated the role of transbronchial needle aspiration (TBNA) in mediastinal restaging of patients who had completed induction cytotoxic therapy. METHODS: A total of 14 patients with proven stage IIIa-N2 NSCLC who received chemotherapy or chemo-radiotherapy as induction regimen between 2005 and 2006 were studied. Outpatient flexible bronchoscopy with TBNA was performed in all patients under local anesthesia, and 17 TBNA procedures were performed. TBNA results were matched against the histopathology of surgical specimens. RESULTS: Seventeen lymph nodes in 14 patients who had undergone induction therapy were sampled. Positron emission tomography (PET) scan results of 11 patients were also available for comparison. All positive TBNA procedures had positive PET scans. However, for five patients with lymph nodes measuring 9 to 17 mm, the PET scans were falsely positive, as mediastinoscopy and subsequent surgically resected lymph nodes revealed no tumor. TBNA achieved a correct diagnosis in 71% of patients who underwent mediastinal restaging and obviated further need for invasive procedures in 35%. CONCLUSION: For patients presenting with locally advanced NSCLC who are surgical candidates after induction chemo- and/or radiotherapy, TBNA should be considered as the initial procedure of choice for restaging of the mediastinum. PMID- 17909354 TI - Carbon ion radiotherapy for stage I non-small cell lung cancer using a regimen of four fractions during 1 week. AB - BACKGROUND: A phase I/II study was first conducted for the treatment of stage I non-small cell lung cancer (NSCLC) from 1994 to 1999 to determine the optimal dose. On the basis on the results, a phase II study using a regimen of four fractions during 1 week was performed. The purpose of the present study was to determine the local control and 5-year survival rates. METHODS: From December 2000 to November 2003, 79 patients with 80 primary lesions were treated. Using a fixed dose of 52.8 GyE for stage IA NSCLC and 60.0 GyE for stage IB NSCLC in four fractions during 1 week, the primary tumors were irradiated with carbon beams alone. The average age of the patients was 74.8 years. Sixty-two (78.5%) of these patients were medically inoperable. Local control and survival were determined using the Kaplan-Meier method. The data were statistically processed using the log-rank test. RESULTS: All patients were observed for a minimum of 3 years or until death, with a median follow-up time of 38.6 months, ranging from 2.5 to 72.2 months. The local control rate for all patients was 90% (T1: 98%, T2: 80%). The patients' 5-year lung cancer-specific survival rate was 68% (IA: 87%, IB: 42%). The overall survival was 45% (IA: 62%, IB: 25%). Half of the deaths were attributable to intercurrent diseases. No toxic reactions in the lung greater than grade 3 were detected. CONCLUSION: Carbon ion beam radiotherapy with a regimen of four fractions during 1 week has been proven as a valid alternative to surgery for stage I NSCLC and to offer particular benefits, especially for elderly and inoperable patients. PMID- 17909355 TI - Phase II multicenter trial with carboplatin and gemcitabine induction chemotherapy followed by radiotherapy concomitantly with low-dose paclitaxel and gemcitabine for stage IIIA and IIIB non-small cell lung cancer. AB - INTRODUCTION: The optimal combination of concomitant radiotherapy (RT) and chemotherapy in stage III unresectable non-small cell lung cancer (NSCLC) remains unclear. The role of induction chemotherapy with carboplatin/gemcitabine regimen has not been established in stage III NSCLC. METHODS: Forty-two stage III NSCLC patients, 41 assessable, with a median age of 60 years and good performance status, entered this trial between January 2003 and November 2004. They received carboplatin area under the curve 5 on day 1 and gemcitabine 1000 mg/m2 on days 1 + 8 every 3 weeks for two cycles, followed on day 50 by RT 60 Gy, concomitantly with paclitaxel 50 mg/m2 and gemcitabine 100 mg/m2 on days 1 + 8 every 3 weeks for two cycles. RESULTS: After induction, the partial response (PR) was 73.1% and stable disease was 24.4%. Disease progressed in one patient. After RT and paclitaxel/gemcitabine, 22% achieved a complete response and 73% a PR, and 5% had disease progression. The median survival was 25 months, the 1-year survival rate was 73.2%, and the 2-year survival rate was 50.5%. During concomitant RT and chemotherapy, grade 3 neutropenia, thrombocytopenia, and anemia occurred in eight, three, and three patients, respectively, and grade 4 neutropenia and thrombocytopenia in one patient each. One patient developed an esophageal fistula and died shortly after, which was considered a grade 5 toxicity; one patient developed grade 4 interstitial pneumonitis, and three patients developed grade 3 esophagitis. CONCLUSION: This regimen appears to be effective and was well tolerated. Further studies using this approach are warranted in patients with stage III NSCLC. PMID- 17909356 TI - Docetaxel and exisulind in previously treated non-small cell lung cancer (NSCLC) patients: a multicenter, phase II clinical trial. AB - PURPOSE: This multicenter, phase II clinical trial was conducted to evaluate the activity of the combination of docetaxel and exisulind in advanced non-small cell lung cancer (NSCLC) patients who failed a prior platinum-containing regimen. PATIENTS AND METHODS: Patients with measurable disease and adequate organ function received exisulind (250 mg) given orally, twice daily, and docetaxel (36 mg/m) administered intravenously on days 1, 8, and 15 of a 4-week cycle for up to six cycles. In the absence of disease progression or intolerable side effects, patients continued taking 250 mg of exisulind orally, twice daily. RESULTS: Thirty-three patients (median age 60 years; range 34-77; median performance status 1) were enrolled. There were no objective responses documented. Sixteen patients [48%, 95% confidence interval (CI): 31%-66%] had stable disease after 8 weeks of treatment. Median progression-free survival (PFS) was 2.1 months (95% CI: 1.5-3.2 months); median overall survival time was 8.0 months (range 0.2-25.9 months). Toxicity was moderate, with dose adjustment for adverse event/toxicity required for docetaxel or exisulind in 13 (39.3%) patients. Grade 3/4 lymphopenia, neutropenia, and anemia occurred in 48.5%, 12.1%, and 9.1% of patients, respectively. Grade 3 or greater toxicity was seen in 12.1%, 6.1%, and 3% of patients for nausea/vomiting, dyspnea, and abdominal pain, respectively. CONCLUSIONS: Treatment with exisulind and weekly docetaxel was not active in NSCLC patients who failed a prior platinum-containing regimen. Further study of this combination does not seem warranted. PMID- 17909358 TI - Targeting Lewis Y (Le(y)) in small cell lung cancer with a humanized monoclonal antibody, hu3S193: a pilot trial testing two dose levels. AB - INTRODUCTION: Lewis Y (Le(y)) is a blood group antigen with robust expression on the surface of epithelial tumors, including small cell lung cancer (SCLC), making it a potential target for antibody-based immunotherapy. 3S193, an immunoglobulin G3 monoclonal antibody, has demonstrated superior specificity, affinity, and cytotoxicity over other anti-Le(y) antibodies. A phase I trial of humanized 3S193 (hu3S193) with dosing up to 40 mg/m2 demonstrated tumor targeting without serious toxicities or the development of human anti-human antibodies. METHODS: We tested the targeting and pharmacokinetics of hu3S193 in patients with SCLC. Eligibility required progressive SCLC treated with up to three previous chemotherapy regimens, measurable disease not previously irradiated, and tumor samples positive for 3S193 by immunohistochemistry. Patients received four weekly injections of hu3S193, five patients at 10 mg/m2 and five patients at 20 mg/m2. The first and fourth injections were radiolabeled with indium-111 for gamma camera imaging. RESULTS: Of 40 patients screened, 25 of 34 (74%) assessable SCLC tumor samples were 3S193 positive by immunohistochemistry. Ten patients were treated with hu3S193; nine completed all four injections. All fluorodeoxyglucose (FDG)-avid lesions >2 cm were visualized on antibody single-photon emission computed tomography. Some lesions overlying vascular structures could not be visualized. No difference was noted in imaging or pharmacokinetics between the first and fourth injections. Toxicities included grade 2 urticaria (n = 1), grade 1 vomiting (n = 2), and grade 2 hypertension (n = 1) transiently after infusion at the higher dose. CONCLUSIONS: Given the strong tumor targeting, particularly at the higher dose, the favorable toxicity profile, and the potential for immunomodulatory effects, hu3S193 warrants further investigation in SCLC. PMID- 17909357 TI - Comparison of docetaxel- and vinca alkaloid-based chemotherapy in the first-line treatment of advanced non-small cell lung cancer: a meta-analysis of seven randomized clinical trials. AB - INTRODUCTION: To compare the impact on overall survival (OS) of docetaxel-based chemotherapy versus vinca alkaloid-based regimens for first-line therapy of advanced non-small cell lung cancer. METHODS: A meta-analysis of all randomized, controlled trials comparing docetaxel- and vinca alkaloid-based chemotherapy was undertaken using MEDLINE, CANCERLIT, MEDSCAPE, Google Scholar, the Cochrane Library, the National Institutes of Health randomized, controlled trials register, and conference proceedings, supplemented by information from clinical study reports. All published and unpublished randomized, controlled trials (in any language) were included. Analysis was based on pooling individual logarithms of the hazard ratio for OS and the odds ratio (OR) for safety. RESULTS: From eight potentially eligible trials, seven were selected (n = 2867). Docetaxel was administered with a platinum agent (three trials), with gemcitabine (two trials), or as monotherapy (two trials). Vinca alkaloid (vinorelbine [six trials] and vindesine [one trial]) was administered with cisplatin (six trials) or alone (one trial). The pooled estimate for OS showed an 11% improvement in favor of docetaxel (hazard ratio = 0.89; 95% confidence interval: 0.82-0.96; p = 0.004). Sensitivity analyses considering only vinorelbine as a comparator or only the doublet regimens showed similar improvements. Grade 3/4 neutropenia and grade 3/4 serious adverse events were less frequent with docetaxel- versus vinca alkaloid based regimens (OR = 0.59; 95% confidence interval: 0.38-0.89; p = 0.013 and OR = 0.68; 95% confidence interval: 0.55-0.84; p < 0.001, respectively). CONCLUSION: According to this meta-analysis, docetaxel is superior to vinca alkaloid-based regimens in terms of OS and safety for first-line therapy of advanced non-small cell lung cancer. PMID- 17909359 TI - Irinotecan-cisplatin therapy for patients with extensive-stage small cell lung cancer: use patterns among American medical oncologists 2000-2006. AB - BACKGROUND: Etoposide-platinum doublet therapy has been the American standard first-line management for patients with small cell lung cancer for more than a decade. In the 1990s, Japanese investigators developed a regimen of irinotecan cisplatin for extensive-stage patients and, in one head-to-head trial, the irinotecan therapy produced a statistically significant survival advantage compared with etoposide and cisplatin. We were interested in how American oncologists integrate clinical trial data into their current prescribing practices and studied case-based prescribing recommendations as a surrogate for current clinical practice. We evaluated Network for Medical Communications and Research data from 7 years of live physician meetings to assess changes in use patterns for irinotecan cisplatin since the initial report of the Japanese clinical trial. METHODS: Data from 38 meetings involving 2174 medical professional attendees (MD; DO; MD PhD) were reviewed. Attendees were given a case scenario and asked to select the treatment option that they felt was most appropriate for the hypothetical setting provided. At each meeting, responses were collected electronically and immediately displayed to the attendees. Aggregate response data are held in the Network for Medical Communications and Research database. RESULTS: During the 7-year study period, regimens including etoposide and platinum have consistently been the most frequently recommended for small cell lung cancer therapy by the meeting participants. The selection of irinotecan-cisplatin by American oncologists was initially infrequent, with a modest, transient impact of a plenary session presentation of the Japanese phase III trial data at the 2000 meeting of the American Society of Clinical Oncology. Publication of the phase III trial findings in the New England Journal of Medicine in early 2002 stimulated a marked increase in irinotecan-cisplatin prescribing with a commensurate decrease in the selection of etoposide and cisplatin therapy. Since 2002, the prescribing patterns have shown an accelerating return to the use of etoposide and platinum despite continuing study of irinotecan-platinum regimens. CONCLUSIONS: Statistically significant advantages demonstrated in a single phase III clinical trial may have only a modest and transient impact on American oncologists' prescribing behaviors. Factors other than the phase III trial results themselves play a role in the likelihood that prescribing behavior will evolve over time. Corroborating trial data among unambiguously relevant populations will be necessary to stimulate a change in standard treatment paradigms. PMID- 17909360 TI - Prognostic factors in the treatment of malignant pleural mesothelioma at a large tertiary referral center. AB - INTRODUCTION: Most studies describing the natural history and prognostic factors for malignant pleural mesothelioma antedate accurate pathologic diagnosis, staging by computed tomography, and a universal staging system. We conducted a large single-institution analysis to identify prognostic factors and assess the association of resection with outcome in a contemporary patient population. METHODS: Patients with biopsy-proven malignant pleural mesothelioma at our institution were identified and clinical data were obtained from an institutional database. Survival and prognostic factors were analyzed by the Kaplan-Meier method, log-rank test, and Cox proportional hazards analysis. A p value <0.05 was considered statistically significant. RESULTS: From 1990 to 2005, 945 patients were identified: 755 men, 190 women; median age, 66 years (range, 26-93). Extrapleural pneumonectomy was performed in 208 (22%), pleurectomy/decortication in 176 (19%). Operative mortality was 4% (16/384). Multimodality therapy including surgery was associated with a median survival of 20.1 months. Significant predictors of overall survival included histology, gender, smoking, asbestos exposure, laterality, surgical resection by extrapleural pneumonectomy or pleurectomy/decortication, American Joint Committee on Cancer stage, and symptoms. A Cox model demonstrated a hazard ratio of 1.4 without surgical resection when controlling for histology, stage, gender, asbestos exposure, smoking history, symptoms, and laterality (p = 0.003). CONCLUSIONS: In addition to tumor histology and pathologic stage, predictors of survival include gender, asbestos exposure, smoking, symptoms, laterality, and clinical stage. Surgical resection in a multimodality setting was associated with improved survival. PMID- 17909361 TI - Rapid growth of malignant pleural mesothelioma. PMID- 17909362 TI - Occurrence of pleural masses in a chronic pleural pyothorax. PMID- 17909364 TI - Plasmacytoma of bronchus treated by radical radiotherapy--a rare case with four and a half years follow up. AB - Plasmacytoma of the bronchus is a very rare plasma cell neoplasm affecting the bronchus. Here we report a case of plasmacytoma of the bronchus treated by radical radiotherapy in July 2002. The tumor responded very well to treatment and showed a slow but sustained regression in the size over two years. Presently, he has completed four and a half years of follow-up and is free of disease. PMID- 17909363 TI - Endobronchial ultrasonography: current status and future directions. AB - Endobronchial ultrasonography (EBUS) has emerged as a new diagnostic tool that allows the bronchoscopist to see beyond the airway. The radial probe EBUS was first introduced to evaluate the airway structure, which has been shown to be useful for identifying the extent of tumor invasion in the central airway. With advance in technology, smaller radial probes are now available that are capable of visualizing peripheral lung nodules. EBUS is also used as a tool to assist in a biopsy in respiratory diseases. The radial probe EBUS-guided transbronchial needle aspiration (TBNA) increases the yield of TBNA of mediastinal processes. By the use of the ultra-miniature probe EBUS along with the guide sheath, peripheral lung lesions can be accessed without the exposure to radiation. However, it is still not a real-time procedure with target visualization. The newest development is the convex probe EBUS (CP-EBUS) with a curvilinear electronic transducer on the tip of a flexible bronchovideoscope. CP-EBUS allows real-time EBUS-guided TBNA. Although the main indication for EBUS-TBNA is lymph node staging, it can also be used for diagnosis of intrapulmonary tumors, of unknown hilar and/or mediastinal lymphadenopathy, and of mediastinal tumors. To date, there are no reports of complications related to EBUS-guided TBNA. It is a novel approach that has a good diagnostic yield with excellent potential in assisting safe and accurate diagnostic interventional bronchoscopy. The aim of this review is to highlight the current status of the different EBUS techniques available and to discuss the future direction of EBUS. PMID- 17909367 TI - Using inverse probability-weighted estimators in comparative effectiveness analyses with observational databases. AB - Inverse probability-weighted estimation is a powerful tool for use with observational data. In this article, we describe how this propensity score-based method can be used to compare the effectiveness of 2 or more treatments. First, we discuss the inherent problems in using observational data to assess comparative effectiveness. Next, we provide a conceptual explanation of inverse probability-weighted estimation and point readers to sources that address the method in more formal, technical terms. Finally, we offer detailed guidance about how to implement the estimators in comparative effectiveness analyses. PMID- 17909365 TI - Malignant mediastinal tumor with bone formation--mesothelioma or sarcoma? AB - Mesothelioma can occur in different variants, some of which are difficult or impossible to differentiate from sarcomas. There are scattered reports of sarcomatous mesotheliomas that have osteogenic properties. Here, we report a 57 year old man who presented with a mediastinal tumor containing scattered irregular calcifications with some scattered pleural thickening of the right pleura. Biopsy showed a sarcoma with bone formation. The man was born in the Turkish village of Karain, where the incidence of mesothelioma is extremely high, and a sarcomatous mesothelioma was therefore diagnosed. Since the tumor was pressing against the large vessels and heart, a debulking was performed, followed by Pemetrexed and Carboplatin treatment. However, the tumor grew rapidly and spread to the pleura, involved the heart, and the patient succumbed. This is to our knowledge the first report of a sarcomatous mesothelioma with bone formation from environmental exposure to mineral fibers. PMID- 17909368 TI - A simulation-based evaluation of methods to estimate the impact of an adverse event on hospital length of stay. AB - INTRODUCTION: We used agent-based simulation to examine the problem of time varying confounding when estimating the effect of an adverse event on hospital length of stay. Conventional analytic methods were compared with inverse probability weighting (IPW). METHODS: A cohort of hospitalized patients, at risk for experiencing an adverse event, was simulated. Synthetic individuals were assigned a severity of illness score on admission. The score varied during hospitalization according to an autoregressive equation. A linear relationship between severity of illness and the logarithm of the discharge rate was assumed. Depending on the model conditions, adverse event status was influenced by prior severity of illness and, in turn, influenced subsequent severity. Conditions were varied to represent different levels of confounding and categories of effect. The simulation output was analyzed by Cox proportional hazards regression and by a weighted regression analysis, using the method of IPW. The magnitude of bias was calculated for each method of analysis. RESULTS: Estimates of the population causal hazard ratio based on IPW were consistently unbiased across a range of conditions. In contrast, hazard ratio estimates generated by Cox proportional hazards regression demonstrated substantial bias when severity of illness was both a time-varying confounder and intermediate variable. The direction and magnitude of bias depended on how severity of illness was incorporated into the Cox regression model. CONCLUSIONS: In this simulation study, IPW exhibited less bias than conventional regression methods when used to analyze the impact of adverse event status on hospital length of stay. PMID- 17909369 TI - Evaluating the validity of an instrumental variable study of neuroleptics: can between-physician differences in prescribing patterns be used to estimate treatment effects? AB - BACKGROUND: Postmarketing studies of prescription drugs are challenging because prognostic variables that determine treatment choices are often unmeasured. In this setting, instrumental variable (IV) methods that exploit differences in prescribing patterns between physicians may be used to estimate treatment effects; however, IV methods require strong assumptions to yield consistent estimates. We sought to explore the validity of physician-level IV in a comparative study of short-term mortality risk among elderly users of conventional versus atypical antipsychotic medications (APM). METHODS: We studied a cohort of patients initiating APMs in Pennsylvania who were eligible for Medicare and a state-funded pharmaceutical benefit plan. The IV was defined as the type of the APM prescription written by each physician before the index prescription. To evaluate whether the IV was related to other therapeutic decisions that could affect mortality, we explored the association between the instrument and 2 types of potentially hazardous coprescriptions: a tricyclic antidepressant (TCA) not recommended for use in the elderly or a long-acting benzodiazepine. To insure that the IV analysis was not biased by case-mix differences between physicians, we examined the associations between the observed patient characteristics and the IV. RESULTS: The cohort consisted of 15,389 new users of APMs. Our multivariable model indicated that physicians who had most recently prescribed a conventional APM were not significantly more or less likely to coprescribe a potentially hazardous TCA [odds ratio (OR), 0.78; 95% confidence interval (CI), 0.58-1.02] but were less likely to prescribe a long-acting benzodiazepine (OR, 0.57; 95% CI, 0.45-0.72) with their current APM prescription. The association between long-acting benzodiazepine prescribing and APM preference was no longer significant when the analysis was restricted to primary care physicians (OR, 0.84; 95% CI, 0.62-1.15). Multivariable regression indicated that important medical comorbidities (eg, cancer, hypertension, stroke) were unrelated to the IV. CONCLUSIONS: The previous APM prescription written by the physician was unassociated with major medical comorbidities in the current patient, suggesting that the IV estimates were not biased by case-mix differences between physicians. However, we did find that the IV was associated with the use of long acting benzodiazepines. This association disappeared when the study was restricted to the patients treated by primary care physicians. Our study illustrates how internal validation approaches may be used to improve the design of quasi-experimental studies. PMID- 17909370 TI - Heterogeneity and the interpretation of treatment effect estimates from risk adjustment and instrumental variable methods. AB - OBJECTIVES: To contrast the interpretations of treatment effect estimates using risk adjustment and instrumental variable (IV) estimation methods using observational data when the effects of treatment are heterogeneous across patients. We demonstrate these contrasts by examining the effect of breast conserving surgery plus irradiation (BCSI) relative to mastectomy on early stage breast cancer (ESBC) survival. METHODS: We estimated discrete time survival models for 6185 ESBC patients in the 1989-1994 Iowa Cancer Registry via IV estimation using 2 distinct instruments (distance of the patient's residence from the nearest radiation center, and local area BCSI rate) and controlling for cancer stage, grade, and location; age; comorbidity; hospital access; payer; diagnosis year; and area poverty level. We then estimated comparable risk adjustment survival models using linear probability methods with robust standard errors. RESULTS: Risk adjustment models yielded average survival estimates similar to trial results. With favorable BCSI selection, these estimates represent an upper bound of the true effect for patients receiving BCSI. IV estimates showed a BCSI survival risk for patients whose surgery choices were affected by the instruments and these estimates varied with the instrument specification. CONCLUSIONS: When treatment benefits are heterogeneous across patients, treatment effect estimates from observational data can still be useful to policymakers, but they must be interpreted correctly. Risk adjustment methods yield estimates that can assess whether the patients who received treatment benefited from the treatment, but the direction of bias must be considered. In contrast, IV estimates can assess the effect of treatment rate changes, but characteristics of patients whose choices were affected by the instruments must be considered when making such inferences. PMID- 17909371 TI - Methodologic challenges to studying patient safety and comparative effectiveness. AB - Studies of patient safety and comparative effectiveness entail unique methodologic challenges. These studies may be susceptible to systematic error, including selection bias, exposure misclassification, and outcome misclassification. They may also be vulnerable to random error, or confounding by a variable such as another drug, a disease, or the drug indication itself. Finally, special logistical issues can arise, including data access problems, difficulties in conveying the need for studies of certain interventions, and obstacles to gaining institutional review board approval. This article provides a conceptual overview of these methodologic issues. PMID- 17909373 TI - Use of propensity score technique to account for exposure-related covariates: an example and lesson. AB - BACKGROUND: In observational research, propensity score techniques can be used to account for baseline differences between compared therapies. Although propensity scores are used increasingly often, their limitations in settings without complete data may not be recognized. OBJECTIVES: We sought to evaluate the ability of propensity score matching to mitigate confounding by indication in an observational study of the effect of statin therapy on acute myocardial infarction (AMI). Matching was performed at random, and with propensity scores that incorporated a reduced or expanded set of variables. RESEARCH DESIGN/SUBJECTS: This was a propensity score matched cohort study using members of a health insurer database. MEASURES: Exposure to statin therapy was assessed at the beginning of follow-up with all cohort members being statin initiators or noninitiators, and the outcome of AMI was identified on the basis of claims codes. RESULTS: Matching on the basis of the propensity score provided results that are similar in magnitude to randomized clinical trials, suggesting that confounding was mitigated. However, matching on a propensity score created on a reduced set of variables yielded a result that suggested no effect of statin therapy, and demonstrated substantial imbalance on some variables that were not part of the propensity score. CONCLUSIONS: Propensity score matching can balance with respect to variables not explicitly included in the score, but external data are required to evaluate this. PMID- 17909372 TI - Increasing levels of restriction in pharmacoepidemiologic database studies of elderly and comparison with randomized trial results. AB - BACKGROUND: The goal of restricting study populations is to make patients more homogeneous regarding potential confounding factors and treatment effects and thereby achieve less biased effect estimates. OBJECTIVES: This article describes increasing levels of restrictions for use in pharmacoepidemiology and examines to what extent they change rate ratio estimates and reduce bias in a study of statin treatment and 1-year mortality. METHODS: : The study cohort was drawn from a population of seniors age 65 years and older enrolled in both Medicare and the Pennsylvania Pharmaceutical Assistance Contract for the Elderly (PACE) between 1995 and 2002. We identified all users of statins during the study period and assessed the time until death within 1 year. The following progressive restrictions were applied: (1) study incident drug users only, (2) choose a comparison group most similar to the intervention group, (3) exclude patients with contraindications, (4) exclude patients with low adherence, and (5) restrict to specific high-risk/low-risk subgroups represented in randomized trails (RCTs). RESULTS: The basic cohort comprised 122,406 statin users, who were on average 78 years old and predominantly white (93%) and showed an unadjusted rate ratio of 0.32 for statin users. When all 5 restrictions were applied (N = 11,673), the unadjusted rate ratio had increased to 0.72. Multivariable Cox regression adjusted rate ratios increased from 0.62 [95% confidence interval (CI), 0.58 0.66] to 0.79 (95% CI, 0.60-1.03). However, after the first 3 restrictions the effect size changed little. The final estimate is similar to that obtained as a pooled estimate of 3 pravastatin RCTs in patients age 65 years and older. We argue that restrictions 1 through 4 compromised generalizability little. CONCLUSIONS: In our example of a large database study, restricting to incident drug users, similar comparison groups, patients without contraindication, and to adherent patients was a practical strategy, which limited the effect of confounding, as these approaches yield results closer to those seen in RCTs. PMID- 17909374 TI - Using propensity scores subclassification to estimate effects of longitudinal treatments: an example using a new diabetes medication. AB - BACKGROUND: When using observational data to compare the effectiveness of medications, it is essential to account parsimoniously for patients' longitudinal characteristics that lead to changes in treatments over time. OBJECTIVES: We developed a method of estimating effects of longitudinal treatments that uses subclassification on a longitudinal propensity score to compare outcomes between a new drug (exenatide) and established drugs (insulin and oral medications) assuming knowledge of the variables influencing the treatment assignment. RESEARCH DESIGN/SUBJECTS: We assembled a retrospective cohort of patients with diabetes mellitus from among a population of employed persons and their dependents. METHODS: The data, from i3Innovus, includes claims for utilization of medications and inpatient and outpatient services. We estimated a model for the longitudinal propensity score process of receiving a medication of interest. We used our methods to estimate the effect of the new versus established drugs on total health care charges and hospitalization. RESULTS: We had data from 131,714 patients with diabetes filling prescriptions from June through December 2005. Within propensity score quintiles, the explanatory covariates were well-balanced. We estimated that the total health care charges per month that would have occurred if all patients had been continually on exenatide compared with if the same patients had been on insulin were minimally higher, with a mean monthly difference of $397 [95% confidence interval (CI), $218-$1054]. The odds of hospitalization were also comparable (relative odds, 1.02; 95% CI, 0.33-1.98). CONCLUSIONS: We used subclassification of a longitudinal propensity score for reducing the multidimensionality of observational data, including treatments changing over time. In our example, evaluating a new diabetes drug, there were no demonstrable differences in outcomes relative to existing therapies. PMID- 17909376 TI - Creating and synthesizing evidence with decision makers in mind: integrating evidence from clinical trials and other study designs. AB - BACKGROUND: Randomized controlled trials (RCTs) remain the accepted "gold standard" for determining the efficacy of new drugs or medical procedures. Randomized trials alone, however, cannot provide all the relevant information decision makers need to determine the relative risks and benefits when choosing the best treatment of individual patients or weighing the implications of particular policies affecting medical therapies. OBJECTIVES: To demonstrate the limitations of RCTs in providing the information needed by medical decision makers, and to show how information from observational studies can supplement evidence from RCTs. METHODS: Qualitative description of the limitations of RCTs in providing the information needed by medical decision makers, and demonstration of how evidence from additional sources can aid in decision making, using the examples of deciding whether a 60-year-old woman with mildly elevated blood pressure should take daily low-dose aspirin, and whether a hospital network should implement carotid artery surgery for asymptomatic patients. CONCLUSIONS: Even the most rigorously designed RCTs leave many questions central to medical decision making unanswered. Research using cohort and case-control designs, disease and intervention registries, and outcomes studies based on administrative data can all shed light on who is most likely to benefit from the treatment, and what the important tradeoffs are. This suggests the need to revise the traditional evidence hierarchy, whereby evidence progresses linearly from basic research to rigorous RCTs. This revised hierarchy recognizes that other research designs can provide important evidence to strengthen our understanding of how to apply research findings in practice. PMID- 17909375 TI - Adjustments for unmeasured confounders in pharmacoepidemiologic database studies using external information. AB - BACKGROUND: Nonexperimental studies of drug effects in large automated databases can provide timely assessment of real-life drug use, but are prone to confounding by variables that are not contained in these databases and thus cannot be controlled. OBJECTIVES: To describe how information on additional confounders from validation studies can help address the problem of unmeasured confounding in the main study. RESEARCH DESIGN: Review types of validation studies that allow adjustment for unmeasured confounding and illustrate these with an example. SUBJECTS: Main study: New Jersey residents age 65 years or older hospitalized between 1995 and 1997, who filled prescriptions within Medicaid or a pharmaceutical assistance program. Validation study: representative sample of Medicare beneficiaries. MEASURES: Association between nonsteroidal antiinflammatory drugs (NSAIDs) and mortality. RESULTS: Validation studies are categorized as internal (ie, additional information is collected on participants of the main study) or external. Availability of information on disease outcome will affect choice of analytic strategies. Using an external validation study without data on disease outcome to adjust for unmeasured confounding, propensity score calibration (PSC) leads to a plausible estimate of the association between NSAIDs and mortality in the elderly, if the biases caused by measured and unmeasured confounders go in the same direction. CONCLUSIONS: Estimates of drug effects can be adjusted for confounders that are not available in the main, but can be measured in a validation study. PSC uses validation data without information on disease outcome under a strong assumption. The collection and integration of validation data in pharmacoepidemiology should be encouraged. PMID- 17909377 TI - Comparison of meta-analytic results of indirect, direct, and combined comparisons of drugs for chronic insomnia in adults: a case study. AB - BACKGROUND: Our Center recently conducted a systematic review of the manifestations and management of chronic insomnia in adults. The efficacy and safety of benzodiazepines and nonbenzodiazepines, relative to placebo, were compared indirectly. OBJECTIVES: Determine how the results of indirect comparisons made in the review compare with the results of direct comparisons, as well as with estimates derived from Bayesian mixed treatment comparisons. Establish general appropriateness of the use of results of indirect or mixed treatment comparisons. METHODS: Treatments were compared using frequentist direct, indirect, and combined methods, as well as Bayesian direct and mixed methods. RESULTS: Estimates for comparisons tended to be clinically and statistically similar across methods. Estimates obtained through indirect comparisons were not biased and were similar to those obtained through direct analysis. CONCLUSIONS: Results of indirect comparisons made in the review, accurately reflected the current evidence. Frequentist and Bayesian methods of analysis of indirect comparisons should be considered when performing meta analyses. PMID- 17909378 TI - Improving depiction of benefits and harms: analyses of studies of well-known therapeutics and review of high-impact medical journals. AB - The issues of weighing benefits and harms and of shared decision-making have become increasingly important in recent years. There is limited knowledge and lack of adequate data on the most transparent method of communicating the information. In this article we discuss examples of communicating benefits and harms for well-known therapeutics, illustrating that relative risk estimates are not helpful for communicating the chance of experiencing adverse events. In addition, we show that asymmetric presentation of the data for benefits and harms is likely to bias toward showing greater benefits and diminishing the importance of the harms (or vice versa). We also present preliminary results of a brief review of high-impact medical journals that show limitations of current systematic reviews. In the review we found that every second published study does not discuss frequency data and 1 in 3 studies that report information on both benefits and harms does not report information in the same metric. We conclude that consistently depicting benefit and harm information in frequencies can substantially improve the communication of benefits and harms. Investigators should be requested to provide frequency data along with relative risk information in the publication of their scientific findings. Currently, even in the highest impact medical journals, evidence of benefits and harms is not consistently presented in ways that facilitate accurate interpretation. PMID- 17909379 TI - Cluster randomized trials: opportunities and barriers identified by leaders of eight health plans. AB - BACKGROUND: Cluster randomized trials (CRTs) offer unique advantages over standard randomized controlled clinical trials (RCTs) and observational methodologies, and may provide a cost-efficient alternative for answering questions about the best treatments for common conditions. OBJECTIVES: To describe health plan leaders' views on CRTs, identify barriers to conducting CRTs, and solicit recommendations for increasing the acceptability of CRTs. RESEARCH DESIGN: Qualitative in-depth telephone interviews with leaders from 8 health plans. SUBJECTS: : Thirty-four health plan leaders (medical directors, pharmacy directors, Institutional Review Board leaders, ethics leaders, compliance leaders, and others). MEASURES: Qualitative analysis of interview transcripts to identify barriers, factors influencing leaders' views, ethical issues, aspects of CRTs that appeal to leaders, and recommendations for increasing acceptability of CRTs. RESULTS: Multiple barriers were identified, including financial costs, concerns about stakeholders' perceptions of CRTs, impact on physicians' prescribing habits, and formulary changes. Most leaders recognized the potential value of studying the comparative effectiveness of therapeutics, and many stressed the need for head-to-head trials. Leaders' views would be influenced by variations in study design and implementation. Recommendations for increasing acceptability of CRTs included ensuring that the fiscal impact of a CRT be budget neutral, and that researchers educate stakeholders and decision-makers about CRTs. CONCLUSIONS: Overall, health plan leaders recognized the need for studies of the comparative effectiveness of therapeutics under real world conditions, and many expressed support for CRTs. However, researchers seeking to conduct CRTs in health plans are likely to face numerous barriers, and preparatory work will be essential. PMID- 17909380 TI - The comparative effectiveness and safety emerging methods symposium: a tribute to Harry A. Guess: father and consummate leader of pharmacoepidemiology. PMID- 17909381 TI - Design of cluster-randomized trials of quality improvement interventions aimed at medical care providers. AB - BACKGROUND: Randomized trials aimed at improving the quality of medical care often randomize the provider. Such trials are frequently embedded in health care systems with available automated records, which can be used to enhance the design of the trial. METHODS: We consider how available information from automated records can address each of the following concerns in the design of a trial: whether to randomize individual providers or practices; clustering of outcomes among patients in the same practice and its impact on study size; expected heterogeneity in adherence and the response to the intervention; eligibility criteria and the trade-offs between generalizability and internal validity; and blocking or matching to alleviate covariate imbalance across practices. RESULTS: Investigators can use available information from an automated database to estimate the amount of clustering of patients within providers and practices, and these estimates can inform the decision on whether to randomize at the level of the patient, the provider, or the practice. We illustrate calculation of the anticipated design effect for a proposed cluster-randomized trial and its implications for sample size. With available claims data, investigators can apply focused eligibility criteria to exclude subjects and providers with expected low compliance or lower likelihood of benefit, although possibly at some loss of generalizability. Chance imbalances in covariates are more likely when randomization occurs at the level of the practice than at the level of the patient, so we propose a matching score to limit such imbalances by design. CONCLUSIONS: Challenges to compliance, expected small effects, and covariate imbalances are particularly likely in cluster-randomized trials of quality improvement interventions. When such trials are embedded in medical systems with available automated records, use of these data can enhance the design of the trial. PMID- 17909382 TI - Designed delays versus rigorous pragmatic trials: lower carat gold standards can produce relevant drug evaluations. AB - BACKGROUND: Centralized administrative databases enable low-cost pragmatic randomized trials (PRTs) of drug effectiveness and safety. We simplified the PRT strategy by using designed delays (DD) to evaluate drug policies. OBJECTIVES: To reassess our DD trial of a cost-saving nebulizer-to-inhaler conversion policy and a proposed DD trial of reduced restrictions on Cox-2 inhibitors. RESEARCH DESIGN: We randomized 52 pairs of communities and clusters of physician practices to the policy either on time or after a 6-month delay. Our 2-stage qualitative reassessment comprised: (1) applying criteria for reporting PRTs and (2) assessing DD trials in 3 domains of responsibility: policymakers' decisions, researchers' decisions, and joint decisions involving negotiation. MEASURES: A draft checklist of 22 Consolidated Standards of Reporting Trials (CONSORT). Researchers' recollections of their degree of influence on decisions. RESULTS: DD trials deviated from ideal PRTs in the policymakers' domain: the policies affected mixtures of drugs, users, and illnesses, and implementation was not by strict protocol. Aspects negotiated by researchers and policymakers also deviated from ideal: length of delay; size and location of control group; unit of randomization; additional data collection; and communications to physicians. The DD trials complied better with CONSORT in the researchers' domain of analysis and interpretation. CONCLUSIONS: DD trials can be negotiated with policymakers. Low cost and simplicity of DD trials partly compensate for some limitations for evaluating drug safety and effectiveness. The ethics question of whether a DD is routine evaluation or research depends on its purpose and generalizability. PMID- 17909383 TI - Emerging methods in comparative effectiveness and safety: symposium overview and summary. AB - BACKGROUND: Interest in new methods for comparative effectiveness, drug and patient safety, and related studies is burgeoning. The advent of Medicare Part D for outpatient prescription drugs has drawn significant attention to the need for efficient ways to monitor the potential benefits and harms of pharmaceuticals. These trends prompted the Effective Health Care program at the Agency for Healthcare Research and Quality and its DEcIDE (Developing Evidence to Inform Decisions about Effectiveness) network to examine innovative approaches for such investigations through an invitational symposium in June 2006. RESULTS: Conference papers covered numerous points about ways to structure both interventional and database-oriented studies, particularly those concerned with adverse drug events, to avoid bias in those studies, and to apply advanced statistical tools to exploit the information from these studies to their fullest. Of particular importance are: (1) using new types of experimental designs, including cluster randomization, delayed designs, pragmatic trials, and practice based investigations that incorporate the natural variation of data from routine clinical practice; (2) finding efficient ways to use different types of databases eg, Department of Veterans Affairs files, Centers for Disease Control and Prevention surveillance files, Medicaid claims data, and state hospital data-for examining initiation, persistence, and adherence, and the benefits and adverse events of pharmaceutical use; and (3) inventing or refining ways to decrease the threats to validity of analyses relying on administrative or other observational data, particularly through propensity scoring, inverse probability weighting, risk adjustment, and direct or indirect methods for synthesizing comparative effectiveness information. PMID- 17909384 TI - Practice-based evidence study design for comparative effectiveness research. AB - OBJECTIVES: To describe a new, rigorous, comprehensive practice-based evidence for clinical practice improvement (PBE-CPI) study methodology, and compare its features, advantages, and disadvantages to those of randomized controlled trials and sophisticated statistical methods for comparative effectiveness research. RESEARCH DESIGN: PBE-CPI incorporates natural variation within data from routine clinical practice to determine what works, for whom, when, and at what cost. It uses the knowledge of front-line caregivers, who develop study questions and define variables as part of a transdisciplinary team. Its comprehensive measurement framework provides a basis for analyses of significant bivariate and multivariate associations between treatments and outcomes, controlling for patient differences, such as severity of illness. RESULTS: PBE-CPI studies can uncover better practices more quickly than randomized controlled trials or sophisticated statistical methods, while achieving many of the same advantages. We present examples of actionable findings from PBE-CPI studies in postacute care settings related to comparative effectiveness of medications, nutritional support approaches, incontinence products, physical therapy activities, and other services. CONCLUSIONS: Outcomes improved when practices associated with better outcomes in PBE-CPI analyses were adopted in practice. PMID- 17909385 TI - Studying prescription drug use and outcomes with medicaid claims data: strengths, limitations, and strategies. AB - Medicaid claims and eligibility data, particularly when linked to other sources of patient-level and contextual information, represent a powerful and under-used resource for health services research on the use and outcomes of prescription drugs. However, their effective use poses many methodological and inferential challenges. This article reviews strengths, limitations, challenges, and recommended strategies in using Medicaid data for research on the initiation, continuation, and outcomes of prescription drug therapies. Drawing from published research using Medicaid data by the investigators and other groups, we review several key validity and methodological issues. We discuss strategies for claims based identification of diagnostic subgroups and procedures, measuring and modeling initiation and persistence of regimens, analysis of treatment disparities, and examination of comorbidity patterns. Based on this review, we discuss "best practices" for appropriate data use and validity checking, approaches to statistical modeling of longitudinal patterns in the presence of typical challenges, and strategies for strengthening the power and potential of Medicaid datasets. Finally, we discuss policy implications, including the potential for the research use of Medicare Part D data and the need for further initiatives to systematically develop and optimally use research datasets that link Medicaid and other sources of clinical and outcome information. PMID- 17909386 TI - Assessment of adherence to and persistence on disease-modifying antirheumatic drugs (DMARDs) in patients with rheumatoid arthritis. AB - OBJECTIVE: Biologic disease-modifying antirheumatic drugs (DMARDs) are efficacious for treating rheumatoid arthritis (RA). However, measurements of relative effectiveness, including treatment adherence and persistence, are lacking. We evaluated adherence and persistence during new episodes of use of traditional and biologic DMARDs. METHODS: Using Tennessee Medicaid databases (1995-2004), we assembled a retrospective cohort of patients diagnosed with RA, and identified new episodes of use for 12 DMARD regimens. We evaluated persistence through survival analyses, and adherence within episodes through the medication possession ratio. A risk score was included in the analyses to account for measured confounders. RESULTS: We identified 14,932 patients with RA; 6018 patients had 10,547 episodes of new use of DMARDs. Considering methotrexate as the reference and after adjustment for measured confounders, episodes of new use of sulfasalazine [adjusted hazard ratio (aHR) = 1.59; 95% confidence interval (CI) = 1.47-1.72] and infliximab alone (aHR = 1.37, 95% CI = 1.09-1.73) were more likely to be discontinued; and new episodes of etanercept (aHR = 0.82, 95% CI = 0.73-0.92) and methotrexate + adalimumab (aHR = 0.63, 95% CI = 0.48-0.84) were less likely to be discontinued. Compared with methotrexate, adherence was higher for leflunomide, infliximab, etanercept, and adalimumab and lower for sulfasalazine and all combined therapies. CONCLUSIONS: We developed an approach to assess persistence on and adherence to the most common DMARD therapies. In this large cohort, persistence and adherence to leflunomide and most biologic DMARD therapies were at least comparable to methotrexate. Adherence was lower for sulfasalazine and all combined therapies. PMID- 17909387 TI - Out-of-pocket pharmacy expenditures for veterans under medicare part D. AB - OBJECTIVES: Because the VA pharmacy benefit is deemed equivalent coverage to Medicare Part D, veterans can use either or both of these Federal benefits. We sought to determine how these programs' different benefit structures and low income assistance thresholds would affect pharmacy out-of-pocket expenses for veterans. METHODS: We reviewed income and asset tests performed at the Salt Lake City VA in fiscal year 2005, and estimated the number of individuals, age 65 and older, who meet eligibility for Part D low-income assistance. Using past VA pharmacy utilization data, we estimated the difference in pharmacy out-of-pocket expenditures for veterans eligible for assistance through Medicare but not through the VA. RESULTS: The income and asset thresholds for low-income assistance through Part D were reached by 4127 veterans. From this group, we identified 926 veterans who had used the VA pharmacy during the prior year, who are ineligible for VA copayment waivers, and who qualify for premium waiver under Part D. These veterans' estimated annual savings ranged from $6 to $714, with an average savings of $353 per year (or 2% of their average annual income) by using Part D. CONCLUSIONS: Although VA pharmacy coverage has been deemed to be, on average, equivalent to Part D, some veterans living near poverty can reduce out of-pocket expenditures by using Medicare prescription coverage. Currently available data can identify veterans who are likely to achieve savings under Medicare. PMID- 17909388 TI - Developing indicators of inpatient adverse drug events through nonlinear analysis using administrative data. AB - BACKGROUND: Because of uniform availability, hospital administrative data are appealing for surveillance of adverse drug events (ADEs). Expert-generated surveillance rules that rely on the presence of International Classification of Diseases, 9th Revision Clinical Modification (ICD-9-CM) codes have limited accuracy. Rules based on nonlinear associations among all types of available administrative data may be more accurate. OBJECTIVES: By applying hierarchically optimal classification tree analysis (HOCTA) to administrative data, derive and validate surveillance rules for bleeding/anticoagulation problems and delirium/psychosis. RESEARCH DESIGN: Retrospective cohort design. SUBJECTS: A random sample of 3987 admissions drawn from all 41 Utah acute-care hospitals in 2001 and 2003. MEASURES: Professional nurse reviewers identified ADEs using implicit chart review. Pharmacists assigned Medical Dictionary for Regulatory Activities codes to ADE descriptions for identification of clinical groups of events. Hospitals provided patient demographic, admission, and ICD9-CM data. RESULTS: Incidence proportions were 0.8% for drug-induced bleeding/anticoagulation problems and 1.0% for drug-induced delirium/psychosis. The model for bleeding had very good discrimination and sensitivity at 0.87 and 86% and fair positive predictive value (PPV) at 12%. The model for delirium had excellent sensitivity at 94%, good discrimination at 0.83, but low PPV at 3%. Poisoning and adverse event codes designed for the targeted ADEs had low sensitivities and, when forced in, degraded model accuracy. CONCLUSIONS: Hierarchically optimal classification tree analysis is a promising method for rapidly developing clinically meaningful surveillance rules for administrative data. The resultant model for drug-induced bleeding and anticoagulation problems may be useful for retrospective ADE screening and rate estimation. PMID- 17909389 TI - Real-time vaccine safety surveillance for the early detection of adverse events. AB - BACKGROUND: Rare but serious adverse events associated with vaccines or drugs are often nearly impossible to detect in prelicensure studies and require monitoring after introduction of the agent in large populations. Sequential testing procedures are needed to detect vaccine or drug safety problems as soon as possible after introduction. OBJECTIVE: To develop and evaluate a new real-time surveillance system that uses dynamic data files and sequential analysis for early detection of adverse events after the introduction of new vaccines. RESEARCH DESIGN: The Centers for Disease Control and Prevention (CDC)-sponsored Vaccine Safety Datalink Project developed a real-time surveillance system and initiated its use in an ongoing study of a new meningococcal vaccine for adolescents. Dynamic data files from 8 health plans were updated and aggregated for analysis every week. The analysis used maximized sequential probability ratio testing (maxSPRT), a new signal detection method that supports continuous or time period analysis of data as they are collected. RESULTS: Using the new real-time surveillance system, ongoing analyses of meningococcal conjugate vaccine (MCV) safety are being conducted on a weekly basis. Two forms of maxSPRT were implemented: an analysis using concurrent matched controls, and an analysis based on expected counts of the outcomes of interest, which were estimated based on historical data. The analysis highlights both theoretical and operational issues, including how to (1) choose appropriate outcomes and stopping rules, (2) select control groups, and (3) accommodate variation in exposed:unexposed ratios between time periods and study sites. CONCLUSIONS: Real-time surveillance combining dynamic data files, aggregation of data, and sequential analysis methods offers a useful and highly adaptable approach to early detection of adverse events after the introduction of new vaccines. PMID- 17909390 TI - Medicare part D data: major changes on the horizon. AB - BACKGROUND: The 3 primary administrative data sets developed by the Centers for Medicare and Medicaid services (CMS) to support the Medicare Part D program implementation represent a valuable source of data for health services researchers. This paper describes the structure of the Medicare Part D program and the related databases, and discusses their utilization for research purposes. RESULTS: The Medicare Part D administrative data include information on plan benefits (integrated into the Health Plan Management System), beneficiary enrollment files, and prescription drug event (PDE) claims-type data. The enrollment data may be of use in investigating the benefits and plan types that appeal to beneficiaries, but their application is limited by the fact that, although individual beneficiaries' enrollment choices are recorded, only summary enrollment data are currently publicly available. PDE data are likely to be of most interest to researchers as they are detailed (including beneficiary identifiers, contract identifiers pharmacy provider information on drugs provided, drug cost, and insurance status), beneficiary-specific (allowing them to be linked to beneficiary characteristics), and an unusual output for a program reimbursed under a capitation-based system. Because PDE data are highly sensitive, only summary data on the number of Part D prescriptions filled are publicly available. CONCLUSIONS: Although the data collected in relation to the Medicare Part D program could be applied to many questions of interest to health services researchers, their utility is limited by the sensitive natures of many of these data, making it difficult currently to obtain access for research purposes. PMID- 17909391 TI - Evaluation and overview of the National Electronic Injury Surveillance System Cooperative Adverse Drug Event Surveillance Project (NEISS-CADES). AB - BACKGROUND: Adverse drug events (ADEs) are an important cause of patient injury. Although most medications are prescribed and used in the outpatient setting, prevention efforts focus on the inpatient setting, partly because of limited data on outpatient events. We describe and evaluate a new system for surveillance of outpatient ADEs treated in hospital emergency departments (EDs). METHODS: We used guidelines for evaluating public health surveillance systems, developed by the Centers for Disease Control and Prevention, to assess the performance of the National Electronic Injury Surveillance System-Cooperative Adverse Drug Event Surveillance project (NEISS-CADES) from January 1, 2004 through December 31, 2004. RESULTS: NEISS-CADES is a nationally representative surveillance system that identifies ADEs using ED clinical records. Of 10,383 reports in 2004, 100% listed patient age, sex, and disposition; 98% listed the implicated drugs. A 6 hospital evaluation of data quality, completeness, and other system attributes showed that NEISS-CADES data accurately reflected clinical records with respect to patient age and sex (100%), primary diagnosis (93%), implicated drugs (93%), primary treatments (80%), and diagnostic testing (61%). Sensitivity of case identification was estimated to be at least 0.33; estimated positive predictive value was 0.92. Data collection does not require additional work by clinical staff and has been well accepted by participating institutions. CONCLUSIONS: NEISS-CADES provides detailed and timely information on outpatient ADEs treated in EDs and identifies specific drugs and circumstances associated with these injuries. Findings from NEISS-CADES can help design and prioritize patient safety interventions for outpatient ADEs. PMID- 17909392 TI - Bronchial hyperresponsiveness in skiers: field test versus methacholine provocation? AB - INTRODUCTION: Asthma is frequently reported in endurance athletes, particularly in cross-country skiers. It has been reported that an exercise field test performed with the competitive type of exercise is the better for diagnosing asthma and bronchial hyperresponsiveness in athletes than bronchial provocation with methacholine. OBJECTIVE: The main objective was to compare an exercise field test consisting of a skiing competition with methacholine bronchial provocation in the diagnosis of asthma and bronchial hyperresponsiveness among skiers. METHODS: Twenty-four elite cross-country skiers from the Norwegian national teams (males/females = 16/8) were included in the study. The cumulative dose of inhaled methacholine causing a 20% fall in forced expiratory volume in 1 s (FEV1) (PD20) was compared with reduction in lung function (FEV1) >or= 10% from before to after an exercise field test consisting of a cross-country skiing competition, 10 km (males) and 7 km (females), respectively. RESULTS: Nine out of 24 (37.5%) athletes experienced a positive methacholine test (PD20 < 8 micromol) (2 females and 7 males), whereas only 2 of the 24 subjects (8.3%) had reductions in FEV1 >or= 10% after the exercise field test. A significant negative correlation was found between age and bronchial responsiveness, r = -0.47, P = 0.02. CONCLUSION: The methacholine bronchial provocation test is more sensitive than a sport specific exercise field test for identifying athletes with asthma and/or bronchial hyperresponsiveness. PMID- 17909393 TI - Cardiorespiratory fitness, BMI, and risk of hypertension: the HYPGENE study. AB - INTRODUCTION: Cardiorespiratory fitness and regular physical activity are inversely associated with the risk of hypertension, and exercise training has been shown to lower elevated blood pressure (BP). Genetic factors contribute significantly to the interindividual differences in endurance training-induced changes in BP. However, similar data on the genotype-by-fitness interactions on the risk of hypertension are scarce. METHODS: In 2000, we started a systematic collection of blood samples from all consenting subjects of the Aerobics Center Longitudinal Study (ACLS) with a goal to generate a resource for studies addressing genotype-by-fitness interaction effects on various health-related end points. Here, we introduce the rationale and design of the first study based on the ACLS genetics resource focusing on hypertension as the health outcome (HYPGENE study), and we report the associations of cardiorespiratory fitness and body mass index (BMI) with the risk of hypertension. All HYPGENE subjects (N = 1234) were healthy and normotensive at their first clinic visit. Cases (N = 629) developed hypertension during the follow-up period (mean 8.7 yr), whereas controls (N = 605) remained normotensive (mean follow-up 10.1 yr). RESULTS: Cardiorespiratory fitness was the strongest predictor of the hypertension risk, with each maximal metabolic equivalent unit being associated with a 19% lower risk (95% confidence interval [95% CI], 12-24%). Each baseline BMI unit was associated with a 9% higher hypertension risk (95% CI, 4-13%). However, the association of BMI was greatly attenuated (odds ratio 1.04 [95% CI, 0.99-1.09]) when fitness also was included in the model. CONCLUSIONS: The HYPGENE study will provide an excellent resource to address hypotheses regarding the genetic basis of hypertension while taking cardiorespiratory fitness level into account. PMID- 17909394 TI - Physical inactivity, depression, and risk of cardiovascular mortality. AB - PURPOSE: Studies indicate that depression may increase risk of cardiovascular disease (CVD) in addition to classical risk factors. One of the hypotheses to explain this relation is that depressed subjects become physically inactive. We set out to determine the role of physical inactivity in the relation between depressive symptoms and cardiovascular mortality. METHODS: Data were used from the population-based prospective Finland, Italy, and the Netherlands Elderly (FINE) Study. Depressive symptoms were measured with the Zung Self-Rating Depression Scale in 909 elderly men, aged 70-90 yr, free of CVD and diabetes at baseline in 1990. Physical activity was assessed with a questionnaire for retired men. Hazard ratios (HR) for 10-yr cardiovascular mortality were calculated, adjusting for demographics and cardiovascular risk factors. RESULTS: At baseline, men with more depressive symptoms were less physically active (722 min.wk; 95% confidence interval (CI), 642-802) than men with few depressive symptoms (919 min.wk; 95% CI, 823-1015). During 10 yr of follow-up, 256 (28%) men died from CVD. The adjusted HR of cardiovascular mortality for a decrease of 30 min.d in physical activity was 1.09 (95% CI, 1.04-1.14). An increase in depressive symptoms with one standard deviation was associated with a higher cardiovascular mortality risk (HR = 1.42; 95% CI, 1.26-1.60). After additional adjustment for physical activity the risk decreased (9%), but an independent risk remained (HR = 1.37; 95% CI, 1.21-1.56). The excess risk on cardiovascular mortality attributable to the combined effect of depressive symptoms with inactivity was 1.47 (95% CI, -0.17 to 3.11). CONCLUSIONS: In the present study, the increased risk of depressive symptoms on cardiovascular mortality could not be explained by physical inactivity. However, our results suggest that depressive symptoms and physical inactivity may interact to increase cardiovascular mortality risk. PMID- 17909395 TI - Physical activity patterns and their correlates among Chinese men in Shanghai. AB - INTRODUCTION: Physical activity (PA) is inversely related to the risk of many chronic diseases. Understanding PA patterns and their correlates thus has significant public health implications. METHODS: We evaluated PA patterns and their association with socioeconomic status and lifestyle factors in the Shanghai Men's Health Study (SMHS), a cohort of 61,582 Chinese men (participation rate: 74.1%) ages 40-74 living in eight communities of urban Shanghai, China. Information on PA from exercise, household chores, and walking and bicycling for transportation and daily living activities was collected by in-person interviews using a validated questionnaire. Logistic regression analyses were conducted. RESULTS: Participation in exercise was reported by 35.6% of study participants, walking and cycling for transportation by 22.6% and 23.5%, and walking and cycling for daily living activities by 99.9% and 24.5%. Nine percent had high-PA jobs. All kinds of PA, except household chores, were more common in older men. Education and income levels were positively associated with exercise and housework but inversely associated with transportation and daily living activities. Men with higher BMI participated in more exercise, whereas those with higher waist-to-hip ratio (WHR) were less active in all kinds of PA. Current smokers, particularly heavy smokers, were less active in all kinds of PA compared with former smokers and nonsmokers. Current alcohol drinkers, tea drinkers, and ginseng users were more likely to participate in exercise but less likely to participate in nonexercise PA. Total energy intake was positively associated with PA, except for household chores. CONCLUSIONS: Despite low participation in exercise/sports, most middle-aged and elderly Chinese men in Shanghai participate in a high level of nonexercise PA. Their PA patterns are closely associated with socioeconomic/lifestyle factors. PMID- 17909396 TI - Effects of exercise load and blood-flow restriction on skeletal muscle function. AB - Resistance training at low loads with blood flow restriction (BFR) (also known as Kaatsu) has been shown to stimulate increases in muscle size and strength. It is unclear how occlusion pressure, exercise intensity, and occlusion duration interact, or which combination of these factors results in the most potent muscle stimulus. PURPOSE: To determine the effect of eight BFR protocols on muscle fatigue (decrement in maximal voluntary contraction (MVC) after the performance of exercise), and to compare the decrement in MVC with the currently recommended resistance exercise intensity (~80% MVC). METHODS: During five test sessions, 21 subjects (14 males and 7 females, 27.7 +/- 4.9 yr) completed nine protocols, each consisting of three sets of knee extensions (KE) to failure. One protocol was high-load (HL) exercise (80% MVC) with no BFR, and the other eight were BFR at varying levels of contraction intensity (20 or 40% MVC), occlusion pressure (partial (~160 mm Hg) or complete (~300 mm Hg)), and occlusion duration (off during the rest between sets or continuously applied). To evaluate each protocol, MVC were performed before and after exercise, and the decrement in force was calculated. RESULTS: Three sets of KE at 20% MVC with continuous partial occlusion (20%(ConPar)) resulted in a greater decrement in MVC compared with HL (31 vs 19%, P = 0.001). None of the other BFR protocols were different from the HL protocol, nor were they different from 20%(ConPar) (P > 0.05). CONCLUSION: All BFR protocols elicited at least as much fatigue as HL, even though lower loads were used. The 20%(ConPar) protocol was the only one that elicited significantly more fatigue than HL. Future research should evaluate protocol training effectiveness and overall safety of BFR exercise. PMID- 17909397 TI - The influence of exercise training on inflammatory cytokines and C-reactive protein. AB - PURPOSE: The purpose of this study was to examine the influence of a 12-wk exercise training program on inflammatory cytokine and C-reactive protein (CRP) concentrations. A secondary purpose was to determine whether training-induced changes in cytokines and CRP were influenced by age. METHODS: Twenty-nine younger (18-35 yr) and 31 older (65-85 yr) subjects were assigned to young physically active (YPA, N = 15; 25 +/- 5 yr), young physically inactive (YPI, N= 14; 25 +/- 4.7 yr), old physically active (OPA, N = 14; 71 +/- 4 yr), or old physically inactive (OPI, N = 17; 71 +/- 4 yr) groups. The inactive groups completed 12 wk (3 d.wk) of aerobic and resistance exercises, and the physically active control groups continued their normal exercise programs. Blood samples were collected before and after the 12-wk period, and the concentrations of serum CRP, plasma interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and interleukin-1 beta (IL-1beta) were determined using separate ELISA. RESULTS: Control (YPA and OPA) estimated VO2max was unchanged. Exercise training increased estimated VO2max an average of 10.4% and increased strength by an average of 38.1% in both PI groups. Serum CRP decreased with training (YPI and OPI) groups and was not different from the YPA and OPA groups after training. Plasma IL-6 and IL-1beta did not change, whereas TNF-alpha was higher than YPI and YPA at baseline and after the intervention period. CONCLUSION: These results support the use of combined aerobic/resistance training as a modality to reduce the risk of cardiovascular disease development as defined by a decrease in serum CRP concentration in healthy humans. PMID- 17909398 TI - Separate and combined effects of airflow and rehydration during exercise in the heat. AB - PURPOSE: To determine whether airflow is required to obtain the beneficial effects of rehydration (thermoregulatory and cardiovascular) during exercise in dry heat. METHODS: Ten moderately trained (VO2max = 55 +/- 8 mL.kg(-1).min(-1)) heat acclimated males pedaled for 60 min at 60% VO2max in a hot-dry environment (36 +/- 1 degrees C; 29 +/- 2% relative humidity) on four different occasions: 1) without rehydration or forced airflow (control trial; CON); 2) rehydrating 100% of sweat losses by ingestion of a 6% carbohydrate-electrolyte solution (rehydration trial; REH); 3) receiving airflow at a velocity of 2.55 m.s(-1) (wind trial; WIND); and 4) combining airflow and rehydration (W + R). RESULTS: Without airflow, rehydration alone (REH) did not lower rectal temperature below CON (39.0 +/- 0.1 vs 39.1 +/- 0.1 degrees C at 60 min; respectively). However, with airflow, rehydration reduced final rectal temperature (38.8 +/- 0.1 vs 38.5 +/- 0.1 degrees C; P < 0.05; WIND vs W + R). In the trials with wind (WIND and W + R), skin temperature was reduced by about 0.6 degrees C (P < 0.05), and heart rate drift was prevented. In the trials with rehydration (REH and W + R trials), cardiac output (CO2-rebreathing technique) was maintained higher than CON (16.5 +/- 0.4 and 17.0 +/- 0.7 vs 15.4 +/- 0.4 L.min(-1), respectively; P < 0.05). CONCLUSION: When exercising in a hot-dry environment, airflow is required for rehydration to improve thermoregulation and cardiovascular function. PMID- 17909400 TI - Regular physical activity influences plasma ghrelin concentration in adolescent girls. AB - PURPOSE: We examined the effect of regular physical activity on plasma ghrelin concentration after onset of puberty in girls. In addition, we also examined the association of fasting plasma ghrelin concentration with various plasma biochemical, body composition, and aerobic capacity variables in healthy adolescent girls. METHOD: Fifty healthy schoolgirls ages 11 to 16 yr were divided either into a physically active (N = 25) or a physically inactive (N = 25) group. The physically active group consisted of swimmers who had trained on an average of 6.2 +/- 2.0 h.wk(-1) for the last 2 yr, whereas the inclusion criterion for the physically inactive group was the participation in physical education classes only. The subjects were matched for age (+/- 1 yr) and body mass index (BMI; +/- 2 kg.m(-2)). Maturation I group (14 matched pairs) included pubertal stages 2 and 3, and maturation II group (11 matched pairs) included pubertal stages 4 and 5. RESULTS: Physically active girls had significantly higher (P < 0.05) mean plasma ghrelin levels than the physically inactive girls (maturation I: 1152.1 +/- 312.9 vs 877.7 +/- 114.8 pg.mL(-1); maturation II: 1084.0 +/- 252.5 vs 793.4 +/- 164.9 pg.mL(-1)). Plasma ghrelin concentration was negatively related to percent body fat, fat mass, peak oxygen consumption per kilogram of body mass, leptin, estradiol, insulin, and insulin-like growth factor-I (IGF-I) (r > -0.298; P < 0.05). Multivariate linear regression analysis to determine the predictors of ghrelin concentration using the variables that were significantly associated with ghrelin concentration demonstrated that plasma IGF-I was the most important predictor of plasma ghrelin concentration (beta = -0.396; P = 0.008). CONCLUSION: Regular physical activity influences plasma ghrelin concentrations in girls with different pubertal maturation levels. Plasma IGF-I concentration seems to be the main determinant of circulating ghrelin in healthy, normal-weight adolescent girls. PMID- 17909399 TI - GXT responses in altitude-acclimatized cyclists during sea-level simulation. AB - PURPOSE: This study examined the effects of gender on graded exercise stress test (GXT) response in moderate-altitude (MA)-acclimatized cyclists during sea-level (SL) simulation. It was hypothesized that alterations in arterial saturation would relate to changes in VO2peak. METHODS: Twenty competitive cyclists (12 males, 8 females) who were residents of MA locations underwent two randomized bicycle GXTs: one under local normoxic hypobaria, and the other under simulated SL conditions. RESULTS: Under the SL condition, the cyclists demonstrated a significant increase (2-3%) in absolute and relative VO2peak, improved (4%) economy at lactate threshold (LT), and time-adjusted peak power (7%); the range of improvement between individuals varied from -6% to +25%. Simulated SL also resulted in a greater arterial saturation (S(a)O2) at rest and VO2peak, and significantly less desaturation (4 vs 8%) from rest to VO2peak. The individual variability in the change (Delta) in VO2peak was not significantly correlated to SL S(a)O2 or any other S(a)O2 variable analyzed, regardless of whether we examined each gender individually or combined. Significant correlations were found between Delta-peak power and Delta-economy as well as Delta-VO2peak and Delta-GXT time. These correlations as well as degree of improvement varied by gender. CONCLUSIONS: These data suggest that chronic residence at MA may attenuate the occurrence of exercise-induced arterial hypoxemia and eliminate the relationship between S(a)O2 and Delta-VO2peak that has been reported among SL residents acutely exposed to altitude. Additionally, the improvements that occur in predictors of aerobic performance when MA residents are exposed acutely to SL conditions have a large degree of individual variability, and the mechanism(s) for improvement may vary by gender. PMID- 17909401 TI - Detection of darbepoetin alfa misuse in urine and blood: a preliminary investigation. AB - INTRODUCTION: Darbepoetin alfa is a modified erythropoietin (EPO) molecule with a longer serum half-life than recombinant human erythropoietin (rhEPO). Because the detection period of rhEPO in urine is only 2-3 d after the last injection, blood algorithms have been developed in order to expand the detection window of rhEPO misuse. The main objectives were to establish the period of detection of darbepoetin alfa by isoelectric focusing (IEF) and examine the applicability of blood algorithms and individual variations in blood variables in an antidoping context. METHODS: Six recreationally active males and six recreationally active females had 0.78 microg.kg(-1).wk(-1) of darbepoetin alfa administered for 3 wk. Blood and urine samples were collected continuously during and after administration. Urine samples were analyzed by IEF and immunoblotting for darbepoetin alfa, and blood samples were analyzed for erythropoietic sensitive blood variables on a hematological analyzer. RESULTS: Darbepoetin alfa was detected in 8 of 12 samples at 10 d after the last injection. Ten subjects showed variations in hemoglobin concentration [Hb] > 10%, whereas only three males and one female exceeded suggested upper [Hb] limits of 17.0 and 16.0 g.dL(-1), respectively. Four subjects exceeded the 1:1000 ON- as well as the OFF-model cutoff limit. CONCLUSION: The large number of samples containing detectable amounts of darbepoetin alfa at 10 d into the washout period stipulate the possibility of a 7-d window of detection after administration, wherein a sample would be regarded as an adverse analytical finding. The marked variations in all examined blood parameters could be used for the targeting of urine samples. These preliminary findings open up for larger scale studies with more frequent urine sampling in the washout period on elite athletes. PMID- 17909402 TI - Hb mass measurement suitable to screen for illicit autologous blood transfusions. AB - PURPOSE: An increase of hemoglobin (Hb) mass is the key target of blood doping practices to enhance performance as it is a main determinant of maximal oxygen uptake. Although detection methods exist for doping with recombinant EPO and homologous blood transfusions, autologous transfusions remain virtually undetectable. In this context, the most sensitive parameter would be a determination of Hb mass itself. The purpose therefore was to establish whether Hb mass measurements by the optimized CO-rebreathing method allow screening for the withdrawal and reinfusion of autologous red blood cells. METHODS: The optimized CO-rebreathing method was used for evaluation of Hb mass in two groups at three time points (duplicate measurements: 1) baseline, 2) after donation, and 3) after reinfusion). Group I (N = 6) was to donate and receive 1 unit of packed red cells (PRC) in contrast to two PRC in group II (N = 4). The time span between withdrawal and reinfusion was 2 d. RESULTS: The mean Hb content of the blood units was 59.0 +/- 3.9 g (group I) and 108.3 +/- 1.3 g (group II). Hb mass decreased significantly after blood withdrawal (-89 +/- 16 g in group I and -120 +/- 14 g in group II) and increased significantly after reinfusion (group I: 70 +/- 16 g; group II: 90 +/- 9 g) but was lower than at baseline (group I: -19 +/- 17 g; group II: -30 +/- 14 g). The total error of measurements for the duplicate measures ranged between 0.8 and 3.1% (Hb mass: 6.4-22.1 g). CONCLUSION: Hb mass determination with the optimized CO-rebreathing method has sufficient precision to detect the absolute differences in Hb mass induced by blood withdrawal and autologous reinfusion. Thus, it may be suited to screen for artificially induced alterations in Hb mass. PMID- 17909403 TI - Positive and negative loading and mechanical output in maximum vertical jumping. AB - PURPOSE: The aim of this study was to evaluate the effect of external loading on mechanics of vertical jumping. We hypothesized that the muscular mechanical output could be higher under no-load conditions than in the presence of either positive or negative external loads. METHODS: Fifteen physically active men performed maximal countermovement jumps (CMJ) on a force plate while a pulley system provided approximately constant vertical force acting in a way to either reduce or increase the body weight. As a result, the weight of the body approximately corresponded to the gravity acceleration from 0.70 to 1.30 g (g = 9.81 m.s(-2)). RESULTS: Regarding the jumping kinematics, we observed a significant (P < 0.001) load-associated decrease in both the peak velocity and lowering of the center of mass during the eccentric jump phase, but not in the duration of the subsequent concentric jump phase. Regarding the muscular mechanical output, both the mean power (P ) and peak momentum (M) revealed significant (P < 0.001) changes associated with loading, and further post hoc analyses revealed significantly higher values (P < 0.05-0.001) of both P and M for 1.00 g compared with most of the other loading conditions applied. CONCLUSION: The results suggest that subject's own body provides the optimal load for producing maximum mechanical output in vertical jumping. If corroborated by the results of future studies performed on other rapid movement, our findings could support the hypothesis that the muscular system is designed for producing maximum mechanical output in rapid movements when loaded only with the weight and inertia of its own body. PMID- 17909404 TI - The effect of technique change on knee loads during sidestep cutting. AB - PURPOSE: To identify the effect of modifying sidestep cutting technique on knee loads and predict what impact such change would have on the risk of noncontact anterior cruciate ligament injury. METHODS: A force platform and motion-analysis system were used to record ground-reaction forces and track the trajectories of markers on 15 healthy males performing sidestep cutting tasks using their normal technique and nine different imposed techniques. A kinematic and inverse dynamic model was used to calculate the three-dimensional knee postures and moments. RESULTS: The imposed techniques of foot wide and torso leaning in the opposite direction to the cut resulted in increased peak valgus moments experienced in weight acceptance. Higher peak internal rotation moments were found for the foot wide and torso rotation in the opposite direction to the cut techniques. The foot rotated in technique resulted in lower mean flexion/extension moments, whereas the foot wide condition resulted in higher mean flexion/extension moments. The flexed knee, torso rotated in the opposite direction to the cut and torso leaning in the same direction as the cut techniques had significantly more knee flexion at heel strike. CONCLUSION: Sidestep cutting technique had a significant effect on loads experienced at the knee. The techniques that produced higher valgus and internal rotation moments at the knee, such as foot wide, torso leaning in the opposite direction to the cut and torso rotating in the opposite direction to the cut, may place an athlete at higher risk of injury because these knee loads have been shown to increase the strain on the anterior cruciate ligament. Training athletes to avoid such body positions may result in a reduced risk of noncontact anterior cruciate ligament injures. PMID- 17909405 TI - Modulation of prelanding lower-limb muscle responses in athletes with multiple ankle sprains. AB - PURPOSE: The objective of this study was to investigate modulation in prelanding muscle responses and its associated impact force on landing from unexpected and self-initiated drops in male basketball players with a history of bilateral multiple ankle sprains (BMAS). METHODS: Prelanding EMG responses were recorded in four lower-limb muscles, together with the impact force on landing, while 20 healthy and 19 basketball players with BMAS performed unexpected, self-initiated drops from a height of 30 cm. RESULTS: Group differences were detected after self initiated but not unexpected drops. Two main changes in prelanding EMG responses were observed in the injured basketball players during the self-initiated drops. First, tibialis anterior (TA) was activated significantly earlier in the injured group, whereas left tensor fascia latae appeared closer to the moment of landing (P < 0.025) than in the healthy players. Second, cocontraction indexes between left TA and peroneus longus, and left TA and medial gastrocnemius, were significantly greater in the injured than in the healthy players (P < 0.025). On landing, higher magnitude-of-impact forces were observed in the injured players on the right leg (by 23%, P = 0.012). CONCLUSION: In basketball players with BMAS, modulation of prelanding muscle response latencies occurred in injured (ankle) and uninjured (hip) joints during self-initiated but not unexpected drops. Greater cocontraction index between the left ankle muscle pairs in preparation for landing from self-initiated drops, and a significantly higher magnitude of impact force in the right leg on landing, were observed in the injured players. PMID- 17909406 TI - Kinematic changes during a 100-m front crawl: effects of performance level and gender. AB - PURPOSE: This study analyzed kinematic changes during a 100-m front crawl to investigate the effects of performance level and gender, comparing 12 high-speed males, 8 medium-speed males, 8 low-speed males, and 8 high-speed females. METHODS: Assessments were made throughout the race in a 25-m pool divided into five zones of 5 m. Velocity (V), stroke rate (SR), and stroke length (SL) were calculated for each 25-m length (L1 to L4) and for each 5-m zone. Four stroke phases were identified by video analysis, and the index of coordination (IdC) was calculated. Three modes of arm coordination were identified: catch-up, opposition, and superposition. The leg kick was also analyzed. RESULTS: The high speed male swimmers were distinguished by higher V (1.89 m.s(-1)), SR (0.78 Hz), SL (2.16 m per stroke), propulsive phase (54%), and IdC (3.8%) (P < 0.05), and by the stability of these values throughout the race. The medium- and low-speed males had an opposition coordination (-1% < IdC < 1%) during the third length of the 100 m. Because of fatigue in length 4, they spent more time with the hand in the push phase (possibly because of a decrease in hand velocity) and changed to superposition coordination (medium-speed males: IdC = 2.78%; low-speed males: IdC = 1.12%) (P < 0.05). This change was ineffective, however, as SL continued to decrease throughout the 100 m (P < 0.05). The main gender findings were the greater SL of the males versus the females (1.81 m per stroke) (P < 0.05) and the similar IdC of both high-speed groups (females: 4.4%). CONCLUSION: The high-speed swimmers were characterized by higher and more stable SL and IdC. The principal gender effect was greater SL in the males than in the females. PMID- 17909407 TI - Vibration exposure and biodynamic responses during whole-body vibration training. AB - PURPOSE: Excessive, chronic whole-body vibration (WBV) has a number of negative side effects on the human body, including disorders of the skeletal, digestive, reproductive, visual, and vestibular systems. Whole-body vibration training (WBVT) is intentional exposure to WBV to increase leg muscle strength, bone mineral density, health-related quality of life, and decrease back pain. The purpose of this study was to quantitatively evaluate vibration exposure and biodynamic responses during typical WBVT regimens. METHODS: Healthy men and women (N = 16) were recruited to perform slow, unloaded squats during WBVT (30 Hz; 4 mm(p-p)), during which knee flexion angle (KA), mechanical impedance, head acceleration (Ha(rms)), and estimated vibration dose value (eVDV) were measured. WBVT was repeated using two forms of vibration: 1) vertical forces to both feet simultaneously (VV), and 2) upward forces to only one foot at a time (RV). RESULTS: Mechanical impedance varied inversely with KA during RV (effect size, eta(p)(2): 0.668, P < 0.01) and VV (eta(p)(2): 0.533, P < 0.05). Ha(rms) varied with KA (eta(p)(2): 0.686, P < 0.01) and is greater during VV than during RV at all KA (P < 0.01). The effect of KA on Ha(rms) is different for RV and VV (eta(p)(2): 0.567, P < 0.05). The eVDV associated with typical RV and VV training regimens (30 Hz, 4 mm(p-p), 10 min.d(-1)) exceeds the recommended daily vibration exposure as defined by ISO 2631-1 (P < 0.01). CONCLUSIONS: ISO standards indicate that 10 min.d(-1) WBVT is potentially harmful to the human body; the risk of adverse health effects may be lower during RV than VV and at half-squats rather than full-squats or upright stance. More research is needed to explore the long term health hazards of WBVT. PMID- 17909408 TI - Effects of plyometric and weight training on muscle-tendon complex and jump performance. AB - PURPOSE: The purpose of this study was to investigate the effects of plyometric and weight training protocols on the mechanical properties of muscle-tendon complex and muscle activities and performances during jumping. METHODS: Ten subjects completed 12 wk (4 d.wk(-1)) of a unilateral training program for plantar flexors. They performed plyometric training on one side (PT; hopping and drop jump using 40% of 1RM) and weight training on the other side (WT; 80% of 1RM). Tendon stiffness was measured using ultrasonography during isometric plantar flexion. Three kinds of unilateral jump heights using only ankle joint (squat jump: SJ; countermovement jump: CMJ; drop jump: DJ) on sledge apparatus were measured. During jumping, electromyographic activities were recorded from plantar flexors and tibial anterior muscle. Joint stiffness was calculated as the change in joint torque divided by the change in ankle angle during eccentric phase of DJ. RESULTS: Tendon stiffness increased significantly for WT, but not for PT. Conversely, joint stiffness increased significantly for PT, but not for WT. Whereas PT increased significantly jump heights of SJ, CMJ, and DJ, WT increased SJ only. The relative increases in jump heights were significantly greater for PT than for WT. However, there were no significant differences between PT and WT in the changes in the electromyographic activities of measured muscles during jumping. CONCLUSION: These results indicate that the jump performance gains after plyometric training are attributed to changes in the mechanical properties of muscle-tendon complex, rather than to the muscle activation strategies. PMID- 17909409 TI - Oxygen-uptake efficiency slope as a determinant of fitness in overweight adolescents. AB - PURPOSE: Peak oxygen uptake (VO2peak) is frequently difficult to assess in overweight individuals; therefore, submaximal measures that predict VO2peak are proposed as substitutes. Oxygen uptake efficiency slope (OUES) has been suggested as a submaximal measurement of cardiorespiratory fitness that is independent of exercise intensity. There are few data examining its value as a predictor of V O2peak in severely overweight adolescents. METHODS: One hundred seven severely overweight (BMI Z 2.50 +/- 0.34) and 43 nonoverweight (BMI Z 0.13 +/- 0.84) adolescents, performed a maximal cycle ergometer test with respiratory gas exchange measurements. OUES was calculated through three exercise intensities: lactate inflection point (OUES LI), 150% of lactate inflection point (OUES 150), and VO2peak (OUES PEAK). RESULTS: When adjusted for lean body mass, VO2peak and OUES at all exercise intensities were lower in overweight subjects (VO2peak: 35.3 +/- 6.4 vs 46.8 +/- 7.9 mL.kg(-1) LBM.min(-1), P < 0.001; OUES LI: 37.9 +/- 10.0 vs 43.7 +/- 9.2 mL.kg(-1) LBM.min(-1).logL(-1) P < 0.001; OUES 150: 41.6 +/- 9.0 vs 49.8 +/- 11.1 mL.kg(-1) LBM.min(-1).logL(-1) P < 0.001; and OUES PEAK: 45.1 +/ 8.7 vs 52.8 +/- 9.6 mL.kg(-1) LBM.min(-1).logL(-1) P < 0.001). There was a significant increase in OUES with increasing exercise intensity in both groups (P < 0.001). OUES at all exercise intensities was a significant predictor of VO2peak for both groups (r2 = 0.35-0.83, P < 0.0001). However, limits of agreement for predicted VO2peak relative to actual VO2peak were wide (+/- 478 to +/- 670 mL.min(-1)). CONCLUSIONS: OUES differs significantly in overweight and nonoverweight adolescents. The wide interindividual variation and the exercise intensity dependence of OUES preclude its use in clinical practice as a predictor of VO2peak. PMID- 17909410 TI - Effect of hydration state on strength, power, and resistance exercise performance. AB - PURPOSE: Although many studies have attempted to examine the effect of hypohydration on strength, power, and high-intensity endurance, few have successfully isolated changes in total body water from other variables that alter performance (e.g., increased core temperature), and none have documented the influence of hypohydration on an isotonic, multiset, multirepetition exercise bout typical of resistance exercise training. Further, no investigations document the effect of hypohydration on the ability of the central nervous system to stimulate the musculature, despite numerous scientists suggesting this possibility. The purposes of this study were to examine the isolated effect of hydration state on 1) strength, power, and the performance of acute resistance exercise, and 2) central activation ratio (CAR). METHODS: Seven healthy resistance-trained males (age = 23 +/- 4 yr, body mass = 87.8 +/- 6.8 kg, body fat = 11.5 +/- 5.2%) completed three resistance exercise bouts in different hydration states: euhydrated (EU), hypohydrated by approximately 2.5% body mass (HY25), and hypohydrated by approximately 5.0% body mass (HY50). Investigators manipulated hydration status via exercise-heat stress and controlled fluid intake 1 d preceding testing. RESULTS: Body mass decreased 2.4 +/- 0.4 and 4.8 +/- 0.4% during HY25 and HY50, respectively. No significant differences existed among trials in vertical jump height, peak lower-body power (assessed via jump squat), or peak lower-body force (assessed via isometric back squat). CAR tended to decrease as hypohydration increased (EU = 95.6 +/- 4.9%, HY25 = 94.0 +/- 3.1%, HY50 = 92.5 +/- 5.1%; P = 0.075, eta(p)(2) = 0.41). When evaluated as a function of the percentage of total work completed during a six-set back squat protocol, hypohydration significantly decreased resistance exercise performance during sets 2-3 and 2-5 for HY25 and HY50, respectively. CONCLUSION: These data indicate that hypohydration attenuates resistance exercise performance; the role of central drive as the causative mechanism driving these responses merits further research. PMID- 17909411 TI - Chronic static stretching improves exercise performance. AB - PURPOSE: This study investigated the influence of static stretching exercises on specific exercise performances. METHODS: Thirty-eight volunteers participated in this study. The stretching group (STR) consisted of 8 males and 11 females whose activity was limited to a 10-wk, 40-min, 3-d.wk(-1) static stretching routine designed to stretch all the major muscle groups in the lower extremity. The control group (CON) consisted of 8 males and 11 females who did not participate in any kind of regular exercise routine during the study. Each subject was measured before and after for flexibility, power (20-m sprint, standing long jump, vertical jump), strength (knee flexion and knee extension one-repetition maximum (1RM)), and strength endurance (number of repetitions at 60% of 1RM for both knee flexion and knee extension). RESULTS: STR had significant average improvements (P < 0.05) for flexibility (18.1%), standing long jump (2.3%), vertical jump (6.7%), 20-m sprint (1.3%), knee flexion 1RM (15.3%), knee extension 1RM (32.4%), knee flexion endurance (30.4%) and knee extension endurance (28.5%). The control group showed no improvement. CONCLUSION: This study suggests that chronic static stretching exercises by themselves can improve specific exercise performances. It is possible that persons who are unable to participate in traditional strength training activities may be able to experience gains through stretching, which would allow them to transition into a more traditional exercise regimen. PMID- 17909412 TI - Empirical evaluation of physical activity recommendations for weight control in women. AB - PURPOSE: Recent recommendations advise 30-60 min of physical activity per day to prevent weight gain and 60-90 min to prevent weight regain. No studies have used objective measures of physical activity to verify these public health recommendations. The purpose of this study was to use objective measures to quantify the amount and intensity of physical activity in a weight-loss maintainer group and an always-normal-weight group, and, thus,empirically evaluate the recommendations for prevention of weight gain versus regain. METHODS: The weight-loss-maintainer group (N = 135) lost >or= 30.6 kg, maintained >or= 10% weight loss for 14.2 yr, and had a BMI of 22.0 kg.m(-2). The always normal-weight group (N = 102) had a BMI of 21.1 kg.m(-2) and no history of overweight. Accelerometry was used to assess the amount and intensity of physical activity. RESULTS: The weight-loss-maintainer group spent significantly more minutes per day than the always-normal-weight group in physical activity (58.6 vs 52.1; P = 0.0001), largely because of more time spent in higher-intensity activities (24.4 vs 16.9; P = 0.02). The majority of individuals in the always normal-weight group engaged in 30-60 min.d(-1) of physical activity, whereas a greater proportion of individuals in the weight-loss-maintainer group engaged in > 60 min (P = 0.002). CONCLUSIONS: Findings support current recommendations that more activity may be needed to prevent weight regain than to prevent weight gain. Including some higher-intensity activity may also be advisable for weight-loss maintenance. PMID- 17909413 TI - A comparative analysis of pedometry in measuring physical activity of children. AB - PURPOSE: The purpose of this study was to examine the step and physical activity time output features of the Walk4Life LS2505 pedometer under field physical activity conditions. METHODS: Data were collected on 288 (12.62 +/- 1.23 yr) participants during a school-based structured physical activity program. Participants' physical activity levels were concurrently measured via the Yamax SW701 (Yamax Corp., Japan) and Walk4Life LS2505 (Walk4Life Inc., Plainfield, IL) pedometers, and System for Observing Fitness Instruction Time (SOFIT) physical activity coding mechanism. Relative and absolute agreement between SW701 (criterion) and LS2505 steps per minute, and SOFIT (criterion) and LS2505 physical activity time (min) were analyzed overall, and across physical activity content themes and physical activity quartiles. RESULTS: Physical activity measure correlations were moderately strong to strong (r = 0.85-0.98, P < 0.05); however, the LS2505 significantly underestimated steps per minute (M(diff) = 6.37 +/- 5.79, P < 0.05) and overestimated physical activity time (M(diff) = -7.73 +/- 3.13, P < 0.05). When LS2505 steps per minute were examined across physical activity themes and quartiles, clinically acceptable absolute error scores (or= 0.05). Both lanyard and waistband units significantly overestimated distance traveled during walking trials (P or= 0.05). The relative technical error of measurement (TEM) of the raw data ranged from 3.74 to 15.51%, and average absolute errors ranged from 5.03 to 8.53% for all trials. A significant position by AT mode interaction was observed for clean data (P < 0.05). Relative TEM for the clean data ranged from 1.42 to 1.98%, and average absolute errors ranged from 0.32 to 1.97%. Intraclass correlations (ICC) were poor to fair for all trials using raw and cleaned data. CONCLUSION: Signal noise during unit initialization may adversely affect unit performance; however, application of data-cleaning procedures to remove data associated with signal noise improves unit ability to measure distance. Results suggest that the lanyard position is the optimal placement for units during data collection. PMID- 17909416 TI - Lapses and psychosocial factors related to physical activity in early postmenopause. AB - PURPOSE: After menopause, leisure physical activity (PA) levels seem to decline for reasons that are not completely understood. This study examines the associations between PA, lapses in PA, and psychosocial factors in early postmenopausal women. METHODS: This cross-sectional analysis included 497 women from the Women on the Move through Activity and Nutrition study. PA was assessed with a past-year, interviewer-administered Modifiable Activity Questionnaire. Measures of activity lapses of >or= 2 wk in the past 6 months, exercise decision making, processes of change, and self-efficacy were collected along with Beck Depression Inventory, State-Trait Anxiety Inventory, Cohen Perceived Stress Scale, and Short Form-36. RESULTS: Mean age of participants was 56.9 yr. Compared with less active women, women with significantly higher activity levels reported greater exercise self-efficacy (r = 0.31), more frequent use of behavioral exercise processes of change (r = 0.31), greater perceived benefits for PA (r = 0.22), and better physical quality of life (r = 0.16) (all P < 0.001). Women reporting no activity lapses had higher reported activity levels than regularly active women with lapses or occasionally active women with lapses (P < 0.0001 for trend). Of the women who reported lapses, 24% reported low self-confidence, 43% reported difficulty controlling their weight, and 55% reported difficulty maintaining their diet when they lapsed from PA. Thirty-nine percent of women reporting lapses did not resume PA (i.e., relapsed to inactivity). Higher anxiety and depressive symptoms, and less frequent use of behavioral exercise processes of change, were associated with relapse to inactivity. CONCLUSIONS: Future interventions for early postmenopausal women should consider psychosocial factors when attempting to encourage and maintain higher levels of PA. Addressing and preventing PA lapses may help to achieve PA goals in this population. PMID- 17909417 TI - American College of Sports Medicine position stand. The female athlete triad. AB - The female athlete triad (Triad) refers to the interrelationships among energy availability, menstrual function, and bone mineral density, which may have clinical manifestations including eating disorders, functional hypothalamic amenorrhea, and osteoporosis. With proper nutrition, these same relationships promote robust health. Athletes are distributed along a spectrum between health and disease, and those at the pathological end may not exhibit all these clinical conditions simultaneously. Energy availability is defined as dietary energy intake minus exercise energy expenditure. Low energy availability appears to be the factor that impairs reproductive and skeletal health in the Triad, and it may be inadvertent, intentional, or psychopathological. Most effects appear to occur below an energy availability of 30 kcal.kg(-1) of fat-free mass per day. Restrictive eating behaviors practiced by girls and women in sports or physical activities that emphasize leanness are of special concern. For prevention and early intervention, education of athletes, parents, coaches, trainers, judges, and administrators is a priority. Athletes should be assessed for the Triad at the preparticipation physical and/or annual health screening exam, and whenever an athlete presents with any of the Triad's clinical conditions. Sport administrators should also consider rule changes to discourage unhealthy weight loss practices. A multidisciplinary treatment team should include a physician or other health-care professional, a registered dietitian, and, for athletes with eating disorders, a mental health practitioner. Additional valuable team members may include a certified athletic trainer, an exercise physiologist, and the athlete's coach, parents and other family members. The first aim of treatment for any Triad component is to increase energy availability by increasing energy intake and/or reducing exercise energy expenditure. Nutrition counseling and monitoring are sufficient interventions for many athletes, but eating disorders warrant psychotherapy. Athletes with eating disorders should be required to meet established criteria to continue exercising, and their training and competition may need to be modified. No pharmacological agent adequately restores bone loss or corrects metabolic abnormalities that impair health and performance in athletes with functional hypothalamic amenorrhea. PMID- 17909418 TI - Important insight from the 2003 Singapore half-marathon. PMID- 17909420 TI - Prescribing, quantifying, and monitoring exercise intensity during interval training. PMID- 17909422 TI - Does antioxidant supplementation prevent favorable adaptations to exercise training? PMID- 17909425 TI - Political competency: part of the nurse leader role. PMID- 17909426 TI - Advocacy: central to the role of nurses: an interview with Colleen Scanlon by Kathleen D. Sanford. PMID- 17909427 TI - The value of the political action committee: dollars and influence for nurse leaders. AB - Nurse leaders play an important role in the shaping of healthcare policy and the care delivery models of the future. It is vital for nurse leaders to understand the role of money in politics and how donations to a candidate's campaigns can be beneficial in getting one's concerns addressed by Congress. The political action committee (PAC) is a major vehicle for campaign contributions for candidates from any party. This article will detail the following related to PACs: history and background information, government regulations, and why PACs are an important vehicle for nurse leaders to gain access and influence in Congress. PMID- 17909428 TI - Acuity systems dialogue and patient classification system essentials. AB - Obtaining resources for quality patient care is a major responsibility of nurse leaders and requires accurate information in the political world of budgeting. Patient classification systems (PCS) assist nurse managers in controlling cost and improving patient care while appropriately using financial resources. This paper communicates acuity systems development, background, flaws, and components while discussing a few tools currently available. It also disseminates the development of a new acuity tool, the Patient Classification System. The PCS tool, developed in a small rural hospital, uses 5 broad concepts: (1) medications, (2) complicated procedures, (3) education, (4) psychosocial issues, and (5) complicated intravenous medications. These concepts embrace a 4-tiered scale that differentiates significant patient characteristics and assists in staffing measures for equality in patient staffing and improving quality of care and performance. Data obtained through use of the PCS can be used by nurse leaders to effectively and objectively lobby for appropriate patient care resources. Two questionnaires distributed to registered nurses on a medical surgical unit evaluated the nurses' opinion of the 5 concepts and the importance for establishing patient acuity for in-patient care. Interrater reliability among nurses was 87% with the author's acuity tool. PMID- 17909429 TI - Negotiating for clinical IT dollars: lessons learned. AB - Politically savvy nurse leaders develop negotiating and fact-based presentation skills. Referencing point-of-care medication administration systems and elimination of healthcare acquired infections as sample initiatives, this article shares concepts for successful presentations to approval boards that select and approve IT projects. Demonstrating diligence in developing realistic cost models and metrics for clinical patient quality improvement projects enables the nurse leader to gain the momentum needed to keep the clinical project positioned as a priority for budget approval. PMID- 17909430 TI - Health services research methods: tools for nurse leaders. AB - A political competency for leaders is to effectively articulate the evidence behind management best practices. Evidence-based practice requires special skills from the nurse leader, many of which are found in health services research (HSR) methods. This review presents approaches associated with HSR, which can be used by nurse managers for the benefit of their units. HSR methods reviewed are cost analyses, small area analysis, geographic information systems, use of existing databases, quality of care measures, and risk adjustment. This review examines the kind of evidence various HSR methods provide, as well as examples of their use and resources needed to apply them. PMID- 17909431 TI - The development of caring in the perioperative culture: nurse leaders' perspective on the struggle to retain sight of the patient. AB - This article focuses on Swedish nurse leaders and is aimed at achieving a more complete and differentiated understanding of what constitutes caring in the perioperative culture as well as their knowledge and responsibility for the development of caring. Interviews with open-ended questions were conducted with 10 nurse leaders, in which they described their experiences of developing perioperative caring. The interpretation process was based on Gadamer's philosophy of hermeneutics. The findings indicate that developing a perioperative caring culture is a struggle to retain sight of the patient, a process that includes the following 6 phases: (1) when the nurse leaders understood perioperative caring as a process, the nurse's and patient's shared world became obvious to them; (2) safeguarding the patient's position as a unique human being; (3) safeguarding the nurse's welfare by creating a compassionate atmosphere; (4) promoting an idea means never giving up; (5) attaching importance to being trustworthy; and (6) being involved in a dynamic interaction, comprising communion and reciprocity. The most important goal of nursing leadership is to safeguard the welfare of the suffering patient and the relationship between the nurse leader and nursing staff, based on the motive of caritas derived from the idea of humanistic caring. PMID- 17909432 TI - Nurse manager support: what is it? Structures and practices that promote it. AB - Professional nursing organizations identify nurse manager (NM) support of staff nurses as an essential component of a productive, healthy work environment. Role behaviors that constitute this support must be identified by staff nurses. In this mixed-method study, supportive role behaviors were identified by 2382 staff nurses who completed the investigator-developed Nurse Manager Support Scale. In addition, semistructured interviews were conducted with 446 staff nurses, managers, and physicians from 101 clinical units in 8 Magnet hospitals in which staff nurses had previously confirmed excellent nurse manager support. Through individual and focus group interviews with NM and chief nurse executives in the 8 participating hospitals, the organizational structures and practices that enabled NM to be supportive to staff were determined. The 9 most supportive role behaviors cited by interviewees were as follows: is approachable and safe, cares, "walks the talk," motivates development of self-confidence, gives genuine feedback, provides adequate and competent staffing, "watches our back," promotes group cohesion and teamwork, and resolves conflicts constructively. Supporting structures and programs identified by managers and leaders include the following: "support from the top," peer group support, educational programs and training sessions, a "lived" culture, secretarial or administrative assistant support, private office space, and computer classes and seminars. PMID- 17909434 TI - The politics of information technology. AB - Information technology implementation is a political process. In the increasingly cost-controlled, high-tech healthcare environment, a successful nursing system implementation demands a nurse leader with both political savvy and technological competency. PMID- 17909433 TI - Interdisciplinary shared decision-making: taking shared governance to the next level. AB - Healthcare organizations have been challenged to make high-quality, safe, and reliable care more affordable, portable, transparent, and efficient. Skillful leadership and contributions from all members of the organization are needed to accomplish these changes. This article describes how one organization, driven by a vision for finding better ways, embarked on a journey to design, implement, and refine their model for interdisciplinary shared decision-making (SDM). The model was based on the principles of accountability, equity, ownership, and partnership, and designed to support staff members in right decision-making at the point of service. The success of SDM depends on leaders who believe in the process and demonstrate their commitment by holding front-line managers accountable for allocating resources and coaching staff. Point-of-service staff, in turn, must accept ownership and accountability for the content of their work and be willing to develop their SDM knowledge, comfort, and skills over time. Preliminary outcomes indicate that these new processes have had a positive effect in helping the organization find better ways to innovate and improve patient care. PMID- 17909435 TI - Regulatory politics from the corporate board to the nursing board: five political issues facing regulators. PMID- 17909437 TI - Sigmoid sinus thrombosis as a complication of otitis media. PMID- 17909438 TI - Lateral sinus thrombosis in chronic otitis media. PMID- 17909436 TI - The contribution of GJB2 (Connexin 26) 35delG to age-related hearing impairment and noise-induced hearing loss. AB - HYPOTHESIS: The common GJB2 (Connexin 26) 35delG mutation might contribute to the development of age-related hearing impairment (ARHI) and noise-induced hearing loss (NIHL). BACKGROUND: GJB2, a gene encoding a gap junction protein expressed in the inner ear, has been suggested to be involved in the potassium recycling pathway in the cochlea. GJB2 mutations account for a large number of individuals with nonsyndromic recessive hearing loss, with 35delG being the most frequent mutation in populations of European origin. Other genes involved in potassium homeostasis have been suggested to be associated with ARHI and NIHL, and distortion product otoacoustic emission distortions indicative of hearing loss alterations have been found in 35delG carriers. METHOD: We genotyped 35delG in two distinct sample sets: an ARHI sample set, composed of 2,311 Caucasian samples from nine different centers originating from seven different countries with an age range between 53 and 67 years, and an NIHL sample set consisting of 702 samples from the two extremes of a noise-exposed Polish sample. RESULTS: After statistical analysis, we were unable to detect an association between 35delG and ARHI, nor between 35delG and NIHL. CONCLUSION: Our findings indicate that there is no increased susceptibility in 35delG carriers for the development of ARHI or NIHL. PMID- 17909439 TI - Use of Bioglue in translabyrinthine vestibular schwannoma surgery. PMID- 17909440 TI - Comment on Ceylan et al., "Impact of Jacobson's (tympanic) nerve sectioning on middle ear functions". PMID- 17909441 TI - Otosclerosis in the incus: fact or fantasy? PMID- 17909442 TI - Surgical management of lesions of the internal carotid artery using a modified Fisch Type A infratemporal approach. PMID- 17909443 TI - Letter to editor re: anterior canal positioning vertigo treatment method. PMID- 17909445 TI - Migration of Cymetra after vocal fold injection for laryngeal paralysis. AB - Injection laryngoplasty is a common operative technique used in the management of unilateral vocal fold paralysis. Cymetra is a micronized particulate injectable form of acellular human dermis that is commonly used for vocal cord medialization procedures. We report migration of an intracordal bolus of Cymetra into the medial wall of the pyriform sinus. Histopathology from endoscopic resection of the migrated Cymetra demonstrated a localized foreign body reaction with characteristic giant cells. Specific anatomic considerations through cadaveric laryngeal dissection are presented to demonstrate the likely pathway of intralaryngeal bolus migration. PMID- 17909446 TI - Bacteriologic comparison of tonsil core in recurrent tonsillitis and tonsillar hypertrophy. AB - OBJECTIVES: Although many bacteriology studies on tonsillar diseases have been completed, all have been confined to children and were characterized by a paucity of cases. The purpose of this study was to analyze the underlying bacterial pathogens in tonsillar disease. METHODS: A retrospective study was performed on 824 patients who underwent elective tonsillectomy with or without adenoidectomy. We analyzed the differences between the bacterial pathogens in recurrent tonsillitis and tonsillar hypertrophy with regard to age, season, and antibiotic sensitivity. RESULTS: Among 824 cases, 966 bacterial strains from the tonsil core were isolated. In recurrent tonsillitis, Staphylococcus aureus was the most common pathogen (30.3%), followed by Haemophilus influenzae (15.5%) and group A beta-hemolytic Streptococcus (Streptococcus pyogenes, 14.4%). In patients over 14 years of age, quite differently from other age groups, Klebsiella pneumoniae was isolated at a significantly higher percentage. In tonsillar hypertrophy, H. influenzae was isolated most commonly (31.4%) regardless of age, followed by S. pyogenes (24.2%), S. aureus (22.9%), and Streptococcus pneumoniae (12.6%). Furthermore, mixed infection was common because of its high resistance to penicillin. In both groups, S. pneumoniae was more common in younger patients, whereas K. pneumoniae was relatively common in adults. We found no differences in the detection rate by season; however, H. influenzae was frequently isolated in the tonsillar hypertrophy group regardless of seasonal variations. We also found no difference in the antibiotic sensitivity between the two groups; however, strains resistant to penicillin were relatively prevalent and showed a high sensitivity to third-generation cephalosporin. CONCLUSIONS: We observed some differences in the types of bacteria in the tonsillar core between the recurrent tonsillitis and tonsillar hypertrophy groups. Our study indicates that essential bacteria have been changing and, thus, we need to change our choice of antibiotics. PMID- 17909447 TI - Scala tympani cochleostomy II: topography and histology. AB - OBJECTIVE: To assess intracochlear trauma using two different round window related cochleostomy techniques in human temporal bones. METHODS: Twenty-eight human temporal bones were included in this study. In 21 specimens, cochleostomies were initiated inferior to the round window (RW) annulus. In seven bones, cochleostomies were drilled anterior-inferior to the RW annulus. Limited cochlear implant electrode insertions were performed in 19 bones. In each specimen, promontory anatomy and cochleostomy drilling were photographically documented. Basal cochlear damage was assessed histologically and electrode insertion properties were documented in implanted bones. RESULTS: All implanted specimens showed clear scala tympani electrode placements regardless of cochleostomy technique. All 21 inferior cochleostomies were atraumatic. Anterior-inferior cochleostomies resulted in various degrees of intracochlear trauma in all seven bones. CONCLUSION: For atraumatic opening of the scala tympani using a cochleostomy approach, initiation of drilling should proceed from inferior to the round window annulus, with gradual progression toward the undersurface of the lumen. While cochleostomies initiated anterior-inferior to the round window annulus resulted in scala tympani opening, many of these bones displayed varying degrees of intracochlear trauma that may result in hearing loss. When intracochlear drilling is avoided, the anterior bony margin of the cochleostomy remains a significant intracochlear impediment to in-line electrode insertion. PMID- 17909448 TI - Treatment of Meniere's disease by low-dosage intratympanic gentamicin application: effect on otolith function. AB - OBJECTIVES: The intratympanic application of a low dosage of gentamicin is increasingly favored as treatment for Meniere's disease. While posttreatment observations have confirmed a long-term success of the therapy of vertigo attacks, clear differences in the posttreatment recovery interval can be observed. In addition to differences in central-vestibular compensation, the degree of peripheral vestibular damage, i.e., to the saccule, utricle, and semicircular canal ampullae, varies among patients. This study provides comprehensive pre- and posttreatment results from unilateral functional tests of the individual vestibular receptors and of the cochlea in patients with Meniere's disease. STUDY DESIGN: Prospective clinical study. METHODS: Nineteen patients with unilateral Meniere's disease were treated by intratympanic application of gentamicin by injection of 0.3 mL (12 mg) through the tympanic membrane under local anesthesia. Tests were performed immediately previous to treatment and subsequently in the periods 4 to 8 weeks and 12 to 16 weeks after treatment. Unilateral saccular function was tested by means of acoustic-click, vestibular evoked myogenic potentials (VEMP), and unilateral utricular function by subjective visual vertical (SVV) during unilateral centrifugation. Bithermal caloric testing was performed to assess unilateral semicircular canal function. RESULTS: Prior to gentamicin treatment, the caloric response from the diseased ear was normal in 3 patients, below normal in 14 patients, and in 2 cases almost completely absent. VEMP responses could be recorded bilaterally in 13 patients; while in 6, no VEMPs could be measured from the diseased ear. Utricular function measured by SVV estimation was found to be normal in 11 patients and marginally abnormal in 2 patients. In six cases, the SVV was clearly underestimated during centrifugation of the diseased side. The posttreatment findings demonstrate that VEMPs were absent in all treated patients, and the caloric response was abnormally low in all but one case. In contrast, only 12 of 19 patients produced abnormal SVV responses. CONCLUSION: The results demonstrate that incremental, intratympanic application of gentamicin effectively eliminates semicircular canal and saccular function. In contrast, utricular function appears to be maintained in 30 to 40% of cases. PMID- 17909449 TI - Cervical discitis and epidural abscess after tonsillectomy. AB - Approximately 6 weeks after an uncomplicated tonsillectomy for chronic tonsillitis, a 37-year-old woman presented to our emergency department with complaints of odynophagia and cervical pain persistent since surgery. Computed tomographic and magnetic resonance imaging revealed cervical spinal osteomyelitis with epidural abscess at C2 to 3. The patient underwent treatment with intravenous antibiotics, operative debridement, and cervical spinal stabilization. She recovered with no neurologic deficit. CONCLUSIONS: Significant infectious complications of tonsillectomy are uncommon, and cervical spinal osteomyelitis and epidural abscess are exceptionally rare occurrences. In the presence of prolonged pain and dysphagia, imaging can be considered to evaluate for such sequelae. PMID- 17909450 TI - Effect of N-acetylcysteine on carboplatin-induced ototoxicity and nitric oxide levels in a rat model. AB - OBJECTIVE: The aim of the present study is to investigate the effect of N acetylcysteine (NAC) given 30 minutes before carboplatin administration on carboplatin-induced ototoxicity and nitric oxide (NO) levels in a rat model. STUDY DESIGN: Animal study. METHODS: Eighteen Sprague-Dawley rats were divided into three groups that each contained six animals. Intraperitoneal injection of physiologic saline was performed in group 1 twice with an interval of 30 minutes. Group 2 was treated with a single bolus administration of carboplatin at a dose of 256 mg/kg 30 minutes after the intraperitoneal injection of physiologic saline. Group 3 was treated with a single bolus administration of carboplatin at a dose of 256 mg/kg 30 minutes after the intraperitoneal injection of NAC at a dose of 400 mg/kg. Pretreatment and posttreatment distortion product otoacoustic emissions (DPOAE) were performed in rats from all groups. Then, the animals were sacrificed on the fourth day, and cochlear tissue NO and glutathione peroxidase (GSH-Px) levels were measured. RESULTS: The comparison of pre- and posttreatment DPOAE responses did not demonstrate any significant changes for groups 1 and 3. Results of group 2 showed a decrease of the DPOAE amplitude. Cochlear NO levels were significantly higher in rats treated with carboplatin than in controls and in those treated with carboplatin plus NAC (P < .05). Cochlear GSH-Px levels were higher in rats treated with carboplatin plus NAC than in those treated with carboplatin, but the difference did not reach statistical significance (P = .079). CONCLUSIONS: The present study showed that carboplatin at higher doses induced hearing loss and increased NO levels in the cochlea of rats. NAC appears to have a protective effect against carboplatin-induced ototoxicity, which may be related to its inhibitory effect on NO production. PMID- 17909451 TI - Chronic ear surgery in patients with syndromes and multiple congenital malformations. AB - OBJECTIVES/HYPOTHESIS: The propensity for the development of chronic ear disease in patients with certain congenital syndromes is well described. Little is known about the efficacy of surgery in the management of such patients. This paper will review an institutional experience with the surgical management of chronic ear disease in patients with congenital syndromes or multiple major malformations associated with high rates of ear disease. STUDY DESIGN: Retrospective chart review. METHODS: Charts were reviewed to identify patients diagnosed with either a congenital syndrome or multiple major malformations with a known association with the development of chronic ear disease who underwent ear surgery for chronic ear disease (excluding tympanostomy tube placement). Syndromes encountered, surgeries performed, operative outcomes, complications, and the efficacy of ossicular chain reconstruction is reported. RESULTS: Forty-three patients with 14 different syndromes or malformations were identified. These patients underwent 66 surgical procedures on 56 ears. Seventy-nine percent of patients had undergone an ear procedure prior to presentation. Disease eradication was achieved in 64% of ears with a single procedure, and 89% of ears were controlled with two surgeries or less. Thirty-two percent of surgeries involved a canal wall down procedure, a rate similar to that seen for all patients in our practice over the past decade. When used, ossicular chain reconstruction significantly reduced the air-bone gap, resulting in hearing improvement. Results for patients with Down syndrome, Turner syndrome, and conotruncal cardiac abnormalities are discussed. Only minor complications were encountered. CONCLUSIONS: Syndromic patients and those with a major congenital malformation may present with significant chronic ear disease. Appropriate surgical management can yield successful eradication of disease with low complication rates. PMID- 17909452 TI - Do the cardiovascular disease risks and benefits of oral versus transdermal estrogen therapy differ between perimenopausal and postmenopausal women? PMID- 17909453 TI - Hexosamine biosynthesis and protein O-glycosylation: the first line of defense against stress, ischemia, and trauma. AB - An early and rapid response to severe injury or trauma is the development of hyperglycemia, which has long been thought to be an essential survival response by providing fuel for vital organ systems and facilitating mobilization of interstitial fluid reserves by increasing osmolarity. However, glucose can also be metabolized via the hexosamine biosynthesis pathway (HBP), leading to the synthesis of uridine diphosphate N-acetyl-glucosamine(UDP-GlcNAc). UDP-GlcNAc is a substrate for the addition, via an O-linkage, of a single N-acetylglucosamine to serine or threonine residues of nuclear and cytoplasmic proteins (O glycosylation, O-GlcNAc). There is increasing appreciation that protein O glycosylation is a highly dynamic posttranslational modification that plays a key role in signal transduction pathways. Sustained increases in O-GlocNAc have been implicated in the development of diabetes and diabetic complications; however, recent studies have demonstrated that stress leads to a transient increase in O GlcNAc levels that is associated with increased tolerance to stress. Indeed, activation of pathways leading to O-GlcNAc formation improves cell survival after I/R injury, whereas inhibition of O-GlcNAc formation decreases cell survival. In addition, in rodent models of trauma-hemorrhage, increasing O-GlcNAc levels during resuscitation improves cardiac function and organ perfusion and attenuates the inflammatory response. At the cellular level, increasing O-GlcNAc levels attenuates nuclear factor-kappaB activation. It is noteworthy that other metabolic-based treatments for severe injury such as glucose-insulin-potassium and glutamine also lead to increased HBP flux and O-GlcNAc levels. The goal of this review is to summarize our current understanding of the role of the HBP and O-GlcNAc on the regulation of cell function and survival and to present evidence to support the notion that activation of these pathways represents a novel treatment strategy for severe injury and trauma. PMID- 17909455 TI - Systemic IL-17 after severe injuries. AB - IL-17 is a cytokine produced by a newly identified T-cell subpopulation (THl7/THIL-17). It is a central mediator in inflammatory processes that connects T-cell stimulation with neutrophil mobilization. The role of IL-17 in the immune dysfunction after polytrauma is still not clarified. In a retrospective study, the systemic concentration of IL-17 and IL-6 of 71 polytraumatized patients were analyzed daily by enzyme-linked immunosorbent assay. The patients' collective consist of 55 men and 16 women (43 +/- 16 years; injury severity score, 33 +/- 13). In only 6% of the patients, an increase in systemic IL-17 was detected. In most patients (94%), no systemic IL-17 was detectable or the IL-17 concentrations in plasma were in the range of the healthy donor group. To identify a possible role of systemic IL-17 in the posttraumatic phase, the patients were divided into two groups. Group A (47 men, 15 women) consists of patients with IL-17 concentrations in the range of normal healthy donors. Group B (8 men, 1 woman) consists of patients with elevated (>45 pg ml(-1) on at least 3 consecutive days) systemic IL-17 concentrations. Three patients in group B showed highly increased systemic IL-17 concentrations (median, >200 pg mL(-1)). These patients were male and showed all blunt chest and abdominal trauma with lung contusion and pneumohemothorax. However, there was no conformity in other injury patterns, injury severity score, age, outcome, intensive care period, or clinical complications. After a period of 4 years, we were able to obtain a new blood sample from one patient with high IL-17 level. The systemic IL-17 value of this former patient was now less than the detection limit. However, stimulation of peripheral blood mononuclear cells from thlise patient revealed elevated numbers of cells with the capacity to produce IL-17 as determined by enzyme-linked immuno spot assay and flow cytometry compared with peripheral blood mononuclear cells obtained from current polytrauma patients and healthy donors. In conclusion, IL 17 is not suitable as a pathophysiological or predictive marker after polytrauma. Whether highly increased systemic IL-17 concentrations detected in single patients are due to individually increased numbers of TH17 cells as we have demonstrated with one rerecruited patient has to be further analyzed. PMID- 17909454 TI - Distribution of NOS isoforms in a porcine endotoxin shock model. AB - Sepsis is a major cause of morbidity and mortality. NO, an endogenous vasodilator, has been associated with the hypotension, catecholamine hyporesponsiveness, and myocardial depression of septic shock. Although iNOS is thought to be responsible for the hypotension and loss of vascular tone occurring several hours after endotoxin administration, little is known on the effects of constitutive eNOS on LPS-induced organ dysfunction. This study assessed the distribution of eNOS and iNOS in various vascular beds in conscious pigs challenged with LPS. Cardiac and regional hemodynamic parameters were recorded over 8 h in the presence and absence of aminoguanidine, a rather selective inhibitor of iNOS activity, and N-methyl-L-arginine, a nonspecific NOS inhibitor. Our data show that LPS-induced cardiac depression was associated with coronary, renal, and mesenteric vasoconstrictions and a hepatic vasodilatation. LPS also induced increases in eNOS in the heart and lungs, whereas iNOS was mostly detected in the liver. Nitrotyrosine formation was mainly detected in the lungs, with traces in the kidney, liver, and gut. Accordingly, our results suggest that the early decrease in blood pressure and cardiac depression are likely due to activated eNOS, whereas both isoforms are involved in the hepatic vasodilation. In contrast, carotid, coronary, mesenteric, and renal vasoconstrictions were significant at 5 and/ or 6 h after LPS infusion, suggesting that NO is not the primary mediator, facilitating and/or unmasking the release of vasoconstrictor mediators. Consequently, developing newer tissue- or isoform-specific NOS inhibitors can lead to novel therapeutic agents in septic shock. PMID- 17909457 TI - [Histoseminaries: a multimedia continuing education series]. PMID- 17909456 TI - Effects of prednisolone on the systemic release of mediators of cell-mediated cytotoxicity during human endotoxemia. AB - Corticosteroids are widely used for the suppression of cell-mediated cytoxicity. This process is mediated by natural killer cells and cytotoxic T lymphocytes, and their activation can be monitored by levels of the chemokines CXCL9 and CXCL10, the degranulation product granzymes A and B, and by levels of secretory phospholipase A2. The current study aimed to determine the effects of increasing doses of prednisolone on the release of these mediators in healthy humans exposed to LPS. Therefore, 32 healthy men received prednisolone orally at doses of 0, 3, 10, or 30 mg (n = 8 per group) at 2 h before intravenous injection of Escherichia coil LPS (4 ng/kg). Prednisolone dose-dependently attenuated the LPS-induced rises in the plasma concentrations of the chemokines CXCL9 and CXCL10, as well as of granzymes A and B levels. CXCL10 and granzyme B release were most sensitive to prednisolone, with a significant inhibition already achieved at the lowest prednisolone dose (3 mg). The levels of secretory phospholipase A2 were increased after LPS administration but were not significantly affected by prednisolone. This study demonstrates that prednisolone differentially inhibits the systemic release of mediators involved in cell-mediated cytotoxicity in humans in vivo. PMID- 17909458 TI - [Obituary: Andre Mazabraud]. PMID- 17909460 TI - [Diagnostic progress in liver tumor pathology]. PMID- 17909459 TI - [Mohs Micrographic Surgery for the management of basal cell carcinoma: experience at the Nimes Univeristy Hospital]. AB - AIMS: The goal of this work was to assess the validity of Mohs Micrographic Surgery (MMS) for basal cell carcinoma (BCC) in a routine clinical setting. MATERIAL AND METHODS: Our adaptation of the technique described by Mohs and coll allows intraoperative histological examination of all surgical edges of the resection. Sixteen men and 4 women were selected. RESULTS: Average operative time was 2 hours 30 minutes. No false results were noted. The cosmetetic and functional outcomes were good. CONCLUSION: MMS is a safe and reproducible surgical technique made possible by solid team work. It is adapted for the treatment of BCC with a high risk of recurrence. The cosmetetic and functional results are quite satisfactory. The recurrence rate at 5 years is 10 times less than with other methods of treatment. The additional time required for this surgery be put in balance with the number of tumors for which a second intervention would have been necessary if conventional surgery had been used. All procedures were performed under local anesthesia, and none or the patients required a second intervention. PMID- 17909463 TI - [Case 1: Hepatocellular adenoma and Case 2: Inflammatory/telangiectasic adenoma]. PMID- 17909464 TI - [Case 3: High grade dysplastic nodule with focal hepatocellular carcinoma]. PMID- 17909465 TI - [Case 4: Fibrolamellar hepatocellular carcinoma]. PMID- 17909466 TI - [Case 5: Intrahepatic cholangiocarcinoma]. PMID- 17909467 TI - [Case 6: Combined hepatocellular and cholangiocarcinoma]. PMID- 17909468 TI - [Case 7: Intrabiliary metastasis of colonic carcinoma]. PMID- 17909469 TI - [Case 8: Liver metastasis of pancreatic acinar cell carcinoma]. PMID- 17909470 TI - [Answers]. PMID- 17909471 TI - [Answers]. PMID- 17909472 TI - [Primary cutaneous plasmacytoma]. AB - Primary cutaneous plasmacytoma (PCP) is a rare cutaneous B cell lymphoma. We report a case of PCP in a 64 year old woman presenting with a nodular lesion of the left cheek. Histologically, the lesion was composed predominately of variably maturated plasma cells with monotypic expression of lambda chain. Extracutaneous localizations of the disease had been excluded. The prognosis of PCP is better than that of the metastatic cutaneous lesion of myeloma. The main prognosis factors are the size tumor and clinical presentation (solitary, versus multiple lesions). Solitary lesions of the PCP are treated by surgical excision and sometimes local radiotherapy. PMID- 17909473 TI - [Plexiform schwannoma of the delto-pectoral area]. AB - Plexiform schwannoma is a form of schwannoma which usually involves cutaneous tissues. It cannot be easily differentiated from malignant tumors, especially deep or cellular lesions. We report a deep plexiform schwannoma which we place among the various benign or malignant nerve sheath tumors, which may or may not develop within the context of genetic disease. Finally, the differential diagnoses are discussed. Recognition of the lesion is necessary for appropriate treatment. PMID- 17909474 TI - [TSC2/PKD1 contiguous gene syndrome. Report of two cases]. AB - The two major genes responsible for autosomal dominant polycystic kidney disease and complex tuberous sclerosis are located on chromosome 16 at position 16p13.3, separated by only a few nucleotides. A simultaneous loss of both genes has been termed "the TSC2/PKD1 contiguous gene syndrome". It has been described essentially in young children. We report 2 new cases in French adults, in whom the diagnosis has been made fortuitously on the macroscopic and microscopic examination of the nephrectomy specimen. This diagnosis should be considered for the association of a polycystic kidney disease and numerous angiomyolipomas. It is necessary to set up a specific follow-up of both diseases. PMID- 17909475 TI - [Pneumonia caused by Cunninghamella bertholletiae in a patient with acute lymphoblastic leukemia]. AB - Pulmonary mucormycosis occur in immunosuppressed patients with a neutropenic treatment or a malignant neoplasia. Although the histological study alone cannot enable species identification, the pathologist usually makes the diagnosis of mucormycosis and can eliminate other filamentous mycoses such as aspergillosis or scedosporiosis. Pulmonary infection caused by Cunninghamella bertholletiae is a very rare lung mucormycosis, particularly in young patients. The histological features of this filamentous mycosis are unusual making the diagnosis of mucormycosis difficult. Mycological study is then crucial for the diagnosis. We report a case of a pulmonary infection caused by Cunninghamella bertholletiae occurring in a young girl with acute lymphoblastic leukemia. PMID- 17909476 TI - [A unusual uterine tumor]. PMID- 17909477 TI - [A congenital tumor of the rhinopharynx]. PMID- 17909478 TI - [An exceptional cause of inguinal hernia]. PMID- 17909479 TI - [A myxoid tumor of leg]. PMID- 17909480 TI - [A rare tumor of soft tissues of the foot]. PMID- 17909481 TI - [A misleading lesion of the penis]. PMID- 17909482 TI - [3D Histology: a protocol of micrographic surgery well fitted for French dermatologists and pathologists]. AB - We present 2 techniques of micrographic surgery (3D-histology) useful to control lateral and deep limits of cutaneous tumors. We have adapted the protocol created in Germany, at the University of Tuebingen, for French pathologists and dermatologists. PMID- 17909483 TI - Synthesis of novel nitro-substituted triaryl pyrazole derivatives as potential estrogen receptor ligands. AB - Novel tetrasubstituted pyrazole derivatives bearing a nitro substituent on their A-phenol ring were synthesized and their binding affinity towards the estrogen receptor (ER) subtypes ERalpha and ERbeta was determined. Among compounds tested, the 2-nitrophenol derivative 5c was found to bind satisfactorily to both estrogen receptor subtypes (RBAalpha=5.17 and RBAbeta=3.27). In general, the introduction of a nitro group into the A ring of these compounds was found to benefit their ERbeta binding abilities. PMID- 17909485 TI - Flavonoids and strigolactones in root exudates as signals in symbiotic and pathogenic plant-fungus interactions. AB - Secondary plant compounds are important signals in several symbiotic and pathogenic plant-microbe interactions. The present review is limited to two groups of secondary plant compounds, flavonoids and strigolactones, which have been reported in root exudates. Data on flavonoids as signaling compounds are available from several symbiotic and pathogenic plant-microbe interactions, whereas only recently initial data on the role of strigolactones as plant signals in the arbuscular mycorrhizal symbiosis have been reported. Data from other plant microbe interactions and strigolactones are not available yet. In the present article we are focusing on flavonoids in plant-fungal interactions such as the arbuscular mycorrhizal (AM) association and the signaling between different Fusarium species and plants. Moreover the role of strigolactones in the AM association is discussed and new data on the effect of strigolactones on fungi, apart from arbuscular mycorrhizal fungi (AMF), are provided. PMID- 17909484 TI - (-)-Catechin in cocoa and chocolate: occurrence and analysis of an atypical flavan-3-ol enantiomer. AB - Cocoa contains high levels of different flavonoids. In the present study, the enantioseparation of catechin and epicatechin in cocoa and cocoa products by chiral capillary electrophoresis (CCE) was performed. A baseline separation of the catechin and epicatechin enantiomers was achieved by using 0.1 mol x L(-1) borate buffer (pH 8.5) with 12 mmol x L(-1) (2-hydroxypropyl)-gamma-cyclodextrin as chiral selector, a fused-silica capillary with 50 cm effective length (75 microm I.D.), +18 kV applied voltage, a temperature of 20 degrees C and direct UV detection at 280 nm. To avoid comigration or coelution of other similar substances, the flavan-3-ols were isolated and purified using polyamide-solid phase-extraction and LC-MS analysis. As expected, we found (-)-epicatechin and (+)-catechin in unfermented, dried, unroasted cocoa beans. In contrast, roasted cocoa beans and cocoa products additionally contained the atypical flavan-3-ol ( )-catechin. This is generally formed during the manufacturing process by an epimerization which converts (-)-epicatechin to its epimer (-)-catechin. High temperatures during the cocoa bean roasting process and particularly the alkalization of the cocoa powder are the main factors inducing the epimerization reaction. In addition to the analysis of cocoa and cocoa products, peak ratios were calculated for a better differentiation of the cocoa products. PMID- 17909486 TI - A simplified procedure for indole alkaloid extraction from Catharanthus roseus combined with a semi-synthetic production process for vinblastine. AB - Dried leaves of Catharanthus roseus were extracted with aqueous acidic 0.1 M solution of HCl. Alkaloid-embonate complexes were obtained as precipitates by treating the extract with an alkaline (NaOH) solution of embonic acid (4,4 methylene-bis-3-hydroxynaphtalenecarboxylic acid). The precipitate mainly consisted of catharanthine and vindoline embonates and it was directly used as the starting material for a semi-synthesis of the anti-cancer bisindole alkaloid vinblastine. The coupling reaction involved oxidation of catharanthine in aqueous acidic medium by singlet oxygen ((1)O2), continuously produced in situ by the reaction between H2O2 with NaClO. An excess of NaBH4 was used for the reduction step. Analysis of the reaction mixture indicated a maximum yield of 20% for vinblastine at pH 8.3, based on the initial amount of catharanthine concentration. Direct-injection electrospray ionization mass spectrometry in positive ion mode was used for the identification of vinblastine. The mass spectra of vinblastine were dominated by the corresponding protonated molecular ion [M+H]+ at m/z 811 and the characteristic fragment ions matched with those of the standard compound. PMID- 17909487 TI - Design and synthesis of a coumarin-based acidichromic colorant. AB - This paper describes the fine-tuning of the acidichromic properties of a coumarin containing colorant 1 by incorporation of electron-donating and electron withdrawing substituents on the coumarin moiety. Colorant 1 can undergo two distinct and reversible color changes under both strongly acidic and basic conditions, but not in the presence of gaseous ammonia. The results indicated that the bromo-substituted compound 5b changes from red to yellow when exposed to gaseous ammonia, both in solution and on polycarbonate film, suggesting that an electron-withdrawing group at the 7-position of the coumarin moiety made the enolic hydrogen on 5b more susceptible to deprotonation by a base than in the unsubstituted compound 1. PMID- 17909488 TI - Chlorine anion encapsulation by molecular capsules based on cucurbit[5]uril and decamethylcucurbit[5]uril. AB - Three barrel-shaped artificial molecular capsules 1-3, based on normal cucurbit[5]uril (Q[5]) and decamethylcucurbit[5]uril (Me10Q[5]), were synthesized and structurally characterized by single-crystal X-ray diffraction. Encapsulation of a chlorine anion in the cavity of a Q[5] or Me10Q[5] to form closed a molecular capsule with the coordinated metal ions or coordinated metal ions and water molecules in the crystal structures of these compounds is common. The three complexes [Pr2(C30H30N20O10)Cl3(H2O)13]3+ 3 Cl- x 5 H2O (1), [Sr2(C40H50N20O10)(H2O)4Cl]3+ 3 Cl- x 2 (HCl) 19 H2O (2) and [K(C40H50N20O10)(H2O)Cl] x [Zn(H2O)2Cl2] x [ZnCl4]2- x 2 (H3O)+ x 8 H2O (3) all crystallize as isolated molecular capsules. PMID- 17909489 TI - A novel and efficient synthesis of N,N-dialkylaminoisopropyl- and O alkylisopropyl-2-(1-alkyl-2-oxopropylidene)phosphonohydrazido oximes--potential marine fish toxin analogues. Part 1. AB - A novel and efficient method for the synthesis of N,N-dialkylaminoisopropyl- and O-alkylisopropyl-2-(1-alkyl-2-oxopropylidene)phosphonohydrazido oximes (4) using activated silica as dehydrating agent has been developed. The reaction involves the condensation of substituted diacetyl monoxime and N,N-dialkylaminoisopropyl phosphono hydrazide or O-alkylisopropylphosphono hydrazides and gave the corresponding analogues of a naturally occurring fish toxin in excellent yields under mild conditions. PMID- 17909490 TI - Synthesis of 4-aryl substituted 3,4-dihydropyrimidinones using silica-chloride under solvent free conditions. AB - This paper describes an improved procedure for the efficient and facile synthesis of 4-aryl substituted 3, 4-dihydropyrimidinones under mild reaction conditions with excellent yields using inexpensive silica chloride under solvent free conditions. PMID- 17909492 TI - Structural characterisation by ESI-MS of feruloylated arabino-oligosaccharides synthesised by chemoenzymatic esterification. AB - The chemoenzymatic synthesis of feruloylated arabino-oligosaccharides has been achieved, using a feruloyl esterase type C from Sporotrichum thermophile (StFaeC). The structure of the feruloylated products was confirmed by ESI-MS(n). PMID- 17909491 TI - Synthesis and characterization of a metal complex containing naringin and Cu, and its antioxidant, antimicrobial, antiinflammatory and tumor cell cytotoxicity. AB - The antioxidant activity of flavonoids is believed to increase when they are coordinated with transition metal ions. However, the literature on this subject is contradictory and the outcome seems to largely depend on the experimental conditions. In order to understand the contribution of the metal coordination and the type of interaction between a flavonoid and the metal ion, in this study a new metal complex of Cu (II) with naringin was synthesized and characterized by FT-IR, UV-VIS, mass spectrometry (ESI-MS/MS), elemental analysis and 1H-NMR. The results of these analyses indicate that the complex has a Cu (II) ion coordinated via positions 4 and 5 of the flavonoid. The antioxidant, anti-inflammatory and antimicrobial activities of this complex were studied and compared with the activity of free naringin. The Naringin-Cu (II) complex 1 showed higher antioxidant, anti-inflammatory and tumor cell cytotoxicity activities than free naringin without reducing cell viability. PMID- 17909494 TI - Synthesis and characterization of 5,10,15,20-tetra[3-(3-trifluoromethyl)phenoxy] porphyrin. AB - The newly synthesized 5,10,15,20-tetra[3-(3-trifluoromethyl)phenoxy]porphyrin, TTFMPP, has been characterized using mass spectroscopy, 1H-, 13C- and 19F-NMR, MALDI-TOF mass spectrometry, UV-Vis and fluorescence spectrophotometry, and cyclic voltammetry. The NMR confirmed the structure of the compound and the mass spectrum was in agreement with the proposed molecular formula. The UV-Vis absorption spectrum of TTFMPP shows characteristic spectral patterns similar to those of tetraphenyl porphryin, with a Soret band at 419 nm and four Q bands at 515, 550, 590, and 648 nm. Protonation of the porphyrin with TFA resulted in the expected red shift of the Soret band. Excitation at 419 nm gave an emission at 650 nm. The quantum yield of the porphyrin was determined to be 0.08. Cyclic voltammetry was used to determine the oxidation and reduction potentials of the new porphyrin. Two quasi-reversible one-electron reductions at -1.00 and -1.32 V and a quasi-reversible oxidation at 1.20 V versus the silver/silver chloride reference electrode with tetrabutylammonium tetrafluoroborate as the supporting electrolyte in methylene chloride were observed. PMID- 17909495 TI - Atmospheric pressure microwave assisted heterogeneous catalytic reactions. AB - The purpose of the study was to investigate microwave selective heating phenomena and their impact on heterogeneous chemical reactions. We also present a tool which will help microwave chemists to answer to such questions as "My reaction yields 90% after 7 days at reflux; is it possible to obtain the same yield after a few minutes under microwaves?" and to have an approximation of their reactions when conducted under microwaves with different heterogeneous procedures. This model predicting reaction kinetics and yields under microwave heating is based on the Arrhenius equation, in agreement with experimental data and procedures. PMID- 17909493 TI - Comparison of the biological properties of several marine sponge-derived sesquiterpenoid quinones. AB - Eight naturally occurring marine-sponge derived sesquiterpenoid quinones were evaluated as potential inhibitors of pyruvate phosphate dikinase (PPDK), a C4 plant regulatory enzyme. Of these, the hydroxyquinones ilimaquinone, ethylsmenoquinone and smenoquinone inhibited PPDK activity with IC50's (reported with 95% confidence intervals) of 285.4 (256.4-317.7), 316.2 (279.2-358.1) and 556.0 (505.9-611.0) microM, respectively, as well as being phytotoxic to the C4 plant Digitaria ciliaris. The potential anti-inflammatory activity of these compounds, using bee venom phospholipase A2 (PLA2), was also evaluated. Ethylsmenoquinone, smenospongiarine, smenospongidine and ilimaquinone inhibited PLA2 activity (% inhibition of 73.2 +/- 4.8 at 269 microM, 61.5 +/- 6.1 at 242 microM, 41.0 +/- 0.6 at 224 microM and 36.4 +/- 8.2 at 279 microM, respectively). SAR analyses indicate that a hydroxyquinone functionality and a short, hydroxide/alkoxide side-chain atC-20 is preferred for inhibition of PPDK activity, and that a larger amine side-chain at C-20 is tolerated for PLA2 inhibitory activity. PMID- 17909496 TI - Electrochemical synthesis and structural characterization of a novel mixed valence copper(I)-copper(II) complex: {[bis(ethylenediamine)copper(II)] bis[diiodocuprate(I)]}. AB - A novel, mixed-valent copper(I)-copper(II) complex, {[bis(ethylene diamine)copper(II)] bis[diiodocuprate(I)]} (1), has been prepared by electrochemical dissolution of a sacrificial copper anode in a solution of ethylenediamine (en), I2 and tetraethylammoniumperchlorate (TEAP) as supporting electrolyte in acetonitrile (AcN)and characterized by single-crystal X-ray structure determination. The crystal structure of the complex 1 shows that it consists of a CuI2 polymer formed from I- ligands bridging Cu(I) ions, with a nearly square planar geometry of bivalent Cu(II) atoms chelated by two ethylenediamine ligands. The results also show that direct electrosynthesis of the complex had high current efficiency, purity and electrolysis yield. PMID- 17909497 TI - An efficient and rapid synthetic route to biologically interesting pyranochalcone natural products. AB - An efficient and concise total synthesis of naturally occurring pyranochalcones was achieved from readily available 2,4-dihydroxyacetophenone and 2,4-dihydroxy-6 methoxyacetophenone. The key steps in the synthetic strategy were ethylenediamine diacetate-catalyzed benzopyran formation and aldol reactions. PMID- 17909498 TI - Revised NMR data for incartine: an alkaloid from Galanthus elwesii. AB - Phytochemical studies on Galanthus elwesii resulted in the isolation of five alkaloids: incartine, hordenine, hippeastrine, 8-O-demethylhomolycorine and lycorine. The NMR data given previously for incartine were revised and completed by two-dimensional 1H-1H and 1H-13C chemical shift correlation experiments. In vitro studies on the bioactivity of incartine were carried out. PMID- 17909500 TI - Leufolins A and B, potent butyrylcholinesterase-inhibiting flavonoid glucosides from Leucas urticifolia. AB - New flavonoidal glucosides leufolins A (1) and B (2), have been isolated from the ethyl acetate soluble fraction of the whole plants of Leucas urticifolia. Their structures were elucidated on the basis of extensive analysis of nuclear magnetic resonance (1D and 2D NMR) spectral data. Both of these compounds exhibited significant inhibitory potential against the enzyme butyrylcholinesterase. PMID- 17909499 TI - A facile, mild and efficient one-pot synthesis of 2-substituted indole derivatives catalyzed by Pd(PPh3)2Cl2. AB - 2-Phenylindoles were prepared by heteroannulation of 2-haloaniline derivatives and phenylacetylene under mild conditions in a one-pot reaction catalyzed by Pd(PPh3)2Cl2. PMID- 17909502 TI - Unusual reactivity patterns of 1,3,6,8-tetraazatricyclo-[4.4.1.1(3,8)]-dodecane (TATD) towards some reducing agents: synthesis of TMEDA. AB - N,N,N,N-tetramethylethylenediamine (TMEDA) can be synthesized by simple reduction of 1,3,6,8-tetraazatricyclo-[4.4.1.1.(3,8)]dodecane (TATD), an aminal cage type amine, with formic acid. The aminal can be converted to TMEDA in high yield very easily and in a very short time. We comment on the scope and limitations of the reduction of this aminal and propose a possible reaction mechanism. PMID- 17909501 TI - The anti-apoptosis effects of daidzein in the brain of D-galactose treated mice. AB - The purpose of this study was to explore the neuroprotective effects of daidzein on the apoptotic pathway in the hippocampus and cortex of D-galactose treated mice. For this purpose we have examined the expression of bcl-2 mRNA, bax mRNA and caspase-3 in the hippocampus and cortex of D-galactose-treated mice after fed with 10 or 5 mg/kg of daidzein. The results of in situ hybridization experiments indicate that daidzein could help increase the transcriptions of bcl-2 and decrease the transcriptions of bax in those brain regions of D-galactose-treated mice. Furthermore, immunohistochemical studies showed that daidzein could reduce the expression of caspase-3 in both brain regions. These results suggest that daidzein in soybean can inhibit the D-gal induced apoptosis via Bcl-2/Bax apoptotic pathway and be a potential medical candidate for neurodegeneration therapy. PMID- 17909503 TI - New "green" approaches to the synthesis of pyrazole derivatives. AB - A novel approach to the synthesis of pyrazole derivatives from tosylhydrazones of alpha,beta-unsaturated carbonyl compounds possessing a beta-hydrogen is proposed, exploiting microwave (MW) activation coupled with solvent free reaction conditions. The cycloaddition was studied on three ketones (trans-4-phenyl-3 buten-2-one, beta-ionone and trans-chalcone). The corresponding 3,5-disubstituted 1H-pyrazoles were obtained in high yields and after short reaction times. In order to simplify and point out the green chemistry features of the method, a further improvement was achieved under the same catalytic conditions with a "one pot" synthesis of these heterocyclic compounds, starting directly from their carbonyl precursors via tosylhydrazones generated in situ. For an exhaustive study, the dielectric properties of the solid reaction mixtures were also measured, in order to obtain input data for the numerical simulation of their heating behaviour in the single mode MW cavity which was used for experimental work. In order to supply a valid methodology and tool for measuring the environmental impact, a comparative study between the synthetic route proposed and the classical synthetic route has been carried out. PMID- 17909505 TI - Oral Candidiasis in children and adolescents with cancer. Identification of Candida spp. AB - Oral candidiasis represents a serious problem for children with cancer. The mortality rate of this infection has increased due to fungal septicemia, associated with a primary buccal infection. OBJECTIVE: Identify the Candida spp. in buccal lesions of patients with cancer, establish the predominant species and correlate them to age and sex of the patient, clinical presentation, type of neoplasic disease and cytostatic therapy received. STUDY DESIGN: 62 patients, between 0-16 years, were investigated in a cross sectional study. SAMPLE INCLUSION CRITERIA: Patients with malignant neoplasic disease that were receiving cytostatic treatment and had suspicious lesions of oral candidiasis. Patients with antifungal therapy, active caries, calculus or intraoral appliances were excluded. A clinical evaluation was carried out. The lesion sample was taken and studied by direct exam and culture in CHROMagar-Candida and Sabouraud-Dextrose Agar with chloramphenicol. The identification of the isolated yeast was done by the filamentation test, carbohydrate fermentation and assimilation. RESULTS: Most of the cases (69.35%) were positive to oral candidiasis, C. albicans was the most frequent species found, followed by C. parapsilosis (14.89%), C. tropicalis (12.77%), C. krusei (4.26%), C. glabrata (2.13%) and C. lusitaniae (2.13%). In some cases more than one specie were isolated (9.30%). The most frequent location of the lesion was in the tongue (72.70%). The pseudomembranous candidiasis was the most frequent clinical presentation found (78.71%). There were not significant statistically differences with regard to sex and age of the patient, type of neoplasic disease and cytostatic agent received. CONCLUSION: The results suggest that oral candidiasis is a frequent complication in the pediatric oncological population, being C. albicans the main etiological agent, however, there is an important participation of other Candida species. PMID- 17909506 TI - Prostatic adenocarcinoma with mandibular metastatic lesion: case report. AB - Metastatic lesions of primary tumors, which originate in different parts of the body, comprise almost 1 % of different types of oral cancers. These lesions can affect either bones or soft tissues in the maxillofacial region. Whenever the maxillofacial area is affected, the most common location is in the molar region of the mandible. The clinical presentation of mandibular metastasis follows a clinical pattern characterized by irradiated dental pain in the third molar region. The most frequent sign is parethesia of the area innervated by the mandibular alveolar dental nerve. Differential diagnosis and treatment of these patients can be extremely difficult because there a number of pathologic conditions with similar symptoms and because diagnostic examination can be highly confusing. The aim of this article is to present a case of prostatic adenocarcinoma where the only metastasis was found in the jaw. A literature review will be presented, hoping to contribute to the scarce information regarding this lesion, due to its low frequency and atypical expression of this type of metastasis in terms of etiology, biological behavior and treatment. PMID- 17909504 TI - Comparative evaluation of various total antioxidant capacity assays applied to phenolic compounds with the CUPRAC assay. AB - It would be desirable to establish and standardize methods that can measure the total antioxidant capacity level directly from vegetable extracts containing phenolics. Antioxidant capacity assays may be broadly classified as electron transfer (ET)- and hydrogen atom transfer (HAT)-based assays. The majority of HAT assays are kinetics-based, and involve a competitive reaction scheme in which antioxidant and substrate compete for peroxyl radicals thermally generated through the decomposition of azo compounds. ET-based assays measure the capacity of an antioxidant in the reduction of an oxidant, which changes colour when reduced. ET assays include the ABTS/TEAC, CUPRAC, DPPH, Folin-Ciocalteu and FRAP methods, each using different chromogenic redox reagents with different standard potentials. This review intends to offer a critical evaluation of existing antioxidant assays applied to phenolics, and reports the development by our research group of a simple and low-cost antioxidant capacity assay for dietary polyphenols, vitamins C and E, and human serum antioxidants, utilizing the copper(II)-neocuproine reagent as the chromogenic oxidizing agent, which we haved named the CUPRAC (cupric ion reducing antioxidant capacity) method. This method offers distinct advantages over other ET-based assays, namely the selection of working pH at physiological pH (as opposed to the Folin and FRAP methods, which work at alkaline and acidic pHs, respectively), applicability to both hydrophilic and lipophilic antioxidants (unlike Folin and DPPH), completion of the redox reactions for most common flavonoids (unlike FRAP), selective oxidation of antioxidant compounds without affecting sugars and citric acid commonly contained in foodstuffs and the capability to assay -SH bearing antioxidants (unlike FRAP). Other similar ET-based antioxidant assays that we have developed or modified for phenolics are the Fe(III)- and Ce(IV)-reducing capacity methods. PMID- 17909507 TI - Congenital granular cell tumor (congenital epulis): a lesion of multidisciplinary interest. AB - Congenital granular cell tumor (CGCT), or congenital epulis, is a very uncommon benign soft tissue lesion that usually arises from the alveolar mucosa of neonates and may cause respiratory and feeding problems. We report a case of a 3 day-old female newborn, who presented an intraoral tumor mass which was protruding from her mouth, and compromising feeding. Under general anesthesia, the lesion was completely removed and the patient had an uneventful postoperative course. Clinical features and treatment approaches are presented and discussed, emphasizing the necessity of a multidisciplinary approach in such cases. PMID- 17909508 TI - Secretor status and ABH antigens expression in patients with oral lesions. AB - OBJECTIVES: The aim of this work was to investigate the secretor status of patients with oral pre-cancerous and cancerous lesions and ABH antigens expression in fixed tissue sections of these patients. STUDY DESIGN: To reveal A, B and H antigens in tissue sections of patients with precancerous and cancerous oral lesions (n= 54) we used a modified specific red cell adherence technique (SRCA-test). Normal endothelial cells expressed ABH antigens, the presence of indicator erythrocytes at the lumen of the blood vessels served as a built in positive control. The test results were graded from negative adherence to very strongly positive adherence. Negative adherence was defined as a complete absence of adhered indicator erythrocytes. A strongly positive reaction was defined as a sheet of indicator erythrocytes adhered to the epithelia cells. RESULTS: In 31 of the 54 samples analyzed the test showed slightly positive results on atypical areas, and there was a complete antigen deletion in areas histologically affected by neoplasia. Sixteen samples showed a total absence of ABH antigens in both histologically normal and pathological areas. As a working hypothesis, we propose that areas of SRCA-test negative epithelium are closely related to invasive carcinomas and may be their precursor lesions. Further it is suggested that areas of blood group isoantigen negative epithelium showing atypia, or in some instances near normal histology, may give rise to relatively low grade carcinomas. CONCLUSIONS: Considering these results we suggest the use of this method to monitor probable preneoplastic lesions in risk population, specially in those with no secretor status. PMID- 17909510 TI - Cinnamon products as a possible etiologic factor in orofacial granulomatosis. AB - OBJECTIVES: It has been reported that clinical changes due to hypersensitivity reactions to various foods, preservatives, and oral hygiene products may be consistent with the characteristic signs of orofacial granulomatosis (OFG). The objective of this study was to examine 37 well-documented cases of cinnamon induced contact stomatitis for clinical and histological features consistent with a diagnosis of OFG. STUDY DESIGN: We reviewed the records of the 37 cases to screen them for the clinical and histopathologic features of OFG. RESULTS: Twelve patients showed clinical characteristics of OFG. The most commonly affected site was the gingiva. Focal non-caseating, epitheloid granulomas were observed in four histologic specimens. Multinucleated giant cells were observed in an additional four cases. CONCLUSIONS: Although OFG may have multiple etiologies, it is clear that, in some instances, a hypersensitivity reaction to cinnamon products can elicit lesions consistent with OFG. PMID- 17909509 TI - pH and salivary sodium bicarbonate during the administration protocol for methotrexate in children with leukemia. AB - OBJECTIVE: To analyze the behavior of pH and sodium bicarbonate (NAHCO3) in the saliva of patients with leukemia during the administration protocol for Methotrexate (Mtx). MATERIALS AND METHODS: A controlled clinical essay was carried out on 23 patients between 4 and 18 years of age with high-risk Acute Lymphoblastic Leukemia. Sampling was carried out at To: basal condition; T1: 12 hours after intravenous administration of sodium bicarbonate, before administering Mtx and T2: 3 hours after administering Mtx, the time of maximum concentration. Chiron-Diagnostic 378 equipment was used to determine pH and sodium bicarbonate. The data was interpreted using Analysis of Variance at the 5% significance level. RESULTS: The highest values of sodium bicarbonate were observed at T2, with salivary pH levels remaining within neutrality ranges, diminishing slightly in T1. CONCLUSION. In this study, the dose of sodium bicarbonate considered in the administration protocol of 3 g /m2 Mtx, kept sodium bicarbonate levels in saliva at normal levels and pH neutral. PMID- 17909511 TI - Cornelia de Lange syndrome: a case report. AB - Cornelia de Lange is a genetic syndrome which affects between 1/10.000 and 1/60.000 neonates, but its genetic bases are still not clear. Its principal clinical characteristics are the delay in growth and development, hirsute, structural anomalies in the limbs and distinctive facial characteristic. Dental problems are frequent and include: ogival palate, micrognathia, dental malalignment, delayed teething, microdontic teeth, periodontal disease and dental erosion produced by gastric reflux. Discussed is the case of a 29 year old patient affected by the syndrome in question, which presents the principal clinical characteristics. The patient's general state of health is acceptable, without cardiac or respiratory alterations. The intraoral exploration shows policaries, periodontal disease, persistence of the temporal teeth and ectopic molars. After completing the necessary pre-operatory preparations, the entire odontological treatment was carried out under general aesthesia, due to the patient's total lack of collaboration. PMID- 17909512 TI - Dental treatment for disabled children in the Spanish Public Health System. AB - The Spanish Public Health System is stepping up its efforts to meet all the medical needs of the population. Oral health is of increasing interest for society, especially for parents who are keen for their children to have healthy teeth. Disabled children with both physical and mental disabilities do not always receive the dental care they need. The purpose of this bibliographical review is to evaluate the services provided by the Spanish Public Health System to such children. We have noted marked differences in the types of dental treatment given to these patients in the different Autonomous Communities of Spain. Some, such as Asturias, Navarra and Extremadura, offer specific care for disabled children. Others, such as Ceuta and Melilla, provide more general care. PMID- 17909513 TI - High resolution image in bone biology I. Review of the literature. AB - Bone microstructure has usually been assessed by obtaining samples invasively and analyzing them with conventional histomorphometric methods. Improvements in high resolution image acquisition systems have enabled non-invasive assessment of bone morphology and a more precise 3-D evaluation by means of "virtual biopsies", permitting bone assessment in regeneration or remodeling processes. This review describes the characteristics and limitations of bone assessment using different high-resolution image systems (synchrotron-radiation computed tomography, micro computed tomography, acoustic scanning microscope; micro-magnetic resonance imaging). Morphometric variables that can be obtained from these images are reported and compared with conventional histomorphometric variables. PMID- 17909515 TI - Effect of post space preparation on apical seal: influence of time interval and sealer. AB - OBJECTIVE: To assess the efficacy of two sealants to preserve the apical seal after root canal preparation and cementation of posts at 24 h or 72 h after endodontic treatment. STUDY DESIGN: Sixty human single-root teeth were instrumented and obturated using lateral compaction technique with EndoFill [30] or AH-Plus) [30] and were prepared in one of three ways, leaving a 3 mm gutta percha remnant in all cases: without cast post preparation, with preparation after 24 h or after 72 h. After cementing the posts, the specimens were thermal cycled at 5 and 55 degrees C in water baths, submerged in 2% methylene blue dye for 72 h, embedded in acrylic resin and cut transversally into three 1-mm apical sections. Dye leakage was quantitatively assessed as the percentage leaked area. RESULTS AND CONCLUSION: Comparison of the apical sections showed significant differences in leakage with both sealers among the three preparation groups (p<0.001). No significant differences between sealers were found in any preparation group or in the same sections. PMID- 17909514 TI - Retention of three fissure sealants and a dentin bonding system used as fissure sealant in caries prevention: 12-month follow-up results. AB - BACKGROUND: Bonding agents could be used as fissure sealants. This study compares the retention three fissure sealants (Delton, Delton Plus and Concise) and a filled dentin bonding system (Optibond Solo). METHODS. Fifty-six children aged 7 8 years received fissure sealants either in the four permanent first molars, in the four deciduous second molars, or in all eight of these teeth. Every child received a different sealing material in each quadrant on a random basis. Clinical evaluation at 12 months was performed by a single blind examiner, and the retention was classified as either a success (total retention) or a failure (partial retention or not present). RESULTS: There were no statistically significant differences among the four materials in permanent maxillary molars or deciduous molars. In permanent mandibular molars, Optibond Solo showed a lower percentage of retention (40.9%), significantly different (p=0.002) to that of Delton (89.5%), Delton Plus (87.5%) and Concise (76.5%). CONCLUSION: One bottle dentin bonding system used as a sealant does not improve the retention of conventional fissure sealants. CLINICAL IMPLICATION: Because of the scarcity of studies on the use of dental adhesives as sealants, further studies are warranted for the final support of that conclusion. PMID- 17909516 TI - [The regulation of mast cell migration. Part 1: cell adhesion molecules]. AB - Mast cells take part in multiple pathological processes, in some of which mast cell accumulation is central to pathogenesis. They are also vital factors in many physiological reactions. Therefore it seems to be of great importance to understand the mechanisms underlying mast cell migration into and within tissues. There are many factors that regulate the migration of mast cell progenitors from the blood into tissues and the migration of mature mast cells within tissues, leading to the rapid local accumulation that occurs in diverse pathological conditions. Without any doubt, cell-surface adhesion molecules are central to the migratory process, as they facilitate the binding of cells to extracellular matrix (ECM) proteins. Immature and mature mast cells express different adhesion molecules, especially integrins, that are involved in mast cell adhesion to such ECM proteins as laminin, fibronectin, vitronectin, and collagens. The expression of adhesion molecules alters during mast cell development and maturation. What is more, mast cell adhesion molecule expression and mast cell adhesion to ECM proteins may be regulated by some cytokines. PMID- 17909518 TI - [Housekeeping genes as a reference in quantitative real-time RT-PCR]. AB - The determination of gene expression levels in normal tissue is necessary for the analysis and interpretation of results of gene profiling studies in pathological samples. With the real-time reverse transcription-PCR technique, which enables one to detect the amplification rate during the process, assessment of the amount of gene transcript is fast and accurate. The most important problem in this type of analysis is the variability in the amount of genetic material between samples, caused mostly by changes in the efficiency of mRNA isolation and reverse transcription. Therefore, a reference gene to normalize sample variations is required. Quantification of the mRNA of the target and the reference gene in the sample ensures that the changes in transcript levels will influence both genes equally. To be used as a reference, a gene should show stable, unregulated expression in the analyzed sample type. Housekeeping genes (HKGs) fulfill this criterion and they are used for normalization purposes in most expression studies. However, transcript levels of HKGs can vary between different types of tissue (normal and pathological samples) and under different treatment conditions (drugs and chemicals). The aim of this study was to show the differences and the factors which can influence housekeeping gene expressions. PMID- 17909517 TI - [The regulation of mast cell migration. Part 2: mast cell chemoattractants]. AB - There are many humoral factors that regulate the migration of mast cell progenitors from the blood into tissues and the migration of mature mast cells within tissues, leading to the rapid accumulation that occurs in diverse pathological conditions. First of all, mast cell migration is stimulated by some chemokines, such as RANTES, eotaxin, and IL-8. Moreover, many cytokines induce the migration of mast cells (i.e. SCF, TNF, IL-15). Finally, the migration of mast cells is also stimulated by many other humoral factors, including those involved in inflammatory process, such as C3a, C5a, histamine, PAF, and CRP. Because mast cells play an essential role in diverse physiological and pathological processes, it seems to be of great importance to know the mechanisms underlying the migration of immature and mature mast cells. However, current knowledge about these processes is still insufficient. PMID- 17909519 TI - [Molecular aspects of aminoglycoside nephrotoxicity]. AB - Aminoglycosides are potent bactericidal antibiotics particularly active against aerobic Gram-negative bacteria. This review focuses on the recent concept of a molecular understanding of aminoglycoside-induced nephrotoxicity. As receptor mediated endocytosis plays an important role in the accumulation of aminoglycosides in renal proximal tubules, the biochemical properties of the two main receptors, megalin and cubilin, involved in this process are described. Literature data on megalin and acidic phospholipids as potential targets for preventing aminoglycoside-induced nephrotoxicity are also presented. PMID- 17909520 TI - Walking the line: balancing work and family. PMID- 17909522 TI - The many faces of actin: matching assembly factors with cellular structures. AB - Actin filaments are major components of at least 15 distinct structures in metazoan cells. These filaments assemble from a common pool of actin monomers, but do so at different times and places, and in response to different stimuli. All of these structures require actin-filament assembly factors. To date, many assembly factors have been identified, including Arp2/3 complex, multiple formin isoforms and spire. Now, a major task is to figure out which factors assemble which actin-based structures. Here, we focus on structures at the plasma membrane, including both sheet-like protrusive structures (such as lamellipodia and ruffles) and finger-like protrusions (such as filopodia and microvilli). Insights gained from studies of adherens junctions and the immunological synapse are also considered. PMID- 17909521 TI - Autophagosome formation: core machinery and adaptations. AB - Eukaryotic cells employ autophagy to degrade damaged or obsolete organelles and proteins. Central to this process is the formation of autophagosomes, double membrane vesicles responsible for delivering cytoplasmic material to lysosomes. In the past decade many autophagy-related genes, ATG, have been identified that are required for selective and/or nonselective autophagic functions. In all types of autophagy, a core molecular machinery has a critical role in forming sequestering vesicles, the autophagosome, which is the hallmark morphological feature of this dynamic process. Additional components allow autophagy to adapt to the changing needs of the cell. PMID- 17909523 TI - Micromanaging metastasis. PMID- 17909524 TI - Network news: complete nuclear coverage. PMID- 17909525 TI - Shaping membranes into autophagosomes. PMID- 17909526 TI - Cell fate in the Hand of Plk4. PMID- 17909528 TI - How to dispel myths in the treatment of hepatic encephalopathy? PMID- 17909531 TI - Mild hypothermia for the treatment of acute liver failure--what are we waiting for? PMID- 17909532 TI - Surgical management of rectal prolapse. AB - This article reviews the pathogenesis, clinical presentation and surgical management of rectal prolapse. Full-thickness prolapse of the rectum causes significant discomfort because of the sensation of the prolapse itself, the mucus that it secretes, and because it tends to stretch the anal sphincters and cause incontinence. Treatment of rectal prolapse is primarily surgical. Perineal surgical repairs are well tolerated, but are generally associated with higher recurrence rates. Abdominal repairs involve fixing the rectum to the sacrum by using either mesh or sutures, and tend to have the lowest recurrence rates. If significant preoperative constipation is present, a sigmoid resection can be performed at the time of rectopexy. For many patients, diarrhea and incontinence improve after surgery. Laparoscopic repair of rectal prolapse has similar morbidity and recurrence rates to open surgery, with attendant benefits of reduced length of hospital stay, postoperative pain and wound complications. PMID- 17909533 TI - Therapy insight: Prophylaxis of stress-induced gastrointestinal bleeding in critically ill patients. AB - Stress-induced gastrointestinal bleeding is associated with increased morbidity and mortality in critically ill patients. Within the past few decades, the incidence of stress-induced gastrointestinal bleeding has decreased. Prophylaxis of stress-induced gastrointestinal bleeding, which is aimed at preventing morbidity and mortality, has to be achieved with as few adverse effects as possible. Data indicate that not all critically ill patients need prophylaxis for stress-induced gastrointestinal bleeding. The main risk factors associated with clinically important hemorrhage are mechanical ventilation for >48 h, and coagulopathy (thrombocyte count <50/nl, partial thromboplastin time (PTT) >2 times the upper limit of the normal range, international normalized ratio (INR) >1.5). Ranitidine is more effective than sucralfate for the prevention of clinically important bleeding. Immediate-release omeprazole is as effective as cimetidine, and is more efficient at increasing the intragastric pH. As yet, however, there is no firm evidence that any of the drugs used for prophylaxis of stress-induced gastrointestinal bleeding in critically ill patients decrease mortality or the length of hospital stay. When to stop prophylaxis is decided on clinical grounds rather than on the basis of data from clinical studies. PMID- 17909534 TI - Primer: Applying the new postpolypectomy surveillance guidelines in clinical practice. AB - Colonoscopy is being increasingly used for colorectal cancer screening, which has resulted in a growing cohort of patients who have polyps that require postpolypectomy surveillance. Risk stratification enables postpolypectomy surveillance to be tailored to individual patient needs, and this is one of the fundamental points emphasized by the unified US Multi-Society Task Force on Colorectal Cancer and the American Cancer Society (USMSTF-ACS) guidelines. Most patients do not require intensive surveillance; those patients who have one or two small (<1 cm) adenomas can safely undergo repeat colonoscopy after 5-10 years. Consensus guidelines that merge the recommendations of all societies are more user-friendly than individual guidelines, decrease confusion, and eliminate conflicting recommendations that are a barrier to guideline uptake. Nonetheless, studies have shown that specialists and nonspecialists overutilize colonoscopy for postpolypectomy surveillance, which places a large burden on already strained resources. Barriers to guideline implementation include factors involving the patient, physician, and health-care system. Physician education and widespread implementation of continuous quality improvement programs are required to bridge the gap between the guidelines and their clinical application. PMID- 17909535 TI - A case of mediastinitis following botulinum toxin type A treatment for achalasia. AB - BACKGROUND: A 62-year-old obese, diabetic female underwent endoscopic esophageal injection of botulinum toxin type A (Botox; Allergan, Irvine, CA) for achalasia. The patient presented to her gastroenterologist with chest pain 4 days after the procedure, but no thoracic or gastrointestinal pathologies were identified and the patient was sent home. She presented again the next day with continuing chest pain and newly developed fever. Esophagoduodenoscopy revealed no esophageal leak and a CT scan revealed only mild paraesophageal inflammation. The patient was sent home the same day with antibiotics after a urinalysis suggested presence of a urinary tract infection. The patient presented again 9 days after the procedure with continuing chest pain and fever. INVESTIGATIONS: Chest radiography, electrocardiography, complete blood count, cardiac enzyme levels, basic metabolic panel, urinalysis, Gastrografin (Bracco Diagnostics Inc, Princeton, New Jersey) and barium swallow study, endoscopy, abdominal and chest CT scans, blood culture and wound culture. DIAGNOSIS: Ulceration without perforation of the esophageal mucosa in the area of Botox injections. Unilateral pleural effusion and mediastinitis without abscess formation. MANAGEMENT: Pantoprazole and clonazepam for suspected gastroesophageal reflux and esophageal spasm. Levofloxacin for urinary tract infection. Intravenous antibiotic therapy and acute surgical exploration for possible esophageal rupture. Paraesophageal drain placement, nasogastric tube placement, and parenteral and enteral feeding. PMID- 17909536 TI - rSac3, a novel Sac domain phosphoinositide phosphatase, promotes neurite outgrowth in PC12 cells. AB - Sac domain-containing proteins belong to a newly identified family of phosphoinositide phosphatases (the PIPPase family). Despite well-characterized enzymatic activity, the biological functions of this mammalian Sac domain PIPPase family remain largely unknown. We identified a novel Sac domain-containing protein, rat Sac3 (rSac3), which is widely expressed in various tissues and localized to the endoplasmic reticulum, Golgi complex and recycling endosomes. rSac3 displays PIPPase activity with PI(3)P, PI(4)P and PI(3,5)P(2) as substrates in vitro, and a mutation in the catalytic core of the Sac domain abolishes its enzymatic activity. The expression of rSac3 is upregulated during nerve growth factor (NGF)-stimulated PC12 cell neuronal differentiation, and overexpression of this protein promotes neurite outgrowth in PC12 cells. Conversely, inhibition of rSac3 expression by antisense oligonucleotides reduces neurite outgrowth of NGF stimulated PC12 cells, and the active site mutation of rSac3 eliminates its neurite-outgrowth-promoting activity. These results indicate that rSac3 promotes neurite outgrowth in differentiating neurons through its PIPPase activity, suggesting that Sac domain PIPPase proteins may participate in forward membrane trafficking from the endoplasmic reticulum and Golgi complex to the plasma membrane, and may function as regulators of this crucial process of neuronal cell growth and differentiation. PMID- 17909537 TI - Histone lysine demethylases: emerging roles in development, physiology and disease. AB - The discovery of an increasing number of histone demethylases has highlighted the dynamic nature of the regulation of histone methylation, a key chromatin modification that is involved in eukaryotic genome and gene regulation. A flurry of recent studies has offered glimpses into the specific biological roles of these enzymes and their potential connections to human diseases. These advances have also catalysed a resurgence of interest in epigenetic regulators as potential therapeutic targets. PMID- 17909538 TI - Genetic links between diet and lifespan: shared mechanisms from yeast to humans. AB - Caloric restriction is the only known non-genetic intervention that robustly extends lifespan in mammals. This regimen also attenuates the incidence and progression of many age-dependent pathologies. Understanding the genetic mechanisms that underlie dietary-restriction-induced longevity would therefore have profound implications for future medical treatments aimed at tackling conditions that are associated with the ageing process. Until recently, however, almost nothing was known about these mechanisms in metazoans. Recent advances in our understanding of the genetic bases of energy sensing and lifespan control in yeast, invertebrates and mammals have begun to solve this puzzle. Evidence is mounting that the brain has a crucial role in sensing dietary restriction and promoting longevity in metazoans. PMID- 17909540 TI - Does NT-proBNP testing reduce costs and improve accuracy in the diagnosis of heart failure? PMID- 17909539 TI - Progress and prospects: gene therapy clinical trials (part 1). AB - Over the last two decades gene therapy has moved from preclinical to clinical studies for many diseases ranging from single gene disorders such as cystic fibrosis and Duchenne muscular dystrophy, to more complex diseases such as cancer and cardiovascular disorders. Gene therapy for severe combined immunodeficiency (SCID) is the most significant success story to date, but progress in many other areas has been significant. We asked 20 leaders in the field succinctly to summarize and comment on clinical gene therapy research in their respective areas of expertise and these are published in two parts in the Progress and Prospect series. PMID- 17909541 TI - Health-related quality of life--an economist's perspective. PMID- 17909542 TI - Does a comprehensive rehydration regimen reduce neurologic complications associated with diabetic ketoacidosis? PMID- 17909543 TI - Should antibiotic treatment replace appendectomy for acute appendicitis? PMID- 17909544 TI - Should triple therapy for Helicobacter pylori eradication be prescribed for 1 week or 2 weeks? PMID- 17909545 TI - Comparing the Cockcroft-Gault and MDRD equations for calculation of GFR and drug doses in the elderly. PMID- 17909547 TI - Primary aldosteronism: the case against screening. PMID- 17909546 TI - Primary aldosteronism: the case for screening. PMID- 17909548 TI - Improved prediction of outcomes in patients with acute intracranial hemorrhage. PMID- 17909549 TI - Can surgical staging affect the postoperative treatment and outcome of patients with endometrial cancer? PMID- 17909550 TI - Does computer-aided detection increase the accuracy of interpretation of mammograms? PMID- 17909551 TI - Is cetuximab active in patients with cisplatin-refractory squamous cell carcinoma of the head and neck? PMID- 17909552 TI - Is treatment with chondroitin efficacious for osteoarthritis? PMID- 17909553 TI - Should infliximab be used to help maintain glucocorticosteroid-induced remission in patients with giant cell arteritis? PMID- 17909554 TI - Should the pT2 tumor classification for renal cell carcinoma be subdivided according to tumor size? PMID- 17909555 TI - Is laparoscopic partial nephrectomy as effective as open partial nephrectomy in patients with renal cell carcinoma? PMID- 17909556 TI - Surgical treatment of intramedullary spinal cord tumors: prognosis and complications. AB - STUDY DESIGN: Retrospective case series. OBJECTIVE: To evaluate our recent treatment strategy for intramedullary spinal cord tumors. SETTING: Department of Orthopaedic Surgery, Keio University, Japan. METHODS: We reviewed 68 cases of intramedullary tumors (ependymoma, 33; astrocytoma, 23; hemangioblastoma, 12), treated surgically between 1994 and 2003. There were 42 males and 26 females whose mean age at the time of surgery was 43 years. The mean follow-up period was 6.2 years. The tumor malignancy grade according to the WHO classification was astrocytoma grade I, 3; grade II, 8 (low-grade: 11 cases); grade III, 10; grade IV, 2 (high-grade: 12 cases). All ependymomas were grade II. Three of the 12 hemangioblastomas were associated with von Hippel-Lindau disease. RESULTS: Total excision was achieved in 90% of the ependymomas and functional improvement was obtained when the preoperative neurological deficit was mild. Approximately 50% of low-grade astrocytomas could be totally excised with favorable survival outcomes, suggesting that total excision should be attempted for low-grade astrocytomas. However, total excision of high-grade tumors was difficult and the functional outcomes were poor. Cordotomy should be considered in patients with a thoracic high-grade astrocytoma. Total resection was possible in 92% of hemangioblastoma, and the functional outcomes were good, however, more attention should be paid for tumors with feeding arteries on the ventral side and for those associated with von Hippel-Lindau disease. CONCLUSIONS: Predictors of good surgical outcome for intramedullary spinal cord tumors were histological grades of the tumors, surgical margins, and neurological status of the patient before surgery. PMID- 17909558 TI - Measurement properties of the CESD scale among individuals with spinal cord injury. AB - BACKGROUND: Depression is common after spinal cord injury (SCI), yet it can be difficult and costly to diagnose. Screening tools such as the Center for Epidemiological Studies Depression scale (CESD) can assist with case identification; however, insufficient knowledge of their measurement properties exists to use them in the SCI population. OBJECTIVES: To assess the reliability and validity of the CESD-20 and the CESD-10. SETTING: Tertiary care centre in Vancouver, British Columbia, Canada. METHODS: A 2-week retest study of 47 individuals with traumatic SCI. Subjects >or=19, who had their SCI for >or=1 year and had American Spinal Injury Association Impairment scale ranking of A or B. Short Form-36 (SF-36) subscales and a visual analogue scale for fatigue (VAS-F) were used to assess validity using Pearson's correlations coefficients. Internal consistency was assessed using Cronbach's alpha, retest reliability was assessed using intraclass correlation coefficients (ICCs) and Bland-Altman plots. Normative data are presented based on key demographic and clinical factors. RESULTS: Cronbach's alpha was 0.91 and 0.86 and retest reliability was ICC=0.87 (95% confidence interval (CI) 0.79-0.93) and ICC=0.85 (95% CI 0.75-0.92) for the CESD-20 and CESD-10, respectively. Minimal bias was evident based on the Bland Altman plots. The strongest correlations were with outcomes representing mental health (r=-0.71), vitality (r=-0.60) and the VAS-F (r=0.57). The weakest correlation was with the physical function score of the SF-36 (r=-0.37). CONCLUSION: The CESD-20 and CESD-10 are quick and easy to use. This study provides evidence in support of the reliability and validity. PMID- 17909557 TI - A review of scales for assessing the risk of developing a pressure ulcer in individuals with SCI. AB - BACKGROUND: Pressure ulcers are a common secondary condition that occur post spinal cord injury (SCI). These ulcers come at tremendous personal and societal cost. There are a number of scales that can be used to identify those who are at risk. OBJECTIVES: This review critically evaluates risk assessment scales designed for identifying and predicting skin ulcers. Specifically, studies on the psychometric properties and utility for individuals with SCI were assessed. METHODS: The MedLine, CINHAL, Embase, HaPI, Psycinfo, Sportdiscus and Cochrane databases were searched to identify studies. To be included, the scale needed to have at least one study, published in a peer-reviewed journal, which examined its psychometric properties with a sample of individuals with SCI. RESULTS: Seven scales were included in this review: Abuzzese, Braden, Gosnell, Norton, SCIPUS, SCIPUS-A and Waterlow. None of the tools reported reliability data with this population. Validity evidence ranged from poor to adequate across scales. Most were readily available, quick to administer and had minimal respondent burden; however, the SCIPUS-A and SCIPUS, two scales developed specifically for individuals with SCI, required laboratory blood testing. CONCLUSION: Although the SCIPUS-A and SCIPUS show promise, utility issues and limited psychometric testing suggest that these tools cannot be recommended at this time. While the Braden scale has the best combined validity and utility evidence, more specific testing with individuals with SCI is required for it and all other scales included in the review. PMID- 17909559 TI - Spasticity outcome measures in spinal cord injury: psychometric properties and clinical utility. AB - STUDY DESIGN: Comprehensive review and systematic analyses. OBJECTIVES: Assess published psychometric evidence for spinal cord injury (SCI) spasticity outcome measures. Considerations about the influence of spasticity on function have also been identified to understand treatment effects and guide service delivery. SETTING: London, Ontario and Vancouver, British Columbia, Canada. METHOD: Review of measures was based on availability of psychometric data, application in clinical settings and evaluated in SCI patients. RESULTS: Ashworth and Modified Ashworth Scales (AS, MAS), Penn Spasm Frequency Scale (PSFS), Spinal Cord Assessment Tool for Spasticity (SCATS), Visual Analogue Scale self-rated scale of spasticity (VAS) and the Wartenberg Pendulum Test (WPT) were included in this review. The most frequently used tools for SCI spasticity measurement include the AS, MAS, PSFS and VAS, of which the latter two are self-report spasticity measures. The SCATS has been partially validated for SCI, but is not widely used. The WPT has been minimally validated despite its use in a large-scale SCI spasticity randomized controlled trial. CONCLUSIONS: Since spasticity is multidimensional, focusing on one or two spasticity outcome measures can misrepresent the extent and influence of spasticity on SCI patients. Different scales measure different aspects of spasticity and individual tools correlate weakly with each other. Spasticity may be better measured with an appropriate battery of tests, including the AS or MAS, along with PSFS. These tools would benefit from further reliability and responsiveness testing. Tools that assess the influence of spasticity on patient activities, participation and quality of life are important, but lacking. PMID- 17909560 TI - Epidural malignant lymphomas of the spine: collected experiences with epidural malignant lymphomas of the spinal canal and their treatment. AB - STUDY DESIGN: Retrospective study of 13 patients treated by the authors. OBJECTIVE: To examine the course of the disease of malignant lymphoma (ML) presenting in the epidural area of the spine. SETTING: Department of Neurosurgery, Third Department of Internal Medicine, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary. SUBJECTS AND METHODS: The epidural presentation in eight patients was heralded by motor signs (paraparesis and plegia), in one by a lesion of the posterior columns of the spinal cord (ataxia), and in three by pain. One patient was free of complaints and symptoms. The affected epidural area was diagnosed previously by myelography and computerized tomography (CT), and later by magnetic resonance (MR), over the course of which the location was verified as thoracic in eight patients, cervical in one, and lumbar in four. The authors recommended surgical intervention in 9 out of 13 cases, in seven cases of Hodgkin's and six cases of non-Hodgkin's lymphoma. Seven patients were treated for recognized manifestations of malignant lymphoma while six were diagnosed by intraoperative-histological examination. RESULTS AND CONCLUSION: The decompression operations for tumors resulted in limited improvement in seven patients (reduction in pain and return of ability to walk). Four patients were not operated on, two of which had significant improvement in their neurological symptoms. Paraparesis remained unchanged in one patient. One patient remained symptom-free. The authors emphasize the importance of interdisciplinary consultation and weighing individual priorities in the indications for operation on epidural ML. PMID- 17909561 TI - Autoimmunity: altered self-N-glycans trigger innate-mediated autoimmunity. PMID- 17909562 TI - Involvement of cannabinoid receptor CB2 in dectin-1-mediated macrophage phagocytosis. AB - Cannabinoid receptors are expressed in macrophages, but little is known of their roles. We here examined their involvement in phagocytosis. The presence of 2 arachidonylglycerol, an endocannabinoid, augmented the phagocytosis of zymosan by mouse macrophages, while the phagocytosis of Escherichia coli, Staphylococcus aureus, apoptotic cells or latex beads remained unaffected. An agonist of the cannabinoid receptors CB1 and CB2 also stimulated the phagocytosis of zymosan. The stimulatory effect of 2-arachidonylglycerol was abolished when phagocytosis reactions were carried out in the presence of an antagonist of CB2 but not of CB1. Furthermore, the phagocytosis of zymosan in the presence of 2 arachidonylglycerol was severely inhibited by the addition of a beta-glucan containing carbohydrate or antibody neutralizing dectin-1, a beta-glucan recognizing phagocytosis receptor. These results suggested that the activation of CB2 in macrophages leads to the stimulation of dectin-1-mediated phagocytosis. PMID- 17909563 TI - Mycobacterium tuberculosis secretory proteins CFP-10, ESAT-6 and the CFP10:ESAT6 complex inhibit lipopolysaccharide-induced NF-kappaB transactivation by downregulation of reactive oxidative species (ROS) production. AB - Mycobacterium tuberculosis (Mtb) causes death of 2-3 million people annually and is considered one of the most successful intracellular pathogens to persist inside the host macrophage. Recent studies have implicated the role of RD-1 region of Mtb genome in the mycobacterial pathogenesis. The role of RD-1-encoded secretory proteins of Mtb in modulation of macrophage function has not been investigated in detail. Here we show that RD-1 encoded two major secretory proteins, namely, culture filtrate protein-10 kDa (CFP-10) and early secreted antigenic target-6 kDa (ESAT-6), and their 1:1 CFP-10:ESAT6 complex inhibit production of reactive oxidative species (ROS) in RAW264.7 cells. These proteins also downregulated the bacterial lipopolysaccharide (LPS)-induced ROS production, which, in turn, downregulated LPS-induced nuclear factor-kappaB (NF-kappaB) p65 DNA-binding activity, as well as inhibited the NF-kappaB-dependent reporter gene (chloramphenicol acetyl transferase) expression in the treated macrophages. Moreover, addition of N-acetyl cysteine, which is a scavenger of ROS, also inhibited LPS-induced reporter gene expression by scavenging the ROS, thereby preventing NF-kappaB transactivation. These studies indicate that the secretory proteins CFP-10, ESAT-6 and the CFP10:ESAT6 complex of Mtb can inhibit LPS induced NF-kappaB-dependent gene expression via downregulation of ROS production. PMID- 17909565 TI - Contemporary clinical management of isolated high-grade prostatic intraepithelial neoplasia. AB - High-grade prostatic intraepithelial neoplasia (HGPIN) is a premalignant lesion associated with increased risk of coexistent cancer or delayed progression to carcinoma. Extended biopsy schemes have improved the ability to rule out concurrent cancers, increased the detection of isolated HGPIN and removed the routine necessity for immediate repeat biopsy. As the natural history of HGPIN is poorly defined, and no non-invasive marker allows monitoring of progression to cancer, routine delayed interval biopsy should be considered in all patients. In this article, we present an overview of the existing literature on HGPIN and a proposed strategy for clinical management. PMID- 17909564 TI - Functional effects of single nucleotide polymorphisms in the coding region of human N-acetyltransferase 1. AB - Genetic variants of human N-acetyltransferase 1 (NAT1) are associated with cancer and birth defects. N- and O-acetyltransferase catalytic activities, Michaelis Menten kinetic constants (K(m) and V(max)) and steady-state expression levels of NAT1-specific mRNA and protein were determined for the reference NAT1*4 and variant human NAT1 haplotypes possessing single nucleotide polymorphisms (SNPs) in the open reading frame. Although none of the SNPs caused a significant effect on steady-state levels of NAT1-specific mRNA, C97T(R33stop), C190T(R64W), C559T (R187stop) and A752T(D251V) each reduced NAT1 protein level and/or N- and O acetyltransferase catalytic activities to levels below detection. G560A(R187Q) substantially reduced NAT1 protein level and catalytic activities and increased substrate K(m). The G445A(V149I), G459A(synonymous) and T640G(S214A) haplotype present in NAT1*11 significantly (P<0.05) increased NAT1 protein level and catalytic activity. Neither T21G(synonymous), T402C(synonymous), A613G(M205V), T777C(synonymous), G781A(E261K) nor A787G(I263V) significantly affected K(m), catalytic activity, mRNA or protein level. These results suggest heterogeneity among slow NAT1 acetylator phenotypes. PMID- 17909566 TI - Sexual function following radical prostatectomy: a prospective longitudinal study of cultural differences between Japanese and American men. AB - We conducted a cross-cultural comparison of the recovery of sexual function and bother during the first 2 years after radical prostatectomy (RP) between American and Japanese men. A total of 275 Japanese and 283 American men who underwent RP alone were prospectively enrolled into longitudinal cohort studies of health related quality of life outcomes. Sexual function and bother (distress) were estimated with English and validated Japanese versions of the UCLA Prostate Cancer Index before RP and 1, 2-3, 4-6, 12, 18 and 24 months after RP. Each subject served as his own control. Japanese men reported lower sexual function scores at baseline, even after adjusted for age, prostate-specific antigen (PSA) and comorbidity (38 vs 61, P<0.001). The two groups had similar baseline sexual bother (70 vs 69, P=0.84). Japanese men had a smaller improvement in sexual function (beta=0.8 vs beta=5.3) and bother (beta=0.2 vs beta=2.9) over time than did the American men postoperatively, after adjusting for baseline score, age, baseline PSA and nerve-sparing. American men were more likely than Japanese men to regain their baseline sexual function by 24 months after surgery (hazard ratio (HR)=1.60; 95% confidence interval (CI)=1.06-2.42). In contrast, American men were less likely than Japanese men to return to baseline sexual bother (HR=0.57; 95% CI=0.44-0.75). This study demonstrates that Japanese and American men experience different patterns of recovery of their sexual function and bother after RP. Ethnicity may be a contributing factor. PMID- 17909568 TI - Adiabatic frequency conversion of optical information in atomic vapor. AB - We experimentally demonstrate a communication protocol that enables frequency conversion and routing of quantum information in an adiabatic and thus robust way. The protocol is based on electromagnetically induced transparency (EIT) in systems with multiple excited levels: transfer and/or distribution of optical states between different signal modes is implemented by adiabatically changing the control fields. The proof-of-principle experiment is performed using the hyperfine levels of the rubidium D1 line. PMID- 17909567 TI - Increased ranging depth in optical frequency domain imaging by frequency encoding. AB - A technique for increasing the ranging depth in optical frequency domain imaging utilizing frequency encoding is presented. Ranging depth is enhanced by using two interferometer reference arms with different path lengths and independent modulation frequencies (25 and 50 MHz). With this configuration, the sensitivity decreases by 6 dB over a depth range of 7 mm, approximately a threefold improvement over the conventional optical frequency domain imaging technique. We demonstrate that the reference arm frequency separation, tuning speed, center wavelength, and instantaneous coherence length determine the signal-to-cross-talk ratio. PMID- 17909569 TI - Stable pulse-compressed acousto-optic Q-switched fiber laser. AB - A novel acousto-optic switch operation by a simple laser-diode pumped acousto optic, Q-switched, ytterbium-doped, double-clad fiber laser is reported. Stable compressed Q-switched sub-40 ns pulses with a beam quality factor (M(2)=2) are achieved at the repetition rate of 1-50 kHz. Q-switched pulses of ~20 microJ pulse energy and 35 ns pulse width are obtained at the repetition rate of 50 kHz. Finally, a reasonable explanation of the novel Q-switched operation is presented. PMID- 17909570 TI - Electro-optically tunable microring resonators on lithium niobate. AB - Electro-optical tuning of a microring resonator fabricated on lithium niobate (LiNbO3) is presented. The device structure, including microring resonator and couplers, is designed in detail and is produced by titanium diffusion on the wet etched LiNbO3 ridge surface. The resonance wavelengths for TM and TE polarizations can be tuned by electro-optic effect. The output characteristics of through port and drop port in the microring resonators are measured, and the effect of applied voltage on the shift of resonant wavelength is discussed. The presented microring resonators have the features of fast tuning speed, high material stability, bidirection wavelength shift, and no heating interference. Realization of such a microring resonator on LiNbO3 makes the utilization of electro-optic tuning and nonlinear effects in the versatile photonic applications of microring resonators achievable. PMID- 17909571 TI - High-pulse-energy actively Q-switched Tm3+-doped silica 2 microm fiber laser pumped at 792 nm. AB - A diode-pumped Q-switched Tm(3+)-doped double-clad silica fiber laser is reported providing average powers of up to 30 W at pulse widths of only 41 ns and repetition rates in the range of 10-125 kHz. Up to 270 microJ pulse energy was produced. Amplified spontaneous emission (ASE) buildup limits the maximum peak power, and the pump power and average output power at the point of ASE induced clamping were found to depend linearly on the repetition rate. PMID- 17909572 TI - Polarization-tunable Yb:KGW laser based on internal conical refraction. AB - Using internal conical refraction, we demonstrate an Yb:KGW laser where the polarization state can be arbitrarily altered without any additional cavity components. The extinction ratio can also be altered between 1:1 and 40:1. The maximum output power achieved was 8.6 W at a slope efficiency of 60.5% with respect to incident pump power. This equals the power performance of a standard Yb:KGW laser used for reference. PMID- 17909573 TI - Pump-probe analysis of the cross-phase modulation degradation induced by 10 Gbit/s amplitude-shift-keyed signals on 40 Gbit/s DPSK signals. AB - A study of the cross-phase modulation (XPM) degradation of differential-phase shift-keyed (DPSK) signals due to amplitude-shift-keyed signals is performed using pump-probe simulation. Approximate expressions for the contributions of the XPM-induced intensity and phase modulation to the electrical current fluctuations at the differential-phase-exchange-keyed receiver are presented. It is shown that, unlike prior works and similar to intensity-modulated signals, the contribution of XPM-induced intensity modulation is dominant in systems using standard fiber or high residual dispersion. PMID- 17909574 TI - Dielectric microconcentrators for efficiency enhancement in concentrator solar cells. AB - Metal fingers typically cover more than 10% of the active area of concentrator solar cells. Microfabricated dielectric optical designs that can completely eliminate front contact shading losses are explored. Essentially no microconcentrator optical losses need be incurred, series resistance losses can be reduced, and net efficiency gains of roughly 15% (relative) are realistic. PMID- 17909575 TI - Room-temperature continuous-wave operation of an external-cavity quantum cascade laser. AB - Room-temperature, continuous-wave operation of an external-cavity quantum cascade laser (EC-QCL) is reported. Single-mode tuning range of 120 cm(-1) was achieved, from 7.96 to 8.84 microm. The gain chips utilized are based on the bound to continuum design and were fabricated as buried heterostructure lasers. Gap-free tuning (mode hops only on the external-cavity modes) is demonstrated for an antireflection-coated laser, just by grating rotation. The EC-QCL was implemented in a Littrow setup and an average power of 1.5 mW was obtained at 20 degrees C, while a peak power of 20 mW was obtained for a modified Littrow setup with the back extraction of light. PMID- 17909576 TI - Nonlinear bouncing of nonlocal spatial solitons at the boundaries. AB - We investigate, both experimentally and theoretically, the interaction of (2D+1) spatial solitons with the boundaries of a nonlinear medium in the presence of nonlocality. We demonstrate power-dependent nonlinear repulsion at the boundaries, in quantitative agreement with the model predictions. PMID- 17909577 TI - Laser Doppler imaging through tissues phantoms by using self-mixing interferometry with a laser diode. AB - Laser Doppler imaging has been widely used for the evaluation of cutaneous blood flow. We report on how the self-mixing interferometry configuration with a laser diode is explored for what is believed to be the first time to generate flow maps. The experiment was carried out by sensing the laser intensity power spectrum at each pixel as the laser was scanned over a model that mimics the properties of skin and circulating blood. PMID- 17909578 TI - Low-loss, low-cross-talk crossings for silicon-on-insulator nanophotonic waveguides. AB - We present compact crossings for silicon-on-insulator photonic wires. The waveguides are broadened using a 3 microm parabolic taper in each arm. By locally applying a lower index contrast using a double-etch technique, loss of confinement is reduced and 97.5% transmission (-1.7 dB) is achieved with only -40 dB cross talk. PMID- 17909579 TI - Narrow linewidth, 100 W cw Yb3+-doped silica fiber laser with a point-by-point Bragg grating inscribed directly into the active core. AB - We report on the power scaling to 103 W of a 1.1 microm continuous-wave Yb(3+) doped silica fiber laser incorporating a point-by-point (PbP) fiber-Bragg grating inscribed directly into the active core using 800 nm femtosecond laser pulses. The spectrum of the laser exhibited a narrow linewidth that broadened to 260 pm at 103 W. The output was frequency doubled using an 11 mm long periodically poled MgO:LiNbO3 crystal to generate 2.1 W of green with an internal conversion efficiency of 10% at high power and 0.81%/W at low power. PMID- 17909580 TI - Polymeric saturable-absorber mirror for passive Q-switching of a laser-diode pumped Nd3+:YVO4 microchip laser. AB - A polymeric multilayered mirror doped with a saturable dye worked as a passive Q switch of a laser-diode-pumped Nd(3+):YVO4 microchip laser. The multilayered mirror consisted of alternately spin-coated layers of polyvinylcarbazole (PVK) and cellulose acetate on a glass substrate. A dye of bis (dimethylaminodithiobenzil)-nickel was doped in one of the layers of PVK, providing the repetitively Q-switched pulses. The pulse width and repetition rate were 4 ns and 68 kHz, respectively, for a laser-cavity length of 5 mm, and the average and the peak power were 40 mW and 156 W, respectively, for the highest pump power of 435 mW. The dependence of the Q-switched characteristics on the pump power and on the concentration of the doped dye is described. PMID- 17909581 TI - Measurement of the absolute energy level and hyperfine structure of the 87Rb 4D5/2 state. AB - We have measured the absolute frequency of the excited state transition 5P(3/2) 4D(5/2) in a (87)Rb atom with a femtosecond frequency comb, utilizing the recently developed spectroscopic technique of the double resonance optical pumping method. The absolute energy level of the 4D(5/2) state is determined by measuring the absolute frequency of the 5S(1/2)-5P(3/2) transition simultaneously. The hyperfine structure constants of the 4D(5/2) state are obtained by using the measured frequency. The magnetic dipole constant, A, is determined to be (-16.747+/-0.010) MHz with an uncertainty reduced 60-fold compared with a previous result. The electric quadrupole constant, B, is determined, for what is to our knowledge the first time, to be (4.149+/-0.059) MHz. PMID- 17909582 TI - Wavelength filtering characteristics of Solc filter based on Ti:PPLN channel waveguide. AB - We have analyzed the Solc filtering characteristics in a periodically poled Ti:LiNbO3 (Ti:PPLN) multimode waveguide. The single- and dual-wavelength filtering were achieved under the optimized guiding condition for the TEM(00) like mode and two mode (TEM(00)- and TEM(01)-like mode), respectively. The full width at half-maximum of the filter was about 0.21 nm at both guiding conditions. We found that the origin of two peaks of the dual-wavelength Solc filter in the two-mode guiding condition is the different effective refractive index between the TEM(00)- and TEM(01)-like modes. The wavelength difference of two peaks is about 0.8 nm at room temperature. PMID- 17909583 TI - Supercontinuum and third-harmonic generation accompanying optical filamentation as first-order scattering processes. AB - It is demonstrated numerically that the supercontinuum generation and third harmonic generation that accompany optical filamentation in nonlinear dispersive bulk media can be described as first-order scattering processes akin to the first Born approximation. In particular, for an incident ultrashort pulse the angularly resolved spectrum of the transmitted pulse is shown to be accurately determined using first-order scattering of the incident field from the nonlinearly modified refractive index due to the optical filament. Thus, although an optical filament is a highly nonlinear object, the accompanying supercontinuum generation and third-harmonic generation are driven parametrically by the filament and have negligible back action upon it. PMID- 17909584 TI - Spontaneous growth of Raman Stokes and anti-Stokes waves in fibers. AB - Using the quantum theory of light, we derive general analytical expressions of Stokes and anti-Stokes spectral photon-flux densities that are spontaneously generated by a single monochromatic pump wave propagating in a single-mode optical fiber. We validate our results by comparing them with experimental data. Limiting cases corresponding to interesting physical situations are discussed. PMID- 17909585 TI - Modified spectrum autointerferometric correlation (MOSAIC) for single-shot pulse characterization. AB - A method for generation of the modified spectrum autointerferometric correlation that allows single-shot pulse characterization is demonstrated. A sensitive graphical representation of the ultrashort pulse phase quality is introduced that delineates the difference between the presence of temporal and spectral phase distortions. Using these schemes, full-field reconstruction of ultrashort laser pulses is obtained in real time using an efficient iterative technique. PMID- 17909586 TI - Enhanced optical absorption for photovoltaics via excitation of waveguide and plasmon-polariton modes. AB - We study theoretically the mechanisms by which optical absorption is enhanced in an optical nanostructure consisting of a slab waveguide, made of a-Si:H, sandwiched between a periodic array of metallic nanowires and a substrate, both made of Au. We demonstrate that for the TM polarization the optical absorption in the slab waveguide can be enhanced by almost an order of magnitude by the excitation of plasmon modes, whereas for both the TM and TE polarizations the grating-induced excitation of slab waveguide modes leads to more than twofold enhancement of the optical absorption. PMID- 17909587 TI - Multikilohertz optical parametric chirped pulse amplification in periodically poled stoichiometric LiTaO3 at 1235 nm. AB - We demonstrate, for the first time to our knowledge, multikilohertz operation of a double-stage optical parametric chirped pulse amplifier with periodically poled stoichiometric LiTaO3 crystals, seeded by a homemade Cr:forsterite oscillator operating at 1235 nm. The repetition rate of the amplifier could easily be tuned without the use of electro-optic modulators by using a repetition-rate-tunable kilohertz pump laser operating at 532 nm and a time-synchronization unit. Amplified total (signal+idler) energies of 55.9 and 36.2 microJ were achieved at 1 and 5 kHz, respectively. After recompression, the pulse width of amplified idler pulses at 1235 nm amounted to 530 fs. PMID- 17909588 TI - Tunable single-mode fiber-VCSEL using an intracavity polymer microlens. AB - We report a tunable, single-mode vertical cavity surface-emitting laser (VCSEL) format suitable for array operation, power scaling, fiber coupling, and operation in isolated environments such as those required by atom optics. The devices are fiber VCSELs, consisting of a semiconductor gain and mirror structure separated from a mirror-coated optical fiber by an air (or vacuum) gap. The gain structure has polymer microlenses fabricated on its surface, of characteristics suitable to focus the oscillating mode on both cavity mirrors, ensuring stable fundamental mode emission and high fiber coupling efficiency. We demonstrate such devices in continuous-wave operation at 1.03 microm at room temperature, with a single-mode tuning range of 13 nm, laser threshold as low as 2.5 mW, and a maximum fiber coupled output power of 10 mW. PMID- 17909589 TI - Surface plasmon resonance biosensor based on electro-optically modulated phase detection. AB - A novel electro-optically modulated surface plasmon resonance (EOMSPR) biosensor based on phase detection by collinear heterodyne interferometry is presented. Its operation is by means of modulating the SPR-induced phase variation by electro optic effect. Because the relation between the phase change and the applied voltage is highly sensitive to analyte properties, the linear regression slope of this relation can be used in the analyte detection. The detection characteristics of this EOMSPR biosensor are measured, and their dependences on waveguide width and gold-film thickness are discussed. This novel EOMSPR biosensor has attractive features, such as compactness, resolution tunability, and noise reduction. PMID- 17909591 TI - High-power and high-temperature THz quantum-cascade lasers based on lens-coupled metal-metal waveguides. AB - A metal-metal waveguide quantum cascade laser with an abutted silicon hyperhemispherical lens is demonstrated at ~4.1 THz. The device produced 145 mW of peak pulsed power at 5 K with a wall-plug power efficiency of 0.7%, lasing up to a maximum operating temperature of 160 K. The far-field beam pattern has a full width at half-maximum value of ~4.8 degrees in the H plane. The same device produced ~26 mW of peak power using a Winston cone instead of a lens, lasing up to 165 K. The large increase in output power is mainly attributed to an increase in collection efficiency. PMID- 17909590 TI - Flux vector formulation for photon propagation in the biological tissue. AB - We present a generalized delta-Eddington phase function to simplify the radiative transfer equation to an integral equation with respect to the photon flux vector. The solution of the integral equation is highly accurate to model the photon propagation in the biological tissue over a broad range of optical parameters, especially in the visible light spectrum where the diffusion approximation breaks down. The methodology is validated in the Monte Carlo simulation and can be applied in various optical imaging applications. PMID- 17909592 TI - Enhancement of the evanescent field using polymer waveguides fabricated by deep UV exposure on mesoporous silicon. AB - Polymer integrated reverse symmetry waveguides on porous silicon substrate fabricated by using deep ultraviolet radiation in poly(methyl methacrylate) are presented. The layer sequence and geometry of the waveguide enable an evanescent field extending more than 3 microm into the upper waveguide or analyte layer, enabling various integrated optical devices where large evanescent fields are required. The presented fabrication technique enables the generation of defined regions where the evanescent field is larger than in the rest of the waveguide. This technology can improve the performance of evanescent-wave-based waveguide devices. PMID- 17909593 TI - Enhanced weak-signal sensitivity in two-photon microscopy by adaptive illumination. AB - We describe a technique to enhance both the weak-signal relative sensitivity and the dynamic range of a laser scanning optical microscope. The technique is based on maintaining a fixed detection power by fast feedback control of the illumination power, thereby transferring high measurement resolution to weak signals while virtually eliminating the possibility of image saturation. We analyze and demonstrate the benefits of adaptive illumination in two-photon fluorescence microscopy. PMID- 17909594 TI - Spectral imaging system for scaling the power spectrum of optical waveforms. AB - We propose and analyze a new photonic system to change the scale of the optical power spectrum of an ultrashort pulse. Our device corresponds to the dual (spectral domain) configuration of a temporal imaging system. It is configured with two time lenses separated by a linear first-order dispersive medium. The specific relation between the chirp rates and the first-order dispersion coefficient is obtained. The main practical constraints are discussed, and the system performance is tested by means of a numerical simulation. PMID- 17909595 TI - Off-axis behavior of an infrared meander-line waveplate. AB - An infrared meander-line waveplate has been modeled and measured over the 8 to 12 microm spectral band in terms of its differential phase delay, axial ratio of the output polarization ellipse, and power throughput for angles of incidence between 0 degrees and 60 degrees. The study has been performed for planes of incidence parallel and perpendicular to the meander-line axis. The main significance is that the phase delay remains almost unaffected by the angle of incidence. Infrared meander-line retarders can thus be used well beyond the paraxial range as in low-f/# optical systems and in non-normal-incidence applications. PMID- 17909596 TI - TOMBO sensor with scene-independent superresolution processing. AB - One approach to flat sensor design is to use a lenslet array to form multiple subimages of a scene and then combine the subimages to recover a fully sampled image by using a superresolution algorithm. Previously, superresolution image assembly has been based on information derived from the observed scene. For lenslet arrays, we propose a new scene-independent approach based only on known imager properties in which relative subimage shifts are accurately estimated with a calibration procedure using point source imaging. Thus, the relative resolution enhancement provided by the scene-independent superresolution algorithm is impervious to changes in subimage content, contrast, sharpness, and noise. PMID- 17909597 TI - Chirped coherent anti-Stokes Raman scattering as a high-spectral- and spatial resolution microscopy. AB - Coherent anti-Stokes Raman scattering (CARS) microscopy is a promising tool for chemically selective imaging based on molecular vibrations. While CARS is currently used as a biological imaging tool, many variations are still being developed, perhaps the most important being multiplex CARS microscopy. Multiplex CARS has the advantage of comparing images based on different molecular vibrations without changing the excitation wavelengths. Here we demonstrate both high-spectral- and spatial-resolution multiplex CARS imaging of polymer films, using a simple scheme for chirped CARS with a spectral bandwidth of 300 cm(-1). PMID- 17909598 TI - Efficient fiber-to-waveguide coupling through the vertical leakage from a microring. AB - A fiber-to-waveguide coupler is proposed to efficiently couple light from a single-mode fiber into a submicrometer semiconductor waveguide for integration with optoelectronic circuits. A microring with a specific refractive index is designed on the top of the semiconductor waveguide. The gradual vertical leakage from the microring forms steady coupling into the semiconductor waveguide. Coupling efficiency up to 93% is demonstrated using the three-dimensional finite difference time-domain method. A tapered-waveguide or microring structure can be used to convert the lateral-mode size for coupling light into a single-mode semiconductor waveguide. PMID- 17909599 TI - Standing-wave nonlinear optics in an integrated semiconductor microcavity. AB - We present a concept of standing-wave optical frequency conversion in dispersive microcavities theoretically and experimentally, allowing efficient ultracompact nonlinear photonics. We developed a time-dependent model, incorporating the dispersion into the structure of the spatial cavity modes, where the conversion efficiency is enhanced by the optimization of a nonlinear cavity mode overlap. We designed and fabricated integrated double-resonance semiconductor microcavities for standing-wave second-harmonic generation. The measured efficiency exhibits a significant maximum near the cavity resonance owing to the intracavity power enhancement and the dispersion-induced wavelength detuning effect on the mode overlap, in good agreement with our theoretical predictions. PMID- 17909600 TI - Time-variant signal encryption by lensless dual random phase encoding applied to fiber optic links. AB - A lensless dual random phase encoding technique in the temporal domain is proposed and analyzed to evaluate its potential application for secure data transmission, mainly for short-haul fiber optic links. The different signal broadening effects produced by each stage of the encoding process in both time and frequency domains are analyzed by using the Wigner distribution function to take into account the fiber's multiplexing capabilities. Thus, quasi-white noise with a well-defined bandwidth is used in the encoding process to limit the bandwidth of the encrypted signal. Numerical simulations revealed good system performance, indicating that this multiplexing encryption method could be a good alternative to other well-established techniques. PMID- 17909601 TI - Cavity-enhanced similariton Yb-fiber laser frequency comb: 3 x 10(14) W/cm2 peak intensity at 136 MHz. AB - We report on a passive cavity-enhanced Yb-fiber laser frequency comb generating 230 MW of peak power (3 kW of average power) at a 136 MHz pulse repetition rate. The intracativy peak intensity of 3 x 10(14) W/cm2 for the 95 fs pulse is sufficient to ionize noble gases, such as Xe, Kr, or Ar. The laser system is based on a mode-locked Yb-fiber similariton oscillator in conjunction with a cladding-pumped chirped-pulse fiber amplifier. After recompression, 75 fs duration pulses at a 13.1 W average power are obtained. These pulses are then coherently added inside a passive ring cavity by controlling the fiber oscillator's pulse repetition rate and carrier-envelope offset frequency. This system is well suited for studying high-field phenomena at very high pulse repetition rates. PMID- 17909602 TI - High-power and highly efficient Tm3+-doped silica fiber lasers pumped with diode lasers operating at 1150 nm. AB - An output power of 1.74 W at 2.03 microm was generated at a slope efficiency of 51% when a double-clad Tm(3+)-doped silica fiber laser was pumped with high-power 1150 nm diode lasers. Pump excited state absorption from the upper laser level populates higher energy levels allowing cross relaxation to repopulate the upper laser level at a quantum efficiency greater than unity and to limit losses relating to additional pump excited state absorption. The output power was scaled to 4.77 W when both ends of the fiber were pumped. PMID- 17909603 TI - Low-noise, tunable diode laser for ultra-high-resolution spectroscopy. AB - We demonstrate the excellent spectral properties of a diode laser setup that combines good tunability with superb short-term frequency stability and controllability. It is based on merging two concepts, the diode laser with resonant optical feedback and the grating stabilized diode laser. To characterize the short-term performance we beat two essentially identical diode lasers and find a short-term linewidth of ~11 kHz. Phase locking between these lasers is achieved with a servo bandwidth as small as 46 kHz, although an analog phase detector is used that requires subradian residual phase error. Despite small phase error detection range and small servo bandwidth, cycle-slip-free phase locking is accomplished for typically many 10 min, and the optical power is essentially contained in a spectral window of less than 20 mHz relative to the optical reference. Due to the excellent performance this laser concept is well suited for atomic or molecular coherence experiments, which require phase locking of different lasers to each other, and as part of a flywheel for optical clocks. PMID- 17909604 TI - Effects of the carrier-envelope phase of chirped laser pulses in the multiphoton ionization regime. AB - By solving the time-dependent Schrodinger equation for the Cs atom, we find that, as long as the spectral bandwidth is sufficiently broad, the asymmetry of photoelectron ejection is strongly phase dependent and persists even when the chirped pulse duration becomes more than several cycles. The asymmetry survives even after the angle integration over the hemisphere, implying that the detection efficiency can be significantly improved. This counterintuitive and robust finding provides a simple way to measure the phase for both transform-limited and chirped pulses. PMID- 17909606 TI - High-power terahertz-wave generation using DAST crystal and detection using mid infrared powermeter. AB - The exact power output of a table-top-sized terahertz (THz)-wave source using a nonlinear optical process has not been clarified because detectors for these experiments [Si bolometer, deuterated triglycine sulfate (DTGS), etc.] are not calibrated well. On the other hand, powermeters for the mid-infrared (mid-IR) region are well established and calibrated. We constructed a high-power dual wavelength optical parametric oscillator with two KTP crystals as a light source for difference frequency generation. The obtained powers of dual waves were 21 mJ at ~1300 nm, ten times higher than that of the previous measurement. The device provides high-power THz-wave generation with ~100 times greater output power than that reported in previous works. A well-calibrated mid-IR powermeter at ~27 THz detected the generated THz wave; its measured energy was 2.4 microJ. Although the powermeter had no sensitivity in the lower-frequency range (below 20 THz), the pulse energy at such a low-frequency region was estimated in reference to the output spectrum obtained using a DTGS detector: the energy would be from about the submicrojoule level to a few microjoules in the THz-wave region. PMID- 17909605 TI - Guiding in the visible with "colorful" solid-core Bragg fibers. AB - We report on the fabrication and characterization of solid-core all-polymer Bragg fibers consisting of a large-diameter polymethyl methylacrylate (PMMA) core surrounded by 50 alternating PMMA/Polystyrene (PS) polymer layers. By modifying the reflector layer thickness we illustrate that bandgap position can be adjusted at will in the visible. Moreover, such fibers are intensely colored in both the transmission and the outside reflection modes. Potential applications of such fibers are discussed. PMID- 17909607 TI - Femtosecond laser erasing and rewriting of self-organized planar nanocracks in fused silica glass. AB - Tightly focused, linearly polarized, femtosecond laser radiation can produce highly birefringent nanograting structures inside fused silica glass. Here we report that when the polarization direction of the femtosecond light is changed, old nanogratings are erased and simultaneously replaced with new ones whose orientation is solely determined by the polarization of the rewrite beam. We also show that these volume nanogratings can be rewritten 1000 times with little degradation in their quality. PMID- 17909608 TI - Multispectral photoacoustic imaging of fluorochromes in small animals. AB - Fluorochromes have become essential reporter molecules in biological research. We show that the depth-resolved distribution of fluorochromes in small animals can be imaged with 25 fmol sensitivity and 150 microm spatial resolution by means of multispectral photoacoustic imaging. The major advantage of the multispectral approach is the sensitive differentiation of chromophores and fluorochromes of interest based on self-reference measurements, as evidenced in this study by resolving a commonly used fluorochrome (Alexa Fluor 750) in mouse. The suggested method is well suited for enhancing visualization of functional and molecular information in vivo and longitudinally. PMID- 17909609 TI - Electrically tunable slow and fast lights in a quantum-dot semiconductor optical amplifier near 1.55 microm. AB - We have demonstrated both slow light in the absorption regime and fast light in the gain regime of a 1.55 microm quantum-dot semiconductor optical amplifier at room temperature. The theory with coherent population oscillations and four-wave mixing effects agrees well with the experimental results. We have observed a larger phase delay at the excited state than that at the ground state transition, likely due to the higher gain and smaller saturation power of the excited state. PMID- 17909610 TI - Intense femtosecond extreme ultraviolet pulses by using a time-delay-compensated monochromator. AB - Extreme-ultraviolet pulses, produced by high-order harmonic generation, have been spectrally selected by a time-delay-compensated grating monochromator. Temporal characterization of the harmonic pulses has been obtained using cross-correlation method: pulses as short as 8 fs, with high photon flux, have been measured at the output of the monochromator. PMID- 17909611 TI - Cascaded dynamic eigenstates of polarization analysis for piezoelectric polarization control. AB - We propose the cascaded dynamic eigenstates (DESs) of polarization to analyze multicomponent polarization control (PC) devices, and achieve the analytical expression of output state of polarization (SOP) as a function of voltage for piezoelectric polarization control (PPC). By measuring the DES at the output port of the device, the prestage DESs will rotate around subsequent ones. Experimental results in PPC confirm the validity of our analysis. The average error of our theoretical output SOP is 1.23 degrees, and the SOP response time is ~10 micros, which is promising to realize a quasi-open-loop high-speed PC. PMID- 17909613 TI - Standardisation--the theory and the practice. PMID- 17909614 TI - Traceability, reference systems and result comparability. AB - The standardisation of measurements is of high priority in laboratory medicine, its purpose being to achieve closer comparability of results obtained using routine measurement procedures. At present, there is international cooperation in developing reference measurement systems (reference methods, reference materials, and reference laboratory networks) for analytes of clinical significance. These reference systems will reduce, wherever possible, measurement uncertainty and promote the comparability of results. The implementation of measurement traceability through the reference system provides one of the most important tools that supports the standardisation process in laboratory medicine. It aims to achieve result comparability regardless of the measurement procedure (test kit) and the clinical laboratory where analyses are carried out. The aim of this review is to discuss some concepts related to the achievement of standardisation by the implementation of a metrologically-correct measurement system and to provide some examples that illustrate the complexity of this approach and the impact of these activities on patient care. PMID- 17909612 TI - The changing role of the clinical laboratory in the investigation of polycystic ovarian syndrome. AB - In this review we outline clinical features, presentation and pathogenesis of polycystic ovarian syndrome (PCOS), treatment objectives and therapeutic options. We focus on and outline the changing role of the clinical laboratory in diagnosis and treatment of this condition. We also review recent information on the involvement of insulin resistance in the syndrome. We provide some explanation for confusion over the selection of the best hormone measurements for diagnosis. Finally, we outline the best current and future laboratory support for this common condition in young women. PMID- 17909615 TI - The Joint Committee for Traceability in Laboratory Medicine (JCTLM): a global approach to promote the standardisation of clinical laboratory test results. AB - Clinical laboratories are moving towards global standardisation to produce equivalent test results across space and time. Standardisation allows use of evidence-based medicine, eliminates the need of method-specific reference intervals, decision levels and cut-offs, and can be achieved by application of metrological principles. For example, in vitro diagnostics (IVD) manufacturers can make kit calibrators traceable to internationally recognised reference materials and reference methods. The first step towards standardisation is to identify appropriate reference materials and methods. This has been undertaken by a new international consortium, the Joint Committee for Traceability in Laboratory Medicine (JCTLM), formed in 2002. It brings together experts representing the clinical laboratory profession, government agencies, and manufacturers, to promote international comparability, reliability, and equivalence of measurement results in clinical laboratories for the purpose of improving healthcare. Through the efforts of the JCTLM, manufacturers are able to assign values to kit calibrators with consistency using appropriate higher order reference materials and methods, and traceability flowcharts, according to ISO Standards to ensure accuracy of test results and to promote assay performance harmonisation. Users of assay kits can assess suitability of calibrators on the basis of acceptable reference materials and/or methods identified by the JCTLM. The JCTLM exemplifies the dynamic nature of clinical laboratory medicine, the inherent spirit of cooperation among professionals in this scientific field, and the international desire to strive for the highest level of clinical laboratory practice for the benefit of patients. PMID- 17909618 TI - Translating medical science around the world. PMID- 17909616 TI - Prerequisites for use of common reference intervals. AB - The theory of reference values was developed more than 30 years ago, but its application in most clinical laboratories is still incomplete today. This is for several reasons, the most relevant ones being the lack of standardisation of the analytical methods, resulting in method-dependent values, and the difficulty in recruiting the proper number of reference subjects for establishment of reference intervals. With the recent progress in method standardisation the first problem is reducing while the second can be addressed optimally via multicentre collaborative studies that aim to establish common reference intervals. To be effective this approach requires the following prerequisites: 1) the existence of a reference measurement system for the analyte; 2) field methods producing results traceable to the reference system; and 3) a carefully planned multicentre reference interval study. Such a procedure will produce results traceable to the reference measurement system for a large number of reference subjects, under controlled pre-analytical conditions. It will also enable a better understanding of the various sources of population variability, if there is the need for partitioning of a reference interval or if there are any limitations to adopting the established reference intervals on a national or global scale. Once reference intervals are determined, clinical laboratories can adopt a common reference interval provided: 1) the population that the laboratory services is similar to the one studied; 2) methods producing traceable results are used; and 3) analytical quality is within defined targets of precision and bias. Moreover, some validation of the interval using a small sample of reference individuals from the laboratory's population is advisable. PMID- 17909620 TI - Nipping at cardiac remodeling. AB - Much of the mortality following myocardial infarction results from remodeling of the heart after the acute ischemic event. Cardiomyocyte apoptosis has been thought to play a key role in this remodeling process. In this issue of the JCI, Diwan and colleagues present evidence that Bnip3, a proapoptotic Bcl2 family protein, mediates cardiac enlargement, reshaping, and dysfunction in mice without influencing infarct size. PMID- 17909619 TI - New molecularly targeted therapies for lung cancer. AB - Lung cancer is the leading cause of cancer death worldwide. The disease is particularly difficult to detect, and patients often present at an advanced stage. Current treatments have limited effectiveness, and unfortunately, the prognosis remains poor. Recent insights into the molecular pathogenesis and biologic behavior of lung cancer have led to the development of rationally designed methods of early detection, prevention, and treatment of this disease. This article will review the important clinical implications of these advances, with a focus on new molecularly targeted therapies currently in development. PMID- 17909622 TI - Role for HLA in susceptibility to infectious mononucleosis. AB - Factors involved in determining whether infectious mononucleosis occurs after primary EBV infection may include age, dose of virus received, and various genetic markers. A study by McAulay and colleagues reported in this issue of the JCI shows that the presence of certain HLA class I alleles correlates with the incidence and severity of infectious mononucleosis. These same HLA alleles are also risk factors for EBV-associated Hodgkin lymphoma (HL), supporting recent epidemiology that indicates that a history of infectious mononucleosis predisposes to HL. Recent studies suggest that an EBV vaccine might help to prevent infectious mononucleosis, and further development of this should now be considered. PMID- 17909621 TI - Obesity and the beta cell: lessons from leptin. AB - In this issue of the JCI, Morioka et al. report on mice with a whole-pancreas knockout of the leptin receptor that exhibit improved glucose tolerance due to enhanced insulin secretion . At first glance, their findings are very different from those reported in another recent study in which beta cell-specific and hypothalamic knockout of the same gene caused obesity and impaired beta cell function. The differences, which are understandable when one considers the body weights of the animals studied, provide new insight into the links among insulin, leptin action, and beta cell function. PMID- 17909623 TI - Overstaying their welcome: defective CX3CR1 microglia eyed in macular degeneration. AB - Age-related macular degeneration (AMD), the most common cause of blindness in the elderly, is characterized by degeneration of the macula and can lead to loss of fine color vision. Alterations in inflammatory and immune system pathways, which arise from genetic differences, predispose individuals to AMD. Yet the mechanism of disease progression with respect to inflammation is not fully understood. In this issue of the JCI, the study by Combadiere and colleagues shows that CX3C chemokine receptor 1-deficient (CX3CR1-deficient) mice have abnormal microglia that accumulate beneath the retina and contribute to the progression of AMD. PMID- 17909624 TI - Why targeted therapy hasn't worked in advanced cancer. AB - In this issue of the JCI, Nissen et al. report that a reciprocal interaction exists between the growth factors FGF2 and PDGF-BB, causing tumors to exhibit increased angiogenesis and metastatic potential. Both FGF2 and PDGF-BB signal through tyrosine kinase receptors, which have been the target of tyrosine kinase inhibitors for cancer therapies. These inhibitors are usually small molecules that inhibit the kinase activity of a receptor or nonreceptor tyrosine kinase, preventing downstream signaling. The results of this study shed light on why tyrosine kinase inhibitors have been useful for the treatment of only a small number of advanced cancers. Currently, a major focus of pharmaceutical companies is to develop ever more potent and specific tyrosine kinases. The results presented here suggest that this approach may not be successful. PMID- 17909625 TI - Angiogenic factors FGF2 and PDGF-BB synergistically promote murine tumor neovascularization and metastasis. AB - Tumors produce multiple growth factors, but little is known about the interplay between various angiogenic factors in promoting tumor angiogenesis, growth, and metastasis. Here we show that 2 angiogenic factors frequently upregulated in tumors, PDGF-BB and FGF2, synergistically promote tumor angiogenesis and pulmonary metastasis. Simultaneous overexpression of PDGF-BB and FGF2 in murine fibrosarcomas led to the formation of high-density primitive vascular plexuses, which were poorly coated with pericytes and VSMCs. Surprisingly, overexpression of PDGF-BB alone in tumor cells resulted in dissociation of VSMCs from tumor vessels and decreased recruitment of pericytes. In the absence of FGF2, capillary ECs lacked response to PDGF-BB. However, FGF2 triggers PDGFR-alpha and -beta expression at the transcriptional level in ECs, which acquire hyperresponsiveness to PDGF-BB. Similarly, PDGF-BB-treated VSMCs become responsive to FGF2 stimulation via upregulation of FGF receptor 1 (FGFR1) promoter activity. These findings demonstrate that PDGF-BB and FGF2 reciprocally increase their EC and mural cell responses, leading to disorganized neovascularization and metastasis. Our data suggest that intervention of this non-VEGF reciprocal interaction loop for the tumor vasculature could be an important therapeutic target for the treatment of cancer and metastasis. PMID- 17909626 TI - Inhibition of ischemic cardiomyocyte apoptosis through targeted ablation of Bnip3 restrains postinfarction remodeling in mice. AB - Following myocardial infarction, nonischemic myocyte death results in infarct expansion, myocardial loss, and ventricular dysfunction. Here, we demonstrate that a specific proapoptotic gene, Bnip3, minimizes ventricular remodeling in the mouse, despite having no effect on early or late infarct size. We evaluated the effects of ablating Bnip3 on cardiomyocyte death, infarct size, and ventricular remodeling after surgical ischemia/reperfusion (IR) injury in mice. Immediately following IR, no significant differences were observed between Bnip3(-/-) and WT mice. However, at 2 days after IR, apoptosis was diminished in Bnip3(-/-) periinfarct and remote myocardium, and at 3 weeks after IR, Bnip3(-/-) mice exhibited preserved LV systolic performance, diminished LV dilation, and decreased ventricular sphericalization. These results suggest myocardial salvage by inhibition of apoptosis. Forced cardiac expression of Bnip3 increased cardiomyocyte apoptosis in unstressed mice, causing progressive LV dilation and diminished systolic function. Conditional Bnip3 overexpression prior to coronary ligation increased apoptosis and infarct size. These studies identify postischemic apoptosis by myocardial Bnip3 as a major determinant of ventricular remodeling in the infarcted heart, suggesting that Bnip3 may be an attractive therapeutic target. PMID- 17909627 TI - Disruption of leptin receptor expression in the pancreas directly affects beta cell growth and function in mice. AB - Obesity is characterized by hyperinsulinemia, hyperleptinemia, and an increase in islet volume. While the mechanisms that hasten the onset of diabetes in obese individuals are not known, it is possible that the adipose-derived hormone leptin plays a role. In addition to its central actions, leptin exerts biological effects by acting in peripheral tissues including the endocrine pancreas. To explore the impact of disrupting leptin signaling in the pancreas on beta cell growth and/or function, we created pancreas-specific leptin receptor (ObR) KOs using mice expressing Cre recombinase under the control of the pancreatic and duodenal homeobox 1 (Pdx1) promoter. The KOs exhibited improved glucose tolerance due to enhanced early-phase insulin secretion, and a greater beta cell mass secondary to increased beta cell size and enhanced expression and phosphorylation of p70S6K. Similar effects on p70S6K were observed in MIN6 beta cells with knockdown of the ObR gene, suggesting crosstalk between leptin and insulin signaling pathways. Surprisingly, challenging the KOs with a high-fat diet led to attenuated acute insulin secretory response to glucose, poor compensatory islet growth, and glucose intolerance. Together, these data provide direct genetic evidence, from a unique mouse model lacking ObRs only in the pancreas, for a critical role for leptin signaling in islet biology and suggest that altered leptin action in islets is one factor that contributes to obesity-associated diabetes. PMID- 17909628 TI - CX3CR1-dependent subretinal microglia cell accumulation is associated with cardinal features of age-related macular degeneration. AB - The role of retinal microglial cells (MCs) in age-related macular degeneration (AMD) is unclear. Here we demonstrated that all retinal MCs express CX3C chemokine receptor 1 (CX3CR1) and that homozygosity for the CX3CR1 M280 allele, which is associated with impaired cell migration, increases the risk of AMD. In humans with AMD, MCs accumulated in the subretinal space at sites of retinal degeneration and choroidal neovascularization (CNV). In CX3CR1-deficient mice, MCs accumulated subretinally with age and albino background and after laser impact preceding retinal degeneration. Raising the albino mice in the dark prevented both events. The appearance of lipid-bloated subretinal MCs was drusen like on funduscopy of senescent mice, and CX3CR1-dependent MC accumulation was associated with an exacerbation of experimental CNV. These results show that CX3CR1-dependent accumulation of subretinal MCs evokes cardinal features of AMD. These findings reveal what we believe to be a novel pathogenic process with important implications for the development of new therapies for AMD. PMID- 17909629 TI - Pten controls lung morphogenesis, bronchioalveolar stem cells, and onset of lung adenocarcinomas in mice. AB - PTEN is a tumor suppressor gene mutated in many human cancers. We generated a bronchioalveolar epithelium-specific null mutation of Pten in mice [SP-C rtTA/(tetO)(7)-Cre/Pten(flox/flox) (SOPten(flox/flox)) mice] that was under the control of doxycycline. Ninety percent of SOPten(flox/flox) mice that received doxycycline in utero [SOPten(flox/flox)(E10-16) mice] died of hypoxia soon after birth. Surviving SOPten(flox/flox)(E10-16) mice and mice that received doxycycline postnatally [SOPten(flox/flox)(P21-27) mice] developed spontaneous lung adenocarcinomas. Urethane treatment accelerated number and size of lung tumors developing in SOPten(flox/flox) mice of both ages. Histological and biochemical examinations of the lungs of SOPten(flox/flox)(E10-16) mice revealed hyperplasia of bronchioalveolar epithelial cells and myofibroblast precursors, enlarged alveolar epithelial cells, and impaired production of surfactant proteins. Numbers of bronchioalveolar stem cells (BASCs), putative initiators of lung adenocarcinomas, were increased. Lungs of SOPten(flox/flox)(E10-16) mice showed increased expression of Spry2, which inhibits the maturation of alveolar epithelial cells. Levels of Akt, c-Myc, Bcl-2, and Shh were also elevated in SOPten(flox/flox)(E10-16) and SOPten(flox/flox)(P21-27) lungs. Furthermore, K-ras was frequently mutated in adenocarcinomas observed in SOPten(flox/flox)(P21-27) lungs. These results indicate that Pten is essential for both normal lung morphogenesis and the prevention of lung carcinogenesis, possibly because this tumor suppressor is required for BASC homeostasis. PMID- 17909630 TI - Macrophages suppress T cell responses and arthritis development in mice by producing reactive oxygen species. AB - Reduced capacity to produce ROS increases the severity of T cell-dependent arthritis in both mice and rats with polymorphisms in neutrophil cytosolic factor 1 (Ncf1) (p47phox). Since T cells cannot exert oxidative burst, we hypothesized that T cell responsiveness is downregulated by ROS produced by APCs. Macrophages have the highest burst capacity among APCs, so to study the effect of macrophage ROS on T cell activation, we developed transgenic mice expressing functional Ncf1 restricted to macrophages. Macrophage-restricted expression of functional Ncf1 restored arthritis resistance to the level of that of wild-type mice in a collagen-induced arthritis model but not in a T cell-independent anti-collagen antibody-induced arthritis model. T cell activation was downregulated and skewed toward Th2 in transgenic mice. In vitro, IL-2 production and T cell proliferation were suppressed by macrophage ROS, irrespective of T cell origin. IFN-gamma production, however, was independent of macrophage ROS but dependent on T cell origin. These effects were antigen dependent but not restricted to collagen type II. In conclusion, macrophage-derived ROS play a role in T cell selection, maturation, and differentiation, and also a suppressive role in T cell activation, and thereby mediate protection against autoimmune diseases like arthritis. PMID- 17909633 TI - Science education and communication: AAP Presidential Address. PMID- 17909634 TI - Introduction of Anthony S. Fauci, MD: 2007 Association of American Physicians George M. Kober Medal. PMID- 17909631 TI - HLA class I polymorphisms are associated with development of infectious mononucleosis upon primary EBV infection. AB - Infectious mononucleosis (IM) is an immunopathological disease caused by EBV that occurs in young adults and is a risk factor for Hodgkin lymphoma (HL). An association between EBV-positive HL and genetic markers in the HLA class I locus has been identified, indicating that genetic differences in the HLA class I locus may alter disease phenotypes associated with EBV infection. To further determine whether HLA class I alleles may affect development of EBV-associated diseases, we analyzed 2 microsatellite markers and 2 SNPs located near the HLA class I locus in patients with acute IM and in asymptomatic EBV-seropositive and -seronegative individuals. Alleles of both microsatellite markers were significantly associated with development of IM. Specific alleles of the 2 SNPs were also significantly more frequent in patients with IM than in EBV-seronegative individuals. IM patients possessing the associated microsatellite allele had fewer lymphocytes and increased neutrophils relative to IM patients lacking the allele. These patients also displayed higher EBV titers and milder IM symptoms. The results of this study indicate that HLA class I polymorphisms may predispose patients to development of IM upon primary EBV infection, suggesting that genetic variation in T cell responses can influence the nature of primary EBV infection and the level of viral persistence. PMID- 17909632 TI - Abrogation of macrophage migration inhibitory factor decreases West Nile virus lethality by limiting viral neuroinvasion. AB - The flavivirus West Nile virus (WNV) is an emerging pathogen that causes life threatening encephalitis in susceptible individuals. We investigated the role of the proinflammatory cytokine macrophage migration inhibitory factor (MIF), which is an upstream mediator of innate immunity, in WNV immunopathogenesis. We found that patients suffering from acute WNV infection presented with increased MIF levels in plasma and in cerebrospinal fluid. MIF expression also was induced in WNV-infected mice. Remarkably, abrogation of MIF action by 3 distinct approaches (antibody blockade, small molecule pharmacologic inhibition, and genetic deletion) rendered mice more resistant to WNV lethality. Mif(-/-) mice showed a reduced viral load and inflammatory response in the brain when compared with wild type mice. Our results also indicate that MIF favors viral neuroinvasion by compromising the integrity of the blood-brain barrier. In conclusion, the data obtained from this study provide direct evidence for the involvement of MIF in viral pathogenesis and suggest that pharmacotherapeutic approaches targeting MIF may hold promise for the treatment of WNV encephalitis. PMID- 17909635 TI - A view from Washington through the eyes of an AAP physician-scientist: 2007 Association of American Physicians George M. Kober Medal. PMID- 17909637 TI - Nanotechnology applications and implications research supported by the US Environmental Protection Agency STAR grants program. AB - Since 2002, the US Environmental Protection Agency (EPA) has been funding research on the environmental aspects of nanotechnology through its Science to Achieve Results (STAR) grants program. In total, more than $25 million has been awarded for 86 research projects on the environmental applications and implications of nanotechnology. In the applications area, grantees have produced promising results in green manufacturing, remediation, sensors, and treatment using nanotechnology and nanomaterials. Although there are many potential benefits of nanotechnology, there has also been increasing concern about the environmental and health effects of nanomaterials, and there are significant gaps in the data needed to address these concerns. Research performed by STAR grantees is beginning to address these needs. PMID- 17909638 TI - The environmental behaviour of polychlorinated phenols and its relevance to cork forest ecosystems: a review. AB - Pentachlorophenol (PCP) has been used as a herbicide, biocide and preservative worldwide since the 1930s and as a result, extensive and prolonged contamination exists. The environmental impact increases when its many degradation products are taken into consideration. A number of chloroanisols and their related chlorophenols have been found in cork slabs collected from Portuguese oak tree forests before stopper manufacturing, and contamination by PCP and polychlorinated anisole (PCA) has been detected in Canadian forests. It is suggested that the use of polychlorinated phenols, in particular PCP, is thought to be a cause of the cork taint problem in wine, a major socio-economic impact not only for industry but on sensitive and highly biodiverse ecosystems. It also highlights particular issues relating to the regional regulation of potentially toxic chemicals and global economics world wide. To fully understand the impact of contamination sources, the mechanisms responsible for the fate and transport of PCP and its degradation products and assessment of their environmental behaviour is required. This review looks at the current state of knowledge of soil sorption, fate and bioavailability and identifies the challenges of degradation product identification and the contradictory evidence from field and laboratory observations. The need for a systematic evaluation of PCP contamination in relation to cork forest ecosystems and transfer of PCP between trophic levels is emphasised by discrepancies in bioaccumulation and toxicity. This is essential to enable long term management of not only transboundary contaminants, but also the sustainable management of socially and economically important forest ecosystems. PMID- 17909639 TI - The combination of oxalic acid with power ultrasound fully degrades chrysotile asbestos fibres. AB - The simultaneous action of power ultrasound and oxalic acid, as a chelating agent, rapidly converts chrysotile asbestos into water soluble material and a non asbestos debris, not classifiable as hazardous under worldwide safety regulations. PMID- 17909640 TI - Additional danger of arsenic exposure through inhalation from burning of cow dung cakes laced with arsenic as a fuel in arsenic affected villages in Ganga-Meghna Brahmaputra plain. AB - In arsenic contaminated areas of the Ganga-Meghna-Brahmaputra (GMB) plain (area 569,749 sq. km; population over 500 million) where traditionally cow dung cake is used as a fuel in unventilated ovens for cooking purposes, people are simply exposed to 1859.2 ng arsenic per day through direct inhalation, of which 464.8 ng could be absorbed in respiratory tract. PMID- 17909641 TI - PM sources in a highly industrialised area in the process of implementing PM abatement technology. Quantification and evolution. AB - Principal component analysis (PCA) coupled with a multilinear regression analysis (MLRA) was applied to PM(10) speciation data series (2002-2005) from four sampling sites in a highly industrialised area (ceramic production) in the process of implementing emission abatement technology. Five common factors with similar chemical profiles were identified at all the sites: mineral, regional background (influenced by the industrial estate located on the coast: an oil refinery and a power plant), sea spray, industrial 1 (manufacture and use of glaze components, including frit fusion) and road traffic. The contribution of the regional background differs slightly from site to site. The mineral factor, attributed to the sum of several sources (mainly the ceramic industry, but also with minor contributions from soil resuspension and African dust outbreaks) contributes between 9 and 11 microg m(-3) at all the sites. Source industrial 1 entails an increase in PM(10) levels between 4 and 5 microg m(-3) at the urban sites and 2 microg m(-3) at the suburban background site. However, after 2004, this source contributed less than 2 microg m(-3) at most sites, whereas the remaining sources did not show an upward or downward trend along the study period. This gradual decrease in the contribution of source industrial 1 coincides with the implementation of PM abatement technology in the frit fusion kilns of the area. This relationship enables us to assess the efficiency of the implementation of environmental technologies in terms of their impact on air quality. PMID- 17909642 TI - Atmospheric transport of persistent organic pollutants (POPs) to Bjornoya (Bear island). AB - A first medium term monitoring of atmospheric transport and distribution for persistent organic pollutants (POPs) in Bjornoya (Bear island) air samples has been performed in the period between week 51/1999 and week 28/2003. A total of 50 single compounds consisting of polychlorinated biphenyls (33 congeners), hexachlorobenzene (HCB), hexachlorocyclohexane isomers (alpha-, beta-, gamma HCH), alpha-endosulfan, cyclodiene pesticides (chlordanes, nonachlor-isomers, oxy chlordane, heptachlor and chlordane) as well as dichlorodiphenyltrichloroethane (DDT) derivatives were analysed and quantified. Atmospheric transport of POPs was identified as an important contamination source for the island. PCBs, HCB and HCH isomers were the predominant POP groups, contributing with 70-90% to the overall POP burden quantified in the Bjornoya air samples. The highest concentration levels for a single compound were found for HCB (25-35 pg m(-3)). However, the sum of 33 PCB congeners was found to be in the same concentration range (annual means between 15 and 30 pg m(-3)). Cyclodiene pesticides, DDT derivatives and alpha-endosulfan were identified as minor contaminants. Several atmospheric long range transport episodes were identified and characterised. Indications for industrial emissions as well as agricultural sources were found for the respective atmospheric transport episodes. A first simple statistical correlation assessment showed that for long-range transport of pollution, the local meteorological situation is not as important as the air mass properties integrated over the time period of the transport event. The local weather situation, on the other hand, is important when investigating deposition rates and up-take/accumulation properties in the local ecosystem. Based upon chemical data interpretation, valuable information about the influence of primary and secondary sources on the air mass contamination with chlorinated insecticides (e.g., HCHs) was found and discussed. The interdisciplinary interpretation of contaminant data using statistical methods, chemical analysis, meteorological modelling and classical meteorological information for a comprehensive evaluation of atmospheric long range transport into the European Arctic (Bjornoya) has proven to be a highly versatile tool not only for atmospheric scientists but also with strong potential for regulatory purposes. PMID- 17909643 TI - Polycyclic aromatic hydrocarbons (PAHs) in the air of Chinese cities. AB - Polycyclic aromatic hydrocarbons (PAHs) were determined in the air of 37 cities and 3 rural locations across China during the winter, spring, summer and autumn of 2005, using polyurethane foam (PUF) disks as passive air samplers (PAS). Winter and autumn concentrations in cities exceeded spring and summer values. Concentrations were amongst the highest in the world; seasonally averaged autumn/winter values in some cities in the north and north-west of China exceeded proposed European Union air quality standards. Several factors, acting in combination, influenced air concentrations. A significant negative correlation was found between average annual city concentrations and the annual average temperature, while winter time PAH concentrations correlated with estimated coal consumption. The highest total PAH concentrations and loadings of high molecular weight compounds generally occurred in major cities located on higher land (500 2000 m), where relatively cold winters and higher coal consumption occurs. Lower values occurred in cities located in the south and east China and along the coastal regions. Molecular markers indicated incomplete combustion of fossil fuels dominated the urban air and gave evidence for photo-decomposition of selected compounds. PMID- 17909644 TI - An inexpensive light-scattering particle monitor: field validation. AB - We have developed a small, light, passive, inexpensive, datalogging particle monitor called the "UCB" (University of California Berkeley particle monitor). Following previously published laboratory assessments, we present here results of tests of its performance in field settings at high particle concentrations. We demonstrate the mass sensitivity of the UCB in relation to gravimetric filter based PM(2.5) mass estimates as well as commercial light-scattering instruments co-located in field chamber tests and in kitchens of wood-burning households. The coefficient of variation of the unadjusted UCB mass response in relation to gravimetric estimates was 15%. Although requiring adjustment for differences in sensitivity, inter-monitor performance was consistently high (r(2) > 0.99). Moreover, the UCB can consistently estimate PM(2.5) mass concentrations in wood burning kitchens (Pearson r(2) = 0.89; N = 99), with good agreement between duplicate measures (Pearson r(2) = 0.94; N = 88). In addition, with appropriate cleaning of the sensing chamber, UCB mass sensitivity does not decrease with time when used intensively in open woodfire kitchens, demonstrating the significant potential of this monitor. PMID- 17909645 TI - Grain-size analysis of volcanic ash for the rapid assessment of respiratory health hazard. AB - Volcanic ash has the potential to cause acute and chronic respiratory diseases if the particles are sufficiently fine to enter the respiratory system. Characterization of the grain-size distribution (GSD) of volcanic ash is, therefore, a critical first step in assessing its health hazard. Quantification of health-relevant size fractions is challenging without state-of-the-art technology, such as the laser diffractometer. Here, several methods for GSD characterization for health assessment are considered, the potential for low-cost measurements is investigated and the first database of health-pertinent GSD data is presented for a suite of ash samples from around the world. Methodologies for accurate measurement of the GSD of volcanic ash by laser diffraction are presented by experimental analysis of optimal refractive indices for different magmatic compositions. Techniques for representative sampling of small quantities of ash are also experimentally investigated. GSD results for health-pertinent fractions for a suite of 63 ash samples show that the fraction of respirable (<4 microm) material ranges from 0-17 vol%, with the variation reflecting factors such as the style of the eruption and the distance from the source. A strong correlation between the amount of <4 and <10 microm material is observed for all ash types. This relationship is stable at all distances from the volcano and with all eruption styles and can be applied to volcanic plume and ash fallout models. A weaker relationship between the <4 and <63 microm fractions provides a novel means of estimating the quantity of respirable material from data obtained by sieving. PMID- 17909646 TI - Passive sampling: partition coefficients for a silicone rubber reference phase. AB - Silicone rubber sheeting can be used as a passive sampling device for hydrophobic organic contaminants in the environment to determine the available concentrations in water and sediments. Reliable sampler-water partition coefficients are required to determine the sampling rates and the dissolved contaminant concentrations in water and in sediment pore water. Log partition coefficients (logK(sr,w)) for silicone rubber-water have been estimated for 32 polycyclic aromatic hydrocarbons (PAHs), 2 deuterated PAH analogues and 32 chlorobiphenyls (CBs) using the cosolvent method, with methanol as cosolvent. Strong linear relationships were found with literature values for the corresponding log octanol water partition coefficients (logK(ow)) for both CBs and PAHs, confirming that partitioning into the silicone rubber is strongly determined by the hydrophobicity of the compounds, which suggests logK(ow) is a good predictor of logK(sr,w) and that absorption is the main mechanism for accumulation of analytes into the silicone rubber polymer. PMID- 17909647 TI - Investigation of reagent distributions on glass fiber membrane filters used in air sampling. AB - This project has arisen from the need to produce GFFs (glass fiber filters) bearing a thin and evenly distributed coating of a selected reagent in the equatorial plane for breakthrough studies. However, it has been discovered that today's two general techniques for coating GFFs (total immersion and application of reagent solution to GFFs) have usually produced unevenly distributed coatings of reagent in the equatorial plane. In addition, quantities of reagent on GFFs from commercial sources may vary widely in the same lot of coated GFFs. Consequences are variability in capacity of coated filters at the point of breakthrough and, perhaps, wasted reagent. Although today's reagent-coated filters may be satisfactory for routine air sampling, such filters may be unacceptable for precise breakthrough studies. Research has been conducted successfully to produce nearly evenly distributed coatings of reagents in the equatorial plane of GFFs by application of reagent solutions to the centers of GFFs which are resting on crisscrossing, fine, stainless-steel wire. Distributions of coatings have been determined by punching out twenty-one 5-mm circles from each GFF and analyzing each circle by flow-injection with a UV detector. Lowest achievable relative standard deviations of measurement (RSDs) for reagents in 5-mm circles have been 5 to 7%. Reagents studied have included 1 (2-pyridyl)piperazine (1-2PP), 2,4-dinitrophenylhydrazine (DNPH), and 1-(9 anthracenylmethyl)piperazine (MAP). Factors affecting the distribution of such coatings include choice of reagent and choice of solvent for the reagent solution. PMID- 17909648 TI - Insights into porphyrin chemistry provided by the microperoxidases, the haempeptides derived from cytochrome c. AB - The water-soluble haem-containing peptides obtained by proteolytic digestion of cytochrome c, the microperoxidases, have been used to explore aspects of the chemistry of iron porphyrins, and as mimics for some reactions catalysed by the haemoproteins, including the reactions catalysed by the peroxidases and the cytochromes P450. The preparation of the microperoxidases, their physical and chemical properties including their electronic structure, the kinetics and thermodynamics of their reactions with ligands, electrochemical studies and examples of their uses as haemoproteins mimics, is reviewed. PMID- 17909649 TI - The first example of a Pt...Pt interaction in platinum(ii) complexes bearing bulky tri-tert-butyl-2,2':6',2''-terpyridine pendants via conformational control of the calix[4]arene moiety. AB - An unprecedented short Pt...Pt contact between sterically bulky Pt((t)Bu(3)trpy) alkynyl moieties has been observed in the X-ray crystal structure of a dinuclear platinum(ii) complex bridged by a diethynylcalix[4]arene derivative; the complex in its crystalline state showed a red shift in the emission maxima at 298 K and 77 K relative to its powder form, which has been attributed to the presence of a metal-metal interaction in the crystal lattice. PMID- 17909650 TI - Neutral bis(1,4-diaza-1,3-butadiene)nickel complexes and their corresponding monocations: molecular and electronic structures. A combined experimental and density functional theoretical study. AB - The reaction of 2 equivalents of 2-methyl-1,4-bis(2,6-dimethylphenyl)-1,4-diaza 1,3-butadiene, ((1)L(Ox))(0), or 2-methyl-1,4-bis(2,6-diisopropylphenyl)-1,4 diaza-1,3-butadiene, ((2)L(Ox))(0), in diethyl ether or n-hexane with 1 equivalent of Ni(cdt) where (cdt)(0) is the ligand cyclododecatriene affords dark red, diamagnetic precipitates of [Ni(II)((1)L )(2)] () and of [Ni(II)((2)L )(2)] (). The ligands ((1)L )(1-) and ((2)L )(1-) are the one-electron reduced, monoanionic pi radicals of the above neutral 1,4-diaza-1,3-butadienes. and have been structurally characterized by X-ray crystallography; both possess a distorted tetrahedral geometry where the dihedral angle theta between the two metallacycles Ni-N-C-C-N is 47.9 degrees and 53 degrees , respectively, (theta = 0 degrees for square planar and 90 degrees for a regular tetrahedral geometry). The reaction of and with 1 equivalent of ferrocenium hexafluorophosphate gives the paramagnetic (S(t) = 1/2) complexes and , respectively: [Ni(I)((1)L(Ox))(2)](PF(6)) (), [Ni(I)((2)L(Ox))(2)](PF(6)) (). Their EPR spectra indicate the presence of a central Ni(i) ion (d(9); S(Ni) = 1/2). Thus, the one electron oxidation of and by [Fc]PF(6) induces an intramolecular one-electron reduction of the central Ni ion and a concomitant one-electron oxidation of the second pi radical (L )(1-)--> (L(Ox))(0) + e. Broken symmetry DFT calculations (B3LYP) corroborate the correctness of the electronic structure descriptions of . The reaction of ((1)L(Ox))(0) with NiI(2) (1 : 1) in tetrahydrofuran yields tetrahedral [Ni(II)((1)L(Ox))I(2)] () with an S(t) = 1 ground state. PMID- 17909652 TI - Insertion reactions of dicyclohexylcarbodiimide with aminoboranes, -boryls and borylenes. AB - Insertion reactions of dicyclohexylcarbodiimide with aminoboranes and with aminoboryl and -borylene transition metal complexes have been examined as potential routes to new boron-containing ligand systems. Reactions with systems containing two-coordinate boron centres are found to be significantly more facile than those with three-coordinate substrates. Thus, reaction of (dicyclohexylamino)boron dichloride () with dicyclohexylcarbodiimide over 36 h at 50 degrees C generates the (structurally authenticated) guanidinate complex Cy(2)NC(NCy)(2)BCl(2) () via insertion into the BN bond. By contrast, the corresponding reaction with the cationic aminoborylene complex [CpFe(CO)(2)(BNCy(2))](+)[BAr(f)(4)](-) () proceeds rapidly at ca.-30 degrees C, via initial insertion into the FeB bond to give [CpFe(CO)(2)C(NCy)(2)BNCy(2)](+)[BAr(f)(4)](-) (). Consistent with related studies, a key factor in facilitating such insertion chemistry is thought to be the formation of an initial donor/acceptor complex between the diimide and the group 13 centre. Thus, DFT studies suggest that [CpFe(CO)(2)B(NCy(2))(CyNCNCy)](+)[BAr(f)(4)](-) is a potential intermediate in the reaction of with CyNCNCy, and that further reaction to give the observed product, , is strongly exergic (-183 kJ mol(-1)). By contrast, DFT calculations for the alternative isomer [CpFe(CO)(2)B(CyN)(2)CNCy(2)](+)[BAr(f)(4)](-) (), formed by BN insertion, suggest that it is 112 kJ mol(-1) less stable than . Such experimental and computational findings imply that under reaction conditions where a suitable isomerisation pathway is available, cationic complexes such as , which contain a four-membered boron-donor heterocycle are likely to be disfavoured with respect to alternative C-bound isomers. PMID- 17909653 TI - Structure, magnetism and photomagnetism of mixed-ligand tris(pyrazolyl)methane iron(ii) spin crossover compounds. AB - A range of bis-facial tridentate chelate complexes of type [Fe((R-pz)(3)CH)((3,5 Me(2)pz)(3)CH)](BF(4))(2) has been characterised that contain two different tris pyrazolylmethane ligands, with variations in R being H (complex crystallised as polymorphs and ) and 4-Me (), as well as R = H with a CH(2)OH arm off the methane carbon (). A tris(pyridyl)methane analogue is also described (). The tris(3,5 dimethylpyrazolyl)methane co-ligand (3,5-Me(2)pz), and the BF(4)(-) counterion, are constant throughout. The spin-crossover properties of these Fe(ii) d(6) compounds have been probed in detail by variable temperature magnetic, Mossbauer spectral and crystallographic methods. The effects of distortions from octahedral symmetry around the Fe(ii) centres, of crystal solvate molecules (1.5 MeCN in and 2 MeCN in ) and of supramolecular/crystal packing, are discussed. In the case of , subtle twisting of pyrazole rings occurs, as a function of temperature, that has a greater effect upon the relative positions of the Fe(ii) chelate molecules in polymorph than in polymorph ; this is thought to drive the cooperativity differences observed in the magnetism of the polymorphs. Comparisons are also made between to and their homoleptic, parent [Fe(L)(2)] (2+) materials. The complexes were screened for the LIESST (light induced excited spin state trapping) effect by measurements of diffuse absorption spectra on the surface of powder samples, at different temperatures. One example, , showed a 2-step thermal spin crossover transition and it was probed in detail for its photomagnetic features. The T(LIESST) and T(1/2) values for did not obey an empirical relationship, T(LIESST) = 150 - 0.3T(1/2) followed by many Fe(ii)(N-donor)(6) crossover compounds of the bis-tridentate (meridional) type, and possible reasons for this are discussed. PMID- 17909654 TI - Transition metal vinylidene complexes as supramolecular building blocks: nucleobase-mediated self-assembly of crystals with hexagonal symmetry. AB - Reaction of the ruthenium half sandwich compound RuCl(eta(5)-C(5)H(5))(PPh(3))(2) with the uracil (Ur) substituted alkyne HC[triple bond, length as m-dash]CUr in the presence of halide scavengers NH(4)X (X = PF(6), BF(4), OTf) results in the formation of the vinylidene complexes [Ru([double bond, length as m-dash]C[double bond, length as m-dash]CHUr)(eta(5)-C(5)H(5))(PPh(3))(2)][X] which crystallize in the hexagonal space group P6(3)/m. The hexagonal symmetry inherent to the system is due to the formation of a hydrogen bonded array mediated by the two sets of donor-acceptor units on the uracil, resulting in the formation of a cyclic "rosette" containing six ruthenium cations. In solution the (1)H and (31)P{(1)H} NMR spectra of the vinylidene complexes are both concentration and temperature dependent, in accord with the presence of monomer-dimer equilibria in which the rate of rotation of the vinylidene group is fast on the NMR timescale in the monomeric species, but slow in the dimers. The isoelectronic molybdenum containing vinylidene complex [Mo(eta(7)-C(7)H(7))(dppe)([double bond, length as m-dash]C[double bond, length as m-dash]CHUr)][BF(4)] (dppe = 1,2 bis(diphenylphosphino)ethane) has also been prepared, but forms symmetric dimers in the solid state. PMID- 17909651 TI - Dinuclear palladium(ii) complexes with bridging amidate ligands. AB - New homonuclear dimeric Pd(ii) complexes have been synthesized by the reaction of Pd(en)(2+) or Pd(bipy)(2+) (where en = ethylenediamine and bipy = 2,2' bipyridine) units with acetamide or by the Pd(ii) mediated hydrolysis of CH(3)CN. In these dimers the two metal centers are bridged by either two amidates or by the combination of one hydroxo group and one amidate ligand. The crystal structures of complexes {[Pd(bipy)](2)(micro-1,3 CH(3)CONH)(2)}(NO(3))(2).H(2)O.1/2(CH(3))(2)CO.1/2CH(3)CN () and {[Pd(bipy)](2)(micro-1,3-CH(3)CONH)(2)}(OTf)(2) () showed intrametallic Pd-Pd distances of 2.8480(8) A () and 2.8384(7) A (), respectively, in accordance with the accepted values for a strong Pd-Pd interaction. The presence of pi[dot dot dot]pi interactions between the bipyridine ligands on the di-micro-amidate complexes of Pd(bipy)(2+) shortens the distance between the two Pd centers and allows the formation of the metal-metal interaction. By contrast, the crystal structure of complex {[Pd(en)](2)(micro-1,3-CH(3)CONH)(2)}(OTf)(2).H(2)O (), (where OTf = triflate) where there is no pi[dot dot dot]pi interaction between the ligands on the metal centers, is also reported, and no Pd-Pd interaction is observed. Additionally, one of the complexes, {[Pd(en)](2)(micro-OH)(micro CH(3)CONH)}(NO(3))(2) (), presents an interesting hydrogen bonded 3-D network formed by nitrate ions and water molecules. All complexes have been characterized by infrared and (1)H NMR spectroscopy. PMID- 17909655 TI - One-pot syntheses, coordination, and characterization of application-specific biodegradable ligand-polymers. AB - Syntheses and chelation of tailored biodegradable polymers to rhenium for medicinal applications are described. A group of bifunctional ligand-initiators consisting of a chelating end for metal complexation and a hydroxyl end suitable to initiate polymerization was utilized in the ring-opening polymerization of l lactide. The resulting biodegradable ligand-polymers were equipped with a tridentate donor set to coordinate specific metal ions. All synthesized compounds were characterized by IR spectroscopy, 1D/2D NMR spectroscopy and MALDI-TOF mass spectrometry, confirming successful polymerization and coordination to the [Re(CO)(3)](+) core. The pliability of designing application-specific polymers with respect to the nature of the metal ion facilitates extending the application of these biodegradable polymers to early detection of diseases (imaging) and radiotherapy of cancers. PMID- 17909656 TI - Synthesis, structures and properties of a new series of platinum-diimine dithiolate complexes. AB - The new square-planar platinum-diimine-dithiolate compounds [Pt(mesBIAN)SS] have been synthesised {mesBIAN = bis(mesityl)biazanaphthenequinone; SS = 1,2 dithiooxalate (dto) , maleonitriledithiolate (mnt) , 1,2-benzenedithiolate (bdt) , 3,4-toluenedithiolate (tdt) and 1,3-dithia-2-thione-4,5-dithiolate (dmit) }, and the X-ray crystal structures of and determined. Cyclic voltammetry reveals that all the compounds form stable anions, and ESR spectroscopy of these anions shows that the SOMO is based upon the mesBIAN ligand; compounds also show a reversible oxidation wave in their CV. Computational studies reveal that charge transfer processes from orbitals that are combinations of metal and dithiolate ligand to a mesBIAN pi-based LUMO are responsible for the low energy absorptions seen in the UV/visible spectra of these compounds, and that the reverse process is responsible for the observed room-temperature solution luminescence of [Pt(mesBIAN)Cl(2)] and , and . Compounds and , containing aromatic thiolates, were not found to luminesce under the same conditions. Resonance Raman experiments have shown the origin of band-broadening of the lowest-energy absorption band in the absorption spectra of to be due to vibronic structure within one electronic transition. PMID- 17909657 TI - Chiral or not chiral? A case study of the hexanuclear metalloprisms [Cp(6)M(6)(micro(3)-tpt-kappaN)(2)(micro-C2O4-kappaO)(3)]6+ (M = Rh, Ir, tpt = 2,4,6-tri(pyridin-4-yl)-1,3,5-triazine). AB - Cationic hexarhodium and hexairidium complexes with a trigonal prismatic architecture have been synthesised in good yield by self-assembly of the dinuclear oxalato-bridged complexes [Cp(2)M(2)(micro-C(2)O(4)-kappaO)Cl(2)] (M = Rh; 1: Ir; 2) with 2,4,6-tri(pyridine-4-yl)-1,3,5-triazine (tpt) in the presence of AgO(3)SCF(3). The trigonal prismatic cations [Cp(6)Rh(6)(micro(3)-tpt kappaN)(2)(micro-C(2)O(4)-kappaO)(3)](6+) (3) and [Cp(6)Ir(6)(micro(3)-tpt kappaN)(2)(micro-C(2)O(4)-kappaO)(3)](6+) (4) have been isolated as their triflate salts. The single-crystal X-ray structure analysis of [3][O(3)SCF(3)](6) shows two enantiomers in the racemic crystal (space group C2/c), the chirality being due to a twist of the two tpt units. By contrast, the single-crystal X-ray structure analysis of [4][O(3)SCF(3)](6) shows a perfectly eclipsed conformation of the tpt units, so that is not chiral in the crystal state (space group Fd3[combining macron]c). However, in solution, enantiodifferentiation in the presence of the chiral anion Delta-BINPHAT is observed by (1)H NMR spectrometry not only in the case of 3, but also in the case of 4. This suggests that the iridium derivative 4, which is not chiral in the solid state, adopts chiral conformations in solution. PMID- 17909658 TI - Tert-butylamidinate tin(ii) complexes: high activity, single-site initiators for the controlled production of polylactide. AB - The tin(ii) coordination chemistry of two monoanionic N,N'-bis(2,6 diisopropylphenyl)alkylamidinate ligands is described. Complexation studies with the acetamidinate, [MeC(NAr)(2)](-), (Ar = 2,6-(i)Pr(2)C(6)H(3)) are complicated by the side formation of the bis(amidinate) tin(ii) compound, [MeC(NAr)(2)](2)Sn. By contrast, the bulkier tert-butylamidinate, [(t)BuC(NAr)(2)](-), allows tin(ii) mono-halide, -alkoxide and -amide complexes to be isolated cleanly in high yields. Thus, the reaction of [(t)BuC(NAr)(2)]H with (n)BuLi and subsequent treatment with SnCl(2) generates [(t)BuC(NAr)(2)]SnCl, in ca. 70% yield. Reactions of with LiO(i)Pr, LiNMe(2) and LiNTMS(2) afford [(t)BuC(NAr)(2)]Sn(O(i)Pr), [(t)BuC(NAr)(2)]Sn(NMe(2)), and [(t)BuC(NAr)(2)]Sn(NTMS(2)), respectively. The molecular structures of complexes are reported. Complexes, and have been investigated as initiators for the ring opening polymerisation of rac-lactide: and display characteristics of well controlled polymerisation initiators, but high molecular weight polymer is observed with due to inefficient initiation, a consequence of the steric bulk of the NTMS(2) unit. Polymerisations with and are faster than for the corresponding beta-diketiminate tin(ii) complexes, consistent with the more open nature of the tin(ii) coordination sphere. PMID- 17909659 TI - Mono- and dinuclear cobalt complexes with chelating or bridging bidentate P,N phosphino- and phosphinito-oxazoline ligands: synthesis, structures and catalytic ethylene oligomerisation. AB - Cobalt(ii) complexes of the type [CoCl(2)(P,N)], where P,N represents a heterobidentate phosphino- or phosphinito-oxazoline-type ligand, have been synthesised and characterised by infrared spectroscopy and elemental analysis. Their molecular structures were established by single-crystal X-ray diffraction in the solid state. Whereas the phosphino-oxazoline complex [CoCl(2){Ph(2)PCH(2)ox(Me2)}] (Ph(2)PCH(2)ox(Me2) = 2-[(diphenylphosphanyl) methyl]-4,4-dimethyl-4,5-dihydro-oxazole) () and the phosphinito-oxazoline complexes [CoCl(2){Ph(2)POCH(2)ox(Me2)}] (Ph(2)POCH(2)ox(Me2) = 1-[4,4-dimethyl 2{1-oxy(diphenylphosphino)-1-methyl}]-4,5-dihydro-oxazole) () and [CoCl(2){Ph(2)POCMe(2)ox(Me2)}] (Ph(2)POCMe(2)ox(Me2) = 1-[4,4-dimethyl-2- [1 oxy(diphenylphosphino)-1-methylethyl]]-4,5-dihydrooxazole) () are mononuclear, the phosphino-oxazoline complexes [CoCl(2){micro-i-Pr(2)PCH(2)ox}](2) (i Pr(2)PCH(2)ox = 2-[(diisopropyl-phosphanyl)-methyl]-4,5-dihydro-oxazole) () and [CoCl(2){micro-Ph(2)PCH(2)ox}](2) (Ph(2)PCH(2)ox = 2-[(diphenyl-phosphanyl) methyl]-4,5-dihydro-oxazole) () are dinuclear compounds and contain two bridging phosphino-oxazoline ligands which form a 10-membered ring. In the course of this work, the zwitterionic complex [CoCl(3){Ph(2)PCH(2)C(O)OCH(2)CMe(2)NH(3)] () was obtained and characterised by X-ray diffraction in which the oxazoline ring has been opened. Air-oxidation of the phosphine function of the mononuclear P,N chelate complex yielded the blue N,O-bridged, centrosymmetric dinuclear complex [[upper bond 1 start]CoCl(2){micro-OPPh(2)CH(2)[lower bond 1 start]C[double bond, length as m-dash]N[upper bond 1 end]CMe(2)CH(2)O[lower bond 1 end]}](2) () which contains a 12-membered ring. All these complexes are paramagnetic and their magnetic moments in solution were measured by the Evans method. Complexes were evaluated in the catalytic oligomerisation of ethylene with AlEtCl(2) or methylaluminoxane (MAO) as cocatalysts and provided moderate activities. In the presence of AlEtCl(2) (6-14 equiv.), the selectivities for ethylene dimers were higher than 92% and complex showed the highest turnover frequency with 14 equiv. of AlEtCl(2). When MAO was used as cocatalyst, the catalytic activities were similar to those with AlEtCl(2) but significant amounts of C(6)-C(12) oligomers were produced. PMID- 17909660 TI - Coronary artery fistula diagnosed by transthoracic Doppler echocardiography. AB - A 47-year-old Malay woman complained of an episode of shortness of breath after a shower. There was no previous complaint of shortness of breath or chest pain. Physical examination revealed a wide pulse pressure. Blood pressure was 160/66 mmHg, and heart rate was 77/minute and regular. What was initially thought to be a loud pansystolic murmur was heard over the precordium. Electrocardiography showed left ventricular hypertrophy with a volume overload pattern. Transthoracic Doppler echocardiography revealed a right coronary artery-right ventricular fistula, arising from the right coronary artery and draining into the right ventricular cavity. PMID- 17909661 TI - Cystic lymphangioma of the mesentery causing intestinal obstruction. AB - Mesenteric cystic lymphangioma is a rare lesion that is not often described in the literature. A four-year-old boy, who presented with abdominal distension and pain, is reported. At surgery, a huge mesenteric cyst was found to be the cause of the intestinal obstruction and was completely excised. Histology was consistent with a cystic lymphangioma. Abdominal lymphangioma is a rare cause of bowel obstruction. Clinical presentation varies and may be misleading due to a lack of awareness of the clinical condition. Occasionally, the diagnosis is made during surgery. General awareness of this entity with a high index of suspicion is needed to avoid complications. PMID- 17909662 TI - Cirsoid aneurysm of the scalp. AB - A 29-year-old man with a pulsatile scalp swelling is presented. The clinical diagnosis of a cirsoid aneurysm was confirmed on computed tomography of the brain and selective cerebral angiography. It is important to detect veins draining from the aneurysm into the intracerebral venous system, as percutaneous occlusion of the aneurysm may be possible in their absence. PMID- 17909663 TI - Traumatic abdominal wall herniation. AB - Most reported cases of traumatic abdominal wall herniation result from seatbelt or handlebar injuries. The diagnosis is often made on physical examination or abdominal computed tomography (CT). We report a 59-year-old man with traumatic herniation through the rectus muscle following low-velocity blunt abdominal trauma. This patient was initially thought to have a rectus sheath haematoma and initial CT showed a soft tissue haematoma over the left lower anterior abdominal wall but no herniation. The traumatic herniation was diagnosed four days later, and confirmed on CT. Intraoperatively, a segment of the sigmoid colon was found to have herniated through the rectus defect and was gangrenous with impending perforation. A left hemicolectomy followed by primary repair of the defect was done. This case highlights the need for a high index of suspicion for traumatic herniation in patients who sustain low-velocity blunt abdominal wall trauma even when initial CT scans are negative. PMID- 17909665 TI - Leiomyoma of the nose. AB - Leiomyomas are benign neoplasms that are thought to originate from the vascular smooth muscle. They have a propensity to arise from the gastrointestinal tract, female genital tract (uterus) and subcutaneous tissue. The nasal cavity is an uncommon site for a leiomyoma. We report a 24-year-old woman with a rare nasal leiomyoma. A brief review of the literature and histological variations are described. PMID- 17909664 TI - Mucoepidermoid carcinoma of the tongue. AB - We report a 27-year-old Congolese man with mucoepidermoid carcinoma (MEC) of the tongue base, which presented as spontaneous intraoral bleeding. Optimal treatment of tongue base MEC is unknown. To our knowledge, this is the first reported case treated with transoral excision with carbon dioxide laser and selective neck dissection. Although immunohistochemical studies have revolutionised understanding of the disease, little else is known of the natural history of MEC. The majority of MEC is considered low-grade, with an indolent course without recurrence or metastasis. Nonetheless, MEC requires surgical management, postoperative radiotherapy and close long-term follow-up. PMID- 17909666 TI - Acute vertebrobasilar artery thrombosis: long-term benefit of vertebral artery stenting. AB - Acute vertebrobasilar artery occlusion is a life-threatening event, even after thrombolytic treatment with local intraarterial (IA) recombinant tissue plasminogen activator. We report a 70-year-old man with acute vertebral artery occlusion in which IA thrombolysis resulted in partial recanalisation and revealed pre-existing severe stenosis as the underlying cause. Stenosis was managed with stenting with excellent long-term clinical as well as angiographical outcomes. PMID- 17909667 TI - Dengue haemorrhagic fever complicated by eclampsia in pregnancy. AB - A 28-year-old primigravida presented at 36 weeks of gestation with a one-week history of fever with myalgia. Diagnosis of dengue fever was made based on viral polymerase chain reaction. She progressed to dengue shock syndrome by day nine and subsequently recovered. She delivered a healthy male baby by the vaginal route, but within 24 hours of delivery, had an eclamptic seizure, which was controlled with intravenous magnesium sulphate. Mother and the baby were well at discharge and on the follow-up visit at three months. PMID- 17909668 TI - Singapore nursing in transition: perspectives from the Alice Lee Centre for Nursing Studies, National University of Singapore. PMID- 17909669 TI - Lamotrigine in pregnancy: safety profile and the risk of malformations. AB - The use of antiepileptic drugs in pregnancy always presents challenges to doctors and their patients as it may have deleterious effects on the developing embryo. Lamotrigine is most commonly-prescribed drug among the newer antiepileptic drugs; hence, it has been selected for the present review. A number of studies pertaining to the safety of lamotrigine use during pregnancy have been reported, with differing results. Contradictory results have been reported in animals regarding lamotrigine teratogenicity, and human studies have also proven inconclusive. In many countries, human pregnancy registries are maintained to establish the safety of antiepileptic drugs during pregnancy, as all the different suggestions favour some over others, with specific antiepileptic combinations still being questioned. It is our hope that the present work may integrate the available disparate relevant facts into a directed effort towards minimising the risk of foetal compromise. PMID- 17909670 TI - Bicycle-related injuries: a prospective study of 200 patients. AB - INTRODUCTION: This study aims to evaluate the magnitude, mechanism, distribution and outcome of bicycle-related injuries managed at the Emergency Department, Al Ain Hospital, United Arab Emirates. METHODS: 200 patients, who were treated at the emergency department of Al-Ain Hospital during the period of October 2001 to January 2003, were prospectively studied. A hard copy protocol was designed and data was collected on a daily basis. RESULTS: 175 patients (87.5 percent) were males. The average age was 16.1 +/- 13.7 years. Only two were wearing helmets (one percent). The majority of injuries occurred in the evening and was due to a fall from a bicycle in 163 patients (81.5 percent). 88 patients had lower limb injuries (44 percent), and 72 had head and neck injuries (36 percent). Only 31 patients (15.5 percent) needed hospitalisation. Of these, four (12.9 percent) were admitted to the intensive care unit. The mean (range) hospital stay was 6.3 (1-23) days. Patients who were admitted to the hospital were older males, involved in motor vehicle collisions, and had more head injuries. Three patients (1.5 percent) died. CONCLUSION: Bicyclists' head injuries, caused by a motor vehicle collision, are a main cause of hospital admission. Helmet compliance in our community is alarmingly low, indicating the need for legislation and education on the use of helmets. PMID- 17909671 TI - Subclinical peripheral neuropathy in stable middle-aged patients with chronic obstructive pulmonary disease. AB - INTRODUCTION: Presently, there are few studies addressing the subject of peripheral neuropathy in patients with chronic obstructive pulmonary disease (COPD). Consequently, there is a dearth of evidence and awareness of subclinical neuropathy in stable COPD patients with no significant hypoxaemia, particularly in the age group of 40-60 years. The present study was designed to evaluate the subclinical peripheral neuropathy in this study group. METHODS: 60 subjects were included in the study. The COPD group comprised 30 male smokers with stable COPD, aged between 40 and 60 years and with no clinical neuropathy; and 30 age-matched healthy male volunteers served as the control group. The following nerves were evaluated for latency, amplitude and conduction velocity: for motor nerve conduction - median nerve, ulnar nerve, and common peroneal nerves; and for sensory nerve conduction - median nerve, ulnar nerve, and sural nerves. RESULTS: Five out of 30 COPD patients had peripheral nerve impairment on electrophysiological evaluation. In these patients, we found decreased amplitude and conduction velocity in all examined sensory nerves; however, the conduction velocity was found to be more than 70 percent of the predicted value. These findings were suggestive of predominantly sensory (with milder involvement of motor nerves) axonal polyneuropathy. CONCLUSION: We observed five out of 30 COPD patients to have predominantly sensory axonal peripheral neuropathy. These five COPD patients had significantly higher consumption of cigarettes, longer duration of illness and advanced airflow obstruction when compared to COPD patients with no peripheral neuropathy. PMID- 17909672 TI - Emotional disorders among medical students in a Malaysian private medical school. AB - INTRODUCTION: A study was done between December 2005 and January 2006 to determine the prevalence of emotional disorders among medical students in a private medical school in Ipoh, Perak, Malaysia and to determine the demographical characteristics, contributing factors and the key person consulted for emotional problems. METHODS: Medical students in the private medical school completed the 12-item English version of the General Health Questionnaire (GHQ 12) and a demographical questionnaire. A cut-off point of 3/4 for the GHQ was used to determine negative and positive scores for emotional disorders. RESULTS: Out of 292 medical students, 86.6 percent completed the questionnaires. A total of 117 students (46.2 percent) were found to have emotional disorders. There was no significant association of ethnicity, gender, age group, number of examinations sat, examination performances, past medical conditions and relationships with parents, siblings, course-mates and lecturers with positive GHQ scores. A significant association, however, was found between positive GHQ scores for emotional disorders and the year of study, pressure faced due to examinations, and not having a love relationship. 39 percent of the students stated friends as their main preference for consultation of any emotional problem. CONCLUSION: The prevalence of emotional disorders among medical students was high. Further studies and diagnostic measures are recommended, including a more systematic screening and counselling programme by the medical school for early diagnosis and treatment to prevent complications. PMID- 17909673 TI - Clinical profile and outcome of abdominal tuberculosis in Indian children. AB - INTRODUCTION: Diagnosis of tuberculosis among children poses technical and operational challenges, more so in abdominal tuberculosis (ATB), where the protean clinical manifestations continue to challenge the physicians in its diagnosis and therapy. METHODS: Medical records of 115 patients who were diagnosed with ATB over a period of six years were studied retrospectively. Details of history, physical examination and investigations, treatment and outcome of therapy were evaluated. RESULTS: The mean age of the patients was 6.4 years. Commonest symptom at presentation was abdominal pain, followed by fever. Nine patients presented with acute abdomen. Mantoux test was positive in 33 percent and accelerated BCG reaction was found in 36.5 percent. Evidence of primary focus was found in 40 percent of chest radiographs. Commonest ultrasonography and computed tomography findings were mesenteric thickening, followed by intra-abdominal lymphadenopathy. Tuberculous infection could be confirmed in 38 patients. The classical plastic variety was the commonest type of ATB found. A complete cure with antituberculous drugs was documented in over 90 percent of the patients. CONCLUSION: In high prevalence zones, ATB should be considered as a differential diagnosis in children presenting with non-specific constitutional symptoms and abdominal pain. When confirmatory tests are negative or not available, supportive investigations and clinical suspicion should be considered strongly for diagnosis of ATB to avoid delay in treatment. Response to therapy in such conditions indirectly confirms diagnosis. Timely use of laparoscopy and laparotomy may be required for confirmation of diagnosis. PMID- 17909674 TI - Body composition, nutrient intake and physical activity patterns in young women during Ramadan. AB - INTRODUCTION: Muslims abstain from food and fluid between the hours of sunrise to sunset, and usually eat a large meal after sunset and a lighter meal before sunrise. The purpose of this study was to assess body composition, nutrient intake and physical activity patterns during Ramadan fasting. METHODS: This study was carried out during Ramadan in October 2004. A total of 57 female subjects were recruited from The Hashemite University in Jordan. Body weight, fat percentage, muscle mass, and percentage body water content were measured, and body mass index (BMI) was calculated. Estimated food records over a duration of three days were used to assess the intake of energy, carbohydrates, protein, fat, and sugars before and during Ramadan fasting. Physical activity patterns were determined from a three-day activity diary before and during Ramadan fasting; the amount of physical activity was expressed as the physical activity level. RESULTS: Body weight and BMI decreased significantly during Ramadan fasting. The mean energy and nutrients intake before Ramadan (energy; percent carbohydrates:protein:fat was 1,252; 56:12:33) and during Ramadan (1,171; 56:13:34) were not significantly different. The mean physical activity level was 1.54 before Ramadan and 1.51 during Ramadan, and this was also not significantly different. CONCLUSION: This study revealed that there was a significant weight loss during Ramadan. Estimates of energy, carbohydrates, protein, fat and sugar did not change, despite the reduction in the number of meals taken. The overall activity patterns remained similar. PMID- 17909675 TI - Nutrition screening among community-dwelling older adults in Singapore. AB - INTRODUCTION: This study aimed to describe responses to the DETERMINE checklist and the nutritional risk level of community-dwelling older Chinese in Singapore, aged 55 years and older. METHODS: Data was collected from a community health screening project for elderly residents in Singapore. All residents aged 55 years and older in the survey area were identified in door-to-door census surveys and were invited to participate. Participants completed a questionnaire interview conducted by research nurses. The survey also included questions which were potential predictors of nutritional risk: sociodemographic factors (age, gender, education, housing type, marital status, and living arrangement) and health related factors (self-rated health, number of medical comorbidities, hospitalisations in the past year, functional disabilities and physical health status). RESULTS: Data for analysis was provided by 2,605 Chinese subjects aged between 55 and 98 years (mean/standard deviation 66.0/7.7). The overall prevalence of nutritional risk (according to a DETERMINE score of 3 or greater) was 30.1 percent. 1,822 (69.9 percent) subjects had no nutritional risk (scores of 2 or lower), 664 (25.5 percent) had moderate nutritional risk and 119 (4.6 percent) had high nutritional risk. The most common contributions to nutritional risks were: changing food intake due to illness (40.3 percent), taking three or more different medications daily (25.0 percent), eating alone (14.5 percent) and consuming insufficient amount of fruits, vegetables or milk products on a daily basis (9.0 percent). Respondents at nutritional risk were more likely to have three or more comorbid medical conditions, were hospitalised in the past year, were functionally dependent on one or more instrumental or basic activities of daily living, were reported to have poor or fair self-rated health, and were in the lowest tertile scores for SF-12 quality of life and depression. CONCLUSION: Self-rated general health, lowered quality of life, functional disability and depression have meaningful non-circular associations with the checklist. These support the validity of the DETERMINE checklist in predicting the risk of adverse health conditions and events. PMID- 17909676 TI - Clinical and laboratory features of Nigerian patients with osteomyelitis. AB - INTRODUCTION: The aim of this study was to investigate the clinicopathological characteristics of Nigerian patients with osteomyelitis. METHODS: 30 patients with osteomyelitis and 30 apparently-healthy age- and sex-matched controls were investigated. The packed cell volume (PCV), white blood cells (WBC) and differentials, and platelet counts were measured using an automated counter, while the erythrocyte sedimentation rate (ESR) was determined by Westergren's technique. C3 activator, C1 esterase inhibitor (C1-INH), IgA, IgG and IgM were estimated by the single radial immunodiffusion method. Wound swabs, blood cultures and biopsies were taken and sent for microscopic, culture and sensitivity analysis. RESULTS: Patients with osteomyelitis had elevated total leucocytes, neutrophils, and platelet counts compared to the controls. There was also significant anaemia (t equals 3.17, p-value equals 0.002) and a significantly elevated ESR (t equals 3.75, p-value equals 0.000). Serum levels of C3 activator were significantly higher in patients with osteomyelitis (t equals 6.29, p-value equals 0.000). Although serum levels of C1-INH, IgG and IgM were higher in osteomyelitis, they were not significantly so. Serum levels of IgA were reduced in patients with osteomyelitis. Significant correlations between PCV and ESR (r equals -0.486, p-value equals 0.006), ESR and total WBC count (r equals +0.542, p-value equals 0.002), ESR and platelet count (r equals 0.445, p-value equals 0.013) and total WBC count and IgG (r equals 0.507, p-value equals 0.019) were noted . CONCLUSION: Nigerian patients with osteomyelitis have similar clinical and laboratory features already described in literature, with some noted immune dysfunctions. PMID- 17909678 TI - A review of Orang Asli newborns admitted to a neonatal unit in a Malaysian general hospital. AB - INTRODUCTION: The Orang Asli are the indigenous population in peninsular Malaysia and are in fact a diverse sub-ethnic group with different languages. Our aim was to collect data on Orang Asli newborns, from western and central Pahang, that were admitted to a general hospital with paediatric specialist services. METHODS: This is a retrospective study of all Orang Asli neonates admitted to the Neonatal Unit in Temerloh Hospital over a one-year period (2003). RESULTS: There were 65 Orang Asli admissions out of a total of 1,543 admissions to our Neonatal Unit. The average birth weight was 2,569 g. The commonest indication for admission was neonatal jaundice secondary to glucose-6-phosphate dehydrogenase deficiency. Ten babies were ventilated, seven for prematurity and three for mild-moderate perinatal asphyxia. There were three deaths: a baby with a lethal congenital abnormality, one with congenital rubella syndrome with cardiac failure, and a preterm baby delivered at 28 weeks gestation, with late neonatal sepsis. CONCLUSION: This is the first attempt to assess the health status of Orang Asli neonates in peninsular Malaysia. There are no published reports on the health status of this group of neonates. A larger multicentre study is needed to determine the exact health status of Malaysian Orang Asli newborns. PMID- 17909677 TI - Red cell autoantibodies among thalassaemia patients in Hospital Universiti Sains Malaysia. AB - INTRODUCTION: Thalassaemia is one of the major public health problems in Malaysia. Regular monthly blood transfusion remains the main treatment for severe thalassaemia patients. One of the complications of blood transfusion is the formation by the recipients of alloantibodies and autoantibodies against red blood cell (RBC) antigen. The purpose of this study was to determine the prevalence of RBC autoantibodies among multiple-transfused thalassaemic patients in our institution and factors that contribute to its development. METHODS: A prospective study was conducted in Haematology Laboratory, Hospital Universiti Sains Malaysia between January 2004 and December 2004. A total of 63 thalassaemia patients, who received regular blood transfusion were included in this study. Clinical and serological data were collected and analysed prospectively. Blood samples were subjected to standard blood bank procedures for screening of antibodies and their subsequent identification using reagent of Diamed-ID Gel microtyping system. RESULTS: There were 49 (77.8 percent) patients with Hb E/beta thalassaemia, ten (15.9 percent) beta-thalassaemia major, three (4.7 percent) Hb H Constant Spring and one (1.6 percent) Hb H disease. Only one (1.6 percent) patient had autoantibodies. There were no statistical associations found between the formation of autoantibodies with age at the start of transfusion, number of packed cell transfused and splenectomy. CONCLUSION: Our data showed a low autoimmunisation rate in multiple-transfused thalassaemia patients in our hospital. PMID- 17909679 TI - Third head of biceps brachii in an Indian population. AB - INTRODUCTION: The biceps brachii is one of the muscles of the anterior compartment of the upper arm. It is characteristically described as a two-headed muscle that originates proximally by a long head and a short head. The present study was carried out to find the occurrence of a third head of biceps brachii among a sample Indian population from the southern coastal part. METHODS: The arms of 42 cadavers were dissected and observed for variations in the origin and insertion of biceps brachii muscle bilaterally. The cadavers were embalmed and preserved in ten percent formalin. RESULTS: Among 42 arms studied, three had biceps brachii with three heads on the right side. The third head was of humeral origin, which inserted into the radial tuberosity by a common tendon with the long and short heads. The results of the present study compared with that of previous studies from medical literature shows that the occurrence of a third head of the biceps brachii muscle is relatively rare in Indians. CONCLUSION: Knowledge of the existence of the third head of biceps brachii may become significant in preoperative diagnosis and during surgery of the upper limb. PMID- 17909680 TI - Modulating multidrug resistance gene in leukaemia cells by short interfering RNA. AB - INTRODUCTION: The multidrug resistance gene, MDR1, is one of the genes responsible for resistance to chemotherapy in the treatment of leukaemia and other cancers. The discovery of RNA interference in mammalian cells has provided a powerful tool to inhibit the expression of this gene. However, very little is known about the transfection of leukaemia cells with short interfering RNA (siRNA) targeted at MDR1. This study aims to evaluate the effectiveness of two chemically-synthesised siRNA in modulating MDR1 gene and inhibiting P glycoprotein expression in leukaemic cells. We also evaluated two siRNA delivery methods in this study. METHODS: K562/Adr was transfected with two MDR1-targeted siRNA or negative control siRNA, by using cationic lipid-based transfection reagents or electroporator. Gene expression of MDR1 was quantified by real-time polymerase chain reaction and calculated as a percentage relative to the negative control siRNA. P-glycoprotein expression was evaluated via flow cytometry and drug sensitivity after treatment was assessed by cytotoxicity assays. RESULTS: The percentage of MDR1 gene knockdown from cells transfected with an electroporator was significantly higher (84.4 percent, p-value is 0.094) compared to cells transfected with cationic lipid-based transfection reagents (52.8 percent). Both siRNA significantly reduced the expression of MDR1 by 84.9 percent (p-value is 0.001) and 86.0 percent (p-value is 0.011), respectively. P glycoprotein expression was down-regulated and drug sensitivity was increased after treatment with the siRNA. CONCLUSION: This study shows that the two siRNA sequences are capable of modulating MDR1 and P-glycoprotein expressions and increased drug sensitivity. Transfection with an electroporator was superior to chemical transfection for leukaemia cells. PMID- 17909681 TI - Septic postpartum uterine inversion. AB - Puerperal uterine inversion is an uncommon but life threatening obstetrical emergency. A 26-year-old woman, para six, was referred from a peripheral hospital seven days after delivery, with a mass protruding per vaginum. Complete uterine inversion had occurred after delivery of baby and placenta. She was resuscitated and her genital infection was treated. She had a vaginal hysterectomy upon request. Her postoperative recovery was uneventful. Poor management of the third stage of labour is a common cause of uterine inversion. Early replacement of the inverted uterus is important to prevent further complications. PMID- 17909682 TI - Late postpartum eclampsia at five weeks post-delivery. AB - Postpartum eclampsia is a serious and unexpected complication. A relative increase in the incidence of postpartum eclampsia has been noted in the last few years. Late postpartum eclampsia, though initially controversial, is now recognised up to four weeks after delivery. We present such a case occurring in a 26-year-old woman. This patient, who had undergone lower segment caesarean section due to pregnancy-induced hypertension, presented with typical features of postpartum eclampsia 34 days after an asymptomatic interval. PMID- 17909683 TI - A rare variation of the profunda femoris vein in the popliteal fossa. AB - The profunda femoris artery is normally accompanied by a profunda femoris vein (deep femoral vein), which begins at the adductor magnus with various tributaries and drains into the femoral vein at the femoral triangle. Very rarely, the profunda femoris vein establishes communication with the popliteal vein. We present an anomalous profunda femoris vein in a 62-year-old male cadaver whose vein was located in the popliteal fossa as a direct communicating channel between the popliteal vein and the femoral vein. PMID- 17909684 TI - Scenario of a dirty bomb in an urban environment and acute management of radiation poisoning and injuries. AB - In the new security environment, there is a clear and present danger of terrorists using non-conventional weapons to inflict maximum psychological and economic damage on their targets. This article examines two scenarios of radiation contamination and injury, one accidental in nature leading to environmental contamination, and another of deliberate intent resulting in injury and death. This article also discusses the management of injury from radiological dispersion devices or dirty bombs, with emphasis on the immediate aftermath as well as strategy recommendations. PMID- 17909685 TI - Benign breast lesions mimicking carcinoma at mammography. AB - Many benign breast lesions pose diagnostic challenges. These lesions include abscess, haematoma, radial scar, post surgical scar, diabetic mastopathy, focal fibrosis, sclerosing adenosis, granular cell tumour, extra-abdominal desmoid tumour, medial insertion of pectoralis muscle and sternalis muscle, and axillary lymphadenopathy (due to HIV infection, collagen vascular lesions, tuberculous and bacterial lymphadenitis). Radiologists should be familiar with the characteristic imaging features of these benign lesions, and should include these benign lesions in the differential diagnosis whenever malignant-appearing findings are encountered. Correlation of the patient's clinical features with the mammographical findings and additional use of ultrasonography, fine-needle aspiration biopsy or core biopsy are helpful in establishing the final diagnosis and obviating unnecessary surgical intervention. In some of these lesions, surgery may be avoided while in others, the appropriate surgical procedure may be planned. This pictorial essay aims to illustrate the mammographical features of these lesions in a group of proven cases. PMID- 17909686 TI - Ophthalmomyiasis: a rare cause of short duration pre-septal cellulitis in a healthy non-compromised adult. PMID- 17909687 TI - The bi-directional link between women's and men's mental health. PMID- 17909690 TI - Identification of the optimal time to treat urgency after a midurethral sling procedure for stress urinary incontinence. AB - We followed 91 patients who had undergone transobturator tension-free vaginal tape procedure on their perioperative urgency symptom for 1 year to identify risk factors and optimal time to commence further treatment in the presence of postoperative urgency. Of the 59 patients with preexisting urgency, 54.2, 35.6, and 39.0% demonstrated symptom persistence at postoperative 1, 6, and 12 months, respectively. Of the 32 patients without preoperative urgency, 3.1 and 18.8% of patients demonstrated de novo urgency at 1 and 6 months, respectively, but symptom persistence to 12 months was observed in 6.2%. Overall, urgency lasting to 12 months was observed in 25 (27.5%) of the entire cohort. Preoperative urgency [p = 0.001, odds ratio (OR) 9.583] and urgency at 1 month (p = 0.001, OR 5.124) were associated with symptom persistence to 1 year after surgery. We recommend treatment if urgency is noted at 1 month postoperatively in patients with preexisting urgency, and after 6 months for those without preoperative urgency. PMID- 17909691 TI - Fibroid-induced acute urinary retention: treatment by uterine artery embolization. AB - A 39-year-old gravida 2 para 2 woman presented to our Hospital's Emergency Department with complaints of difficulty voiding. She had an enlarged leiomyomatous uterus, for which she was not receiving any current treatment. A Foley catheter placed yielded 1,500 cc of clear yellow urine; however, the patient remained Foley-dependent for 2 weeks until she underwent uterine artery embolization (UAE). Twenty-four hours afterwards, the Foley catheter was removed and the patient spontaneously voided with negligible post-void residual. There was no recurrence of urinary retention or development of any other urinary symptoms during the outpatient follow-up period. Repeat pelvic magnetic resonance imaging (MRI) 1 week after UAE showed negligible reduction in the size of the fibroids and uterus in comparison with a pre-procedure MRI. The Vascular Steal Theory, first presented in this paper, discusses this improvement in symptoms without significant change in size. PMID- 17909693 TI - Adaptive evolution in an avian reproductive protein: ZP3. AB - Proteins involved in reproduction appear to be evolving adaptively across taxa. This rapid evolution is thought to be the result of forces involved in sexual selection. One of the most often suggested of these forces is sexual conflict involving sperm competition and polyspermy avoidance. Bird species offer a unique opportunity to test this hypothesis since the avian egg coat tolerates physiological polyspermy, or the penetration of multiple sperm during fertilization, without negative effects on later development. Despite this, and the extensive amount of data gathered on sexual selection in birds, there are limited studies on the patterns of evolution of avian reproductive proteins. Here we present an analysis of the pattern of evolution of Zona Pellucida 3 (ZP3), a protein present on the avian egg coat. We found that, across several galliform and a single anseriform species, ZP3 appears to be diverging by positive adaptive evolution. In an exploratory analysis of portions of the gene in Callipepla californica we also found evidence of a selective sweep at the putative sperm binding region of the protein. In sum, ZP3 in birds, like reproductive proteins in other species, appears to be adaptively evolving. This result suggests that polyspermy avoidance is not sufficient to explain positive Darwinian selection in reproductive proteins across taxonomic groups. Clearly, the inclusion of bird species in the study of reproductive proteins across taxa promises to add greatly to the discussion of the factors driving the widespread phenomenon of adaptive evolution in reproductive proteins. PMID- 17909694 TI - Genomic evolution of the proteasome system among hemiascomycetous yeasts. AB - Components of the proteasome-ubiquitin pathway are highly conserved throughout eukaryotic organisms. In S. cerevisiae, the expression of proteasomal genes is subject to concerted control by a transcriptional regulator, Rpn4p, interacting with a highly conserved cis-regulatory element, PACE, located in the upstream regions of these genes. Taking advantage of sequence data accumulated from 15 Hemiascomycetes, we performed an in silico study to address the problem of how this system might have evolved among these species. We found that in all these species the Rpn4p homologues are well conserved in terms of sequence and characteristic domain features. The "PACE patterns" turned out to be nearly identical among the Saccharomyces "sensu stricto" species, whereas in the evolutionary more distant species the putatively functional cis-regulatory motifs revealed deviations from the "canonical" PACE nonamere sequence in one or two nucleotides. Our findings suggest that during evolution of the Hemiascomycetes such slightly divergent ancestral motifs have converged into a unique PACE element for the majority of the proteasomal genes within the most recent species of this class. Likewise, the Rpn4 factors within the most recent species of this class show a higher degree of similarity in sequence than their ancestral counterparts. By contrast, we did not detect PACE-like motifs among the proteasomal genes in other eukaryotes, such as S. pombe, several filamentous fungi, A. thaliana, or humans, leaving the interesting question which type of concerted regulation of the proteasome system has developed in species other than the Hemiascomycetes. PMID- 17909692 TI - An ancient repeat sequence in the ATP synthase beta-subunit gene of forcipulate sea stars. AB - A novel repeat sequence with a conserved secondary structure is described from two nonadjacent introns of the ATP synthase beta-subunit gene in sea stars of the order Forcipulatida (Echinodermata: Asteroidea). The repeat is present in both introns of all forcipulate sea stars examined, which suggests that it is an ancient feature of this gene (with an approximate age of 200 Mya). Both stem and loop regions show high levels of sequence constraint when compared to flanking nonrepetitive intronic regions. The repeat was also detected in (1) the family Pterasteridae, order Velatida and (2) the family Korethrasteridae, order Velatida. The repeat was not detected in (1) the family Echinasteridae, order Spinulosida, (2) the family Astropectinidae, order Paxillosida, (3) the family Solasteridae, order Velatida, or (4) the family Goniasteridae, order Valvatida. The repeat lacks similarity to published sequences in unrestricted GenBank searches, and there are no significant open reading frames in the repeat or in the flanking intron sequences. Comparison via parametric bootstrapping to a published phylogeny based on 4.2 kb of nuclear and mitochondrial sequence for a subset of these species allowed the null hypothesis of a congruent phylogeny to be rejected for each repeat, when compared separately to the published phylogeny. In contrast, the flanking nonrepetitive sequences in each intron yielded separate phylogenies that were each congruent with the published phylogeny. In four species, the repeat in one or both introns has apparently experienced gene conversion. The two introns also show a correlated pattern of nucleotide substitutions, even after excluding the putative cases of gene conversion. PMID- 17909695 TI - Avian UCP: the killjoy in the evolution of the mitochondrial uncoupling proteins. AB - The understanding of mitochondrial functioning is of prime importance since it combines the production of energy as adenosine triphosphate (ATP) with an efficient chain of redox reactions, but also with the unavoidable production of reactive oxygen species (ROS) involved in aging. Mitochondrial respiration may be uncoupled from ATP synthesis by a proton leak induced by the thermogenic uncoupling protein 1 (UCP1). Mild uncoupling activity, as proposed for UCP2, UCP3, and avian UCP could theoretically control ROS production, but the nature of their transport activities is far from being definitively understood. The recent discovery of a UCP1 gene in fish has balanced the evolutionary view of uncoupling protein history. The thermogenic proton transport of mammalian UCP1 seems now to be a late evolutionary characteristic and the hypothesis that ancestral UCPs may carry other substrates is tempting. Using in silico genome analyses among taxa and a biochemical approach, we present a detailed phylogenetic analysis of UCPs and investigate whether avian UCP is a good candidate for pleiotropic mitochondrial activities, knowing that only one UCP has been characterized in the avian genome, unlike all other vertebrates. We show, here, that the avian class seems to be the only vertebrate lineage lacking two of the UCP1/2/3 homologues present in fish and mammals. We suggest, based on phylogenetic evidence and synteny of the UCP genes, that birds have lost UCP1 and UCP2. The phylogeny also supports the history of two rounds of duplication during vertebrate evolution. The avian uncoupling protein then represents a unique opportunity to explore how UCPs' activities are controlled, but also to understand why birds exhibit such a particular relationship between high metabolism and slow rate of aging. PMID- 17909696 TI - Tumor necrosis factor-alpha antagonism protects from myocardial inflammation and fibrosis in experimental diabetic cardiomyopathy. AB - To investigate the effect of anti-cytokine-based therapy in the course of diabetic cardiomyopathy, we performed a study using an anti-TNF-alpha monoclonal antibody treatment (mab) in Sprague male Dawley (SD) rats with streptozotocin induced diabetic cardiomyopathy. Five days after streptozotocin injection, rats were treated with the anti-TNF-alpha mAb C432A for 6 weeks.At the end of the study, left ventricular (LV) function was determined by a pressure-catheter. Intercellular adhesion molecule (ICAM)-1, vascular adhesion molecule (VCAM)-1, beta2-lymphocyte-integrins(+) (CD18(+), CD11a(+), CD11b(+)), ED1/CD68(+) and cytokine (TNF-alpha, interleukin (IL)-1beta)- expressing infiltrates, total collagen content and stainings of collagen I and III were quantified by digital image analysis. LV phosphorylated and total ERK protein levels were determined by Western Blot. TNFalpha-antagonism reduced ICAM-1- and VCAM-1 expression and leukocyte infiltration to levels of non-diabetics and decreased macrophage residence by 3.3-fold compared with untreated diabetics. In addition, anti-TNF alpha mAb-treatment decreased diabetes-induced cardiac TNF-alpha and IL-1beta expression by 2.0-fold and 1.8- fold, respectively, and reduced the ratio of phosphorylated to total ERK by 2.7-fold. The reduction in intramyocardial inflammation was associated with a 5.4-fold and 3.6-fold reduction in cardiac collagen I and III content, respectively. This was reflected by a normalization of cardiac total collagen content to levels of non-diabetics and associated with an improved LV function. TNFalpha-antagonism attenuates the development of experimental diabetic cardiomyopathy associated with a reduction of intramyocardial inflammation and cardiac fibrosis. PMID- 17909697 TI - A case of pancytopenia and splenomegaly: haematological disease? PMID- 17909698 TI - Shirt fold mimicking pneumothorax on chest radiograph: accurate diagnosis by ultrasound. PMID- 17909699 TI - Late-onset rhabdomyolysis in pneumococcal meningitis: a case report. PMID- 17909700 TI - Patients with an intermediate or high risk of a pulmonary embolism continue to pose a diagnostic challenge. PMID- 17909702 TI - Rhabdomyolysis. AB - Rhabdomyolysis is a syndrome involving the breakdown of skeletal muscle causing myoglobin and other intracellular proteins and electrolytes to leak into the circulation. The development of rhabdomyolysis is associated with a wide variety of diseases, injuries, medications and toxins. While the exact mechanisms responsible for all the causes are not fully understood, it is clear that muscle damage can occur from direct injury or by metabolic inequalities between energy consumption and energy production. Rhabdomyolysis is diagnosed by elevations in serum creatine phosphokinase (CPK), and while there is no established serum level cut-off, many clinicians use five times the upper limit of normal ( approximately 1000 U/l). Rhabdomyolysis can be complicated by acute renal failure (occurring in 4%-33% of patients), compartment syndrome, cardiac dysrhythmias via electrolyte abnormalities, and disseminated intravascular coagulopathy. The mainstay of treatment is hospitalisation with aggressive intravenous fluid (IVF) resuscitation with the correction/prevention of electrolyte abnormalities. There are additional adjunctive therapies to IVF, such as alkalinisation of the urine with sodium bicarbonate, diuretic therapy or combinations of both; however the lack of large randomised control studies concerning the benefits of these treatments makes it difficult to make strong recommendations for or against their use in the treatment of rhabdomyolysis. Regardless of these controversies, the overall prognosis for rhabdomyolysis is favourable when treated with early and aggressive IVF resuscitation, and full recovery of renal function is common. Irrespective of the cause of rhabdomyolysis the mortality rate may still be as high as 8%. This is a comprehensive review of the pathophysiology, diagnosis, complications and treatment options for rhabdomyolysis. PMID- 17909703 TI - Aortic coarctation in the elderly: how many errors lie behind an unexpected diagnosis? PMID- 17909704 TI - Seizures, headache and thrombocytopenia: diagnosis and treatment do not always come in a standard sequence. PMID- 17909705 TI - Triple therapy of warfarin, aspirin and a thienopyridine for patients treated with vitamin K antagonists undergoing coronary stenting. A review of the evidence. AB - Dual antiplatelet treatment of aspirin and a thienopyridine (either ticlopidine or clopidogrel) is the standard of care in patients undergoing coronary artery stenting (PCI-S). Such treatment however, is not generally applicable in patients with concomitant indication for vitamin K antagonists (VKA), in whom therefore the optimal treatment is currently undefined. According to the limited available evidence, the management of these patients is substantially variable, but triple therapy of VKA, aspirin and a thienopyridine is the most frequently adopted. Both VKA and dual antiplatelet treatment in fact are warranted to actually prevent systemic thromboembolism and stent thrombosis, although an increased haemorrhagic risk might be associated with such therapy. A substantial incidence of bleeding has been effectively observed with triple therapy in a few, small, retrospective, observational series. The risk of haemorrhage appears to increase with the duration of treatment, although concomitant factors (i.e., advanced age, presence of gastrointestinal lesions, excessive anticoagulation or traumatic manoeuvres), rather than the administration of numerous antithrombotic agents in itself, may play a role. As expected, no thromboembolic or thrombotic events have been generally reported with such treatment. Because of the limited and poor quality data currently available on the management of patients with an indication for VKA undergoing PCI-S, large-scale registries and clinical trials are warranted to determine the optimal antithrombotic treatment in this patient subset, which is foreseen to progressively increase over the next years. PMID- 17909706 TI - Triple antithrombotic therapy with aspirin, clopidogrel and warfarin--a persisting dilemma. PMID- 17909708 TI - Norrisiella sphaerica gen. et sp. nov., a new coccoid chlorarachniophyte from Baja California, Mexico. AB - A new chlorarachniophyte, Norrisiella sphaerica S. Ota et K. Ishida gen. et sp. nov., from the coast of Baja California, Mexico is described. We examined its morphology, ultrastructure, and life cycle in detail, using light microscopy, transmission electron microscopy, and time-lapse videomicroscopy. We found that this chlorarachniophyte possessed the following characteristics: (1) vegetative cells were coccoid and possessed a cell wall, (2) a pyrenoid was slightly invaded by plate-like periplastidial compartment from the tip of the pyrenoid, (3) a nucleomorph was located near the pyrenoid base in the periplastidial compartment, (4) cells reproduced vegetatively via autospores, and (5) a flagellate stage was present in the life cycle. This combination of characteristics differs from any of the described chlorarachniophyte genera, and therefore a new genus is established. Fluorescent microscopic observations suggested that the alga formed multinucleate cells prior to forming autospores. Time-lapse observations during autospore formation showed that cytokinesis occurred simultaneously in the multinucleate cells. Zoospores were also produced, and video sequences captured the release of zoospores from coccoid cells. PMID- 17909709 TI - Treatment strategy for hilar cholangiocarcinoma, with special reference to the limits of ductal resection in right-sided hepatectomies. AB - The surgical anatomy of the hepatic hilar region is characterized by the three dimensional formation of the branches of the bile duct, portal vein, and hepatic artery. The limit of ductal resection in hepatectomy for hilar cholangiocarcinoma is the most peripheral point where the hepatic ducts can be separated from the vasculature. The limit is different for each type of hepatectomy because the portal vein branches that should be preserved or divided vary with the extent of the hepatectomy, and therefore the limit of separation of the hepatic ducts differs. Surgeons are required to understand the surgical anatomy and to identify the precise area of cancer spread on a preoperative cholangiogram so as to choose the appropriate type of hepatectomy, and to ensure that the remnant ductal margin is cancer-negative. PMID- 17909710 TI - Preoperative assessment of hilar cholangiocarcinoma by multidetector row computed tomography. AB - BACKGROUND/PURPOSE: Hilar cholangiocarcinoma is the one of the most difficult carcinomas to diagnose because of the localization of the main tumor at the hepatic hilus, and because of the complex anatomy of the biliary, artery, and portal systems. To perform a curative operation, it is important to evaluate the extent of carcinoma and the resectability. Hilar cholangiocarcinoma often extends along the axis of the bile duct. Percutaneous transhepatic cholangiogaraphy (PTC) and/or endoscopic retrograde cholangiography (ERC) are usually performed to diagnose the extent of the hilar cholangiocarcinoma. However, computed tomography (CT) was thought not to be useful because its resolution is poor. Now that multidetector row CT (MDCT) and high-performance imaging systems are available, the diagnostic strategy for hilar cholangiocarcinoma has changed. METHODS: In this study, we analyzed the preoperative diagnostic imaging of 24 consecutive patients whose hilar cholangiocarcinoma was confirmed by histopathological examination. All patients were submitted to 16-channel MDCT, except for those with an allergy to iodine contrast medium. The data obtained from MDCT were analyzed and checked by both radiologists and surgeons, using multiplanar reconstruction (MPR) images. RESULTS: The accuracy of diagnosis of horizontal spreading was 80.9% and that of vertical spreading was 100%. However, the sensitivity for lymph node metastasis was insufficient. Based on the data from MDCT and other examinations, all patients underwent surgery. Curative operation was performed in 15 patients (62.5%). CONCLUSIONS: Our results indicate that 16 channel MDCT is reliable for the diagnosis of hilar cholangiocarcinoma, especially prior to bile duct drainage. Thus, it is important to perform MDCT when patients with obstructive jaundice are encountered. PMID- 17909711 TI - Preoperative biliary drainage for hilar cholangiocarcinoma. AB - Hilar cholangiocarcinomas grow slowly, and metastases occur late in the natural history. Surgical cure and long-term survival have been demonstrated, when resection margins are clear. Preoperative biliary drainage has been proposed as a way to improve liver function before surgery, and to reduce post-surgical complications. Percutaneous transhepatic biliary drainage (PTBD) with multiple drains was previously the preferred method for the preoperative relief of obstructive jaundice. However, the introduction of percutaneous transhepatic portal vein embolization (PTPE) and wider resection has changed preoperative drainage strategies. Drainage is currently performed only for liver lobes that will remain after resection, and for areas of segmental cholangitis. Endoscopic biliary drainage (EBD) is less invasive than PTBD. Among EBD techniques, endoscopic nasobiliary drainage (ENBD) is preferable to endoscopic biliary stenting (EBS), because secondary cholangitis (due to the retrograde flow of duodenal fluid into the biliary tree) does not occur. ENBD needs to be converted to PTBD in patients with segmental cholangitis, those with a prolonged need for drainage, or when the extent of longitudinal tumor extension is not sufficiently well characterized. PMID- 17909707 TI - Adiponectin and the cardiovascular system: from risk to disease. AB - Adiponectin is known to play a role in fatty acid and glucose metabolism through a change in insulin sensitivity and activation of fuel oxidation by AMP-activated protein kinase. Adiponectin can be considered an important factor able to modulate the adipovascular axis which, through genomic and environmental influences, affects the cardiovascular risk milieu, from the pre-metabolic syndrome-- through the metabolic syndrome--to the overt atherosclerotic process and its clinical manifestations. Hypoadiponectinaemia can be viewed as an early sign of a complex cardiovascular risk factor predisposing to the atherosclerosis process as well as a contributing factor accelerating the progress of the atherosclerotic plaque. In addition, adiponectin per se holds a protective role thanks to its anti-inflammatory and antiatherogenic properties. The early identification of patients "at cardiovascular risk" means in the current practice to search for indexes of metabolic derangements and pro-inflammatory status (adiponectin) from adolescence and childhood. PMID- 17909712 TI - Recent advances in the treatment of hilar cholangiocarcinoma: portal vein embolization. AB - The clinical application of portal vein embolization (PVE) has contributed to improving the postoperative outcome of hilar cholangiocarcinoma. The enlarged nonembolized lobe after PVE protects the patient from postoperative hepatic failure, due to the increased functional reserve, and shortens the hospital stay. Although numerous reports have shown beneficial effects of PVE on postoperative outcome after extended hepatectomy, no randomized controlled study has been performed so far. It is urgent to establish a "gold standard" of PVE, because the indications, approach to the portal vein, types of embolic materials, and methods used to evaluate the function of the future liver remnant are variable among institutions. The indications and procedures of PVE for hilar cholangiocarcinoma may be different from those for hepatocellular carcinoma or colorectal metastasis, because, in many patients with hilar cholangiocarcinoma, biliary cancer is associated with biliary obstruction and cholangitis. This review article summarizes the contribution of PVE to the outcome of postoperative management in patients with hilar cholangiocarcinoma needing extended hepatectomy. We also describe our PVE procedure, which has been established from our experience of more than 240 cases of biliary cancer. Furthermore, the drawbacks of PVE, which may reduce the pool of candidates for surgery, are also discussed. PMID- 17909713 TI - Changing trends in surgical outcomes after major hepatobiliary resection for hilar cholangiocarcinoma: a single-center experience over 25 years. AB - BACKGROUND/PURPOSE: Hepatobiliary resection (HBR) for hilar cholangiocarcinoma (HCCa) remains a technically demanding procedure and is still associated with significant rates of morbidity and mortality. The aim of this study was to characterize changes in surgical outcomes following major HBR for HCCa at a single center over a 25-year period. METHODS: Between 1980 and 2004, 126 patients undergoing preoperative biliary drainage, portal vein embolization, and major HBR were enrolled in this study. The patients were divided into two groups according to the chronological treatment period; i.e., patients who underwent surgery during the initial 20-year period (1980-1999; early group [EG]) and those who underwent surgery during the most recent 5-year period (2000-2004; late group [LG]). Clinicopathological variables were compared retrospectively between the two groups. RESULTS: The mortality rate improved from 7.9% in the EG to 0% in the LG, but this difference did not reach the level of statistical significance (P = 0.058). The overall survival rate at 1, 3, and 5 years was 82.4%, 43.9%, and 35.2%, respectively. The overall survival rate was similar in the two groups (P = 0.153). Morbidity was documented in 57.1% of all the patients, and was comparable in the two groups (P = 0.471), but the rate of major morbidity was significantly higher in the EG (P = 0.031). Red blood cell and fresh frozen plasma transfusion requirements were significantly reduced in the LG, both in regard to the number of patients and the amount of blood product administered. The mean length of postoperative hospital stay was significantly reduced, from 74.4 + -56.3 days in the EG to 29.0 + -11.8 days in the LG (P < 0.001). Sixty-nine patients (54.8%) had stage III or IV disease (according to the General rules for surgical and pathological studies on cancer of the biliary tract of the Japanese Society of Biliary Surgery), and 55 patients (43.7%) showed positive surgical margins. There were no differences between the two groups in terms of surgical margins or pathological staging. CONCLUSIONS: Improvements were documented in rates of major morbidity, length of hospital stay, and the mortality rate in the LG when compared with the EG. The overall survival rate was similar in the two groups. Blood transfusion requirements were significantly reduced in the LG when compared with the EG. However, the high proportion of patients with positive surgical margins remains a significant problem. PMID- 17909714 TI - Recent advance in the treatment of hilar cholangiocarcinoma: hepatectomy with vascular resection. AB - Radical surgical resection has been revealed to be the only hope of cure for the patient with hilar cholangiocarcinoma. Therefore, major efforts have been made to increase the resection rate by surgeons employing combined hepatic resection and vascular resection of the portal vein and the hepatic artery. Especially, the technical feasibility and surgical safety of hepatic resection with combined portal vein resection have recently been reported by several authors. On the other hand, there have been few reports of combined hepatic artery resection in hilar cholangiocarcinoma. There are fears that combined vascular resection with extended hepatectomy for hilar cholangiocarcinoma may lead to high surgical morbidity and mortality. Herein, we describe the results of aggressive surgical approaches in our series, and we also review the outcomes of hepatic resection with combined vascular resection in the previously reported literature. PMID- 17909715 TI - Liver transplantation for hilar cholangiocarcinoma. AB - Hilar cholangiocarcinoma was accepted as an indication for liver transplantation at the beginning of the transplantation era. Owing to disappointing long-term results for this indication, and in parallel, encouraging results in patients with benign disease, hilar cholangiocarcinoma has generally not been accepted as an indication for liver transplantation in recent years. To improve results, more aggressive approaches have been used: "abdominal organ cluster transplantation" and "extended bile duct resection", which lead to increased long-term survival rates. However, with improving results after conventional extrahepatic bile duct resection in combination with partial hepatectomy, extended procedures in combination with liver transplantation never became a real option in the treatment of hilar cholangiocarcinoma. However, new awareness of liver transplantation in the treatment of this cancer has been raised for patients with hilar cholangiocarcinoma in the context of underlying liver diseases such as primary sclerosing cholangitis, which preclude liver resection. Current results show increased survival figures, in particular in well-selected patients with early tumor stages. Further improvements in long-term survival may be reached with new adjuvant and neoadjuvant protocols. Patients with neoadjuvant radiochemotherapy show long-term results similar to those for liver transplantation for other indications. Also, photodynamic therapy and the use of new antiproliferative immunosuppressive agents may be an approach for further improvement of the long-term results. Currently, liver transplantation for the treatment of hilar cholangiocarcinoma should be restricted to centers with experience in the treatment of this cancer and should be taken into consideration in patients with contraindications to liver resection. PMID- 17909716 TI - Bile duct injury repair: when? what? who? AB - Laparoscopic cholecystectomy is associated with a two-to-four times higher risk of bile duct injury (BDI) than open cholecystectomy. BDI can lead to significant morbidity and even mortality. The first priority in BDI is to control peritoneal and biliary sepsis and to convert an acute BDI to a controlled external biliary fistula (EBF) - this can be achieved by endoscopic and/ or radiological intervention in most cases. This should be followed by assessment of the extent of injury - both biliary and vascular. Immediate management of BDI recognized during cholecystectomy depends on the type of injury, the condition of the patient, and the experience of the surgeon. For BDI recognized after cholecystectomy, early repair is not recommended, as the results are poor. The EBF may evolve into a benign biliary stricture (BBS), which should be electively repaired by a Roux-en-Y hepatico-jejunostomy. The use of an endoscopic stent as definitive management of BDI is not recommended. Long-term follow-up is essential after the repair of a BBS, as recurrence can occur several years after repair. Recurrent BBS is best treated with endoscopic balloon dilatation. Excellent early and long-term results can be obtained in specialized units at tertiary care referral centers. PMID- 17909717 TI - Dissection of the uncinate process and pancreatic head behind the portal vein using endovascular staplers. AB - Pancreatic resections have evolved into safe operations in experienced centers. Technical refinements continue to further improve operating time, intraoperative blood loss, and outcome after these procedures. The dissection of the uncinate process/pancreatic head is one of the critical steps during pancreaticoduodenectomy. This step can be time-consuming, with the possibility of troublesome hemorrhage. This article describes, in a stepwise fashion, the use of endovascular stapler devices that facilitate uncinate process/pancreatic head dissection during pancreaticoduodenectomy. PMID- 17909718 TI - Preoperative high-dose steroid administration attenuates the surgical stress response following liver resection: results of a prospective randomized study. AB - BACKGROUND/PURPOSE: Major abdominal surgery such as liver resection is associated with an excessive hyperinflammatory response and transient immunosuppression. We investigated the immunomodulating effect of preoperative pulse administration of high-dose methylprednisolone in patients undergoing hepatic resection without pedicle clamping. METHODS: Twenty patients who underwent hepatic resection were randomized into two groups: a steroid group (n = 10), in which patients were given 30 mg/kg per body weight (BW) methylprednisolone intravenously, and a control group (n = 10), in which patients received a placebo (sodium chloride) infusion. The main outcome parameter to assess systemic stress was the serum plasma level of interleukin-6 (IL-6). To evaluate cell-mediated immune function, human leukocyte antigen-DR (HLA-DR) expression on peripheral blood monocytes and lipopolysaccharide (LPS)-induced tumor necrosis factor-alpha (TNF-alpha) release by peripheral monocytes was measured. Other investigated serum parameters included C-reactive protein (CRP), total bilirubin, alanine aminotransferase (ALT), prothrombin time (PT)-INR, and cytokines such as IL-8 and IL-10 and TNF alpha. Postoperative convalescence, complication rate, and length of hospital stay were compared between the groups. RESULTS: Postoperative plasma concentrations of IL-6 (days 1 and 2), IL-8 (days 2 and 3), and CRP (days 1-4) were significantly lower in the steroid than in the control group. The total bilirubin concentration was significantly lower on day 6 in the steroid than in the control group. Four hours after surgery, LPS-induced TNF-alpha secretion was significantly reduced in the steroid group, but it increased rapidly during the following days. HLA-DR, ALT, and PT-INR levels were not different between the two groups. The postoperative hospital stay in the steroid group was significantly lower compared to that in the control group (mean, 10.5 days versus 14.8 days; P < 0.05). No differences were found in the convalescence score or postoperative complication rate. CONCLUSIONS: Intravenous methylprednisolone administration before hepatic resection significantly reduced systemic inflammatory cytokine release. No adverse effect on immunity was noted due to the methylprednisolone. We found no significant difference in the convalescence score, but a significantly shorter hospital stay in the steroid group. Further studies with more patients are needed to elucidate the clinical impact of preoperative steroid bolus therapy in liver surgery. PMID- 17909719 TI - Green-tea polyphenol (-)-epigallocatechin-3-gallate provides resistance to apoptosis in isolated islets. AB - BACKGROUND/PURPOSE: Apoptosis resulting from disruption of the normal cell-matrix relationship (anoikis) during islet isolation, and the reactive oxygen and nitrogen species generated following hypoxia/reoxygenation (H/R) can lead to a loss of islet tissue in culture and the reduced survival of transplanted pancreatic islets. The aim of this study was to investigate the effect of (-) epigallocatechin-3-gallate (EGCG), a well-known antiapoptotic agent, on inhibiting anoikis and H/R injury in an in vitro islet culture system. METHODS: Islets were isolated from F344 rats and cultured under normal or H/R condition with/without EGCG. RESULTS: EGCG inhibited apoptosis and lactate-dehydrogenase leakage from anoikis and H/R in a dose-dependent manner. Further, EGCG prevent increases in 8-hydroxy-2'-deoxyguanosine content and inhibited the decline of insulin secretory function induced by H/R. CONCLUSIONS: These results suggest that the addition of EGCG to an islet culture system may improve the survival rate of isolated islets and reduce the loss of functional islet mass that compromises the stable reversal of diabetes after islet transplantation. PMID- 17909720 TI - Choledocho-choledochostomy: the natural history of healing in pigs. AB - BACKGROUND/PURPOSE: Aiming to investigate the natural history of the healing of choledocho-choledochostomies. METHODS: Fifty-five female pigs of 57 kg median weight were used for the experiments. The gallbladder was removed and the common bile duct transected. Continuity was re-established by standardized single-line, interrupted, and inverted sutures. The pigs had a planned postoperative survival of up to 14 days with a subsequent laparotomy for evaluation. Blood samples were drawn prior to the first and the final operations. During laparotomy the animals were investigated for signs of cholascos, and an intraoperative cholangiography was performed. The excised anastomosis was examined for breaking strength and collagen content. RESULTS: Standard liver parameters were not significantly affected by the surgery, and cholangiography showed no signs of extrahepatic stenosis or intrahepatic dilatation. Breaking strength showed a decrease for the initial 3 postoperative days (PODs), then an increase to a stable level on PODs 6 to 14. Collagen content per volume showed a rise on PODs 0 to 1, then no change until POD 4, followed by a gradual rise until day 6. Subsequently a stable level was reached until POD 14. Two pigs were excluded due to minor cholascos. CONCLUSIONS: The present study on pigs shows that choledocho-choledochostomies, judged by breaking strength and collagen content, regain a stable level of strength 6 days after operation. PMID- 17909721 TI - Treatment outcome of selective digestive decontamination and enteral nutrition in patients with severe acute pancreatitis. AB - BACKGROUND/PURPOSE: Sepsis due to infected pancreatic necrosis is the most serious complication in the late phase of severe acute pancreatitis (SAP). Bacterial translocation from the gut is thought to be the main cause of pancreatic infection. The possibility has recently been reported that selective digestive decontamination (SDD) and enteral nutrition (EN) may alleviate the complications and reduce the mortality rate in patients with SAP. We analyzed the treatment outcome of SDD and EN in patients with SAP. METHODS: We divided 90 patients with SAP into three groups: SDD(-)EN(-),group A; SDD(+)EN(-), group B; and SDD(+)EN(+), group C. Clinical outcome was analyzed retrospectively. The effect of SDD was compared in groups A and B, and the effect of EN was compared in groups B and C. RESULTS: The background of patients was not significantly different between the groups. SDD reduced the incidence of organ dysfunction (from 70% to 59%) and the mortality rate (from 40% to 28%), but the differences were not significant. EN reduced the incidence of infected pancreatic necrosis (from 31% to 24%) and the frequency of surgery for pancreas (from 28% to 18%), and further reduced the mortality rate (from 28% for SDD to 16%), but the differences were not significant. The peripheral lymphocyte count was significantly increased in patients with EN. CONCLUSIONS: SDD and EN did not significantly affect the treatment outcome in SAP. However, the results in this study raise the possibility that SDD and EN may decrease the complications and reduce the mortality rate in SAP. The efficacy of SDD and EN for SAP should be evaluated in a randomized controlled trial. PMID- 17909722 TI - Protective effects of a hibernation-inducer on hepatocyte injury induced by hypothermic preservation. AB - BACKGROUND/PURPOSE: For hepatocyte-based cell therapy to be realistic, the method chosen for cryopreservation or hypothermic preservation is critical. The aim of the present study was to clarify whether D-Ala2-Leu5-enkephalin (DADLE), a hibernation inducer, has protective effects on hepatocytes with regard to hypothermic preservation injury. METHODS: A suspension of rat hepatocytes was stored at 4 degrees C for 24 h with or without DADLE. Their viability was measured by the trypan blue dye exclusion method, and alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) levels in the preservation solution were measured. After 24 h of cold storage, viable hepatocytes were cultured at 37 degrees C for another 24 h. Then albumin production and lidocaine clearance were measured. RESULTS: DADLE significantly improved the survival rate of hepatocytes. The levels of ALT and LDH in the preservation solution with DADLE were significantly lower than those in the preservation solution without DADLE. The treated viable hepatocytes maintained both albumin synthesis and lidocaine clearance. CONCLUSIONS: DADLE appears to have protective effects on hepatocytes with regard to hypothermic preservation injury in vitro. This hibernation-inducer is useful in prolonged hypothermic preservation for hepatocyte-based therapy. PMID- 17909723 TI - Management of a stenotic pancreatico-digestive tract anastomosis following pancreatoduodenectomy. AB - Early postoperative complications of pancreatico-digestive anastomosis following pancreatoduodenectomy are pancreatic fistula and pancreatitis affecting the pancreatic tail. Stenosis of the anastomosis is a later complication. Symptomatic and painful presentations are difficult to treat, and the optimal treatment is not currently defined. The aim of this work was to retrospectively report two cases of pancreaticogastrostomy stenosis. In both patients, the complication was diagnosed, with pancreatitis that developed following pancreatoduodenectomy. These patients were treated surgically, by fashioning a new anastomosis. Pancreaticogastrostomy has been viewed as a simpler and more secure reconstruction technique, with a lower occurrence rate of pancreatic fistula, than that of pancreaticojejunostomy. One complication of this surgery, however, is stenosis of the anastomosis. Following pancreatoduodenectomy, stenosis of the pancreaticogastrostomy may not occur until many years later. In a significant percentage of patients it is without clinical signs. It may be discovered after systematic explorations of patients following pancreaticogastrostomy or pancreaticojejunostomy. There is no study regarding the optimal treatment of postoperative stenosis of a pancreatico-gastric anastomosis. We believe that the optimal treatment is surgical. The intervention involves resection of the stenosis, and the formation of a new anastomosis. PMID- 17909724 TI - Mucinous carcinoma of Vater's ampulla with a unique extension along the main pancreatic duct. AB - We report a case of mucinous carcinoma of Vater's ampulla with a unique extension along only the main pancreatic duct (MPD) and microinvasion to the pancreas. A 52 year-old man was referred to our hospital for the evaluation and treatment of acute pancreatitis. Abdominal computed tomography (CT) demonstrated swelling in the head of the pancreas with a mass in the duodenum. Hypotonic duodenography and endoscopic examination revealed a well-defined mass, measuring about 25 mm in size, in Vater's ampulla. A biopsy specimen of the tumor showed moderately differentiated adenocarcinoma. A pylorus-preserving pancreaticoduodenectomy with a regional lymphadenectomy was performed, under a preoperative diagnosis of adenocarcinoma of Vater's ampulla with direct invasion into the head of the pancreas. The resected specimen of the duodenum confirmed the presence of the mass, which measured 22 x 15 mm in size, in Vater's ampulla. Microscopically, the tumor consisted of two components: moderately differentiated adenocarcinoma in the peripheral region of the tumor Vater's papilla and mucinous carcinoma in the central region of the tumor. The mucinous carcinoma component uniquely extended along only the MPD with microinvasion to the pancreas. Immunohistochemically, both the moderately differentiated adenocarcinoma and the mucinous carcinoma were positive for cytokeratin 20 (CK20) and negative for cytokeratin 7 (CK7) which is the pattern of intestinal-type carcinoma of Vater's ampulla. We concluded that the original site of this tumor may have been the duodenal epithelium of Vater's ampulla originally moderately differentiated adenocarcinoma-which subsequently changed to mucinous carcinoma that extended along only the MPD with microinvasion to the pancreas. PMID- 17909725 TI - Intraductal papillary-mucinous adenocarcinoma in the remnant pancreas after pancreatoduodenectomy for cancer of Vater's papilla associated with intraductal papillary-mucinous adenoma. AB - A 72-year-old woman, who had undergone pylorus-preserving pancreatoduodenectomy 3 years before for cancer of Vater's papilla associated with a branch-type intraductal papillary-mucinous adenoma (IPMA), developed dilatation of the main duct and a nodular lesion in the remnant pancreas. Total pancreatectomy was performed, which revealed that the lesion was intraductal papillary-mucinous adenocarcinoma (IPMC) with minimal invasion, suggesting the metachronous multicentric occurrence of this intraductal papillary-mucinous neoplasm (IPMN). Because there were no malignant cells at the pancreaticojejunostomy, and because the histological appearance of the main-duct IPMC was different from that of the IPMA in the primary specimen, the main-duct IPMC was thought to be of different origin from the IPMA. These findings suggest that careful surveillance of the gastrointestinal tract and careful follow up are necessary for IPMN, because an IPMN could be associated with other gastrointestinal tract malignancies. PMID- 17909726 TI - Large periampullary villous tumor of the duodenum. AB - A 67-year-old woman, who had symptoms of epigastric pain and abdominal distension, was found, on endoscopy, to have a large sessile villous adenoma of the periampullary duodenum. Despite the lack of evidence of malignancy, a pancreaticoduodenectomy procedure was performed, mainly because of the tumor size and site, involving the ampulla of Vater. The presence of the carcinoma was diagnosed only in the resected specimen by definitive histology. Because there is no general consensus on the optimal surgical procedure for the treatment of villous tumors of the duodenum, especially for the early stages, the indications for the operative procedure are discussed, based on a review of the literature. PMID- 17909727 TI - Gallbladder carcinoma associated with occult pancreatobiliary reflux in the absence of pancreaticobiliary maljunction. AB - We herein report a case of gallbladder carcinoma associated with occult pancreatobiliary reflux (PR) in the absence of pancreatobiliary maljunction. A 67 year-old woman was referred to our hospital for the evaluation and treatment of a gallbladder tumor. Ultrasonography and computed tomography showed a nodular lesion in the fundus of the gallbladder, indicating the possibility of a gallbladder carcinoma. Endoscopic ultrasonography showed the nodular tumor and thickness of the surrounding epithelium. Endoscopic retrograde cholangiopancreatography revealed a normal pancreaticobiliary junction without the common channel and a slight dilatation of the common bile duct (15 mm in diameter). An open cholecystectomy and partial resection of the liver bed of the gallbladder with regional lymphadenectomy was performed. A C-tube was inserted from the cut end of the cystic duct into the common bile duct to prevent bile stasis. Biliary amylase and lipase levels sampled in the gallbladder were 2604 IU/l and 775 IU/l, respectively. Biliary amylase level in the bile collected from the C-tube in the common bile duct was 119 550 IU/l on postoperative day (POD) 6 and 22 265 IU/l on POD 12. These observations suggested that PR was present in this patient. The histopathological findings of the resected specimen showed a well-differentiated adenocarcinoma of the gallbladder with invasion to the muscle layer and no metastasis of the resected lymph nodes. A high index of nuclear staining for MIB-I in the cancer cells (about 10%) was exhibited, and a few cells in the normal epithelium also stained positive. PMID- 17909728 TI - A phase II trial of doxorubicin and interferon alpha 2b in advanced, non medullary thyroid cancer. AB - BACKGROUND: The combination of doxorubicin and interferon alpha is supported by preclinical data. We sought to evaluate the efficacy of this combination in patients with advanced thyroid cancer. PATIENTS AND METHODS: Patients with locally recurrent or metastatic, radioiodine- refractory thyroid cancer, excluding medullary carcinoma, were treated with interferon alpha-2b 12 million units/m2 subcutaneously on days 1-5 and doxorubicin 40 mg/m2 intravenously, on day 3, every 28 days. RESULTS: 17 patients, 15 with well differentiated and 2 with anaplastic thyroid carcinoma, were enrolled; median age was 69 years. Three patients had received radiation plus low dose doxorubicin previously. In 16 patients assessable for response, 1 patient (6%), who had follicular carcinoma, achieved a partial response and 10 patients (62.5%) stable disease as best response. Median time to progression was 5.9 months and median overall survival 26.4 months. In 14 evaluable patients, 5 (36%) had a thyroglobulin response (30% or more reduction in serum levels). Grade 3/4 neutropenia occurred in 76% of patients and neutropenic fever in 24%. Other grade 3/4 adverse events included fatigue (41%), rigors (18%), fever (6%), nausea/vomiting (29%), anorexia (29%), stomatitis (24%), vision disturbances (18%), neuropathy (18%), and hyponatremia (6%). One patient developed heart failure. CONCLUSIONS: Doxorubicin and interferon alpha was associated with considerable toxicities but modest antitumor activity in patients with advanced, non-medullary thyroid cancer. PMID- 17909729 TI - From evidence based bioethics to evidence based social policies. PMID- 17909730 TI - [Chronic wounds. Novel approaches in research and therapy]. AB - Underlying disease may impair normal wound healing, leading to chronic, poorly healing wounds. Efficient treatment strategies require identification and treatment of the underlying disease as well as directed correction of the wound healing defect. A thorough knowledge of tissue repair mechanisms at the cellular and molecular level will help to achieve these goals. This review focuses on new developments in wound healing research and the resulting non-operative therapeutic implications. PMID- 17909731 TI - [Extraordinarily fast growing tumor in a 1-year-old boy]. PMID- 17909732 TI - [Laser skin resurfacing and fibrin sealing as successful treatment for facial angiofibromas in tuberous sclerosis]. AB - Tuberous sclerosis complex (TSC) is an autosomal-dominant phacomatosis that manifests with visceral harmatomas, epilepsy, and mental retardation. Paranasal angiofibromas may cause bleeding and difficulties in nasal breathing and can stigmatise the individual. When treating TSC patients, the otolaryngologist must take patient compliance and the tendency to develop malignancies into account. We report on a 34-year-old woman who was treated successfully by a combination of CO(2) laser treatment and fibrin glue. The cosmetic results were excellent. PMID- 17909733 TI - [Functional therapy and the limitations for acute elbow dislocation]. AB - Dislocation of the elbow joint is the second most common dislocation, the shoulder being the most common. Non-surgical therapy is done by repositioning and early active motion after a short period of immobilization. There are, however, certain principles which must be followed in order to obtain a favorable result with functional therapy. On the basis of case reports the constraints of early active motion are discussed. Knowing the mechanism or kinematics of an elbow dislocation, it is possible to determine a staging of the injury. Using detailed x-rays and an exact stability test, the degree of instability must be checked after repositioning. It is important to determine the grade of instability and to operatively correct a major instability. PMID- 17909735 TI - [Humeral diaphyseal fractures: functional bracing]. AB - Functional bracing of humeral diaphyseal fractures was conceived after initial experiences with a similar method was used in the management of diaphyseal tibial fractures. Over the years, tibial functional bracing underwent major evolutionary changes, and found its indications basically limited to a smaller group of fractures, consisting of closed, axially unstable fractures that experience at the time of the injury an acceptable degree of shortening, and to transverse fracture that are appropriately reduced and rendered stable. On the other hand, functional bracing of diaphyseal humeral fractures has, maintained the initial indications, contraindications and methodology. This article describes the concept, indications and contraindications of functional bracing of humeral diaphyseal fractures and provides results of 620 fractures with complete follow up. In 97.5% of the patients, the average healing time was 11.5 weeks. 16 patients (2.5%) required operative intervention because of a nonunion and 4 patients (<1 degrees) of the patient had a refracture after brace removal. Nerve function did not return in only one of the 67 patient who had radial nerve palsy. PMID- 17909734 TI - [Cardiac arrest following blunt chest injury. Emergency thoracotomy without ifs or buts?]. AB - In German-speaking countries, most serious thoracic injuries are attributable to the impact of blunt force; they are the second most frequent result of injury after head injury in polytrauma patients with multiple injuries. Almost one in every three polytraumatized patients with significant chest injury develops acute lung failure, and one in every four, acute circulatory failure. The acute circulatory arrest following serious chest injury involves a high mortality rate, and in most cases it reflects a tension pneumothorax, cardiac tamponade, or hemorrhagic shock resulting from injury to the heart or one of the large vessels close to it. Brisk drainage of tension pneumothorax and adequate volume restoration are therefore particularly important in resuscitation of multiply traumatized patients, as are rapid resuscitative thoracotomy to allow direct heart massage, drainage of pericardial tamponade, and control of hemorrhage. However the probability of survival described in the literature is very low for patients sustaining severe chest trauma with acute cardiac arrest. The case report presented here describes a female polytrauma patient who suffered an acute cardiac arrest following cardiac tamponade after admission in the emergency department and who survived without neurological deficits after an emergency thoracotomy. Selections from the topical literature can help the treating physician in the emergency department in making decisions on whether an emergency thoracotomy is indicated after a blunt chest injury and on the procedure itself. PMID- 17909736 TI - [Fractures of the wrist and hand. Principles of conservative treatment]. AB - Over the last years, many special instruments and implants were designed for operative treatment of fractures of the wrist and hand. Therefore, conservative treatment of these injuries became old-fashioned in the eyes of most patients and surgeons. Nevertheless, conservative treatment of nondislocated stable fractures of the wrist and hand usually shows better clinical results than operative treatment does. On the other hand, conservative treatment of displaced fractures is very difficult. The key to success is thorough knowledge of the use of casts, splints, and functional therapy and good patient compliance to avoid complications. Opportunities for conservative treatment of fractures of the wrist and hand are herein described. PMID- 17909737 TI - [Conservative treatment of fractures of the long bones during the growth phase]. AB - In the treatment of fractures of the long bones in children and adolescents, surgical and conservative methods are not mutually exclusive alternatives; rather, each can complement the other in the overall treatment strategy. Many operative procedures, such as fixation of juxtaarticular, metaphyseal fractures, need to be supplemented by conservative procedures, such as immobilization by means of casts. We refer to methods that are performed without anaesthesia and do not involve the implantation of osteosynthetic foreign material as conservative. These are: immobilization with no further treatment, plaster wedging, and functional treatment options. The indications for purely conservative treatment strategies are basically different for articular and nonarticular fractures. Whereas in the case of articular fractures only the nondisplaced fractures are treated by conservative methods, in the case of nonarticular (shaft) fractures those involving a degree of displacement, as long as it is not too pronounced to allow the integration of spontaneous corrections, can also be treated conservatively. PMID- 17909738 TI - [Prostate carcinoma]. AB - Prostate cancer is the most common malignancy in men in Europe, North America, and in some African states. Early diagnosis in an asymptomatic stage is possible through the combination of digitorectal examination, PSA serum testing, and systematic biopsy. However, general screening is so far not recommended by the Urologic Societies, because the efficiency is not yet proved. Imaging is also not recommended for first-line screening. Novel functional methods of transrectal ultrasound (TRUS) and endorectal MRI can improve accuracy of tumor detection to more than 90% and can be used for TRUS- and now also MRI-guided biopsy leading to two- to threefold higher tumor detection rates. There is general agreement that all men over 50 years of age should be informed about the possibilities, benefits, and risks of the available methods for early tumor detection. PMID- 17909739 TI - Analysis of the impact of fibromyalgia on quality of life: associated factors. AB - We analysed the impact of fibromyalgia (FM) on the functional capacity of patients suffering this syndrome and identified factors that are associated with greater disease impact. We performed a cross-sectional descriptive telephone survey on all patients diagnosed with fibromyalgia during 2003 in a university hospital in Spain. Variables studied were socio-demographic, job, clinical, health and psycho-social characteristics of patients diagnosed with FM and impact of FM on them. Disease impact was measured by means of the Fibromyalgia Impact Questionnaire (FIQ). The rest of variables were collected by means of an expressly designed questionnaire. The relation between FIQ score and the other variables was performed with a bivariate analysis, using several tests depending on the variables involved. To analyse the factors associated with greatest disease impact, a multivariate linear regression model was designed. The average FIQ score for the sample was 63.6. Having a larger number of children, being tired and being in a depressed mood were the symptoms that most affected activities of daily living. A diagnosis of any mental illness, reference to repercussion on the family environment, a lower self-rated health and having consulted more specialists before FM diagnosis were associated with a higher impact after adjusting according to all the variables in the model. It can be confirmed that the FIQ is a useful instrument for measuring the impact of FM on quality of life. Identifying factors that determine the extent of its impact will enable more effective therapeutic strategies to be designed. PMID- 17909740 TI - Pregnancy in systemic lupus erythematosus: a retrospective study from a developing community. AB - Little data exists from the developing world on pregnancy in systemic lupus erythematosus (SLE). A 10-year review of pregnancies in lupus patients was conducted at a tertiary hospital in a developing country. Forty-seven pregnancies in 31 patients were identified. Eleven (23%) booked after 20 weeks gestation. There were no maternal deaths; six (13%) mothers experienced flares-all mild. Twelve women developed preeclampsia of which one experienced an intrauterine death. One patient was diagnosed with lupus and nephritis during pregnancy. She required an abortion to control the disease. Another with active nephritis delivered a normal but premature infant despite cyclophosphamide therapy. There was only minor deterioration in renal function. There were 36 (77%) live births, 8 first trimester abortions, 2 elective abortions and 1 still birth. Fourteen (39%) of live births were premature, and five (14%) experienced intrauterine growth retardation (IUGR). Two live-born babies experienced neonatal heartblock, and one, a neonatal lupus rash. We discuss these finding in relation to risk factors and to results from the developed world. PMID- 17909741 TI - Does low-dose and short-term glucocorticoids treatment increase the risk of osteoporosis in rheumatoid arthritis female patients? AB - Rheumatoid arthritis (RA) is frequently complicated by peri-articular and generalized osteoporosis due to increased bone resorption by activated osteoclasts. Pro-inflammatory cytokines, such as TNF-alpha, interleukin 1 (IL1), and interleukin 6 (IL6) are thought, among other factors, to be directly responsible for this extra-articular complication of RA. Glucocorticoids (GCS) commonly prescribed in RA due to their strong anti-inflammatory effect are also well known for causing secondary osteoporosis during a prolonged use. An influence of low-dose GCS therapy (8.7 mg per day) on a bone turnover in female RA patients with or without previous history of GCS treatment was investigated by measuring bone mineral content (BMC), bone mineral density (BMD), and various biochemical markers of inflammation and bone metabolism in comparison to results obtained from: (1) RA patients who have not been treated with GCS and (2) the control group of healthy individuals. Sixty-two female patients with established active RA and 178 healthy individuals from the control group have been investigated. The RA patients were divided into three groups: 21 treated with GCS before the trial--these patients have continued GCS therapy using low doses during the observation; 21 with low-dose GCS therapy launched at the beginning of the trial; and 20 left without GCS treatment. All patients have been assessed twice: at the beginning and after 12 months of observation. BMC and BMD have been measured in all patients in a distal part of forearm. Additionally, several different biochemical markers of osteoporosis and inflammation have been determined. We did not notice any increase in bone metabolism between RA patients receiving GCS therapy for the first time and those treated without GCS after 12 months of observation. Results of BMC, BMD osteocalcin level, total and bone alkaline phosphatase, carboxy-terminal collagen cross links, carboxy-terminal propeptides of type 1 collagen, deoxypyridynoline, and calcium/creatinine ratio were comparable in both groups at the end of the study. There was a significant decrease of the level of IL-6 in patients who had GCS therapy launched at the beginning of observation (p<0.01). However, levels of C-reactive protein (CRP) and alpha1-acid-glycoprotein (AGP) have not changed; the level of ESR dropped significantly (p<0.05) in this group. In contrast, in the group of patients with the previous history of prolonged GCS treatment receiving low doses of GCS during the trial, statistically significant increase of CRP and AGP could be observed (p<0.05) along with further significant worsening of the primary low BMD (p<0.05). Based on the obtained data, we came to the conclusion that anti inflammatory effect of the low-dose GCS therapy in RA patients without previous history of their use may balance their direct negative effect on BMC and BMD. In this group of RA patients, benefits resulting from the 12-month GCS therapy prevail over adverse effects, even if calcium with vitamin D3 supplementation, biphosphonians, or estrogens have not been introduced. On the other hand, low dose GCS therapy could have no benefit for RA patients with the previous history of their prolonged use, as a rise of markers of inflammation and bone turnover, resulting in the further bone loss, has been observed. PMID- 17909742 TI - Formulation of hydrophilic non-aqueous gel: drug stability in different solvents and rheological behavior of gel matrices. AB - PURPOSE: This study was aimed at formulating a hydrophilic non-aqueous gel for topical delivery of the model moisture-sensitive drug, minocycline hydrochloride (MH). METHODS: Stability study of MH dissolved in water and various hydrophilic non-aqueous solvents was performed over a period of four months in order to select a suitable non-aqueous solvent for MH gel. To improve MH stability, the effect of different cation additives on MH stability in the selected solvent was investigated. Non-aqueous gel matrices were prepared from three different types of hydrophilic polymers in glycerin-propylene glycol mixture with Mg(2+) cation additive. Oscillatory shear rheometry was performed on the gel matrices using a cone-and-plate rheometer. RESULTS: MH stability was affected by the type of solvent employed and the duration of storage. Different cation additives affected the extent of MH stabilization through MH-cation complex formation. Rheological properties of the non-aqueous gel matrices were significantly affected by the type and concentration of polymer, and the vehicle ratios in the formulations. CONCLUSIONS: MH stabilization could be achieved using the selected glycerin propylene glycol mixture containing MgCl(2). Gel matrix formulated using this solvent system and 3%w/w N-vinylacetamide/sodium acrylate copolymer had demonstrated the most favorable rheological properties as a gel for topical application. PMID- 17909743 TI - Map-based analysis of genetic loci on chromosome 2D that affect glume tenacity and threshability, components of the free-threshing habit in common wheat (Triticum aestivum L.). AB - During the domestication of bread wheat (Triticum aestivum L.), evolutionary modifications that took place in seed dispersal mechanisms enhanced its suitability for agricultural production. One of these modifications involved the evolution of the free-threshing or hulless characteristic. In this study, we studied quantitative trait loci (QTL) affecting components of the free-threshing habit (threshability and glume tenacity) on chromosome 2D in a recombinant inbred line (RIL) population developed by the International Triticeae Mapping Initiative (ITMI) as well as the tenacious glumes 1 (Tg1) gene in F(2) progeny (CS/CS2D F(2)) of a cross between Chinese Spring and the 2D2 substitution line [Chinese Spring (Ae. tauschii 2D)]. In the ITMI population, two QTL affected threshability (QFt.orst-2D.1 and QFt.orst-2D.2) and their location coincided with QTL affecting glume tenacity (QGt.orst-2D.1 and QGt.orst-2D.2). In the CS/CS2D F(2) population, the location of QTL that affected glume tenacity (QGt.orst-2D.1), the size of a glume base scar after detachment (QGba.orst-2D), and Tg1 (12-cM interval between Xwmc112 and Xbarc168) also coincided. Map comparisons suggest that QFt-orst-2D.1, QGt.orst-2D.1, and QGba.orst-2D correspond to Tg1 whereas QFt.orst-2D.2 and QGt.orst-2D.2 appear to represent separate loci. The observation of coincident QTL for threshability and glume tenacity suggests that threshability is a function of glume adherence. In addition, the observation of the coincident locations of Tg1 and QTL for the force required to detach a glume and the size of a glume base scar after detachment suggests that Tg1's effect on both glume tenacity and threshability resides on its ability to alter the level of physical attachment of glumes to the rachilla of a spikelet. PMID- 17909744 TI - A novel gene, Pi40(t), linked to the DNA markers derived from NBS-LRR motifs confers broad spectrum of blast resistance in rice. AB - Rice blast disease caused by Magnaporthe grisea is a continuous threat to stable rice production worldwide. In a modernized agricultural system, the development of varieties with broad-spectrum and durable resistance to blast disease is essential for increased rice production and sustainability. In this study, a new gene is identified in the introgression line IR65482-4-136-2-2 that has inherited the resistance gene from an EE genome wild Oryza species, O. australiensis (Acc. 100882). Genetic and molecular analysis localized a major resistance gene, Pi40(t), on the short arm of chromosome 6, where four blast resistance genes (Piz, Piz-5, Piz-t, and Pi9) were also identified, flanked by the markers S2539 and RM3330. Through e-Landing, 14 BAC/PAC clones within the 1.81-Mb equivalent virtual contig were identified on Rice Pseudomolecule3. Highly stringent primer sets designed for 6 NBS-LRR motifs located within PAC clone P0649C11 facilitated high-resolution mapping of the new resistance gene, Pi40(t). Following association analysis and detailed haplotyping approaches, a DNA marker, 9871.T7E2b, was identified to be linked to the Pi40(t) gene at the 70 Kb chromosomal region, and differentiated the Pi40(t) gene from the LTH monogenic differential lines possessing genes Piz, Piz-5, Piz-t, and Pi-9. Pi40(t) was validated using the most virulent isolates of Korea as well as the Philippines, suggesting a broad spectrum for the resistance gene. Marker-assisted selection (MAS) and pathotyping of BC progenies having two japonica cultivar genetic backgrounds further supported the potential of the resistance gene in rice breeding. Our study based on new gene identification strategies provides insight into novel genetic resources for blast resistance as well as future studies on cloning and functional analysis of a blast resistance gene useful for rice improvement. PMID- 17909745 TI - The relationship between blood glucose control and intolerance to enteral feeding during critical illness. AB - OBJECTIVE: To assess the relationship between blood glucose concentrations (BSL) and intolerance to gastric feeding in critically ill patients. DESIGN: Prospective, case-controlled study. PATIENTS AND PARTICIPANTS: Two-hourly BSL and insulin requirements over the first 10 days after admission were assessed in 95 consecutive feed-intolerant (NG aspirate > 250 ml during feed) critically ill patients and 50 age-matched, feed-tolerant patients who received feeds for at least 3 days. Patients with diabetes mellitus were excluded. A standard insulin protocol was used to maintain BSL at 5.0-7.9 mmol. MEASUREMENTS AND RESULTS: The peak BSLs were significantly higher before and during enteral feeding in feed intolerant patients. The mean and trough BSLs were, however, similar between the two groups on admission, 24 h prior to feeding and for the first 4 days of feeding. The variations in BSLs over 24 h before and during enteral feeding were significantly greater in feed-intolerant patients. A BSL greater than 10 mmol/l was more prevalent in patients with feed intolerance during enteral feeding. The time taken to develop feed intolerance was inversely related to the admission BSL (r= -0.40). The amount of insulin administered before and during enteral feeding was similar between the two groups. CONCLUSIONS: Feed intolerance in critically ill patients is associated with a greater degree of glycaemic variation, with a greater number of patients with transient hyperglycaemia. These data suggest more intensive insulin therapy may be required to minimize feed intolerance, an issue that warrants further study. PMID- 17909746 TI - Candidemia and candiduria in critically ill patients admitted to intensive care units in France: incidence, molecular diversity, management and outcome. AB - OBJECTIVE: To determine the concomitant incidence, molecular diversity, management and outcome of nosocomial candidemia and candiduria in intensive care unit (ICU) patients in France. DESIGN: A 1-year prospective observational study in 24 adult ICUs. PATIENTS: Two hundred and sixty-two patients with nosocomial candidemia and/or candiduria. MEASUREMENTS AND RESULTS: Blood and urine samples were collected when signs of sepsis were present. Antifungal susceptibility of Candida strains was determined; in addition, all blood and 72% of urine C. albicans isolates were analyzed by using multi-locus sequence type (MLST). The mean incidences of candidemia and candiduria were 6.7 and 27.4/1000 admissions, respectively. Eight percent of candiduric patients developed candidemia with the same species. The mean interval between ICU admission and candidemia was 19.0 +/- 2.9 days, and 17.2 +/- 1.1 days for candiduria. C. albicans and C. glabrata were isolated in 54.2% and 17% of blood and 66.5% and 21.6% of urine Candida-positive cultures, respectively. Fluconazole was the most frequently prescribed agent. In all candidemic patients, the prescribed curative antifungal agent was active in vitro against the responsible identified strain. Crude ICU mortality was 61.8% for candidemic and 31.3% for candiduric patients. Seventy-five percent of the patients were infected with a unique C. albicans strain; cross-transmission between seven patients was suggested in one hospital. CONCLUSIONS: Candidemia is late-onset ICU-acquired infection associated with high mortality. No difference in susceptibility and genetic background were found between blood and urine strains of Candida species. PMID- 17909747 TI - Identification and functional characterisation of cellobiose and lactose transport systems in Lactococcus lactis IL1403. AB - Physiological, biochemical and macroarray analyses of Lactococcus lactis IL1403 and its ccpA and bglR single and double mutants engaged in lactose and beta glucosides catabolism were performed. The kinetic analysis indicated the presence of different transport systems for salicin and cellobiose. The control of salicin catabolism was found to be mediated by the transcriptional regulator BglR and the CcpA protein. The transcriptional analysis by macroarray technology of genes from the PEP:PTS regions showed that several genes, like ybhE, celB, ptcB and ptcA, were expressed at higher levels both in wild type cells exposed to cellobiose and in the ccpA mutant. We also demonstrated that in L. lactis IL1403 cultured on medium with cellobiose and lactose as carbon sources, after the first phase of cellobiose consumption and then co-metabolism of the two sugars, when cellobiose is exhausted the strain uses lactose as the only carbon source. These data could indicate that lactose and cellobiose are transported by a unique system-a PTS carrier induced by the presence of cellobiose, and negatively controlled by the CcpA regulator. PMID- 17909748 TI - Characterisation of colonic accommodation in Wistar Kyoto rats with impaired gastric accommodation. AB - Defective colonic and gastric accommodations have been related to altered viscerosensitivity in irritable bowel syndrome and to functional dyspepsia, respectively. We assessed colonic accommodation in rats with impaired gastric accommodation to determine if altered accommodation can be regarded as a widespread pathophysiological alteration within the gastrointestinal (GI) tract. Colonic accommodation during colorectal distension (CRD) was assessed in Wistar Kyoto rats (WKY), an animal model of impaired gastric accommodation, and in Sprague-Dawley (SD) and Wistar rats, considered normal. CRD (10-80 mmHg)-induced visceral pain responses were also evaluated in the same strains of rats. During gastric distension, WKY rats had lower intra-gastric volume (0.96 +/- 0.22 ml) than SD (1.85 +/- 0.19 ml, P < 0.05) or Wistar rats (2.80 +/- 0.26 ml, P < 0.05), indicating impaired gastric accommodation. In the same animals, pressure-volume curves were constructed during CRD as a measure of colonic accommodation. During short-lasting (1 min) phasic CRD (2-20 mmHg), the pressure-volume curve in WKY rats was displaced to the right compared with SD or Wistar rats, indicative of reduced colonic accommodation (maximal volume: SD, 1.22 +/- 0.05 ml; Wistar, 1.07 +/- 0.04 ml; WKY, 0.87 +/- 0.07 ml; P < 0.01). Pre-treatment with atropine normalised the pressure-volume responses in WKY rats. No differences among strains were observed during the 2-min phasic or ramp-tonic CRD. Visceral pain responses during CRD (10-80 mmHg) were, overall, similar in the three strains, although WKY rats showed lower thresholds for pain (28.0 +/- 4.9 mmHg) than SD (42.3 +/- 6.6 mmHg, P = 0.072) or Wistar rats (48.3 +/- 6.0 mmHg, P < 0.05). WKY rats, although having impaired gastric accommodation, have the ability to fully accommodate the colon to increasing pressures. In WKY rats, impaired accommodation of the smooth muscle might not be a widespread phenomenon along the GI tract but rather a local disturbance. PMID- 17909749 TI - Effects of flavoxate hydrochloride on voltage-dependent Ba2+ currents in human detrusor myocytes at different experimental temperatures. AB - The inhibitory effects of flavoxate hydrochloride (piperidinoethyl-3 methylflavone-8-carboxylate; hereafter referred as flavoxate) on voltage dependent nifedipine-sensitive inward Ba2+ currents (I Ba) in human detrusor myocytes were investigated at different temperatures using conventional whole cell patch-clamp techniques. When the bath-solution temperature was increased from 22 degrees C to 30 degrees C, I Ba peak amplitude was enhanced by approximately twice at several test potentials. Neither the I Ba threshold nor the membrane potentials for the I Ba maximum peak amplitude was affected by the temperature change. The concentration-response curves of flavoxate at both 30 degrees C (Ki = 5.1 microM) and 37 degrees C (Ki = 4.6 microM) were slightly shifted to the left in comparison with that at 22 degrees C (Ki = 10.3 microM). Similar results were also obtained in the presence of nifedipine (Ki = 14 nM at 22 degrees C vs. Ki = 2.5 nM at 30 degrees C and Ki = 2.1 nM at 37 degrees C). Altering the bath-solution temperature from 22 degrees C to 30 degrees C shifted the steady-state inactivation curve of I Ba at -90 mV to the left. At 30 degrees C, the steady-state inactivation curve of I Ba in the presence of flavoxate was also shifted to the left in comparison with that in the absence of flavoxate. Either 3-isobutyl-1-methylxanthine (IBMX) or theophylline, a phosphodiesterase inhibitor, caused little effects on I Ba, although cyclic nucleotides (dibutyryl cAMP and 8-Br-cGMP) inhibited I Ba. These results suggest that the inhibitory actions of flavoxate on I Ba in human detrusor myocytes were slightly changed at different experimental temperatures and that flavoxate directly blocked voltage dependent L-type Ca2+ channels, not through the inhibition of phosphodiesterase activity pathway. PMID- 17909750 TI - Single dose of a dopamine agonist impairs reinforcement learning in humans: behavioral evidence from a laboratory-based measure of reward responsiveness. AB - RATIONALE: The dopaminergic system, particularly D2-like dopamine receptors, has been strongly implicated in reward processing. Animal studies have emphasized the role of phasic dopamine (DA) signaling in reward-related learning, but these processes remain largely unexplored in humans. OBJECTIVES: To evaluate the effect of a single, low dose of a D2/D3 agonist--pramipexole--on reinforcement learning in healthy adults. Based on prior evidence indicating that low doses of DA agonists decrease phasic DA release through autoreceptor stimulation, we hypothesized that 0.5 mg of pramipexole would impair reward learning due to presynaptic mechanisms. MATERIALS AND METHODS: Using a double-blind design, a single 0.5-mg dose of pramipexole or placebo was administered to 32 healthy volunteers, who performed a probabilistic reward task involving a differential reinforcement schedule as well as various control tasks. RESULTS: As hypothesized, response bias toward the more frequently rewarded stimulus was impaired in the pramipexole group, even after adjusting for transient adverse effects. In addition, the pramipexole group showed reaction time and motor speed slowing and increased negative affect; however, when adverse physical side effects were considered, group differences in motor speed and negative affect disappeared. CONCLUSIONS: These findings show that a single low dose of pramipexole impaired the acquisition of reward-related behavior in healthy participants, and they are consistent with prior evidence suggesting that phasic DA signaling is required to reinforce actions leading to reward. The potential implications of the present findings to psychiatric conditions, including depression and impulse control disorders related to addiction, are discussed. PMID- 17909751 TI - Improvement by minocycline of methamphetamine-induced impairment of recognition memory in mice. AB - INTRODUCTION: Cognitive deficits are a core feature of patients with schizophrenia and methamphetamine (METH) psychosis. We have recently found that repeated METH treatment (1 mg/kg, s.c.) in mice, which induces behavioral sensitization, impairs long-term recognition memory in a novel object recognition test (NORT) and that the impairment is ameliorated by clozapine, but not haloperidol. Recent studies indicate that minocycline, a second-generation tetracycline, has potent neuroprotective effects in various animal models of neurological diseases. OBJECTIVES: In the present study, we investigated the effect of minocycline on learning and memory in the NORT and behavioral sensitization in mice that had been administered METH for 7 days. RESULTS: When minocycline (20-40 mg/kg) was administered intraperitoneally once a day for seven consecutive days to mice that had previously been treated with METH for 7 days, it ameliorated the METH-induced impairment of recognition memory in a dose dependent manner, although the same treatment with minocycline had no effect on behavioral sensitization to METH. The administration of minocycline, together with METH, inhibited the development of METH-induced behavioral sensitization. The improvement in memory caused by minocycline was associated with an amelioration of the novelty-induced activation of extracellular signal-regulated kinase 1/2 in the prefrontal cortex of METH-treated mice. CONCLUSIONS: These results suggest that minocycline is useful for the treatment of cognitive deficits in patients with METH psychosis or schizophrenia. PMID- 17909755 TI - Electro-chemiluminescent biosensing. AB - The present review draws a general picture of the bioanalytical applications of electro-chemiluminescent reactions (ECL). Only the two main ECL reactions-i.e. the luminol-based and Ru(bpy)(3)(2+)-based reactions-are considered for application in the fields of enzyme biosensors, immunochemical biosensors, DNA biosensors, and biochips. The mechanism, principle, and experimental conditions of these two reactions are described. Then, for each category of analytical tools, experimental set-ups and performances are presented and discussed. PMID- 17909753 TI - Evaluation of the mu and kappa opioid actions of butorphanol in humans through differential naltrexone blockade. AB - RATIONALE: Butorphanol exerts activity at mu, kappa, and delta opiate receptors in rats and monkeys but produces predominant mu-like effects in humans. OBJECTIVES: The aim of this study was to determine if the kappa receptor-mediated actions of butorphanol could be unmasked or enhanced by giving it in combination with naltrexone, an opioid antagonist with higher affinity for mu vs kappa receptors. MATERIALS AND METHODS: Ten healthy adult inpatient volunteers (eight men, two women), with opioid abuse histories, completed this double-blind, randomized, placebo-controlled study. Naltrexone (0, 1, 3, 10, or 30 mg, p.o.) was administered 1 h before butorphanol (0, 6, or 12 mg/70 kg, i.m.) during 15 test sessions. An array of physiological (e.g., vital signs, urine output, and subject- and observer-rated) measures was collected before and for 4 h after drug administration. RESULTS: Naltrexone alone produced no direct effects. Butorphanol alone produced typical mu-, but not kappa-, related physiological effects (e.g., miosis, respiratory depression) and produced mood and drug effects considered typical of both mu (e.g., "liking," "good drug effects") and kappa agonists (e.g., increases in perceptual disturbances). Naltrexone pretreatment led to significant butorphanol-induced diuresis (i.e., increased urine output and decreased urine osmolality). Naltrexone generally produced a dose-dependent blockade of subjective responses. CONCLUSION: These data suggest that naltrexone antagonism unveiled the kappaergic activity of butorphanol as measured by diuresis, while subjective responses generally attributed to mu vs kappa receptors were not dissociable. Moreover, these data demonstrate that butorphanol exerts physiologically relevant kappa agonist activity at these supraanalgesic doses in humans. PMID- 17909752 TI - Effects of mGlu1 receptor blockade on working memory, time estimation, and impulsivity in rats. AB - RATIONALE: Metabotropic glutamate 1 (mGlu1) receptor antagonists were reported to induce cognitive deficits in several animal models using aversive learning procedures. OBJECTIVE: The present study aimed to further characterize behavioral effects of mGlu1 receptor antagonists using appetitively motivated tasks that evaluate working memory, timing, and impulsivity functions. MATERIALS AND METHODS: Separate groups of adult male Wistar rats were trained to perform four food-reinforced operant tasks: delayed non-matching to position (DNMTP), differential reinforcement of low rates of responding 18 s (DRL 18-s), signal duration discrimination (2-s vs 8-s bisection), and tolerance to delay of reward. Before the tests, rats were pretreated with (3-ethyl-2-methyl-quinolin-6-yl)-(4 methoxy-cyclohexyl)-methanone methanesulfonate (EMQMCM; 2.5-10 mg/kg, i.p.; JNJ16567083). RESULTS: In DNMTP task, EMQMCM produced delay-dependent increases in performance accuracy so that, at 10 mg/kg dose level, percentage of correct lever choices was enhanced at 8- and 16-s delays. In DRL task, at all three tested doses, response rates were higher, and reinforcement rates were lower than under control conditions. In signal duration discrimination tasks, EMQMCM did not have any specific effects on temporal control. In tolerance to delay of reward, EMQMCM (5 and 10 mg/kg) facilitated choice of the lever associated with large reward at longer delay levels. CONCLUSIONS: Blockade of mGlu1 receptors improves working memory and reduces impulsive choice at the doses that have no effects on time perception but appear to facilitate impulsive action. PMID- 17909756 TI - Boronic acid-lectin affinity chromatography. 1. Simultaneous glycoprotein binding with selective or combined elution. AB - We introduce a novel combination of boronic acid affinity chromatography with lectin affinity chromatography, dubbed as boronic acid-lectin affinity chromatography (BLAC). Concanavalin A and wheat germ agglutinin lectins were mixed with the pesudo-lectin boronic acid to form the BLAC affinity column and their performance was evaluated with standard glycoproteins. Optimization of the binding and elution buffers for the BLAC system is described. The BLAC columns were employed to isolate glycoproteins of interest using both selective and/or combined elution. PMID- 17909757 TI - Development of a method for assessing the relative contribution of waterborne and dietary exposure to zinc bioaccumulation in Daphnia magna by using isotopically enriched tracers and ICP-MS detection. AB - In order to study the effect of anthropogenic substances on freshwater and marine ecosystems and to develop methods to derive water-quality criteria, ecotoxicological testing is required. While toxicity assessments are traditionally based on dissolved metal concentrations, assuming that toxicity is caused by waterborne metal only, it was recently pointed out that also the dietary exposure route should be carefully considered and interpreted in regulatory assessments of zinc. In this context, the aim of this experimental study was to develop a method which allows the uptake of waterborne and dietary zinc by Daphnia magna and the interaction between both exposure routes to be studied. Therefore, the setup of a dual isotopic tracer study was required. During several days, daphnids were exposed to 67Zn and 68Zn via the dietary and the waterborne routes, respectively, and after several time intervals the daphnids were sampled and subjected to isotopic analysis by means of inductively coupled plasma mass spectrometry (ICP-MS). In order to obtain reliable and accurate results for zinc, special care was taken to prevent contamination and to deal with the spectral interferences traditionally hindering the determination of zinc. The figures of merit of both a quadrupole-based ICP-MS instrument equipped with a dynamic reaction cell, and a sector field ICP-MS unit were studied, and it was concluded that by using a sector field mass spectrometer operated at medium mass resolution all interferences could be overcome adequately. Although the set up of the exposure experiments seems to be rather simple at first sight, it was shown in this work that several (dynamic) variables can have an influence on the results obtained and on the subsequent data interpretation. The importance of these confounding factors was examined, and on the basis of preliminary calculations it became clear that not only the isotopic composition of the daphnids has to be studied--adequate monitoring of the isotopic composition of the dissolved phase and the algae during the exposure of the daphnids is also required to accurately discriminate between uptake from water and from food. PMID- 17909758 TI - Chemometric experimental design based optimization techniques in capillary electrophoresis: a critical review of modern applications. AB - A critical review of recent developments in the use of chemometric experimental design based optimization techniques in capillary electrophoresis applications is presented. Current advances have led to enhanced separation capabilities of a wide range of analytes in such areas as biological, environmental, food technology, pharmaceutical, and medical analysis. Significant developments in design, detection methodology and applications from the last 5 years (2002-2007) are reported. Furthermore, future perspectives in the use of chemometric methodology in capillary electrophoresis are considered. PMID- 17909759 TI - Elemental mass spectrometry for quantitative proteomics. AB - In the last decade mass-spectrometry-based proteomics has become an indispensable analytical tool for molecular biology, cellular biology and, lately, for the emerging systems biology. This review summarises the evolution and great potential of analytical methods based on elemental mass-spectrometric detection for quantitative proteomic analysis. PMID- 17909760 TI - Analysis of pesticide residues using the Quick Easy Cheap Effective Rugged and Safe (QuEChERS) pesticide multiresidue method in combination with gas and liquid chromatography and tandem mass spectrometric detection. AB - The Quick Easy Cheap Effective Rugged and Safe multiresidue method (QuEChERS) has been validated for the extraction of 80 pesticides belonging to various chemical classes from various types of representative commodities with low lipid contents. A mixture of 38 pesticides amenable to gas chromatography (GC) were quantitatively recovered from spiked lemon, raisins, wheat flour and cucumber, and determined using gas chromatography-tandem mass spectrometry (GC-MS/MS). An additional mixture of 42 pesticides were recovered from oranges, red wine, red grapes, raisins and wheat flour, using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for determination. The pesticides chosen for this study included many of the most frequently detected ones and/or those that are most often found to violate the maximum residue limit (MRL) in food samples, some compounds that have only recently been introduced, as well as a few other miscellaneous compounds. The method employed involved initial extraction in a water/acetonitrile system, an extraction/partitioning step after the addition of salt, and a cleanup step utilizing dispersive solid-phase extraction (D-SPE); this combination ensured that it was a rapid, simple and cost-effective procedure. The spiking levels for the recovery experiments were 0.005, 0.01, 0.02 and 0.2 mg kg(-1) for GC-MS/MS analyses, and 0.01 and 0.1 mg kg(-1) for LC-MS/MS analyses. Adequate pesticide quantification and identity confirmation were attained, even at the lowest concentration levels, considering the high signal-to noise ratios, the very good accuracies and precisions, as well as the good matches between the observed ion ratios. Mean recoveries mostly ranged between 70 and 110% (98% on average), and relative standard deviations (RSD) were generally below 10% (4.3% on average). The use of analyte protectants during GC analysis was demonstrated to provide a good alternative to the use of matrix-matched standards to minimize matrix-effect-related errors. Based on these results, the methodology has been proven to be highly efficient and robust and thus suitable for monitoring the MRL compliance of a wide range of commodity/pesticide combinations. PMID- 17909761 TI - A low perfusion rate microreactor for continuous monitoring of enzyme characteristics: application to glucose oxidase. AB - This report describes a versatile and robust microreactor for bioactive proteins physically immobilized on a polyether sulfone filter. The potential of the reactor is illustrated with glucose oxidase immobilized on a filter with a cut off value of 30 kDa. A flow-injection system was used to deliver the reactants and the device was linked on-line to an electrochemical detector. The microreactor was used for on-line preparation of apoglucose oxidase in strong acid and its subsequent reactivation with flavin adenine dinucleotide. In addition we describe a miniaturized version of the microreactor used to assess several characteristics of femtomole to attomole amounts of glucose oxidase. A low negative potential over the electrodes was used when ferrocene was the mediator in combination with horseradish peroxidase, ensuring the absence of oxidation of electro-active compounds in biological fluids. A low backpressure at very low flow rates is an advantage, which increases the sensitivity. A variety of further applications of the microreactor are suggested. PMID- 17909763 TI - Spectroscopy challenge 12. PMID- 17909762 TI - Rapid genotyping of CYP2D6, CYP2C19 and TPMT polymorphisms by primer extension reaction in a dipstick format. AB - In recent years an increasing amount of interest has been directed at the study and routine testing of polymorphisms responsible for variations in drug metabolism. Most of the current methods involve either time-consuming electrophoresis steps or specialized and expensive equipment. In this context, we have developed a rapid, simple and robust method for genotyping of CYP2D6*3, CYP2D6*4, CYP2C19*2, CYP2C19*3 and TPMT*2 single nucleotide polymorphisms (SNP). Genomic DNA is isolated from whole blood and the segments that span the SNP of interest are amplified by PCR. The products are subjected directly (without purification) to two primer extension (PEXT) reactions (three cycles each) using normal and mutant primers in the presence of biotin-dUTP. The PEXT primers contain a (dA)(30) segment at the 5' end. The PEXT products are detected visually by a dry-reagent dipstick-type assay in which the biotinylated extension products are captured from immobilized streptavidin on the test zone of the strip and detected by hybridization with oligo(dT)-functionalized gold nanoparticles. Patient samples (76 variants in total) were genotyped and the results were fully concordant with those obtained by direct DNA sequencing. PMID- 17909764 TI - Multisensory integration affects ERP components elicited by exogenous cues. AB - Previous studies have shown that the amplitude of event related brain potentials (ERPs) elicited by a combined audiovisual stimulus is larger than the sum of a single auditory and visual stimulus. This enlargement is thought to reflect multisensory integration. Based on these data, it may be hypothesized that the speeding up of responses, due to exogenous orienting effects induced by bimodal cues, exceeds the sum of single unimodal cues. Behavioral data, however, typically revealed no increased orienting effect following bimodal as compared to unimodal cues, which could be due to a failure of multisensory integration of the cues. To examine this possibility, we computed ERPs elicited by both bimodal (audiovisual) and unimodal (either auditory or visual) cues, and determined their exogenous orienting effects on responses to a to-be-discriminated visual target. Interestingly, the posterior P1 component elicited by bimodal cues was larger than the sum of the P1 components elicited by a single auditory and visual cue (i.e., a superadditive effect), but no enhanced orienting effect was found on response speed. The latter result suggests that multisensory integration elicited by our bimodal cues plays no special role for spatial orienting, at least in the present setting. PMID- 17909765 TI - Shift of manual preference by lateralized practice generalizes to related motor tasks. AB - Previous investigation (Teixeira and Teixeira in Brain Cogn, in press, 2007) has evidenced a persistent shift of manual preference for a particular motor task following lateralized practice. In the present study, we assessed the extent to which shift of manual preference is generalizable to related motor tasks. Twenty right-handers were assigned to an experimental or to a control group. The former were provided with practice on a particular sequence of finger movements with their left hand only, while the latter remained inactive. Participants were assessed on manual asymmetry, indexed by movement time, and manual preference for the practiced and for other two sequences of finger movements (transfer tasks). Assessment was made before, immediately after, and 30 days following (retention) practice sessions. Results showed that lateralized practice led to significant bilateral reduction of movement time, maintaining the symmetric performance observed before practice following task acquisition. Regarding manual preference, before task acquisition, all participants in the experimental group were right handed for the main task; immediately after practice their predominant manual preference shifted to the left hand, a profile that was maintained in retention. This persistent shift of manual preference was also observed for one of the transfer tasks requiring the same sequence of transitions between finger movements. Indices of correlation between manual asymmetry and manual preference were non-significant across tasks and phases, suggesting that manual preference was not defined by lateral asymmetry of performance. We propose that manual preference is established by automatic sensorimotor processing and/or increased confidence on a single hand from previous experiences. PMID- 17909766 TI - An object for an action, the same object for other actions: effects on hand shaping. AB - Objects can be grasped in several ways due to their physical properties, the context surrounding the object, and the goal of the grasping agent. The aim of the present study was to investigate whether the prior-to-contact grasping kinematics of the same object vary as a result of different goals of the person grasping it. Subjects were requested to reach toward and grasp a bottle filled with water, and then complete one of the following tasks: (1) Grasp it without performing any subsequent action; (2) Lift and throw it; (3) Pour the water into a container; (4) Place it accurately on a target area; (5) Pass it to another person. We measured the angular excursions at both metacarpal-phalangeal (mcp) and proximal interphalangeal (pip) joints of all digits, and abduction angles of adjacent digit pairs by means of resistive sensors embedded in a glove. The results showed that the presence and the nature of the task to be performed following grasping affect the positioning of the fingers during the reaching phase. We contend that a one-to-one association between a sensory stimulus and a motor response does not capture all the aspects involved in grasping. The theoretical approach within which we frame our discussion considers internal models of anticipatory control which may provide a suitable explanation of our results. PMID- 17909767 TI - Gait deviations induced by visual stimulation in roll. AB - Locomotion control uses proprioceptive, visual, and vestibular signals. The vestibular contribution has been analyzed previously with galvanic vestibular stimulation (GVS), which constitutes mainly a virtual head-fixed rotation in the roll plane that causes polarity-specific deviations of gait. In this study we examined whether a visual disturbance has similar effects on gait when it acts in the same direction as GVS, i.e., when roll vection is induced by head-fixed visual roll motion stimulation. Random dot patterns were constantly rotated in roll at +/-15 degrees /s on a computer-driven binocular head-mounted display that was worn by eight healthy participants. Their gait trajectories were tracked while they walked a distance of 6 m. A stimulation effect was observed only for the first three to four steps, but not for the whole walking distance. These results are similar to the results of previous GVS studies, suggesting that in terms of the direction of action visual motion stimulations in the roll plane are similar to GVS. Both kinds of stimulation cause only initial balance responses in the roll plane but do not contribute to the steering of gait in the yaw plane. PMID- 17909768 TI - Masked priming effect with canonical finger numeral configurations. AB - Discrete numerosities can be represented by various finger configurations. The impact of counting strategies on these configurations and their possible semantic status were investigated in young adults. Experiment 1 showed that young adults named numerical finger configurations faster when they conformed to their own canonical finger-counting habits than when they did not. Experiment 2 showed that numeral finger configurations used as unconsciously presented primes speeded up numerical comparative judgements of Arabic numeral targets. Participants responded faster and made fewer errors with numerical than with non-numerical primes, and when primes and targets were congruent (i.e., leading to the same response). Moreover, this priming effect generalised to novel never consciously seen numerosities for canonical configurations but not for non-canonical ones. These results support the idea that canonical finger configurations automatically activate number semantics whereas non-canonical ones do not. PMID- 17909769 TI - The effect of the "rod-and-frame" illusion on grip planning in a sequential object manipulation task. AB - We investigated the effect of visual context (i.e., a visual illusion) on the planning of a sequential object manipulation task. Participants (n = 13) had to grasp a rod embedded in a "rod-and-frame" illusion and insert the rod-end into a tight hole in a pre-defined way. The grip type (defined by start posture, either pronated or supinated; and end posture, either comfortable or uncomfortable) used to grasp the rod was registered as a macroscopic variable of motor planning. Different rod orientations forced the participants to switch between grip types. As expected, most participants switched between pronated and supinated start postures, such that they ended the movement with a comfortable end posture. As it has been argued that planning is dependent on visual context information, we hypothesized that the visual illusion would affect the specific rod orientation at which participants would switch into a different grip type. This hypothesis was confirmed. More specifically, the illusion affected the critical spatial information that is used for action planning. Collectively, these findings are the first to show an effect of an illusion on motor planning in a sequential object manipulation task. PMID- 17909771 TI - The internal structure of stopping as revealed by a sensory detection task. AB - An important aspect of everyday behaviour is the ability to cancel a prepared movement. In Experiment 1, subjects prepared a response, and then either executed it in response to a subsequent Go signal, or cancelled the movement if a NoGo signal occurred. Subjects had to detect weak shocks, which were delivered after the signals on some trials. Results were compared to a prior instruction condition in which subjects knew at the start of the trial if they should move or not. We found that detection rates on move trials were lower than on non-move trials, consistent with sensory suppression. There was no difference between conditions in detection for move trials. However, detection rates for non-move trials were significantly lower in the NoGo than in the prior instruction condition, suggesting an element of sensory suppression associated with actions, which are prepared, but then inhibited before execution. In Experiment 2, the delay between the NoGo signal and shock was varied. Detection rates improved monotonically as the interval increased from 0 up to 200 ms. The recovery from sensory suppression offers a new way of measuring the processes triggered by a NoGo signal. Our results suggest that when a prepared movement is inhibited the dismantling of the sensory consequences of the motor command takes at least 200 ms. PMID- 17909770 TI - Impaired anticipatory control of force sharing patterns during whole-hand grasping in Parkinson's disease. AB - We examined the coordination of multi-digit grasping forces as they developed during object grasping and lifting. Ten subjects with Parkinson's disease (PD; OFF and ON medication) and ten healthy age-matched control subjects lifted a manipulandum that measured normal forces at each digit and the manipulandum's position. The center of mass (CM) was changed from trial to trial in either a predictable (blocked) or unpredictable (random) order. All subjects modulated individual fingertip forces to counterbalance forces exerted by the thumb and minimize object tilt after lift-off. However, subjects with PD OFF exhibited an impaired ability to use anticipatory mechanisms resulting in less differentiated scaling of individual finger forces to the object CM location. Remarkably, these between-group differences in force modulation dissipated as subjects reached peak grip forces during object lift, although these occurred significantly later in subjects with PD OFF than controls and PD ON. Analysis of the tilt of the object during lift revealed all subjects had similar deviations of the object from the vertical, the direction of which depended on CM location. Thus these findings in subjects with PD indicate that: (a) PD-induced impairments in anticipatory force mechanisms appear to be greatly increased in multi-digit grasping as opposed to previous reports from two-digit grasping; (b) inaccurate scaling of fingertip force amplitude and sharing patterns before object lift is recovered during object lift; (c) the implementation of appropriate force amplitude and sharing among the digits during the lift occurs significantly later than for controls; (d) medication improves the temporal recovery of multi-digit force coordination. These results are discussed within the framework of PD-related deficits in sensorimotor integration and control of multi-degrees of freedom movement. PMID- 17909772 TI - An fMRI study of brain activation in a visual adaptation task: activation limited to sensory guidance. AB - Previous neuroimaging studies yielded different patterns of brain areas activated during sensorimotor adaptation, when sensory conflicts are introduced, e.g. by manipulating visual information. We propose that possible reasons might be the lack to control for adaptation or the change in motor performance. In consequence, it was not possible to distinguish between adaptation-related and error-related brain activations. We have developed a sensorimotor adaptation task which controls for these errors using two types of visual distortion and thus is suited to disambiguate sensorimotor adaptation from the related activation patterns. Twenty healthy subjects were scanned by fMRI during a tracking task, while adapting to a visual distortion, which depended either on hand position or on hand velocity. In either case, adaptation was interleaved with a control condition, designed such that the time-course of tracking errors approximated that under visual distortion. We found that adaptation-related neural activation was limited to the left supramarginal and angular gyrus under the position dependent distortion, but extended bilaterally in the supramarginal gyrus, as well as in the left middle and right superior frontal gyrus under the velocity dependent distortion. Our findings confirm that equating the errors under both conditions will yield an anatomically more restricted activation pattern compared with other studies. The additional recruitment in right parietal and bilateral frontal areas under the velocity-dependent distortion might reflect a higher computational demand, or the involvement of different adaptive mechanisms. PMID- 17909774 TI - How useful are prescribing indicators based on the DU90% method to distinguish the quality of prescribing between pharmacotherapy audit meetings with different levels of functioning? AB - OBJECTIVES: The objective of the study was to assess the association between the quality of drug prescribing based on three indicator types derived from the DU90% method and different levels of functioning in pharmacotherapy audit meetings (PTAMs). MATERIALS AND METHODS: The level of functioning in PTAMs in 2004 was assessed by a standard questionnaire. Data on prescriptions in 2004 by the GPs participating in the included PTAMs were extracted from the database of the Foundation for Pharmaceutical Statistics. Three types of DU90% indicators were computed for the seven mostly prescribed drug classes. With univariate and multivariate analyses of variance, differences in the results of three types of indicators for each of the seven drug classes were assessed according to the levels of PTAMs. RESULTS: For 84 PTAMs with varying levels of functioning, we found no association between the level of PTAM and the quality of prescribing for any of the indicators within the seven drug classes. In general, results gained of all PTAMs seemed to be high in quality for the aspects measured. CONCLUSIONS: It is difficult to define indicators based on the DU90% method, which can readily distinguish differences in the quality of drug prescribing between PTAMs with different levels of functioning. Indicators for prescribing should specifically meet relevant items in the quality of prescribing for certain drug classes. Items for classification of PTAM levels may need some reconsideration. PMID- 17909773 TI - Children with cerebral palsy exhibit greater and more regular postural sway than typically developing children. AB - Following recent advances in the analysis of centre-of-pressure (COP) recordings, we examined the structure of COP trajectories in ten children (nine in the analyses) with cerebral palsy (CP) and nine typically developing (TD) children while standing quietly with eyes open (EO) and eyes closed (EC) and with concurrent visual COP feedback (FB). In particular, we quantified COP trajectories in terms of both the amount and regularity of sway. We hypothesised that: (1) compared to TD children, CP children exhibit a greater amount of sway and more regular sway and (2) concurrent visual feedback (creating an external functional context for postural control, inducing a more external focus of attention) decreases both the amount of sway and sway regularity in TD and CP children alike, while closing the eyes has opposite effects. The data were largely in agreement with both hypotheses. Compared to TD children, the amount of sway tended to be larger in CP children, while sway was more regular. Furthermore, the presence of concurrent visual feedback resulted in less regular sway compared to the EO and EC conditions. This effect was less pronounced in the CP group where posturograms were most regular in the EO condition rather than in the EC condition, as in the control group. Nonetheless, we concluded that CP children might benefit from therapies involving postural tasks with an external functional context for postural control. PMID- 17909775 TI - Pain reduction in osteoporotic patients with vertebral pain without measurable compression. AB - INTRODUCTION: Non-compression osteoporotic vertebral pain (NCOVP) can also cause pain and severe immobilization, such as typical vertebral compression fracture (VCF). The aim of this study was to evaluate whether patients with NCOVP refractory to medical treatment and severely affecting normal daily activities could be offered therapeutic benefit with percutaneous vertebroplasty. METHODS: We conducted a retrospective review of the records of consecutive percutaneous vertebroplasty procedures performed at our institutions during a 28-month period to define a population of patients who suffered from severe NCOVP. Nine such patients were identified based on physical examination, computed tomography, magnetic resonance (MR) imaging, and bone scans. Initial clinical outcomes were assessed by comparing quantitative measurements of pain (10-point scale) and mobility (5-point scale) 1 day before the operation with those 1 day post operation. A second follow-up took place between 2 weeks and 1 month after the operation, with a third follow-up between 6 and 10 months post-operative. Biopsy was taken in each case. RESULTS: Each patient demonstrated point tenderness over radiographically normal-shaped vertebra. Every patient showed a low signal on T1W images, and seven cases showed a high signal on T2W images inside the vertebra, indicating bone marrow edema. All patients experienced a reduction in pain and an increase in mobility after percutaneous vertebroplasty, with a mean pain reduction of 7.0 points and an average improved mobility of 2.8 points. Biopsy results indicated necrotic and/or degenerative changes in eight cases. CONCLUSION: The clinical outcomes of our patients suggest that NCOVP, mainly verified by abnormal MR signals and biopsy results, can be successfully treated by percutaneous vertebroplasty. PMID- 17909776 TI - Clinical applications of diffusion tensor tractography of the spinal cord. AB - Diffusion tensor imaging (DTI) can visualize the white matter tracts in vivo. The aim of this study was to assess the clinical utility of DTI in patients with diseases of the spinal cord. Fourteen subjects underwent magnetic resonance imaging of the spine at 1.5 T. Preliminary diagnosis of the patients suggested traumatic, tumorous, ischemic or inflammatory lesions of the spinal cord. In addition to T2-weighted images, DTI was performed with the gradients in 30 orthogonal directions. Maps of the apparent diffusion coefficient and of fractional anisotropy were reconstructed. Diffusion tensor imaging showed a clear displacement and deformation of the white matter tracts at the level of the pathological lesions in the spinal cord. This capability of diffusion tensor imaging to reliably display secondary alterations to the white matter tracts caused by the primary lesion has the potential to be of great utility for treatment planning and follow-up. PMID- 17909777 TI - Endovascular treatment of basilar and ICA termination aneurysms: effects of the use of HydroCoils on treatment stability in a subgroup of patients prone to a higher recurrence rate. AB - PURPOSE: The aim of this study was to evaluate the stability of occlusion of terminal bifurcation aneurysms after embolization with hydrogel-coated coils. METHODS: Of 35 bifurcation aneurysms, 34 were treated with hydrogel-coated coils in combination with platinum coils, and 1 was treated with hydrogel-coated coils only. Aneurysms were located at the basilar tip in 17 patients, and the internal carotid artery (ICA) bifurcation in 18 patients. The patient population consisted of 20 women and 15 men with ages ranging from 21 to 65 years. The aneurysm was found in 16 patients on presentation for subarachnoid hemorrhage, and in 19 patients the finding was incidental. Of the 35 aneurysms, 25 were small, 9 were large and 1 was giant. The giant aneurysm was located at the basilar tip and showed partial thrombosis. All except two basilar tip aneurysms were treated with balloon assistance. The remaining two basilar tip aneurysms were embolized with the assistance of an aneurysmal neck bridge device. RESULTS: The mean percentage occluded aneurysm volume for all devices was in the range 34-100%. Follow-up angiograms were obtained at 1 year in 6 patients, 2 years in 11 patients, and 3 years in 18 patients. Angiograms obtained immediately after embolization demonstrated a Raymond class 1 occlusion in 29 patients (82.9%) and a Raymond class 2 occlusion in 6 patients (17.1%). In four of these six patients follow-up angiograms demonstrated regrowth with resultant Raymond class 3 occlusion. In the other two patients, Raymond class 2 occlusion remained stable on follow-up angiograms. In patients who had a Raymond class 1 occlusion on the angiogram obtained immediately after embolization, no regrowth was seen on the follow-up angiograms. The overall recanalization rate was 11.4% (three large, one giant) at 6 months. Retreatment was not considered in three of these patients and they were to be followed; the other patient was retreated. CONCLUSION: Our initial procedural data demonstrate that higher volumetric occlusion was achieved with hydrogel-coated coils and the long-term follow-up results showed a favorably low recanalization rates among the terminal bifurcation aneurysms. PMID- 17909779 TI - MR imaging of isolated right subclavian artery. AB - We report the MRI findings in a 3-week-old boy with D-transposition of the great arteries and an abnormal origin of the right subclavian artery from the pulmonary artery. This anomaly of the subclavian arteries is called isolation. It is infrequent in patients with a right aortic arch, but exceedingly rare in those with a left aortic arch. This is a unique report of the MRI findings in this congenital abnormality of the aortic arch. PMID- 17909778 TI - Evaluation of tracheal bronchus in Chinese children using multidetector CT. AB - BACKGROUND: Tracheal bronchus is a congenital bronchial anomaly. The diagnosis should be considered early in intubated patients. Multidetector computed tomography (MDCT) is the newest modality for evaluating tracheal bronchus. OBJECTIVE: To evaluate the utility of 16-slice MDCT in children with tracheal bronchus and to characterize the frequency of tracheal bronchus in children with congenital heart disease. MATERIALS AND METHODS: From June 2005 to May 2007, 3,187 consecutive children (1,124 with congenital heart disease and 2,063 without congenital heart disease) underwent MDCT examination. Minimum-intensity projection reconstruction was performed to show the tracheobronchial tree in every case. RESULTS: Tracheal bronchus was found in 42 children (3.74%) with congenital heart disease but in only 6 children (0.29%) without congenital heart disease. Among the 48 children with tracheal bronchus, 45 had right-side tracheal bronchus and 3 had bilateral tracheal bronchi with heterotaxy syndrome. The diagnostic sensitivity of MDCT was 100% (48/48). CONCLUSION: MDCT is a reliable imaging technique for the diagnosis of tracheal bronchus. Our data showed that right-side tracheal bronchus was more common and bilateral tracheal bronchi usually occurred with heterotaxy syndrome. In addition, tracheal bronchus often occurred with congenital heart disease. The angle between the tracheal bronchus and the trachea is important and should be measured. PMID- 17909781 TI - MRI in the diagnosis of diphallia. AB - Diphallia or duplication of the penis is an extremely rare but well-documented anomaly. According to the presence of one or two corpora cavernosa in each of the penises, diphallia is classified into two major groups of bifid phallus or true diphallia, respectively. We report a 5-year-old boy with duplication of the penis. Significant separation of the penises and their morphology made them appear as true complete phalluses, but MRI established the definitive diagnosis of bifid phallus by demonstrating the presence of one corpus cavernosum in each penis. MRI is a valuable method for achieving the accurate diagnosis of these anomalies and associated malformations. It also provides the appropriate knowledge regarding anatomical detail and assists the surgeon in decision making and preoperative planning for the optimal surgical approach. PMID- 17909780 TI - 123I-Metaiodobenzylguanidine accumulation in a urinoma and cortex of an obstructed kidney after surgical resection of an abdominal neuroblastoma. AB - Surgical ureteric injury is rare and often unsuspected for a long time. We present a child in whom an abdominal neuroblastoma was completely excised, but during surgery the left ureter was transected and anastomosed. One month later, during postoperative disease staging, abnormal (123)I-MIBG accumulation was observed in the left renal cortex and the left side of the abdomen. These findings were consistent with acute total obstruction and urinoma formation and were subsequently confirmed by renography and MRI. Despite treatment efforts, a significant amount of left renal mass and function were lost over the following months. These unusual findings are new additions to the literature regarding potential false-positive interpretations of (123)I-MIBG scans. PMID- 17909782 TI - Fractional anisotropy in white matter tracts of very-low-birth-weight infants. AB - BACKGROUND: Advances in neonatal intensive care have not yet reduced the high incidence of neurodevelopmental disability among very-low-birth-weight (VLBW) infants. As neurological deficits are related to white-matter injury, early detection is important. Diffusion tensor imaging (DTI) could be an excellent tool for assessment of white-matter injury. OBJECTIVE: To provide DTI fractional anisotropy (FA) reference values for white-matter tracts of VLBW infants for clinical use. MATERIALS AND METHODS: We retrospectively analysed DTI images of 28 VLBW infants (26-32 weeks gestational age) without evidence of white-matter abnormalities on conventional MRI sequences, and normal developmental outcome (assessed at age 1-3 years). For DTI an echoplanar sequence with diffusion gradient (b = 1,000 s/mm(2)) applied in 25 non-collinear directions was used. We measured FA and apparent diffusion coefficient (ADC) of different white-matter tracts in the first 4 days of life. RESULTS: A statistically significant correlation was found between gestational age and FA of the posterior limb of the internal capsule in VLBW infants (r = 0.495, P<0.01). CONCLUSION: Values of FA and ADC were measured in white-matter tracts of VLBW infants. FA of the pyramidal tracts measured in the first few days after birth is related to gestational age. PMID- 17909783 TI - An unusual case of left aberrant innominate artery with right aortic arch: evaluation with high-resolution CT. AB - A left aberrant innominate (brachiocephalic) artery is an angiographically well known entity that may cause tracheal compression. We report a male newborn who was admitted for further investigation of a prenatally suspected major vessel anomaly. High-resolution CT was used to completely assess the abnormal anatomy and the relationship with the airway, as well as to guide the surgical approach for its correction. PMID- 17909784 TI - Hypervitaminosis A-induced premature closure of epiphyses (physeal obliteration) in humans and calves (hyena disease): a historical review of the human and veterinary literature. AB - Vitamin A toxicity in the infant, which now occurs rarely from dietary overdosage, was recognized in the 1940s as painful periostitis with rare progression to premature closure of the lower limb epiphyses. Decades later, most cases of vitamin A-induced premature epiphyseal closure (physeal obliteration) occur in pediatric dermatologic patients given vitamin A analogues. This phenomenon resembles a strange disease discovered in more recent years in calves with closed epiphyses of the hind limbs, known as hyena disease. This was a mystery until proved to be caused by vitamin A toxicity from enriched grain that causes the calves to have short hind limbs that resemble those of a hyena and gait disturbance. This historical review links the human and veterinary literature in terms of vitamin A-induced epiphyseal closure using a case report format of a 16-month-old human infant with closed knee epiphyses and gait disturbance that is reminiscent of hyena disease seen in calves. PMID- 17909785 TI - Analyses of the acetate-producing pathways in Corynebacterium glutamicum under oxygen-deprived conditions. AB - Corynebacterium glutamicum R efficiently produces valuable chemicals from glucose under oxygen-deprived conditions. In an effort to reduce acetate as a byproduct, acetate productivity of several mutant-disrupted genes encoding possible key enzymes for acetate formation was determined. Disruption of the aceE gene that encodes the E1 enzyme of the pyruvate dehydrogenase complex resulted in almost complete elimination of acetate formation under oxygen-deprived conditions, implying that acetate synthesis under these conditions was essentially via acetyl coenzyme A (CoA). Simultaneous disruption of pta, encoding phosphotransacetylase, and ack, encoding acetate kinase, resulted in no measurable change in acetate productivity. A mutant strain with disruptions in pta, ack and as-yet uncharacterized gene (cgR2472) exhibited 65% reduced acetate productivity compared to the parental strain, although a single disruption of cgR2472 exhibited no effect on acetate productivity. The gene cgR2472 was shown to encode a CoA-transferase (CTF) that catalyzes the formation of acetate from acetyl-CoA. These results indicate that PTA-ACK as well as CTF is involved in acetate production in C. glutamicum. This study provided basic information to reduce acetate production under oxygen-deprived conditions. PMID- 17909786 TI - Kinetics of consumption of fermentation products by anode-respiring bacteria. AB - We determined the kinetic response of a community of anode-respiring bacteria oxidizing a mixture of the most common fermentation products: acetate, butyrate, propionate, ethanol, and hydrogen. We acclimated the community by performing three consecutive batch experiments in a microbial electrolytic cell (MEC) containing a mixture of the fermentation products. During the consecutive-batch experiments, the coulombic efficiency and start-up period improved with each step. We used the acclimated biofilm to start continuous experiments in an MEC, in which we controlled the anode potential using a potentiostat. During the continuous experiments, we tested each individual substrate at a range of anode potentials and substrate concentrations. Our results show low current densities for butyrate and hydrogen, but high current densities for propionate, acetate, and ethanol (maximum values are 1.6, 9.0, and 8.2 A/m(2), respectively). Acetate showed a high coulombic efficiency (86%) compared to ethanol and propionate (49 and 41%, respectively). High methane concentrations inside the MEC during ethanol experiments suggest that methanogenesis is one reason why the coulombic efficiency was lower than that of acetate. Our results provide kinetic parameters, such as the anode overpotential, the maximum current density, and the Monod half-saturation constant, that are needed for model development when using a mixture of fermentation products. When we provided no electron donor, we measured current due to endogenous decay of biomass (approximately 0.07 A/m(2)) and an open-cell potential (-0.54 V vs Ag/AgCl) associated with biomass components active in endogenous respiration. PMID- 17909787 TI - Improving PCR and qPCR detection of hydrogenase A (hydA) associated with Clostridia in pure cultures and environmental sludges using bovine serum albumin. AB - Detection of hydA genes of Clostridia spp. using degenerative and species specific primers for C. butyricum were optimized by the addition of bovine serum albumin (BSA) to polymerase chain reaction (PCR) and quantitative PCR (qPCR) reactions. BSA concentrations ranging from 100 to 400 ng/microl were examined using pure cultures and a variety of environmental samples as test targets. A BSA concentration of 100 ng/microl, which is lower than previously reported in the literature, was found to be most effective in improving the detection limit. The brightness of amplicons with 100 ng/mul BSA increased in ethidium bromide-treated gels, the minimum detection limit with BSA was at least one log greater, and cycle threshold (C(T)) values were lower than without BSA in qPCR indicating improved detection of target deoxyribonucleic acid for most samples tested. Although amplicon visualization was improved at BSA concentrations greater than or equal to 100 ng/microl, gene copy numbers detected by qPCR were less, C(T) values were increased, and T(m) values were altered. SYBR Green dissociation curves of qPCR products of DNA from pure culture or sludge samples showed that BSA at 100 ng/microl reduced the variability of peak areas and T(m) values. PMID- 17909788 TI - Oleic acid delays and modulates the transition from respiratory to fermentative metabolism in Saccharomyces cerevisiae after exposure to glucose excess. AB - This work aimed to study the transition from respiratory to fermentative metabolism in Saccharomyces cerevisiae CEN.PK 113-7D and more specifically to evaluate the implication of the acetyl-coenzymeA-derived carbon transport from cytosol to mitochondria in the onset of the metabolic shift. The strategy consisted in introducing, during aerobic glucose-limited chemostat (D = 0.16 h( 1)), [corrected] a local perturbation around the step to be studied by the addition of cosubstrate and in analyzing the consequences of such a perturbation on the metabolic transition. Oleic acid and L: -carnitine were among the tested cosubstrates because they were known to stimulate enzymes implicated in the acetyl-coenzymeA transport between the different cell compartments, such as the carnitine acetyl transferases. The metabolic transition was then comparatively quantified in sole glucose and in glucose/oleic acid chemostats in presence/absence of L: -carnitine after a pulse of glucose. Feeding the culture with oleic acid (D (ole) = 0.0041 and 0.0073 h(-1)) [corrected] led to a delay in the onset of the metabolic shift (up to 15 min), a 33% decrease in the ethanol production and a redirection of the carbon flux toward biomass production. The data clearly showed a modulation of the carbon distribution among respiration and fermentation, in favor of a decrease in the "short-term" Crabtree effect by the oleic acid. PMID- 17909789 TI - Rubor, calor, tumor, dolor, functio laesa... or molecular imaging. PMID- 17909790 TI - Comparison of (18)F-FLT PET and (18)F-FDG PET for preoperative staging in non small cell lung cancer. AB - PURPOSE: The nucleoside analog 3'-deoxy-3'-(18)F-fluorothymidine (FLT) has been introduced for imaging cell proliferation with positron emission tomography (PET). We prospectively compared the diagnostic efficacy of FLT PET with that of 2-deoxy-2-(18)F-fluoro-D-glucose (FDG) PET for the preoperative nodal and distant metastatic staging of non-small cell lung cancer (NSCLC). METHODS: A total of 34 patients with NSCLC underwent FLT PET and FDG PET. PET imaging was performed at 60 min after each radiotracer injection. The PET images were evaluated qualitatively for regions of focally increased metabolism. For visualized primary tumors, the maximum standardized uptake value (SUV) was calculated. Nodal stages were determined by using the American Joint Committee on Cancer staging system and surgical and histologic findings reference standards. RESULTS: For the depiction of primary tumor, sensitivity of FLT PET was 67%, compared with 94% for FDG PET (P = 0.005). Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy for lymph node staging on a per-patient basis were 57, 93, 67, 89, and 85%, respectively, with FLT PET and 57, 78, 36, 91, and 74%, respectively, with FDG PET (P > 0.1 for all comparisons). Two of the three distant metastases were detected with FLT and FDG PET. CONCLUSION: In NSCLC, FLT PET showed better (although not statistically significant) specificity, positive predictive value and accuracy for N staging on a per patient basis than FDG PET. However, FDG PET was found to have higher sensitivity for depiction of primary tumor than FLT PET. PMID- 17909791 TI - CT-based attenuation correction in (82)Rb-myocardial perfusion PET-CT: incidence of misalignment and effect on regional tracer distribution. AB - PURPOSE: Misalignment of low-dose-CT used for attenuation correction (AC) may cause artifacts in cardiac-PET-CT. The aim was to evaluate incidence and severity of misalignment and its quantitative effects on regional myocardial (82)Rb distribution. METHODS: Rest/dipyridamole (82)Rb-perfusion-PET-CT studies of 92 consecutive patients were analyzed for misalignment. Two different scanning protocols were employed: the first 57 patients had separate CTs for rest and stress PET. The following 35 patients had one CT at rest, used for AC of rest and stress PET. Misalignment was visually scored on a five-point scale (0 = no, 1 = minimal, 2 = mild, 3 = moderate, and 4 = severe). In five representative patients with normal perfusion and low probability of disease, 95 polarmaps were created by shifting CT vs PET prior to reconstruction of attenuation-corrected data sets using dedicated software (three dimensions of space; magnitude of shifts, 5, 10, 14 mm). RESULTS: PET/CT -misalignment was detected in 60% of rest and 67% of stress studies. Alignment for rest was better than that for stress (0.7 +/- 0.7 vs 1.0 +/- 0.9, P = 0.03). Comparison of the two protocols revealed no effect on the alignment of the stress study (1.0 +/- 0.9 vs 1.0 +/- 0.9, P = 0.9). Quantitatively, the largest individual effect of any artificial misalignment was a 25% reduction of relative (82)Rb uptake. With a shift of 1 cm, the largest effect in an individual was a 19% decrease. Anterior wall was most frequently influenced by misalignment, but changes of uptake also occurred in all other segments. CONCLUSIONS: Misalignment between CT and PET in cardiac-PET-CT influences regional tracer distribution in multiple segments. Repeated CT imaging after dipyridamole does not improve alignment. These results emphasize the need for strategies to improve coregistration in clinical imaging protocols. PMID- 17909793 TI - Semiautomatic volume of interest drawing for (18)F-FDG image analysis-method and preliminary results. AB - PURPOSE: Functional imaging of cancer adds important information to the conventional measurements in monitoring response. Serial (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET), which indicates changes in glucose metabolism in tumours, shows great promise for this. However, there is a need for a method to quantitate alterations in uptake of FDG, which accounts for changes in tumour volume and intensity of FDG uptake. Selection of regions or volumes [ROI or volumes of interest (VOI)] by hand drawing, or simple thresholding, suffers from operator-dependent drawbacks. MATERIALS AND METHODS: We present a simple, robust VOI growing method for this application. The method requires a single seed point within the visualised tumour and another in relevant normal tissue. The drawn tumour VOI is insensitive to the operator inconsistency and is, thus, a suitable basis for comparative measurements. The method is validated using a software phantom. We demonstrate the use of the method in the assessment of tumour response in 31 patients receiving chemotherapy for various carcinomas. RESULTS: Valid assessment of tumour response could be made 2-4 weeks after starting chemotherapy, giving information for clinical decision making which would otherwise have taken 9-12 weeks. Survival was predicted from FDG-PET 2-4 weeks after starting chemotherapy (p = 0.04) and after 9-12 weeks FDG-PET gave a better prediction of survival (p = 0.002) than CT or MRI (p = 0.015). CONCLUSIONS: FDG-PET using this method of analysis has potential as a routine tool for optimising use of chemotherapy and improving its cost effectiveness. It also has potential for increasing the accuracy of response assessment in clinical trials of novel therapies. PMID- 17909792 TI - Pretargeting CWR22 prostate tumor in mice with MORF-B72.3 antibody and radiolabeled cMORF. AB - PURPOSE: We have now applied our MORF/cMORF pretargeting technology to the targeting of CWR22 prostate tumor in nude mice. METHODS: The antiTAG-72 antibody B72.3 was conjugated with an 18 mer MORF while the cMORF was radiolabeled with (99m)Tc. The specific binding of the antibody to the CWR22 cells was first confirmed in an assay placing the radiolabeled B72.3 antibody in competition with increasing concentrations of native B72.3. Thereafter, a group of four CWR22 tumored mice intravenously received the MORF-B72.3 and, 3 days later, the (99m)Tc cMORF, and were killed at 3 h postradioactivity injection. The dosage of the labeled cMORF was selected on the basis of previous experience in LS174T tumored mice. As controls, four animals received only the radiolabeled cMORF and another four received only the (111)In-B72.3. The maximum percent tumor accumulation (MPTA) of the labeled cMORF was subsequently determined by a dosage study of labeled cMORF. Both a multipinhole SPECT image and a planar gamma camera image were obtained of a representative mouse. RESULTS: The CWR22 tumor was confirmed to be TAG-72-positive. The MPTA of the labeled cMORF in the CWR22 tumor was 2.22%ID/g compared to only 0.12%ID/g in control mice without pretargeting. Both the planar and tomographic images confirmed the success of the CWR22 pretargeting. CONCLUSIONS: The MORF/cMORF pretargeting approach has been successfully applied to tumor targeting of the prostate xenograft CWR22. However, the MPTA in this tumor model is lower than that in the LS174T tumor model investigated earlier, possibly due to a lower tumor blood supply. PMID- 17909794 TI - Imaging the norepinephrine transporter with positron emission tomography: initial human studies with (S,S)-[18F]FMeNER-D2. AB - INTRODUCTION: (S,S)-[(18)F]FMeNER-D(2) is a recently developed positron emission tomography (PET) ligand for in vivo quantification of norepinephrine transporter. A monkey occupancy study with the radioligand indicated that (S,S)-[(18)F]FMeNER D(2) can be useful for quantitative PET analysis. In this preliminary study, regional distributions in the living human brain were evaluated. MATERIALS AND METHODS: Brain PET measurements were performed for a total of 255 min after the injection of 188.3 +/- 5.7 MBq of (S,S)-[(18)F]FMeNER-D(2) in four healthy male subjects. Regions of interests were drawn on the thalamus and the caudate in the coregistered MRI/PET images. RESULTS: (S,S)-[(18)F]FMeNER-D(2) displayed good brain penetration and selective retention in regions rich in norepinephrine reuptake sites. The transient peak equilibrium was reached during the PET measurements. The ratios of radioactivity uptake in the thalamus to that in the caudate were 1.50 +/- 0.06 for the time period of 90-255 min. CONCLUSION: The present preliminary investigation indicates that (S,S)-[(18)F]FMeNER-D(2) has suitable characteristics for probing the norepinephrine reuptake system with PET in the human brain. PMID- 17909795 TI - Scalene muscle uptake: a potential pitfall in head and neck PET/CT. AB - PURPOSE: To describe increased 2-deoxy-2-[(18)F] fluoro-D-glucose (FDG) uptake in the scalene muscles in a large population of patients referred for evaluation with FDG positron emission tomography/computed tomography (PET/CT) imaging. METHODS: The study met criteria for institutional review board exemption. FDG PET/CT images from 410 patients (179 males; mean age 56.8 years, range 6-88) were retrospectively reviewed for the presence or absence of FDG uptake in the neck that corresponded to the scalene muscles on the concurrent CT scan. Medical records were reviewed and data including age, sex, smoking history, reason for referral, and history of obstructive airways disease, thoracotomy, and thoracic radiation were recorded and evaluated. RESULTS: One hundred and forty-seven of the 410 scans (36%) demonstrated increased FDG uptake on PET that corresponded to the scalene muscles on the CT scan. The uptake was most often bilateral, symmetrical, and linear (n = 117). Other patterns of scalene muscle uptake included unilateral and linear uptake (n = 27) and unilateral and focal uptake (n = 3). Scalene muscle uptake was more common in patients referred for evaluation of lung carcinomas compared to other types of tumors (52% vs. 32%, p = 0.05). CONCLUSION: Linear FDG uptake in scalene muscles is a commonly seen pattern on PET/CT. This finding should be recognized as a distinct entity and not misinterpreted on transverse images as metastatic disease. PMID- 17909796 TI - The effect of catecholamines on the glucose uptake in brown adipose tissue demonstrated by (18)F-FDG PET/CT in a patient with adrenal pheochromocytoma. PMID- 17909797 TI - Impact of the CCR5 gene polymorphism on the survival of metastatic melanoma patients receiving immunotherapy. AB - PURPOSE: Chemokines influence both tumor progression and anti-tumor immune response. A 32-bp-deletion polymorphism in the chemokine receptor 5 gene (CCR5Delta32) has been shown to result in a non-functional protein. This study was aimed at evaluating the potential impact of this gene polymorphism on disease progression and treatment outcome in patients with melanoma. PATIENTS AND METHODS: CCR5 genotyping was performed by PCR on DNA extracted from serum samples of 782 cutaneous melanoma patients with known disease history and long-term clinical follow-up. Genotypes were correlated with patient survival and types of treatment. RESULTS: Of 782 melanoma patients, 90 (11.5%) were heterozygous and 12 (1.5%) were homozygous for CCR5Delta32. Analyzing the complete cohort, the disease-specific survival from date of primary diagnosis was not influenced by CCR5 status. Similarly, no significant impact could be detected on the treatment outcome of stage III patients. In 139 stage IV patients receiving immunotherapy, CCR5Delta32 was associated with a decreased survival compared to patients not carrying the deletion (median 12.5 vs. 20.3 months, P = 0.029). Multivariate analysis revealed the CCR5 genotype as an independent factor impacting disease specific survival in this patient population (P = 0.002), followed by gender (P = 0.019) and pathological classification of the primary (pT; P = 0.022). CONCLUSION: The presence of the CCR5Delta32 polymorphism in patients with stage IV melanoma results in a decreased survival following immunotherapy and may help to select patients less likely to benefit from this type of treatment. PMID- 17909798 TI - Hydatid cyst mimicking acute chest syndrome in a sickle thalassemia patient. PMID- 17909800 TI - Primary plasma cell leukemia with initial cutaneous involvement and IgA biclonal gammopathy. PMID- 17909799 TI - High expression of CEACAM6 and CEACAM8 mRNA in acute lymphoblastic leukemias. AB - CEACAM family members are a set of widely expressed proteins involved in several biological functions, including cell adhesion, migration, signal transduction, and the regulation of gene expression. Abnormal overexpression and downregulation of some CEACAMs have been described in tumor cells. Monoclonal antibodies grouped in the CD66 cluster recognize CEACAM members. Ectopic CD66 expression is commonly detected in B-cell lineage acute lymphoblastic leukemia (ALL). To investigate the CEACAM messenger RNA (RNA) expression in leukemic blasts, we performed a quantitative polymerase chain reaction (RQ-PCR) analysis in purified RNA samples from a consecutive series of acute leukemias (135 patients). Most B-cell lineage ALL expressed CD66 (79.5%), whereas no single case of T-cell lineage ALL disclosed CD66 reactivity (0%). All the BCR-ABL+ ALL cases showed CD66 expression. CD66 was positive even in cases without CD10 expression (72.7%) and/or with MLL rearrangements. Despite the sharp contrast between T-ALL and B ALL in CD66 reactivity, CEACAM patterns were comparable, and only minor differences for CEACAM1 and CEACAM8 were detected. All the leukemic samples showed overexpression of CEACAM6 and 8 when compared with normal granulocytes. These results were confirmed by dilutional experiments. The leukemic pattern paralleled the normal regenerating bone marrow with lower values for CEACAM1. In line with the results for CD66 reactivity, neoplastic cell lines had a uniform low expression of CEACAM family members. It remains to be investigated whether these CEACAM disturbances provide growth advantages to tumoral cells by inhibiting the anoikis process. PMID- 17909801 TI - Identification of two distinct populations of endothelial progenitor cells differing in size and antigen expression from human umbilical cord blood. AB - Endothelial progenitor cells (EPCs) have been isolated from peripheral blood, bone marrow, and umbilical cord blood (CB) and determined to be in heterogeneous populations; however, specific variations in their characteristics remain to be clarified. In this study, we observed that mononuclear cells (MNCs) of CB change in morphology to differentiate into mature endothelial cells (EC) after 6 weeks of culture. In early days of culture along with the differentiation, two distinct populations of EPCs were detected, defined by two-dimensional dot plots (forward scatter vs side scatter) with flow cytometry, namely, relatively small cells (S EPCs) and relatively large cells (L-EPCs). S-EPCs were found to express CD34 but not CD14, while the converse was the case for L-EPCs. When CD34(+)/CD14(-) cells and CD34(-)/CD14(+) cells were isolated from original MNCs of CB and cultured independently, S-EPCs and L-EPCs were derived from CD34(+)/CD14(-) and from CD34( )/CD14(+) cells, respectively. Furthermore, when the two EPCs at day 7 were separated by cell sorter and recultured, there was no crossover in terms of CD34 and CD14 expression. While expression of VE-cadherin and vascular endothelial growth factor receptor-2 (VEGFR-2) on L-EPCs was significantly greater than on S EPCs, levels of CD31 were lower. In addition, L-EPCs exhibited greater proliferative ability on stimulation with VEGF. Although these two EPCs expressed different phenotypes, including growth factor receptors, and had different proliferative ability, they both eventually differentiated into mature ECs after more than 3 weeks of culture. PMID- 17909802 TI - Consumption of silibinin, a flavonolignan from milk thistle, and mammary cancer development in the C3(1) SV40 T,t antigen transgenic multiple mammary adenocarcinoma (TAg) mouse. AB - Silibinin is a flavonolignan extracted from milk thistle with cancer chemopreventive activity in preclinical models of prostate and colorectal cancer. A milk thistle extract, of which silibin is a major component, has recently been shown to exacerbate mammary carcinogenesis in two rodent models. We tested the hypothesis that consumption of silibinin or silipide, a silibinin formulation with pharmaceutical properties superior to the unformulated agent, affect breast cancer development in the C3(1) SV40 T,t antigen transgenic multiple mammary adenocarcinoma mouse model. Mice received silibinin or silipide (0.2% silibinin equivalents) with their diet from weaning, and tumour development was monitored by weekly palpation and the number and weight of neoplasms at the end of the experiment. Intervention neither promoted, nor interfered with, tumour development. The result suggests that promotion of carcinogenesis is not a feature of silibinin consistent across rodent models of mammary carcinogenesis. PMID- 17909803 TI - Temozolomide in combination with fotemustine in patients with metastatic melanoma. AB - PURPOSE: Temozolomide and fotemustine are both active drugs for treating metastatic melanoma. The present study was designed to assess the efficacy and safety of combination therapy with temozolomide + fotemustine in patients with metastatic melanoma. METHODS: Forty patients (median age 50.5 and 22 males) with pathologically confirmed, unresectable, AJCO stage IV melanoma were enrolled into the study. The primary endpoints were tumor response and safety. Patients received oral temozolomide 125 mg/m(2) on days 1-7 and intravenous fotemustine 80 mg/m(2) on day 3 every 3 weeks. RESULTS: Fourteen (35%) patients achieved an objective response, including 3 (7.5%) complete and 11 (27.5%) partial responses. Median overall survival time was 6.7 months and 6-month survival rate was 57.4%. Myelosupression, particularly thrombocytopenia, was the primary toxicity. CONCLUSION: The regimen, temozolomide combined with fotemustine, is an active and moderately safe first-line chemotherapy regimen with acceptable and easily manageable toxicities in patients with metastatic melanoma. PMID- 17909804 TI - Phase I clinical trial of intrathecal gemcitabine in patients with neoplastic meningitis. AB - PURPOSE: A phase I study of intrathecal (IT) gemcitabine was performed to define a safe dose and characterize the toxicity profile and CSF pharmacokinetics of gemcitabine and its major metabolite 2',2'-difluoro-deoxyuridine (dFdU) in patients 3 years of age and older with neoplastic meningitis. EXPERIMENTAL DESIGN: Gemcitabine was administered via Ommaya reservoir or lumbar puncture at three dose levels: 5 mg weekly, 5 mg twice-weekly, and 10 mg twice-weekly using a standard phase I dose escalation design. Serial CSF samples were obtained for pharmacokinetic studies in seven patients with Ommaya reservoirs. Serial blood samples for pharmacokinetic studies were also obtained from three patients. RESULTS: Ten patients were enrolled in this study. Significant neurological toxicities occurred in two patients including myelitis in a patient at the 5 mg twice-weekly dose level and somnolence in a patient at the 10 mg twice-weekly dose level. No complete responses were seen; however, three patients had stable disease. Gemcitabine was rapidly eliminated from the CSF with a terminal half life of 61 +/- 50 min. No gemcitabine or dFdU was detected in plasma. CONCLUSIONS: IT gemcitabine was associated with significant neurotoxicity; therefore, its further development for IT use is not recommended. PMID- 17909805 TI - A phase I study of docetaxel with ketoconazole modulation in patients with advanced cancers. AB - PURPOSE: The aims were to determine the maximum tolerable dose (MTD) of docetaxel with CYP3A inhibition by ketoconazole, and to correlate the pharmacokinetics of docetaxel with midazolam phenotyping of CYP3A activity. METHODS: Forty-one patients with refractory metastatic cancers were treated with an escalating dose of intravenous docetaxel once in every 3 week of 10 mg/m(2), concurrently with oral ketoconazole 200 mg twice daily for 3 days starting 2 days before the administration of docetaxel. Midazolam phenotyping test with ketoconazole modulation was performed before the first cycle of docetaxel. Docetaxel and midazolam pharmacokinetics were compared to our previous study of docetaxel treatment without ketoconazole modulation. RESULTS: Neutropenia was the dose limiting toxicity. The maximum tolerated dose was 70 mg with mean AUC at 70 mg similar to 75 mg/m(2) of docetaxel without ketoconazole. The plasma clearances of docetaxel and midazolam were reduced by 1.7- and 6-fold, respectively. The variability of midazolam AUC was reduced from 157 to 67%, but variability of docetaxel clearance was not reduced by CYP3A inhibition. Docetaxel clearance correlated with renal function and maximum concentration of ketoconazole, but not midazolam clearance or other variables of hepatic function. CONCLUSION: Fixed dosing was found to be feasible, without increased variability of clearance or neutrophil toxicity compared to BSA-based dosing. With ketoconazole modulation, docetaxel clearance correlated with renal function but not CYP3A phenotype. PMID- 17909806 TI - Effect of N-acetylcysteine route of administration on chemoprotection against cisplatin-induced toxicity in rat models. AB - Dosing and route of administration of N-acetylcysteine (NAC) for protection against cisplatin (CDDP) nephrotoxicity was investigated in rats. Two models of toxicity were tested: a single high dose of CDDP (10 mg/kg intraperitoneally (IP)), and multiple low dose treatments (1 mg/kg IP twice a day for 4 days, 10 days rest, then repeated). NAC (50-1,200 mg/kg) was given to the rats by IP, oral (PO), intravenous (IV) and intra-arterial (IA) routes. Renal toxicity was determined by blood urea nitrogen (BUN) and creatinine (CR) levels 3 days after treatment. Blood collected 15 min after NAC was analyzed for total NAC. Both models of CDDP administration produced renal toxicity. In the single dose CDDP model, NAC 400 mg/kg given IP and PO produced no renal protection as measured by BUN (131.8 +/- 8.2 and 123.3 +/- 8.2, respectively) or CR (2.3 +/- 0.38 and 1.77 +/- 0.21, respectively). IV NAC reduced nephrotoxicity, (BUN 26.3 +/- 6.8, CR 0.47 +/- 0.15). NAC 50 mg/kg IA gave better protection than IV. In the repeated dose CDDP model, nephrotoxicity was blocked by 800 mg/kg NAC given IV but not IP. Blood concentrations of total NAC showed a dose response after IV NAC, but high dose NAC (1,200 mg/kg) by the PO route gave very low levels of NAC. Thus the protective properties of NAC are affected by the dose and route of administration. PMID- 17909807 TI - Phase I trial of PEG-interferon and recombinant IL-2 in patients with metastatic renal cell carcinoma. AB - PURPOSE: Pegylated interferon alpha-2b (PEG-Intron) is a conjugate of polyethylene glycol (PEG) and interferon alpha-2b, has a prolonged half-life, and an increased area under the curve (AUC) for interferon alpha-2b. The combination of PEG-Intron with recombinant interleukin-2 (rIL-2) was investigated in a phase 1 trial. To determine the maximal tolerable dose (MTD) and preliminary efficacy of concurrent subcutaneous (SC) administration of PEG-Intron and rIL-2 in patients with metastatic renal cell carcinoma (RCC). METHODS: Cohorts of 3-6 patients received escalating doses of PEG-Intron (I-1.5, II- 1.5, III-3.0, IV 3.0, V-4.5 microg/kg SC) given weekly in combination with rIL-2 administered three times weekly (TIW) for 6 weeks. rIL-2 dose levels were escalated in weeks 1 and 4 (I-10.0, II-15.0, III-15.0, IV-20.0, V-20.0 MIU/m(2) SC), and 5.0 MIU/m(2) SC TIW was administered during weeks 2, 3, 5 and 6. RESULTS: Thirty-four patients (24 men; 10 women) were accrued at dose levels I (n = 4), II (n = 4), III (n = 6), IV (n = 14), and V (n = 6) between October 2000 and October 2002. All but one patient had prior nephrectomy (n = 33) and all but one patient (97%) had received no prior systemic therapy. Patients received a median of four cycles of treatment (range 1-9). Dose limiting toxicity occurred at dose level V and included grade 4 neutropenia and hypoxemia. A partial response was found in 5 pts (15%). Median progression-free and overall survival were 9.0 (95% C.I. 5.6-13.1 months) and 31.9 months (95% C.I. 17.2-61.9 months), respectively. CONCLUSION: The combination of PEG-Interferon and SC rIL-2 can be administered with acceptable toxicity. PMID- 17909808 TI - The relative bioavailability of gefitinib administered by granular formulation. AB - BACKGROUND: Gefitinib (IRESSA) is normally administered as a once-daily oral tablet. However, many patients with head and neck cancer have difficulty swallowing medication in a tablet form. A granular formulation has recently been developed to facilitate the administration of gefitinib to patients who are unable to swallow tablets. OBJECTIVES: The aims of this study were to determine the relative bioavailability of a single dose of gefitinib when administered as 250 mg of a new granular formulation compared with the standard 250 mg tablet, and to assess the intra-subject variability of the granular formulation, in healthy subjects. METHODS: This was a randomized, open-label, three-period crossover study. Healthy male subjects (n = 18) received either a single gefitinib 250 mg tablet (once), or a 250 mg granular formulation of gefitinib (on two separate occasions) over the three dosing periods, in randomized order. Plasma concentrations of gefitinib were measured up to 240 h post-dose. RESULTS: The treatment ratio estimates for area under the plasma concentration versus time curve (AUC) and peak plasma concentration (C (max)) for the granular formulation when compared with the tablet were 1.05 (90% confidence intervals [CI] for the ratio 0.97-1.13) and 1.14 (90% CI for the ratio 1.01-1.28), respectively. The estimate for the intra-subject standard deviation for the granular formulation when given on 2 separate occasions was 0.143 for AUC and 0.165 for C (max), equivalent to a 1.4- and 1.7-fold intra-subject variability in AUC and C (max), compared with that observed for the tablet of two and threefold, respectively. CONCLUSIONS: There was little difference in exposure to gefitinib administered as the 250 mg granular formulation compared with the 250 mg standard tablet. The granular formulation of gefitinib could provide an alternative treatment regimen for patients unable or unwilling to swallow the standard tablet formulation, without compromizing exposure to gefitinib. PMID- 17909809 TI - The impact of delay in cryo-fixation on biomarkers of Src tyrosine kinase activity in human breast and bladder cancers. AB - Demonstration of pharmacodynamic activity of new, targeted cancer drugs in tumour tissue is potentially important in guiding early drug development. However, delays between tumour sampling and sample fixation may result in variability of pharmacodynamic biomarkers. The aim of this study, was to assess the impact of delays in fixation on biomarkers of Src kinase activity. A total of 20 patients with locally advanced breast cancer and 5 with early bladder cancer had multiple tissue samples taken which were fixed at documented time points up to 60 min after biopsy. These were examined to determine if the amount of Paxillin, phospho Paxillin, phospho-focal adhesion kinase (FAK) and total phospho-Tyrosine changed over time, using a quantitative lysate immunoassay. In breast cancer, there was an increase in the amount of phospho-Paxillin (60% per h; P = 0.019) up to 60 min after biopsy. The amount of total Paxillin decreased (28% per h; P = 0.034) over the same time course. In early bladder cancer, no changes were noted in any endpoints up to 45 min. Standardisation of the time taken between biopsy and fixation may be critical, particularly in studies using phosphorylated protein biomarkers. PMID- 17909810 TI - Phase II clinical study of gemcitabine in the treatment of patients with advanced nasopharyngeal carcinoma after the failure of platinum-based chemotherapy. AB - PURPOSE: This study was designed to evaluate the anti-tumor activity and toxicity profile of gemcitabine in the treatment of patients with advanced nasopharyngeal carcinoma (NPC) who had been pretreated with platinum-based chemotherapy. METHOD: This is an open label, single arm phase II trial. All patients were treated with single agent of gemcitabine. Gemcitabine was given in the dosage of 1.0 g/m(2) on days 1, 8, 15, each cycle repeated every 4 weeks. Gemcitabine was added to 100 ml normal saline infused over 30 min. RESULT: About 32 patients were enrolled in this trial. Thirty patients were assessable for response to treatment. Fourteen patients had a partial response (PR), giving an overall response rate of 43.8% (14/32); 9 patients had stable disease (28.1%) and 7 progressed disease (21.9%). The median time to progression was 5.1 months and median survival time was 16 months, 1 year survival rate was 67%, 2 year overall survival rate was 12%. A total of 11 patients (34.4%) experienced grade 3 and 4 toxicity and the main toxicity was myelosuppression. the non-hematology toxicity was minimal. CONCLUSION: The effectiveness of gemcitabine was higher and side effects were minimal in advanced NPC patients after platinum-based chemotherapy failed. PMID- 17909812 TI - Orally administered FTS (salirasib) inhibits human pancreatic tumor growth in nude mice. AB - BACKGROUND: S-trans,trans-farnesylthiosalicylic acid (salirasib, FTS) is a synthetic small molecule that acts as a potent Ras inhibitor. Salirasib inhibits specifically both oncogenically activated Ras and growth factor receptor-mediated Ras activation, resulting in the inhibition of Ras-dependent tumor growth. The objectives of this study were to develop a sensitive LC-MS/MS assay for determination of FTS in plasma, to assess the bioavailabilty of FTS after oral administration to mice, and then to examine the efficacy of orally administered FTS for inhibition of tumor growth in a nude mouse model. METHODS: FTS was isolated from mouse plasma by liquid chromatography on a Columbus 5-mum particle size, 50 x 2 mm id column with a methanol/5 mM ammonium acetate (80/20) mobile phase (isocratic elution) at a flow rate of 0.3 ml/min. MS/MS was performed on a PE Sciex API 365 with Turbo Ion Spray as interface and negative ion ionization; parent ion (m/z): 357.2; daughter ion (m/z) 153.2; retention time 2.3 min. For plasma analysis, the amount of analyte in each sample was calculated by comparing response of the analyte in that sample to a nine-point standard curve linear over the range 3-1000 ng/ml. Pharmacokinetic studies were performed in mice following intraperitoneal dosing (20 mk/kg in PBS) or oral dosing (40 mg/kg in either 0.5% aqueous CMC or corn oil). Panc-1 tumor growth in nude mice was determined following daily oral dosing with FTS in 0.5% CMC (40, 60, or 80 mg/kg), or in combination with weekly gemcitabine (30 mg/kg). RESULTS: Salirasib was readily detected in mouse plasma by LC-MS/MS at a detection limit of 3 ng/ml. For each route of administration, t (max) was 1 h and t (1/2) ranged from 1.86 to 2.66 h. Compared to IP administration, the oral bioavailabilty of FTS was 69.5% for oral CMC and 55% for oral corn oil suspensions, while clearance and volume of distribution were higher in both oral preparations. The orally administered salirasib inhibited panc-1 tumor growth in a dose dependent manner (67% reduction in tumor weight at the highest dose, P < 0.002 vs. control, n = 10 mice per group) and at a 40 mg/kg daily dose was synergistic with gemcitabine (83% increase in survival rate, n = 8 mice per group). CONCLUSIONS: Salirasib exhibits good bioavailabilty after oral administration, as determined by a highly sensitive method for quantification in plasma. The orally available Ras inhibitor salirasib inhibited growth in nude mice, and may thus be considered for clinical trials. PMID- 17909811 TI - A limited sample model to predict area under the drug concentration curve for 17 (allylamino)-17-demethoxygeldanamycin and its active metabolite 17-(amino)-17 demethoxygeldanomycin. AB - PURPOSE: The Hsp90-directed anticancer agent 17-(allylamino)-17 demethoxygeldanamycin (17-AAG) is currently undergoing phase I and phase II clinical investigation. Our goal was to develop a simple limited sampling model (LSM) for AUC of 17-AAG and its active metabolite, 17-(amino)-17 demethoxygeldanomycin (17-AG) using drug concentrations from a few time points. METHODS: Pharmacokinetic data from 34 patients treated at 11 dose levels on a Mayo Clinic Cancer Center phase I clinical trial of 17-AAG was utilized. Blood samples were collected at 11 different time points, spanning 25 h. Graphical methods and correlations were used to assess functional forms and univariate relationships. Multivariate linear regression and bootstrap resampling were used to develop the LSM. RESULTS: Using log-transformed data, the two and three time point 17-AAG LSMs are log-AUC (17-AAG) = 0.869 + 0.653*(C(55min)) +0.469*(C(5h)) and log-AUC (17-AAG) = 2.449 + 0.400*(C(55min)) +0.441*(C(5h)) +0.142*(C(9h)). The two and three time point LSMs for 17-AG are log-AUC (17-AG) = 3.590 + 0.747*(C(5h)) +0.169*(C(17h)), and log-AUC (17-AG) = 3.797 + 0.650*(C(5h)) +0.111*(C(9h)) +0.122*(C(17h)). Ninety-seven percent and 94% of the predicted log AUC values were within 5% of the observed log-AUC for the two and three time point models for 17-AAG and 17-AG respectively. CONCLUSIONS: The precise calculation of AUC is cumbersome and expensive in terms of patient and clinical resources. The LSM developed using a multivariate regression approach is clinically and statistically meaningful. Prospective validation is underway. PMID- 17909814 TI - Stochastic models for phylogenetic trees on higher-order taxa. AB - Simple stochastic models for phylogenetic trees on species have been well studied. But much paleontology data concerns time series or trees on higher-order taxa, and any broad picture of relationships between extant groups requires use of higher-order taxa. A coherent model for trees on (say) genera should involve both a species-level model and a model for the classification scheme by which species are assigned to genera. We present a general framework for such models, and describe three alternate classification schemes. Combining with the species level model of Aldous and Popovic (Adv Appl Probab 37:1094-1115, 2005), one gets models for higher-order trees, and we initiate analytic study of such models. In particular we derive formulas for the lifetime of genera, for the distribution of number of species per genus, and for the offspring structure of the tree on genera. PMID- 17909813 TI - Boltzmann ensemble features of RNA secondary structures: a comparative analysis of biological RNA sequences and random shuffles. AB - Ensemble-based approaches to RNA secondary structure prediction have become increasingly appreciated in recent years. Here, we utilize sampling and clustering of the Boltzmann ensemble of RNA secondary structures to investigate whether biological sequences exhibit ensemble features that are distinct from their random shuffles. Representative messenger RNAs (mRNAs), structural RNAs, and precursor microRNAs (miRNAs) are analyzed for nine ensemble features. These include structure clustering features, the energy gap between the minimum free energy (MFE) and the ensemble, the numbers of high-frequency base pairs in the ensemble and in clusters, the average base-pair distance between the MFE structure and the ensemble, and between-cluster and within-cluster sums of squares. For each of the features, we observe a lack of significant distinction between mRNAs and their random shuffles. For five features, significant differences are found between structural RNAs and random counterparts. For seven features including the five for structural RNAs, much greater differences are observed between precursor miRNAs and random shuffles. These findings reveal differences in the Boltzmann structure ensemble among different types of functional RNAs. In addition, for two ensemble features, we observe distinctive, non-overlapping distributions for precursor miRNAs and random shuffles. A distributional separation can be particularly useful for the prediction of miRNA genes. PMID- 17909815 TI - The spectrum of spontaneous mutations caused by deficiency in proteasome maturase Ump1 in Saccharomyces cerevisiae. AB - Ump1 is responsible for maturation of the catalytic core of the 26S proteasome. Dysfunction of Ump1 causes an increase in the frequency of spontaneous mutations in Saccharomyces cerevisiae. In this study we analyze the spectrum of mutations occurring spontaneously in yeast deficient in Ump1 by use of the SUP4-o system. Single base substitutions predominate among the mutations analyzed (73 of the 91 alterations examined). Two major classes are GC to TA transversions and GC to AT transitions ( approximately 50 and approximately 30% of base substitutions, respectively). Besides base substitutions, almost all the major types of sequence alterations are represented. The specificity and distribution of mutations occurring in the ump1 strain are unique compared to the spectra previously established for other yeast mutators. However, the profile of mutations arising in this strain is similar to that observed in wild type. The same similarity has previously been reported for yeast deficient in Mms2, a protein involved in Rad6 dependent postreplication DNA repair (PRR). The specificity of the mutator effect caused by ump1 is discussed in light of the proposed role of the proteasome activity in the regulation of the PRR mechanisms. PMID- 17909816 TI - Radiation dose estimates in dual-source computed tomography coronary angiography. AB - The purpose of this study was to quantify radiation dose parameters of dual source CT coronary angiography. Eighty patients underwent contrast-enhanced, retrospectively ECG-gated dual-source CT coronary angiography with heart rate adapted ECG pulsing using two algorithms: In 40 patients, the tube current was reduced to 20% (A(min1)) of the normal tube current (A(max)) outside the pulsing window; in 40 patients tube current was reduced to 4% (A(min2)) of A(max). Mean CTDI(vol) in the A(min1) group was 45.1 +/- 3.6 mGy; the mean CTDI(vol) in the A(min2) group was 39.1 +/- 3.2 mGy, with CTDI(vol) in the A(min2) group being significantly reduced when compared to the A(min1) group (P < 0.001). A significant negative correlation was found between CTDI(vol) and heart rate in group A(min1) (r = -0.82, P < 0.001), whereas no correlation was found between CTDI(vol) and heart rate in group A(min2) (r = -0.066). Using the conversion coefficient for the chest, dual-source CT coronary angiography resulted in an estimated mean effective dose of 8.8 mSv in the A(min1) group and 7.8 mSv in the A(min2). Radiation exposure of dual-source CT coronary angiography using an ECG pulsing protocol reducing the tube current to 20% significantly decreases with increasing heart rates, despite using wider pulsing windows at higher heart rates. When using a protocol with reduced tube current of 4%, the radiation dose is significantly lower, irrespective of the heart rate. PMID- 17909817 TI - Quantitative assessment of left ventricular function with dual-source CT in comparison to cardiac magnetic resonance imaging: initial findings. AB - Cardiac magnetic resonance imaging and echocardiography are currently regarded as standard modalities for the quantification of left ventricular volumes and ejection fraction. With the recent introduction of dual-source computedtomography (DSCT), the increased temporal resolution of 83 ms should also improve the assessment of cardiac function in CT. The aim of this study was to evaluate the accuracy of DSCT in the assessment of left ventricular functional parameters with cardiac magnetic resonance imaging (MRI) as standard of reference. Fifteen patients (two female, 13 male; mean age 50.8 +/- 19.2 years) underwent CT and MRI examinations on a DSCT (Somatom Definition; Siemens Medical Solutions, Forchheim, Germany) and a 3.0-Tesla MR scanner (Magnetom Trio; Siemens Medical Solutions), respectively. Multiphase axial CT images were analysed with a semiautomatic region growing algorithms (Syngo Circulation; Siemens Medical Solutions) by two independent blinded observers. In MRI, dynamic cine loops of short axis slices were evaluated with semiautomatic contour detection software (ARGUS; Siemens Medical Solutions) independently by two readers. End-systolic volume (ESV), end diastolic volume (EDV), ejection fraction (EF) and stroke volume (SV) were determined for both modalities, and correlation coefficient, systematic error, limits of agreement and inter-observer variability were assessed. In DSCT, EDV and ESV were 135.8 +/- 41.9 ml and 54.9 +/- 29.6 ml, respectively, compared with 132.1 +/- 40.8 ml EDV and 57.6 +/- 27.3 ml ESV in MRI. Thus, EDV was overestimated by 3.7 ml (limits of agreement -46.1/+53.6), while ESV was underestimated by 2.6 ml (-36.6/+31.4). Mean EF was 61.6 +/- 12.4% in DSCT and 57.9 +/- 9.0% in MRI, resulting in an overestimation of EF by 3.8% with limits of agreement at -14.7 and +22.2%. Rank correlation rho values were 0.81 for EDV (P = 0.0024), 0.79 for ESV (P = 0.0031) and 0.64 for EF (P = 0.0168). The kappa value of inter-observer variability were amounted to 0.85 for EDV, ESV and EF. DSCT offers the possibility to quantify left ventricular function from coronary CT angiography datasets with sufficient diagnostic accuracy, adding to the value of the modality in a comprehensive cardiac assessment. The observed differences in the measured values may be due to different post-processing methods and physiological reactions to contrast material injection without beta-blocker medication. PMID- 17909818 TI - Stereotactic large-core needle breast biopsy: analysis of pain and discomfort related to the biopsy procedure. AB - The purpose of this study was to determine the significance of variables such as duration of the procedure, type of breast tissue, number of passes, depth of the biopsies, underlying pathology, the operator performing the procedure, and their effect on women's perception of pain and discomfort during stereotactic large core needle breast biopsy. One hundred and fifty consecutive patients with a non palpable suspicious mammographic lesions were included. Between three and nine 14 gauge breast passes were taken using a prone stereotactic table. Following the biopsy procedure, patients were asked to complete a questionnaire. There was no discomfort in lying on the prone table. There is no relation between type of breast lesion and pain, underlying pathology and pain and performing operator and pain. The type of breast tissue is correlated with pain experienced from biopsy (P = 0.0001). We found out that patients with dense breast tissue complain of more pain from biopsy than patients with more involution of breast tissue. The depth of the biopsy correlates with pain from biopsy (P = 0.0028). Deep lesions are more painful than superficial ones. There is a correlation between the number of passes and pain in the neck (P = 0.0188) and shoulder (P = 0.0366). The duration of the procedure is correlated with pain experienced in the neck (P = 0.0116) but not with pain experienced from biopsy. PMID- 17909819 TI - Vascular clamp stabilization of pylorus during laparoscopic pyloromyotomy. AB - We describe a technique of grasping the pylorus during laparoscopic pyloromyotomy using a percutaneously inserted vascular clamp. The use of the vascular clamp results in better visualization and stabilization during laparoscopic pyloromyotomy. PMID- 17909820 TI - Capecitabine vs continuous infusion 5-FU in neoadjuvant treatment of rectal cancer. A retrospective review. AB - BACKGROUND: Standard therapy for locally advanced rectal cancer (LARC) is concurrent neo-adjuvant chemo-radiation using infusional 5-fluorouracil (CIV-5 FU). Capecitabine (CAP) offers a convenient oral replacement for CIV-5-FU. There is no randomized trial comparing infusional 5-FU to capecitabine. We retrospectively compared the safety and efficacy of CAP-based regimens with well established CIV-5-FU-based regimens in LARC. MATERIALS AND METHODS: We collected published data on 542 patients treated on either CIV-5-FU (197) or CAP (345) with concurrent radiation (external radiation treatment, XRT) for LARC. This included Phase I or II studies published or available from Pubmed. Safety was assessed by determining proportion of patients who experienced grade III/IV adverse effects. Efficacy was assessed by determining pathological complete response (pCR). Chi square tests were used to compare the two regimens. A P value less than 0.05 was considered statistically significant. Statistical tests were further corrected for multiplicity using the method of Benjamini and Yekutieli (Ann Stat, 29(4):1165-1188, 2001). RESULTS: pCR was significantly higher in patients getting CAP vs CIV-5-FU (25 vs 13%; P = 0.008,.P adj = 0.034). Both regimens were generally well tolerated. There was no grade IV toxicity reported. Grade III hand foot syndrome was more common in the CAP group, and grade III diarrhea was more common in the CIV group. CONCLUSIONS: CAP when compared to CIV seems to have superior efficacy with reasonable toxicities. It is reasonable to treat LARC with CAP + XRT. PMID- 17909822 TI - Surface coating to improve the metal-cement bonding in cemented femur stems. AB - INTRODUCTION: Hydrolytic debonding of the metal-cement interface is one of the main reasons for aseptic loosening in cemented hip arthroplasty. MATERIALS AND METHODS: BiContact femur stems (CoCrMo-/TiAl6V4-alloy) were coated by a silica/silane interlayer coating system. The stems were cemented into artificial femurs. The cyclical loading (DIN ISO 7206-4) was performed within a hip simulator. Uncoated stems (CoCrMo-/TiAl6V4-alloy) were prepared and loaded the same way. After loading, the metal-cement and the bone-cement interfaces were analysed. Unloaded uncoated and unloaded coated BiContact stems served as a control. RESULTS: The coated loaded stems showed a significant reduction in debonding and cement failure (P < or = 0.05). A high correlation was documented between debonding and cement failure (rSpear> or = 0.9). There was no significant difference between CoCrMo- and TiAl6V4-stems (P > or = 0.05). CONCLUSION: The silica/silane coating significantly decreased hydrolytic debonding at the metal bone cement interface with consecutively less cement failure. PMID- 17909823 TI - Revision total hip arthroplasty with an uncemented primary stem in 79 patients. AB - INTRODUCTION: Revision in THA continues to be a technical challenge because of difficulties in fixation of the femoral component in mostly deficient bone in the proximal femur. In cases with minor cortical defects, the use of primary stems in revision surgery has also been described by some authors. MATERIALS AND METHODS: Seventy-nine patients with minor femoral bone defects were reviewed retrospectively (mean follow-up 6.8 +/- 3.9 years), who underwent a femoral component revision surgery using the uncemented primary Bicontact stem (Aesculap, Tuttlingen, Germany). Furthermore, the radiographs (anteroposterior and lateral) before, after surgery and at latest follow-up were analysed concerning femoral defects, proximal bone loss, and to determine the quality of bony fixation. RESULTS: The average Harris hip score (HHS) was 42.2 +/- 20.8 preoperative and improved to 78.9 +/- 12.5 at latest follow-up (p < 0.001). Motion Score increased significantly from 2.7 +/- 1.9 to 3.5 +/- 1.4 (p < 0.05) and pain score decreased significantly from 5.7 +/- 2.9 to 3.6 +/- 2.4 (p = 0.005). During follow-up there were only four re-revisions within 2 years after revision. The results and clinical outcome of this study correspond to those published before, using primary cementless stems in cases of revision. CONCLUSION: Therefore, the primary uncemented Bicontact stem appears to be a good alternative to other revision systems in well-selected femoral revision cases with minor defects. PMID- 17909824 TI - Relationship of the menstrual cycle phase to anterior cruciate ligament injuries in teenaged female athletes. AB - INTRODUCTION: Anterior cruciate ligament (ACL) injuries are more common among female athletes compared to male athletes. Several studies have been reported to explain the gender difference in ACL injury rates and several risk factors underlying gender disparity are believed to exist. Hormonal effects are considered to be one of the etiological factors for female non-contact ACL injuries. The objectives of this study were to determine if ACL injuries occurred randomly or correlated with a specific phase of the female menstrual cycle in teenaged female athletes and then to determine if pre-menstrual and menstrual dysfunctions influenced these ACL injuries. MATERIALS AND METHODS: Eighteen of 37 consecutive female athletes, with ACL injuries, met the study criteria: teenage, with regular menstrual cycle, and non-contact injury. The menstrual history, athletic activity, and injury history were collected. RESULTS: Seventy-two percent of the subjects had premenstrual symptoms and 83% had menstrual symptoms. The subjective activity level at the follicular phase was significantly lowest between the phases. A significant statistical association was found between the phase of the menstrual cycle and ACL injuries (P = 0.0002). There were more injuries in the ovulatory phase than expected, and fewer injuries occurred in the other phases. CONCLUSION: The results showed a significant increase in non contact ACL injuries in teenage female athletes during the ovulatory phase of the menstrual cycle and the subjective activity level and the premenstrual and menstrual symptoms might not affect the likelihood of the injuries. These findings suggest that sex hormones might play a role in the incidence of female non-contact ACL injuries. PMID- 17909825 TI - Clinical outcome of closed isolated subtalar dislocations. AB - INTRODUCTION: Subtalar dislocation (SD) is an uncommon injury accounting for 1-2% of all dislocations. It involves simultaneous disruption of the talocalcaneal and talonavicular joints, without involvement of the calcaneocuboid or tibiotalar joints or talar neck fracture. We present a retrospective study of pure medial and lateral SDs treated conservatively and discuss the pathogenesis, classification, prognostics and therapeutic aspects of SD. MATERIALS AND METHODS: Thirty patients, 24 men and 6 women (mean age 33 years; range 18-55) with closed isolated SD were treated conservatively and re-evaluated at 5-12 years. There were 20 medial and 10 lateral dislocations. All patients were managed with immediate closed reduction under general anaesthesia. Open dislocations and SDs associated with fractures were excluded. RESULTS: The mean AOFAS Ankle-Hindfoot score was 78.8. Seven patients (all with medial SDs) had an AOFAS score of 100; 14 patients (11 with medial and 3 with lateral SD) had a mean AOFAS score of 85; 6 patients (three with medial and three with lateral SD) had a mean AOFAS score of 65; and 3 patients (all with lateral SDs) had a mean AOFAS score of 28. The latter patients subsequently underwent subtalar fusion, with a fair outcome. The mean AOFAS scores of patients with lateral and medial SD were not significantly different (P = 0.05). CONCLUSION: Various factors adversely affect outcome, including type of dislocation (lateral/medial, open/closed), severity of the injury, associated fractures, length of immobilization. Management of closed isolated SD is by immediate conservative treatment in order to avoid or reduce the incidence of early soft-tissue and vascular complications and poor long-term outcomes due to post-traumatic arthritis, talus necrosis and subtalar joint stiffness. However, complications may still arise despite correct treatment. PMID- 17909826 TI - Microvascular decompression of cochlear nerve for tinnitus incapacity: pre surgical data, surgical analyses and long-term follow-up of 15 patients. AB - The level of success of neurovascular decompression in ponto-cerebellar angle for hemifacial spasm and trigeminal neuralgia has already established the reality of the pathology to explain such symptoms. However, cochlear nerve compression syndrome by vascular loop is still a controversial topic. We have performed a retrospective cases review with long-term follow-up (5-7 years) concerning the results of microvascular decompression surgery of the cochlear nerve via an endoscopy assisted retrosigmoid approach on 15 patients suffering from unilateral incapacitating tinnitus with abnormal auditory brainstem response and an offending vessel on magnetic resonance imaging. During the surgery, a vascular compression was found on every patient. In a long-term follow-up, 53.3% (8 cases) of our tinnitus cases improved and 20% (3 cases) of them were completely cured. The ABR returned to normal in all patients who had good clinical results (diminished or disappeared tinnitus). When a vertebral artery loop (5 cases) was concerned we obtained 80% of good clinical results. No one showed amelioration or sudden aggravation of their hearing. Three cases required surgical correction of cerebrospinal fluid leak and one case developed spontaneously regressive swallowing problems. Such microvascular decompression surgery of the cochlear nerve appears to be successful in treating incapaciting tinnitus in particular when a vertebral artery loop is observed. Therefore, in such a case, one might recommend neurovascular decompression surgery, keeping in mind that the complications of this surgery should be minimized by a careful closure of the retrosigmoid approach. In order to ensure a better selection of patient more accurate cochlear nerve monitoring and functional MRI should be a promising assessment. PMID- 17909827 TI - Extra- and intracranial dumbbell-shaped hemangiopericytoma. AB - Hemangiopericytomas are malignant tumors arising from pericytic cells and account for less than 1% of all vascular neoplasms. We report a rare case of an extra- and intracranial dumbbell-shaped hemangiopericytoma originating from the soft tissue of the neck and penetrating the skull base with invasion into the posterior cranial fossa. The 59-year-old female patient presented with a large pulsating neck mass and reported weakness, abnormal fatigue and headache. MRI revealed an inhomogeneously enhancing tumor and cerebral angiography showed intensive vascularization. Preoperative embolization was performed in order to decrease the operative blood loss. The tumor was operated via a far lateral approach through an osteoclastic suboccipital craniotomy. Total resection of both the intra- and extracranial part of the neoplasm (grade I by Simpson) could be achieved. The histopathological analysis revealed a mesenchymal, hypervascular tumor with the classic staghorn vascular pattern. In this article, we discuss the clinical presentation and multidisciplinary management of hemangiopericytoma and describe the radiological and pathological features of this tumor entity. PMID- 17909828 TI - Sentinel lymph node biopsy or elective neck dissection for patients with oral squamous cell carcinoma? AB - Sentinel lymph node biopsy (SNB) seems to be a promising method for staging clinically N0 neck in patients with oral squamous cell carcinoma (OSCC). In the present study, SNB was performed on 46 patients having elective neck dissection (END; six bilateral dissections) for T1-T3N0 OSCC. Sentinel lymph nodes (SLN) were first examined according to only slightly modified standard histopathologic protocol including sections at 1-2 mm intervals and H&E staining. SLN that appeared false negative (i.e. metastatic non-SLN without metastasis in a SLN) after the initial histopathologic examination were further assessed by step sectioning at 150 microm intervals and immunohistochemistry. Of the 47 neck sides with at least one SLN identified, nine contained metastasis in nine patients. After the initial histopathologic examination, SLNs were negative for malignant cells in four out of the nine metastatic neck sides. In one neck side, two metastatic SLNs were detected after the additional meticulous histopathologic work-up of the initially false negative SLNs. Therefore, in three neck sides the SLN did not contain metastasis although there was a metastasis in a non-SLN. In all these three cases with a false negative SLN, only one SLN had been identified. The sensitivity of the method (employing extensive histopathologic work-up) for detection of occult cervical metastasis was 67% (6/9 neck sides). The sensitivity of SNB for detection of occult metastasis seems to be poor in cases where only one SLN can be identified. The results of this study do not entitle us to entirely replace END by SNB in patients with OSCC. PMID- 17909829 TI - Microdebrider-assisted partial tonsillectomy: short- and long-term outcomes. AB - Microdebrider-assisted partial tonsillectomy has gained popularity in recent years. However, no prospective long-term follow-up exists in the literature and the risk of increase in tonsillitis is still a concern. We conducted a prospective randomized controlled study to assess the short-term benefits of microdebrider-assisted partial tonsillectomy (group 1) compared to electrocautery assisted total tonsillectomy (group 2), to monitor the durability of the improvement and watch for any change in the rate of tonsillitis. Patients with symptomatic tonsillar hyperplasia were included and underwent either technique of tonsillar surgery. Duration of operation, amount of intra-operative blood loss, immediate and late complications, postoperative pain, return to normal activity and diet, and relief of obstructive symptoms were measured. Recurrence of symptoms and change in rate of tonsillitis in group 1 were monitored. We studied 143 patients, 77 in group 1 and 66 in group 2. There was no significant difference in the surgical time (P>0.05) or postoperative bleeding (P>0.05) but more blood loss in group 1 (P<0.05) and more dehydration in group 2 (P<0.05) were encountered. All patients had complete relief of symptoms. Group 1 returned earlier to normal activity (2.19 vs. 5.71 days; P<0.05), to normal diet (5.28 vs. 8.16 days; P<0.05) and needed less frequent analgesics (2.14 vs. 6.1 days; P<0.05). More than two-thirds of group 1 and less than one-third of group 2 were pain free after day 3. Most group 1 parents (96.1%) were highly satisfied, at initial follow-up, regarding the decision to perform the surgery in contrast to group 2 parents (19.7%). No recurrence of symptoms and no increase in rate of tonsillitis were noticed among group 1 after 20 months mean follow-up (median 20.6, range 1-36.2 months). Group 1 showed short-term benefits over group 2 and maintained the resultant improvement on the long-term with no infectious drawbacks. PMID- 17909830 TI - PMMA (polymethylmetacrylate) microspheres and stabilized hyaluronic acid as an injection laryngoplasty material for the treatment of glottal insufficiency: in vivo canine study. AB - Both PMMA (polymethylmetacrylate) microspheres (PM) and stabilized hyaluronic acid (HA) are recently used for facial augmentation. The aim of this study was to test functional effect, durability, and safety of the injection of these two materials into true vocal folds, and test their availability as injection laryngoplasty materials in vivo canine model. The study was carried out with 16 beagle dogs (8 males and 8 females, average weight of 12.4 kg). No biological difference was detected between two groups; PM (Artecoll) injection and HA (Restylane) injection group. After inducing complete unilateral paralysis of the recurrent laryngeal nerve of the dogs, either PM or HA was injected into the paralyzed vocal fold. One, 3, 6, and 9 months after the injection, clinical outcomes and videostroboscopic findings were evaluated by investigators who were blind to the injection materials. Histological study and microscopic computerized augmentation dimension analysis were also performed. In HA injection group, up to 30% the HA was gradually resorbed over time. However, in PM group, the dimension of the augmented region after 9 months was similar to that after 1 month. In both groups, the mucosal waves of the vocal folds decreased in amplitude and periodicity, but they were still well detected during the follow-up periods. Acute immune reaction to HA was not detected, but some degree of foreign body reaction occurred in PM injection group. Both PM and HA are safe and relatively durable in vocal folds and they are considered as useful candidates for injection laryngoplasty. PMID- 17909831 TI - Glottic versus supraglottic tumors: differential molecular profile. AB - Glottis and supraglottis, although anatomically interconnected, are embryologically distinct. Moreover, squamous cell carcinomas arising from these subsites, differ in terms of epidemiology, risk factors, clinical behaviour and prognosis. This study aims to explore any possible differences between their molecular profiles. We investigated in the two tumor types, the expression of epidermal growth factor receptor (EGFR), nuclear factor-kappaB (NF-kappaB) and retinoid X receptor alpha (RXRalpha), principal signal transducers associated with cancer, as well as cyclooxygenase-2 (COX-2), an enzyme induced in malignant neoplasms. The clinical material includes tumor specimens from 61 patients with laryngeal cancer of glottic or supraglottic origin. Subsite groups were matched for gender, age and histological grade. Paraffin-section immunohistochemistry was performed, to detect the aforementioned molecules. Staining patterns were membranic and cytoplasmic for EGFR, purely cytoplasmic for COX-2, nuclear for RXRalpha and cytoplasmic, as well as nuclear, for NF-kappaB. Intense EGFR and RXRalpha expression was significantly associated with glottic tumor descent (P = 0.011 and 0.001, respectively). No significant relationship was established between neoplasm location and expressions of NF-kappaB, COX-2. Our results show that tumors emerging from the two laryngeal regions, are different with regard to their molecular constitution. Upregulation of EGFR and RXRalpha in carcinomas of the glottis, might be important in the design of subsite-specific chemotherapeutic approaches. PMID- 17909832 TI - Functional links between Drosophila Nipped-B and cohesin in somatic and meiotic cells. AB - Drosophila Nipped-B is an essential protein that has multiple functions. It facilitates expression of homeobox genes and is also required for sister chromatid cohesion. Nipped-B is conserved from yeast to man, and its orthologs also play roles in deoxyribonucleic acid repair and meiosis. Mutation of the human ortholog, Nipped-B-Like (NIPBL), causes Cornelia de Lange syndrome (CdLS), associated with multiple developmental defects. The Nipped-B protein family is required for the cohesin complex that mediates sister chromatid cohesion to bind to chromosomes. A key question, therefore, is whether the Nipped-B family regulates gene expression, meiosis, and development by controlling cohesin. To gain insights into Nipped-B's functions, we compared the effects of several Nipped-B mutations on gene expression, sister chromatid cohesion, and meiosis. We also examined association of Nipped-B and cohesin with somatic and meiotic chromosomes by immunostaining. Missense Nipped-B alleles affecting the same HEAT repeat motifs as CdLS-causing NIPBL mutations have intermediate effects on both gene expression and mitotic chromatid cohesion, linking these two functions and the role of NIPBL in human development. Nipped-B colocalizes extensively with cohesin on chromosomes in both somatic and meiotic cells and is present in soluble complexes with cohesin subunits in nuclear extracts. In meiosis, Nipped-B also colocalizes with the synaptonemal complex and contributes to maintenance of meiotic chromosome cores. These results support the idea that direct regulation of cohesin function underlies the diverse functions of Nipped-B and its orthologs. PMID- 17909833 TI - Analysis of Y-chromosomal SNP haplogroups and STR haplotypes in an Algerian population sample. AB - The distribution of Y-chromosomal single nucleotide polymorphism (SNP) haplogroups and short tandem repeat (STR) haplotypes was determined in a sample of 102 unrelated men of Arab origin from northwestern Algeria (Oran area). A total of nine different haplogroups were identified by a panel of 22 binary markers. The most common haplogroups observed in the Algerian population were E3b2 (45.1%) and J1 (22.5%). Y-STR typing by a 17-loci multiplex system allowed 93 haplotypes to be defined (88 were unique). Striking differences in the allele distribution and gene diversity of Y-STR markers between haplogroups could be found. In particular, intermediate alleles at locus DYS458 specifically characterized the haplotypes of individuals carrying haplogroup J1. All the intermediate alleles shared a common repeat sequence structure, supporting the hypothesis that the variant originated from a single mutational event. PMID- 17909834 TI - Variables affecting the probability of complete fusion of the medial clavicular epiphysis. AB - In this study, we have combined data on clavicle fusion from different studies and applied a binomial logistic regression analysis. As such, we aimed to assess whether or not variables such as sex, socioeconomic status, and ethnicity influence the probability of having mature, i.e., completely fused clavicles at a given age. We further explored whether the method of clavicle examination, i.e., diagnosis from either a dry bone specimen, an examination of X-rays, or an examination of computed tomography scans, affects the probability of being diagnosed with mature clavicles. It appeared that only ethnicity did not significantly affect this probability. Finally, we illustrated how the logit model may be used to predict the probability of being diagnosed with mature clavicles. PMID- 17909835 TI - Blockade of VEGFR3-signalling specifically inhibits lymphangiogenesis in inflammatory corneal neovascularisation. AB - PURPOSE: Inflammatory corneal hem- and lymphangiogenesis occurring both prior to as well as after penetrating keratoplasty significantly increase the risk for subsequent immune rejections. The purpose of this study was to analyze whether the blocking anti-VEGFR3 antibody mF4-31C1 is able to inhibit the outgrowth of pathologic new lymphatic vessels in a mouse model of suture-induced, inflammatory corneal neovascularisation, and whether this antibody specifically inhibits lymphangiogenesis without affecting hemangiogenesis. METHODS: Three interrupted 11-0 nylon sutures were placed into the corneal stroma of BALB/c mice (6 weeks old) and left in place for 7 days to induce neovascularisation. The treatment group (n = 9) received the anti-VEGFR3 antibody mF4-31C1 intraperitoneally on the day of surgery and 3 days later (0.5 mg/mouse). Control mice received an equal amount of control IgG solution. For immunohistochemistry, corneal flat mounts were stained with LYVE-1 as a specific lymphatic vascular endothelial marker and with CD31 as panendothelial marker. Morphometry was performed with the image analysis software analySIS;B (Soft Imaging Systems GmbH, Munster, Germany). To improve the objectivity and precision of the morphometrical analysis, we established a modified method using grey filter sampling on monochromatic pictures. RESULTS: The mF4-31C1 antibody-treated mice displayed nearly complete inhibition of lymphangiogenesis compared with IgG controls (p < 0.006). In contrast, there was no significant inhibitory effect observed with respect to blood vessel growth (p > 0.05). CONCLUSIONS: Inflammatory corneal lymphangiogenesis seems to depend on VEGFR3-signalling. By blocking this receptor the ingrowths of lymphatic vessels can be inhibited almost completely, and specifically without affecting hemangiogenesis. This may open new treatment options to promote (corneal) graft survival without affecting hemangiogenesis. PMID- 17909837 TI - Syndromes from segmental vibration and nerve entrapment: observations on case definitions for carpal tunnel syndrome. AB - OBJECTIVES: The purpose of this paper is to assess the overlap and stability of two different case definitions of carpal tunnel syndrome CTS. The analysis considers the association between different case definitions and objective tests (sensory nerve conduction velocities, SNCVs and vibrotactile perception thresholds, TTS), and the natural history of CTS, in the context of two vibration exposed cohorts. METHODS: Clinical CTS cases were defined in two ways: (1) by the study physician using fixed criteria, and; (2) by questionnaire and hand diagram. SNCV in median and ulnar nerves was measured for digital, transpalmar, and transcarpal segments, and conventionally as from wrist-digit. Skin temperature was assessed as a point measurement by thermistor and regionally by thermal imaging. VTTs were determined at the bilateral fingertips of the third and fifth digits using a tactometer meeting the requirements of ISO 13091-1 (ISO 2001). The subjects were cohorts of shipyard workers in 2001 and 2004, and dental hygienists in 2002 and 2004. RESULTS: Results are reported for 214 shipyard workers in 2001 and 135 in 2004, and for 94 dental hygienists in 2002 and 66 in 2004. In 2001, 50% of shipyard workers were diagnosed as CTS cases by at least one of the diagnostic schemes, but only 20% were positive by both criteria. Among study physician diagnosed cases, 64% were CTS negative in 2001, 76% were negative in 2004, 13% were positive in both years, 22% became negative after being positive, and 11% became positive after being negative. For only study physician diagnosed CTS did VTTs differ between cases differ and non-cases in digit 3; there was no such distinction in digit 5. The dental hygienists had little CTS. CONCLUSION: Clinical case definitions of CTS based on diagrams and self-assessment, and clinical evaluation have limited overlap. Combining clinical criteria to create a more narrow or specific case definition of CTS does not appear to predict SNCV. The natural history of CTS suggests a protean disorder with considerable flux in case status over time. PMID- 17909838 TI - Coronary risks among seafarers aboard German-flagged ships. AB - OBJECTIVES: Cardiovascular diseases belong to the major causes of maritime service disablement (approximately 18%). The aim of this study was to investigate the frequency of coronary risk factors in seamen on vessels sailing under the German flag and to assess the risk of coronary heart disease (CHD) dependent on job-related factors. METHODS: Within a medical surveillance program, a cross sectional study with interview, blood sampling, and blood pressure measurements was conducted among a total of 205 male seafarers sailing under the German flag (response 84.9%). Due to missing blood analysis, 13 seamen were excluded. The predicted 10-year risk of an acute coronary event of a study subgroup of 45 German seafarers was compared to the corresponding risk of a sample of the German working population (PROCAM study). RESULTS: In the total sample (n = 161), 55 seamen (34.2%) had at least three CHD risk factors. The most prominent independent CHD risk factors in seafarers were hypertension (49.7%), high triglycerides (41.6%), older age (39.8%), and smoking (37.3%). Compared with non Europeans, European seafarers were about twice as likely to have more than three risk factors after adjusting for age [OR 2.4 (95% CI 1.01-4.55)]. Particularly engine room officers and galley/operating staff were at a higher coronary risk. After standardizing for age, the German seamen investigated showed a similar predicted 10-year CHD risk as the German population of about the same age working ashore of the PROCAM study. CONCLUSIONS: The CHD risks in seafarers should be reduced by low-fat diets, anti-smoking campaigns and blood pressure control/treatment. In spite of the seafarers' regular medical surveillance examination, their CHD risk was comparable to a reference population working ashore. Our results support the hypothesis that working on vessels may augment the risk of CHD. PMID- 17909839 TI - Diagnosing soft tissue rheumatic disorders of the upper limb in epidemiological studies of vibration-exposed populations. AB - OBJECTIVES: To investigate approaches adopted to diagnose soft tissue rheumatic disorders of the upper limb (ULDs) in vibration-exposed populations and in other settings, and to compare their methodological qualities. METHODS: Systematic searches were made of the Medline, Embase, and CINAHL electronic bibliographic databases, and of various supplementary sources (textbooks, reviews, conference and workshop proceedings, personal files). For vibration-exposed populations, qualifying papers were scored in terms of the provenance of their measuring instruments (adequacy of documentation, standardisation, reliability, criterion related and content validity). Similar criteria were applied to general proposals for whole diagnostic schemes, and evidence was collated on the test-retest reliability of symptom histories and clinical signs. RESULTS: In total, 23 relevant reports were identified concerning vibration-exposed populations--21 involving symptoms and 9 involving examination/diagnosis. Most of the instruments employed scored poorly in terms of methodological quality. The search also identified, from the wider literature, more than a dozen schemes directed at classifying ULDs, and 18 studies of test-retest reliability of symptoms and physical signs in the upper limb. Findings support the use of the standardised Nordic questionnaire for symptom inquiry and suggest that a range of physical signs can be elicited with reasonable between-observer agreement. Four classification schemes rated well in terms of content validity. One of these had excellent documentation, and one had been tested for repeatability, agreement with an external reference standard, and utility in distinguishing groups that differed in disability, prognosis and associated risk factors. CONCLUSIONS: Hitherto, most studies of ULDs in vibration-exposed populations have used custom specified diagnostic methods, poorly documented, and non-stringent in terms of standardisation and supporting evidence of reliability and/or validity. The broader literature contains several question sets and procedures that improve upon this, and offer scope in vibration-exposed populations to diagnose ULDs more systematically. PMID- 17909840 TI - Resting mechanomyography before and after resistance exercise. AB - A number of mechanisms have been proposed to explain the elevation in oxygen consumption following exercise. Biochemical processes that return muscle to its pre-exercise state do not account for all of the extra oxygen consumed after exercise (excess post-exercise oxygen consumption, EPOC). Muscle at rest after aerobic exercise produces mechanomyographic (MMG) activity of increased amplitude, compared to the pre-exercise state, which declines exponentially with the same time constant as EPOC. The purpose of this study was to determine how the resting MMG is affected by resistance exercise, and whether any change is related to oxygen consumption (VO(2)). Ten young male subjects (22.9 years) performed 30 min of resistance exercise consisting of one set of 10 repetitions at 50% 1-repetition maximum (1-RM) followed by five sets of eight repetitions at 75% of 1-RM for leg press and leg (knee) extension, with 1 min rest between sets. Oxygen consumption was measured by indirect calorimetry, MMG by an accelerometer placed over the rectus femoris, and surface electromyogram (EMG) with electrodes placed distal to the accelerometer. Recordings were made before exercise and for 5.5 h after exercise. MMG activity, expressed as mean absolute acceleration, was significantly elevated after exercise (P = 0.0006), as was EMG activity expressed as root-mean-square voltage (P = 0.03). MMG and VO(2) demonstrated exponential decay after exercise with similar time constants of 7.5 +/- 2.2 and 7.2 +/- 1.0 min, respectively. We conclude that resting muscle is more mechanically active following resistance exercise and that this may contribute to an elevated VO(2). PMID- 17909841 TI - The influence of crank length and cadence on mechanical efficiency in hand cycling. AB - The purpose of this study was to determine the effect of crank length and cadence on mechanical efficiency in hand cycling. Eight wheelchair dependent, high performance athletes completed four 4-min submaximal exercise bouts at a constant power output of 90 W over the different experimental conditions (crank length, pedal rate) using a sports hand bike (Draft, Godmanchester, UK). Two different crank lengths (180 and 220 mm) were tested at two different cadences (70 and 85 rev min(-1)) using the synchronous mode of cranking. Physiological measures of oxygen uptake (VO2) minute ventilation, blood lactate (B[La]), heart rate (HR), rate of perceived exertion (RPE) were recorded, gross (GE) and net (NE) efficiency were calculated. A two-way ANOVA with repeated measures was applied to determine the effects of crank length, cadence and their interaction on these physiological measures. Both GE and NE were significantly higher and V(O)(2) significantly lower for the 180 mm crank (P < 0.05). No significant main effect was found for cadence on the physiological measures (P > 0.05). Likewise, no interactions between crank length and pedal rate were found. There was however, a trend observed with HR and B[La] often lower with the 180 mm crank, indicating lower physiological stress. The RPE data supported this finding, with a tendency for lower ratings with the 180 mm crank (9 +/- 2 vs. 10 +/- 3). The short crank length when used at 85 rev min(-1) was found to be the most efficient (GE 21.4 +/ 3.1%). In conclusion, crank length has a significant effect on ME in hand cycling. A shorter crank length of 180 mm was found to be more efficient than the 220 mm, regardless of pedal rate during hand cycling. PMID- 17909843 TI - An improved method to compute the solute and water derangements of hyperglycaemia. AB - Evaluation and treatment of hyperglycaemic hyponatremia, being quantitatively inaccurate, is open to new advancements. We herein describe the improvement of previous calculations of glucose appearance (G(A)), solute and solvent changes. From G(A) we derive the predicted plasma sodium concentration (PNa(G)), assuming no change in total body water (TBW), but only water shift from cells to the extracellular space (ECV). This assumption is validated by the respective solute ratios (PCl/PNa) unchanged from normal values, as well as the ratios between actual and normal solute concentrations (PNa(1)/PNa(0), PCl(1)/PCl(0)), identical for all solutes. When the assumption is met, G(A) can be exactly calculated. When the ratios are different from normal, they indicate the presence of a mixed abnormality due to a loss either of sodium, or sodium and water. These are estimated by computing the difference between PNa(G) and the actual PNa measured (PNa(1)). PNa(1) approximately equal PNa(G) if TBW and Na are unchanged, PNa(1) < PNa(G) in the presence of prevalent Na depletion, PNa(1) > PNa(G )when volume depletion prevails. In the first circumstance the ECV expansion is exactly established by appropriate mathematical formulas, in the latter conditions either Na or volume depletion are empirically estimated with algebric expressions. These equations were validated on computer-simulated models, and applied to 49 subjects with plasma glucose concentration >15 mM/L. G(A) and PNa(G) were computed, and, with the same formulas used in computer-simulated experiments, we calculated water and Na deficits. The PNa measured after correction of hyperglycaemia was correctly predicted (R(2) = 0.63, P < 0.0001). This method provides a firm ground to select the correct equation to accurately estimate the initial conditions of hyperosmolar hyperglycaemia, significantly improving its quantitative correction. PMID- 17909842 TI - Cardiorespiratory responses during three repeated incremental exercise tests in sickle cell trait carriers. AB - This study investigated the cardioventilatory responses during heavy exercise in sickle cell trait carriers (SCTc) and subjects with normal hemoglobin (control group). Eight SCTc and six control subjects repeated three incremental exercise tests (Iet) separated by 10-min recoveries. Cardioventilatory parameters were analyzed at rest and during the first and third Iet. No significant difference in the ventilatory parameters [notably, maximal oxygen uptake (VO2max) and the ventilatory thresholds] was observed between the two groups. The time course of power output showed a significant difference between the first and third Iet from 80% of VO2max to VO2max (P < 0.05) in both groups. In conclusion, SCTc exhibited normal ventilatory responses during three successive Iet, which strongly suggests that this population, despite the presence of HbS in their red blood cells, is not limited during this type of aerobic exercise. PMID- 17909844 TI - Right ventricular function with hypoxic exercise: effects of sildenafil. AB - The effect of sildenafil on right ventricular contractility in hypoxic exercise is unknown, whereas reports have shown that sildenafil is associated with a smaller increase in pulmonary vascular resistance and right ventricular systolic pressure (RVSP) with exercise at high altitude. The present study evaluates the changes induced by controlled hypoxia on right ventricular pressure and performance with and without sildenafil administration. Tricuspid annular isovolumic acceleration (IVA) and annular velocities were measured in 14 healthy subjects at rest and after maximal exercise in a cross-over, double blind placebo controlled trial in three situations: normoxia, normobaric hypoxia with, and normobaric hypoxia without the administration of 100 mg sildenafil. RVSP, assessed by Doppler echocardiography, was determined from the peak tricuspid regurgitation pressure gradient. RVSP during rest increased from 26.9 +/- 2.3 mmHg in normoxia to 37.8 +/- 6.9 mmHg in hypoxia, p < 0.01; sildenafil administration reduced RVSP in hypoxia to 30.5 +/- 5.6, p < 0.01. Compared to normoxia at rest, IVA increased similarly with peak exercise in normoxia and hypoxia(sildenafil) (by 2.37 and 1.90 m/s(2), respectively), but the observed increase in IVA during exercise was smaller (0.86 m/s(2), p < 0.05) in hypoxia(placebo). Right ventricular contractility, as estimated by IVA at peak exercise is increased with the administration of sildenafil as compared to placebo, and is not different from the values seen during exercise in normoxia. This effect seems independent of the effect of sildenafil on RVSP. PMID- 17909845 TI - Hyperoxia improves 20 km cycling time trial performance by increasing muscle activation levels while perceived exertion stays the same. AB - Increasing inspiratory oxygen tension improves exercise performance. We tested the hypothesis that this is partly due to changes in muscle activation levels while perception of exertion remains unaltered. Eleven male subjects performed two 20-km cycling time-trials, one in hyperoxia (HI, FiO2 40%) and one in normoxia (NORM, FiO2 21%). Every 2 km we measured power output, heart rate, blood lactate, integrated vastus lateralis EMG activity (iEMG) and ratings of perceived exertion (RPE). Performance was improved on average by 5% in HI compared to NORM (P < 0.01). Changes in heart rate, plasma lactate concentration and RPE during the trials were similar. For the majority of the time-trials, power output was maintained in HI, but decreased progressively in NORM (P < 0.01) while it increased in both trials for the last kilometre (P < 0.0001). iEMG was proportional to power output and was significantly greater in HI than in NORM. iEMG activity increased significantly in the final kilometer of both trials (P < 0.001). This suggests that improved exercise performance in hyperoxia may be the result of increased muscle activation leading to greater power outputs. The finding of identical RPE, lactate and heart rate in both trials suggests that pacing strategies are altered to keep the actual and perceived exercise stress at a similar level between conditions. We suggest that a complex, intelligent system regulates exercise performance through the control of muscle activation levels in an integrative manner under conditions of normoxia and hyperoxia. PMID- 17909846 TI - Risk factor stratification after simultaneous liver and colorectal resection for synchronous colorectal metastasis. AB - BACKGROUND/AIM: This study was conducted to devise a prognostic model for patients undergoing simultaneous liver and colorectal resection. MATERIALS AND METHODS: A retrospective analysis was performed on 138 colorectal patients who underwent simultaneous liver and colorectal resection between September 1994 and September 2005. The primary endpoint of the study was overall survival. Three patients with positive liver resection margin were excluded from the analysis. RESULTS: At multivariate level, poor prognostic factors were liver resection margin < or =5 mm (P = 0.047; relative risk, 1.684; 95% CI = 1.010-2.809), CEA greater than 5 ng/ml (P = <0.001; relative risk, 2.507; 95% CI = 1.499-4.194), number of liver metastasis > 1 (P = <0.042; relative risk, 1.687; 95% CI = 1.020 2.789), and lymph node > or = 4 (P = <0.012; relative risk, 1.968; 95% CI = 1.158 3.347). The risk stratification grouping of the 135 patients was performed according to the following criteria: low risk group, 0-1 factor; intermediate risk group, 2 factors; high-risk group, 3-4 factors. Of 135 patients, 86 patients (63.0%) were categorized as low-risk group, 36 patients (26.6%) as intermediate risk group, and 14 patients (10.4%) as high-risk group. Median survival times for low, intermediate, high-risk groups were 68.0, 43.6 (95% CI, 24.7-62.4), and 23.5 months (95% CI, 9.4-31.5), respectively. The high-risk group demonstrated an approximately threefold (relative risk, 3.1; 95% CI, 1.6-6.0) increased risk of death. CONCLUSIONS: A simple risk factor stratification system was proposed to evaluate the chances of cure of patients after simultaneous resection of liver metastases and primary colorectal carcinoma. The risk factor stratification showed three groups with distinct survival. The risk stratification may help to predict patient survival after simultaneous liver and colorectal resection. This system needs further prospective validation. PMID- 17909847 TI - Pain after minimally invasive videoassisted and after minimally invasive open thyroidectomy--results of a prospective outcome study. AB - BACKGROUND: Substantial modifications in surgical treatment of thyroid disease have changed the postoperative management of thyroidectomized patients. The reduction of postoperative pain permit a short-stay surgery. MATERIALS AND METHODS: We have analyzed the patients treated in our Unit from July 2006 to December 2006, with minimally invasive cervicotomy and mini-invasive video assisted thyroidectomy. We have registered the postoperative pain applying an evaluation protocol numeric scale. The results were analyzed by t test. RESULTS: One hundred thirteen patients were divided in two groups: group A, minimally invasive cervicotomy (15 male and 46 female patients); group B, mini-invasive video-assisted thyroidectomy (9 male and 43 female patients). Upon returning to the ward, the pain scale group A vs B was 2.77 +/- 1.16 vs 2.5 +/- 0.762 (p = 0.22) .At 24 h after surgery, the pain scale in group A was 1.82 +/- 1.258 vs 1.031 +/- 0.8608 (p < 0.005). CONCLUSIONS: Both methods are safe, but mini invasive video-assisted thyroidectomy gives not only a better cosmetic result but a reduction of postoperative pain especially at 24 h. PMID- 17909848 TI - Release of cardiac troponin I from viable cardiomyocytes is mediated by integrin stimulation. AB - Elevated cardiac troponin-I (cTnI) levels have been demonstrated in serum of patients without acute coronary syndromes, potentially via a stretch-related process. We hypothesize that this cTnI release from viable cardiomyocytes is mediated by stimulation of stretch-responsive integrins. Cultured cardiomyocytes were treated with (1) Gly-Arg-Gly-Asp-Ser (GRGDS, n = 22) to stimulate integrins, (2) Ser-Asp-Gly-Arg-Gly (SDGRG, n = 8) that does not stimulate integrins, or (3) phosphate-buffered saline (control, n = 38). Cells and media were analyzed for intact cTnI, cTnI degradation products, and matrix metalloproteinase (MMP)-2. Cell viability was examined by assay of lactate dehydrogenase (LDH) activity and by nuclear staining with propidium iodide. GRGDS-induced integrin stimulation caused increased release of intact cTnI (9.6 +/- 3.0%) as compared to SDGRG treated cardiomyocytes (4.5 +/- 0.8%, p < 0.001) and control (3.0 +/- 3.4%, p < 0.001). LDH release from GRGDS-treated cardiomyocytes (15.9 +/- 3.8%) equalled that from controls (15.2 +/- 2.3%, p = n.s.), indicating that the GRGDS-induced release of cTnI is not due to cell necrosis. This result was confirmed by nuclear staining with propidium iodide. Integrin stimulation increased the intracellular and extracellular MMP2 activity as compared to controls (both p < 0.05). However, despite the ability of active MMP2 to degrade cTnI in vitro, integrin stimulation in cardiomyocytes was not associated with cTnI degradation. The present study demonstrates that intact cTnI can be released from viable cardiomyocytes by stimulation of stretch-responsive integrins. PMID- 17909849 TI - Bradykinin does not induce gap formation between human endothelial cells. AB - Generally, a formation of paracellular gaps is considered to be the main pathway for fluid passage across endothelia. A model substance for studies in vitro is the vasodilatory peptide bradykinin, which has important functions in inflammation and vascular fluid balance. The mechanisms by which it increases endothelial permeability are not as yet clearly defined. Paracellular gap formation was approached using atomic force microscopy (AFM) on human umbilical vein endothelial cells grown on permeable filter supports. To further distinguish between para- vs transcellular fluid passage, a standard permeability assay was modified by a rapid cooling protocol to specifically inhibit vesicular transport pathways. Cell layers stimulated with bradykinin (1 microM) did not show significant alterations at the cellular junctions. However, gap formation was easily detectable by AFM after addition of the Ca(2+)-ionophore ionomycin (1 microM), which was taken as a positive control for cellular contraction. At 37 degrees C, bradykinin enhanced fluorescein isothiocyanate-dextran permeability by 48 +/- 11%. This was blocked by rapid cooling of the sample, indicating a vesicular mechanism of fluid transport. Contrastingly, ionomycin-induced permeability (259 +/- 43%) persisted after cooling (230 +/- 44%), thereby confirming paracellular gap formation. Accordingly, endocytotic vesicle formation, as detected by fluorescence microscopy, was upregulated by 68 +/- 15% through bradykinin action, while ionomycin did not show a significant effect (7 +/- 26%). The combined results of both permeability and morphometric studies lead to the conclusion that bradykinin promotes transcellular fluid passage rather than increasing paracellular diffusion. PMID- 17909850 TI - The sodium-bicarbonate cotransporter NBCe1 supports glutamine efflux via SNAT3 (SLC38A3) co-expressed in Xenopus oocytes. AB - The glutamine transporter SNAT3 contributes to the glutamine fluxes in liver, kidney, and brain. We heterologously co-expressed SNAT3 with the electrogenic sodium-bicarbonate cotransporter NBCe1 in Xenopus laevis oocytes and measured cytosolic pH and membrane current in voltage clamp. Because of the increased buffer capacity contributed by the NBCe1 (Becker and Deitmer in J Biol Chem 279:28057-28062, 2004), we hypothesized that this may enhance the proton-coupled glutamine transport via SNAT3 in the presence of CO2/HCO3-. Addition and removal of glutamine activated not only SNAT3 but also NBCe1, as indicated by the increased membrane current. The NBCe1 current during glutamine removal was more than 50% larger than during glutamine addition, suggesting that NBCe1 enhances glutamine efflux rather than glutamine uptake. This was confirmed by radio labeled glutamine flux measurements; influx of glutamine was significantly decreased, whereas efflux of glutamine was increased when SNAT3 was co-expressed with NBCe1. A model is presented that attempts to explain the role of intracellular pH, bicarbonate transport, and buffering capacity mediated by NBCe1 for uptake and efflux of glutamine via SNAT3. PMID- 17909852 TI - Protein kinase C activation inhibits alpha1D L-type Ca channel: a single-channel analysis. AB - The recently reported alpha1D Ca channel in the heart is known to be regulated by protein kinase C (PKC) at the whole cell level and has been implicated in atrial fibrillation. The biophysical basis of this regulation at the single-channel level is not known. Therefore, the effect of PKC activation was studied on alpha1D Ca channel expressed in tsA201 cells using cell-attached configuration. Unitary currents were recorded in the presence of 70 mM Ba2+ as the charge carrier at room temperature. Under basal condition, channel activity was rare and infrequent; however, Bay K 8644 (1 microM) induced channel openings with a conductance of 22.3 pS. Single channel analysis of open and closed time distributions were best fitted with a single exponential. PKC activation by 4alpha-phorbol 12-myristate 13-acetate (PMA; 10 nM), a phorbol ester derivative, resulted in a decrease in open probability and increase in closed-time without any significant effect on the conductance of the alpha1D Ca channel. This is consistent with a decreased entry of alpha1D Ca channel into open states in the presence of PMA. PMA effects could not be reproduced by 4-alpha Phorbol, an inactive PMA analogue. These data show, for the first time, (1) the alpha1D Ca channel activity at the single-channel level and (2) the biophysical basis by which PKC activation inhibits the alpha1D Ca channel. The shortening of the open time and the lengthening of the closed-time constants and the increase in blank sweeps may explain the inhibition of the previously reported whole-cell alpha1D Ca current. Altogether, these data are essential for understanding the complex role of alpha1D Ca channel not only in physiological settings but also in pathological settings such as atrial fibrillation. PMID- 17909851 TI - Caloxins: a novel class of selective plasma membrane Ca2+ pump inhibitors obtained using biotechnology. AB - Plasma membrane Ca2+ pumps (PMCA) extrude cellular Ca2+ with a high affinity and hence play a major role in Ca2+ homeostasis and signaling. Caloxins (selective extracellular PMCA inhibitors) would aid in elucidating the physiology of PMCA. PMCA proteins have five extracellular domains (exdoms). Our hypotheses are: 1) peptides that bind selectively to each exdom can be invented by screening a random peptide library, and 2) a peptide can modulate PMCA activity by binding to one of the exdoms. The first caloxin 2a1, selected for binding exdom 2 was selective for PMCA (Ki=529 microM). It has been used to examine the physiological role of PMCA. PMCA isoforms are encoded by four genes. PMCA isoform expression differs in various cell types, with PMCA1 and 4 being the most widely distributed. There are differences between PMCA1-4 exdom 1 sequences, which may be exploited for inventing isoform selective caloxins. Using exdom 1 of PMCA4 as a target, modified screening procedures and mutagenesis led to the high-affinity caloxin 1c2 (Ki=2.3 microM for PMCA4). It is selective for PMCA4 over PMCA1, 2, or 3. We hope that caloxins can be used to discern the roles of individual PMCA isoforms in Ca2+ homeostasis and signaling. Caloxins may also become clinically useful in cardiovascular diseases, neurological disorders, retinopathy, cancer, and contraception. PMID- 17909853 TI - Consequences of prolonged total thermoneutral immersion on muscle performance and EMG activity. AB - We hypothesized that the changes in muscle temperature and interstitial pressure during thermoneutral immersion may affect the reflex adaptation of the motor drive during static contraction, assessed by the decrease in median frequency (MF) of electromyogram (EMG) power spectrum. Ten subjects were totally immersed for 6 h at 35 degrees C and repeated maximal voluntary contraction (MVC) and submaximal (60% MVC) leg extensions sustained until exhaustion. In vastus lateralis (VL) and soleus (SOL) muscles, the compound muscle potential evoked by muscle stimulation with single shocks (M-wave) was recorded at rest, and MF of surface EMG was calculated during 60% MVCs. We measured lactic acid and potassium venous blood concentrations and calculated plasma volume changes. Data were compared to those obtained in the same individuals exercising at 35 degrees C under dry conditions where the MF decrease during 60% MVCs was modest (-4 to-5%). During immersion, the rectal temperature remained stable, but the thigh and calf surface temperatures significantly increased. Lactic acid and potassium concentrations did not vary, but plasma volume decreased from the 180th min of immersion. The M-wave did not vary in VL but was prolonged in SOL from the 30th min of immersion. From the 220th min of immersion, the maximal MF decrease was majored in both muscles (-18 to -22%). Thus, compared to the dry condition, total body thermoneutral immersion enhances fatigue-induced EMG changes in leg muscles, perhaps through the activation of warm-sensitive muscle endings and/or the changes in interstitial pressure because of vasodilatation. PMID- 17909854 TI - Excess copper induces accumulation of hydrogen peroxide and increases lipid peroxidation and total activity of copper-zinc superoxide dismutase in roots of Elsholtzia haichowensis. AB - The effects of excess copper (Cu) on the accumulation of hydrogen peroxide (H2O2) and antioxidant enzyme activities in roots of the Cu accumulator Elsholtzia haichowensis Sun were investigated. Copper at 100 and 300 microM significantly increased the concentrations of malondialdehyde and H2O2, and the activities of catalase (E.C. 1.11.1.6), ascorbate peroxidase (E.C. 1.11.1.11), guaiacol peroxidase (GPOD, E.C. 1.11.1.7) and superoxide dismutase (SOD, E.C. 1.15.1.1). Isoenzyme pattern and inhibitor studies showed that, among SOD isoforms, only copper-zinc superoxide dismutase (CuZn-SOD) increased. Excess Cu greatly increased the accumulation of superoxide anion (O2 (.-)) and H2O2 in E. haichowensis roots. This study also provides the first cytochemical evidence of an accumulation of H2O2 in the root cell walls as a consequence of Cu treatments. Experiments with diphenyleneiodonium as an inhibitor of NADPH oxidase, 1,2 dihydroxybenzene-3,5-disulphonic acid as an O2 (.-) scavenger, and N-N diethyldithiocarbamate as an inhibitor of SOD showed that the source of H2O2 in the cell walls could partially be NADPH oxidase. The enzyme can use cytosolic NADPH to produce O2 (.-), which rapidly dismutates to H2O2 by SOD. Apoplastic GPOD and CuZn-SOD activities were induced in roots of E. haichowensis with 100 microM Cu suggesting that these two antioxidant enzymes may be responsible for H2O2 accumulation in the root apoplast. PMID- 17909855 TI - Overexpression of CYP710A1 and CYP710A4 in transgenic Arabidopsis plants increases the level of stigmasterol at the expense of sitosterol. AB - Sitosterol and stigmasterol are major sterols in vascular plants. An altered stigmasterol:sitosterol ratio has been proposed to influence the properties of cell membranes, particularly in relation to various stresses, but biosynthesis of stigmasterol is poorly understood. Recently, however, Morikawa et al. (Plant Cell 18:1008-1022, 2006) showed in Arabidopsis thaliana that synthesis of stigmasterol and brassicasterol is catalyzed by two separate sterol C-22 desaturases, encoded by the genes CYP710A1 and CYP710A2, respectively. The proteins belong to a small cytochrome P450 subfamily having four members, denoted by CYP710A1-A4, and are related to the yeast sterol C-22 desaturase Erg5p acting in ergosterol synthesis. Here, we report on our parallel investigation of the Arabidopsis CYP710A family. To elucidate the function of CYP710A proteins, transgenic Arabidopsis plants were generated overexpressing CYP710A1 and CYP710A4. Compared to wild-type plants, both types of transformant displayed a normal phenotype, but contained increased levels of free stigmasterol and a concomitant decrease in the level of free sitosterol. CYP710A1 transformants also displayed higher levels of esterified forms of stigmasterol, cholesterol, 24-methylcholesterol and isofucosterol. The results confirm the findings of Morikawa et al. (Plant Cell 18:1008-1022, 2006) regarding the function of CYP710A1 in stigmasterol synthesis, and show that CYP710A4 also has this capacity. Furthermore, our results suggest that an increased stigmasterol level alone is sufficient to stimulate esterification of other major sterols. PMID- 17909856 TI - Varicella-zoster virus infection induces the secretion of interleukin-8. AB - Interleukin-8 (IL-8) is an important mediator in neutrophil-mediated acute inflammation but has also a wide range of actions on various cells types. We demonstrated that infection of melanoma cells and fibroblasts with cell associated varicella-zoster virus (VZV) and infection of a T cell line with cell free VZV resulted in an induction of IL-8 secretion in vitro. The inhibition of the VZV replication with a drug interfering with its DNA replication had no effect on the IL-8 release. Since the IL-8 promoter contains binding sites for NF kappaB and AP-1, melanoma cells and the T cell line were treated with inhibitors of NF-kappaB, JNK/SAPK or p38/MAPK prior to infection. In melanoma cells, the JNK/SAPK pathway was shown to be important for the IL-8 secretion during the VZV replication, whereas in the T cell line, not only the JNK/SAPK but also the p38/MAPK pathways were required for IL-8 secretion. The neutralisation of the IL 8 bioactivity had no significant consequence on the VZV replication, suggesting that IL-8 acts neither as a proviral nor as an antiviral cytokine during the VZV replication in vitro. PMID- 17909858 TI - Expression, localization, and inducibility by bile acids of hepatobiliary transporters in the new polarized rat hepatic cell lines, Can 3-1 and Can 10. AB - Sinusoidal and apical transporters are responsible for the uptake and biliary elimination of many compounds by hepatocytes. Few in vitro models are however available for analyzing such functions. The expression and bile-acid inducibility of 13 transporters and two nuclear receptors were investigated in the new rat polarized lines, Can 3-1 and Can 10, and in their unpolarized parent, Fao. The relative abundance of mRNA, the protein level, and their localization were examined by real-time quantitative PCR, Western blotting, immunofluorescence, and confocal microscopy. Compared with rat liver, mRNA levels of Fao cells were: negligible for Bsep/Abcb11; lower for the uptake transporters Ntcp and Oatps; similar for SHP, FXR, and Bcrp/Abcg2; and higher (four-fold to 160-fold) for the efflux pumps Mdr1b/Abcb1b, Mdr2/Abcb4, Mrp1/Abcc1, Mrp2/Abcc2, Mrp3/Abcc3, Abcg5, and Abcg8. This profile was mostly maintained (and improved for Bsep) in Can 10. Some transporters were less well expressed in Can 3-1. In both lines, sinusoidal (Ntcp, Mrp3) and canalicular transporters (Mdr-P-glycoproteins detected with C219 antibody, Mrp2) were localized at their correct poles. Bile-acid effects on polarity and mRNA levels of transporters were analyzed after a 6-day treatment with 50 microM taurocholic, chenodeoxycholic (CDCA), or ursodeoxycholic acid (UDCA). No polarization of Fao cells was induced; Can 10 and Can 3-1 polarity was maintained. CDCA and UDCA induced marked enhancement of the volume of Can 10 bile canaliculi. CDCA upregulated Bsep, Mdr2, SHP, Mdr1b, and Oatp2/1a4 in Can 10 (two to seven-fold) and in Fao cells. Thus, Can 10 constitutes an attractive polarized model for studying vectorial hepatobiliary transport of endogenous and xenobiotic cholephilic compounds. PMID- 17909857 TI - Enhancement of dendritic cell-tumor fusion vaccine potency by indoleamine-pyrrole 2,3-dioxygenase inhibitor, 1-MT. AB - PURPOSE: Dendritic cell (DC)-based cancer vaccines are currently being evaluated as novel anti-tumor vaccination strategies, but in some cases, they are demonstrated to have poor clinical efficacies than anticipated. A potential reason is immune tolerance due to the immunosuppressive enzyme, indoleamine pyrrole 2,3-dioxygenase (IDO). The aim of this study was to determine whether blocking the activity of IDO might improve the anti-tumor efficacy of DC/Lewis lung carcinoma (LLC) fusion vaccine applied to the mouse LLC model. METHODS: To prepare the DC/LLC fusion vaccine, DCs were fused with LLC using polyethylene glycol (PEG) as described. The IDO expression in the DC/LLC fusion vaccine and in the vaccinated mice was detected by western blot (WB) and/or immunohistochemical (IHC) analysis. This fusion vaccine, as a single agent or in combination with 1 methyl-tryptophan (1-MT, an IDO inhibitor), was administered to LLC mice. The anti-tumor efficacy in different treatment was determined by regular observation of tumor development and the level of splenic cytotoxic T lymphocyte (CTL) response, which was examined by lactate dehydrogenase (LDH) release. RESULTS: In the LLC mice, we observed that IDO-positive cells were extensively accumulated in tumor draining lymph nodes (TDLNs). Furthermore, WB and IHC analysis results showed that vaccination with fusion DC/LLC cells alone caused significant up regulation of IDO in spleens. 1-MT enhanced the anti-tumor efficacy elicited by DC/LLC fusion vaccine via delaying the tumor development and inducing stronger splenic CTL responses. CONCLUSIONS: Our results indicate an IDO-mediated immunosuppressive mechanism might be involved in weakening the anti-tumor efficacy elicited by DC/LLC fusion vaccine, and specific inhibition of IDO activity might be required for development of cancer vaccines. PMID- 17909860 TI - Simultaneous organics removal and bio-electrochemical denitrification in microbial fuel cells. AB - Simultaneous organics removal and bio-electrochemical denitrification using a microbial fuel cell (MFC) reactor were investigated in this study. The electrons produced as a result of the microbial oxidation of glucose in the anodic chamber were transferred to the anode, which then flowed to the cathode in the cathodic chamber through a wire, where microorganisms used the transferred electrons to reduce the nitrate. The highest power output obtained on the MFCs was 1.7 mW/m(2) at a current density of 15 mA/m(2). The maximum volumetric nitrate removal rate was 0.084 mg NO(3)(-)-N cm(-2) (electrode surface area) day(-1). The coulombic efficiency was about 7%, which demonstrated that a substantial fraction of substrate was lost without current generation. PMID- 17909859 TI - Identification of cellular isoform of oviduct-specific glycoprotein: role in oviduct tissue remodeling? AB - The oviduct is known to secrete mucins (MUC1 and MUC9) under the influence of ovarian steroids. The secreted form of MUC1 binds gametes in the oviduct, whereas the cellular form seen in breast cancers has been implicated in cell adhesion and morphogenesis. The secreted MUC9 or oviduct-specific glycoprotein (OGP), in addition to being a mucin, belongs to family 18 glycosylhydrolases and is known to bind gametes and embryos in the oviduct. Studies in our laboratory have identified non-muscle myosin IIA (involved in cell shape, polarity, and morphogenesis) as the protein partner to OGP in gametes. In view of the crucial role of the cortical cytoskeleton in the selective internalization of tight junctions (TJs) /adherent junctions (AJs) or apical junctional complex (AJC) in simple epithelial cells during tissue remodeling, the present study has been undertaken to evaluate the existence of a cellular form of OGP in oviductal tissue, which itself undergoes cyclic tissue remodeling. In silico analysis of the deduced amino-acid sequence of OGP has revealed the presence of several conserved motifs; these imply that OGP is a component of multi-protein complexes such as TJs. Corroborative immunoelectron-microscopic analysis in peri-ovulatory oviduct epithelia in the bonnet monkey has revealed the presence of OGP at the TJ. Co-localization studies of OGP and cadherin demonstrate that, whereas OGP is localized at the tonofilaments of the TJs, cadherin is localized at the intercellular space of the AJ. The possible role of OGP in oviductal tissue remodeling is discussed in light of the present findings and those reported in the literature. PMID- 17909861 TI - Modeling for the optimal biodegradation of toxic wastewater in a discontinuous reactor. AB - The degradation of toxic compounds in Sequencing Batch Reactors (SBRs) poses inhibition problems. Time Optimal Control (TOC) methods may be used to avoid such inhibition thus exploiting the maximum capabilities of this class of reactors. Biomass and substrate online measurements, however, are usually unavailable for wastewater applications, so TOC must use only related variables as dissolved oxygen and volume. Although the standard mathematical model to describe the reaction phase of SBRs is good enough for explaining its general behavior in uncontrolled batch mode, better details are needed to model its dynamics when the reactor operates near the maximum degradation rate zone, as when TOC is used. In this paper two improvements to the model are suggested: to include the sensor delay effects and to modify the classical Haldane curve in a piecewise manner. These modifications offer a good solution for a reasonable complexification tradeoff. Additionally, a new way to look at the Haldane K-parameters (micro(o),K(I),K(S)) is described, the S-parameters (micro*,S*,S(m)). These parameters do have a clear physical meaning and, unlike the K-parameters, allow for the statistical treatment to find a single model to fit data from multiple experiments. PMID- 17909862 TI - Self-reported experience and outcomes of care among stomach cancer patients at a median follow-up time of 27 months from diagnosis. AB - GOALS OF WORK: We aimed to identify clinical experiences associated with outcomes of care among stomach cancer patients. MATERIALS AND METHODS: Four hundred thirty two patients who had a diagnosis of stage I-III stomach cancer from 2001 through 2002 from two hospitals in South Korea responded to a survey questionnaire including sociodemographic and clinical data, information about care experiences, satisfaction with care, and quality of life (QOL). MAIN RESULTS: Involvement in decision making [adjusted odds ratio (aOR) = 1.81; 95% confidence interval (CI), 1.13 to 2.89] and reflection of patients' opinions in treatment decisions (aOR = 2.54; 95% CI, 1.65 to 3.93) were associated with decision satisfaction. The factors associated with willingness to choose the same treatment over again were involvement in decision making (aOR = 2.37; 95% CI, 1.53 to 3.68) and no treatment toxicity (aOR = 0.50; 95% CI, 0.29 to 0.87). Involvement in decision making, reflection of patients' opinions in treatment decisions, and treatment toxicity were associated with some functioning subscales of QOL (p < 0.05). Regular follow-up, however, was associated with poor social functioning. CONCLUSIONS: Quality improvement efforts for stomach cancer patients should focus not only on the quality of primary tumor therapy but also on how patients experience their care, such as patient-centered decision making, experience of treatment toxicity, and regular follow-up. PMID- 17909863 TI - A cost calculation model for specialist palliative care for patients with non small cell lung cancer in a tertiary centre. AB - GOALS OF WORK: Five-year survival of patients with non-small cell lung cancer (NSCLC) is below 15%. Therefore, an early integration of palliative care according to the 2002 WHO definition is indispensable. In this paper, we describe methodical and financial aspects of prospective pricing of palliative care within a concept of integrated care for patients with NSCLC in Germany. MATERIALS AND METHODS: Four structures of palliative care services were defined (hospital support, home care, day care and in-patient care). Prospectively, resource use was estimated, using real cost data from the finance department of the University Hospital. Resource use was forecasted on the basis of operating experience, data of the national core documentation of palliative care patients and recommendations from the European Commission. RESULTS: Expected average hospital support team services were priced at 483 euros and budgeted for 10% (stage 1) to 90% (stage 4) of patients. Home care (60 visits, 4,573 euros) and day-care (5 visits) services were budgeted for between 5% (stage 1) and 30% (stage 4). The resulting prospective reimbursements range from 393 euros (stage 1) to 2,503 euros (stage 4). In-patient care was excluded from the prospective payments and reimbursed separately. CONCLUSIONS: For the first time, global reimbursements covering palliative care hospital support, home care and day care for patients with NSCLC were prospectively calculated and successfully negotiated. The contractual specification of palliative care services may contribute to transparency and quality in cancer care. PMID- 17909864 TI - Quality of analgesic treatment in patients with advanced prostate cancer: do we do a better job now? The Swiss Group for Clinical Cancer Research (SAKK) experience. AB - GOALS OF WORK: The aim of this study was to evaluate pain intensity and the application of the WHO guidelines for cancer pain treatment in patients with prostate cancer treated at Swiss cancer centers. MATERIALS AND METHODS: We analyzed a series of five multicenter phase II clinical trials which examined the palliative effect of different chemotherapies in patients with advanced hormone refractory prostate carcinoma. Of 170 patients, 1,018 visits were evaluable for our purpose, including ratings of pain intensity by patients and prescribed analgesics. MAIN RESULTS: No or mild pain was indicated by patients in 36 to 55% of the visits, more than mild pain in 30 to 46%. In 21% of the visits, the WHO pain treatment criteria (treatment according to one of the three steps; oral, rectal or transdermal application of the main dose; administration on a regular schedule) were fulfilled, and the Cleeland index was positive according to all recommendations. In 6% of the visits, neither the WHO criteria were fulfilled nor was the Cleeland index positive. This indicates insufficient pain treatment not following the WHO guidelines and that the prescribed analgesics were not sufficiently potent for the rated pain intensity. CONCLUSIONS: In this selective Swiss sample, the standard of analgesic treatment is high. However, there is still scope for improvement. This cannot solely be solved by improving the knowledge of the physicians. Programs to change the patients' attitude towards cancer pain, training to improve the physicians' communication skills, and institutional changes may be promising strategies. PMID- 17909866 TI - Synthesis and characterization of new copper thiosemicarbazone complexes with an ONNS quadridentate system: cell growth inhibition, S-phase cell cycle arrest and proapoptotic activities on cisplatin-resistant neuroblastoma cells. AB - Two new copper thiosemicarbazone complexes with an ONNS quadridentate system were synthesized and evaluated for anticancer activity on cisplatin-resistant neuroblastoma cells. Among these two copper complexes, the substituted 8 hydroxyquinoline-2-carboxaldehyde-4,4-dimethyl-3-thiosemicarbazide (CuHQDMTS) exhibited stronger cell growth inhibition activity than the unsubstituted copper 8-hydroxyquinoline-2-carboxaldehyde thiosemicarbazide complex (CuHQTS). Both CuHQTS and CuHQDMTS showed dose-dependent cell growth inhibition, cell cycle arrest and apoptosis induction activities on the SK-N-DZ neuroblastoma cells. Increased expression of p53 protein molecules was detected in the SK-N-DZ cells treated with CuHQTS. The data obtained in this study suggest that CuHQDMTS and CuHQTS hold potential as new, effective drugs for treatment of refractory neuroblastoma in children. PMID- 17909865 TI - Retrospective chart review of severe infusion reactions with rituximab, cetuximab, and bevacizumab in community oncology practices: assessment of clinical consequences. AB - GOALS OF WORK: Monoclonal antibody (MoAb) treatments can result in severe infusion reactions. Managing infusion reactions in the outpatient setting introduces clinical and resource challenges for patients and providers, but there is little information regarding prevention, management, or outcomes of severe infusion reactions. This study represents one of the first attempts to describe the clinical consequences of severe infusion reactions associated with MoAb treatment. MATERIALS AND METHODS: Clinic staff identified adults treated with rituximab, cetuximab, or bevacizumab who experienced a grade 3 or higher (severe) infusion reaction. Chart reviews from 19 oncology practice sites across the USA captured patient demographics, infusion reaction management procedures, and clinical outcomes. MAIN RESULTS: With an average age of 62 years, the sample comprised of 76 patients who experienced a severe infusion reaction while receiving rituximab (n = 47), cetuximab (n = 24), and bevacizumab (n = 5). The most common pretreatment medications were acetaminophen and antihistamine in the rituximab group and corticosteroids (42%) in the cetuximab group. All cetuximab and the majority of rituximab severe infusion reactions occurred during the first cycle of therapy. Postinfusion reaction management typically included corticosteroids, oxygen, and intravenous fluids. Overall, 22% were hospitalized for a mean of 4 days (range = 2.0 to 6.0 days). Permanent discontinuation of MoAb therapy occurred after the majority of cetuximab (79 to 100%) related severe infusion reactions. CONCLUSIONS: Severe infusion reactions are intensive events that present a serious challenge to patients and oncology practices. Efforts to prevent or reduce such reactions could be of great benefit. PMID- 17909867 TI - Occurrence and distribution of sequence types among Legionella pneumophila strains isolated from patients in Germany: common features and differences to other regions of the world. AB - A total of 105 unrelated clinical isolates of Legionella pneumophila were randomly selected from the German National Legionella strain collection and typed by monoclonal antibody (MAb) subgrouping and a seven-gene locus sequence-based typing (SBT) scheme. According to the case definitions of the European Working Group for Legionella Infections, 19 of the isolates tested were travel associated, 38 were community-acquired and 48 were of nosocomial origin. Eighty four of these strains belonged to serogroup 1, 20 belonged to other serogroups, and one isolate could not be serogrouped. The majority of strains among the travel-associated and community-acquired cases were MAb3-1-positive. The most common sequence type (1, 4, 3, 1, 1, 1, 1) was found in 20 isolates in 11 cities; other allelic profiles also found in Europe (2, 3, 9, 10, 2, 1, 6), (1, 3, 9, 10, 2, 1, 6), (2, 6, 17, 14, 13, 11, 11) and (3, 4, 1, 1, 1, 9, 1) were detected among the German isolates but at a low frequency. In contrast, some SBT are unique to Germany, including (3, 4, 1, 3, 35, 9, 11), which was found among five isolates from patients in Berlin. In concordance with European data, a significant portion of the L. pneumophila strains isolated from patients in Germany belong to clones that occur throughout the world and which are responsible for the majority of clinical cases. PMID- 17909868 TI - Theoretical study of Beloussov's hyper-restoration hypothesis for mechanical regulation of morphogenesis. AB - Computational models were used to explore the idea that morphogenesis is regulated, in part, by feedback from mechanical stress according to Beloussov's hyper-restoration (HR) hypothesis. According to this hypothesis, active tissue responses to stress perturbations tend to restore, but overshoot, the original (target) stress. To capture this behavior, the rate of growth or contraction is assumed to depend on the difference between the current and target stresses. Stress overshoot is obtained by letting the target stress change at a rate proportional to the same stress difference. The feasibility of the HR hypothesis is illustrated by models for stretching of epithelia, cylindrical bending of plates, invagination of cylindrical and spherical shells, and early amphibian development. In each case, an initial perturbation leads to an active mechanical response that changes the form of the tissue. The results show that some morphogenetic processes can be entirely self-driven by HR responses once they are initiated (possibly by genetic activity). Other processes, however, may require secondary mechanisms or perturbations to proceed to completion. PMID- 17909870 TI - Targeted disruption of homoserine dehydrogenase gene and its effect on cephamycin C production in Streptomyces clavuligerus. AB - The aspartate pathway of Streptomyces clavuligerus is an important primary metabolic pathway which provides substrates for beta-lactam synthesis. In this study, the hom gene which encodes homoserine dehydrogenase was cloned from the cephamycin C producer S. clavuligerus NRRL 3585 and characterized. The fully sequenced open reading frame encodes 433 amino acids with a deduced M (r) of 44.9 kDa. The gene was heterologously expressed in the auxotroph mutant Escherichia coli CGSC 5075 and the recombinant protein was purified. The cloned gene was used to construct a plasmid containing a hom disruption cassette which was then transformed into S. clavuligerus. A hom mutant of S. clavuligerus was obtained by insertional inactivation via double crossover, and the effect of hom gene disruption on cephamycin C yield was investigated by comparing antibiotic levels in culture broths of this mutant and in the parental strain. Disruption of hom gene resulted in up to 4.3-fold and twofold increases in intracellular free L lysine concentration and specific cephamycin C production, respectively, during stationary phase in chemically defined medium. PMID- 17909869 TI - Antifouling activity of sessile bacilli derived from marine surfaces. AB - Marine biofilms are a virtually untapped source of bioactive molecules that may find application as novel antifoulants in the marine paint industry. This study aimed at determining the potential of marine biofilm bacteria to produce novel biomolecules with potential application as natural antifoulants. Nine representative strains were isolated from a range of surfaces and were grown in YEB medium and harvested during the late exponential growth phase. Bacterial biomass and spent culture medium were extracted with ethanol and ethyl acetate, respectively. Extracts were assayed for their antifouling activity using two tests: (1) antimicrobial well diffusion test against a common fouling bacterium, Halomonas marina, and (2) anti-crustacean activity test using Artemia salina. Our results showed that none of the ethanolic extracts (bacterial biomass) were active in either test. In contrast, most of the organic extracts had antimicrobial activity (88%) and were toxic towards A. salina (67%). Sequencing of full 16 S ribosomal DNA analysis showed that the isolates were related to Bacillus mojavensis and Bacillus firmus. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) profiling of ethyl acetate extracts of culture supernatants showed that these species produce the bioactive lipopeptides surfactin A, mycosubtilin and bacillomycin D. PMID- 17909871 TI - Acetate-mediated pH-stat fed-batch cultivation of transconjugant Enterobacter sp. BL-2S over-expressing glmS gene for excretive production of microbial polyglucosamine PGB-1. AB - A unique cationic polyglucosamine biopolymer PGB-1 comprising more than 95% D glucosamine was excretively produced from a new bacterial strain Enterobacter sp. BL-2 under acetate-mediated culture conditions. Since the biopolymer PGB-1 could be synthesized from the UDP-N-acetylglucosamine monomer derived from the hexosamine pathway, three glmS, glmM, and glmU genes in the hexosamine pathway were cloned from Enterobacter sp. BL-2, and their molecular structures were elucidated. The cloned glmS, glmM, and glmU genes were reintroduced into the parent strain Enterobacter sp. BL-2 through a conjugative transformation for the overproduction of the biopolymer PGB-1. The biopolymer production increased 1.5 fold in the transconjugant Enterobacter sp. BL-2S over-expressing the first-step glmS gene encoding glucosamine-6-phosphate synthase. The transconjugant Enterobacter sp. BL-2S was cultivated pH-stat fed-batch widely, while intermittently feeding an acetate solution to maintain a constant pH level of 8.0 for 72 h, resulting in 1.15 g/L of the extracellular polyglucosamine biopolymer PGB-1. PMID- 17909872 TI - Stability and activity of lipase in subcritical 1,1,1,2-tetrafluoroethane (R134a). AB - The stability and activity of commercial immobilized lipase from Candida antarctica (Novozym 435) in subcritical 1,1,1,2-tetrafluoroethane (R134a) was investigated. The esterification of oleic acid with glycerol was studied as a model reaction in subcritical R134a and in solvent-free conditions. The results indicated that subcritical R134a treatment led to significant increase of activity of Novozym 435, and a maximum residual activity of 300% was measured at 4 MPa, 30 degrees C after 7 h incubation. No deactivation of Novozym 435 treated with subcritical R134a under different operation factors (pressure 2-8 MPa, temperature 30-60 degrees C, incubation time 1-12 h, water content 1:1, 1:2, 1:5 enzyme/water, depressurization rate 4 MPa/1 min, 4 MPa/30 min, 4 MPa/90 min) was observed. While the initial reaction rate was high in subcritical R134a, higher conversion was obtained in solvent-free conditions. Though the apparent conversion of the reaction is lower in subcritical R134a, it is more practicable, especially at low enzyme concentrations desired at commercial scales. PMID- 17909873 TI - Neuroanatomical correlates of processing in visual and visuospatial working memory. AB - Working memory is traditionally seen as being organised in a modular way with a central executive orchestrating at least two slave systems (phonological loop and visuospatial sketch pad). Neuroanatomical correlates of the visual and visuospatial subsystems and the central executive are discussed in this article. A series of experiments are presented yielding evidence for a differentiation into active and passive processing in working memory as well as their neuroanatomical correlates in the prefrontal cortex. Data, yielding evidence for an interaction and separation of visual and visuospatial working memory are presented and discussed. Further results are presented which suggest a convergence of these two systems with increasing working memory demands. The discussed findings will give new insight in the organisation of visual and visuospatial working memory on the anatomical level. PMID- 17909874 TI - Retinal safety of a new fluoroquinolone, pradofloxacin, in cats: assessment with electroretinography. AB - PURPOSE: To investigate the safety of a new fluoroquinolone, pradofloxacin, on the cat retina using electroretinogram. METHODS: Ganzfeld ERGs were recorded in 40 cats treated orally for 23 days in 4 groups: CTRL (n = 9): placebo-vehicle; PRADO30 (n = 10): pradofloxacin 30 mg/kg/day; PRADO50 (n = 14): pradofloxacin 50 mg/kg/day; and ENRO30 (n = 7): enrofloxacin at toxic doses of 30 mg/kg/day. ERG was performed before treatment and once weekly during the treatment period. An extended ISCEV protocol with addition of 8 steps of increasing luminance in dark adapted condition was carried out to assess: V (max) (saturated scotopic b-wave amplitude) and k (luminance inducing V (max)/2). OCT and retinal histological changes were also investigated. RESULTS: Pradofloxacin showed no effects in respect to rod b-wave, V (max), k and maximum scotopic a-wave (P > 0.05). Oscillatory potentials, cone ERG and flicker were also unaltered (P > 0.05). Rod b-wave was undetectable after treatment in ENRO30 group, V (max) was reduced to 10.5% of the baseline (P < 0.05), accompanied by an increase of k by 1 log cd s/m(2) (P < 0.05). Oscillatory potentials, cone b-wave amplitude and 30 Hz flicker amplitude were reduced to 8.3%, 58.9% and 37.4% of the baseline, respectively (P < 0.05). Effects were also seen in OCT and retinal histology starting within one week after the start of treatment and thereafter remaining stable. CONCLUSION: Pradofloxacin at 6 and 10 times the recommended doses was shown to have no retinal toxic effects in cats, neither on rod or cone function with ERG. PMID- 17909875 TI - An artificial reflex improves the perturbation-resistance of a human walking simulator. AB - Most walking assist systems reported are not available for real-world environment, where frequent perturbations are caused by slips, uneven terrain, slopes and obstacles. On the other hand, it is evident that human beings cope with those perturbations, especially when the perturbations cannot be predicted or perceived in advance, with reflexes, which cause relatively fixed muscular responsive patterns to perturbations unconsciously within a short period of time ranging from several 10 to 200 ms. Our ultimate goal is to realize artificial reflexes in real-world walking support systems for those paralyzed people, whose afferent and efferent neural pathways are usually weakened, so that the reflexive system is also impaired to a certain degree. This goal needs both qualitative and quantitative understanding of human reflexive mechanism during walking. However, except for some hypotheses about the underlying neural mechanisms of the reflexes during walking, there is no widely accepted unified theory, nor are there clear experimental results that could be directly quoted in the disciplines of physiology and motor control. Our approach includes (1) acquiring muscle activity profiles during normal walking and slip-perturbed walking by recording and processing Electromyographic (EMG) signals of several walking-related muscles, in human gait experiments; (2) developing a central-pattern-generator (CPG) based neuro-musculo-skeletal simulation model; (3) comparing joint trajectories of the simulation model with those of a human subject during normal walking to verify the simulation model's conformity with human walking; (4) using muscle activity profiles of reflexive responses to slip-perturbation during walking to construct a rapid responding pathway. The results showed that, (1) The simulation model could show behavior resembling that of normal human walking; (2) in the case of occurrence of slip-perturbation, the rapid responding pathway could improve the perturbation-resistance and maintain the balance for the walking; (3) using the simulation model, several hypotheses on underlying neuro-mechanism were investigated. These reveal the possibility to realize the artificial reflex for the paralyzed people. PMID- 17909876 TI - Entropy improvement by the Temporal-Window method for alternating and non alternating 3D wavelet transform over angiographies. AB - The three-dimensional wavelet transform (3D-WT) has been proposed for volumetric data coding, since it can provide lossless coding and top-quality reconstruction: two key features highly relevant to medical imaging applications. In this paper, we present experimental results for four new algorithms based on the Classic 3D WT. The proposed algorithms are capable of obtaining the wavelet coefficients after the spatial and, mainly, the temporal decomposition processes, reducing most redundancies in the video sequence and getting lower entropy values than the Classic algorithm. The new algorithms are based on the Temporal-Window method for carrying out the temporal decomposition. We have conducted a set of experimental evaluations for a representative data set of a modality of intrinsically volumetric medical imaging: angiography sequences. PMID- 17909877 TI - Genetic studies of diabetes following the advent of the genome-wide association study: where do we go from here? PMID- 17909878 TI - Cholesterol, diabetes and major cardiovascular diseases in the Asia-Pacific region. AB - AIMS/HYPOTHESIS: The aim of this study was to assess the association between total cholesterol and major cardiovascular diseases among persons with and without diabetes in the Asia-Pacific region. METHODS: We used data on individual participants in 30 cohort studies from the Asia-Pacific region to compute the hazards ratios and 95% CIs for participants with and without diabetes at baseline, using Cox proportional models. Analyses were stratified by sex and region (Asia vs Australia or New Zealand) and adjusted for age. Repeat measurements of total cholesterol were used to adjust for regression dilution bias. RESULTS: The analysis included 333,533 individuals (6.3% with diabetes at baseline) who experienced 6,074 fatal and non-fatal cardiovascular events over a median follow-up period of 4.0 years. Total cholesterol was positively associated with coronary heart disease (CHD) and ischaemic stroke, and negatively with haemorrhagic stroke in a continuous, log-linear fashion, similarly among participants with and without diabetes. Each 1 mmol/l increase above the 'usual' level for total cholesterol was associated with a 41% (95% CI 23-63%) and 42% (95% CI 35-50%) greater risk of CHD among participants with and without diabetes. The corresponding values for ischaemic stroke were 23% (95% CI 0-52%) and 31% (95% CI 20-44%), respectively. These results were broadly consistent for sex, age and region. CONCLUSIONS/INTERPRETATION: Total cholesterol is associated with similarly increased risks of cardiovascular events in people with and without diabetes. While abnormal levels of other lipid fractions are frequently observed in people with diabetes, these data support aggressive lowering of total cholesterol and LDL-cholesterol levels for prevention of cardiovascular events. PMID- 17909879 TI - Association studies of ALOX5 and bone mineral density in healthy adults. AB - Animal studies suggest that arachidonate 5-lipoxygenase (encoded by ALOX5) may be a genetic determinant of bone mineral density. We tested this hypothesis in a sample of healthy men and women and did not find consistent evidence for an association between variation in this gene and either lumbar spine or femoral neck BMD. INTRODUCTION: Phenotypic variation in bone mineral density (BMD) among healthy adults is influenced by both genetic and environmental factors. A recent mouse study implicated ALOX5, which encodes arachidonate 5-lipoxygenase, as a contributing factor to areal BMD (aBMD). METHODS: Fifteen single nucleotide polymorphisms (SNPs) distributed throughout ALOX5 were genotyped in three healthy groups: 1,688 European American, premenopausal sisters, 512 African American premenopausal sisters and 715 European American brothers. Statistical analyses were performed in the three groups to test for association between these SNPs and femoral neck and lumbar spine aBMD. RESULTS: Significant (p < or = 0.05) evidence of association was observed with three of the SNPs. However, despite the linkage disequilibrium between SNPs, adjacent SNPs did not provide statistical evidence of association in any of the three study groups. CONCLUSIONS: These data do not provide consistent evidence of association between genomic variation in ALOX5 and clinical variability in aBMD in healthy subjects. PMID- 17909880 TI - A giant sigmoid diverticulum presenting as an upper abdominal mass. AB - AIM: We report a case of a giant diverticulum of the sigmoid colon presenting as a mass in the left upper quadrant of the abdomen in an elderly man. METHODS: This report highlights a rare complication of diverticular disease. At operation, the giant cyst was situated in the supracolic compartment of the abdomen. The management options and a review of the literature are presented. CONCLUSION: CT scan is the investigation of choice. Early surgical intervention is important in order to reduce the risk of perforation. PMID- 17909881 TI - Spatial and temporal trends of mercury concentrations in young-of-the-year spottail shiners (Notropis hudsonius) in the St. Lawrence River at Cornwall, ON. AB - The St. Lawrence River at Cornwall, Ontario is an "Area of Concern" because of mercury (Hg) biomagnification from bottom sediments. To assess the spatial and temporal distribution of Hg in the food web, young-of-the-year (YOY) spottail shiners (Notropis hudsonius) were collected in August 2005 from five sites along the Cornwall waterfront within a Hg-contaminated zone and two reference zones. The results were compared to analyses made between 1979 and 2000 by the Ontario Ministry of the Environment. Total Hg concentrations in spottail shiners from the contaminated zone were significantly higher than in reference zones, confirming previous observations. Within the contaminated zone, there were significant differences in Hg concentrations among three sites spaced about 500 m apart, consistent with a high degree of site fidelity of YOY fish and suggesting a possible internal source of Hg. Hg concentrations in spottail shiners are decreasing regionally, although year-to-year variability was high, particularly in the contaminated zone. Stable isotope analyses of spottail shiners did not reveal any differences in nitrogen isotope composition among zones that would indicate differences in food-web structure and Hg biomagnification. However, carbon sources at an upstream reference zone were not the same as within the Area of Concern. Differences in carbon isotope composition at two sites within the contaminated zone corresponded to differences in Hg concentrations, consistent with a unique internal source of Hg. The variation in Hg contamination of YOY spottail shiners over fine spatial and temporal scales provide important insights about the potential release of Hg from contaminated sediments and the role of climate in regional trends. Sessile YOY fish provide a precise indicator for demonstrating these differences and for assessing their cause. PMID- 17909882 TI - Influence of surgical staples on radiofrequency ablation using multitined expandable electrodes. AB - PURPOSE: During radiofrequency ablation (RFA), there is a risk that the multitined expandable electrode will come into contact with one of the surgical staples used to treat local recurrence after surgical operations. Our objective was to evaluate whether a surgical staple would influence the RFA of egg white using a multitined expandable electrode. METHODS: Multitined expandable electrodes, LeVeen needles (expandable diameter 3.0 cm), were sunk into an egg white bath with (a) no surgical staple, (b) a surgical staple touching one of the tines, or (c) a surgical staple touching two of the tines simultaneously. By connecting the LeVeen needle and copper plate at the bottom of the bath, RFA was then performed on the egg whites as a substitute for human tissue. Ten egg white baths were ablated under each of conditions (a), (b), and (c), for a total of 30 sets of coagulated egg white. RESULTS: There was no significant difference in the time from the power-on to the roll-off (i.e., the completion and shutting off of the electric circuit) or in the maximum diameter of the thermal lesion between conditions (a) and (b) or (a) and (c). However, the minimum diameter of the thermal lesion was significantly smaller in (c) compared with (a) (p < 0.01). CONCLUSIONS: Surgical staples have the capacity to interfere with the electromagnetic field and decrease the minimum diameter of the thermal lesion in the event that a staple touches two of the tines of a multitined expandable electrode during RFA. Although the difference might be small enough to be neglected under many clinical circumstances, we recommend that, if possible, the tines not be expanded near metallic material. PMID- 17909883 TI - Cutting balloon angioplasty (CBA) versus conventional balloon angioplasty (PTA) in the pre-dilatation of carotid artery stenosis: our preliminary experience. AB - A retrospective study was carried out to evaluate the efficacy and safety of cutting-balloon angioplasty (CBA) for the predilatation of tight fibrocalcified carotid stenosis before carotid artery stenting (CAS). We also compared the intraprocedural results of CBA with those of conventional percutaneous transluminal angioplasty (PTA) in terms of postdilatation rate. The study population was composed of 32 patients who underwent CAS requiring predilatation before stent implantation. In detail, predilatation was performed by using a conventional balloon (PTA) in 14 patients (group A) and a cutting balloon (CBA) in 18 patients (group B). Intraprocedural and periprocedural mortality, major and minor procedural and neurologic complications, arterial damage, and embolic debris found in the distal filter were recorded. We statistically compared the rate of postdilatation obtained in the two groups. Technical success was obtained in 100% of the cases. No death, major neurologic or periprocedural complications, or arterial damage were recorded in either group. Minor neurologic reactions were recorded in 1 patient from group A (4%) and in 1 patient from group B (3%). Embolic debris was found in the distal filter in 2 patients from group A (8%) and in 3 patients from group B (10%). Postdilatation was necessary in 14 patients from group A (100%) and in 9 patients from group B (50%) (p < 0.05). Although limited to a small number of patients, compared with PTA, CBA seems to be a safe and effective procedure for the predilatation of tight fibrocalcified carotid stenoses before stent placement and may decrease the need for postdilatation in this patient population. PMID- 17909884 TI - Local therapeutic results of computed tomography-guided transcatheter arterial chemoembolization for hepatocellular carcinoma: results of 265 tumors in 79 patients. AB - PURPOSE: To elucidate the local therapeutic results of computed tomography (CT) guided transcatheter arterial chemoembolization (TACE) as initial treatment for hepatocellular carcinoma (HCC), and to verify factors which affect local therapeutic results. METHODS: From 1992 to 2002, 265 tumors of 79 HCC patients were treated by 139 sessions of CT-guided TACE as initial treatment. Among these 265 tumors, 182 constituted multiple new lesions, and the remaining 83 tumors were single new lesions. Local recurrence was retrospectively ascertained on follow-up CT images obtained after TACE. RESULTS: The overall local recurrence free rates (LR-FRs) after a single TACE session at 6, 12, and 36 months were 67%, 49%, and 28%; those of the single new lesions were 80%, 66%, and 32%; and those of tumors with complete lipiodol accumulation were 82%, 68%, and 41%, respectively. LR-FRs of tumors of the single new lesions, and those of tumors with complete lipiodol accumulation, were significantly higher than the LR-FRs of multiple new lesions and tumors with incomplete lipiodol accumulation, respectively. For single new lesions < or =4 cm and the tumors that were one of multiple new lesions, there were no significant differences in the LR-FRs regarding the number of TACE sessions on the basis of patient, tumor location, or tumor size. CONCLUSION: Local therapeutic results of single new lesions were better than those of multiple new lesions, and the local therapeutic effect of TACE was not affected by the number of treatments on the basis of patient, tumor location, or tumor size. PMID- 17909885 TI - Evaluation of LightCycler as a platform for nucleic acid sequence-based amplification (NASBA) in real-time detection of enteroviruses. AB - The nucleic acid sequence-based amplification (NASBA) assay has been demonstrated to be more sensitive for detection of enteroviruses (EV) than RT-PCR. Many laboratories, however, do not have a dedicated instrument for the NASBA assay. This study aimed to evaluate the use of the Roche LightCycler as a platform for performing the NASBA assay for detection of EV. A diverse subgenera of EV were used to assess the specificity of the NASBA assay, including coxsackie, echovirus, poliovirus, and other enteroviruses together with related and unrelated viruses, including rhinovirus, respiratory syncytial virus, herpes simplex virus, adenovirus, influenza virus A, and cytomegalovirus. All species of EV tested were successfully detected using NASBA and no cross reactivity with other viruses was observed. Using serial dilutions of EV to assess sensitivity, the NASBA assay was compared to an in-house EV RT-PCR assay. The NASBA assay demonstrated a higher level of sensitivity. Fifty-one clinical samples positive for EV by viral culture were also evaluated. All NASBA results obtained were concordant with viral culture results. This study confirmed that the NASBA assay for the detection of EV could be readily performed on the LightCycler and easily incorporated into the workflow of a diagnostic laboratory equipped with a LightCycler, thereby eliminating the need for additional instrumentation. PMID- 17909886 TI - Characterization of phosphate-solubilizing fluorescent pseudomonads from the rhizosphere of seabuckthorn growing in the cold deserts of Himalayas. AB - Isolation and characterization of fluorescent pseudomonads with high phosphate solubilizing ability is reported from the alkaline and calcium-rich soils with low P availability in the cold desert region of Lahaul and Spiti in the trans Himalayas of India. Of 216 phosphate-solubilizing isolates, 12 exhibiting high solubilization of tricalcium phosphate (TCP) in NBRIP liquid culture were identified as Pseudomonas trivialis, P. poae, P. fluorescens, and Pseudomonas spp. on the basis of phenotypic features, whole-cell fatty acids methyl ester (FAME) profiles, and 16S rDNA sequencing. These isolates also showed relatively high solubilization of North Carolina rock phosphate (NCRP) in comparison to the solubilization of Mussoorie rock phosphate (MRP) and Udaipur rock phosphate (URP). The solubilization of phosphate substrates by P. trivialis and P. poae is reported for the first time. PMID- 17909887 TI - Clonal variability and its relevance in generation of new pathotypes in the spot blotch pathogen, Bipolaris sorokiniana. AB - Spot blotch pathogen Bipolaris sorokiniana of wheat was investigated with threefold objectives: to establish a relationship between morphological and pathological variability of isolates, identify clonal genotype(s) acting as a source for the generation of new variability, and to determine the mechanism of generation of such variability in the pathogen. Isolates were collected from the leaves and seeds of field-grown wheat crop at four different sites in eastern Gangetic plains of India. Eighty-six clonal isolates derived from a single isolate (gray with white patches, Group III), which segregated in an equal proportion of parental and nonparental types, were studied. Morphological characters-i.e., colony morphology, growth rate, and sporulation-were studied along with disease-causing ability of the isolate clones. Clonal isolates were grouped into three categories. Microscopic analysis of nuclei was done to determine the causes of such variability. Morphological variability appeared to be related to the pathological variability. The isolate having epidemic potential appeared different than that acting as the reservoir for variability. The cause of such variability could be attributed either to hyphal fusion and heterokaryosis, nuclear migration and occurrence of multinucleate state, or a combination of these factors. Random Amplified Polymorphic DNA (RAPD) assay suggested that the unique fragments for different groups could be utilized as molecular markers to identify the isolates of specific groups. PMID- 17909888 TI - Conjugal transfer of plasmid R6K gamma ori minireplicon derivatives from Escherichia coli to various genera of pathogenic bacteria. AB - Three R6K-derived gamma ori minireplicons were successfully transferred by conjugation from Escherichia coli to several species of pathogenic bacteria. The pFL129 replicon encodes the wild-type initiation replication protein pi, while plasmids pFL130 and pAG101 encode mutant forms of the pi protein conferring the plasmid copy-up phenotype. Plasmids could be transferred to all recipient species tested, although high efficiency conjugal transfer was only obtained with genera of the Enterobacteriaceae. The efficiency of plasmid transfer to all recipients was lower for the copy-up derivatives, pFL130 and pAG101, than for pFL129. The three gamma ori replicons were stably maintained in all transconjugants except pFL129 in Listeria monocytogenes. The two mutant plasmids retained their copy-up phenotype in the new bacterial hosts. PMID- 17909889 TI - Expression and regulation of the yggG gene of Escherichia coli. AB - Our previous study indicated that Era, a membrane-associated GTPase essential for the survival of Escherichia coli, binds with the product of the yggG gene. However, the expression, regulation, and function of the yggG gene have not been established. In this study, the transcript of the yggG gene was determined by analysis of the 5'-end of the yggG mRNA using 5'-RACE (5'-rapid amplification of cDNA ends) method. The promoter and transcription regulatory regions of yggG were analyzed through systematic analysis of the transcriptional activity of the fragment containing a 339-bp 5' flanking sequence of yggG mRNA. The results showed that the sequence -39/-1 upstream of the transcriptional start site of the yggG gene contained a core promoter required for the expression of 25-kDa YggG protein, whereas the -106/-40 region was associated with transcriptional upregulation of yggG under heat shock. Immunocytochemistry and subcellular fraction analysis showed that YggG was a membrane-associated protein. Based on these results, we confirmed that the -39/780 region contains the whole set of the promoter and coding sequence of the yggG gene. The expression regulation of the yggG gene under stress conditions and the YggG protein located on cell membrane are consistent with the bioinformatics analysis that YggG, a metallopeptidase, is functional as a heat shock protein that is associated with Era functions. PMID- 17909890 TI - Expression, purification, and characterization of a [Fe2S2] cluster containing ferredoxin from Acidithiobacillus ferrooxidans. AB - The [2Fe-2S] cluster containing ferredoxin has attracted much attention in recent years. Genetic analyses show that it has an essential role in the maturation of various iron-sulfur (Fe-S) proteins and functions as a component of the complex machinery responsible for the biogenesis of Fe-S clusters. The gene of ferredoxin from A. ferrooxidans ATCC 23270 was cloned, successfully expressed in Escherichia coli, and purified by one-step affinity chromatography to homogeneity. The MALDI TOF MS and spectra results of the recombinant protein confirmed that the iron sulfur cluster was correctly inserted into the active site of the protein. Site directed mutagenesis results revealed that Cys42, Cys48, Cys51, and Cys87 were ligating with the [Fe(2)S(2)] cluster of the protein. PMID- 17909891 TI - A deletion mutation of the protein tyrosine phosphatase kappa (Ptprk) gene is responsible for T-helper immunodeficiency (thid) in the LEC rat. AB - Bone marrow (BM)-derived T-cell progenitors differentiate into CD4 or CD8 single positive (SP) cells in the thymus. We have previously reported that a single autosomal mutation, thid, causes a defect in the maturation of CD4 SP thymocytes and an abnormality of peripheral helper T cells in the LEC rat. In this study we attempted to identify a gene responsible for the thid mutation. We first performed genetic linkage analysis and mapped the thid locus between Myb and D1Rat392 on Chr 1. In this region we found an approximately 380-kb deletion from intron 3 of the Ptprk gene, which encodes a receptor-like protein tyrosine phosphatase type kappa (RPTPkappa) to intron 1 of the RGD1560849 predicted gene in the LEC rat genome. Reconstitution with syngenic BM cells transduced Ptprk but not the RGD1560849 predicted gene rescued development of CD4 SP cells in the LEC rat thymus. It is confirmed by this result that the Ptprk gene is responsible for the thid mutation in the LEC rat. Our results further suggest that RPTPkappa plays a critical role in the development of CD4 SP cells in the thymus. PMID- 17909892 TI - Is reflux cough due to gastroesophageal reflux disease or laryngopharyngeal reflux? AB - Reflux is a common cause of chronic cough. Surveys of patients with chronic cough point to a high association with gastroesophageal disease. Because of our bipedalism and speech, humans are prone to both reflux and aspiration. Whether the reflux of stomach contents into the esophagus alone is sufficient to cause cough or whether reflux into the upper airway is required is unknown. In 50 consecutive patients with chronic cough, symptoms of laryngopharangeal reflux (LPR) paralleled those of gastroesophageal reflux, suggesting no unique syndrome of LPR but that it is part of the protean manifestations of reflux disease. PMID- 17909893 TI - Spontaneous pneumomediastinum may be associated with both anorexia nervosa and obesity. PMID- 17909894 TI - Before we get started: what is a cough? AB - Cough is an airway defensive reflex consisting of an inspiratory phase followed by a forced expiratory effort initially against a closed glottis, followed by active glottal opening and rapid expiratory flow. The expiration reflex (ER) differentiates from cough for the lack of a preparatory inspiration. The reflexes subserve different functions: cough will clear the lower airways from debris and mucus, while the expiration reflex will prevent aspiration. Clinically, a cough epoch is a sequence of motor acts resulting from a combination of true coughs and ERs that need to be accurately identified and measured for adequate quantitative description. PMID- 17909895 TI - The peroxisome proliferator activated receptor gamma (PPARgamma) ligand rosiglitazone modulates bronchoalveolar lavage levels of leptin, adiponectin, and inflammatory cytokines in lean and obese mice. AB - Obese mice that lack leptin receptor (db (-) /db (-)) have been shown to have innate bronchial hyperresponsiveness (BHR). It has been proposed that the obesity mediated BHR may involve a combination of increased leptin and reduced systemic adiponectin levels. The aim of this study was to determine if obesity modifies the airway concentration of leptin and adiponectin and whether treatment with a synthetic peroxisome proliferator-activated receptor gamma (PPARgamma) ligand can reduce airway leptin and increase airway adiponectin. In this study, obese, leptin receptor-deficient (db (-) /db (-)), or lean (db ( + ) /db (-)) mice were treated with rosiglitazone (3 mg/kg/day) or vehicle by gavage daily for 1 week. Bronchioalveolar lavage (BAL) was subsequently performed to determine levels of leptin, adiponectin, and inflammatory cytokines. Treatment with rosiglitazone increased BAL adiponectin levels in lean (p = 0.04) and to a lesser extent in obese mice (p = 0.07). Rosiglitazone treatment lowered leptin levels in lean mice, but increased leptin levels in BAL fluid of obese mice (p < 0.01). The BAL levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) were lower in the lean rosiglitazone-treated group compared with the obese vehicle-treated group and lower in the obese rosiglitazone-treated group compared with the obese vehicle-treated group. These results demonstrate that obesity is associated with alterations in adipokine and cytokine levels in the airways that can be modulated by treatment with roziglitazone. PMID- 17909896 TI - Older-generation antihistamines and cough due to upper airway cough syndrome (UACS): efficacy and mechanism. AB - Older-generation H1 antihistamines are recommended in empiric protocols for the treatment of cough due to upper airway cough syndrome (UACS). Data from double blind placebo-controlled trials that support the use of older-generation antihistamines in UACS are limited to a single study of cough due to the common cold. However, several empiric trials strongly support efficacy of older generation antihistamines in patients with chronic cough. Data from a variety of studies support the concept that newer-generation H1 antihistamines are not useful in the treatment of cough due to UACS. The mechanism of action of older generation H1 antihistamines has been proposed to be anticholinergic activity, but the rank order potency of these drugs as muscarinic receptor antagonists is not consistent with this hypothesis. Actions of these drugs on histamine H1 and/or nonhistaminergic receptors in the central nervous system remain a possible explanation for their effects on cough due to UACS. The effects of older H1 antihistamines may also be molecule specific, rather than attributable to the entire class of compounds. Additional studies should be performed to document the activity of these drugs in double-blind placebo-controlled trials. PMID- 17909898 TI - The First American Cough Conference. New York City, June 8-9, 2007. PMID- 17909897 TI - Currently available cough suppressants for chronic cough. AB - Chronic cough is a common symptom but only a fraction of patients seek medical attention. Addressing the causes of chronic cough may lead to control of cough; however, this approach is not always successful since there is a certain degree of failure even when the cause(s) of cough are adequately treated; in idiopathic cough, there is no cause to treat. Persistent cough may be associated with deterioration of quality of life, and treatment with cough suppressants is indicated. Currently available cough suppressants include the centrally acting opioids such as morphine, codeine, and dextromethorphan. Peripherally acting antitussives include moguisteine and levodropropizine. Early studies report success in reducing cough in patients with chronic bronchitis or COPD; however, a carefully conducted study showed no effect of codeine on cough of COPD. Success with these cough suppressants can be achieved at high doses that are associated with side effects. Slow-release morphine has been reported to be useful in controlling intractable cough with good tolerance to constipation and drowsiness. There have been case reports of the success of centrally acting drugs such as amitryptiline, paroxetine, gabapentin, and carbamezepine in chronic cough. New opioids such as nociceptin or antagonists of TRPV1 may turn out to be more effective. Efficacy of cough suppressants must be tested in double-blind randomised trials using validated measures of cough in patients with chronic cough not responding to specific treatments. Patients with chronic cough are in desperate need of effective antitussives that can be used either on demand or on a long-term basis. PMID- 17909900 TI - Gastric bypass: increased restriction for poor weight loss. PMID- 17909899 TI - Preclinical assessment of novel therapeutics on the cough reflex: cannabinoid agonists as potential antitussives. AB - Cough, a reflex defense mechanism, is a common symptom of many airway inflammatory diseases. At present there are no satisfactory treatments for cough that have an acceptable side effect profile. Recent data have described the inhibitory effect of selective cannabinoid CB(2) receptor agonists on sensory nerve activity in vitro and the cough reflex in a guinea pig model. CB(2) receptor expression is limited in the central nervous system (CNS) and hence the development of selective agonists may provide a new therapeutic strategy for treatment of cough devoid of the CNS-mediated side effects that are normally associated with nonselective cannabinoid agonists. PMID- 17909902 TI - St. Mark's incontinence score. PMID- 17909901 TI - Phylogenetic relationships of South China Sea snappers (genus Lutjanus; family Lutjanidae) based on mitochondrial DNA sequences. AB - Phylogenetic relationships of intra- and interspecies were elucidated based on complete cytochrome b (cyt b) and cytochrome c oxidase subunit II (COII) gene sequences from 12 recognized species of genus Lutjanus Bloch in the South China Sea (SCS). Using the combined data set of consensus cyt b and COII gene sequences, interspecific relationships for all 12 recognized species in SCS were consistent with Allen's morphology-based identifications, with strong correlation between the molecular and morphological characteristics. Monophyly of eight species (L. malabaricus, L. russellii, L. stellatus, L. bohar, L. johnii, L. sebae, L. fulvus, and L. fulviflamma) was strongly supported; however, the pairs L. vitta/L. ophuysenii and L. erythropterus/L. argentimaculatus were more similar than expected We inferred that L. malabaricus exists in SCS, and the introgression caused by hybridization is the reason for the unexpectedly high homogeneity. PMID- 17909903 TI - A novel concept for the surgical anatomy of the perineal body. AB - PURPOSE: Perineal body is considered by investigators as a fibromuscular structure that is the site of insertion of perineal muscles. We investigated the hypothesis that perineal body is the site across which perineal muscles pass uninterrupted from one side to the other. METHODS: Perineal body was studied in 56 cadaveric specimens (46 adults, 10 neonatal deaths) by direct dissection with the help of magnifying loupe, fine surgical instruments, and bright light. RESULTS: Perineal body consisted of three layers: 1) superficial layer, which consisted of fleshy fibers of the external anal sphincter extending across perineal body to become the bulbospongiosus muscle; 2) tendinous extension of superficial transverse perineal muscle crossing perineal body to contralateral superficial transverse perineal muscle, with which it formed a criss-cross pattern; and 3) tendinous fibers of the deep transverse perineal muscle; the fibers crossing perineal body decussated in criss-cross pattern with the contralateral deep transverse perineal muscle. A relation of levator ani or puborectalis muscles to perineal body could not be identified. CONCLUSIONS: Perineal body (central perineal tendon) is not the site of insertion of perineal muscles but the site along which muscle fibers of these muscles and the external anal sphincter pass uninterrupted from one side to the other. Such a free passage from one muscle to the other seems to denote a "digastric pattern" for the perineal muscles. Perineal body is subjected to injury or continuous intra abdominal pressure variations, which may eventually result in perineocele, enterocele, or sigmoidocele. PMID- 17909904 TI - Activin a causes cancer cell aggressiveness in esophageal squamous cell carcinoma cells. AB - BACKGROUND: Expression of activin A is associated with lymph node metastasis and clinical stage in esophageal cancer. METHODS: To clarify the aggressive behavior of tumors with high activin A expression, we used the beta subunit of activin A to establish stable activin betaA (Act-betaA)-transfected carcinoma cells in two human esophageal carcinoma cell lines, KYSE110 and KYSE140. The biological behavior of these cells was compared with that in mock-transfected cells from the same cell lines. We focused our attention on cell growth and tumorigenesis, and proliferation and apoptosis. RESULTS: Both Act-betaA-transfected carcinoma cell lines showed a higher growth rate than the mock-transfected carcinoma cells. In an in vitro invasion assay and a xenograft analysis, the Act-betaA-transfected carcinoma cells showed far higher proliferation in vitro and a higher potency for tumorigenesis in vivo, respectively. Moreover, in an analysis of apoptosis via Fas stimulation, the Act-betaA-transfected carcinoma cells showed a higher tolerance to apoptosis compared with the mock-transfected carcinoma cells. Moreover, anti-activin-neutralizing antibody-treated squamous cell cancer cell lines inhibited their migration. CONCLUSIONS: Collectively, these data indicate that continuous high expression of activin A in esophageal carcinoma cells is not related to tumor suppression, but rather to tumor progression in vitro and in vivo. The inhibition of activin might be one of the methods to attenuate tumor aggressiveness. PMID- 17909905 TI - Epithelioid sarcoma: prognostic factors and survival in a series of patients treated at a single institution. AB - BACKGROUND: Epithelioid sarcoma (ES) is a rare subtype of soft-tissue sarcoma of unknown histogenesis. Typically, it occurs superficially as single/multiple nodules (nodular ES), or in deeper tissues as a mass. The correlation between initial presentation and clinical outcome was investigated. METHODS: Fifty-four consecutive patients surgically treated at a single referral center were retrospectively reviewed. Thirty-six patients presented with a primary and 18 with a recurrent tumor. Potential prognostic clinicopathological variables, including macroscopic features at first presentation, were tested by univariable and multivariable analysis with respect to overall (OS), metastasis-free (MFS), and local recurrence-free survival (LRFS). RESULTS: The 10-year OS was 61.8% for the whole series. Thirty patients relapsed; in detail, local and distant failure occurred in 14 (25.9%) and 24 (44.4%) patients, respectively. The lymph node involvement rate was 16/54 (29.6%). In both the whole series and the subset of patient with primary ES, single localized tumor correlated with increased OS at multivariable analysis; occurrence of nodal involvement during postoperative follow-up correlated to worse OS and MFS. Nodular ES was an independent predictor of worse LRFS. In univariable analysis, nodular ES was associated with smaller tumor size, distal limb locations, earlier classification of malignant tumor (TNM) stage, and higher amputation rate. A statistical difference in the pattern of failure between nodular and mass ES was found. CONCLUSIONS: Primary tumor macroscopic features seem to correlate to different local aggressiveness and failure patterns. Better prognosis is associated with single localized disease stage and no occurrence of locoregional spread. PMID- 17909906 TI - Dosimetric evaluation of radiation exposure during I-125 vicryl mesh implants: implications for ACOSOG z4032. AB - BACKGROUND: Segmentectomy or wedge resection along with brachytherapy delivered via a vicryl mesh implant imbedded with 125I is a novel therapeutic modality to treat early stage lung cancer. This modality is being evaluated in a large national prospective randomized trial (ACOSOG Z4032). There has been concern that this method exposes physicians and staff to unacceptable amounts of radiation. In this prospective study, we measured the exposure to health care professionals during such a procedure. METHODS: Dosimetric readings using Special Microdosimeter thermoluminescent detectors (TLDs) (Landauer, Inc) were performed during 22 125I vicryl mesh implantations. Diodes were placed on the back of the each hand of the primary radiation oncologist and primary surgeon during the creation and implantation of the mesh. In addition, diodes were placed on the posterior shoulder of the patient to obtain a control reading. RESULTS: Patients had 40-60 125I seeds placed. Median activity per seed was 0.511 milli Curie (mCi), with a median total activity implanted of 23.0 mCi. Median radiation dose to the radiation oncologist was 1 milli rem (mrem), and that to the surgeon was 2 mrem. Median dose to the control diode on the patient was a median radiation dose to the outside of the patient of 5.4 mrem/h. CONCLUSIONS: There is very little radiation exposure to physicians and staff during a segmentectomy and 125I vicryl mesh implantation. This is a safe method of lung cancer treatment with respect to health care professionals, although the ALARA (As Low As Reasonably Achievable) principle should still be followed. PMID- 17909907 TI - Lymphadenectomy in surgical oncology. PMID- 17909908 TI - Practice pearl: a novel use of tranexamic Acid for decreasing the blood loss of aneurysmal bone cyst ablation. PMID- 17909909 TI - Noninvasive pancreatic cystic neoplasms can be safely and effectively treated by limited pancreatectomy. AB - BACKGROUND: Appropriate management of cystic lesions of the pancreas is controversial. Major pancreatectomies (pancreaticoduodenectomy or distal pancreatectomy with splenectomy) are the commonly used procedures, even though most cystic lesions are noninvasive neoplasms. We tested the adequacy of limited pancreatectomies in the treatment of pancreatic cystic lesions. METHODS: Data from 109 patients who underwent surgical resection of a pancreatic cystic lesion at National Taiwan University Hospital from 2001 to 2007 were retrospectively reviewed. Major pancreatomies (n = 79) constituted pancreaticoduodenectomy and total/distal pancreatectomies, while other resection procedures (n = 30) represented limited pancreatectomies. Clinicopathologic features were compared between the major and limited groups. RESULTS: There were no statistically significant differences in sex, age, presence of symptoms, cyst diameter, minor or major treatment complications, or pancreatic leakage between the two groups. Cystic lesions located in the neck/body/tail rather than in the head/uncinate process were significantly more often treated with limited pancreatectomy (P = .02). Both groups had similar pathologic distribution of cystic lesions, with the exception of nine invasive neoplasms. The latter were treated with major pancreatectomy. No recurrence was noted in 100 patients with noninvasive cystic neoplasms after major or limited pancreatectomy. CONCLUSIONS: Noninvasive pancreatic cystic neoplasms can be safely and effectively treated by limited pancreatectomy. PMID- 17909911 TI - Improved long-term outcome of surgery for advanced colorectal liver metastases: reasons and implications for management on the basis of a severity score. AB - BACKGROUND: The outcome of liver resection for colorectal liver metastases (CRLM) appears to be improving despite the fact that surgery is offered to patients with more-severe disease. To quantify this assumption and to understand its causes we analyzed a series of patients on the basis of a standardized severity score and changes in management occurring over the years. METHODS: Patients' characteristics, operative data, chemotherapies and follow-up were recorded. CRLM severity was quantified according to Fong's clinical risk score (CRS), modified to take into account the presence of bilateral liver metastases. Three periods were analyzed, in which different indications, surgical strategies and uses of chemotherapy were applied: 1984-1992, 1993-1998, and 1999-2005. RESULTS: Between January 1984 and December 2005, 210 liver resections were performed in 180 patients (1984-1992, 43 patients; 1993-1998, 42 patients; 1999-2005, 95 patients). CRLM severity increased throughout the time periods, as did the use of neoadjuvant chemotherapies, repeat resections, and multistep procedures. While the disease-free survival did not improve over time, the 1-, 3- and 5-year overall survival rate increased from 85%, 30%, and 23% in the first period, to 88%, 60%, and 34% in the second period, and to 94%, 69%, and 46% in the third period. CONCLUSIONS: Analysis according to the CRS showed that despite the fact that patients had more severe disease, the overall survival improved over the years, mainly thanks to more aggressive treatment of recurrent disease. Management of advanced CRLM should, from the start, take into account the likelihood of secondary procedures. PMID- 17909912 TI - Treatment guidelines for branch duct type intraductal papillary mucinous neoplasms of the pancreas: when can we operate or observe? AB - BACKGROUND: The objectives of this study were to investigate the clinicopathological features of branch intraductal papillary mucinous neoplasm (IPMN) and to determine safe criteria for its observation. Most clinicians agree that surgical resection is required to treat main duct-type IPMN because of its high malignancy rate. However, no definite treatment guideline (with respect to surgery or observation) has been issued on the management of branch duct type IPMN. METHODS: We retrospectively reviewed the clinicopathological data of 138 patients who underwent operations for IPMN between 1993 and 2006 at five institutes in Korea. RESULTS: Of 138 patients (mean age, 60.6 years; 87 men, 51 women), 76 underwent pancreatoduodenectomy, 39 distal pancreatectomy, 4 total pancreatectomy, and 20 limited pancreatic resection. There were 112 benign cases: 47 adenoma, 63 borderline cases, and 26 malignant cases, with 9 of these being noninvasive and 17 invasive. By univariate analysis, tumor size and the presence of a mural nodule were identified as meaningful predictors of malignancy. By receiver operating characteristic curve analysis, a tumor size of >2 cm was found to be the most valuable predictor of malignancy. When cases were classified according to tumor size and the presence of a mural nodule, the malignancy rate for a tumor 2 cm, >25%. CONCLUSIONS: Many branch duct IPMNs are malignant. Surgical treatment is recommended, except in cases that are strongly suspected to be benign or cases that present a high operative risk. Observation is only recommended in patients with a tumor size of 50% pathologic response to treatment. Median follow-up after resection for all patients was 25 months. Median DSS for all patients was 33 months. Three-year DSS improved from 44% to 69% with at least a 50% histologic response (P = .01). Factors associated with decreased DSS included positive nodes at resection, pT3 tumor or greater, high grade, perineural or vascular invasion, and <50% response. Multivariate analysis identified nodal status and perineural or vascular invasion as independent predictors of survival. CONCLUSIONS: Posttreatment nodal status and perineural or vascular invasion at resection, but not graded histologic response, independently predict DSS after neoadjuvant chemotherapy and surgical resection of gastric cancer. PMID- 17909918 TI - Incidence of postoperative pancreatic fistula and hyperamylasemia after cytoreductive surgery and hyperthermic intraperitoneal chemotherapy. AB - INTRODUCTION: The purpose of this study was to analyze the postoperative pancreatic morbidity of cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) in the treatment of peritoneal surface malignancies (PSM). PATIENTS AND METHODS: Two hundred and sixty five patients (87M/178F) with PSM underwent 270 consecutive procedures. The mean age was 52 years (range: 22-79 years). CRS was performed using peritonectomy procedures. HIPEC through the closed abdomen technique was conducted using cisplatin (CDDP 25 mg/m2/L of perfusate)+mitomycin C (MMC 3.3 mg/m2/L of perfusate) or CDDP (43 mg/L of perfusate)+doxorubicin (Dx 15.25 mg/L of perfusate), at 42.5 degrees C. Diagnosis and classification of postoperative pancreatic fistula (POPF) were performed according to the international study group on pancreatic fistula criteria. Serum amylase alterations were graded according to the National Cancer Institute (NCI) common terminology criteria for adverse events (CTCAE) v3. RESULTS: POPF was observed in 13 (4.8%) cases. Three cases were classified as major (grade C). Two cases presented postoperative pancreatitis. G3-4 alteration of amylase was observed in 12.3% of the cases. Performing splenectomy and CDDP dosage for HIPEC >240 mg were proven to be independent risk factors for both G3-4 hyperamylasemia and POPF. CONCLUSIONS: CRS+HIPEC presented an acceptable rate of pancreatic morbidity which did not contribute to the mortality related to the procedure. Most of the POPF were mild and/or easily controlled by conservative measures. Although not specific a normal amylasemia could be a useful marker of pancreatic integrity after CRS+HIPEC. PMID- 17909919 TI - Optimal control of gypsy moth populations. AB - This study investigates an optimal strategy for the cost effective control of gypsy moth populations. Gypsy moth populations cycle between low sparse numbers to high outbreak levels and it is during the outbreak levels that the moths cause extensive damage to plant foliage which can lead to deforestation. Deforestation can result in significant economic damage to infested areas, and consequently, there have been many efforts to control moth populations. One effective method of control is the use of the biocontrol agent, Gypchek, but its production is costly. We develop a mathematical model which combines population dynamics and optimal control of the moth population to explore strategies by which the total cost of the gypsy moth problem (economic damage and cost of Gypchek) can be minimized. PMID- 17909920 TI - Optimal reproduction strategies in two species of mound-building termites. AB - We formulate a mathematical model for food collection and production of workers and nymphs in 2 species of mound building termites. We maximise the number of nymphs (reproductives) produced by each colony over its lifetime with respect to the proportion of eggs that hatch as nymphs as opposed to workers. The results predict that food storage has a very important influence on the pattern of nymph and worker production. Food storage affects the part of the year that nymph production dominates, whether nymphs and workers are produced at the same time or not, and the existence of a final phase in the colony's life when a very large number of nymphs but no workers are produced. PMID- 17909921 TI - Esophagectomy for high grade dysplasia is safe, curative, and results in good alimentary outcome. AB - BACKGROUND: The increasing adoption of endoscopic therapies and expectant surveillance for patients with high grade dysplasia (HGD) in Barrett's esophagus has created considerable controversy regarding the ideal treatment choice. Confusion may be due, in part, to a limited understanding of the outcomes associated with surgical resection for HGD and extrapolation of data derived from patients undergoing an esophagectomy for invasive cancer. The purpose of our study was to document the perioperative and symptomatic outcomes and long-term survival after esophagectomy for HGD of the esophagus. MATERIAL AND METHODS: The study population consisted of 38 patients who underwent esophagectomy for biopsy proven HGD between 10/1999 and 6/2005. Three patients were excluded from analysis due to obvious tumor on upper endoscopy. Patients were evaluated regarding ten different foregut symptoms and administered a ten-question appraisal of eating and bowel habits. Outcome measures included postoperative morbidity and mortality, the prevalence of invasive cancer in the esophagectomy specimens, symptomatic and functional alimentary results, patient satisfaction, and long term survival. Median follow-up was 32 months (range, 7-83). RESULTS: Thirty-day postoperative and in-hospital mortality was zero. Complications occurred in 37% (13/35), and median length of stay was 10 days. Occult adenocarcinoma was found in 29% (10/35) of surgical specimens (intramucosal in four; submucosal in five; and intramuscular in one with a single positive lymph node.) Patients consumed a median of three meals per day, most (76%, 26/34) had no dietary restrictions, and two-thirds (23/34) considered their eating pattern to be normal or only mildly impacted. Meal size, however, was reported to be smaller in the majority (79%, 27/34) of patients. Median body mass index (BMI) decreased slightly after surgery (28.6 vs 26.6, p>0.05), but no patient's BMI went below normal. The number of bowel movements/day was unchanged or less in a majority (82%) of patients after surgery. Fifteen of 34 (44%) patients reported loose bowel movements, which occurred less often than once per week in 10 of the 15. One patient had symptoms of dumping. Mean symptom severity scores improved for all symptoms except dysphagia and choking. Four patients developed foregut symptoms that occurred daily. Most patients (82%) required at least one postoperative dilation for dysphagia. Almost all (97%) patients were satisfied. Disease-free survival was 100%, and overall survival was 97% (34/35) at a median of 32 months. CONCLUSION: Esophagectomy is an effective and curative treatment for HGD and can be performed with no mortality, acceptable morbidity, and good alimentary outcome. These data provide a gold standard for comparison to alternative therapies. PMID- 17909922 TI - Acute pancreatitis and pregnancy: a 10-year single center experience. AB - BACKGROUND: Acute pancreatitis in pregnancy is rare. We report our institutional therapeutic approaches to this disease and its effect on maternal and fetal outcomes. METHODS: A retrospective review of medical records of pregnant women admitted to Brigham and Women's Hospital between 1996 and 2006. RESULTS: Twenty one patients, presenting with 34 episodes of acute pancreatitis were identified. Most attacks (56%) occurred in the second trimester. Twelve patients had biliary pancreatitis. Three had pancreatitis secondary to other causes and six had "undetermined" etiologies. Of those with biliary pancreatitis, six underwent cholecystectomy; in a third of these cases, initial conservative therapy had failed. The other six patients underwent endoscopic sphincterotomy (n=2) or conservative therapy (n=4). Fifty percent of the patients with biliary pancreatitis managed conservatively had a recurrent episode of pancreatitis vs none in the cholecystectomy group. There was no significant difference in length of hospital stay between the three treatment groups (cholecystectomy, sphincterotomy, and conservative therapy). No maternal deaths were observed; there were four preterm labors and one fetal loss. CONCLUSION: If treated conservatively, pregnant patients with biliary pancreatitis appear to have a high recurrence rate. Early surgical intervention is appropriate, safe, and does not increase the length of hospital stay. PMID- 17909924 TI - Stapled hemorrhoidectomy versus conventional excision hemorrhoidectomy for acute hemorrhoidal crisis. AB - We compared the safety and clinical outcomes of stapled hemorrhoidectomy and conventional excision hemorrhoidectomy in the treatment of acute hemorrhoidal crisis, and analyzed various factors associated with complications in stapled hemorrhoidectomy. Forty patients underwent stapled hemorrhoidectomy and forty underwent conventional excision hemorrhoidectomy. All had the operation under local anesthesia with conscious sedation within 24 h of admission. The length of surgery, hospital stay, disability, postoperative pain, and the use of analgesics were significantly less for patients in the stapled hemorrhoidectomy group. Stapled hemorrhoidectomy did not significantly increase the rate of complications. Five patients in the stapled group (12.5%) required further surgical intervention: three with thrombosed hemorrhoids and two with recurrent prolapse. No serious complications were reported in either group. Patient satisfaction was similar in the two groups. Increased age was identified as a factor that significantly elevated the risk of complications in the stapled group (OR, 1.06; 95% CI, 1.01-1.13). Anemia and time between the onset of prolapsed hemorrhoids and hospital admission were also risk factors for complications, although they were not significant. Stapled hemorrhoidectomy is a feasible treatment for selected patients with an acute hemorrhoidal crisis and has a similar complication rate to that of conventional excision hemorrhoidectomy. Stapled hemorrhoidectomy is superior in less-postoperative pain, shorter operation time, shorter hospital stay, and earlier return to normal activity. However, we suggest that older patients with anemia or a prolonged hemorrhoidal crisis are unsuitable for stapled hemorrhoidectomy. PMID- 17909923 TI - Laparoscopic vs. open resection of noninvasive intraductal pancreatic mucinous neoplasms. AB - Required resection margins for noninvasive intraductal papillary mucinous neoplasms (IPMNs) are a controversial issue. Over a 10-year period we have resected IPMNs from the entire pancreatic gland with minimally invasive techniques and compared our survival and complication rates with open controls to see if any difference in resection margins and outcomes could be observed. Data were collected retrospectively, including our first cases of advanced laparoscopic resections. Five-year Kaplan-Meier curves were calculated and statistical analysis was performed using the log rank and Student's T test for continuous variables. Chi square and Fisher's exact tests were used for analyzing categorical variables. From March 1997 to February 2006, we operated on 22 patients with noninvasive IPMNs, of which 9 (41%) were operated on laparoscopically and 13 (59%) using open techniques. Three patients underwent laparoscopic duodenopancreatectomy, compared to five in the open group. All resection margins were negative, but two patients required total pancreatectomy, both of which were performed laparoscopically. One of these was converted to open (11%) because of difficulty in reconstructing the biliary anastomosis. The overall complication rates were 56% for the laparoscopic group and 85% for the open group. Twenty-two percent of the laparoscopic group required reoperation and 11% required percutaneous drainage, compared to 15 and 23% in the open group, respectively. All patients are alive after a mean of 20 months (range = 2-43) in the laparoscopic group and 37 months (range = 1-121) in the open one (p > 0.05). Laparoscopic resection of noninvasive IPMNs of the entire pancreatic gland has similar complication and survival rates as open procedures. As a result, the laparoscopic approach is appropriate for noninvasive IPMNs of the entire pancreatic gland; however, larger cohorts are needed to see if any approach has superior outcomes. Because of these favorable results, studies are currently underway to see if the minimally invasive approach is also appropriate for invasive IPMNs. PMID- 17909925 TI - Secondary chondrosarcoma in cartilage bone tumors: report of 32 patients. AB - BACKGROUND: Secondary malignancies arising from benign bone tumors are rare. Their recognition and diagnosis are difficult, and their slow growth and late recurrence require long-term follow-up. In this study, malignant transformation rates of various histological types of benign cartilage-forming bone tumors in large series were evaluated. METHODS: Between 1986 and 2004, a retrospective analysis of 627 cartilage-forming benign bone tumors revealed that 32 patients had malignant transformation. Of the 32 patients, 14 had solitary osteochondromas, 10 had multiple osteochondromas, 6 had a solitary enchondroma, 1 had Ollier's disease, and 1 had Maffucci's syndrome. The patient with Ollier's disease had two chondrosarcomas, and one patient with multiple osteochondroma had three chondrosarcomas. The cases were included in the study only when complete clinical documentation, radiological records, and histological analyses were available. RESULTS: The rate of malignant transformation for cartilage originating tumors was 5.1% (solitary osteochondromas 4.2%, multiple osteochondromas 9.2%, solitary enchondromas 4.2%). The average time between the initial diagnosis and malignant transformation was 9.8 years. The most common site of involvement was the proximal portion of the femur. The tumors generally were well differentiated. The mean follow-up period was 57.3 months. Five patients (15.6%) died of tumor recurrence or metastasis at an average of 20.6 months. One patient is alive with tumor at 104 months. CONCLUSIONS: Cartilage forming benign bone tumors are rather prone to undergo malignant transformation. Although malignant transformation of a benign bone tumor is a rarely encountered situation, orthopedic surgeons should be cautious while following patients with a benign bone neoplasm. Early recognition and appropriate surgical treatment are required to achieve successful outcomes. The rate of local recurrence in secondary chondrosarcomas depends not only on adequate surgical treatment but also on the localization and histological grade. PMID- 17909926 TI - Extraskeletal osteosarcoma in Japan: multiinstitutional study of 20 patients from the Japanese Musculoskeletal Oncology Group. AB - BACKGROUND: The clinical features of extraskeletal osteosarcoma are not well known. Unlike osteosarcoma of the bone, the efficacy of chemotherapy for extraskeletal osteosarcoma has not been established yet. METHODS: A multiinstitutional study of extraskeletal osteosarcoma was conducted. RESULTS: A total of 20 patients were enrolled in the study from the Japanese Musculoskeletal Oncology Group institutions. Their ages ranged from 15 to 88 years (mean 50 years). Six lesions were located in the thigh, four in the chest, four in the buttocks, three in the upper arm, and others. UICC surgical stages were II in 5 patients, III in 13, and IV in 2. Surgical resection was performed in 19 patients, and the consequent surgical margin was wide in 15 patients, marginal in 3, and intralesional in 1. The overall 5-year survival rate was 66%. Three patients demonstrated tumor recurrence postoperatively, and the 5-year local control survival rate was 75%. Altogether, 15 patients received chemotherapy, of whom 11 had evaluable responses as follows: complete response in none, partial response in 5 patients, and no change or progressive disease in 6 patients, with a response rate of 45%. CONCLUSIONS: Among the patients with extraskeletal osteosarcoma, both the 5-year survival rate and the chemotherapy response rate tended to improve in this study in comparison to the findings published in previous reports. As a result, we believe that treatment regimens that include systemic chemotherapy may be able to improve the prognosis in patients with extraskeletal osteosarcoma. PMID- 17909927 TI - Cross-sectional anatomy in postdistraction osteogenesis tibia. AB - BACKGROUND: Although the early period of distraction osteogenesis has been extensively investigated, there are few data describing the appearance of medium term bone regeneration. METHOD: We investigated 10 adults with magnetic resonance imaging scans. Seven of them underwent bone transport, and three had tibial lengthening. The mean follow-up was 28 months after removal of the external fixator. The values were compared with those of the contralateral tibia, which acted as a control. RESULTS: All of the cases with bone transport had an increase in the volume of the whole tibia of 15.3%-50.8%. The diameters of the regenerated segments increased significantly (P < 0.0001) in all cases. The mean signal intensity in the regenerate decreased significantly in seven cases (P < 0.0001), which suggested a rise in the content of unhydrated tissue, such as bone and collagen. The cross-sectional area of the transported segment increased in all cases (P < 0.01). Finally, in the patients who underwent bone transport, the docking site was seen to be obstructed by unhydrated tissue. CONCLUSIONS: Contrary to previous claims, the postdistraction osteogenesis of tibia consists of areas with potentially different biomechanical properties. Recognition of these changes is essential not only for appropriate preoperative counseling but also for considering treatment modalities in case of fracture. PMID- 17909928 TI - Interrater reliability of identifying indicators of posterior ligamentous complex disruption when plain films are indeterminate in thoracolumbar injuries. AB - BACKGROUND: The Spine Trauma Study Group (STSG) has proposed a novel thoracolumbar injury classification system and score (TLICS) in an attempt to define traumatic spinal injuries and direct appropriate management schemes objectively. The TLICS assigns specific point values based on three variables to generate a final severity score that guides potential treatment options. Within this algorithm, significant emphasis has been placed on posterior ligamentous complex (PLC) integrity. The purpose of this study was to determine the interrater reliability of indicators surgeons use when assessing PLC disruption on imaging studies, including computed tomography (CT) and magnetic resonance imaging (MRI). METHODS: Orthopedic surgeons and neurosurgeons retrospectively reviewed a series of thoracolumbar injury case studies. Thirteen case studies, including images, were distributed to STSG members for individual, independent evaluation of the following three criteria: (1) diastasis of the facet joints on CT; (2) posterior edema-like signal in the region of PLC components on sagittal T2-weighted fat saturation (FAT SAT) MRI; and (3) disrupted PLC components on sagittal T1-weighted MRI. Interrater agreement on the presence or absence of each of the three criteria in each of the 13 cases was assessed. RESULTS: Absolute interrater percent agreement on diastasis of the facet joints on CT and posterior edema-like signal in the region of PLC components on sagittal T2-weighted FAT SAT MRI was similar (agreement 70.5%). Interrater agreement on disrupted PLC components on sagittal T1-weighted MRI was 48.9%. Facet joint diastasis on CT was the most reliable indicator of PLC disruption as assessed by both Cohen's kappa (kappa = 0.395) and intraclass correlation coefficient (ICC 0.430). CONCLUSIONS: The interrater reliability of assessing diastasis of the facet joints on CT had fair to moderate agreement. The reliability of assessing the posterior edema-like signal in the region of PLC components was lower but also fair, whereas the reliability of identifying disrupted PLC components was poor. PMID- 17909930 TI - Quantitative gait analysis of patients with bilateral hip osteoarthritis excluding the influence of walking speed. AB - BACKGROUND: The purpose of this study was to investigate gait characteristics of patients with bilateral hip osteoarthritis (BHO) independent of walking speed. METHODS: We measured gait parameters in 12 BHO patients during free walking activities and in 12 normal adults during both free walking and slow walking activities using a three-dimensional computerized gait analysis system. RESULTS: Patients with BHO had a lower walking speed, step length, and cadence than normal subjects during free walking. When compared with normal subjects walking at a slow speed, the walking speed difference among BHO patients disappeared, although BHO patients retained a relatively high cadence. Kinematic and kinetic factor analysis of BHO patients at free speed compared to normal subjects walking at a slow speed showed a forward-tilted pelvic angle in the BHO patients that dropped to that of the ipsilateral side during the stance phase. The peak extension and abduction angle of the hip and the peak abduction moment of the hip were all low, whereas the peak generation power of the ankle was high in BHO patients. CONCLUSIONS: Gait characteristics of patients with BHO, independent of walking speed, were as follows: (1) increased cadence and ankle generation power; (2) reduced step width, hip extension, and abduction angle as well as a lower hip abduction moment; (3) maintained forward tilting of the pelvis during gait cycle; and (4) appearance of a dropped pelvis during the stance phase. PMID- 17909929 TI - JOA back pain evaluation questionnaire: initial report. AB - BACKGROUND: There is no widely accepted objective evaluation for lumbar spine disorders. New outcome measures should be patient-oriented and should measure symptoms and self-reported functional status in multiple dimensions. The aim of this study was to identify items to be included in the disease-specific quality of life (QOL) questionnaire for the assessments of patients with lumbar spine disorders. METHODS: The draft of the QOL questionnaire that consisted of a total of 60 items, including 24 items derived from the Japanese version of the Roland Morris Disability Questionnaire (RDQ) and 36 items derived from the Japanese version of Short Form 36 (SF-36), were administered to patients and controls. After obtaining written informed consent, the following data were collected from the patient group (n = 328) and the control group (n = 213): (1) background characteristics, including age, diagnosis, Japanese Orthopaedic Association (JOA) score, and finger to floor distance; (2) responses to the questionnaire; (3) the identification rate by discrimination analysis to select the candidates for adoption and by adopting explanatory variables. The items to be excluded were determined by examining the explanatory variables, which were selected after the discrimination analysis, by setting the candidate to-be-excluded items as an objective variable. RESULTS: Based on the distribution of the responses, two items, RDQ-15 and RDQ-19, were excluded. From the results of the correlation coefficient calculation for each question in the patient group, 33 items were excluded and 27 candidate items were adopted. Based on the adoption explanatory variable used in the discrimination analysis, 25 of the 27 candidate items for adoption were accepted. CONCLUSIONS: This study identified the 25 specific questionnaire items that should be included in the questionnaire to evaluate QOL of patients with various lumbar spine disorders. PMID- 17909931 TI - Successful results of minimally invasive surgery for comminuted supracondylar femoral fractures with LISS: comparative study of multiply injured and isolated femoral fractures. AB - BACKGROUND: The aim of this prospective study was to compare the outcomes of distal femoral fractures treated by the Less Invasive Stabilization System (LISS) in multiply injured and isolated fracture cases. METHODS: This study comprised 26 patients (16 men, 10 women), who had 27 distal femoral fractures. Patients were divided into two groups; multiple injuries (group I) or isolated distal femoral fracture (group II). The average Injury Severity Score of group I was 26.7. Operations were performed according to biological fixation principles in a submuscular manner. No bone grafting was performed to enhance the healing. The cases were evaluated based on the criteria of Schatzker-Lambert and the modified Hospital for Special Surgery (HSS) scoring system. RESULTS: The mean follow-up period was 25.8 months. Union was achieved in all cases. Two patients in group I required debridement procedures due to deep infection. One of them healed completely but the other did not. The average range of knee motion of groups I and II at the last control were 112.8 degrees and 121.8 degrees , respectively. The mean modified HSS scores were 73.9 and 79.9, respectively. There was no significant difference in the HSS scores or the range of knee motion. The time to full weight bearing was longer in group I owing to the concomitant injuries. CONCLUSIONS: We concluded that LISS is a useful method for comminuted supracondylar fractures with multiple trauma patients as the results showed no significant differences when compared with those of patients with isolated femoral fractures. PMID- 17909932 TI - Poor results of the cementless total hip arthroplasty with a nonporous coated acetabular component: AcSys Shearer Cup. AB - BACKGROUND: This study followed patients for a minimum of 7 years after primary total hip arthroplasty using cementless acetabular components and evaluated their outcomes. METHODS: We followed 73 patients (75 hips), who had undergone total hip arthroplasty with cementless nonporous coated acetabular components (3M AcSys Shearer Cup) for a mean of 9.8 years (range 7-13 years). There were 61 women and 12 men with a mean age of 53 years (range 27-69 years) at surgery. The diagnosis was primary osteoarthritis in 9 hips, osteoarthritis secondary to developmental dysplasia in 58 hips, osteonecrosis of the femoral head in 6 hips, and rheumatoid arthritis in 2 hips. RESULTS: Three cups were revised because of aseptic loosening, and one cup was revised following removal of the prosthesis due to deep infection. Radiographic loosening was observed in 22 hips at the latest follow-up. The survival rate at 10 years was 94.7% with revision as the endpoint and 72% with radiographic loosening as the endpoint. The Merle d'Aubigne and Postel hip score showed significant improvement postoperatively and was maintained well even in cases showing radiographic loosening. CONCLUSIONS: The intermediate radiological results with the AcSys Shearer Cup were unsatisfactory because of the high loosening rate, although the revision rate was low. The nonporous outer surface and the poor fixation mechanism between the metal shell and liner may have contributed to the high failure rate. Regular radiological review is recommended when this cup is used because early loosening is often painless. PMID- 17909933 TI - Treatment of lower limb deformities and limb-length discrepancies with the external fixator in Ollier's disease. AB - BACKGROUND: In this study, we addressed two questions on the treatment for Ollier's disease: (1) how much callus formation occurs when an osteotomy is performed intralesionally and (2) how is the stability of the wires and half-pins that are inserted intralesionally. METHODS: Four children with Ollier's disease underwent treatment of 12 lower limb segments using distraction osteogenesis until completion of their growth. All osteotomies were performed at the centers of the deformities, resulting in a total of seven osteotomies performed intralesionally. RESULTS: Full correction of the deformity and full restoration of length were achieved in all cases, but a residual limb-length discrepancy of <10 mm remained. The mean external fixation index in the intralesional distraction osteogenesis group was 39.7 days/cm versus 30.8 days/cm in the extralesional distraction osteogenesis group. Conversion from abnormal cartilage to normal regenerate bone was seen in only one segment. Although approximately two-thirds of the wires and half-pins were inserted intralesionally, in all but one case (in which an iatrogenic fracture occurred) the wires and half-pins were well stabilized throughout the external fixation period. CONCLUSIONS: Although deformity and limb-length discrepancies due to Ollier's disease were successfully resolved by distraction osteogenesis, enchondroma may arise in distracted calluses when osteotomized intralesionally. However, the stability of the external fixator was sufficient to lengthen limbs and correct deformities even when wires and half-pins were inserted intralesionally. PMID- 17909934 TI - In vivo kneeling biomechanics after posterior stabilized total knee arthroplasty. AB - BACKGROUND: Kneeling is one of the activities sought by patients after total knee arthroplasty (TKA). This study investigated the six degrees of freedom (DOF) kinematics and three-dimensional (3D) contact during weight-bearing kneeling. METHODS: A total of 16 South Korean female patients (22 knees) after posteriorly stabilized (PS) TKA (LPS-Flex) were randomly recruited and had the same surgeon. The patients were imaged using a dual fluoroscopic technique while they kneeled from initial to maximum flexion. The acquired images and 3D models were then used to recreate the in vivo pose of the components. Contact was determined by locating the surface intersections in the tibiofemoral and cam/post (between the femoral cam and tibial post) articular compartments. RESULTS: Patients flexed, on average, from 107.3 degrees to 128.0 degrees during the kneeling activity. Changes in kinematics included 1.0 mm of proximal, 0.9 mm of medial, and 7.6 mm of posterior translation and 1.7 degrees of varus rotation (P < 0.04). A difference in internal tibial rotation was not detected. Articular contact moved posteriorly by 5.9 mm and 6.4 mm in the medial and lateral compartments, respectively. Contact also moved medially by 3.2 mm and 5.8 mm in the medial and lateral compartments. A decrease in articular contact was observed in both condyles, and lateral condylar lift-off increased with flexion (P = 0.0001). More than 80% of the patients demonstrated cam/post engagement, which always occurred in the distal portion of the post. CONCLUSIONS: In this patient cohort, the knee joint was constrained during the weight-bearing activity such that femoral subluxation and dislocation were not observed. Furthermore, posterior cam/post engagement occurred only in the distal portion of the tibial post, which may improve the longevity of the post. The tibiofemoral and cam/post articular contact data presented in this study further suggest that kneeling may be performed by patients after clinically successful PS TKA who feel comfortable with the activity and are free of pain. PMID- 17909935 TI - Comparison of the cytokine-induced migratory response between primary and subcultured populations of rat mesenchymal bone marrow cells. AB - BACKGROUND: The aim of this study was to investigate the phenotypic differences of primary rat mesenchymal bone marrow cells (MBMCs) and subcultured cells, the influence of subculture and cell density on the cellular phenotypes, and the difference in the migratory responses of these cells to cytokines. METHODS: MBMCs were isolated from 8-week-old Wistar rats, and the cells were cultured for 1 week (passage 0, P0) or 3 weeks (P0-3W). P0 cells were subcultured for 1 week (P1). P1 cells were subcultured at several cell densities for 1 week (P2). Cell size and granularity were analyzed by flow cytometry. The gene expression characteristics of these cells were analyzed by reverse transcription polymerase chain reaction. Cell migration to bone morphogenetic protein-2 (BMP-2), fibroblast growth factor 2 (FGF-2), and platelet-derived growth factor-bb (PDGF-bb) was evaluated using a Boyden chamber. RESULTS: Three morphologically distinct populations in P0 and two in P2 were detected. The levels of human rapidly self-renewing cell-related marker genes in P0 were more highly expressed than in P2. Mesenchymal stem cell associated markers were expressed at the same level in P0 and P2. The gene expression levels of immature oligodendrocyte precursor cell markers in P0 were higher than those in P2, whereas those of smooth muscle cell markers and osteoblastic cell markers in P0 were lower than those in P2. Subculture decreased the gene expression levels of human rapidly self-renewing cell-associated markers. Cell migration of P0 cells was stimulated by PDGF-bb but not by BMP-2 or FGF-2. In contrast, PDGF-bb, BMP-2, and FGF-2 all stimulated cell migration of P2. CONCLUSION: The types of cells in populations of primary and subcultured rat MBMCs were different, and the distribution of each cell population appeared to be changed by the culture conditions. The cell migration effect by PDGF-bb, BMP-2, and FGF-2 differed between the primary and subcultured MBMCs. PMID- 17909936 TI - Proximal interphalangeal joint silicone arthroplasty for posttraumatic arthritis. PMID- 17909937 TI - Gossypiboma (foreign body granuloma) mimicking a soft tissue tumor with hip hemiarthroplasty. PMID- 17909938 TI - Angioleiomyoma overlying the Achilles tendon. PMID- 17909939 TI - Ilizarov deformity correction of the lower limbs in Ellis-van Creveld syndrome. PMID- 17909940 TI - Nontraumatic subluxation of the hip after spine surgery for scoliosis in a patient with von Recklinghausen's disease. PMID- 17909941 TI - Conventional MRI cannot predict survival in childhood diffuse intrinsic pontine glioma. AB - Diffuse intrinsic pontine glioma (DIPG) of childhood has a dismal prognosis. Clinical trials of new agents are vital and it is essential that the correct endpoints and disease assessments are chosen. A retrospective review of magnetic resonance imaging (MRI) scanning in a pure population of DIPG was undertaken. Baseline diagnostic MRI findings included; local tumour extension in upper medulla (74%) or midbrain (62%), metastatic disease (3%), basilar artery encasement (82%), necrosis (33%), intratumoural haemorrhage (26%), hydrocephalus (23%) and dorsal exophytic component (18%). Post-treatment MRI scans demonstrated increases in; leptomeningeal metastatic disease (16%), cystic change/necrosis (48%), enhancement (72%) and intratumoural haemorrhage (32%). Response rates were calculated according to both RECIST (4%) and WHO (24%) criteria. No MRI parameter in either the diagnostic or response scans had prognostic significance. We recommend that currently primary endpoints for DIPG clinical trials should be overall or possibly progression free survival and that new advanced functional imaging techniques should be explored as possible surrogate markers for novel therapy activity rather than conventional MRI response criteria. PMID- 17909942 TI - CSF IgH gene rearrangement analysis in isolated post-transplant lymphoproliferative disorder of the central nervous system. AB - Isolated post-transplant lymphoproliferative disorder (PTLD) of the central nervous system is rare and its diagnosis can be challenging. A biopsy is usually required in order to distinguish the disease from opportunistic infections. We present a case in whom immunoglobulin heavy chain gene rearrangement analysis of cerebrospinal fluid (CSF) was instrumental in establishing the diagnosis. PMID- 17909943 TI - C-Terminal truncation affects subunit exchange of human alphaA-crystallin with alphaB-crystallin. AB - In human lenses, C-terminal cleavage of alphaA-crystallin at residues 172,168, and 162 have been reported. The effect of C-terminal truncation of alphaA crystallin on subunit exchange and heterooligomer formation with alphaB crystallin and homooligomer formation with native alphaA-crystallin is not known. We have conducted fluorescence resonance energy transfer studies which have shown that the rates of subunit exchange of alphaA(1-172 )and alphaA(1-168 )with alphaB wt were two-fold lower than for alphaA-wt interacting with alphaB-wt. The subunit exchange rate between alphaA(1-162) and alphaB-wt was six-fold lower. These data suggest that cleavage of the C-terminal residues could significantly affect heterooligomerization. On the other hand, the subunit exchange rates between alphaA-wt and the truncated alphaA-crystallins were either unchanged or only slightly decreased, which suggest that homooligomerization may not be significantly influenced by C-terminal truncation. The main conclusion from this study is that cleavage of C-terminal residues of alphaA-crystallin including the nine residues of the flexible tail is expected to significantly affect the formation of heteroaggregates. Reconstitution experiments showed that the presence of an intact C-terminus is essential for the formation of fully integrated heteroaggregates with equal proportion of alphaA and alphaB subunits. PMID- 17909944 TI - Metformin promotes isolated rat liver mitochondria impairment. AB - Metformin, a drug widely used in the treatment of type 2 diabetes, has recently received attention due to the new and contrasting findings regarding its effects on mitochondrial function. In the present study, we evaluated the effect of metformin in isolated rat liver mitochondria status. We observed that metformin concentrations > or =8 mM induce an impairment of the respiratory chain characterized by a decrease in RCR and state 3 respiration. However, only metformin concentrations > or =10 mM affect the oxidative phosphorylation system by decreasing the mitochondrial transmembrane potential and increasing the repolarization lag phase. Moreover, our results show that metformin does not prevent H(2)O(2) production, neither protects against lipid peroxidation induced by the pro-oxidant pair ADP/Fe(2+). In addition, we observed that metformin exacerbates Ca(2+)-induced permeability transition pore opening by decreasing the capacity of mitochondria to accumulate Ca(2+ )and increasing the oxidation of thiol groups. Taken together, our results show that metformin can promote liver mitochondria injury predisposing to cell death. PMID- 17909945 TI - Modulation of calcification of vascular smooth muscle cells in culture by calcium antagonists, statins, and their combination. AB - BACKGROUND: Vascular calcification is an organized process in which vascular smooth muscle cells (VSMCs) are implicated primarily. The purpose of the present study was to assess the effects of calcium antagonists and statins on VSMC calcification in vitro. METHODS: VSMC calcification was stimulated by incubation in growth medium supplemented with 10 mmol/l beta-glycerophosphate, 8 mmol/l CaCl(2), 10 mmol/l sodium pyruvate, 1 micromol/l insulin, 50 microg/ml ascorbic acid, and 100 nmol/l dexamethasone (calcification medium). Calcification, proliferation, and apoptosis of VSMCs were quantified. RESULTS: Calcium deposition was stimulated dose-dependently by beta-glycerophosphate, CaCl(2), and ascorbic acid (all P < 0.01). Addition of amlodipine (0.01-1 micromol/l) to the calcification medium did not affect VSMC calcification. However, atorvastatin (2 50 micromol/l) stimulated calcium deposition dose-dependently. Combining treatments stimulated calcification to a degree similar to that observed with atorvastatin alone. Both atorvastatin and amlodipine inhibited VSMC proliferation at the highest concentration used. Only atorvastatin (50 micromol/l) induced considerable apoptosis of VSMCs. CONCLUSION: In vitro calcification of VSMCs is not affected by amlodipine, but is stimulated by atorvastatin at concentrations > or =10 micromol/l, which could contribute to the plaque-stabilizing effect reported for statins. PMID- 17909946 TI - Analysis of the suitability of calreticulin inducible HEK cells for adhesion studies: microscopical and biochemical comparisons. AB - Calreticulin is a Ca(2+)-buffering ER chaperone that also modulates cell adhesiveness. In order to study the effect of calreticulin on the expression of adhesion-related genes, we created a calreticulin inducible Human Embryonic Kidney (HEK) 293 cell line. We found that fibronectin mRNA and both intra- and extra-cellular fibronectin protein levels increased following calreticulin induction. However, despite this increase in fibronectin, HEK293 cells did not assemble an extracellular fibrillar fibronectin matrix regardless of the level of calreticulin expression. Furthermore, HEK293 cells exhibited a poorly organized actin cytoskeleton, did not have clustered fibronectin receptors at the cell surface, and did not form focal contacts. This likely accounts for the lack of fibronectin matrix deposition by these cells regardless of calreticulin expression level. Vinculin abundance did not appreciably increase upon calreticulin induction and the level of active c-Src, a regulatory kinase of focal contacts, was found to be abundant and unregulated by calreticulin induction in these cells. The inability to form stable focal contacts and to commence fibronectin fibrillogenesis due to high c-Src activity may be responsible for the poor adhesive phenotype of HEK 293 cells. Thus, we show here that HEK293 cells are not suitable for microscopical studies of cell-substratum adhesions, but are best suited for biochemical studies. PMID- 17909947 TI - Thyroid hormone regulates the expression of SNAP-25 during rat brain development. AB - Thyroid hormones are major regulators of postnatal brain development. Thyroid hormones act through nuclear receptors to modulate the expression of specific genes in the brain. We have used microarray analysis to identify novel responsive genes in 14-day-old hypothyroid rat brains, and discovered that synaptosomal associated protein of 25 kDa (SNAP-25) was one of the thyroid hormone-responsive genes. SNAP-25 is a presynaptic plasma membrane protein and an integral component of the vesicle docking and fusion machinery mediating secretion of neurotransmitters and is required for neuritic outgrowth and synaptogenesis. Using microarray analysis we have shown that SNAP-25 was down-regulated in the hypothyroid rat brain compared with the age-matched controls. Real-time RT-PCR and western blotting analysis confirmed that SNAP-25 mRNA and protein levels decreased significantly in the developing hypothyroid rat brain. Our data suggest that in the developing rat brain, SNAP-25 expression is regulated by thyroid hormone, and thyroid hormone deficiency can cause decreased expression of SNAP-25 and this may on some level account for the impaired brain development seen in hypothyroidism. PMID- 17909948 TI - Alteration of mitochondrial oxidative capacity during porcine preadipocyte differentiation and in response to leptin. AB - Mitochondrial apparatus is a fundamental aspect in cell, serving for amino acid biosynthesis, fatty acid oxidation (FAO), and ATP production. In this article, we investigated the change of mitochondrial oxidative capacity during porcine adipocyte differentiation and in response to leptin. Rhodamine 123 staining analysis showed about 2-fold increase of mitochondrial membrane electric potential in differentiated adipocyte in comparison with preadipocyte. The mRNA expression of Cytochromes c (Cyt c), carnitine palmitoyltransferase 1 (CPT1), and malate dehydrogenases (MDH) increased markedly (P < 0.05), but that of UCP2 decreased (P < 0.05). Moreover PGC-1alpha and UCP3 was very low and showed no changes during the adipocyte differentiation. The protein expression of Cyt c and the enzyme activity of Cytochrome c oxidase (COX) increased with preadipocyte differentiation, but cellular ATP level decreased. Furthermore, at the level of 10 and 100 ng/ml leptin not only selectively increased the gene expression of PGC 1alpha, CPT1, Cyt c, UCP2, and UCP3 (P < 0.05), but also enhanced COX enzyme activity which related to mitochondrial FAO. There is no change of Mitochondrial membrane electric potential and ATP level in cell treated by leptin. These results suggested Mitochondrial is not only critical in FAO, but also play an important role in adipogenesis. PMID- 17909949 TI - Cross-cultural adaptation, reliability, and validity of the work role functioning questionnaire to Brazilian Portuguese. AB - INTRODUCTION: The study objectives were to translate and adapt the Work Role Functioning Questionnaire (WRFQ) into the Brazilian Portuguese language and evaluate its reliability in patients experiencing musculoskeletal disorders. METHODS: The cross-cultural adaptation was performed according to the internationally recommended methodology, using the following guidelines: translation, back-translation, revision by a committee, and pretest. At first, the questionnaire was independently translated by two bilingual translators, who had Portuguese as their mother language. Subsequently, two other translators whose mother language was English did the back-translation. A committee composed of five specialists revised and compared the translations obtained, developing the final version for pretest application. The pretest was carried out with 30 patients experiencing musculoskeletal disorders. Psychometric properties were evaluated by administering the questionnaire to 105 subjects with musculoskeletal disorders and receiving physical therapy treatment. The reliability was estimated through stability and homogeneity assessment. The construct validity was tested comparing subjects experiencing musculoskeletal disorders to healthy workers. RESULTS: The results indicated good content validity and internal consistency (Cronbach alpha = 0.95). Cronbach alpha for each scale was >0.85, except for the social demand scale. The Intraclass Correlation Coefficient for the test-retest reliability was satisfactory for mental demands (ICC = 0.68) and excellent for the others (0.82-0.91). In relation to the construct validity, the mean score obtained for each scale was lower for physical, work scheduling, and output demands in the subjects with musculoskeletal disorders. There was a significant difference (p < 0.001) between the groups in comparison to work scheduling, physical, and output demands. CONCLUSIONS: The data showed that the cross cultural adaptation process was successful and the adapted instrument demonstrated psychometric properties making it reliable to use in Brazilian culture. PMID- 17909951 TI - Spectroscopic Properties of Er3+/Yb3+ -codoped PbO-Bi2O3-Ga2O3-GeO2 glasses. AB - We investigate the spectroscopic properties of the 1.5-microm emission from the (4)I(13/2)-->(4)I(15/2) transition of Er(3+) ions in PbO-Bi(2)O(3)-Ga(2)O(3) GeO(2) glasses for applications in broadband fiber amplifiers. The measured emission peak locates at 1,532 nm with a full width at half-maximum of approximately 45 nm. The glasses exhibit a large stimulated emission cross section of 0.89 x 10(-20) cm(2) and a large FWHM x sigma(e)(peak) product of 40.0. Infrared-to-green upconversion occurs simultaneously upon excitation of the 1.5-microm emission with a commercially available 980 nm laser diode. The green upconversion intensity has a quadratic dependence on incident pump laser power, indicating a two-photon process. Energy transfer processes and nonradiative phonon-assisted decays could account for the population of the (2)H(11/2) of Er(3+). The results indicate the possibility towards the development of lead bismuth-gallate-germanate based glasses as photonics devices. PMID- 17909950 TI - Subjective quality of life according to work status following interdisciplinary work rehabilitation consequent to musculoskeletal disability. AB - INTRODUCTION: Individualized subjective quality of life (ISQoL) is the appraisal of quality of life according to personal values, desired goal attainment and life priorities. "Gap" is a way to operationalize ISQoL. ISQoL is rarely measured by interdisciplinary work rehabilitation (IWR) programs attended by the musculoskeletal disorder (MSD) population. These programs commonly measure pain intensity, physical capacity, perceived disability, distress and return to work. OBJECTIVES: The aims of this study were to compare ISQoL according to work status and reference values and to explore the relationships between ISQoL and common IWR measures. METHODS: Six months after completing an IWR program, 40 working and 31 not-working participants completed questionnaires documenting work status, pain, ISQoL gap, health-related quality of life (SF-36, PCS and MCS), perceived disability and distress. RESULTS: No significant difference in global ISQoL gap was found between working and not-working participants. When compared to reference values considerable variability exists but globally, for both groups, ISQoL gap scores were below average. The following clinical variables were related to global ISQoL (P < 0.05): pain (r = -0.42), PCS (r = -0.37), MCS (r = 0.56), perceived disability (r = 0.37) and distress (r = 0.61). High distress, present in both groups, explains 38% of the global ISQoL gap variance and PCS adds 4%. CONCLUSION: Following IWR programs for the chronic MSD population, global ISQoL gap is not related to work status. The use of a client-centered interactive computerized measure of ISQoL reveals that domains related to emotional well-being are likely the most problematic for the persistently disabled MSD population. PMID- 17909952 TI - Improving visible light sensitization of luminescent europium complexes. AB - The synthesis and characterization of the new ligands L(1), L(2) and L(4) are described with the series of four europium complexes of formula [EuL(n)(TTA)(3)] in which TTA refers to 2-thenoyltrifluoroacetonate and L(n) to tridentate ligands with nitrogen containing heterocyclic structure, such as a 2,6-bis(3-methyl pyrazolyl)-4-(p-toluyl-ethynyl)-triazine for L(1), or terpyridines functionalized at the 4' position by a phenyl-vinylene for L(2), a p-dimethylamino-phenylene for L(3), or a p-aminophenyl-ethynylene for L(4). The spectroscopic properties of the ligands and of the complexes are studied by means of UV-Vis absorption spectroscopy, as well as steady-state and time-resolved luminescence spectroscopy. All complexes display europium centred luminescence upon ligand excitation. Careful examination of the excitation spectra revealed differences in the ligand based sensitization efficiencies. For complexes of L(1) and L(2), excitation of europium is mainly achieved through the TTA moieties and the photo physical studies on [EuL(1)(TTA)(3)] evidenced a weaker coordination of the bispyrazolyltriazine tridentate ligand, resulting from a partial decomplexation upon dilution. Complexes of L(3) and L(4) display intense excitation through the tridentate units, which extend down to 460 nm in the visible region. In the case of L(3), selective excitation reveals the presence of a ligand-centred emission band at 520 nm which is likely ascribed to a L(3) centred charge transfer state. PMID- 17909953 TI - A multicenter, prospective, open label, historically controlled clinical trial to evaluate efficacy and safety in primary immunodeficiency diseases (PID) patients of Flebogamma 5% DIF, the next generation of Flebogamma. AB - BACKGROUND: Flebogamma 5% dual inactivation and filtration (DIF) is the next generation of Flebogamma. Flebogamma was first licensed in 1992. The new preparation features additional viral inactivation and removal steps to enhance safety margins. OBJECTIVE: The purpose of this study was to evaluate the efficacy, safety, and pharmacokinetics of Flebogamma 5% DIF for immunoglobulin replacement therapy in primary immunodeficiency diseases (PID). METHODS: Flebogamma 5% DIF was administered at seven clinical sites to 46 subjects with well-defined primary immunodeficiency diseases at a dose of 300-600 mg/kg every 21-28 days for 12 months. RESULTS: The calculated serious bacterial infection rate was 0.021/subject/year. The incidence of adverse events considered potentially related to Flebogamma 5% DIF during or within 72 h after completing an infusion was approximately 10%. The half-life in serum of the administered IgG was around 31 days. CONCLUSIONS: Flebogamma 5% DIF is efficacious and safe, has adequate pharmacokinetic properties, is well-tolerated and maintains the profile of Flebogamma 5% for the treatment of primary humoral immunodeficiency diseases. PMID- 17909955 TI - Venous valves: unseen obstructions to coronary access. PMID- 17909954 TI - BCG-induced rabbit alveolar macrophages are endowed with strengthened antioxidant metabolic pathways. AB - Following i.v. BCG infection, a new population of macrophages are recruited in the rabbit lung. These macrophages, known as activated macrophages, substitute the resident macrophages and can play a key role in the defence against mycobacteria. We report here that BCG-activated alveolar macrophages are equipped with a more active hexose monophosphate pathway, which can maintain an optimal intracellular concentration of NADPH and GSH, and allow to produce mycobactericidal free radicals and to become resistant to mycobacterium-induced programmed cell death. These findings suggest that sustaining the anti-oxidant properties of macrophages could represent a candidate process to be considered as a good therapeutic target in fighting Mycobacterium spp infections. PMID- 17909957 TI - The effect of the phytoestrogen genistein on myocardial protection and preconditioning in hypercholesterolemia. PMID- 17909956 TI - Angiotensin II type 1 receptor inhibition is associated with reduced tachyarrhythmia-induced ventricular interstitial fibrosis in a goat atrial fibrillation model. AB - BACKGROUND: Using a goat animal model, we tested the hypothesis that angiotensin II inhibition reduces fibrotic degeneration of both the atrial and ventricular myocardium as well as AF induction susceptibility. METHODS: We studied three groups of five goats over a 6-month period. The study animals in the first two groups were implanted with a pacemaker capable of maintaining AF with burst pacing. Additionally, in one group, goats were administered candesartan (AF+candesartan group). The third group (SR group) of animals served as control. Animals were tested for AF induction on day 0, 1, 30, 90 and 180. A "Vulnerability Index" (VI) for AF induction was calculated, defined as the ratio of total time in AF per number of bursts needed to induce sustained AF, in each session. At the end of the study, all four heart chambers were examined and fibrosis quantified. RESULTS: Both AF goat groups developed cardiomegaly due to tachy-cardiomyopathy. Although, the VI was significantly increased in AF group over time (28.8+/-43 to 284.7+/-291, p=0.045), this was not the case for AF+candesartan group (30.3+/-40 to 170.8+/-243, p=0.23). Histology revealed a significant increase of fibrous tissue in goats with induced AF, noticeable in all four heart chambers, compared to controls. However, the degree of fibrosis was significantly lower in AF animals on candesartan. CONCLUSIONS: Our study demonstrated a beneficial effect of angiotensin II inhibition on tachyarrhythmia induced ventricular fibrosis. It is also consistent with previous studies indicating a reduction in burst-induced AF susceptibility in goats and confirms the favorable effects in atrial structural remodeling. PMID- 17909958 TI - Disgust and contamination sensitivity in vaginismus and dyspareunia. AB - This study examined the potential role of disgust propensity and contamination sensitivity in vaginismus. Women suffering from vaginismus (n = 20) or dyspareunia (n = 22), and a group of women without sexual complaints (n = 30) completed self report measures indexing their (1) general dispositional disgust propensity, and (2) sensitivity for (ideational) contamination by sexual stimuli as a function of its source (self, partner, unknown). In support of the idea that disgust may be involved in vaginismus, women with vaginistic complaints displayed a generally enhanced dispositional disgust propensity. The sensitivity for contamination by sexual stimuli did not vary across groups. However, especially when the source was the participant's partner, the willingness ratings might have been influenced by demand and may, therefore, not accurately reflect participant's actual sensitivity for contamination by sexual stimuli. Future studies using more implicit or behavioral measures are necessary to more definitely test the role of disgust in vaginismus. PMID- 17909960 TI - Sexual pleasure and condom use. AB - The purpose of the present study was to determine whether sexually-experienced individuals' pleasure ratings for protected and unprotected vaginal intercourse would be related to actual condom use. College participants (80 women and 35 men, M age = 22.29 years) who reported engaging in vaginal intercourse in the past 3 months completed a questionnaire that assessed their perceptions of the pleasurability of unprotected and condom-protected vaginal intercourse and their own sexual behaviors. Both women and men rated unprotected vaginal intercourse as more pleasurable than protected vaginal intercourse. However, men's pleasure ratings for unprotected vaginal intercourse were higher than women's. Furthermore, men and women's pleasure ratings for condom-protected intercourse were correlated with their actual condom use behaviors. Men's "pleasure decrement" scores indicated a significantly greater reduction in pleasure ratings between unprotected and protected intercourse than women's scores. Men who perceived a larger decrease in pleasure between unprotected and protected intercourse were less likely to have used condoms in the past 3 months than those who perceived a smaller decrease in pleasure. The results provide evidence that many people believe that condoms reduce sexual pleasure and that men, in particular, who believe that condoms decrease pleasure are less likely to use them. Condom promotion campaigns should work to emphasize the pleasure-enhancing aspects of condom use. PMID- 17909961 TI - Work barriers in the context of pathways to the employment of welfare-to-work clients. AB - The ability of welfare-to-work clients to leave the welfare rolls and stay in the labor force is often limited by the work barriers they face. Using a sample of 1,404 female welfare-to-work clients we first examined the structure of work barriers and then tested their contribution to current work status in the context of a structural equation model that incorporated other central pathways to employment. Whereas work barriers included diverse factors ranging from lack of transportation to low quality jobs, they were shown to constitute a uni dimensional construct. Furthermore, work barriers had a net adverse effect on employment outcomes, controlling for job search self-efficacy and employment intention. We conclude with discussion of implications for the development of welfare-to-work programs and interventions that target low-income women. PMID- 17909962 TI - The significance of employment for chronic stress and psychological distress among rural single mothers. AB - Considerable research has found elevated levels of stress and psychological distress among single mothers. However, little research has addressed the relevance of employment for stress processes in this population, and few studies have focused on links between employment, stress, and mental health among single mothers living in rural areas. Based on a probability sample of 508 single mothers age 18-39 living in rural Northern New England, this study: (1) documents variations in chronic stress and psychological distress for employed and unemployed single mothers, (2) considers the extent to which different forms of chronic stress mediate or explain the employment-distress relationship, and (3) examines whether employment status modifies associations between chronic stress and psychological distress. Findings indicated that rural single mothers who were employed reported significantly less financial stress, childcare stress, and rural residence stress, relative to mothers who were not employed, independent of variety of other factors. Employed mothers also experienced significantly less psychological distress which was partially mediating by their lower financial stress. However, there was also a statistical interaction between employment status and financial stress such that the harmful effect of financial stress on well-being was greater for employed women. Implications of the findings are discussed. PMID- 17909963 TI - The multidimensional nature of social cohesion: psychological sense of community, attraction, and neighboring. AB - Items from the Neighborhood Cohesion Instrument were completed by 1,732 individuals from a random sample of households in 20 rural communities across Canada during the summer of 2001. Confirmatory factor analysis of the NCI items based on polychoric correlations and weighted least squares estimation found three underlying latent variables. Although items were related to the three latent variables in a somewhat different manner than they were in Buckner's original study, the same three latent variables were evident, providing convincing evidence that social cohesion has at least three subscales: psychological sense of community, neighboring, and attraction. Correlations between subscales were relatively high (between .67 and .87). Intraclass correlation coefficients for the three scales were .115, .127, and .112. In addition, the community means differed on different subscales in a manner that related to recognized characteristics of the communities. Thus the subscales are appropriate measures for both individuals and communities and can be recommended for further research on social cohesion. PMID- 17909964 TI - Genetic polymorphisms in uridine diphospho-glucuronosyltransferase 1A1 and breast cancer risk in Africans. AB - The UDP-glucuronosylatransferase 1A1 (UGT1A1) gene is involved in the metabolism of estrogen and detoxification of potential carcinogens. The number of TA repeats in the promoter region of UGT1A1 has been linked to breast cancer risk, but results varied by race. We performed a comprehensive assessment of genetic polymorphisms in the UGT1A1 gene, and examined these polymorphisms and TA repeats in relation to breast cancer risk in a case-control study in Nigeria. 512 breast cancer cases and 226 community controls were genotyped for UGT1A1. Compared with high-activity TA repeat genotypes, the odds ratios (OR) for low-activity and moderate-activity genotypes were 0.47 (95% confidence interval CI, 0.26-0.83) and 0.64 (95% CI, 0.39-1.06), respectively, in premenopausal women (P = 0.009 for trend), but no association was observed in postmenopausal women (P = 0.24). The effect of TA repeats was also differentiated by age: the OR was 0.39 (95% CI 0.21 0.71) for low-activity genotypes and 0.58 (95% CI 0.33-1.00) for moderate activity genotypes in women <45 years old (P = 0.002 for trend), but no association was observed in women >or=45 years old (P = 0.15). Haplotype analysis showed that UGT1A1 haplotypes were highly diverse with blocked structures. We found a specific haplotype in block 2 that was significantly associated with a 2.1-fold elevated risk (95% CI 1.05-4.39; P = 0.04). In contrast with previous studies, we found low-activity TA repeat alleles were protective against breast cancer among premenopausal indigenous Africans, suggesting that the role of UGT1A1 in breast cancer development may vary by population, presumably due to different environmental and genetic modifier effects. PMID- 17909965 TI - Serglycin and secretion in human monocytes. AB - The human monocytic cell line U-937 has been widely used as a model system for human monocytes. The subclone U-937-B has been adapted to serum-free conditions. This particular U-937 clone and its parent clone U-937-1 were used to investigate the role of the proteoglycan serglycin in human monocytes. For this purpose cells were treated with hexyl-beta-D-thioxyloside to abrogate proteoglycan expression. U-937-B cells expressed and secreted exclusively chondroitin sulphate proteoglycans, and after treatment with this xyloside they only expressed and released free chondroitin sulphate chains. Western blotting showed that serglycin core protein was present in conditioned medium of control cells, but absent in medium from xyloside-treated cells. Also, serglycin core protein could be detected in the cell fractions of control cells, but not in the cell fractions from xyloside-treated cells. Furthermore, less proteoglycan-associated proteins could be detected in medium from cells incubated with xyloside, suggesting that the absence of secreted sergycin affects the secretion of such proteins. Cells incubated in the presence of xyloside were analyzed by transmission electron microscopy and shown to contain numerous large empty vesicles. The lack of serglycin, the dominant proteoglycan in U-937 monocyte-like cells, consequently, leads to effects on vesicle formation and secretion of some low molecular weight proteins, suggesting that this particular proteoglycan is of importance for secretory processes in human monocytes. PMID- 17909966 TI - Characterization of anticoagulant heparinoids by immunoprofiling. AB - Heparinoids are used in the clinic as anticoagulants. A specific pentasaccharide in heparinoids activates antithrombin III, resulting in inactivation of factor Xa and-when additional saccharides are present-inactivation of factor IIa. Structural and functional analysis of the heterogeneous heparinoids generally requires advanced equipment, is time consuming, and needs (extensive) sample preparation. In this study, a novel and fast method for the characterization of heparinoids is introduced based on reactivity with nine unique anti-heparin antibodies. Eight heparinoids were biochemically analyzed by electrophoresis and their reactivity with domain-specific anti-heparin antibodies was established by ELISA. Each heparinoid displayed a distinct immunoprofile matching its structural characteristics. The immunoprofile could also be linked to biological characteristics, such as the anti-Xa/anti-IIa ratio, which was reflected by reactivity of the heparinoids with antibodies HS4C3 (indicative for 3-O-sulfates) and HS4E4 (indicative for domains allowing anti-factor IIa activity). In addition, the immunoprofile could be indicative for heparinoid-induced side effects, such as heparin-induced thrombocytopenia, as illustrated by reactivity with antibody NS4F5, which defines a very high sulfated domain. In conclusion, immunoprofiling provides a novel, fast, and simple methodology for the characterization of heparinoids, and allows high-throughput screening of (new) heparinoids for defined structural and biological characteristics. PMID- 17909968 TI - Treatment retention in a prison-based residential sex offender treatment program. AB - This study assessed the role of static factors, a dynamic factor (motivation to change sexually deviant behavior), and an administrative factor in predicting treatment retention within a prison-based sex offender treatment program. The analyses also included assessing differences in initial levels of motivation and differences in beginning-versus end-of-treatment motivation scores for various types of program discharges. The sample consisted of 251 individuals who were admitted to a residential prison-based sex offender treatment program where 46% completed the program. Paired comparison t-tests showed higher motivation scores at the end of treatment only among treatment completers. Multivariate analyses showed that treatment retention was associated with higher initial motivation scores, higher levels of education and admission to treatment within 3 months of initial commitment to prison. Implications for motivational enhancement programming as well as for changes in admission criteria are discussed. PMID- 17909967 TI - Adverse effects from environmental mercury loads on breeding common loons. AB - Anthropogenic inputs of mercury (Hg) into the environment have significantly increased in the past century. Concurrently, the availability of methylmercury (MeHg) in aquatic systems has increased to levels posing risks to ecological and human health. We use the common loon (Gavia immer) as an upper trophic level bioindicator of aquatic Hg toxicity in freshwater lakes. Multiple endpoints were selected to measure potential negative impacts from MeHg body burdens on behavior, physiology, survival and reproductive success. A robust spatio-temporal dataset was used that included nearly 5,500 loon Hg measurements over an 18-year period. We measured significant changes related to elevated MeHg body burdens, including aberrant incubation behavior, lethargy, and wing area asymmetry. Mercury body burdens in adult loons increased an average of 8.4% per year. Increasing Hg body burdens reduced the number of fledged chicks per territorial pair, with highest risk loons producing 41% fewer fledged young than our reference group. Our multiple endpoints establish adverse effect thresholds for adult loons at 3.0 ug/g (wet weight) in blood and 40.0 ug/g (fresh weight) in feathers. Mercury contamination in parts of Maine and New Hampshire is a driving stressor for creating breeding population sinks. Standardized monitoring programs are needed to determine if population sinks occur elsewhere and to track aquatic ecosystem responses to changes in Hg emissions and deposition. PMID- 17909969 TI - The cumulative and sublethal effects of turbulence on erythrocytes in a stirred tank model. AB - Mechanical forces generated by prosthetic heart devices (artificial valves, artificial hearts, ventricular assist devices) have been known to cause damage and destruction of erythrocytes. Turbulent flow within such devices generates shear stresses and can induce cell damage. Current models of cell damage rate utilize only the power input per unit mass as a modeling parameter. A stirred tank reactor provides for a more extensive characterization of turbulence through eddy scale calculations. Through a simplified model, turbulence can be characterized by evaluating the Kolmogorov microscale. Our analysis of erythrocyte rupture in a stirred tank reactor suggests that parameters such as eddy wavelength and eddy velocity may better characterize and model the turbulent damage. Further, hemolysis of red blood cells by turbulent effects has been shown to have a fixed rate for constant levels of power input. Damage inflicted on the remaining, intact erythrocytes (sublethal damage) was evaluated by exposure to turbulence followed by osmotic fragility (OF) testing. Logistic models were fit to the OF data indicating a significant osmotic sensitivity in the sublethal damaged population between control and turbulence-exposed cells (chi(2) test; p < 0.001). This susceptibility indicates a significant cell population more susceptible to destruction as a result of turbulent exposure. This work has therefore helped identify optimization parameters for evaluating cell damage potential when engineering cardiovascular prosthetic devices. PMID- 17909970 TI - Quantification of rigidity in Parkinson's disease. AB - In this paper, a new method for quantification of rigidity in elbow joint of Parkinsonian patients is introduced. One of the most known syndromes in Parkinson's disease (PD) is increased passive stiffness in muscles, which leads to rigidity in joints. Clinical evaluation of stiffness in wrist and/or elbow, commonly used by clinicians, is based on Unified Parkinson's Disease Rating System (UPDRS). Subjective nature of this method may influence the accuracy and precision of evaluations. Hence, introducing an objective standard method based on quantitative measurements may be helpful. A test rig was designed and fabricated to measure range of motion and viscous and elastic components of passive stiffness in elbow joint. Measurements were done for 41 patients and 11 controls. Measures were extracted using Matlab-R14 software and statistic analyses were done by Spss-13. Relation between each computed measure and the level of illness were analyzed. Results showed a better correlation between viscous component of stiffness and UPDRS score compared to the elastic component. Results of this research may help to introduce a standard objective method for evaluation of PD. PMID- 17909971 TI - Biological activities of extracts from sumac (Rhus spp.): a review. AB - Sumac is the common name for a genus (Rhus) that contains over 250 individual species of flowering plants in the family Anacardiaceae. These plants are found in temperate and tropical regions worldwide, often grow in areas of marginal agricultural capacity, and have a long history of use by indigenous people for medicinal and other uses. The research efforts on sumac extracts to date indicate a promising potential for this plant family to provide renewable bioproducts with the following reported desirable bioactivities: antifibrogenic, antifungal, antiinflammatory, antimalarial, antimicrobial, antimutagenic, antioxidant, antithrombin, antitumorigenic, antiviral, cytotoxic, hypoglycaemic, and leukopenic. As well, the bioactive components can be extracted from the plant material using environmentally benign solvents that allow for both food and industrial end-uses. The favorable worldwide distribution of sumac also suggests that desirable bioproducts may be obtained at the source, with minimal transportation requirements from the source through processing to the end consumer. However, previous work has focussed in just a few members of this large plant family. In addition, not all of the species studied to date have been fully characterized for potential bioactive components and bioactivities. Thus, there remains a significant research gap spanning the range from lead chemical discovery through process development and optimization in order to better understand the full potential of the Rhus genus as part of global green technology based on bioproducts and bioprocesses research programs. PMID- 17909972 TI - Lymphoepithelial cyst of the pancreas. PMID- 17909973 TI - Pathophysiology of biliary-type abdominal pain. PMID- 17909974 TI - Extramedullary plasmacytoma involving the abdominal vessels and pancreas. PMID- 17909975 TI - Enterolith formation in the roux limb hepaticojejunostomy. AB - Enterolith in the Roux limb of Roux-en-Y hepaticojejunostomy is rare. We report a case of a Roux loop enterolith presenting with recurrent cholangitis. Cholescintigraphy and magnetic resonance imaging aided in the preoperative diagnosis. Intraoperatively, a large enterolith was extracted distal to the biliodigestive anastomosis. A kink of the small bowel was also noted distal to the stone. The mechanism for enterolith formation in the Roux loop is discussed. PMID- 17909976 TI - Liver and pancreatic injury induced by antituberculous therapy. PMID- 17909977 TI - Hepatotoxicity related to paraquat and diquat absorption through intact skin. PMID- 17909978 TI - Using Bland-Altman to assess agreement between two medical devices--don't forget the confidence intervals! AB - The limits of agreement approach of Bland and Altman is by far the most popular method for investigating statistical agreement between two measurement devices. This work presents the dangers of relying exclusively on the limits of agreement alone and argues that authors should always provide confidence intervals to assess the variability in the estimated limits. PMID- 17909979 TI - Sensitivity of FCV to recombinant feline interferon (rFeIFN). AB - Feline calicivirus cause feline respiratory diseases, and inactivated and attenuated vaccines are available for its prevention. Moreover, the presence of vaccine breakdown strains (VBS) is problematic. In Japan, feline recombinant interferon (rFeIFN) has been used for its treatment. However the method of compare with each strains has not established. To examine the relationship between the breakdown vaccine strain and rFeIFN sensitivity, the sensitivity of 47 field isolates to rFeIFN was determined. The Log PDD(50) values were normally distributed within the range 1.1-3.7, with a mean value of 2.3 +/- 0.64. Since 68.3% of the PDD values fell in the range of the mean +/- standard deviation, the values in the range 1.7-2.9, the lower values, and the higher values were defined as representing moderate, low, and high sensitivity, respectively. Among the 15 vaccine breakdown strains, strain Fukuoka9 showed a low sensitivity, but strains ML89, T58, and N74 were highly sensitive, showing no association with vaccine breakdown. The amino acid sequence changes specific to the low rFeIFN-sensitive Fukuoka-9 strain were found, suggesting that these sites are involved in rFeIFN sensitivity. PMID- 17909980 TI - The ambiguous pulmonary venoatrial junction: a new perspective. AB - PURPOSE: The pulmonary venoatrial junction (PVAJ) has recently received attention due to the widespread use of catheter ablation for atrial fibrillation. However, the literature lacks a consensus in the definition of the PVAJ. We aim to review the inconsistent definitions for the PVAJ and related implications in imaging and catheter ablation for atrial fibrillation. RESULTS: The PVAJ as described by embryology, gross anatomy, histology and imaging is ambiguous, leading to disparities in its definition. Because of differing definitions of the PVAJ, there is a broad range in the prevalence of anatomic variations, including (1) percentage of common pulmonary veins (10-79% on the left), (2) supernumerary pulmonary veins (10-42%) and (3) ostial diameter and shape. We postulate several reasons for this broad range in the described prevalence of anatomic variation of the PV as follows: (1) different definitions of the PVAJ, (2) different vantage points, (3) different imaging modalities, and (4) different prevalence of anatomic variants among different study populations. CONCLUSIONS: The ambiguous PVAJ with its gradual transition from the left atrium to the pulmonary veins defies precise definition even though it plays an important role in the management of atrial fibrillation. Physicians should be aware of variability in the language used to describe the PVAJ and resultant discrepancy in reported anatomical information. PMID- 17909981 TI - The quantification of dipyridamole induced changes in regional deformation in normal, stunned or infarcted myocardium as measured by strain and strain rate: an experimental study. AB - Strain rate imaging (SRI) during dobutamine stress-echocardiography (DSE) has been shown to differentiate between ischemic substrates based on the segmental response. Dipyridamole stress echo (DIPSE) is currently used as an alternative to DSE in detecting coronary artery disease. The aim of this study was: (a) to determine the normal response in peak-systolic myocardial strain (S) and strain rate (SR) during DIPSE and (b) to compare the S and SR responses of DSE and DIPSE in the same chronically ischemic/infarcted segments in the setting of single vessel disease. METHODS: The deformation response to DIPSE was studied in 7 normal pigs and in an additional 18 pigs, with a spectrum of ischemic substrates. S and SR data were extracted from a posterior wall "at risk" segment at baseline and during both DSE and DIPSE. The animals were divided into different ischemic substrate (stunning, non-transmural and transmural infarction), based on the DSE response as previously suggested. RESULTS: In normal myocardium, dipyridamole induced no changes in regional systolic deformation neither during nor after the infusion. Furthermore there was no detectable response in S and SR in segments with either a non-transmural or a transmural infarction. However, in myocardial segments with a DSE "stunning response", both end systolic S and peak-systolic SR tended to "normalize" at peak dipyridamole dose. CONCLUSIONS: These results suggest that dipyridamole does not induce changes in regional deformation in normal or (partially) infarcted myocardium. Only in stunned myocardium (in the setting of single-vessel disease), dipyridamole tends to normalize deformation. PMID- 17909982 TI - One stop cardiac investigation 'CT or echocardiography': beyond ejection fraction. PMID- 17909983 TI - Inhibition on Candida albicans biofilm formation using divalent cation chelators (EDTA). AB - Candida albicans can readily form biofilms on both inanimate and biological surfaces. In this study we investigated a means of inhibiting biofilm formation using EDTA (Ethylenediaminetetra-acetic acid), a divalent cation chelating agent, which has been shown to affect C. albicans filamentation. Candida albicans biofilms were formed in 96-well microtitre plates. Cells were allowed to adhere for 1, 2, and 4 h at 37 degrees C, washed in PBS, and then treated with different concentrations of EDTA (0, 2.5, 25, and 250 mM). EDTA was also added to the standardized suspension prior to adding to the microtiter plate and to a preformed 24 h biofilm. All plates were then incubated at 37 degrees C for an additional 24 h to allow for biofilm formation. The extent and characteristics of biofilm formation were then microscopically assessed and with a semi-quantitative colorimetric technique based on the use of an XTT-reduction assay. Northern blot analysis of the hyphal wall protein (HWP1) expression was also monitored in planktonic and biofilm cells treated with EDTA. Microscopic analysis and colorimetric readings revealed that filamentation and biofilm formation were inhibited by EDTA in a concentration dependent manner. However, preformed biofilms were minimally affected by EDTA (maximum of 31% reduction at 250 mM). The HWP1 gene expression was reduced in EDTA-treated planktonic and biofilm samples. These results indicate that EDTA inhibits C. albicans biofilm formation are most likely through its inhibitory effect on filamentation and indicates the potential therapeutic effects of EDTA. This compound may serve a non-toxic means of preventing biofilm formation on infections with a C. albicans biofilm etiology. PMID- 17909984 TI - Clinicians' evaluation of clinical ethics consultations in Norway: a qualitative study. AB - Clinical ethics committees have existed in Norway since 1996. By now all hospital trusts have one. An evaluation of these committees' work was started in 2004. This paper presents results from an interview study of eight clinicians who evaluated six committees' deliberations on 10 clinical cases. The study indicates that the clinicians found the clinical ethics consultations useful and worth while doing. However, a systematic approach to case consultations is vital. Procedures and mandate of the committees should be known to clinicians in advance to ensure that they know what to expect. Equally important is bringing all relevant facts, medical as well as psychosocial, into the discussion. A written report from the deliberation is also important for the committees to be taken seriously by the clinicians. This study indicates that the clinicians want to be included in the deliberation, and not only in the preparation or follow-up. Obstacles for referring a case to the committee are the medical culture's conflict aversion and its anxiety of being judged by outsiders. The committees were described as a court by some of the clinicians. This is a challenge for the committees in their attempt to balance support and critique in their consultation services. PMID- 17909985 TI - Survey on the experience in ethical decision-making and attitude of Pleven University hospital physicians towards ethics consultation. AB - BACKGROUND: Contemporary medical practice is complicated by many dilemmas requiring ethical sensitivity and moral reasoning. OBJECTIVE: To investigate physicians' experience in ethical decision-making and their attitude towards ethics consultation. METHODS: In a cross-sectional survey 126 physicians representing the main clinics of Pleven University hospital were investigated by a self-administered questionnaire. The following variables were measured: occurrence, nature and ways of resolving ethical problems; physicians' attitudes towards ethics consultation; physicians' opinions on qualities and skills of an ethics consultant, and socio-demographic characteristics. Data analysis included descriptive statistics, chi(2 )and t-test. RESULTS: Response rate was 88.9% (n = 112). Men and women were equally represented (48.2%-51.8%). The sample consisted of experienced physicians: 42.9% had 11-20 years experience, and 33% had 21-30 years. According to 84.8% of respondents, ethical problems have been discussed in their specialty. Predominant dilemmas included relationships with patients and relatives (76.8%) and team work (67.6%). Over (3/4) of physicians needed an advice in solving ethical problems. Ninety six percent responded positively to ethics consultation. They would mainly request it for resolving conflicts (72.5%), and in case of concern for the rightness of their decisions (52.7%). The image of an ethics consultant was built of clinical competence (70.9%), ability to deal with conflicts (59.1%), communication skills (58.2%), tolerance for different views (55.4%), and a special qualification in ethics (52.7%). CONCLUSIONS: The study underlined that Pleven University hospital physicians face similar ethical dilemmas as their colleagues in other countries do. The expressed positive attitudes to ethics consultation should serve as a basis for further research and development of ethics consultation services. PMID- 17909986 TI - A two-dimensional theory of health. AB - The starting point for the contemporary debate about theories of health should be the holistic theory of Lennart Nordenfelt, claims George Khushf, not the refuted theory of Christopher Boorse. The present paper is an attempt to challenge Nordenfelt and to present an alternative theory to his and other theories, including Boorse's. The main problems with Nordenfelt's theory are that it is relativistic, that it leads to counter-intuitive results as to what goals can count as healthy, that it focuses on the wrong kind of abilities, that it makes measuring health extra difficult, and that it does not give us a sufficient account of health, at most a necessary one. The alternative theory proposed is two-dimensional. First, health is to have developed the abilities and dispositions that members of one's culture typically develop, and be able to use them, in acceptable circumstances; and second, health is to experience positive moods and sensations, the kinds that have internal causes. The theory solves the problems attached to Nordenfelt's theory by not being individual relativistic, by eliminating the goals in the definition, by giving an alternative interpretation of "ability," by making health easier to measure, and by adding the dimension of well-being that, together with health as ability, not only gives us a necessary, but also a sufficient, account of health. PMID- 17909988 TI - True to oneself? Broad and narrow ideas on authenticity in the enhancement debate. AB - Our knowledge of the human brain and the influence of pharmacological substances on human mental functioning is expanding. This creates new possibilities to enhance personality and character traits. Psychopharmacological enhancers, as well as other enhancement technologies, raise moral questions concerning the boundary between clinical therapy and enhancement, risks and safety, coercion and justice. Other moral questions include the meaning and value of identity and authenticity, the role of happiness for a good life, or the perceived threats to humanity. Identity and authenticity are central in the debate on psychopharmacological enhancers. In this paper, I first describe the concerns at issue here as extensively propounded by Carl Elliott. Next, I address David DeGrazia's theory, which holds that there are no fundamental identity-related and authenticity-related arguments against enhancement technologies. I argue, however, that DeGrazia's line of reasoning does not succeed in settling these concerns. His conception of identity does not seem able to account for the importance we attach to personal identity in cases where personal identity is changed through enhancement technology. Moreover, his conception of authenticity does not explain the reason why we find inauthentic values objectionable. A broader approach to authenticity can make sense of concerns about changes in personal identity by means of enhancement technologies. PMID- 17909989 TI - Authentication of medicinal herbs using PCR-amplified ITS2 with specific primers. AB - Different parts of medicinal herbs have long been used as traditional Chinese drugs for treating many diseases, whereas materials of similar morphology and chemical fingerprints are often misidentified. Analyses of sequence variations in the nuclear ribosomal DNA (rDNA) internal transcribed spacer (ITS) have become a valid method for authentication of medicinal herbs at the intergenic and interspecific levels. DNA extracted from processed materials is usually severely degraded or contaminated by microorganisms, thus generates no or unexpected PCR products. The goal of this study is to apply the ITS fragments selectively amplified with two designed primer sets for efficient and precise authentication of medicinal herbs. The designed primers led to an accurate PCR product of the specific region in ITS2, which was confirmed with DNA extracted from 55 processed medicinal herbs belonging to 48 families. Moreover, the selectively amplified ITS2 authenticated five sets of easily confusable Chinese herbal materials. The designed primers were proven to be suitable for a broad application in the authentication of herbal materials. PMID- 17909990 TI - The evidence for abandoning the amniotic fluid index in favor of the single deepest pocket. AB - This study assessed whether the amniotic fluid index (AFI) or the single deepest pocket (SDP) is the best technique to estimate amniotic fluid volume. The AFI and SDP were compared to a dye-determined or directly measured amniotic fluid volume. A PUBMED search from 1990 to 2006 was conducted using the search terms "single deepest pocket" or "largest vertical pocket" or "maximum vertical pocket" or "2X1 pocket" AND "amniotic fluid index". One study compared the AFI and SDP to a dye determined amniotic fluid volume. There were 1219 publications that used the search term SDP-LVP-MVP versus 4378 using AFI. Twenty publications contained both the AFI and SDP, but only six compared the AFI and SDP. Both the AFI and the SDP poorly identified abnormal amniotic fluid volumes, and neither technique was superior to the other. The AFI identifies a significantly greater number of women as having oligohydramnios versus the SDP but without any difference in perinatal outcomes. Compared with SDP, AFI excessively characterizes a greater number of pregnancies as having oligohydramnios leading to more interventions without improvement in perinatal outcome. The AFI should be abandoned and the SDP used to estimate amniotic fluid volume. PMID- 17909991 TI - Neonatal macrocephaly: cerebral primitive neuroectodermal tumor or neuroblastoma as an infrequent cause--a case report and review of the literature. AB - We report a male term newborn presenting with a congenital macrocephaly 3.5 standard deviations above the median, with a wide and tense anterior fontanel, splayed calvarial sutures, and muscular hypotonia. Antenatal head circumferences were repeatedly below the median. A postnatal head ultrasound showed a large right intracerebral mass with right lateral ventricle compression, right temporal horn dilation, and right frontal horn enlargement with lateral displacement. Additional imaging by computed tomography scan and magnetic resonance imaging was performed. A decompression was performed and histology, immunohistochemistry, and molecular biology supported the diagnosis of a primitive neuroectodermal tumor. A MYCN gene amplification assay remained negative. The incidence of neonatal brain tumors is between 1.4 and 4.1/100,000 live births. Their most common presentation is macrocephaly, hydrocephalus, stillbirth, or diagnosis by pre- or postnatal imaging. Although hydrocephaly and intra- or extracranial hemorrhage are the most frequent causes of congenital macrocephaly, this should be initially investigated by head ultrasound. A suspected malignancy will be confirmed by histopathology, immunohistochemistry, and molecular biology. PMID- 17909992 TI - Estimation of glomerular filtration rate in preeclamptic patients. AB - Accurate estimation of the glomerular filtration rate (GFR) in patients with preeclampsia is often difficult or impossible to accomplish. In this study, the Cockcroft-Gault (CG), Modification of Diet in Renal Disease (MDRD), and MDRD2 formulas were evaluated for their accuracy in determining GFR in the setting of preeclampsia. The estimated GFR calculated from these formulas was compared with the creatinine clearance values obtained from a 24-hour urine collection in 209 preeclamptic patients recruited from five large hospitals. Additionally, a set of new equations that more accurately estimate GFR in preeclamptic patients based on ethnicity, preeclampsia GFR (PGFR), was created. Both the CG and MDRD formulas were inaccurate in predicting GFR in preeclamptic patients, and both were significantly less accurate than PGFR. In conclusion, current GFR estimation equations based on serum creatinine values in nonpregnant patients are not reliable measures of renal function in patients with preeclampsia. The use of a new (PGFR) formula is recommended. PMID- 17909993 TI - New directions for studying selection in nature: studies of performance and communities. AB - Natural and sexual selection are crucial factors in the evolutionary process, yet recent reviews show that researchers have focused narrowly on this topic, with the majority of research centered on the morphological traits of single species. However, in the past several years, several bodies of work have emerged that have examined both selection on performance capacity and selection in a community context, and our goal is to highlight these two growing areas and point toward future directions. Recent studies of selection on performance capacity point toward directional selection favoring high levels of performance, and we detected less evidence for selection favoring intermediate (i.e., stabilizing) or bimodal (i.e., disruptive) kinds of performance levels. Studies of selection in a community context, using the paradigm of indirect genetic effects, show significant community heritability and strong capacity for evolution to occur in a community context via the force of natural selection. For future directions, we argue that researchers should shift toward longer-term studies of selection on both individual species and communities, and we also encourage researchers to publish negative selection results for both performance and community studies to act as balancing influences on published positive selection results. PMID- 17909994 TI - Leptin levels and body composition of mice selectively bred for high voluntary locomotor activity. AB - Selective breeding produced four replicate lines of high-runner (HR) mice that run on wheels for approximately 2.7 times more revolutions per day than four unselected control lines. Previous studies found that HR mice of both sexes have lower body fat (isotope dilution at 15 wk of age) and that males (females not studied) have smaller retroperitoneal fat pads (17 wk). HR mice also exhibit elevated plasma corticosterone and insulin-stimulated glucose uptake by some hindlimb muscles but apparently do not differ in circulating insulin or glucose levels (males at 18 wk). Given their lower body fat and higher activity levels, we hypothesized that HR mice would have lower circulating leptin levels than controls. Female mice were given wheel access for 6 d at 7 wk of age, as part of the routine wheel testing for the selective breeding protocol, and then were killed after one additional week without wheels to reduce possible acute effects of activity on leptin. As hypothesized, serum leptin levels were significantly lower in HR mice. ANCOVA indicated that leptin was strongly positively correlated with both total body fat (measured by ether extraction) and body mass change from weaning, but HR mice still had significantly lower adjusted leptin levels (ANCOVA). Within HR lines but not within control lines, individual variation in leptin levels was negatively correlated with amount or speed of wheel running measured a week before being killed. Growth from weaning to euthanasia and body dry mass were lower in HR mice than in controls, but absolute dry masses of the ventricles, liver, gut, and uterus plus ovaries did not significantly differ, nor did percentage of the total dry mass as fat. HR mice offer a novel model for studying the causes and consequences of physiologically relevant variations in serum leptin. PMID- 17909995 TI - Physiological constraints and latitudinal breeding season in the Canidae. AB - Physiological strategies that maximize reproductive success may be phylogenetically constrained or might have a plastic response to different environmental conditions. Among mammals, Canidae lend themselves to the study of these two influences on reproductive physiology because all the species studied to date have been characterized as monestrous (i.e., a single ovulatory event per breeding season), suggesting a phylogenetic effect. Greater flexibility could be associated with environments that are less seasonal, such as the tropics; however, little is known for many of the species from this region. To compensate for this lack of data, two regressions were done on the length of the reproductive season relative to the latitudinal distribution of a species: one with raw data and another with phylogenetically independent contrasts. There was a significant negative relationship, independent of phylogeny, with canids that have longer breeding seasons occurring at lower latitudes. In contrast, the pervasiveness of monestrus within Canidae appears to be phylogenetically constrained by their pairing/packing life and is most likely associated with monogamy. The persistence of the monestrous condition is supported by a captive study where a tropical canid, the fennec fox, Vulpes zerda, never exhibited polyestrous cycles despite a constant photoperiod (12L : 12D). PMID- 17909996 TI - Hormones in the field: evolutionary endocrinology of juvenile hormone and ecdysteroids in field populations of the wing-dimorphic cricket Gryllus firmus. AB - Virtually no published information exists on insect endocrine traits in natural populations, which limits our understanding of endocrine microevolution. We characterized the hemolymph titers of juvenile hormone (JH) and ecdysteroids (ECDs), two key insect hormones, in field-collected short-winged, flightless (SW) and long-winged, flight-capable (LW(f)) morphs of the cricket Gryllus firmus. The JH titer exhibited a dramatic circadian rhythm in the LW(f) morph but was temporally constant in the flightless SW morph. This pattern was consistent in each of three years; in young, middle-aged, and older G. firmus; and in three other cricket species. The ECD titer was considerably higher in SW than in LW(f) females but did not exhibit temporal variation in any morph and did not differ between male morphs. JH and ECD may control different aspects of the morph specific trade-off between nocturnal dispersal and reproduction. Results confirm and extend laboratory studies on young female G. firmus; most, but not all, important aspects of morph-specific differences in JH and ECD titers can be extrapolated from field to laboratory environments and vice versa. Hormone titers in Gryllus are more complex than those proposed in evolutionary endocrine models. Directly measuring hormone titer variation remains a fundamentally important task of insect evolutionary endocrinology. PMID- 17909997 TI - Growth, development, and nutritional physiology of grasshoppers from subarctic and temperate regions. AB - Despite the importance of developmental rate, growth rate, and size at maturity in the life history of poikliotherms, the trade-offs among these traits and selection pressures involved in the evolution of these traits are not well understood. This study compared these traits in a grasshopper, Melanoplus sanguinipes F. (Orthoptera: Acrididae), from two contrasting geographical regions, subarctic Alaska and temperate Idaho. The growing season in the interior of Alaska is about 80 d shorter than at low-elevation sites in Idaho. We hypothesized that the Alaskan grasshoppers would show more rapid growth and development than grasshoppers from Idaho, at the cost of greater sensitivity to food quality. On a diet of lettuce and wheat bran, grasshoppers from Alaska developed from egg hatch to adult more rapidly than those from Idaho at each of three different temperature regimes. Averaged over all temperature treatments, the weight of the Alaskan grasshoppers was about 5% less than that of the Idaho grasshoppers at the adult molt. Feeding and digestive efficiencies were determined for the final two instars using two meridic diets: one with a high concentration of nutrients and the other with the same formulation but diluted with cellulose. Alaskan grasshoppers again developed more rapidly, weighed less, and had faster growth rates than those from Idaho. Alaskan grasshoppers supported their more rapid growth by increasing postingestive efficiencies; that is, they had higher conversion rates of digested matter to biomass on the high-quality diet, greater assimilation of food on the low-quality diet, and greater efficiency of nitrogen assimilation or retention on both diets. There was no evidence that performance of Alaskan grasshoppers suffered any more than that of the Idaho grasshoppers on the low-quality diet. PMID- 17909998 TI - The relationships among sprint performance, voluntary swimming activity, and social dominance in juvenile rainbow trout. AB - The specific objectives of this study were to determine whether sprint performance in juvenile rainbow trout is correlated with either voluntary swimming activity or aggressive behaviors and to determine the reciprocal: the effect of swimming activity and aggression on sprint performance. Sprint performance was assessed by rapidly accelerating trout (5-7-cm fork length) to a fixed velocity (40, 42, or 45 cm s(-1)) and then holding them at that velocity until fatigue. There was considerable interindividual variation in sprint performance not explained by variations in body size, but intraindividual performance was highly repeatable over at least 2 mo. Voluntary swimming was measured as the frequency of transits (voluntary transit activity, VTA) between two identical tanks via a connecting channel with two different flow regimes: zero or minimum velocity (0 or 2.5 cm s(-1)) and high velocity (84 cm s(-1)). There was a strong correlation between sprint performance and VTA in minimal current but no correlation in high current. Furthermore, sprint performance did not predict the outcome of dominance encounters. Experience with rapid acceleration, especially when voluntary, led to a pronounced improvement in sprint performance in proportion to the number of acceleration events. Social dominance encounters had a more complex effect: a significant reduction in sprint performance in previously high-performance sprinters and the reverse for low performers. We propose that there are four independent axes of interindividual variation in juvenile rainbow trout: spontaneous and rheotaxis-stimulated locomotor activity, aggressive activity, and the trainability of sprint performance. The independence of these axes has the potential to produce a much larger diversity in behavioral and ultimately physiological phenotypes than would be produced if the axes were linked. PMID- 17909999 TI - High rates of energy expenditure and water flux in free-ranging Point Reyes mountain beavers Aplodontia rufa phaea. AB - We measured water flux and energy expenditure in free-ranging Point Reyes mountain beavers Aplodontia rufa phaea by using the doubly labeled water method. Previous laboratory investigations have suggested weak urinary concentrating ability, high rates of water flux, and low basal metabolic rates in this species. However, free-ranging measurements from hygric mammals are rare, and it is not known how these features interact in the environment. Rates of water flux (210+/ 32 mL d(-1)) and field metabolic rates (1,488+/-486 kJ d(-1)) were 159% and 265%, respectively, of values predicted by allometric equations for similar-sized herbivores. Mountain beavers can likely meet their water needs through metabolic water production and preformed water in food and thus remain in water balance without access to free water. Arginine-vasopressin levels were strongly correlated with rates of water flux and plasma urea : creatinine ratios, suggesting an important role for this hormone in regulating urinary water loss in mountain beavers. High field metabolic rates may result from cool burrow temperatures that are well below lower critical temperatures measured in previous laboratory studies and suggest that thermoregulation costs may strongly influence field energetics and water flux in semifossorial mammals. PMID- 17910000 TI - An energy-based body temperature threshold between torpor and normothermia for small mammals. AB - Field studies of use of torpor by heterothermic endotherms suffer from the lack of a standardized threshold differentiating torpid body temperatures (T(b)) from normothermic T(b)'s. This threshold can be more readily observed if metabolic rate (MR) is measured in the laboratory. I digitized figures from the literature that depicted simultaneous traces of MR and T(b) from 32 respirometry runs for 14 mammal species. For each graph, I quantified the T(b) measured when MR first began to drop at the onset of torpor (T(b-onset)). I used a general linear model to quantify the effect of ambient temperature (T(a)) and body mass (BM) on T(b onset). For species lighter than 70 g, the model was highly significant and was described by the equation Tb-onset=(0.055+/-0.014)BM+(0.071+/-0.031)Ta+(31.823+/ 0.740). To be conservative, I recommend use of these model parameters minus 1 standard error, which modifies the equation to Tb-onset-1 SE=(0.041)BM+(0.040)Ta+31.083. This approach provides a standardized threshold for differentiating torpor from normothermia that is based on use of energy, the actual currency of interest for studies of torpor in the wild. Few laboratory studies have presented the time-course data required to quantify T(b-onset), so more data are needed to validate this relationship. PMID- 17910001 TI - Deer mouse aerobic performance across altitudes: effects of developmental history and temperature acclimation. AB - Aerobic physiology at high altitudes has been studied in many animals. Prior work on laboratory-bred deer mice (a species with a wide altitudinal range) showed depression of aerobic capacity at high altitude, even after acclimation. However, wild deer mice show no reduction in thermogenic performance at high altitude, and performance limits seem to be due to physiological and anatomical adjustments to environmental temperature and not to oxygen availability. We asked whether across altitude performance differences exist in deer mice after accounting for temperature acclimation (approximately 5 degrees and 20 degrees -25 degrees C) and prenatal and neonatal development altitude (340 vs. 3,800 m). We measured maximal thermogenic oxygen consumption (VO2sum) in cold exposure and ran mice on a treadmill to elicit maximal exercise oxygen consumption (VO2max). We found a 10% reduction in VO2max at 3,800 m compared with that at 340 m; thus, the mice were able to compensate for most of the 37% reduction in oxygen availability at the higher altitude. Development altitude did not affect VO2max. There was no effect of test altitude or development altitude on VO2sum in warm-acclimated animals, but both test and development altitude strongly affected VO2sum in cold acclimated mice, and compensation for hypoxia at 3,800 m was considerably less than that for exercise. PMID- 17910003 TI - Chirality and chemical processes: a few afterthoughts. AB - Chirality and chiral have become terms that pervade a wide range of disciplines in physical and life sciences. Although such terms are precisely defined, their use often engenders confusion and ambiguity. Perhaps, the most improper use of chirality, yet widely accepted, is related to its association with stereodynamics and physico-chemical transformations, such as chiral discrimination, chiral resolution, chiral recognition, chiral synthesis, and so on. Even though this conceptual perversion has been highlighted by renowned stereochemists, it has become a recurring keyword and a hot message in modern literature. It is timely to renew the correct use and context in forums such as the present journal, adding further reflections that may help both beginners and practitioners. This short article is not intended to criticize or highlight errors, but rather to encourage a level of rigor and the use of statements, which should be universally correct. PMID- 17910002 TI - Carotenoid coloration in greenfinches is individually consistent irrespective of foraging ability. AB - Carotenoid-based plumage coloration of birds has been hypothesized to honestly reflect individual quality, either because carotenoids are difficult to acquire via food or because of a trade-off in allocation of carotenoids between maintenance and signaling functions. We tested whether differential foraging ability is a necessary precondition for maintaining individual differences in carotenoid-based plumage coloration in male greenfinches (Carduelis chloris). Wild-caught birds were brought into captivity, where half of them were supplemented with carotenoids while the other half was maintained on a carotenoid poor diet. Color of the yellow parts of tail feathers, grown under natural conditions, was compared with that of the replacement feathers, grown in captivity. Carotenoid supplementation increased feather chroma (saturation). Color of wild-grown feathers significantly correlated with the color of lab-grown feathers. This result demonstrates the existence of a significant component of variation in carotenoid coloration, which reflects physiological qualities or genetic differences among individuals independent of foraging ability. Among both experimental groups, plasma carotenoid concentration during feather growth strongly correlated with chroma of the feathers grown in captivity. This indicates that carotenoid-based plumage coloration can reveal circulating carotenoid levels over a very wide range of concentrations, suggesting the ample signaling potential of such a mechanism. PMID- 17910004 TI - New Tabu Search based global optimization methods outline of algorithms and study of efficiency. AB - The study presents two new nonlinear global optimization routines; the Gradient Only Tabu Search (GOTS) and the Tabu Search with Powell's Algorithm (TSPA). They are based on the Tabu-Search strategy, which tries to determine the global minimum of a function by the steepest descent-mildest ascent strategy. The new algorithms are explained and their efficiency is compared with other approaches by determining the global minima of various well-known test functions with varying dimensionality. These tests show that for most tests the GOTS possesses a much faster convergence than global optimizer taken from the literature. The efficiency of the TSPA compares to the efficiency of genetic algorithms. PMID- 17910006 TI - Structure and dynamics of phospholipid bilayers using recently developed general all-atom force fields. AB - Two fully hydrated pure-species phospholipids bilayers, 1,2-dimyristoyl-sn glycero-3-phosphocholine (DMPC) and 1,2-dioleoyl-sn-glycero-3-phosphorylcholine (DOPC), in the fluid phase and explicit solvent have been studied using molecular dynamics simulation. Atom interactions were modeled using recently developed force fields based on AMBER with full atomistic details. Several representative liquid phase properties for the structure and dynamics of lipids with different length of hydrocarbon chains and different level of saturation have been reproduced without artificially biasing the system in order to match experimental data. In particular, as the new GAFF (General Amber Force Field) has not been explicitly developed to reproduce lipid characteristics and is naturally compatible with standard AMBER nucleic acids and proteins parameters, it is here proven a promising tool to study mixed lipid-protein processes as protein activity dependence on membrane composition, permeation of solute across membranes, and other cellular processes. PMID- 17910007 TI - The sources and manifestations of stress amongst school-aged dyslexics, compared with sibling controls. AB - All school children experience stress at some point in their school careers. This study investigates whether dyslexic children, by way of their educational and social difficulties, experience higher levels of stress at school. The School Situation Survey was used to investigate both the sources and manifestations of stress amongst dyslexic children and non-dyslexic sibling controls. Samples were broken down by gender, age and the size of families. Results suggest significant differences between the groups, with dyslexics in academic years 3-5 experiencing the highest stress levels, specifically in interactions with teachers, worries over academic examinations (SATs) and performance testing, causing emotional (fear, shyness and loneliness) and physiological (nausea, tremors or rapid heart beat) manifestations. Results also suggest that dyslexics in larger families (3-4 sibling families) experience greater stress in interactions with their peers, than those in smaller families (two sibling families)--possibly from unfair sibling comparison. PMID- 17910008 TI - The use of Gaussian quadrature for estimation in frailty proportional hazards models. AB - In this paper, we propose a novel Gaussian quadrature estimation method in various frailty proportional hazards models. We approximate the unspecified baseline hazard by a piecewise constant one, resulting in a parametric model that can be fitted conveniently by Gaussian quadrature tools in standard software such as SAS Proc NLMIXED. We first apply our method to simple frailty models for correlated survival data (e.g. recurrent or clustered failure times), then to joint frailty models for correlated failure times with informative dropout or a dependent terminal event such as death. Simulation studies show that our method compares favorably with the well-received penalized partial likelihood method and the Monte Carlo EM (MCEM) method, for both normal and Gamma frailty models. We apply our method to three real data examples: (1) the time to blindness of both eyes in a diabetic retinopathy study, (2) the joint analysis of recurrent opportunistic diseases in the presence of death for HIV-infected patients, and (3) the joint modeling of local, distant tumor recurrences and patients survival in a soft tissue sarcoma study. The proposed method greatly simplifies the implementation of the (joint) frailty models and makes them much more accessible to general statistical practitioners. PMID- 17910009 TI - Baseline and treatment effect heterogeneity for survival times between centers using a random effects accelerated failure time model with flexible error distribution. AB - Nowadays, most clinical trials are conducted in different centers and even in different countries. In most multi-center studies, the primary analysis assumes that the treatment effect is constant over centers. However, it is also recommended to perform an exploratory analysis to highlight possible center by treatment interaction, especially when several countries are involved. We propose in this paper an exploratory Bayesian approach to quantify this interaction in the context of survival data. To this end we used and generalized a random effects accelerated failure time model. The generalization consists in using a penalized Gaussian mixture as an error distribution on top of multivariate random effects that are assumed to follow a normal distribution. For computational convenience, the computations are based on Markov chain Monte Carlo techniques. The proposed method is illustrated on the disease-free survival times of early breast cancer patients collected in the EORTC trial 10854. PMID- 17910010 TI - Comment on 'The partial area under the summary ROC curve'. PMID- 17910011 TI - Biologically stable 2-5A analogues containing 3'-O,4'-C-bridged adenosine as potent RNase L agonists. PMID- 17910013 TI - Microwave-assisted palladium-catalyzed direct arylation of 1,4-disubstituted 1,2,3-triazoles with aryl chlorides. AB - Treatment of 1,4-disubstituted 1,2,3-triazoles with aryl chlorides in the presence of potassium carbonate under palladium catalysis and microwave irradiation at 250 degrees C for 15 min leads to arylation of the triazole at the 5-position. A variety of functional groups, including ester and hydroxy groups, are compatible. The procedure is suitable for the regioselective preparation of trisubstituted triazoles. Microwave irradiation accelerates the reaction, thus allowing the rapid synthesis of trisubstituted triazoles, which are difficult to synthesize selectively. PMID- 17910012 TI - A novel method of cellular labeling: anchoring mr-imaging reporter particles on the outer cell surface. PMID- 17910014 TI - Intrinsic deuterium kinetic isotope effects in glutamate mutase measured by an intramolecular competition experiment. PMID- 17910015 TI - Buffering dissociation/formation reaction of biogenic calcium carbonate. AB - The oscillating stability of coral reef seawater pH has been maintained at around physiological pH values over the past 300 years (Pelejero et al., 2005). The stability mechanism of its pH has been interpreted in terms of the buffering dissolution/formation reaction of CaCO(3) as well as the proton consumption/generation reaction in CaCO(3)-saturated water. Here the pH-dependent solubility product [HCO(3)(-)][Ca(2+)] has been derived on the basis of the actual pH-dependent reactions for the atmospheric CO(2)/CO(2 (aq.))/HCO(3)( )/CO(3)(2-)/Ca(2+)/CaCO(3) system. Overbasic pH peaks appeared between pH approximately 8 and approximately 9.5 during sodium hydroxide titration, as a result of simultaneous CaCO(3) formation and proton generation. The spontaneous and prompt water pH recovery from the acidic to the physiological range has been confirmed by the observation of acid/base time evolution, because of simultaneous CaCO(3) dissolution and proton consumption. The dissolution/formation of CaCO(3) in water at pH 7.5-9 does not take place without a proton consumption/generation reaction, or a buffering chemical reaction of HCO(3)(-)+Ca(2+)right arrow over left arrowCaCO(3)+H(+). SEM images of the CaCO(3) fragments showed that the acid water ate away at the CaCO(3) formed at physiological pH values. Natural coral reefs can thus recover the physiological pH levels of seawater from the acidic range through partial dissolution of their own skeletons. PMID- 17910016 TI - Hydrocarbon-soluble calcium hydride: a "worker-bee" in calcium chemistry. AB - The reactivity of the hydrocarbon-soluble calcium hydride complex [{CaH(dipp nacnac)(thf)}(2)] (1; dipp-nacnac=CH{(CMe)(2,6-iPr(2)C(6)H(3)N)}(2)) with a large variety of substrates has been investigated. Addition of 1 to C=O and C=N functionalities gave easy access to calcium alkoxide and amide complexes. Similarly, reduction of the C[triple chemical bond]N bond in a cyanide or an isocyanide resulted in the first calcium aldimide complexes [Ca{N=C(H)R}(dipp nacnac)] and [Ca{C(H)=NR}(dipp-nacnac)], respectively. Complexation of 1 with borane or alane Lewis acids gave the borates and alanates as contact ion pairs. In reaction with epoxides, nucleophilic ring-opening is observed as the major reaction. The high reactivity of hydrocarbon-soluble 1 with most functional groups contrasts strongly with that of insoluble CaH(2), which is essentially inert and is used as a common drying agent. Crystal structures of the following products are presented: [{Ca{OC(H)Ph(2)}(dipp-nacnac)}(2)], [{Ca{N=C(H)Ph}(dipp nacnac)}(2)], [{Ca{C(H)=NC(Me)(2)CH(2)C(Me)(3)}(dipp-nacnac)}(2)], [{Ca{C(H)=NCy}(dipp-nacnac)}(2)], [Ca(dipp-nacnac)(thf)](+)[H(2)BC(8)H(14)](-) and [{Ca(OCy)(dipp-nacnac)}(2)]. The generally smooth and clean conversions of 1 with a variety of substrates and the stability of most intermediates against ligand exchange make 1 a valuable key precursor in the syntheses of a wide variety of beta-diketiminate calcium complexes. PMID- 17910017 TI - Supramolecular helical columns from the self-assembly of chiral rods. AB - Chiral-bridged rod molecules (CBRs) that consisted of bis(penta-p-phenylene) conjugated to an opened or closed chiral bridging group as a rigid segment and oligoether dendrons as flexible segments were synthesized and characterized. In the bulk state, both molecules self-assemble into a hexagonal columnar structure, as confirmed by X-ray scatterings and transmission electron microscopy (TEM) observations. Interestingly, these structures display opposite Cotton effects in the chromophore of the aromatic unit in spite of the same chirality (R,R) of the chiral bridging groups. The molecules were observed to self-assemble into cylindrical micellar aggregates in aqueous solution, as confirmed by light scattering and TEM investigations, and exhibit intense signals in the circular dichroism (CD) spectra, which are indicative of one-handed helical conformations. The CD spectra of each molecule showed opposite signals to each other, which were similar to those in the bulk. Notably, when the opened CBR was added to a solution of closed CBRs up to a certain concentration, the CD signal of the closed CBR was amplified. This implies that both molecules co-assemble into a one handed helical structure because the opened chiral bridge is conformationally flexible, which is inverted to co-assemble with the closed CBR. These results demonstrate that small structural modifications of the chiral moiety can transfer the chiral information to a supramolecular assembly in the opposite way. PMID- 17910018 TI - Efficient and convenient heterogeneous palladium-catalyzed regioselective deuteration at the benzylic position. AB - The Pd/C-catalyzed efficient and regioselective hydrogen-deuterium (H-D) exchange reaction on the benzylic site proceeded in D2O in the presence of a small amount of H2 gas. The use of the Pd/C-ethylenediamine complex [Pd/C(en)] as a catalyst instead of Pd/C led to the efficient deuterium incorporation into the benzylic site of O-benzyl protective groups without hydrogenolysis. These H-D exchange reactions provide a post synthetic and D(2)-gas-free deuterium-labeling method on a wide variety of benzylic sites using D2O as the deuterium source and heterogeneous Pd/C or Pd/C(en) as a reusable heterogeneous palladium catalyst under mild and neutral conditions. PMID- 17910019 TI - Combining computational and biochemical studies for a rationale on the anti aromatase activity of natural polyphenols. AB - Aromatase, an enzyme of the cytochrome P450 family, is a very important pharmacological target, particularly for the treatment of breast cancer. The anti aromatase activity of a set of natural polyphenolic compounds was evaluated in vitro. Strong aromatase inhibitors including flavones, flavanones, resveratrol, and oleuropein, with activities comparable to that of the reference anti aromatase drug aminoglutethimide, were identified. Through the application of molecular modeling techniques based on grid-independent descriptors and molecular interaction fields, the major physicochemical features associated with inhibitory activity were disclosed, and a putative virtual active site of aromatase was proposed. Docking of the inhibitors into a 3D homology model structure of the enzyme defined a common binding mode for the small molecules under investigation. The good correlation between computational and biological results provides the first rationalization of the anti-aromatase activity of polyphenolic compounds. Moreover, the information generated in this approach should be further exploited for the design of new aromatase inhibitors. PMID- 17910021 TI - Infrared spectra of protonated uracil, thymine and cytosine. AB - The gas-phase structures of protonated uracil, thymine, and cytosine are probed by using mid-infrared multiple-photon dissociation (IRMPD) spectroscopy performed at the Free Electron Laser facility of the Centre Laser Infrarouge d'Orsay (CLIO), France. Experimental infrared (IR) spectra are recorded for ions that were generated by electrospray ionization, isolated, and then irradiated in a quadrupole ion trap; the results are compared to the calculated infrared absorption spectra of the different low-lying isomers (computed at the B3LYP/6 31++G(d,p) level). For each protonated base, the global energy minimum corresponds to an enolic tautomer, whose infrared absorption spectrum matched very well with the experimental IRMPD spectrum, with the exception of a very weak IRMPD signal observed at about 1800 cm(-1) in the case of the three protonated bases. This signal is likely to be the signature of the second-energy-lying oxo tautomer. We thus conclude that within our experimental conditions, two tautomeric ions are formed which coexist in the quadrupole ion trap. PMID- 17910020 TI - Patterns and predictors of medication compliance, diversion, and misuse in adult prescribed methylphenidate users. AB - OBJECTIVES: To examine patterns and predictors of medication compliance, diversion, and misuse in a sample of adults with prescriptions for the stimulant medication methylphenidate (MPH). METHODS: Sixty-six adults currently prescribed MPH (53% male) completed structured interviews and provided details regarding their medication and other substance use histories. RESULTS: On average, participants reported using their medication as prescribed on 14.5 (SD 11.7) of the past 30 days; 44% admitted to diverting it and 29% admitted to inappropriate use. While analyses revealed that medication misuse, diversion, and level of compliance were interrelated and all associated with concurrent illicit substance use, each also had other distinct associations. Specifically, MPH misuse was associated with the use of illicit stimulants such as amphetamine and cocaine, diversion with age and age of MPH prescription, and compliance with participation in an attention deficit hyperactivity disorder (ADHD) support group. Regression analyses revealed that misuse and poor compliance were both best predicted by concurrent illicit substance use, while the model that best predicted diversion included age of first MPH prescription (younger) and MPH misuse. CONCLUSIONS: Poor medication compliance, diversion, and misuse are relatively common and interrelated among adult MPH users. MPH prescriptions should be monitored closely in individuals with histories of illicit substance use. PMID- 17910023 TI - Distinction of ortho- and para-xylene by femtosecond-laser mass spectrometry. PMID- 17910024 TI - IR and NMR properties of ionic liquids: do they tell us the same thing? AB - We used a combination of theoretical and experimental methods to derive the spectroscopic properties of imidazolium-based ionic liquids. Vibrational frequencies, NMR chemical shifts, and quadrupole coupling constants react in comparable manner to changes in the chemical environment. This suggests that both the IR and the NMR spectroscopic properties reflect a similar type of electronic perturbation caused by hydrogen bonding. These relationships of the spectroscopic properties provide detailed information about structural complexes and may thus serve as good indicators of ion-pair formation. They also help to decide which spectroscopic tool is the most sensitive for investigating molecular interactions. The measurement of only one spectroscopic property allows the prediction of other properties that cannot be so easily measured. In some cases, this is the only way to obtain reliable coupling constants for deriving molecular correlation times from macroscopic NMR relaxation times, thus opening a new path for studying structure-dynamics relations in ionic liquids. PMID- 17910022 TI - Conformational influence on the type of stabilization of sulfur radical cations in cyclic peptides. AB - The free-radical chemistry of two oxidized cyclic dipeptides is investigated using time-resolved optical and conductivity detection. Two cyclic dipeptides, cyclo-Gly-L-Met and cyclo-D-Met-L-Met, are synthesized and irradiated with nanosecond pulses of electrons, which initiate the oxidation of the methionine side chains with hydroxyl radicals from the radiolysis of water. The cyclic peptides are taken to be models for the interior of proteins where there are no terminal groups. This opens up the possibility that neighboring-group effects can be studied directly between the initially formed sulfur radical cations and the heteroatoms associated with the peptide bonds. Such complexation of the sulfur radical cations is observed with the amide nitrogen atoms. In addition, intermolecular stabilization with the unoxidized sulfur atoms on separate cyclic dipeptide molecules is observed. Little or no intramolecular stabilization by the unoxidized sulfur in the neighboring methionine occurs in cyclo-D-Met-L-Met, in contrast to the previously observed intramolecular sulfur stabilization of the sulfur radical cation in the isomer cyclo-L-Met-L-Met. This contrasting behavior is rationalized by conformational differences in the two isomers as seen through molecular-modeling simulations. The implications for the oxidation of the protein calmodulin, which contains multiple residues of methionine, are discussed as having analogous determining factors. PMID- 17910025 TI - Strategies for measurements of pseudocontact shifts in protein NMR spectroscopy. AB - Paramagnetic metal ions bound to proteins generate a dipolar field that can be accurately probed by pseudocontact shifts (PCS) displayed by the protein's nuclear spins. PCS are highly useful for determining the coordinates of individual spins in the molecule and for rapid structure determinations of entire protein-protein and protein-ligand complexes. However, PCS measurements require reliable resonance assignments for the molecule in its paramagnetic state and in a diamagnetic reference state. This article discusses different approaches for pairwise resonance assignments, with emphasis on a strategy which exploits chemical exchange between the two states. PMID- 17910026 TI - Quantitative ATP synthesis in human liver measured by localized 31P spectroscopy using the magnetization transfer experiment. AB - The liver plays a central role in intermediate metabolism. Accumulation of liver fat (steatosis) predisposes to various liver diseases. Steatosis and abnormal muscle energy metabolism are found in insulin-resistant and type-2 diabetic states. To examine hepatic energy metabolism, we measured hepatocellular lipid content, using proton MRS, and rates of hepatic ATP synthesis in vivo, using the 31P magnetization transfer experiment. A suitable localization scheme was developed and applied to the measurements of longitudinal relaxation times (T1) in six healthy volunteers and the ATP-synthesis experiment in nine healthy volunteers. Liver 31P spectra were modelled and quantified successfully using a time domain fit and the AMARES (advanced method for accurate, robust and efficient spectral fitting of MRS data with use of prior knowledge) algorithm describing the essential components of the dataset. The measured T1 relaxation times are comparable to values reported previously at lower field strengths. All nine subjects in whom saturation transfer was measured had low hepatocellular lipid content (1.5 +/- 0.2% MR signal; mean +/- SEM). The exchange rate constant (k) obtained was 0.30 +/- 0.02 s(-1), and the rate of ATP synthesis was 29.5 +/- 1.8 mM/min. The measured rate of ATP synthesis is about three times higher than in human skeletal muscle and human visual cortex, but only about half of that measured in perfused rat liver. In conclusion, 31P MRS at 3 T provides sufficient sensitivity to detect magnetization transfer effects and can therefore be used to assess ATP synthesis in human liver. PMID- 17910027 TI - Control of beta globin genes. AB - The developmental changes in expression of the beta like genes from embryonic to adult stages of human life are controlled at least partially at the level of the promoter sequences of these genes and their binding factors, and competition for promoter specific interactions with the locus control region (LCR). In recent years, the control of beta globin genes has also been investigated at the level of chromatin structure involving the chemical modification of histones and their remodelling by DNA dependent ATPases (SMARCA) containing protein complexes. The role of intergenic RNA is also being investigated with renewed interest. Although a wealth of information on the structure/function relationship of the LCR and globin promoters has been gathered over more than two decades, the fundamental nature of the control of these genes at the molecular level is still not completely understood. In the following pages, we intend to briefly describe the progress made in the field and discuss future directions. PMID- 17910029 TI - Immunosuppressive drug-free operational immune tolerance in human kidney transplant recipients: Part I. Blood gene expression statistical analysis. AB - Survival of solid organ grafts depends on life-long immunosuppression, which results in increased rates of infection and malignancy. Induction of tolerance to allografts would represent the optimal solution for controlling both chronic rejection (CR) and side effects of immunosuppression. Although spontaneous "operational tolerance" can occur in human kidney transplantation, the lack of noninvasive peripheral blood biological markers of this rare phenomenon precludes the identification of potentially tolerant patients in whom immunosuppression could be tapered as well as the development of new tolerance inducing strategies. Here, the potential of high throughput microarray technology to decipher complex pathologies allowed us to study the peripheral blood specific gene expression profile and corresponding EASE molecular pathways associated to operational tolerance in a cohort of human kidney graft recipients. In comparison with patients with CR, tolerant patients displayed a set of 343 differentially expressed genes, mainly immune and defense genes, in their peripheral blood mononuclear cells (PBMC), of which 223 were also different from healthy volunteers. Using the expression pattern of these 343 genes, we were able to classify correctly >80% of the patients in a cross-validation experiment and classified correctly all of the samples over time. Collectively, this study identifies a unique PBMC gene signature associated with human operational tolerance in kidney transplantation by a classical statistical microarray analysis and, in the second part, by a nonstatistical analysis. PMID- 17910028 TI - Control of osteopontin signaling and function by post-translational phosphorylation and protein folding. AB - Osteopontin (OPN) plays roles in a variety of cellular processes from bone resorption and extracellular matrix (ECM) remodeling to immune cell activation and inhibition of apoptosis. Because it binds receptors (integrins, CD44 variants) typically engaged by ECM molecules, OPN acts as a "soluble" ECM molecule. A persistent theme throughout the characterization of how OPN functions has been the importance of phosphorylation. The source of the OPN used in specific experiments and the location of modified sites is an increasingly important consideration for OPN research. We review briefly some of the ways OPN impacts on the biology of mammalian systems with an emphasis on the importance of serine phosphorylation in modulating its signaling ability. We describe experiments that support the hypothesis that differences in the post translational phosphorylation of OPN expressed by different cell types regulate how it impacts on target cells. Analyses of OPN's potential secondary structure reveal a possible beta-sheet conformation that offers an interpretation of certain experimental observations, specifically the effect of thrombin cleavage; it is consistent with an interaction between the C-terminal region of the protein and the central integrin-binding RGD sequence. PMID- 17910030 TI - Dieting in Spanish adolescent girls. AB - This article examines the relation between dieting and non-dieting adolescent girls and body mass index (BMI), eating attitudes and the influence of the aesthetic body shape model. Three hundred forty-nine Spanish adolescent girls in their second year of secondary education were selected. The research design was a cross-sectional ex post facto study and validated measures were used to assess the main variables. Fourteen per cent of the sample was on a diet. Of this group, 69% were overweight, 70% had disordered eating attitudes and 70% were influenced by the current aesthetic body shape model. In all situations, the scores for dieters were significantly higher than those for non-dieters. Although there is not yet any clear consensus with regards to how adolescents interpret the term 'diet', the results indicate major differences between dieters and non-dieters and that those who self-report that they are on a diet present elements of the risk of developing eating and weight disorders. PMID- 17910031 TI - Fatal falls overboard on commercial fishing vessels in Alaska. AB - BACKGROUND: Falls overboard are a major contributor to commercial fishing fatalities in Alaska. The National Institute for Occupational Safety and Health has repeatedly identified falls overboard as a critical issue in commercial fishing safety. This article describes the problem of falls overboard and discusses possible ways to reduce the risk factors. METHODS: Data from the Alaska Occupational Injury Surveillance System on fatal falls overboard in Alaska between 1990 and 2005 were used. An in-depth descriptive analysis of these fatalities was performed to identify areas for intervention. RESULTS: There were 71 fatal falls overboard on commercial fishing vessels in Alaska during the 16 year time period. Falls overboard did not decline significantly during those years. The most common circumstances associated with falling overboard were working with fishing gear, being alone on deck, losing balance or slipping, heavy weather, gear entanglement, and alcohol. The level of involvement of those circumstances varied by region and gear type. SUMMARY: Many fatal falls overboard may be prevented by understanding the circumstances involved and targeting interventions at those specific risk factors. Interventions include creating more enclosed work spaces, managing lines, avoiding fishing alone, wearing personal flotation devices and man overboard alarms, and reducing alcohol use. Subsequent research should identify further interventions for each circumstance and evaluate the effectiveness of these interventions with the fishing industry. PMID- 17910032 TI - Treating young people with eating disorders: transition from child mental health to specialist adult eating disorder services. AB - BACKGROUND: The transition from Child and Adolescent Mental Health Services (CAMHS) to Adult Mental Health Services (AMHS) of young people with eating disorders may pose a number of difficulties, including an inconsistent referral process and age boundaries. METHODS: We compared young adults referred to a specialist Adult Eating Disorders Service (AEDS) who had previous involvement with CAMHS for the treatment of their eating disorder with those who did not. Information regarding the socio-demographic characteristics and eating disorders symptomatology of patients assessed by an AEDS over a 4-year period was collected. RESULTS: Patients who had previous involvement with CAMHS (particularly the ones treated as in-patients) presented with a lower self-esteem and more maturity fears (MF) than those without previous involvement. CONCLUSIONS: This study discusses the implication of these results in transitional arrangements between CAMHS and Adult services. It also highlights the need for heightened awareness of particular issues of self-esteem and maturation in these patients moving between services. PMID- 17910033 TI - Eating disorders and concurrent psychopathology: a reconceptualisation of clinical need through Rasch analysis. AB - Therapies for anorexia nervosa (AN), bulimia nervosa (BN), binge eating disorder (BED) and eating disorder not otherwise specified (EDNOS) take as their focus, perhaps of necessity, the eating disorder symptomatology. However, there is increasing evidence of comorbidity of psychopathological mechanisms (e.g. perfectionism, depression) with eating disorders which, left untreated, may diminish any therapeutic effects. Thus identifying the extent of comorbidity in an eating disorder population and assessing the relationship between psychopathological mechanisms and the eating disorder is important. Rasch analysis was applied to the findings from questionnaires (EDI-2; SCL90-R) completed by 105 female patients referred to an eating disorder unit. General psychopathology was found to be more indicative of 'caseness' than eating disorder psychopathology. In particular, interpersonal sensitivity, depression and mild interpersonal aspects of psychoticism emerged as important factors across eating disorders. The comorbidity of psychopathological mechanisms needs to be given consideration in the successful treatment of eating disorders. PMID- 17910034 TI - Regulation of C/EBPdelta-dependent transactivation by histone deacetylases in intestinal epithelial cells. AB - The C/EBPdelta transcription factor is involved in the positive regulation of the intestinal epithelial cell acute phase response. C/EBPdelta regulation by histone deacetylases (HDACs) during the course of inflammation remains to be determined. Our aim was to examine the effect of HDACs on C/EBPdelta-dependent regulation of haptoglobin, an acute phase protein induced in intestinal epithelial cells in response to pro-inflammatory cytokines. HDAC1, HDAC3, and HDAC4 were expressed in intestinal epithelial cells, as determined by Western blot. GST pull-down assays showed specific HDAC1 interactions with the transcriptional activation and the b ZIP C/EBPdelta domains, while the co-repressor mSin3A interacts with the C terminal domain. Immunoprecipitation assays confirmed the interaction between HDAC1 and the N-terminal C/EBPdelta amino acid 36-164 domain. HDAC1 overexpression decreased C/EBPdelta transcriptional activity of the haptoglobin promoter, as assessed by transient transfection and luciferase assays. Chromatin immunoprecipitation analysis showed a displacement of HDAC1 from the haptoglobin promoter in response to inflammatory stimuli and an increased acetylation of histone H3 and H4. HDAC1 silencing by shRNA expression increased both basal and IL-1beta-induced haptoglobin mRNA levels in epithelial intestinal cells. Our results suggest that interactions between C/EBPs and HDAC1 negatively regulate C/EBPdelta-dependent haptoglobin expression in intestinal epithelial cells. PMID- 17910035 TI - Non-conventional signal transduction by type 1 interferons: the NF-kappaB pathway. AB - Type I interferons (IFNs) regulate diverse cellular functions by modulating the expression of IFN-stimulated genes (ISGs) through the activation of the well established signal transduction pathway of the Janus Kinase (JAK) and signal transducers and activators of transcription (STAT) proteins. Although the JAK STAT signal transduction pathway is critical in mediating IFN's antiviral and antiproliferative activities, other signaling pathways are activated by IFNs and regulate cellular response to IFN. The NF-kappaB transcription factor regulates the expression of genes involved in cell survival and immune responses. We have identified a novel IFN mediated signal pathway that leads to NF-kappaB activation and demonstrate that a subset of ISGs that play key roles in cellular response to IFN is regulated by NF-kappaB. This review focuses on the IFN-induced NF-kappaB activation pathway and the role of NF-kappaB in ISG expression, antiviral activity and apoptosis, and the therapeutic application of IFN in cancer and infectious disease. PMID- 17910037 TI - Mother-offspring associations in northern muriquis, Brachyteles hypoxanthus. AB - Maternal care of offspring is ubiquitous among primates, but its duration varies across species due to factors such as dispersal patterns and social dynamics, which influence opportunities for and potential benefits of maternal investment in older offspring, respectively. We examined mother-offspring associations in wild northern muriquis (Brachyteles hypoxanthus), in which males are philopatric, females typically disperse before puberty, and social relationships among and between males and females are egalitarian. Associations were systematically recorded between ten mothers, each with two-six offspring in the study group, and all group members from August 2003-May 2004 at the RPPN-Feliciano Miguel Abdala in Minas Gerais, Brazil. Infants of both sexes received similarly high percentages of their mothers' association time. Mothers without infants also maintained strong associations with their youngest juvenile sons. Mothers did not spend consistently more time associating with either juvenile or adult sons than daughters. Our finding of non-preferential associations between muriqui mothers and their older male offspring suggests that extended maternal investment in offspring may be of minimal value in their egalitarian society compared with its value for species living in hierarchical societies. PMID- 17910036 TI - Deregulation of cofactor of BRCA1 expression in breast cancer cells. AB - Cofactor of BRCA1 (COBRA1) is an integral component of the human negative elongation factor (NELF), a four-subunit protein complex that inhibits transcription elongation. Previous in vivo work indicates that COBRA1 and the rest of the NELF complex repress estrogen-dependent transcription and the growth of breast cancer cells. In light of the COBRA1 function in breast cancer-related gene expression, we sought to examine regulation of COBRA1 expression in both established breast cancer cell lines and breast carcinoma tissues. We found that COBRA1 expression was inversely correlated with breast cancer progression, as tumor samples of patients who had distant metastasis and local recurrence expressed very low levels of COBRA1 mRNA when compared to those who were disease free for over 10 years (P = 0.0065 and 0.0081, respectively). Using both breast and prostate cancer cell lines, we also explored the possible mechanisms by which COBRA1 expression is regulated. Our results indicate that the protein abundance of COBRA1 and the other NELF subunits are mutually influenced in a tightly coordinated fashion. Small interfering RNA (siRNA) that targeted at one NELF subunit dampened the protein levels of all four subunits. Conversely, ectopic expression of COBRA1 in the knockdown cells partially rescues the co-depletion of the NELF subunits. In addition, our study suggests that a post-transcriptional, proteasome-independent mechanism is involved in the interdependent regulation of the NELF abundance. Furthermore, a lack of COBRA1 expression in breast carcinoma may serve as a useful indicator for poor prognosis. PMID- 17910039 TI - Glucocorticoid-induced leucine zipper (GILZ) mediates glucocorticoid action and inhibits inflammatory cytokine-induced COX-2 expression. AB - Cyclooxygenase-2 (COX-2) plays an important role in rheumatoid arthritis and therefore, has been a major target for anti-arthritis therapies. The expression of COX-2 is induced by inflammatory cytokines such as TNF-alpha and IL-1beta, and inhibited by glucocorticoids. However, the molecular mechanisms underlying the anti-inflammatory and immune suppressive actions of glucocorticoids are not well defined. Here we report that glucocorticoid-induced leucine zipper (GILZ) mimics glucocorticoid action and inhibits inflammatory cytokine-induced COX-2 expression in bone marrow mesenchymal stem cells, the cells that have been recently implicated in the pathogenesis and progression of rheumatoid arthritis. Using a retrovirus-mediated gene expression approach we demonstrate that overexpression of GILZ inhibits TNF-alpha and IL-1beta-induced COX-2 mRNA and protein expression, and knockdown of GILZ by shRNA reduces glucocorticoid inhibition of cytokine-induced COX-2 expression. Consistent to these results, overexpression of GILZ also inhibits NF-kappaB-mediated COX-2 promoter activity. Finally, we show that GILZ inhibits COX-2 expression by blocking NF-kappaB nuclear translocation. Our results suggest that GILZ is a key glucocorticoid effect mediator and that GILZ may have therapeutic value for novel anti-inflammation therapies. PMID- 17910040 TI - Prolonged half-life of argatroban in patients with renal dysfunction and antiphospholipid antibody syndrome being treated for heparin-induced thrombocytopenia. AB - Argatroban is a direct thrombin inhibitor approved for the treatment of heparin induced thrombocytopenia (HIT) type II. Argatroban is predominantly metabolized in the liver. It is widely believed that no dosage adjustment is required in patients with renal insufficiency, making it a preferred agent in patients on renal replacement therapy (Reddy and Grossman, Ann Pharm 2005;39:1601-1605). The elimination half-life of argatroban is approximately 50 min. Lupus anticoagulants can cause baseline elevation of the PTT and hence it is difficult to monitor the effects of anticoagulants such as heparin, lepirudin, or argatroban in patients with antiphospholipid antibody syndrome. Heparin levels may be used as an alternative for heparin monitoring but plasma levels of argatroban are not commercially available. A chromogenic antifactor IIa assay could be useful for monitoring argatroban in the presence of a lupus anticoagulant, but it is not widely available at present. We report a patient with end-stage renal disease, maintained on peritoneal dialysis with HIT, who demonstrated a markedly prolonged half-life when treated with argatroban despite the discontinuation of therapy. This case also demonstrates the lack of guidelines for the monitoring of argatroban therapy in the presence of an underlying lupus anticoagulant. PMID- 17910038 TI - Generation of novel, secreted epidermal growth factor receptor (EGFR/ErbB1) isoforms via metalloprotease-dependent ectodomain shedding and exosome secretion. AB - Exosomes are small membrane vesicles derived from intracellular multivescicular bodies (MVBs) that can undergo constitutive and regulated secretion from cells. Exosomes can also secrete soluble proteins through metalloprotease-dependent ectodomain shedding. In this study, we sought to determine whether ErbB1 receptors are present within exosomes isolated from the human keratinocyte cell line, HaCaT, and whether exosome-associated ErbB1 receptors can undergo further proteolytic processing. We show that full-length transmembrane ErbB1 is secreted in HaCaT exosomes. EGF treatment and calcium flux stimulated the release of phosphorylated ErbB1 in exosomes but only ligand-stimulated release was blocked by the ErbB1 kinase inhibitor, AG1478, indicating that ligand-dependent ErbB1 receptor activation can initiate ErbB1 secretion into exosomes. In addition, other immunoreactive but truncated ErbB1 isoforms were detected in exosomes suggestive of additional proteolytic processing. We demonstrate that cellular and exosomal ErbB1 receptors can undergo ectodomain shedding to generate soluble N terminal ectodomains and membrane-associated C-terminal remnant fragments (CTFs). ErbB1 shedding was activated by calcium flux and the metalloprotease activator APMA (4-aminophenylmercuric acetate) and was blocked by a metalloprotease inhibitor (GM6001). Soluble ErbB1 ectodomains shed into conditioned medium retained the ability to bind exogenous ligand. Our results provide new insights into the proteolysis, trafficking and fate of ErbB1 receptors and suggest that the novel ErbB1 isoforms may have functions distinct from the plasma membrane receptor. PMID- 17910041 TI - Utility of consecutive repeat HIT ELISA testing for heparin-induced thrombocytopenia. AB - Heparin-induced thrombocytopenia (HIT) is a serious complication of heparin therapy. Limited data are available regarding repeat HIT antibody testing after an initial negative test. We conducted a retrospective study to determine the utility of repeat testing. Heparin antibodies were detected using the GTI-PF4 enzyme-linked immunoabsorbent assay, ELISA (GTI Diagnostics, Waukesha, WI). Patients (n = 137) were assigned to one of three groups based upon the initial negative test optical density (OD) range of low = 0-0.132, medium = 0.133-0.267, and high = 0.268-0.399. A pretest clinical score was retrospectively determined using the "4T's" (Thrombocytopenia, Timing of platelet fall, Thrombosis, and the absence of oTher causes of thrombocytopenia). A subsequent positive ELISA was found in 16% (22/137) of patients who underwent repeat testing. Most of these patients had a low pretest clinical score (62%). Four patients had an interval change in the pretest score between the initial negative and subsequent positive tests. Only these four patients developed HIT with thrombosis (HITT). Eighty percent of patients with a high initial negative test OD value had a positive ELISA on repeat testing; however, the initial negative test OD value could not predict whether a patient developed HITT. In contrast, an increase in the pretest clinical probability between initial and repeat testing better predicted HITT. Consecutive repeat ELISA testing for heparin antibodies may be warranted in patients with an increase in their pretest clinical score after an initial negative test as an adjunct to confirm the diagnosis of HIT. PMID- 17910042 TI - New polymorphism in the NCF-2 gene leading to alternative splicing without altering gene expression or the respiratory burst activity. PMID- 17910043 TI - Identification of genes with abnormal expression changes in acute myeloid leukemia. AB - Acute myeloid leukemia (AML) is one of the most common and deadly forms of hematopoietic malignancies. We hypothesized that microarray studies could identify previously unrecognized expression changes that occur only in AML blasts. We were particularly interested in those genes with increased expression in AML, believing that these genes may be potential therapeutic targets. To test this hypothesis, we compared gene expression profiles between normal hematopoietic cells from 38 healthy donors and leukemic blasts from 26 AML patients. Normal hematopoietic samples included CD34+ selected cells (N = 18), unselected bone marrows (N = 10), and unselected peripheral bloods (N = 10). Twenty genes displayed AML-specific expression changes that were not found in the normal hematopoietic cells. Subsequent analyses using microarray data from 285 additional AML patients confirmed expression changes for 13 of the 20 genes. Seven genes (BIK, CCNA1, FUT4, IL3RA, HOMER3, JAG1, WT1) displayed increased expression in AML, while 6 genes (ALDHA1A, PELO, PLXNC1, PRUNE, SERPINB9, TRIB2) displayed decreased expression. Quantitative RT/PCR studies for the 7 over expressed genes were performed in an independent set of 9 normal and 21 pediatric AML samples. All 7 over-expressed genes displayed an increased expression in the AML samples compared to normals. Three of the 7 over-expressed genes (WT1, CCNA1, and IL3RA) have already been linked to leukemogenesis and/or AML prognosis, while little is known about the role of the other 4 over-expressed genes in AML. Future studies will determine their potential role in leukemogenesis and their clinical significance. PMID- 17910044 TI - Sickle cell disease and pulmonary hypertension in Africa: a global perspective and review of epidemiology, pathophysiology, and management. AB - Secondary pulmonary hypertension (PAH) has been shown to have a prevalence of 30% in patients with sickle cell disease (SCD) with mortality rates of 40% at 40 months after diagnosis in the United States. The burden of SCD is highest in sub Saharan Africa, especially in Nigeria (West Africa), where approximately 6 million people are afflicted. The true global incidence, prevalence, and burden of SCD and its associated end organ complications however remain unknown. Chronic hemolysis represents a prominent mechanistic pathway in the pathogenesis of SCD associated pulmonary hypertension via a nitric oxide (NO) scavenging and abrogation of NO salutatory effects on vascular function, including smooth muscle relaxation, downregulation of endothelial adhesion molecules and inhibition of platelet activation. Many known infectious risk factors for PAH are also hyperendemic in Africa, including Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome (HIV/AIDS), chronic hepatitis B and C, and possibly malaria. Interactions between these infectious complications and SCD-related hemolysis could yield an even higher prevalence of pulmonary hypertension and compound the existing global health systems challenges in managing SCD. Indeed, our preliminary analysis of African immigrants currently in the United States suggests that pulmonary hypertension represents a significant complication of SCD in the African subcontinent. There is clearly a need to include Africa and other parts of the world with high SCD prevalence in future comprehensive studies on the epidemiology and treatment of end organ complications of an aging SCD population world-wide. PMID- 17910045 TI - Mutations of FLT3, NRAS, KRAS, and PTPN11 are frequent and possibly mutually exclusive in high hyperdiploid childhood acute lymphoblastic leukemia. AB - Although it has been suggested that mutations of the FLT3, NRAS, KRAS, and PTPN11 genes are particularly frequent in high hyperdiploid (>50 chromosomes) pediatric acute lymphoblastic leukemias (ALLs), this has as yet not been confirmed in a large patient cohort. Furthermore, it is unknown whether mutations of these genes coexist in hyperdiploid cases. We performed mutation analyses of FLT3, NRAS, KRAS, and PTPN11 in a consecutive series of 78 high hyperdiploid ALLs. Twenty-six (33%) of the cases harbored a mutation, comprising six activating point mutations and one internal tandem duplication of FLT3 (7/78 cases; 9.0%), eight codon 12, 13, or 61 NRAS mutations (8/78 cases; 10%), five codon 12 or 13 KRAS mutations (5/78 cases, 6.4%), and seven exon 3 or 13 PTPN11 mutations (7/78 cases; 9.0%). No association was seen between the presence of a mutation in FLT3, NRAS, KRAS, or PTPN11 and gender, age, white blood cell count, or relapse, suggesting that they do not confer a negative prognostic impact. Only one case harbored mutations in two different genes, suggesting that mutations of these four genes are generally mutually exclusive. In total, one third of the cases harbored a FLT3, NRAS, KRAS, or PTPN11 mutation, identifying the RTK-RAS signaling pathway as a potential target for novel therapies of high hyperdiploid pediatric ALLs. PMID- 17910046 TI - Design, synthesis, and metal binding of novel Pseudo- oligopeptides containing two phosphinic acid groups. AB - Phosphinic compounds have potential as amide-bond mimetics in the development of novel peptidomimetics, enzyme inhibitors, and metal-binding ligands. Novel pseudo oligopeptides with two phosphinic acid groups embedded in the peptide backbone serving as amide-bond surrogates, Psi[P(O,OH)--CH(2)], were targeted. A series of linear and cyclic pseudo-oligopeptides with two phosphinic acid groups arrayed at different positions in the peptide sequence were designed, including Ac--Phe- {(R,S)--AlaPsi[P(O,OH)--CH(2)]Gly}(2)--NH(2) (P2), Ac--NH--(R,S)--AlaPsi[P(O,OH)- CH(2)]Gly--Phe--(R,S)--AlaPsi[P(O,OH)--CH(2)]Gly--NH(2) (P3), Ac--NH--(R,S)- AlaPsi[P(O,OH)--CH(2)]Gly--Phe--Phe--(R,S) --AlaPsi[P(O,OH)--CH(2)]Gly--NH(2) (P4), cyclo{NH--(R,S)--AlaPsi[P(O,OH)--CH(2)]Gly--Phe}(2) (P5), and cyclo[NH- (R,S)--AlaPsi[P(O,OH)--CH(2)]Gly--Phe--Phe](2) (P6). They were synthesized via conventional Fmoc chemistry on solid support utilizing Fmoc-protected phosphinic acid-containing pseudo-dipeptide fragment, i.e. Fmoc--(R,S)--AlaPsi[P(O,OCH(3))- CH(2)]Gly--OH. The pseudo-peptides containing two phosphinic acid groups exhibited the highest binding affinity and selectivity for Fe(III) among the 10 metal ions screened by ESI-MS analysis--Cu(II), Zn(II), Co(II), Ni(II), Mn(II), Fe(II), Fe(III), Al(III), Ga(III), and Gd(III). P4 and P6 with 11-atom linkages between the two phosphinic acids preferred intramolecular metal binding to form 1:1 ligand/metal complexes. As revealed by competition experiments, P4 showed the highest relative binding affinity among the six compounds tested. Noteworthy, P4 also showed higher relative binding affinity than similar dihydroxamate containing pseudo-peptides reported previously. The novel structural prototype and facile synthesis along with selective and potent Fe(III) binding strongly suggest that pseudo-peptides containing the two or more phosphinic groups as amide-bond surrogates deserve further exploration in medicinal chemistry. PMID- 17910047 TI - Mechanical testing of fixation techniques for scaffold-based tissue-engineered grafts. PMID- 17910048 TI - Removal of dentin matrix proteoglycans by trypsin digestion and its effect on dentin bonding. AB - The aim of this study was to investigate the effect of trypsin digestion on removal of chondroitin sulfate proteoglycans (CS-PGs) in demineralized dentin, and subsequent dentin bonding. Bovine dentin fragments were demineralized, treated with or without trypsin, stained with cupromeronic blue, and observed under transmission electron microscopy. Demineralized sections with or without trypsin digestion were also subjected to immunohistochemical analysis with anti chondroitin-4-sulfate (C4S) monoclonal antibody, 2-B-6. The presence of galactosamine and glucosamine in the trypsin digest was confirmed by amino acid analysis. Bond strength testing was performed on trypsin treated and control specimens where samples were either kept moist or dried and re-wet, then bonded. Bond strength significantly decreased after trypsin treatment (p < 0.05). TEM, immunohistochemical, and amino acid analyses demonstrated that trypsin digestion efficiently removed C4S-PGs from demineralized dentin matrix. This study indicates that the detrimental effects observed on dentin bonding by trypsinization may be due in part to the removal/cleavage of the C4S-PGs, and further underscore the importance of C4S-PGs on dentin bonding. PMID- 17910050 TI - Sub-micromolar increase in [Ca(2+)](i) triggers delayed exocytosis of ATP in cultured astrocytes. AB - Astrocytes release a variety of transmitter molecules, which mediate communication between glial cells in the brain and modulate synaptic transmission. ATP is a major glia-derived transmitter, but the mechanisms and kinetics of ATP release from astrocytes remain largely unknown. Here, we combined epifluorescence and total internal reflection fluorescence microscopy to monitor individual quinacrine-loaded ATP-containing vesicles undergoing exocytosis in cultured astrocytes. In resting cells, vesicles exhibited three-dimensional motility, spontaneous docking and release at low rate. Extracellular ATP application induced a Ca(2+)-dependent increase in the rate of exocytosis, which persisted for several minutes. Using UV flash photolysis of caged Ca(2+), the threshold [Ca(2+)](i) for ATP exocytosis was found to be approximately 350 nM. Subthreshold [Ca(2+)](i) transients predominantly induced vesicle docking at plasma membrane without subsequent release. ATP exocytosis triggered either by purinergic stimulation or by Ca(2+) uncaging occurred after a substantial delay ranging from tens to hundreds of seconds, with only approximately 4% of release occurring during the first 30 s. The time course of the cargo release from vesicles had two peaks centered on 3 Mb. The 9qSTDS is caused by haplo-insufficiency of EHMT1, a gene whose protein product (Eu-HMTase1) is a histone H3 Lys 9 (H3-K9) methyltransferase. This was established by identification of three patients with features of the syndrome and either mutations or a balanced translocation in EHMT1. H3-K9 histone methylation is restricted to the euchromatin of mammals and functions to silence individual genes. Deletion size does not correlate with the severity of the 9qSTDS since patients with mutations in EHMT1 are as severely affected as those with submicroscopic deletions. Patients clinically suspected of having the 9qSTDS but with normal subtelomere deletion testing by FISH or MLPA should be considered for detailed 9q MLPA analysis and/or sequencing of EHMT1. EHMT1 is another example in the growing list of genes responsible for brain development that appear to play a role in chromatin remodeling. Published 2007 Wiley-Liss, Inc. PMID- 17910073 TI - Cryptic telomere imbalance: a 15-year update. AB - It has been 15 years since we proposed that assays of telomere integrity might reveal cryptic translocations and deletions as a significant cause of mental retardation (MR) in patients with normal G-banded karyotypes. Development of unique genomic probes adjacent to the subtelomeric repeats of each chromosome arm allowed multiplex FISH analyses that confirmed such cryptic telomeric imbalances in 3-6% of all unexplained MR. Although such "telomere FISH" analysis quickly became standard of care, limitations of this technology platform included a lack of information on the size and gene content of the deleted/duplicated segments and the failure to detect interstitial deletions not involving the most distal unique clone. The development of "molecular ruler" clone sets for every human telomere provided the foundation for accurate determination of size and gene content of each imbalance, as well as the detection of interstitial deletions within these regions. Array comparative genomic hybridization (aCGH) has emerged as a powerful technology to assess single copy changes (monosomy or trisomy) at targeted loci such as telomeres or across the whole genome. This technology now replaces multiplex FISH for the assessment of telomere integrity in unexplained MR and has the advantage of efficiently determining the size and gene content of the imbalance, as well as detecting interstitial deletions near telomeres or anywhere else in the genome covered by the array design. The application of aCGH in several studies of unexplained MR has confirmed that telomere imbalances are overrepresented compared to "average" chromosomal regions, although this is likely due to random chromosome breakage rather than specific molecular mechanisms associated with the genomic architecture of human telomeres. Telomere imbalances are significantly larger than initially envisioned ( approximately 40% are >5 Mb in size), and indicate the analytic sensitivity of the G-banded karyotype is much lower than previously thought. Finally, experience with smaller benign variants compared to larger pathogenic imbalances at telomeres serves as a model for approaching whole-genome aCGH in a clinical setting. PMID- 17910074 TI - Novel microdeletion syndromes. PMID- 17910075 TI - Chromosome 5q subtelomeric deletion syndrome. AB - The pure 3.5 Mb subtelomeric deletion syndrome is very rare but causes a recognizable phenotype characterized by prenatal lymphedema with increased nuchal translucency, pronounced muscular hypotonia in infancy, borderline intelligence, postnatal short stature with delayed bone age due to growth hormone deficiency, and multiple minor anomalies including mildly bell-shaped chest, minor congenital heart defects, and a distinct facial gestalt. Terminal deletions including the adjacent approximately 2 Mb NSD1-locus show a compound phenotype with overlap to Sotos syndrome. Larger terminal deletions including also chromosomal bands 5q35.1 and 5q35.2 cause a more severe phenotype with normal body length, significant congenital heart defect, microcephaly, profound developmental retardation or early death due to respiratory failure. Heart defects in the latter are explained by haploinsufficiency of the NKX2.5 gene at 5q35.1. The deletion breakpoint of the 3.5 Mb subtelomeric microdeletion maps to a low copy repeat which is identical to the distal copy of two highly similar regions flanking the recurrent interstitial NSD1 microdeletion. As meiotic mispairing between these low copy repeats seem to be much more likely than a terminal aberration, these neighborhood may prevent occurrence of the subtelomeric deletion syndrome, which could explain the rareness of the latter. PMID- 17910076 TI - The identification of microdeletion syndromes and other chromosome abnormalities: cytogenetic methods of the past, new technologies for the future. AB - Chromosome analysis is an important diagnostic tool in the identification of causes of mental retardation, developmental delay, and other developmental disabilities. Cytogenetic approaches have revealed the chromosomal basis of a large number of genetic syndromes. The recent use of microarray-based comparative genomic hybridization (array CGH) has accelerated the identification of novel cytogenetic abnormalities. We present the results of array CGH in 8,789 clinical cases submitted for a variety of developmental problems. Of these cases, 6.9% showed clinically relevant abnormalities, 1.2% showed benign copy-number variants (polymorphisms), 2.5% showed recurrent alterations of unclear clinical significance-many of which are likely to be polymorphisms-and 1.4% showed novel alterations of unclear relevance. Although cytogenetic methods, including array CGH, have great potential for identifying novel chromosomal syndromes, this high resolution analysis may also result in diagnostic challenges imposed on laboratories and clinicians regarding findings of unclear clinical significance. (c) 2007 Wiley-Liss, Inc. PMID- 17910077 TI - Chromosome 2q37 deletion: clinical and molecular aspects. AB - Terminal deletions of chromosome 2 with breakpoints at or within band 2q37, ranging from visible abnormalities to cryptic, subtelomeric deletions, have been recognized with increasing frequency among children with mild-moderate mental retardation, characteristic facial appearance, and behavioral manifestations which often place them on the autism spectrum. The stereotypic facial characteristics include prominent forehead, thin, highly arched eyebrows, depressed nasal bridge, full cheeks, deficient nasal alae and prominent columella, thin upper lip, and various minor anomalies of the pinnae. Abnormal nipples, including inverted nipples, have been reported in a number of cases. CNS, ocular, cardiac, gastrointestinal, renal, and other GU anomalies have been noted in nearly one-third of patients. Of note, coarctation or hypoplasia of the aorta has been described in several affected children. Wilms tumor, renal dysplasia, and tracheomalacia have been reported only with the most proximal breakpoint at band 2q37.1 while a range of GI anomalies, pyloric stenosis, and diaphragmatic defects have been reported with breakpoints throughout the region. A subset of patients with the most distal deletion present phenotypic features which mimic Albright hereditary osteodystrophy (AHO). In addition to the AHO-like phenotype, later onset findings include seizures and cystic kidneys. Timely diagnosis of this recognizable syndrome provides a basis for genetic counseling, appropriate surveillance, and intervention, and avoids unnecessary and expensive diagnostic testing. PMID- 17910078 TI - Estrogen effect in multiple sclerosis more nuanced than described. PMID- 17910079 TI - Education and children with Down syndrome: neuroscience, development, and intervention. AB - Of the recent advances in education-related research in Down syndrome, the characterization of the Down syndrome behavioral phenotype has become a potentially critical tool for shaping education and intervention in this population. This article briefly reviews the literature on brain-behavior connections in Down syndrome and identifies aspects of the Down syndrome behavioral phenotype that are potentially relevant to educators. Potential challenges to etiologically informed educational planning are discussed. PMID- 17910080 TI - Psychiatric and behavioral disorders in persons with Down syndrome. AB - Similar to the state of the broader intellectual disabilities field, many gaps exist in the research and treatment of mental health concerns in people with Down syndrome. This review summarizes key findings on the type and prevalence of behavior and emotional problems in children, adolescents, and adults with Down syndrome. Such findings include relatively low rates of severe problems in children, and well-documented risks of depression and Alzheimer's disease in older adults. The review also considers emerging data on autism, and the paucity of studies on adolescents. Three next steps for research are highlighted, including a need to: (1) connect research on psychiatric status and diagnoses across developmental periods, including adolescence, and to examine such associated processes as sociability, anxiety and attention; (2) unravel complicated biopsycho-social risk and protective factors that serve to increase or diminish psychopathology; and (3) identify evidence-based treatments that both reduce distressful symptoms and enhance well-being in individuals with Down syndrome. PMID- 17910081 TI - Families of persons with Down syndrome: new perspectives, findings, and research and service needs. AB - To understand the families of offspring with Down syndrome, this article begins by describing the change in orientation--from "negative" to "stress-and-coping" perspectives--in studies of families of offspring with disabilities. In reviewing the existing studies, mothers, fathers, and siblings cope slightly better than family members of persons with other disabilities, a phenomenon called the "Down syndrome advantage." Beyond this more general finding, however, much remains unknown. Most studies examine only parental or sibling levels of stress or coping, leaving unknown the marital, occupational, health, educational, and other "real-world" outcomes for these family members. Increased research attention is needed to understand the life-span needs of families of persons with Down syndrome and the impact of cultural and sociocultural diversity on family outcomes. It will also be important to relate family outcomes to differences in the offspring's behaviors, development, relationships, medical conditions, psychopathology, and the presence (and effectiveness) of needed support services. Although these families have received some research attention over the past several decades, we now need to make family research in Down syndrome more concrete, more life-span, and more tied to characteristics of the individual with the syndrome and the family's surrounding support system. PMID- 17910083 TI - A year of unprecedented progress in Down syndrome basic research. AB - The years 2006 and 2007 saw the publication of three new and different approaches to prevention or amelioration of Down syndrome effects on the brain and cognition. We describe the animal model systems that were critical to this progress, review these independent breakthrough studies, and discuss the implications for therapeutic approaches suggested by each. PMID- 17910084 TI - Down syndrome: cognitive phenotype. AB - Down syndrome is the most prevalent cause of intellectual impairment associated with a genetic anomaly, in this case, trisomy of chromosome 21. It affects both physical and cognitive development and produces a characteristic phenotype, although affected individuals vary considerably with respect to severity of specific impairments. Studies focusing on the cognitive characteristics of Down syndrome were reviewed, and while performance in most areas could be predicted based upon overall intellectual disability, relative weaknesses were consistently found to be associated with expressive language, syntactic/morphosyntactic processing, and verbal working memory. This profile of uneven deficits could result from a failure to develop typically automatic processing for speech perception and production, and this possibility is discussed along with its implications for intervention. PMID- 17910085 TI - Alzheimer's disease in Down syndrome: neurobiology and risk. AB - Down syndrome (DS) is characterized by increased mortality rates, both during early and later stages of life, and age-specific mortality risk remains higher in adults with DS compared with the overall population of people with mental retardation and with typically developing populations. Causes of increased mortality rates early in life are primarily due to the increased incidence of congenital heart disease and leukemia, while causes of higher mortality rates later in life may be due to a number of factors, two of which are an increased risk for Alzheimer's disease (AD) and an apparent tendency toward premature aging. In this article, we describe the increase in lifespan for people with DS that has occurred over the past 100 years, as well as advances in the understanding of the occurrence of AD in adults with DS. Aspects of the neurobiology of AD, including the role of amyloid, oxidative stress, Cu/ZN dismutase (SOD-1), as well as advances in neuroimaging are presented. The function of risk factors in the observed heterogeneity in the expression of AD dementia in adults with DS, as well as the need for sensitive and specific biomarkers of the clinical and pathological progressing of AD in adults with DS is considered. PMID- 17910086 TI - Genetic mechanisms involved in the phenotype of Down syndrome. AB - Down syndrome (DS) is the most common genetic cause of significant intellectual disability in the human population, occurring in roughly 1 in 700 live births. The ultimate cause of DS is trisomy of all or part of the set of genes located on chromosome 21. How this trisomy leads to the phenotype of DS is unclear. The completion of the DNA sequencing and annotation of the long arm of chromosome 21 was a critical step towards understanding the genetics of the phenotype. However, annotation of the chromosome continues and the functions of many genes on chromosome 21 remain uncertain. Recent findings about the structure of the human genome and of chromosome 21, in particular, and studies on mechanisms of gene regulation indicate that various genetic mechanisms may be contributors to the phenotype of DS and to the variability of the phenotype. These include variability of gene expression, the activity of transcription factors both encoded on chromosome 21 and encoded elsewhere in the genome, copy number polymorphisms, the function of conserved nongenic regions, microRNA activities, RNA editing, and perhaps DNA methylation. In this manuscript, we describe current knowledge about these genetic complexities and their likely importance in the context of DS. We identify gaps in current knowledge and suggest priorities to fill these gaps. PMID- 17910088 TI - Toward a research agenda for Down syndrome. PMID- 17910087 TI - Language development in Down syndrome: from the prelinguistic period to the acquisition of literacy. AB - Down syndrome (DS) is associated with abnormalities in multiple organ systems and a characteristic phenotype that includes numerous behavioral features. Language, however, is among the most impaired domains of functioning in DS and, perhaps, also the greatest barrier to independent meaningful inclusion in the community. In this article, we review what is known about the extent, nature, and correlates of the language and related problems of individuals with Down syndrome. In doing so, we focus largely on the syndrome-specific features of the language phenotype, although we also consider within-syndrome variation. The review focuses on the prelinguistic foundations of language and the major components of language (i.e., vocabulary, syntax, and pragmatics). We also consider two topics in the treatment and education of individuals with DS: prelinguistic communication intervention and the acquisition of literacy skills. PMID- 17910090 TI - Epidemiology of Down syndrome. AB - Down syndrome (DS) is the most commonly identified genetic form of mental retardation and the leading cause of specific birth defects and medical conditions. Traditional epidemiological studies to determine the prevalence, cause, and clinical significance of the syndrome have been conducted over the last 100 years. DS has been estimated to occur in approximately 1 in 732 infants in the United States, although there is some evidence that variability in prevalence of estimates exist among racial/ethnic groups. Progress has been made in characterizing the specific types of chromosome errors that lead to DS and in identifying associated factors that increase the risk of chromosome 21 malsegregation, i.e., advanced maternal age and recombination. Studies to examine the variability of the presence of specific DS-associated birth defects and medical conditions provide evidence for genetic and environmental modifiers. Here, we provide a brief survey of studies that address the current state of the field and suggest gaps in research that can soon be filled with new multidisciplinary approaches and technological advances. PMID- 17910089 TI - Using mouse models to explore genotype-phenotype relationship in Down syndrome. AB - Down Syndrome (DS) caused by trisomy 21 is characterized by a variety of phenotypes and involves multiple organs. Sequencing of human chromosome 21 (HSA21) and subsequently of its orthologues on mouse chromosome 16 have created an unprecedented opportunity to explore the complex relationship between various DS phenotypes and the extra copy of approximately 300 genes on HSA21. Advances in genetics together with the ability to generate genetically well-defined mouse models have been instrumental in understanding the relationships between genotype and phenotype in DS. Indeed, elucidation of these relationships will play an important role in understanding the pathophysiological basis of this disorder and helping to develop therapeutic interventions. A successful example of using such a strategy is our recent studies exploring the relationship between failed nerve growth factor (NGF) transport and amyloid precursor protein (App) overexpression. We found that increased dosage of the gene for App is linked to failed NGF signaling and cholinergic neurodegeneration in a mouse model of DS. Herein, we discuss several mouse models of DS and explore the emergence of exciting new insights into genotype-phenotype relationships, particularly those related to nervous system abnormalities. An important conclusion is that uncovering these relationships is enhanced by working from carefully defined phenotypes to the genes responsible. PMID- 17910092 TI - Electrospun chitosan-coated fibers of poly(L-lactide) and poly(L lactide)/poly(ethylene glycol): preparation and characterization. AB - Fibrous poly(L-lactide) (PLLA) and bicomponent PLLA/poly(ethylene glycol) mats were prepared by electrospinning and then were coated with chitosan. The presence of chitosan coating was proved by scanning electron microscopy and by fluorescence microscopy. On contact with blood, the chitosan coating led to changes in erythrocyte shape and in their aggregation. The haemostatic activity of the mats increased with increasing chitosan content. Microbiological studies against Staphylococcus aureus revealed that the chitosan coating imparts antibacterial activity to the hybrid mats. The combined haemostatic and antibacterial activities render these novel materials suitable for wound-healing applications. PMID- 17910091 TI - Preparation and biodegradation of sugar-containing poly(vinyl acetate) emulsions. AB - To accelerate the biodegradability of poly(vinyl acetate)-based emulsions, emulsion copolymerizations of vinyl sugars, including triacetylated N-acetyl-D glucosamine (GlcNAc)-substituted 2-hydroxyethyl methacrylate (GlcNAc(Ac)3 substituted HEMA), glucose-substituted HEMA (GEMA) and 6-O-vinyladipoyl-D-glucose (6-O-VAG) with vinyl acetate (VAc), were carried out using poly(vinyl alcohol) as an emulsifying agent in the presence of poly[(butylene succinate)-co-(butylene adipate)] [poly(BS-co-BA)]. Copolymerization with GEMA produced a stable emulsion and that with 6-O-VAG also produced a homogeneous emulsion. Their biodegradation tests indicated that PVAc main chain scission was accelerated by copolymerization with vinyl sugars. PMID- 17910093 TI - Development of nanostructure in resistant starch type III during thermal treatments and cycling. AB - The effect of recrystallization temperature on the lamellar structure of RSIII samples was studied using XRD and SAXS. The polymorph type could be manipulated in a controlled manner, independently of the plant source. In RSIII from corn starch and from high-amylose corn starch, retrogradation at a low temperature led to the formation of polymorph B with lamellas arranged in long-range periodicity, whereas retrogradation at a high temperature led to the formation of polymorphs A and V with no defined periodicity. The retrogradation temperature of wheat starch did not have a major effect on its nanostructure. For both polymorphs, the enzymatic degradation decreased as the degree of order within the crystal increased. PMID- 17910094 TI - Giving greater financial independence to hospitals--does it make a difference? The case of English NHS Trusts. AB - In 2003 a new type of provider organisation, the Foundation Trust (FT), was introduced in England, and the best performing NHS hospitals were able to apply for 'Foundation status'. FTs enjoy greater financial flexibility and are subject to less central monitoring and control. The phased introduction of FTs represents an opportunity to examine whether the new financial structures facing FTs have produced any differences in financial performance compared with non-FTs. We use difference in difference methods to examine whether Foundation status had a significant effect on financial management. We find that Foundation status has had a limited impact in terms of acting as an instrument to signal strong financial management of FTs. This result may reflect the relatively early stage of the FT process or may be due to the fact that all types of Trusts are experiencing a challenging financial environment, including the introduction of a prospective payment system. However, we explore the nature of the trends emerging over time and discuss the implications of our findings for policy. PMID- 17910095 TI - Methanol production from CO(2) by resting cells of the methanotrophic bacterium Methylosinus trichosporium IMV 3011. AB - Methanol production from carbon dioxide was successfully achieved using resting cells of Methylosinus trichosporium IMV 3011 as biocatalysts. Carbon dioxide was reduced to methanol and extracellular methanol accumulation has been found in the carbon dioxide incubations. However, resting cells of methanotrophs have a finite or intrinsic methanol production capacity due to a limiting supply of intracellular reducing equivalent. It has been found that the catabolism of stored Poly-beta -Hydroxybutyrate (PHB) can provide intracellular reducing equivalents to improve the intrinsic methanol production capacity. The initial nitrogen and copper concentration in the culture medium were studied for the accumulation of PHB by M. trichosporium IMV 3011, to expand its potential uses in methanol production from carbon dioxide reduction. It appeared that the total methanol production capacity was increased with increasing PHB content in cells. Resting cells containing 38.6% PHB exhibited the highest total methanol production capacity. But higher PHB accumulation adversely affected the total methanol production capacity. The effects of methanol production process on the survival and recovery of M. trichosporium IMV 3011 were examined. The results showed that the methanol production from carbon dioxide reduction was not detrimental to the viability of methanotrophs. PMID- 17910096 TI - Establishment of Azotobacter on plant roots: chemotactic response, development and analysis of root exudates of cotton (Gossypium hirsutum L.) and wheat (Triticum aestivum L.). AB - Biofertilizers contribute in N(2) fixation, P solubilization, phytohormone production and thus enhance plant growth. Beneficial plant-microbe interactions and the stability and effectiveness of biofertilizer depend upon the establishment of bacterial strains in the rhizosphere of the plant. This interaction depends upon many factors, one of them being plant exudates. Root exudates are composed of small organic molecules like carbonic acids, amino acids or sugars etc., which are released into the soil and bacteria can be attracted towards these exudates due to chemotaxis. The chemotactic behaviour of Azotobacter strains was studied using cotton (Desi HD 123 and American H 1098) and wheat (WH 711) seedlings and the root exudates of these two plants were chemically characterized. Analysis of the root exudates revealed the presence of sugars and simple polysaccharides (glucose), amino acids (glutamate, lysine) and organic acids (citric acid, succinic acid, maleic acid, malonic acid). Differences between cotton cultivars in root exudates were observed which influenced chemotactic response in Azotobacter. These results indicate colonization with rhizobacteria which implies that optimal symbionts, on the sides of both plant cultivar and bioinoculant bacteria can lead to better plant growth under cultivation conditions. PMID- 17910097 TI - Regulation of pyrimidine formation in Pseudomonas oryzihabitans. AB - The regulation of pyrimidine formation in the opportunistic human pathogen Pseudomonas oryzihabitans was investigated at the level of enzyme synthesis and at the level of activity for the pyrimidine biosynthetic pathway enzyme aspartate transcarbamoylase. Although pyrimidine supplementation of succinate-grown P. oryzihabitans cells produced little effect on the de novo pyrimidine biosynthetic pathway enzyme activities, pyrimidine limitation experiments undertaken using an orotidine 5'-monophosphate decarboxylase mutant strain isolated from P. oryzihabitans ATCC 43272 indicated that repression of enzyme synthesis by pyrimidines was occurring. Following pyrimidine limitation of the succinate-grown decarboxylase mutant strain cells, aspartate transcarbamoylase and dihydroorotase activities were found to increase by about 3-fold while dihydroorotate dehydrogenase and orotate phosphoribosyltransferase activities were also observed to increase relative to their activities in the mutant strain cells grown on excess uracil. At the level of enzyme activity, aspartate transcarbamoylase in P. oryzihabitans was strongly inhibited by pyrophosphate, ADP, ATP and GTP in the presence of saturating substrate concentrations. PMID- 17910100 TI - Mutations and environmental factors affecting regulation of riboflavin synthesis and iron assimilation also cause oxidative stress in the yeast Pichia guilliermondii. AB - Iron deficiency causes oversynthesis of riboflavin in several yeast species, known as flavinogenic yeasts. However, the mechanisms of such regulation are not known. We found that mutations causing riboflavin overproduction and iron hyperaccumulation (rib80, rib81 and hit1), as well as cobalt excess or iron deficiency all provoke oxidative stress in the Pichia guilliermondii yeast. Iron content in the cells, production both of riboflavin and malondialdehyde by P. guilliermondii wild type and hit1 mutant strains depend on a type of carbon source used in cultivation media. The data suggest that the regulation of riboflavin biosynthesis and iron assimilation in P. guilliermondii are linked with cellular oxidative state. PMID- 17910101 TI - Arbuscular mycorrhizal fungi in chronically petroleum-contaminated soils in Mexico and the effects of petroleum hydrocarbons on spore germination. AB - Arbuscular mycorrhizal fungi (AMF) have been hypothesized to enhance plant adaptation and growth in petroleum-contaminated soils. Nevertheless, neither AMF biodiversity under chronically petroleum-contaminated soils nor spore germination response to petroleum hydrocarbons has been well studied. Chronically petroleum contaminated rhizosphere soil and roots from Echinochloa polystachya, Citrus aurantifolia and C. aurantium were collected from Activo Cinco Presidentes, Tabasco, Mexico. Root colonization and spore abundance were evaluated. Additionally, rhizosphere soil samples were propagated using Sorghum vulgare L. as a plant trap under greenhouse conditions; subsequently, AMF-spores were identified. AMF-colonization ranged from 63 to 77% while spore number ranged from 715 to 912 in 100 g soil, suggesting that AMF tolerate the presence of petroleum hydrocarbons in the rhizosphere. From grass species, four AMF-morphospecies were identified: Glomus ambisporum, G. sinuosum (previously described as Sclerocystis sinuosum), Acaulospora laevis, and Ambispora gerdermanni. From citrus trees, four AMF-species were also identified: Scutellospora heterogama, G. ambisporum, Acaulospora scrobiculata, and G. citricola. In a second study, it was observed that spore germination and hyphal length of G. mosseae, G. ambisporum, and S. heterogama were significantly reduced by either volatile compounds of crude oil or increased concentrations of benzo[a ]pyrene or phenanthrene in water-agar. PMID- 17910102 TI - Characterization of the growth behavior of Leishmania tarentolae: a new expression system for recombinant proteins. AB - Biotechnological production of recombinant proteins for human therapy requires a cultivation of the host organism in a nutrient medium free of animal substances. Therefore, various nutrient media for the new expression system Leishmania tarentolae were developed and examined according to their cultivation conditions as static suspension culture and agitated culture. Investigations resulted in the development of a serum-free but hemin containing medium, based on yeast extract and buffer salts. Here we report that a high and stable specific growth rate of 0.103 h(-1) and a maximal cell density of 1 x 10(9) cells ml(-1) is obtained in an alternative medium, the YE-medium. For the newly developed medium, the successful expression of enhanced green fluorescent protein and the adaptation of the cultivation from the agitated culture to the bioreactor could be shown. Furthermore, an analytical method for detection of the essential, organic iron source hemin was established. The consumption of hemin was monitored because hemin is a potentially important process parameter for bioprocess control. With knowledge of these results, an improved expression system is available as an alternative to commonly used cell cultures for the production of recombinant proteins. PMID- 17910103 TI - Influence of soil compaction on microfungal community structure in two soil types in Bartin Province, Turkey. AB - Soil compaction negatively influences physical properties of soil (bulk density and pore space), and may consequently limit soil microfungi, which are significant for nutrient bioavailability. We measured microfungal community responses to compaction in a sandy loam and a clay loam soil at picnic sites. Soil bulk density increased significantly in the compacted samples. However, microfungal numbers and community composition were unrelated to changes in soil bulk density. With increases in bulk density from 1.22 to 1.37 g cm(-3) in the clay soil and from 1.38 to 1.54 g cm(-3) in the sandy loam soil, the total number of fungi declined or showed insignificant increases. In the compacted samples as well as the control sites, the most frequently occurring genera in the clay soil were Penicillium, Aspergillus, and Gliocladium. However, the most prominent feature occurring within the sandy loam soil was the exhibition of the greatest increase in the frequency of the Fusarium genus. When comparing compacted and control soils, fungal community composition corresponded more closely within each soil texture. The two microfungal soil communities, therefore, tolerated compaction. In contrast, a difference occurred in the fungal communities between the two soil textures. This is more likely due to the variability in the controlling factors of microfungal abundance and composition, such as soil characteristics, tree species, and competitive ability of fungal genera. PMID- 17910104 TI - Biodegradation of kerosene by Aspergillus ochraceus NCIM-1146. AB - The filamentous fungus Aspergillus ochraceus NCIM-1146 was found to degrade kerosene, when previously grown mycelium (96 h) was incubated in the broth containing kerosene. Higher levels of NADPH-DCIP reductase, aminopyrine N demethylase and kerosene biodegradation activities were found to be present after the growth in potato dextrose broth for 96 h, when compared with the activities at different time intervals during the growth phase. NADPH was the preferred cofactor for enzyme activity, which was inhibited by CO, indicating cytochrome P450 mediated reactions. A significant increase in all the enzyme activities was observed when mycelium incubated for 18 h in mineral salts medium, containing cholesterol, camphor, naphthalene, 1,2-dimethoxybenzene, phenobarbital, n-hexane, kerosene or saffola oil as inducers. Acetaldehyde produced by alcohol dehydrogenase could be used as an indicator for the kerosene biodegradation. PMID- 17910106 TI - Activity in vitro of twelve antibiotics against clinical isolates of extended spectrum beta-lactamase producing Escherichia coli. AB - Twelve beta-lactam and non-beta-lactam antibiotics were evaluated against 115 clinical isolates of extended-spectrum beta-lactamase-producing (ESBLs) Escherichia coli using a broth microdilution test in accordance with the CLSI guidelines. Susceptibility was 100% with imipenem, ertapenem and amikacin, 95.7% with piperacillin-tazobactam, 91.3% with cefoxitin, 87% with tobramycin, 81.7% with amoxicillin-clavulanate, 80% with cefepime, 67.8% with ceftazidime, 27.8% with ciprofloxacin, 27% with levofloxacin and 13% with ceftriaxone. Ertapenem was the antibiotic with the lowest minimum inhibitory concentrations (MICs) for all isolates. There were no clinically relevant differences in the activity of the antibiotics in the presence of CTX-M-9 or SHV enzymes. PMID- 17910105 TI - Production of L2 lipase by Bacillus sp. strain L2: nutritional and physical factors. AB - A thermophilic bacterium, Bacillus sp. strain L2 was isolated from a hot spring in Perak, Malaysia. An extracellular lipase activity was detected through plate and broth assays at 70 degrees C after 28 h of incubation. The L2 lipase production was growth dependent as revealed by a number of factors affecting the secretion of extracelullar lipase. As for nutritional factors, casamino acids, trehalose, Ca(2+) and Tween 60 were found to be more effective for lipase production. The optimum physical condition for L2 lipase production was obtained at 70 degrees C after 28 h of cultivation time, at pH 7.0, 150 rpm of agitation rate and 1% of starting inoculum size. The activity staining of crude L2 lipase revealed a clearing zone at 39 kDa. PMID- 17910107 TI - Alpha-amylase production by Bacillus subtilis CM3 in solid state fermentation using cassava fibrous residue. AB - In extraction of starch from cassava (Manihot esculenta Crantz), one of the major solid waste released is fibrous residues which constitute 15-20% by weight of the cassava chips/tuber processed. Production of alpha -amylase under solid state fermentation by Bacillus subtilis CM3 has been investigated using cassava fibrous residue. Response surface methodology (RSM) was used to evaluate the effect of the main variables, i.e. incubation period, initial medium pH, moisture holding capacity and temperature on enzyme production. A full factorial Central Composite Design (CCD) was applied to study these main factors that affected alpha -amylase production. The experimental results showed that the optimum incubation period, initial medium pH, moisture holding capacity and temperature were 6 days, 8.0, 70% and 50 degrees C, respectively. PMID- 17910108 TI - Projections of the costs associated with colorectal cancer care in the United States, 2000-2020. AB - Because of aging trends in the US, the number of prevalent colorectal cancer patients is expected to increase. We projected economic burden to the Medicare program and its beneficiaries through the year 2020. Burden was estimated for the initial phase of care, the period following diagnosis, the last year of life, and the continuing phase. Projected burden was evaluated with varying assumptions about incidence, survival, and costs of care. Estimated costs of care in 2000 in the initial, continuing, and last year of life phases of care were approximately $3.18 billion, $1.68 billion, and $2.63 billion, respectively. By the year 2020 under the 'fixed' current incidence, survival, and cost scenario, projected costs for the initial, continuing, and last year of life phases were $4.75 billion, $2.63 billion, and $4.05 billion. Under the current trends scenario (decreasing incidence, improving survival, and increasing costs), costs were $5.19 billion, $3.57 billion, and $5.27 billion. By the year 2020, estimated costs of colorectal cancer care among individuals aged 65 and older increased by 53% in the fixed scenario and by 89% in the current trends scenario. The future economic burden of colorectal cancer to the Medicare program and its beneficiaries in the US will be substantial. PMID- 17910109 TI - Use of instrumental variables in the presence of heterogeneity and self selection: an application to treatments of breast cancer patients. AB - Instrumental variable (IV) methods are widely used in the health economics literature to adjust for hidden selection biases in observational studies when estimating treatment effects. Less attention has been paid in the applied literature to the proper use of IVs if treatment effects are heterogeneous across subjects and individuals select treatments based on expected idiosyncratic gains or losses from treatments. In this paper we compare conventional IV analysis with alternative approaches that use IVs to estimate treatment effects in models with response heterogeneity and self-selection. Instead of interpreting IV estimates as the effect of treatment at an unknown margin of patients, we identify the marginal patients and we apply the method of local IVs to estimate the average treatment effect and the effect on the treated on 5-year direct costs of breast conserving surgery and radiation therapy compared with mastectomy in breast cancer patients. We use a sample from the Outcomes and Preferences in Older Women, Nationwide Survey which is designed to be representative of all female Medicare beneficiaries (aged 67 or older) with newly diagnosed breast cancer between 1992 and 1994. Our results reveal some of the advantages and limitations of conventional and alternative IV methods in estimating mean treatment effect parameters. PMID- 17910110 TI - A novel arterial pouch model of saccular aneurysm by concomitant elastase and collagenase digestion. AB - BACKGROUND: An ideal aneurysm model of cerebral aneurysm is of great importance for studying the pathogenesis of the lesion and testing new techniques for diagnosis and treatment. Several models have been created in rabbits and are now widely used in experimental studies; however, every model has certain intrinsic limitations. Here we report the development of a novel saccular aneurysm model in rabbits using an arterial pouch that is subject to in vitro pre-digestion with combined elastase and collagenase. METHODS: A segment of right common carotid artery (CCA) was dissected out and treated with elastase (60 U/ml, 20 min) followed by type I collagenase (1 mg/ml, 15 min) in vitro. The graft was anastomosed to an arterial arch built with the left CCA and the remaining right CCA, while the other end of the graft was ligated. The dimension and tissue structure of the pouch were analysed immediately, 2 or 8 weeks after operation. FINDINGS: Ten terminal aneurysms were produced. The gross morphology of the aneurysm resembles the human cerebral terminal aneurysms. We have observed the following pathological changes: (1) growth of the aneurysm (mean diameter increased from (2.0+/-0.1) to (3.2+/-0.3) mm at 2 weeks, P<0.001, n=7-10); (2) thinning of the aneurysmal wall (the mean wall thickness decreased to 44% at 2 weeks), which was accompanied by significant losses of elastic fibres, collagen and the cellular component; and (3) spontaneous rupture (3 out of 9, one aneurysm ruptured 24 h after operation with the other two at 2 and 4 weeks respectively). CONCLUSION: This rabbit arterial pouch model mimics human cerebral aneurysms in relation to morphology and histology. In particular, this model exhibited an increased tendency of spontaneous rupture. PMID- 17910111 TI - Deep lingual arterial chemoembolization of tongue carcinoma with microcapsuled anticancer drug. AB - OBJECTIVE: Microcapsule chemoembolism is a promising treatment of tumors. We describe a deep lingual arterial embolization of tongue carcinoma with microcapsuled carboplatinum. METHODS: Lingual artery cast specimens from cadavers were microscopically examined, and 78 patients with tongue cancer were recruited and treated with the deep lingual arterial embolization therapy. RESULTS: Microcapsule embolism occurred approximately at the fifth or sixth level of the deep lingual artery branches. The five-year survival rate was 88.5% (69 out of 78), and the ten-year survival rate 52.6% (41 out of 78). CONCLUSION: The deep lingual arterial embolization of tongue carcinoma with microcapsuled carboplatinum is an effective therapy to treat carcinoma in mid-margin or mid body of the tongue. PMID- 17910112 TI - Successful tubes treatment of esophageal fistula. AB - AIM: To discuss the merits of "tubes treatment" for esophageal fistula (EF). METHODS: A 66-year-old female who suffered from a bronchoesophageal and esophagothoratic fistula underwent a successful "three tubes treatment" (close chest drainage, negative pressure suction at the leak, and nasojejunal feeding tube), combination of antibiotics, antacid drugs and nutritional support. Another 55-year-old male patient developed an esophagopleural fistula (EPF) after esophageal carcinoma operation. He too was treated conservatively with the three tubes strategy as mentioned above towards a favorable outcome. RESULTS: The two patients recovered with the tubes treatment, felt well and became able to eat and drink, presenting no complaint. CONCLUSION: Tubes treatment is an effective basic way for EF. It may be an alternative treatment option. PMID- 17910113 TI - Lance-Adams syndrome: a report of two cases. AB - Chronic post-hypoxic myoclonus, also known as Lance-Adams syndrome (LAS), is a rare complication of successful cardiopulmanry resuscitation often accompanied by action myoclonus and cerebellar ataxia. It is seen in patients who have undergone a cardiorespiratory arrest, regained consciousness afterwards, and then developed myoclonus days or weeks after the event. Worldwide, 122 cases have been reported in the literature so far, including 1 case of Chinese. Here we report 2 Chinese LAS patients with detailed neuroimagings. Cranial single photon emission computed tomography (SPECT) of patient 1, a 52-year-old woman, showed a mild hypoperfusion in her left temporal lobe, whereas patient 2, a 54-year-old woman, manifested a mild bilateral decrease of glucose metabolism in the frontal lobes and a mild to moderate decrease of the N-acetyl aspartate (NAA) peak in the bilateral hippocampi by cranial [(18)F]-fluorodeoxyglucose positron emission tomographic (PET) scan and cranial magnetic resonance spectroscopy (MRS), respectively. We also review the literature on the neuroimaging, pathogenesis, and treatment of LAS. PMID- 17910114 TI - Pulmonary actinomycosis: a case undergoing resection through video-assisted thoracic surgery (VATS). AB - Actinomycosis is an uncommon disease, which is usually manifested as cervicofacial infection and related to poor oral hygiene or compromised immune function. Pulmonary actinomycosis is rare, but its diagnosis is changing due to its variable presentation and the similarity in appearance to other intrapulmonary diseases. Here we report an 80-year-old man with a solitary pulmonary nodule over the left upper lobe. Pulmonary neoplasm was highly suspected in this patient and thus resection of the mass was undertaken through video-assisted thoracic surgery (VATS). Histopathological examination demonstrated this patient had an Actinomyeces infection. While the application of VATS in patients with pulmonary actinomycosis has rarely been reported in literature, we conclude that VATS is valuable for the diagnosis and treatment of patients with undetermined pulmonary nodule(s). PMID- 17910115 TI - Influence of vermicomposting on solid wastes decomposition kinetics in soils. AB - The effect of vermicomposting on kinetic behavior of the products is not well recognized. An incubation study was conducted to investigate C mineralization kinetics of cow manure, sugarcane filter cake and their vermicomposts. Two different soils were treated with the four solid wastes at a rate of 0.5 g solid waste C per kg soil with three replications. Soils were incubated for 56 d. The CO(2)-C respired was monitored periodically and a first-order kinetic model was used to calculate the kinetic parameters of C mineralization. Results indicated that the percentage of C mineralized during the incubation period ranged from 31.9% to 41.8% and 55.9% to 73.4% in the calcareous and acidic soils, respectively. The potentially mineralizable C (C(0)) of the treated soils was lower in the solid waste composts compared to their starting materials. Overall, it can be concluded that decomposable fraction of solid wastes has decreased due to vermicomposting. PMID- 17910116 TI - Influence of nitrogen and sulfur fertilization on quality of canola (Brassica napus L.) under rainfed conditions. AB - Field experiments were conducted at Cereal Crops Research Institute, Pirsabak, Nowshera, Pakistan, during winter 2003-2004 and 2004-2005 to evaluate the effect of nitrogen and sulfur levels and methods of nitrogen application on canola (Brassica napus L. cv. Bulbul-98) under rainfed conditions. Four levels of S (0, 10, 20, and 30 kg/ha) and three levels of N (40, 60, and 80 kg/ha) and a control treatment with both nutrients at zero level were included in the experiments. Sulfur levels were applied at sowing while N levels were applied by three methods (100% soil application, 90% soil+10% foliar application, and 80% soil +20% foliar application). The experiments were laid out in randomized complete block (RCB) design having four replications. Oil content increased significantly up to 20 kg S/ha but further increase in S level did not enhance oil content. Glucosinolate content increased from 13.6 to 24.6 micromol/g as S rate was increased from 0 to 30 kg/ha. Protein content increased from 22.4% to 23.2% as S rate was increased from 0 to 20 kg/ha. Oil content responded negatively to the increasing N levels. The highest N level resulted in the highest values for protein (23.5%) and glucosinolate (19.9 micromol/g) contents. Methods of N application had no significant impact on any parameters under study. PMID- 17910117 TI - Characterizing and estimating rice brown spot disease severity using stepwise regression, principal component regression and partial least-square regression. AB - Detecting plant health conditions plays a key role in farm pest management and crop protection. In this study, measurement of hyperspectral leaf reflectance in rice crop (Oryzasativa L.) was conducted on groups of healthy and infected leaves by the fungus Bipolaris oryzae (Helminthosporium oryzae Breda. de Hann) through the wavelength range from 350 to 2,500 nm. The percentage of leaf surface lesions was estimated and defined as the disease severity. Statistical methods like multiple stepwise regression, principal component analysis and partial least square regression were utilized to calculate and estimate the disease severity of rice brown spot at the leaf level. Our results revealed that multiple stepwise linear regressions could efficiently estimate disease severity with three wavebands in seven steps. The root mean square errors (RMSEs) for training (n=210) and testing (n=53) dataset were 6.5% and 5.8%, respectively. Principal component analysis showed that the first principal component could explain approximately 80% of the variance of the original hyperspectral reflectance. The regression model with the first two principal components predicted a disease severity with RMSEs of 16.3% and 13.9% for the training and testing dataset, respectively. Partial least-square regression with seven extracted factors could most effectively predict disease severity compared with other statistical methods with RMSEs of 4.1% and 2.0% for the training and testing dataset, respectively. Our research demonstrates that it is feasible to estimate the disease severity of rice brown spot using hyperspectral reflectance data at the leaf level. PMID- 17910118 TI - Bioconversion of low quality lignocellulosic agricultural waste into edible protein by Pleurotus sajor-caju (Fr.) Singer. AB - Pleurotus sajor-caju (Fr.) Singer was cultivated on selected agro wastes viz. cotton stalks, groundnut haulms, soybean straw, pigeon pea stalks and leaves and wheat straw, alone or in combinations. Cotton stalks, pigeon pea stalks and wheat straw alone or in combination were found to be more suitable than groundnut haulms and soybean straw for the cultivation. Organic supplements such as groundnut oilseed cake, gram powder and rice bran not only affected growth parameters but also increased yields. Thus bioconversion of lignocellulosic biomass by P. sajor-caju offers a promising way to convert low quality biomass into an improved human food. PMID- 17910119 TI - Insecticidal activity of the medicinal plant, Alstonia boonei De Wild, against Sesamia calamistis Hampson. AB - The bioactivity of the aqueous extracts of the leaf and stem bark of the medicinal plant, Alstonia boonei De Wild (Apocyanaceae), against the pink stalk borer, Sesamia calamistis Hampson (Lepidoptera: Noctuidae) was studied in a laboratory bioassay. The extracts were incorporated into artificial diet at a rate of 0.0% (control), 1.0%, 2.5%, 5.0%, and 10.0% (w/w). Both extracts significantly (P<0.01) reduced larval survival and weight in a dose dependent manner. The concentrations that killed 50% of the larvae (LC(50)) for the stem bark extract were 2.8% and 2.1% at 10 and 20 DAI (days after introduction), respectively, while those for the leaves extract were 5.6% and 3.5%. The weights of the larvae also varied significantly (P<0.05) between the treatments in a dose dependent manner. We conclude that both leaf and stem bark extracts of A. boonei are toxic, used as growth inhibitors to S. calamistis larvae, and hold good promise for use as alternative crop protectants against S. calamistis. PMID- 17910120 TI - Stereoselective glucuronidation of carvedilol by Chinese liver microsomes. AB - OBJECTIVE: To study the stereoselective glucuronidation of carvedilol (CARV) by three Chinese liver microsomes. METHODS: The metabolites of CARV were identified by a hydrolysis reaction with beta-glucuronidase and HPLC-MS/MS. The enzyme kinetics for CARV enantiomers glucuronidation was determined by a reversed phase high pressure liquid chromatography (RP-HPLC) assay using (S)-propafenone as internal standard after precolumn derivatization with 2,3,4,6-tetra-O-acetyl-beta D-glucopyranosylisothiocyanate. RESULTS: Two CARV glucuronides were found in three Chinese liver microsomes incubated with CARV. The non-linear regression analysis showed that the values of K(m) and V(max) for (S)-CARV and (R)-CARV enantiomers were (118+/-44) micromol/L, (2 500+/-833) pmol/(min.mg protein) and (24+/-7) micromol/L, (953+/-399) pmol/(min.mg protein), respectively. CONCLUSION: These results suggested that there was a significant (P<0.05) stereoselective glucuronidation of CARV enantiomers in three Chinese liver microsomes, which might partly explain the enantioselective pharmacokinetics of CARV. PMID- 17910121 TI - Identification of novel catalytic features of endo-beta-1,4-glucanase produced by mulberry longicorn beetle Apriona germari. AB - Mulberry longicorn beetle, Apriona germari, has been reported to produce two endo beta-1,4-glucanases or AgEGases (accession Nos. Q6SS52 and Q5XQD1). AgEGase sequence contains catalytic motif (amino acid residues 37-48), which is the characteristic of family Glycohydrolase 45 and is identified as the substrate binding site. The application of bioinformatics approaches includes sequence analysis, structural modeling and inhibitor docking to relate the structure and function of AgEGases. We have dissected the sequence and structure of AgEGase catalytic motif and compared it with crystal structure of Humicola insolens endoglucanases V. The results show an involvement of sulfur containing amino acid residues in the active site of the enzyme. Cys residues and position of disulfide bonds are highly conserved between the two structures of endoglucanases of A. germari. Surface calculation of AgEGase structure in the absence of Cys residues reveals greater accessibility of the catalytic site to the substrate involving Asp42, a highly conserved residue. For the inhibition study, tannin-based structure was docked into the catalytic site of AgEGase using ArgusLab 4.0 and it resulted in a stable complex formation. It is suggested that the inhibition could occur through formation of a stable transition state analog-enzyme complex with the tannin-based inhibitor, as observed with other insect cellulases in our laboratory. PMID- 17910122 TI - Effects of polyurethane matrices on fungal tannase and gallic acid production under solid state culture. AB - The influence of the physical structure of polyurethane matrix as a support in a solid state culture in tannase production and gallic acid accumulation by Aspergillus niger Aa-20 was evaluated. Three different polyurethane matrices were used as the support: continuous, semi-discontinuous and discontinuous. The highest tannase production at 2479.59 U/L during the first 12 h of culture was obtained using the discontinuous matrix. The gallic acid was accumulated at 7.64 g/L at the discontinuous matrix. The results show that the discontinuous matrix of polyurethane is better for tannase production and gallic acid accumulation in a solid state culture bioprocess than the continuous and semi-discontinuous matrices. PMID- 17910123 TI - Disseminated cryptococcosis in a case of idiopathic CD 4 + lymphocytopenia. PMID- 17910126 TI - More articles needed from nurses in the trenches. PMID- 17910125 TI - Support for reducing use of restraint and seclusion. PMID- 17910127 TI - Losing valuable experience. PMID- 17910128 TI - Insuring independent midwives. PMID- 17910129 TI - Current world literature. Bladder cancer. PMID- 17910130 TI - [The water of the villages and the water of the countryside in Provence, 17th 20th centuries: the water problem in the area of Siagne]. PMID- 17910131 TI - Crime, suicide, and the anti-hero: "waltzing Matilda" in Australia. PMID- 17910132 TI - Dolls in Japan. PMID- 17910133 TI - [A survey of the International Symposium on Economic Development and Social Change]. PMID- 17910134 TI - [An outline of the history of the food supply]. PMID- 17910135 TI - [Violence against women and the new feminist challenges in Mexico]. PMID- 17910136 TI - Ethics self-assessment. PMID- 17910137 TI - Changes in the regulation of iodine crystals and chemical mixtures containing over 2.2 percent iodine. Final rule. AB - This rulemaking changes the regulation of the listed chemical iodine under the chemical regulatory provisions of the Controlled Substances Act (CSA). The Drug Enforcement Administration (DEA) believes that this action is necessary to remove deficiencies in the existing regulatory controls, which have been exploited by drug traffickers who divert iodine (in the form of iodine crystals and iodine tincture) for the illicit production of methamphetamine in clandestine drug laboratories. This rulemaking moves iodine from List II to List I; reduces the iodine threshold from 0.4 kilograms to zero kilograms; adds import and export regulatory controls; and controls chemical mixtures containing greater than 2.2 percent iodine. This rulemaking establishes regulatory controls that will apply to iodine crystals and iodine chemical mixtures that contain greater than 2.2 percent iodine. This regulation therefore controls iodine crystals and strong iodine tinctures/solutions (e.g., 7 percent iodine) that do not have common household uses and instead have limited application in livestock, horses, and for disinfection of equipment. Household products such as 2 percent iodine tincture/solution and household disinfectants containing iodine complexes will not be adversely impacted by this regulation. Additionally, the final rule exempts transactions of up to one-fluid-ounce (30 ml) of Lugol's Solution. Persons handling regulated iodine materials are required to register with DEA, are subject to the import/export notification requirements of the CSA, and are required to maintain records of all regulated transactions involving iodine regardless of size. PMID- 17910138 TI - Research ethics committees: preserving research integrity and the public trust. PMID- 17910139 TI - Patient satisfaction and quality of life among persons attending chronic disease clinics in South Trinidad, West Indies. AB - OBJECTIVE: Patient satisfaction and quality of life are increasingly being recognized as central elements in the monitoring and evaluation of healthcare. In this survey, the level of patient satisfaction and quality of life were investigated in regular attendees at public health chronic disease facilities in South Trinidad. METHOD: A random sample of 200 clients attending the three public chronic disease clinics during the period August 12, 2002 to December 31, 2002, completed self-administered questionnaires consisting of socio-demographic, quality of life (SF 12) and health service items. RESULTS: Participants had an average of four annual visits and 75% of them were 50 years and older. Approximately two-thirds of participants gave health and support staff a rating of good to excellent. Overall clinic experience was rated as poor to fair by 41.5%. Forty-five and a half per cent gave a rating of the explanations given by doctors and nurses about their illnesses. Fifty-three and a half per cent and 58% gave a poor to fair rating for the length of the waiting time and explanation offered when there was a significant delay in the starting times of clinics respectively. In regression analyses controlling for age, gender and number of illnesses, ratings of clinic experience and all categories of clinic staff were significantly associated with SF-12 mental and physical component summary scores. CONCLUSION: The findings suggest that in this population of regular clinic attendees, levels of client satisfaction and numbers of illnesses are associated with subjective quality of life. PMID- 17910140 TI - A knowledge, attitude and practices study of the issues of climate change/variability impacts and public health in Trinidad and Tobago, and St Kitts and Nevis. AB - OBJECTIVE: To determine the level of understanding of the issues of climate change (CC)/variability (CV) and public health by populations of St Kitts and Nevis (SKN) and Trinidad and Tobago (T&T) and to find whether respondents would be willing to incorporate these values into strategies for dengue fever (DF) prevention. DESIGN AND METHODS: Using a cluster sampling system, representative samples of the communities of SKN (227) and T&T (650) were surveyed for responses to a questionnaire document with questions on the impact of climate variability on health, the physical environment, respondents' willingness to utilize climate issues to predict and adapt to climate variability for DF prevention. Data were analyzed by Epi Info. RESULTS: Sixty-two per cent SKN and 55% T&T of respondents showed some understanding of the concept of climate change (CC) and distinguished this from climate variability (CV). With regard to causes of CC, 48% SKN and 50% T&T attributed CC to all of green houses gases, holes in the ozone layer burning of vegetation and vehicular exhaust gases. However some 39.3% SKN and 31% (T&T) did not answer this question. In response to ranking issues of life affected by CC/CV in both countries, respondents ranked them: health > water resources > agriculture > biodiversity > coastal degradation. The major health issues identified for SKN and T&T respondents were: food-borne diseases > water-borne diseases > heat stresses; vector-borne diseases were only ranked 4th and 5th for SKN and T&T respondents respectively. There was in both countries a significant proportion of respondents (p < 0.001) who reported wet season-related increase of DF cases as a CC/CV link. Respondents identified use of environmental sanitation (ES) at appropriate times as a method of choice of using CC/CV to prevent DF outbreaks. More than 82% in both countries saw the use of the CC/CV information for DF prevention by prediction and control as strategic but only 50-51% were inclined to become personally involved. Currently, only 50% SKN and 45% T&T respondents claimed current involvement in DF vector surveillance and control in the last two days. CONCLUSION: Despite the fact that knowledge and attitudes did not always coincide with practices of using ES for DF prevention, in both countries, even with CC/CV tools of prediction being available, it seems that respondents could be persuaded to use such strategies. There is a need for demonstration of the efficacy of CC/CV information and promotion of its usefulness for community involvement in DF and possibly other disease prevention. PMID- 17910141 TI - Effects of depression and anxiety on quality of life of patients with rheumatoid arthritis, knee osteoarthritis and fibromyalgia syndrome. AB - OBJECTIVE: To measure the effects of depression and anxiety on quality of life (QoL) in patients with rheumatoid arthritis (RA), knee osteoarthritis (OA) and fibromyalgia syndrome (FMS). METHODS: One hundred and fifty-four patients with RA, knee OA, and FMS who presented to the physical medicine and rehabilitation department were studied. For evaluation of the patients, Beck depression scale, Beck anxiety scale, and Short Form-36 were used. RESULTS: Twenty-two per cent of patients (n = 34) were diagnosed with of RA, 52.6% (n = 81) knee OA and 25.3% (n = 39) FMS. Except for the subscales, of physical and emotional role, there were statistically significant differences among diagnostic groups in the rest of the SF-36 subscales. In the physical functioning subscale, the highest score was obtained in the fibromyalgia group and the lowest in the RA group (p < 0.001). However, in the bodily pain subscale, the lowest score was recorded in the fibromyalgia group (p = 0.019). In all diagnostic groups, the scores of SF-36 subscales were significantly low in patients who scored above the threshold value of Beck depression scale (p < 0. 001). A strong negative correlation was detected between scores of Beck anxiety scale and the scores of all SF36 subscales in patients with RA and knee OA. On the other hand, in patients with FMS, anxiety scores correlated negatively with only physical and somatic function scores of SF 36. CONCLUSION: Quality of life is significantly low in patients with RA, knee OA and FMS, whose depression and/or anxiety scores are high. Therefore, these patients should be managed using a multidisciplinary approach including psychiatric support. PMID- 17910143 TI - Sensitivity and specificity of the electrocardiogram in predicting the presence of increased left ventricular mass index on the echocardiogram in Afro-Caribbean hypertensive patients. AB - Cardiovascular disease is emerging as the leading cause of death in the Caribbean region with hypertension along with diabetes mellitus representing the major causes. Left ventricular hypertrophy associated with hypertension results in a two to fourfold increase in cardiac morbidity and mortality. One hundred and eleven patients, 67% female, mean age 46 years with a mean of seven years since diagnosis, had resting blood pressure, electrocardiogram and sector-focused M mode echocardiogram performed The electrocardiograms were analyzed for left ventricular hypertrophy using Sokolow-Lyon, Cornell, Romhilts-Estes, 12 lead sum, QRS duration, 12 lead-QRS product and left ventricular strain pattern. The echocardiograms were analyzed for increased left ventricular mass using the formula of Devereux and Reichek indexed to height. The mean systolic blood pressure was 156 mmHg, mean diastolic blood pressure was 97 mmHg on treatment. At least one electrocardiographic criterion for left ventricular hypertrophy was seen in 47/111 (42%) patients and increased left ventricular mass index was seen in 55/111 (50%) patients. Sensitivity, specificity and positive predictive value of the electrocardiogram in predicting increased left ventricular mass index was best for Sokolow-Lyon (31%, 86%, 76%), Cornell (23%, 96%, 88%) and 12 lead-QRS product (30%, 86%, 72%). Sensitivity ranged from 3 to 31%, specificity from 80 to 96% and positive predictive value from 40 to 88%. The electrocardiogram is insensitive in detecting increased echocardiographic left ventricular mass index, as in patients from developed countries, and is less specific for the finding as in African Americans. PMID- 17910142 TI - Antinuclear antibodies and HLA class II alleles in Jamaican patients with systemic lupus erythematosus. AB - OBJECTIVE: The relationship between human leukocyte antigens class II (HLA) and antinuclear antibodies was investigated in Jamaican patients with Systemic Lupus Erythematosus (SLE). METHODS: Samples of blood of 82 patients with SLE and 75 healthy controls were tested for antinuclear antibodies using the fluorescent antinuclear antibody (FANA) test, counterimmunoelectrophoresis (CIEP) and the Crithidia luciliae immunofluorescence test (CL-IFT). A DNA-based HLA typing method was used to determine the frequencies of alleles of HLA-DRB1, DRB3, DRB4 and DRB5 in patients and healthy controls. RESULTS: The FANA test was positive in all of the sera from patients with SLE. Anti-dsDNA antibodies were present in 49% (40/82), anti-Sm/RNP 44% (36/82) and anti-Ro/La 43% (35/82) of the sera from SLE patients. The frequency of HLA-DR4 was significantly lower in SLE patients than in healthy controls (2/82, 2% vs 15/75, 20%; RR = 0.12; p = 0.0004; CP = 0.005) but no other HLA-DRB1 SLE associations were found. A positive HLA-DR3 anti-Ro/La antibody association was found in the patients with SLE (9/21, 43% vs 5/55, 9%; odds ratio (OR) = 7.5; CP = 0.01). In contrast, possession of HLA-DR6 was negatively associated with the absence of anti-dsDNA antibodies (9/32, 28% vs 27/44, 61%; OR = 0.2; CP = 0.05). CONCLUSION: The HLA-DR6 allele is associated with the absence of antinuclear antibodies and HLA-DR3 with the presence of anti Ro/La antibodies in Jamaican patients with SLE. However, these results and those of previous studies of Jamaican patients suggest that the HLA-DR3 association with the development of SLE reported in other populations might in fact reflect the association of HLA-DR3 with anti-Ro/La antibodies. Further investigations are needed to determine whether HLA-DRB antinuclear antibody associations define clinical subsets of SLE in Jamaican patients. PMID- 17910144 TI - Characteristics, treatment and short-term survival of patients with heart failure in a cardiology private practice in Jamaica. AB - BACKGROUND: Randomized clinical trials have demonstrated improvement in mortality with angiotensin converting enzyme inhibitors (ACEIs), beta blockers and aldosterone antagonists. The use of these lifesaving treatments remain inadequate. AIM: To determine the clinical features, aetiology, treatment and short-term survival of heart failure in a cardiology private practice in Jamaica. METHODS: This is a retrospective analysis of the medical records of 1055 consecutive patients presenting to a consultant cardiologist private practice between January 2002 and March 2003. Data were extracted from the records of the first 100 patients with heart failure. RESULTS: Most were over 65 years of age, female, never smoked cigarettes, overweight/obese and hypertensive (82%). The most commonly prescribed medications at one month were ACEIs (91%), beta blockers (88%) and loop diuretics (55%). The main aetiologies were hypertension (54%) and ischaemic heart disease (IHD) (26%). Ninety-one per cent were in sinus rhythm and 6% in atrial fibrillation. Forty-nine per cent had echocardiograms, of these 39% had ejection fractions (EF) > 40% and 27% had EF < or = 20%. The survival at one year was 81%. CONCLUSION: Hypertension was the major aetiology of heart failure followed by IHD. Medical treatment closely approached the recommended standards of major heart failure guidelines with high ACEI and beta blocker use comparable to recent heart failure trials. Short-term survival was very high. PMID- 17910145 TI - An evaluation of the intensive care unit resources and utilization in Trinidad. AB - OBJECTIVES: To evaluate resources and utilization of Intensive Care Units in Trinidad and Tobago. DESIGN AND METHODS: This was a prospective observational study to evaluate Intensive Care Units (ICU) of three public and two private hospitals in Trinidad with respect to their infrastructure, process of care and patient outcome. Structure of ICUs was assessed by interviews and personal observations. A Cost Block Model was used to determine the expenditure for ICUs. The process of ICU was assessed by Therapeutic Intervention Scoring System (TISS 28). For outcome evaluation, two prognostic scoring systems namely Simplified Acute Physiology Score (SAPS II) and Paediatric Index of Mortality-2 (PIM-2) were used RESULTS: The total number of ICU beds was 27. The overall bed occupancy was 66.2%. One hundred and eighteen patients consecutively admitted to ICU during a two-month period were enrolled for process and outcome evaluation. The overall median age of patients was 44 years [Interquartile range (IQR) 25, 59]. The mean cost per patient in the public hospitals was TT $64,746 compared to $77,000 in a private hospital. The average total daily TISS per patient was 27.01 +/- 5.4 (SD). The median length of stay was five days (IQR 2, 9). The overall predicted mortality was 32.9%, the observed mortality was 29.7% and thus the standardized mortality ratio (SMR) was 0.9. CONCLUSIONS: The overall bed availability in ICUs with respect to Trinidad and Tobago's population and case-mix is low compared to developed countries, although the process of ICU care is comparable. Outcome of patients was good in terms of risk-adjusted mortality. The study highlights the need to further increase bed-strength and optimize the resource utilization of ICUs in Trinidad and Tobago. PMID- 17910146 TI - Childhood sexual abuse among outpatients attending adult psychiatric outpatient clinics: a case-control study. AB - Only a few studies have focussed on the importance of routine investigation of childhood sexual abuse in outpatients attending adult psychiatric outpatient clinics. The aim of this study is to explore the association between having a history of childhood sexual abuse (CSA) and attending adult Psychiatric Outpatient Clinics in Trinidad. METHODS: This was a case-control study conducted in twelve psychiatric outpatient clinics located throughout Trinidad A questionnaire covering demographic, social, and sexual abuse components was administered by semi-structured interview to 566 participants, of whom 242 were cases, 239 were controls and 85 had incomplete questionnaires. The cases were 242 patients attending psychiatric outpatient clinics in Trinidad and the controls were 239 non-physician staffmembers at the clinics. Results were analyzed using the Stastistical Package for the Social Sciences (SPSS) version 10. RESULTS: Chi square analyses revealed several significant differences between the cases and control group. Sixty-three (26%) cases and 29 (12.1%) controls experienced CSA (p < 0.000). Twenty-five (39.7%) of the CSA cases had their experiences between the ages of 4 to 8 years and 13 (44.8%) of the CSA controls had their experiences between the ages of 9 to 12 (p < 0.01). Twenty-six (41.3%) of the cases and 3 (10.3%) of the controls had been abused at least 5 times (p < 0.000). Seventeen (58.6%) abused CSA controls reported having been sexually abused as a child only once. CSA with both force and manipulation was reported by 30 (47.6%) CSA cases while 6 (20.7%) CSA controls experienced CSA with force and manipulation (p < 0. 025). The abused CSA cases reported having a smaller social network of 2 persons compared to the abused CSA controls who had a social network of more than 4 persons (p < 0. 05). Of the 92 abused participants, 73.9% were women, and only 52.2% had told someone about the CSA. For the majority of CSA cases and CSA controls, the abuse involved one abuser. CONCLUSION: A positive correlation was established between earlier onset of CSA, repeated abuse (occurring more than 5 times), a limited social network in patients who had CSA and attending adult psychiatric outpatient clinics. Identifying CSA in psychiatric outpatients may lead to early intervention and aid patient management. PMID- 17910147 TI - Mammographic referral patterns for two breast imaging units in Jamaica. AB - OBJECTIVE: In countries that have instituted national mammographic screening programmes, mortality from breast cancer has decreased by as much as 63%. Although mortality rates from breast cancer in Jamaica are high, there is no national mammographic screening programme. In this context, opportunistic screening, which depends on contact between healthcare provider and patient, as well as self-referral become important. Therefore, the authors sought to determine the source of referrals for women who had mammography. SUBJECTS AND METHODS: The variables of age, indication for mammography, source of referral and referring physician area of specialty if applicable were extracted from the attendance records for all patients who had mammography at the breast imaging unit at the University Hospital of the West Indies (UHWI) and Radiology West (RadWest) in the year 2003. RESULTS: There were 779 bilateral mammograms done at UHWI of which 452 (58%) were screening and 1223 mammograms done at RadWest of which 657 (54%) were screening. The difference in proportion of self-referral between the two facilities was significantly different (p < 0. 001). Of the 452 screening mammograms performed at UHWI, 329 (73%) were self-referred, 31 (7%) were from primary care, 18 (4%) from gynaecologists and 17 (4%) from general surgeons. In contrast, of the 657 screening mammograms, at Radwest, 92 (14%) were self-referred, 323 (49%) were from primary care, 47 (7%) from gynaecologists and 37 (6%) from general surgeons. CONCLUSION: To increase the utilization and hence effectiveness of screening mammography, programmes targeting healthcare professionals, particularly gynaecologists and the public are needed. PMID- 17910150 TI - Changing the research culture at the section of psychiatry, the University of the West Indies, Mona. AB - OBJECTIVE: To compare the medical research output of the Section of Psychiatry, The University of the West Indies (UWI), Mona, before and after the implementation of strategies aimed at stimulating research. METHOD: Specific strategies such as weekly research and journal club meetings, with an emphasis on team activities and the establishment of bi-annual targets for submission of research papers were instituted in 2000. All research outputs from the Section of Psychiatry over the period 1995 to 2005 were identified from the Departmental Reports of the University of the West Indies and the published abstracts of the UWI Faculty of Medical Sciences and the Caribbean Health Research Council annual research conferences. A number of variables were extracted from each paper and comparisons made between the five-year period before and the five-year period after the implementation of the research enhancing strategies. Statistical analyses were performed with the Statistical Package for the Social Sciences (SPSS; version 11.5) and included chi-squared and Mann Whitney U tests. RESULTS: One-hundred and sixty-two items of research output were identified for the entire period under study. In the period after the implementation of the research enhancing strategies, there were significant increases in the total research output (p = 0.008) and refereed publications (p = 0.016). CONCLUSIONS: There were considerable increases in the overall research output of the department as well as in many sub-categories of output. These strategies are presented as a model to other departments seeking to augment their output of research. PMID- 17910148 TI - Towards research equity -- challenges of safety monitoring during clinical trials in resource-limited settings. PMID- 17910149 TI - Congenital toxoplasmosis in two health institutions in Trinidad. AB - Toxoplasmosis is the most widespread zoonosis and an important human disease particularly in children where it could cause visual and neurological impairment and mental retardation. This study was conducted to determine the prevalence of toxoplasmosis, especially congenital toxoplasmosis in patients at two health institutions in Trinidad A total of 504 cord blood samples of newborn babies were collected: 174 from a women's hospital and 330 from a general hospital. In order to elicit aternal and prenatal risk factors for toxoplasmosis, mothers of the newborns completed a questionnaire. Enzyme-immuno assay (EIA) was used to detect IgG and IgM to Toxoplasma gondii. Overall, of 504 serum samples tested, 220 (43.7%) were seropositive for IgG while the prevalence of congenital toxoplasmosis as reflected by IgM was 0.4%. The prevalence of IgG and IgM by health institutions was not significantly different (p > 0.05; chi-square). The prevalence of toxoplasmosis using IgG was highest in neonates of mothers who were of East Indian descent (54.1%), had four children (52.9%), kept cats in households (47.7%), practised outdoor gardening (50.8%), consumed raw meat (66.7%), had experienced miscarriage(s) (47.3%), stillbirths (66.7%), or who had eye problem(s) (52.9%) and mental retardation (50.0%). The study prevalence of congenital toxoplasmosis revealed a high seroprevalence oftoxoplasmosis in neonates but there was 0.4% serological evidence of congenital disease. It indicates a need for sensitization of the population and healthcare workers and for follow-up of infected children for clinical evidence of the disease. This would be necessary to fully appreciate the impact of toxoplasmosis in Trinidad and Tobago. The differences from comparison groups were however not statistically significant (p > 0.05; chi-square). PMID- 17910151 TI - The lungs in tuberous sclerosis complex. A case report. AB - Tuberous sclerosis complex manifests predominantly as a neurocutaneous disorder Lung involvement was considered rare. Lymphangioleiomyomatosis which occurs mainly in women of childbearing age is the major pulmonary disorder seen in tuberous sclerosis. Multifocal micronodular pneumocyte hyperplasia has also been described in tuberous sclerosis. The case of a 51-year old female diagnosed with tuberous sclerosis is described after she presented with progressive shortness of breath and was found to have interstitial lung disease. Tuberous sclerosis should be considered as a differential in patients with interstitial lung disease especially in association with cutaneous lesions. PMID- 17910152 TI - Insulinoma induced hypoglycaemia in a Jamaican patient. AB - Herein reported is the case of a young woman who had hyperinsulinaemic hypoglycaemia which was biochemically consistent with an insulinoma. Initial imaging was negative and definitive treatment was delayed until repeat imaging localized the tumour several years later. This case demonstrates the importance of clinical judgment and biochemical testing in the diagnosis of insulinoma despite negative imaging. PMID- 17910153 TI - Synchronous carcinoid tumour of the small intestine and appendix in the same patient. AB - Carcinoid tumours have been reported in a wide range of organs but most frequently involve the gastrointestinal tract. Many of these carcinoid tumours are associated with metachronous and synchronous lesions of another histological type. Primary carcinoid tumours of the different organ in the same patient is rare. In this paper, the authors present a case with synchronous carcinoid tumour of the small intestine and appendix in the same patient. PMID- 17910155 TI - Foreword: imagining a new era of neuroimaging, neuroethics, and neurolaw. PMID- 17910154 TI - Periodontal disease severity and systemic diseases prevalent in a Caribbean catchment area of patients. AB - OBJECTIVE: To describe associations between the prevalence of periodontal disease severity and co-existence of systemic disease(s) and a smoking habit amongst periodontal referrals in a Caribbean catchment area of patients. METHODS: A total of 100 patients completed a medical history questionnaire and were categorized for periodontal disease severity, using clinical and radiographic parameters for association with the prevalence of systemic diseases. RESULTS: Twenty-two per cent presented with moderate periodontal disease (M/F ratio: 1:2.7). 68% of patients examined presented with severe periodontal disease (M/F ratio: 1:1.35). Amongst patients of the same mean age of 48 years presenting with moderate or severe periodontal disease, there was a two-fold increase in the number of missing teeth, amongst patients with severe periodontal disease. In this category there was twice the proportion of smokers and twice the number of mobile teeth, compared with those with moderate periodontal disease. In addition, there was twice the prevalence of diabetics and three times the proportion of patients with combined systemic diseases amongst those with severe periodontal disease, compared with those presenting with moderate periodontal disease, who were predominantly hypertensive or had rheumatoid arthritis. A history of smoking, diabetes mellitus and a combined manifestation of systemic diseases appeared to be more prevalent amongst those with severe periodontal disease. All these findings were significant (p < 0.001). CONCLUSION: An association between severity of periodontal disease and co-existence of systemic diseases may have implications for a unified therapeutic strategy for health. PMID- 17910156 TI - Imaging the mind, minding the image: an historical introduction to brain imaging and the law. PMID- 17910157 TI - Imaging body structure and mapping brain function: a historical approach. PMID- 17910158 TI - MRIs and the perception of risk. PMID- 17910159 TI - Is a picture worth a thousand words? Neuroimaging in the courtroom. PMID- 17910160 TI - The brain and behavior: limitations in the legal use of functional magnetic resonance imaging. PMID- 17910161 TI - The regulation of MR neuroimaging research: disentangling the Gordian knot. PMID- 17910162 TI - FMRI and BF meet FRE: brain imaging and the federal rules of evidence. PMID- 17910163 TI - A brave new world of interrogation jurisprudence? PMID- 17910164 TI - Emerging neurotechnologies for lie detection and the fifth amendment. PMID- 17910165 TI - Neuroscience-based lie detection: the urgent need for regulation. PMID- 17910166 TI - Pain detection and the privacy of subjective experience. PMID- 17910168 TI - Interrogational neuroimaging in counterterrorism: a "no-brainer" or a human rights hazard? PMID- 17910167 TI - Punishment, freedom, and the culture of control: the case of brain imaging and the law. PMID- 17910169 TI - A double-edged sword: the role of neuroimaging in federal capital sentencing. PMID- 17910170 TI - How goals affect the organization and use of domain knowledge. AB - Expert specialists organize their knowledge around information related to their goals. In the experiments presented here, the relation between goal use and knowledge organization was investigated by manipulating participants' goals as they learned about a novel domain. Experiment 1 showed that goal use produces biases toward goal-related information in categorization and induction. Experiment 2 revealed that the bias toward goal relatedness is not absolute; participants use their knowledge flexibly, depending on the context of induction. Experiment 3 showed that using information in the absence of a meaningful goal does not produce significant goal-related biases. Altogether, the effects of goal use are evident across a number of tasks and may depend on goal meaningfulness and the coherence it provides to goal-related knowledge structures. PMID- 17910172 TI - Secondary-task effects on classification learning. AB - Probabilistic classification learning can be supported by implicit knowledge of cue-response associations. We investigated whether forming these associations depends on attention by assessing the effect of performing a secondary task on learning in the probabilistic classification task (PCT). Experiment I showed that concurrent task performance significantly interfered with performance of the PCT. Experiment 2 showed that this interference did not prevent learning from occurring. On the other hand, the secondary task did disrupt acquisition of explicit knowledge about cue-outcome associations. These results show that concurrent task performance can have different effects on implicit and explicit knowledge acquired within the same task and also underscore the importance of considering effects on learning and performance separately. PMID- 17910171 TI - Tracking mouse movement in feature inference: category labels are different from feature labels. AB - In this article, we examine the role of category labels in inductive inference. Some leading research has suggested that information about category membership works just like any other feature in categorical inductions, whereas other research has proposed that the influence of category membership on induction goes beyond that of other features. To investigate these claims further, we developed an online measure of judgments that is akin to eyetracking. The judgment results and the mouse-tracking data jointly support the view that category labels do affect inductive inferences in a way distinct from that for feature information. When arbitrary labels conveyed category membership information, participants viewed these labels more often and earlier in a trial, in comparison with cases in which the same labels conveyed non-membership information. Our results suggest that category membership information works like a guide for inference. An ecological rationale for this induction strategy is also discussed. PMID- 17910173 TI - The role of age and prior beliefs in contingency judgment. AB - This experiment investigated how prior beliefs affect young and older adults' ability to detect differences in objective contingency. Participants received new evidence that the objective contingency between two events was positive, negative, or zero when they believed that there was a positive or negative relationship between events, when they believed that the events were unrelated, and when they had no knowledge of the relationship between the events. They were then asked to estimate the objective contingency and recall the contingency evidence. Beliefs that events were or could be related improved young adults' contingency discrimination. Moreover, these beliefs did not produce biases in young adults' memory for the contingency evidence, but rather affected how they weighted this evidence at judgment. In contrast, these same beliefs did not improve older adults' contingency discrimination, but did produce biases in their memory for the evidence that were similar to those seen in their judgment. These findings are discussed in terms of age-related changes in working memory executive processes that impair older adults' ability to fully evaluate both belief-confirming and disconfirming contingency evidence and update their beliefs with this information. PMID- 17910174 TI - Cognitive complexity effects in perceptual classification are dissociable. AB - It has been proposed that a procedural-based classification system mediates the learning of information-integration categories, whereas a hypothesis-testing system mediates the learning of rule-based categories. Ashby, Ell and Waldron (2003) provided support for this claim by showing that a button switch introduced during classification transfer adversely affected information-integration but not rule-based performance. Nosofsky, Stanton and Zaki (2005) showed that increasing "cognitive complexity" can lead to button switch costs on rule-based performance. They argue that "cognitive complexity," and not the existence of separable classification systems, accounts for Ashby et al.'s empirical dissociation. The present study shows that experimental manipulations that increase "cognitive complexity" often have dissociable effects on information-integration and rule based classification that are predicted a priori from the processing characteristics associated with the procedural-based and hypothesis-testing systems. These results suggest that manipulations of "cognitive complexity" can be dissociated, suggesting that "cognitive complexity" in not a unitary construct that affects a single psychological process. PMID- 17910175 TI - Learning geographical information from hypothetical maps. AB - People show biases or distortions in their geographical judgments, such as mistakenly judging Rome to be south of Chicago (the Chicago-Rome illusion). These errors may derive from either perceptual heuristics or categorical organization. However, previous work on geographic knowledge has generally examined people's judgments of real-world locations for which learning history is unknown. This article reports experiments on the learning of hypothetical geographical spaces, in which participants acquired information in a fashion designed to control real world factors, such as variable travel experiences or stereotypes about other countries, as well as to mimic initial encounters with locations through reading or conventional school-based geography education. Five experiments combine to suggest that biases in judgment based on learning of this kind are different in key regards from those seen with real-world geography and may be based more on the use of perceptual heuristics than on categorical organization. PMID- 17910176 TI - Landmarks as beacons and associative cues: their role in route learning. AB - Relatively little is known about how people use the landmarks in their environment to learn routes. Landmarks are commonly regarded as associative cues- stimuli that enable recall of directional responses that lead closer to the navigator's goal. We contrast the function of landmark as associative cue with that of a beacon-a landmark near enough to a goal that moving toward it leads the navigator closer to his or her goal. In five experiments, participants learned a route through a simple desktop virtual environment. In the first three experiments, routes were learned better when their landmarks served as beacons than as associative cues. Two additional experiments showed that the acquired route knowledge depends on the function that landmarks serve during learning. Beacon-based route knowledge is less enduring and relatively less likely to involve knowledge of directions in the environment than is the route knowledge formed from landmarks that serve as associative cues. PMID- 17910177 TI - Long-term structural priming affects subsequent patterns of language production. AB - This article reports three experiments exploring how experience producing particular syntactic constructions affects the rates at which those constructions will be produced in the future. In the first part of each experiment, the participants' experience at producing the double object (DO) and prepositional object (PO) constructions was manipulated so that they produced a certain proportion of DO and PO constructions. Subsequent to the establishment of these biases, the participants were given the opportunity to produce either DO or PO constructions. The main findings show that (1) patterns of experience with the DO and PO constructions affected the base rates of production for the DO and PO constructions, but not the strength of structural priming observed between particular prime sentences and particular target sentences, and (2) patterns of experience with the DO and PO constructions affected the production of subsequent sentences even across changes in the nature of the language production task. The theoretical implications of these results are discussed. PMID- 17910178 TI - Cognate status and cross-script translation priming. AB - Greek-French bilinguals were tested in three masked priming experiments with Greek primes and French targets. Related primes were the translation equivalents of target words, morphologically related to targets, or phonologically related to targets. In Experiment 1, cognate translation equivalents (phonologically similar translations) showed facilitatory priming, relative to matched phonologically related primes, in conditions in which morphologically related primes showed no effect (50-msec prime exposure). Cross-language morphological priming emerged at longer prime exposure durations (66 msec), but cognate primes continued to generate more priming than did those in the morphological condition. In Experiments 2 and 3, the level of phonological overlap across translation equivalents was varied, and priming effects were measured against those for matched phonologically related primes and those in an unrelated prime condition. When measured against the unrelated baseline, cognate primes showed the typical advantage over noncognate primes. However, this cognate advantage disappeared when priming was measured against the phonologically related prime condition. The results are discussed in terms of how translation equivalents are represented in bilingual memory. PMID- 17910179 TI - Differences in semantic and translation priming across languages: the role of language direction and language dominance. AB - In the present study, we examined bilingual memory organization, using the priming paradigm. Many of the previous studies in which this experimental technique has been used in the bilingual domain appear to have had several differences in methodology that have caused there to be a lot of variation in the data reported. The aim of the present work was to create an experimental situation that was well constrained so that automatic processes could be observed. In Experiment 1, Spanish-English bilinguals participated in an unmasked semantic- and translation-priming study in which a lexical decision task was used. The results revealed significant translation-priming effects in both language directions and, unexpectedly, significant semantic priming in the L2-L1 direction only. In Experiment 2, we examined semantic- and translation-priming effects with a forward mask design. The results indicated that significant priming was obtained only for translation word pairs in both language directions. These results are discussed with regard to current models of bilingual memory representation. PMID- 17910180 TI - Syllable onsets are perceptual reading units. AB - Syllable onsets are defined as the initial consonant or consonant cluster in a syllable (e.g., BR in BREAD). In the present study, using a letter detection paradigm and French words, we tested whether syllable onsets are processed as units by the reading system. In Experiment 1, we replicated Gross, Treiman, and Inman's (2000) result of observing no difference between the detection latencies of letters embedded in a multi-letter syllable onset (e.g., c in ECLATER) relative to a single-letter syllable onset (e.g., C in ECARTER). In Experiment 2, participants took longer to detect the target letter when it was in the second position of a multi-letter onset (e.g., L in TABLIER) than when it was a single letter onset (e.g., L in ECOLIER). In Experiment 3, this position effect was replicated for graphemes. In Experiment 4, we tested and ruled out an alternative explanation of the unitization effect, in terms of lateral masking. These results indicate that syllable onsets are processed as perceptual units by the reading system and that the unitization effect is position dependent. PMID- 17910181 TI - Phonology as the source of syllable frequency effects in visual word recognition: evidence from French. AB - In order to investigate whether syllable frequency effects in visual word recognition can be attributed to phonologically or orthographically defined syllables, we designed one experiment that allowed six critical comparisons. Whereas only a weak effect was obtained when both orthographic and phonological syllable frequency were conjointly manipulated in Comparison 1, robust effects for phonological and null effects for orthographic syllable frequency were found in Comparisons 2 and 3. Comparisons 4 and 5 showed that the syllable frequency effect does not result from a confound with the frequency of letter or phoneme clusters at the beginning of words. The syllable frequency effect was shown to diminish with increasing word frequency in Comparison 6. These results suggest that visually presented polysyllabic words are parsed into phonologically defined syllables during visual word recognition. Materials and links may be accessed at www.psychonomic.org/archive. PMID- 17910182 TI - Feature and conjunction effects in recognition memory: toward specifying familiarity for compound words. AB - In three experiments, we evaluated potential sources of familiarity in the production of feature and conjunction errors with both word (Experiments 1 and 3) and nonword (Experiment 2) stimuli and related this work to various dual-process models of memory. The contributions of letter, syllable, lexical morpheme, and conceptual information were considered. Lexical morpheme information was consistently more potent than syllable information in leading to feature and conjunction errors across Experiments 1 and 2, and a word length explanation did not account for this consistent finding. In addition, there was no impact of conceptual information on these errors (Experiments 1-3). The results support a familiarity-based interpretation of feature and conjunction errors and a lexical morpheme basis for the familiarity in compound words. In order to be more comprehensive, memory models may need to account for a lexical morpheme source of familiarity. PMID- 17910183 TI - Can false memories be corrected by feedback in the DRM paradigm? AB - Normal processes of comprehension frequently yield false memories as an unwanted by-product. The simple paradigm now known as the Deese/Roediger-McDermott (DRM) paradigm takes advantage of this fact and has been used to reliably produce false memory for laboratory study. Among the findings from past research is the difficulty of preventing false memories in this paradigm. The purpose of the present experiments was to examine the effectiveness of feedback in correcting false memories. Two experiments were conducted, in which participants recalled DRM lists and either received feedback on their performance or did not. A subsequent recall test was administered to assess the effect of feedback. The results showed promising effects of feedback: Feedback enhanced both error correction and the propagation of correct recall. The data replicated other data of studies that have shown substantial error perseveration following feedback. These data also provide new information on the occurrence of errors following feedback. The results are discussed in terms of the activation-monitoring theory of false memory. PMID- 17910184 TI - Expectation of a final cumulative test enhances long-term retention. AB - In the present study, subjects studied lists of words across four experimental conditions: whether (or not) subjects received initial testing for these lists and whether (or not) they were made aware of an upcoming final free-recall test. Initial testing enhanced final-test performance; however, subjects benefited more from initial testing when they also knew they would need to remember the information for a later test. The data suggest that holding an expectation of the final test encouraged the continued processing of study materials following an initial test, affecting the accessibility of these materials at the time of final recall. The results clearly illustrate how an expectation of a cumulative test might influence long-term retention, which may have important implications for educational practice. PMID- 17910185 TI - How does delayed testing reduce effects of implicit memory: context infusion or cuing with context? AB - Studying a word implicitly activates related associates that affect its recall in the extralist cuing task. Recall is more likely when one of these associates becomes the test cue and when other associates of the studied word activate this associate. Context disruptions occurring between study and test reduce these effects, suggesting that implicitly activated memories are linked to context. The context infusion hypothesis assumes that context information spreads throughout a word's associative network during study, decreasing with distance from the studied word. Interactive cuing assumes that context is linked only to the studied word and that recall is based on retrieving information from the test cue and the context. The infusion hypothesis predicts that effects of disruptions will depend on link distance whereas interactive cuing predicts that distance will have no effect. The experiments evaluate these explanations by manipulating target-to-cue strength, associate-to-cue strength, and context disruption. Experiment 1 varies disruption by testing under the same or different conditions (room, modality, experimenter). Experiment 2 tests recall immediately or after 5 m, 10 m, or 20 m of multiplication. The results are inconsistent with context infusion and support the interactive cuing explanation. PMID- 17910186 TI - Source misattributions may increase the accuracy of source judgments. AB - Misattribution of remembered information from one source to another is commonly associated with false memories, but we demonstrate that it also may underlie memories that accord with past events. Participants imagined drawings of objects in four different locations. For each, a drawing of a similarly shaped object was seen in the same location, a different location, or not seen. When tested on memory for objects' origin (seen/imagined) and location, more false "seen" responses, but also more correct location responses, were given to imagined objects if a similar object had been seen, versus not seen, in the same location. We argue that misattribution of feature information (e.g., shape, location) from seen objects to similar imagined ones increased false memories of seeing objects but also increased correct location memories, provided the misattributed location matched the imagined objects' location. Thus, consistent with the source monitoring framework, imperfect source-attribution processes underlie false and true memories. PMID- 17910187 TI - The influence of strategic monitoring on the neural correlates of prospective memory. AB - The influence of strategic monitoring on the neural correlates of prospective memory was examined in two experiments. Strategic monitoring was manipulated by varying the context in which prospective cues occurred, with the cues requiring a prospective response in one task context and not the other. The N300, associated with cue detection, and the prospective positivity, associated with postretrieval processes, were elicited by prospective hits and were not elicited by prospective misses or ignore prospective cues. A parietal old-new effect was elicited by prospective misses and ignore prospective cues. Together these findings indicate that strategic monitoring may be necessary for cue detection and the recruitment of postretrieval processes, but not the recognition of a prospective cue as an old item. These findings serve to refine strategic monitoring and automatic associative accounts of prospective memory. PMID- 17910188 TI - Semantic and repetition priming effects for Deese/Roediger-McDermott (DRM) critical items and associates produced by DRM and unrelated study lists. AB - Two lexical decision experiments investigated priming for a critical item (CI, sleep) and its related yoked associate (YA, blanket) when one had been studied in a related Deese/Roediger-McDermott (DRM) list (Experiments 1 & 2) or a list of totally unrelated words (Experiment 2) and the other had been nonstudied. Semantic priming from the related DRM list occurred for nonstudied CIs (but not YAs) regardless of whether the CI received within-test priming from its studied related YA during the lexical decision task, though the effect in the absence of within-test priming averaged across experiments was only significant by a one tailed test. Also averaged across experiments, repetition priming occurred for both studied CIs and YAs when they had been studied in related DRM lists whether or not there was also within-test priming from a nonstudied related yoked pairmate, though individual effects within the two experiments were sometimes not significant. Repetition priming boosted semantic priming from related DRM lists less for CIs than for YAs, similar to the finding that memory discriminability is poorer for CIs than for YAs in episodic recognition. This smaller repetition priming boost for CIs than for YAs occurred to the same degree when the CIs or YAs were studied in an unrelated list. When nonstudied CIs and YAs were totally unrelated to all previously studied items and separated by 3-7 items in the lexical decision task, aYA produced a small 16 msec priming effect for its CI, averaged across both experiments. The implications of these results for the activation account of the DRM false-memory effect and for single-prime versus multiple-prime long-term semantic priming effects are discussed. The online addendum may be downloaded from www.psychonomic.org/archive. PMID- 17910189 TI - Metamemorial influences in recognition memory: pictorial encoding reduces conjunction errors. AB - Two experiments are presented that explore the role of the distinctiveness heuristic (e.g., Schacter, Israel, & Racine, 1999) on rates of conjunction errors as a function of encoding condition. The results of Experiment 1 demonstrate a reliable reduction of conjunction errors when participants study pictures relative to both reading words aloud and silently. Experiment 2 demonstrates that the nature of the pictures presented during the study phase is important for reducing conjunction errors. Collectively, the experiments demonstrate that participants can use the distinctiveness heuristic in addition to recall-to reject strategies to avoid conjunction errors. These findings add to a growing body of literature that suggests that participants are able to use expectations for memory to guide their recognition decisions. PMID- 17910190 TI - Unexpected costs of high working memory capacity following directed forgetting and contextual change manipulations. AB - Greater working memory capacity is usually associated with greater ability to maintain information in the face of interruptions. In two experiments, we found that some types of interruptions actually lead to greater forgetting among high span people than among low-span people. Specifically, an instruction designed to change mental context resulted in significant forgetting for high-span people but minimal forgetting among the low-span people. Intentional forgetting instructions also resulted in greater forgetting among higher working memory capacity participants than among lower working memory capacity participants. A candidate explanation called the intensified context shift hypothesis is proposed which suggests that high-span people are more context dependent than low-span people. PMID- 17910191 TI - Generation and mnemonic encoding induce a mirror effect in the DRM paradigm. AB - Encoding tasks that increase memory accuracy are appealing from both practical and theoretical perspectives. Within the Deese/Roediger-McDermott (DRM) paradigm, we found that generating list words from anagrams (relative to reading) produced a mirror effect: enhanced recognition of studied words coupled with a reduction in false recognition. Signal detection analyses suggest that the increase in correct recognition was due to enhanced item-specific encoding of the list words, whereas the reduction in false recognition was due to enhanced strategic monitoring at test (i.e., a distinctiveness heuristic), rather than to reduced relational encoding at study. Further support for a distinctiveness heuristic account was obtained using both "theme judgment" instructions and within-group conditions. In our final experiment, we replicated this mirror effect using a purely mnemonic (self-referential) encoding task, showing that extra perceptual cues are not necessary to induce participants to adopt a successful memory improvement strategy at test. PMID- 17910192 TI - Output order in immediate serial recall. AB - In two experiments, we examined the effect of output order in immediate serial recall (ISR). In Experiment 1, three groups of participants saw lists of eight words and wrote down the words in the rows corresponding to their serial positions in an eight-row response grid. One group was precued to respond in forward order, a second group was precued to respond in any order, and a third group was postcued for response order. There were significant effects of output order, but not of cue type. Relative to the forward output order, the free output order led to enhanced recency and diminished primacy, with superior performance for words output early in recall. These results were replicated in Experiment 2 using six-item lists, which further suggests that output order plays an important role in the primacy effect in ISR and that the recency items are most highly accessible at recall. PMID- 17910194 TI - Individual differences in phonological learning and verbal STM span. AB - A relationship between phonological short-term memory tasks (e.g., nonword repetition, digit span) and vocabulary learning in both experimental and real life conditions has been reported in numerous studies. A mechanism that would explain this correlation is, however, not known. The present study explores the possibility that it is the quality of phonological representations that affects both short-term recall and long-term learning of novel wordlike items. In Experiment 1, groups with relatively good and poor span for pseudowords were established. The good group was found to perform better at explicit memory tasks tapping the incidental learning of a limited stimulus pool used in an auditory immediate serial pseudoword recall task. In Experiment 2, the results of Experiment 1 were replicated when experience of correct recall was controlled. In Experiment 3, the immediate recall performance of the good group was found to benefit more than that of the poor group from syllable repetition within stimulus pools. It is concluded that the efficiency of a process that creates phonological representations is related both to short-term capacity for verbal items, and to long-term phonological learning of the structure of novel phonological items. PMID- 17910193 TI - Working memory contributions to relative clause attachment processing: a hierarchical linear modeling analysis. AB - An eye-movement-monitoring experiment tested readers' responses to sentences containing relative clauses that could be attached to one or both of two preceding nouns. Previous experiments with such sentences have indicated that globally ambiguous relative clauses are processed more quickly than are determinately attached relative clauses. Central to the present research, a recent study (Swets, Desmet, Hambrick, & Ferreira, 2007) showed that offline preferences for such sentences differ as a function of working memory capacity. Specifically, both English and Dutch participants' preference for the second of two nouns as the host for the relative clause increased as their working memory capacity increased. In the present study, readers' working memory capacity was measured, and eye movements were monitored. Hierarchical linear modeling was used to determine whether working memory capacity moderated readers' online processing performance. The modeling indicated that determinately attached sentences were harder to process than globally ambiguous sentences, that working memory did not affect processing of the relative clause itself, but that working memory did moderate how easy it was to integrate the relative clause with the preceding sentence context. Specifically, in contrast with the offline results from Swets and colleagues' study, readers with higher working memory capacity were more likely to prefer the first noun over the second noun as the host for the relative clause. PMID- 17910195 TI - The validity of "conceptual span" as a measure of working memory capacity. AB - Three experiments tested whether a modified version of the Clustered Conceptual Span task (H. J. Haarmann, E. J. Davelaar, & M. Usher, 2003), which ostensibly requires active maintenance of semantic representations, predicted individual differences in higher-order cognitive abilities better than short-term memory (STM) span tasks or nonsemantic versions of the "Conceptual" task did. Nonsemantic Conceptual tasks presented short word lists clustered by color, first letter, or initial vowel sound, and cued subjects to recall only 1 of 3 clusters from each list; the Semantic task clustered words by taxonomic category. The Semantic Conceptual task generally failed to predict incremental variance in either verbal abilities or general fluid intelligence beyond the other Conceptual tasks or STM span tasks. Although the Semantic task showed a stronger relation to working memory span tasks than did the nonsemantic tasks (Experiment 3), that stronger relation did not translate into strong prediction of cognitive individual differences. PMID- 17910196 TI - Is there an internal association of numbers to hands? The task set influences the nature of the SNARC effect. AB - The SNARC effect refers to the association of smaller numbers with the left and of larger numbers with the right side of extracorporal space (Dehaene, Bossini, & Giraux, 1993). We tested the assumption that, in addition to these associations, numbers are also related to participants' hands. We report two experiments with vertically arranged buttons in which the nature of the SNARC effect depended on whether the task set was button or hand related: In the first case, a vertical location-related SNARC effect occurred, whereas in the second a hand-related SNARC effect was found. Our third experiment confirmed that space-related number representations dominate the SNARC effect when the buttons are arranged horizontally. We concluded that both effector- and space-related number representations can influence and modify the SNARC effect. PMID- 17910197 TI - Paying attention to binding: further studies assessing the role of reduced attentional resources in the associative deficit of older adults. AB - The present experiments investigated whether the observed associative deficit in older adults' episodic memory is mediated by a reduction of attentional resources. Using a dual-task procedure, younger and older participants studied lists of word pairs either under full attention or while performing a concurrent task. Both experiments showed that dividing attention did not cause a greater impairment to memory for associations than to memory for items in either age group. Furthermore, an analysis of concurrent task performance revealed that older adults' attentional costs for both learning and binding items were not larger than for learning items alone, relative to younger adults. These data provide support for a multicausal interpretation of older adults' memory deficits in which common, depleted attentional resources may be a mechanism that reduces memory for components of an episode in both older and younger adults under divided attention at encoding. In addition, older adults have a unique deficit in memory for the associations between the components, which does not seem to be resource dependent. PMID- 17910199 TI - Psychopharmacology. PMID- 17910198 TI - The costs and benefits of cross-task priming. AB - Two lines of research on cross-task priming yield opposite results. Research on repetition priming observed positive priming, whereas research on the role of priming in task-switching observed negative effects. We combined the two types of design. In the transfer phase of our paradigm, subjects performed task B either as a pure block (BBB) or as a switch block (ABAB). We presented items which were either unprimed or primed by prior presentation during a preceding priming phase performed on task A. Amongst others, the priming effect is determined by two factors: First, the more operation time the system needs during the probe event, the higher the likelihood to obtain priming. Protracting operation time by reducing stimulus quality favors positive priming, whereas providing more operation time by making subjects switch between tasks favors negative priming. Second, the strength of the memory trace of the prime event determines whether that trace can possibly yield negative priming, in that only strong traces can be retrieved together with the associated task/response. PMID- 17910200 TI - A 9-year-old girl with severe anemia. PMID- 17910201 TI - A 5-year-old girl with developmental delay. PMID- 17910202 TI - Promoting access to quality psychopharmacology services for youths. PMID- 17910203 TI - Medical management of pediatric mood disorders. PMID- 17910205 TI - Pharmacologic treatment of autism and related disorders. PMID- 17910204 TI - Psychopharmacologic treatment of attention-deficit/hyperactivity disorder and disruptive behavior disorders. PMID- 17910206 TI - Pharmacotherapy for anxiety disorders in children and adolescents: an evidence based medicine review. PMID- 17910207 TI - Nutritional aspects of child and adolescent psychopharmacology. PMID- 17910208 TI - CMS releases proposed 2008 Medicare physician fee schedule. PMID- 17910209 TI - Compassionate cruelty. PMID- 17910210 TI - The evolution of the multispecialty group. How specialty mix changes with group size. PMID- 17910211 TI - The good, the bad and the equitable. Patient safety and the just culture. PMID- 17910212 TI - Help in the hinterlands. A change in immigration policy opens door for foreign born physicians. PMID- 17910213 TI - How can practice administrators make themselves, their practices better? Atul Gawande, MD, MPH, on the science of performance. PMID- 17910214 TI - Navigating your course in medical practice administration. What MGMA member research says about your career. AB - Plotting your career as a medical practice management professional can be a lot like charting the course of a small boat on open waters: The environment may be glassy-smooth, choppy and rife with undercurrents, or even as tumultuous as a hurricane. But you can learn from the experiences of others who've navigated the same seas. The MGMA Center for Research studied the career paths of medical practice administrators, focusing on the positions held during a career, the time spent in each and the reasons for changing jobs. The data and administrators' comments provide a compelling portrait of your profession. PMID- 17910215 TI - Developing the next generation of leaders. AB - Linking the elements of management development to organizational culture and business goals is essential for a successful leadership program. Establishing such a program in your practice is not enough, however--an organization must also evaluate its effect on performance. For organizations that embrace and institute training and leadership development, the financial payoffs can be substantial. PMID- 17910216 TI - You can be a wizard of IDS. Is your practice ready to form an integrated delivery system? AB - To combat declining reimbursement and strengthen their positions in a highly competitive provider environment, leaders of large medical groups may consider forming an integrate delivery system (IDS) to gain a business advantage. Managing the array of organizational, operational, financial and human relationships affected by an IDS requires careful planning, good timing and a bit of luck. This article, the first of a two-part series, examines what questions physician practice leaders should ask--and answer--before formiing an IDS. PMID- 17910217 TI - Clinical assessment of postoperative sensitivity in posterior composite restorations. AB - This incidence of postoperative sensitivity was evaluated in resin-based posterior restorations. Two hundred and ninety-two direct restorations were evaluated in premolars and molars. A total of 143 Class I and 149 Class II restorations (MO/OD and MOD) were placed in patients ranging in age from 30 to 50 years. After the cavity preparations were completed, a rubber dam was placed, and the preparations were restored using a total-etch system (Prime & Bond NT) and a resin-based restorative material (TPH Spectrum). The patients were contacted after 24 hours and 7, 30 and 90 days postoperatively and questioned regarding the presence of sensitivity and the stimuli that triggered that sensitivity. The Chi square and Fisher's Exact Test were used for statistical analysis. Evaluation at 24 hours after restorative treatment revealed statistically significant differences among the types of cavity preparations restored and the occurrence of postoperative sensitivity (p = 0.0003), with a higher frequency of sensitivity in Class II MOD restorations (26%), followed by Class II MO/DO (15%) and Class I restorations (5%). At 7, 30 and 90 days after restorative treatment, there was a decrease in the occurrence of sensitivity for all groups. The percentage of sensitivity among the groups was not significantly different. This study shows that the occurrence of sensitivity is correlated with the complexity of the restoration. PMID- 17910218 TI - Clinical evaluation of 15% carbamide peroxide on the surface microhardness and shear bond strength of human enamel. AB - PURPOSE: This clinical evaluation compared a neutral sodium fluoridated whitening product to a neutral non-fluoridated whitening product in terms their effects on human enamel surface microhardness (SMH) and human enamel/resin composite shear bond strength (SBS) following various treatment times. MATERIALS AND METHODS: Subjects were evaluated for enamel SMH and enamel/resin SBS following 15% carbamide peroxide (CP) with and without potassium nitrate and fluoride (PF). Twenty subjects (80 first or second premolars), who were treatment-planned for premolar extraction due to orthodontic therapy, were allocated into two groups, A and B. Group A received 15% CP, while Group B received 15% CP with PF. Each patient had a control tooth, a 14-day treatment + 14-day recovery tooth, a 14-day treatment + no recovery tooth and a 4-day + no recovery tooth. Each tooth was further divided into two testing surfaces; the facial surface was used for SMH, while the lingual surface was used for SBS. RESULTS: The results of this study determined that there was no statistically significant difference between the effects of the two products on SMH and enamel/resin SBS. Additionally, there was no statistically significant difference between the treatment specimens compared to the controls in terms of SMH. However, there was a significant difference between the treatment groups compared to the controls in terms of enamel/resin SBS. CONCLUSION: Within the limitations of this clinical study, 15% CP with and without PF does not seem to alter the SMH of human enamel. However, 15% CP with and without PF significantly reduced enamel/resin SBS immediately following tooth whitening therapy, up to 14 days post-treatment. PMID- 17910219 TI - Direct resin composite restorations in vital versus root-filled posterior teeth: a controlled comparative long-term follow-up. AB - This study compared direct composite restorations on vital versus endodontically treated posterior teeth, six to eight years after placement. All clinical procedures were performed using standardized materials and protocols. The patients were identified with the aid of a computerized billing system followed by a chart search. With each patient, the prime investigator matched a randomly chosen endodontically-treated tooth (test tooth) with a vital counterpart (control tooth) with a direct composite restoration. Of the 84 patients who fulfilled the inclusion criteria, 44 were available for recall. Two calibrated observers, blinded to the tooth vitality status, evaluated the fillings by applying the modified USPHS criteria. The outcome was dichotomized to revision indicated/recommended vs revision not indicated. Odds Ratios (OR) and corresponding 95% Confidence Intervals (95% CI) were calculated. Residual confounding was corrected using a multivariable regression analysis with the remaining known confounders filling size, tooth type and age of the fillings in months as independent variables. The crude association between the presence/absence of root canal treatment and the need for revision (OR, 95% CI) was 2.12 (1.02-4.38; p = 0.04). Correcting for all potential confounders in the multivariable analysis changed this association marginally to an OR of 1.98 (0.90 4.38; p = 0.09). PMID- 17910220 TI - The effect of light curing source on the residual yellowing of resin composites. AB - PURPOSE: This study evaluated the amount of residual yellow in cured resin composites when polymerizing with either a quartz-tungsten-halogen (QTH) or blue light-emitting diode (LED). MATERIAL AND METHODS: Twelve shades (bleaching to conventional shades) of microfill, hybrid and microhybrid resin composite specimens (n = 10) were polymerized with both light types. All the materials contained only camphorquinone as the photoinitiator. After exposure, the specimens were stored in the dark for 24 hours. Then, the specimen color parameters were recorded (L*, a*, b* and C*(ab)) and color differences (deltaE*(ab)) were determined by examining for changes among the test combinations. Group comparisons were examined using ANOVA and the Tukey-Kramer post-hoc test, and pairwise comparisons were made using the Student's t-tests at a pre-set alpha of 0.05. RESULTS: When a significant difference in the shade of yellow was noted, the QTH light produced a greater yellow tinge than most comparisons using the LED. The potential for producing more residual yellowing could not be anticipated with respect to composite filler classification or shade, as this effect may be more dependent on individual product composition. The extent to which residual yellowing differences were noted between light curing units fell within levels considered detectable by the human eye (deltaE > 2.0). CONCLUSION: The selection of light curing unit to polymerize resin-based restorative materials can have a significant influence on the amount of residual yellow present, with the QTH light tending to leave more yellow than an LED unit. PMID- 17910221 TI - The effect of desensitizing treatments on the bond strength of resin composite to dentin mediated by a self-etching primer. AB - This study evaluated the bond strength of resin composite to dentin, mediated by a self-etching adhesive, following the application of various dentin desensitizing treatments and artificial saliva storage. The buccal cervical areas of 24 extracted human third molars were ground flat to expose cervical dentin. The dentin surfaces were polished with 1200-grit SiC paper, then the teeth were randomly assigned to six groups, five desensitizing treatments and one control: Group I-VivaSens; Group II-Fluor Protector; Group III-Isodan; Group IV-Futura Bond NR; Group V-Nd:YAG laser and Group VI-Control (without application of a desensitizing agent). After applying the desensitizing treatments and storing the molars in artificial saliva for 14 days at 37 degrees C, Futura Bond NR was used to bond resin composite to dentin. TPH composite build-ups were constructed incrementally to a height of 5 mm. The teeth were sectioned to obtain bonded slices of 0.7 mm thick specimens containing the resin-composite joint. The specimens were then trimmed into an hourglass shape and subsequently subjected to microtensile testing at a crosshead speed of 1 mm/minute. The data were analyzed using the Kruskal-Wallis analysis and multiple comparisons test. The control (Group VI) and Futura Bond NR self-etching treatment (Group IV) group yielded statistically significant higher bond strength values than the other desensitizing treatment groups tested (p < 0.005). While pretreatment of dentin surfaces with desensitizing agents (Fluor Protector, VivaSens and Isodan) and laser (Nd:YAG) reduced the bond strength values of the resin composite, higher bond strengths were achieved using a self-etching adhesive (Futura Bond NR) as a desensitizing agent. PMID- 17910223 TI - Effect of elastic cavity wall and occlusal loading on microleakage and dentin bond strength. AB - OBJECTIVE: This study evaluated the effect of an unfilled-adhesive resin (Adper Single Bond) and a filled-adhesive resin (Adper Single Bond 2) with and without a low viscosity resin (Filtek Flow) as an elastic cavity wall on marginal leakage and dentin microtensile bond strength in Class V composite restorations under unloaded and loaded conditions. METHODS: V-shaped cavities were prepared on the buccal surfaces of 56 premolars lined with unfilled (Groups 1 and 3) or filled (Groups 2 and 4) adhesives with (Groups 3 and 4) and without (Groups 1 and 2) a low viscosity resin and restored with a resin composite. The restored teeth in each group were divided into two sub-groups for unloaded and loaded conditions with 50N loading force for 250,000 cycles parallel to the long-axis of the tooth. Five specimens from each group were cut bucco-lingually 0.7 mm thick and subjected to a dye leakage test for four hours using 2% methylene blue dye. The tested specimens were then trimmed into dumbbell shapes at the gingival margin and subjected to microtensile testing. The remaining two specimens were cut, embedded and observed for resin/dentin interfaces under a scanning electron microscope. RESULTS: For the microleakage test, there were no significant differences in microleakage among the groups on both the enamel and dentin margin. No statistically significant differences were found between microleakage of the loaded and unloaded groups on enamel margins for all materials. There were statistically significant differences between microleakage of the loaded and unloaded groups on the dentin margin for Groups 3 and 4. For the microtensile test, the significant difference was found between Groups 1 and 4 for the unloaded groups. For the loaded groups, there were no significant differences between Groups 1 and 2 and Groups 3 and 4. There were no statistically significant differences in microtensile bond strength between the loaded and unloaded groups except for Group 2. CONCLUSION: The application of filled adhesive or low viscosity resin had no influence on marginal leakage at both the enamel and dentin margin but it had an influence on the microtensile bond strength to dentin of Class V restorations. Occlusal loading significantly increased the degree of marginal leakage at the dentin margin when low viscosity was applied in combination with either unfilled or filled-adhesives, but it decreased dentin bond strength in the group treated with only filled adhesive. PMID- 17910222 TI - The influence of internal surface treatments on tensile bond strength for two ceramic systems. AB - STATEMENT OF THE PROBLEM: The ceramic composition and surface microstructure of all-ceramic restorations are important components of an effective bonding substrate. Hydrofluoric acid and sandblasting are well-known procedures for surface treatment; however, surface treatment for high alumina-containing and lithium disilicate ceramics have not been fully investigated. PURPOSE: This in vitro study evaluated the tensile bond strength of resin cement to two types of ceramic systems with different surface treatments. METHODS AND MATERIALS: Thirty specimens of each ceramic system were made according to the manufacturer's instructions and embedded in polyester resin. Specimens of In-Ceram Alumina [I] and IPS Empress 2 [E] were distributed to three groups with differing surface treatments (n = 10): sandblasting with 50 microm aluminum oxide (APA); sandblasting with 110 microm aluminum oxide modified with silica particles (ROCATEC System-RS); a combination of sandblasting with APA and 10% hydrofluoric acid etching (HA) for two minutes on In-Ceram and for 20 seconds for IPS Empress 2. After the respective surface treatments, all the specimens were silanated, and Rely-X resin cement was injected onto the ceramic surface and light polymerized. The specimens were stored in distilled water at 37 degrees C for 24 hours and thermally cycled 1,100 times (5 degrees C/55 degrees C). The tensile bond strength test was performed in a universal testing machine at a 0.5 mm/minute crosshead speed. RESULTS: The mean bond strength values (MPa) for IPS Empress 2 were 12.01 +/- 5.93 (EAPA), 10.34 +/- 1.77 (ERS) and 14.49 +/- 3.04 (EHA). The mean bond strength values for In-Ceram Alumina were 9.87 +/- 2.40 (IAPA) and 20.40 +/- 6.27 (IRS). All In-Ceram specimens treated with 10% hydrofluoric acid failed during thermal cycling. CONCLUSION: The Rocatec system was the most effective surface treatment for In-Ceram Alumina ceramics; whereas, the combination of aluminum oxide sandblasting and hydrofluoric acid etching for 20 seconds worked more effectively for Empress 2 ceramics. PMID- 17910225 TI - Sealing of minimally invasive Class II fillings (slot) using an adhesive patch: sealant margin extension for prevention. AB - A laboratory study was performed to assess the potential of an adhesive patch to seal small, unbeveled, Class II, box-only (slot) composite fillings. After minimal access cavity preparation with an 80 pm diamond bur, 40 box-only Class II cavities were prepared mesially and distally in 20 extracted human molars using a u-shaped PCS insert (EMS). One cavity per tooth was adhesively filled with a hybrid composite material in one increment. A patch, acting as an adhesive matrice, was applied to the other cavity of each tooth to seal the restoration. The margin of the patch was located in areas easily accessible to oral hygiene measures and self-cleaning. All the teeth were subjected to thermo-mechanical stress in a computer-controlled masticator device. In 10 teeth, caries was induced in a microbial-based artificial mouth model and quantitatively determined by confocal laser scanning microscopy. Microleakage was assessed in the other 10 teeth in two planar sections after immersion in 0.5% basic fuchsin solution. The results showed no demineralization at the filling margins protected with the patch. Microleakage was observed in one sample only and was limited to the enamel. In contrast, the margins of fillings without the patch application showed a mean demineralization depth of 146 +/- 42 microm and dye penetration into the dentin in five sections. This innovative approach to sealing restorative margins with an adhesive patch results in less leakage and filling margin demineralization and merits further investigation. PMID- 17910224 TI - Effect of carbamide peroxide treatments on the metal-ion release and microstructure of different dental amalgams. AB - OBJECTIVES: A variety of methods have been used to model the effects of bleaching agents on tooth tissue and dental materials. Although several studies have evaluated the effects of bleaching agents on restorative materials, little attention has been directed to metal ion release from dental amalgam following exposure to bleaching agents. This in vitro study investigated the effects of two carbamide-peroxide-based bleaching gels on metal ion release from different dental amalgams. METHODS: Thirteen discs (10x2 mm) prepared from tgloy (non-gamma 2, admix amalgam), Septalloy NG 50 (single composition, non-spherical, non-gamma 2, ternary amalgam), Dispersalloy (dispersed phase, admix amalgam) and Permite C (high-copper, non-gamma 2, admix alloy), according to the manufacturers' instructions and polished after 24 hours, were aged for seven days at 37 degrees C in air. The discs were then immersed in 10 ml of distilled water for 24 hours at 37 degrees C. The amount of metal ion (Hg, Ag, Sn and Cu) release was determined using inductively coupled plasma mass spectrometry. The discs were treated with either 16% or 30% carbamide peroxide gel and ion release analysis was repeated. The specimens were also evaluated for surface changes using scanning electron microscopy (SEM). The data were analyzed using the MannWhitney U, Wilcoxon Signed Ranks and Kruskal Wallis tests. RESULTS: Statistical analysis showed that treatment with either 16% or 30% carbamide peroxide resulted in a significant release of Hg from all tested amalgams (p < 0.05). There was no significant difference between 16% and 30% carbamide peroxide treatments (p > 0.05). The use of carbamide peroxide gels did not make any difference in Ag levels (p > 0.05). The release of Sn after treatment with 16% or 30% carbamide peroxide was increased (p < 0.05). The release of Cu release was decreased with tgloy and Septaloy NG 50, but increased with Dispersalloy. Additionally, Permite C (p < 0.05) SEM images revealed slight differences after treatment with 30% carbamide peroxide. Changes in the surface levels of ions were also in accordance with the ion release analysis. CONCLUSION: The release of metal ions and the microstructure of dental amalgams could be changed by treatment with 16% or 30% carbamide peroxide. PMID- 17910226 TI - Tensile bond strength and flexural modulus of resin cements--influence on the fracture resistance of teeth restored with ceramic inlays. AB - PURPOSE: This in vitro study tested tensile bond strength to enamel and dentin and the flexural modulus of three resin cements. It also determined the influence of these properties on the fracture resistance of teeth restored with ceramic inlays. MATERIALS AND METHODS: Initially, 10 standard ceramic discs were bonded to enamel using the following resin cements: Enforce (E), RelyX ARC (RX) and Fill Magic Dual Cement (FM). After seven days of storage, the specimens were subjected to tensile forces at a crosshead speed of 0.5 mm/minute until fracture. The enamel was then ground and the ceramic disks were bonded to dentin. The flexural modulus of each type of resin cement was calculated based on the straight-line tension-deformation curve using the three-point flexure method. For resistance to fracture, 40 sound maxillary premolars were randomly divided into four groups (n = 10). Three groups were submitted to preparations and restored with ceramic inlays bonded with the same resin cements used during the tensile test (n = 10). Intact teeth were used as the control group. The specimens were subjected to compressive axial loading at 0.5 mm/minute using a 10-mm steel ball until fracture. RESULTS: Statistical analysis revealed that, for all cements, the bond strength to enamel was significantly higher (p < 0.05) than that obtained in dentin. In both substrates, RX and FM showed higher bond strengths than that obtained for E (p < 0.05). In relation to flexural modulus, FM had the lowest and E the highest flexural modulus; whereas, RX differed from the other two (p < 0.05). The teeth with inlays that were bonded using RX, and E had a significantly higher (p < 0.05) fracture resistance than those where the inlays were bonded with FM but without recovering the resistance observed for the control group (intact teeth). CONCLUSION: The three resin cements had different mechanical properties. A higher flexural modulus usually resulted in improved resistance to fracture for the ceramically restored teeth. PMID- 17910227 TI - Sealing ability of dentin adhesives/desensitizer. AB - OBJECTIVES: This in vitro study evaluated the sealing ability of dentin adhesives and a desensitizer. METHODS: Standardized Class V cavities were prepared on the buccal and lingual surfaces of 55 freshly extracted human molars. The teeth were cut into two sections in a mesio-distal direction. All specimens were randomly divided into 11 groups (n = 10). In each group, the cavities were treated with one of eight different dentin adhesives, a dentin desensitizer or a special combination of these, except for the control group. After all the cavities had been stained for 24 hours with 0.5% methlyene blue solution, the teeth were longitudinally cut into two sections through the center of the cavities. Dye penetration was recorded according to the stained areas. Epoxy resin replicas of two specimens per group were analyzed by SEM. RESULTS: All dentin adhesives/desensitizer significantly reduced dentinal permeability (p < 0.05). The sealing ability of the different dentin adhesives/desensitizer was significantly different (p < 0.001). The degree of dye penetration corresponded well with the surface morphology of the dentin surfaces after the various treatments. CONCLUSIONS: None of the dentin adhesives/desensitizer could completely block fluid percolation through the dentinal tubules, but current dentin adhesives/desensitizers can significantly reduce dentin permeability. PMID- 17910228 TI - In vitro comparison of four different dental X-ray films and direct digital radiography for proximal caries detection. AB - This study investigated the efficiency of different speeds of conventional intraoral films and a direct digital system for proximal caries detection. In this study, 48 extracted human posterior permanent teeth were used. Conventional bitewing radiographs and direct digital radiographs were obtained from the teeth. Three observers independently assessed 96 proximal surfaces, each observer had 10 years of experience. The presence or absence of caries was scored according to a five-point scale. True caries depth was determined by histological examination. The diagnostic accuracy of each radiographic system was assessed by means of a receiver operating characteristic (ROC) curve analysis. The mean of areas under the ROC curve (Az) was analyzed by pairwise comparison of ROC curve. The interobserver agreement was evaluated by using ANOVA analysis. The statistical analysis of Az scores exhibited no significant difference for the five imaging modalities (p > 0.05). There was no statistically significant difference between interobserver agreements (p > 0.05). The results of this study showed that the diagnostic performance of E- and F-speed films and direct digital radiography are similar for proximal caries detection. PMID- 17910229 TI - In vitro fracture resistance of endodontically-treated maxillary premolars. AB - Many endodontically-treated teeth require quick, simple, low-cost restorations. This study evaluated the effect of horizontal pins and flowable composites on the fracture resistance of endodontically-treated maxillary premolars directly restored with resin composite. In this in vitro study, 64 intact human maxillary premolars, extracted for orthodontic reasons, were randomly divided into four groups of 16. Standard access cavities were prepared in such a way that the buccal cusp had a buccolingual thickness of 3 mm measured at the height of contour. The palatal cusp was reduced to 1.5 mm coronal to CEJ. The specimens were prepared as follows: Group 1: resin composite restoration without horizontal self-threading pins or flowable composite (control group). Group 2: resin composite restoration without horizontal self-threading pins but with a 2 mm thickness of the flowable composite. Group 3: resin composite restoration with two horizontal self-threading pins in the buccal cusp but without flowable composite. Group 4: resin composite restoration with two horizontal self threading pins in the buccal cusp and flowable composite with a thickness of 2 mm. Subsequent to thermocycling, all specimens were loaded to failure. The data were analyzed using a two-factor ANOVA test (alpha = 0.05). The maximum mean of fracture resistance was in Group 1 (632.86 +/- 119.46 N), and the minimum value was related to Group 3 (533.49 +/- 168.07 N). There was not a statistically significant difference between the groups (p > 0.05). CONCLUSION: Neither horizontal pin placement nor flowable composite had a significant effect on increasing the fracture resistance of endodontically-treated maxillary premolars restored with composite. PMID- 17910230 TI - Marginal and internal adaptation of bulk-filled Class I and Cuspal coverage direct resin composite restorations. AB - This in vitro study compared the marginal and internal adaptation of bulk-filled Class I and cuspal coverage direct resin composite restorations filled with different types of adhesive restorative systems and different thicknesses of bonding agent. Seventy-two intact, caries-free, freshly extracted human molars were randomly divided into 12 groups of six teeth each, according to the type of cavity (Class I [I] or Cuspal Coverage [C]), adhesive restorative system (SE Bond/Clearfil AP-X [SE] or Prime&Bond NT/Spectrum TPH [PB]) and thickness of bonding agent (normal or thick layer) in Class I restorations. Standardized Class I and Cuspal coverage cavities with enamel outer margins were prepared and restored with the corresponding type and thickness of bonding agent and respective resin composite. The resin composite was placed and polymerized in one increment (bulk filling). Dentinal fluid was simulated using 1:3 diluted horse serum and fed into the pulp chamber both during restoration and stressing. In six of the 12 groups, the restorations were subjected to 1.2 million mechanical occlusal cycles (maximum force 49 N; frequency 1.7Hz) and 3,000 simultaneous thermal cycles (5-50-5 degrees C). Marginal adaptation before and after mechanical and thermal stressing was assessed by using the replica technique and quantitative evaluation under SEM at 200x magnification. The teeth were dissected in a mesio-distal direction with a slow rotating diamond disc under water cooling, and the internal adaptation was also assessed by using the replica technique under the conditions described. Statistical evaluation of the continuous margin at the external and internal interface was performed with one way analysis of variance (ANOVA) and Tukey's Studentized Range (HSD) test (p = 0.05). Even though Cuspal coverage restorations (SE- C: 96.89 +/- 1.83 and PB- C: 97.15 +/- 2.93) exhibited statistically significantly better external adaptation than Class I restorations (SE- I: 63.95 +/- 12.82 and PB- I 64.74 +/- 14.62) before stressing, there was no statistically significant difference after mechanical and thermal stressing (SE- C: 76.35 +/- 18.53 and PB- C: 76.02 +/- 12.49 SE- I: 54.67 +/- 10.82 and PB- I: 59.94 +/- 15.20). After stressing, SE Bond Cuspal coverage restorations (SE- C: 96,72 +/- 3,26) exhibited superior internal adaptation compared to SE Bond Class I restorations (SE- I: 57.83 +/- 12.91). No difference was observed in internal adaptation between Prime&Bond NT Cuspal coverage and Class I restorations (PB- C:36.46 +/- 21.82, PB- I: 38.71 +/- 6.76). In Class I restorations, the increased thickness in bonding did not improve the marginal and internal adaptation either before or after stressing. Bulk-filled direct resin composite Cuspal coverage restorations exhibited marginal adaptation similar to bulk-filled direct resin composite Class I restorations. The internal adaptation of Cuspal coverage SE Bond/Clearfil AP-X restorations was superior to all the other groups tested. PMID- 17910231 TI - Amalgam and composite posterior restorations: curriculum versus practice in operative dentistry at a US dental school. AB - This study recorded the number of preclinical lecture and simulation laboratory sessions spent teaching the preparation and placement of amalgam and resin composite posterior restorations. These data were compared to the use of both materials in the operative clinic as placed by third- and fourth-year students. The number of posterior restorations inserted by the students, expressed as a function of the number of restoration surfaces, was also evaluated. The results show that the teaching of posterior restorations pre-clinically has consistently favored amalgam 2.5 to 1 during the last three years. However, clinically, resin composite is being used for posterior restorations 2.3 times more often than amalgam. The only instance that favored amalgam over composite during the last year was in the placement of four surface posterior restorations. This shift in emphasis from amalgam to composite needs to be addressed within dental educational institutions so that newly graduated dentists are prepared to place composite restorations properly. PMID- 17910232 TI - Emergency preparedness for cancer care: what have we learned? PMID- 17910233 TI - Mentors and role models influence the past, present, and future for an oncology nurse. PMID- 17910234 TI - Phase 0 trials speed early drug development. PMID- 17910235 TI - HIPAA impacts the recruitment of patients and their families to behavioral research. PMID- 17910236 TI - Help prevent an avoidable national disaster. PMID- 17910237 TI - Double trouble. Is it time to stop doing multi-patient transports? PMID- 17910238 TI - Why won't he wake up? Altered LOC, decreased respirations & pinpoint pupils provide clues to a medication mishap. PMID- 17910239 TI - Sign on the dotted line. Rules change for patient signature requirements. PMID- 17910240 TI - Tell me a story. The importance of good documentation. PMID- 17910241 TI - In the spotlight. How to develop an effective public presentation. PMID- 17910242 TI - Surviving a lawsuit. PMID- 17910243 TI - Palliative care & EMS. PMID- 17910244 TI - A sickening situation. Prehospital assessment and treatment of foodborne illnesses. AB - While nausea, vomiting and diarrhea are the most common symptoms of foodborne illness, other more serious and life-threatening conditions can occur. Foodborne illness agents can be bacterial, viral, parasitic or noninfectious. Take a thorough history when treating suspected foodborne illness to prevent further spread. Report foodborne illness through surveillance protocols in order for authorities to determine if multiple cases constitute an outbreak. PMID- 17910245 TI - Rethinking stroke care. Research shows that the treatment of hypertension in suspected stroke patients may be dangerous. PMID- 17910246 TI - Great preparations. Doing the right things before an MCI can make a big difference. PMID- 17910247 TI - Beyond the basics: COPD. PMID- 17910248 TI - Biologically mediated phosphorus precipitation in wastewater treatment with microalgae. AB - A lab-scale continuous microalgal culture was grown on sterile-filtered wastewater in order to clarify the phosphorus removing mechanisms in a microalgal treatment step that treats residual phosphorus from a hydroponic wastewater treatment pilot plant. The phosphorus assimilation was dependent on algal biomass production, whereas the chemical precipitation was dependent on phosphorus load, i.e. an increase in average precipitation rate with decreased hydraulic retention time was observed. The chemical precipitation was mainly a result of the increased pH, which was biologically mediated by the photosynthesising algae. The precipitate was composed of a calcium phosphate with magnesium included, magnesium hydroxide and calcite. A significant nitrogen removal was also experienced, which implies that the microalgal wastewater treatment is appropriate both for phosphorus and nitrogen removal. PMID- 17910249 TI - Ozone-based advanced oxidation processes in nuclear laundry water treatment. AB - The efficiency of ozone treatment in the degradation of organic compounds of nuclear laundry water is presented. Ozonation experiments were performed in different combinations of pH, hydrogen peroxide and ultraviolet radiation. The degradation of non-ionic surfactant and other organic compounds was analysed by chemical oxygen demand, total organic carbon, biochemical oxygen demand and the molecular weight distribution measurements. The optimal degradation circumstances were found to be at pH 7 with ozone, UV radiation and hydrogen peroxide addition. The transfer of ozone increased substantially thus resulting in decreased treatment time compared to our previous experiments. The reductions of chemical oxygen demand, total organic carbon and biochemical oxygen demand were about 46%, 32% and 70%, respectively. PMID- 17910250 TI - [Impact of polymetallic mine (Zn, Pb, Cu) residues on surface water, sediments and soils at the vicinity (Marrakech, Morocco)]. AB - Metal sulphide tailings present a potential risk for the environment because of their natural oxidability which leads to the production of acid mine drainage. The prospected site close to Marrakech includes zinc, lead and copper sulphide deposits. This site is located in an agricultural area near the Tensift River which is used for irrigation. In addition to the tailing leachates, underground mine waters are also discharged into the river. This represents a potential risk for the environment and human health. The aim of this study was to assess the tailings impact on surface water, sediments and soil qualities. Chemical analysis of surface water and sediments collected downstream of the mine revealed that, water and sediments present high concentrations of major ions and heavy metals. The analysis also revealed spatial as well as temporal changes in the chemical properties of the studied water and sediments. These changes are attributed to the rising phenomena. The soil near the mine presents high content of sulphate. Its Zn, Pb, Cu and Fe contents are respectively 38, 15, 11 and 1.6 times higher than non contaminated soils located far away from the site. The soil irrigated with underground mine waters shows concentrations of SO4(2-), Cu, Zn, Fe, Cd and Pb which are respectively 4, 10, 28, 2, 9 and 12 times higher than soils which are not irrigated with this mine water. This study also showed that there has been a change in the physicochemical characteristics of water and sediments in the sampling points downstream of the mine before its closure and after its activity renewal. PMID- 17910251 TI - The removal of H2S from process air by diffusion into activated sludge. AB - Emissions of H2S from publicly owned treatment works is a serious problem, therefore collection and treatment of these emissions is essential. In this work, the performance of a bench scale activated sludge system used for the removal of H2S from foul air was investigated, and the effects of H2S concentration (5 to 50 ppm,) on COD reduction and biomass settleability were studied. After biomass acclimation, the reactor was operated in a continuous mode at a hydraulic retention time of 5 h and a mean cell residence time of 6 days. Results showed that COD and H2S removal were 93.5 and 94.5%, respectively. Furthermore, H2S concentration up to 50 ppm, did not significantly affect the COD reduction. H2S loading rates of up to 7.5 mg(H2S) g(-1)MLSS, d(-1) were treated with greater than 94% efficiency. The only adverse effect of H2S that was observed was an increase in the sludge volume index at loading rates over 4.5 mg(H2S) g(-1)MLSS d(-1), at which bulking of the sludge occurred. Overall, the results indicate that H2S at concentrations usually emitted from wastewater treatment processes (lower than 50 ppm(v)), can be efficiently treated by diffusion into activated sludge without compromising the performance of the activated sludge process. PMID- 17910252 TI - Combining ultrafiltration process with coagulation pretreatment for pulp mill wastewater treatment. AB - Ultrafiltration combined with coagulation pretreatment was used to treat two kraft pulp mill wastewaters from first-stage caustic extraction and alkaline bleaching operations, respectively. Both alum and ferric chloride were tested using standard jar apparatus at different dose, pH and ionic strength conditions. Ultrafiltration tests were conducted using a crossflow flat-sheet membrane apparatus operated in the constant transmembrane pressure mode to examine the effects of membrane material, crossflow velocity and transmembrane pressure in terms of permeate flux and treated effluent quality. The results showed that coagulation with both alum and ferric chloride greatly reduced the permeate flux decline rates. In comparison with alum, greater permeate fluxes were obtained with the use of ferric chloride. Among the process parameters examined, coagulant dose was identified as the most important factor affecting the permeate flux. In addition, colour and COD removals were achieved largely by coagulation for alkaline bleaching wastewater while by membrane filtration for caustic extraction wastewater, highlighting the different mechanisms underlying contaminant removal. PMID- 17910253 TI - Fenton reaction in the presence of humates. Treatment of highly contaminated wastewater at neutral pH. AB - This paper presents the results of a treatability study conducted on highly contaminated wastewater using Fenton reaction at neutral pH, in the presence of humic substances. It has been found that addition of hurmates significantly accelerates the degradation of cyanide, thiocyanide, phenols and arsenic by ferrous iron and hydrogen peroxide. Only negligible amounts of contaminants of concern were removed when humates were added without the Fenton reagent. The mechanism of the humates effect is discussed. It is suggested that the combination of Fenton reagent with humates could provide a convenient and economical way for treatment of highly contaminated water at neutral pH. PMID- 17910254 TI - A solubility and thermodynamic study of struvite. AB - Accumulation of magnesium ammonium phosphate hexahydrate (struvite) on surfaces in contact with wastewater, especially in anaerobic sludge digestion and post digestion processes, is a widely reported problem in the wastewater treatment industry. The solubility and thermodynamic properties of struvite at different temperatures was studied. Struvite thermodynamic solubility products at temperatures between 10 and 60 'C were determined by variation of solution ionic strength and extrapolation to zero ionic strength, using an appropriate activity coefficient model. The pKsp value of struvite at 25 degrees C was found to be 13.36 (+/-0.07). The pKsp value for a temperature range of 10-60 degrees C varies from 14.36 (+/-0.05) to 14.01 (+/-0.03) with the minimum value of 13.17 (+/-0.05) at 30 degrees C. The effect of ionic strength, pH and temperature on struvite solubility was also studied. The solubility of struvite determined in deionized water was found to be 169.2 (+/-4.3) mg l(-1) at 25 degrees C, with the maximum value of 212.7 (+/-3.8) mg l(-1) at 35 degrees C. Standard enthalpy of reaction, delta Hr degrees calculated from the average Ksp values for the temperature range of 10-30 degrees C, was 23.62 Kcal mol(-1). An analytical expression for the Ksp as a function of temperature has been developed by fitting experimental data. PMID- 17910256 TI - Qualitative and quantitative determination of a humic model compound in microbial cultures by cyclic voltammetry. AB - The humic model compound, anthraquinone-2,6-disulfonate (AQDS), was characterized and measured in microbial cultures by cyclic voltammetry (CV). Under the experimental conditions, the formal reduction potential (E(o')) of the couple AQDS/AHQDS was found to be of -0.520 V vs. SCE (standard calomel electrode) at pH value of 7.0. Control experiments showed that there were no interferences of the culture medium or the microbial consortium on the quantitative determination of the quinone. The linear equation E(o') = -0.294 - 0.032 pH was found, showing that the pH used (7.0-7.8) did not affect significantly the AQDS determination by CV and AHQDS was the predominant hydroquinone formed. A calibration curve was obtained by plotting current response versus AQDS concentration with a linear correlation (r = 0.999) from 0.2 to 10 mM of AQDS. This technique was applied in a denitrifying culture to establish kinetic profiles for AHQDS formation coupled to acetate and p-cresol oxidation. CV results showed that organic matter oxidation by the denitrifying sludge was stoichiometrically associated to AQDS reduction into AHQDS-. CV was shown to be a useful tool for monitoring oxidation/reduction processes of quinones occurring in complex microbial media. PMID- 17910255 TI - Formaldehyde biodegradation and its effect on the denitrification process. AB - Simultaneous formaldehyde biodegradation and denitrification in batch assays and in a continuous lab-scale reactor were studied. In batch assays, initial biodegradation rates between 0.7 and 3.3 g CKH2O g VSS(-1) d(-1) were obtained at formaldehyde concentrations between 300 and 2150 mg l(-1). The denitrification process was affected by the presence of formaldehyde. The nitrite accumulation increased with the initial formaldehyde concentration. In the continuous reactor, removal efficiencies above 98.5% were obtained at formaldehyde loading rates between 0.37 and 2.96 kg COD m(-3) d(-1) (625-5000 mg CH2O l(-1)). Formaldehyde removal led to the appearance of methanol and formic acid in the medium. Denitrification process was almost complete (around 99.7%) at nitrogen loading rates up to 0.44 kg N-NO3- m(-3) d(-1). Nitrite occasionally appeared in the effluent at concentrations less than 2.9 mg l(-1). The composition of the biogas indicated that denitrification and methanogenesis occurred simultaneously in the same unit. PMID- 17910257 TI - Pyrolysis of waste electrical and electronic equipment: effect of antinomy trioxide on the pyrolysis of styrenic polymers. AB - This work has investigated the effect that antimony trioxide has on the pyrolysis of styrenic polymers and the effect that different types of brominated flame retardants used in plastics have on the composition of the pyrolysis products. Brominated high impact polystyrene (Br-HIPS) which contained either 5% or 0% antimony trioxide and either decabromodiphenyl oxide (DDO) or decabromodiphenyl ethane (DDE) was pyrolysed in a fixed bed reactor at 430 degrees C. Some experiments on the fixed bed reactor involved mixing the Br-HIPS with polystyrene. The gaseous products were analysed by GC-FID and GC-TCD and it was found that antimony trioxide caused an increase in the proportion of ethane and ethene and suppressed the proportion of butane and butene. When DDE was the flame retardant increased proportions of ethane and ethene were found in the pyrolysis gas compared to when DDO used. When polystyrene was mixed with the Br-HIPS it suppressed the trends observed in the gas composition during the pyrolysis of Br HIPS. The pyrolysis oils were characterised using FT-IR, GC-MS, GC-FID, and GC ECD. It was found that the plastic which did not contain antimony trioxide pyrolysed to form mainly toluene, ethylbenzene, styrene, cumene, and alpha methylstyrene. The oils produced from the pyrolysis of the plastic that contained antimony trioxide did not contain any styrene or alpha-methylstyrene, but instead contained greater concentrations of ethylbenzene and cumene. The absence of styrene and alpha-methylstyrene from the pyrolysis oil occurred even when the Br HIPS was mixed with polystyrene. GC-ECD analysis of the oils showed that the plastics which did not contain antimony trioxide pyrolysed to form (1 bromoethyl)benzene, which was totally absent from the pyrolysis oils when antimony trioxide was present in the plastic. PMID- 17910258 TI - Photocatalytic reduction of hexavalent chromium and degradation of di-n-butyl phthalate in aqueous TiO2 suspensions under ultraviolet light irradiation. AB - Simultaneous photocatalytic reduction of chromium(VI) to the less toxic Cr(III) and degradation of di-n-butyl phthalate (DBP) in aqueous TiO2, suspension under ultraviolet light irradiation was investigated. The batch experiments were carried out in a photoreactor equipped with 350 nm lamps. The system containing Cr(VI) and DBP showed the synergistic photocatalytic decontamination as DBP facilitated Cr(VI) reduction, whereas Cr(VI) accelerated DBP degradation. This study demonstrated for the first time that two important environmental pollutants Cr(VI) and DBP could be eliminated simultaneously using UV/TiO2 process. The intermediate products of DBP degradation were identified by gas chromatography mass spectrometry, and the reaction mechanism was proposed. PMID- 17910259 TI - Characteristics and anaerobic treatability of municipal and industrial estate wastewaters. AB - Centralised treatment of domestic sewage coming from A Coruna city (250000 inhabitants) and several medium and small size villages (up to 15000 inhabitants) together with industrial estate effluents was planned. The area population is about 380000 inhabitants and the overall effluent will reach about 600000 equivalent inhabitants. The characteristics of different streams will determine the treatment alternatives and pre-treatment requirements for individual streams. Domestic sewage from A Coruna showed medium to high organic content (741 mg l(-1) of COD), while domestic sewage from small villages (Lorbe, Oleiros) and industrial estates (Sabon, A Grela) showed a high variability in organic content, with COD concentration ranging from 100 to 4000 mg L(-1) or more. About 50-65% COD corresponded to suspended solids, although some industrial effluents showed lower values. Characterized industrial effluents had lower concentration of nutrients (nitrogen and phosphorus) and higher concentration of sulphate and alkalinity than domestic sewage. Anaerobic biodegradability was about 80% for domestic sewage, and ranged from 20 to 100% for industrial estate effluents. Furthermore, some industrial streams showed high microbial toxicity. The results indicate that anaerobic treatment of these effluents is of interest. However, specific pre-treatment for some individual streams or the adoption of minimisation and prevention measures in order to reach total or partial detoxification is suggested. PMID- 17910260 TI - Concurrent conditions in single cases: the need to differentiate equine dysautonomia (grass sickness) and atypical myopathy. PMID- 17910261 TI - A new method of assessing pathological elastin degradation in the horse. PMID- 17910262 TI - Do subcutaneous sutures increase risk of laparotomy wound suppuration? AB - REASONS FOR PERFORMING STUDY: Incisional drainage and suppuration occurs commonly following exploratory laparotomy; any technique with the potential to reduce the incidence of this complication warrants investigation. OBJECTIVES: To determine if abandoning the use of subcutaneous sutures in laparotomy wound closure is safe and whether it reduces the risk of suppuration. METHODS: A randomised controlled study was carried out at 2 referral hospitals in the UK, involving 309 horses undergoing exploratory laparotomy; 150 horses underwent '2-layer' closure without a subcutaneous suture, while 159 underwent conventional '3-layer' closure. Information regarding degree of oedema and gross types of discharge was recorded daily; suppuration was defined as discharge of pus. Telephone follow-up was carried out 30 days after hospital discharge to identify those complications occurring after that date and, thereafter, every 3 months. The influence of closure method on risk of wound suppuration was assessed by Chi-squared analysis and by logistic regression. Time to suppuration was modelled using a Cox proportional hazards model. RESULTS: No catastrophic failures of 2-layer closures were recorded. Prevalence of suppuration was not significantly different, being 18.7% and 23.9% for 2- and 3-layer closures, respectively (OR = 1.37, 0.79-2.37, P = 0.263). CONCLUSIONS AND POTENTIAL RELEVANCE: This study found no significant difference in prevalence or rate of wound suppuration in 2-layer closures compared to conventional 3-layer closure. Two-layer closure is recommended as a safe alternative means of achieving ventral midline abdominal closure in horses. PMID- 17910263 TI - Clinical use of the locking compression plate (LCP) in horses: a retrospective study of 31 cases (2004-2006). AB - REASONS FOR PERFORMING STUDY: Fracture repair and arthrodesis in horses require rigid fixation in order to achieve early post operative comfort. The locking compression plate (LCP) was designed to allow greater stability and less periosteal disruption than conventional plates. OBJECTIVE: To describe a series of clinical cases in which the LCP was used for fracture stabilisation or arthrodesis. METHODS: Medical records, radiographs and follow-up for 31 horses having fracture stabilisation or arthrodesis with the LCP were reviewed. Reasons for treatment included fracture of the olecranon (n = 2), metatarsal III (n = 2), ilial shaft (n = 1), femur (n = 2), radius (n = 2), medial condylar (n = 5) and complex lateral condylar (n = 1); or arthrodesis of the metacarpophalangeal joint (n = 8), proximal interphalangeal joint (n = 8) and carpus (n = 2). RESULTS: Of the 31 horses, 27 were discharged from the hospital. Complications included incisional infection (32%), implant infection (19%), implant loosening/ breakage (22%), contralateral limb laminitis (16%), colic (3%) and diarrhoea (3%). Follow up was available on all horses with at least 6 weeks of follow-up ranging up to 25 months. Of the 31 horses, 25 are sound for intended purpose, one is lame and 5 were subjected to euthanasia for complications associated with the original injury. CONCLUSIONS: The LCP is an acceptable means of fracture stabilisation and arthrodesis in the horse. POTENTIAL RELEVANCE: The LCP should be considered for difficult fracture configurations and arthrodeses, especially when increased stability of the fixation is needed. PMID- 17910264 TI - Assessment of mild hindlimb lameness during over ground locomotion using linear discriminant analysis of inertial sensor data. AB - REASONS FOR PERFORMING STUDY: Hindlimb lameness is common and can be difficult to diagnose or quantify in evaluating response to nerve blocks. An objective measure of lameness can also be used to evaluate the effectiveness of the treatment's contribution to evidence-based medicine. The inertial sensor system can be used to capture 6 degree of freedom movement during over ground locomotion and here was used to quantify tuber coxae movement in nonlame and lame horses. HYPOTHESIS: Tuber coxae movement is useful for discriminating between nonlame and lame horses. OBJECTIVES: To measure left and right tuber coxae movement in lame and nonlame horses during over ground locomotion and to implement a linear discriminant analysis to discriminate between lame and nonlame horses. METHODS: Two inertial sensors were attached to the skin over left and right tuber coxae of 21 horses (9 mildly and 12 not lame). Horses were trotted on a hard surface. A total of 1021 strides were collected. For each stride 34 features were extracted from the dorsoventral and craniocaudal movement and used in 2 different classification scenarios (lame vs. nonlame or left lame, right lame and nonlame) using linear discriminant analysis. RESULTS: Six degree of freedom inertial sensors were successfully used to collect kinematic data continuously from left and right tuber coxae in horses during over ground locomotion. These data were used for an automated classification of lameness. In the first scenario, a sensitivity of 89% was achieved with a specificity of 75%. In the second scenario, all horses could be correctly assigned to the correct class in a simple 3 class reclassification test. POTENTIAL RELEVANCE: A mobile system that reliably detects and quantifies hindlimb lameness in horses during unconstrained locomotion could be a valuable tool to perform an evidence-based assessment of lameness in horses in a clinical setting, e.g. before and after nerve blocks or before and after surgery. PMID- 17910266 TI - Antibodies to elastin peptides in sera of Belgian Draught horses with chronic progressive lymphoedema. AB - REASONS FOR PERFORMING STUDY: Chronic progressive lymphoedema (CPL) is a recently recognised disease of the lymphatic system characterised by lesions in the skin of the lower legs in several draught horse breeds, including the Belgian Draught hourse. Clinical signs slowly progress and result in severe disfigurement of the limbs. Ideally, supportive treatment should be started early in the disease process. However early diagnosis and monitoring progression of CPL is still a challenge. HYPOTHESIS: Elastin changes, characterised by morphological alterations as well as increased desmosine levels, in the skin of the distal limbs of horses affected with CPL are probably associated with a marked release of elastin degradation products, which elicit production of circulating anti elastin antibodies (AEAbs) in the serum. An enzyme-linked immunosorbent assay (ELISA) for detection of serum AEAbs may document elastin breakdown. METHODS: An ELISA technique was used to evaluate levels of AEAbs in sera of 97 affected Belgian Draught horses that were clinically healthy except for possible skin lesions, associated with CPL in their distal limbs. The horses were divided into 5 groups according to the severity of these skin lesions: normal horses (Group 1, n = 36), horses with mild lesions (Group 2, n = 43), horses with moderate lesions (Group 3, n = 8), horses with severe lesions (Group 4, n = 10) and, as a control, healthy Warmblood horses, unaffected by the disease (Group 5, n = 83). RESULTS: Horses with clinical signs of CPL had significantly higher AEAb levels compared to clinically normal Belgian Draught horses and to healthy Warmblood horses. These levels correlated with severity of lesions. CONCLUSIONS: CPL in draught horses is associated with an increase of serum AEAbs. POTENTIAL RELEVANCE: Evaluation of serum levels of AEAbs by ELISA might be a useful diagnostic aid for CPL. Pathological degradation of elastic fibres, resulting in deficient support of the distal lymphatics, is proposed as a contributing factor for CPL in Belgian Draught horses. PMID- 17910265 TI - Antibodies to elastin peptides in sera of Warmblood horses at different ages. AB - REASONS FOR PERFORMING STUDY: Early diagnosis and monitoring progression of chronic diseases in elastin-rich tissues, such as chronic progressive lymphoedema in draught horses and chronic obstructive pulmonary disease (COPD) is still a real challenge in the horse. Use of an enzyme-linked immunosorbent assay (ELISA) to detect anti-elastin antibody (AEAb) levels might be useful to assess the status of such diseases. Baseline levels, representing physiological breakdown of elastin in normal horses, are not available at present. HYPOTHESIS: Levels of AEAb in healthy horses are generally low and follow the same age-related pattern as found in man. Therefore, elevation of AEAb levels in serum can be used to evaluate pathological elastin breakdown in elastin-rich tissues. METHODS: Sera of 84 clinically healthy Warmblood horses were evaluated for the presence of AEAbs by means of a modified version of an ELISA technique used in man. The horses were divided in 5 age groups: A) < 4 months; B) 4-23 months; C) 2-3 years; D) 4-10 years; and E) > 11 years. RESULTS: Antibodies to elastin were found in all equine serum samples tested. Their levels were lowest in Group A, low in Groups B and E and highest in animals age 2-10 years. CONCLUSIONS: Measuring AEAbs in serum of horses by an ELISA technique proved to be possible and levels were stable during well-defined life stages. POTENTIAL RELEVANCE: Changes in AEAb levels are expected to be useful for early diagnosis and for monitoring progression of diseases that affect elastin-rich tissues, such as chronic progressive lymphoedema and COPD. PMID- 17910267 TI - Risk factors for Thoroughbred racehorse fatality in jump starts in Victoria, Australia (1989-2004). AB - REASONS FOR PERFORMING STUDY: The risk of fatality is greater in jump than in flat racing in Victoria, Australia. This is the first study to identify risk factors specific to jump starts in Victoria. OBJECTIVE: To identify risk factors for fatality of Thoroughbred racehorses in jump starts on all racecourses in Victoria, Australia between 1989 and 2004. METHODS: Fatalities comprised all horses that died during or immediately after a jump (hurdle or steeplechase) race or official jump trial and all horses that were subjected to euthanasia within 24 h of an event in which an injury was sustained. The retrospective study involved 191 case starts and 2324 control starts. Univariable and multivariable backward stepwise logistic regression was used to identify risk factors for fatality at any one start. A multiple level model was used with racecourse included as a random effect. RESULTS: In the final multivariable model, the duration of the racing career of the horse, the number of flat, hurdle and steeple starts accumulated in the 60 days prior to the case or control start, the number of flat and jump starts accumulated over the racing career, if the horse had had a start between 1 and 14 days prior to the case or control start, the type of jump race (hurdle or steeple), the calendar year of the start and the location of the racecourse were associated with fatality. CONCLUSIONS: The findings highlight the need to investigate further the differences between hurdle and steeplechase events and the adverse effect of prolonged prior flat racing careers on the risk of fatality in jump starts. POTENTIAL RELEVANCE: This is the first study to examine risk factors for fatality in jump starts in Victoria. The results should shape the development of interventions to reduce the risk in jump starts in the future. PMID- 17910268 TI - Risk factors for Thoroughbred racehorse fatality in flat starts in Victoria, Australia (1989-2004). AB - REASONS FOR PERFORMING STUDY: Research into risk factors specific for fatality in flat racing should be focused at a regional level as the risk factors may differ among countries and even regions within countries. OBJECTIVE: To identify risk factors for fatality of Thoroughbred racehorses in flat starts on all racecourses in Victoria, Australia between 1989 and 2004. METHODS: Fatalities comprised all horses that died during or immediately after a flat race or official flat trial, and all horses that were subjected to euthanasia within 24 h of an event in which an injury was sustained. The retrospective study involved 283 case starts and 3307 control starts. Univariable and multivariable backward stepwise logistic regression was used to identify risk factors for fatality at any one start. RESULTS: In the final multivariable model, horse gender, prior racing history, race length, racing year, racecourse location and track rating were associated with fatality. CONCLUSIONS: This study identified specific risk factors for fatality. The findings are consistent with the hypothesis that horses accumulating high speed exercise are predisposed to catastrophic injury. The study has also highlighted the need to investigate further the adverse effects of different track ratings on the incidence of injury and subsequent fatality. RELEVANCE: The results will facilitate the development of effective strategies to improve overall safety of horses and jockeys in flat racing in Victoria, Australia. PMID- 17910270 TI - Equine influenza vaccine containing older H3N8 strains offers protection against A/eq/South Africa/4/03 (H3N8) strain in a short-term vaccine efficacy study. AB - REASONS FOR PERFORMING STUDY: Surveillance of equine influenza viruses has suggested that strains included in currently licensed vaccines are a poor match for those predominantly circulating in the field. OBJECTIVE: To assess the ability of Duvaxyn IE-T Plus to provide cross protection against the newly evolved South Africa/4/03 (H3N8) strain of equine influenza virus. METHODS: The vaccine efficacy was evaluated by challenge infection with influenza strain A/eq/South Africa/4/03 (H3N8) 2 weeks after a primary course of 2 vaccinations with Duvaxyn IE-T Plus given at a 4-week interval. The outcome of challenge in vaccinated ponies was compared with that in unvaccinated animals. RESULTS: At the time of challenge, all vaccinated ponies had high levels of antibody to Newmarket/1/93, Newmarket/2/93 and South Africa/4/03 strains measured by single radial haemolysis. After challenge infection, there were statistically significantly decreased clinical scores and virus shedding was significantly lower in the vaccinated ponies compared to unvaccinated controls. CONCLUSION: Two doses of Duvaxyn IE-T Plus provides good clinical and virological protection against challenge with a variant virus 2 weeks after the 2 doses of vaccine. POTENTIAL RELEVANCE: When variant strains of equine influenza virus first emerge, booster immunisations with currently available vaccines may limit infection provided sufficiently high antibody levels are achieved, suggesting that vaccination in the face of an outbreak may be beneficial. PMID- 17910269 TI - Reproductive efficiency of Flatrace and National Hunt Thoroughbred mares and stallions in England. AB - REASONS FOR PERFORMING STUDY: Previous surveys of reproductive efficiency in British Thoroughbreds included only mares and stallions standing on studfarms in and around Newmarket. The present study was widened to compare Flatrace (FR) (Group A) and National Hunt (NH) (Group B) mares and stallions on studfarms throughout England. OBJECTIVES: To assess the influences of mare type, status and age, and veterinary manipulations on reproductive efficiency parameters. To compare the inherent fertility of stallions, based on singleton and twin pregnancy rates and pregnancy loss rates, in Groups A and B Thoroughbred breeding stock. METHODS: Managers of 24 FR and 9 NH public studfarms were asked to complete a questionnaire for each mated oestrous cycle shown by 2321 Group A and 1052 Group B mares throughout the 2002 mating season. Parameters such as per cycle singleton and twin pregnancy rates, and pregnancy loss rates were noted, and the success of hormone treatments to induce oestrus and ovulation assessed. The number of matings per oestrus and per pregnancy were recorded, together with the incidence and effectiveness of uterine and other veterinary treatments. The inherent fertility of 84 Group A and 43 Group B stallions in the study, as measured by the singleton and twin early pregnancy rates and the pregnancy loss rates recorded in the mares they mated, was also estimated. RESULTS: Per cycle early pregnancy (Days 13-16) was 63.2% for Group A and 65.3% for Group B mares; and 10.3% and 13.1%, respectively, of those pregnancies were twins or triplets. Early, middle and late pregnancy loss rates were 7.2% vs. 8.0% (Days 15-42), 3.6% vs. 6.1% (Days 42-1st October) and 2.7% vs. 2.1% (October-foaling), respectively. Matings per oestrus and per early pregnancy were significantly higher in Group B vs. Group A mares. For stallions that mated > or = 30 mares, overall early pregnancy rates per cycle in mares mated ranged from 30-89% across the 2 groups. CONCLUSIONS: No major differences in reproductive efficiency were identified between FR and NH mares and stallions. Increasing mare age was the single biggest limiting factor to an otherwise high rate of fertility in well-managed English Thoroughbreds. POTENTIAL RELEVANCE: This study identified factors that influence reproductive efficiency in the Thoroughbred. PMID- 17910271 TI - Electromyographic activity of the palatinus and palatopharyngeus muscles in exercising horses. AB - REASONS FOR PERFORMING STUDY: Determining the respiratory related activity of the palatinus and palatopharyngeus muscles in exercising horses is relevant because dysfunction of these muscles has been implicated in the pathogenesis of dorsal displacement of the soft palate. OBJECTIVE: To determine if the palatinus and palatopharyngeus muscles have respiratory activity that increases with intensity in exercising horses. METHODS: Electromyographic activity was measured in the palatinus and palatopharyngeus muscles using bipolar fine-wire electrodes while the horses completed an incremental exercise treadmill protocol. RESULTS: Both muscles displayed synchronous expiratory activity that increased significantly (P < 0.05) with exercise intensity. Phasic expiratory activity of the palatinus increased 390 +/- 98%, whereas phasic expiratory activity of the palatopharyngeus increased by 198 +/- 30% as the treadmill speed increased from 6 to 12 m/s. CONCLUSIONS: The palatinus and palatopharyngeus muscles may be important respiratory muscles, functioning to stabilise the position of the soft palate during intense exercise. CLINICAL RELEVANCE: The predominant expiratory activity of these muscles may be associated with specific muscle function related to exercise or distinct upper airway phenomena of an obligate nasal breather, such as the horse. PMID- 17910273 TI - Concurrent atypical myopathy and equine dysautonomia in two horses. AB - This report concerns 2 horses that suffered typical clinical signs of atypical myopathy (AM) and equine grass sickness (EGS) concurrently. Clinical details and pathological lesions of the cases are described. EGS and AM are relatively rare diseases and the concurrency of the diseases in the same animals is therefore considered unlikely to be a coincidence. However, it is not suggested that the evidence shows a common aetiology but rather the existence of common predisposing causes. PMID- 17910274 TI - Epidemiology of training and racing injuries. PMID- 17910272 TI - Preliminary study of mucosal IgA in the equine small intestine: specific IgA in cases of acute grass sickness and controls. AB - REASONS FOR PERFORMING STUDY: There is much evidence to suggest that group III Clostridium botulinum (types C and D) are involved in the aetiology of equine grass sickness (EGS). Antibodies have been detected previously in the blood and high levels associated with resistance to disease. Specific mucosal antibodies in the gastrointestinal (GI) tract are likely to be important in protection, and this study was performed to ascertain if such antibodies could be detected and if their levels were related to disease state. OBJECTIVES: To develop a method for quantifying IgA antibodies to C. botulinum types C and D in the GI tract of horses and to relate antibody levels to disease status. METHODS: Samples of tissue (n = 25: 6 duodenum, 7 jejunum and 12 ileum) were taken from acute grass sickness (AGS) cases and from control horses (n = 12; 4 samples from each site) at post mortem. They were extracted with the detergent saponin in the presence of protease inhibitors and assayed for total IgA, for specific IgA against botulinum neurotoxins types C and D (BoNT/C or BoNT/D), and against surface antigens of a BoNT/C negative strain of C. botulinum type C (SA) and of Clostridium tetani (TetSA), as a control. Specific IgA was expressed as percentage total IgA. RESULTS: Compared to controls, significantly higher levels of specific IgA against BoNT/C were detected in the jejunum (P = 0.04) and ileum (P = 0.02) of AGS cases. Similarly, higher specific levels against BoNT/D were demonstrated in duodenum (P = 0.01) and jejunum (P = 0.02). Significantly higher levels of IgA against SA were demonstrated only in duodenal samples (P = 0.01). CONCLUSIONS: Levels of IgA antibody to BoNTs in control horses were at near undetectable levels, suggesting no recent exposure to toxins. In AGS cases, significantly higher levels of specific IgA were detected predominantly in jejunum and ileum. POTENTIAL RELEVANCE: If specific IgA is protective then any successful vaccine for EGS should induce a mucosal response. PMID- 17910275 TI - Rhodococcus equi infection in foals: the science of 'rattles'. AB - Infection with Rhodococcus (Corynebacterium) equi is a well-recognised condition in foals that represents a consistent and serious risk worldwide. The condition manifests itself primarily as one of pulmonary abscessation and bronchitis, hence the terminology of 'rattles' derived from its most obvious clinical sign, frequently terminal when first identified. This review addresses the clinical manifestation, bacteriology and pathogenesis of the condition together with recent developments providing knowledge of the organism in terms of virulence, epidemiology, transmission and immune responses. Enhanced understanding of R. equi virulence mechanisms and biology derived from the recently available genome sequence may facilitate the rational development of a vaccine and the improvement of farm management practices used to control R. equi on stud farms in the future. Reliance on vaccines alone, in the absence of management strategies to control the on-farm challenge is likely to be disappointing. PMID- 17910276 TI - The case of Glyn vs. McGarel Groves. PMID- 17910277 TI - Effect of pH on the antimicrobial susceptibility of planktonic and biofilm-grown clinical Pseudomonas aeruginosa isolates. AB - The pH at the site of infection is one of a number of factors that may significantly influence the in vivo activity of an antibiotic prescribed for treatment of infection and it may be of particular importance in the treatment of cystic fibrosis (CF) pulmonary infection, as acidification of the airways in CF patients has been reported. As Pseudomonas aeruginosa is the most frequent causative pathogen of CF pulmonary infection, this study determines the effect that growth at a reduced pH, as may be experienced by P. aeruginosa during infection of the CF lung, has on the susceptibility of clinical P. aeruginosa isolates, grown planktonically and as biofilms, to tobramycin and ceftazidime. Time-kill assays revealed a clear loss of tobramycin bactericidal activity when the isolates were grown under acidic conditions. MIC and MBC determinations also showed decreased tobramycin activity under acidic conditions, but this effect was not observed for all isolates tested. In contrast, growth of the isolates at a reduced pH had no adverse effect on the bacteriostatic and bactericidal activity of ceftazidime. When the isolates were grown as biofilms, the pH at which the biofilms were formed did not affect the bactericidal activity of either tobramycin or ceftazidime, with neither antibiotic capable of eradicating biofilms formed by the isolates at each pH. This was in spite of the fact that the concentrations of both antibiotics used were much higher than the concentrations required to kill the isolates growing planktonically. These results show that growth in an acidic environment may reduce the susceptibility of clinical P. aeruginosa isolates to tobramycin. PMID- 17910278 TI - Efficacy of Raphanus sativus in the treatment of paracetamol-induced hepatotoxicity in albino rats. AB - In the present study, the efficacy of a methanol extract of Raphanus sativus root (RSME) is tested in albino rats that developed hepatic damage due to administration of paracetamol (100 mg/kg body weight) for 30 days. Twenty rats were divided into three experimental groups (E1, E2, E3) and one control group (EC). Two doses of RSME (80 and 120 mg/kg body weight) were administered orally to E1 and E2, respectively, and a mixture of RSME (120 mg/kg) and paracetamol (100 mg/kg) was administered to E3 for 21 days. Group EC and another group of normal rats (EN) that served as controls were administered distilled water. At the end of the experiment rats were bled to assay thiobarbituric acid reactive substances (TBARS), serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate aspartate transaminase (SGPT), reduced glutathione (GSH) and catalase. Results indicated that RSME reduced the levels of TBARS, SGOT and SGPT, and increased the level of GSH and the catalase activity in E1 and E2 as compared to the EC group. Group E3 showed decreases in TBARS, SGOT and SGPT levels, but the results were not statistically significant compared with the EN group. There was also a marked depletion in GSH level and catalase activity in this group. RSME reduced lipid peroxidation induced by paracetamol and brought the levels of SGOT and SGPT to normal, indicating liver recovery. It also brought about repletion of GSH levels and recovery of catalase activity. Results for group E3 indicated that RSME was not able to reverse the effects of paracetamol if administration continued. PMID- 17910279 TI - Relationship between level of circulating modified LDL and the extent of coronary artery disease in type 2 diabetic patients. AB - The impact of diabetes on health is due almost entirely to a series of complications that characterise the disease. It is associated with an increased incidence of macrovascular complications including coronary artery disease (CAD). The aim of the present study is to evaluate the possible relationship between the circulating levels of the modified derivatives of low-density lipoprotein (LDL) and the development of angiopathy in type 2 diabetic patients with CAD. The status of the antioxidant defences and the role of supplementation with antioxidant combinations are also studied in these patients. The study was conducted on three groups: group I (controls); group II (type 2 diabetic patients without complications--CAD[-]); and group III (including type 2 diabetic patients with stable CAD - CAD[+]). Patients in group III received adjunct treatment of antioxidant tablets for three months. The results of the present study clearly indicated that there was excessive exposure to oxidative stress in diabetic patients. The increase in free radicals was coupled with disturbance in free radical scavengers, particularly the glutathione system. The disturbance was more prominent in CAD(+) patients. The study has shown alteration in the lipid profile in diabetic groups, where the oxidised LDL (ox-LDL) levels were significantly higher than in control subjects. Diabetics with CAD had higher levels of ox-LDL than did patients without CAD. The intima/media thickness (IMT) of the carotid artery was within clinically accepted normal values if the ox-LDL level was below 100-110 u/L. Once the ox-LDL exceeded this range, IMT increased sharply with the increase in plasma ox-LDL. It seems that the level of ox-LDL should be kept below an upper limit of the 100-110 u/L range in order to avoid the serious atherosclerotic effects of this factor. The results demonstrate that plasma levels of ox-LDL correlate with the extent of coronary artery disease in type 2 diabetic patients and suggest that elevated levels of ox-LDL, can serve as an independent and significant predictor for future cardiac events in type 2 diabetic patients with CAD. PMID- 17910281 TI - Serum cystatin C, enzymuria, tubular proteinuria and early renal insult in type 2 diabetes. AB - This study investigates the association between serum cystatin C, serum creatinine concentrations, N-acetyl-beta-D-glucosaminidase (NAG enzymuria), urine alpha1-microglobulin (alpha1-MG) and beta2-microglobulin (beta2-MG) levels in subjects with type 2 diabetes (n=40, 20M/20F, age range 25-65 years; duration of diabetes 8-10 years) and age- and gender-matched healthy controls (n= 20). Exclusion criteria were absence of gross proteinuria, hypertension, dyslipidaemia or cardiovascular disease. Fasting blood samples and mid-stream specimen of urine (MSSU) were collected and serum creatinine, cystatin C, urine creatinine, NAG enzymuria, alpha1-MG and beta2-MG were measured. Diabetic subjects were separated into two groups based on albumin:creatinine concentration ratio. Group A: <3.5 (mg/mmol creatinine), group B: 3.5-35 (mg/mmol creatinine). While serum creatinine concentrations remained within the laboratory reference range for all groups, serum cystatin C concentration (mg/L) was significantly increased in group B (1.79 +/- 0.42 [mean +/- SD] compared to both control [0.81 +/- 0.10] and group A values [0.95 +/- 0.10]; both P<0.001). NAG enzymuria (units/mmol creatinine) was increased in both diabetic groups compared to control values (group B: 122 +/- 7, group A: 70 +/- 5, controls 27 +/- 2, all P<0.001). alpha1 microglobulin (microg/mmol creatinine) concentrations, similar in both the control group and group A diabetics at 1.10 +/- 0.10 and 1.11 +/- 0.21, respectively, were significantly elevated in group B at 2.10 +/- 0.41 (both P<0.01). Similarly, elevated beta2-MG (microg/mmol creatinine) levels were also observed in group B compared to both group A and control values (3.20 +/- 0.21 vs. 1.80 +/- 0.51 and 0.91 +/- 0.11, respectively; both P<0.001). In addition, group B levels were significantly higher than group A (P<0.001). These observations suggest that serum cystatin C is a more appropriate and effective biomarker for the overall estimation of GFR than serum creatinine values. In addition, increased serum cystatin C values were also associated with early renal tubular insult in subjects with type 2 diabetes, as characterised by increased NAG enzymuria, alpha1- and beta2-microglobulin excretion. PMID- 17910280 TI - Association of body mass index and Trp64Arg polymorphism of the beta3 adrenoreceptor gene and leptin level in Tehran Lipid and Glucose Study. AB - In this study the association between beta3-adrenoceptor gene polymorphism and serum concentration of leptin with body mass index (BMI) is investigated. Using subjects in the Tehran Lipid and Glucose Study, genotyping of the Trp64Arg polymorphism of the beta3-adrenoreceptor gene was performed using a restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique was used and the association with obesity was investigated. At total of 197 men and 204 women were divided into four groups (BMI<20, 20< or =BMI<25, 25< or =BMI<30, BMI< or =30) and 97, 98, 104 and 102 subjects, respectively, were placed randomly in the four groups. Leptin level was determined by an enzyme immunoassay (EIA) method and FBS, HDL-C, triglyceride and total cholesterol levels were determined by an enzyme colorimetric method. Body mass index (BMI) was also measured. The A (Arg) allele frequency was 0.08 among the population and its presence was significantly associated with increase of leptin level (AA/TA, 30.5+/-24.8 ng/mL; TT, 22.6+/-20.9 ng/mL; P=0.014) but there was no significant association with increased BMI (AA/TA, 27+/-5.6 kg/m2; TT, 25.4+/-5.5 kg/m2; P=0.072). These data show that the presence of the Arg64 allele at the beta3-adenoceptor gene locus is related to increase in leptin level in this population, but is not related to body mass index. PMID- 17910282 TI - Increased levels of soluble P-selectin correlate with iron overload in sickle cell disease. AB - Homozygous sickle cell disease (SCD) is characterised by increased soluble P selectin (sP-selectin), suggesting increased platelet activation, and high non transferrin-bound iron (NTBI), reflecting iron overload, possibly due to blood transfusion. Hypothesising a relationship between these processes, we measured both markers in 40 SCD patients and 40 age/gender/race-matched controls, finding increased levels of each marker in the patients (both P<0.001), but more pertinently a significant NTBI/sP-selectin correlation (r=0.52, P<0.001). Both indices were increased in the blood of 15 recently-transfused patients compared with 25 three-month transfusion-free patients (P<0.001), but only sP-selectin was higher in present sickle crisis (P<0.001). We suggest that increased NTBI associated with blood transfusion iron overload in SCD may promote platelet activation. PMID- 17910283 TI - Inquilinus limosus isolated from a cystic fibrosis patient: first UK report. PMID- 17910285 TI - Enhancement of diaminobenzidine staining of chorioretinal specimens by cobaltous ions. PMID- 17910284 TI - Environmental persistence of Pseudomonas aeruginosa and Burkholderia multivorans in sea water: preliminary evidence of a viable but non-culturable state. PMID- 17910286 TI - Molecular detection and identification of Cryptosporidium species in lettuce employing nested small-subunit rRNA PCR and direct automated sequencing. PMID- 17910287 TI - Diagnosis, treatment and management of venous thromboembolism: recent developments relevant to biomedical scientists. AB - Three documents from government-sponsored bodies have recently provided new guidance on the diagnosis, treatment and management of venous thromboembolism. The Report of the Independent Expert Working Group to the Chief Medical Officer makes recommendations on general administrative arrangements, and provides a strategy for thromboprophylaxis. Among the recommendations of Guideline 46 from the National Institute for Health and Clinical Excellence are that all patients about to undergo surgery should be assessed to identify their risk factors for developing veno-thromboembolic disease, and be offered graduated compression/anti embolism stockings and/or pharmacoprophylaxis. The National Patient Safety Agency document focuses principally on the management of, and education in, the use of oral anticoagulants. The impact and implications of these three documents for haematology-based biomedical scientists, such as in leading a thrombosis team, directing clinical management, training of healthcare professions, and in patient education, will be discussed. PMID- 17910288 TI - Merits of breastfeeding children through the toddler years. PMID- 17910289 TI - Emergency preparedness plan crucial for physicians and patients. PMID- 17910290 TI - Current understanding of tendinopathies and treatment options. PMID- 17910291 TI - Principles of the patient-centered medical home. PMID- 17910292 TI - Improving care with the patient-centered medical home. PMID- 17910293 TI - Hard choices. PMID- 17910294 TI - Gout: an update. AB - Arthritis caused by gout (i.e., gouty arthritis) accounts for millions of outpatient visits annually, and the prevalence is increasing. Gout is caused by monosodium urate crystal deposition in tissues leading to arthritis, soft tissue masses (i.e., tophi), nephrolithiasis, and urate nephropathy. The biologic precursor to gout is elevated serum uric acid levels (i.e., hyperuricemia). Asymptomatic hyperuricemia is common and usually does not progress to clinical gout. Acute gout most often presents as attacks of pain, erythema, and swelling of one or a few joints in the lower extremities. The diagnosis is confirmed if monosodium urate crystals are present in synovial fluid. First-line therapy for acute gout is nonsteroidal anti-inflammatory drugs or corticosteroids, depending on comorbidities; colchicine is second-line therapy. After the first gout attack, modifiable risk factors (e.g., high-purine diet, alcohol use, obesity, diuretic therapy) should be addressed. Urate-lowering therapy for gout is initiated after multiple attacks or after the development of tophi or urate nephrolithiasis. Allopurinol is the most common therapy for chronic gout. Uricosuric agents are alternative therapies in patients with preserved renal function and no history of nephrolithiasis. During urate-lowering therapy, the dose should be titrated upward until the serum uric acid level is less than 6 mg per dL (355 micromol per L). When initiating urate-lowering therapy, concurrent prophylactic therapy with low-dose colchicine for three to six months may reduce flare-ups. PMID- 17910295 TI - Information from your family doctor. Gout: what you should know. PMID- 17910296 TI - Diagnosis and management of metatarsal fractures. AB - Patients with metatarsal fractures often present to primary care settings. Initial evaluation should focus on identifying any conditions that require emergent referral, such as neurovascular compromise and open fractures. The fracture should then be characterized and treatment initiated. Referral is generally indicated for intra-articular or displaced metatarsal fractures, as well as most fractures that involve the first metatarsal or multiple metatarsals. If the midfoot is injured, care should be taken to evaluate the Lisfranc ligament. Injuries to this ligament require referral or specific treatment based on severity. Nondisplaced fractures of the metatarsal shaft usually require only a soft dressing followed by a firm, supportive shoe and progressive weight bearing. Stress fractures of the first to fourth metatarsal shafts typically heal well with rest alone and usually do not require immobilization. Avulsion fractures of the proximal fifth metatarsal tuberosity can usually be managed with a soft dressing. Proximal fifth metatarsal fractures that are distal to the tuberosity have a poorer prognosis. Radiographs should be carefully examined to distinguish these fractures from tuberosity fractures. Treatment of fractures distal to the tuberosity should be individualized based on the characteristics of the fracture and patient preference. Nondisplaced fractures of the proximal portion of metatarsals 1 through 4 can be managed acutely with a posterior splint followed by a molded, non-weight-bearing, short leg cast. If radiography reveals a normal position seven to 10 days after injury, progressive weight bearing may be started, and the cast may be removed three to four weeks later. PMID- 17910297 TI - Ocular emergencies. AB - Prompt recognition and appropriate treatment of ocular emergencies are essential in the primary care setting when the outcome may depend on timely management. All ocular emergencies, including a penetrating globe injury, retinal detachment, central retinal artery occlusion, acute angle-closure glaucoma, and chemical burns, should be referred immediately to the emergency department or an ophthalmologist. Careful eye examination and simple tests can help primary care physicians make decisions about appropriate treatment and referral. All patients with eye problems should be tested for visual acuity and ocular movements. Confrontation visual field examination, pupillary examination, and direct ophthalmoscopy of both eyes also should be performed. Ocular injury from high velocity trauma or from chemicals may be easily misdiagnosed. After a chemical burn, thorough eye washing for at least 30 minutes or until the pH of the eye is within physiologic range is critical to prevent further damage. Use of an eye shield is required in patients with a ruptured globe to protect the injured eye and preserve the patient's vision. PMID- 17910298 TI - Treatment of lateral epicondylitis. AB - Lateral epicondylitis is a common overuse syndrome of the extensor tendons of the forearm. It is sometimes called tennis elbow, although it can occur with many activities. The condition affects men and women equally and is more common in persons 40 years or older. Despite the prevalence of lateral epicondylitis and the numerous treatment strategies available, relatively few high-quality clinical trials support many of these treatment options; watchful waiting is a reasonable option. Topical nonsteroidal anti-inflammatory drugs, corticosteroid injections, ultrasonography, and iontophoresis with nonsteroidal anti-inflammatory drugs appear to provide short-term benefits. Use of an inelastic, nonarticular, proximal forearm strap (tennis elbow brace) may improve function during daily activities. Progressive resistance exercises may confer modest intermediate-term results. Evidence is mixed on oral nonsteroidal antiinflammatory drugs, mobilization, and acupuncture. Patients with refractory symptoms may benefit from surgical intervention. Extracorporeal shock wave therapy, laser treatment, and electromagnetic field therapy do not appear to be effective. PMID- 17910299 TI - Information from your family doctor. Exercises for tennis elbow. PMID- 17910300 TI - Information from your family doctor. Tennis elbow: what you should know. PMID- 17910301 TI - Acute red eye. Pingueculitis. PMID- 17910302 TI - Central service in the regulatory spotlight. Interview by Alan Joch. PMID- 17910303 TI - Vendors' contribution to infection control. PMID- 17910304 TI - Is that product safe? PMID- 17910305 TI - 2007 Most Wired Survey. Innovators plug in systems to drive supply chain efficiency. PMID- 17910306 TI - Making an intelligent smart pump purchase. PMID- 17910308 TI - Internet negotiating pays big dividends. PMID- 17910307 TI - Extended cycles and flash sterilization. PMID- 17910309 TI - APIC study delves into MRSA in hospital settings. PMID- 17910310 TI - Role of stem cell transplant in pediatric cancers. PMID- 17910311 TI - Targeted therapy in rectal cancer. AB - Epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) are often overexpressed in colorectal cancer and are associated with inferior outcomes. Based on successful randomized phase III trials, anti-EGFR and anti-VEGF therapeutics have entered clinical practice. Cetuximab (Erbitux), an EGFR-specific antibody, is currently approved in the United States in combination with irinotecan (Camptosar) for patients with metastatic colorectal cancer refractory to irinotecan or as a single agent for patients unable to tolerate irinotecan-based therapy. In retrospective analyses, patients with EGFR expressing rectal cancer undergoing neoadjuvant radiation therapy had a significantly inferior disease-free survival and lower rates of achieving pathologic complete response. Based on the positive data in metastatic colorectal cancer and synergy with radiation therapy seen in preclinical models, there is a strong rationale to combine cetuximab with neoadjuvant radiation therapy and chemotherapy in rectal cancer. Bevacizumab (Avastin), a VEGF-specific antibody, was the first antiangiogenic agent to be approved in the United States for use in combination with standard chemotherapy in the first- and second-line of treatment in metastatic colorectal cancer. VEGF-targeted therapy may lead to indirect killing of cancer cells by damaging tumor blood vessels, and may increase the radiosensitivity of tumor-associated endothelial cells. VEGF blockade can also "normalize" tumor vasculature, thereby leading to greater tumor oxygenation and drug penetration. This review will address completed and ongoing trials that have established and continue to clarify the effects of these agents in rectal cancer. PMID- 17910312 TI - Therapeutic options in gastric cancer: neoadjuvant chemotherapy vs postoperative chemoradiotherapy. AB - The majority of patients who undergo resection for gastric cancer experience relapse and ultimately die of their disease. Therefore, considerable attention has been paid to neoadjuvant and adjuvant strategies to improve surgical outcomes. Two different approaches have been tested in major clinical trials conducted in the past several years: Postoperative chemoradiotherapy was assessed in a US Southwest Oncology Group/Intergroup study (SWOG 9008/INT 0116), and perioperative chemotherapy was studied in a UK Medical Research Council (MRC) randomized trial (the MRC Adjuvant Gastric Infusional Chemotherapy [MAGIC] trial). These trials demonstrated statistically significant survival benefits in patients with resectable gastric cancer. This review will consider these trials and their implications for clinical practice. PMID- 17910313 TI - Non-Hodgkin's lymphoma in the elderly. Part 1: Overview and treatment of follicular lymphoma. AB - Non-Hodgkin's lymphoma is one of a few malignancies that have been increasing in incidence over the past several decades. Likewise, these disorders are more common in elderly patients, with a median age of occurrence of 65 years. Therapy in elderly patients may be affected by multiple factors, especially attendant comorbidities. The approaches to management of these patients, with either indolent or aggressive disease processes, have been based on prospective clinical trial results, many of which have included a younger patient population. Fortunately, over the past decade, results of treatment trials that have targeted an older patient population have emerged. The disease incidence and treatment approaches for both follicular (part 1 of this article) and diffuse aggressive (part 2) histologies in elderly patients are reviewed, as well as the impact of aging on the care of these patients. PMID- 17910314 TI - Neoadjuvant therapy for hepatocellular carcinoma: is there an optimal approach? AB - Hepatocellular carcinoma (HCC) is an aggressive tumor that often occurs in the setting of chronic liver disease and cirrhosis. The incidence of hepatocellular carcinoma is increasing in the United States and worldwide. Orthotopic liver transplantation (OLT) is a viable and potentially curative option for selected patients with HCC. Locoregional therapy has been used to control HCC before transplantation because of the limited number of donor organs, to prevent tumor progression, and to decrease the incidence of dropouts from the transplant waiting list. Traditionally, multiple investigational locoregional modalities such as tumor resection, radiofrequency ablation, transarterial chemoembolization, and systemic chemotherapy have been used as "bridging" therapies. While the investigation of novel neoadjuvant treatments is justified in an effort to prevent tumor progression, the absence of randomized controlled trials leaves uncertainty about the utility of these maneuvers in improving outcome. This review summarizes the current data on the different modalities used worldwide in the neoadjuvant treatment of hepatocellular carcinoma, the rationale for these approaches, efficacy, potential complications, and future prospects. PMID- 17910316 TI - Transparency: it's about the customers. PMID- 17910315 TI - Problem-related distress in cancer patients drives requests for help: a prospective study. AB - The Moores UCSD Cancer Center has implemented the use of an innovative instrument for screening cancer patients at first visit to assist them with distress due to cancer-related problems. This 36-question screening instrument addresses physical, practical, social, psychological and spiritual problems. Patients are asked to rate the severity of each problem on a scale of 1 to 5, and to circle "Yes" if they would like staff assistance. Data from a prospective study of the first 2,071 patients to complete this questionnaire has been entered into a database and analyzed to identify common patient problems, demographics, and trends. The five most common causes of problem-related distress were fatigue, sleeping, finances, pain, and controlling my fear and worry about the future. The five most common problems for which patients circled "Yes" to ask for assistance were understanding my treatment options, fatigue, sleeping, pain, and finances. Compared to the entire population, patients who circled "Yes" on a particular problem, demonstrated a robust increase in problem-related distress. PMID- 17910317 TI - Creating large group relationships. PMID- 17910318 TI - Why is tracking ROI more challenging in healthcare? PMID- 17910319 TI - Strategies for recruiting and retaining top hospitalists. PMID- 17910320 TI - Seize your one chance to make an impression. PMID- 17910321 TI - Pussy Willow. Salix discolor. PMID- 17910322 TI - Practical aspects of ambulatory diagnoses and management of immunodeficiency disorders. PMID- 17910323 TI - Cross-reactivity of pollen allergens: impact on allergen immunotherapy. AB - OBJECTIVE: To provide guidelines for the rational formulation of allergen immunotherapy extracts based on knowledge of pollen allergen and epitope cross reactivity. DATA SOURCES: A PubMed search was performed for articles published from 1966 to 2007 using the keywords pollen, allergen, and cross-reactivity. Older literature was found through cross-referencing of older articles and older reviews on pollen cross-reactivity. STUDY SELECTION: Articles that dealt with crude pollen extracts and characterized allergens that addressed cross-reactivity were selected for inclusion in this review. RESULTS: In addition to unique allergens, several families of botanic proteins have similarities that allow them to act as pan-allergens. Although frequently these are minor allergens, in some circumstances they may also be major allergens. Recent studies have investigated nonspecific lipid transfer proteins, calcium-binding proteins, pathogenesis related protein families, and profilins. Calcium-binding proteins and nonspecific lipid transfer proteins are responsible for pollen-fruit interactions and pollen cross-reactivity. Clarification of pollen allergen enzymatic activity helps explain the ubiquitous nature of these proteins. CONCLUSION: Characterization of specific pollen allergens and their protein families has provided insight into cross-reactivity. Clarification of these relationships allows for consolidation or substitution in formulation of inhalant extracts. PMID- 17910324 TI - Interleukin 13 and the beta-adrenergic blockade theory of asthma revisited 40 years later. AB - BACKGROUND: Beta2-Adrenergic agonists are the most potent agents clinically used in inhibiting and preventing the immediate response to bronchoconstricting agents and in inhibiting mast cell mediator release. This raises the possibility that an abnormality in beta-adrenergic receptor function or circulating catecholamine levels could contribute to airway hyperresponsiveness. OBJECTIVE: To link interleukin 13 (IL-13) to the pathogenesis of asthma. METHODS: Almost 4 decades ago, Andor Szentivanyi published a beta-adrenergic theory of atopic abnormality in bronchial asthma. He proposed 9 characteristics to define bronchial asthma. Because he published these 9 tenets of the beta-adrenergic blockade theory of asthma in 1968, it is appropriate and important to evaluate their relevance in light of advances in pharmacology, inflammation, and immunology. RESULTS: We describe the effects of the allergic reaction on beta-adrenergic responses and airway responsiveness. Both IL-1beta and tumor necrosis factor a have been detected in increased amounts in bronchial lavage fluids in allergic airway inflammation. Both IL-13 and the proinflammatory cytokines IL-1beta and tumor necrosis factor a have been demonstrated in airway smooth muscle to cause a decreased relaxation response to beta-adrenergic agonist. However, IL-13 has been shown to be necessary and sufficient to produce the characteristics of asthma. CONCLUSION: The decreased adrenergic bronchodilator activity and associated hypersensitivity to mediators put forth by Szentivanyi can be elicited with IL-13 and support its role in the pathogenesis of asthma. PMID- 17910325 TI - Short-term effect of pollen exposure on antiallergic drug consumption. AB - BACKGROUND: Several studies have investigated the association between pollen exposure and asthma emergency admissions, but only 2 have investigated the effect of airborne allergens on consultations for rhinitis or conjunctivitis and none has used drug consumption as the health indicator. OBJECTIVE: To analyze the short-term association between pollen exposure and antiallergic drug consumption in the urban area of Clermont-Ferrand, France, taking into account the potentially confounding effect of air pollution and meteorological factors. METHODS: We used the French health insurance database to select all individuals from the Clermont-Ferrand urban area having benefited from reimbursement for antiallergic treatment from January 1, 2000, through December 31, 2001, and from January 1, 2003, through December 31, 2004. An episode of treated allergic rhinitis, rhinosinusitus, or conjunctivitis (ARC) was defined as the association of an oral antihistamine and a local antiallergic drug on the same prescription. The relations between daily changes in pollen concentrations and daily changes in the number of treated ARC cases were analyzed using a Poisson regression model with penalized spline functions. RESULTS: The risk of treated ARC associated with an interquartile increase in pollen concentration increased significantly for Poaceae (5%, P < .001), Fraxinus (7%, P < .001), Betula (7%, P < .001), and Corylus (2%, P < .02). This increase was significant in all age groups for Poaceae and Fraxinus pollen and in people younger than 65 years for Betula pollen. The effect was mainly concentrated on the present day, except for Poaceae pollens, for which the risk remained significantly (P < .001) increased until 3 days lag time. CONCLUSIONS: This study showed a significant increase in treated ARC cases related to Poaceae, Fraxinus, and Betula. Specific risks are difficult to evaluate for species that share the same pollination period. Time-series studies based on drug consumption are useful to highlight and to supervise pollen related diseases requiring ambulatory care. PMID- 17910326 TI - Validity and reproducibility of morphologic analysis of nasal secretions obtained using ultrasonic nebulization of hypertonic solution. AB - BACKGROUND: Collection of nasal secretions is important for the evaluation of upper airways inflammation in many nasal disorders. OBJECTIVE: To study the validity and reproducibility of nasal secretion cellularity induced by nebulization of hypertonic solution in patients with allergic rhinitis (AR), patients with nonallergic rhinitis with eosinophilia syndrome (NARES), and control subjects. METHODS: Sixty-eight individuals (29 with AR [mean +/- SD age, 33.3 +/- 16.9 years], 23 with NARES [mean +/- SD age, 46.4 +/- 16.6 years], and 16 controls [mean +/- SD age, 42.1 +/- 15.1 years]) underwent ultrasonic nebulization of hypertonic (4.5%) saline solution on 2 different occasions to study the validity and reproducibility of total and differential cell counts of nasal secretions. RESULTS: The mean +/- SD percentage of eosinophils was significantly higher in samples from patients with AR (20.8% +/- 23.1%) and NARES (18.7% +/- 22.8%) than in samples from controls (0.6% +/- 0.6%; P < .001 for both). There was a significant correlation between 2 samples of nasal secretions obtained on 2 different occasions for percentages of macrophages, neutrophils, eosinophils, and epithelial cells. CONCLUSIONS: The analysis of nasal secretions obtained using ultrasonic nebulization of hypertonic solution can distinguish patients with AR and NARES from controls. The reproducibility of this technique is good for macrophages, neutrophils, eosinophils, and epithelial cells. This method could be used to detect nasal airway inflammation in clinical settings. PMID- 17910327 TI - Associations of physician-diagnosed asthma with country of residence in the first year of life and other immigration-related factors: Chicago asthma school study. AB - BACKGROUND: Among Mexican Americans in the United States, US-born children have higher rates of asthma than their Mexico-born peers. OBJECTIVE: To evaluate the associations of immigration-related variables with physician-diagnosed asthma in a sample of Mexican American children. METHODS: We analyzed data from the ongoing Chicago Asthma School Study, a population-based cross-sectional study, for 10,106 Mexican American schoolchildren in Chicago, Illinois. RESULTS: Mexican American children who lived in the United States in the first year of life were more likely to have physician-diagnosed asthma than their peers who lived in Mexico in the first year of life, independent of age, sex, income, language, and country of birth (odds ratio [OR], 1.79; 95% confidence interval [CI], 1.09-2.94). The risk of asthma in US-born children was higher (but not significantly) than that observed in Mexico-born children after accounting for covariates, including country of residence in the first year of life (OR, 1.37; 95% CI, 0.86-2.18). Long-term immigrants (lived in the United States for 10 years) had an increased risk of asthma compared with short-term immigrants (lived in the United States for <10 years), independent of country of residence in the first year of life (OR, 1.93; 95% CI, 1.00-3.73). CONCLUSION: These findings confirm the importance of early childhood exposures and environmental factors that are modified with migration and acculturation in asthma development. PMID- 17910328 TI - Circulating neutrophil CD14 expression and the inverse association of ambient particulate matter on lung function in asthmatic children. AB - BACKGROUND: Identifying baseline inflammatory biomarkers that predict susceptibility to size-specific particulate matter (PM) independent of gaseous pollutants could help us better identify asthmatic subpopulations at increased risk for the adverse health effects of PM. OBJECTIVE: To evaluate whether the association between lung function and exposure to ambient levels of PM less than 2.5 microm in diameter (PM2.5) (fine) and 10 to 2.5 microm in diameter (PM(10 2.5)) (coarse) in children with persistent asthma differed across baseline measures of inflammation and innate immune activation. METHODS: We performed a panel study on a local population of 16 children with persistent asthma and evaluated daily pulmonary function (percentage of predicted peak expiratory flow and forced expiratory volume in 1 second) while concurrently measuring daily PM2.5 and PM(10-2.5) exposure from a central site in Chapel Hill, North Carolina. The children underwent a baseline medical evaluation that included assessment of several immunoinflammatory biomarkers in peripheral blood. RESULTS: Children without measurable CD14 expression on circulating neutrophils had significantly reduced pulmonary function (forced expiratory volume in 1 second and peak expiratory flow) with each interquartile range (IQR) increase in PM2.5 (IQR = 8.5 microg/m3) and PM(10-2.5) (IQR = 4.1 microg/m3) concentration, unlike children with measurable CD14 expression (P < .001 for interaction). CONCLUSIONS: Asthmatic children with muted surface expression of CD14 on circulating neutrophils may have a decreased capacity to respond to bacterial components of PM. PMID- 17910329 TI - Application of the 16-kDa buckwheat 2 S storage albumin protein for diagnosis of clinical reactivity. AB - BACKGROUND: The 16-kDa protein of buckwheat (BW) has been implicated as a major allergen in BW allergy. OBJECTIVE: To characterize the 16-kDa allergen and evaluate its clinical significance as an indicator of BW allergy. METHODS: Complementary DNA from the 16-kDa allergen was cloned and expressed in Escherichia coli. Allergenicity was confirmed with IgE immunoblotting or with an enzyme-linked immunosorbent assay. The clinical utility of the recombinant protein (r16 kDa) for diagnosis of BW reactivity was evaluated in 18 BW-allergic and in 20 asymptomatic BW-sensitized subjects. RESULTS: The 16-kDa allergen, composed of 127 amino acids, has 50% homology to the reported 8-kDa BW allergen, which belongs to the 2 S storage albumin. The r16-kDa protein can inhibit specific IgE (sIgE) antibody binding to the native BW 16-kDa allergen but minimally inhibited sIgE binding to crude BW extract. Approximately 77.8% of patients with the BW allergy produced sIgE antibodies to the r16-kDa protein, compared with a complete lack of reactivity in the 20 asymptomatic BW-sensitized subjects. The areas of the receiver operating characteristic curves for the skin prick test (mean, 0.93; 95% confidence interval, 0.85 to approximately 1.01; P < .001) and the rl6-kDa enzyme-linked immunosorbent assay (mean, 0.93; 95% confidence interval, 0.84 to approximately 1.01; P < .001) were higher than the area of the BW IgE measurement curve determined by ImmunoCAP (a system for assaying serum IgE) (mean, 0.80; 95% confidence interval, 0.66 to approximately 0.94; P = .002). CONCLUSIONS: The 16-kDa allergen belongs to the 2 S storage albumin. Measurement of rl6-kDa sIgE was more discriminating than measurement of ImmunoCAP sIgE in whole BW extracts for the diagnosis of clinical reactivity to BW. PMID- 17910330 TI - H1-antihistamine treatment in young atopic children: effect on urticaria. AB - BACKGROUND: There are few published, randomized, double-masked, placebo controlled, clinical trials of interventions for urticaria in the pediatric population. OBJECTIVE: To study the effect of long-term treatment with the H1 antihistamine levocetirizine on urticaria in young atopic children. METHODS: In the randomized, double-masked, parallel-group Early Prevention of Asthma in Atopic Children Study, children with atopic dermatitis aged 12 to 24 months at enrollment received levocetirizine, 0.125 mg/kg, or matching placebo twice daily for 18 months. On a diary card, the child's caregiver recorded the days on which urticaria was observed. This was validated by the study investigator and entered into the electronic case report form, along with any additional relevant information. RESULTS: A total of 510 atopic children (mean +/- SEM age, 19.4 +/- 0.2 months) composed the intention-to-treat population. During the subsequent 18 months, 27.5% (70/255) of the children taking levocetirizine and 41.6% (106/255) of the children taking placebo experienced urticaria (P < .001). The mean +/- SEM number of urticaria episodes was 0.71 +/- 0.11 in those receiving levocetirizine and 1.71 +/- 0.25 in those receiving placebo (P < .001). The mean +/- SEM duration of urticaria episodes was 4.43 +/- 1.57 days in those receiving levocetirizine and 5.36 +/- 1.27 days in those receiving placebo (P < .001). CONCLUSIONS: Urticaria is common in atopic toddlers and deserves recognition as an important disorder that occurs early in the atopic marathon. Regular long-term treatment with levocetirizine effectively prevents and treats urticaria in young children. The results of this study strengthen the evidence base for the use of relatively nonsedating, second-generation H1-antihistamines in the pediatric population. PMID- 17910331 TI - Efficacy and safety of azelastine nasal spray at a dose of 1 spray per nostril twice daily. AB - BACKGROUND: Azelastine hydrochloride nasal spray is available worldwide for the treatment of seasonal allergic rhinitis (SAR) and perennial allergic rhinitis. One spray per nostril twice daily is the most commonly recommended dose. OBJECTIVE: To determine the efficacy and safety of azelastine nasal spray, 1 spray per nostril twice daily, in patients with SAR. METHODS: In 2 studies conducted in the United States we assessed 554 patients with moderate-to-severe SAR who were still symptomatic after a 1-week placebo lead-in period. Patients were randomized to 2 weeks of double-blind treatment with azelastine nasal spray, 1 spray per nostril twice daily, or placebo nasal spray. The primary efficacy variable was change from baseline in total nasal symptom score, consisting of sneezing, itchy nose, runny nose, and nasal congestion. RESULTS: Mean differences in total nasal symptom score between the azelastine and placebo groups were significant in both studies: 2.69 vs 1.31 (P = .01) in study 1 and 3.68 vs 2.50 (P = .02) in study 2. Bitter taste was reported by 8.3% of patients treated with 1 spray per nostril twice daily compared with the labeled incidence of 19.7% with 2 sprays per nostril twice daily. Somnolence was reported by 1 patient (0.4%) using the 1-spray regimen compared with the labeled incidence of 11.5% using the 2-spray regimen. CONCLUSIONS: Azelastine nasal spray at a dose of 1 spray per nostril twice daily is effective and has improved tolerability compared with 2 sprays per nostril twice daily in patients with SAR. PMID- 17910332 TI - Immunotherapy with mosquito (Culex quinquefasciatus) extract: a double-blind, placebo-controlled study. AB - BACKGROUND: Mosquito allergy is well established, but mosquito immunotherapy requires validation using clinical and immunologic variables. OBJECTIVE: To evaluate the tolerability and efficacy of specific immunotherapy with Culex quinquefasciatus (mosquito) extract. METHODS: We performed a randomized, double blind, placebo-controlled trial of immunotherapy for 1 year in 40 patients with asthma, rhinitis, or both. Patients were evaluated by means of intradermal testing, symptom and drug scores, and histamine provocation testing before and after 1 year of immunotherapy. Mosquito specific IgE and IgG subclass antibody responses were evaluated at the basal level and after 1 year. RESULTS: Patients receiving allergen immunotherapy for 1 year showed a significant improvement compared with baseline and patients receiving placebo regarding skin reactions, symptom scores (rhinitis and asthma), and forced expiratory volume in 1 second. Provocation concentration of histamine that caused a decrease in forced expiratory volume in 1 second of 20% by inhalation was elevated in the group receiving immunotherapy. In the active group serologic analysis showed a slight reduction in IgE levels (P = .02) but a significant elevation in IgG4 levels (P = .001), with a significant decrease in the IgE/IgG4 ratio (P = .001). All these changes in the placebo group were nonsignificant. CONCLUSIONS: Allergen immunotherapy with mosquito extract was well tolerated, with improvement in symptoms and airway reactivity. Good clinical outcome was associated with increased IgG4 antibody levels. PMID- 17910333 TI - Assessment and clinical interpretation of reduced IgG values. PMID- 17910334 TI - Anaphylaxis to Hippobosca equina (louse fly). AB - BACKGROUND: IgE-mediated allergy to insects different from Hymenoptera species is seldom reported. OBJECTIVE: To describe and study the case of a previously nonatopic man with an anaphylactic reaction (grade III, Mueller) caused by a bite from a louse fly (Hippobosca equina). METHODS: In vivo (skin prick tests) and in vitro (enzyme-linked immunosorbent assay and sodium dodecyl sulfatepolyacrylamide gel electrophoresis immunoblotting) tests were used for diagnosis. Cross reactivity between H. equina and different insects was investigated. RESULTS: Results of skin prick tests and serum specific IgE were positive to H. equina. Immunoblot inhibition studies identified common bands in H. equina, Apis mellifera, and Musca domestica, but this cross-reactivity did not affect a band of 16 or 15 kDa. This molecular weight is similar to that of phospholipase A2 in A mellifera venom. CONCLUSION: We describe a case of IgE-mediated allergy to H. equina in which specific molecular band proteins seemed to be responsible for the reaction. PMID- 17910335 TI - Intrapartum anaphylaxis to penicillin in a woman with rheumatoid arthritis who had no prior penicillin allergy. AB - BACKGROUND: Little is known about the development of drug allergy during pregnancy or in patients with altered immune status. OBJECTIVE: To report a case of new-onset penicillin allergy during pregnancy in a woman with rheumatoid arthritis. METHODS: A 39-year-old woman with rheumatoid arthritis developed intrapartum anaphylaxis that led to fetal demise. She had previously received penicillin-based antibiotics without any allergic reactions. Because of group B streptococcus colonization, an intravenous infusion of penicillin G was started during labor. Within minutes, she developed severe anaphylaxis. RESULTS: A fluorescent enzyme immunoassay revealed a moderate level of specific IgE to penicilloyl G and penicilloyl V (3.15 kU/L and 2.77 kU/L, respectively). Given the patient's history, these positive results were considered confirmatory of penicillin allergy. This case raises a number of salient points. First, patients can develop severe allergy to penicillin despite having safely received penicillins in the past. Possible factors that influenced the development of severe penicillin sensitivity in this patient are discussed. Second, unexpected intrapartum anaphylaxis can occur, which can be life threatening to the mother or fetus. Third, safe and reliable methods for diagnosis of drug allergy must be available. CONCLUSIONS: This case illustrates that during the current unavailability of skin testing reagents in the United States, a positive result on in vitro testing can be helpful in confirming penicillin allergy in cases in which drug challenge is deemed unsafe. PMID- 17910336 TI - Risk of home immunotherapy. PMID- 17910337 TI - Fine needle aspiration cytology of basal cell adenoma of the salivary gland. AB - OBJECTIVE: To formulate cytologic features for differential diagnosis of basal cell adenoma (BCA). STUDY DESIGN: The usefulness of 5 items for a cytologically definitive diagnosis of BCA was examined. The 5 items in 8 BCA and 22 non-BCA cases (adenoid cystic carcinoma [ADCC], basal cell adenocarcinoma, myoepithelioma, pleomorphic adenoma and polymorphous low-grade adenocarcinoma) that displayed mimicking cytology were examined cytologically. RESULTS: The useful items were < 5.1 microm in mean of epithelial nuclear short diameter; mild atypia on definitive diagnosis; stromal cell cluster combining smooth margin surrounding the epithelial cell cluster or containing the epithelial cell cluster; epithelial clusters surrounded by or adhered to a thick, hyalinized smooth margin without stromal cluster; and closely fastened, tight clusters with denser cytoplasm than ADCC, but an indistinct border, with oval nuclei and no hyaline cells. CONCLUSION: Five items are useful criteria for BCA. PMID- 17910338 TI - Fine needle aspiration of poorly defined indurated and well-defined breast lesions: a cytopathologic comparative study. AB - OBJECTIVE: To compare the cytologic findings and diagnoses of breast fine needle aspiration (FNA) samples of well-defined lesions (WDL) with those of poorly defined indurated lesions. STUDY DESIGN: We examined 371 consecutive breast FNA specimens obtained without diagnostic image guidance. Fifty-eight lesions were described by the examining pathologists as PDILs, and the remaining 313 lesions were described as WDLs. RESULTS: Compared with WDLs, PDILs were more likely to yield hypocellular specimens deemed unsatisfactory for diagnostic evaluation (37.9% vs. 14.1%). However, a substantial number of atypical, suspicious for malignancy and malignant cases (12.1%, 5.2%, and 13.8%, respectively) were identified with PDILs. In addition, benign diagnoses were more frequently rendered with aspirates of WDLs, compared with PDILs (47.9% vs. 31.0%). In our study, FNAs of PDILs were more often diagnostic in white women < 49 years of age and in lesions measuring > 2 cm. CONCLUSION: Given the relatively high frequency of malignant, suspicious and atypical lesions detected with PDILs, FNA is a suitable first diagnostic approach for PDILs, especially considering the relatively low cost and simplicity of FNA procedures without diagnostic imaging guidance. PMID- 17910339 TI - New cytologic clues in localized Leishmania lymphadenitis. AB - OBJECTIVE: To describe new cytologic clues to diagnose localized leishmania lymphadenitis (LLL). STUDY DESIGN: The study examined cytologic smears of 170 cases of LLL referred to our department from November 1989 to October 2004. A total of 120 cases were confirmed by detecting Leishman-Donovan (LD) bodies in at least 1 of the cytologic smears and 50 cases, which were histologically confirmed. For comparison we studied cytologic smears of 20 cases of tuberculous lymphadenitis, 20 cases of toxoplasma lymphadenitis and 20 cases of granulomatous lymphadenitis of unspecified causes. RESULTS: Cases were divided into 4 major groups. Cytologic findings in these groups were studied to find highly suggestive clues. Cytologic findings present in most of these groups, but absent or very rare in other granulomatous lymphadenitis, were LD kinetoplasts, plasma cells with different shapes of inclusions and lymphogranular bodies. Rare findings not reported previously were: intraneutrophilic LD bodies, hematoxylin body-like inclusions, fibroblasts, cytoplasmic blebbing and floating parasitophorous vacuoles. CONCLUSION: Despite previous reports emphasizing detecting LD bodies in diagnosing LLL, we present cytologic clues highly suggestive of this self-limited disease when LD bodies cannot be detected or are very few on the smears. PMID- 17910340 TI - Fine needle aspiration cytology of bone tumors. AB - OBJECTIVE: To study the role of fine needle aspiration cytology (FNAC) in the diagnosis of bone tumors and its impact on therapeutic decisions. STUDY DESIGN: A group of 122 cases of bone tumor were evaluated by FNAC. Detailed diagnoses were compared with the available histology. RESULTS: Diagnostic accuracy of FNAC was 90.5% in this study. FNAC could differentiate between various round cell tumors such as Ewing's sarcoma and myeloma, among various giant cell-rich lesions of bone and between the benign and malignant chondroid bone tumors. Some uncommon variants were also correctly diagnosed. In metastatic bone tumors, the source of primary malignancy could not be indicated in the majority (52.9%) because of the poorly differentiated morphology. Osteoid or osteoid-like material was demonstrable in 63.6% cases of osteogenic sarcoma. A case of chondroblastic osteogenic sarcoma that was reported as chondrosarcoma was the only diagnostic error in the study. FNAC obviated the need of open biopsy in 63.8% patients, and therapeutic decisions were made according to the cytologic diagnoses. CONCLUSION: FNAC plays an important role in the early diagnosis of bone tumors by its accuracy, ease of use and rapidity and is helpful in making the therapeutic decisions. PMID- 17910341 TI - Critical role of fine needle aspiration cytology and immunocytochemistry in preoperative diagnosis of pediatric renal tumors. AB - OBJECTIVE: To evaluate accuracy and role of immunocytochemistry (ICC) in cytologic diagnosis of pediatric renal tumors. STUDY DESIGN: Fine needle aspirates from 75 cases of pediatric renal tumors were studied. Radiologic-guided aspirations were performed, with 6-7 smears stained with Papanicolaou and Giemsa stains. Smears were screened without the knowledge of final histologic diagnosis. Subsequently, clinical details, final histology and diagnosis rendered by the original cytologist were noted to judge accuracy of diagnosis by a sensitized cytologist. Five neuroblastomas that entered close differentials for Wilms tumor were also evaluated. ICC studies were also performed after staining. RESULTS: Of 58 Wilms tumors, 5 were misdiagnosed; 3 renal rhabdoid tumors and 1 clear cell sarcoma were missed on cytology. Non-Hodgkin's lymphomas presenting as renal masses were accurately diagnosed on cytology, but primitive neuroectodermal tumors (n = 3) and renal cell carcinomas (n = 2) were not accurately diagnosed. Accuracy rate improved from 65% to 92% on review by a cytologist aware of cytologic features of pediatric renal tumors. CONCLUSION: A good accuracy rate of diagnosis of pediatric renal tumors can be achieved by priming pathologists to typical features of tumors. Immunocytochemistry plays a supportive role in cases with atypical morphology or unusual presentations. PMID- 17910342 TI - Outcomes of women with atypical squamous cells of undetermined significance and high-risk human papillomavirus DNA. AB - OBJECTIVE: To study the risk of high-grade squamous intraepithelial lesion (HSIL) and cervical intraepithelial neoplasia (CIN) 2 or 3 in women with human papillomavirus (HPV)-positive atypical squamous cells of undetermined significance (ASCUS) cytology over a 2-year period using the 2001 Bethesda System and ThinPrep Paps. STUDY DESIGN: In 2002, 846 patients with ThinPrep cervical cytology having an ASCUS interpretation and positive for high-risk HPV DNA were identified. A cohort of 514 (60.8%) patients with follow-up by repeat cytology, cervical biopsy or both was included in the study. Patient age was 12-81 years, with a median of 25 years. RESULTS: There were 291 women (56.6%) with negative status by cytology, HPV testing or biopsy with a median interval of 8.5 months, and an additional 174 patients (33.9%) had persistent ASCUS, positive HPV DNA or low-grade SIL/CIN 1. Finally, 49 patients (9.5%) had CIN 2 or 3, with a median interval of 8.5 months. CONCLUSION: Our study suggests that HSIL or CIN 2 or 3 will be detected in 1 in 10 women with HPV-positive index ASCUS cervical cytology at initial colposcopy or within a 2-year follow-up period. PMID- 17910343 TI - Diagnostic value of the thin-layer, liquid-based Pap test in endometrial cancer: a retrospective study with emphasis on cytomorphologic features. AB - OBJECTIVE: To examine the diagnostic value (sensitivity and specificity of the ThinPrep Pap test in the detection of endometrial cancer and assess the morphologic features of endometrial cancer in ThinPrep tests. STUDY DESIGN: In a retrospective, case-controlled study, we identified 60 Pap slides performed within 12 months of the tissue diagnosis of endometrial carcinoma and 56 comparison slides from patients without known carcinoma. The slides were blindly reviewed by the authors without knowledge of the surgical diagnosis. An independent diagnosis was given for the tests based on 14 diagnostic criteria from the 2001 Bethesda System and 6 additional criteria proposed by the authors. RESULTS: The sensitivity of detecting endometrial carcinoma was 88.3% (95% CI 77.8-94.2%) and specificity was 87.5% (95% CI 76.4-93.8%). The positive likelihood ratio was 7.067 (95% CI 3.513-14.217) and negative likelihood ratio was 0.133 (95% CI 0.066-0.269). Enlarged nuclei and the presence of nucleoli in endometrial cells were the most reliable indicators of endometrial cancer or atypical endometrial cells. CONCLUSION: The ThinPrep Pap test has high sensitivity and specificity in detecting or suggesting the presence of endometrial cancer. Certain cytomorphologic features are helpful in distinguishing benign and malignant endometrial lesions. PMID- 17910344 TI - Comparison of two preparation methods for endocervical evaluation. AB - OBJECTIVE: To assess the validity of SurePath liquid-based preparation method for examination of endocervical brush specimens as a substitute for conventionally prepared cytology methods for evaluating the endocervical canal during colposcopic examination and biopsy. STUDY DESIGN: Paired SurePath liquid-based test slides and conventional smears were obtained using an endocervical brush in a split sample protocol before biopsy at the time of colposcopy. The level of agreement between cytologic results obtained was assessed. Accuracy and operating characteristics were evaluated compared to histologic follow-up. RESULTS: Agreement between cytology results for the methods was excellent. The overall kappa was 0.924 (p = 0.0000). There was exact agreement on interpretation between the methods in 283 of 299 cases (94.6%). Cytohistologic follow-up results correlation were: SurePath liquid-based Pap test results and conventional smear results agreed with histology results in 47.8% and 49.2% of cases, respectively. Allowing for a discrepancy within 1 level of severity of cytologic grade, agreements were 76.6% and 77.2%, respectively. CONCLUSION: This study demonstrates that the SurePath method is equivalent to conventional endocervical brush cytology preparation and performs well for detection of cervical intraepithelial lesions and cancer. SurePath is acceptable for endocervical evaluation as a substitute for endocervical curettage at colposcopic biopsy. PMID- 17910345 TI - A human papillomavirus testing system in women with abnormal Pap results: a comparison study with follow-up biopsies. AB - OBJECTIVE: To evaluate the efficacy of INFORM HPV using the SurePath collection method in women whose Pap tests indicated abnormal results. STUDY DESIGN: Ninety two women from the gynecology clinics at The University of Texas M.D. Anderson Cancer Center who had Pap tests and underwent follow-up biopsies were selected for the study. This included 51 women with atypical squamous cells of undetermined significance (ASCUS), 23 women with low-grade squamous intraepithelial lesion (LSIL), 15 women with high-grade squamous intraepithelial lesion (HSIL) and 3 women with negative Pap results. The INFORM HPV, an in situ hybridization assay, testing for oncogenic types of HPV was performed, and the results were compared with follow-up biopsies. RESULTS: The positive rate of the INFORM HPV increased with higher grades of cytology diagnoses. The sensitivity of the INFORM HPV testing for predicting high-grade cervical intraepithelial neoplasia (CIN 2/3) also increased with higher grades of cytology diagnoses. A negative predictive value (NPV) of 94.9% and a specificity of 80.4% for predicting CIN 2/3 were observed in the ASCUS group. CONCLUSION: Using SurePath Pap specimens, the INFORM HPV lacks sufficient sensitivity and NPV for predicting CIN 2/3 in women with ASCUS. Therefore, use of the test as a triage tool is limited. PMID- 17910346 TI - P16(INK4a) immunocytochemistry in liquid-based cytology samples in equivocal Pap smears: added value in management of women with equivocal Pap smear. AB - OBJECTIVE: To test whether p1l6(INK4a) immunocytochemistry (ICC) in liquid-based cytology (LBC) is useful with colposcopy in abnormal Pap smears. STUDY DESIGN: A series of 248 women with abnormal Pap smear were analyzed for oncogenic (HR) human papillomavirus (HPV) types using the Hybrid Capture II assay and for p16(INK4a) expression using ICC on cervical samples in PreservCyt liquid media. Colposcopic and loop electrosurgical excision procedure (LEEP) cone biopsy were the gold standard. RESULTS: p16(INK4a) ICC did best as predictor of high-grade squamous intraepithelial lesion, with OR 12.18 (2.72-54.57) (p = 0.0001), showing 88.2% sensitivity (SE), 61.9% specificity (SP), 14.6% positive predictive value (PPV) and 98.6% negative predictive value (NPV). In sorting discrepant cases, p16(INK4a) ICC results in 100% SE and 100% NPV in detecting cervical intraepithelial neoplasia (CIN) 2 lesions among Pap+/biopsy- women. In atypical squamous cells undetermined significance (ASCUS) cytology, adding p16(INK4a) ICC improves specificity of colposcopy from 27.3% to 81.8% and PPV from 42.8% to 71.4%. Best performance is obtained with p16(INK4a) ICC and colposcopy: 83.3% SE, 81.8% SP, 71.4% PPV and 90.0% NPV. CONCLUSION p16(INK4a) is useful in sorting clinically relevant discrepant cases, and p16(INK4a) ICC significantly improves SP and PPV of colposcopy in management of ASCUS cytology. PMID- 17910347 TI - Role of exfoliative cytology in diagnosis of laryngeal tumors. AB - OBJECTIVE: To evaluate the value of cytologic examination in the diagnosis of laryngeal tumors. STUDY DESIGN: A series of 100 patients with suspected tumorous lesions of the larynx were included in the study. The material for cytologic analysis was obtained by use of a modified instrument, that is, a slightly concave discoid knife, during direct microlaryngoscopy, followed by biopsy for paraffin and frozen section histopathology. The findings obtained by cytology and frozen section histopathology were compared with the findings of paraffin section histopathology. RESULTS: Cytology showed an 83.9% sensitivity, 100% specificity and 90% accuracy compared with histopathologic findings (p < 0.05; chi2 = 7.7). Frozen section pathohistology vs. paraffin section histopathology showed an 80.6% sensitivity, 97.4% specificity and 87% accuracy (p < 0.05; chi2 = 8.1). CONCLUSION: Cytologic examination showed higher accuracy compared to frozen section histopathology, also giving an insight into the type of laryngeal pathology in the case of precancerous lesions. The use of cytology and frozen section histopathology during direct microlaryngoscopy improves clinical diagnostic accuracy. PMID- 17910348 TI - Clinicopathologic significance of eosinophilic pleural effusions in a population with a high prevalence of tuberculosis and cancer. AB - OBJECTIVE: To reveal the diagnostic and prognostic significance of eosinophilic pleural effusions (EPEs) and assess their clinical implications. STUDY DESIGN: Seventy EPEs from 60 patients among 697 consecutive pleural effusions were investigated from 1996-2005 at Kocaeli University Hospital, Kocaeli, Turkey. Koss and Light's criteria were applied in the analysis, which comprised macroscopic, cytopathologic, biochemical and microbiologic examinations. RESULTS: Overall, cancerous underlying conditions were diagnosed in 22 patients (13 malignant and 9 paramalignant), 36.7% of EPEs. Benign causes were found in 43.3% (26 of 60) of the patients. Twelve pleural effusions (20.0%) were idiopathic. The comparison of pleural fluid and peripheral blood findings disclosed no significant difference among the various subgroups. CONCLUSION: EPE could be associated with inflammatory, benign, cancerous and paramalignant conditions. A closer search for a definite causes is warranted in the setting of EPEs, especially in populations with a high prevalence of tuberculosis and malignancy, such as in Kocaeli, Turkey, an industrial city in a developing country. PMID- 17910349 TI - Analysis of urine cytology tests in 120 paired cases. AB - OBJECTIVE: To evaluate the smear quality and diagnostic accuracy of ThinPrep processing in comparison to conventional Cytospin technique for urinary cytology. STUDY DESIGN: ThinPrep and Cytospin techniques were retrospectively evaluated by 2 observers in a double-blinded, randomized fashion. Each quality parameter was scored using a semi-quantitative score of 1-3. Diagnostic accuracy indices were calculated with biopsy histology as the gold standard. RESULTS: Quality of cellular distribution and cell preservation were better with Cytospin preparations, whereas ThinPrep smears were superior in terms of stain distribution and cleaner slide background. However, the only significant differences observed were in cellular distribution and a clean background (p < 0.05). Sensitivity and positive and negative predictive values were higher with Cytospin than the ThinPrep technique (90.0%, 94.7% and 71.4% vs. 80.0%, 94.1% and 55.6%, respectively). Conversely, the specificity of both techniques was comparable. CONCLUSION: The Cytospin smears were of better quality than those prepared by the ThinPrep technique. Although both techniques resulted in similar diagnostic accuracies in negative cases, the ThinPrep preparations were not found to be superior to Cytospin smears in diagnosing positive urinary cytology. PMID- 17910350 TI - Utility of toluidine blue staining and brush biopsy in precancerous and cancerous oral lesions. AB - OBJECTIVE: To evaluate the usefulness of toluidine blue and brush biopsy in precancerous oral lesions and squamous cell carcinoma. STUDY DESIGN: The study was conducted at Moti Lal Nehru Medical College, Allahabad, India. Ninety-six patients with suspicious oral lesions who attended the outpatient clinics of otorhinolaryngology were screened with in vivo toluidine blue staining and oral brush biopsy. RESULTS: Oral brush biopsy showed high specificity and sensitivity. Toluidine blue staining was highly sensitive and moderately specific for malignant lesions but less sensitive for premalignant lesions. CONCLUSIONS: Early detection of oral carcinoma is possible even at the precancerous stages by using noninvasive, painless and outpatient procedures, such as in vivo toluidine blue staining and brush biopsy. PMID- 17910351 TI - Fine needle aspiration cytologic diagnosis of erythema nodosum leprosum: a case report. AB - BACKGROUND: Erythema nodosum leprosum (ENL), the type 2 lepra reaction occurring in lepromatous or borderline lepromatous leprosy, presents clinically with acute manifestations that compel the patient to seek medical attention. Recognition and timely management of these patients is critical in order to avoid permanent disability. Fine needle aspiration cytology (FNAC) is a simple, effective tool that aids in correct diagnosis and management of ENL. CASE: A 30-year-old woman presented with history of fever, reddening of the face, and multiple raised, reddish, painful swellings of the bilateral forearms and legs for 7 days. One year previously, she was diagnosed and treated for lepromatous leprosy with type 2 reaction. After a thorough clinical examination a diagnosis of ENL was made. FNA smears from the forearm swellings showed pus-like material with intact and degenerated polymorphonuclear leukocytes and many foamy macrophages with strong granular acid-fast bacillus (AFB) positivity. A cytologic diagnosis of ENL was given, which was confirmed on histopathologic examination of skin biopsy. CONCLUSION: Cytologic features such as a large number of intact and degenerated neutrophils with foamy macrophages and strong granular AFB positivity, in an appropriate clinical background, allows a confident diagnosis of ENL. PMID- 17910352 TI - Role of fine needle aspiration cytology in detection of microfilariae: report of 2 cases. AB - BACKGROUND: Filariasis is a major public health problem in developing countries, and the diagnosis is conventionally made by demonstrating microfilariae in the peripheral blood smear. However, microfilariae have been incidentally detected in fine needle aspirates of various lesions in clinically unsuspected cases of filariasis with absence of microfilariae in the peripheral blood. CASES: In case 1, a 21-year-old woman presented with multiple left axillary lymphadenopathy of 3 months' duration. In case 2, a 32-year-old woman presented with a thyroid nodule of 7 months' duration. Fine needle aspiration smears from both cases showed sheathed microfilariae of Wuchereria bancrofti. In both cases, microfilariae could not be demonstrated in the peripheral blood smears and the blood eosinophil counts were within normal limits. The histopathologic examination showed neither microfilariae nor adult worm. CONCLUSION: Although microfilariae in cytologic material are considered incidental findings, these cases illustrate the value of routine fine needle aspiration cytology in the detection of asymptomatic and clinically unsuspected cases of bancroftian filariasis. Absence of microfilariae in the peripheral blood does not exdude filarial infection. PMID- 17910353 TI - Epithelial myoepithelial carcinoma of the parotid gland with malignant ductal and myoepithelial components arising in a pleomorphic adenoma: a case report with cytologic, histologic and immunohistochemical correlation. AB - BACKGROUND: To describe the cytologic, histologic and immunohistochemical findings of a case of epithelial myoepithelial carcinoma (EMC) arising from a pleomorphic adenoma (PA) of the parotid with both malignant epithelial and myoepithelial components. CASE: A 29-year-old female presented with a 1.5 x 1.5 cm, palpable mass of the left parotid of 7-8 months' duration with recent enlargement and pain. Fine needle aspiration biopsy (FNAB) revealed biphasic epithelial (small cell) and myoepithelial (large/clear cell) clusters arranged in a pseudopapillary and trabecular pattern with abundant hyaline material with many naked nuclei, together with areas typical of pleomorphic adenoma (PA) was noted. The cytology was reported as salivary gland neoplasm, "suggestive of adenoid cystic carcinoma, less likely pleomorphic adenoma." The mass was excised and histologically reported as "pleomorphic adenoma, with focal invasion of one resected margin." Four months later the tumor recurred, and FNAB showed almost the same cytologic features as did the previous aspirate. Due to early recurrence, previous histologic sections were reviewed, and typical areas of a biphasic pattern of EMC with atypicality and mitosis of both components was found. The final diagnosis was EMC ex PA. CONCLUSION: Although previous reports mention the difficulties in diagnosing EMC and differentiation from the more common salivary gland neoplasms such as PA, we like to emphasize the cytologic confusion that results when the tumors coexist. PMID- 17910355 TI - Primary pulmonary malignant peripheral nerve sheath tumor: a case report. AB - BACKGROUND: Although the histologic features of malignant peripheral nerve sheath tumor (MPNST) have been described, the cytologic features of primary pulmonary MPNST have not been reported in the literature. CASE: We report a case of primary pulmonary MPNST in a 78-year-old man. Follow-up computed tomography of colon cancer, renal cancer, penile cancer and gingival cancer revealed a nodular lesion, 12 mm in diameter, in the right upper lobe of the lung. In frozen section, a diagnosis of malignant neoplasm, not otherwise specified, was rendered for the imprinting specimen and histologic specimen. Imprinting specimens were composed of small cellular aggregates and discohesive neoplastic cells with obvious malignant features. Histologically, spindle cells with pleomorphic nuclei arranged infascicular patterns and multinucleated tumor giant cells were also observed. More than 25 mitotic figures were observed per 10 high-power fields. Tumor cells were positive only for vimentin and S-100, and the Ki-67 labeling index was 10%. Clinical and imaging investigation failed to identify an alternative primary site. We histologically diagnosed this case as primary pulmonary MPNST. CONCLUSION: MPNST has a varied cytomorphology with frank nuclear atypia showing no definite differentiation. Multinucleated neoplastic giant cells with immunopositivity for S-100 may permit more accurate diagnosis of MPNST. PMID- 17910354 TI - Cytologic diagnosis of mastocytosis by fine needle aspiration biopsy: a case report. AB - BACKGROUND: Mastocytosis is an abnormal proliferation of mast cells and their subsequent accumulation in various organs. Diagnosis of mast cell disease relies on proper identification of abnormal mast cells. CASE: A 55-year-old man presented with a history of fever for several months, associated with night sweats, involuntary 20lb weight loss, progressive fatigue, weakness, worsening abdominal distention, shortness of breath, and diffuse lymphadenopathy. Physical examination and computed tomography (CT) showed hepatosplenomegaly, massive ascites, and generalized lymphadenopathy. Bone marrow biopsy with immunohistochemistry (ICH) studies revealed mastocytosis. CT-guided fine needle aspiration biopsy (FNAB) of the retroperitoneal lymphadenopathy was performed. The smears were cellular for a mixed population of mature plasma cells, eosinophils, left-shifted granular and lymphoid cells, and abundant abnormal mast cells. The mast cells had round to oval lobulated nuclei, some of which were binucleated or eccentrically located, with coarse, evenly distributed chromatin. Abundant pale cytoplasm contained numerous metachromatic granules. IHC studies and flow cytometry confirmed the cytologic diagnosis of mastocytosis. CONCLUSION: This case highlights the cytologic features of mastocytosis in FNA specimens. IHC stains and flow cytometry are helpful to confirm the cytologic diagnosis. To the best of our knowledge, this is the second case that describes the cytologic characteristics of mastocytosis. PMID- 17910356 TI - Anaplastic carcinoma of the thyroid with chondroblastoma features mimicking papillary carcinoma: a case report. AB - BACKGROUND: An unusual case of anaplastic carcinoma of the thyroid arising from a metastatic focus of papillary carcinoma. CASE: The tumor affected a 69-year-old woman with a history of total thyroidectomy for papillary thyroid carcinoma 4 years previously. She presented with a rapidly enlarging neck mass that histologically simulated chondroblastoma. A small, embedded focus of residual follicular variant of papillary carcinoma was present. The patient died of disease 3 months later. CONCLUSION: This "chondroblastoma" variant of anaplastic thyroid carcinoma has not been reported to date. PMID- 17910357 TI - Fine needle aspiration cytology in recurrent amelanotic melanoma: a case report. AB - BACKGROUND: Amelanotic melanoma can mimic a wide variety of epithelial and nonepithelial malignant tumors. Varied cytomorphology of melanoma has been described on exfoliative and fine needle aspiration cytology (FNAC). We report a case of recurrent amelanotic melanoma to highlight its varied cytomorphologic features, which may cause diagnostic problems on cytologic and on histologic examinations. CASE: A 63-year-old male presented with nodular swellings in the right anterior chest wall, right axilla and back. A nodule in the chest had been excised 6 months earlier. Clinically, the lesion was interpreted as recurrent soft tissue sarcoma. FNAC revealed malignant cells with highly varied morphology with plasmacytoid and pleomorphic malignant cells with occasional fibrocollagenous tissue strands showing adherent neoplastic cells. A cytologic diagnosis of pleomorphic malignant tumor was suggested, and the original histologic slides were reviewed; they showed a striking alveolar pattern that vaguely resembled an alveolar rhabdomyosarcoma. However, on immunohistochemistry, the tumor cells were S-100 and melan-A positive and desmin negative. A final diagnosis of amelanotic melanoma was made. CONCLUSION: Awareness of the highly varied cytomorphology of amelanotic melanoma minimizes the diagnostic difficulty on fine needle aspiration smears. Suitable immunohistochemical markers are of great value in difficult situations. PMID- 17910358 TI - Cytoplasmic processes as a diagnostic aid in Langerhans cell histiocytosis. PMID- 17910359 TI - Cytomorphology of basaloid (basal cell) carcinoma of the lung. PMID- 17910360 TI - Environmental nanotechnology: a near-term opportunity assessment. PMID- 17910362 TI - Use of chemical coagulants to control fouling potential for wastewater membrane bioreactor processes. AB - Chemical coagulation with ferric chloride, alum, and an organic polymer were used to control the fouling potential of mixed liquors for submerged membrane bioreactor (MBR) processes in treating municipal wastewater. Their filterability was evaluated using a submerged hollow fiber ultrafiltration apparatus operated in constant permeate flux mode. The collected transmembrane pressures over filtration time were used to calculate the membrane fouling rates. The results showed that coagulation pretreatment can reduce fouling rates when MBRs were operated above the critical flux. Even though coagulation with the organic polymer formed larger mixed liquor suspended solids particles and had shorter time-to-filtration than those with ferric chloride and alum, the filterability for membrane filtration were similar, indicating that the membrane fouling in MBR systems was mainly controlled by the concentration of smaller colloidal particles. PMID- 17910361 TI - Activated sludge inhibition by chemical stressors--a comprehensive study. AB - The effects of shock loads of 1-chloro-2,4-dinitrobenzene (CDNB); cadmium; 1 octanol; 2,4-dinitrophenol (DNP); weakly complexed cyanide; pH 5, 9, and 11; and high ammonia levels on activated sludge biomass growth, respiration rate, flocculation, chemical oxygen demand removal, dewaterability, and settleability were studied. For all chemical shocks, except ammonia and pH, concentrations that caused 15, 25, and 50% respiration inhibition were used to provide a single pulse shock to sequencing batch reactor systems containing a nitrifying or non nitrifying biomass. Cadmium and pH 11 shocks were most detrimental to all processes, followed by CDNB. The DNP and cyanide primarily affected respiration, while pH 5, pH 9, octanol, and ammonia did not affect the treatment process to a significant extent. A chemical source-process effect matrix is provided, which we believe will aid in the development of methods that prevent and/or attenuate the effects of toxic shock loads on activated sludge systems. PMID- 17910363 TI - Natural occurrence of metallothioneinlike proteins in liver tissues of four fish species from the northeast Mediterranean Sea. AB - Fish with different ecological needs were captured from the northeast Mediterranean Sea to determine metallothioneinlike proteins in their livers. Heat treated liver samples were run on a column packed with Sephadex G-75 (Sigma, Germany) for subsequent determination of metals, sulfhydryl, and absorbance at 254 and 280 nm. Liver cytosols of Sparus auratus and Mullus barbatus had three heat-stable protein peaks, whereas Mugil cephalus and Atherina hepsetus had two. The third protein peak, containing metallothionein-like proteins (5000 to 6000 Da), was present in all fish. Only high- and low-molecular-weight proteins contained sulfhydryl and metals. Zinc was the most abundant metal in the metallothioneinlike proteins, whereas cadmium and lead were not detected. The differences in the characteristics of metal-binding proteins among fish indicate that the biological characteristics of fish species should be carefully taken into account concerning natural monitoring studies. PMID- 17910364 TI - Effect of cold-temperature shock on nitrification. AB - Nitrification is known as the most temperature-sensitive step among the biological processes in wastewater treatment. The purpose of this study was to investigate the effect of temperature on nitrification, in the case of a sharp decrease of temperature, and to compare this effect with that of a gradual temperature decrease. It was found that a sudden temperature decrease affected nitrification much more than predicted. The immediate decrease of temperature by 10 degrees C led to a 20% larger decrease of specific nitrification rate than predicted by the temperature correction factor of 1.072. The change of nitrification rate resulting from a gradual temperature decrease was modeled correctly with the current default temperature correction factor of 1.072. It was concluded that the correction factor actually can be applied to a gradual temperature-change situation; however, in the case of a sudden temperature decrease, measures need to be taken to avoid nitrifier washout. PMID- 17910365 TI - Free synthetic and natural estrogen hormones in influent and effluent of three municipal wastewater treatment plants. AB - Three municipal wastewater treatment plants (WWTPs) in southeastern Pennsylvania were sampled to determine the presence and concentrations of 12 natural and synthetic estrogen hormones in the wastewater influent and effluent. The target estrogens were 17alpha-estradiol, estrone, estriol, equilin, 17alpha dihydroequilin, 17beta-estradiol, 17alpha-ethinyl estradiol, gestodene, norgestrel, levonorgestrel, medrogestone, and trimegestone. One WWTP uses a biofilm reactor (packed-bed trickling filter),and the other two use suspended growth media (continuously stirred activated sludge reactor and sequential batch reactor). Estrone was detected in all the three plants; estriol and estradiol were detected at two WWTPs; and 17 alpha-dihydroequilin and 17 alpha-ethinyl estradiol were detected at one WWTP. The concentration of estrogens in the influent and effluent of the three treatment plants ranged from 1.2 to 259 ng/L and 0.5 to 49 ng/L, respectively. The percentage removal of estrogens from the aqueous phase ranged from 41 to 99%, except in the case of 17alpha dihydroequilin; the removal of 17alpha-dihydroequilin was negligible. The suspended-growth media systems showed higher removal efficiencies for estrogens than the biofilm system. The analytical method uses a Varian C-18 solid-phase extraction (Varian Inc., Palo Alto, California), followed by a derivatization with bis(trimethylsilyl)trifluoroacetamide. The detection limits for the estrogen compounds ranged from 0.1 to 10 ng/L using a sample size of 1 L. The method recoveries ranged from 71 to 120%, and the relative standard deviation ranged from 6 to 14% for all the hormones. PMID- 17910366 TI - Hydrogen production by anaerobic microbial communities exposed to repeated heat treatments. AB - Biological hydrogen production by anaerobic mixed communities was studied in laboratory-scale bioreactors using sucrose as the substrate. A bioreactor in which a fraction of the return sludge was exposed to repeated heat treatments performed better than a control bioreactor without repeated heat treatment of return sludge and produced a yield of 2.15 moles of hydrogen per mole of sucrose, with 50% hydrogen in the biogas. Terminal restriction fragment length polymorphism analysis showed that two different Clostridium groups (comprised of one or more species) were dominant during hydrogen production. The relative abundance of two other non-Clostridium groups increased during periods of decreased hydrogen production. The first group consisted of Bifidobacterium thermophilum, and the second group included one or more of Bacillus, Melissococcus, Spirochaeta, and Spiroplasma spp. PMID- 17910368 TI - Reduction of photoreactivation with the combined UV/peracetic acid process or by delayed exposure to visible light. AB - Photoreactivation of microorganisms following UV inactivation is a well-known, but complex, phenomenon. It is affected by several factors, including UV fluence, wavelength, light intensity, and exposure time to photoreactivating light. The effect on photoreactivation of a combined peracetic acid (PAA)/UV process has not been investigated. Accordingly, this study compared the degree of photoreactivation, under both sunlight and artificial lights, following UV and combined PAA/UV inactivation of fecal coliforms. Effluent samples from the Montreal Wastewater Treatment Plant (MWTP) (Quebec, Canada) were exposed, for 3 hours, to both low- and high-intensity artificial lights and sunlight. All resulted in similar photoreactivation levels. However, average photoreactivation for UV-treated wastewater samples was 1.2 logs, compared with 0.1 log for the combined PAA/UV treatment. Hence, the use of PAA in combination with UV can significantly reduce the potential for photoreactivation. To simulate the photoreactivation conditions of the MWTP effluent (which passes through a 4-km outfall tunnel with approximately 3 hours detention time), UV-treated samples were kept in the dark for 3 hours before photoreactivating light exposure. After this period, photoreactivation levels were close to zero. Hence, the effects of photoreactivation may be diminished by use of a combined disinfection scheme and/or by delaying exposure of the disinfected wastewater to light. PMID- 17910367 TI - Effect of operational parameters on the removal of particulate chemical oxygen demand in the activated sludge process. AB - The removal of particulate material in the aeration basin of the activated sludge process is mainly attributed to bioflocculation and hydrolysis of particulate substrate. The bioflocculation process in the aeration tank of the activated sludge process occurs only under favorable conditions in the system, and several common operational parameters affect its performance. The principal objective of this research was to observe the effect of mixed liquor suspended solids, solids retention time (SRT), and extracellular polymer substances on the removal of particulate substrate by bioflocculation. A first-order particulate removal expression, based on flocculation, accurately described the removal rates for supernatant suspended solids and colloidal chemical oxygen demand. Based on the results presented in this investigation, a mixed liquor concentration of approximately 2200 mg/L, an SRT of at least 3 days, and a contact time of 30 minutes are needed for relatively complete removal of the particulate substrate in a plug-flow reactor. PMID- 17910369 TI - Biosorption characteristics of copper (II), chromium (III), nickel (II), and lead (II) from aqueous solutions by Chara sp. and Cladophora sp. AB - The aim of this research was to expose individual removals of copper, chromium, nickel, and lead from aqueous solutions via biosorption using nonliving algae species, Chara sp. and Cladophora sp. Optimum pH values for biosorption of copper (II), chromium (III), nickel (II), and lead (II) from aqueous solutions were determined to be 6, 7, 7, and 3 for Cladophora sp. and 5, 3, 5, and 4 for Chara sp. respectively. Maximum adsorption capacities of Chara sp. [10.54 for chromium (III) and 61.72 for lead (II)] and Cladophora sp. [6.59 for chromium (III) and 16.75 and 23.25 for lead (II)] for chromium (III) and lead (II) are similar. On the other hand, copper (II) and nickel (II) biosorption capacity of Cladophora sp. [14.28 for copper (II) and 16.75 for nickel (II)] is greater than Chara sp. [6.506 for copper (II) and 11.76 for nickel (II)]. Significantly high correlation coefficients indicated for the Langmuir adsorption isotherm models can be used to describe the equilibrium behavior of copper, chromium, nickel, and lead adsorption onto Cladophora sp. and Chara sp. PMID- 17910370 TI - Operating parameters for high nitrite accumulation during nitrification in a rotating biological nitrifying contactor. AB - Incomplete nitrification was studied in a completely and partially submerged rotating biological contactor (RBC). In a partially submerged RBC without additional aeration, 50 to 90% nitrite accumulation (alpha) was achieved at rotation speeds (omega) of 2 to 18 min(-1). In a completely submerged RBC operating during 80 days, a higher alpha of 96% was achieved at omega = 2 min( 1). Incomplete nitrification in a completely submerged RBC at oxygen concentrations of 1.5 to 6.8 mg O2/L indicated that the mass transfer of oxygen is rate-limiting. Modeling of the completely submerged RBC predicts that the oxygen profile will not penetrate the biofilm more than 30 microm, thereby strongly limiting the nitrite-oxidizer growth and causing high nitrite accumulation. Molecular analysis (i.e., fluorescence in situ hybridization) indicated that the nitrite-oxidizers are superficially located (<200 microm) and that the ammonia-oxidizers comprise up to approximately 800 microm of the biofilm. PMID- 17910371 TI - Preconcentration of trace arsenite and arsenate with titanium dioxide nanoparticles and subsequent determination by silver diethyldithiocarbamate spectrophotometric method. AB - A novel method of preconcentration of trace arsenite and arsenate by using titanium dioxide nanoparticles as adsorbent was described. The concentrations of preconcentrated arsenite and arsenate were determined by a silver diethyldithiocarbamate spectrophotometric method without desorption. Batch adsorption experiments were carried out as a function of the pH, contact time, amount of titanium dioxide nanoparticles, and solution volume. In the pH range 5 to 6, adsorption rates of arsenite and arsenate were higher than 98%. The calibration coefficient was 0.9991, and the linear range was 0 to 100 microg/L. The developed method was precise, with the relative standard deviation <5% at concentration level of 10 microg/L, with a detection limit (3sigma, n=6) of 0.44 microg/L. The accuracy of the method for total arsenic was validated by standard reference materials (SRM 3103a) (National Institute of Standards and Technology, Gaithersburg, Maryland). The method was also applied to the analysis of arsenite and arsenate in natural water samples to verify the accuracy. The recovery values remained in a narrow range, from 95 to 103%. PMID- 17910372 TI - Breakthrough adsorption study of migratory nickel in fine-grained soil. AB - The present study was conducted to evaluate the breakthrough curve for nickel adsorption in fine-grained soil from a nearby ash pond site of a thermal power plant. Nickel was found to be the major polluting solute in the ash sluicing wastewater. The adsorption of nickel by vertical soil column batch test and horizontal migration test was carried out in the laboratory. Field investigation was conducted also, by installing test wells around the ash pond site. Experimental results showed a good adsorptive capacity of soil for nickel ions. The breakthrough curves showed a reasonable fitting with a one-dimensional mathematical model. The breakthrough curves yielded from field test results showed good agreement with a two-dimensional mathematical model. PMID- 17910373 TI - Characteristics of electrolysis, ozonation, and their combination process on treatment of municipal wastewater. AB - The characteristics of municipal wastewater treatment by electrolysis, ozonation, and combination processes of electrolysis and aeration using three gaseous species (nitrogen [N2], oxygen [O2], and ozone [O3]) were discussed in this research using ruthenium oxide (RuO2)-coated titanium anodes and stainless-steel (SUS304) cathodes. Electrolysis and electrolysis with nitrogen aeration were characterized by a rapid decrease in 5-day biochemical oxygen demand (BODs) and total nitrogen and a slow decrease in chemical oxygen demand (COD). In contrast, ozonation, electrolysis with oxygen aeration, and electrolysis with ozone aeration were characterized by transformation of persistent organic matter to biodegradable matter and preservation of total nitrogen. The best energy efficiency in removing BOD5 and total nitrogen was demonstrated by electrolysis, as a result of direct anodic oxidation and indirect oxidation with free chlorine produced from the chloride ion (Cl-) at the anodes. However, electrolysis with ozone aeration was found to be superior to the other processes, in terms of its energy efficiency in removing COD and its ability to remove COD completely, as a result of hydroxyl radical (*OH) production via cathodic reduction of ozone. PMID- 17910374 TI - Incomplete oxidation of ethylenediaminetetraacetic acid in chemical oxygen demand analysis. AB - Ethylenediaminetetraacetic acid (EDTA) was found to incompletely oxidize in chemical oxygen demand (COD) analysis, leading to incorrect COD values for water samples containing relatively large amounts of EDTA. The degree of oxidation depended on the oxidant used, its concentration, and the length of digestion. The COD concentrations measured using COD vials with a potassium dichromate concentration of 0.10 N (after dilution by sample and sulfuric acid) were near theoretical oxygen demand values. However, COD measured with dichromate concentrations of 0.010 N and 0.0022 N were 30 to 40% lower than theoretical oxygen demand values. Similarly, lower COD values were observed with manganic sulfate as oxidant at 0.011 N. Extended digestion yielded somewhat higher COD values, suggesting incomplete and slower oxidation of EDTA, as a result of lower oxidant concentrations. For wastewater in which EDTA is a large fraction of COD, accurate COD measurement may not be achieved with methods using dichromate concentrations less than 0.1 N. PMID- 17910375 TI - Investigation of pathogenic Escherichia coli and microbial pathogens in pulp and paper mill biosolids. AB - Biosolids produced from pulp and paper mill wastewater treatment have excellent properties as soil conditioners, but often contain high levels of Escherichia coli. E. coli are commonly used as indicators of fecal contamination and health hazard; therefore, their presence in biosolids causes concern and has lead to restrictions in land-spreading. The objectives of this study were to determine the following: (1) if E. coli from the biosolids of a wastewater-free pulp and paper mill were enteric pathogens, and (2) if other waterborne microbial pathogens were present. E. coli were screened for heat-labile and heat-stable enterotoxin and verocytotoxin virulence genes using a polymerase chain reaction. Ten isolates were also screened for invasion-associated locus and invasion plasmid antigen H genes. None of the 120 isolates carried these genes. Tests for seven other microbial pathogens were negative. Effluents and biosolids from this mill do not contain common microbial pathogens and are unlikely to pose a health hazard. PMID- 17910376 TI - Social and demographic factors related to sleep duration. PMID- 17910377 TI - Sleep and the developing brain. PMID- 17910378 TI - Moving towards individualized performance models. PMID- 17910380 TI - American time use survey: sleep time and its relationship to waking activities. AB - STUDY OBJECTIVES: To gain some insight into how various behavioral (lifestyle) factors influence sleep duration, by investigation of the relationship of sleep time to waking activities using the American Time Use Survey (ATUS). DESIGN: Cross-sectional data from ATUS, an annual telephone survey of a population sample of US citizens who are interviewed regarding how they spent their time during a 24-hour period between 04:00 on the previous day and 04:00 on the interview day. PARTICIPANTS: Data were pooled from the 2003, 2004, and 2005 ATUS databases involving N=47,731 respondents older than 14 years of age. INTERVENTIONS: N/A. RESULTS: Adjusted multiple linear regression models showed that the largest reciprocal relationship to sleep was found for work time, followed by travel time, which included commute time. Only shorter than average sleepers (<7.5 h) spent more time socializing, relaxing, and engaging in leisure activities, while both short (<5.5 h) and long sleepers (> or =8.5 h) watched more TV than the average sleeper. The extent to which sleep time was exchanged for waking activities was also shown to depend on age and gender. Sleep time was minimal while work time was maximal in the age group 45-54 yr, and sleep time increased both with lower and higher age. CONCLUSIONS: Work time, travel time, and time for socializing, relaxing, and leisure are the primary activities reciprocally related to sleep time among Americans. These activities may be confounding the frequently observed association between short and long sleep on one hand and morbidity and mortality on the other hand and should be controlled for in future studies. PMID- 17910381 TI - Racial differences in self-reports of sleep duration in a population-based study. AB - STUDY OBJECTIVES: Racial and ethnic differences in sleep duration are not well understood. Research shows that short (< or =6 hours) and long (> or =9 hours) sleepers have higher mortality risks than mid-range sleepers. We investigated whether sleep duration varies by racial and ethnic characteristics and if some of these associations may be explained by residential context. DESIGN: Cross sectional National Health Interview Survey. SETTING: Non-institutionalized adults living in the United States in 1990. PARTICIPANTS: 32,749 people aged 18 years or older. MEASUREMENT AND RESULTS: We estimate a multinomial logistic regression that predicts short, mid-range, and long sleep duration; including covariates for race/ethnicity, among other demographic, health, and neighborhood characteristics. Black respondents had an increased risk of being short and long sleepers (OR=1.41, 95% CI=1.27-1.57 and OR=1.62, 95% CI=1.40-1.88, respectively) relative to white respondents. Hispanics (excluding Mexican Americans) and non Hispanic "Others" were also associated with increased risk of short sleeping (OR=1.26, 95% CI= 1.07-1.49 and OR=1.35, 95% CI= 1.11-1.64, respectively). Living in an inner city was associated with increased risk of short sleeping and reduced risk of long sleeping, compared to non-urban areas. Some of the higher risk of short sleeping among blacks can be explained by higher prevalence of blacks living in the inner city. CONCLUSIONS: Blacks and other racial minorities are more likely to have sleep durations that are associated with increased mortality. The results are consistent with the hypothesis that unhealthy sleep patterns among minorities may contribute to health differentials. PMID- 17910382 TI - Nighttime sleep, Chinese afternoon nap, and mortality in the elderly. AB - STUDY OBJECTIVES: Although many epidemiologic studies have shown that both short and long nighttime sleep durations are associated with increased mortality in the general population, limited data have been reported for older persons, especially those taking afternoon nap. Data from a prospective cohort study of the elderly in Taiwan were used to examine the relationship among nighttime sleep, Chinese afternoon nap, and mortality. DESIGN: Prospective cohort study. SETTING: General population. PARTICIPANTS: A nationally representative sample of 3079 Taiwanese community residents aged 64 and over was studied, using reported sleep related information collected in 1993 and subsequent 10-year mortality data. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Cox proportional hazards models, separated by sex, were computed to estimate mortality hazard ratios in relation to nighttime sleep duration and afternoon nap duration, adjusting for potential confounders. Compared to older adults sleeping 7-7.9 hours at night, those with longer sleeping time (> or = 10 hours in males and > or = 8 hours in females) had a significantly higher risk of total mortality. Afternoon nap alone was not associated with total mortality. When nighttime sleep duration and afternoon nap duration were considered together by adding the interaction term in the model or stratifying sleep hours and nap duration, the effect of afternoon nap on mortality risk remained insignificant. CONCLUSIONS: Longer nighttime sleep duration increases mortality risk in older adults. Chinese afternoon nap is not an independent predictor of mortality. There is no significant benefit or harm of practicing afternoon nap in addition to the regular night sleep on elderly mortality. PMID- 17910383 TI - Daily siesta, cardiovascular risk factors, and measures of subclinical atherosclerosis: results of the Heinz Nixdorf Recall Study. AB - BACKGROUND: Several studies have assessed the association between siesta and cardiovascular outcomes. Concern exists that confounding might have distorted these results and contributed to discrepancies among them. This report examines the association between siesta habits and cardiovascular risk factors, including sleep disturbances at night, depressed mood, and measures of subclinical atherosclerosis such as coronary calcium score and ankle brachial index. METHODS: The baseline examination of 4,797 participants aged 45-74 years included interviews, physical examinations, laboratory tests, and electron beam computed tomography. We compared the baseline prevalence of depressed mood, nighttime sleep disturbances, and health status in 3 categories of siesta habits: irregular or no siestas; daily short siestas (1 hour or less); and daily long siestas (>1 hour). We also characterized cardiovascular risk factor distributions in the 3 siesta groups and conducted a sensitivity analysis of the potential for confounding by these factors in studies of incident cardiovascular disease. RESULTS: Depressed mood and poor self-perceived health status at baseline had positive associations with the age-standardized prevalence of daily long siestas among both men and women. Daily takers of long siestas had a considerably higher prevalence of cardiovascular risk factors in both sexes and appreciably worse measures of subclinical atherosclerosis in men only, in comparison with either of the other siesta groups. Daily long siestas had positive associations with prevalence of several cardiovascular risk factors and measures of subclinical atherosclerosis. CONCLUSIONS: If uncontrolled, these associations could produce appreciable confounding in studies of siesta habits and incidence of cardiovascular events. PMID- 17910384 TI - Sleep duration and sleep complaints and risk of myocardial infarction in middle aged men and women from the general population: the MONICA/KORA Augsburg cohort study. AB - STUDY OBJECTIVES: To examine gender-specific associations between sleep duration and sleep complaints and incident myocardial infarction (MI). DESIGN: Cohort study. SETTING: A representative population sample of middle-aged subjects in Germany. PARTICIPANTS: The study was based on 3508 men and 3388 women (aged 45 to 74 years) who participated in one of the 3 MONICA (Monitoring trends and determinants on cardiovascular diseases) Augsburg surveys between 1984 and 1995, who were free of MI and angina pectoris at baseline and were followed up until 2002. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: A total of 295 cases of incident MI among men and 85 among women occurred during a mean follow-up period of 10.1 years. Compared with women sleeping 8 hours, the multivariable adjusted hazard ratio (HR) of MI among women sleeping < or =5 hours was 2.98 (95% CI, 1.48 6.03), and among women sleeping > or =9 hours 1.40 (95% CI, 0.74-2.64); the corresponding HRs among men were 1.13 (95% CI, 0.66-1.92) and 1.07 (95% CI, 0.75 1.53). In multivariable analysis the relative risk of an incident MI for men and women with difficulties maintaining sleep was 1.12 (95% CI, 0.84-1.48) and 1.53 (95% CI, 0.99-2.37), respectively, and for men and women with difficulties initiating sleep the relative risk was 1.16 (95% CI, 0.82-1.63) and 1.30 (95% CI, 0.81-2.06), respectively. CONCLUSIONS: Modest associations between short sleep duration and difficulties maintaining sleep and incident MI were seen in middle aged women but not men from the general population. PMID- 17910385 TI - Optimization of biomathematical model predictions for cognitive performance impairment in individuals: accounting for unknown traits and uncertain states in homeostatic and circadian processes. AB - Current biomathematical models of fatigue and performance do not accurately predict cognitive performance for individuals with a priori unknown degrees of trait vulnerability to sleep loss, do not predict performance reliably when initial conditions are uncertain, and do not yield statistically valid estimates of prediction accuracy. These limitations diminish their usefulness for predicting the performance of individuals in operational environments. To overcome these 3 limitations, a novel modeling approach was developed, based on the expansion of a statistical technique called Bayesian forecasting. The expanded Bayesian forecasting procedure was implemented in the two-process model of sleep regulation, which has been used to predict performance on the basis of the combination of a sleep homeostatic process and a circadian process. Employing the two-process model with the Bayesian forecasting procedure to predict performance for individual subjects in the face of unknown traits and uncertain states entailed subject-specific optimization of 3 trait parameters (homeostatic build-up rate, circadian amplitude, and basal performance level) and 2 initial state parameters (initial homeostatic state and circadian phase angle). Prior information about the distribution of the trait parameters in the population at large was extracted from psychomotor vigilance test (PVT) performance measurements in 10 subjects who had participated in a laboratory experiment with 88 h of total sleep deprivation. The PVT performance data of 3 additional subjects in this experiment were set aside beforehand for use in prospective computer simulations. The simulations involved updating the subject-specific model parameters every time the next performance measurement became available, and then predicting performance 24 h ahead. Comparison of the predictions to the subjects' actual data revealed that as more data became available for the individuals at hand, the performance predictions became increasingly more accurate and had progressively smaller 95% confidence intervals, as the model parameters converged efficiently to those that best characterized each individual. Even when more challenging simulations were run (mimicking a change in the initial homeostatic state; simulating the data to be sparse), the predictions were still considerably more accurate than would have been achieved by the two-process model alone. Although the work described here is still limited to periods of consolidated wakefulness with stable circadian rhythms, the results obtained thus far indicate that the Bayesian forecasting procedure can successfully overcome some of the major outstanding challenges for biomathematical prediction of cognitive performance in operational settings. PMID- 17910387 TI - Is passive smoking associated with sleep disturbance among pregnant women? AB - STUDY OBJECTIVE: Pregnant women suffer from sleep disturbance, which may be aggravated by passive smoking. In this study we investigated the effects of passive smoking on sleep disturbance during pregnancy. DESIGN: Two cross sectional questionnaire surveys conducted in 2002 and 2006. SETTING: Clinical institutions specializing in obstetrics and gynecology that participated in the nationwide surveys: 260 in the 2002 survey and 344 in the 2006 survey. PARTICIPANTS: 16,396 and 19,386 pregnant women in Japan surveyed in 2002 and 2006, respectively. INTERVENTION: N/A. MEASUREMENTS AND RESULTS: Pregnant women exposed to passive smoking were likely to have sleep disturbances, such as subjective insufficient sleep, difficulty in initiating sleep, short sleep duration, and snoring loudly/breathing uncomfortably. Smoking pregnant women had the same sleep disturbances and also experienced excessive daytime sleepiness and early morning awakening. The prevalence of 5 types of sleep disturbance (insufficient sleep, difficulty in initiating sleep, short sleep duration, excessive daytime sleepiness, and snoring loudly/breathing uncomfortably) among nonsmokers with environmental tobacco smoke showed a mean value intermediate between that of active smokers and that of nonsmokers without environmental tobacco smoke. CONCLUSION: Passive smoking is independently associated with increased sleep disturbance during pregnancy. PMID- 17910386 TI - Elevated inflammatory markers in response to prolonged sleep restriction are associated with increased pain experience in healthy volunteers. AB - CONTEXT: Sleep disturbances, pain, and inflammation co-occur in various medical conditions, but their interrelationships are poorly understood. OBJECTIVE: We investigated the effects of reduced sleep duration (by approximately 50%) to 4 h/night across 10 days, on peripherally circulating inflammatory mediators. In addition, we tested the prediction that degree of inflammation is quantitatively related to the extent to which pain is increased in response to prolonged sleep restriction. DESIGN: Randomized, 16 day controlled in-laboratory study conducted in GCRC. METHODS: Eighteen volunteers were randomly assigned to either 12 days of sleeping 8 h/night or 4 h/night. Participants rated mood and pain symptoms throughout experimental days. Urine was collected and blood was drawn frequently on the baseline day and after the 10th experimental day for 25 hours. OUTCOME MEASURES: Levels of plasma interleukin (IL)-6, serum C-reactive protein (CRP), plasma soluble tumor necrosis factor receptor p55 (sTNF-R p55), urinary levels of prostaglandin (PG) metabolites D2 and E2, subjective assessment of pain and tiredness-fatigue. RESULTS: IL-6 levels were elevated in the 4-h sleep condition over the 8-h sleep condition (P <0.05). CRP levels showed the same trend as IL-6, but did not differ significantly between groups (P = 0.11). Levels of sTNF-R p55 were unchanged in both groups. PG E2 and 11beta-F2alpha metabolite increased in 4 h sleepers, but did not differ significantly from the 8-h sleepers. Elevated IL-6 levels were strongly associated with increased pain ratings in response to sleep restriction (r = 0.67, P <0.01), and this association could not be explained by elevations in tiredness-fatigue. CONCLUSION: Insufficient sleep quantity may facilitate and/or exacerbate pain through elevations of IL-6. In disorders where sleep disturbances are common, insufficient sleep quantity itself may establish and maintain its co-occurrence with pain and increased inflammation. PMID- 17910388 TI - Dream-associated behaviors affecting pregnant and postpartum women. AB - STUDY OBJECTIVES: Evaluate the prevalence and phenomenology of dream-associated behaviors affecting pregnant and postpartum mothers. Episodes consist of anxious dreams and nightmares about the new infant that are accompanied by complex behaviors (motor activity, speaking, expressing emotion). DESIGN: Three-group design (postpartum, pregnant, null gravida), self-report, and repeated measures. SETTING: Pregnancy and postpartum groups: completion of questionnaires in hospital room within 48 hours of giving birth and home telephone interviews; null gravida group: completion of questionnaires and interview in person or by telephone. PARTICIPANTS: Two hundred seventy-three women in 3 groups: postpartum: n = 202 (mean age = 29.7 +/- 4.94 years; 95 primiparas, 107 multiparas); pregnant: n = 50 (mean age = 31.1 +/- 5.44 years); null gravida: n = 21 (mean age = 28.5 +/- 6.34 years). INTERVENTIONS: Subjects completed questionnaires about pregnancy and birth factors, personality, and sleep and participated in interviews concerning the prevalence of recent infant dreams and nightmares, associated behaviors, anxiety, depression, and other psychopathologic factors. MEASUREMENTS AND RESULTS: Most women in all groups recalled dreams (88%-91%). Postpartum and pregnant women recalled infant dreams and nightmares with equal prevalence, but more postpartum women reported they contained anxiety (75%) and the infant in peril (73%) than did pregnant women (59%, P < 0.05 and 42%, P < 0.0001). More postpartum (63%) than pregnant (40%) women reported dream associated behaviors (P < 0.01), but neither group differed from null gravida women (56%). This was due to different distributions over groups of the behavior subtypes. Motor activity was present in twice as many postpartum (57%) as pregnant (24%) or null gravida (25%) women (all P < 0.0001). Expressing emotion was more prevalent among null gravida (56%) than postpartum women (27%) (P < 0.05) but was not different from pregnant women (37%). Speaking was equally prevalent among the 3 groups (12%-19%). Behaviors were associated with nightmares, dream anxiety and, among postpartum women, post-awakening anxiety (41%), confusion (51%), and a need to check on the infant (60%). Primiparas and multiparas differed in dream and nightmare recall but not in prevalence of dream associated behaviors. CONCLUSION: The prevalent occurrence of pregnancy and postpartum infant dreams and associated behaviors may reflect the pervasive emotional influence of maternal concerns or changes instigated by severe sleep disruption, rapid eye movement sleep deprivation, and altered hormone levels. PMID- 17910389 TI - Gender differences in morbidity and health care utilization among adult obstructive sleep apnea patients. AB - STUDY OBJECTIVE: To explore gender differences in morbidity and total health care utilization 5 years prior to diagnosis of obstructive sleep apnea (OSA). DESIGN: Case-control study; patients were recruited between January 2001 and April 2003. SETTING: Two university-affiliated sleep laboratories. PATIENTS: 289 women (22-81 years) with OSA were matched with 289 men with OSA for age, body mass index (BMI), and apnea-hypopnea index (AHI). All OSA patients were matched 1:1 with healthy controls by age, geographic area, and primary physician. MEASUREMENTS AND RESULTS: Women with OSA compared to men with OSA have lower perceived health status and Functional Outcomes of Sleep Questionnaire score (54.5% vs. 28.4%, P <0.05 and 67.5+/-21.4 vs. 76+/-20.1, P <0.05, respectively). Compared to men with OSA, women with OSA have higher risk of hypothyroidism (OR 4.7; 95% CI, 2.3-10) and arthropathy (OR 1.6, 95% CI, 1.1-2.2) and lower risk for CVD (OR 0.7; 95% CI, 0.5-0.91). Compared to controls, both women and men with OSA had 1.8 times higher 5-year total costs (P <0.0001). Compared to men with OSA, expenditures for women with OSA are 1.3 times higher (P <0.0001). The multiple logistic regression (adjusting for BMI, AHI) revealed that age (OR 1.04; 95% CI, 1.01-1.07), antipsychotic and anxiolytic drugs (OR 2.3; 95% CI, 1.2-4.4), and asthma (OR 2.4; 95% CI, 1.1-5.6) are independent determinants for "most costly" OSA women. CONCLUSION: Compared to men with similar OSA severity, women are heavier users of health care resources. Low FOSQ score and poor perceived health status in addition to overuse of psychoactive drugs are associated with high health care utilization among women with OSA. PMID- 17910390 TI - Behavioral correlates of sleep-disordered breathing in older women. AB - STUDY OBJECTIVES: To examine the association between SDB and subjective measures of daytime sleepiness, sleep quality, and sleep related quality of life in a large cohort of primarily community-dwelling older women, specifically considering the relative importance of sleep duration in mediating these associations. DESIGN: Cross-sectional. The functional outcome measures of interest were daytime sleepiness (using the Epworth Sleepiness Scale, ESS), sleep related symptoms (Pittsburgh Sleep Quality Index, PSQI), and sleep related quality of life (Functional Outcomes of Sleep Questionnaire, FOSQ). ANOVA and regression analyses examined the association between SDB severity (measured by indices of breathing disturbances and overnight oxygen saturation) and sleep time (by actigraphy) and these outcome measures. Regression models were adjusted for age, body mass index (BMI), and a medical comorbidity index. We specifically explored whether associations with indices of SDB were mediated by sleep deprivation by adjusting models for actigraphy-determined average total sleep time (TST) during the night. SETTING: Community-based sample examined in home and outpatient settings. PARTICIPANTS: 461 surviving older women from the multicenter Study of Osteoporotic Fractures were examined during Visit 8 from 2002-03. All participants underwent in-home overnight polysomnography for one night and wrist actigraphy for a minimum of 3 24-h periods and completed the above functional outcomes questionnaires. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Participants were aged 82.9 +/- 3.5 (mean +/- SD) years, had BMI of 27.9 +/- 5.1 kg/m2, and had an apnea-hypopnea index (AHI) of 15.7 +/- 15.1. AHI and TST demonstrated a weak correlation (r = -0.15). ESS score individually demonstrated a modest association with AHI, oxygen desaturation, and TST. The association of ESS score and AHI--but not oxygen desaturation-was attenuated to some extent by adjustment for TST. PSQI and FOSQ scores were not associated with measures of SDB severity or TST. CONCLUSIONS: After adjustment for TST, SDB severity in community dwelling older women was not independently associated with self-reported daytime sleepiness, although there may be a modest association that is mediated through reduced TST. In older women, SDB severity was not associated with indices of sleep related symptoms or sleep related quality of life. PMID- 17910391 TI - Nocturnal hypoxia exposure with simulated altitude for 14 days does not significantly alter working memory or vigilance in humans. AB - STUDY OBJECTIVES: To assess the effect of 2 weeks of nocturnal hypoxia exposure using simulated altitude on attention and working memory in healthy adult humans. DESIGN: Prospective experimental physiological assessment. SETTING: General Clinical Research Center. PARTICIPANTS: Eleven healthy, nonsmoking, subjects (7 men, 4 women). The subjects had a mean age of 27 +/- 1.5 years and body mass index of 23 +/- 0.9 kg/m2. INTERVENTIONS: Subjects were exposed to 9 hours of continuous hypoxia from 2200 to 0700 hours in an altitude tent. Acclimatization was accomplished by graded increases in "altitude" over 3 nights (7700, 10,000 and 13,000 feet), followed by 13,000 feet for 13 consecutive days (FIO2 0.13). MEASUREMENTS AND RESULTS: Polysomnography that included airflow measurements with a nasal cannula were done at baseline and during 3 time points across the protocol (nights 3, 7, and 14). Attention (10-minute Psychomotor Vigilance Task) and working memory (10-minute verbal 2-back) were assessed at baseline and on day 4, 8, 9, and 15. Nocturnal hypoxia was documented using endpoints of minimum oxygen saturation, oxygen desaturation index, and percentage of total sleep time under 90% and 80%. Total sleep time was reduced, stage 1 sleep was increased, and both obstructive and nonobstructive respiratory events were induced by altitude exposure. There was no difference in subjective mood, attention, or working memory. CONCLUSIONS: Two weeks of nocturnal continuous hypoxia in an altitude tent did not induce subjective sleepiness or impair objective vigilance and working memory. Caution is recommended in the extrapolation to humans the effects of hypoxia in animal models. PMID- 17910392 TI - Discrepancy between subjective symptomatology and objective neuropsychological performance in insomnia. AB - STUDY OBJECTIVES: While daytime impairment is a defining feature of primary insomnia (PI), prior research using objective measures has not yielded clear and reliable evidence of global or specific deficits. In this investigation subjective and neuropsychological measures of daytime impairment were concurrently evaluated in subjects with primary insomnia (PIs) and in healthy good sleeper subjects (GSs). The goals for the study were to assess (1) whether PIs differ from GSs on subjective and/or objective measures and (2) the extent to which subjective and objective measures provide discordant information. DESIGN: Subjects were evaluated on multiple self-report measures of sleep and daytime performance and were administered a comprehensive set of neuropsychological tests. SETTING: The University of Rochester Sleep and Neurophysiology Research Laboratory (Rochester, NY). PATIENTS OR PARTICIPANTS: Forty-nine subjects (32 PIs and 17 GSs). Seventy-one percent of the sample was female; average age 39 +/- 11 yrs. RESULTS: Overall, PIs reported worse sleep, diminished activity levels, and a greater number and severity of daytime complaints. However, PIs did not show deficits on neuropsychological tests. Additionally, neuropsychological measures were not associated with severity of daytime complaints. Objectively measured sleep was found to be associated with performance (motor speed), while prospective and objective sleep measures were associated with level of daytime complaint. CONCLUSIONS: The discrepancy between subjective daytime complaints and objective performance in individuals with insomnia is common, but poorly understood. This discordance may suggest that daytime impairment corresponds less to "output" and more to attentional bias or to the realistic appraisal that "effort" is required to maintain normal performance. PMID- 17910393 TI - Associations between sleep duration patterns and behavioral/cognitive functioning at school entry. AB - OBJECTIVE: The aim of the study was to investigate the associations between longitudinal sleep duration patterns and behavioral/cognitive functioning at school entry. DESIGN, SETTING, AND PARTICIPANTS: Hyperactivity-impulsivity (HI), inattention, and daytime sleepiness scores were measured by questionnaire at 6 years of age in a sample of births from 1997 to 1998 in a Canadian province (N=1492). The Peabody Picture Vocabulary Test--Revised (PPVT-R) was administered at 5 years of age and the Block Design subtest (WISC-III) was administered at 6 years of age. Sleep duration was reported yearly by the children's mothers from age 2.5 to 6 years. A group-based semi-parametric mixture model was used to estimate developmental patterns of sleep duration. The relationships between sleep duration patterns and both behavioral items and neurodevelopmental tasks were tested using weighted multivariate logistic regression models to control for potentially confounding psychosocial factors. RESULTS: Four sleep duration patterns were identified: short persistent (6.0%), short increasing (4.8%),10 hour persistent (50.3%), and 11-hour persistent (38.9%). The association of short sleep duration patterns with high HI scores (P=0.001), low PPVT-R performance (P=0.002), and low Block Design subtest performance (P=0.004) remained significant after adjusting for potentially confounding variables. CONCLUSIONS: Shortened sleep duration, especially before the age of 41 months, is associated with externalizing problems such as HI and lower cognitive performance on neurodevelopmental tests. Results highlight the importance of giving a child the opportunity to sleep at least 10 hours per night throughout early childhood. PMID- 17910394 TI - Adolescent use of mobile phones for calling and for sending text messages after lights out: results from a prospective cohort study with a one-year follow-up. AB - OBJECTIVE: To assess the prevalence of the use of mobile phones by adolescents after lights out and its relationship to tiredness levels after one year. DESIGN: Prospective cohort study with self-reports and follow up questionnaire after one year. SETTING: Second- and fifth-year secondary school children in 15 schools in Flanders, Belgium. PARTICIPANTS: 1656 school children; 52.1% boys. Average age was 13.7 years (SD: 0.68) in the youngest group and 16.9 years (SD: 0.83) in the oldest group at baseline. MAIN OUTCOME MEASURES: Self-reported tiredness. RESULTS: Only 38% of the subjects never used their mobile phone after lights out. Multinomial logistic regression showed that using the mobile phone less than once a month increased the odds of being very tired one year later by 1.8 (95% CI 1.2 2.8). Those who used it less than once a week were 2.2 times more likely to be very tired (95% CI 1.4-3.5). Using it about once a week increased the odds by 3.3 (95% CI 1.9-5.7) and those who used it more than once a week were 5.1 times more likely to be very tired (95% CI 2.5-10.4). Overall 35% of the cases of being very tired were attributed to the use of the mobile phone. Use of the phone right after lights out increased the odds of being very tired by 2.2 (95% CI 1.4-3.4); between 00:00 and 03:00 the odds were 3.9 times higher (95% CI 2.1-7.1), and in those who used it at any time of the night the odds were 3.3 times higher (95% CI 1.8-6.0). CONCLUSION: Mobile phone use after lights out is very prevalent among adolescents. Its use is related to increased levels of tiredness. There is no safe dose and no safe time for using the mobile phone for text messaging or for calling after lights out. PMID- 17910395 TI - Delayed sleep phase disorder in temporal isolation. AB - STUDY OBJECTIVES: This study sought to characterize sleep and the circadian rhythm of body core temperature of an individual with delayed sleep phase disorder (DSPD) in the absence of temporal cues and social entrainment and to compare those measures to age-matched normal control subjects studied under identical conditions. DESIGN: Polysomnography and body temperature were recorded continuously for 4 days in entrained conditions, followed immediately by 17 days in a "free-running" environment. SETTING: Temporal isolation facility in the Laboratory of Human Chronobiology, Weill Cornell Medical College. PARTICIPANTS: One individual who met criteria for delayed sleep phase disorder according to the International Classification of Sleep Disorders Diagnostic and Coding Manual (ICSD-2) and 3 age-matched control subjects. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: The DSPD subject had a spontaneous period length (tau) of 25.38 hours compared to an average tau of 24.44 hours for the healthy controls. The DSPD subject also showed an altered phase relationship between sleep/wake and body temperature rhythms, as well as longer sleep latency, poorer sleep efficiency, and altered distribution of slow wave sleep (SWS) within sleep episodes, compared to control subjects. CONCLUSIONS: Delayed sleep phase disorder may be the reflection of an abnormal circadian timing system characterized not only by a long tau, but also by an altered internal phase relationship between the sleep/wake system and the circadian rhythm of body temperature. The latter results in significantly disturbed sleep, even when DSPD patients are permitted to sleep and wake at their preferred times. PMID- 17910396 TI - "A new lachrymal gland with an excretory duct in red and fallow deer" by Johann jacob Harder (1694): English translation and historical perspective. AB - The Harderian gland is an enigmatic orbital gland that has been described for many tetrapods, although a consistent definition of this structure has remained elusive. In particular, an unambiguous distinction between the Harderian gland and the nictitans gland, which may both occur in the anterior aspect of the orbit of mammals, remains problematic. These glands were first distinguished in 1694 by Johann Jacob Harder, a Swiss physician and anatomist. To facilitate a renewed examination of the anatomical and developmental relationships of the anterior orbital glands, we review the historical context of Harder's discovery, and provide Harder's original Latin text as well as an English translation. PMID- 17910397 TI - One gland, two lobes: organogenesis of the "Harderian" and "nictitans" glands of the Chinese muntjac (Muntiacus reevesi) and fallow deer (Dama dama). AB - The nictitans and Harderian glands are enigmatic glands situated in the anterior aspect of the orbit. Traditionally, the nictitans and Harderian glands of mammals have been considered to be two fundamentally distinct glands. However, a consistent, unambiguous distinction between these two glands has remained elusive due to conflicting anatomical and histochemical definitions. The Harderian gland was originally described, and first distinguished from the nictitans gland, in adult deer. We examined the organogenesis and histochemistry of the anterior orbital glandular mass in two species of deer (Muntiacus reevesi and Dama dama) to determine whether it comprises two distinct glands or one bilobed gland. The anterior orbital regions of 30 fetal specimens of both species, along with some adult material, were examined histologically. Four stages of glandular organogenesis were observed. Most notably, both glandular portions developed from the same inception point, but the deep lobe developed faster than the superficial lobe. The common inception point and the relationship of the collecting ducts clearly shows that this is a single glandular mass that differentiates into two lobes rather than two distinct glands. Moreover, although the histochemical profiles of the two lobes differ slightly, both lobes produce lipids, which is further indication that these are not profoundly different glands but part of a single, heterogeneously developed gland. Thus, it is proposed that the terms nictitans and Harderian glands, as separate entities, be discontinued and that the entire gland be referred to as the anterior orbital gland (glandula orbitalis anterior), with superficial and deep lobes (pars superficialis and pars profundus, respectively). PMID- 17910398 TI - The crista supraventricularis in the human heart and its role in the morphogenesis of the septomarginal trabecula. AB - The crista supraventricularis and septomarginal trabecula are common elements of the right ventricle, and determine many hemodynamic phenomena. The morphological analysis of both structures in regard to their mutual relations was the aim of this study. The study was carried out on the material of preserved human hearts- fetuses, children and adults. The size and development of the crista supraventricularis was carefully evaluated. The division of its lower part, and hence the possibilities of development of the septomarginal trabecula, was divided into five types (A, B, C, D and E). The most common was type B, containing two muscular trabeculae. The width of the crista varied 1/5-3/5 of the width of the interventricular septum. On the basis of this study, a conclusion of morphological unity of the septomarginal trabecula and crista supraventricularis was drawn. PMID- 17910399 TI - Atrial natriuretic peptide (ANP)-granules in the guinea pig atrial and auricular cardiocytes: an immunocytochemical and ultrastructural morphometric comparative study. AB - The atrial natriuretic peptide (ANP) is a peptide hormone that is mainly produced in the cardiac atria, where it is stored within granules. It is known that the four regions of the atrial-auricular complex (two atria and two auricles) produce and store ANP in the granules. However, no report has been presented comparing the presence of ANP, and the number and diameter of atrial granules in the atria and auricles. ANP immunoreactivity was detected in cardiocytes from the four regions of the atrial-auricular complex. No differences were observed among the regions. The number of granules was greatest in the right atrium followed by the left atrium and left auricle and right auricle, in this order. The diameter of granules in the cardiocytes was significantly largest in the right atrium and reduced via the left auricle to the left atrium and right auricle. Both the number and diameter of the granules are larger in the right atrium in comparison with the other regions of the atrial-auricular complex, which leads to the supposition that this region is the one that most synthesizes and stores the ANP. PMID- 17910400 TI - Morphometric study of the ascending aorta in human fetuses. AB - The present study was performed on 128 spontaneously aborted human fetuses, aged 15-34 weeks, to compile normative data for ascending aorta dimensions at varying gestational age. Using anatomical dissection, digital-image analysis (system of Leica QWin Pro 16) and statistical analysis (ANOVA, regression analysis) a range of measurements (Length, original and terminal external diameters, volume) for the ascending aorta during gestation was examined. No significant gender differences were found (P > 0.05). The growth curves of the best fit for the plot for each morphometric feature against gestational age were generated. Both the Length and external diameters of the ascending aorta were found to increase in a linear fashion throughout gestation. The Length ranged from 2.63 +/- 0.42 to 10.80 +/- 1.49mm, according to the linear function y = -4.678 + 0.4647x +/- 0.8447 (r = 0.95). The original external diameter ranged from 2.02 +/- 0.26 to 6.84 +/- 0.63 mm, according to the linear model y = -2.103 + 0.2684x +/- 0.3958 (r = 0.97). The terminal external diameter ranged from 1.73 +/- 0.20 to 6.29 +/- 0.52 mm, with accordance to the linear function y = -2.354 + 0.2567x +/- 0.3826 (r = 0.97). The ascending aorta volume ranged from 7.56 +/- 2.65 to 370.99 +/- 105.42 mm3, according to the quadratic function y = 373.1 - 43.38x + 1.30x(2) +/- 24.51 (R2 = 0.89). The growth curves generated from my data might be useful as a reference for fetal echocardiographers in the detection of some congenital cardiovascular abnormalities. PMID- 17910401 TI - Axillary arch in human: common morphology and variety. Definition of "clinical" axillary arch and its classification. AB - In this work the authors summarize the extensive information available concerning the best-known variant muscular structure in the region of the human axilla--the axillary arch. Emphasis has been placed on the common morphology and variety of the axillary arches. From the anatomical descriptions, the authors extract the characteristics of a group of "typical" axillary arches and also noted the descriptions of a group of "unusual" axillary arches found in the recent literature. The axillary arch terminology, incidence in human population, innervation and origin are discussed. The clinical significance of the axillary arch is presented briefly and the need of a new concept for the axillary arch is stated. In conclusion, for the purpose of clinical practice, a new term- "clinical" axillary arch is defined, and its proper clinically oriented classification is presented. PMID- 17910402 TI - A common celiacomesenteric trunk, and a brief review of the literature. AB - The authors report a rare variation, a common celiacomesenteric trunk, which was observed during routine dissection of an 89-year-old Japanese female cadaver in the laboratory of the Anatomy Department. The trunk gave rise to left gastric, common hepatic, splenic and superior mesenteric arteries. The developmental significance of this variation is discussed with a brief review of the literature. PMID- 17910403 TI - Two anatomical autopsy cases of direct communication between a persistent primitive trigeminal artery and an anterior inferior cerebellar artery. AB - The persistent primitive trigeminal artery (PPTA) is the most common persistent carotid-basilar anastomosis. However, morphological findings of the PPTA based on the anatomical autopsy are very scarce. To understand the reason why such a variant artery develops, it is essential to examine the detailed morphology of the PPTA and developmental process of this artery. Here, we present two anatomical autopsy instances of the PPTA (cases 1 and 2). In the first case (78 year-old female; right side), the anterior inferior cerebellar artery (AICA) arose from the internal carotid artery passing medial to the abducens nerve. This artery gave off a small branch communicating to the basilar artery, passed lateral to the trigeminal nerve root, and continued backward to the dorsal surface of the cerebellum. Thus, in this case, the AICA is considered to be branched from the PPTA. In the second case (75-year-old female, left side), the PPTA branched from the internal carotid artery, and passed lateral to the abducens nerve, giving off an artery connecting with the AICA. These communicating arteries between the basilar artery and the AICA, recognized in cases 1 and 2, are considered to be the persistence of the primitive lateral basilovertebral anastomosis during the early embryological period. We propose that the primitive lateral basilovertebral anastomosis forms the arterial network around the trigeminal nerve root, and the AICA develops through this anastomosis. PMID- 17910404 TI - Age-related structural and neurochemical changes of the human superior cervical ganglion. AB - The aim of this study was to investigate age-related morphological and neurochemical changes in the human superior cervical ganglion (SCG). Thirty-seven superior sympathetic human cervical ganglia of young, adult, and aged subjects were examined using morphometric analysis, biotin-streptavidin immunohistochemistry for detecting neurofilament, myelin protein, protein gene product 9.5, nerve growth factor receptor p75 in sympathetic neurons and nerve fibers. Morphometric parameters of neurons (area, long and short axis, shape factor of the neuron body, nucleus, cytoplasm, and lipofuscin) were investigated in every sixth serial section of the ganglion. Seven hundred neurons with clearly visible nuclei were measured in each studied group. The present study showed that human SCG of older subjects had larger areas of neuron body, cytoplasm and nucleus, a lower shape factor, an increased amount of lipofuscin, and a greater number of large-size neurons, as compared to SCG obtained from young subjects. Neuronal cytoskeletal alterations manifested themselves through a decreased number of neurofilament-positive neurons were detected in old human SCG. The amount of myelinated fibers decreased with age, although the amount of myelinated fibers in the young and the adult subjects varied from few to a moderate number. PGP 9.5 immunoreactivity varied in different age groups. A marked reduction of nerve growth factor receptor p75 in old human sympathetic neurons was detected. In conclusion, the findings of this study confirm age-related morphological changes in the human SCG. Structural neuronal changes may influence the deterioration of neuronal functional capacity, neuronal plasticity, and regenerative characteristics. PMID- 17910405 TI - Anterior intermeniscal ligament: an ultrastructural study. AB - The aim of the present study was to investigate the detailed histological characteristics of membranous and cord-like anterior intermeniscal ligaments (AIMLs) by transmission electron microscopy (TEM) and light microscopy. Ten biopsies of AIMLs were sampled from 10 knees during total knee arthroplasty procedures. Three of them were membranous and 7 of them were cord-like. They were processed for light and TEM evaluations. Histologically, the findings in the membranous and cord-like ligaments were similar. They consisted of parallel bundles of collagen fibrils and their posterior surfaces were covered by a layer of loose well-vascularized synovial tissue. The subsynovial region consisted of loose connective tissue and was rich in blood vessels and nerve endings. Fibroblasts embedded between parallel-oriented collagen fibrils were the major cell type that we observed. Free nerve endings were squeezed between bundles of collagen fibers. Electron microscopic observations revealed the presence of Ruffini corpuscles. The presence of neural mechanoreceptors in the membranous and cord-like intermeniscal ligaments may contribute to structural and proprioceptional function of the knee. Protection of those ligaments may be valuable in planning and performing meniscal surgeries. PMID- 17910406 TI - The intraindividual agreement of the bone density of the human proximal tibia. AB - The interindividual variability in the biomechanical properties of cadaver bones has remained an unsolved problem in biomechanical investigation procedures. For this reason, it is postulated to use matched bone pairs from the same individual for comparative biomechanical tests. The rationale behind this procedure is based on the assumption that biomechanically similar behaviour is to be expected in an intraindividual rather than an interindividual comparison. Systematic studies confirming this thesis were performed on the human femur. However, investigations regarding the intraindividual properties of the proximal tibial metaphysis with respect to the underlying bone densities, have not yet been performed. In order to verify the hypothesis that matched proximal tibial metaphyses from the same donor imply corresponding bone density values, densitometric measurements (pQCT) were performed in 14 matched cadaver tibias (average age 61 years, 9 men, 5 women) which were fresh-frozen at -40 degrees C after removal. After statistical analysis of the bone density values, five tibial pairs were identified as differing on the basis of missing correlations and the existence of systematic differences within the pairwise data. In other words, only about 2/3 of the data in the random sample available was classified as comparable. As the bone density measured by pQCT technique significantly correlates with the biomechanical properties of the bone, it can be concluded from the test result available that matched human tibiae show no concurring bone density values in 1/3 of cases. Thus the pairing of corpse tibiae does not necessarily imply suitability for comparative biomechanical experiments. PMID- 17910407 TI - Morphometric and ultrastructural analysis of stage-specific effects of Sertoli and spermatogenic cells seen after short-term testosterone treatment in young adult rat testes. AB - The effects of testosterone treatment on spermatogenesis in the rat have been investigated by morphometric and structural analysis at the ultrastructural level in stages VII-IX. The aim has been to characterize the changes in Sertoli and spermatogenic cells to elucidate the mechanism of testosterone effects on spermatogenesis and to test the possibilities of developing male contraceptives. In stage VII, the morphometric parameters of volume and surface area in Sertoli cells (see abbreviations below): and the morphometric parameter of volume in the spermatogenic cells such as V(VPG,T), V(VPC,T), V(VrPT,T) and V(VelPT,T) decreased. In stage VIII, the respective values of Sertoli cells, VSN, and VSN/VSC decreased while SSJ increased, and the respective morphometric parameters in the spermatogenic cells, V(VPG,T), V(VPC,T), and V(VrPT,T) increased. In stage IX, in Sertoli cells VSC, VSN, VSN/VSC, and SSJ remained unchanged. In spermatogenic cells V(VPG,T), V(VPC,T), and V(VrPT,T) increased. Further, in all stages, a close apposition of mitochondria and rough endoplasmic reticulum in basolateral cytoplasm of Sertoli cells suggested active protein synthesis. In elongated spermatids in stage IX the microtubular manchette became disorganized. This disorganization and the unexpected shift after testosterone treatment from decrease in several morphometric parameters in stage VIII to increases in stage IX cannot be explained by alterations in testosterone (T), LH, FSH, and their respective receptors. Therefore, still unknown regulatory factors in spermatogenesis are apparently involved in the developmental interactions between Sertoli and spermatogenic cells. PMID- 17910408 TI - [Educative intervention and development of position and critical reading]. AB - OBJECTIVE: To investigate the professors of technical courses of the area of health, the effects of a promotional educative strategy of the participation in the development of a position prior to the education and of the aptitude for the critical theoretical text reading and information of educative research. MATERIAL AND METHODS: A longitudinal study took place, of intervention. In order to measure the degree of development of a position before the education, it was applied to the instrument Concepts and ideas about education. It consists of 72 statements, organized in duple that expresses two different approaches from education: participative and passive. For the inquiry of the degree of development of critical reading two instruments were applied: 1) Theoretical text reading of education and, 2) Reading information of educative research, constituted both by 120 itemes. The validity and trustworthiness of the three instruments were valued by experts with experience in teaching and educative research. The strategy was implemented through activities in seminary form, which were done twice a week, with a duration of five hours per session, for nine months; within the activities outside the classroom, the student completed reading of a theoretical text and/or report of educative research and the resolution of a reading guide (task). During the activities in a propitious classroom, discussion atmosphere and promoting at any moment the participation of the students; a space for the reflective recovery of their own experience was opened, for the analysis and interchange of ideas and for the critic and self criticism of the main educative practices. The professor intervened when individual participation diminished; he also channeled the discussion, indicating to the enlightening observations and strong arguments of the students. The three instruments were applied to the group of professors of technical courses (n = 10); the initial measurement was applied before initiating the educative strategy and the final measurement at the end of the same plan. The qualification of the instruments and the capture of information were made by a blind technical worker. RESULTS: Subsequent to the educative strategy we observed a statistically significant advance in position -inferred through its main indicator: critical theoretical text consequence- and reading. The advances shown in critical reading for information of educative research were below the critical theoretical text reading. CONCLUSIONS: The development of position before the education and the aptitudes for critical reading of theoretical texts and information of educative research, in professors of technical courses of the area of health, it is possible, if educative atmospheres are created that lead to the participation halfway through by the critic. PMID- 17910409 TI - Nondiagnostic thyroid fine needle aspiration cytology: outcome in surgical treatment. AB - BACKGROUND: Fine-Needle Aspiration (FNA) is the main screening process for distinguishing benign from malignant thyroid nodules. Despite this, by 5-29% of patients, their FNA results are not enough to confirm malign neoplasia, particularly in cases with follicular lesions. The objective of this report is to present the definitive histological results of a group of 41 patients with FNA of Thyroid nodule catalogued as "indeterminate/non diagnostic" sent for surgical treatment. MATERIAL AND METHODS: A retrospective analysis was done on all of the patients who had underwent surgery for thyroid nodule, with a previous diagnosis of "indeterminate/non diagnostic" by FNA. Forty-one patients, three male (7.31%), and 38 female (92.68%), were included in the present study. RESULTS: Fifteen women and one man were positive for malignancies (39.02%). The nodule was bigger than 4 cm in 23 patients in total (56.09%), and of this percentage, 6 were malignant (26.09%). According to age, 24 patients were older than 45 years (58.5%), 8 of whom showed malignant pathology (33.3%). All these variables were non significant. Fifteen of 16 patients had a definitive diagnosis of papillary carcinoma and one follicular carcinoma. CONCLUSIONS: The majority of patients with a diagnosis of "indeterminated/non diagnostic" had benign lesions (60.9%). The usual predictive factors for malignity such as age, sex, size of nodule, did not present a significant support in the differential diagnosis. PMID- 17910411 TI - Mortality predictive indexes in non-critical inpatients. AB - BACKGROUND: Mortality predictive indexes have not been applied to patients in general wards out of the ICU. METHODS: Retrolective study aimed to evaluate the value of mortality prediction indexes in a cohort of 944 non-critical patients. Three indexes were evaluated according to their calibration and discriminative power: the Mortality Probability Model II (MPMII), the Simplified Acute Physiology System II (SAPS II) and the Logistic Organ Dysfunction System (LODS). The bivariate calculation of relative risk (RR) to die was performed relative to the group of patients that had an expected probability to die > 10%, calculated by an index. To evaluate the calibration, data were arranged in descending order using the chi2 goodness-of-fit model. To evaluate discrimination power, ROC curves were used. RESULTS: SAPS II, MPM II and LODS predicted significant risks at levels of P < 0.005, (RR = 6.56, 4.03 and 3.44, respectively). Regarding the calibration, the null hypothesis was accepted only by using SAPS II (P = 0.664). CONCLUSIONS: The three evaluated indexes each had a good discriminative capacity to detect non-critical inpatients with high risk to die. SAPS II was the best index to predict mortality, as determined by both the bivariate and the calibration analysis. There is no reason for not using mortality predictive indexes for non-critical inpatients. PMID- 17910410 TI - Patient and technique survival in continuous ambulatory peritoneal dialysis in a single center of the west of Mexico. AB - INTRODUCTION: In Mexico, CAPD survival has been analyzed in few studies from the center of the country. However, there are concerns that such results may not represent what occurs in other province centers of our country, particularly in our geographical area. AIM: To evaluate the patient and technique survival on CAPD of a single center of the west of Mexico, and compare them with other reported series. DESIGN: Retrospective cohort study. SETTING: Tertiary care, teaching hospital located in Guadalajara, Jalisco. PATIENTS: Patients from our CAPD program (1999-2002) were retrospectively studied. Interventions. Clinical and biochemical variables at the start of dialysis and at the end of the follow up were recorded and considered in the analysis of risk factors. MAIN OUTCOME MEASURES: Endpoints were patient (alive, dead or lost to follow-up) and technique status at the end of the study (June 2002). RESULTS: 49 patients were included. Mean patient survival (+/- SE) was 3.32 +/- 0.22 years (CI 95%: 2.9-3.8 years). Patients in the present study were younger (39 +/- 17yrs), had larger body surface area (1.72 +/- 0.22 m2), lower hematocrit (25.4 +/- 5.2%), albumin (2.6 +/- 0.6g/dL), and cholesterol (173 +/- 44 mg/dL), and higher urea (300 +/- 93 mg/dL) and creatinine (14.9 +/- 5.6 mg/ dL) than those in other Mexican series. In univariate analysis, the following variables were associated (p < 0.05) to mortality: pre-dialysis age and creatinine clearance, and serum albumin and cholesterol at the end of follow-up. In multivariate analysis, only pre-dialysis creatinine clearance (RR 0.66, p = 0.03) and age (RR 1.08, p = 0.005) significantly predicted mortality. Mean technique survival was 2.83 +/- 0.24 years (CI 95%: 2.4-3.3). Pre-dialysis age (p < 0.05), peritonitis rate (p < 0.05), and serum phosphorus at the end of follow-up (p < 0.05) were associated with technique failure in univariate analysis, while in multivariate analysis, only pre-dialysis age (RR 1.07, p = 0.001) and peritonitis rate (RR 481, p < 0.0001) were technique failure predictors. CONCLUSIONS: Patients from this single center of the west of Mexico were younger, had higher body surface area and initiated peritoneal dialysis with a more deteriorated general status than patients reported in other Mexican series; in spite of the latter, patient and technique survival were not different. In our setting, pre-dialysis older age and lower CrCl significantly predicted mortality, while older predialysis age and higher peritonitis rate predicted technique failure. PMID- 17910412 TI - [Cost-effectiveness of the treatment of acute and chronic rhinosinusitis at the IMSS]. AB - INTRODUCTION: Rhinosinusitis is one of the more common diseases encountered in outpatient visits to health care. The objective of this study was to determine the most cost-effective antibiotic treatment for patients with acute (RSA) and chronic rhinosinusitis (RSC) that is available at the Mexican Institute of Social Security (IMSS). METHODS: Cost-effectiveness analysis of RSA and RSC treatment from an institutional perspective. Effectiveness outcome was defined as the percentage of cure. A decision tree with a Bayesian approach included the following therapeutic alternatives: ciprofloxacin, gatifloxacin, trimetoprim/sulfametoxazol (TMP/SMX), amoxicilin/clavulanic acid (AAC) and clindamicin. RESULTS: Treatment for RSA with AAC showed a mean cost per cured patient of $ 878 pesos. The remaining antibiotics had a higher cost per unit of success, and therefore the results showed that AAC was the best alternative considering this criterion. The therapy that showed a larger percentage of cured patients in RSC was clindamicin; however, the therapeutic alternative with the lowest cost per successful unit was the one based on ciprofloxacin, which dominates gatifloxacin and AAC. CONCLUSIONS: The most cost-effective alternative in the antibiotic treatment of patients with RSA was ACC while for RSC it was ciprofloxacin; sensitivity analysis showed the strength of the base study results. PMID- 17910413 TI - [Impact on human health of hormonal additives used in animal production]. AB - The establishment of the impact of environmental compounds or additives with hormone-like activity on human health still requires further investigation, as well as a reexamination of biologic models and experimental methodology employed so far. In 1988, the FAO/WHO Expert Committee on Food Additives Joint with the Federal Drug Administration (FDA) considered that sexual hormone residues usually present in meat do not represent a risk for human consumption. Nevertheless, this resolution seems to be uncertain since the scientific elements employed for this statement may not be adequate. In this review the principal objections to the evidence used to establish the innocuousness of growth promoter hormones are considered. PMID- 17910414 TI - [Non-surgical acute abdomen as a clinical expression of Mediterranean famililal fever]. PMID- 17910415 TI - [Neurotrophic control in the development and function of two endocrine organs: the ovary and the pancreas]. AB - Neurotrophins (NTs) are important for the survival, differentiation and function of sympathetic and sensorial neurons of central and peripheral nervous system. However, similar functions have been described of NTs in non-neural organs. Nerve Growth factor (NGF) participates in the foliculogenesis and ovulation in the ovary, as well as in the islet morphogenesis and insulin secretion of the pancreatic beta cell. The NTs act by binding to two distinct classes of transmembranal receptors: p75 and Trks. Both receptor types lead to activation of intracellular signaling cascades that end with cell survival or apoptosis. In this review different actions of the NTs in the ovarian and the pancreas are described. PMID- 17910416 TI - Anodic electrochemiluminescence of CdTe quantum dots and its energy transfer for detection of catechol derivatives. AB - This work reported for the first time the anodic electrochemiluminescence (ECL) of CdTe quantum dots (QDs) in aqueous system and its analytical application based on the ECL energy transfer to analytes. The CdTe QDs were modified with mercaptopropionic acid to obtain water-soluble QDs and stable and intensive anodic ECL emission with a peak value at +1.17 V (vs Ag/AgCl) in pH 9.3 PBS at an indium tin oxide (ITO) electrode. The ECL emission was demonstrated to involve the participation of superoxide ion produced at the ITO surface, which could inject an electron into the 1Se quantum-confined orbital of CdTe to form QDs anions. The collision between these anions and the oxidation products of QDs led to the formation of the excited state of QDs and ECL emission. The ECL energy transfer from the excited CdTe QDs to quencher produced a novel methodology for detection of catechol derivatives. Using dopamine and L-adrenalin as model analytes, this ECL method showed wide linear ranges from 50 nM to 5 microM and 80 nM to 30 microM for these species. Both ascorbic acid and uric acid, which are common interferences, did not interfere with the detection of catechol derivatives in practical biological samples. PMID- 17910417 TI - A system for LogD screening of 96-well plates using a water-plug aspiration/injection method combined with high-performance liquid chromatography mass spectrometry. AB - A system for screening of the octanol/water distribution coefficient (LogD) using automatic sampling of 96-well plates was developed. The high-speed assay for LogD (HSLogD) screening uses a water-plug aspiration/injection method combined with high-performance liquid chromatography-mass spectrometry (HPLC-MS). The method is useful for LogD analysis of highly hydrophobic compounds, where the concentration of the compound in the octanol phase is much higher than that in the water phase. In the case of LogD analyses, the conventional shake-flask method has been widely used, but it is difficult to increase the throughput of the shake-flask method because the lower water phase is carefully separated by manual separation without contamination of the upper octanol phase. We attempted to develop an automatic sampling method instead of manual separation to increase the throughput of the measurement. In initial attempts at automatic sampling, contamination of the octanol phase occurred when sampling of the water phase was made. This was because the octanol phase entered the sampling needle as it passed through to the lower water phase. This contamination was prevented by taking up a few microliters of water into the needle as a plug before sampling of the water phase (the water-plug aspiration/injection method). LogD values of some common drugs measured using the HSLogD agreed with reported LogD values (0 < LogD < 5). PMID- 17910418 TI - Biochemical basis for enhanced binding of peptide dimers to X-linked inhibitor of apoptosis protein. AB - XIAP (X-linked inhibitor of apoptosis protein) is involved in the mediation of programmed cell death and, therefore, is a target for the development of cancer therapeutics. Peptide mimetics based upon Smac, the natural binding partner of XIAP, and specifically, dimeric peptides, have shown great promise in drug development. In the present work, the basis for enhanced dimer efficacy has been explored. Comparisons are made between the peptide binding site on the BIR3 domain of XIAP alone (residues 238-358) and a less truncated construct that includes both BIR2 and BIR3 domains (residues 151-350). This contingency differentially enhances the binding of dimeric tetrapeptides, potentially by providing additional hydrophobic binding surface. The effect of BIR2 on the BIR3 binding site is sustained, even if the BIR2 binding site is disrupted by mutagenesis, as shown by both a fluorescent competition assay and a polarity sensitive dye, badan. FRET measurements reveal an observed separation of >or=45 A between the BIR2 and BIR3 peptide binding pockets, thereby precluding a direct simultaneous interaction of the dimer molecules with both binding domains. Furthermore, variations in the linker length between dimeric tetrapeptides did not show a predictable trend in binding affinities, suggesting that local concentration effects were also an unlikely explanation for the enhanced dimeric affinities. Taken together, the results suggest that enhanced binding of dimeric peptides likely reflects the increased hydrophobic surface area on or near the BIR3 site and have significant ramifications for the design of therapeutics that target this class of proteins. PMID- 17910419 TI - Enzyme kinetics of muscle glycogen phosphorylase b. AB - Interest in the kinetics of glycogen phosphorylase has recently been renewed by the hypothesis of a glycogen shunt and by the potential of altering phosphorylase to treat type II diabetes. The wealth of data from studies of this enzyme in vitro and the need for a mathematical representation for use in the study of metabolic control systems make this enzyme an ideal subject for a mathematical model. We applied a two-part approach to the analysis of the kinetics of glycogen phosphorylase b (GPb). First, a continuous state model of enzyme-ligand interactions supported the view that two phosphates and four ATP or AMP molecules can bind to the enzyme, a result that agrees with spectroscopic and crystallographic studies. Second, using minimum error estimates from continuous state model fits to published data (that agreed well with reported error), we used a discrete state model of internal molecular events to show that GPb exists in three discrete states (two of which are inactive) and that state transitions are concerted. The results also show that under certain concentrations of substrate and effector, ATP can activate the enzyme, while under other conditions, it can competetively inhibit or noncompetitively inhibit the enzyme. This result is unexpected but is consistent with spectroscopic, crystallographic, and kinetic experiments and can explain several previously unexplained phenomena regarding GPb activity in vivo and in vitro. PMID- 17910420 TI - Substrate specificity of homogeneous monkeypox virus uracil-DNA glycosylase. AB - Weak or nonexistent smallpox immunity in today's human population raises concerns about the possibility of natural or provoked genetic modifications leading to re emergence of variola virus and other poxviruses. Thus, the development of new antiviral strategies aimed at poxvirus infections in humans is a high priority. The DNA repair protein uracil-DNA glycosylase (UNG) is one of the viral enzymes important for poxvirus pathogenesis. Consequently, the inhibition of UNG is a rational therapeutic strategy for infections with poxviruses. Monkeypox virus, which occurs naturally in Africa, can cause a smallpox-like disease in humans. Here, the monkeypox virus UNG (mpUNG) is characterized and compared to vaccinia virus UNG (vUNG) and human UNG (hUNG). The mpUNG protein excises uracil preferentially from single-stranded DNA. Furthermore, mpUNG prefers the U.G pair over the U.A pair and does not excise oxidized bases. Both mpUNG and vUNG viral proteins are strongly inhibited by physiological concentrations of NaCl and MgCl2. Although the two viral DNA repair enzymes have similar substrate specificities, the kcat/KM values of mpUNG are higher than those of vUNG. The mpUNG protein was strongly inhibited by 5-azauracil and to a lesser extent by 4(6)-aminouracil and 5-halogenated uracil analogues, whereas uracil had no effect. To develop antiviral drugs toward mpUNG, we also validated a repair assay using the molecular beacons containing multiple uracil residues. Potential targets and strategies for combating pathogenic orthopoxviruses, including smallpox, are discussed. PMID- 17910422 TI - Eigen and Zundel forms of small protonated water clusters: structures and infrared spectra. AB - The spectral properties of protonated water clusters, especially the difference between Eigen (H3O+) and Zundel (H5O2+) conformers and the difference between their unhydrated and dominant hydrated forms are investigated with the first principles molecular dynamics simulations as well as with the high level ab initio calculations. The vibrational modes of the excess proton in H3O+ are sensitive to the hydration, while those in H5O2+ are sensitive to the messenger atom such as Ar (which was assumed to be weakly bound to the water cluster during acquisitions of experimental spectra). The spectral feature around approximately 2700 cm-1 (experimental value: 2665 cm-1) for the Eigen moiety appears when H3O+ is hydrated. This feature corresponds to the hydrating water interacting with H3O+, so it cannot appear in the Eigen core. Thus, H3O+ alone would be somewhat different from the Eigen forms in water. For the Zundel form (in particular, H5O2+), there have been some differences in spectral features among different experiments as well as between experiments and theory. When an Ar messenger atom is introduced at a specific temperature corresponding to the experimental condition, the calculated vibrational spectra for H5O2+.Ar are in good agreement with the experimental infrared spectra showing the characteristic Zundel frequency at approximately 1770 cm-1. Thus, the effect of hydration, messenger atom Ar, and temperature are crucial to elucidating the nature of vibrational spectra of Eigen and Zundel forms and to assigning the vibrational modes of small protonated water clusters. PMID- 17910423 TI - Solvent and solvent isotope effects on the vibrational cooling dynamics of a DNA base derivative. AB - Vibrational cooling by 9-methyladenine was studied in a series of solvents by femtosecond transient absorption spectroscopy. Signals at UV and near-UV probe wavelengths were assigned to hot ground state population created by ultrafast internal conversion following electronic excitation by a 267 nm pump pulse. A characteristic time for vibrational cooling was determined from bleach recovery signals at 250 nm. This time increases progressively in H2O (2.4 ps), D2O (4.2 ps), methanol (4.5 ps), and acetonitrile (13.1 ps), revealing a pronounced solvent effect on the dissipation of excess vibrational energy. The trend also indicates that the rate of cooling is enhanced in solvents with a dense network of hydrogen bonds. The faster rate of cooling seen in H2O vs D2O is noteworthy in view of the similar hydrogen bonding and macroscopic thermal properties of both liquids. We propose that the solvent isotope effect arises from differences in the rates of solute-solvent vibrational energy transfer. Given the similarities of the vibrational friction spectra of H2O and D2O at low frequencies, the solvent isotope effect may indicate that a considerable portion of the excess energy decays by exciting relatively high frequency (>/=700 cm-1) solvent modes. PMID- 17910421 TI - Side chain and backbone dynamics of phospholamban in phospholipid bilayers utilizing 2H and 15N solid-state NMR spectroscopy. AB - 2H and 15N solid-state NMR spectroscopic techniques were used to investigate both the side chain and backbone dynamics of wild-type phospholamban (WT-PLB) and its phosphorylated form (P-PLB) incorporated into 1-palmitoyl-2-oleoyl-sn glycerophosphocholine (POPC) phospholipid bilayers. 2H NMR spectra of site specific CD3-labeled WT-PLB (at Leu51, Ala24, and Ala15) in POPC bilayers were similar under frozen conditions (-25 degrees C). However, significant differences in the line shapes of the 2H NMR spectra were observed in the liquid crystalline phase at and above 0 degrees C. The 2H NMR spectra indicate that Leu51, located toward the lower end of the transmembrane (TM) helix, shows restricted side chain motion, implying that it is embedded inside the POPC lipid bilayer. Additionally, the line shape of the 2H NMR spectrum of CD3-Ala24 reveals more side chain dynamics, indicating that this residue (located in the upper end of the TM helix) has additional backbone and internal side chain motions. 2H NMR spectra of both WT-PLB and P-PLB with CD3-Ala15 exhibit strong isotropic spectral line shapes. The dynamic isotropic nature of the 2H peak can be attributed to side chain and backbone motions to residues located in an aqueous environment outside the membrane. Also, the spectra of 15N-labeled amide WT-PLB at Leu51 and Leu42 residues showed only a single powder pattern component indicating that these two 15N-labeled residues located in the TM helix are motionally restricted at 25 degrees C. Conversely, 15N-labeled amide WT-PLB at Ala11 located in the cytoplasmic domain showed both powder and isotropic components at 25 degrees C. Upon phosphorylation, the mobile component contribution increases at Ala11. The 2H and 15N NMR data indicate significant backbone motion for the cytoplasmic domain of WT-PLB when compared to the transmembrane section. PMID- 17910424 TI - Modulation of excited-state intramolecular proton transfer by viscosity in protic media. AB - 3-Hydroxyquinolones undergo excited-state intramolecular proton transfer (ESIPT), resulting in a dual emission highly sensitive to H-bonding perturbations. Here, we report on the strong effect of viscosity on the dual emission of 2-(2-thienyl) 3-hydroxyquinolone in protic solvents. An increase in viscosity significantly decreases the formation of the ESIPT product, thus changing dramatically the ratio of the two emission bands. Time-resolved studies suggest the presence of solvated species characterized by decay times close to the solvent relaxation times in viscous media. The intramolecular H bond in this species is probably disrupted by the solvent, and therefore, its ESIPT requires a reorganization of the solvation shell for restoring this intramolecular H bond. Thus, the ESIPT reaction of this dye and its dual emission depend on solvent relaxation rates and, therefore, on viscosity. The present results suggest a new physical principle for the fluorescence ratiometric measurement of local viscosity. PMID- 17910425 TI - Intersystem crossing processes in nonplanar aromatic heterocyclic molecules. AB - A comprehensive study of the photophysical properties of a series of monoaza[5]helicenes is presented on the basis of joint optical spectroscopy and quantum chemistry investigations. The molecules have been characterized by absorption and CW/time-resolved luminescence measurements. All quantities related to spin-orbit-coupling processes, such as intersystem crossing rates and radiative phosphorescence lifetimes, were found to depend strongly on the nitrogen position within the carbon backbone. Density functional theory and semiempirical quantum-chemical methods were used to evaluate the molecular geometries, the characteristics of the excited singlet and triplet states, and the spin-orbit coupling matrix elements. We demonstrate that the magnitude of spin-orbit coupling is directly correlated with the degree of deviation from planarity. The trends from the calculated photophysical quantities, namely, radiative fluorescence and phosphorescence decay rates and intersystem crossing rates, of the mono-aza-helicenes are fully consistent with experiment. PMID- 17910426 TI - Synthesis and biological evaluation of new 2-arylcarbonyl-3 trifluoromethylquinoxaline 1,4-di-N-oxide derivatives and their reduced analogues. AB - As a continuation of our research in the quinoxaline 1,4-di-N-oxide new series of 2-arylcarbonyl-3-trifluoromethylquinoxaline, 1,4-di-N-oxide derivatives have been synthesized and evaluated in a full panel of 60 human tumor cell lines. Selective reductions were carried out on two compounds which allowed us to determine the compound structures by comparison of the 1H NMR spectra. In general, all the di-N oxidized compounds showed good cytotoxic parameters. The best activity was observed in derivatives with electron-withdrawing groups in position 6 or 7 on the quinoxaline ring and in the unsubstituted analogues, whereas loss of one or two oxygens reduced the cytotoxicity. The best five compounds were selected for evaluation for the in vivo hollow fiber assays. In vitro studies reveal that compound 5h efficiently generates reactive oxygen species via redox cycling in the presence of the NADPH/cytochrome P450 enzyme system, providing a plausible molecular mechanism for the observed aerobic cytotoxicity of these quinoxaline N oxides. PMID- 17910427 TI - Rational design of 5'-thiourea-substituted alpha-thymidine analogues as thymidine monophosphate kinase inhibitors capable of inhibiting mycobacterial growth. AB - Recently, thymidine monophosphate kinase (TMPK) emerged as an attractive target for developing inhibitors of Mycobacterium tuberculosis growth. The elucidation of the X-ray structure of TMPK of M. tuberculosis (TMPKmt), as well as the structure of an earlier serendipitously discovered dimeric thymidine inhibitor, laid the foundation for the design of potent and selective TMPKmt inhibitors reported here. Several hits identified within a series of 3'-C-branched thiourea substituted beta-thymidine derivatives inspired us to construct a set of 5' thiourea-substituted alpha-thymidine derivatives characterized by a similar relative orientation of the thymine and arylthiourea moieties. alpha-Thymidine derivative 15, featuring a (3-trifluoromethyl-4-chlorophenyl)thiourea moiety, has a Ki of 0.6 microM and a selectivity index of 600 versus human TMPK. Moreover, it represents the first TMPK inhibitor showing good inhibitory activity on growing M. bovis (MIC99 = 20 microg/mL) and M. tuberculosis (MIC50 = 6.25 microg/mL) strains. PMID- 17910428 TI - Synthesis and monoamine oxidase inhibitory activity of new pyridazine-, pyrimidine- and 1,2,4-triazine-containing tricyclic derivatives. AB - A number of condensed azines, mostly belonging to the families of indeno-fused pyridazines (1), pyrimidines (2, 3), and 1,2,4-triazines (4, 5), were synthesized and evaluated in vitro as monoamine oxidase (MAO) A and B inhibitors. Most of them showed higher inhibition potency toward MAO-B, the most effective one being 3-(3-nitrophenyl)-9H-indeno[1,2-e] [1,2,4]triazin-9-one (4c), which displayed an IC50 value of 80 nM and proved to be 10-fold more potent than its [2,1-e] fusion isomer 5. Replacing the 3-phenyl group of the known indeno[1,2-c]pyridazin-5-one MAO-B inhibitors with a flexible phenoxymethyl group enhanced the inhibitory potency. The inhibition data highlighted the importance of the aza-heterocyclic scaffold in affecting the MAO isoform selectivity. The 3-phenyl derivatives with type 1, 4, and 5 scaffolds were inhibitors of MAO-B with little or no MAO-A effect, whereas 2- or 3-phenyl derivatives of type 2 and 3 pyrimidine-containing fusion isomers inhibited both isoenzymes with a structure-dependent preference toward MAO-A. PMID- 17910429 TI - Synthesis and biological properties of novel 2-aminopyrimidin-4(3H)-ones highly potent against HIV-1 mutant strains. AB - Following the disclosure of dihydro-alkoxy-, dihydro-alkylthio-, and dihydro alkylamino-benzyl-oxopyrimidines (DABOs, S-DABOs, and NH-DABOs) as potent and selective anti-HIV-1 agents belonging to the non-nucleoside reverse transcriptase inhibitor (NNRTI) class, we report here the synthesis and biological evaluation of a novel series of DABOs bearing a N,N-disubstituted amino group or a cyclic amine at the pyrimidine-C2 position, a hydrogen atom or a small alkyl group at C5 and/or at the benzylic position, and the favorable 2,6-difluorobenzyl moiety at the C6 position (F2-N,N-DABOs). The new compounds were highly active up to the subnanomolar level against both wt HIV-1 and the Y181C mutant and at the submicromolar to nanomolar range against the K103N and Y188L mutant strains. Such derivatives were more potent than S-DABOs, NH-DABOs, and nevirapine and efavirenz were chosen as reference drugs. The higher inhibitor adaptability to the HIV-1 RT non-nucleoside binding site (NNBS) may account for the higher inhibitory effect exerted by the new molecules against the mutated RTs. PMID- 17910431 TI - New and original pKa prediction method using grid molecular interaction fields. AB - One of the most important physicochemical properties of a molecule is pKa. It is known that two parameters imperative in ADME profiling, solubility, and lipophilicity are governed by pKa, and receptor binding can be influenced by pKa. Because most drugs are ionized in physiological conditions, pKa is particularly relevant to medicinal chemistry. Despite the numerous advances in high-throughput measurements, in silico determination is still the fastest and cheapest way of obtaining pKa. This paper presents a new original computational method for pKa prediction of organic compounds. Descriptors were generated using the program GRID, and these descriptors are based on molecular interaction fields precomputed on a set of molecular fragments. The new method was developed, trained, and cross validated by using a large and diverse data set of 24 617 pKa values. This paper presents the results for a class of 421 acidic nitrogen compounds (RMSE = 0.41, r2 = 0.97, q2 = 0.87) and for a class of 947 six-membered N-heterocyclic bases (RMSE = 0.60, r2 = 0.93, q2 = 0.85). For external validation 28 novel compounds were selected that covered nine different ionizable groups, and 39 pKa values could be experimentally determined by spectral gradient analysis (SGA). Comparison of experimental pKa with calculated pKa demonstrated that the predictive ability of the method is good (external set, r2 = 0.85, RMSE = 0.90). PMID- 17910433 TI - Strength of the Zn-N coordination bond in zinc porphyrins on the basis of experimental thermochemistry. AB - The compound, 5,10,15,20-tetrakis(4-methoxyphenyl)porphine zinc(II) (ZnTMPP), was prepared, and its thermochemical properties were experimentally established. The standard molar energy of combustion (Delta(c)U degrees m) was determined from oxygen rotating-bomb combustion calorimetry experiments. The standard molar enthalpies of combustion (Delta(c)H degrees m) and formation (Delta(f)H degrees m) were derived. The enthalpy of sublimation (Delta(cr)(g)H degrees m) was determined by Knudsen effusion at high temperatures. With these results, the standard molar enthalpies of formation and atomization (Delta(at)H degrees m) in the gas state were calculated. A summary of the results at T = 298.15 K (p degrees = 0.1 MPa) is shown in Table 1. Using these results and those previously obtained for the free ligand, 5,10,15,20-tetrakis(4-methoxyphenyl)porphine, the mean dissociation enthalpy for the Zn-N coordination bond is obtained as D(Zn-N) = (160 +/- 9) kJ.mol-1. This value is consistent with the results obtained using the same experimental approach in a similar system (5,10,15,20 tetraphenylporphine, TPP/ZnTPP) reported elsewhere. A discussion of the strength for the Zn-N coordination bond is made in terms of the structural and electronic features of the molecules involved. PMID- 17910432 TI - Synthesis, crystal structure, and magnetic properties of hexanuclear [{MnL2}4{Nb(CN)8}2] and Nonanuclear [{MnL2}6{Nb(CN)8}3] heterometallic clusters (L=bpy, phen). AB - A hexanuclear cyano-bridged {MnII4NbIV2} cluster (1) bearing 2,2'-bipyridine (bpy) as the blocking ligand at manganese is obtained from the reaction of cis [MnCl2(bpy)2] and K4[Nb(CN)8]. When the blocking ligand is 1,10-phenanthroline (phen), a nonanuclear cluster {MnII6NbIV3} (2) is obtained. The structure of [{Mn(bpy)2}4{Nb(CN)8}2] has been solved by single-crystal X-ray crystallography, whereas the phen derivative has been confirmed by means of the structure analysis of the corresponding WIV analogue [{Mn(phen)2}6{W(CN)8}3(H2O)2]. Magnetic measurements revealed S=9 and 27/2 spin ground states for these aggregates as a result of antiferromagnetic Nb-Mn interaction with JNb-Mn=-18.1 cm(-1) (1) and 13.6 cm(-1) (2). PMID- 17910434 TI - Hydrothermal synthesis, structural chemistry, and magnetic properties of materials of the MII/triazolate/anion family, where MII = Mn, Fe, and Ni. AB - Hydrothermal chemistry has been exploited in the preparation of a series of manganese(II), iron(II), and nickel(II) triazolate frameworks, [Mn7(trz)8(CH3CO2)4(OH)2].2.5H2O (1.2.5H2O), [Mn5(Htrz)2(SO4)4(OH)2] (2), [Fe5(Htrz)2(SO4)4(OH)2] (3), [Fe3(Htrz)3(HSO4)(SO4)2(OH)].H2O (4.H2O), [Ni3(trz)3(OH)3(H2O)4].5H2O (5.5H2O), and [Ni3(trz)5(OH)].2.5H2O (6.2.5H2O). The materials all exhibit three-dimensional structures, reflecting the tendency of triazole/triazolate ligands to bridge multiple metal sites. A prominent characteristic of the structures is the presence of embedded metal clusters as building blocks: heptanuclear MnII units in 1, pentanuclear MII sites in 2 and 3, and trinuclear MII clusters in 4 and 5. The presence of the pentanuclear and trinuclear clusters of magnetic metal cations in 2-5 is reflected in the unusual magnetic characteristics of these materials, all of which exhibit spin frustration. The compound 5.5H2O reversibly desorbs/sorbs solvent. However, the dehydrated phase does not adsorb methanol, N2, O2, or H2, presumably as a consequence of the highly polar void volume and the narrow channels connecting the larger cavities of the void structure. PMID- 17910435 TI - Synthesis, structure, and magnetic properties of [(CH3CN)5V-O-V(CH3CN)5][BF4]4. AB - The reaction of vanadium(III) acetylacetonate with HBF4 in acetonitrile yields [(CH3CN)5V-O-V(CH3CN)5][BF4]4, a material that serves as a convenient precursor to other [V-O-V]4+ species such as [(bipy)2(CH3CN)V-O-V(CH3CN)(bipy)2][BF4]4 (bipy=2,2'-bipyridine). Single-crystal X-ray diffraction shows that the V-O-V linkage of [(CH3CN)5V-O-V(CH3CN)5]4+ is linear. An Evans method measurement of the solution-phase magnetic susceptibility indicates strong ferromagnetic coupling between the vanadium centers. Magnetic susceptibility (chi) and magnetization (M(H)) data for a powdered sample and for a single crystal oriented with its V-O-V axis parallel to the applied field were measured over 1.8-300 K. The results suggest that the V(III) centers are ferromagnetically coupled with J approximately 72 K (approximately 50 cm(-1)) yielding a ground state with a total spin Stotal=2. Theoretical fit to the M(H) plot for the single crystal yielded g||=2.01+/-0.01 and the zero-field splitting parameter D=0.60+/-0.04 K (0.42+/ 0.03 cm(-1)). EPR measurements at 34 and 101.6 GHz are consistent with the Stotal=2 ground state and yield g||=1.9825, g perpendicular=1.9725 and D=0.57+/ 0.03 K. PMID- 17910436 TI - Self-assembly of ligands designed for the building of a new type of [2 x 2] metallic grid. anion encapsulation and diffusion NMR spectroscopy. AB - The ligands 4,6-bis(pyrazol-1-yl)pyrimidine (bpzpm), 4,6-bis(3,5-dimethylpyrazol 1-yl)pyrimidine (bpz(*)pm), 4,6-bis(4-methylpyrazol-1-yl)pyrimidine (Mebpzpm), and 3,6-bis(3,5-dimethylpyrazol-1-yl)pyridazine (ppdMe) were synthesized and were made to react with Cu(I) centers in the presence of different counteranions. Different [2 x 2] metallic grids were obtained. With ligands bpzpm, bpz*pm, and Mebpzpm, a new type of grid was obtained where the facing ligands were divergent and two counteranions (BF(4-) or PF(6-)) were hosted in the resulting cavities and exhibit C-H...F and anion...pi interactions in the solid state. The presence of methyl groups on the pyrazolyl rings induced several distortions in the structure. In complexes with the ligand ppdMe, there were found two groups of parallel ligands in the grid, and the cavities generated were smaller. The counteranions were situated outside the grid, and the facing ligands exhibited aromatic pi-pi stacking interactions. Anion-pi interactions involving the pyridazine ring were found. The behavior in solution of the new derivatives with a special emphasis on the cation-anion interactions was studied by UV-vis and NMR spectroscopy. Diffusion NMR experiments performed for some complexes allowed us to conclude that weak cation-anion interactions exist in solution, with the counteranions undergoing fast exchange on the diffusion time scale between the free and ion-paired states. PMID- 17910437 TI - Synthesis and characterization of a Mn22 single-molecule magnet and a [Mn22]n single-chain magnet. AB - The reactions of [Mn12O12(O2CEt)16(H2O)4] with phenylphosphinic acid (PhHPO2H) in MeCN and MeCN/CH2Cl2 have led to isolation of [Mn22O12(O2CEt)22(O3PPh)8(H2O)8] (2) and [Mn22O12(O2CEt)20(O3PPh)8(O2PPhH)2(H2O)8]n (3), respectively, both containing PhPO3(2-) groups from in situ oxidation of PhHPO(2)(-). Complex 2 is molecular and consists of two Mn9 subunits linked by four additional Mn atoms. Complex 3 contains almost identical Mn22 units as 2, but they are linked into a one-dimensional chain structure. The Mn22 unit in both compounds is mixed-valence Mn(III)18, Mn(II)4. Solid-state, variable-temperature dc magnetic susceptibility and magnetization measurements were performed on vacuum-dried samples of 2 and 3, indicating dominant antiferromagnetic interactions. A good fit of low-temperature magnetization data for 2 could not be obtained because of problems associated with low-lying excited states, as expected for a high nuclearity complex containing Mn(II) atoms. An approximate fit using only data collected in small applied fields indicated an S = 7 or 8 ground state for 2. Solid-state ac susceptibility data established that the true ground state of 2 is S = 7 and that the connected Mn22 units of 3 are ferromagnetically coupled. Both 2 and 3 displayed weak out-of-phase ac signals indicative of slow magnetization relaxation. Single-crystal magnetization versus applied dc field scans exhibited hysteresis loops for both compounds, establishing them as new single-molecule and single-chain magnets, respectively. Complex 2 also showed steps in its hysteresis loops characteristic of quantum tunneling of magnetization, the highest nuclearity molecule to show such QTM steps. Arrhenius plots constructed from dc magnetization versus time decay plots gave effective barriers to magnetization relaxation (U(eff)) of 6 and 11 cm(-1) for 2 and 3, respectively. PMID- 17910438 TI - [Os(bipy)(CN)4]2- and its relatives as components of polynuclear assemblies: structural and photophysical properties. AB - A series of diimine-tetracyanoosmate anions [Os(diimine)(CN)4]2- [diimine=2,2' bipyridine (bipy), 2,2'-bipyrimidine (bpym), 1,10-phenanthroline (phen), and 4,4' tBu2-2,2'-bipyridine (tBu2bpy)] were prepared and isolated as their Na+ salts (water soluble) or PPN+ salts (soluble in organic solvents). Several examples were crystallographically characterized; the Na+ salts form a range of 1D, 2D, or 3D infinite coordination polymers via coordination of the cyanide groups to Na+ cations in either an end-on or a side-on manner. The [Os(diimine)(CN)4]2- anions are solvatochromic, showing three MLCT absorptions, which are considerably blue shifted in water compared to organic solvents, in the same way as is well-known for the analogous [Ru(diimine)(CN)4]2- anions. Luminescence in the red region of the spectrum is very weak but (following the expected solvatochromic behavior) is higher energy and more intense in water. However, by exploiting the effect of metallochromism (ref 4), the emission from [Os(tBu2bpy)(CN)4]2- in MeCN can be very substantially boosted in energy, intensity, and lifetime in the presence of Lewis-acidic metal cations (Na+, Ba2+, Zn2+), which, in a relatively noncompetitive solvent, coordinate to the cyanide groups of [Os(tBu2bpy)(CN)4]2-. This has an effect similar in principle to hydrogen bonding of the cyanides to delta+ protons of water, but very much stronger, such that in the presence of Zn2+ ions in MeCN the 1MLCT and 3MLCT absorptions are blue-shifted by ca. 7000 cm(-1), and the luminescence moves from 970 nm (vanishingly weak) to 610 nm with a lifetime of 120 ns (dominant component). Thus, the binding of metal cations to the cyanides provides a mechanism to incorporate [Os(diimine)(CN)4]2- complexes into polynuclear assemblies and simultaneously increases their 3MLCT energy and lifetime to an extent that makes them comparable to much-stronger luminophores such as Ru(II)-polypyridines. PMID- 17910439 TI - Synthesis, structure, spectral and electrochemical properties, and catalytic use of cobalt(III)-oxo cubane clusters. AB - Olive-green cobalt(III) complexes having the general formula Co4O4(O2CMe)4L4 (1) where L = py (1a), 4-Mepy (1b), 4-Etpy (1c), and 4-CNpy (1d) have been prepared by the H2O2 oxidation of a mixture of Co2+, MeCO2-, and pyridine or 4-substituted pyridines in a 1:2:1 molar ratio in methanol. Spectroscopic and X-ray crystallographic studies show that these complexes contain a tetrameric cubane like core [Co4(mu3-O)4]4+ where the four cobalt atoms form an approximate tetrahedron with edge lengths of approximately 2.75 A. Each cobalt in the crystallographically determined structure of Co4(mu3-O)4(mu-O2CMe)4L4 in 1a.NaClO4.3.5H2O and 1b.3H2O is hexacoordinate. Infrared spectra of the complexes show characteristic bands near 700, 635, and 580 cm-1 due to the central cubane like core. 1H NMR spectra of the complexes show that the dissolved species are essentially diamagnetic and also that the complexes maintain their integrity in solution. UV-vis spectra of the green solutions have been interpreted in terms of ligand-field and charge-transfer bands. The electrochemical behavior of the complexes studied by cyclic and differential pulse voltammetric techniques indicates that the [(CoIII)4(mu3-O)4]4+ core present in the complexes undergoes a reversible one-electron oxidation to the [(CoIII)3CoIV(mu3-O)4]5+ core with an E1/2 value below 1 V. This suggests that these complexes of cobalt may be suitable as catalysts for the oxidation of organic compounds. Preliminary investigations indicate that 1a has a role to play in the cobalt-catalyzed aerobic oxidation of neat ethylbenzene and p-xylene. PMID- 17910440 TI - Phosphine-substituted dithiolene complexes as ligands: communication between ruthenium(II) centers through a dimolybdenum bis(dithiolene) core. AB - The reaction of Mo2(SCH2CH2S)2Cp2 (1; Cp=eta-C5H5) with an excess of an alkyne in refluxing dichloromethane affords the bis(dithiolene) complexes Mo2(micro SCR1=CR2S)2Cp2 (2a, R1=R2=CO2Me; 2b, R1=R2=Ph; 2c, R1=H, R2=CO2Me) whereas with 1 equiv of alkyne at room temperature the mixed dithiolene-dithiolate species Mo2(micro-SCR1=CR2S)(micro-SCH2CH2S)Cp2 (3a, R1=R2=CO2Me; 3b, R1=R2=Ph) are formed. The remaining dithiolate ligand in 3 can then be converted into a different dithiolene by reaction with a second alkyne. Applying this methodology, we have used bis(diphenylphosphino)acetylene to prepare the first examples of complexes containing phosphine-substituted dithiolene ligands: Mo2{micro SC(CO2Me)=C(CO2Me)S}{micro-SC(PPh2)=C(PPh2)S}Cp2 (2g) and Mo2{micro SC(PPh2)=C(PPh2)S}2Cp2 (2h). Tri- and tetrametallic complexes can then be assembled by coordination of these diphosphines to CpRuCl units by reaction with CpRu(PPh3)2Cl. Electrochemical studies of the Ru(II)/Ru(III) couple in Mo2{micro SC(PPh2)=C(PPh2)S}2Cp2(RuClCp)2 (4b) reveals that the two separate ruthenium centers are oxidized electrochemically at different potentials, demonstrating communication between them through the dimolybdenum bis(dithiolene) core. Density functional theory calculations were carried out to explore the electronic structures of these species and to predict and assign their electronic spectra. PMID- 17910441 TI - Structural varieties in heterobimetallic lanthanide disiloxanediolates: "inorganic metallocenes" versus in-plane metallacrowns. AB - The previously proposed concept of "inorganic metallocenes" of group 3 and rare earth elements has been tested by preparing a series of novel disiloxanediolates with metals displaying different ionic radii. For the smaller scandium and yttrium, approximately planar arrangements of the disiloxanediolate frameworks with solvent and chloride ligands in trans positions were found. Thus, the compounds [{(Ph2SiO)2O}2{Li(DME)}2]ScCl(THF/DME) (2; DME=1,2-dimethoxyethane and THF=tetrahydrofuran) and [{(Ph2SiO)2O}2{Li(THF)2}2]YCl(THF) (3) can be described as heterobimetallic inorganic ring systems or metallacrown complexes with "in plane" coordination of the metal. In contrast, "out-of-plane" geometries with cis coordination of additional ligands were identified in the praseodymium derivatives [{(Ph2SiO)2O}2{Li(THF)2}{Li(THF)}]Pr(micro-Cl)2Li(THF)2 (4) and [{(Ph2SiO)2O}2{Li(DME)}2]PrCl(DME) (5). These compounds can be viewed as analogues of the known metallocene derivatives (C5Me5)2Pr(micro-Cl)2Li(THF)2 and (C5Me5)2PrCl(THF). The molecular structures of 2-5 have been determined by X-ray diffraction. PMID- 17910442 TI - Charge separation in a ruthenium-quencher conjugate bound to DNA. AB - A novel tris heteroleptic dipyridophenazine complex of ruthenium(II), [{Ru(phen)(dppz)(bpy'-his)}{Ru(NH3)5}]5+, containing a covalently tethered ruthenium pentammine quencher coordinated through a bridging histidine has been synthesized and characterized spectroscopically and biochemically in a DNA environment and in organic solvent. Steady-state and time-resolved luminescence measurements indicate that the tethered Ru complex is quenched relative to the parent complexes [Ru(phen)(dppz)(bpy')]2+ and [Ru(phen)(dppz)(bpy'-his)]2+ in DNA and acetonitrile, consistent with intramolecular photoinduced electron transfer. Intercalated into guanine-containing DNA, [{Ru(phen)(dppz)(bpy' his)}{Ru(NH3)5}]5+, upon excitation and intramolecular quenching, is capable of injecting charge into the duplex based upon the EPR detection of guanine radicals. DNA-mediated charge transport is also indicated using a kinetically fast cyclopropylamine-substituted base as an electron hole trap. Guanine damage is not observed, however, in measurements using the guanine radical as the kinetically slower hole trap, indicating that back electron-transfer reactions are competitive with guanine oxidation. Moreover, transient absorption measurements reveal a novel photophysical reaction pathway for [{Ru(phen)(dppz)(bpy'-his)}{Ru(NH3)5}]5+ in the presence of DNA that is competitive with the intramolecular flash-quench process. These results illustrate the remarkably rich redox chemistry that can occur within a bimolecular ruthenium complex intercalated in duplex DNA. PMID- 17910443 TI - Enhancement in electronic communication upon replacement of Mo-O by Mo-S bonds in tetranuclear clusters of the type [Mo2]2(mu-E-E)2 (E = O or S). AB - A tetranuclear cluster containing two quadruply bonded cis-Mo2(DAniF)2 units (DAniF = N,N'-di-p-anisylformamidinate) linked by four hydroxide groups (1) was obtained by hydrolysis of [Mo2(cis-DAniF)2](mu-OCH3)4. Analogous compounds linked by two bidentate bridges (o-O2C6H4 for 2, o-O2C10H6 for 3, and o-S2C6H4 for 4) were synthesized by direct assembly of the corner species precursor [Mo2(cis DAniF)2(NCCH3)4](BF4)2 and the respective protonated ligands. All four compounds were characterized by X-ray crystallography. Cyclic voltammograms of the O-linked compound 2 and the S analogue 4 show two reversible one-electron-oxidation processes with potential separations (DeltaE(1/2)) of 474 and 776 mV, respectively. The large increase of about 300 mV in DeltaE(1/2) for the S analogue relative to that of the O compound is consistent with a large increase in electronic communication. This enhancement occurs despite the increase of ca. 0.45 A in nonbonding separation between the midpoints of the Mo2 units, which changes from 3.266 A in 2 to 3.72 A in 4, and the increase of ca. 0.4 A in M-E distances as E changes from O to S. Density functional theory calculations show that the increase in electronic communication between the metal centers in 4 is due to a superexchange pathway involving d and p orbitals in the linker E atoms that is less important in 2. PMID- 17910444 TI - Diverse stereocontrol effects induced by weakly coordinating anions. Stereospecific olefin polymerization pathways at archetypal C(s)- and C(1) symmetric metallocenium catalysts using mono- and polynuclear halo perfluoroarylmetalates as cocatalysts. AB - Counteranion effects on propylene polymerization rates and stereoselectivities are compared using Cs-symmetric Me2C(Cp)(Flu)ZrMe2 (1; Cp = C5H4,eta5 cyclopentadienyl; Flu = C13H8, eta5-fluorenyl) and C1-symmetric Me2Si(OHF)(CpR*)ZrMe2 (2; OHF = C13H16, eta5-octahydrofluorenyl; CpR* = eta5-3-( )-menthylcyclopentadienyl) precatalysts activated with the mononuclear and polynuclear perfluoroarylborate, -aluminate, and -gallate cocatalysts/activators B(C6F5)3 (3), B(o-C6F5C6F4)3 (4), Al(C6F5)3 (5), Ph3C+B(C6F5)4- (6) Ph3C+FAl(o C6F5C6F4)3- (7), Ga(C6F5)3 (8), and recently reported mono- and polymetallic trityl perfluoroarylhalometalates Ph3C+FB(C6F5)3- (9), Ph3C+FB(o-C6F5C6F4)3- (10), (Ph3C+)xFx[Al(C6F5)3]yx- (x = 1, y = 1, 11; x = 1, y = 2, 12; x = 2, y = 3, 13), Ph3C+(C6F5)3AlFAl(o-C6F5C6F4)3- (14), Ph3C+XAl(C6F5)3- (X = Cl, 15; X = Br, 16), and Ph3C+F[Ga(C6F5)3]2- (17). Temperature, propylene concentration, and solvent polarity dependence are surveyed in polymerizations catalyzed by 1 activated with cocatalysts 3-16 and with a 1:2 ratio of Ph3CCl and 5, and with a 1:2 ratio of Ph3CBr and 5, and by 2 activated with 3, 6, 7, 12, and 14. Remarkable stereocontrol with high activities is observed for 1 + 12 and 1 + 14. Polypropylene samples produced using C1-symmetric precatalyst 2 are subjected to microstructural analyses using stochastic models describing the relative contributions of enantiofacial misinsertion and backskip processes. A powerful technique is introduced for calculating interparametric correlation matrices for these nonlinear stochastic models. The collected results significantly extend what is known about ion-pairing effects in the case of Cs-symmetric precatalyst 1 and allow these findings to be applied to the case of C1-symmetric precatalyst 2 as an agent of isospecific propylene polymerization. PMID- 17910445 TI - On the kinetic and thermodynamic reactivity of lithium di(alkyl)amidozincate bases in directed ortho metalation. AB - Sequential reaction of HTMP (= 2,2,6,6-tetramethylpiperidine) with nBuLi and Et2Zn affords unsolvated polymer chains of EtZn(micro-Et)(micro-TMP)Li 6. The scope of this reagent in directed ortho metalation (DoM) chemistry has been tested by its reaction with N,N-diisopropylnaphthamide in THF to give EtZn(micro C10H6C(O)NiPr2-2)2Li.2THF 7. Data reveal that 6 has undergone reaction with 2 equiv of aromatic tertiary amide and imply that it exhibits dual alkyl/amido basicity. DFT calculations reveal that direct alkyl basicity is kinetically disfavored and instead point to a stepwise mechanism whereby 6 acts as an amido base, liberating HTMP during the first DoM event. Re-coordination of the amine at lithium then incurs the elimination of EtH. Reaction of the resulting alkyl(amido)(arylamido)zincate with a second equivalent of N,N diisopropylnaphthamide eliminates HTMP and affords 7. Both DoM steps involve the exhibition of amido basicity and each reveals a low kinetic barrier to reaction. Understanding of this reaction sequence is tested by treating 6 with N,N diisopropylbenzamide in THF. On the basis of theory and experiment, the presence of THF solvent (in place of stronger Lewis bases) combined with the use of a sterically less congested aromatic amide is expected to encourage threefold, stepwise reaction. Isolation and characterization of the resulting tripodal zincate Zn(micro-C6H4C(O)NiPr2-2)3Li.THF 8 bears this out and suggests a significant new level of control in zincate-induced DoM chemistry through the combination of experiment and DFT studies. PMID- 17910446 TI - Conformational dependence of hemoglobin reactivity under high viscosity conditions: the role of solvent slaved dynamics. AB - The concept of protein dynamic states is introduced. This concept is based on (i) protein dynamics being organized hierarchically with respect to solvent slaving and (ii) which tier of dynamics is operative over the time window of a given measurement. The protein dynamic state concept is used to analyze the kinetic phases derived from the recombination of carbon monoxide to sol-gel-encapsulated human adult hemoglobin (HbA) and select recombinant mutants. The temperature dependent measurements are made under very high viscosity conditions obtained by bathing the samples in an excess of glycerol. The results are consistent with a given tier of solvent slaved dynamics becoming operative at a time delay (with respect to the onset of the measurement) that is primarily solvent- and temperature-dependent. However, the functional consequences of the dynamics are protein- and conformation-specific. The kinetic traces from both equilibrium populations and trapped allosteric intermediates show a consistent progression that exposes the role of both conformation and hydration in the control of reactivity. Iron-zinc symmetric hybrid forms of HbA are used to show the dramatic difference between the kinetic patterns for T state alpha and beta subunits. The overall results support a model for allostery in HbA in which the ligand-binding induced transition from the deoxy T state to the high -affinity R state proceeds through a progression of T state intermediates. PMID- 17910447 TI - Explanation of the unusual temperature dependence of the atmospherically important OH + H(2)S --> H(2)O + HS reaction and prediction of the rate constant at combustion temperatures. AB - Rate constants for the OH + H2S --> H2O + HS reaction, which is important for both atmospheric chemistry and combustion, are calculated by direct dynamics with the M06-2X density functional using the MG3S basis set. Energetics are compared to high-level MCG3/3//MC-QCISD/3 wave function theory and to results obtained by other density functionals. We employ canonical variational transition-state theory with multidimensional tunneling contributions and scaled generalized normal-mode frequencies evaluated in redundant curvilinear coordinates with anharmonicity included in the torsion. The transition state has a quantum mechanically distinguishable, nonsuperimposable mirror image that corresponds to a separate classical reaction path; the effect of the multiple paths is examined through use of a symmetry number and by torsional methods. Calculations with the reference-potential Pitzer-Gwinn treatment of the torsional mode agree with experiment, within experimental scatter, and predict a striking temperature dependence of the activation energy, increasing from -0.1 kcal/mol at 200 K to 0.2, 1.0, 3.4, and 9.8 kcal/mol at 300, 500, 1000, and 2400 K. The unusual temperature dependence arises from a dynamical bottleneck at an energy below reactants, following an addition complex on the reaction path with a classical binding energy of 4.4 kcal/mol. As a way to check the mechanism, kinetic isotope effects of the OH + D2S and OD + D2S reactions have been predicted. PMID- 17910448 TI - Paramagnetism-based NMR restraints provide maximum allowed probabilities for the different conformations of partially independent protein domains. AB - An innovative analytical/computational approach is presented to provide maximum allowed probabilities (MAPs) of conformations in protein domains not rigidly connected. The approach is applied to calmodulin and to its adduct with alpha synuclein. Calmodulin is a protein constituted by two rigid domains, each of them composed by two calcium-binding EF-hand motifs, which in solution are largely free to move with respect to one another. We used the N60D mutant of calmodulin, which had been engineered to selectively bind a paramagnetic lanthanide ion to only one of its four calcium binding sites, specifically in the second EF-hand motif of the N-terminal domain. In this way, pseudocontact shifts (pcs's) and self-orientation residual dipolar couplings (rdc's) measured on the C-terminal domain provide information on its relative mobility with respect to the domain hosting the paramagnetic center. Available NMR data for terbium(III) and thulium(III) calmodulin were supplemented with additional data for dysprosium(III), analogous data were generated for the alpha-synuclein adduct, and the conformations with the largest MAPs were obtained for both systems. The MAP analysis for calmodulin provides further information on the variety of conformations experienced by the system. Such variety is somewhat reduced in the calmodulin-alpha-synuclein adduct, which however still retains high flexibility. The flexibility of the calmodulin-alpha-synuclein adduct is an unexpected result of this research. PMID- 17910449 TI - Enthalpic and entropic stages in alpha-helical peptide unfolding, from laser T jump/UV Raman spectroscopy. AB - The alpha-helix is a ubiquitous structural element in proteins, and a number of studies have addressed the mechanism of helix formation and melting in simple peptides. However, fundamental issues remain to be resolved, particularly the temperature (T) dependence of the rate. In this work, we report application of a novel kHz repetition rate solid-state tunable NIR (pump) and deep UV Raman (probe) laser system to study the dynamics of helix unfolding in Ac-GSPEA3KA4KA4 CO-D-Arg-CONH2, a peptide designed for helix stabilization in aqueous solution. Its T-dependent UV resonance Raman (UVRR) spectra, excited at 197 nm for optimal enhancement of amide vibrations, were decomposed into variable contributions from helix and coil spectra. The helix fractions derived from the UVRR spectra and from far UV CD spectra were coincident at low T but deviated increasingly at high T, the UVRR curve giving higher helix content. This difference is consistent with the greater sensitivity of UVRR spectra to local conformation than CD. After a laser-induced T-jump, the UVRR-determined helix fractions defined monoexponential decays, with time-constants of approximately 120 ns, independent of the final T (Tf = 18-61 degrees C), provided the initial T (Ti) was held constant (6 degrees C). However, there was also a prompt loss of helicity, whose amplitude increased with increasing Tf, thereby defining an initial enthalpic phase, distinct from the subsequent entropic phase. These phases are attributed to disruption of H bonds followed by reorientation of peptide links, as the chain is extended. When Ti was raised in parallel with Tf (10 degrees C T-jumps), the prompt phase merged into an accelerating slow phase, an effect attributable to the shifting distribution of initial helix lengths. Even greater acceleration with rising Ti has been reported in T-jump experiments monitored by IR and fluorescence spectroscopies. This difference is attributable to the longer range character of these probes, whose responses are therefore more strongly weighted toward the H bond-breaking enthalpic process. PMID- 17910450 TI - Preparation and controlled self-assembly of Janus magnetic nanoparticles. AB - Janus magnetic nanoparticles (~20 nm) were prepared by grafting either polystyrene sodium sulfonate (PSSNa) or polydimethylamino ethylmethacrylate (PDMAEMA) to the exposed surfaces of negatively charged poly(acrylic acid) (PAA) coated magnetite nanoparticles adsorbed onto positively charged silica beads. Individually dispersed Janus nanoparticles were obtained by repulsion from the beads on reversal of the silica surface charge when the solution pH was increased. Controlled aggregation of the Janus nanoparticles was observed at low pH values, with the formation of stable clusters of approximately 2-4 times the initial size of the particles. Cluster formation was reversed, and individually dispersed nanoparticles recovered, by restoring the pH to high values. At intermediate pH values, PSSNa Janus nanoparticles showed moderate clustering, while PDMAEMA Janus nanoparticles aggregated uncontrollably due to dipolar interactions. The size of the stable clusters could be controlled by increasing the molecular weight of the grafted polymer, or by decreasing the magnetic nanoparticle surface availability for grafting, both of which yielded larger cluster sizes. The addition of small amounts of PAA-coated magnetic nanoparticles to the Janus nanoparticle suspension resulted in a further increase in the final cluster size. Monte Carlo simulation results compared favorably with experimental observations and showed the formation of small, elongated clusters similar in structure to those observed in cryo-TEM images. PMID- 17910451 TI - Detection of an eta(1)-alkene intermediate of the type [Cp2Zr(Me)(eta(1) alkene)]+: the role of such species in metallocene catalyst deactivation to allylic species. PMID- 17910452 TI - Electrocatalysis on bimetallic surfaces: modifying catalytic reactivity for oxygen reduction by voltammetric surface dealloying. PMID- 17910453 TI - Voltammetry of quinones in unbuffered aqueous solution: reassessing the roles of proton transfer and hydrogen bonding in the aqueous electrochemistry of quinones. AB - Cyclic voltammetry studies are reported for two representative quinones, benzoquinone and 2-anthraquinonesulfonate, in buffered and unbuffered aqueous solution at different pH's. While the redox reaction of quinones in buffered water is well described as an overall 2 e-, 2 H+ reduction to make the hydroquinone, a much better description of the overall reaction in unbuffered water is as a 2 e- reduction to make the strongly hydrogen-bonded quinone dianion, which will exist in water as an equilibrium mixture of protonation states. This description helps to unify quinone electrochemistry by bridging the apparent gap between the redox chemistry of quinones in water and that in aprotic organic solvents, where quinones undergo two sequential 1 e- reductions to form the quinone dianion. PMID- 17910455 TI - Stable analogues of aminoacyl-tRNA for inhibition of an essential step of bacterial cell-wall synthesis. PMID- 17910454 TI - Dual-mode fluorophore-doped nickel nitrilotriacetic acid-modified silica nanoparticles combine histidine-tagged protein purification with site-specific fluorophore labeling. AB - We present the first example of a fluorophore-doped nickel chelate surface modified silica nanoparticle that functions in a dual mode, combining histidine tagged protein purification with site-specific fluorophore labeling. Tetramethylrhodamine (TMR)-doped silica nanoparticles, estimated to contain 700 900 TMRs per ca. 23 nm particle, were surface modified with nitrilotriacetic acid (NTA), producing TMR-SiO2-NTA-Ni2+. Silica-embedded TMR retains very high quantum yield, is resistant to quenching by buffer components, and is modestly quenched and only to a certain depth (ca. 2 nm) by surface-attached Ni2+. When exposed to a bacterial lysate containing estrogen receptor alpha ligand binding domain (ERalpha) as a minor component, these beads showed very high specificity binding, enabling protein purification in one step. The capacity and specificity of these beads for binding a his-tagged protein were characterized by electrophoresis, radiometric counting, and MALDI-TOF MS. ERalpha, bound to TMR-SiO2-NTA-Ni++ beads in a site-specific manner, exhibited good activity for ligand binding and for ligand-induced binding to coactivators in solution FRET experiments and protein microarray fluorometric and FRET assays. This dual-mode type TMR-SiO2-NTA-Ni2+ system represents a powerful combination of one-step histidine-tagged protein purification and site-specific labeling with multiple fluorophore species. PMID- 17910457 TI - Programmed thermodynamic formation and structure analysis of star-like nanogels with core cross-linked by thermally exchangeable dynamic covalent bonds. AB - Programmed thermodynamic formation of star-like nanogels from designed diblock copolymers with thermally exchangeable dynamic covalent bonds in their side chains and structure analysis of the nanogels were performed. Linear diblock copolymers that consist of poly(methyl methacrylate) block and random copolymer block of methyl methacrylate (MMA) and methacrylic esters with alkoxyamine moiety were prepared by atom transfer radical polymerization (ATRP). By heating the diblock copolymers in anisole, a cross-linking reaction occurred as a result of the radical crossover reaction of alkoxyamine moieties to afford star-like nanogels. Kinetic studies have revealed that the cross-linking behavior reaches equilibrium at a given reaction time, with characteristic reaction behaviors for thermodynamic reactions being observed. The equilibrium structures of the star like nanogels were controlled by the initial concentrations of diblock copolymers as well as their compositions and molecular weights. Furthermore, by heating the star-like nanogels with excess alkoxyamine, linear polymers were successfully regenerated. The molecular weights and sizes of the nanogels were evaluated by gel permeation chromatography-multiangle laser light scattering (GPC-MALLS) and small-angle X-ray scattering (SAXS) measurements, respectively, and the morphologies of the nanogels were directly observed by scanning force microscopy (SFM). PMID- 17910456 TI - Zirconocene-mediated highly regio- and stereoselective synthesis of multisubstituted olefins starting from 1-alkynylboronates. PMID- 17910458 TI - How small variations in crystal interactions affect macroscopic properties. PMID- 17910460 TI - Effective modulation of the donor properties of N-heterocyclic carbene ligands by "through-space" communication within a planar chiral scaffold. PMID- 17910459 TI - Probing supramolecular structure from measurement of methyl (1)H-(13)C residual dipolar couplings. PMID- 17910462 TI - Studies on the generation of unnatural C-nucleosides with 1-alkynyl-2-deoxy-D riboses. AB - 1-alkynyl-2-deoxy-D-riboses 7 and 8 were independently synthesized and subsequently used to generate several novel C-nucleosides. PMID- 17910461 TI - A stereoselective intramolecular halo-etherification of chiral enamides in the synthesis of halogenated cyclic ethers. AB - A stereoselective halo-etherification of chiral enamides is described here. This work provides an approach to halogen containing cyclic ethers and reveals further mechanistic insights to the chemistry of chiral enamides. PMID- 17910463 TI - Conformational equilibria of ethanolamine and its hydrochloride in solution. AB - The conformational preferences of ethanolamine and its hydrochloride in solution were estimated by comparing experimental NMR vicinal proton-proton coupling constants to semiemprical coupling constants for each staggered rotamer, derived by the Haasnoot-Altona method. Strong gauche preferences are observed for both ethanolamine and its hydrochloride over a wide range of solvent polarities. Concentration was not observed to significantly affect the position of the conformer equilibria. PMID- 17910465 TI - Intermolecular cross-double-michael addition between nitro and carbonyl activated olefins as a new approach in C-C bond formation. AB - A novel intermolecular cross-double-Michael addition between nitro and carbonyl activated olefins has been developed through Lewis base catalysis. The reaction took place with a large group of beta-alkyl nitroalkenes and alpha,beta unsaturated ketone/esters, producing an allylic nitro compound in good to excellent yields. PMID- 17910464 TI - Selectively chemodosimetric detection of Hg(II) in aqueous media. AB - A new series of benzoylthiourea derivatives of 1, 3, and 5 were prepared, and their chemodosimetric behaviors toward metal cations were investigated in aqueous media at room temperature. Among various metal cations tested, exclusively Hg2+ ion responses to irreversible color changes of receptors, along with distinctive blue shifts in UV/vis spectra. The receptors can be applicable for the monitoring of Hg2+ ion in aqueous solution with a pH span 4-9. PMID- 17910466 TI - Preparation of alpha-haloacrylate derivatives via dimethyl sulfoxide-mediated selective dehydrohalogenation. AB - Dimethyl sulfoxide causes alpha,beta-dihalopropanoate derivatives to undergo efficient, selective dehydrohalogenation to form alpha-haloacrylate analogues. A variety of alpha-halo Michael acceptors were prepared in dimethyl sulfoxide under mild, base-free conditions, including the preparation of alpha-bromoacrolein and alpha-chloro- and bromoacrylonitriles. Synthesis of these molecules has been reported in the literature to be difficult. Among all the existing dehydrohalogenation procedures, this protocol is the most facile, practical, and environmentally benign process. PMID- 17910467 TI - Anodic coupling reactions: a sequential cyclization route to the arteannuin ring skeleton. AB - A pair of intramolecular anodic olefin coupling reactions has been used to construct the arteannuin ring skeleton. Both coupling reactions took advantage of a furan ring as one of the coupling partners. In the first, it was found that an enol ether derived from an aldehyde was not an effective initiating group for the reaction. Instead, the cyclization benefited strongly from the use of a N,O ketene acetal initiating group. In the second cyclization, an endocyclic enol ether was coupled to the furan ring. This second electrolysis reaction generated the key tetrasubstituted carbon at the center of the arteannuin ring skeleton. PMID- 17910468 TI - A new, powerful glycosylation method: activation of thioglycosides with dimethyl disulfide-triflic anhydride. AB - Dimethyl disulfide reacts with triflic anhydride to provide a highly reactive electrophile. Various thioglycosides, differing in their thio aglycons, carbohydrate units, and protecting group pattern, were activated with Me2S2-Tf2O in the presence of different glycosyl acceptors. The reactions proceeded at low temperatures within a short time, affording oligosaccharides in high yields both on primary and secondary hydroxyls. Armed and disarmed glycosyl donors were activated equally efficiently. PMID- 17910469 TI - Synthesis of tertiary alpha-hydroxy acids by silylene transfer to alpha-keto esters. AB - Alpha-keto esters can be converted into alpha-hydroxy acids in a single flask involving metal-catalyzed silylene transfer, 6pi-electrocyclization, Ireland Claisen rearrangement, and hydrolysis. This reaction sequence is stereoselective and tolerates alkyl- and aryl-substituted alpha-keto ester substrates as well as an alpha-imino ester. PMID- 17910471 TI - Sequence recognition in the minor groove of DNA by covalently linked formamido imidazole-pyrrole-imidazole polyamides: effect of H-pin linkage and linker length on selectivity and affinity. AB - The polyamide N-formamido imidazole-pyrrole-imidazole (f-ImPyIm) binds with an exceptionally high affinity for its cognate site 5'-ACGCGT-3' as a stacked, staggered, and noncovalent cooperative dimer. Investigations are presented into its sequence specificity and binding affinity when linked covalently as an H-pin "dimer". Five f-ImPyIm cross-linked analogues with six to nine methylene linkers and an eight-linked ethylene glycol linker were examined to investigate the effect of linkage and linker length on DNA binding. Thermal denaturation studies on short DNA hairpins showed preferential binding by both f-ImPyIm (DeltaTm = 7.8 degrees C) and its cross-linked derivatives (DeltaTm > 30 degrees C) at 5'-ACGCGT 3', indicating sequence specificity was retained on linkage. DNase I footprinting confirmed strict cognate site selectivity and demonstrated that affinity increased with linker length (f-ImPyIm-9 = f-ImPyIm-8 = f-ImPyIm-EG-8 > f-ImPyIm 7 > f-ImPyIm-6). The eight- and nine-linked derivatives bound at 100-fold lower concentrations at the cognate site relative to f-ImPyIm-6, and with 10-fold higher affinity than unlinked f-ImPyIm. Use of an ethylene glycol linkage in f ImPyIm-EG-8 to improve solubility slightly increased the cognate site affinity relative to those of f-ImPyIm-8 and f-ImPyIm-9, although some selectivity was lost at high ligand concentration. CD demonstrated that cognate site binding by eight and nine-linked compounds occurred in the minor groove. SPR analysis gave a binding affinity (K) for f-ImPyIm-EG-8 at the cognate site of 2 x 10(10) M-1, representing a 100-fold increase relative to that of f-ImPyIm. This study demonstrates that the high-affinity cooperative binding of f-ImPyIm can be enhanced significantly by suitable covalent linkage, while maintaining its strict cognate site selectivity. PMID- 17910470 TI - Calcium regulation of calmodulin binding to and dissociation from the myo1c regulatory domain. AB - Myo1c is an unconventional myosin involved in cell signaling and membrane dynamics. Calcium binding to the regulatory-domain-associated calmodulin affects myo1c motor properties, but the kinetic details of this regulation are not fully understood. We performed actin gliding assays, ATPase measurements, fluorescence spectroscopy, and stopped-flow kinetics to determine the biochemical parameters that define the calmodulin-regulatory-domain interaction. We found calcium moderately increases the actin-activated ATPase activity and completely inhibits actin gliding. Addition of exogenous calmodulin in the presence of calcium fully restores the actin gliding rate. A fluorescently labeled calmodulin mutant (N111C) binds to recombinant peptides containing the myo1c IQ motifs at a diffusion-limited rate in the presence and absence of calcium. Measurements of calmodulin dissociation from the IQ motifs in the absence of calcium show that the calmodulin bound to the IQ motif adjacent to the motor domain (IQ1) has the slowest dissociation rate (0.0007 s-1), and the IQ motif adjacent to the tail domain (IQ3) has the fastest dissociation rate (0.5 s-1). When the complex is equilibrated with calcium, calmodulin dissociates most rapidly from IQ1 (60 s-1). However, this increased rate of dissociation is limited by a slow calcium-induced conformational change (3 s-1). Fluorescence anisotropy decay of fluorescently labeled N111C bound to myo1c did not depend appreciably on Ca2+. Our data suggest that the calmodulin bound to the IQ motif adjacent to the motor domain is rapidly exchangeable in the presence of calcium and is responsible for regulation of myo1c ATPase and motile activity. PMID- 17910472 TI - Two distinct proton binding sites in the ATP synthase family. AB - The F1F0 ATP synthase utilizes energy stored in an electrochemical gradient of protons (or Na+ ions) across the membrane to synthesize ATP from ADP and phosphate. Current models predict that the protonation/deprotonation of specific acidic c ring residues is at the core of the proton translocation mechanism by this enzyme. To probe the mode of proton binding, we measured the covalent modification of the acidic c ring residues with the inhibitor dicyclohexylcarbodiimide (DCCD) over the pH range from 5 to 11. With the H+ translocating ATP synthase from the archaeum Halobacterium salinarium or the Na+ translocating ATP synthase from Ilyobacter tartaricus, the pH profile of DCCD labeling followed a titration curve with a pKa around neutral, reflecting protonation of the acidic c ring residues. However, with the ATP synthases from Escherichia coli, mitochondria, or chloroplasts, a clearly different, bell-shaped pH profile for DCCD labeling was observed which is not compatible with carboxylate protonation but might be explained by the coordination of a hydronium ion as proposed earlier [Boyer, P. D. (1988) Trends Biochem. Sci. 13, 5-7]. Upon site-directed mutagenesis of single binding site residues of the structurally resolved c ring, the sigmoidal pH profile for DCCD labeling could be converted to a more bell-shaped one, demonstrating that the different ion binding modes are based on subtle changes in the amino acid sequence of the protein. The concept of two different binding sites in the ATP synthase family is supported by the ATP hydrolysis pH profiles of the investigated enzymes. PMID- 17910473 TI - NMR solution structure of subunit F of the methanogenic A1AO adenosine triphosphate synthase and its interaction with the nucleotide-binding subunit B. AB - The A1AO adenosine triphosphate (ATP) synthase from archaea uses the ion gradients generated across the membrane sector (AO) to synthesize ATP in the A3B3 domain of the A1 sector. The energy coupling between the two active domains occurs via the so-called stalk part(s), to which the 12 kDa subunit F does belong. Here, we present the solution structure of the F subunit of the A1AO ATP synthase from Methanosarcina mazei Go1. Subunit F exhibits a distinct two-domain structure, with the N-terminal having 78 residues and residues 79-101 forming the flexible C-terminal part. The well-ordered N-terminal domain is composed of a four-stranded parallel beta-sheet structure and three alpha-helices placed alternately. The two domains are loosely associated with more flexibility relative to each other. The flexibility of the C-terminal domain is further confirmed by dynamics studies. In addition, the affinity of binding of mutant subunit F, with a substitution of Trp100 against Tyr and Ile at the very C terminal end, to the nucleotide-binding subunit B was determined quantitatively using the fluorescence signals of natural subunit B (Trp430). Finally, the arrangement of subunit F within the complex is presented. PMID- 17910474 TI - Characterization of Rat NTPDase1, -2, and -3 ectodomains refolded from bacterial inclusion bodies. AB - The ecto-nucleoside triphosphate diphosphohydrolases or NTPDases are a family of membrane-bound enzymes that catalyze the sequential removal of gamma- and beta phosphate from ATP, ADP, and other nucleotides. NTPDase1, -2, -3, and -8 are the enzymes responsible for signal conversion and termination in purinergic signaling. They are anchored to the cytoplasmic membrane by two transmembrane helices with a large catalytic domain pointing toward the extracellular space. Here we report the first successful expression and purification of the soluble extracellular domains of rat NTPDase1, -2, and -3 from bacterial inclusion bodies. The refolded proteins show characteristics similar to the wild type enzymes, for example in that they are dependent on divalent metal ions for catalysis and hydrolyze a wide variety of nucleoside tri- and diphosphates, whereas the monophosphate AMP is not further degraded. Nucleoside triphosphates are hydrolyzed at a higher rate than the corresponding diphosphates. Other characteristics of the recombinant enzymes however reflect the absence of transmembrane regions and side chain glycosylation. For example all three enzymes are monomeric and only subtly activated by Mg2+ ions as compared to Ca2+ ions. Although having a considerably higher specificity constant kcat/Km for ADP as for ATP, the bacterially expressed variant of NTPDase1 in contrast to its wild type counterpart releases intermediate ADP to a substantial amount. The presented expression system will allow large scale production of active protein suitable for structural studies, development of inhibitors, and even clinical application. PMID- 17910476 TI - Kinetics of cryptdin-4 translocation coupled with peptide-induced vesicle leakage. AB - The antimicrobial peptide cryptdin-4 (Crp4), a member of the alpha-defensin family, is shown to translocate cooperatively across phospholipid bilayers. The cooperativity of the process is manifested by translocation kinetics which vary with the peptide to lipid molar ratio. A simple association model suggests dimerization. Black lipid membrane experiments reveal that Crp4 translocation does not create well-defined aqueous pores, as is often common among peptides exhibiting cooperative translocation. Still, the efflux induced by Crp4 upon its interaction with fluorophore-loaded vesicles is shown to be a direct result of the membrane perturbation resulting from the translocation process. Leakage can be predicted by relating membrane permeability to the fraction of peptide translocated. Crp4 translocation has implications for its antimicrobial activity as internalized peptide would be available to attack intracellular targets. PMID- 17910477 TI - Amyloid-beta(1-42) rapidly forms protofibrils and oligomers by distinct pathways in low concentrations of sodium dodecylsulfate. AB - Alzheimer's disease (AD) is characterized by large numbers of senile plaques in the brain that consist of fibrillar aggregates of 40- and 42-residue amyloid-beta (Abeta) peptides. However, the degree of dementia in AD correlates better with the concentration of soluble Abeta species assayed biochemically than with histologically determined plaque counts, and several investigators now propose that soluble aggregates of Abeta are the neurotoxic agents that cause memory deficits and neuronal loss. These endogenous aggregates are minor components in brain extracts from AD patients and transgenic mice that express human Abeta, but several species have been detected by gel electrophoresis in sodium dodecylsulfate (SDS) and isolated by size exclusion chromatography (SEC). Endogenous Abeta aggregation is stimulated at cellular interfaces rich in lipid rafts, and anionic micelles that promote Abeta aggregation in vitro may be good models of these interfaces. We previously found that micelles formed in dilute SDS (2 mM) promote Abeta(1-40) fiber formation by supporting peptide interaction on the surface of a single micelle complex. In contrast, here we report that monomeric Abeta(1-42) undergoes an immediate conversion to a predominant beta structured conformation in 2 mM SDS which does not proceed to amyloid fibrils. The conformational change is instead rapidly followed by the near quantitative conversion of the 4 kDa monomer SDS gel band to 8-14 kDa bands consistent with dimers through tetramers. Removal of SDS by dialysis gave a shift in the predominant SDS gel bands to 30-60 kDa. While these oligomers resemble the endogenous aggregates, they are less stable. In particular, they do not elute as discrete species on SEC, and they are completed disaggregated by boiling in 1% SDS. It appears that endogenous oligomeric Abeta aggregates are stabilized by undefined processes that have not yet been incorporated into in vitro Abeta aggregation procedures. PMID- 17910478 TI - Pockets of short-range transient order and restricted topological heterogeneity in the guanidine-denatured state ensemble of GED of dynamin. AB - The nature and variety in the denatured state of a protein, a non-native state under a given set of conditions, has been a subject of intense debate. Here, using multidimensional NMR, we have characterized the 6 M Gdn-HCl-denatured state of GED, the assembly domain of dynamin. Even under such strongly denaturing conditions, we detected the presence of conformations in slow exchange on the NMR chemical shift time scale. Although the GED oligomer as well as the SDS-denatured monomeric GED were seen to be predominantly helical [Chugh et al. (2006) FEBS J. 273, 388-397], the 6 M Gdn-HCl-denatured GED has largely beta-structural preferences. However, against such a background, we could detect the presence of a population with a short helical stretch (Arg42-Ile47) in the ensemble. The 1H 1H NOEs suggested presence of pockets of transient short-range order along the chain. Put together these segments may lead to a rather small number of interconverting topologically distinguishable ensembles. Spectral density analysis of 15N relaxation rates and {1H}-15N NOE, measured at 600 and 800 MHz, and comparison of J(0) with hydrophobic patches calculated using AABUF approach, indicated presence of four domains of slow motions. These coincided to a large extent with those showing significant Rex. Additionally, a proline residue in the connection between two of these domains seems to cause a fast hinge motion. These observations help enhance our understanding of protein denatured states, and of folding concepts, in general. PMID- 17910475 TI - Tyrosine nitration of IkappaBalpha: a novel mechanism for NF-kappaB activation. AB - The NF-kappaB family of transcription factors is an important component of stress activated cytoprotective signal transduction pathways. Previous studies demonstrated that some activation mechanisms require phosphorylation, ubiquitination, and degradation of the inhibitor protein, IkappaBalpha. Herein, it is demonstrated that ionizing radiation in the therapeutic dose range stimulates NF-kappaB activity by a mechanism in which IkappaBalpha tyrosine 181 is nitrated as a consequence of constitutive NO* synthase activation, leading to dissociation of intact IkappaBalpha from NF-kappaB. This mechanism does not appear to require IkappaBalpha kinase-dependent phosphorylation or proteolytic degradation of IkappaBalpha. Tyrosine 181 is involved in several noncovalent interactions with the p50 subunit of NF-kappaB stabilizing the IkappaBalpha-NF kappaB complex. Evaluation of hydropathic interactions of the IkappaBalpha-p50 complex on the basis of the crystal structure of the complex is consistent with nitration disrupting these interactions and dissociating the IkappaBalpha-NF kappaB complex. Tyrosine nitration is not commonly studied in the context of signal transduction. However, these results indicate that tyrosine nitration is an important post-translational regulatory modification for NF-kappaB activation and possibly for other signaling molecules modulated by mild and transient oxidative and nitrosative stresses. PMID- 17910479 TI - Photoactivated 3-azioctanol irreversibly desensitizes muscle nicotinic ACh receptors via interactions at alphaE262. AB - 3-Azioctanol is a photoactivatable analogue of octanol that noncompetitively inhibits nicotinic acetylcholine receptors (nAChRs). Photolabeling studies using [3H]-3-azioctanol in Torpedo nAChR identified alphaE262 as a site of desensitization-dependent incorporation. However, it is unknown whether photolabeling of alphaE262 causes functional effects in nAChRs and what other roles this residue plays in gating, desensitization, and channel block. We used ultrafast patch-perfusion electrophysiology and ultraviolet (UV) irradiation to investigate the state-dependence of both reversible nAChR inhibition by 3 azioctanol and the irreversible effects of photoactivated 3-azioctanol. Channels with mutations at alphaE262 were studied to determine ACh EC50s, desensitization rates, and sensitivities to reversible and photoirreversible 3-azioctanol inhibition. Exposure to 3-azioctanol in the presence of 365 nm UV light produced irreversible inhibition of wild-type nAChRs. Desensitization with ACh dramatically increased the degree of irreversible inhibition by photoactivated 3 azioctanol. Mutations at alphaE262 that reduce diazirine photomodification decreased the irreversible inhibition induced by photoactivated 3-azioctanol. Hydrophobic mutations at alphaE262 significantly slowed rapid ACh-induced desensitization and dramatically slowed fast resensitization. In contrast, alphaE262 mutations minimally affected 3-azioctanol channel block, and a half blocking concentration of 3-azioctanol did not alter the rate of ACh-induced fast desensitization. Our results indicate that position alphaE262 on muscle nAChRs contributes to an allosteric modulator site that is strongly coupled to desensitization. Occupation of this pocket by hydrophobic molecules stabilizes a desensitized state by slowing resensitization. PMID- 17910480 TI - The photosystem of Rhodobacter sphaeroides assembles with zinc bacteriochlorophyll in a bchD (magnesium chelatase) mutant. AB - A Rhodobacter sphaeroides bchD (magnesium chelatase) mutant was studied to determine the properties of its photosystem in the absence of bacteriochlorophyll (BChl). Western blots of reaction center H, M, and L (RC H/M/L) proteins from mutant membranes showed levels of 12% RC H, 32% RC L, and 46% RC M relative to those of the wild type. Tricine-SDS-PAGE revealed 52% light-harvesting complex alpha chain and 14% beta chain proteins compared to those of the wild type. Pigment analysis of bchD cells showed the absence of BChl and bacteriopheophytin (BPhe), but zinc bacteriochlorophyll (Zn-BChl) was discovered. Zn-BChl binds to light-harvesting 1 (LH1) and 2 (LH2) complexes in place of BChl in bchD membranes, with a LH2:LH1 ratio resembling that of wild-type cells under BChl limiting conditions. Furthermore, the RC from the bchD mutant contained Zn-BChl in the special pair and accessory BChl binding sites, as well as carotenoid and quinone, but BPhe was absent. Comparison of the bchD mutant RC absorption spectrum to that of Acidiphilium rubrum, which contains Zn-BChl in the RC, suggests the RC protein environment at L168 contributes to A. rubrum special pair absorption characteristics rather than solely Zn-BChl. We speculate that Zn-BChl is synthesized via the normal BChl biosynthetic pathway, but with ferrochelatase supplying zinc protoporphyrin IX for enzymatic steps following the nonfunctional magnesium chelatase. The absence of BPhe in bchD cells is likely related to Zn2+ stability in the chlorin macrocycle and consequently high resistance of Zn-BChl to pheophytinization (dechelation). Possible agents prevented from dechelating Zn BChl include the RC itself, a hypothetical dechelatase enzyme, and spontaneous processes. PMID- 17910481 TI - Chemical dissection of the APC Repeat 3 multistep phosphorylation by the concerted action of protein kinases CK1 and GSK3. AB - A crucial event in machinery controlled by Wnt signaling is the association of beta-catenin with the adenomatous polyposis coli (APC) protein, which is essential for the degradation of beta-catenin and requires the multiple phosphorylation of APC at six serines (1501, 1503, 1504, 1505, 1507, and 1510) within its repeat three (R3) region. Such a phosphorylation is believed to occur by the concerted action of two protein kinases, CK1 and GSK3, but its mechanistic aspects are a matter of conjecture. Here, by combining the usage of variably phosphorylated peptides reproducing the APC R3 region and Edman degradation assisted localization of residues phosphorylated by individual kinases, we show that the process is initiated by CK1, able to phosphorylate S1510 and S1505, both specified by non-canonical determinants. Phosphorylation of S1505 primes subsequent phosphorylation of S1501 by GSK3. In turn, phospho-S1501 triggers the hierarchical phosphorylation of S1504 and S1507 by CK1. Once phosphorylated, S1507 primes the phosphorylation of both S1510 and S1503 by CK1 and GSK3, respectively, thus completing all six phosphorylation steps. Our data also rule out the intervention of CK2 despite the presence of a potential CK2 phosphoacceptor site, S1510LDE, in the R3 repeat. S1510 is entirely unaffected by CK2, while it is readily phosphorylated even in the unprimed peptide by CK1delta but not by CK1gamma. This discloses a novel motif significantly different from non-canonical sequences phosphorylated by CK1 in other proteins, which appears to be specifically recognized by the delta isoform of CK1. PMID- 17910483 TI - Controlled particle placement through convective and capillary assembly. AB - A wide variety of methods are now available for the synthesis of colloidal particle having controlled shapes, structures, and dimensions. One of the main challenges in the development of devices that utilize micro- and nanoparticles is still particle placement and integration on surfaces. Required are engineering approaches to control the assembly of these building blocks at accurate positions and at high yield. Here, we investigate two complementary methods to create particle assemblies ranging from full layers to sparse arrays of single particles starting from colloidal suspensions of gold and polystyrene particles. Convective assembly was performed on hydrophilic substrates to create crystalline mono- or multilayers using the convective flow of nanoparticles induced by the evaporation of solvent at the three-phase contact line of a solution. On hydrophobic surfaces, capillary assembly was investigated to create sparse arrays and complex three-dimensional structures using capillary forces to trap and organize particles in the recessed regions of a template. In both methods, the hydrodynamic drag exerted on the particle in the suspension plays a key role in the assembly process. We demonstrate for the first time that the velocity and direction of particles in the suspension can be controlled to perform assembly or disassembly of particles. This is achieved by setting the temperature of the colloidal suspension above or below the dew point. The influence of other parameters, such as substrate velocity, wetting properties, and pattern geometry, is also investigated. For the particular case of capillary assembly, we propose a mechanism that takes into account the relative influences of these parameters on the motion of particles and that describes the influence of temperature on the assembly efficiency. PMID- 17910482 TI - Cyclooxygenase-2-mediated metabolism of arachidonic acid to 15-oxo eicosatetraenoic acid by rat intestinal epithelial cells. AB - Rat intestinal epithelial cells that permanently express the cyclooxygenase-2 (COX-2) gene (RIES cells) were used to investigate COX-2-mediated arachidonic acid (AA) metabolism. A targeted chiral lipidomics approach was employed to quantify AA metabolites that were secreted by the cells into the culture media. When intact RIES cells were treated with calcium ionophore A-23187 (1 microM) for 1 h, 11-(R)-hydroxyeicosatetraenoic acid (HETE) was the most abundant metabolite, followed by prostaglandin (PG) E 2, 15-(S)-HETE, 15-oxo-eicosatetraenoic acid (ETE), and 15-(R)-HETE. Incubation for a further 23 h after the calcium ionophore was removed resulted in a substantial increase in PGE 2 concentrations while HETE and 15-oxo-ETE concentrations decreased to almost undetectable levels. A similar metabolic profile was observed when RIES cells were treated with increasing concentrations of AA for 24 h. Incubation of the RIES cells with 10 microM AA revealed that maximal concentrations of 11-(R)-HETE, 15-(S)-HETE, and 15-oxo-ETE occurred after 10 min of incubation when the 15-( S)-HETE concentrations were approximately twice that of PGE 2. There was a gradual decrease in the concentrations of HETE and 15-oxo-ETE over time, whereas PGE 2 concentrations increased steadily until they reached a maximum after 24 h of incubation. The ratio of PGE 2 to 15-(S)-HETE was then approximately 20:1. 15-(S)-HETE and 15-oxo ETE concentrations declined in the cell media during prolonged incubations with pseudo-first-order rate constants of 0.0121 and 0.0073 min(-1), respectively. 15 (S)-HETE was shown to undergo metabolism primarily to 15-oxo-ETE, which was further metabolized to a glutathione (GSH) adduct. The GSH adduct of 15-oxo-ETE was further metabolized in the extracellular milieu to a cysteinylglycine adduct. Thus, we have established for the first time that 15-oxo-ETE can be formed biosynthetically from AA, that 15-(S)-HETE is its immediate precursor, and that 15-oxo-ETE forms a GSH adduct. For ionophore-A-23187-stimulated cells and at early time points for AA-stimulated cells, 11-(R)-HETE was the major eicosanoid to be secreted into the media. Adding increasing concentrations of AA to cells in culture made it possible to estimate with surprising accuracy endogenous eicosanoid production using regression analyses. Thus, after 24 h in the absence of added AA, 11-(R)-HETE and 15-(R)-HETE were estimated to be present at concentrations close to the detection limit of our very sensitive assay. These data further highlight the importance of endogenous COX-2-mediated lipid peroxidation and illustrate the necessity to monitor eicosanoid formation from endogenous stores of AA in cell culture experiments. PMID- 17910484 TI - Thermal oxidation of 6 nm aerosolized silicon nanoparticles: size and surface chemistry changes. AB - The earliest stages of thermal oxidation of 6 nm diameter silicon nanoparticles by molecular oxygen are examined using a tandem differential mobility analysis (TDMA) apparatus, Fourier-transform infrared (FTIR) spectroscopy, time-of-flight secondary ion mass spectroscopy (ToF-SIMS), and X-ray photoelectron spectroscopy (XPS). Particles are synthesized in and then extracted from a nonthermal RF plasma operating at approximately 20 Torr into the atmospheric pressure TDMA apparatus. The TDMA apparatus was used to measure oxidation-induced size changes over a broad range of temperature settings and N2-O2 carrier gas composition. Surface chemistry changes are evaluated in situ with an FTIR spectrometer and a hybrid flow-through cell, and ex situ with ToF-SIMS and XPS. Particle size measurements show that, at temperatures less than approximately 500 degrees C, particles shrink regardless of the carrier gas oxygen concentration, while FTIR and ToF-SIMS spectra demonstrate a loss of hydrogen from the particles and minimal oxide formation. At higher temperatures, FTIR and XPS spectra indicate that an oxide forms which tends toward, but does not fully reach, stoichiometric SiO2 with increasing temperature. Between 500 and 800 degrees C, size measurements show a small increase in particle diameter with increasing carrier gas oxygen content and temperature. Above 800 degrees C, particle growth rapidly reaches a plateau while FTIR and XPS spectra change little. ToF-SIMS signals associated with O-Si species also show an increase in intensity at 800 degrees C. PMID- 17910485 TI - Site-specific hydration status of an amphipathic peptide in AOT reverse micelles. AB - Reverse micelles formed by sodium bis(2-ethylhexyl) sulfosuccinate (AOT) in isooctane (IO) and water have long been used as a means to provide a confined aqueous environment for various applications. In particular, AOT reverse micelles have often been used as a template to mimic membrane-water interfaces. While earlier studies have shown that membrane-binding peptides can indeed be incorporated into the polar cavity of AOT reverse micelles where they mostly fold into an alpha-helical structure, the underlying interactions leading to the ordered conformation are however not well understood. Herein, we have used circular dichroism (CD) and infrared (IR) spectroscopies in conjunction with a local IR marker (i.e., the CN group of a non-natural amino acid, p-cyano phenylalanine) and a global IR reporter (i.e., the amide I' band of the peptide backbone) to probe the conformation as well as the hydration status of an antimicrobial peptide, mastoparan x (MPx), in AOT reverse micelles of different water contents. Our results show that at, w0=6, MPx adopts an alpha-helical conformation with both the backbone and hydrophobic side chains mostly dehydrated, whereas its backbone becomes partially hydrated at w0=20. In addition, our results suggest that the amphipathic alpha-helix so formed orients itself in such a manner that its positively charged, lysine-rich, hydrophilic face points toward the negatively charged AOT head groups, while its hydrophobic face is directed toward the polar interior of the water pool. This picture is in marked contrast to that observed for the binding of MPx to phospholipid bilayers wherein the hydrophobic surface of the bound alpha-helix is buried deeper into the membrane interior. PMID- 17910486 TI - Formation and properties of reverse micellar cubic liquid crystals and derived emulsions. AB - The structure of the reverse micellar cubic (I2) liquid crystal and the adjacent micellar phase in amphiphilic block copolymer/water/oil systems has been studied by small-angle X-ray scattering (SAXS), rheometry, and differential scanning calorimetry (DSC). Upon addition of water to the copolymer/oil mixture, spherical micelles are formed and grow in size until a disorder-order transition takes place, which is related to a sudden increase in the viscosity and shear modulus. The transition is driven by the packing of the spherical micelles into a Fd3m cubic lattice. The single-phase I2 liquid crystals show gel-like behavior and elastic moduli higher than 104 Pa, as determined by oscillatory measurements. Further addition of water induces phase separation, and it is found that reverse water-in-oil emulsions with high internal phase ratio and stabilized by I2 liquid crystals can be prepared in the two-phase region. Contrary to liquid-liquid emulsions, both the elastic modulus and the viscosity decrease with the fraction of dispersed water, due to a decrease in the crystalline fraction in the sample, although the reverse emulsions remain gel-like even at high volume fractions of the dispersed phase. A temperature induced order-disorder transition can be detected by calorimetry and rheometry. Upon heating the I2 liquid crystals, two thermal events associated with small enthalpy values were detected: one endothermic, related to the "melting" of the liquid crystal, and the other exothermic, attributed to phase separation. The melting of the liquid crystal is associated with a sudden drop in viscosity and shear moduli. Results are relevant for understanding the formation of cubic-phase-based reverse emulsions and for their application as templates for the synthesis of structured materials. PMID- 17910488 TI - Anomalous electrochemical dissolution and passivation of iron growth catalysts in carbon nanotubes. AB - Catalytically synthesized carbon nanotubes (CNTs) such as those prepared via chemical vapor deposition (CVD) contain metallic impurities including Fe, Ni, Co, and Mo. Transition metal contaminants such as Fe can participate in redox cycling reactions that catalyze the generation of reactive oxygen species and other products. Through the nature of the CVD growth process, metallic nanoparticles become encased within the CNT graphene lattice and may still be chemically accessible and participate in redox chemistry, especially when these materials are utilized as electrodes in electrochemical applications. We demonstrate that metallic impurities can be selectively dissolved and/or passivated during electrochemical potential cycling. Anomalous Fe dissolution and passivation behavior is observed in neutral (pH=6.40+/-0.03) aqueous solutions when using multiwalled CNTs prepared from CVD. Fe particles contained within these CNTs display intriguing, potential-dependent Fe redox activity that varies with supporting electrolyte composition. In neutral solutions containing dibasic sodium phosphate, sodium acetate, and sodium citrate, FeII dissolution and surface confined FeII/III redox activity are significant despite Fe being encapsulated within CNT graphene layers. However, no apparent Fe dissolution is observed in 1 M potassium nitrate solutions, suggesting that the electrolyte composition plays an important role in observing FeII dissolution, passivation, and surface confined FeII/III redox activity. Between potentials of 0 and -1.1 V versus Hg/Hg2SO4, the primary redox-active Fe species are surface FeII/III oxides/oxyhydroxides. This FeII/III surface oxide redox chemistry can be completely suppressed by passivating Fe through repeated cycling of the CNTs in supporting electrolyte. By increasing the potential to more negative values (> 1.3 V), FeII dissolution may be induced in electrolyte solutions containing acetate and phosphate and inhibited by addition of sodium benzoate, which adsorbs on exposed Fe particles, effectively passivating them. Finally, we observe that the FeII/III redox chemistry or subsequent passivation does not affect the onset of oxygen reduction at nitrogen-doped CNTs, suggesting that the surface-bound FeII species is not the primary catalytically active site for oxygen reduction in these materials. PMID- 17910487 TI - Highly specific affinities of short peptides against synthetic polymers. AB - We investigated polymer-binding 7-mer peptides that recognize differences in the polymer stereoregularity of all-purpose poly(methyl methacrylate)s (PMMAs) with simple chemical structures. Quantitative surface plasmon resonance measurements detected association/dissociation processes of the peptides against PMMA film surfaces, followed by an estimation of kinetic parameters such as association/dissociation rate constants and affinity constants. Greater association and smaller dissociation constants of the peptides were observed against a target isotactic PMMA than the structurally similar reference syndiotactic PMMA, followed by greater affinity constants against the target. A c02 peptide composed of the Glu-Leu-Trp-Arg-Pro-Thr-Arg sequence showed the greatest affinity constant (2.8x10(5) M(-1)) for the target, which was 41-fold greater than that for the reference, thus demonstrating extremely high peptide specificities. The substitution of each amino acid of the c02 peptide to Ala (Ala scanning) clearly revealed the essential amino acids for the affinity constants; the essential order was Pro5>>Thr6>Arg7>Glu1>Arg4. In fact, the shorter 4-mer peptide composed of the C-terminal Arg-Pro-Thr-Arg sequence of the c02 peptide still demonstrated strong target specificity, although the N-terminal 4-mer peptide Glu-Leu-Trp-Arg completely lost its specificity. The possible conformations modeled with Molecular Mechanics supported the significance of the Arg-Pro-Thr-Arg sequence. The thermodynamic parameters of the c02 peptide suggested an induced fit mechanism for the specific affinity. The present affinity analyses of polymer-recognizing peptides revealed significant and general information that was essential for potential applications in peptidyl nanomaterials. PMID- 17910489 TI - Microscale fish bowls: a new class of latex particles with hollow interiors and engineered porous structures in their surfaces. AB - Microscale fish bowls, hollow particles with engineered holes in their surfaces, were prepared using two different methods. In the first method, commercial latex beads suspended in water were swollen with a good solvent of the polymer, followed by freezing with liquid nitrogen and evaporation of the solvent below 0 degrees C. While one big hole was generated when the amount of solvent used for the swelling was relatively low, small holes could be produced in the outer surface of each bowl by increasing the degree of swelling. The porosity and pore structure show a similar dependence on the degree of swelling for both amorphous and semicrystalline polymers even though they are supposed to exhibit different phase behaviors during the freezing and solvent evaporation processes. In the second method, a polymer emulsion in water was prepared and then frozen with liquid nitrogen, followed by solvent evaporation below 0 degrees C. The porosity and pore structure could be controlled by adjusting the concentration of the polymer solution used to prepare the emulsion. As for encapsulation, the bowl shaped particles could be transformed back into solid beads via thermal annealing at a temperature near the glass transition temperature of the polymer or by adding a good solvent of the polymer to the colloidal suspension. In a proof-of concept experiment, microscale fish bowls were fabricated from poly(caprolactone), quickly loaded with a fluorescent dye, and sealed through thermal annealing. The encapsulated dye could then be slowly released in a phosphate buffered saline, suggesting their potential use as a new class of microscale capsules for drug delivery. PMID- 17910490 TI - Generation of stable complex gradients across two-dimensional surfaces and three dimensional gels. AB - Many chemical and biological processes are dependent on molecular gradients. We describe a new microfluidic approach that can be used to produce spatiotemporal gradients across two-dimensional surfaces and three-dimensional gels under flow free conditions. Free diffusion between dynamically replenished flow channels acting as a sink and source is utilized to give rise to stable steady-state gradient profiles. The gradient profile is dictated by the engineered design of the device's gradient-generating region. Different designs can yield both linear and non-linear gradients of varying profiles. More complex gradients can be made by juxtaposing different designs within a single gradient-generating region. By fabricating an array of designs along the gradient-generating region, different gradient profiles can be generated simultaneously, allowing for parallel analysis. Additionally, simple methods of localizing gels into microdevices are demonstrated. The device was characterized by experimentally obtained gradient profiles of fluorescent molecules that corroborated closely with a simulated finite element model. PMID- 17910492 TI - Hydration structures of bromides on cationic micelles. AB - The peripheral structures of bromides on dodecyltrimethylammonium bromide (DTAB) and hexadecyltrimethylammonium bromide (HTAB) micelles have been studied by X-ray absorption fine structure (XAFS) at the Br-K edge. The XAFS spectra indicate that water is a dominant scattering group for Br- even in these micellar solutions. However, the oscillation intensity decreases with increasing micellar concentration, suggesting that the bromides are dehydrated to some extent when they are bound to the micelles. A XAFS analysis routine gives unusually short Br O (water) distances and is inapplicable to the present systems. This comes from the structure of the first coordination shell, in which two or more scattering paths are involved. The second scattering group is obviously the head group of the surfactants forming the micelles. The detailed analysis has allowed us to estimate the hydration number of the bromides bound on the DTAB and HTAB micelles (N = ca. 4.2). The assumption that all of the bromides form direct ion-associates with the head groups causes the contradiction to the results of the XAFS analyses. This strongly implies that some of the bromides partitioned into the micelle are completely hydrated as far as their first coordination shell is concerned. Assuming that the maximum hydration number of the bromides bound to the head groups of the micelle is three, 40% of the bromides partitioned into the micelle are completely hydrated. PMID- 17910493 TI - Accessibility of the functional groups of chitosan aerogel probed by FT-IR monitored deuteration. AB - Transmission FT-IR spectroscopy allowed us to monitor the deuteration of wafers of chitosan aerogel and xerogel by D2O vapor at room temperature. The complete deuteration of the alcohol and amine groups of the aerogel (surface area 175 m2 g(-1) as measured by N2 volumetry) confirmed the high accessibility of the functional groups of the polymer. The xerogel (surface area 5 m2 g(-1)) was only partially deuterated in more severe conditions. The isotopic shift of the deuterated groups allowed us to confirm or revise some attributions of infrared bands of chitosan. PMID- 17910491 TI - Experimental approach for assessing the outcome accuracy of antibody microarray experiments. AB - An experimental strategy for quality control of antibody microarray analyses is proposed. The method utilizes proteins that are prepared for regular antibody microarray experiments. There is no need to use exogenous positive or negative reference markers and no need to determine the absolute concentration of each individual protein in the sample. Validation experiments support the basic principle of the proposed approach. This method can be a useful tool for assessing the outcome accuracy of microarray experiments. PMID- 17910494 TI - Physical properties of canola oil based polyurethane networks. AB - A new generation polyol (generation-II) with significantly higher triol content and higher hydroxyl value was synthesized from canola oil by introducing a mild solvent (ethyl acetate) and a more efficient reductive reagent (zinc) to the previous synthetic procedure (Narine, S. S.; Yue, J.; Kong, X. J. Am. Oil Chem. Soc. 2007, 84, 173-179). Polyurethane (PUR) elastomers were prepared by reacting this type of polyol with aliphatic diisocyanates. The physical and thermal properties of the PUR elastomers were studied using dynamic mechanical analysis (DMA) and differential scanning calorimetry (DSC) and compared to the elastomers made from the old generation polyol (generation-I). The concentration of elastically active network chains (nue) of the polymer networks was calculated based on rubber elasticity theory. Larger nue and narrower distribution of nue was observed in the case of the PURs prepared from the generation-II polyol. The relatively faster relaxation at higher temperature for this type of PUR elastomer, suggests a tighter cross-linked network structure by reducing the dangling chains effect. With the same OH/NCO molar ratio, the PURs prepared from the generation-II polyol showed higher glass transition temperatures (Tg), higher Young's modulus and tensile strength, and longer elongation at break. PMID- 17910496 TI - UV-patterned poly(ethylene glycol) matrix for microarray applications. AB - A versatile method to fabricate polymeric matrixes for microarray applications is demonstrated. Several different design strategies are presented where a variety of organic films, such as plastic polymers and self-assembled monolayers (SAMs) on planar silica and gold substrates, act as supports for the graft polymerization procedure. An ensemble of poly(ethylene glycol) methacrylate monomers are combined to obtain a matrix with desired properties: low nonspecific binding and easily accessible groups for postimmobilization of ligands. The free radical graft polymerization process occurs under irradiation with UV light in the 254-266 nm range, which offers the possibility to introduce patterns by means of a photomask. The arrays are created on inert and homogeneous coatings prepared either by graft polymerization of a methoxy-terminated PEG-methacrylate or self assembly of a methoxy-terminated oligo(ethylene glycol) thiol. Carboxylic acid groups, introduced in the array spots either during graft polymerization or upon wet chemical conversion of hydroxyls, grant the capability to immobilize proteins and other molecules via free amine groups. Immobilization of fluorescent species as well as biotin followed by exposure to a fluorescently labeled antibody directed toward biotin display both excellent integrity of the spots and low nonspecific binding to the surrounding framework. Beside patterns of uniform height and size, an array of spots with varying thickness (a sort of gradient) is demonstrated. Such gradient samples enable us to address critical issues regarding the mechanism(s) behind spatially resolved free radical polymerization of methacrylates. It also offers a convenient route to optimize the matrix properties with respect to thickness, loading capacity, protein diffusion/penetration, and nonspecific binding. PMID- 17910495 TI - Molecular structure and rheological properties of short-side-chain heavily glycosylated porcine stomach mucin. AB - The current accepted model for high-molecular-weight gastric mucins of the MUC family is that they adopt a polydisperse coil conformation in bulk solutions. We develop this model using well-characterized highly purified porcine gastric mucin Orthana that is genetically close to the human MUC6 type. It has short side chains and low levels of sialic acid residues and includes minute amounts of cysteine residues that, if abundant, can be responsible for the self polymerization of mucin. We have established that the mucin structure in bulk solutions corresponds to a daisy-chain random coil. Dynamic light scattering experiments probe the internal dynamics of globular subunits (individual daisies) at the approximately 9 nm length scale, whereas viscosity and light scattering measurements indicate that the size of the whole mucin chains is much larger, approximately 50 nm. The bulk viscosity (eta) scales with mucin concentration (c) in a manner similar to that found for short-side-chain synthetic comb polyelectrolytes and is characterized by a transition between semidilute (eta approximately c1/2) and entangled (eta approximately c3/2) regimes. PMID- 17910497 TI - Addition of trifluoromethyltrimethylsilane to acyl phosphonates: synthesis of TMS protected 1-alkyl-1-trifluoromethyl-1-hydroxyphosphonates and 1 aryldifluoroethenyl phosphates. AB - Addition reactions of nucleophilic CF3TMS to acyl phosphonates were investigated. Various acyl phosphonates reacted readily with CF3TMS in the presence of K2CO3 in DMF at rt to give 1-alkyl-2,2,2-trifluoro-1-trimethylsilyloxyethylphosphonate in 70-90% yields. When benzoyl phosphonates were used as starting material, after addition of CF3, the formed alcoholate undergoes phosphonate-phosphate rearrangement to form the acyl anion, followed by elimination of F- to give 1 aryldifluoroethenyl phosphates in 87-97% yields. As a representative example, vinylphosphate 6a was converted into 2,2-difluoro-1-phenylethanone 7 with 6 N HCl/EtOH/reflux or CAN/NaOH/MeOH/0 degrees C in 82-90% yields. PMID- 17910498 TI - Unprecedented rearrangement of a 4-alkoxy-5-bromoalk-2-en-1-ol to a cyclopentenone via an iso-nazarov cyclization process. AB - We report the structure determination of the product 9 of the rearrangement of the allylic alcohol 3 under very mild conditions, probably promoted by an acidic substance, and propose a reasonable mechanism for its formation. PMID- 17910499 TI - Internal azomethine ylide cycloaddition methodology for access to the substitution pattern of aziridinomitosene A. AB - Highly substituted, tethered alkyne dipolarophiles participate in the internal 2 + 3 cycloaddition with azomethine ylides generated by treatment of oxazolium salts with cyanide ion. Starting from oxazole 26, a sequence of N-methylation, cyanide addition, and electrocyclic ring opening of a 4-oxazoline intermediate affords the indoloquinone 31 in a one-pot process. A similar reaction from the protected alkynol derivative 25 affords the sensitive, but isolable, enone 32, and subsequent oxidation affords 31 and the deprotected quinone alcohol 34. Related azomethine cycloaddition methodology via intramolecular oxazolium salt formation from 43 or 46 is also demonstrated and allows the synthesis of quinone 45 and derived structures having the substitution pattern of aziridinomitosene A. Removal of the N-trityl protecting group could not be achieved without aziridine cleavage. PMID- 17910500 TI - N-silyl protecting groups for labile aziridines: application toward the synthesis of N-H aziridinomitosenes. AB - Hindered N-silylamines were examined for their utility to serve as protecting groups for the labile aziridine nitrogen found within the highly sensitive aziridinomitosene framework. tert-Butyldiphenylsilyl and modified tert butyldiphenylsilyl groups were the most resistant to nitrogen-silicon bond cleavage under various reaction conditions and were thus employed in transformations relevant to aziridinomitosene synthesis. The N-silylaziridines 7a, 21a, and 21b underwent azomethine ylide cycloaddition and afforded, upon deprotection, the N-H aziridine 24 in 18-32% overall yield for the three steps. PMID- 17910501 TI - Total synthesis of streptonigrone. AB - A total synthesis of streptonigrone, 1, is described, which incorporates a one step synthesis of substituted pyridones devised in our laboratory. Other aspects of the synthesis that differentiate the present approach from previous ones are the use of a Conrad-Limpach reaction, rather than the customary Friedlander methodology, to assemble the quinoline segment of 1, and the implementation of an anionic sequence for the functionalization of a key pyridone intermediate. PMID- 17910502 TI - A remarkable accelerating effect of Ag-salt on intramolecular cyclization of o-(1 alkynyl)benzenesulfonamides. AB - Herein, we report transition metal-catalyzed intramolecular cyclization of o-(1 alkynyl)benzenesulfonamides to afford 3-substituted benzothiazines regioselectively via a C-N bond forming reaction and Cu-catalyzed sequential C-N and C-C bond formation leading to the corresponding 3,4-disubstituted derivatives. PMID- 17910503 TI - Efficient and practical synthesis of 4(5)-aryl-1H-imidazoles and 2,4(5)-diaryl-1H imidazoles via highly selective palladium-catalyzed arylation reactions. AB - 4(5)-aryl-1H-imidazoles can be efficiently and selectively prepared by PdCl2(dppf)-catalyzed Suzuki-Miyaura reaction of commercially available 4(5) bromo-1H-imidazole with arylboronic acids under phase-transfer conditions. On the other hand, N-unprotected 4(5)-aryl-1H-imidazoles can undergo highly selective Pd(OAc)2-catalyzed and CuI-mediated direct C-2-arylation with a variety of aryl bromides and iodides under base-free and ligandless conditions to produce 2,4(5) diaryl-1H-imidazoles in modest to good yields. No N-arylation byproducts are observed under the experimental conditions used to prepare 2,4(5)-diaryl-1H imidazoles. PMID- 17910505 TI - Enantioselective synthesis of the alcohol moiety of dolabriferol. AB - Dolabriferol is a marine polypropionate characterized by an unusual noncontiguous carbon backbone. The two polypropionate subunits are linked by an ester function. The protected alcohol moiety of dolabriferol was synthesized via the enzymatic desymmetrization of meso-(anti-anti)-2,4-dimethyl-1,3,5-pentanetriol. PMID- 17910506 TI - Efficient synthesis of a new structural phenanthro[9,10,3',4']indolizidine starting from pyrrole. AB - A new structural phenanthroindolizidine, 2,3,6,7 tetramethoxyphenanthro[9,10,3',4']indolizidine, has been synthesized efficiently from pyrrole. An important feature of this synthesis is that intramolecular oxidative coupling and rearrangement of 6,7-bis(3,4-dimethoxyphenyl)-8-methoxy 1,2,3-trihydroindolizin-5-one by using VOF3 and TFA have been achieved in one pot. PMID- 17910507 TI - Selective synthesis of fluorinated furan derivatives via AgNO3-catalyzed activation of an electronically deficient triple bond. AB - The transition metal-catalyzed direct activation of electron deficient triple bonds was investigated by using the combined electron withdrawing effects of two fluorine atoms to modulate the electronic density of the triple bond. With use of catalytic amounts of AgNO3 (10 mol %) the synthesis of substituted 3,3-difluoro 4,5-dihydrofurans from gem-difluorohomopropargyl alcohols occurred in excellent NMR yields. Treatment of these dihydrofurans with SiO2 or Pd/H2 yielded the corresponding 3-fluorinated furans and 3,3-difluorotetrahydrofurans. PMID- 17910504 TI - Electrophilic chemistry of thia-PAHs: stable carbocations (NMR and DFT), S alkylated onium salts, model electrophilic substitutions (nitration and bromination), and mutagenicity assay. AB - First examples of stable carbocations are reported from several classes of thia PAHs with four fused rings, namely, benzo[b]naphtho[2,1-d]thiophene (1) and its 3 methoxy derivative (2), phenanthro[4,3-b]thiophene (3) and its 7-methoxy (4), 10 methoxy (5), and 9-methoxy (6) derivatives, phenanthro[3,4-b]thiophene (7) and its 7-methoxy (8) and 9-methoxy (9) derivatives, and 3-methoxybenzo[b]naphtha[1,2 d]thiophene (11). In several cases, the resulting carbocations were also studied by GIAO-DFT. Charge delocalization modes in the resulting carbocations were probed. A series of S-alkylated onium tetrafluoroborates, namely, 1Me+, 1Et+, 2Et+, and 7Me+ (from 1, 2, and 7), 10Me+ and 10Et+ (from benzo[b]naphtha[1,2 d]thiophene 10), 12Me+ and 12Et+ (from phenanthro[3,2-b][1]benzothiophene 12), 13Me+ (from 3-methoxyphenanthro[3,2-b]benzothiophene 13), 14Me+ (from phenanthro[4,3-b][1]benzothiophene 14), and 15Me+ (from 3-methoxyphenanthro[4,3 b][1]benzothiophene 15), were synthesized. PAH-sulfonium salts 1Me+, 1Et+, 10Me+, 10Et+, 12Me+, and 14Me+ proved to be efficient akylating agents toward model nitrogen nucleophile receptors (imidazole and azaindole). Facile transalkylation to model nucleophiles (including guanine) is also supported by favorable reaction energies computed by DFT. Ring opening energies in thia-PAH-epoxides from 1, 3, and 7 and charge delocalization modes in the resulting carbocations were also evaluated. The four-ring-fused thia-PAHs 1, 2, 3, 4, 5, 7, 8, and 11 are effectively nitrated under extremely mild conditions. Nitration regioselectivity corresponds closely to protonation under stable ion conditions. Bromination of 4 and 6 is also reported. Comparative mutagenicity assays (Ames test) were performed on 1 versus 1NO2, 5 versus 5NO2, and 11 versus 11NO2. Compound 5NO2 was found to be a potent direct acting mutagen. PMID- 17910509 TI - Light-induced off-flavor development in cloudy apple juice. AB - Cloudy apple juice has been found to develop off-flavors during storage in daylight. The development of off-flavors and volatile compounds was monitored in reconstituted juice prepared from 'Golden Delicious' and 'Fuji' apple concentrates stored in glass bottles under fluorescent light (3000 lx, 8 degrees C). A strong metallic off-flavor was formed by photooxidation. A major contributor to the off-flavor was identified as 1-octen-3-one by gas chromatography-olfactometry. In addition, six volatile compounds, pentanal, 2 methyl-1-penten-3-one, hexanal, (E)-2-heptenal, 6-methyl-5-hepten-2-one, and (E) 2-octenal, increased significantly after light exposure and could contribute to the off-flavor. Except for pentanal and hexanal, these volatiles were found only after light exposure. Higher levels of volatiles were observed in juice from 'Golden Delicious' apples than in juice from 'Fuji' apples, and this difference was consistent with higher levels of suspended solids. When the suspended solids were removed by centrifugation, the development of volatiles on exposure to light was reduced significantly. PMID- 17910508 TI - Trifluoromethylation of salicyl aldimines. AB - A method for the nucleophilic trifluoromethylation of salicyl aldimines, which do not contain an activating group at the nitrogen atom, has been described. The reaction proceeds through the initial generation of intramolecular boron complex followed by interaction with Me3SiCF3 activated by sodium acetate. PMID- 17910510 TI - Hemoglobin-mediated lipid oxidation and compositional characteristics of washed fish mince model systems made from cod (Gadus morhua), herring (Clupea harengus), and salmon (Salmo salar) muscle. AB - The use of washed cod light muscle minces in mechanistic studies of hemoglobin (Hb)-mediated fish lipid oxidation has largely increased in the past 5 years. Although cod light muscle has a low level of intrinsic lipid oxidation catalysts, a prerequisite for a good oxidation model system, we believe it cannot fully mimic the oxidation kinetics taking place in other fish species being more susceptible to lipid oxidation. The aim of this study was to systematically investigate whether washed mince model systems useful in Hb-mediated oxidation studies could be prepared also from herring (Clupea harengus) and salmon (Salmo salar) light muscles. The kinetics of oxidation in the washed models was measured during ice storage (+/-Hb), and the results were related to compositional differences. Minces from cod, herring, and salmon light muscles were washed 3 times with 3 volumes of water and buffer. A 20 microM portion of Hb and 200 ppm streptomycin was then added, followed by adjustment of pH and moisture to 6.3 and 86%, respectively. Samples with or without Hb were then stored on ice, and oxidation was followed as peroxide value (PV), rancid odor, redness (a*) loss and yellowness (b*). Prior to storage, all minces and models were also analyzed for total lipids, fatty acids, alpha-tocopherol, proteins, Hb, Fe, Cu, and Zn. Hb mediated lipid oxidation appeared within 2 days on ice in all models. Small differences in the oxidation rates ranked the models as herring > cod > salmon. These differences were ascribed to more preformed peroxides and trace elements in the herring model, and more antioxidants in the salmon model. Controls, without Hb, stayed stable in all cases except herring, where a very slight oxidation appeared, especially if the herring raw material had been prefrozen. In conclusion, fattier fish like dark muscle species and salmonoids are useful for making washed mince model systems and would be a better choice than cod if there is an interest in the oxidation kinetics of such species. PMID- 17910511 TI - Protein changes in the albedo of citrus fruits on postharvesting storage. AB - In this work, major protein changes in the albedo of the fruit peel of Murcott tangor (tangerine x sweet orange) during postharvest ageing were studied through 2D PAGE. Protein content in matured on-tree fruits and in fruits stored in nonstressing [99% relative humidity (RH) and 25 degrees C], cold (99% RH and 4 degrees C), and drought (60% RH and 25 degrees C) conditions was initially determined. Protein identification through MS/MS determinations revealed in all samples analyzed the occurrence of manganese superoxide dismutase (Mn SOD), actin, ATP synthase beta subunit (ATPase), citrus salt-stress associated protein (CitSap), ascorbate peroxidase (APX), translationally controlled tumor protein (TCTP), and a cysteine proteinase (CP) of the papain family. The latter protein was identified in two different gel spots, with different molecular mass, suggesting the simultaneous presence of the proteinase precursor and its active form. While Mn SOD, actin, ATPase, and CitSap were unchanged in the assayed conditions, TCTP and APX were downregulated during the postharvest ageing process. Ageing-induced APX repression was also reversed by drought. CP contents in albedo, which were similar in on- and off-tree fruits, were strongly dependent upon cold storage. The active/total CP protein ratio significantly increased after cold exposure. This proteomic survey indicates that major changes in protein content in the albedo of the peel of postharvest stored citrus fruits are apparently related to the activation of programmed cell death (PCD). PMID- 17910512 TI - Origin and thermodynamic properties of the instability of synthetic azo colorants in gum arabic solutions. AB - The instability of some industrially important synthetic azo colorants, including sunset yellow, azorubine, and allura red, toward gum arabic in aqueous solution has been a long-standing problem for the beverage and confectionery industries. Precipitation of these colorants causes the deterioration of product appearance and properties. This work examines the origin and nature of the problem by analysis of the precipitate and thermodynamic studies of gum arabic-colorant interactions using isothermal titration calorimetry (ITC). The presence of divalent alkaline earth metals in gum arabic samples, that is, calcium and magnesium, is shown to be responsible for the precipitation of the azo colorants. There is no direct interaction between gum arabic and the colorant molecules, and the precipitate is formed likely due to the mediation/bridging by the divalent cations. The thermodynamic knowledge gained from the ITC studies, for example, binding affinity, stoichiometry, and enthalpy, enables interpretation of many industrial observations. PMID- 17910513 TI - Lyngbyastatins 5-7, potent elastase inhibitors from Floridian marine cyanobacteria, Lyngbya spp. AB - Three new analogues of dolastatin 13, termed lyngbyastatins 5-7 ( 1- 3), were isolated from two different collections of marine cyanobacteria, Lyngbya spp., from South Florida. Their planar structures were deduced by a combination of NMR techniques, and the absolute configurations were established by modified Marfey's analysis of the acid hydrolyzates. The related cyclodepsipeptide somamide B ( 4), previously reported from a Fijian cyanobacterium, has also been found in one of the extracts, and its absolute stereochemistry was unambiguously assigned for the first time. Compounds 1- 4 were found to selectively inhibit elastase over several other serine proteases, with IC50 values for porcine pancreatic elastase ranging from 3 to 10 nM. PMID- 17910514 TI - A library of 3-aryl-4,5-dihydroisoxazole-5-carboxamides. AB - Parallel solution phase methods for the preparation of a 72-membered 3-aryl-4,5 dihydroisoxazole-5-carboxamide library is reported. The reaction order (nitrile oxide 1,3-dipolar cycloaddition followed by amide formation, or vice versa) was investigated both experimentally and computationally to determine which route would result in the highest yields, minimize purification efforts, and give higher 1,3-dipolar cycloaddition regioselectivity. Automated preparative HPLC was used to purify the final products to >or=90% purity on a 10+ mg scale. PMID- 17910515 TI - Ionic liquids in separations. AB - Ionic liquids are liquids composed completely of ions. In the past two decades, ionic liquids have been widely used as "green solvents" replacing traditional organic solvents for organic synthesis and catalysis. In addition, ionic liquids are playing an increasingly important role in separation science. In this Account, the application of ionic liquids in all areas of separation science including extractions, gas chromatography, and supported liquid membrane processes are highlighted. PMID- 17910517 TI - Atorvastatin: pharmacological characteristics and lipid-lowering effects. AB - By inhibiting 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductase, the statins reduce hepatocyte cholesterol levels, which results in up-regulation of low-density lipoprotein (LDL) receptors and, consequently, increased clearance of LDL-cholesterol (LDL-C) from the plasma. Structural differences among the available statins partially account for differences in their capacity to inhibit HMG-CoA reductase and their lipid-lowering efficacy, and for variability in other biological properties, such as their pharmacokinetic characteristics and their tolerability and propensity to interact with other drugs. In terms of pharmacokinetic properties, the synthetic (type II) HMG-CoA analogue atorvastatin exhibits a number of characteristics that are different to those of other members of the class, including a longer plasma half-life and metabolites that have an ability to inhibit HMG-CoA reductase equivalent to that of the parent drug. These characteristics are postulated to be responsible for a more prolonged inhibition of HMG-CoA reductase, and, hence, for the greater efficacy of atorvastatin in decreasing total and LDL-C levels relative to other statins (with the exception of rosuvastatin) noted in clinical trials in patients with dyslipidaemias. From the available clinical trial data, atorvastatin can be considered one of the most effective statins, not only by taking into account its effects on LDL-C and ability to meet recommended treatment guidelines for this parameter, but also its effect on triglyceride levels and capacity to modify lipoprotein composition in a non-atherogenic manner. Clinical studies with atorvastatin have also shed some light on the question as to whether it is better to focus on obtaining maximal reduction of LDL-C in at-risk patients or on cardiovascular outcomes. Cardiovascular event rates have been shown to be substantially lower in patients attaining LDL-C levels between 1.0 and 1.6 mmol/L (40-60 mg/dL) or < or =1.0 mmol/L (< or =40 mg/dL) compared with higher levels (>2.1-2.6 mmol/L [>80-100 mg/dL]). This finding reinforces the update of the National Cholesterol Education Programme's clinical practice guidelines, which recommend LDL-C levels <2.6 mmol/L (100 mg/dL) to be the goal of antihyperlipidaemic drug therapy in high risk patients with CHD, with an optional therapeutic target of <1.8 mmol/L (70 mg/dL) in patients at very high risk. PMID- 17910518 TI - Effects of atorvastatin on the different phases of atherogenesis. AB - Molecular differences among the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) give rise to some important differences in their properties, including their anti-atherogenic and anti-inflammatory actions (among the so-called pleiotropic effects) - differences that may help to account for variation in clinical efficacy and safety among the available drugs of this class. The question of whether the clinical benefit of statins such as atorvastatin in reducing cardiovascular events in individuals with elevated cholesterol levels results from direct anti-atherogenic effects in addition to cholesterol-lowering-dependent effects cannot be conclusively answered at present. However, the available evidence suggests that these actions should be considered further, especially in some clinical situations such as acute coronary syndrome where an anti-inflammatory effect could conceivably have a greater role. In the case of atorvastatin, various direct anti-atherogenic effects have been demonstrated. These effects include modification of endothelial dysfunction, inflammatory processes and lipid oxidation, and a possible direct effect on the composition of atheromatous plaques, which together may have a positive influence on the development of atherosclerosis and its subsequent progression (e.g. on the reduction of carotid intima media thickness and regression of atheromas noted in studies with intensive atorvastatin therapy [80 mg/day]). In terms of its effects on endothelial function, improvements in flow-mediated endothelium-dependent vasodilatation have been observed as early as 2 weeks after starting atorvastatin treatment. This effect does not appear to be quantitatively correlated with lowering of low-density lipoprotein-cholesterol (LDL-C) as greater improvements in endothelial function versus ezetimibe/simvastatin have been noted with atorvastatin despite comparable reductions in LDL-C. Atorvastatin has also been shown to reduce levels of the inflammatory marker C-reactive protein; in comparative studies, this effect proved to be superior to that of simvastatin or pravastatin and equivalent to that of rosuvastatin. In other studies, atorvastatin has been found to inhibit the in vitro oxidation of LDL - an effect that appears to be due mainly its active hydroxy metabolite - and to reduce various oxidative stress markers in hypercholesterolaemic patients. In addition, there is evidence that atorvastatin is able to modify the composition of atherosclerotic plaques and their inflammatory status via a series of effects, mostly involving tissue factors. PMID- 17910519 TI - Atorvastatin efficacy in the primary and secondary prevention of cardiovascular events. AB - Atorvastatin has been extensively studied in the primary and secondary prevention of cardiovascular events, and may have some clinical advantages over various other statins in these respects. The principal primary prevention study of atorvastatin, ASCOT-LLA (Anglo-Scandinavian Cardiac Outcomes Trial-Lipid Lowering Arm), revealed that atorvastatin reduced the relative risk of primary coronary heart disease (CHD) events by 36% (p = 0.0005) compared with placebo in patients with hypertension. Much published data confirm the secondary preventive benefits of atorvastatin in various clinical settings. The IDEAL (Incremental Decrease in End Points Through Aggressive Lipid Lowering) and TNT (Treating to New Targets) trials demonstrate the preventive efficacy of atorvastatin in patients with stable CHD. Relative to simvastatin (in the IDEAL trial) and low-dosage atorvastatin (in the TNT trial), intensive atorvastatin therapy (80 mg/day) reduced the risk of nonfatal myocardial infarction (MI) by 17-22% (p < or = 0.02). Furthermore, the ALLIANCE (Aggressive Lipid-Lowering Initiation Abates New Cardiac Events) and GREACE (GREek Atorvastatin and Coronary-heart-disease Evaluation) trials highlight the benefits of atorvastatin in the 'real world' setting in patients with stable CHD. Compared with 'usual' care, atorvastatin reduced the risk of nonfatal MI by 47-59% (p < or = 0.0002).Moreover, the MIRACL (Myocardial Ischemia Reduction with Aggressive Cholesterol Lowering), PROVE-IT (PRavastatin Or atorVastatin Evaluation and Infection Therapy) and IDEAL-ACS (Acute Coronary Syndromes) studies outline the benefits of high-dosage atorvastatin therapy started within 24-96 hours, 10 days or 2 months, respectively, of an acute coronary syndrome. Relative to placebo, pravastatin and simvastatin, atorvastatin reduced the risk of death or major cardiovascular events by 16-18% (p < or = 0.048). In patients undergoing revascularisation procedures, the AVERT (Atorvastatin VErsus Revascularisation Treatment) study revealed that 18 months' administration of atorvastatin 80 mg/day was at least as effective as angioplasty plus usual care in reducing the risk of ischaemic events in low-risk patients with stable coronary artery disease. Furthermore, the ARMYDA (Atorvastatin for Reduction in MYocardial DAmage during angioplasty) and ARMYDA-3 trials showed that 7 days' administration of atorvastatin 40 mg/day before coronary intervention significantly reduced the risks of periprocedural myocardial damage (ARMYDA), postprocedural MI (p = 0.025; ARMYDA) and atrial fibrillation (p = 0.003; ARMYDA-3) versus placebo. In addition, it has been reported that C-reactive protein levels and the combined incidence of cardiovascular events (death, MI and target segment revascularisation during the 6-month follow-up) were significantly higher in coronaropathic patients undergoing non-surgical revascularisation procedures (stent implantation) not receiving statin therapy compared with those treated with atorvastatin (80mg). Overall, therefore, the marked efficacy of atorvastatin in the primary and secondary prevention of cardiovascular events underscores the pivotal place that this statin has in general cardiovascular disease management, and suggests even greater potential clinical utility for the drug in some clinical settings. PMID- 17910521 TI - Atorvastatin: its clinical role in cerebrovascular prevention. AB - An association between hypercholesterolaemia and ischaemic stroke has not yet been clearly defined by observational studies. In clinical trials, however, cholesterol-lowering treatments appear to consistently reduce stroke risk. Data are now available from various primary prevention studies - ALLHAT-LLT (Antihypertensive and Lipid-Lowering treatment to prevent Heart Attack, Lipid Lowering Therapy), ASCOT-LLA (Anglo-Scandinavian Cardiac Outcomes Trial, Lipid Lowering Arm), CARDS (Collaborative Atorvastatin Diabetes Study, WOSCOPS (West of Scotland COronary Prevention Study) - and secondary prevention studies - 4S (Scandinavian Simvastatin Survival Study), CARE (Cholesterol and Recurrent Events), GREACE (GREek Atorvastatin and Coronary-heart-disease Evaluation), HPS (Heart Protection Study), LIPID (Long-term Intervention with Pravastatin in Ischaemic Disease), MIRACL (Myocardial Ischemia Reduction with Aggressive Cholesterol Lowering), SPARCL (Stroke Prevention by Aggressive Reduction in Cholesterol Levels), TNT (Treating to New Targets) - confirming the ability of statins to reduce stroke risk. Regarding primary prevention, post hoc analyses showed pravastatin reduced the relative risk of stroke by 9-11% (not statistically significant) in the ALLHAT-LLT and WOSCOPS trials, whereas atorvastatin reduced this risk by 27-48% in the ASCOT-LLA (p = 0.024) and CARDS trials. It remains to be established in prospective studies whether cholesterol lowering is effective in the primary prevention of stroke. Regarding secondary prevention, in five placebo-controlled studies (4S, CARE, HPS, LIPID, MIRACL) involving a total of >40 000 patients with coronary heart disease (CHD), statin therapy reduced the relative risk of fatal or nonfatal stroke by 19-50% (p < or = 0.048); the largest decrease was produced by atorvastatin in the MIRACL study ( 50%, p = 0.045). In addition, high-dosage atorvastatin reduced stroke risk by 25% (p = 0.02) relative to lower-dosage therapy in the TNT trial, and by 47% (p = 0.034) relative to 'usual' care in the GREACE study. A post hoc analysis of data for 3280 HPS study participants who had had a previous stroke revealed that simvastatin reduced major vascular events by 20% (p = 0.001).The SPARCL study assessed the secondary preventive efficacy of atorvastatin versus placebo in 4731 patients with a history of stroke or transient ischaemic attack (TIA), but without CHD. Atorvastatin reduced the adjusted relative risk of fatal or nonfatal stroke by 16% (p = 0.03), and that of fatal stroke alone by 43% (p = 0.03). Among secondary study endpoints, atorvastatin reduced the relative risks of stroke and TIA (-23%; p < 0.001), TIA alone (-26%; p = 0.004), and ischaemic stroke (-22%; p = 0.01). Overall, SPARCL study findings suggest that intensive atorvastatin therapy should be started immediately after a stroke or TIA. In summary, atorvastatin has developed a well defined role in the primary and secondary prevention of cerebrovascular disease, and appears to have a particularly prominent place in preventing such disease in CHD patients, and in the post stroke and post-TIA setting in patients without CHD. PMID- 17910520 TI - Atorvastatin efficacy in the prevention of cardiovascular events in patients with diabetes mellitus and/or metabolic syndrome. AB - Several large-scale clinical trials have assessed the efficacy of atorvastatin in the primary and secondary prevention of cardiovascular events in patients with diabetes mellitus and/or metabolic syndrome. In primary prevention, CARDS (Collaborative Atorvastatin Diabetes Study) showed that atorvastatin 10 mg/day (vs placebo) reduced relative risk of the composite primary endpoint (acute coronary heart disease [CHD] events, coronary revascularisation, or stroke) by 37% (p = 0.001). This decrease was similar to decreases in major cardiovascular events in the ASCOT-LLA (Anglo-Scandinavian Cardiac Outcomes Trial-Lipid Lowering Arm) trial and HPS (Heart Protection Study). However, in CARDS, atorvastatin efficacy was evident as early as 6 months after starting treatment, whereas in HPS, simvastatin efficacy was noticeable only from about 15-18 months after starting treatment. In the ASCOT-LLA trial, in 2226 hypertensive diabetic patients without previous cardiovascular disease, atorvastatin (vs placebo) reduced the relative risk of all cardiovascular events and procedures by 25% (p = 0.038). In secondary prevention, substudies of the GREACE (GREek Atorvastatin and Coronary-heart-disease Evaluation), TNT (Treating to New Targets) and PROVE-IT (PRavastatin Or atorVastatin Evaluation and Infection Therapy) trials reported results for the approximately 15-25% of study participants who had diabetes. In the GREACE substudy, atorvastatin (vs physicians' standard care) significantly reduced the relative risk of total mortality by 52% (p = 0.049), coronary mortality by 62% (p = 0.042), coronary morbidity by 59% (p < 0.002) and stroke by 68% (p = 0.046). In the TNT substudy, incidence of the primary endpoint was significantly lower in diabetic patients treated with atorvastatin 80 mg/day rather than 10 mg/day (13.8% vs 17.9%; relative risk 0.75; p = 0.026). In the PROVE-IT substudy, a significantly lower incidence of acute cardiac events was reported for atorvastatin versus pravastatin recipients (21.1% vs 26.6%; p = 0.03) and, therefore, an absolute risk reduction of 5.5% was associated with atorvastatin therapy. ASPEN (Atorvastatin Study for Prevention of coronary heart disease Endpoints in Non-insulin-dependent diabetes mellitus) - a mixed primary and secondary prevention trial in diabetic patients - found that a 29% lower low density lipoprotein-cholesterol level was seen with atorvastatin than placebo at endpoint (p < 0.0001); however, the reduction in composite primary endpoint of major cardiovascular events (cardiovascular mortality, nonfatal major cardiovascular event or stroke, and unstable angina requiring hospitalisation) with atorvastatin (13.7% vs 15.0% with placebo), and reduction in acute myocardial infarction relative risk of 27% with atorvastatin were not statistically significant. In CHD patients with metabolic syndrome (n = 5584) in a sub-analysis of the TNT trial, intensive versus lower-dosage atorvastatin therapy reduced the relative risk of major cardiovascular and cerebrovascular events by 29% (p < 0.0001). The analysis also revealed that CHD patients with, rather than those without, metabolic syndrome had a 44% greater level of absolute cardiovascular risk, thus clearly underscoring the clinical feasibility of administering intensive lipid-lowering therapy to CHD patients with metabolic syndrome. In summary, several patient populations, from definitive, large-scale studies, are now available to corroborate the integral place of atorvastatin--in line with various regional and internationally accepted disease management guidelines--in the primary and secondary prevention of cardiovascular events in patients with diabetes and/or metabolic syndrome. PMID- 17910522 TI - Atorvastatin: a safety and tolerability profile. AB - Extensive data are available on the safety of atorvastatin from randomised clinical trials, postmarketing analyses and reports to regulatory agencies. Atorvastatin is generally well tolerated across the range of therapeutic dosages, with the exception of a slightly higher rate of liver enzyme elevations with atorvastatin 80 mg/day which does not appear to confer an increased risk of clinically important adverse events. Unlike simvastatin, atorvastatin is associated with a low incidence of muscular toxicity. It is not associated with neurological, cognitive or renal adverse effects and does not require dosage adjustment in patients with renal dysfunction, due to its favourable pharmacokinetic profile, which is unique among the statins. In patients aged > or =65 years, atorvastatin is well tolerated with no dose-dependent increase in adverse events up to the maximum daily dosage of 80 mg/day. Thus, atorvastatin is a safe and well tolerated statin for use in a wide range of patients. PMID- 17910523 TI - Healing of segmental bone defects by direct percutaneous gene delivery: effect of vector dose. AB - Previous studies have demonstrated the ability of direct adenoviral BMP-2 (Ad.BMP 2) gene delivery to enhance bone repair. Nevertheless, in studies using a rat segmental defect model, it has not proved possible to achieve reliably full osseous union in all animals. To address this issue, we evaluated the effect of vector dose on healing. Critical-size defects were created in the right femora of 27 Sprague-Dawley rats. The defects received a single, intralesional, percutaneous injection of 2.7 x 10(7) (low dose), 2.7 x 10(8) (medium dose), or 2.7 x 10(9) (high dose) plaque-forming units of Ad.BMP-2. After 8 weeks, femora were evaluated by X-ray, dual-energy X-ray absorptiometry, microcomputed tomography (microCT), and histology. The high dose of vector bridged 100%, the medium dose 11%, and the low dose 25% of the defects, as evaluated by X-ray and microCT imaging. Bone mineral content and bone volume of the defects receiving the high dose of vector were significantly higher than those of both groups receiving lower doses. Histologically, defects treated with the high dose were filled by trabecular bone and small amounts of cartilage, whereas large areas of fibrous tissue and cartilage remained in the defects receiving lower doses. However, the newly formed bone lacked the structural organization of native bone, suggesting that further maturation is necessary. PMID- 17910524 TI - Overexpression of E2F1 in clear cell renal cell carcinoma: a potential impact of erroneous regulation by thyroid hormone nuclear receptors. AB - We show here that the promoter of E2F1 gene, encoding one of the key regulators of cell proliferation, is overly active in the presence of low amounts of triiodothyronine (T3) and in the presence of mutant thyroid hormone receptor. We also show that T3-thyroid hormone receptor pathway of regulation of molecular processes is disturbed in clear cell renal cell carcinoma (ccRCC) on several levels, including overexpression of thyroid hormone receptors and the disturbance of their binding to DNA and to the hormone. In comparison to the cancer-free kidneys and peritumoral respective control tissues, E2F1 mRNA and protein levels are significantly increased in cancer tissues. A significant correlation between E2F1 mRNA and protein levels has been found in both control types and ccRCCs. No correlation was observed between the amount of E2F1 mRNA and the amount of thyroid hormone receptors or their DNA or T3 binding activity, suggesting that the function of thyroid hormone receptors could be markedly disturbed in both tumor and peritumoral cells. In summary, we show that ccRCC is characterized by the overexpression of E2F1, which is likely a result of a deregulated control of T3-dependent molecular processes. PMID- 17910525 TI - Value of repeated fine needle aspirations of the thyroid: an analysis of over ten thousand FNAs. AB - OBJECTIVE: Fine needle aspiration (FNA) has been accepted as the diagnostic method of choice in the initial evaluation of thyroid nodules. However, the value of repeated FNAs in the long-term follow-up of lesions initially diagnosed as benign is being questioned. Do the findings on initial FNA really spare patients thyroidectomy or do they only postpone it? The purpose of the present study is our attempt to answer this question. DESIGN: Retrospective review of cytology reports of patients who underwent thyroidal FNAs at Washington Hospital Center from January 1998 through April 2006. All statistical analyses were done using the statistical package Splus. MAIN OUTCOME: Patients who had thyroid nodules diagnosed as benign on FNA performed at our institution had a 90% probability of a benign diagnosis (with 95% confidence interval [0.87, 0.92]), when they underwent surgery. When the benign cytologic diagnosis was confirmed on a repeat aspiration, this probability increased to 98% (with 95% confidence interval [0.94, 1.0]). CONCLUSIONS: Repeated thyroidal FNAs yielding benign diagnoses are nearly always accurate (98%), and therefore the patients can be followed safely without undergoing surgery, unless an unfavorable clinical change occurs. PMID- 17910527 TI - Chemokine orchestration of autoimmune thyroiditis. AB - Chemokines are low-molecular-weight proteins that attract leukocytes and other cell types, via interaction with G protein-coupled receptors. Chemokines control leukocyte migration not only during inflammatory processes, but also throughout ontogeny and differentiation of lymphoid tissues. They have been involved in the pathogenesis of numerous diseases, such as human immunodeficiency virus infection, allergy, atherosclerosis, cancer, and autoimmunity. The number of studies focusing on chemokine biology is expanding exponentially. For example, searching PubMed for the terms "thyroid" and "chemokine" retrieved 1 article in 1980s, 18 articles in 1990s, and 81 articles from 2000 to July 2007. This review will focus on studies analyzing the role of chemokine in autoimmune thyroiditis (Graves' disease and Hashimoto's thyroiditis), performed in both patients and experimental animals. The goal is to emphasize how a better understanding of chemokine biology has advanced our knowledge of the pathogenesis of autoimmune thyroiditis. PMID- 17910526 TI - Contrasting roles of IFN-gamma in murine models of autoimmune thyroid diseases. AB - Interferon-gamma (IFN-gamma), a prototypic proinflammatory cytokine produced by several different cell types, including the Th1 subset of CD4(+) T cells, plays an important role in inflammation and autoimmune diseases. This review focuses on the varied and often contrasting roles of IFN-gamma in three murine models of autoimmune thyroid disease, experimentally induced autoimmune thyroiditis, the model of iodine-induced spontaneous autoimmune thyroiditis in NOD.H-2h4 mice and several different murine models of Graves' disease. PMID- 17910528 TI - Interview with Bruce Eldridge, PhD. Interview by Vicki Glaser. PMID- 17910529 TI - Multiple testing issues in discriminating compound-related peaks and chromatograms from high frequency noise, spikes and solvent-based noise in LC-MS data sets. AB - Liquid Chromatography--Mass Spectrometry (LC-MS) is a powerful method for sensitive detection and quantification of proteins and peptides in complex biological fluids like serum. LC-MS produces complex data sets, consisting of some hundreds of millions of data points per sample at a resolution of 0.1 amu in the m/z domain and 7000 data points in the time domain. However, the detection of the lower abundance proteins from this data is hampered by the presence of artefacts, such as high frequency noise and spikes. Moreover, not all of the tens of thousands of the chromatograms produced per sample are relevant for the pursuit of the biomarkers. Thus in analysing the LC-MS data, two critical pre processing issues arise. Which of the thousands of the: 1. chromatograms per sample are relevant for the detection of the biomarkers?, and 2. signals per chromatogram are truly compound-related? Each of these issues involves assessing the significance (deviation from noise) of multiple observations and the issue of multiple comparisons arises. Current methods disregard the multiplicity and provide no concrete threshold for significance. However, with such procedures, the probability of one or more false-positives is high as the number of tests to be performed is large, and must be controlled. Realizing that the cut-offs for declaring a chromatogram (or a signal) to be compound-related can hugely influence which proteins are detected, it seems natural to define thresholds that are neither arbitrary nor subjective. We suggest the choice of thresholds guided by the critical aim of controlling the False Discovery Rate (FDR) in multiple hypotheses testing for significance over a large set of features produced per sample. This involves the use of the regression diagnostics to characterize the signals of a chromatogram (e.g. as outliers or influential) and to suggest suitable tests statistics for the multiple testing procedures (MTP) for discriminating noise and spikes from true signals. The role of the Generalized Linear Models (GLM) in this MTP is investigated. The method is applied to LC-MS datasets from trypsin-digested serum spiked with varying levels of horse heart cytochrome C (cytoc). PMID- 17910531 TI - Super learner. AB - When trying to learn a model for the prediction of an outcome given a set of covariates, a statistician has many estimation procedures in their toolbox. A few examples of these candidate learners are: least squares, least angle regression, random forests, and spline regression. Previous articles (van der Laan and Dudoit (2003); van der Laan et al. (2006); Sinisi et al. (2007)) theoretically validated the use of cross validation to select an optimal learner among many candidate learners. Motivated by this use of cross validation, we propose a new prediction method for creating a weighted combination of many candidate learners to build the super learner. This article proposes a fast algorithm for constructing a super learner in prediction which uses V-fold cross-validation to select weights to combine an initial set of candidate learners. In addition, this paper contains a practical demonstration of the adaptivity of this so called super learner to various true data generating distributions. This approach for construction of a super learner generalizes to any parameter which can be defined as a minimizer of a loss function. PMID- 17910530 TI - A Bayesian approach to estimation and testing in time-course microarray experiments. AB - The objective of the present paper is to develop a truly functional Bayesian method specifically designed for time series microarray data. The method allows one to identify differentially expressed genes in a time-course microarray experiment, to rank them and to estimate their expression profiles. Each gene expression profile is modeled as an expansion over some orthonormal basis, where the coefficients and the number of basis functions are estimated from the data. The proposed procedure deals successfully with various technical difficulties that arise in typical microarray experiments such as a small number of observations, non-uniform sampling intervals and missing or replicated data. The procedure allows one to account for various types of errors and offers a good compromise between nonparametric techniques and techniques based on normality assumptions. In addition, all evaluations are performed using analytic expressions, so the entire procedure requires very small computational effort. The procedure is studied using both simulated and real data, and is compared with competitive recent approaches. Finally, the procedure is applied to a case study of a human breast cancer cell line stimulated with estrogen. We succeeded in finding new significant genes that were not marked in an earlier work on the same dataset. PMID- 17910532 TI - Taking the patient to the classroom: applying theoretical frameworks to simulation in nursing education. AB - Upon completion of their education, nursing students are expected to practice safely and competently. Societal changes and revisions to nursing education have altered the way nursing students learn to competently care for patients. Increasingly, simulation experiences are used to assist students to integrate theoretical knowledge into practice. Reasons for and the variety of simulation activities used in nursing education in light of learning theory are discussed. By combining Benner's nursing skill acquisition theory with Kolb's experiential learning theory, theoretical underpinnings for examining the use of simulations in the context of nursing education are provided. PMID- 17910533 TI - Gcn5- and Elp3-induced histone H3 acetylation regulates hsp70 gene transcription in yeast. AB - The purpose of this study was to elucidate the mechanisms by which histone acetylation participates in transcriptional regulation of hsp70 (heat-shock protein 70) genes SSA3 and SSA4 in yeast. Our results indicated that histone acetylation was required for the transcriptional activation of SSA3 and SSA4. The HATs (histone acetyltransferases) Gcn5 (general control non-derepressible 5) and Elp3 (elongation protein 3) modulated hsp70 gene transcription by affecting the acetylation status of histone H3. Although the two HATs possessed overlapping function regarding the acetylation of histone H3, they affected hsp70 gene transcription in different ways. The recruitment of Gcn5 was Swi/Snf-dependent and was required for HSF (heat-shock factor) binding and affected RNAPII (RNA polymerase II) recruitment, whereas Elp3 exerted its roles mainly through affecting RNAPII elongation. These results provide insights into the effects of Gcn5 and Elp3 in hsp70 gene transcription and underscore the importance of histone acetylation for transcriptional initiation and elongation in hsp genes. PMID- 17910535 TI - Lactococcus lactis as expression host for the biosynthetic incorporation of tryptophan analogues into recombinant proteins. AB - Incorporation of Trp (tryptophan) analogues into a protein may facilitate its structural analysis by spectroscopic techniques. Development of a biological system for the biosynthetic incorpor-ation of such analogues into proteins is of considerable importance. The Gram-negative Escherichia coli is the only prokaryotic expression host regularly used for the incorporation of Trp analogues into recombinant proteins. Here, we present the use of the versatile Gram positive expression host Lactococcus lactis for the incorporation of Trp analogues. The availability of a tightly regulated expression system for this organism, the potential to secrete modified proteins into the growth medium and the construction of the trp-synthetase deletion strain PA1002 of L. lactis rendered this organism potentially an efficient tool for the incorporation of Trp analogues into recombinant proteins. The Trp analogues 7-azatryptophan, 5 fluorotryptophan and 5-hydroxytryptophan were incorporated with efficiencies of >97, >97 and 89% respectively. Interestingly, 5-methylTrp (5-methyltryptophan) could be incorporated with 92% efficiency. Successful biosynthetical incorporation of 5-methylTrp into recombinant proteins has not been reported previously. PMID- 17910534 TI - Biotransformation of p-methoxyphenylacetonitrile into p-methoxyphenylacetic acid by resting cells of Bacillus subtilis. AB - Resting cells of Bacillus subtilis ZJB-063 were used for the direct transformation of MOPAN (p-methoxyphenylacetonitrile) to MOPAA (p methoxyphenylacetic acid), which is an important pharmaceutical intermediate. The B. subtilis ZJB-063 culture conditions for the production of nitrilase and the reaction conditions for this nitrilase-mediated conversion were optimized. The maximum production of nitrilase was achieved when glucose and a combination of ammonium sulfate and yeast powder were added as carbon and nitrogen sources respectively. Previously reported inducers were found to be unnecessary for the production of nitrilase from B. subtilis ZJB-063, which indicated that this nitrilase appeared to be constitutive. However, when epsilon-caprolactam (6 hexanolactam) was added as the inducer, B. subtilis ZJB-063 exhibited nitrile hydratase and amidase activity. The maximum conversion of MOPAN into MOPAA (specific activity 17.03 units.g(-1)(DCW); DCW is dry cell weight) was observed in a solution containing 50 mM phosphate buffer (pH 7.0), 10 mM MOPAN, 2.7 mg DCW.ml(-1) wet resting cells and 5% (v/v) DMSO for 4 h at 32 degrees C. MOPAN (10 mM) was completely converted into MOPAA (9.65 mM) in 5 h in shake flasks without the formation of p-methoxyphenylacetamide. The small deviation of MOPAA (9.65 mM) from the theoretical amount (10 mM) may be due to partial consumption of the products by B. subtilis ZJB-063. Both MOPAN and MOPAA inhibited the hydrolysis at concentrations above 15 mM. Scale up of the reaction to 200 ml in a bubble bioreactor shortened the reaction time compared with the reactions performed in shake flasks. PMID- 17910536 TI - Adults with mild intellectual disabilities: can their reading comprehension ability be improved? AB - BACKGROUND: Adults with a mild intellectual disability (ID) often show poor decoding and reading comprehension skills. The goal of this study was to investigate the effects of teaching text comprehension strategies to these adults. Specific research goals were to determine (1) the effects of two instruction conditions, i.e. strategy instruction to individuals and strategy instruction in small groups in a reciprocal teaching context; (2) intervention programme effects on specific strategy tests (so-called direct effects), and possible differences between strategies; (3) (long-term) transfer effects of the programme on general reading comprehension ability; and (4) the regression of general text comprehension by the variables of technical reading, IQ, reading comprehension of sentences (RCS), and pretest and posttest scores on the strategies taught. METHODS: In total, 38 adults (age range 20-72 years; mean age of 36 years) with ID participated in the study. IQs ranged from 45 to 69 with a mean IQ of 58. The intervention programme involved 15 weekly lessons of 1 h each, taught during 3 months. Blocks of lessons included each of Brown and Palincsar's strategies of summarizing, questioning, clarifying and predicting, as participants read and studied narrative and expository texts. RESULTS: Results indicated no significant difference between group and individual instruction conditions. Second, direct programme effects - as determined by posttest-pretest contrasts for strategy tests - were substantial, except for the questioning strategy. Third, even more substantial was the transfer effect to general text comprehension. Moreover, the results on this test were well maintained at a follow-up test. Finally, the variance of general reading comprehension ability was best explained by the test of RCS, and only moderately by the strategies trained. CONCLUSION: The presently used intervention programme provides a good starting point for adults with ID to become better readers. PMID- 17910537 TI - Self-determination, social abilities and the quality of life of people with intellectual disability. AB - BACKGROUND: The international literature has documented that self-determination is impacted by environmental factors, including living or work settings; and by intraindividual factors, including intelligence level, age, gender, social skills and adaptive behaviour. In addition, self-determination has been correlated with improved quality of life (QoL). This study sought to contribute to the growing literature base in this area by examining the relationship among and between personal characteristics, self-determination, social abilities and the environmental living situations of people with intellectual disabilities (ID). METHODS: The study involved 141 people with ID residing in Italy. Healthcare professionals and social workers who had known participants for at least 1 year completed measures of self-determination, QoL and social skills. Analysis of variance was conducted to verify whether different levels of intellectual impairment were associated with different degrees of the dependent variables. The Pearson product-moment correlation was used to examine any relationships among dependent variables and IQ scores. Finally, discriminant function analysis was used to examine the degree to which IQ score, age, self-determination and social abilities predicted membership in groups that were formed based on living arrangement, and on QoL status (high vs. low). RESULTS: The anova determined, as expected, that participants with more severe ID showed the lowest levels of self determination, QoL and social abilities. Discriminant function analysis showed that (a) individuals attending day centres were distinguished from those living in institutions in that they were younger and showed greater autonomy of choice and self-determination in their daily activities; (b) basic social skills and IQ score predicted membership in the high or low QoL groups; and (c) the IQ score predicted membership in the high or low self-determination groups. A manova conducted to examine gender- and age-level differences on self-determination found gender differences; women had higher self-determination scores than men. CONCLUSIONS: These findings contribute to an emerging knowledge base pertaining to the role of intraindividual and environmental factors in self-determination and QoL. In general, the study replicated findings pertaining to the relative contribution of intelligence to self-determination and QoL, added information about the potential contribution of social abilities, and pointed to the potentially important role of opportunities to make choices as a particularly important aspect of becoming more self-determined, at least in the context of residential settings. PMID- 17910538 TI - Poverty, socio-economic position, social capital and the health of children and adolescents with intellectual disabilities in Britain: a replication. AB - Background When compared with their nonintellectually disabled peers, people with intellectual disabilities (IDs) have poorer health and are more likely to be exposed to poverty during childhood. Given that exposure to child poverty has been linked to poorer health outcomes, we attempted to estimate the extent to which the health inequalities faced by children and adolescents with IDs may be accounted for by their more disadvantaged socio-economic position. Methods Secondary analysis of data on a nationally representative sample of 12 160 British children aged under 17 years extracted from the Department of Work and Pensions' Families and Children Study. Results After controlling for age and sex, children with IDs were significantly more likely (corrected odds ratio = 2.49) to be reported to have less than good health than their nonintellectually disabled peers. However, 31% of the elevated risk for poorer health was accounted for by between-group differences in socio-economic position and social capital. Conclusions A socially and statistically significant proportion of the increased risk of poorer health among children and adolescents with IDs may be attributed to their increased risk of socio-economic disadvantage. PMID- 17910539 TI - Quality of life of adults with intellectual disabilities who live with families in Taiwan. AB - BACKGROUND: Little research has been conducted about the quality of life (QOL) of people with intellectual disabilities (ID) in Taiwan, particularly their subjective QOL. This study examined the personal perceptions of these individuals as measured on internationally recognized core QOL domains and indicators. METHODS: A census interview survey was conducted in Hsin-Chu City in Taiwan; 233 adults aged over 16 years with mild ID and living with their families participated in the study. Data were collected using the Cross-Cultural QOL Indicators (CCQOLI) together with socio-demographic data that included 'activities of daily living' and 'instrumental activities of daily living' (IADL). The CCQOLI were based on the three most commonly reported indicators of each of the eight QOL domains: emotional well-being, interpersonal relations, material well-being, personal development, physical well-being, self determination, social inclusion and rights. Each indicator has two sets of questions related to the indicator's 'importance' and 'use'. These are answered by the respondent using a 4-point Likert scale. RESULTS: The importance and use of the QOL indicators were evaluated positively by the respondents. The adults' individual characteristics, namely IADL and educational level, were significant predictors for the 'importance' while the adults' perceptions of 'use' for overall QOL were significantly affected by his/her socio-economic data, that is, residence location and father's educational level. CONCLUSIONS: The present study addressed the issue of self-reported QOL in people with ID in Taiwanese society, becoming a possible benchmark for similar measurements carried out by disability movements there. These results contribute to current advocacy efforts towards creating a supportive environment for people with ID. PMID- 17910540 TI - Use of the Interact Short Form as a tool to evaluate emotion of people with profound intellectual disabilities. AB - BACKGROUND: One of the essential purposes of intervention programmes for people with profound intellectual disabilities (ID) is to enhance the desirable mood and behaviour and decrease the undesirable ones through stabilizing their emotion. There is lack of validated instrument to offer a comprehensive measure that covers the mood and behaviour, both desirable and undesirable, appropriate for people with profound ID. METHOD: This study aimed to examine the validity and reliability of the Interact Short Form for evaluating the mood and behaviour of people with profound ID, and at the same time, review their emotional profile using the Interact Short Form. Both content validity using expert panel review and construct validity by means of factor analysis were investigated. A total of 75 people with profound ID were recruited. Inter-rater reliability was tested. The results of the Interact Short Form were described to reflect the emotional profile of this group of participants. RESULTS: Using the results of expert panel review and those from factor analysis, we found three subscales representing the mood and behaviour of people with profound ID. They were: 'emotional expression', 'interests towards tasks' and 'behaviours to environment'. All three subscales were found to be internally consistent (alpha = 0.71-0.88). The Interact Short Form- People with profound ID version also showed good inter-rater reliability (mean = 0.72). The results of the Interact Short Form showed that this group of participants had fairly stable emotion under the structured setting and activities in the residential institutions where data were collected. CONCLUSIONS: The Interact Short Form- People with profound ID version serves as a helpful tool for both clinical and research use in assessing the mood and behaviour of people with profound ID in a simple, comprehensive and systematic way. PMID- 17910541 TI - Validation of the Subjective and Objective Family Burden Interview (SOFBI/ECFOS) in primary caregivers to adults with intellectual disabilities living in the community. AB - BACKGROUND: There is little information on the psychometric properties of instruments for assessing family care burden in adults with intellectual disabilities (ID). The aim of this study is therefore to analyse the usefulness of the 'Subjective and Objective Family Burden Interview' (SOFBI) in the assessment of principal caregivers in Spain. METHODS: The SOFBI was administered to 166 principal caregivers of adults with ID in a vocational centre. The psychometric analysis included: internal consistency, inter-rater and test-retest reliability, construct validity, convergent validity with the World Health Organization's Disability Assessment Schedule II, and feasibility. RESULTS: The Cronbach's alpha was 0.88 for the overall interview and always above 0.7 in the quantitative subdomains. The Kappa coefficients for test-retest were between 0.5 and 0.8, whereas inter-rater agreement was nearly perfect. Maximum-likelihood factor analysis showed four well-defined factors, which fitted the previously designed domains. Feasibility was also good. CONCLUSIONS: The SOFBI is a multi domain, modular instrument which is feasible, reliable and valid for measuring the burden of family caregivers to adults with ID living in the community. PMID- 17910542 TI - Cognitive-behavioural treatment for men with intellectual disabilities and sexually abusive behaviour: a pilot study. AB - BACKGROUND: Cognitive-behaviour therapy (CBT) seems to be becoming the treatment of choice for non-disabled sex offenders. Nevertheless, there have been relatively few evaluations of such treatment for men with intellectual disabilities (ID) and sexually abusive behaviour. METHOD: A pilot study providing CBT for two groups of men with ID is described. Measures of change in sexual knowledge, victim empathy and cognitive distortions were collected, together with a log of further sexually abusive behaviour. RESULTS: Fifteen men were offered treatment but some dropped out and some declined to take part in the research. The results for the eight men who consented to the research and completed treatment showed significant positive changes in sexual knowledge and victim empathy (two men completed both groups, making 10 sets of data in all). Cognitive distortions showed significant change on only one of the two measures. Some men showed further sexually abusive behaviour either during or after the treatment group (all had been previously diagnosed as on the autistic spectrum). CONCLUSION: There is a need for a larger multi-site trial of treatment with a broad set of measures and the ability to analyse who benefits from such treatments and who does not. PMID- 17910544 TI - Introduction to the assessing care of vulnerable elders-3 quality indicator measurement set. AB - OBJECTIVES: To update and increase the comprehensiveness of the Assessing Care of Vulnerable Elders (ACOVE) set of process-of-care quality indicators (QIs) for the medical care provided to vulnerable elders and to keep up with the constantly changing medical literature, the QIs were revised and expanded. DESIGN: The ACOVE Clinical Committee expanded the number of measured conditions to 26 in the revised (ACOVE-3) set. For each condition, a content expert created potential QIs and, based on systematic reviews, developed a peer-reviewed monograph detailing each QI and its supporting evidence. Using these literature reviews, multidisciplinary panels of clinical experts participated in two rounds of anonymous ratings and a face-to-face group discussion to evaluate whether the QIs were valid measures of quality of care using a process that is an explicit combination of scientific evidence and professional consensus. The Clinical Committee evaluated the coherence of the complete set of QIs that the expert panels rated as valid. RESULTS: ACOVE-3 contains 392 QIs covering 14 different types of care processes (e.g., taking a medical history, performing a physical examination) and all four domains of care: screening and prevention (31% of QIs), diagnosis (20%), treatment (35%), and follow-up and continuity (14%). All QIs also apply to community-dwelling patients aged 75 and older. CONCLUSION: ACOVE-3 contains a set of QIs to comprehensively measure the care provided to vulnerable older persons at the level of the health system, health plan, or medical group. These QIs can be applied to identify areas of care in need of improvement and can form the basis of interventions to improve care. PMID- 17910543 TI - Hearing aids: expectations and satisfaction of people with an intellectual disability, a descriptive pilot study. AB - BACKGROUND: In spite of an increased risk of hearing impairment in persons with an intellectual disability (ID), rehabilitation with hearing aids often fails. We performed a descriptive pilot study with the following study questions: 1 Do comparable elements as in the general population contribute to expectations of and satisfaction with hearing aids in adults with mild or moderate IDs? 2 To what extent do adults with an ID depend on carers in use and maintenance of hearing aids? METHOD: STUDY POPULATION: 16 adults with a mild or moderate ID and a recent diagnosis of hearing impairment. Method: information by means of specially designed booklets; semi-structured interviews prior to hearing aid fitting and 6 months afterwards. ANALYSIS: descriptive. RESULTS: In total, 14/16 participants were able to give reliable answers. Most were aware of their hearing loss and familiar with reasons for hearing aids. A minority expressed positive expectations. Some expressed explicit wishes on the looks of hearing aids. All satisfaction domains as described for the general population could be recognized. Most participants were partially or totally dependent on carers in use and maintenance of hearing aids. CONCLUSIONS: Adults with ID may have explicit ideas and wishes about hearing aids and, if specifically asked, are capable of expressing these. Given information should be checked and repeated. In satisfaction with hearing aids, comparable elements may play a role as in the general population: benefit, cosmetics, sound quality/acoustics, comfort/ease of use, and service delivery. These findings, however, are from a small-scale study. Additional research is necessary to find out whether they are applicable more generally. PMID- 17910545 TI - Quality indicators for benign prostatic hyperplasia in vulnerable elders. PMID- 17910546 TI - Quality indicators for the care of breast cancer in vulnerable elders. PMID- 17910547 TI - Quality indicators for the care of chronic obstructive pulmonary disease in vulnerable elders. PMID- 17910548 TI - Quality indicators for the care of colorectal cancer in vulnerable elders. PMID- 17910549 TI - Quality indicators for continuity and coordination of care in vulnerable elders. PMID- 17910550 TI - Quality indicators for the care of dementia in vulnerable elders. PMID- 17910551 TI - Quality indicators for the care of depression in vulnerable elders. PMID- 17910552 TI - Quality indicators for the care of diabetes mellitus in vulnerable elders. PMID- 17910553 TI - Quality indicators for palliative and end-of-life care in vulnerable elders. PMID- 17910554 TI - Quality indicators for falls and mobility problems in vulnerable elders. PMID- 17910555 TI - Quality indicators for the care of hearing loss in vulnerable elders. PMID- 17910556 TI - Quality indicators for the care of heart failure in vulnerable elders. PMID- 17910557 TI - Quality indicators for hospitalization and surgery in vulnerable elders. PMID- 17910558 TI - Quality indicators for the care of hypertension in vulnerable elders. PMID- 17910559 TI - Quality indicators for the care of ischemic heart disease in vulnerable elders. PMID- 17910560 TI - Quality indicators for medication use in vulnerable elders. PMID- 17910561 TI - Quality indicators for the care of osteoarthritis in vulnerable elders. PMID- 17910562 TI - Quality indicators for the care of osteoporosis in vulnerable elders. PMID- 17910564 TI - Quality indicators for the care of pressure ulcers in vulnerable elders. PMID- 17910563 TI - Quality indicators for pain management in vulnerable elders. PMID- 17910565 TI - Quality indicators for screening and prevention in vulnerable elders. PMID- 17910566 TI - Quality indicators for the care of sleep disorders in vulnerable elders. PMID- 17910567 TI - Quality indicators for the care of stroke and atrial fibrillation in vulnerable elders. PMID- 17910568 TI - Quality indicators for the care of undernutrition in vulnerable elders. PMID- 17910569 TI - Quality indicators for the screening and care of urinary incontinence in vulnerable elders. PMID- 17910570 TI - Quality indicators for the care of vision impairment in vulnerable elders. PMID- 17910571 TI - Application of assessing care of vulnerable elders-3 quality indicators to patients with advanced dementia and poor prognosis. AB - OBJECTIVES: To use a formal decision-making strategy to reach clinically appropriate, internally consistent decisions on the application of quality indicators (QIs) to vulnerable elders (VEs) with advanced dementia (AD) or poor prognosis (PP). DESIGN: Using a conceptual model that classifies QIs principally by aim and burden of the care process, 12 clinical experts rated whether each Assessing Care of Vulnerable Elders-3 (ACOVE-3) QI should be applied in evaluating quality of care for older persons with AD or PP. QI exclusions were assessed for each of the 26 conditions and by whether these conditions were mainly medical (e.g., diabetes mellitus), geriatric (e.g., falls), or crosscutting processes of care (e.g., pain management). QI exclusions were also identified for older persons who decided against hospitalization or surgery. RESULTS: Of 392 ACOVE-3 QIs, 140 (36%) were excluded for patients with AD and 135 (34%) for patients with PP; 57% of QIs focusing on medical conditions were excluded from patients with AD and 53% from patients with PP, whereas only 20% of QIs for geriatric conditions were excluded from AD and 15% from PP. All QIs with care processes judged to carry a heavy burden were excluded; 86% of moderate burden QIs were excluded from AD and 92% from PP. All QIs aimed at long-term goals were excluded; 83% of intermediate-term goal QIs were excluded from AD and 98% from PP. Individuals holding a preference to forgo hospitalization or surgery would be excluded from 7% of potentially applicable QIs. CONCLUSION: Measurement of quality of care for VEs with AD, PP, and less-aggressive care preferences should include only a subset of the ACOVE-3 QIs, largely those whose burden is light and whose goal is continuity or short-term improvement or prevention. PMID- 17910572 TI - Assessing care of vulnerable elders-3 quality indicators. PMID- 17910573 TI - Proceedings of the 8th Workshop on the Neurobiology of Epilepsy (WONOEP VIII), August 24-26, 2005, Villiers-le-Mahieu, France. PMID- 17910575 TI - Intrahippocampal injection of endothelin-1: a new model of ischemia-induced seizures in immature rats. AB - The goal of this study was to develop a new model of ischemia-induced seizures in immature rats using injection of vasoconstrictor Endothelin-1 (ET-1) into the brain. ET-1 (10, 20, or 40 pmol) was infused into the left dorsal hippocampus of freely moving Wistar rats 12 (P12) and 25 (P25) days old. Animals were then video/EEG-monitored for 100 min and monitoring was repeated 22 h later. Parameters of electrographic seizures (frequency and mean duration) as well as pattern of their behavioral correlates were evaluated. The pattern of behavioral seizures was used to develop model-specific scoring system. Cresyl violet and Fluoro Jade-B-staining were used to evaluate brain damage. Extension of the lesion was correlated with seizure severity. After ET-1-injection, seizures occurred in 83-100% animals of all age-and-dose groups and persisted for 24 h except P12 rats with 10 pmol. There were no differences in average seizure duration (18-40 s) or seizure frequency (3-7 seizures/100 min) among individual dose-groups. Between the 1st and 2nd observation period, total seizure duration decreased in 71% of P12 and 47% of P25 rats. Electrographic seizure activity was most frequently accompanied by clonus, incidence of more severe convulsions (barrel rolling or generalized clonic seizures) increased with dose of ET-1. Morphologic examination did not reveal any dose-related difference in damage severity, hippocampal damage was however more extensive in P12 compared to P25 animals. Seizure severity correlated positively with severity of the damage in both age groups. Our study presents focal injection of ET-1 into the brain as a new and practical model of ischemia-induced seizures in immature rats. PMID- 17910574 TI - Epileptogenesis in the developing brain: what can we learn from animal models? AB - Knowledge of the processes by which epilepsy is generated (epileptogenesis) is incomplete and has been a topic of major research efforts. Animal models can inform us about these processes. We focus on the distinguishing features of epileptogenesis in the developing brain and model prolonged febrile seizures (FS) that are associated with human temporal lobe epilepsy. In the animal model of FS, epileptogenesis occurs in approximately 35% of rats. Unlike the majority of acquired epileptogeneses in adults, this process early in life (in the febrile seizures model as well as in several others) does not require "damage" (cell death). Rather, epileptogenesis early in life involves molecular mechanisms including seizure-evoked, long-lasting alterations of the expression of receptors and ion channels. Whereas transient changes in gene expression programs are common after early-life seizures, enduring effects, such as found after experimental FS, are associated with epileptogenesis. The ability of FS to generate long-lasting molecular changes and epilepsy suggests that mechanisms, including cytokine activation that are intrinsic to FS generation, may play a role also in the epileptogenic consequences of these seizures. PMID- 17910576 TI - Developmental patterns in the regulation of chloride homeostasis and GABA(A) receptor signaling by seizures. AB - GABA(A) receptors have dual functions during development. They depolarize immature neurons but hyperpolarize more mature neurons. This functional switch has been attributed to age-related differences in the relative abundance of cation chloride cotransporters, such as KCC2 and NKCC1, which regulate chloride homeostasis. Certain insults, such as trauma, ischemia, and seizures, if they occur when GABA(A)ergic signaling is hyperpolarizing, such as in the adult brain, can lead to reappearance of the immature, depolarizing synaptic responses to GABA(A) receptor activation. In certain cases, this has been associated with either reduced expression of KCC2 or increase in NKCC1. In epilepsy, the depolarizing effects of GABA(A) receptors have been proposed to be important for the acquisition and/or maintenance of the epileptic state. Using the kainic acid model of status epilepticus, we have studied the effects of repetitive neonatal episodes of status epilepticus on the expression of cation chloride cotransporter KCC2 in the neonatal hippocampus. In contrast to adults, seizures increased KCC2 mRNA expression in the CA3 region of the neonatal hippocampus. The contrasting patterns of regulation of KCC2 by seizures in mature and immature neurons may be one of the age-related factors that protect the neonatal brain against the development of epilepsy. PMID- 17910577 TI - Neurodevelopmental impact of antiepileptic drugs and seizures in the immature brain. AB - Seizure incidence during the neonatal period is higher than any other period in the lifespan, yet we know little about this period in terms of the effect of seizures or of the drugs used in their treatment. The fact that several antiepileptic drugs (AEDs) induce pronounced apoptotic neuronal death in specific regions of the immature brain prompts a search for AEDs that may be devoid of this action. Furthermore, there is a clear need to find out if a history of seizures alters the proapoptotic action of the AEDs. Our studies are aimed at both of these issues. Phenytoin, valproate, phenobarbital, and MK801 each induced substantial regionally specific cell death, whereas levetiracetam even in high doses (up to 1,500 mg/kg) did not have this action. In view of our previously findings of neuroprotective actions of repeated seizures in the adult brain, we also examined repeated seizures for a possible antiapoptotic action in the infant rat. Rat pups were preexposed to electroshock seizures (ECS) for 3 days (age 5-7 days) before receiving MK801 on day 7. The effect of ECS, which was consistently a 30% decrease in MK801-induced programmed cell death (PCD), suggests that repeated seizures can exert an antiapoptotic action in the infant brain. In contrast, PCD induced by valproate was not attenuated by ECS preexposure, suggesting that valproate-induced PCD is mechanistically distinct from that induced by MK801 and may not be activity-dependent. Presently, we do not know if this neuroprotective effect of seizures is deleterious or beneficial. If the seizures also enhance the survival of neurons that are destined to undergo naturally occurring PCD, early childhood seizures may have deleterious effects by preventing this necessary component of normal development. While this effect of seizures might be counteracted by AEDs, the fact that several AEDs shift the PCD to the other extreme, and trigger excessive neuronal cell loss, raises concern about whether the drug therapy may be more detrimental than the seizures. In this context, it is encouraging that we have identified at least one AED that is devoid of a proapoptotic action in the infant brain, even in high doses. It is now important to evaluate the long-term consequences of the changes in PCD in infancy by examining behavioral outcomes and seizure susceptibility in the AED- and seizure-exposed neonates when they reach adulthood. PMID- 17910578 TI - Inflammation exacerbates seizure-induced injury in the immature brain. AB - We examined the hypothesis that the introduction of an inflammatory agent would augment status epilepticus (SE)-induced neuronal injury in the developing rat brain in the absence of an increase in body temperature. Postnatal day 7 (P7) and P14 rat pups were injected with an exogenous provocative agent of inflammation, lipopolysaccharide (LPS), 2 h prior to limbic SE induced by either lithium pilocarpine (LiPC) or kainic acid. Core temperature was recorded during the SE and neuronal injury was assessed 24 h later using profile cell counts in defined areas of the hippocampus. While LPS by itself did not produce any discernible cell injury at either age, it exacerbated hippocampal damage induced by seizures. In the LiPC model, this effect was highly selective for the CA1 subfield, and there was no concomitant rise in body temperature. Our findings show that inflammation increases the vulnerability of immature hippocampus to seizure induced neuronal injury and suggest that inflammation might be an important factor aggravating the long-term outcomes of seizures occurring early in life. PMID- 17910579 TI - Voltage depth profiles of high-frequency oscillations after kainic acid-induced status epilepticus. AB - High-frequency oscillations (HFOs) have been described in normal and epileptic brains of animals and humans. These oscillations reflect a short-term integration within neuronal networks and have important functional consequences for normal and pathological processes. We performed a comparative voltage depth profile analysis of normal and pathological HFOs after intrahippocampal kainic acid injection. Sixteen channel recording probes, with 100-200 microm separation between the tips of microelectrodes, were implanted along the CA1-dentate gyrus axis in the anterior hippocampus of adult rats. Guide cannulae were implanted in the CA3 area. After a week of baseline recording kainic acid (KA) (0.2microg/0.2microl) was injected into the CA3 area. Electrical activity continued to be record for the next 3-4 weeks after KA induced status epilepticus. Voltage depth profiles and power spectral analysis of HFOs were performed off-line using DataPac software. Ripple oscillations (80-200 Hz) in the CA1 area and gamma activity (40-80 Hz) in the dentate gyrus remained after status epilepticus. In the group of rats that later developed seizures a new pattern consisting of bursts of population spikes (BPS) occurred. The maximum of amplitude for BPS generated in CA1 was in the pyramidal layer and for those generated in the dentate gyrus was in the granular layer. BPS appeared 2-3 days after status epilepticus and remained for the rest of the experiments. The frequencies of intraburst spikes varied between 80 Hz and 600 Hz. With increasing distance from the area of the burst generation, this activity took on the appearance of HFOs. The occurrence of spontaneous BPS appear to be a primary electrophysiological consequence of status epilepticus when progressive epileptogenesis occurs with maximum of amplitude in the cellular layer. In areas outside of the generator of the BPS, this activity looks more like pathological high-frequency oscillations (pHFO), which were observed in earlier experiments. PMID- 17910580 TI - Pathogenesis and pharmacology of epilepsy in the lithium-pilocarpine model. AB - To try to identify the critical structures during epileptogenesis, we used the lithium-pilocarpine model that reproduces most clinical and neuropathological features of temporal lobe epilepsy (TLE). We used imaging techniques as well as a disease modifying approach and pharmacological strategy. With [14C]-2 deoxyglucose autoradiography, we assessed changes in cerebral glucose utilization. T2-weighted magnetic resonance imaging (MRI, 4.7 T) allowed follow up of structures involved in epileptogenesis. A potential disease-modifying effect was studied using preconditioning with brief seizures (amygdala kindling, maximal electroshocks) and pharmacological strategies including vigabatrin (250 mg/kg), caffeine (0.3 g/L in drinking water), topiramate (10-60 mg/kg), pregabalin (50 mg/kg followed by 10 mg/kg), or RWJ-333369 (10-120 mg/kg). In adult and PN21 rats that became epileptic, entorhinal, and piriform cortices were the initial structures exhibiting significant signal changes on MRI scans, from 6 h after status epilepticus (SE) onset, reflecting neuronal death. In PN21 rats that did not become epileptic, no signal occurred in parahippocampal cortices. In hippocampus, MRI signal change appeared 36-48 h after SE, and progressively worsened to sclerosis. During the latent and chronic phases, the metabolic level in the hilus of adult and PN21 epileptic rats was normal although neuronal loss reached 60-75%. Protection limited to CA1 and/or CA3 (caffeine, topiramate, vigabatrin, amygdala kindling) did not affect the latency to spontaneous seizures. Protection limited to the entorhinal and piriform cortices (pregabalin) delayed epileptogenesis. The combined protection of Ammon's horn and parahippocampal cortices (RWJ-333369) prolonged the latency before the onset of seizures in a dose-dependent manner or, in some cases, prevented the epilepsy. The entorhinal and piriform cortices are critically involved in the early phase of the epileptogenesis while the hilus may initiate and/or maintain epileptic seizures. Pharmacological protection of the basal cortices is necessary for a beneficial disease-modifying effect but this must be combined with protection of the hippocampus to prevent epileptogenesis in this model of TLE. PMID- 17910581 TI - Chemoconvulsant-induced seizure susceptibility: toward a common genetic basis? AB - Despite the efforts employed, understanding the genetic architecture underlying epilepsy remains difficult. To reach this aim, convulsive epilepsies are classically modeled in mice, where genetic studies are less constricting than in humans. Pharmacogenetic approaches are one major source of investigation where kainic acid, pentylenetetrazol, and the ss-carboline family represent compounds that are used extensively. Several quantitative trait loci (QTLs) influencing the convulsant effects of these drugs have been mapped using either recombinant inbred strains (RIS) or segregating F2 populations (or both). In our laboratory, we have recently mapped two QTLs for methyl 6, 7-dimethoxy-4-ethyl-ss-carboline-3 carboxylate (DMCM), and seizure response using an F2 method. One is located on the distal part of Chromosome 1, a region implicated in a number of other studies. Here, we address the general importance of this chromosomal fragment for influencing seizure susceptibility. PMID- 17910582 TI - Dynamic changes of proteases and protease inhibitors revealed by microarray analysis in CA3 and entorhinal cortex during epileptogenesis in the rat. AB - We investigated expression of genes involved in the proteolytic process during epileptogenesis in a rat model of temporal lobe epilepsy (TLE). In a previous microarray study we found prominent activation of this process, which reached highest expression during the acute and latent phase (1 week after SE) in CA3 and entorhinal cortex (EC). Detailed analysis shows differences in dynamics of the changes of several protease genes such as cathepsins, caspases, matrix metalloproteinases, and plasminogen activators. Most genes were acutely upregulated while others were mainly activated during the latent phase. Interestingly several proteolytic genes were still elevated in the chronic epileptic phase. Various protease inhibitors followed a similar time course. The identification of changes in the activation of genes involved in proteolysis at critical phases during epileptogenesis could point to potential time specific targets for intervention. The fact that several proteolytic genes were still activated in the chronic epileptic phase makes them interesting candidates to modify and slow down seizure progression. PMID- 17910583 TI - Alterations of phosphatidylinositol 3-kinase pathway components in epilepsy associated glioneuronal lesions. AB - Low-grade glioneuronal lesions involving tumors such as gangliogliomas and focal cortical dysplasias (FCD) predispose individuals to pharmacoresistant epilepsy. A frequent variant of FCD is composed of dysplastic cytomegalic neurons and Taylor type balloon cells (FCD(IIb)). Those are similar to cellular elements, which are present in cortical tubers in the autosomal dominant inherited tuberous sclerosis complex (TSC). This phacomatosis is caused by mutations in the TSC1 or TSC2 genes. Recent data have indicated accumulation of distinct allelic variants of TSC1 also in FCD(IIb). TSC1 represents a key factor in the phosphatidylinositol 3 kinase (PI3K) pathway. A variety of alterations in the PI3K-pathway have been recently reported in epilepsy-associated glioneuronal malformations. Here, we discuss pathogenetic similarities and differences between cortical dysplasias as well epilepsy-associated glioneuronal tumors and TSC-associated cortical tubers with a focus on PI3K-pathway components including ezrin, radixin and moesin (ERM), which represent downstream effectors involved in cytoskeleton-membrane interference. No evidence has been found for mutational events of ERM genes to play a major pathogenetic role in epilepsy-associated glioneuronal malformations. In contrast, aberrant expression of ERM proteins in FCDs and gangliogliomas was observed. These alterations may relate to compromised interactions of dysplastic cellular components in epilepsy-associated glioneuronal lesions and be involved in aberrant PI3K-pathway signaling in epilepsy-associated malformations. However, the underlying cause of PI3K-pathway activation and the functional relationship of PI3K-pathway activity to generation of seizures in epilepsy-associated glioneuronal lesions will need to be determined in the future. PMID- 17910584 TI - A hypothesis regarding the pathogenesis and epileptogenesis of pediatric cortical dysplasia and hemimegalencephaly based on MRI cerebral volumes and NeuN cortical cell densities. AB - This study compared MRI cerebral volumes and Neuronal-Nuclei (NeuN) cell densities in pediatric epilepsy surgery patients with cortical dysplasia (CD; n = 25) and hemimegalencephaly (HME; n = 14). Our purpose was to deduce possible mechanisms of pathogenesis and epileptogenesis based on an understanding of normal developmental corticoneurogenesis. We used MRI to measured cerebral hemisphere volumes, and NeuN staining to determine grey and white matter cell densities and cell sizes in the molecular layer, grey, and white matter. CD and HME surgical cases were compared with autopsy or non-CD cases (n = 20). Total MRI brain volumes were similar between non-CD, CD, and HME cases. However, in HME patients, the affected cerebral hemisphere was larger and the nonaffected side smaller than non-CD cases. Compared with autopsy cases, NeuN cell densities and cell sizes in CD and HME patients were increased in the molecular layer, upper grey matter, and white matter. In CD and HME cases, total cerebral hemisphere volumes were normal in size and there were more cortical neurons in upper layers than expected. The increase in cortical neuronal densities is consistent with the hypothesis that CD and HME pathogenesis involves increased neurogenesis in the late (not early) phases of cortical formation. In addition, more neurons in the molecular layer and white matter supports the concept that CD and HME pathogenesis also involves incomplete programmed cell death in the remnant cells occupying the preplate and subplate regions. Based on our anatomical and previous electrophysiological findings, we propose that in CD and HME seizure generation is the consequence of incomplete cerebral development with abnormal interactions between immature and mature cells and cellular networks. PMID- 17910585 TI - Immature neurons and GABA networks may contribute to epileptogenesis in pediatric cortical dysplasia. AB - Cortical dysplasia (CD), a frequent pathological substrate of pediatric epilepsy surgery patients, has a number of similarities with immature cortex, such as reduced Mg2+ sensitivity of N-methyl-D-aspartate (NMDA) receptors and the persistence of subplate-like neurons and undifferentiated cells. Because gamma aminobutyric acid (GABA) is the main neurotransmitter in early cortical development, we hypothesized increased GABA receptor-mediated synaptic function in CD tissue. Infrared videomicroscopy and whole-cell patch clamp recordings were used to characterize the morphology and electrophysiological properties of immature and normal-appearing neurons in slices from cortical tissue samples resected for the treatment of pharmacoresistant epilepsy in children (0.2-14 years). In addition, we examined spontaneous and evoked synaptic activity, as well as responses to exogenous GABA application. We demonstrate both the presence of immature pyramidal neurons and networks in young CD tissue and the predominance of GABA synaptic activity. In addition, spontaneous GABA depolarizations frequently induced action potentials, supporting a potential excitatory role of GABA in CD. Evoked synaptic responses mediated by GABA were also prominent, and bath application of 4-aminopyridine induced rhythmic depolarizations that were blocked by bicuculline. Finally, responses to exogenous application of GABA had depolarized reversal potentials in severe compared to mild and non-CD cases. The present data support the hypothesis that CD shares features of immature cortex, with predominant and potentially excitatory GABA(A) receptor-mediated neurotransmission. These results could partially explain the increased excitability of the cortical network in pediatric CD. PMID- 17910586 TI - Activity-dependent gene expression correlates with interictal spiking in human neocortical epilepsy. AB - Interictal spikes are hallmarks of epileptic neocortex that are used commonly in both EEG and electrocorticography (ECoG) to localize epileptic brain regions. Despite their prevalence, the exact relationship between interictal spiking and the molecular pathways that drive the production and propagation of seizures is not known. We have recently identified a common group of genes induced in human epileptic foci, including EGR1, EGR2, c-fos, and MKP-3. We found that the expression levels of these genes correlate precisely with the frequency of interictal activity and can thus serve as markers of epileptic activity. Here, we explore this further by comparing the expression of these genes within human epileptic neocortex to both ictal and specific electrical parameters of interictal spiking from subdural recordings prior to surgical resection in order to determine the electrical properties of the human neocortex that correlate best to the expression of these genes. Seizure frequency as well as quantitative electrophysiological parameters of interictal spikes including frequency, amplitude, duration, and area were calculated at each electrode channel and compared to quantitative real-time RT-PCR measurements of four activity-dependent genes (c-fos, EGR1, EGR2, and MKP-3) in the underlying neocortical tissue. Local neocortical regions of seizure onset had consistently higher spike firing frequencies and higher spike amplitudes compared to nearby "control" cortex. In contrast, spike duration was not significantly different between these two areas. There was no relationship observed between seizure frequency and the expression levels of activity-dependent genes for the patients examined in this study. However, within each patient, there were highly significant correlations between the expression of three of these genes (c-fos, EGR1, EGR2) and the frequency, amplitude, and total area of the interictal spikes at individual electrodes. We conclude that interictal spiking is closely associated with the expression of a group of activity-dependent transcription factors in neocortical human epilepsy. Since there was little correlation between gene expression and seizure frequency, our results suggest that interictal spiking is a stronger driving force behind these activity-dependent gene changes and may thus participate in the development and maintenance of the abnormal neuronal hyperactivity seen in human epileptic neocortex. PMID- 17910587 TI - Timing of the developmental switch in GABA(A) mediated signaling from excitation to inhibition in CA3 rat hippocampus using gramicidin perforated patch and extracellular recordings. AB - The timing of the developmental switch in the GABA(A) mediated responses from excitatory to inhibitory was studied in Wistar rat CA3 hippocampal pyramidal cells using gramicidin perforated patch-clamp and extracellular recordings. Gramicidin perforated patch recordings revealed a gradual developmental shift in the reversal potential of synaptic and isoguvacine-induced GABA(A) mediated responses from -55 +/- 4 mV at postnatal days P0-2 to -74 +/- 3 mV at P13-15 with a midpoint of disappearance of the excitatory effects of GABA at around P8. Extracellular recordings in CA3 pyramidal cell layer revealed that the effect of isoguvacine on multiple unit activity (MUA) switched from an increase to a decrease at around P10. The effect of synaptic GABA(A) mediated responses on MUA switched from an increase to a decrease at around P8. It is concluded that the developmental switch in the action of GABA via GABA(A) receptors from excitatory to inhibitory occurs in Wistar rat CA3 pyramidal cells at around P8-10, an age that coincides with the transition from immature to mature hippocampal rhythms. We propose that excitatory GABA contributes to enhanced excitability and ictogenesis in the neonatal rat hippocampus. PMID- 17910588 TI - The involvement of the substantia nigra pars reticulata in hypoglycemic seizures. AB - Neurological complications of hypoglycemia often include seizures and fasting is a predisposing factor for seizures to occur. The mechanisms involved are unknown. In rats, insulin administration induces hypoglycemia, which may lead to generalized seizures with barrel rotations as a hallmark. Here we compared the incidence of barrel rotations in fasted and nonfasted rats. Further, we investigated the role of the substantia nigra pars reticulata (SNR) in control of barrel rotations using localized bilateral microinfusions of GABA(A) or GABA(B) receptor agonists (muscimol or baclofen, respectively) or an N-methyl-D-aspartate (NMDA) receptor antagonist (AP7). The incidence of barrel rotations was significantly higher in fasted compared to nonfasted rats. SNR infusions of muscimol were ineffective, while both baclofen and AP7 significantly decreased occurrence of barrel rotations. These data suggest that during hypoglycemia, the SNR seizure controlling system has different properties than in seizure models not involving a metabolic stress. PMID- 17910589 TI - GABA(A) receptor internalization during seizures. AB - A rapid modification in the postsynaptic gamma-aminobutyric acid (GABA(A)) receptor population occurs during the prolonged seizures of status epilepticus (SE). This rapid modification contributes to a reduction in GABA-mediated inhibition and the development of benzodiazepine pharmacoresistance. Previous hypotheses to explain the modification have included an alteration in the structural composition or posttranslational modification of the receptors. In a cultured hippocampal neuron model, we found that there was differential subcellular distribution of GABA(A) receptor subunits and that the constitutive internalization of GABA(A) receptors containing a beta2/3 subunit was rapid and activity-dependent. Based on this finding, we posit that an activity-dependent increase in the rate of internalization of synaptic GABA(A) receptors during SE contributes to the reduction in inhibitory transmission and the development of benzodiazepine pharmacoresistance. PMID- 17910590 TI - Neurodevelopment in seizure-prone and seizure-resistant rat strains: recognizing conflicts in management. AB - Cytoarchitectural alterations during central nervous system (CNS) development are believed to underlie aberrations in brain morphology that lead to epilepsy. We have recently reported marked reductions in hippocampal and white matter volumes along with relative ventriculomegaly in a rat strain bred to be seizure-prone (FAST) compared to a strain bred to be seizure-resistant (SLOW) (Gilby et al., 2002, American Epilepsy Society 56th Annual Meeting). This study was designed to investigate deviations in gene expression during late-phase embryogenesis within the brains of FAST and SLOW rats. In this way, we hoped to identify molecular mechanisms operating differentially during neurodevelopment that might ultimately create the observed differences in brain morphology and/or seizure susceptibility. Using Superarray technology, we compared the expression level of 112 genes, known to play a role in neurodevelopment, within whole brains of embryonic day 21 (E21) FAST and SLOW rats. Results revealed that while most genes investigated showed near equivalent expression levels, both Apolipoprotein E (APOE) and the beta2 subunit of the voltage-gated sodium channel (SCN2beta) were significantly underexpressed in brains of the seizure-prone embryos. Currently, these transcripts have no known interactions during embryogenesis; however, they have both been independently linked to seizure disposition and/or neurodevelopmental aberrations leading to epilepsy. Thus, alterations in the timing and/or degree of expression for APOE and SCN2beta may be important to developmental cascades that ultimately give rise to the differing brain morphologies, behaviors, and/or seizure vulnerabilities that characterize these strains. PMID- 17910591 TI - Cell cycle reentry and cell proliferation as candidates for the seizure predispositions in the hippocampus of EL mouse brain. AB - We have recently found that there was DNA fragmentation without cell loss in the hippocampus in EL mice, an epileptic mutant. Neurotrophic factors are also expressed at high levels during the early developmental stages. In the present study, we used EL mice to examine how altered cyclin and the corresponding cyclin dependent kinase (CDK) family are related to cell proliferation during development and during epileptogenesis. Developmental changes of cyclin family and corresponding CDK family (cyclin D/CDK-4, cyclin E/CDK-2, cyclin A/CDK-2, cyclin A/CDK-1, cyclin B/CDK-1) were examined by Western blotting in the hippocampus of EL mice and in nonepileptic control animals (DDY mice). In addition, we attempted to quantify cell proliferation during this period. The developmental changes in cell proliferation were determined by using systemic injections of Bromo-deoxyUridine (BrdU) to label dividing cells. As compared with the control DDY mice, EL mice show an upregulation of cell cycle specific Cyclins/CDKs during early developmental stages suggesting that reentry into the cell cycle is enhanced prior to the onset of seizure activity, possibly due to the abundance of neurotrophic factors. These results show that Cyclins/CDKs are activated during early stages of development in an epileptic animal, before the mouse exhibits seizures. These results suggest that reentry of cells into the cell cycle, with consequent cell proliferation in the hippocampus, contribute to the seizure predispositions of EL mice. PMID- 17910592 TI - Cholinergic dysfunction in temporal lobe epilepsy. AB - The entorhinal cortex-hippocampus complex is believed to be the site of origin of seizure activity in the majority of patients with temporal lobe epilepsy (TLE). Both these regions are enriched with cholinergic innervation, which plays a key role in the normal control of neuronal excitability and in higher cognitive processes. In TLE, anatomical and functional changes occur in all cellular components of the local neural circuit. Thus, while it is not surprising that cholinergic functions are altered in the epileptic temporal lobe, the exact nature and role of these changes in the pathogenesis of the disease are not known. In this report, we summarize the scientific background and experimental data supporting a "cholinergic hypothesis of TLE." We conclude that while the exact role of cholinergic dysfunction in TLE is not known, there is a firm basis for suggesting that changes in the expression of key cholinergic proteins-and the associated cholinergic dysfunction-are key factors in the basic mechanisms underlying TLE. PMID- 17910593 TI - Hyperexcitability of the CA1 hippocampal region during epileptogenesis. AB - Temporal Lobe Epilepsy (TLE) is often preceded by a latent (seizure-free) period during which complex network reorganizations occur. In experimental epilepsy, network hyperexcitability is already present during the latent period, suggesting a modification of information processing. The purpose of this study was to assess the input/output relationship in the hippocampal CA1 region during epileptogenesis. Field recordings in strata pyramidale and radiatum were used to measure the output of CA1 pyramidal cells as a function of the synaptic inputs they receive following the stimulation of Shaffer collaterals in slices obtained from sham and pilocarpine-treated animals during the latent and chronic periods. We show that there is a transient increase of the input and output field responses during the latent period as compared to sham and epileptic animals. The coupling between excitatory inputs and cell firing was also increased during the latent period. This increase persisted in epileptic animals, although to a lesser extent. We also confirm that paired-pulse facilitation occurs before the chronic phase. The present data further support the view that hyperexcitability is present at an early stage of epileptogenesis. Network output is more facilitated during the latent than during the chronic period. Hyperexcitability may participate to epileptogenesis, but it is not sufficient in itself to produce seizures. PMID- 17910594 TI - ABC transporters during epilepsy and mechanisms underlying multidrug resistance in refractory epilepsy. AB - It is estimated 20-25% of the epileptic patients fails to achieve good control with the different antiepileptic drugs (AEDs) treatments, developing refractory epilepsy (RE). Discovered first in cancer, the activity of P-glycoprotein (P-gp) and others ABC transporters as multidrug-resistance-associated proteins (MRPs) and breast cancer resistant protein (BCRP) are directly related with the refractoriness. We have observed the overexpression of these all transporters in the brain of patients with RE, and according with other authors, all these data suggests an active drug efflux from brain. Both constitutive and seizure induced brain P-gp overexpression was also suggested. As confirmation of these clinical evidences, different models of experimental epilepsy have demonstrated P-gp overexpression on blood brain barrier (BBB) and brain parenchyma cells, as astrocytes and neurons. In our model, early P-pg detection in vessel-related cells and later additional P-gp detection in neurons, correlated with the gradual loss of protective effect of phenytoin. The progressive neuronal P-gp expression, depending on intensity and time-constancy of seizure-injury, was in agreement with the development of "P-gp-positive seizure-axis" proposed by Kwan & Brodie, who also showed that the development of RE directly correlated with the number and frequency of seizures before initiation of drug therapy. P-gp expression in excretory organs suggests that P-gp have a central role in drug elimination. Persistent low levels of AEDs in plasma and P-gp brain overexpression in several RE pediatric patients were reported. We also observed in adult RE patients, an increased liver clearance of 99mTc-hexakis-2-methoxyisobutylisonitrile (99mTc MIBI) (a P-gp substrate), and the surgically treated cases showed P-gp brain overexpression. These results suggest the systemic hyperactivity of P-gp in RE patients, including brain P-gp over-expression should be suspected when persistent subtherapeutic levels of AEDs in plasma are detected. P-gp neuronal expression described in both clinical and experimental reports indicates that additional mechanisms could be operative from seizure-affected P-gp-positive neurons, due to AEDs targets are expressed at membrane level. An alternative mechanism was demonstrated in P-gp-expressed cells that exhibit lower membrane potential (Deltapsi(0)=-10 to -20) compared to normal physiological Deltapsi(0) of -60 mV. Under this situation and irrespective to the P-gp pharmacoresistant property or type of drug treatment selected, P-gp-expressed neurons could increase their sensitivity to new seizures perhaps as an epileptogenic mechanism. The understanding of properties of these ABC transporters can offer new tools for better selection of more effective preventive or therapeutic strategies and avoid the invasive surgical treatments for RE. PMID- 17910595 TI - The effect of generalized absence seizures on the progression of kindling in the rat. AB - The involvement of the thalamus in limbic epileptogenesis has recently drawn attention to the connectivity between the nuclei of the thalamus and limbic structures. Thalamo-limbic circuits are thought to regulate limbic seizure activity whereas thalamocortical circuits are involved in the expression and generation of spike-and-wave discharges (SWDs) in the absence epilepsy models. Genetic Absence Epilepsy Rats From Strasbourg (GAERS) and WAG/Rij (Wistar Albino Glaxo from Rijswijk) are well-defined genetic animal models of absence epilepsy. We aimed to examine the duration of behavioral changes in the kindling process and the relation of SWD activity to the kindling progress in the GAERS and WAG/Rij animals. Electrodes were stereotaxically implanted into the basolateral amygdala and the cortex of rats for stimulation and recording. The animals were stimulated at the threshold for producing afterdischarges. EEG was recorded to analyze SWDs and afterdischarge durations. The seizure severity was evaluated using Racine's 5-stage scale. None of the GAERS animals reached stage 3, 4, or 5 after application of 30 stimulations. The WAG/Rij animals showed different rate of kindling, therefore they were further categorized into the kindling-resistant, slow-kindled, and rapid-kindled groups. The kindling-resistant animals demonstrated a significantly longer duration of SWDs on the first day of the experiment before kindling stimulation and shorter duration of afterdischarge than did the kindled WAG/Rij animals. Behavioral durations at stage 2 were longer in kindled Wistar and WAG/Rij animals compared to kindling-resistant WAG/Rij and GAERS. These results suggest that mechanisms involved in the generation of SWDs act as a counterbalance to the excitability induced by kindling. PMID- 17910596 TI - Development of spontaneous seizures after experimental status epilepticus: implications for understanding epileptogenesis. AB - This report examines several concepts concerning the latent period to the first convulsive seizure, subsequent increases in seizure frequency, and possible mechanisms of epileptogenesis after kainate-induced status epilepticus. Previous data concerning the latent period and seizure progression from intermittent and continuous behavioral monitoring are compared, and hypothetical mechanisms of acquired epilepsy are discussed. Data involving electrographic recordings with tethered animals or with radiotelemetry are assessed in terms of their potential for addressing different hypotheses concerning the latent period and progressive changes in seizure frequency. Experimental analyses of the time course of occurrence of spontaneous seizures are interpreted in terms of possible cellular mechanisms underlying epileptogenesis. PMID- 17910597 TI - Comparison of cortical epileptic afterdischarges in immature genetic absence epilepsy WAG/Rij rats with those in two other strains (ACI and Wistar). AB - The aim of this study was to examine the development of cortical epileptic afterdischarges (ADs) in genetic absence epilepsy WAG/Rij rats, and to compare them with two strains with minimal incidence of spike-and-wave (SW) episodes (ACI and Wistar). Epileptic ADs were elicited by stimulation of sensorimotor cortex in 12-, 18-, and 25-day-old rats of the three strains. The threshold current intensities were established for movements accompanying stimulation, for ADs of the SW type and accompanying clonic seizures and for transition into limbic type of ADs (characterized by behavioral automatisms). Individual groups were formed by 7-12 rats. There were no differences among the three strains in the thresholds for elicitation of stimulation-bound movements. In contrast, WAG/Rij and ACI rats exhibited easier elicitation of SW ADs than Wistar rats at the age of 18 and 25 days. There was no difference among the three strains in transition into the limbic type of ADs in 18- and 25-day-old rats. Lower thresholds for SW ADs in 18- and 25-day-old WAG/Rij and ACI rats in comparison with Wistar rats are in agreement with our data from adult animals as well as with development of pharmacologically induced models of absence seizures. The failure to find a specific difference between WAG/Rij rats and the other two strains might indicate a difference in generation of SW episodes and SW cortical AD. PMID- 17910598 TI - Melatonin as a naturally occurring co-substrate of quinone reductase-2, the putative MT3 melatonin membrane receptor: hypothesis and significance. AB - The nature of the MT3 melatonin receptor/binding site has been a long pondered mystery for scientists. Even though it is a presumptive membrane receptor, neither its transduction cascade nor its biological consequences, after its stimulation, have been uncovered. Moreover, solid data support the idea that the MT3 melatonin binding site is an enzyme, quinone reductase 2 (QR2), rather than a membrane melatonin receptor. Based on the data available and our preliminary studies, we hypothesize that melatonin is a co-substrate of QR2. We surmise that melatonin binds to a co-substrate binding site (MT3 binding site) donating an electron to the enzyme co-factor, flavin adenine dinucleotide (FAD). FAD can be reduced to either FADH or FADH2 while melatonin is converted to N1-acetyl-N2 formyl-5-methoxykynuramine and/or cyclic 3-hydroxymelatonin. QR2 is considered to be a detoxifying and antioxidant enzyme and its behavior changes depending on available co-substrates. As a naturally occurring substance, melatonin's levels fluctuate with the light/dark cycle, with aging and with health/disease state. As a result, these alterations in melatonin production under physiological or pathological conditions would probably influence the activity of QR2. PMID- 17910599 TI - Melatonin effect on bovine embryo development in vitro in relation to oxygen concentration. AB - Melatonin promotes mouse embryo development in vitro. An effect of melatonin on bovine embryo development is described here. Slaughterhouse derived oocytes were subjected to standard in vitro maturation and fertilization procedures. Presumptive zygotes were cultured for 2 days in CR1aaLA medium supplemented with melatonin (10(-4) m) or without melatonin (control). Culture was performed under two different gas atmospheres containing physiological (7%) or atmospheric (20%) oxygen concentrations (2x2 factorial analysis). After day 2, embryos from each treatment group developed to at least four-cell stage, were cultured without melatonin until day 10 at optimum 7% O2 atmosphere. Blastocyst formation rates of presumptive zygotes and of four-cell embryos were calculated for each group. Significant interactions between oxygen tension and the melatonin treatment were found. Out of four-cell embryos put into in vitro culture after initial incubation in medium containing melatonin, decreased blastocyst rate was observed in melatonin group (47.7%) compared with control (67.7%; P=0.0327) when lower oxygen concentration was applied. A beneficial effect of melatonin was observed in 20% O2: out of 61 embryos, 42 (68.9%) developed to the blastocyst stage after treatment in melatonin versus 32 of 63 (50.8%; P=0.0458) blastocysts that developed in control group. In conclusion, beneficial or harmful effects of melatonin on bovine embryo development in vitro were observed, depending on the oxygen tension during the treatment. PMID- 17910600 TI - Evidence for differential photic regulation of pineal melatonin synthesis in teleosts. AB - The aim of this study was to compare the circadian control of melatonin production in teleosts. To do so, the effects of ophthalmectomy on circulating melatonin rhythms were studied along with ex vivo pineal culture in six different teleosts. Results strongly suggested that the circadian control of melatonin production could have dramatically changed with at least three different systems being present in teleosts when one considers the photic regulation of pineal melatonin production. First, salmonids presented a decentralized system in which the pineal gland responds directly to light independently of the eyes. Then, in seabass and cod both the eyes and the pineal gland are required to sustain full night-time melatonin production. Finally, a third type of circadian control of melatonin production is proposed in tilapia and catfish in which the pineal gland would not be light sensitive (or only slightly) and required the eyes to perceive light and inhibit melatonin synthesis. Further studies (anatomical, ultrastructural, retinal projections) are needed to confirm these results. Ex vivo experiments indirectly confirmed these results, as while the pineal gland responded normally to day-night rhythms in salmonids, seabass and cod, only very low levels were obtained at night in tilapia and no melatonin could be measured from isolated pineal glands in catfish. Together, these findings suggest that mechanisms involved in the perception of light and the transduction of this signal through the circadian axis has changed in teleosts possibly as a reflection of the photic environment in which they have evolved in. PMID- 17910601 TI - Nap and melatonin-induced changes in hippocampal activation and their role in verbal memory consolidation. AB - Overnight sleep contributes to memory consolidation; even a short nap improves memory performance. Such improvement has been linked to hippocampal activity during sleep. Melatonin has been shown to affect the human hippocampus and to induce 'sleep like' changes in brain activation. We therefore conducted and compared two functional magnetic resonance imaging studies: the first study assessed the effect of a 2-hr mid-day nap versus an equal amount of wakefulness on a verbal memory task (unrelated word pair association); the second assessed the effect of melatonin versus placebo (both conditions without nap) on a similar task. We report that following a nap relative to wakefulness, successful retrieval-related activation in the parahippocampus is decreased. A smaller decrease is seen in wakefulness with melatonin but not placebo. In parallel, an improvement in verbal memory recall was found after a nap compared with wakefulness but not with melatonin during wakefulness compared with placebo. Our findings demonstrate effects of melatonin that resemble those of sleep on verbal memory processing in the hippocampus thus suggesting that melatonin, like sleep, can initiate offline plastic changes underlying memory consolidation; they also suggest that concomitant rest without interferences is necessary for enhanced performance. PMID- 17910602 TI - N-nitroso-melatonin releases nitric oxide in the presence of serotonin and its derivatives. AB - A novel reaction was observed between 5-hydroxytryptophan derivatives like serotonin and N-nitroso-melatonin (NOMela). This reaction decreased the concentration of serotonin by about 50% and generated initially as detectable products nitric oxide and melatonin with stoichiometrical yields. The other expected product, a serotonin-derived radical, could not be detected by electron spin resonance (ESR) spectrometry, probably because the self-decay of phenoxyl type radicals proceed at the diffusion-controlled limit. From the facts that the decay rate of NOMela corresponded very well with the nitric oxide releasing rate and that nitrite was the only thermodynamically stable nitrogen oxide-containing product, it is concluded that the NOMela-serotonin reaction proceeded quantitatively. The observed reaction might be a possibility to counteract a pharmacologically abnormal high serotonin concentration in various diseases. PMID- 17910603 TI - Long-term effectiveness outcome of melatonin therapy in children with treatment resistant circadian rhythm sleep disorders. AB - To date, there have been no prospective long-term studies of melatonin therapy in children. We report here data from a prospective follow-up study of 44 children with neurodevelopmental disabilities and treatment-resistant circadian rhythm sleep disorders (CRSD) who had participated in a placebo controlled, double blind cross-over trial of sustained-release melatonin. The follow-up study involved a structured telephone interview of caregivers every 3 months for upto 3.8 yr. The caregivers provided ratings of satisfaction, adverse effects, benefits, persistence with treatment and additional medications. Changes in melatonin dose were recorded. Open ended questions were included to capture caregivers' impressions and comments concerning melatonin therapy. Adverse reaction to melatonin therapy and development of tolerance were not evident. Better sleep was associated with reported improvement in health, behavior and learning. At the end of the study, the parental comments regarding the effectiveness of long-term melatonin therapy were highly positive. Parents whose children had sleep maintenance difficulties expressed a wish to have a commercially available controlled-release melatonin product which would promote sleep for 8-10 hr. Hypnotics for children with CRSD should be considered a second line of treatment for those who fail to respond to sleep hygiene and/or melatonin. PMID- 17910604 TI - Human malarial parasite, Plasmodium falciparum, displays capacitative calcium entry: 2-aminoethyl diphenylborinate blocks the signal transduction pathway of melatonin action on the P. falciparum cell cycle. AB - The malarial parasite senses the environment to modulate its own cycle. Knowledge of the mechanisms for regulation signaling processes at the invasion, maturation, as well as division of Plasmodium falciparum before reinvasion would represent a major breakthrough and, therefore, might open new avenues for therapy. We have previously reported that melatonin modulates the circadian rhythm of malarial parasites through the activation of phospholipase C (PLC), production of InsP3, and induction of calcium release from intracellular stores. To further investigate the molecular mechanism of melatonin's action, we have used the InsP3 modulator 2-aminoethyl diphenylborinate (2-APB) given in a culture of P. falciparum parasites. Here we show that the melatonin acts on Plasmodium cell cycle through InsP3 signaling as 2-APB blocks melatonin's effect on calcium release. The function of the InsP3 signaling can be regarded as an important event for parasite invasion and maturation process, since addition of the PLC inhibitor, U73122 into Plasmodium-infected red blood cells impairs parasite invasion in vitro. By using 8BrcAMP, we also report here that Plasmodia displays a 'capacitative calcium entry' mechanism for amplification of calcium signals throughout the cytoplasm. PMID- 17910605 TI - Pineal melatonin and the innate immune response: the TNF-alpha increase after cesarean section suppresses nocturnal melatonin production. AB - The nocturnal surge of melatonin is the endocrine expression of the circadian system and is essential for organizing the timing of various endogenous processes. Previous works suggest that, in the beginning of a defense response, the increase in circulating tumor necrosis factor-alpha (TNF-alpha) leads to a transient block of nocturnal melatonin production and promotes a disruption of internal time organization. In the present paper, the concentration of melatonin and cytokines [TNF-alpha, interferon-gamma (IFN-gamma), interleukin (IL)-2, IL-4, IL-5, IL-10, IL-12] in the colostrum (postdelivery day 3) and in the milk (postdelivery days 10, 15, 20 and 30) obtained at midday and midnight from mothers who gave birth by vaginal or cesarean section were compared. The nocturnal melatonin surge observed 3 days after vaginal delivery was absent after cesarean section. IL-12 presented no daily variation in either case, while daily variations in IFN-gamma, IL-10, IL-4 and IL-5 were observed after vaginal delivery and cesarean section. On the other hand, the increase in TNF-alpha after cesarean section resulted in suppression of the nocturnal melatonin surge. Daily variation of IL-2 was only observed after recovery of the nocturnal melatonin surge, 30 days after cesarean section. The present paper supports the hypothesis of a cross-talk between the pineal gland and the immune system, which could represent a putative immune-pineal axis. PMID- 17910606 TI - Melatonin inhibits free radical-mediated mitochondrial-dependent hepatocyte apoptosis and liver damage induced during malarial infection. AB - We showed earlier that malarial infection significantly induces liver apoptosis mediated by oxidative stress mechanisms. Thus, a nontoxic antioxidant antiapoptotic molecule may be beneficial for hepatoprotection. Melatonin remarkably prevents hepatocyte apoptosis in mice induced during malaria as indicated by caspase 3 and TUNEL assays as well as transmission electron microscopy (TEM) of the liver tissue. The mitochondrial apoptotic pathway, which plays a critical role in liver cell death during malarial infection, was almost completely suppressed by melatonin as it corrects both the overexpression of Bax and down-regulation of bcl-2 as revealed by semiquantitative RT-PCR. Fluorometric studies using JC-1 documented that melatonin also restores mitochondrial transmembrane potential (DeltaPsim) in malaria-infected mice liver. The antiapoptotic effect of melatonin is associated with its antioxidant role because melatonin protects liver from oxidative stress induced during malaria by scavenging the hydroxyl radicals, preventing the depletion of reduced glutathione, inhibiting lipid peroxidation and protein carbonyl formation. The effective antioxidant dose of melatonin to protect liver from oxidative stress during malaria is 20 times lower than that of known antioxidants, vitamin C and vitamin E. Apoptosis of hepatocytes during malarial infection is well correlated with dysfunction of the liver while melatonin offers hepatoprotective effects as indicated by different liver function tests. Thus, melatonin may well be effective in combating oxidative stress-induced apoptosis and liver damage during malaria infection. PMID- 17910607 TI - Reactions of the NO redox forms NO+, *NO and HNO (protonated NO-) with the melatonin metabolite N1-acetyl-5-methoxykynuramine. AB - The different NO redox forms, NO+, *NO and HNO (=protonated NO-), were compared for their capabilities of interacting with the melatonin metabolite N1-acetyl-5 methoxykynuramine (AMK), using NO+SbF6-, PAPA-NONOate and Angeli's salt as donors of the respective NO species. Particular attention was paid to stability and possible interconversions of the redox forms. *NO formation was followed by measuring the decolorization of 2-(trimethylammonio-phenyl)-4,4,5,5-tetramethyl imidazoline-1-oxyl-3-oxide (TMA-PTIO), at different pH values, at which NO+ is, in aqueous solution, either highly unstable (pH 7.4) or relatively stable (pH 2.0). *NO donation by PAPA-NONOate, as indicated by TMA-PTIO decolorization, was similar at either pH and 3-acetamidomethyl-6-methoxycinnolinone (AMMC) was formed as the major product from AMK, at pH 7.4 more efficiently than at pH 2.0. At pH 2.0, TMA-PTIO decolorization by NO+SbF6- was much weaker than by PAPA-NONOate, but AMMC was produced at substantial rates, whereas neither TMA-PTIO decolorization nor AMMC formation was observed with the NO+ donor at pH 7.4. As NO+ is also stable in organic, especially aprotic solvents, NO+SbF6- was reacted with AMK in acetonitrile, ethanol, butanol, and ethyl acetate. In all these cases, AMMC was the only or major product. In ethyl acetate, N1-acetyl-5-methoxy 3-nitrokynuramine (AMNK) was also formed, presumably as a consequence of organic peroxides emerging in that solvent. Presence of tert-butylhydroperoxide in an ethanolic solution of NO+SbF6- and AMK also resulted in AMNK formation, in addition to AMMC and two red-fluoresecent, to date unknown products. However, hydrogen peroxide enhanced *NO-dependent AMMC production from AMK and also from N1-acetyl-N2-formyl-5-methoxykynuramine. HNO donation by Angeli's salt (Na2N2O3) also caused AMMC formation from AMK at pH 7.4, with a somewhat lower efficiency than PAPA-NONOate, but no AMNK nor any other product was detected. Therefore, all three NO congeners are, in principle, capable of nitrosating AMK and forming AMMC, but in biological material the reaction with NO+ is strongly limited by the extremely short life-time of this redox form. PMID- 17910608 TI - Melatonin protects against common deletion of mitochondrial DNA-augmented mitochondrial oxidative stress and apoptosis. AB - Defected mitochondrial respiratory chain (RC), in addition to causing a severe ATP deficiency, often augments reactive oxygen species (ROS) generation in mitochondria (mROS) which enhances pathological conditions and diseases. Previously, we demonstrated a potent endogenously RC defect-augmented mROS associated dose-dependently with a commonly seen large-scale deletion of 4977 base pairs of mitochondrial DNA (mtDNA), i.e. the common deletion (CD). As current treatments for CD-associated diseases are rather supplementary and ineffective, we investigated whether melatonin, a potential mitochondrial protector, provides beneficial protection for CD-augmented mitochondrial oxidative stress and apoptosis particularly upon the induction of a secondary oxidative stress. Detailed mechanistic investigations were performed by using laser scanning dual fluorescence imaging microscopy to provide precise spatial and temporal resolution of mitochondrial events at single cell level. We demonstrate, for the first time, that melatonin significantly prevents CD augmented mROS formation under basal conditions as well as at early time-points upon secondary oxidative stress induced by H2O2 exposure. Thus, melatonin prevents mROS-mediated depolarization of mitochondrial membrane potential (DeltaPsim) and subsequent opening of the mitochondrial permeability transition pore (MPTP) and cytochrome c release. Moreover, melatonin prevents depletion of cardiolipin which appears to be crucial for postponing later MPTP opening, disruption of the mitochondrial membrane and apoptosis. Finally, the protection provided by melatonin is superior to those caused by the suppression of mitochondrial Ca2+ regulators including the mitochondrial Na+-Ca2) exchanger, the MPTP, and the mitochondrial Ca2+ uniporter and by antioxidants including vitamin E and mitochondria-targeted coenzyme Q, MitoQ. As RC defect-augmented endogenous mitochondrial oxidative stress is centrally involved in a variety of pathological conditions and diseases, melatonin thus may serve as a therapeutic drug to benefit many clinical conditions that involve malfunction of the mitochondria. PMID- 17910609 TI - Possible therapeutic value of melatonin in mild cognitive impairment: a retrospective study. AB - Mild cognitive impairment (MCI) is an etiologically heterogeneous syndrome characterized by cognitive impairment preceding dementia. Approximately 12% of MCI patients convert to Alzheimer's disease (AD) or other dementia disorders every year. In the present report we retrospectively examined the initial and final neuropsychological assessment of 50 MCI outpatients, 25 of whom had received daily 3-9 mg of a fast-release melatonin preparation p.o. at bedtime for 9-18 months. Melatonin was given in addition to the standard medication prescribed by the attending psychiatrist. Patients treated with melatonin showed significantly better performance in Mini Mental State Examination and the cognitive subscale of the Alzheimer's Disease Assessment Scale. After application of a battery of neuropsychological tests including Mattis' test, Digit-symbol test, Trail A and B tasks and the Rey's verbal test, better performance was found in melatonin-treated patients, except for the Digit-symbol test score which remained unchanged. Abnormally high Beck Depression Inventory scores decreased in melatonin-treated patients, concomitantly with an improvement in wakefulness and sleep quality. The results suggest that melatonin can be a useful add-on drug for treating MCI in a clinical setting. PMID- 17910610 TI - Therapeutic potential of endocannabinoid-hydrolysing enzyme inhibitors. AB - The specific protein target of delta9-tetrahydrocannabinol (delta9-THC), the main active ingredient of Cannabis sativa L., was characterized from rat brain nearly 20 years ago, and several endogenous compounds and proteins comprising the endocannabinoid (eCB) system have since been discovered. It has become evident that the eCB system consists of at least two cannabinoid receptors (i.e. the CB1 and CB2 receptors), in addition to their endogenous ligands (the eCBs) and several enzymes involved in the biosynthesis and catabolism of the eCBs. The two well-established eCBs, N-arachidonoylethanolamide (AEA) and 2 arachidonoylglycerol (2-AG), are produced by neurons on demand, act near their sites of synthesis and are effectively metabolized by fatty acid amide hydrolase (FAAH) and monoglyceride lipase (MGL), respectively. Inhibitors specifically targeting these enzymes could offer novel therapeutic approaches (e.g. for the treatment of pain and movement disorders). This MiniReview summarizes the literature concerning the potential therapeutic potential of FAAH and MGL inhibitors. PMID- 17910611 TI - Action of celecoxib on hepatic metabolic changes induced by the Walker-256 tumour in rats. AB - The purpose of the present work was to investigate the influence of celecoxib on some hepatic metabolic parameters affected by the Walker-256 tumour in rats. Celecoxib was administered daily (5-50 mg/kg body weight) beginning at the day in which the tumour cells were inocculated. At day 14, the liver was isolated and perfused in order to measure alanine transformation, glycolysis and arginine transformation. Maximal reduction of tumour growth (75%), accompanied by an almost normal weight gain, was attained with a celecoxib dose of 12.5 mg/kg. Diminution of glucose utilization (glycolysis) and inhibition of gluconeogenesis and ureogenesis from alanine caused by the tumor were totally reversed by celecoxib. Oxygen uptake by the liver was also normalized by the drug. Hepatic arginine transformation, which is normally enhanced in rats bearing the Walker 256 tumour, remained elevated in celecoxib-treated animals. It was concluded that preservation of gluconeogenesis and normalization of hepatic glucose utilization can explain, partly at least, the clinical improvement of cancer patients treated with the drug. The lack of action of celecoxib on arginine hydrolysis might indicate that reduction in polyamine synthesis is not a factor contributing to the diminished tumour growth. PMID- 17910612 TI - Protective effects of sasanquasaponin on injury of endothelial cells induced by anoxia and reoxygenation in vitro. AB - The protective effects of sasanquasaponin, an effective compound from Chinese traditional herbs, on ischaemia and reperfusion injury in mouse hearts have been suggested through modulation of intracellular Cl(-) homeostasis. The effects of sasanquasaponin on injury of endothelial cells, however, induced by anoxia and reoxygenation remain unknown. Therefore, the present study attempted to observe the effects of sasanquasaponin on anoxia and reoxygenation injury in endothelial cells and investigate its putative mechanisms. Human umbilical vein endothelial cells (HUVECs) were exposed to normoxia or anoxia and reoxygenation in the absence or presence of sasanquasaponin (10.0, 1.0 and 0.1 micromol/l). Lactate dehydrogenase activity was determined in cultured HUVECs supernatant, and malondialdehyde content, superoxide dismutase and glutathione peroxidase activities were measured in HUVECs by a colorimetric method. Neutrophil adhesion to HUVECs was assayed colorimetrically. The levels of intercellular adhesion molecule-1 and tumour necrosis factor-alpha were detected. The activity of nuclear factor kappa B was determined by flow cytometry. The results show that sasanquasaponin decreased the lactate dehydrogenase activity and malondialdehyde contents, and inhibited the neutrophil adhesion to HUVECs; sasanquasaponin, moreover, inhibited nuclear factor kappa B transnuclear activity, lowered tumour necrosis factor-alpha and intercellular adhesion molecule-1 expression levels. On the other hand, sasanquasaponin increased the mitochondrial superoxide dismutase and glutathione peroxidase activities. It is suggested that sasanquasaponin could protect HUVECs against anoxia and reoxygenation injury, and the protective mechanisms appear to be related to anti-lipoperoxidation and anti-adhesion. PMID- 17910614 TI - Effect of antipsychotics on creatine kinase activity in rat brain. AB - Typical and atypical antipsychotic drugs have different clinical and behavioural profiles. It is well described that inhibition of creatine kinase activity has been implicated in the pathogenesis of a number of diseases, especially in the brain. In this work, we evaluate the effect of haloperidol, clozapine, olanzapine or aripiprazole chronic administration on creatine kinase activity in brain of rats. Adult male Wistar rats received daily injections of haloperidol (1.5 mg/kg), clozapine (25 mg/kg), olanzapine (2.5, 5 or 10 mg/kg) or aripiprazole (2, 10 or 20 mg/kg). Our results demonstrate that haloperidol did not affect the enzyme activity in brain of rats. Clozapine inhibited the enzyme activity only in cerebellum and prefrontal cortex of rats. Aripiprazole did not affect creatine kinase in hippocampus, cerebellum and prefrontal cortex. The administration of 2.0 mg/kg aripiprazole did not alter creatine kinase activity, but 10.0 and 20.0 mg/kg aripiprazole activated the enzyme in striatum and cerebral cortex. Finally, the higher dose of olanzapine (10.0 mg/kg) activated the enzyme in striatum of rats. In hippocampus and cerebral cortex, we could not verify any effect of olanzapine on creatine kinase activity. The inhibitory effect of clozapine and olanzapine on creatine kinase activity in cerebellum and prefrontal cortex suggest that these drugs may impair energy metabolism in these brain areas. PMID- 17910613 TI - Combination therapy of rosiglitazone, a peroxisome proliferator-activated receptor-gamma ligand, and NMDA receptor antagonist (MK-801) on experimental embolic stroke in rats. AB - Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonists have been found to have potent anti-inflammatory actions and suggested as potential therapies for brain ischaemia. Glutamate is the most common excitatory neurotransmitter in the central nervous system and is released excessively during ischaemia. Stroke therapy will require combinations of drug classes, because no single drug class has yet been proven efficacious in human beings. The present study was conducted to assess whether N-methyl-d-aspartate (NMDA) receptor antagonist (MK-801) treatment can improve recovery from ischaemic brain injury and whether rosiglitazone, a PPAR-gamma ligand, can increase its neuroprotective effect in an embolic model of stroke. Stroke was induced in rats by embolizing a preformed clot into the middle cerebral artery. Rosiglitazone (0.1 mg/kg, intraperitoneally) and MK-801 (0.1 mg/kg, intravenously) were injected immediately after embolization. Forty-eight hours later, the brains were removed, sectioned and stained with triphenyltetrazolum chloride and analysed by a commercial image processing software programme. Rosiglitazone and MK-801 alone or in combination decreased infarct volume by 49.16%, 50.26% and 81.32%, respectively (P < 0.001). Moreover, the combination therapy significantly decreased the infarct volume when compared to any drug used alone (P < 0.05). MK 801 reduced the brain oedema by 68% compared to the control group (P < 0.05), but rosiglitazone or combination did not show any significant effect. The drugs alone or in combination also demonstrated improved neurological function, but combination therapy was more effective on neurological deficits improving. Our data show that the combination of MK-801 and rosiglitazone is more neuroprotective in thromboembolic stroke than given alone; this effect perhaps represents a possible additive effect in the brain infarction. PMID- 17910615 TI - Effects of marjoram volatile oil and grape seed extract on ethanol toxicity in male rats. AB - Natural dietary antioxidants are extensively studied for their ability to protect cells from miscellaneous damages. Marjoram volatile oil (Origanum majorana L., Lamiaceae) and grape seed extract (Vitis vinifera L., Vitaceae) are potent antioxidants. Effects of administration of marjoram volatile oil or grape seed extract on oral administration of ethanol, simultaneously, daily for 10 weeks were studied through determining epididymal spermatozoal analysis, serum testosterone level, weight and histopathological examination of testis, liver and brain. Glutathione level and lipid peroxidation content as malondialdehyde in the testis, liver and brain were measured. The repeated intake of a great amount of ethanol (10 ml/kg body weight, 25% v/v) was followed by fertility disturbances with low sperm count, impaired sperm motility and decrease in serum testosterone level. Moreover, ethanol toxicity induced significant alterations in the histological structures of the testis, liver and brain. The results revealed a significant increase in lipid peroxidation and decrease in the level of glutathione in the testis, liver and brain in the ethanol-treated group. However, co-administration of the extracts of protective plants resulted in minimizing the hazard effects of ethanol toxicity on male fertility, liver and brain tissues. It may be concluded that marjoram volatile oil and grape seed extract are useful herbal remedies, especially for controlling oxidative damages. PMID- 17910616 TI - Comparison of reactivating and therapeutic efficacy of two salts of the oxime HI 6 against tabun, soman and cyclosarin in rats. AB - The reactivating and therapeutic efficacy of two salts of the oxime HI-6 (dichloride and dimethanesulphonate) against chosen nerve agents (tabun, soman and cyclosarin) was compared in rats. The potency of both salts of HI-6 to decrease the acute toxicity of tabun, soman and cyclosarin was similar in nerve agent-poisoned rats. While the potency of HI-6 dichloride and HI-6 dimethanesulphonate to counteract acute toxic effects of tabun is rather low, both salts of HI-6 were able to decrease the acute toxicity of soman two times and acute toxicity of cyclosarin more than three times. The therapeutic efficacy of both salts of the oxime HI-6 corresponds to their reactivating potency. While the reactivating efficacy of HI-6 dichloride as well as HI-6 dimethanesulphonate against tabun was negligible, their potency to reactivate soman-inhibited acetylcholinesterase and cyclosarin-inhibited acetylcholinesterase in peripheral (blood) and central (brain) compartment was relatively high. HI-6 dichloride showed a somewhat higher potency to reactivate tabun-inhibited acetylcholinesterase in brain, and soman-inhibited acetylcholinesterase in blood and brain than HI-6 dimethanesulphonate but the differences were not significant. Thus, the replacement of dichloride anion by dimethanesulphonate anion in the oxime HI-6 does not influence the therapeutic and reactivating efficacy of the oxime HI-6 against nerve agents. In addition, the higher solubility and stability of HI-6 dimethanesulphonate in comparison with HI-6 dichloride makes it possible to increase the dose and thus, the effectiveness of the oxime HI-6 in the antidotal treatment of acute nerve agent poisonings. PMID- 17910617 TI - Protection of arsenic-induced hepatic disorder by arjunolic acid. AB - Arsenic is one of the ubiquitous environmental pollutants, which affects nearly all organ systems. The present study has been carried out to investigate the hepatoprotective role of arjunolic acid, a triterpenoid saponin, against arsenic induced oxidative damages in murine livers. Administration of sodium arsenite at a dose of 10 mg/kg body weight for 2 days significantly reduced the activities of antioxidant enzymes, superoxide dismutase, catalase, glutathione S-transferase, glutathione reductase and glutathione peroxidase as well as depleted the level of reduced glutathione and total thiols. In addition, sodium arsenite also increased the activities of serum marker enzymes, alanine transaminase and alkaline phosphatase, enhanced DNA fragmentation, protein carbonyl content, lipid peroxidation end-products and the level of oxidized glutathione. Studies with arjunolic acid show that in vitro it possesses free radical-scavenging and in vivo antioxidant activities. Treatment with arjunolic acid at a dose of 20 mg/kg body weight for 4 days prior to arsenic administration prevents the alterations of the activities of all antioxidant indices and levels of the other parameters studied. Histological studies revealed less centrilobular necrosis in the liver treated with arjunolic acid prior to arsenic intoxication compared to the liver treated with the toxin alone. Effects of a known antioxidant, vitamin C, have been included in the study as a positive control. In conclusion, the results suggest that arjunolic acid possesses the ability to attenuate arsenic-induced oxidative stress in murine liver probably via its antioxidant activity. PMID- 17910618 TI - Pharmacokinetics of two randomized trials evaluating the safety and efficacy of indinavir, saquinavir and lopinavir in combination with low-dose ritonavir: the MaxCmin1 and 2 trials. AB - Our objective was to identify possible differences in protease inhibitor plasma concentrations between and within three protease inhibitor regimens (indinavir, saquinavir and lopinavir all in combination with low-dose ritonavir) and to relate these differences to safety and efficacy. Data originated from pre-defined pharmacokinetic substudies within two randomized 48-week trials evaluating the safety and efficacy of three protease inhibitor regimens. At weeks 4 and 48, plasma was collected and minimum drug plasma concentrations, C(min), were obtained. Out of 656 randomized patients, 283 patients had available C(min) at week 4. Indinavir, saquinavir and lopinavir C(min) were high when combined with low-dose ritonavir. No significant difference in the proportion of patients experiencing treatment failure could be found according to the C(min) within any treatment arm. A saquinavir C(min) > 2000 ng/ml was associated with an increased risk of gastrointestinal grade 3 or 4 adverse events and higher total cholesterol. Overall, there were no changes in C(min) from week 4 to week 48 in patients who remained on therapy. No association between treatment failure and the C(min) could be demonstrated. Associations between high C(min) and toxicity were identified in the saquinavir arm; therefore, dose reductions may be appropriate in certain patients with C(min) several times above the minimum effective concentration. PMID- 17910619 TI - Ellagic acid prevents cisplatin-induced oxidative stress in liver and heart tissue of rats. AB - Cisplatin is one of the most active cytotoxic agents in the treatment of cancer. High doses of cisplatin have also been known to produce hepatotoxicity, and several studies suggest that supplemental antioxidants can reduce cisplatin induced hepatotoxicity. The present study was designed to determine the effects on the liver and heart oxidant/antioxidant system and the possible protective effects of ellagic acid on liver and heart toxicity induced by cisplatin. The control group received 0.9% saline; animals in the ellagic acid group received only ellagic acid (10 mg/kg); animals in the cisplatin group received only cisplatin (7 mg/kg); animals in cisplatin + ellagic acid group received ellagic acid for 10 days after cisplatin. The rats were killed at the end of the treatment period. Malondialdehyde (MDA) and glutathione (GSH) levels, glutathione peroxidase (GSH-Px) and catalase (CAT) activities were determined in liver and heart tissue. While administration of cisplatin increased the MDA levels in liver and heart tissues, it decreased the GSH, GSH-Px and CAT in these samples when compared to the control group. The administration of ellagic acid to cisplatin treated rats decreased the MDA levels, and increased GSH, GSH-Px and CAT in these samples. Cisplatin caused marked damages in the histopathological status of liver and heart tissues. These damages were ameliorated by ellagic acid administration. In conclusion, ellagic acid may be used in combination with cisplatin in chemotherapy to improve cisplatin-induced oxidative stress parameters. PMID- 17910620 TI - The in vitro inhibitory potential of trade herbal products on human CYP2D6 mediated metabolism and the influence of ethanol. AB - The six commonly used trade herbal products, St. John's wort, common valerian, common sage, Ginkgo biloba, Echinacea purpurea and horse chestnut, and ethanol, were investigated for their in vitro inhibitory potential of cytochrome P450 2D6 (CYP2D6)-mediated metabolism. Herbal components were extracted from commercially available products in a way that ensured the same composition of constituents in the extract as in the original trade products. c-DNA baculovirus expressed CYP2D6 was used with dextromethorphan as substrate. Quinidine was included as a positive control inhibitor. A validated high performance liquid chromatography methodology was used to quantify the formation of dextrorphan (product of dextromethorphan O demethylation). Ethanol showed a biphasic effect on CYP2D6 metabolism, increasing initially the CYP2D6 activity with 175% of control up to a concentration of 1.1%, where after ethanol linearly inhibited the CYP2D6 activity. All the investigated herbs inhibited CYP2D6 activity to some extent, but only St. John's wort, common sage and common valerian were considered possible candidates for in vivo clinically significant effects. They showed IC50 values of 0.07 +/- 7 x 10(-3) mg/ml, 0.8 +/- 0.05 mg/ml and 1.6 +/- 0.2 mg/ml, respectively. St. John's wort inhibited CYP2D6-mediated metabolism in an uncompetitive manner, while common valerian and common sage in a non-competitive manner demonstrated interherb differences in inhibition patterns and differences when compared to the more homogenous competitive inhibitor quinidine. Common valerian was the only herb that showed a mechanistic inhibition of CYP2D6 activity and attention should be paid to a possible toxicity of this herb. PMID- 17910623 TI - Interactions between soil and tree roots accelerate long-term soil carbon decomposition. AB - Decomposition of soil organic carbon (SOC) is the main process governing the release of CO(2) into the atmosphere from terrestrial systems. Although the importance of soil-root interactions for SOC decomposition has increasingly been recognized, their long-term effect on SOC decomposition remains poorly understood. Here we provide experimental evidence for a rhizosphere priming effect, in which interactions between soil and tree roots substantially accelerate SOC decomposition. In a 395-day greenhouse study with Ponderosa pine and Fremont cottonwood trees grown in three different soils, SOC decomposition in the planted treatments was significantly greater (up to 225%) than in soil incubations alone. This rhizosphere priming effect persisted throughout the experiment, until well after initial soil disturbance, and increased with a greater amount of root-derived SOC formed during the experiment. Loss of old SOC was greater than the formation of new C, suggesting that increased C inputs from roots could result in net soil C loss. PMID- 17910624 TI - Improvement of fibrosis in a patient with chronic myeloproliferative disease. PMID- 17910625 TI - Bone marrow fibrosis: pathophysiology and clinical significance of increased bone marrow stromal fibres. AB - In bone marrow biopsies, stromal structural fibres are detected by reticulin and trichrome stains, routine stains performed on bone marrow biopsy specimens in diagnostic laboratories. Increased reticulin staining (reticulin fibrosis) is associated with many benign and malignant conditions while increased trichrome staining (collagen fibrosis) is particularly prominent in late stages of severe myeloproliferative diseases or following tumour metastasis to the bone marrow. Recent evidence has shown that the amount of bone marrow reticulin staining often exhibits no correlation to disease severity, while the presence of type 1 collagen, as detected by trichrome staining, is often associated with more severe disease and a poorer prognosis. It was originally thought that increases in bone marrow stromal fibres themselves contributed to the haematopoietic abnormalities seen in certain diseases, but recent studies suggest that these increases are a result of underlying cellular abnormalities rather than a cause. A growing body of evidence suggests that increased deposition of bone marrow stromal fibres is mediated by transforming growth factor-beta and other factors elaborated by megakaryocytes, but it is likely that other cells, cytokines and growth factors are also involved. This suggests new avenues for investigation into the pathogenesis of various disorders associated with increased bone marrow stromal fibres. PMID- 17910626 TI - Platelet glycoprotein VI-related clinical defects. AB - Human patients with defects associated with the platelet collagen receptor, glycoprotein (GP)VI, are rare and usually described as having a mild bleeding disorder. However, here we review clinical profiles of patients with familial or acquired GPVI defects, revealing the bleeding defect is often severe and associated with immune dysfunction. GPVI is a member of the immunoreceptor family, and co-expressed on platelets with Fc receptor gamma-chain (FcRgamma). Ligand binding to GPVI leads to activation of platelet integrins, in particular alpha(IIb)beta(3) that mediates platelet aggregation; and activation of endogenous platelet metalloproteinases resulting in ectodomain shedding and release of a soluble GPVI fragment. Increasing evidence supports the functional importance of GPVI/FcRgamma in thrombus formation at arterial shear rates, and expression levels of platelet GPVI may be a marker of thrombotic risk. Over the past 20 years, patients have been reported with GPVI-related defects involving: (i) an acquired deficiency, resulting from (a) anti-GPVI autoantibodies or (b) other causes; or (ii) a congenital deficiency, where (c) GPVI is not expressed or (d) is expressed in a dysfunctional form with defective signalling to alpha(IIb)beta(3). Clinical consequences of GPVI-related defects may be uniquely informative about the role of platelet GPVI in health and disease. PMID- 17910627 TI - Flexible and dynamic organization of bone marrow stromal compartment. AB - The bone marrow mesenchymal compartment contains putative stem/progenitors of skeletal tissue components such as bone, cartilage, haematopoiesis-supporting stroma and adipocytes. Previously appreciated as vital to the support of haematopoiesis, these cells have also been recently recognized as having significant immunomodulatory properties with implications for allogeneic haematopoietic cell transplantation. Despite having been studied for more than three decades and currently being used in different clinical settings, their biology remains elusive. The aim of this review is to critically analyse the field of mesenchymal stem/progenitor cell biology, in respect of their relationship with other mesenchymal cell-types. Several issues concerning lineage commitment and inter-conversion potential between different mesenchymal cell types are reviewed. PMID- 17910628 TI - The histone deacetylase inhibitor, PXD101, potentiates bortezomib-induced anti multiple myeloma effect by induction of oxidative stress and DNA damage. AB - Clinical trials have shown the high anti-myeloma activity of the proteasome inhibitor bortezomib. The present study examined the activity of bortezomib combined with PXD101, a histone deacetylase inhibitor, against multiple myeloma (MM) and osteoclastogenesis. Treatment of myeloma cell lines with combinations of bortezomib and PXD101 led to synergistic inhibition of proliferation and induction of cell death. The combination significantly decreased the viability of primary human CD138(+) myeloma cells but not of bone marrow mononuclear cells. Further studies showed a dose-dependent activation of caspases-3, -8 and -9 and nuclear fragmentation in myeloma cells. Bortezomib/PXD101 treatment markedly triggered reactive oxygen species (ROS) generation that was accompanied by p53, H2A.X and p38-mitogen-activated protein kinase phosphorylation. ROS generation could be blocked by the free radical scavenger N-acetyl-L-cysteine. The combination of bortezomib and PXD101 also resulted in synergistic inhibition of osteoclast formation. In conclusion, bortezomib and PXD101 have different molecular targets. The combination induces cell death in myeloma cells via ROS mediated DNA damage and also inhibits osteoclastogenesis. Therefore, this study provides the rationale for the clinical evaluation of bortezomib combined with PXD101 in patients with MM. PMID- 17910629 TI - Risk factors for development of a second lymphoid malignancy in patients with chronic lymphocytic leukaemia. AB - Previous studies suggested that patients with chronic lymphocytic leukaemia (CLL) are at a three- to fivefold increased risk of developing a second lymphoproliferative disorder (LPD). This observational cohort study used the Mayo Clinic CLL Database to identify factors associated with developing a second LPD. A second LPD was identified in 26 (2.7%) of 962 CLL patients during a median follow-up of 3.3 years. Diffuse large B-cell lymphoma was the most common subtype of secondary LPD (12 of 26 cases). Patients previously treated for CLL had a trend toward higher prevalence of second LPD (4%) compared with previously untreated patients (2%; P = 0.053). More strikingly, patients treated with purine nucleoside analogues (PNA) had a significantly increased risk of subsequent second LPD (5.2%) compared with patients who had not received PNA (1.9%; P = 0.008). No statistically significant association was observed between risk of second LPD and other CLL characteristics (ZAP-70, CD38, IgV(H) mutation status or cytogenetic abnormalities). In this series, prior treatments with PNA or anthracyclines were the only significant factors associated with risk of developing a second LPD in patients with CLL. Physicians should strictly adhere to established criteria to initiate treatment for CLL patients who are not participating in clinical trials. PMID- 17910630 TI - RUNX1 gene mutation in primary myelodysplastic syndrome--the mutation can be detected early at diagnosis or acquired during disease progression and is associated with poor outcome. AB - Mutations of Runt-related transcription factor 1 (RUNX1) have been detected in patients with myelodysplastic syndrome (MDS). However, the prognostic implication of RUNX1 mutations in primary MDS is limited. The stage of the disease at which the mutations are acquired and whether they persist during the disease course also remain unclear. We analysed mutations of RUNX1 exons 3-8 in 132 patients with primary MDS and correlated the results with clinical features. Serial studies were performed during the follow-up period. Sixteen patients (12%) had RUNX1 mutations at the time of diagnosis. All RUNX1 mutations that were detected at diagnosis remained unchanged during the clinical course. Two other patients acquired RUNX1 mutations at leukaemic transformation 34 months and 35 months after the diagnosis of MDS. Patients with RUNX1 mutations at diagnosis had higher neutrophil counts and higher frequency of -7/7q deletion than those without. Furthermore, RUNX1 mutation was closely associated with a short overall survival (P = 0.039). This is the first report to demonstrate that RUNX1 mutation can not only be detected early at diagnosis but also acquired during disease progression and is associated with poor prognosis in patients with primary MDS. It may play a role in the development and progression of a subset of primary MDS. PMID- 17910631 TI - Phase I/II clinical trial of sequential subcutaneous and intravenous delivery of dendritic cell vaccination for refractory multiple myeloma using patient-specific tumour idiotype protein or idiotype (VDJ)-derived class I-restricted peptides. AB - Fifteen multiple myeloma (MM) patients who had failed maintenance therapy after tandem autologous stem cell transplantation underwent anti-idiotype (Id) vaccination with dendritic cells (DCs). CD14(+)-derived DCs were loaded with the autologous Id as whole protein (=6) or Id-derived class I-restricted peptides (=9) and keyhole limpet hemocyanin (KLH). Vaccination consisted of three subcutaneous (sc) and two intravenous injections of increasing DC doses at 2 weeks interval. DC therapy was well tolerated. Most patients developed both humoral and T-cell responses to KLH, suggesting immunocompetence. Eight of 15 patients developed an Id-specific T-cell proliferative response, 8/15 increased interferon-gamma-secreting T cells and 4/15 showed an Id-positive delayed-type hypersensitivity test. Anti-Id cytotoxic T-lymphocyte precursors increased after DC vaccination in 2/2 evaluable patients. A more robust T-cell response was observed after sc DC injections and increased Id-specific T-cell proliferation was found up to 1 year after vaccination. VDJ-derived peptides were as effective as the whole protein in stimulating T-cell responses. Clinically, 7/15 patients have stable disease after a median follow-up of 26 months, one patient achieved durable partial remission after 40 months, and seven patients progressed. In conclusion, sc injections of cryopreserved Id-pulsed DCs were safe and, in contrast with intravenous administrations, induced anti-MM T-cell responses. PMID- 17910632 TI - Pralatrexate, a novel class of antifol with high affinity for the reduced folate carrier-type 1, produces marked complete and durable remissions in a diversity of chemotherapy refractory cases of T-cell lymphoma. AB - T-cell lymphomas (TCLs) are characterised by poor responses to therapy with brief durations of remissions. An early phase study of pralatrexate has demonstrated dramatic activity in patients with relapsed/refractory disease. Of the first 20 lymphoma patients treated, 16 had B-cell lymphoma and four had refractory aggressive TCL. All four patients with TCL achieved a complete remission. Patients with B-cell lymphoma achieved stable disease at best. For each TCL patient, the response was more durable than their best response with chemotherapy. This early experience is the first to document this unique activity of pralatrexate in TCL. PMID- 17910633 TI - Thalidomide maintenance following high-dose therapy in multiple myeloma: a UK myeloma forum phase 2 study. AB - Thalidomide maintenance has unresolved issues regarding dosage and toxicity. We evaluated this in five dose cohorts in 100 patients. At a median follow-up of 32.3 months, 23 patients had stopped thalidomide for disease progression, 54 for side effects. 3-year overall and progression-free survival was 76% and 41% respectively. Dosage did not influence disease outcome but greatly affected toxicity. Fifteen patients converted from partial remission to complete remission on thalidomide at a median of 13.5 months. Maintenance doses >200 mg were largely unachievable and peripheral neuropathy was the main toxicity. Lower doses enabled more patients to stay on the drug for a useful period of time. PMID- 17910634 TI - Stimulation of new bone formation by the proteasome inhibitor, bortezomib: implications for myeloma bone disease. AB - Impaired bone formation contributes to the lack of bone healing in multiple myeloma and there is a need for agents with bone anabolic properties to reverse the bone deficit in patients. Bortezomib, a proteasome inhibitor with antitumour efficacy in myeloma patients, enhanced new bone formation in mouse calvarial cultures; this effect was blocked by dickkopf 1(Dkk1), an antagonist of Wnt signalling implicated in myeloma bone disease. Bortezomib inhibited Dkk1 expression in calvariae and bone marrow-derived stromal cells, suggesting a novel mechanism by which bortezomib exerts its effects in bone. Clinical trials in patients with myeloma bone disease are needed to validate these results. PMID- 17910635 TI - Prevalence of erythrocyte haemoglobin H inclusions in unselected patients with clonal myeloid disorders. AB - Patients with clonal myeloid disorders, especially myelodysplastic syndromes (MDS), may acquire alpha-thalassaemia. To estimate the prevalence of this erythrocyte phenotype, we examined brilliant cresyl blue-stained blood smears from 201 patients with neoplastic myeloid disorders and 282 controls (195 non clonal anaemia, 62 with medical illnesses without anaemia and 25 healthy persons). Haemoglobin H inclusions were detected in 8/100 patients with MDS (8%) and 2/81 (2.5%) patients with myeloproliferative disorders, but in none of the acute leukaemia patients or controls. We conclude that the emergence of thalassaemic clones may be relatively common in the disordered marrow milieu of MDS. PMID- 17910636 TI - High nerve growth factor receptor (p75NTR) expression is a favourable prognostic factor in paediatric B cell precursor-acute lymphoblastic leukaemia. AB - Nerve growth factor (NGF) plays a pivotal role in cellular survival/death decisions with the low affinity receptor p75NTR predominately transmitting anti proliferative signals. In spite of its established role in B-cell function and identification as a prognostically favourable marker in a number of malignancies, little is known about the expression pattern and prognostic significance of p75NTR in B cell precursor-acute lymphoblastic leukaemia (BCP-ALL). p75NTR expression was prospectively studied on primary ALL-blasts in a cohort of paediatric patients with common ALL (n = 86) and preB-ALL (n = 34) treated within the Co-operative study group for childhood acute lymphoblastic leukaemia (CoALL) protocol, CoALL06-97. Flow cytometric analysis showed that almost half of the patients expressed no or negligible amounts of p75NTR (<10%). The median expression in patients expressing p75NTR beyond that threshold was 49% (range 11 100%). In patients classified as low-risk at diagnosis, p75NTR expression was significantly higher than in high-risk patients (P = 0.001). Of note, p75NTR expression was lower in the 21 patients who subsequently developed relapse compared with those remaining in remission (P = 0.038). Accordingly, relapse-free survival was significantly better in patients expressing high surface p75NTR (P = 0.041). Thus, in this prospective analysis, high p75NTR expression was a strong prognostic marker that identified a group of paediatric ALL patients with favourable outcome. PMID- 17910638 TI - Cyclosporin and plasma exchange in thrombotic thrombocytopenic purpura: long-term follow-up with serial analysis of ADAMTS13 activity. AB - We hypothesized that cyclosporin (CSA) as adjunct to plasma exchange (PE) improves the efficacy of PE in idiopathic thrombotic thrombocytopenic purpura (TTP) via suppression of the antibody inhibitor of ADAMTS13. Our preliminary findings with CSA and PE as the upfront treatment of TTP suggested that the addition of CSA to PE significantly decreased the exacerbation (disease recurrence within 30 d of the last PE) rates compared to a cohort that received corticosteroids and PE as their upfront therapy of TTP. We present an updated analysis with long-term follow-up of 18 patients with idiopathic TTP treated with concurrent CSA and PE with analysis of serial measurements of ADAMTS13 activity, antigen and inhibitor concentration in the context of clinical outcome data. Overall, 16/18 (89%) patients achieved remission, similar to historical remission rates in idiopathic TTP with PE with only one patient suffering an exacerbation. Clinical responses correlated with improvements in ADAMTS13 activity and suppression of the antibody inhibitor of ADAMTS13. These data suggest that the efficacy of CSA is at least in part related to its suppression of the antibody inhibitor of ADAMTS13 and a subsequent improvement in ADAMTS13 activity and antigen. PMID- 17910637 TI - Influence of human leucocyte antigen disparity and graft lymphocytes on allogeneic engraftment and survival after umbilical cord blood transplant in adults. AB - The dose of graft-nucleated cells and CD34(+) haematopoietic progenitor cells are predictors of allogeneic engraftment and survival in umbilical cord blood (UCB) recipients. In this single institution prospective phase II trial, flow cytometric analyses of CD34(+) progenitor and lymphocyte populations in unmodified single unit human leucocyte antigen (HLA)-disparate UCB grafts infused into 31 consecutive adults (median age 41 years, range 20-64) receiving myeloablative conditioning were compared with clinical outcomes. Median infused UCB graft-nucleated cells and CD34(+) dose was 2.2 x 10(7)/kg and 1.2 x 10(5)/kg respectively. Day to absolute neutrophil count >/=0.5 x 10(9)/l with full donor chimerism averaged 27 d (range 12-41). Univariate analyses demonstrated that UCB graft-infused cell doses of CD34(+) (P = 0.015), CD3(+) (P = 0.024) and CD34(+)HLADR(+)CD38(+) progenitors (P = 0.043) correlated with neutrophil engraftment. This same analysis did not demonstrate a correlation between CD34(+) (P = 0.11), CD3(+) (P = 0.28) or CD34(+)HLADR(+)CD38(+) (P = 0.108) cell dose and event-free survival (EFS). High-resolution matching for HLA-class II (DRB1) resulted in improved EFS (P = 0.02) and decreased risk for acute graft-versus host disease (GVHD) (P = 0.004). Early mortality (prior to post-transplant day +28) occurred in three patients, while 26 patients achieved myeloid engraftment. These results suggest that UCB graft matching at DRB1 is an important risk factor for acute GVHD and survival, while higher UCB graft cell doses of CD34(+), committed CD34(+) progenitors and CD3(+) T cells favourably influence UCB allogeneic engraftment. PMID- 17910639 TI - Fibrinogen gamma' chain carboxy terminal peptide selectively inhibits the intrinsic coagulation pathway. AB - The minor gammaA/gamma' isoform of fibrinogen contains a high affinity binding site for thrombin exosite II that is lacking in the major fibrinogen isoform, gammaA/gammaA fibrinogen. The biological consequences of gamma' chain binding to thrombin were therefore investigated. Coagulation assays, thrombin activity assays, and a primate thrombosis model were used to characterize the biological effects of the gamma' 410-427 peptide. The gamma' peptide had little effect on thrombin cleavage of the small peptidyl substrate tosyl-glycyl-prolyl-arginine-4 nitranilide acetate. However, in vitro assays demonstrated that the gamma' peptide inhibited thrombin cleavage of larger proteinaceous substrates, including fibrinogen and factor VIII. The gamma' peptide inhibited the activated partial thromboplastin time in plasma and showed greater inhibition of activated partial thromboplastin time assays than prothrombin time assays, consistent with the inhibition of factor VIII cleavage. Studies in a baboon thrombosis model showed that the gamma' 410-427 peptide inhibited fibrin-rich thrombus formation (typical of venous thrombi) and, to a lesser extent, platelet-rich thrombus formation (typical of arterial thrombi). These results indicate that binding of thrombin exosite II by the gamma' peptide has selective effects on the intrinsic pathway. PMID- 17910641 TI - Combined factors V and VIII deficiency (F5F8D) in a Chinese family due to compound heterozygosity for nonsense mutations of the LMAN1 gene. PMID- 17910640 TI - Mixed haematopoietic chimerism for sickle cell disease prevents intravascular haemolysis. PMID- 17910644 TI - Tailoring support to the individual parent of a pediatric transplant recipient. PMID- 17910642 TI - Prevalance of JAK2 V617F and exon 12 mutations in polycythaemia vera. PMID- 17910645 TI - Kidney transplantation in infants and small children. AB - Transplantation is now the preferred treatment for children with end-stage kidney disease. But not all pediatric age groups have enjoyed the same success. The number of transplants in infants and young children has lagged behind the number in older children. One reason for this is the philosophy of some centers to maintain infants on dialysis until they reach some arbitrarily determined age, at which time they would undergo a transplant. If kidney transplantation is the therapy of choice for older children with renal failure, and equivalent results can be obtained in all age groups, why should it not be offered to these youngest patients? Our center's philosophy for many years has been not to restrict transplant based on size or age. We have performed over 50 kidney transplants in infant recipients, and have shown equivalent results to those obtained in older children. Important factors in obtaining a successful outcome include the use of adult kidneys from a living donor, careful attention to operative and perioperative care, and performing the transplant early or in a preemptive fashion. The latter allows for minimizing the negative impact of uremia on physical and neurologic development in infants. PMID- 17910647 TI - Determinants of urinary calcium and uric acid excretion in children after renal transplantation. AB - HC and HU predispose healthy children to develop hematuria and nephrolithiasis. The natural history of HC and HU has not been studied in renal transplant recipients who may be at greater risk of complications. Our study investigated the prevalence of HC and HU after Tx and determined independent predictors of urinary calcium and uric acid excretion. Twenty-five pediatric transplant patients were studied between one and 12 months after Tx. Demographic data and measurements of the random Uca/cr and uric acid excretion were collected. Multivariable regression analyses were used. The median age of the patients was 10.6 yr. The prevalence of HC and HU was 20% each at one month. At 12 months, 20% had HC and 13% had HU. There were no predictors for HC; for HU, the only predictor was systolic hypertension (p = 0.03). Our data demonstrate a high prevalence of HU and HC in pediatric renal Tx recipients. The long-term clinical implication of these metabolic abnormalities remains to be elucidated in prospective trials. PMID- 17910648 TI - Assessment of risk factors for cardiovasuclar disease in pediatric renal transplant patients. AB - Pediatric renal TP recipients are at risk for CVD. We performed a cross-sectional study of the prevalence of RF for CVD in 45 long-term pediatric renal TP patients. The time since TP was 42 months. The GFR was 87.8 +/- 3.4 mL/min/1.73 m(2); 25/45 (56%) had Stage 2-4 CKD. A total of 33% had elevated SBP and 24% had high DBP; 57% had elevated SBP or DBP. A total of 20% had elevated serum CHOL levels, while 45% had high serum TG levels. A total of 42% had high HCY levels and 50% had low HCT levels. The vast majority (66.7%) had at least two RF for CVD. A total of 18.2% had abnormal post-TP echocardiography results. There was a negative correlation between GFR and SBP, DBP, serum CHOL, HCY, and BMI. There was a positive correlation between GFR and HCT. Serum CHOL was significantly lower and SBP and DBP trended lower in patients on a SF immunosuppression regimen. Similarly, SBP and DBP trended higher and CHOL was significantly higher in patients receiving SRL vs. mycophenolate mofetil. We conclude that the majority of pediatric renal TP patients exhibit multiple CVD RF. PMID- 17910650 TI - Invasive diagnostic procedures for pulmonary infiltrates in pediatric hematopoietic stem cell transplant recipients. AB - To evaluate the role of BAL, CTB, and OLB in the management of pulmonary infiltrates in pediatric HSCT recipients, we conducted a retrospective review of clinical records of pediatric HSCT recipients. Data were analyzed using Chi square for dichotomous and anova for continuous variables. Logistic regression was used to adjust confounding variables for diagnostic yield. Forty patients underwent 44 separate procedures. Infections were the prevailing cause of infiltrates with a positive diagnostic yield (96%). CTB and OLB were performed more often in patients with focal infiltrates compared with BAL (100%, 71% vs. 22%; p < 0.01). Adverse events were not significantly different across the three procedures. OLB more often yielded information that led to change in medical management (71% vs. 0%, 34%; p < 0.05) compared with CTB and BAL. Patients who had a positive diagnostic yield had no apparent survival advantage when compared with those in whom a procedure yielded no information. Logistic regression demonstrated that focal infiltrate was the only independently predictive variable for identifying a cause of pulmonary infiltrate. In conclusion, all three invasive diagnostic procedures were safe. Having a focal infiltrate was independently and significantly associated with having a positive diagnostic yield. PMID- 17910649 TI - Avoiding steroids in pediatric renal transplantation: long-term experience from a single centre. AB - We report our experience in pediatric renal transplantation avoiding steroids whenever possible. Immunosuppression consisted of an initial induction with antithymocyte globulin followed by maintenance therapy with a calcineurin inhibitor and MMF. Steroids were only given to selected patients because of the primary disease, recurrence, rejection, or PTLD. Thirty-four transplants grafted into 32 recipients between 1995 and 2005 were followed for a median of 3.5 yr (range 1-9.8). All patients survived. Graft rejection occurred in 10 cases during the first year post-transplantation and graft survival at one, five, and seven yr was 97, 88 and 88%, respectively. Steroids were given to half of the patients (n = 16); in nine cases due to rejection. Only four patients (13%) were continuously on steroids. Calculated GFR at one to five yr post-transplant were 73, 74, 68, 64, and 70 mL/min/1.73 m(2). Unfortunately PTLD occurred in three patients, but all survived with functioning grafts. Accordingly, our findings indicate that steroid avoidance in pediatric renal transplantation is possible with good results with respect to acute graft rejection as well as long-term graft survival. PMID- 17910651 TI - Early steroid withdrawal in pediatric renal transplant on newer immunosuppressive drugs. AB - Steroids have been a cornerstone in renal transplant immunosuppression. New immunosuppressive drugs have led to protocols using early steroid withdrawal or complete avoidance. A prospective protocol in 23 pediatric renal transplant (ages 2-14 yr) who received decreasing steroid doses stopping at day 7 post-Tx, FK, and MMF were compared with a CsA, AZT, historically matched steroid-based control group. Basiliximab was used in two doses. Anthropometric, biochemical variables, AR rates, and CMV infection were evaluated and compared using Student's t-test and regression analysis. A better growth pattern was seen in steroid withdrawal group. GFR rate and serum glucose were similar in both groups. Total serum cholesterol levels were significantly lower in steroid withdrawal group. The incidence of AR at 12 months was 4.3% in steroid withdrawal group vs. 8.6% in steroid-based group (p = ns). No difference in CMV infection was observed. Hemoglobin levels were low during the first months in both groups; reached normal values after six months. SBP became higher at 12 months in steroid-based group. Patient and graft survival was 98% in both groups at one-yr post-transplant. Early steroid withdrawal was efficacious, safe, and did not increase risk of rejection, preserving optimal growth, renal function, and reducing cardiovascular risk factors. PMID- 17910652 TI - Epstein-Barr virus DNA load and seroconversion in pediatric renal transplantation with tacrolimus immunosuppression. AB - EBV infection is one of major complications arising in pediatric patients who have undergone renal transplantation. A strong correlation between the grade of immunosuppression and the development of PTLD, one of the most severe EBV associated diseases, has been recognized. In this study, we monitored the serologic profile in conjunction with peripheral blood EBV-DNA load of 32 children who underwent renal transplantation with tacrolimus as an immunosuppressant. Six patients were EBV-seronegative (EBV-) before the transplantation, and the mean DNA load in the EBV- group was significantly higher than that in the EBV-seropositive (EBV+) group. Seroconversion occurred in five of these patients in a mean period of 22 weeks after the transplantation. The EBV DNA load in the EBV+ group was maintained at a low level for a year, whereas it increased rapidly to over 1 x 10(5) copies/mL in two patients in the EBV- group three to seven months after the transplantation, which corresponds to the timing of seroconversion, and one of them developed PTLD. These observations suggest that the close monitoring of the EBV-DNA load, along with longitudinal observation of seroconversion, is essential in pediatric renal transplantation, particularly for younger children who are more likely to be EVB-. PMID- 17910653 TI - Italian experience of pediatric liver transplantation. AB - The SIGENP Group has created an Italian Liver Transplantation database. The study considers all patients under 18 yr of age on the waiting list or transplanted between 1984 and 2005. Demographic and clinical data were collected and a descriptive analysis was conducted. Kaplan-Meier survival curves were calculated and Cox's proportional-hazards regression analysis were performed to identify predictors of death after transplantation. Twenty-two Italian centers took part and data were collected on 622 cases: only 53.8% of the transplants performed up until 1998 were carried out in Italy, while this was true of 97.7% of the operations performed between 1999 and 2005. Recipient survival curve analysis revealed one-, two- and five-yr survival rates of 88, 87 and 84%, respectively, and a significant improvement in survival after 1998 (p = 0.0322). Cox's analysis identified the following risk factors for death after liver transplantation, i.e. transplantation before 1998, neoplasms or fulminant hepatic failure as indications, being in intensive care at the time of transplantation and retransplantation. The center where the transplant is performed also revealed an influence on patient survival. Thanks to a better patient follow-up and more cooperation between specialists, the mean survival after liver transplantation is improving and Italian children can be transplanted in Italy. PMID- 17910654 TI - Growth curves of pediatric patients with biliary atresia following living donor liver transplantation: factors that influence post-transplantation growth. AB - We evaluated the growth curves of children with BA after LDLT, and identified factors influencing growth velocity one-yr after LDLT (DeltaZ). The clinical data of 51 children with BA, who had an LDLT at our center from 2001 to 2005, were retrospectively reviewed. The Z scores for height and weight, and DeltaZ were studied. The correlation between DeltaZ and various clinical factors was evaluated statistically. Multivariate stepwise analyses were performed for DeltaZ. The average height and weight Z scores at the time of LDLT were -1.34 +/- 1.36 (+/-s.d.) and -0.78 +/- 1.15, respectively. Among 30 BA recipients with stable liver function after transplant, weight returned to normal one-yr post transplantation. However, height did not return to normal even by the third post transplantation year. On multivariate analyses, 73% of the variance in height DeltaZ could be accounted for by factors such as standardized height at the time of LDLT (proportion of variance: 38%), number of steroid pulse treatments (17%), donor age (10%), and the presence of HVS (9%). Fifty-four percentage of the variance in weight DeltaZ could be accounted for by factors such as standardized weight at the time of LDLT (37%) and the total steroid dose given (17%). Height and weight status at the time of LDLT likely have the strongest impact on DeltaZ. Additional factors include steroid exposure, age of the living donor, and presence of HVS, all of which should be considered to improve post transplantation growth. PMID- 17910655 TI - Tc-99m DTPA renography in children following renal transplantation: its value in the evaluation of rejection. AB - A retrospective analysis of the value of Tc-99m DTPA DRS in children requiring renal biopsy following transplantation. Thirty-one children following renal transplantation with possible rejection underwent thirty-nine DRS and biopsy within a 72-h period and clinical followed up for 12 months. The biopsy was classified according to the Banff 97. The DRS assessed semi-quantitatively images of renal perfusion and filtration, and the balance between these two images. The clinical notes were reviewed. Based on the biopsy results 15 children had acute rejection, three children chronic rejection, nine children a mixed appearance of both acute and chronic rejection while 12 children had no rejection. Based on the long-term clinical outcome, the DRS had an overall sensitivity of 76% and specificity of 86%. While renal biopsy remains the gold standard for the diagnosis of rejection, if the perfusion and filtration phases of the DRS are analysed separately and the results integrated, there is a possibility of suggesting that acute rejection is not the cause of the increase in creatinine. The DRS provides useful information to the nephrologist when taken in conjunction with the biopsy result and other investigations. PMID- 17910656 TI - Sirolimus in chronic allograft nephropathy in pediatric recipients. AB - CAN is a common cause of late graft loss. Nephrotoxicity due to CNIs is known to contribute to CAN. We retrospectively evaluated the efficacy and safety of SRL in pediatric renal Tx recipients showing CAN in their allograft biopsy. Twenty-one patients aged 10.4 +/- 4.6 yr at Tx time receiving CNIs as primary immunosuppression were converted to SRL at 58.9 +/- 49.1 months after Tx, due to progressive decline of renal function and biopsy proven CAN. Mean follow-up after switch was 19.7 +/- 9.5 months. All patients received CsA as part of the immunosuppressive regimen, at a mean dose 4.4 +/- 1.2 mg/kg/day. Mean daily dose of SRL three month after conversion was 2.6 +/- 0.8 mg/body surface area/day and the mean through levels where 6.9 +/- 2.5 ng/mL. Graft biopsies showed Grade I CAN in 12 children and Grade II CAN in nine. After SRL introduction, there were neither acute rejection episodes nor graft losses. GFR improved at three months and was sustained thereafter only in children with Grade I CAN. Post-Tx time at conversion was the only significant variable between patients who had Grade I CAN and Grade II CAN (33.6 +/- 33.3 vs. 92.7 +/- 47.5 months, p = 0.003). Nine patients had no AEs, six patients had nine SAE: five diarrhea, one herpes zoster, one pancreatic pseudo cyst, one pneumonia, and one Influenza A infection; 11 patients had 13 AEs: six oral aphthous ulcers, three urinary tract infections, two herpes simplex, one lymphedema, and one nephrotic proteinuria. Significant improvement of GFR occurred in Grade I CAN group at three months from conversion and was sustained during follow-up. Those who had Grade II CAN experienced no change in GFR. The incidence of AEs and SAE is of concern and further studies are necessary to assess their relevance. PMID- 17910657 TI - Social support, coping, and psychological distress in mothers and fathers of pediatric transplant candidates: a pilot study. AB - Both parents and children report significant psychological difficulties and family disruption prior to transplantation; however, there have been fewer studies examining predictors of distress in both mothers and fathers and across multiple transplant groups. Thirty-four mothers and 22 fathers participated in this pilot study. Parents completed measures during a routine tertiary pretransplant psychological evaluation. Paired sample t-test results indicated that mothers and fathers differed significantly on specific coping strategies employed, with fathers less likely to use engagement strategies than mothers. Correlation analyses demonstrated strong associations between engagement coping strategies and less psychological distress and the reverse with disengagement coping strategies for both mothers and fathers. Social support was associated with less psychological distress for mothers, but was unrelated to distress for fathers. Using regression analyses, for mothers, lack of social support, and disengagement coping predicted poor psychological outcomes. Taken together, these results suggest that assessing specific coping strategies employed by both mothers and fathers is an essential component of the pretransplant evaluation process. This study delineates areas for intervention that impact adjustment in parents of pediatric transplant candidates. PMID- 17910658 TI - Favorable outcome with allogeneic hematopoietic stem cell transplantation in pediatric acquired aplastic anemia patients. AB - The data of allogeneic HSCT in nine children with acquired AA between June 1998 and July 2006 were analyzed retrospectively. The median duration of time to neutrophil and platelet engraftment was 18 and 25 days, respectively. None of the patients had primary graft failure. Two (22.2%) patients developed acute GVHD and of these, one (11.1%) was Grade 1, and the other (11.1%) was Grade 3. Although the study group was composed of higher risk patients, including six of nine resistant to previous immunosuppressive treatment, eight had multiple not irradiated or filtered transfusion histories and one of the cases was only 5/6 HLA-compatible with his donor, the five-yr overall and EFS was 100%, and all recipients are alive without any graft failure. This may be attributed to the dose adjusted use of ATG according to individual transfusion history and gradual tapering of CsA and cessation at least nine months after allogeneic HSCT. PMID- 17910659 TI - Development of in situ melanoma after allogeneic bone marrow transplantation in Griscelli syndrome type II. AB - GS is an uncommon autosomal recessive disorder characterized by pigmentary dilution of the skin and hair and in most patients by abnormal regulation of the immune system. Childhood melanoma is rare in the pediatric population. The best prognosis is achieved with early diagnosis and definitive surgical excision of melanoma. We report a case of a patient with GS type II and melanoma who was successfully treated by allogeneic bone marrow transplantation and surgical excision of the melanoma. PMID- 17910660 TI - Recurrence of non-alcoholic steatohepatitis after liver transplantation in a 13 yr-old boy. AB - Nonalcoholic steatohepatitis (NASH) is the most severe form of non-alcoholic fatty liver disease (NAFLD). The aim of our study was to highlight NASH as a rare but possible problem in children. We present a case of 13-yr-boy with a well established diagnosis of liver cirrhosis secondary to NASH, who underwent orthotopic liver transplantation (OLT) at the age of 13 years. Six months after transplantation recurrence of NASH in the graft was diagnosed. In the treatment metformin was used with good effect. PMID- 17910661 TI - Successful HLA-identical hematopoietic stem cell transplantation in a patient with purine nucleoside phosphorylase deficiency. AB - PNP deficiency is an autosomal recessive metabolic disorder characterized by severe combined immunodeficiency, autoimmune hemolytic anemia, and by a complex of neurologic manifestations including ataxia, developmental delay, and spasticity. PNP protein catalyzes the phosphorolysis of deoxyinosine and deoxyguanosine. It is found in most tissues of the body but is expressed at the highest levels in lymphoid tissues. This tissue distribution explains why the lymphoid system is predominantly affected in PNP deficiency. We describe a five yr-old boy with muscular hypertonia, impaired growth, autoimmune hemolytic anemia, and neutropenia who underwent HSCT from his HLA-identical sister. One yr post-HSCT, the boy developed normal immunological functions, and his neurological status improved. PMID- 17910662 TI - Pseudotumoral azygos and paraesophageal varices of posterior mediastinum in a 15 month-old infant: a case report. AB - A case of azygos and paraesophageal varices presenting as a posterior mediastinal mass in a 15-month-old infant with biliary atresia is described. The patient was evaluated for living donor liver transplantation because of repeated cholangitis after Kasai operation, and plain CT scan demonstrated a mass in posterior mediastinum. The operation of mediastinal tumor resection was planned before liver transplantation in order to exclude malignant disease, however, possibility of paraesophageal varices remained. Contrast-enhanced magnetic resonance imaging clearly demonstrated azygos and paraesophageal varices in posterior mediastinum. Living donor liver transplantation was performed successfully without ligation of paraesophagogastric varices. Contrast-enhanced CT demonstrated distinctly decreased mediastinal mass one month after transplantation. PMID- 17910663 TI - BK virus-associated hemorrhagic cystitis in a pediatric lung transplant recipient. AB - BKV was first postulated to be a potential pathogen in 1971 when it was isolated in the urine of a renal transplant recipient. The pathology of BKV is generally confined to the urinary tract. In renal transplant recipients, BKV has been associated with hemorrhagic cystitis, urethral stenosis, and interstitial nephritis. Reports of BKV infection in lung transplant recipients are limited to a few case reports in adult patients. A recent report revealed that up to 32% of adult lung transplant recipients may shed BKV in their urine without symptoms or renal dysfunction. To our knowledge, there are no published reports of pediatric lung transplant recipients with BKV-associated hematuria. We hereby report a case of BKV-induced hemorrhagic cystitis in a pediatric lung transplant recipient. PMID- 17910664 TI - De novo urothelial carcinoma in a pediatric recipient of living donor kidney graft. AB - De novo urothelial carcinoma is relatively rare among post-transplant malignancies and never reported in pediatric kidney transplant recipients. In this paper, we reported one 12-yr-old male case with painless gross hematuria as the initial manifestation of de novo urothelial carcinoma in living donor graft pelvis. We emphasize the importance that cystoscopy and retrograde pyelography of native and transplant kidneys should be performed in all kidney transplant recipients with painless gross hematuria. PMID- 17910665 TI - Human adipose tissue-derived mesenchymal stromal cells as salvage therapy for treatment of severe refractory acute graft-vs.-host disease in two children. AB - Severe GVHD is a lethal complication to ASCT, and a number of approaches are therefore being evaluated. Recently, Le Blanc et al. reported a case of grade IV therapy-resistant acute GVHD of the gut and liver that showed rapid improvement after infusion of mesenchymal stem cells. Here we describe two pediatric patients who developed severe refractory acute GVHD following ASCT and were successfully treated with AMSC from HLA-mismatched unrelated donors. PMID- 17910666 TI - Hyperacute graft-vs.-host disease after related HLA-identical umbilical cord blood transplantation. AB - haGVHD has been described following bone marrow and peripheral blood stem cell transplantation and in a single case who received unrelated HLA mismatched CB. An unusual case of haGVHD following HLA 6/6-matched sibling CBT in a child with AML is presented. The development of haGVHD in a fully matched CBT and without precipitating factors may suggest the role of still undefined and perhaps individual contributory factors. PMID- 17910667 TI - BK virus-associated urologic complications. PMID- 17910668 TI - [High follow-up, low follow-up, no follow-up?]. PMID- 17910669 TI - Varicella vaccination in children with atopic eczema. AB - Chickenpox in children may be complicated by local or systemic bacterial infections. Group A streptococci and S. aureus are the predominant pathogens. Children with atopic dermatitis are particularly prone to bacterial superinfection. After the introduction of universal varicella vaccination in the USA ten years ago, the number of serious bacterial soft tissue infections in children dropped significantly. Since 2004, the VZV immunization has also been included in the routine German series.Many children with atopic dermatitis have not been immunized because of concerns on the part of parents or physicians. Recent studies demonstrated the safety and efficacy of VZV vaccination in children with atopic dermatitis who appear to benefit particularly from this vaccination. PMID- 17910670 TI - Recommendations for the use of immunoapheresis in the treatment of autoimmune bullous diseases. AB - Despite the use of high-dose systemic corticosteroids in combination with other immunosuppressants, in some patients with autoimmune bullous diseases only insufficient improvement is achieved. In these cases and in acute severe disease, adjuvant immunoapheresis has been increasingly used. A consensus meeting was held in mid-2005 in Hamburg, aiming at developing guidelines for the use of immunoapheresis in the treatment of autoimmune bullous diseases. This paper summarizes the experts' recommendations. PMID- 17910671 TI - Tattoo removal--state of the art. AB - Tattooing has been around since the early beginnings of modern civilization. The discovery of selective photothermolysis at last has made it possible to remove tattoos without leaving a scar. Q-switched neodymium: yttrium-aluminum-garnet, alexandrite, and ruby lasers with pulse durations in the nanosecond domain fulfill this need. Argon or cw-CO(2) lasers as well as intense pulsed light sources should not be used since they often produce significant scarring. This article provides an overview of current laser systems. Developments leading to new tattoo inks, feedback systems to detect the absorbance characteristics of tattoo inks, dermal clearing agents, and perhaps even lasers with shorter pulse durations might improve the results in the future. PMID- 17910672 TI - Cost-effectiveness of reduced follow-up in malignant melanoma. AB - BACKGROUND: Considerable variability exists in the extent and frequency of follow up examinations for melanoma patients between different countries, generating significantly different total costs and uncertain clinical benefits. PATIENTS AND METHODS: We have analyzed the follow-up of melanoma patients under clinical and economic aspects based on the latest recommendations of the American Joint Committee on Cancer (AJCC) and the German Dermatologic Society (DDG) in the Dusseldorf cohort of 526 patients (stage IIII) during a 5-year follow-up period. Outcome measures were frequency of metastasis detection, most effective detection method, costs per detected metastasis and cost per quality-adjusted life year. RESULTS: Structured follow-up detected 17 recurrences in stages I-III. Physical examination and lymph node ultrasound were the only cost-effective methods at all stages, while laboratory studies were generally not cost-effective. The implementation of a reduced, yet medically adequate follow-up reducing chest X rays, abdominal ultrasound examinations and eliminating blood tests in early stages yielded savings of more than 100,000 euro (120,000 $) annually at a tertiary care university hospital. CONCLUSION: The implementation of a reduced follow-up for melanoma patients seems not only medically justified but also economically required without adversely affecting patient outcome. PMID- 17910673 TI - Safety and efficacy of pimecrolimus cream 1% in the daily practice: results of a patient self-observation study in patients with atopic dermatitis. AB - BACKGROUND: Pimecrolimus cream 1% has proven to be well-tolerated and effective in controlled clinical studies in patients with atopic dermatitis (AD). In a 15 week patient self-observation study, safety and efficacy was investigated in the daily practice. PATIENTS AND METHODS: 3502 patients with AD (mean age 26.2 +/- 18 years, 62% female) received pimecrolimus cream 1% from 810 physicians in the German Federal Republic. The severity of the disease was assessed at baseline, two times during the 15-week observation period and at the end of treatment. Patients recorded daily the degree of erythema and pruritus. At the end of treatment, safety and efficacy were assessed by the physician based on patient's daily records and by the patient. RESULTS: The percentage of patients with severe or massive AD decreased from 25% to 7%, whereas the percentage of patients without or with mild symptoms increased from 9% to 55%.The efficacy of treatment was rated by physicians as good or very good in 83.5% of cases and by 79% of patients. At baseline 35% of the patients were free of flares as compared to 75% at the end of therapy. Disease control was better in patients who followed the recommended treatment algorithm for pimecrolimus cream. Tolerability was mostly rated as good or very good. CONCLUSION: Treatment with pimecrolimus cream 1% for patients with AD is well-tolerated and effective in daily practice. PMID- 17910674 TI - Epidermolysis bullosa acquisita and neuroendocrine pancreatic cancer - Coincidence or patho-genetic relationship? AB - The etiology of epidermolysis bullosa acquisita (EBA) is unknown. EBA may be associated with other autoimmune systemic diseases; it also has been described in connection with different malignant tumors, showing complete remission after successful treatment of the tumor. In such cases, EBA may be regarded as a paraneo-plastic dermatosis. We detected a highly differentiated neuroendocrine pancreatic cancer in a 78-year-old woman with EBA. Even thought her tumor was completely removed and the patient has been disease-free for over seven years, a complete regression of her autoimmune bullous dermatosis could not be induced. By using intravenous immunoglobulins in combination with mycophenolate mofetil, further blister formation could be ameliorated. PMID- 17910675 TI - Hay-Wells syndrome in a child with mutation in the TP73L gene. AB - Hay-Wells syndrome is a rare form of ectodermal dysplasia, also known as AEC syndrome (Ankyloblepharon filiforme adnatum, Ectodermal effects, Cleft lip/palate). It is inherited in an autosomal dominant fashion with variable expression, featuring congenital abnormalities of skin, hair, teeth, nail, eccrine and mucous glands. We present a three-month-old boy, born to unaffected parents, with typical clinical findings of AEC syndrome. In this boy, a mutation Ile537Thr (c.1610C>T) in the sterile alpha motive (SAM) domain of the TP73L (p63) gene was detected. Because of the broad spectrum of related syndromes such as Rapp-Hodgkin syndrome, Bowen-Armstrong syndrome, CHAND syndrome and epidermolysis bullosa hereditaria, the diagnosis of AEC should be base don both clinical findings and genetic analysis. PMID- 17910676 TI - Andrology. AB - Andrology is part of dermatology in Germany, as it arose from dermatology as a subspecialty. Accordingly training in andrology is part of the curriculum for specialty certification in dermatology. All dermatologists are required to "have experience in the diagnosis of andrologic disorders and their subsequent treatment". The specialty of andrology deals with male infertility problems including questions regarding fertility prophylaxis, contraception, erectile dysfunction, disturbance in libido, ejaculation and copulation, and primary and secondary hypogonadism, as well as male aging and diseases of the male breast. Evaluation and treatment of the partner may also be necessary. Ejaculate analysis is the most important laboratory tool and each dermatologist must be qualified in its performance. PMID- 17910677 TI - [75-year-old male with persistent bilateral oedema of the eyelids]. PMID- 17910678 TI - [The traction suture--a universal tool]. PMID- 17910679 TI - Contact dermatitis. PMID- 17910680 TI - [The Museum of Wax Moulages in Zurich--current relevance for dermatology, history of medicine and the general public]. PMID- 17910681 TI - [Letter on Heidi Ulrich, Michael Landthaler, Thomas Vogt, "Granulomatosis Dermatitis from a Jellyfish"--JDDG 2007; 5: 493-495]. PMID- 17910683 TI - [Working group of Dermatologic Oncology ADO. Second-line therapy in stage IV melanoma]. PMID- 17910684 TI - Phylogeny and nucleomorph karyotype diversity of chlorarachniophyte algae. AB - Chlorarachniophytes are flagellated and/or reticulopod-forming marine algae with chlorophyll a- and b-containing plastids of secondary endosymbiotic origin. They are one of only two algal groups known to possess a "nucleomorph" (i.e. the remnant nucleus of the eukaryotic endosymbiont that donated the plastid). Apart from the recently sequenced nucleomorph genome of Bigelowiella natans, little is known about the size, structure, and composition of chlorarachniophyte nucleomorph genomes. Toward the goal of better understanding nucleomorph genome diversity, as well as establishing a phylogenetic framework with which to interpret variation in chlorarachniophyte morphology, ultrastructure, and life cycle, we are studying a wide range of chlorarachniophyte strains from public culture collections and natural habitats. We have obtained 22 new chlorarachniophyte nuclear and nucleomorph 18S rRNA gene (18S rDNA) sequences and nucleomorph genome size estimates for 14 different strains. Consistent with previous studies, all of the chlorarachniophytes examined appear to possess three nucleomorph chromosomes. However, our results suggest considerable variation in nucleomorph genome size and structure, with individual chromosome sizes ranging from approximately 90 to approximately 210 kbp, and total genome sizes between approximately 330 kbp in Lotharella amoebiformis and approximately 610 kbp in unidentified chlorarachniophyte strain CCMP622. The significance of these phylogenetic and nucleomorph karyotype data is discussed. PMID- 17910685 TI - A biochemical comparison of proteases from pathogenic naegleria fowleri and non pathogenic Naegleria gruberi. AB - Naegleria fowleri is the etiologic agent of primary amoebic meningoencephalitis (PAM). Proteases have been suggested to be involved in tissue invasion and destruction during infection. We analyzed and compared the complete protease profiles of total crude extract and conditioned medium of both pathogenic N. fowleri and non-pathogenic Naegleria gruberi trophozoites. Using SDS-PAGE, we found differences in the number and molecular weight of proteolytic bands between the two strains. The proteases showed optimal activity at pH 7.0 and 35 degrees C for both strains. Inhibition assays showed that the main proteolytic activity in both strains is due to cysteine proteases although serine proteases were also detected. Both N. fowleri and N. gruberi have a variety of different protease activities at different pH levels and temperatures. These proteases may allow the amoebae to acquire nutrients from different sources, including those from the host. Although, the role of the amoebic proteases in the pathogenesis of PAM is not clearly defined, it seems that proteases and other molecules of the parasite as well as those from the host, could be participating in the damage to the human central nervous system. PMID- 17910686 TI - Microheterogeneity and coevolution: an examination of rDNA sequence characteristics in Neoparamoeba pemaquidensis and its prokinetoplastid endosymbiont. AB - Neoparamoeba pemaquidensis, the etiological agent of amoebic gill disease, has shown surprising sequence variability among different copies of the 18S ribosomal RNA gene within an isolate. This intra-genomic microheterogeneity was confirmed and extended to an analysis of the internal transcribed spacer (ITS) region. High levels of intra-genomic nucleotide diversity (Pi=0.0201-0.0313) were found among sequenced ITS regions from individual host amoeba isolates. In contrast, the ITS region of its endosymbiont revealed significantly lower levels of intra-genomic nucleotide diversity (Pi=0.0028-0.0056) compared with the host N. pemaquidensis. Phylogenetic and ParaFit coevolution analyses involving N. pemaquidensis isolates and their respective endosymbionts confirmed a significant coevolutionary relationship between the two protists. The observation of non-shared microheterogeneity and coevolution emphasizes the complexity of the interactions between N. pemaquidensis and its obligate endosymbiont. PMID- 17910687 TI - Spliced leader RNA-mediated trans-splicing in a dinoflagellate, Karenia brevis. AB - Spliced leader (SL) trans-splicing is a form of mRNA processing originally described in parasitic kinetoplastids. During this reaction, a short RNA sequence is transferred from the 5'-end of an SL transcript to a splice acceptor site on pre-mRNA molecules. Here we report numerous mRNAs from a dinoflagellate, Karenia brevis, which contain an identical leader sequence at their 5'-terminal end. Furthermore, we have isolated a gene from K. brevis encoding a putative SL RNA containing the conserved splice donor site immediately following the leader sequence. A 1,742-bp DNA fragment encoding a K. brevis 5S gene repeat was found to encode the SL RNA gene, as well as a U6 small nuclear RNA (snRNA) gene, and binding sites for the core components of the splicesome (Sm proteins) involved in RNA splicing. Therefore the K. brevis SL RNA appears to be in a genomic arrangement typical of SL genes in a number of species known to mature their mRNAs by trans-splicing. Additionally, we show that the SL gene exists as a stable snRNA and has a predicted secondary structure typical of SL RNAs. The data presented here support the hypothesis that an SL RNA is present in K. brevis and that maturation of a percentage of mRNAs in K. brevis occurs via a trans-splicing process in which a common SL sequence is added to the 5'-end of mature mRNAs. The occurrence of SL trans-splicing in a dinoflagellate extends the known phylogenetic range of this process. PMID- 17910689 TI - Description of Deviata rositae n. sp., a new ciliate species (Ciliophora, Stichotrichia) from Argentina. AB - Soil samples were taken from a temporary pond located in Buenos Aires province, Argentina, during the dry phase in the summer of 2005. The ciliates were studied alive and after staining with protargol. Deviata rositae n. sp. measures 112-154 microm in length and 21-28 microm in width in vivo and has a vermiform body. The contractile vacuole is located in the mid-body on the left. The macronucleus is moniliform and there are 1-3 micronuclei. The oral apparatus is composed of 14-18 adoral membranelles and straight paroral and endoral membranes that never intersect each other. The somatic ciliature is arranged in four frontal cirri, one buccal cirrus, six long and slightly spiraled rows of cirri with the first right row extending up to the equatorial or sub-equatorial region, and two dorsal rows of dikinetids. This new species of Deviata primarily differs from its congeners by the number of macronuclear nodules and the number and disposition of the dorsal rows of dikinetids. PMID- 17910688 TI - Small-subunit rRNA Phylogenies Suggest That Epalxella antiquorum (penard, 1922) corliss, 1960 (ciliophora, odontostomatida) is a member of the plagyopylea. AB - The odontostomatid ciliates have remained a homogeneous order of ciliates since the 1930s when they were recognized as a monophyletic assemblage. Since that time they have been placed with the heterotrich ciliates, and more recently transferred as incertae sedis to the new "riboclass" class Armophorea. We were able to obtain the small subunit rRNA gene sequence of the odontostomatid Epalxella antiquorum (Penard, 1922) Corliss, 1960, collected from the meromictic alpine Lake Alat in Germany, in July 2005. An alignment with representatives of all 11 classes of ciliates unambiguously places the Epalxella sequence with other representatives of the class Plagiopylea with 100% support in both maximum likelihood and Bayesian analyses. Epalxella is the basal lineage with trimyemid and plagiopylid ciliates forming the two terminal sister clades. While this molecular support is strong and unambiguous, there are no obvious morphological features to unite these three clades. Thus, the class Plagiopylea must continue to be referred to as a "riboclass." Using the Epalxella sequence as a basal marker, we tentatively identified 20 environmental sequences to the terminal plagiopylean clades: eight to the genus Trimyema; four to the genus Plagiopyla; and eight to two new species, one of which might represent a new plagiopylean genus. PMID- 17910690 TI - Human serum lyses Trypanosoma brucei by triggering uncontrolled swelling of the parasite lysosome. AB - Trypanosoma brucei brucei infects a wide range of mammals, but is unable to infect humans because this subspecies is lysed by normal human serum (NHS). The phenotype of cellular lysis is debated. For some authors the lysosome undergoes osmotic swelling due to massive influx of chloride ions from the cytoplasmic compartment, but others describe multiple small cytoplasmic vacuoles and general swelling of the cellular body. Using population-level imaging of live immobilized trypanosomes throughout the lysis process, we report that specific swelling of the lysosome is a genuine and major characteristic of NHS-mediated lysis and that this phenotype is independent of the strain of trypanosomes and of NHS aging or damaging. Thus, irrespective of experimental conditions NHS reproducibly induced the swelling of the parasite lysosome. PMID- 17910691 TI - The ameba Balamuthia mandrillaris feeds by entering into mammalian cells in culture. AB - Microscopic observations of live cultures of the pathogenic ameba Balamuthia mandrillaris and mammalian cells showed that amebic feeding involved the invasion of the pseudopodia, and/or the whole ameba into the cells. The ameba, recognized by their size and flow of organelles in the cytosol, was seen to extend the tip of a pseudopodium into the cytoplasm of a cell where it moved about leaving visible damage when retracted. In rounded cells, whole amebas were seen to enter into and move around before exiting a cell and then remain quiescent for hours. The invaded mammalian cells retained their turgidity and excluded vital dyes until only their denuded nuclei remained. The cytoplasm of the cells was consumed first, then the nuclei, but not their mitotic chromosomes. The feeding pattern of four isolates of B. mandrillaris, two from humans and two from soil samples, was by amebic invasion into the mammalian cells. The resulting ameba population included cysts, amebas on the surface, and free-floating amebas as individuals or in dense-packed clusters. There was no morphologic indication of a cytopathic change in the mammalian cells before their invasion by the amebas. Feeding by cell invasion is a distinctive feature of B. mandrillaris. PMID- 17910692 TI - Application of the parametric bootstrap method to determine statistical errors in quantitative X-ray microanalysis of thin films. AB - We applied the parametric bootstrap to the X-ray microanalysis of Si-Ge binary alloys, in order to assess the dependence of the Ge concentrations and the local film thickness, obtained by using previously described Monte Carlo methods, on the precision of the measured intensities. We show how it is possible by this method to determine the statistical errors associated with the quantitative analysis performed in sample regions of different composition and thickness, but by conducting only one measurement. We recommend the use of the bootstrap for a broad range of applications for quantitative microanalysis to estimate the precision of the final results and to compare the performances of different methods to each other. Finally, we exploited a test based on bootstrap confidence intervals to ascertain if, for given X-ray intensities, different values of the estimated composition in two points of the sample are indicative of an actual lack of homogeneity. PMID- 17910693 TI - Dual-mode reflectance and fluorescence near-video-rate confocal microscope for architectural, morphological and molecular imaging of tissue. AB - We have developed a near-video-rate dual-mode reflectance and fluorescence confocal microscope for the purpose of imaging ex vivo human specimens and in vivo animal models. The dual-mode confocal microscope (DCM) has light sources at 488, 664 and 784 nm, a frame rate of 15 frames per second, a maximum field of view of 300 x 250 mum and a resolution limit of 0.31 mum laterally and 1.37 mum axially. The DCM can image tissue architecture and cellular morphology, as well as molecular properties of tissue, using reflective and fluorescent molecular specific optical contrast agents. Images acquired with the DCM demonstrate that the system has the sub-cellular resolution needed to visualize the morphological and molecular changes associated with cancer progression and has the capability to image animal models of disease in vivo. In the hamster cheek pouch model of oral carcinogenesis, the DCM was used to image the epithelium and stroma of the cheek pouch; blood flow was visible and areas of dysplasia could be distinguished from normal epithelium using 6% acetic acid contrast. In human oral cavity tissue slices, DCM reflectance images showed an increase in the nuclear-to-cytoplasmic ratio and density of nuclei in neoplastic tissues as compared to normal tissue. After labelling tissue slices with fluorescent contrast agents targeting the epidermal growth factor receptor, an increase in epidermal growth factor receptor expression was detected in cancerous tissue as compared to normal tissue. The combination of reflectance and fluorescence imaging in a single system allowed imaging of two different parameters involved in neoplastic progression, providing information about both the morphological and molecular expression changes that occur with cancer progression. The dual-mode imaging capabilities of the DCM allow investigation of both morphological changes as well as molecular changes that occur in disease processes. Analyzing both factors simultaneously may be advantageous when trying to detect and diagnose disease. The DCM's high resolution and near-video-rate image acquisition and the growing inventory of molecular-specific contrast agents and disease-specific molecular markers holds significant promise for in vivo studies of disease processes such as carcinogenesis. PMID- 17910694 TI - Quantitative STEM mass measurement of biological macromolecules in a 300 kV TEM. AB - For almost four decades, the scanning transmission electron microscope (STEM) has made significant contributions to structural biology by providing accurate determinations of the molecular masses of large protein assemblies that have arbitrary shapes and sizes. Nevertheless, STEM mass mapping has been implemented in very few laboratories, most of which have employed cold field-emission gun (FEG) electron sources operating at acceleration voltages of 100 kV and lower. Here we show that a 300 kV commercial transmission electron microscope (TEM) equipped with a thermally assisted Shottky FEG can also provide accurate STEM mass measurements. Using the recently published database of elastic-scattering cross sections from the National Institute of Standards and Technology, we show that the measured absolute mass values for tobacco mosaic virus and limpet hemocyanin didecamers agree with the known values to within better than 10%. Applying the established approach, whereby tobacco mosaic virus is added to a specimen as a calibration standard, we find that the measured molecular weight of the hemocyanin assemblies agrees with the known value to within 3%. This accuracy is achievable although only a very small fraction ( approximately 0.002) of the incident probe current of 300 kV electrons is scattered onto the annular dark field STEM detector. FEG TEMs operating at intermediate voltages (200-400 kV) are becoming common tools for determining the structure of frozen hydrated protein assemblies. The ability to perform mass determination with the same instrument can provide important complementary information about the numbers of subunits comprising the protein assemblies whose structure is being studied. PMID- 17910696 TI - Fabrication of high-quality SNOM probes by pre-treating the fibres before chemical etching. AB - A new method to fabricate high-quality fibre probes for scanning near-field optical microscopes by pre-treating the fibre before chemical etching was proposed and implemented. In the pre-treating process, the diameter of the fibre is reduced to a range of 7 -20 mum by etching one end of the fibre in the HF solution and a taper angle is formed simultaneously. Then the tapered part of the fibre is coated with a thick layer of plastic film and etched in the HF solution again. High-quality probes are obtained with an apex diameter as low as approximately 20 nm and a large cone angle. PMID- 17910695 TI - Bundles of hexagonally arranged tubules in timothy grass pollen: detection of a novel pollen component using anhydrous fixation and image analysis techniques in transmission electron microscopy. AB - Pollen from timothy grass (Phleum pratense L.) was subjected to various aqueous and non-aqueous fixation and preparation protocols for transmission electron microscopy. Only in the cytoplasm of anhydrously prepared pollen grains were conspicuous inclusions observed that range in size from less than 1 mum up to 8 or 10 mum. These bodies have so far not been described in the literature. Higher magnifications show that these inclusions consist of bundles of hexagonally arranged small tubules. In order to obtain details of the ultrastructure of this novel pollen component, TEM micrographs of ultrathin sections of hexagonally arranged tubules were analyzed using Fourier transform techniques of image analysis. It was found that the tubules form groups with quasi-periodic hexagonal arrangement, with an average centre-to-centre spacing between the neighbouring tubules of approximately 42 nm. Individual tubules are formed by 12 or 13 particles. The outer diameter of the tubules ranges between 22 and 24 nm. From our experiments, we conclude that the quasi-periodic hexagonally arranged tubules forming conspicuous cytoplasmic inclusions in dry timothy grass pollen grains are structurally similar to microtubules. PMID- 17910697 TI - Improved correction of axial geometrical distortion in index-mismatched fluorescent confocal microscopic images using high-aperture objective lenses. AB - Extracting quantitative data from microscopic volume images is straightforward when the refractive indices of the immersion medium and the mounting medium are equal. The readings of the position of the specimen stage can be directly used to measure depth and width. Imperfectly matched immersion and mounting media result in axial geometrical distortion. Linear correction of the axial distortion using the paraxial estimate of the axial scaling factor yields results that may differ as much as 4% from the actual values. From calculations based on a theoretical expression of the 3-D point-spread function in the focal region of a high aperture microscope focussing into a mismatched mounting medium, we derived axial scaling factors that result in quantitative results accurate to better than 1%. From a non-linear correction procedure, an improved formula for the paraxial estimate of the axial scaling factor is derived. PMID- 17910698 TI - Feasibility of detecting trabecular bone around percutaneous titanium implants in rabbits by in vivo microfocus computed tomography. AB - OBJECTIVES: The goal of this study was to examine the feasibility of in vivo imaging of trabecular bone around titanium implants by means of microfocus computed tomography (micro-CT) and the use of rabbits for this purpose. MATERIALS AND METHODS: Ten male rabbits type Hollander, received a titanium implant (1.7 mm diameter and 10 mm length) in the trabecular bone of the left tibia. Seven weeks later a micro-CT scan was taken. Four rabbits were used to monitor potential harmful effects from X-ray absorption until 4 weeks after scanning. A second group of six rabbits was used for testing the hypothesis that a good correlation exists between in vivo micro-CT images and histological images of trabecular bone around titanium implants. The six rabbits were scanned and sacrificed immediately. The tibias were extracted and submitted to standard histological procedures. This resulted in a total of 12 histological sections and their corresponding 12 micro-CT images. Bone area measurements were performed at the left and right side of the implant in three regions: 0-500, 500-1000 and 1000 1500 microm distance from the implant interface. Intra-class correlations (ICC) were calculated between both techniques. RESULTS: The four rabbits did not show any sign of radiodermatitis 4 weeks after scanning. In the micro-CT images of the group of six rabbits, trabeculae are visible, but not well defined, due to the presence of noise in the image. The ICC for the right implant side were 0.44 for zone 0-500 microm, 0.48 for zone 500-1000 microm and 0.40 for zone 1000-1500 microm. The ICC for the left implant side could not be calculated. CONCLUSION: A low agreement was found between the bone measurements from histology and in vivo micro-CT images. The use of the in vivo micro-CT for trabecular bone imaging around metallic implants should be restricted to track tendencies in follow-up studies. PMID- 17910699 TI - Characterization of the calcification of cardiac valve bioprostheses by environmental scanning electron microscopy and vibrational spectroscopy. AB - Bioprosthetic heart valve tissue and associated calcification were studied in their natural state, using environmental scanning electron microscopy (ESEM). Energy dispersive X-ray micro-analysis, X-ray diffraction, Fourier-transform infrared and Raman spectroscopy were used to characterize the various calcific deposits observed with ESEM. The major elements present in calcified valves were also analyzed by inductively coupled plasma-optical emission spectroscopy. To better understand the precursor formation of the calcific deposits, results from the elemental analyses were statistically correlated. ESEM revealed the presence of four broad types of calcium phosphate crystal morphology. In addition, two main patterns of organization of calcific deposits were observed associated with the collagen fibres. Energy dispersive X-ray micro-analysis identified the crystals observed by ESEM as salts containing mainly calcium and phosphate with ratios from 1.340 (possibly octacalcium phosphate, which has a Ca/P ratio of 1.336) to 2.045 (possibly hydroxyapatite with incorporation of carbonate and metal ion contaminants, such as silicon and magnesium, in the crystal lattice). Raman and fourier-transform infrared spectroscopy also identified the presence of carbonate and the analyses showed spectral features very similar to a crystalline hydroxyapatite spectrum, also refuting the presence of precursor phases such as beta-tricalcium phosphate, octacalcium phosphate and dicalcium phosphate dihydrate. The results of this study raised the possibility of the presence of precursor phases associated with the early stages of calcification. PMID- 17910701 TI - Orientational analysis of planar fibre systems observed as a Poisson shot-noise process. AB - We consider two-dimensional fibrous materials observed as a digital greyscale image. The problem addressed is to estimate the orientation distribution of unobservable thin fibres from a greyscale image modelled by a planar Poisson shot noise process. The classical stereological approach is not straightforward, because the point intensities of thin fibres along sampling lines may not be observable. For such cases, Karkkainen et al. (2001) suggested the use of scaled variograms determined from grey values along sampling lines in several directions. Their method is based on the assumption that the proportion between the scaled variograms and point intensities in all directions of sampling lines is constant. This assumption is proved to be valid asymptotically for Boolean models and dead leaves models, under some regularity conditions. In this work, we derive the scaled variogram and its approximations for a planar Poisson shot noise process using the modified Bessel function. In the case of reasonable high resolution of the observed image, the scaled variogram has an approximate functional relation to the point intensity, and in the case of high resolution the relation is proportional. As the obtained relations are approximative, they are tested on simulations. The existing orientation analysis method based on the proportional relation is further experimented on images with different resolutions. The new result, the asymptotic proportionality between the scaled variograms and the point intensities for a Poisson shot-noise process, completes the earlier results for the Boolean models and for the dead leaves models. PMID- 17910700 TI - Quantitative investigation of murine cytomegalovirus nucleocapsid interaction. AB - In this study, we quantitatively investigate the role of the M97 protein for viral morphogenesis in murine cytomegalovirus (MCMV)-infected fibroblast cells. For this purpose, a statistical analysis is performed for the spatial distribution of nuclear B-capsids (devoid of DNA, containing the scaffold) and C capsids (filled with DNA). Cell nuclei infected with either wild-type or an M97 deletion mutant were compared. Univariate and multivariate point process characteristics (like Ripley's K-function, the L-function and the nearest neighbour distance distribution function) are investigated in order to describe and quantify the effects that the deletion of M97 causes to the process of DNA packaging into nucleocapsids. The estimation of the function L(r) -r reveals that with respect to the wild type there is an increased frequency of point pairs at a very short distance (less than approximately 100 nm) for both the B-capsids as well as for the C-capsids. For the M97 deletion mutant type this is no longer true. Here only the C-capsids show such a clustering behaviour, whereas for B capsids it is almost nonexistant. Estimations of functionals such as the nearest neighbour distance distribution function confirmed these results. Thereby, a quantification is provided for the effect that the deletion of M97 leads to a loss of typical nucleocapsid clustering in MCMV-infected nuclei. PMID- 17910702 TI - Specimen-induced distortions in light microscopy. AB - Specimen-induced aberrations affect the imaging properties in optical 3D microscopy, especially when high numerical aperture lenses are used. Studies on aberrations are often properly concerned with the degradation of image quality such as compromised resolution or reduced signal intensity. Apart from these, aberration effects can also introduce geometric image distortions. The effects, discussed here are particularly strong when thick biological specimens are investigated. Using a high numerical aperture interferometer, we measured wavefront aberrations in transmission mode and quantified geometric distortions associated with specimen-induced aberrations. This assessment for a range of biological specimens allows estimation of the accuracy of spatial measurements. The results show that high-resolution spatial measurements can be significantly compromised by specimen-induced aberrations. PMID- 17910703 TI - A simple and efficient alternative to implementing systematic random sampling in stereological designs without a motorized microscope stage. AB - When properly applied, stereology is a very robust and efficient method to quantify a variety of parameters from biological material. A common sampling strategy in stereology is systematic random sampling, which involves choosing a random sampling [corrected] start point outside the structure of interest, and sampling relevant objects at [corrected] sites that are placed at pre-determined, equidistant intervals. This has proven to be a very efficient sampling strategy, and is used widely in stereological designs. At the microscopic level, this is most often achieved through the use of a motorized stage that facilitates the systematic random stepping across the structure of interest. Here, we report a simple, precise and cost-effective software-based alternative to accomplishing systematic random sampling under the microscope. We believe that this approach will facilitate the use of stereological designs that employ systematic random sampling in laboratories that lack the resources to acquire costly, fully automated systems. PMID- 17910704 TI - Recent advances in medical dermatology. AB - Collectively, new developments in the field of medical dermatology will ultimately lead to improved patient care. We review several new findings in the dermatologic literature including the following: new questions regarding the malignant potential of anti-tumor necrosis factor agents, which are widely used for the treatment of moderate to severe psoriasis as well as psoriatic arthritis; anti-interleulin-12, a promising anticytokine for the treatment of psoriasis; diagnostic advances in the detection of latent Mycobacterium tuberculosis; advances in the primary prevention of human papillomavirus and herpes zoster; and new therapeutic options with existing medications for neuropathic pain and pruritus. PMID- 17910705 TI - Lipoid proteinosis: a case series from Istanbul. AB - BACKGROUND: Lipoid proteinosis (LP) is a very rare genodermatosis. The literature on LP consists of case reports only. As we have observed 14 LP patients belonging to nine different families in the last 15 years in our practice, we decided to review all reported Turkish LP patients in this 15-year period, and noted 37 diagnosed cases. The reasons for this relatively large number of cases, the clinical features of the patients, and the associations of LP with other clinical conditions are described in this article. METHODS: Fourteen LP patients followed in our university clinic in Istanbul were scrutinized with regard to their demographic and clinical features. Diagnoses were established using clinical features, with histopathologic confirmation in 13 cases. RESULTS: All but one of the patients had a history of consanguinity, or at least a marriage of parents from the same village. Typical cutaneous signs of LP and hoarseness of the voice were observed in all patients. Two patients of the same pedigree had insulin dependent diabetes mellitus (IDDM), two patients from two different pedigrees had short stature, one patient had multinodular toxic goiter, and one patient had celiac disease. CONCLUSION: LP is not rare in Turkey as consanguineous marriage is still a social problem, especially in some rural areas. The disease is not limited to a particular geographic region in Turkey. Short stature was observed in two cases from two different families, an association not reported previously; the association of LP with IDDM in one pedigree was thought to be coincidental. PMID- 17910706 TI - Clinical and molecular analysis of NF-kappaB essential modulator in Chinese incontinentia pigmenti patients. AB - OBJECTIVE: To analyze clinical manifestation and gene of NF-kappaB essential modulator (NEMO) in 12 pediatric incontinentia pigmenti (IP) patients. METHODS: Twelve pediatric probands with three of their mothers were enrolled in this study. Physical examinations were undertaken for all patients and questionnaires requesting additional medical and developmental data were sent to the patients' families. The deletion of exon 4-10 and all 10 exons of NEMO gene were analyzed in these cases. Skin biopsy was performed in one case. RESULTS: All 15 patients had skin pigmentation abnormality and were diagnosed according to classic skin lesions. The prevalence of the dental, neurologic system, hair abnormality, and definite family history were 80.0%, 41.67%, 58.33%, and 25.0%, respectively. Histopathological examination was consistent with the diagnosis of IP with ectodermal dysplasia. In NEMO gene, deletion of exons 4-10 were noted in three cases and two of their mothers. A deletion of 19545 T in exon 6 was noted in one case and her mother. A 21690 T to C mutation in intron 8 of NEMO were found in another one case and her mother. CONCLUSION: The results suggest that skin lesion are the most prominent findings in clinics and the traditional diagnosis of IP is based on classic melanin pigmentation. Nucleotide deletion of exons 4-10 and single nucleotide mutation/polymorphism were found in these patients, which might account for etiopathogenesis of IP. PMID- 17910707 TI - Clinical characteristics of generalized idiopathic pruritus in patients from a tertiary referral center in Singapore. AB - BACKGROUND: Generalized idiopathic pruritus is a rarely studied heterogeneous condition, where patients with non-inflamed, non-pathologic skin complain of widespread pruritus. METHODS: We had administered a detailed pruritus questionnaire, based on the short form of the McGill pain questionnaire, on 75 patients with this condition. RESULTS: All 75 patients recruited with generalized idiopathic pruritus completed the questionnaire. In 92% of the patients, pruritus appeared on a daily basis. Most patients experienced pruritus at night (56%) and in the evening (61%) and 69% had difficulty falling asleep. Pruritus mainly involved the legs (79%), arms (76%) and back (68%). Accompanying symptoms were heat sensation (17%) and pain (13%). 80% had been prescribed antihistamines, of whom 56% obtained short-term relief. The sensation of itch has been reported to be crawling (33%), tickling (20%), stinging (19%) and burning (19%). Patients also reported that the itch was unbearable (73%), bothersome (72%), annoying (67%) and worrisome (45%). The worst-state VAS scores were significantly higher in patients who reported agitation (P = 0.006), difficulty in concentration (P= 0.010) and anxiety (P= 0.033). CONCLUSION: This study describes the sensory and affective dimensions of generalized idiopathic pruritus, and this questionnaire has been found to be a useful tool for evaluating pruritus in this condition. PMID- 17910708 TI - Is nitric oxide involved in the pathophysiology of essential hyperhidrosis? AB - BACKGROUND: Essential hyperhidrosis (EH) is a disorder of excessive, bilateral, and relatively symmetric sweating occurring in the axillae, palms, soles, or craniofacial region without obvious etiology. The expression of endothelial nitric oxide synthase (eNOS) in eccrine clear cells, reported by an immunohistochemical technique, has suggested that nitric oxide (NO) may play a role in the physiology of production and/or excretion of sweat in the human skin eccrine gland. AIM: To determine plasma NO levels in patients with EH and healthy controls. METHODS: We assessed the levels of plasma NO in patients with EH (n = 31) in comparison with those in age- and sex-matched healthy controls (n = 28). Total nitrite (nitrite + nitrate) was measured by a spectrophotometer at 545 nm after the conversion of nitrate to nitrite by copperized cadmium granules. RESULTS: Plasma NO levels were found to be significantly increased in EH patients in comparison with the control group (P = 0.0001). CONCLUSIONS: These findings indicate a possible role of increased plasma NO levels in the pathophysiology of EH. PMID- 17910709 TI - Aplasia cutis congenita: a rare cutaneous sign of split cord malformations. AB - BACKGROUND: Aplasia cutis congenita (ACC) is characterized by the absence of a portion of skin in a localized or widespread area at birth. It manifests usually as a solitary defect on the scalp, but sometimes may occur on the face, trunk, or limbs. ACC is most often a benign isolated defect, but can be associated with other physical anomalies or malformation syndromes. A few cases have been reported in which patients with split cord malformation (SCM) have presented with ACC. METHODS: Two patients with SCM are reported. RESULTS: Both patients presented with ACC and abnormal hair growth on their backs. Type II SCM was detected in the first patient and Type I SCM in the second. No surgical treatment was performed because the patients were neurologically intact. CONCLUSIONS: ACC may seldom manifest as a skin marker of SCM. Our patients are unique examples of SCM presenting with ACC, but not requiring surgery. It is important to recognize ACC as a cutaneous sign of SCM, and to refer these patients to radiologic evaluation as soon as possible. PMID- 17910710 TI - Staging of necrotizing fasciitis based on the evolving cutaneous features. AB - BACKGROUND: Necrotizing fasciitis is a severe soft-tissue infection characterized by a fulminant course and high mortality. Early recognition is difficult as the disease is often clinically indistinguishable from cellulitis and other soft tissue infections early in its evolution. Our aim was to study the manifestations of the cutaneous signs of necrotizing fasciitis as the disease evolves. METHODS: This was a retrospective study on patients with necrotizing fasciitis at a single institution. Their charts were reviewed to document the daily cutaneous changes from the time of presentation (day 0) through to day 4 from presentation. RESULTS: Twenty-two patients were identified. At initial assessment (day 0), almost all patients presented with erythema, tenderness, warm skin, and swelling. Blistering occurred in 41% of patients at presentation whereas late signs such as skin crepitus, necrosis, and anesthesia were infrequently seen (0-5%). As time elapsed, more patients had blistering (77% had blisters at day 4) and eventually the late signs of necrotizing fasciitis characterized by skin crepitus, necrosis, and anesthesia (9-36%) were seen. A clinical staging system was developed based on our observations. Stage migration from early to late stage necrotizing fasciitis was evident with majority of patients in stage 1 at day 0 (59%), whereas by day 4, majority had developed into stage 3 (68%). CONCLUSION: This study has demonstrated the continuum of cutaneous manifestations as necrotizing fasciitis evolves. This will help in the early recognition and intervention of this devastating condition. PMID- 17910711 TI - Suffering in psoriasis patients: its relation with illness severity and subjective well-being. AB - OBJECTIVE: The aim of the present study was to explore the relationship among subjective illness impact, subjective well-being, and psoriasis severity as assessed by dermatologists. Furthermore, subjective well-being of psoriasis patients was compared to available norm data. METHODS: Fifty-nine psoriasis patients participated in this study. The following measures were administered: (a) the Pictorial Representation of Illness and Self Measure - Revised (PRISM-R), yielding Self-Illness Separation (SIS), and Illness Perception Measure (IPM); (b) subjective health status; (c) life satisfaction, and (d) psychological well being. In addition, the Psoriasis Area and Severity Index (PASI) was determined by dermatologists. RESULTS: Psoriasis patients scored significantly below the norm data on subjective health status and psychological well-being. No differences were found concerning life satisfaction. PASI failed to correlate significantly with any of the disease impact and subjective health measures. Neither did SIS correlate significantly with any of the subjective health measures, whereas IPM was negatively associated with subjective health status, life satisfaction, and psychologic well-being. In a regression analysis with PASI as the dependent measures, none of the subjective health measures showed up as a relevant predictor. CONCLUSION: In comparison with the norm data, psoriasis patients report less subjective health status and well-being whereas their life satisfaction is not affected. There is no clear association among illness impact, subjective well-being, and illness severity as assessed by dermatologists. PMID- 17910712 TI - Screening for common dermatologic disorders amongst Israeli adolescents. AB - BACKGROUND: Dermatologic disorders amongst adolescents tend to overlap with general adult dermatology, but specific data are scarce. This study was undertaken to assess the prevalence of common skin disorders in 17-year-old Israeli military conscripts. METHODS: Military recruits who underwent medical examination over approximately 1 year were included. Dermatology specialists evaluated and classified those with suspected skin disorders into categories of suitability for military tasks. Data were computerized for analysis of prevalences. Risk ratios for each category were determined for men and women. RESULTS: Of the 94,806 adolescents, 36,511 (38.5%) women and 58,295 (61.5%) men, the most prevalent diagnoses were hyperhidrosis, multiple nevi, atopic dermatitis, keratinization disorders (mostly psoriasis), and contact dermatitis. The most prominent gender differences were in hyperhidrosis, contact dermatitis, and collagen diseases. CONCLUSION: Major dermatologic problems in adolescents are documented, and the findings may be useful for the military, employers, and general health services. PMID- 17910713 TI - DPT vaccine-induced lipoatrophy: an observational study. AB - INTRODUCTION: Diphtheria Pertussis Tetanus (DPT) vaccine is universally used in infants and children. It is generally safe and well tolerated. Local reactions such as erythema, induration, palpable nodules, and injection site abscess are well known. Injection site lipoatrophy has not been reported earlier. PATIENTS AND METHODS: Retrospective review of all cases presenting with lipoatrophy developing at injection site following DPT administration between 2000-2005 in 3 hospitals in New Delhi, India was performed. In each case, the patients were extensively evaluated for other possible causes of lipoatrophy. RESULTS: 8 infants (2 boys & 6 girls), age range 4-12 months, had presented with injection site lipoatrophy following DPT vaccination. The duration between the last injection and lipoatrophy ranged from 4 to 8 weeks. All had been administered the vaccine in the buttock instead of the thigh, as generally recommended in infants. Majority (6/8) developed lipoatrophy after the second dose. No systemic causes were found. CONCLUSION: DPT vaccine may, in rare instances, lead to injection site lipoatrophy. Inadvertent administration into the subcutaneous fat of the buttock may have been causative. Other possible mechanisms are discussed. Paramedics and general practitioners need to be educated to administer intramuscular vaccines in the thigh in infants and young children. PMID- 17910714 TI - Skin findings in internal malignant diseases. AB - PURPOSE: A skin finding may be an indicator of internal malignant diseases. In this report, we investigated the skin findings of the cases who have had internal malignancy within the last 1 month. PATIENTS AND METHODS: Seven hundred cases who were diagnosed as internal malignancy and who did not have any treatment for the malignancy were enrolled in our study between February 2002 and September 2003. A form was completed for all of the cases, including name, surname, and the carcinoma type. All cases were examined in detail, and the observed skin findings or dermatosis was recorded. RESULTS: The most frequent skin findings among the cases were tinea pedis/onychomycosis, followed by xerosis and pruritus. The skin findings in terms of frequency were determined mostly in hematological malignancies (68.96%). CONCLUSION: We would like to emphasize that the skin is an indicator of the functions of internal organs and their disorders. PMID- 17910715 TI - Imported tungiasis: a report of 19 cases and review of the literature. AB - Tungiasis is an infestation caused by penetration in the skin of the gravid female of the flea Tunga penetrans. In the period 1991-2006, 19 patients with imported tungiasis were observed at our Institute. All patients were subjected to general and dermatological examination, laboratory tests (including bacteriological examinations) and surgical excision of the lesions with histopathological examination. In all patients tetanus prophylaxis was made. All patients were followed up for at least six weeks. Thirteen patients were males (68.4%) and 6 females (31.6%). The age ranged from 3 to 71 years (average age: 37.8 years). Eleven patients (57.9%) contracted the infestation in Central and South America and 8 (42.1%) in Africa. In 17 patients (89.5%) tungiasis was localized to the feet, in one to the hands and in one to a thigh. In 13 patients (68.4%) the infestation was characterized by a single lesion; in the other 6 patients (31.6%) the lesions were two. In 15 patients (78.9%) tungiasis was characterized by a papular or nodular lesion. Crusted (2 patients), pustular (2 patients) and bullous (1 patient) lesions were also observed; furthermore, one patient presented with a plantar wart-like lesion. In only one patient bacteriological examinations showed the growth of Staphylococcus aureus and Escherichia coli. All patients healed without complications. PMID- 17910716 TI - Dermoscopic features of accessory nipples. AB - CASE 1: A 27-year-old woman presented with a 6 x 6 mm, soft, pink-brown papule located on the right axilla (Fig. 1). Dermoscopic examination showed a central scar-like white-colored area, a cleft-like appearance in the central area, and a fine pigment network in the periphery of the lesion (Fig. 2). CASE 2: A 5-year old girl presented with a 4 x 4 mm pink nodule on her left abdominal region. Dermoscopic examination showed a central scar-like white-colored area, and a fine pigment network in the periphery of the lesion. CASE 3: An 18-year-old man presented to our outpatient clinic suffering from a firm, tender, painful, 7 x 7 mm brownish pink nodule located on his left hypochondrium. Upon dermoscopic examination, a central white scar-like area was detected. A cleft-like appearance was also detected in the central region. In the periphery of the lesion, there was a fine pigment network. CASE 4: An 8-year-old girl reported to our outpatient clinic with a congenital 5 x 5 mm brownish papule on her right inguinal region. Dermoscopic examination showed a central white scar-like area, a cleft-like appearance in the central region, and a fine pigment network in the periphery of the lesion (Fig. 3). CASE 5: A 2-month-old girl was referred to our pigmented lesions clinic with a 3 x 3 mm brownish pink papule located on the right subcostal region lesion that had been noted by a pediatrician. Dermoscopic examination showed a central white area, central streak, and a very faint pigmented network at the periphery. PMID- 17910717 TI - Clinical aspects of epidermodysplasia verruciformis and review of the literature. AB - BACKGROUND: Epidermodysplasia verruciformis (EV), is an unusual genodermatosis characterized by persistent human papilloma virus infection with an autosomal recessive inheritance pattern. Clinically, it is characterized by flat wart-like lesions, scaly hypo- and hyperpigmented macules and/or patches, which resemble pityriasis versicolor, and development of early beginning nonmelanoma cutaneous carcinomas. METHODS: The epidemiological and clinical features of seven cases with EV that have been followed up for 5 years were included in the study. RESULTS: Seven cases consisted of four males and three females. All seven cases were working outdoors. Three cases were the product of consanguineous marriages. The onset of the lesions was between the ages of 1-20 years (the average age was 9.29 years). The initial appearance of cutaneous tumors were between the ages 15 34 years (average age: 21.28 years). Six of seven cases had malignant cutaneous tumors, of which histopathological examination revealed squamous cell carcinoma (SCC). Three of our cases had radiotherapy previously for the existing SCCs. These cases had more early malignant transformations. CONCLUSION: Radiotherapy applied against the SCC previously, together with sunlight exposure may cause early malignant transformation of skin lesions and the destructive tumors. PMID- 17910718 TI - Spontaneous regression of generalized eruptive histiocytosis: possible involvement of apoptosis? AB - Generalized eruptive histiocytosis (GEH) is a rare generalized non-X histiocytosis, first described in 1963 by Winklemann and Muller. Since then more than 20 patients with GEH, mainly adults but also a few children, have been reported. We report a case of GEH in a middle-aged woman with spontaneous regression, in which the ultrastructural findings of apoptosis were observed. PMID- 17910719 TI - Urethral meatal stenosis in junctional epidermolysis bullosa: a rare complication effectively treated with a novel and simple modality. AB - A 10-year-old boy was diagnosed with non-Herlitz junctional epidermolysis bullosa (nh-JEB) shortly after birth when he developed blisters on his feet and under the toenails. His younger brother was also affected by nh-JEB. Their parents were first cousins. Immunofluorescence mapping showed a blister in the lamina lucida and reduced staining with GB3 antibody against laminin V. Molecular analysis showed a homozygous mutation in the LAMC2 gene. The patient complained of intermittent inability to pass urine, causing significant suprapubic pain, anxiety, and general discomfort. He reported the development of blisters and erosions at the urethral meatus, which caused fusion of the meatal opening. In order to micturate, he had to tear apart the fused tissue, resulting in considerable pain. Physical examination revealed multiple healing erosions and atrophic scars, mainly on the legs (Fig. 1), and anonychia of most of the toenails. The walls of the urethral meatus were adherent (Fig. 2). The urethral meatus could not be opened using a moderate amount of pressure by the examining physicians. Prevention of re-stenosis of the urethral meatus was accomplished with the application of Mepilex (Molnlycke, Sweden) to the urethral meatus after each micturition, where it remained until the next episode of micturition. Since the institution of this care regimen 10 months ago, there has been no recurrence of the stenosis. PMID- 17910720 TI - alpha1-Antitrypsin deficiency presenting with panniculitis and incidental discovery of chronic obstructive pulmonary disease. AB - A 60-year-old man presented to the Emergency Department (ED) with large, painful, indurated plaques on the right thigh, left abdomen, left chest, and right chest, which began without any preceding trauma on the right thigh 3 weeks prior to presentation in the ED. He was initially treated with cefazolin 1 g three times daily as home infusions. When the lesions continued to progress, he was admitted to the hospital and placed on amoxicillin/clavulanate and vancomycin. He had a single episode of fever of 102 degrees F, but his white blood cell count and differential remained normal. An initial biopsy showed a dermal inflammatory infiltrate composed primarily of neutrophils and eosinophils with rare flame figures in the dermis. There was minimal fat seen in this biopsy. A differential diagnosis of Wells or Sweet's syndrome was entertained, and he was placed on 60 mg/day prednisone with no resolution of his symptoms. The patient's past medical history included hypertension, hyperlipidemia, peripheral neuropathy, and hiatal hernia. His family history was significant for emphysema in both parents and coronary artery disease in his father. Both of his parents smoked cigarettes. His grandfather, who was a coal miner, also had emphysema. Whilst on antibiotics and prednisone, the plaques on the patient's right thigh, right abdomen, and left chest expanded and ulcerated, draining an oily liquid (Figs 1 and 2). An incisional biopsy was obtained from his thigh. Histopathology showed a septal and lobular panniculitis with fat necrosis, neutrophils, and histiocytes (Fig. 3). Special stains for organisms were negative. Tissue sent for bacterial and fungal culture had no growth. Amylase and lipase levels were normal. Rheumatoid factor, antinuclear antibody (ANA), antineutrophil cytoplasmic antibody (ANCA), cryoglobulins, and antiphospholipid antibodies were all normal. The alpha1 antitrypsin level was low at 25 mg/dL (ref. 75-135). The alpha1-antitrypsin phenotype was PiZZ. The patient had a normal glucose-6-phosphate dehydrogenase level and was placed on dapsone 200 mg/day. The inflammation resolved and, over the course of several months, the involved areas healed with scarring. The patient denied any pulmonary complaints but, during his hospitalization, was found incidentally to have an oxygen saturation of 88% on room air. He was sent for evaluation by a pulmonologist, and pulmonary function tests revealed a mixed restrictive and obstructive pattern with a forced expiratory volume in 1 to forced vital capacity (FEV(1)/FVC) ratio of 63% of predicted. He had never smoked. He was placed on supplemental oxygen but, as his pulmonary disease has been stable, he has not been treated with intravenous antitrypsin inhibitor. PMID- 17910721 TI - Hydroa vacciniforme-like Epstein-Barr virus-associated monoclonal T lymphoproliferative disorder in a child. AB - Hydroa vacciniforme (HV) is a chronic photosensitivity disorder induced by ultraviolet radiation. Hydroa vacciniforme-like lymphoma is a rare cutaneous T cell lymphoma occurring mainly in childhood. Recent studies have demonstrated an association between chronic latent Epstein-Barr virus (EBV) infection and both the benign skin disorder and the lymphoma. The authors report a 6-year-old boy with chronic EBV infection, HV-like skin eruptions, and chronic hepatitis. Histopathologic examination of a skin biopsy specimen demonstrated epidermal ballooning degeneration and dense superficial and deep perivascular and periappendageal lymphoid cell infiltrates extending to the fat lobules. Some blood vessels in the deep plexus were infiltrated by predominantly CD4+ and TIA 1+ cytotoxic T cells. The EBV genomes were found within tissue from three skin biopsies and peripheral blood cells. Monoclonal T-cell receptor gene rearrangement was present in skin biopsy specimens. Although no lymphoma has been found during 2 years of follow-up treatment, the possibility of lymphoma developing out of the current smoldering stage is of concern. The clinical manifestations of lymphoproliferative disorder and chronic active EBV infection are discussed. PMID- 17910722 TI - The percentage of patients achieving PASI 75 after 1 month and remission time after climatotherapy at the Dead Sea. AB - BACKGROUND: Dead Sea climatotherapy (DSC) is a highly effective treatment for psoriasis; however, there are scanty data concerning the duration of post-therapy remission. OBJECTIVE: Assessment of the duration of remission in patients suffering from plaque-type psoriasis after a 4-week DSC. METHODS: Sixty-four patients from Germany (66% men; average age: 41.5 years) underwent a 4-week course of DSC between September 2001 and November 2002. After returning home, patients were asked to inform their dermatologist immediately when new lesions appeared. In parallel, they were called every 3 months by telephone. All patients who reported new skin lesions underwent a physical examination, including a Psoriasis Assessment Severity Index (PASI). OUTCOMES: (1) Time of recurrence of a psoriatic lesion after complete or almost complete clearance, defined as duration of remission; (2) time that elapsed until a relapse of 50% of the PASI improvement occurred, defined as duration of therapeutic effect. RESULTS: Mean PASI value before and after treatment was 31.7 and 1.42, showing a 95.5% improvement. All patients reached PASI 50, and 75.9% of them reached PASI 75 after 1 month of DSC. The median time of remission was 23.1 weeks. The median time of duration of therapeutic effect was 33.6 weeks. Statistical multivariable analysis indicated that a patient's younger age at the time of treatment was associated with a longer period of remission. CONCLUSION: Four-week DSC is an effective remittive treatment for plaque-type psoriasis. The period of remission induced compares favorably with other accepted modalities of treatment. PMID- 17910723 TI - Practical guidelines for the management of toxic epidermal necrolysis and Stevens Johnson syndrome. AB - Toxic epidermal necrolysis and Stevens-Johnson syndrome are acute life threatening dermatoses characterized by extensive sloughing and mucositis. At the University of Florida, we use practical guidelines for the management of these gravely ill patients. These can be of help to other practitioners. PMID- 17910724 TI - Etanercept for the treatment of refractory pyoderma gangrenosum: a brief series. AB - BACKGROUND: Pyoderma gangrenosum (PG) is a rare ulcerative inflammatory condition of unknown etiology. Therapy for PG involves local wound care along with topical and systemic anti-inflammatory and other immunodulatory agents. Etanercept is one such immunomodulator with activity against the inflammatory cytokine, tumor necrosis factor. OBJECTIVE: To evaluate the efficacy and safety of etanercept in the treatment of PG ulcers. METHODS: A retrospective analysis was performed on seven patients with 11 refractory PG ulcers treated with subcutaneous injections of etanercept (25-50 mg twice weekly). RESULTS: All seven patients with PG responded well to etanercept. Eight of the 11 ulcers (73%) completely healed with the mean time of (12.5 weeks), while the other three ulcers had marked reduction in size (within 8-18 weeks). Etanercept was well tolerated. No serious side effects were reported. Only one patient discontinued the drug secondary to side effects. CONCLUSION: Etanercept is an alternative treatment option for patients with refractory ulcers due to PG. PMID- 17910726 TI - Subacute prurigo-like linear IgA disease. PMID- 17910728 TI - Multiple persistent keratoacanthomas. PMID- 17910727 TI - Pimecrolimus-induced rosacea-like demodicidosis. PMID- 17910729 TI - Source localization of auditory evoked potentials after cochlear implantation. AB - Little is known about how the auditory cortex adapts to artificial input as provided by a cochlear implant (CI). We report the case of a 71-year-old profoundly deaf man, who has successfully used a unilateral CI for 4 years. Independent component analysis (ICA) of 61-channel EEG recordings could separate CI-related artifacts from auditory-evoked potentials (AEPs), even though it was the perfectly time-locked CI stimulation that caused the AEPs. AEP dipole source localization revealed contralaterally larger amplitudes in the P1-N1 range, similar to normal hearing individuals. In contrast to normal hearing individuals, the man with the CI showed a 20-ms shorter N1 latency ipsilaterally. We conclude that ICA allows the detailed study of AEPs in CI users. PMID- 17910730 TI - Neurocognitive components of the behavioral inhibition and activation systems: implications for theories of self-regulation. AB - We examined the neurocognitive correlates of the Behavioral Inhibition and Behavioral Activation Systems (BIS/BAS) in an effort to clarify ambiguities concerning interpretations of BIS as reflecting inhibition versus avoidance. We hypothesized that self-reported BIS should relate to neural mechanisms associated with conflict monitoring, whereas self-reported BAS should be associated with neural correlates of approach motivation. Consistent with these predictions, higher self-reported BIS was uniquely related to the N2 event-related potential on No-Go trials of a Go/No-Go task, linking BIS with conflict monitoring and sensitivity to No-Go cues. Higher BAS was uniquely related to greater left-sided baseline frontal cortical asymmetry associated with approach orientation. Implications for theories of self-regulation involving conflict monitoring, cognitive control, and approach/avoidance motivation are discussed. PMID- 17910732 TI - Modes of memory: early electrophysiological markers of repetition suppression and recognition enhancement predict behavioral performance. AB - Different forms of perceptual memory have opposite physiological effects. Whereas repetition priming often leads to suppression of brain responses, explicit recognition has been found to enhance brain activity. We investigated effects of repetition priming and recognition memory on early gamma-band responses. In a study phase, participants performed a visual discrimination task with task irrelevant item repetitions. Stimulus repetition suppressed early evoked gamma responses in participants with strong behavioral repetition effects. In a test phase, participants discriminated old from new items. Evoked and induced gamma activity was enhanced for old items. Effects were stronger in participants with better recognition performance. The results demonstrate a modulation of earliest stages of visual information processing by different memory systems, which is dependent on retrieval intention and predicts individual behavioral performance. PMID- 17910733 TI - The coupling of emotion and cognition in the eye: introducing the pupil old/new effect. AB - The study presented here investigated the effects of emotional valence on the memory for words by assessing both memory performance and pupillary responses during a recognition memory task. Participants had to make speeded judgments on whether a word presented in the test phase of the experiment had already been presented ("old") or not ("new"). An emotion-induced recognition bias was observed: Words with emotional content not only produced a higher amount of hits, but also elicited more false alarms than neutral words. Further, we found a distinct pupil old/new effect characterized as an elevated pupillary response to hits as opposed to correct rejections. Interestingly, this pupil old/new effect was clearly diminished for emotional words. We therefore argue that the pupil old/new effect is not only able to mirror memory retrieval processes, but also reflects modulation by an emotion-induced recognition bias. PMID- 17910731 TI - Cutaneous sensibility and peripheral nerve function in patients with unmedicated essential hypertension. AB - Sensorimotor deficits in patients with essential hypertension may be due to impaired nerve function. Cutaneous sensory thresholds, median nerve sensory and motor conduction velocities, and median nerve sensory action potential amplitudes were assessed in 30 patients with unmedicated essential hypertension and 29 normotensives. Cutaneous sensory thresholds were higher and sensory action potential amplitudes smaller in hypertensives than normotensives whereas sensory and motor nerve conduction velocities did not differ between groups. These data suggest that hypertension may reduce the number of active sensory nerve fibers without affecting myelination. Sensory action potential amplitudes were inversely related to cutaneous sensory thresholds, suggesting that subclinical axonal neuropathy of sensory afferents may help account for perceptual deficits that characterize hypertension. PMID- 17910734 TI - Effects of incentive on working memory capacity: behavioral and pupillometric data. AB - We evaluated the hypothesis that individual differences in working memory capacity are explained by variation in mental effort, persons with low capacity exerting less effort than persons with high capacity. Groups previously rated high and low in working memory capacity performed the reading span task under three levels of incentive. The effort hypothesis holds that low span subjects exert less effort during task performance than do high spans. Subjects' pupil sizes were recorded online during task performance as a measure of mental effort. Both recall performance and pupil diameter were found to be increased under incentives, but were additive with span (incentives increased performance and pupil diameter equivalently for both span groups). Contrary to the effort hypothesis, task-evoked pupillary responses indicated that if anything, low span subjects exert more effort than do high spans. PMID- 17910735 TI - The skin conductance orienting response to semantic stimuli: significance can be independent of arousal. AB - The characteristics of stimuli that elicit skin conductance responses (SCRs) have been conceptualized in varied ways, with strong emphasis on the significance or arousing quality of stimuli. Our goal was to determine whether "significance" can be shown to have an effect on SCRs independent of "arousal," using words as stimuli. Ratings of words indicated that significance is partially independent of arousal. In Study 1, SCRs from 43 participants during presentation of 20 significant, nonarousing words with a negative valence that were either depression related or potentially self-referent and 20 nonsignificant words matched on valence and arousal showed a main effect of significance. In Study 2 (N=44), significant, nonarousing words were sampled more broadly to examine the effects of self-reference and valence. Significance, rather than just negativity or self-reference, elicited SCRs independently of arousal. SCRs to significant words may reflect cognitive and attentional processes that, in turn, might prove useful for the assessment of the cognitive aspects of anxiety. PMID- 17910736 TI - Rapid picture processing: affective primes and targets. AB - In rapid serial visual presentation of pictures, an early ERP component shows enlarged negativity over occipital regions for emotional, compared to neutral, pictures. The present study examined the processing of emotional target pictures as a function of the hedonic valence of a preceding prime picture. Dense sensor ERPs were measured while subjects viewed a continuous stream of pleasant, neutral, and unpleasant pictures, presented for 335 ms each. Two main findings were observed: First, replicating previous results, emotional target pictures were associated with a larger posterior negativity, compared to neutral pictures. Second, the emotional content of the preceding prime picture affected target processing, with pleasant or unpleasant primes resulting in reduced negativity of the following picture, irrespective of its emotional valence. These findings are discussed within a framework of competition among successive pictures. PMID- 17910737 TI - Large genomic mutations within the ATM gene detected by MLPA, including a duplication of 41 kb from exon 4 to 20. AB - Mutation detection remains problematic for large genes, primarily because PCR based methodology fails to detect heterozygous deletions and any duplication. In the ATM gene only a handful of multi-exon deletions have been described to date, and this type of mutation has been considered rare. To address this issue we tested a new MLPA (Multiplex Ligation Probe Amplification) kit that covers 33 of the 66 ATM exons, using for controls two previously characterized genomic deletions in addition to three A-T patients, taken from a survey of nine, who had missing four mutations unidentified after conventional mutation screening. We identified for the first time: 1) a approximately 41 kb genomic duplication spanning exons 4-20 (c.-30_2816dup41kb)(a.k.a., ATM dup 41 kb); 2) a novel genomic deletion including exon 31, and 3) in hemizygosis a point mutation in the non-deleted exon 31. In this study we extended mutation detection to nine new Italian A-T patients, using a combined approach of haplotype analysis, DHPLC and MLPA. Overall we achieved a mutation detection rate of >97%, and can now define a spectrum of ATM mutations based on twenty-one consecutive Italian families with A T. PMID- 17910738 TI - HPA-axis activity in alcoholism: examples for a gene-environment interaction. AB - Genetic and environmental influences are both known to be causal factors in the development and maintenance of substance abuse disorders. This review aims to focus on the contributions of genetic and environmental research to the understanding of alcoholism and how gene-environment interactions result in a variety of addiction phenotypes. Gene-environment interactions have been reviewed by focusing on one of the most relevant environmental risk factors for alcoholism, stress. This is examined in more detail by reviewing the functioning of the hypothalamic-pituitary-adrenal (HPA) axis and its genetic and molecular components in this disorder. Recent evidence from animal and human studies have shown that the effects of stress on alcohol drinking are mediated by core HPA axis genes and are associated with genetic variations in those genes. The findings of the studies discussed here suggest that the collaborations of neuroscience, psychobiology and molecular genetics provide a promising framework to elucidate the exact mechanisms of gene-environment interactions as seen to convene upon the HPA axis and effect phenotypes of addiction. PMID- 17910739 TI - Changes in leptin, ghrelin, growth hormone and neuropeptide-Y after an acute model of MDMA and methamphetamine exposure in rats. AB - Club drug abuse is a growing problem in the United States. Beyond addiction and toxicity are endocrine effects which are not well characterized. Specifically, the changes in appetite following exposure to drugs of abuse are an interesting but poorly understood phenomenon. Serum hormones such as leptin, ghrelin, growth hormone (GH), and neuropeptide-Y (NP-Y) are known to affect appetite, but have not been studied extensively with drugs of abuse. In this work, we examine the effects of club drugs 3,4-methylenedioxymethamphetamine (MDMA) (ecstasy) and methamphetamine (METH) (doses of 5, 20 and 40 mg/kg) on serum concentrations of these hormones in adult male Sprague-Dawley rats 6, 12, 24 and 48 hours after drug administration. In a dose-dependent manner, MDMA was shown to cause transient significant decreases in serum leptin and GH followed by a base line recovery after 24 hours. Conversely, serum ghrelin increased and normalized after 24 hours. Interestingly, serum NP-Y showed a steady decrease in both treatment of MDMA and METH at different time points and dosages. In humans, abuse of these drugs reduces eating. As evident from these data, acute administration of METH and MDMA had significant effects on different serum hormone levels involved in appetite regulation. Future studies should be performed to see how chronic, low dose drug administration would affect hormone levels and try to answer questions about the physiological mechanisms involved in the anorexic paradigm observed in drug use. PMID- 17910740 TI - Association analysis of genes encoding the nociceptin receptor (OPRL1) and its endogenous ligand (PNOC) with alcohol or illicit drug dependence. AB - Recent studies in animal models have shown that the nociceptin system, comprising nociceptin (or OFQ/N, encoded by PNOC) and the nociceptin receptor (an opioid receptor-like protein encoded by OPRL1), may be involved in alcohol and other drug reward pathways. To determine whether the nociceptin system is associated with alcohol or illicit drug dependence in humans, we analyzed 10 single nucleotide polymorphisms (SNPs) in OPRL1 and 15 SNPs in PNOC in a sample of 1923 European Americans from 219 multiplex alcohol dependent families ascertained by the Collaborative Study on the Genetics of Alcoholism. The SNPs spanned both genes and several kb of their flanking sequences, and were in high linkage disequilibrium. Neither gene was associated with alcohol or illicit drug dependence, although two SNPs in PNOC showed marginal association with alcoholism and one with illicit drug dependence (P = 0.04-0.05). Secondary analyses suggested that two adjacent SNPs in intron 1 of OPRL1 were marginally associated with opioid dependence (P = 0.05); none of the SNPs in PNOC were associated with opioid dependence. PMID- 17910741 TI - The localized adherence pattern of an atypical enteropathogenic Escherichia coli is mediated by intimin omicron and unexpectedly promotes HeLa cell invasion. AB - Enteropathogenic Escherichia coli (EPEC) forms attaching and effacing lesions in the intestinal mucosa characterized by intimate attachment to the epithelium by means of intimin (an outer membrane adhesin encoded by eae). EPEC is subgrouped into typical (tEPEC) and atypical (aEPEC); only tEPEC carries the EAF (EPEC adherence factor) plasmid that encodes the bundle-forming pilus (BFP). Characteristically, after 3 h of incubation, tEPEC produces localized adherence (LA) (with compact microcolonies) in HeLa/HEp-2 cells by means of BFP, whereas most aEPEC form looser microcolonies. We have previously identified nine aEPEC strains displaying LA in extended (6 h) assays (LA6). In this study, we analysed the kinetics of LA6 pattern development and the role of intimin in the process. Transmission electron microscopy and confocal laser microscopy showed that the invasive process of strain 1551-2 displays a LA phenotype. An eae-defective mutant of strain 1551-2 prevented the invasion although preserving intense diffused adherence. Sequencing of eae revealed that strain 1551-2 expresses the omicron subtype of intimin. We propose that the LA phenotype of aEPEC strain 1551 2 is mediated by intimin omicron and hypothesize that this strain expresses an additional novel adhesive structure. The present study is the first to report the association of compact microcolony formation and an intense invasive ability in aEPEC. PMID- 17910742 TI - Toll-like receptor 2 plays a critical role in maintaining mucosal integrity during Citrobacter rodentium-induced colitis. AB - Inflammatory bowel diseases and infectious gastroenteritis likely occur when the integrity of intestinal barriers is disrupted allowing luminal bacterial products to cross into the intestinal mucosa, stimulating immune cells and triggering inflammation. While specific Toll-like receptors (TLR) are involved in the generation of inflammatory responses against enteric bacteria, their contributions to the maintenance of intestinal mucosal integrity are less clear. These studies investigated the role of TLR2 in a model of murine colitis induced by the bacterial pathogen Citrobacter rodentium. C. rodentium supernatants specifically activated TLR2 in vitro while infected TLR2-/- mice suffered a lethal colitis coincident with colonic mucosal ulcerations, bleeding and increased cell death but not increased pathogen burden. TLR2-/- mice suffered impaired epithelial barrier function mediated via zonula occludens (ZO)-1 in naive mice and claudin-3 in infected mice, suggesting this could underlie their susceptibility. TLR2 deficiency was also associated with impaired production of IL-6 by bone marrow-derived macrophages and infected colons cultured ex vivo. As IL-6 has antiapoptotic and epithelial repair capabilities, its reduced expression could contribute to the impaired mucosal integrity. These studies report for the first time that TLR2 plays a critical role in maintaining intestinal mucosal integrity during infection by a bacterial pathogen. PMID- 17910743 TI - Are we stalled part way through a major evolutionary transition from individual to group? PMID- 17910744 TI - Role of echocardiography in diagnosis and risk stratification in heart failure with left ventricular systolic dysfunction. AB - Heart failure (HF) is a complex clinical syndrome that can result from any structural or functional cardiac disorder that impairs the ability of the ventricle to fill with or eject blood. Echocardiography represents the "gold standard" in the assessment of LV systolic dysfunction and in the recognition of systolic heart failure, since dilatation of the LV results in alteration of intracardiac geometry and hemodynamics leading to increased morbidity and mortality. The functional mitral regurgitation is a consequence of adverse LV remodelling that occurs with a structurally normal valve and it is a marker of adverse prognosis. Diastolic dysfunction plays a major role in signs and symptoms of HF and in the risk stratification, and provides prognostic information independently in HF patients and impaired systolic function. Ultrasound lung comets are a simple echographic sign of extravascular lung water, more frequently associated with left ventricular diastolic and/or systolic dysfunction, which can integrate the clinical and pathophysiological information provided by conventional echocardiography and provide a useful information for prognostic stratification of HF patients. Contractile reserve is defined as the difference between values of an index of left ventricular contractility during peak stress and its baseline values and the presence of myocardial viability predicts a favorable outcome. A non-invasive echocardiographic method for the evaluation of force-frequency relationship has been proposed to assess the changes in contractility during stress echo. In conclusion, in HF patients, the evaluation of systolic, diastolic function and myocardial contractile reserve plays a fundamental role in the risk stratification. The highest risk is present in HF patients with a heart that is weak, big, noisy, stiff and wet. PMID- 17910745 TI - Patterns of tumor response in canine and feline cancer patients treated with electrochemotherapy: preclinical data for the standardization of this treatment in pets and humans. AB - Electrochemotherapy (ECT) is a novel anticancer therapy that is currently being evaluated in human and pet cancer patients. ECT associates the administration of an anti-tumor agent to the delivery of trains of appropriate waveforms. The increased uptake of chemotherapy leads to apoptotic death of the neoplasm thus resulting in prolonged local control and extended survival. In this paper we describe the histological features of a broad array of spontaneous tumors of companion animals receiving pulse-mediated chemotherapy. Multivariate statistical analysis of the percentage of necrosis and apoptosis in the tumors before and after ECT treatment, shows that only a high percentage of necrosis and apoptosis after the ECT treatment were significantly correlated with longer survivals of the patients (p < 0.0001 and p = 0.004, respectively). Further studies on this topic are warranted in companion animals with spontaneous tumors to identify new molecular targets for electrochemotherapy and to the develop new therapeutical protocols to be translated to humans. PMID- 17910746 TI - Dealing with sickness certification - a survey of problems and strategies among general practitioners and orthopaedic surgeons. AB - BACKGROUND: In order to get sickness benefit a sick-listed person need a medical certificate issued by a physician; in Sweden after one week of self certification. Physicians experience sick-listing tasks as problematic and conflicts may arise when patients regard themselves unable to work due to complaints that are hard to objectively verify for the physician. Most GPs and orthopaedic surgeons (OS) deal regularly with sick-listing issues in their daily practice. The aim of this study was to explore perceived problems and coping strategies related to tasks of sickness certification among general practitioners (GP) and orthopaedic surgeons (OS). METHODS: A cross-sectional study about sickness certification in two Swedish counties, with 673 participating GPs and 149 OSs, who answered a comprehensive questionnaire. Frequencies together with crude and adjusted (gender and working years) Odds ratios were calculated. RESULTS: A majority of the GPs and OSs experienced problems in sickness certification every week. To assess the patient's work ability, to handle situations when they and the patient had different opinions about the need for sickness absence, and to issue prolongation certificates when the previous was issued by another physician were reported as problematic by a majority in both groups. Both GPs and OSs prolonged sickness certifications due to waiting times in health care or at Social Insurance Office (SIO). To handle experienced problems they used different strategies; OSs issued sickness certificates without personal appointment more often than the GPs, who on the other hand reported having contact with SIO more often than the OSs. A higher rate of GPs experienced support from management and had a common strategy for handling sickness certification at the clinic than the OSs. CONCLUSION: Most GPs and OSs handled sickness certification weekly and reported a variety of problems in relation to this task, generally GPs to a higher extent, and they used different coping strategies to handle the problems. PMID- 17910748 TI - Prevalence of childhood and early adolescence mental disorders among children attending primary health care centers in Mosul, Iraq: a cross-sectional study. AB - BACKGROUND: Children and adolescents are more vulnerable to the affects of war and violence than adults. At the time of initiation of this study, nothing was known about the prevalence of childhood and early adolescence mental disorders. The aim of the present study is to measure the point prevalence of mental disorders among children of 1-15 years age in the city of Mosul, Iraq. METHODS: A cross-sectional study design was adopted. Four primary health care centers were chosen consecutively as a study setting. The subjects of the present study were mothers who came to the primary health care center for vaccination of their children. The chosen mothers were included by systematic sampling randomization. All children (aged 1-15) that each mother had were considered in the interview and examination. RESULTS: Out of 3079 children assessed, 1152 have childhood mental disorders, giving a point prevalence of 37.4%, with a male to female ratio of to 1.22:1. The top 10 disorders among the examined children are post-traumatic stress disorder (10.5%), enuresis (6%), separation anxiety disorder (4.3%), specific phobia (3.3%) stuttering and refusal to attend school (3.2% each), learning and conduct disorders (2.5% each), stereotypic movement (2.3%) and feeding disorder in infancy or early childhood (2.0%). Overall, the highest prevalence of mental disorders was among children 10-15 years old (49.2%) while the lowest was among 1-5 year olds (29.1%). Boys are more affected than girls (40.2% and 33.2%, respectively). CONCLUSION: Childhood mental disorders are a common condition highly prevalent amongst the children and early adolescents in Mosul. Data from the present study mirrors the size of the problem in local community. Several points deserve attention, the most important of which include giving care at the community level, educating the public on mental health, involving communities and families, monitoring community mental health indicators, and providing treatment at primary health care level. PMID- 17910747 TI - Telmisartan/hydrochlorothiazide versus valsartan/hydrochlorothiazide in obese hypertensive patients with type 2 diabetes: the SMOOTH study. AB - BACKGROUND: The Study of Micardis (telmisartan) in Overweight/Obese patients with Type 2 diabetes and Hypertension (SMOOTH) compared hydrochlorothiazide (HCTZ) plus telmisartan or valsartan fixed-dose combination therapies on early morning blood pressure (BP), using ambulatory BP monitoring (ABPM). METHODS: SMOOTH was a prospective, randomized, open-label, blinded-endpoint, multicentre trial. After a 2- to 4-week, single-blind, placebo run-in period, patients received once-daily telmisartan 80 mg or valsartan 160 mg for 4 weeks, with add-on HCTZ 12.5 mg for 6 weeks (T/HCTZ or V/HCTZ, respectively). At baseline and week 10, ambulatory blood pressure (ABP) was measured every 20 min and hourly means were calculated. The primary endpoint was change from baseline in mean ambulatory systolic and diastolic blood pressure (SBP; DBP) during the last 6 hours of the 24-hour dosing interval. RESULTS: In total, 840 patients were randomized. At week 10, T/HCTZ provided significantly greater reductions versus V/HCTZ in the last 6 hours mean ABP (differences in favour of T/HCTZ: SBP 3.9 mm Hg, p < 0.0001; DBP 2.0 mm Hg, p = 0.0007). T/HCTZ also produced significantly greater reductions than V/HCTZ in 24-hour mean ABP (differences in favour of T/HCTZ: SBP 3.0 mm Hg, p = 0.0002; DBP 1.6 mm Hg, p = 0.0006) and during the morning, daytime and night-time periods (p < 0.003). Both treatments were well tolerated. CONCLUSION: In high-risk, overweight/obese patients with hypertension and type 2 diabetes, T/HCTZ provides significantly greater BP lowering versus V/HCTZ throughout the 24-hour dosing interval, particularly during the hazardous early morning hours. PMID- 17910749 TI - Metastatic malignant melanoma in bone marrow with occult primary site--a case report with review of literature. AB - BACKGROUND: Metastases of malignant melanoma to the bone marrow are very rare. A few case reports are published in the literature with a known primary site. CASE PRESENTATION: Herein we present a case of metastatic malignant melanoma in bone marrow with occult primary site in a 22-year-old-male. Diagnosis was confirmed by morphology and immunohistochemistry. A pertinent review of literature is also presented by using relevant articles indexed in PubMed (National Library of Medicine) database. The search was based on the following terms: metastasis or metastases, malignant melanoma and bone marrow. CONCLUSION: In this report we discuss a rare case of metastatic malignant melanoma to the bone marrow with an unknown primary. Clinicians must be aware of the varied clinical manifestations of disseminated malignant melanoma even if the primary site is not evident. PMID- 17910750 TI - The mental health disaster in conflict settings: can scientific research help? PMID- 17910751 TI - Mesenteric panniculitis of the sigmoid colon: a case report and review of the literature. AB - INTRODUCTION: Mesenteric panniculitis of the sigmoid colon is a rare occurrence in surgical practice. The aim of this article is to present a case of mesenteric panniculitis of the sigmoid colon and a short review of the literature. CASE PRESENTATION: We reviewed the hospital record of a 63-year-old man who presented with a palpable mass in the left abdomen and clinical signs of a partial bowel obstruction. The pre-operative impression was a possible cancer of the sigmoid colon. A laparotomy was performed through a midline incision. The mesentery was found to be markedly thickened, constricted and puckered. The normal architecture of the adipose tissue had been lost and replaced with an irregular nodular mass. The microscopic pathologic sections demonstrated a chronic reactive inflammatory process with an exuberant proliferation of fibroblasts and fibrocytes. The adipose tissue contained scattered areas of steatonecrosis with foci of lipid laden macrophages, lymphocytes and plasma cells. The sigmoid colon and its mesocolon were resected. The postoperative course was uneventful and the patient was discharged in good condition, and followed up for the next two years. CONCLUSION: Mesenteric panniculitis of sigmoid is an extremely rare entity of unknown origin in which the normal architecture of the mesentery is replaced by fibrosis, necrosis and calcification. On gross examination the alterations may be mistaken for a neoplastic process. A frozen section may be necessary for confirmation of the diagnosis. When the advanced inflammatory changes became irreversible and bowel obstruction occurs, resection may be indicated. PMID- 17910752 TI - A convenient allylsilane-N-acyliminium route toward indolizidine and quinolizidine alkaloids. AB - This review relates all the results that we obtained in the field of the total synthesis of indolizidine and quinolizidine alkaloids using a strategy of the addition of an allylsilane on an N-acyliminium ion. In this paper, we describe the synthesis of racemic indolizidine 167B and chiral indolizidines: (-) indolizidines 167B, 195B, 223AB, (+)-monomorine, (-)-(3R,5S,8aS)-3-butyl-5 propylindolizidine and (-)-dendroprimine. Next, we relate the synthesis that we have developed in the quinolizidines field: (+/-)-myrtine and epimyrtine, (+/-) lasubines I and II and chiral quinolizidines: (+)-myrtine, (-)-epimyrtine, (-) lasubines I and II and (+)-subcosine II. PMID- 17910753 TI - Altered expression pattern of integrin alphavbeta3 correlates with actin cytoskeleton in primary cultures of human breast cancer. AB - BACKGROUND: Integrins are transmembrane adhesion receptors that provide the physical link between the actin cytoskeleton and the extracellular matrix. It has been well established that integrins play a major role in various cancer stages, such as tumor growth, progression, invasion and metastasis. In breast cancer, integrin alphavbeta3 has been associated with high malignant potential in cancer cells, signaling the onset of widespread metastasis. Many preclinical breast cancer studies are based on established cell lines, which may not represent the cell behavior and phenotype of the primary tumor of origin, due to undergone genotypic and phenotypic changes. In the present study, short-term primary breast cancer cell cultures were developed. Integrin alphavbeta3 localization was studied in correlation with F-actin cytoskeleton by means of immunofluorescence and immunogold ultrastructural localization. Integrin fluorescence intensities were semi-quantitatively assessed by means of computerized image analysis, while integrin and actin expression was evaluated by Western immunoblotting. RESULTS: In the primary breast cancer epithelial cells integrin alphavbeta3 immunofluorescence was observed in the marginal cytoplasmic area, whereas in the primary normal breast epithelial cells it was observed in the main cell body, i.e. in the ventrally located perinuclear area. In the former, F-actin cytoskeleton appeared well-formed, consisting of numerous and thicker stress fibers, compared to normal epithelial cells. Furthermore, electron microscopy showed increased integrin alphavbeta3 immunogold localization in epithelial breast cancer cells over the area of stress fibers at the basal cell surface. These findings were verified with Western immunoblotting by the higher expression of integrin beta3 subunit and actin in primary breast cancer cells, revealing their reciprocal relation, in response to the higher motility requirements, determined by the malignant potential of the breast cancer cells. CONCLUSION: A model system of primary breast cancer cell cultures was developed, in an effort to maintain the closest resembling environment to the tumor of origin. Using the above system model as an experimental tool the study of breast tumor cell behavior is possible concerning the adhesion capacity and the migrating potential of these cells, as defined by the integrin alphavbeta3 distribution in correlation with F-actin cytoskeleton. PMID- 17910754 TI - Activities contributing to energy expenditure among Guatemalan adults. AB - BACKGROUND: Guatemala has experienced a substantial increase in overweight and obesity in recent years, yet physical activity patterns and consequent energy expenditure are largely unexplored in this population. METHODS: To describe overall physical activity levels (PAL) and activities contributing to daily energy expenditure, we analyzed time spent in daily activities as reported by 985 women and 819 men, living in rural and urban areas of Guatemala in 2002-04. RESULTS: Physical activity levels recommended to prevent obesity (PAL > or = 1.70) differed by residence/occupation among men (agricultural-rural: 77%; nonagricultural-rural: 36%; urban: 24%; P < 0.01), but not women (rural: 2%; urban: 3%; P = 0.5). Median energy expenditure was higher among agricultural rural men (44 MET*h/d; MET = metabolic equivalent) compared to nonagricultural rural (37 MET*h/d) and urban men (35 MET*h/d; P < 0.01); energy expenditure was slightly lower among rural compared to urban women (34 MET*h/d vs. 35 MET*h/d; P < 0.01). Occupation was the largest contributor to energy expenditure (19-24 MET*h/d); among women and nonagricultural-rural and urban men this was primarily of a light intensity. Energy expenditure in sedentary activities ranged from 2 MET*h/d among rural women to 6 MET*h/d among agricultural-rural men. Any sports/exercise time was reported by 35% and 5% of men and women, respectively. Nevertheless, the majority of participants believed they were significantly active to stay healthy. CONCLUSION: Overall, energy expenditure was low in the population not dedicated to agricultural occupations; an increased focus on active leisure-time behaviors may be needed to counterbalance reductions in energy expenditure consequent to sedentarization of primary occupations. PMID- 17910755 TI - Treating anemia of chronic kidney disease in the primary care setting: cardiovascular outcomes and management recommendations. AB - Anemia is an underrecognized but characteristic feature of chronic kidney disease (CKD), associated with significant cardiovascular morbidity, hospitalization, and mortality. Since their inception nearly two decades ago, erythropoiesis-stimulating agents (ESAs) have revolutionized the care of patients with renal anemia, and their use has been associated with improved quality of life and reduced hospitalizations, inpatient costs, and mortality. Hemoglobin targets >/=13 g/dL have been linked with adverse events in recent randomized trials, raising concerns over the proper hemoglobin range for ESA treatment. This review appraises observational and randomized studies of the outcomes of erythropoietic treatment and offers recommendations for managing renal anemia in the primary care setting. PMID- 17910756 TI - An investigation of factors associated with psychiatric hospital admission despite the presence of crisis resolution teams. AB - BACKGROUND: Crisis resolution teams (CRTs) provide a community alternative to psychiatric hospital admission for patients presenting in crisis. Little is known about the characteristics of patients admitted despite the availability of such teams. METHODS: Data were drawn from three investigations of the outcomes of CRTs in inner London. A literature review was used to identify candidate explanatory variables that may be associated with admission despite the availability of intensive home treatment. The main outcome variable was admission to hospital within 8 weeks of the initial crisis. Associations between this outcome and the candidate explanatory variables were tested using first univariate and then multivariate analysis. RESULTS: Patients who were uncooperative with initial assessment (OR 10.25 95% CI-4.20-24.97), at risk of self-neglect (OR 2.93 1.42 6.05), had a history of compulsory admission (OR 2.64 1.07-6.55), assessed outside usual office hours (OR 2.34 1.11-4.94) and/or were assessed in hospital casualty departments (OR 3.12 1.55-6.26), were more likely to be admitted. Other than age, no socio-demographic features or diagnostic variables were significantly associated with risk of admission. CONCLUSION: With the introduction of CRTs, inpatient wards face a significant challenge, as patients who cooperate little with treatment, neglect themselves, or have previously been compulsorily detained are especially likely to be admitted. The increased risk of admission associated with casualty department assessment may be remediable. PMID- 17910757 TI - Production of L-carnitine by secondary metabolism of bacteria. AB - The increasing commercial demand for L-carnitine has led to a multiplication of efforts to improve its production with bacteria. The use of different cell environments, such as growing, resting, permeabilized, dried, osmotically stressed, freely suspended and immobilized cells, to maintain enzymes sufficiently active for L-carnitine production is discussed in the text. The different cell states of enterobacteria, such as Escherichia coli and Proteus sp., which can be used to produce L-carnitine from crotonobetaine or D-carnitine as substrate, are analyzed. Moreover, the combined application of both bioprocess and metabolic engineering has allowed a deeper understanding of the main factors controlling the production process, such as energy depletion and the alteration of the acetyl-CoA/CoA ratio which are coupled to the end of the biotransformation. Furthermore, the profiles of key central metabolic activities such as the TCA cycle, the glyoxylate shunt and the acetate metabolism are seen to be closely interrelated and affect the biotransformation efficiency. Although genetically modified strains have been obtained, new strain improvement strategies are still needed, especially in Escherichia coli as a model organism for molecular biology studies. This review aims to summarize and update the state of the art in L-carnitine production using E. coli and Proteus sp, emphasizing the importance of proper reactor design and operation strategies, together with metabolic engineering aspects and the need for feed-back between wet and in silico work to optimize this biotransformation. PMID- 17910758 TI - Ptf1a triggers GABAergic neuronal cell fates in the retina. AB - BACKGROUND: In recent years, considerable knowledge has been gained on the molecular mechanisms underlying retinal cell fate specification. However, hitherto studies focused primarily on the six major retinal cell classes (five types of neurons of one type of glial cell), and paid little attention to the specification of different neuronal subtypes within the same cell class. In particular, the molecular machinery governing the specification of the two most abundant neurotransmitter phenotypes in the retina, GABAergic and glutamatergic, is largely unknown. In the spinal cord and cerebellum, the transcription factor Ptf1a is essential for GABAergic neuron production. In the mouse retina, Ptf1a has been shown to be involved in horizontal and most amacrine neurons differentiation. RESULTS: In this study, we examined the distribution of neurotransmitter subtypes following Ptf1a gain and loss of function in the Xenopus retina. We found cell-autonomous dramatic switches between GABAergic and glutamatergic neuron production, concomitant with profound defects in the genesis of amacrine and horizontal cells, which are mainly GABAergic. Therefore, we investigated whether Ptf1a promotes the fate of these two cell types or acts directly as a GABAergic subtype determination factor. In ectodermal explant assays, Ptf1a was found to be a potent inducer of the GABAergic subtype. Moreover, clonal analysis in the retina revealed that Ptf1a overexpression leads to an increased ratio of GABAergic subtypes among the whole amacrine and horizontal cell population, highlighting its instructive capacity to promote this specific subtype of inhibitory neurons. Finally, we also found that within bipolar cells, which are typically glutamatergic interneurons, Ptf1a is able to trigger a GABAergic fate. CONCLUSION: Altogether, our results reveal for the first time in the retina a major player in the GABAergic versus glutamatergic cell specification genetic pathway. PMID- 17910759 TI - Gene expression profiling and histopathological characterization of triple negative/basal-like breast carcinomas. AB - INTRODUCTION: Breast cancer is a heterogeneous group of tumors, and can be subdivided on the basis of histopathological features, genetic alterations and gene-expression profiles. One well-defined subtype of breast cancer is characterized by a lack of HER2 gene amplification and estrogen and progesterone receptor expression ('triple-negative tumors'). We examined the histopathological and gene-expression profile of triple-negative tumors to define subgroups with specific characteristics, including risk of developing distant metastases. METHODS: 97 triple-negative tumors were selected from the fresh-frozen tissue bank of the Netherlands Cancer Institute, and gene-expression profiles were generated using 35K oligonucleotide microarrays. In addition, histopathological and immunohistochemical characterization was performed, and the findings were associated to clinical features. RESULTS: All triple-negative tumors were classified as basal-like tumors on the basis of their overall gene-expression profile. Hierarchical cluster analysis revealed five distinct subgroups of triple negative breast cancers. Multivariable analysis showed that a large amount of lymphocytic infiltrate (HR = 0.30, 95% CI 0.09-0.96) and absence of central fibrosis in the tumors (HR = 0.14, 95% CI 0.03-0.62) were associated with distant metastasis-free survival. CONCLUSION: Triple-negative tumors are synonymous with basal-like tumors, and can be identified by immunohistochemistry. Based on gene expression profiling, basal-like tumors are still heterogeneous and can be subdivided into at least five distinct subgroups. The development of distant metastasis in basal-like tumors is associated with the presence of central fibrosis and a small amount of lymphocytic infiltrate. PMID- 17910761 TI - Comparative genomic analysis of Tropheryma whipplei strains reveals that diversity among clinical isolates is mainly related to the WiSP proteins. AB - BACKGROUND: The aim of this study was to analyze the genomic diversity of several Tropheryma whipplei strains by microarray-based comparative genomic hybridization. Fifteen clinical isolates originating from biopsy samples recovered from different countries were compared with the T. whipplei Twist strain. For each isolate, the genes were defined as either present or absent/divergent using the GACK analysis software. Genomic changes were then further characterized by PCR and sequencing. RESULTS: The results revealed a limited genetic variation among the T. whipplei isolates, with at most 2.24% of the probes exhibiting differential hybridization against the Twist strain. The main variation was found in genes encoding the WiSP membrane protein family. This work also demonstrated a 19.2 kb-pair deletion within the T. whipplei DIG15 strain. This deletion occurs in the same region as the previously described large genomic rearrangement between Twist and TW08/27. Thus, this can be considered as a major hot-spot for intra-specific T. whipplei differentiation. Analysis of this deleted region confirmed the role of WND domains in generating T. whipplei diversity. CONCLUSION: This work provides the first comprehensive genomic comparison of several T. whipplei isolates. It reveals that clinical isolates originating from various geographic and biological sources exhibit a high conservation rate, indicating that T. whipplei rarely interacts with exogenous DNA. Remarkably, frequent inter-strain variations were dicovered that affected members of the WiSP family. PMID- 17910760 TI - Detection, characterization and regulation of antisense transcripts in HIV-1. AB - BACKGROUND: We and others have recently demonstrated that the human retrovirus HTLV-I was producing a spliced antisense transcript, which led to the synthesis of the HBZ protein. The objective of the present study was to demonstrate the existence of antisense transcription in HIV-1 and to provide a better characterization of the transcript and its regulation. RESULTS: Initial experiments conducted by standard RT-PCR analysis in latently infected J1.1 cell line and pNL4.3-transfected 293T cells confirmed the existence of antisense transcription in HIV-1. A more adapted RT-PCR protocol with limited RT-PCR artefacts also led to a successful detection of antisense transcripts in several infected cell lines. RACE analyses demonstrated the existence of several transcription initiation sites mapping near the 5' border of the 3'LTR (in the antisense strand). Interestingly, a new polyA signal was identified on the antisense strand and harboured the polyA signal consensus sequence. Transfection experiments in 293T and Jurkat cells with an antisense luciferase-expressing NL4.3 proviral DNA showed luciferase reporter gene expression, which was further induced by various T-cell activators. In addition, the viral Tat protein was found to be a positive modulator of antisense transcription by transient and stable transfections of this proviral DNA construct. RT-PCR analyses in 293T cells stably transfected with a pNL4.3-derived construct further confirmed these results. Infection of 293T, Jurkat, SupT1, U937 and CEMT4 cells with pseudotyped virions produced from the antisense luciferase-expressing NL4.3 DNA clone led to the production of an AZT-sensitive luciferase signal, which was however less pronounced than the signal from NL4.3Luc-infected cells. CONCLUSION: These results demonstrate for the first time that antisense transcription exists in HIV 1 in the context of infection. Possible translation of the predicted antisense ORF in this transcript should thus be re-examined. PMID- 17910762 TI - Validation of ICD-9-CM/ICD-10 coding algorithms for the identification of patients with acetaminophen overdose and hepatotoxicity using administrative data. AB - BACKGROUND: Acetaminophen overdose is the most common cause of acute liver failure (ALF). Our objective was to develop coding algorithms using administrative data for identifying patients with acetaminophen overdose and hepatic complications. METHODS: Patients hospitalized for acetaminophen overdose were identified using population-based administrative data (1995-2004). Coding algorithms for acetaminophen overdose, hepatotoxicity (alanine aminotransferase >1,000 U/L) and ALF (encephalopathy and international normalized ratio >1.5) were derived using chart abstraction data as the reference and logistic regression analyses. RESULTS: Of 1,776 potential acetaminophen overdose cases, the charts of 181 patients were reviewed; 139 (77%) had confirmed acetaminophen overdose. An algorithm including codes 965.4 (ICD-9-CM) and T39.1 (ICD-10) was highly accurate (sensitivity 90% [95% confidence interval 84-94%], specificity 83% [69-93%], positive predictive value 95% [89-98%], negative predictive value 71% [57-83%], c statistic 0.87 [0.80-0.93]). Algorithms for hepatotoxicity (including codes for hepatic necrosis, toxic hepatitis and encephalopathy) and ALF (hepatic necrosis and encephalopathy) were also highly predictive (c-statistics = 0.88). The accuracy of the algorithms was not affected by age, gender, or ICD coding system, but the acetaminophen overdose algorithm varied between hospitals (c-statistics 0.84-0.98; P = 0.003). CONCLUSION: Administrative databases can be used to identify patients with acetaminophen overdose and hepatic complications. If externally validated, these algorithms will facilitate investigations of the epidemiology and outcomes of acetaminophen overdose. PMID- 17910764 TI - Obituary: arun fotedar. PMID- 17910763 TI - Cadmium triggers an integrated reprogramming of the metabolism of Synechocystis PCC6803, under the control of the Slr1738 regulator. AB - BACKGROUND: Cadmium is a persistent pollutant that threatens most biological organisms, including cyanobacteria that support a large part of the biosphere. Using a multifaceted approach, we have investigated the global responses to Cd and other relevant stresses (H2O2 and Fe) in the model cyanobacterium Synechocystis PCC6803. RESULTS: We found that cells respond to the Cd stress in a two main temporal phases process. In the "early" phase cells mainly limit Cd entry through the negative and positive regulation of numerous genes operating in metal uptake and export, respectively. As time proceeds, the number of responsive genes increases. In this "massive" phase, Cd downregulates most genes operating in (i) photosynthesis (PS) that normally provides ATP and NADPH; (ii) assimilation of carbon, nitrogen and sulfur that requires ATP and NAD(P)H; and (iii) translation machinery, a major consumer of ATP and nutrients. Simultaneously, many genes are upregulated, such as those involved in Fe acquisition, stress tolerance, and protein degradation (crucial to nutrients recycling). The most striking common effect of Cd and H2O2 is the disturbance of both light tolerance and Fe homeostasis, which appeared to be interdependent. Our results indicate that cells challenged with H2O2 or Cd use different strategies for the same purpose of supplying Fe atoms to Fe-requiring metalloenzymes and the SUF machinery, which synthesizes or repairs Fe-S centers. Cd-stressed cells preferentially breakdown their Fe-rich PS machinery, whereas H2O2-challenged cells preferentially accelerate the intake of Fe atoms from the medium. CONCLUSION: We view the responses to Cd as an integrated "Yin Yang" reprogramming of the whole metabolism, we found to be controlled by the Slr1738 regulator. As the Yin process, the ATP- and nutrients-sparing downregulation of anabolism limits the poisoning incorporation of Cd into metalloenzymes. As the compensatory Yang process, the PS breakdown liberates nutrient assimilates for the synthesis of Cd-tolerance proteins, among which we found the Slr0946 arsenate reductase enzyme. PMID- 17910765 TI - A pandemic strain of calicivirus threatens rabbit industries in the Americas. AB - Rabbit Hemorrhagic Disease (RHD) is a severe acute viral disease specifically affecting the European rabbit Oryctolagus cuniculus. As the European rabbit is the predominant species of domestic rabbit throughout the world, RHD contributes towards significant losses to rabbit farming industries and endangers wild populations of rabbits in Europe and other predatory animals in Europe that depend upon rabbits as a food source. Rabbit Hemorrhagic Disease virus (RHDV) - a Lagovirus belonging to the family Caliciviridae is the etiological agent of RHD. Typically, RHD presents with sudden death in 70% to 95% of infected animals. There have been four separate incursions of RHDV in the USA, the most recent of which occurred in the state of Indiana in June of 2005. Animal inoculation studies confirmed the pathogenicity of the Indiana 2005 isolate, which caused acute death and pathological changes characterized by acute diffuse severe liver necrosis and pulmonary hemorrhages. Complete viral genome sequences of all USA outbreak isolates were determined and comparative genomics revealed that each outbreak was the result of a separate introduction of virus rather than from a single virus lineage. All of the USA isolates clustered with RHDV genomes from China, and phylogenetic analysis of the major capsid protein (VP60) revealed that they were related to a pandemic antigenic variant strain known as RHDVa. Rapid spread of the RHDVa pandemic suggests a selective advantage for this new subtype. Given its rapid spread, pathogenic nature, and potential to further evolve, possibly broadening its host range to include other genera native to the Americas, RHDVa should be regarded as a threat. PMID- 17910766 TI - BNDB - the Biochemical Network Database. AB - BACKGROUND: Technological advances in high-throughput techniques and efficient data acquisition methods have resulted in a massive amount of life science data. The data is stored in numerous databases that have been established over the last decades and are essential resources for scientists nowadays. However, the diversity of the databases and the underlying data models make it difficult to combine this information for solving complex problems in systems biology. Currently, researchers typically have to browse several, often highly focused, databases to obtain the required information. Hence, there is a pressing need for more efficient systems for integrating, analyzing, and interpreting these data. The standardization and virtual consolidation of the databases is a major challenge resulting in a unified access to a variety of data sources. DESCRIPTION: We present the Biochemical Network Database (BNDB), a powerful relational database platform, allowing a complete semantic integration of an extensive collection of external databases. BNDB is built upon a comprehensive and extensible object model called BioCore, which is powerful enough to model most known biochemical processes and at the same time easily extensible to be adapted to new biological concepts. Besides a web interface for the search and curation of the data, a Java-based viewer (BiNA) provides a powerful platform independent visualization and navigation of the data. BiNA uses sophisticated graph layout algorithms for an interactive visualization and navigation of BNDB. CONCLUSION: BNDB allows a simple, unified access to a variety of external data sources. Its tight integration with the biochemical network library BN++ offers the possibility for import, integration, analysis, and visualization of the data. BNDB is freely accessible at http://www.bndb.org. PMID- 17910767 TI - Assessment of algorithms for high throughput detection of genomic copy number variation in oligonucleotide microarray data. AB - BACKGROUND: Genomic deletions and duplications are important in the pathogenesis of diseases, such as cancer and mental retardation, and have recently been shown to occur frequently in unaffected individuals as polymorphisms. Affymetrix GeneChip whole genome sampling analysis (WGSA) combined with 100 K single nucleotide polymorphism (SNP) genotyping arrays is one of several microarray based approaches that are now being used to detect such structural genomic changes. The popularity of this technology and its associated open source data format have resulted in the development of an increasing number of software packages for the analysis of copy number changes using these SNP arrays. RESULTS: We evaluated four publicly available software packages for high throughput copy number analysis using synthetic and empirical 100 K SNP array data sets, the latter obtained from 107 mental retardation (MR) patients and their unaffected parents and siblings. We evaluated the software with regards to overall suitability for high-throughput 100 K SNP array data analysis, as well as effectiveness of normalization, scaling with various reference sets and feature extraction, as well as true and false positive rates of genomic copy number variant (CNV) detection. CONCLUSION: We observed considerable variation among the numbers and types of candidate CNVs detected by different analysis approaches, and found that multiple programs were needed to find all real aberrations in our test set. The frequency of false positive deletions was substantial, but could be greatly reduced by using the SNP genotype information to confirm loss of heterozygosity. PMID- 17910768 TI - SAMMD: Staphylococcus aureus microarray meta-database. AB - BACKGROUND: Staphylococcus aureus is an important human pathogen, causing a wide variety of diseases ranging from superficial skin infections to severe life threatening infections. S. aureus is one of the leading causes of nosocomial infections. Its ability to resist multiple antibiotics poses a growing public health problem. In order to understand the mechanism of pathogenesis of S. aureus, several global expression profiles have been developed. These transcriptional profiles included regulatory mutants of S. aureus and growth of wild type under different growth conditions. The abundance of these profiles has generated a large amount of data without a uniform annotation system to comprehensively examine them. We report the development of the Staphylococcus aureus Microarray meta-database (SAMMD) which includes data from all the published transcriptional profiles. SAMMD is a web-accessible database that helps users to perform a variety of analysis against and within the existing transcriptional profiles. DESCRIPTION: SAMMD is a relational database that uses MySQL as the back end and PHP/JavaScript/DHTML as the front end. The database is normalized and consists of five tables, which holds information about gene annotations, regulated gene lists, experimental details, references, and other details. SAMMD data is collected from the peer-reviewed published articles. Data extraction and conversion was done using perl scripts while data entry was done through phpMyAdmin tool. The database is accessible via a web interface that contains several features such as a simple search by ORF ID, gene name, gene product name, advanced search using gene lists, comparing among datasets, browsing, downloading, statistics, and help. The database is licensed under General Public License (GPL). CONCLUSION: SAMMD is hosted and available at http://www.bioinformatics.org/sammd/. Currently there are over 9500 entries for regulated genes, from 67 microarray experiments. SAMMD will help staphylococcal scientists to analyze their expression data and understand it at global level. It will also allow scientists to compare and contrast their transcriptome to that of the other published transcriptomes. PMID- 17910769 TI - Adherence to colorectal cancer screening guidelines in Canada. AB - BACKGROUND: To identify correlates of adherence to colorectal cancer (CRC) screening guidelines in average-risk Canadians. METHODS: 2003 Canadian Community Health Survey Cycle 2.1 respondents who were at least 50 years old, without past or present CRC and living in Ontario, Newfoundland, Saskatchewan, and British Columbia were included. Outcomes, defined according to current CRC screening guidelines, included adherence to: i) fecal occult blood test (FOBT) (in prior 2 years), ii) endoscopy (colonoscopy/sigmoidoscopy) (prior 10 years), and iii) adherence to CRC screening guidelines, defined as either (i) or (ii). Generalized estimating equations regression was employed to identify correlates of the study outcomes. RESULTS: Of the 17,498 respondents, 70% were non-adherent CRC screening to guidelines. Specifically, 85% and 79% were non-adherent to FOBT and endoscopy, respectively. Correlates for all outcomes were: having a regular physician (OR = (i) 2.68; (ii) 1.91; (iii) 2.39), getting a flu shot (OR = (i) 1.59; (ii) 1.51; (iii) 1.55), and having a chronic condition (OR = (i) 1.32; (ii) 1.48; (iii) 1.43). Greater physical activity, higher consumption of fruits and vegetables and smoking cessation were each associated with at least 1 outcome. Self-perceived stress was modestly associated with increased odds of adherence to endoscopy and to CRC screening guidelines (OR = (ii) 1.07; (iii) 1.06, respectively). CONCLUSION: Healthy lifestyle behaviors and factors that motivate people to seek health care were associated with adherence, implying that invitations for CRC screening should come from sources that are independent of physicians, such as the government, in order to reduce disparities in CRC screening. PMID- 17910771 TI - Misleading pustular plaques of the lower limbs during Crohn's disease: two case reports. AB - BACKGROUND: Extraintestinal manifestations of Crohn's disease may involve the skin, the eyes, the genital mucosa, and the joints. Dermatoses associated with Crohn's disease include neutrophilic dermatoses, erythema nodosum, granulomatous dermatitis, blistering dermatoses, and non-specific skin manifestations. Cutaneous Crohn's disease is characterized by skin non-caseating epithelioid granulomatas with giant cells, remote from the gastrointestinal tract. We report herein two new cases. OBSERVATIONS: On both patients, differential diagnosis of neutrophilic dermatoses and infectious disease were evoked, and antimicrobial agents were introduced in one of them. Given the atypical presentation, the final diagnosis of cutaneous Crohn's disease could only be made with histological examination. In patient 1, the plaques decreased in size and infiltration by more than 75% after 3 weeks of treatment with bethametasone dipropionate 0.05% cream. In patient 2, the plaques decreased by more than 50% after 6 weeks of treatment with prednisolone (45 mg/day) and azathioprine (100 mg/day). DISCUSSION: Cutaneous Crohn's disease may present as dusky, erythematous, infiltrated, and ulcerated plaques and nodules. Female-to-male sex ratio is about 2, and the mean age at onset is 35. Recurrently, the hypothesis of a skin mycobacterial or fungal infection greatly delays proper treatment. Rarity of cutaneous Crohn's disease hampers therapeutic assessment in controlled trials. Thus, available literature is limited to case reports and sparse small series, with contradictory results. These reports are subject to publication bias, and no definite evidence-based recommendations can be made on the most adequate therapeutic strategy. PMID- 17910770 TI - Antibody-protein interactions: benchmark datasets and prediction tools evaluation. AB - BACKGROUND: The ability to predict antibody binding sites (aka antigenic determinants or B-cell epitopes) for a given protein is a precursor to new vaccine design and diagnostics. Among the various methods of B-cell epitope identification X-ray crystallography is one of the most reliable methods. Using these experimental data computational methods exist for B-cell epitope prediction. As the number of structures of antibody-protein complexes grows, further interest in prediction methods using 3D structure is anticipated. This work aims to establish a benchmark for 3D structure-based epitope prediction methods. RESULTS: Two B-cell epitope benchmark datasets inferred from the 3D structures of antibody-protein complexes were defined. The first is a dataset of 62 representative 3D structures of protein antigens with inferred structural epitopes. The second is a dataset of 82 structures of antibody-protein complexes containing different structural epitopes. Using these datasets, eight web-servers developed for antibody and protein binding sites prediction have been evaluated. In no method did performance exceed a 40% precision and 46% recall. The values of the area under the receiver operating characteristic curve for the evaluated methods were about 0.6 for ConSurf, DiscoTope, and PPI-PRED methods and above 0.65 but not exceeding 0.70 for protein-protein docking methods when the best of the top ten models for the bound docking were considered; the remaining methods performed close to random. The benchmark datasets are included as a supplement to this paper. CONCLUSION: It may be possible to improve epitope prediction methods through training on datasets which include only immune epitopes and through utilizing more features characterizing epitopes, for example, the evolutionary conservation score. Notwithstanding, overall poor performance may reflect the generality of antigenicity and hence the inability to decipher B-cell epitopes as an intrinsic feature of the protein. It is an open question as to whether ultimately discriminatory features can be found. PMID- 17910772 TI - Co-expression of adjacent genes in yeast cannot be simply attributed to shared regulatory system. AB - BACKGROUND: Adjacent gene pairs in the yeast genome have a tendency to express concurrently. Sharing of regulatory elements within the intergenic region of those adjacent gene pairs was often considered the major mechanism responsible for such co-expression. However, it is still in debate to what extent that common transcription factors (TFs) contribute to the co-expression of adjacent genes. In order to resolve the evolutionary aspect of this issue, we investigated the conservation of adjacent pairs in five yeast species. By using the information for TF binding sites in promoter regions available from the MYBS database http://cg1.iis.sinica.edu.tw/~mybs/, the ratios of TF-sharing pairs among all the adjacent pairs in yeast genomes were analyzed. The levels of co-expression in different adjacent patterns were also compared. RESULTS: Our analyses showed that the proportion of adjacent pairs conserved in five yeast species is relatively low compared to that in the mammalian lineage. The proportion was also low for adjacent gene pairs with shared TFs. Particularly, the statistical analysis suggested that co-expression of adjacent gene pairs was not noticeably associated with the sharing of TFs in these pairs. We further proposed a case of the PAC (polymerase A and C) and RRPE (rRNA processing element) motifs which co-regulate divergent/bidirectional pairs, and found that the shared TFs were not significantly relevant to co-expression of divergent promoters among adjacent genes. CONCLUSION: Our findings suggested that the commonly shared cis-regulatory system does not solely contribute to the co-expression of adjacent gene pairs in yeast genome. Therefore we believe that during evolution yeasts have developed a sophisticated regulatory system that integrates both TF-based and non-TF based mechanisms(s) for concurrent regulation of neighboring genes in response to various environmental changes. PMID- 17910773 TI - Specific antibody response of mice after immunization with COS-7 cell derived avian influenza virus (H5N1) recombinant proteins. AB - To develop avian influenza H5N1 recombinant protein, the hemagglutinin (HA), neuraminidase (NA), matrix (M), and non-structural (NS1) of avian influenza H5N1 isolates from Thailand were engineered to be expressed in prokaryotic (E. coli) and mammalian cell (COS-7) system. The plasmid pBAD-His and pSec-His were used as vectors for these inserted genes. Mice immunized with purified recombinant proteins at concentration 50-250 mug intramuscularly with Alum adjuvant at week 0, week 2, and week 3 showed a good immunogenicity measured by ELISA and neutralization assay. The HA and NS recombinant proteins produced in COS-7 cells can induce specific antibody titer detected by neutralization assay significantly higher than corresponding recombinant proteins produced in E. coli system. The antibody produced in immunized mice could neutralize heterologous avian influenza virus determined by micro-neutralization assay. This study shows that avian influenza virus H5N1 recombinant proteins produced in mammalian cell system were able to induce neutralizing antibody response. PMID- 17910774 TI - Leiomyosarcoma of the adrenal vein: a novel approach to surgical resection. AB - BACKGROUND: Leiomyosarcomas typically originate within smooth muscle cells. Leiomyosarcomas arising from the adrenal vein are rare malignancies associated with delayed diagnosis and poor prognosis. The most common vascular site of origin is the inferior vena cava. CASE PRESENTATION: This is a 64-year old woman who presented with a 13 x 6.5 x 6.6 cm heterogeneous mass arising in the region of the right adrenal gland and extending into the inferior vena cava (IVC) and the right atrium. Biochemical evaluation excluded a functional tumor of the adrenal gland, and multiple tumor markers were negative. We present the novel use of deep hypothermic circulatory arrest (DHCA) in the resection of an adrenal vein leiomyosarcoma extending into the right atrium. The patient remains free of disease ten months after surgery. DHCA afforded a bloodless operative field for optimal resection of disease from within the IVC. CONCLUSION: The diagnosis of leiomyosarcomas of the adrenal vein is one of exclusion and involves preoperative radiological imaging and biochemical evaluation to exclude other functional tumors of the adrenal gland. Aggressive surgical resection is associated with improved survival and may be best achieved via collaboration among different surgical subspecialties. PMID- 17910775 TI - Program of active aging in a rural Mexican community: a qualitative approach. AB - BACKGROUND: Education is one of the key elements in the promotion of a thorough paradigm for active aging. The aim of this study is to analyze factors that contribute the empowerment of older adults in a rural Mexican community and, thus, promote active aging. METHODS: The study was conducted in a rural Mexican community (Valle del Mezquital), based on an action-research paradigm. One hundred and fifty-five elderly subjects with elementary school education participated in a formal training program promoting gerontological development and health education. Participants in turn became coordinators of mutual-help groups (gerontological nucleus) in Mexico. In-depth interviews were carried out to assess the empowerment after training for active aging. RESULTS: It was found that there was an increasing feeling of empowerment, creativity and self fulfillment among participants. Among the main factors that positively influenced training of the elderly toward active aging were the teaching of gerontology topics themselves; besides, their motivation, the self-esteem, the increased undertaking of responsibility, the feeling of belonging to the group, and the sharing of information based on personal experience and on gerontological knowledge. CONCLUSION: The main factors that contribute to empowerment of older adults in a rural Mexican community for participate in active aging programs are the training and teaching of gerontology topics themselves; besides, their interest, experience and involvement. PMID- 17910776 TI - A six-year descriptive analysis of hospitalisations for ambulatory care sensitive conditions among people born in refugee-source countries. AB - BACKGROUND: Hospitalisation for ambulatory care sensitive conditions (ACSHs) has become a recognised tool to measure access to primary care. Timely and effective outpatient care is highly relevant to refugee populations given the past exposure to torture and trauma, and poor access to adequate health care in their countries of origin and during flight. Little is known about ACSHs among resettled refugee populations. With the aim of examining the hypothesis that people from refugee backgrounds have higher ACSHs than people born in the country of hospitalisation, this study analysed a six-year state-wide hospital discharge dataset to estimate ACSH rates for residents born in refugee-source countries and compared them with the Australia-born population. METHODS: Hospital discharge data between 1 July 1998 and 30 June 2004 from the Victorian Admitted Episodes Dataset were used to assess ACSH rates among residents born in eight refugee-source countries, and compare them with the Australia-born average. Rate ratios and 95% confidence levels were used to illustrate these comparisons. Four categories of ambulatory care sensitive conditions were measured: total, acute, chronic and vaccine preventable. Country of birth was used as a proxy indicator of refugee status. RESULTS: When compared with the Australia-born population, hospitalisations for total and acute ambulatory care sensitive conditions were lower among refugee born persons over the six-year period. Chronic and vaccine-preventable ACSHs were largely similar between the two population groups. CONCLUSION: Contrary to our hypothesis, preventable hospitalisation rates among people born in refugee-source countries were no higher than Australia-born population averages. More research is needed to elucidate whether low rates of preventable hospitalisation indicate better health status, appropriate health habits, timely and effective care seeking behaviour and outpatient care, or overall low levels of health care seeking due to other more pressing needs during the initial period of resettlement. It is important to unpack dimensions of health status and health care access in refugee populations through ad-hoc surveys as the refugee population is not a homogenous group despite sharing a common experience of forced displacement and violence-related trauma. PMID- 17910777 TI - Primique: automatic design of specific PCR primers for each sequence in a family. AB - BACKGROUND: In many contexts, researchers need specific primers for all sequences in a family such that each primer set amplifies only its target sequence and none of the others, e.g. to detect which transcription factor out of a family of very similar proteins that is present in a sample, or to design diagnostic assays for the identification of pathogen strains. RESULTS: This paper presents primique, a new graphical, user-friendly, fast, web-based tool which solves the problem: It designs specific primers for each sequence in an uploaded set. Further, a secondary set of sequences not to be amplified by any primer pair may be uploaded. Primers with high sequence similarity to non-target sequences are selected against. Lastly, the suggested primers may be checked against the National Center for Biotechnology Information databases for possible mis-priming. CONCLUSION: Results are presented in interactive tables, and various primer properties are listed and displayed graphically. Any close match alignments can be displayed. Given 30 sequences, the running time of primique is about 20 seconds.primique can be reached via this web address: http://cgi www.daimi.au.dk/cgi-chili/primique/front.py. PMID- 17910778 TI - Tackling health inequalities: moving theory to action. AB - BACKGROUND: This paper reports on health inequalities awareness-raising workshops conducted with senior New Zealand health sector staff as part of the Government's goal of reducing inequalities in health, education, employment and housing. METHODS: The workshops were based on a multi-method needs assessment with senior staff in key health institutions. The workshops aimed to increase the knowledge and skills of health sector staff to act on, and advocate for, eliminating inequalities in health. They were practical, evidence-based, and action oriented and took a social approach to the causes of inequalities in health. The workshops used ethnicity as a case study and explored racism as a driver of inequalities. They focused on the role of institutionalized racism, or racism that is built into health sector institutions. Institutional theory provided a framework for participants to analyse how their institutions create and maintain inequalities and how they can act to change this. RESULTS: Participants identified a range of institutional mechanisms that promote inequalities and a range of ways to address them including: undertaking further training, using Maori (the indigenous people) models of health in policy-making, increasing Maori participation and partnership in decision making, strengthening sector relationships with iwi (tribes), funding and supporting services provided 'by Maori for Maori', ensuring a strategic approach to intersectoral work, encouraging stronger community involvement in the work of the institution, requiring all evaluations to assess impact on inequalities, and requiring the sector to report on progress in addressing health inequalities. The workshops were rated highly by participants, who indicated increased commitment to tackle inequalities as a result of the training. DISCUSSION: Government and sector leadership were critical to the success of the workshops and subsequent changes in policy and practice. The use of locally adapted equity tools, requiring participants to develop action plans, and using a case study to focus discussion were important to the success for the training. Using institutional theory was helpful in analysing how drivers of inequalities, such as racism, are built into health institutions. This New Zealand experience provides a model that may be applicable in other jurisdictions. PMID- 17910779 TI - Atsuyoshi Takao. PMID- 17910780 TI - The impact, and surgical implications, of isolated anomalous connection of one pulmonary vein. AB - Partially anomalous pulmonary venous connection of a solitary pulmonary vein in the setting of an intact atrial septum is often subclinical, and the indications for surgical repair are controversial. Here we describe a patient who developed a significant shunt over a period of 10-years. Flow through the anomalously connected pulmonary segment depends on the difference of pressure between the right and left atrium, and may increase with age. PMID- 17910781 TI - Organisational barriers to thrombolysis treatment of acute ischaemic stroke. AB - BACKGROUND: Intravenous thrombolysis with fibrinolytic drugs such as alteplase is not implemented widely in any country although the treatment is both effective and cost-effective in selected patients within a 3-h window after acute ischaemic stroke. The purpose of the present study was to describe the organisational barriers to delivery of thrombolysis for acute ischaemic stroke with special regard to the Danish healthcare system. METHOD: Systematic and unsystematic searches of medical, economic and grey literature on organisational barriers to thrombolysis treatment were performed in Cochrane, PubMed, EMBASE, Cinahl, Econlit, NHS EED, SvedMed+ and the Health Technology Assessment (HTA) database. The search periods were 1996-2006. FINDINGS: Three main types of literature on organisational barriers were found: medical literature including HTA reports on barriers related to the 3-h window, economic literature on barriers related to the lack of capacity to provide the treatment on a 24-h basis, and grey literature/policy papers on standards and demands to the hospitals and healthcare systems who implements the treatment. CONCLUSION: Information on organisational barriers can be extracted from different types of literature (medical, economic and grey literature/policy papers), but organisational barriers are most often not the primary study objective in the relevant literature. This review showed a broad spectrum of possible organisational barriers to the delivery of thrombolysis treatment of acute ischaemic stroke. PMID- 17910785 TI - Prism-based infrared spectrographs using modern-day detectors. AB - A comparison of prism-based spectrographs to grating-based spectrographs is made when each of the systems is coupled to a modern-day liquid-nitrogen-cooled photovoltaic array detector. A comparison of the systems is also made using a room-temperature microbolometer array detector. Finally, infrared microspectroscopy of samples whose size is approximately 10 micrometers will be demonstrated using a prism spectrograph outfitted with both types of detectors. The results of the study show that prism-based spectrographs offer an economical alternative to grating-based systems when spectral coverage is more critical than spectral resolution. The results also demonstrate that spectra with good signal to-noise ratios can be collected on any of the systems with a total integration time of 10 seconds or less. PMID- 17910784 TI - Coherent anti-Stokes Raman scattering microscopy. PMID- 17910787 TI - Study of thermal dynamics of defatted bovine serum albumin in D2O solution by Fourier transform infrared spectra and evolving factor analysis. AB - Fourier transform infrared (FT-IR) spectra have been measured for defatted bovine serum albumin (BSA) in D(2)O with a concentration of 2.0 wt % over a temperature range of 26-90 degrees C and the corresponding difference spectra have been calculated by subtracting the contribution of D(2)O at the same temperature. Evolving factor analysis (EFA) by selecting two factors and three factors has been employed to analyze the temperature-dependent difference IR spectra in the 1700-1600 cm(-1) spectral region of the defatted BSA in D(2)O solution. Three factor EFA has been employed to determine the distinction of the three protein species involved in the process of temperature elevation: native, transitional, and denatured protein. The temperature profiles obtained from three-factor EFA indicate that heat-induced conformational change in the secondary structures of defatted BSA in D(2)O undergoes two two-state transitions, a drastic transition and a slight transition, which occur in the temperature ranges of 68-82 degrees C and 56-76 degrees C, respectively. PMID- 17910788 TI - Two-dimensional attenuated total reflection infrared and near-infrared correlation study of the structure of butyl alcohol/water mixtures. AB - The effect of temperature on attenuated total reflection infrared (ATR-IR) and near-infrared (NIR) transmission spectra of pure butan-1-ol, butan-2-ol, 2-methyl propan-1-ol, 2-methyl-propan-2-ol, and mixtures with a small water content (X(H2O) 95% of the added psychotropics, the most common being olanzapine and haloperidol. The relative risk (quetiapine vs. risperidone) for antipsychotic polypharmacy was 1.90 (p = 0.001; 95% CI 1.29, 2.80). The mean projected cost of additional antipsychotics per randomized patient during the additive-therapy phase was $57.03 in the risperidone group and $101.64 in the quetiapine group (p < 0.01). CONCLUSIONS: The results confirm earlier reports of higher rates of polypharmacy with quetiapine than with risperidone. The findings also reveal substantial between-treatment differences in costs associated with polypharmacy. Limitations of the study include that the study was of short duration and that a high proportion of patients were recruited from countries other than the United States. PMID- 17910803 TI - Patient baseline characteristics in an open-label multinational study of betahistine in recurrent peripheral vestibular vertigo: the OSVaLD study. AB - OBJECTIVES AND METHODS: OSVaLD (Observational Study in patients suffering from recurrent peripheral vestibular Vertigo to Assess the effect of betahistine 48 mg/day on quality of Life and Dizziness symptoms) is a 3-month, open-label, multi national post-marketing surveillance study of betahistine 48 mg/day in the management of patients with vertigo of less than 5 years in duration. The aim of the study is to examine the burden of disease associated with vertigo, as determined by scores on the Dizziness Handicap Inventory (DHI), Short Form-36 (SF 36) questionnaire and the Hospital Anxiety and Depression Scale (HADS). Changes in DHI, SF-36 and HADS scores between baseline and 3 months are used to assess the therapeutic effects of betahistine. RESULTS: Participants (n = 2037) have been recruited from 13 countries in four continents (North and South America, Asia and Europe), representing a wide range of cultural and linguistic traditions. Approximately two-thirds of the patients are women. Sixty per cent of patients have diagnoses of peripheral vestibular vertigo of unknown pathology or benign paroxysmal positional vertigo; 13% have a diagnosis of Meniere's disease. All three of the instruments used characterize this as a population with extensive vertigo-attributable morbidity at baseline. The mean DHI score of the population is 63.7 +/- 15.7 (DHI scale: 0 = no handicap; 100 = major self perceived handicap), SF-36 scores in all domains are below the population average for the USA and the HADS indicated that > 50% of patients exhibit symptoms of anxiety or depression or both, including 9% who have severe manifestations of either or both conditions. CONCLUSIONS: This report describes the design and implementation of OSVaLD and presents baseline demographic and clinical features of the patients. Full results of the study, anticipated in 2007, will provide more details about the manifestations of vertigo in routine practice and the response to betahistine. PMID- 17910804 TI - Medication patterns and costs associated with olanzapine and other atypical antipsychotics in the treatment of bipolar disorder. AB - OBJECTIVE: Atypical antipsychotics are playing an increasing role in the treatment of bipolar disorder. The objective of this study was to assess the medication treatment patterns and costs associated with different atypical antipsychotics. METHODS: PharMetrics Integrated Database for medical and pharmacy claims was used to assess medication patterns and healthcare costs associated with atypical antipsychotics in the treatment of bipolar disorder. Patients who initiated on olanzapine, risperidone, quetiapine or ziprasidone as monotherapy or in combination with other bipolar medications between 01/2003 and 01/2004 were followed for 1 year. Pair-wise group comparisons were made between olanzapine and other atypical antipsychotics using Wilcoxon without adjustment, log linear regression model with adjustment, and propensity score-adjusted bootstrapping methods. RESULTS: Among 1516 patients with bipolar disorder, olanzapine (n = 507, 51%) was significantly (p < 0.01) more likely to be initiated as the mono-bipolar medication than risperidone (n = 424, 40%), quetiapine (n = 463, 36%) or ziprasidone (n = 122, 25%). Post-initiation, olanzapine was used as the mono bipolar medication for significantly (p < 0.01) more days (73.4) than risperidone (52.9), quetiapine (56.2) and ziprasidone (36.6). Annual healthcare costs incurred by patients with bipolar disorder varied from $14,216 for risperidone, $15,208 for olanzapine, $18,087 for quetiapine to $18,729 for ziprasidone treatments. No statistically significant differences in the annual healthcare costs were observed between olanzapine and risperidone treatments. Statistically significant differences between olanzapine and quetiapine were observed in two of the three models compared (p < 0.01, Wilcoxon; p = 0.024, log linear; p = 0.390, propensity score-adjusted bootstrapping) and between olanzapine and ziprasidone in one of the three models (p < 0.01, Wilcoxon; p = 0.068, log linear; p = 0.394, propensity score-adjusted bootstrapping). LIMITATIONS: Those arising from the data source and nature of retrospective assessments. Potential bias may also exist due to the presence of confounding factors and unobserved conditions and characteristics. As such, results of this study need to be considered in the context of its limitations and generalizability should be reserved to similar patient populations. CONCLUSIONS: Antipsychotic medication use patterns were statistically significantly different among atypical antipsychotics in the usual treatment of bipolar disorder. Olanzapine appears to be more likely used as a monobipolar medication compared with risperidone, quetiapine, and ziprasidone. The annual healthcare costs associated with the treatment of bipolar disorder by olanzapine and risperidone were similar, and the costs of these treatments were lower than with quetiapine or ziprasidone. PMID- 17910805 TI - Erythropoietic growth factors for children with cancer: a systematic review of the literature. AB - OBJECTIVE: To review evidence on the use of erythropoietic stimulating agents (erythropoietin or darbepoetin) in children with cancer. METHODS: A systematic review of the published literature was performed using MEDLINE (1966-July 2007) and references from a Cochrane systematic review (focusing mainly on adults) published in 2006. RESULTS: The review identified 12 studies, comprising five randomized trials, six case control studies and one open-label, dose-escalation study. All the studies that used adequate doses of recombinant human erythropoietin (rhEPO) (usually 150 IU/kg three times per week) demonstrated benefits for rhEPO except for one study in which rhEPO was added to G-CSF in children with high-risk neuroblastoma. Despite the heterogeneity of the populations studied, in terms of age, tumour type and chemotherapy regimen, rhEPO use was associated with consistent benefits in terms of reduced transfusion requirements and improved haematological parameters. Only one case of darbepoietin use was reported. CONCLUSIONS: While more studies are required, it appears that rhEPO is safe in this vulnerable patient group and can benefit children with cancer by preventing or ameliorating anaemia. PMID- 17910806 TI - Pharmaceutical applications for catanionic mixtures. AB - Mixtures of oppositely charged surfactants, so called catanionic mixtures, are a growing area of research. These mixtures have been shown to form several different types of surfactant aggregates, such as micelles of various forms and sizes, and lamellar structures, such as vesicles. In this review, a short introduction to the field of catanionic mixtures is presented and the pharmaceutical possibilities offered by such mixtures are reviewed. There are several interesting ideas on how to apply catanionic mixtures to improve the delivery of, for example, drug compounds and DNA, or for HIV treatment. PMID- 17910807 TI - Prediction of human pharmacokinetics--gut-wall metabolism. AB - Intestinal mucosal cells operate with different metabolic and transport activity, and not all of them are involved in drug absorption and metabolism. The fraction of these cells involved is dependent on the absorption characteristics of compounds and is difficult to predict (it is probably small). The cells also appear comparably impermeable. This shows a limited applicability of microsome intrinsic clearance (CL(int))-data for prediction of gut-wall metabolism, and the difficulty to predict the gut-wall CL (CL(GW)) and extraction ratio (E(GW)). The objectives of this review were to evaluate determinants and methods for prediction of first-pass and systemic E(GW) and CL(GW) in man, and if required and possible, develop new simple prediction methodology. Animal gut-wall metabolism data do not appear reliable for scaling to man. In general, the systemic CL(GW) is low compared with the hepatic CL. For a moderately extracted CYP3A4-substrate with high permeability, midazolam, the gut-wall/hepatic CL-ratio is only 1/35. This suggests (as a general rule) that systemic CL(GW) can be neglected when predicting the total CL. First-pass E(GW) could be of importance, especially for substrates of CYP3A4 and conjugating enzymes. For several reasons, including those presented above and that blood flow based models are not applicable in the absorptive direction, it seems poorly predicted with available methodology. Prediction errors are large (several-fold on average; maximum approximately 15-fold). A new simple first-pass E(GW)-prediction method that compensates for regional and local differences in absorption and metabolic activity has been developed. It has been based on human cell in-vitro CL(int) and fractional absorption from the small intestine for reference (including verapamil) and test substances, and in-vivo first-pass E(GW)-data for reference substances. First-pass E(GW)-values for CYP3A4-substrates with various degrees of gastrointestinal uptake and CL(int) and a CYP2D6-substrate were well-predicted (negligible errors). More high quality in-vitro CL(int)- and in-vivo E(GW)-data are required for further validation of the method. PMID- 17910808 TI - Development and in-vivo evaluation of insulin-loaded chitosan phthalate microspheres for oral delivery. AB - Novel chitosan phthalate microspheres containing insulin were prepared by emulsion cross-linking technique. The feasibility of these microspheres as oral insulin delivery carriers was evaluated. The pH-responsive release behaviour of insulin from microspheres was analysed. The ability of chitosan phthalate-insulin microspheres to enhance intestinal absorption and improve the relative pharmacological availability of insulin was investigated by monitoring the plasma glucose and insulin level of streptozotocin-induced diabetic rats after oral administration of microspheres at insulin dose of 20 IU kg(-1). In simulated gastric fluid (pH 2.0), insulin release from the microspheres was very slow. However, as the pH of the medium was changed to simulated intestinal fluid (pH 7.4), a rapid release of insulin occurred. The relative pharmacological efficacy for chitosan phthalate microspheres (18.66 +/- 3.84%) was almost four-fold higher than the efficacy of the chitosan phthalate-insulin solution administration (4.08 +/- 1.52%). Chitosan phthalate microspheres sustained the plasma glucose at pre diabetic level for at least 16 h. These findings suggest that the microsphere is a promising carrier as oral insulin delivery system. PMID- 17910809 TI - Formulation and in-vivo evaluation of L-cysteine chewing gums for binding carcinogenic acetaldehyde in the saliva during smoking. AB - Cigarette smoke contains toxic amounts of acetaldehyde that dissolves in saliva, posing a significant risk of developing oral, laryngeal and pharyngeal carcinomas. L-cysteine, a non-essential amino acid, can react covalently with carcinogenic acetaldehyde to form a stable, non-toxic 2-methylthiazolidine-4 carboxylic acid. The main aim of this study was to find out whether it is possible to develop a chewing gum formulation that would contain cysteine in amounts sufficient to bind all the acetaldehyde dissolved in saliva during the smoking of one cigarette. The main variables in the development process were: (1) chemical form of cysteine (L-cysteine or L-cysteine hydrochloride), (2) the amount of the active ingredient in a gum and (3) manufacturing procedure (traditional or novel compression method). Saliva samples were taken over 2.5 minutes before smoking and since smoking was started for 2.5 minutes periods for 10 minutes. During a five minutes smoking period with a placebo chewing gum, acetaldehyde levels increased from 0 to 150-185 microM. Once smoking was stopped, the acetaldehyde levels quickly fell to levels clearly below the in-vitro mutagenic level of 50 microM. All chewing gums containing cysteine could bind almost the whole of the acetaldehyde in the saliva during smoking. However, elimination of saliva acetaldehyde during smoking does not make smoking completely harmless. Cysteine as a free base would be somewhat better than cysteine hydrochloride due to its slower dissolution rate. Both traditional and direct compression methods to prepare chewing gums can be utilized and the dose of L-cysteine required is very low (5 mg). PMID- 17910810 TI - Stealth and non-stealth nanocapsules containing camptothecin: in-vitro and in vivo activity on B16-F10 melanoma. AB - Camptothecin (CPT) is an alkaloid that displays considerable antitumour activity, but clinical use has been limited by its poor water solubility and the instability of the lactone moiety (active form) in physiological media. We have therefore formulated the drug into nanocarrier systems in an attempt to improve its therapeutic properties. This study evaluates the effect of intraperitoneally administered stealth and non-stealth nanocapsules containing CPT on lung metastatic spread in mice inoculated with B16-F10 melanoma cells, and on the cytotoxic activity against B16-F10 melanoma cells in-vitro. Poly (D,L-lactide) PLA (non-stealth) and methoxy polyethylene glycol-(D,L-lactide) (PLA-PEG) (stealth) nanocapsules (49 and 66.6 kDa) were prepared by interfacial deposition of preformed polymer. CPT, as free drug or as drug-loaded nanocapsules, was administrated at a dose of 0.5 mg kg(-1) at 3-day intervals for 17 days. Free drug and CPT-loaded nanocapsules reduced the number of metastatic nodules by 45.09-91.76% (P < 0.05 vs positive control). However, only CPT-loaded PLA-PEG 49 kD nanocapsules significantly decreased the number of lung metastases when compared with free drug (P < 0.05). The administration of CPT-loaded nanocapsules and free drug did not result in neutropenia at the administered dose. The improved effectiveness of pegylated nanocapsules was attributed to protection of the drug by nanoencapsulation and to reduced uptake of particles by macrophages located in the lymph nodes. This assumption was supported by the in-vitro study, in which both PLA and 49 kDa PLA-PEG nanocapsules containing CPT were more cytotoxic than the free drug against B16-F10 melanoma cells. PMID- 17910811 TI - In-vitro controlled release of doxorubicin from silica xerogels. AB - This study aimed at the development of a novel silica xerogel matrix as a delivery tool for an anti-cancer drug. Doxorubicin was incorporated as a hydrochloride salt during hydrolysis and polycondensation of tetraethylorthosilicate (TEOS) in the sol-gel process. The effect of sol-gel synthesis parameters (drug concentration, size of the device and lyophilizing process) on the release rate of the drug were investigated. In addition, dissolution rate, as well as weight loss of silica xerogel, was evaluated. In general, both the lyophilizing process of xerogels and the increase in size of non-lyophilizing device significantly decrease both the rate of drug release and the rate of dissolution of matrix. The overall release process was found to be governed by diffusion control and simultaneous zero-order dissolution of the xerogel. PMID- 17910812 TI - Is bicarbonate buffer suitable as a dissolution medium? AB - The objectives of this study were to compare two methods for the preparation of bicarbonate buffer, and to compare media prepared with bicarbonate buffer with commonly used biorelevant and pharmacopoeial media in terms of their suitability for dissolution testing. The various media were compared with regard to ease of preparation, robustness and reproducibility of composition. The dissolution of three formulations of a typical Biopharmaceutical Classification System Class II drug (BIXX) was compared in bicarbonate buffer, standard phosphate buffer, a biorelevant buffer (fasted-state simulating intestinal fluid, FaSSIF) and a modified FaSSIF prepared with bicarbonate buffer. The bicarbonate buffer used for dissolution testing was produced by supplying carbon dioxide to a saline solution (0.9% NaCl, to which 12 or 42 mmol NaOH had been added). The bicarbonate buffer had to be prepared in-situ, which proved to be time-consuming, and the pH stability of the bicarbonate buffer could only be maintained under constant CO2 supply. To minimize the mechanical stress caused by inflow and evaporation of gas, the carbon dioxide was supplied above the medium during the dissolution test. Despite taking these measures, use of bicarbonate buffer led to less reproducible dissolution results than the phosphate buffers commonly used to prepare compendial media and FaSSIF, with coefficient of variance values 1.5- to 5-times higher in bicarbonate buffer. It was concluded that although a bicarbonate buffer system would be physiologically relevant for the fasted state in the small intestine, its suitability for dissolution testing is restricted by lack of practicability and poor reproducibility of results. PMID- 17910814 TI - Suppressive activity of fexofenadine hydrochloride on nitric oxide production in vitro and in-vivo. AB - The aim of this study was to examine the effect of fexofenadine hydrochloride (FEX), a histamine H1-receptor antagonist, on nitric oxide (NO) production in vitro and in-vivo. Nasal fibroblasts (5 x 10(5) cells per mL) were stimulated with 25 ng mL(-1) tumour necrosis factor-alpha in the presence of various concentrations of FEX. NO levels in 24-h-culture supernatants were measured by the Griess method and levels of inducible nitric oxide synthase (iNOS) mRNA levels in 12-h-cultured cells were measured by ELISA. FEX at more than 0.5 microg mL(-1) suppressed NO production from fibroblasts by inhibiting expression of iNOS mRNA. We also examined whether FEX could suppress NO production induced by lipopolysaccharide (LPS) stimulation in-vivo. BALB/c mice were treated with 5.0 mg kg(-1) LPS i.p. after daily oral doses of FEX, 1.0 mg kg(-1), for 1-3 weeks. Plasma was obtained 6 h later and NO levels measured by the Griess method. Expression of iNOS mRNA in lung tissues was measured by ELISA 6 h after LPS injection. Oral administration of FEX for 2 and 3 weeks, but not 1 week, significantly suppressed NO levels in plasma through the inhibition of iNOS mRNA expression, which were enhanced by LPS stimulation. These results suggest that the attenuating effect of FEX on NO production may be of therapeutic benefit in allergic diseases. PMID- 17910813 TI - Carvedilol increases ciclosporin bioavailability by inhibiting P-glycoprotein mediated transport. AB - Carvedilol is often used to treat hypertension and for prophylaxis in vascular sclerosis in renal transplant recipients, who require concomitant treatment with ciclosporin. However, there are few reports regarding the pharmacokinetic interactions between carvedilol and ciclosporin. We have investigated the potential effects of carvedilol on the pharmacokinetics of ciclosporin, and examined the inhibitory effects of carvedilol on P-glycoprotein-mediated transcellular transport using Caco2 cells. Ciclosporin alone or with carvedilol was orally or intravenously administered to rats. The oral administration of carvedilol (10 mg kg(-1)) with ciclosporin (10 mg kg(-1)) increased the whole blood concentration of ciclosporin. When ciclosporin (3 mg kg(-1)) was intravenously administered with carvedilol (3 mg kg(-1)), there was no difference in the whole blood ciclosporin concentration between administration with and without carvedilol. Co-administration with carvedilol increased ciclosporin bioavailability from 33% to 70%. In Caco2 cells, carvedilol caused a concentration-dependent increase in the intracellular accumulation of ciclosporin, and its effect was comparable with that of verapamil. Carvedilol considerably raised the concentration of ciclosporin in the blood and this interaction was associated with the absorption phase of ciclosporin. This interaction was caused by the inhibition of P-glycoprotein-mediated transport by carvedilol in the intestine. PMID- 17910815 TI - Theogallin and L-theanine as active ingredients in decaffeinated green tea extract: II. Characterization in the freely moving rat by means of quantitative field potential analysis. AB - The model Tele-Stereo-EEG (continuous recording of intracerebral field potentials in the freely moving rat to produce an electropharmacogram) has been used to see if L-theanine- and theogallin-enriched decaffeinated green tea extract would change electrical brain activity after oral administration, to provide proof of access of active components to the brain via the blood-brain barrier. Baseline recording (45 min) was followed by a 5-h recording session after oral ingestion of the extract or single components: L-theanine, theogallin and quinic acid, a suggested metabolite of theogallin. Power spectra from Fast Fourier Transformed (FFT) field potential changes were divided into six frequency bands (delta, theta, alpha1, alpha2, beta1 and beta2). No effects could be measured using a saline solution for control purposes. Oral administration of 75 mg kg(-1) total extract led to power decreases mainly in delta and alpha2 frequencies during the first hour. This pattern has been observed in the presence of stimulatory synthetic compounds. Oral administration of 30 mg kg(-1) L-theanine led to power decreases of nearly all frequencies, being more pronounced during the second and following hours in comparison with the first hour. Ingestion of 20 mg kg(-1) theogallin also showed a power decreasing effect on cortical activity. Its possible metabolite quinic acid (10 mg kg(-1), p.o.) also produced decreases in delta, alpha2 and beta1 frequencies. Measurement of motion resulted in an increase during the first hour in the presence of theogallin and L-theanine. A tendential decrease was observed in the presence of L-theanine during the last hour at its presumably highest plasma levels. The results with the administration of the total extract provided evidence for the maior involvement of L-theanine and theogallin (or its presumable metabolite quinic acid) in its action, since no other active compounds were present in the extract. These compounds could be classified by comparison with reference drugs using discriminant analysis as being antidepressive and cognition enhancing, respectively. The extract appeared among those drugs having stimulatory effects. PMID- 17910816 TI - Effect of orally administered Eriobotrya japonica seed extract on allergic contact dermatitis in rats. AB - The anti-allergic activity of Eriobotrya japonica seeds extract (ESE) was investigated. Oral administration of ESE dramatically inhibited ear swelling due to allergic contact dermatitis caused by repeated application of two antigens, 4 ethoxymethylene-2-phenyl-2-oxazolin-5-one (oxazolone) and dinitrofluorobenzene (DNFB), respectively. The increase of histamine content in inflamed ear tissue induced by oxazolone and DNFB was significantly antagonized by orally administered ESE. Eosinophil peroxidase and myeloperoxidase activity in both models was suppressed by orally administered ESE. Tumour necrosis factor-alpha in the inflamed region caused by repeated application of DNFB was also significantly suppressed. The findings suggest that ESE may be effective for treating allergic contact dermatitis. PMID- 17910817 TI - Gastroprotective activity of the hydroalcoholic extract obtained from Polygala paniculata L. in rats. AB - The possible gastroprotective effects of the hydroalcoholic extract of Polygala paniculata in rats have been evaluated. We have investigated the effects of this hydroalcoholic extract on acute lesions induced by ethanol (70%, p.o.) and indomethacin (20 mg kg(-1), s.c.). Its influence on mucus secretion was investigated, measured as the amount of Alcian blue dye estimated by colorimetry, and antisecretory effects were assessed in the pylorus ligature model. The treatment of rats with a crude hydroalcoholic extract of P. paniculata (HEPP; 30, 100, 300 mg kg(-1), p.o., or 3, 10 and 30 mg kg(-1), i.p.) decreased the ulcer index, and maintained the gastric mucus production in acute gastric lesions caused by ethanol 70%. In addition, the extract partially protected the mucosa against indomethacin-induced lesions. The extract did not change the volume and acidity of gastric secretion in the pylorus-ligated rat. An additional antioxidant activity of the extract and its isolated flavonoid compound rutin, in the DPPH free radical scavenging assay, was observed. In conclusion, HEPP exhibited marked gastroprotection; these effects may have involved prostaglandins and be related to cytoprotective factors, such as antioxidant activity and maintenance of mucus production. PMID- 17910818 TI - Antispasmodic activity of licochalcone A, a species-specific ingredient of Glycyrrhiza inflata roots. AB - Licochalcone A, a species-specific and characteristic retrochalcone ingredient of Glycyrrhiza inflata root, has been shown to possess multiple bioactive properties. However, its muscle relaxant activity has not been reported previously. Licochalcone A showed a concentration-dependent relaxant effect on the contraction induced by carbachol (50% effective concentration (EC50) = 5.64 +/- 1.61 microM), KCl (EC50 5.12 +/- 1.68 microM), BaCl2 (EC50 1.97 +/- 0.48 microM) and A23187 (EC50 2.63 +/- 2.05 microM). Pretreatment with licochalcone A enhanced the relaxant effect of forskolin, an adenylyl cyclase activator, on the contraction in a similar manner to 3-isobutyl-1-methylxanthine (IBMX), a phosphodiesterase (PDE) inhibitor. Furthermore, the IC50 (22.1 +/- 10.9 microM) of licochalcone A against cAMP PDE was similar to that of IBMX (26.2 +/- 7.4 microM). These results indicated that licochalcone A may have been responsible for the relaxant activity of G. inflata root and acted through the inhibition of cAMP PDE. PMID- 17910819 TI - Prediction of human pharmacokinetics--improving microsome-based predictions of hepatic metabolic clearance. AB - Physiologically based methods generally perform poorly in predicting in-vivo hepatic CL (CL(H)) from intrinsic clearance (CL(int)) in microsomes in-vitro and unbound fraction in blood (f(u,bl)). Various strategies to improve the predictability have been developed, and inclusion of an empirical scaling factor (SF) seems to give the best results. This investigation was undertaken to evaluate this methodology and to find ways to improve it further. The work was based on a diverse data set taken from Ito and Houston (2005). Another objective was to evaluate whether rationalization of CL(H) predictions can be made by replacing blood/plasma-concentration ratio (C(bl)/C(pl)) measurements with SFs. There were apparently no or weak correlations between prediction errors and lipophilicity, permeability (compounds with low permeability missing in the data set) and main metabolizing CYP450s. The use of CL(int) class (high/low) and drug class (acid/base/neutral) SFs (the CD-SF method) gives improved and reasonable predictions: 1.3-fold median error (an accurate prediction has a 1-fold error), 76% within 2-fold-error, and a median absolute rank ordering error of 2 for CL(H) (n = 29). This approach is better than the method with a single SF. Mean (P < 0.05) and median errors, fraction within certain error ranges, higher percentage with most accurate predictions, and ranking were all better, and 76% of predictions were more accurate with this new method. Results are particularly good for bases, which generally have higher CL(H) and the potential to be incorrectly selected/rejected as candidate drugs. Reasonable predictions of f(u,bl) can be made from plasma f(u) (f(u,pl)) and empirical blood cell binding SFs (B-SFs; 1 for low f(u,pl) acids; 0.62 for other substances). Mean and median f(u,bl) prediction errors are negligible. The use of the CD-SF method with predicted f(u,bl) (the BCD-SF method) also gives improved and reasonable results (1.4-fold median error; 66% within 2-fold-error; median absolute rank ordering error = 1). This new empirical approach seems sufficiently good for use during the early screening; it gives reasonable estimates of CL(H) and good ranking, which allows replacement of C(bl)/C(pl) measurements by a simple equation. PMID- 17910820 TI - D-003 does not possess oestrogenic potential in-vivo: findings of the uterotrophic assay. AB - D-003 is a mixture of long-chain fatty acids purified from sugarcane wax that inhibits both cholesterol synthesis prior to mevalonate formation, and lipid peroxidation. D-003 has been shown to prevent bone loss and bone resorption in ovariectomized rats, and significantly improves bone resorption markers in postmenopausal women with reduced bone mineral density. As hormone-replacement therapy, D-003 displays cholesterol-lowering and anti-resorptive effects. We have studied its potential oestrogenic activity in-vivo using the uterotrophic assay. Rats were randomly distributed into five groups: a sham-operated group and four groups of ovariectomized rats, one treated with vehicle, one with D-003 (50 mg kg(-1)), one with oestradiol benzoate (30 microg kg(-1)) and one with D-003 (50 mg kg(-1)) plus oestradiol benzoate (30 microg kg(-1)). Treatments were administered for 14 days. Ovariectomy decreased the values of relative uterus weight, epithelium cell height and endometrial thickness compared with sham operated rats, and these effects were all significantly reduced with oestradiol benzoate, but not with D-003. Concurrent administration of D-003 and oestradiol benzoate had statistically similar effects on all variables as oestradiol benzoate alone. In conclusions, D-003 orally given at 50 mg kg(-1), a dose that prevents bone loss and bone resorption in ovariectomized rats, did not display oestrogenic/anti-oestrogenic activity in-vivo, as assessed in the uterotrophic assay. PMID- 17910821 TI - 3-[(Aryl)(4-fluorobenzyloxy)methyl]piperidine derivatives: high-affinity ligands for the serotonin transporter. AB - The structural requirements for high-affinity binding at the serotonin transporter (SERT) have been investigated through the preparation of some 3 [(aryl)(4-fluorobenzyloxy)methyl]piperidine derivatives. The affinity of synthesised piperidinic compounds (1-4) at the SERT was evaluated by displacement of [3H]-paroxetine binding. Derived inhibition constant (Ki) values were in the same order of magnitude as that of fluoxetine, ranging between 2 and 400 nM. To better define the profiles of these compounds as potential antidepressants, binding affinity for 5-HT1A receptors and alpha2-adrenoceptors was also investigated by competition experiments using [3H]8-hydroxy-2 (dipropylamino)tetralin ([3H]8-OH-DPAT) and [3H]rauwolscine as radiolabelled ligands, respectively. Inhibition data indicate that compounds 1-4 possess a very weak affinity for these receptors. The high affinity of compound 1 for SERT indicates that it is worth investigating further. PMID- 17910822 TI - PG-liposomes: novel lipid vesicles for skin delivery of drugs. AB - A novel type of lipid vesicles, propylene glycol-embodying liposomes or PG liposomes, composed of phospholipid, propylene glycol and water, is introduced. The new lipid vesicles were developed and investigated as carriers for skin delivery of the model drug, cinchocaine base. PG-liposomes showed high entrapment efficiency and were stable for at least one month of storage at 5 +/- 1 degree C. Preliminary in-vivo skin deposition studies, carried out using albino rabbit dorsal skin, showed that PG-liposomes were superior to traditional liposomes, deformable liposomes and ethosomes, suggesting that PG-liposomes, introduced in the current work, are promising carriers for skin delivery of drugs. PMID- 17910823 TI - [Cytoreductive surgery and intraperitoneal chemotherapy in colorectal cancer]. PMID- 17910824 TI - [New treatment of peritoneal carcinomatosis from colorectal cancer. Cytoreductive surgery and hyperthermic intraperitoneal chemotherapy]. AB - Peritoneal carcinomatosis (PC) is commonly seen in colorectal cancer and is uniformly fatal. Cytoreductive surgery (CS) combined with hyperthermic intraperitoneal chemotherapy (HIIC) is a new treatment in strictly selected patients with PC. CS includes peritonectomy procedures and resection of infiltrated viscera leaving no macroscopic tumor thicker than 2.5 mm behind. Peritoneal perfusion with mitomycin C at a temperature of 40 degrees -41 degrees C is performed at the end of surgery. The postoperative morbidity and mortality rates are 20%-30% and 4%-8% respectively. Median survival is 1-2 years and the 5 year survival is 19%. PMID- 17910825 TI - [Placing gastroduodenal tubes]. PMID- 17910826 TI - [Natalizumab (Tysabri)]. AB - Natalizumab is a humanized recombinant monoclonal antibody against alpha4 intigrin. In a large phase III study on relapsing-remitting MS Natalizumab reduced the relapse rate by 68% and the appearance of permanent neurological deficits by 42%. Currently, Natalizumab is the most effective disease-modifying treatment of relapsing-remitting MS. The clinical use of the treatment will be restricted initially due to uncertainty about the risk of opportunistic infections in long term treatment. PMID- 17910828 TI - [Vein of Galen aneurysm diagnosed in the perinatal period. A retrospective assessment]. AB - INTRODUCTION: A vein of Galen aneurysm (VGA) is a rare, congenital, intracranial vascular malformation often diagnosed in the perinatal period. In recent years there have been international reportings of good results after endovascular embolization during the first year of life. MATERIALS AND METHODS: In this paper the clinical course of 10 children born in Denmark with perinatal presentation of VGA is reviewed. RESULTS: 2 out of 4 children who underwent endovascular embolization during the first year of life survived. 1 became moderately motorically and mentally retarded--the other severely. No attempt was made to close the VGA in 6 children, either because it was impossible to stabilize them, or because other kinds of severe intracerebral damage were already present. CONCLUSION: 9 out of the 10 children with a perinatal presentation of VGA are either deceased or have severe motor function and mental damage. Despite international reportings it is still recommended to carefully consider whether or not it is ethically correct to continue treatment if a neonate with VGA cannot be haemodynamically stabilized within the 1st week of life despite intensive care, or if massive intracerebral damage is already present. If neonate is stabilized, specialized units that perform endovascular embolizations for VGA during the 1st year of life must be contacted for an assessment in preparation for treatment. PMID- 17910827 TI - [Restless legs syndrome. Definition, cause and treatment]. AB - Restless legs syndrome (RLS) is a common disorder affecting more than 2% of adults. RLS is characterized by sensory-motor phenomena predominantly in the legs, but other body parts may be involved. The symptoms are most pronounced during rest and present a characteristic diurnal variation with maximum symptoms in the evening and at night. The disease is often associated with periodic limb movements. RLS is present in an idiopathic and secondary form due to medical and other neurological diseases. Iron and DOPA are involved in the pathogenesis of the disease. The current best-documented treatment is partial dopamine-agonists and gabapentin. PMID- 17910829 TI - [Fatigue in hematological patients treated with allogeneic stem cell transplantation]. AB - INTRODUCTION: Fatigue and poor physical function are well-known symptoms in cancer patients. Allogeneic stem cell transplantation is an established treatment for certain cancer diagnoses, especially leukaemia. The aim of this study was to investigate physical function and fatigue in relation to allogeneic stem cell transplantation. MATERIALS AND METHODS: 21 patients awaiting allogeneic stem cell transplantation were included in a descriptive follow-up design. Fatigue and physical function was assessed by a standardized test programme consisting of a stair test, a test of maximum oxygen uptake and questionnaires on physical activity and fatigue prior to transplantation as well as 3 and 6 months after transplantation. RESULTS: 10 men and 11 women were included. The mean age was 39 years (19-60). Prior to transplantation patients had a low maximum oxygen uptake and reported more fatigue than the Danish general population. Physical fatigue rather than mental fatigue was pronounced both before and after transplantation. No significant changes in the parameters were found from before transplant to 6 months after transplant. CONCLUSION: The results indicate that prior to transplantation patients have poor physical function and increased physical fatigue. The symptoms seem to persist 6 months after transplantation. This could indicate a need for an extensive rehabilitation programme following transplantation. PMID- 17910830 TI - [Salmeterol and fluticason and mortality in COPD patients]. AB - In this study 6,112 COPD patients were treated with salmeterol 50 microg bid, fluticason 500 microg bid, salmeterol/fluticason combination 50/500 microg bid (SFK) or placebo for 3 years. The primary effect parameter was mortality and 875 patients had died after 3 years. Hazard ratio for death for SFK compared with placebo was 0.825, 95% confidence interval 0.681-1.002, p=0.052, or an absolute risk reduction of 2.6%. Active treatment reduced exacerbations and improved quality of life and lung function. The risk of pneumonia was higher in the fluticason and SFK treated groups than in the placebo group. PMID- 17910832 TI - [Leptomeningeal carcinomatosis--an infrequent cause of hearing loss]. AB - The causes of acquired hearing loss may be many. We present a case of sudden onset of bilateral sensorineural hearing loss due to leptomeningeal carcinomatosis. The most common clinical features and typical CSF changes are reviewed, and the potential difficulties in reaching the diagnosis are discussed. Modes of therapy and prognosis are briefly mentioned. PMID- 17910833 TI - [Laparoscopic repair of giant incisional hernia after abdominal wall reconstruction]. AB - Laparoscopic repair of giant incisional hernias, traditionally treated using the open technique with abdominal wall reconstruction, represents a development in the operative method with fewer peri- and post-operative complications. The authors present a patient with a giant incisional hernia after primary right hemipelvic chondrosarcoma and pelvic resection. The patient was treated with laparoscopic repair, in which a large prolene mesh was implanted, and the patient had an uncomplicated post-operative course. PMID- 17910834 TI - [Diagnostic accuracy of 18F-FDG PET in residual or recurrent differentiated thyroid carcinoma with high thyroglobulin and negative 131-I whole-body scan]. AB - INTRODUCTION: 18F-FDG PET has demonstrated its usefulness in detecting recurrences of differentiated thyroid cancer (DTC) with high thyroglobulin (Tg) levels and negative 131I whole-body scan (WBS); however, the number of patients analyzed is low and different studies report disparate results. OBJECTIVE: To estimate the diagnostic accuracy of 18F-FDG PET in this subgroup of patients. MATERIALS AND METHODS: Fifty eight patients (64 18F-FDG PET studies) with histologically proven DTC treated with total thyroidectomy followed by at least one session of 131I therapy for the ablation of thyroid remnants were retrospectively analyzed. Results were verified by pathology, clinical follow-up, response to treatment, or by comparison with conventional diagnostic methods. Sensitivity (Se), specificity (Sp), likelihood ratios (LR), diagnostic accuracy (DA), positive and negative predictive values (PPV, NPV), and Tg levels were calculated for patients with positive and negative 18F-FDG PET results. RESULTS: Twenty seven studies were true positives, 1 false positive, 25 true negatives, and 11 false negatives. Se was 71 %, Sp 96 %, PPV 96.4 %; NPV 69.4 %, DA 81.3 %, positive LR 17.75, and negative LR 0.3. The mean Tg level was 202.34 ng/ml in patients with positive 18F-FDG PET and 40.94 ng/ml in those with negative 18F-FDG PET; the difference between the two groups (161.4 ng/ml) was significant at p <0.05. CONCLUSIONS: 18F-FDG PET is a useful for detecting residual or recurrent DTC in patients with elevated Tg and negative 131I WBS. PMID- 17910835 TI - [Usefulness of thyroid scintigraphy in the therapeutic management of amiodarone induced hyperthyroidism]. AB - Amiodarone-induced hyperthyroidism is relatively common in iodine-deficient regions. Two types have been described: type I, caused by increased synthesis and release of thyroid hormone in individuals with underlying thyroid disease, can be treated with antithyroid drugs or radioiodine; and type II, a destructive thyroiditis responsive to corticoid therapy but not to antithyroid drugs. It can be difficult to distinguish between the two types, and cases of mixed types have been reported. OBJECTIVE: to assess the usefulness of thyroid scintigraphy in amiodarone-induced hyperthyroidism. MATERIALS AND METHODS: 27 consecutive patients (13 females) with amiodarone-induced hyperthyroidism. Mean age was 65 years (range: 39-89). All patients underwent 99mTc-pertechnectate thyroid scintigraphy and were classified according to the qualitative estimation of radiotracer uptake: type I (increased / normal uptake): 9 patients, all of whom responded to antithyroid drugs or radioiodine, except one patient with sub clinical hyperthyroidism who received no therapy; type II (very low or undetectable uptake): 13 patients, 11 of whom responded to discontinuation of amiodarone or prednisone therapy (2 patients). Hyperthyroidism was resistant in 2 patients and required antithyroid drugs or potassium perchlorate; mixed type (low uptake but with underlying thyroid pathology): 5 patients, with variable evolution; all needed antithyroid drugs, one required subtotal thyroidectomy, and another radioiodine treatment. CONCLUSION: thyroid scintigraphy can establish the correct therapeutic approach in most cases of amiodarone-induced hyperthyroidism, making it essential in the clinical management of these patients. PMID- 17910836 TI - [Diagnostic accuracy of FP-CIT SPECT in patients with parkinsonism]. AB - OBJECTIVE: To determine the diagnostic accuracy of FP-CIT SPECT in entities with and without presynaptic involvement of the nigral-striatal dopaminergic pathway in a large group of patients with movement disorders, evaluating the usefulness of quantitative analysis. MATERIALS AND METHODS: A group of 183 consecutive patients clinically diagnosed as either having or not having degenerative Parkinsonism. These results were then contrasted with those of FP-CIT SPECT to determine the diagnostic accuracy of the procedure. The specific binding index was evaluated with ROC curves. RESULTS: FP-CIT SPECT was highly accurate in the diagnosis of neurodegenerative Parkinsonism (sensitivity: 95 %, specificity: 90 %). Most of the false positive results arose in patients with vascular Parkinsonism and the false negative results in patients with Parkinson disease. ROC curve analysis of semiquantitative evaluation had a sensitivity of 83 % and specificity of 82 % with an optimal cut-off of 1.44. The area under the curve was not significantly different between patients 60 years (0.899 vs 0.884) of age. CONCLUSIONS: FP-CIT SPECT has a high degree of diagnostic accuracy for striatal dopaminergic involvement. No significant changes in diagnostic accuracy were seen with respect to patient age. PMID- 17910837 TI - [Are bone scintigraphy examinations requested in oncologic patients according to established indications?]. AB - OBJECTIVE: To determine the percentage of bone scintigraphy examinations (BS) requested according to established indications and to assess the clinical impact of the scintigraphic results. MATERIAL AND METHODS: A retrospective study was performed including BS in 117 patients (70 women and 47 men) carried out in our department during the year 2001. All patients had a primary extraosseous malignancy. The correctness of the indication of each study requested was analyzed according to established criteria from the literature. BS results were classified as positive, negative, and equivocal for metastatic disease. RESULTS: 96 out of the 117 BS were performed in patients affected with the most prevalent primary malignancies: breast (57), prostate (21), and lung (18). The remaining studies were included in a miscellaneous group (gynecological [3], colorectal [4], oropharyngeal [4], and renal malignancies [4]; lymphoma [2], melanoma [2], hemangioendothelioma [1]; and cancer of the bladder [1] or pancreas [1]). Ninety nine (85 %) of the 117 BS performed met the criteria for appropriate indication. The indication was correct in 75 % of breast, 90 % of prostate (19/21), and 100 % of lung cancers. The indication was correct in 90 % of the cases in the miscellaneous group. BS were positive in 21 patients (20 of which were confirmed). BS were equivocal in 24 patients (in 5 of whom bone metastases were confirmed). BS were negative in 72 patients (one of whom had bone metastases). The BS findings changed staging in 9 % (9/99) of the correctly indicated cases. CONCLUSION: Most BS (85 %) were indicated according to the established criteria and the clinical impact was greater in this group. PMID- 17910838 TI - Solitary focus in the liver in a thyroid cancer patient after a whole body scan with 131 iodine. AB - A 51-year-old woman diagnosed with follicular variant of papillary thyroid carcinoma underwent a total thyroidectomy followed four weeks later by an ablative dose of 3.7 GBq of 131I. A whole body scan 5 days after ablation showed an intense uptake within the thyroid bed and a focal uptake located in the right lung base or liver dome. Computed tomography examination revealed a hypodense hepatic node in segment VII resembling a liver metastasis. Histological examination after ultrasound-guided fine-needle aspiration characterized the lesion as a liver abscess. The abscess regressed after antibiotic therapy. Liver metastases from papillary thyroid carcinoma are uncommon. On the other hand, false positive findings of 131I whole body scans have been described. A focal hepatic uptake might represent a metastasis (rare in papillary carcinomas) or be related to other causes (cysts, inflammation or infection, non-thyroidal neoplasms, etc.). PMID- 17910839 TI - [Bone metastases from unknown primary. FDG-PET detection of two synchronous tumors]. AB - A 60-year-old woman presented with noncardiac chest pain over months and negative laboratory findings. Conventional imaging methods and bone scintigraphy detected bone lesions suggesting metastatic disease from an unknown primary tumor. An 18FDG-PET scan performed to orient the search for the primary tumor found focal lesions suggesting lymphoma and identified a focal thyroid lesion and a cervical lymph node accessible for biopsy. The biopsy of this lymph node incidentally detected a papillary differentiated thyroid cancer (DTC), since the existence of a non-Hodgkin's lymphoma was confirmed after a new biopsy. After confirming the presence of a lymphoma, 18FDG-PET enabled the initial staging of the tumor, the evaluation of the response to treatment, and follow-up for detection of recurrence. On the other hand, 18FDG-PET incidentally detected a DTC. PMID- 17910841 TI - Positron emission tomography/computed tomography (PET/CT) with 2-deoxy-2-[18F] fluoro-D-glucose (18FDG) in a patient with cutaneous T-cell lymphoma. PMID- 17910840 TI - [The value of PET/CT in the differential diagnosis of presacral mass]. AB - We present the case of one patient: a male with rectal cancer, who underwent computed tomography (CT) and positron emisission tomography/computed tomography (PET/CT) with 18F-FDG to comfirm local recurrence of rectal cancer. The results of PET/CT confirms that there one a recurrence of rectal cancer. PMID- 17910842 TI - [FDG-PET and influenza vaccine]. PMID- 17910843 TI - [Brown bone tumors as a manifestation of primary hyperparathyroidism]. PMID- 17910844 TI - [Parathyroid scintigraphy and radioguided surgery in primary hyperparathyroidism]. PMID- 17910845 TI - [Clinical, echocardiographic and prognostic evaluation of atrial fibrillation in patients with heart failure]. AB - BACKGROUND AND OBJECTIVE: We aimed to assess the prevalence of atrial fibrillation (AF) in a general heart failure (HF) population admitted to a HF unit, analyze the parameters associated with AF, and evaluate its prognostic significance. PATIENTS AND METHOD: 389 patients, 64 with AF at the first visit. Mean (SD) age was 65.38 (10.77) years and 72.5% were men. The main etiology was ischemic heart disease (59.9%). Mean ejection fraction (EF) was 32.25% (13%). Vital status at 2 years was available in 377 patients (97%), 314 in sinus rhythm (SR) and 63 in AF. RESULTS: The prevalence of AF was 15.8%. AF was associated with: older age, female gender, valvular and hypertensive etiology, longer time since the onset of HF symptoms, higher EF, higher left atrium diameter, degree of mitral regurgitation, and lower quality of life, but not with the NYHA functional class. The 2-years mortality (16.7%) was significantly higher in patients with AF (33.3% vs 18.4%; OR = 2.20; 95% confidence interval, 1.21-4). However, when adjusted for other relevant variables such as age, NYHA functional class, ejection fraction, sex and etiology, AF did not remain as an independent prognostic factor. The strongest mortality differences between patients with AF and those with SR where observed in ischemic heart disease and dilated cardiomyopathy. CONCLUSIONS: AF was associated mainly with age, valvular and hypertensive etiology, higher left atrium diameter and lower end-systolic left ventricular diameter. Two years mortality was significantly higher in patients with AF, although other parameters such as age and NYHA functional class had a higher prognostic value. PMID- 17910847 TI - [Detection and quantification of circulating tumor cells in peripheral blood in patients with colon cancer]. AB - BACKGROUND AND OBJECTIVE: The aim of this study is the detection and quantification of circulating tumor cells (CTC) in patients diagnosed with colon cancer and to establish whether they are related to the main clinicopathologic variables for this type of carcinoma. PATIENTS AND METHOD: Twenty-five colon cancer patients and 30 healthy volunteers were analysed. The quantification was performed using the CellSpotter Analyzer (Veridex LLC), that allows immunomagnetic isolation and immunospecific labelling of the cells for their enumeration. RESULTS: 72% of the colon cancer patients showed CTC and the mean number of cells found was 5 CTC/7.5 ml of peripheral blood. 52% of the samples contained 2 or more cells. Considering 2 cells as the cut-off point, a significant relationship with lactate dehydrogenase was found. CONCLUSIONS: This new technology which allows isolation and quantification of CTC in peripheral blood has proven to be valid for the detection of epithelial cells in colon cancer patients in every tumor stage. The results shown in this work confirm that cytokeratin 8, 18 and 19 are detected in CTC in this tumor type and will allow us to develop a protocol for the study of the relationship of quantification of theses cells and the clinical parameters involved in colon cancer. PMID- 17910846 TI - [Daily living activity in chronic obstructive pulmonary disease: validation of the Spanish version and comparative analysis of 2 questionnaires]. AB - BACKGROUND AND OBJECTIVE: Exercise limitation is a common finding in chronic obstructive pulmonary disease (COPD) patients and has an important role in disease prognosis and the use of health care services. There are no questionnaires in Spanish language that evaluate the impact of the disease in the activities of daily living in these patients. Our objective was to adapt and validate the Spanish versions of 2 standard questionnaires for assessment of physical activity: the London Chest Activity of Daily Living scale (LCADL) and the Modified Baecke Physical Activity Questionnaire (modified Baecke). PATIENTS AND METHOD: After carrying out the translation and back-translation, the 2 questionnaires were administered to 55 patients with COPD (mean age [standard deviation]: 66 [8] years; forced expiratory volume in 1 s: 1.2 [0.5] l, 38 [15]%) in 2 occasions and a conventional exercise test was performed. RESULTS: Cronbach's alpha coefficient was very high (0.98 and 0.97 for LCADL and modified Baecke, respectively; p < 0.0001 each). Intraclass correlation coefficients of the test re-test reliability was also very high (0.97 and 0.96, respectively). Mean values of the questionnaires in the 2 administrations were 20 (11) vs 19 (11) for LCADL (p = 0.26), and 15 (9) vs 14 (9) for modified Baecke (p = 0.51). The 2 questionnaires showed a significant correlation with quality of life (St. George's Respiratory Questionnaire), dyspnea score (Medical Research Council scale) and 6 min walk test. CONCLUSIONS: The Spanish versions of these 2 questionnaires to assess physical activity are relibale and they display the expected association with health related quality of life, dyspnea score, and the 6 min walk test, confirming their validity. Therefore, we propose its use to complement the assessment of physical activity in those patients with COPD in whom direct measurements of this variable are not available. PMID- 17910848 TI - [Heart failure and atrial fibrillation: dangerous friendship?]. PMID- 17910849 TI - [Daily living activity in COPD: a new parameter to be evaluated?]. PMID- 17910850 TI - [Lung transplanted patient climbing a mountain higher than 4,000 metres. Comparison with healthy climbers]. AB - BACKGROUND AND OBJECTIVE: To ascertain if a bilaterally lung transplanted patient can climb a mountain higher than 4,000 metres and to compare the evolution of his physiological parameters during the ascent with those of healthy mountaineers. SUBJECTS AND METHOD: Heart rate, blood pressure, arterial oxygen saturation (SaO2), forced vital capacity, forced expiratory volume in one second, Tiffenau test, 25-75 mesoexpiratory flow, peak flow, severity of dyspnoea (analogic score) and symptoms and signs of acute mountain sickness (lake Louise and Serrano Alcocer scores) were measured in a bilaterally lung transplanted patient and in 4 healthy mountaineers at sea level and at different altitudes during the ascent of Breithorn (4,164 m) from Zermatt in 3 days. RESULTS: All subjects attained the summit. The transplanted patient suffered from an acute mountain sickness at 2,700 m but recovered spontaneously. No other substantial differences were found during the climb between the patient's physiological parameters and those of the healthy controls. On the summit (third day) the patient's SaO2 (90%) was higher than the figure which should be theoretically expected for this altitude among non-acclimatised subjects (81%). CONCLUSIONS: Our study confirms that it is possible for a bilaterally lung transplanted patient to climb by his own effort a mountain higher than 4,000 m with no physiological changes other than those experienced by healthy mountaineers. Considering the spontaneous recovery from the acute mountain sickness and the high SaO2 on the summit of Breithorn, we conclude that lung transplantation does not necessarily prevent altitude acclimatisation. PMID- 17910851 TI - [Conciliation in medication]. PMID- 17910852 TI - [Streptococcus bovis: an emerging pathogen]. PMID- 17910854 TI - [Persistent fever due to ingestion of Hypericum perforatum (St. Johns wort)]. PMID- 17910853 TI - [Evolution of prevalence of type 2 diabetes in adult Spanish population]. AB - Type 2 diabetes has been catalogued as the epidemic of the 21st Century, both because of its increasing magnitude and its impact on cardiovascular disease, the leading cause of death in developed countries. This review analyses the evolution of the prevalence of diabetes in Spain over recent years, based on various cross sectional studies. This study shows that previous estimates have been surpassed, with 10-15% of Spanish adults estimated to have diabetes. Factors influencing this phenomenon include the change in diagnostic criteria, population aging, a lower mortality among persons with diabetes or a true increase in the incidence. PMID- 17910855 TI - [Multiple sclerosis and herpes encephalitis]. PMID- 17910856 TI - [Reversible posterior leukoencephalopathy as onset manifestation of systemic lupus erythematosus]. PMID- 17910857 TI - [Pseudopheochromocytoma as cause of severe and paroxysmal hypertension]. PMID- 17910858 TI - [Pegylated interferon alfa-induced psoriasis]. PMID- 17910859 TI - [Duodenal metastasis of a melanoma]. PMID- 17910860 TI - [Spanish radiologists and outstanding debts]. PMID- 17910861 TI - [Receptors and markers: toward a science of imaging through hybridization]. AB - Genomic- and proteomic-based imaging will enable the selection of populations and individuals "at risk"of suffering a certain disease before the clinical symptoms appear, identifying the existence, location, and extension of the bases of the disease, stratifying the risk, and making it possible to carry out and control treatments designed for each patient. In order to be efficacious, these molecular imaging techniques must generate both functional and structural information. The use of imaging probes together with advanced equipment allows numerous molecular complexes and cellular structures to be seen. Molecular imaging makes it possible to acquire, whether directly or indirectly, information about the spatial and temporal distribution of molecular or cellular processes that have not only clinical (diagnostic and therapeutic) applications but also basic (biochemical and physiological) applications. In the clinical area, nuclear medicine and radiology must work together to lead the development, implementation, and technological evaluation of molecular imaging. From this perspective, molecular hybridization techniques must progress toward the coordination of efforts, in training and in application as well as in research, to enable us to develop fully as imaging professionals in the service of healthcare. Molecular imaging provides imaging physicians with the possibility to advance substantially in early diagnosis, in the stratification of risk and prognosis, as well as in treatment monitoring in numerous biological and cellular processes related to the disease. PMID- 17910862 TI - [Quality research and the difficulties involved therein]. AB - After an evaluation of the current situation of research in our country and establishing the concept and requisites of what is considered "quality research", this article will analyze the difficulties in carrying it out (lack of appreciation of research activity and consequent lack of necessary support and insufficient funding, lack of training for researchers, lack of continuity, among others), while trying to provide possible solutions to these problems. The important role of university hospitals will play as the backbone of translational research, coordinating research and medical practice, is also considered, together with the need to reorganize these institutions for this transcendent mission. The driving forces behind research activity in our discipline and the characteristics of radiological research are analyzed, postulating the need to return to the basics (simple, practical patient-centered research, as opposed to research based on technology) and pointing out the interest in the integration of recently constituted European organisms to promote multicenter research in medical imaging (EIBIR: European Institute for Biomedical Imaging Research). PMID- 17910864 TI - [Magnetic resonance imaging of the mediastinal vessels in pediatric patients]. AB - The appearance of new and better magnetic resonance imaging (MRI) techniques have made the MRI a very important imaging method for the evaluation of thoracic vessels in pediatrics. The 3D angio-MRI using GD is capable of clearly demonstrating the morphology of the aorta and pulmonary vessels. The MRI may significantly reduce the number of angiographies needed and, in some patients, may even provide additional information to the angiography. PMID- 17910863 TI - [Case imaging: 1.--X-ray findings in the calcic bile syndrome]. PMID- 17910865 TI - [Case imaging: 2.--Scimitar syndrome]. PMID- 17910866 TI - [Vascular endoprostheses in the treatment of intracranial aneurysms: a systematic review]. AB - OBJECTIVE: To evaluate the effectiveness and safety of endovascular prostheses (stents) in the treatment of intracranial aneurysms. MATERIAL AND METHODS: We performed a systematic review of the literature using the databases Medline, Embase, HTA, and the Cochrane Library. Relevant articles were selected by reading the abstracts found in the search and applying a series of inclusion and exclusion criteria. Afterwards, the entire text was read critically and the results were extracted and analyzed. RESULTS: Only ten series fulfilled the inclusion criteria established, and different methodological limitations were observed. A total of 287 lesions were treated; 76 of these were wide-neck aneurysms, and total or nearly total occlusion was achieved in approximately 77% of cases. Among the 281 patients included, 14 deaths (4.9%) and 48 complications (17.1%) occurred; 55% of the complications were of thromboembolic origin. Moreover, in 71 cases (24.7%) there were difficulties in placing the stent, stent displacement, or other technical problems. CONCLUSIONS: Despite the data reported in this review, the lack of methodologically sound studies with long-term follow up as well as the methodological limitations and heterogeneity found in the observational studies included in this review make it impossible to reach definitive, categorical conclusions regarding the efficacy and safety of this technique. Therefore, it would be recommendable to initiate randomized clinical trials to compare this technique with other treatment options, using appropriate and homogeneous criteria for the selection of both patients and centers together with strict follow-up, registering, and evaluation of the results obtained. PMID- 17910867 TI - [US-guided localization of non-palpable breast cancer and sentinel node using 99mTechnetium-albumin colloid]. AB - OBJECTIVE: Surgery on non-palpable breast lesions is becoming increasingly common and new techniques for preoperative lesion localization have appeared. Radio guided occult lesion localization (ROLL) enables malignant or probably malignant non-palpable breast lesions to be located and biopsy of the sentinel node to be performed (SNOLL: sentinel node and occult lesion localization). MATERIAL AND METHODS: Included were 118 patients with malignant or probably malignant non palpable breast lesions visible on ultrasonography in whom radio-guided lesion resection and sentinel node biopsy were indicated. 99mTechnetium-albumin colloid was injected into the periphery of the lesion under ultrasonographic guidance and all patients underwent preoperative scintigraphy. RESULTS: From November 2001 to December 2004, 118 patients were included. All patients underwent conservative surgery, with the non-palpable lesion being located in all cases (100% lesion detection rate). The histological diagnoses were: 81 invasive ductal carcinomas (68.64%), 7 infiltrating lobular carcinomas (5.93%), 5 mixed-type carcinomas (4.24%), 17 carcinomas in situ (14.40%), and 8 other invasive carcinomas (6.78%). The sentinel node was detected in 98.41%. DISCUSSION AND CONCLUSIONS: Radio guided ROLL surgery on non-palpable lesions located under ultrasonographic guidance is a simple, fast technique that enables the lesion to be safely excised. Both ROLL and SNOLL can be carried out in the same intervention with a single ultrasound-guided injection of 99mTechnetium-albumin colloid with satisfactory results. PMID- 17910868 TI - [Calcified cerebral metastasis]. AB - A hyperdense intraparenchymal lesion on a cerebral computed tomography (CT) usually corresponds to an acute hematoma; however, it is sometimes necessary to rule out a metastatic cause. Focal calcifications in the brain are common and are most often due to granulomas (tuberculosis, cysticercosis...), hamartomas, and primary brain tumors. Cerebral metastases are the most common intracranial neoplasm; however, their rate of calcification in classic series is only approximately 1%. We report the case of a completely calcified cerebral metastasis studied by CT and magnetic resonance imaging (MRI) that was interpreted as acute hemorrhage on the first CT examination. PMID- 17910869 TI - [Case imaging: 3.--Septo-optic dysplasia]. PMID- 17910871 TI - [Vesicosigmoid fistulas secondary to diverticulitis: helical CT diagnosis]. AB - Colovesical fistulas are the most common type of vesicointestinal fistula and the most prevalent of these are vesicosigmoid fistulas. In our environment, diverticulitis is the most common cause, accounting for approximately one half of all cases. The literature describes different methods for the diagnosis of colovesical fistula, with computed tomography and cystography being the most useful. We describe two cases of vesicosigmoid fistula secondary to diverticulitis diagnosed at our center using helical computed tomography and review the clinical and radiological findings for this entity. PMID- 17910870 TI - [Localized fibrous tumor of the pleura: radiological findings]. AB - Localized fibrous tumors of the pleura are rare tumors that represent less than 5% of the pleural tumors, although they have also been described in extrathoracic locations such as the abdomen, head and neck or central nervous system. A total of 80% begin in the visceral pleura and are not related with environmental risk factors. They generally occur in patients over 50 with a mild predominance in the woman. Up to 50% of the patients are asymptomatic, so that it appears as an incidental finding on the chest X-ray. These are slow growing tumors. They are seen on the X-ray as well-defined rounded lesions dependent on the pleura and 50% are pediculated. It is important to know its radiological characteristics for its diagnosis of suspicion and correct treatment since surgical resection is generally curative. Local recurrence after the surgery is very common if it is not completely excised. PMID- 17910872 TI - [Case imaging: 4.--Ascaris lumbricoides in the bile duct: findings on the ultrasonograph and magnetic resonance imaging]. PMID- 17910873 TI - [Vein of Galen malformation in an adult: MRI diagnosis]. AB - Malformation of the vein of Galen is an uncommon intracranial vascular anomaly that is mainly seen in pediatric patients. It presents with diverse manifestations and variable severity. It consists of dilated deep venous structures in the system of Galen, fed by abnormal arteriovenous communications in the midline. Magnetic resonance imaging and magnetic resonance angiography are better than computed tomography for the study of this condition. The case reported here joins the set of rare cases in which a vein of Galen malformation is discovered in an adult. PMID- 17910874 TI - [Persistent primitive trigeminal artery associated to aneurysm of the posterior communicating artery: a case report]. AB - We report the case of a patient that presented with an intense headache caused by a subarachnoid hemorrhage. Magnetic resonance angiography demonstrated the existence of an aneurysm of the left posterior communicating artery, as well as of a persistent primitive trigeminal artery. PMID- 17910876 TI - [Primary mesenchymal chondrosarcoma of the mediastinum]. AB - Cartilaginous lesions of the mediastinum are quite rare and primary cartilage forming tumors arising within the mediastinum are even more exceptional. Radiologic findings are unspecific. However, certain characteristics can orient the diagnosis. They are presented one mediastinal chondrosarcoma case in adult patient of patho-anatomical diagnosis. The findings described entail chest radiography, computed tomography and magnetic resonance. PMID- 17910875 TI - [Cystic lymphangioma of the spleen in a patient with Klippel-Trenaunay-Weber syndrome: MRI findings]. AB - We present the case of a 22-year-old woman with Klippel-Trenaunay-Weber syndrome who presented with clinical signs and symptoms of gastrointestinal obstruction. Ultrasonography and magnetic resonance imaging showed massive splenomegaly with multiple cysts replacing the normal parenchyma of the spleen. Histologic study after splenectomy confirmed the diagnosis of cystic lymphangioma of the spleen, which is an exceptional manifestation of Klippel-Trenaunay-Weber syndrome. PMID- 17910877 TI - [Holoprosencephaly. Role of magnetic resonance imaging in prenatal diagnosis]. AB - We presented a case affected of trisomia 13 and fetal malformations. Prenatal sonography and magnetic resonance imaging were performed. The fetus suffered semilobar holoprosencephaly, craniofacial malformations and tetralogy of Fallot. A review of sonographics and magnetic resonance findings is presented emphasizing the role of magnetic resonance as a complementary imaging technique when sonography has diagnostic limitations. PMID- 17910878 TI - [Merkel cell carcinoma: a report of two cases]. AB - We present two cases of Merkel cell carcinoma (also called cutaneous neuroendocrine carcinoma) and review the relevant literature. This is a rare skin tumor that is very aggressive, with a great tendency to local recurrence and rapid metastatic spread to regional lymph nodes as well as hematogeneous dissemination. We discuss the usefulness of different imaging modalities in determining the extension of disease and in patient follow-up. PMID- 17910883 TI - Assessment of the centrosome amplification by quantification of gamma-tubulin in Western blotting. PMID- 17910884 TI - A large-scale validation of dosage analysis by robust dosage-polymerase chain reaction. AB - Epidemiological and clinical diagnostic assays benefit from accurate detection of deletions and duplications commonly missed by the conventional strategy of polymerase chain reaction (PCR) amplification and sequencing of individual exons. Robust dosage-PCR (RD-PCR) is a quantitative duplex PCR method that coamplifies a target template and an endogenous internal control (an autosomal and an X chromosomal segment) for detection of these mutations. In this study, 110 consecutive RD-PCR assays were developed and validated. The average linear regression coefficient between template copy number and product yield and the average coefficient of determination for linear correlation, R(2), were very high: 0.95 and 0.98, respectively. The accuracy of RD-PCR revealed somatic mosaicism for a deletion in the factor 9 gene. Advantages of RD-PCR include (1) high accuracy and consistency, (2) easy calibration of linearity using male and female samples, (3) use of an endogenous internal dosage control to eliminate preparation and manipulation errors, and (4) detection of gene dosage over a wide dynamic range. Deletions and duplications can be easily detected (a 2x decrease or a 1.5x increase in gene dosage). Thus, RD-PCR is a general and accurate method for detecting changes in gene dosage. PMID- 17910885 TI - Mechanisms of human gamma-globin transcriptional induction by apicidin involves p38 signaling to chromatin. AB - Histone deacetylase (HDAC) inhibitors are one of promising drugs to induce fetal hemoglobin (HbF) for treatment of sickle cell disease (SCD) and beta-thalassemia. The HDAC inhibitor apicidin was recently reported as a powerful inducer of HbF via a mechanism involving p38 signaling. In this study, we further investigated the signaling effects on the transcriptional activation of gamma-globin gene. First, we compared histone 3 (H3) acetylation patterns of approximately 70kb beta globin loci in K562 erythroid versus HeLa cells upon apicidin treatment by chromatin immunoprecipitation assays. The results showed that the level of H3 acetylation was globally increased from the LCR to the promoter of gamma-globin gene in K562 cells, but not in non-erythroid, HeLa cells. Inhibition of p38 signaling blocks the effects of apicidin-induced gamma-globin expression and H3 acetylation. In parallel, we assessed the recruitment of transcriptional complex to beta-globin locus following apicidin treatment. The binding of GATA-1, Sp1 and RNA polymerase II (pol II) were observed to increase over several regulatory regions of beta-globin locus. Inhibitor study revealed that p38 pathway was not involved in their recruitments by apicidin. Collectively, our results provide a molecular basis to elucidate the underlying mechanisms involving p38 signaling pathway in the inducement of gamma-globin transcriptional expression by apicidin. PMID- 17910886 TI - Critical intermediate steps in Clostridium sordellii lethal toxin-induced apoptosis. AB - Clostridium sordellii lethal toxin (TcsL) inactivates small GTPases via glucosylation and induces apoptosis in mammalian cells; however, signaling events that link substrate modification with modulation of the mitochondria in these cells has not been determined. Experiments in the current study examined TcsL modulation of the Akt signaling pathway and related downstream targets. Early in TcsL intoxication, cells demonstrated a dramatic decrease in phosphorylated Akt, and this event required toxin enzymatic activity. The decrease in phosphorylated Akt was followed by caspase-dependent processing of Bcl-x(L) and Bid, revealing the connection between GTPase inactivation and mitochondrial-mediated apoptosis observed in TcsL-intoxicated cells. Levels of glycogen synthase kinase-3beta declined during later times of TcsL intoxication, suggesting a second intermediate step in apoptosis. Collectively, these data provide insight into the cascade of signaling events that lead to apoptotic death of TcsL-intoxicated cells. PMID- 17910887 TI - Practical preparation of 2-azido-2-deoxy-beta-D-mannopyranosyl carbonates and their application in the synthesis of oligosaccharides. AB - 1-O-Allyloxycarbonyl (or ethyloxycarbonyl)-2-azido-2-deoxy-3-O-benzyl (or allyl, or benzoyl)-4,6-O-isopropylidene-beta-d-mannopyranose derivatives were prepared from the corresponding 2-hydroxy-beta-d-glucopyranosyl carbonates in high yields via triflation of the 2-hydroxyl group and subsequent SN2 displacement with azide ion. An N-acetyl-mannosamine-containing trisaccharide, a fragment of the putative O10 antigen from Acinetobacter baumannii, was efficiently synthesized using these derivatives. PMID- 17910888 TI - Changes in immune effort of male field crickets infested with mobile parasitoid larvae. AB - Insect immune defenses include encapsulation and the production of lysozymes and phenoloxidase. However, the highly mobile larvae of parasitoid Ormiine flies (Ormia ochracea) can evade initial encapsulation, and instead co-opt host immune responses to form a critical respiratory funnel connecting them to outside oxygen. Here we ask how field crickets (Teleogryllus oceanicus) respond immunologically to O. ochracea infestation. Host encapsulation and phenoloxidase play important roles in the formation of the respiratory funnel, so we hypothesized that decreases in these immune parameters during infestation may interfere with respiratory funnel formation and increase the likelihood of larval death. Encapsulation ability decreased after infestation with O. ochracea larvae, but phenoloxidase activity increased in both infested crickets and controls, whereas lysozyme activity decreased in infested crickets but remained constant in controls. Hosts with fewer established larvae showed greater decreases in encapsulation, and phenoloxidase activity was positively associated with the degree of larval respiratory funnel melanization. Differences between phenoloxidase and lysozyme activity in infested crickets are consistent with a trade-off within the immune system of hosts, and our results demonstrate the effects of a prior immune challenge on the ability to mount a subsequent response. PMID- 17910889 TI - Modelling of the ventricular conduction system. AB - The His-Purkinje conduction system initiates the normal excitation of the ventricles and is a major component of the specialized conduction system of the heart. Abnormalities and propagation blocks in the Purkinje system result in abnormal excitation of the heart. Experimental findings suggest that the Purkinje network plays an important role in ventricular tachycardia and fibrillation, which is the major cause of sudden cardiac death. Nowadays an important area in the study of cardiac arrhythmias is anatomically accurate modelling. The majority of current anatomical models have not included a description of the Purkinje network. As a consequence, these models cannot be used to study the important role of the Purkinje system in arrhythmia initiation and maintenance. In this article we provide an overview of previous work on modelling of the Purkinje system and report on the development of a His-Purkinje system for our human ventricular model. We use the model to simulate the normal activation pattern as well as abnormal activation patterns resulting from bundle branch block and bundle branch reentry. PMID- 17910890 TI - What are the best outcome measurements for atopic eczema? A systematic review. AB - BACKGROUND: Valid and reliable outcome measurements are a prerequisite for evidence-based practice. The comparative validity and reliability of outcome measurements for assessing atopic eczema (AE) severity is unclear. OBJECTIVE: We sought to assess the validity, reliability, sensitivity to change, and ease of use of outcome measurements for AE. We also sought to give recommendations on which outcomes to use in clinical research and for clinical monitoring. METHODS: We performed a systematic review and survey of clinical experts and patients. RESULTS: Twenty published outcome measurements were identified. There is evidence of adequate construct validity for 3 measurements (Severity Scoring of Atopic Dermatitis index [SCORAD], Eczema Area and Severity Index [EASI], and Three Item Severity Score), adequate internal consistency of 1 scale (Patient-oriented Eczema Measure [POEM]), adequate interobserver reliability of 5 measurements (Basic Clinical Scoring System; Nottingham Eczema Severity Score; Objective Severity Assessment of Atopic Dermatitis; Six Area, Six Sign Atopic Dermatitis severity score; and SCORAD), adequate test-retest reliability of 1 scale (POEM), and adequate sensitivity to change of 3 measurements (EASI, SCORAD, and Investigators' Global Atopic Dermatitis Assessment). Most outcome measurements have adequate content validity, as assessed by patients and experts. Data on the time to perform the assessment was identified for 8 outcome measurements. Only SCORAD, EASI, and POEM have been tested sufficiently and performed adequately. CONCLUSION: There are too many published outcome measures for AE. Most have not been tested properly or perform adequately when tested, and their continued use hampers scientific communication. CLINICAL IMPLICATIONS: Only SCORAD, EASI, and POEM currently perform adequately. These scales should be used in future studies. PMID- 17910892 TI - Topical tacrolimus in the treatment of atopic dermatitis--does it benefit the airways? A 4-year open follow-up. PMID- 17910895 TI - Opioids and migration, chemotaxis, invasion, and adhesion of human cancer cells. AB - This study was designed to examine the role of opioids on cell migration, chemotaxis, invasion, and adhesion, with an emphasis on whether the opioid growth factor (OGF, [Met(5)]-enkephalin) or the opioid antagonist naltrexone (NTX) impacts any or all of these processes. Drug concentrations of OGF and NTX known to depress or stimulate, respectively, cell proliferation and growth were analyzed. Three different human cancers (pancreatic, colon, and squamous cell carcinoma of the head and neck), represented by seven different cancer cell lines (PANC-1, MIA PaCa-2, BxPC-3, CAL-27, SCC-1, HCT-116, and HT-29), were evaluated. In addition, the influence of a variety of other natural and synthetic opioids on cell motility, invasion, and adhesion was assessed. Positive and negative controls were included for comparison. OGF and NTX at concentrations of 10(-4) to 10(-6)M, and dynorphin A1-8, beta-endorphin, endomorphin-1, endomorphin-2, leucine enkephalin, [D-Pen(2,5)]-enkephalin (DPDPE), [D-Ala(2), MePhe(4), Glycol(5)]-enkephalin (DAMGO), morphine, and U69,593 at concentrations of 10( 6)M, did not alter cell migration, chemotaxis, or invasion of any cancer cell line. OGF and NTX at a concentration of 10(-6)M, and incubation for 24 or 72h, did not change adhesion of these cancer cells to collagen I, collagen IV, fibronectin, laminin, or vitronectin. Moreover, all other opioids tested at 10( 6)M concentrations and for 24h had no effect on adhesion. These results indicate that the inhibitory or stimulatory actions of OGF and NTX, respectively, on cell replication and growth are independent of cell migration, chemotaxis, invasion, and adhesive properties. Moreover, a variety of other exogenous and endogenous opioids, many specific for the micro, delta, or kappa opioid receptors, also did not alter these biological processes, consonant with previous observations of a lack of effects of these compounds and their receptors on the biology of cancer cells. PMID- 17910894 TI - Vitamins A and D are potent inhibitors of cutaneous lymphocyte-associated antigen expression. AB - BACKGROUND: Cutaneous lymphocyte-associated antigen (CLA) is a surface glycoprotein expressed by skin-homing T cells. This carbohydrate moiety expressed on mucin-like surface glycoproteins, including P-selectin glycoprotein ligand 1 and CD43, confers binding activity to dermal endothelial E-selectin and is critical for T-cell recruitment to the skin. Vitamin A (retinoic acid [RA]) and the active form of vitamin D3 (1,25 dihydroxyvitamin D3 [1,25D(3)]) have been used to treat certain T cell-mediated inflammatory skin diseases, as well as cutaneous T-cell lymphomas; however, their effect on CLA expression has not been studied. OBJECTIVE: We analyzed the effects of RA and 1,25D(3) on expression of CLA and other lymphocyte-homing receptors on human T cells. METHODS: We cultured human T cells with 1,25D(3) and RA and analyzed the expression of CLA and other homing receptors. We also pretreated mice with either vitamin and then induced an antigen-dependent contact hypersensitivity response. RESULTS: Both RA and 1,25D(3) downregulated expression of the CLA and, in parallel, functional E selectin ligand. Whereas RA increased expression of the gut-homing receptor alpha4beta7 and reduced L-selectin expression, 1,25D(3) had no effect on other homing receptors. In an in vivo assay treatment with RA or 1,25D(3) downregulated the skin infiltration of effector CD4+ T cells. CONCLUSION: These findings suggest that 1,25D(3) can selectively downregulate CLA expression without influencing lymphocyte migration patterns to other tissues. PMID- 17910896 TI - Differential induction of phenylalanine ammonia-lyase gene expression in response to in vitro callus unions of Prunus spp. AB - Phenylalanine ammonia-lyase (PAL) is a key enzyme in the synthesis of phenolic compounds, which play a prominent role in graft union formation, including the marked effects of their accumulation in incompatibility response. The purpose of this study was to assess changes in the abundance of PAL mRNA during graft union development. Partial cDNA clones encoding the enzyme were isolated from in vitro callus tissue in the apricot (Prunus armeniaca L.) cultivar Moniqui and the plum rootstock Marianna 2624 (Prunus munsoniana x Prunus cerasifera). The deduced partial amino acid sequence showed high homology with PAL genes from other plant species. We monitored PAL expression 5, 10, 15 and 20 days after the establishment of in vitro callus unions. The levels of PAL mRNA increased 5 days after grafting in both compatible and incompatible unions. Nevertheless, significant differences were observed at the transcript level through both types of combinations from the second week. The results showed a higher level of PAL transcription in graft unions of incompatible partners, where a lack of adaptation between stock and scion takes place. The level of scion-stock compatibility was related to the PAL expression pattern. In addition, cell walls of the callus cells were not stained by phloroglucinol-HCl, indicating that the increased PAL expression did not result in the formation of lignin. However, staining with Naturstoff reagent A confirmed the highest accumulation of soluble and wall-bound phenolic compounds at the graft interface of incompatible unions. PMID- 17910897 TI - Zinc distribution and zinc-binding forms in Phragmites australis under zinc pollution. AB - The influence of zinc (Zn) on physiological and biochemical parameters was studied to elucidate the mechanism of Zn resistance in Phragmites australis. Zn concentrations in roots, stems and leaves increased with exogenous Zn concentration, while Zn content in roots was much higher than in shoots. X-ray microanalysis was used to reveal compartments in which Zn accumulated in root cortex. Zinc concentrations followed a gradient with the sequence: intercellular space>cell wall >vacuole >cytoplasm, indicating that most Zn was immobilized in the apoplast or sequestered into the vacuolar lumen. Sequential extraction of various Zn chelates revealed that the ratio of Zn extracted with different extraction media was markedly different. Ethanol, HAc (acetic acid) and NaCl extractable Zn were dominant in both roots and leaves of P. australis. Zn-binding protein fractions were found in the roots and leaves after gel filtration chromatography, among which a polypeptide with an apparent molecular mass of 14 kDa bound Zn most effectively. Two newly synthesized polypeptides of 58 and 45 kDa appeared under Zn pollution, whereas a prominent fraction of 72 kDa disappeared. The involvement of Zn distribution in plant tissues, subcellular compartments and chelates and Zn-inducing proteins in the acclimation mechanism of P. australis to Zn pollution is discussed. PMID- 17910898 TI - How the inherent stiffness of the in vivo human trunk varies with changing magnitudes of muscular activation. AB - BACKGROUND: The abdominal muscles provide stiffness to the torso in a manner that is not well understood. Their unique anatomical arrangement may modify their stiffening ability with respect to the more commonly studied long strap-like muscles of the limbs. The purpose of this study was to examine stiffness inherent to the trunk, as modified by different torso, and in particular, abdominal muscle activation levels. METHODS: Nine healthy male participants were secured in a "frictionless" apparatus and subjected to applied bending moments about either the flexion/extension or lateral bend axes. Abdominal muscle activation levels were modified through biofeedback from the right external oblique muscle. Moment angle curves were generated and characterized by an exponential function for each of flexion, extension, and right-side lateral bend, at each of four abdominal muscle activation target level conditions. FINDINGS: Stiffness measured in extension increased in a linear fashion throughout the range of motion and increased with each successive rise in abdominal activation. Stiffness in flexion and lateral bend increased in an exponential fashion over the range of motion. In flexion and lateral bend, stiffness increased with each successive rise in abdominal activation from zero to approximately 40% and 60% of the range of motion, respectively. After these points, stiffness at the highest levels of activation displayed a "yielding" phenomenon whereby the torso stiffness dropped below that characterized at lower levels of activation. INTERPRETATION: Increasing torso muscle co-activation leads to a rise in trunk stiffness over postures most commonly adopted by individuals through daily activities (neutral to approximately 40% of maximum range of motion). However, towards the end range of motion in both flexion and lateral bend, individuals became less stiff at the maximum abdominal muscle co-activation levels. The source and mechanism of this apparent yielding are not fully understood; future work will be directed toward elucidating the cause. PMID- 17910899 TI - Biomechanical comparison of screws and plates for hallux valgus opening-wedge and Ludloff osteotomies. AB - BACKGROUND: The optimal osteotomy type and fixation method for hallux valgus correction have not been defined. This study examined the mechanical properties of corrective opening-wedge and Ludloff oblique osteotomies under conditions approximating postoperative weight-bearing. METHODS: Twenty-nine pairs of fresh frozen metatarsals were divided into three groups. In Group 1, headless screws were compared with standard cortical screws for Ludloff osteotomy fixation. In Groups 2 and 3, Ludloff osteotomies fixed with headless screws were compared with opening-wedge osteotomies fixed with non-locking and locking plates, respectively. Constructs underwent dorsally-directed cantilever loading for 1000 cycles. FINDINGS: No significant differences in angulation or stiffness were demonstrable in Group 1. In Group 2, Ludloff/headless screw construct stiffness exceeded non-locking plate construct stiffness. The mean angulation on the 1000th load cycle was greater for plates than for Ludloff/headless screws. In Group 3, locking plate construct stiffness and angulation did not differ from Ludloff/headless screws in early cyclic loading, but fixation failure of the locking plate constructs was common. INTERPRETATION: The results indicate that screw type for Ludloff fixation may be left to surgeon preference and that opening-wedge plates exhibit mechanical properties inferior to that of the Ludloff osteotomy under the tested conditions. Lateral cortex continuity and bone density remain important factors in the performance of opening-wedge osteotomies. PMID- 17910900 TI - Reflections from an external evaluator on the future of olive production systems on sloping land. PMID- 17910901 TI - Improving public access to environmental information in China. AB - Providing public access to environmental information is a relatively new approach to environmental management that can improve the quality of environmental decision-making and pollution control. China is currently exploring ways to disclose this environmental information, and has found that many aspects of providing environmental information need to be improved. In particular, work still needs to be done to strengthen the laws and regulations that govern public access to environmental information, to expand environmental disclosure among enterprises, to increase disclosure of environmental information in rural areas and enforce the collection of this information, and to investigate and manage environmental information. In this paper, we review the current situation in China, discuss the problems related to this situation, and present recommendations for improvement. PMID- 17910904 TI - Thanks for the memory... PMID- 17910902 TI - Glut-1 antibodies induce growth arrest and apoptosis in human cancer cell lines. AB - Glucose transporters (Gluts) facilitate glucose uptake and tumors frequently over express the Gluts, especially Glut-1. Breast cancer and lung cancer (NSCLC) cell lines were incubated with anti-Glut-1 antibodies alone or with cisplatin, paclitaxel or gefitinib. Inhibition of proliferation and apoptosis was assessed. Antibodies to Glut-1 inhibited proliferation by 50% and 75% in the tested NSCLC and breast cancer cell lines, respectively. They also potentiate the anti proliferative effects of cisplatin, paclitaxel and gefitinib. Our results indicate that anti-Glut-1 antibodies inhibit proliferation and induce apoptosis in the evaluated cell lines and provide preliminary evidence of their anti-tumor activity. PMID- 17910903 TI - Selective stimulation of GalR1 and GalR2 in rat substantia gelatinosa reveals a cellular basis for the anti- and pro-nociceptive actions of galanin. AB - Galanin modulates spinal nociceptive processing by interacting with two receptors, GalR1 and GalR2. The underlying neurophysiological mechanisms were examined by whole-cell recording from identified neurons in the substantia gelatinosa of young adult rats. GalR1 was activated with a 'cocktail' containing the GalR1/2 agonist, AR-M 961 (0.5 microM), in the presence of the GalR2 antagonist, M871 (1.0-2.5 microM). GalR2 was activated with the selective agonist, AR-M 1896 (0.5-1.0 microM). Application of the 'GalR1 agonist cocktail' often activated an inwardly-rectifying conductance in delay firing (excitatory) and tonically firing (inhibitory) neurons. This conductance was not activated by AR-M 1896 which instead decreased or increased an outwardly-rectifying conductance at voltages positive to -70 mV. Despite this variability in its actions on current-voltage relationships, AR-M 1896 very consistently decreased membrane excitability, as measured by cumulative action potential latency in response to a depolarizing current ramp. This strong GalR2-mediated effect was seen in neurons where membrane conductance was decreased, and where membrane excitability might be predicted to increase. GalR2 was also located presynaptically, as AR-M 1896 increased the interevent interval of spontaneous EPSCs in both delay and tonic cells. By contrast, the 'GalR1 agonist cocktail' had little effect on spontaneous EPSCs, suggesting that presynaptic terminals do not express GalR1. These diverse actions of GalR1 and GalR2 activation on both inhibitory and excitatory neurons are discussed in relation to the known spinal antinociceptive and pro-nociceptive actions of galanin, to the possible association of GalR1 with the inhibitory G-protein, G(i/o) and to report that GalR2 activation suppresses Ca2+ channel currents. PMID- 17910905 TI - Pain flashbacks following the July 7th 2005 London bombings. AB - Flashbacks in posttraumatic stress disorder (PTSD) are commonly experienced as visual, auditory, olfactory or tactile re-livings of a previously experienced traumatic event. We present the case report of one survivor of the July 7th 2005 London underground bombings who was diagnosed with PTSD and who experienced painful flashbacks. We present retrospective multidimensional measures of his pain using standardised instruments. The case provides further evidence that somatosensory re-experiencing of pain memories is possible. Findings are discussed with regards to memory for pain. PMID- 17910906 TI - The current status of intensity-modulated radiation therapy in the treatment of nasopharyngeal carcinoma. AB - Radiation therapy is the mainstay of treatment in nasopharyngeal carcinoma, alone or in combination with chemotherapy. In the early stage, it can be managed by radiotherapy alone. For locally advanced-stage disease, several meta-analyses have demonstrated the role of concurrent chemoradiotherapy. Conventional radiation therapy results in significant side effects, particular xerostomia, leading to poor quality of life. With the maturity of intensity-modulated radiation therapy in the recent 10 years, more and more evidences have shown the advantages of intensity-modulated radiation therapy over conventional radiation therapy, regarding the local-regional control, survival rate and quality of life. This article reviews the utilization of intensity-modulated radiation therapy in the management of nasopharyngeal carcinoma with respect to its technical advantages, clinical outcome, critical organ sparing and quality of life, and the dilemma in target delineation. In particular, an issue of treatment-related dysphagia will also be discussed. PMID- 17910907 TI - Androgenic alopecia may have evolved to protect men from prostate cancer by increasing skin exposure to ultraviolet radiation. AB - Androgenic alopecia affects populations adapted to colder climate, and individuals at an age and hormonal status susceptible to prostate cancer. Male pattern baldness enhances absorption of UV radiation on the top of the head, an area directly exposed to sunlight during everyday activities. Ultraviolet radiation is reported to reduce the risk of advanced prostate cancer. Here I propose that progression of androgenic alopecia rather than being a risk factor is a finely tuned mechanism evolved to protect against prostate cancer. PMID- 17910908 TI - Biochemical and cellular mechanisms of aging and degenerative disease: excessive, poor-quality caloric intake may deplete essential nutrients and interfere with cellular processes to produce degenerative damage. AB - It has long been known that caloric restriction retards aging and many degenerative disease processes. Various experiments have confirmed that caloric restriction enables animals' organelles and cells to retain their structural integrity into old age, in contrast to ad libitum counterparts. Calorically restricted animals also maintain greatly enhanced immune and DNA repair systems, handle outside threats-infectious agents, toxins, radiation, extreme temperatures more proficiently, and usually avoid or defer the onset of cancers. But how caloric restriction works to achieve these remarkable results has so far eluded us. Our hypothesis suggests that the key to understanding how caloric and nutrient levels can influence so many physiological functions is to perceive food intake not only as a means of nourishing and strengthening the organism, but also as an environmental challenge. The organism's biochemical responses to excessive, particularly poor-quality, calorie intake, like its responses to other environmental threats, can potentially deplete essential nutrients, interfere with biochemical and cellular mechanisms, and produce degenerative damage. This chain of events occurs not only in animals in caloric restriction experiments, but also in virtually all organisms as they respond to chronic or high-level environmental challenges or receive inadequate nutrition. Because they cannot avoid these challenges, consume precise levels of nutrients required to deal with ongoing metabolic functions and outside threats, and do not function physiologically with total efficiency, all organisms incur degenerative damage, essentially every day, promoting aging and leading to various degenerative disorders. PMID- 17910909 TI - Magnetic lymphatic targeting drug delivery system using carbon nanotubes. AB - By controlling size, nanoparticles can be effectively taken up into lymphatics. On this basis, various nanoparticles have been investigated for transporters of chemotherapeutic pharmaceuticals, but only a few were retained in the draining lymph node. Here, we present a technology using a magnetic carbon nanotubes (MNTs) delivery system, and it may be possible to facilitate the targeted delivery of drugs in the lymphatic tissue more effectively. Chemotherapeutic agents were incorporated into the pores of functionalized MNTs synthesized with a layer of magnetite nanoparticles on the inner surface of the nanotubes. To improve drug delivery to cancer cells in the lymph nodes, individualized MNTs were noncovalently functionalized by folic acid (FA). By using an externally placed magnet to guide the drug matrix to the regional targeted lymph nodes, the MNTs can be retained in the draining targeted lymph nodes for several days and continuously release chemotherapeutic drugs. Selective killing of tumor cells overexpressing the folate receptors (FRs) in the lymph nodes can be achieved, as FR is overexpressed across a broad spectrum of human tumors. PMID- 17910910 TI - Exercise-induced redistribution of T lymphocytes is regulated by adrenergic mechanisms. AB - Acute exercise is known for causing considerable changes in leukocyte counts and function. In this paper we report that differentiated changes in T-lymphocyte distribution occur in lymphoid and non-lymphoid organs depending on the type and the intensity of exercise. Using fluorescent cell tracking we observed a release of T-cells from the spleen while lung, bone marrow and Peyer's patches served as target organs. The number of T-cells in the blood rose after intensive running while lymphopenia occurred after swimming exercise. Changes in number of labelled T-cells were neither found in the lymph nodes nor in the thymus regardless of exercise protocol. Following an alpha- or beta-blockade, the exercise-induced release of T-cells from the spleen and the accumulation of T-cells in the lung were inhibited while the enhancement of T-cells in the Peyer's patches was not affected. The administration of epinephrine partially mimicked the effects of exercise and resulted in a release of T-cells from both, the spleen and the liver, as well as in an increase of circulating blood T-cells. In conclusion, exercise induces a substantial re-distribution of T-cells within lymphoid and non lymphoid organs. The migrating properties of T-cells could be partially explained by adrenergic mechanisms associated with exercise while the involvement of certain homing receptors remains to be shown. Our results suggest that the accumulation of T-cells in both, lung and Peyer's patches, may enhance the immune vigilance in these compartments which serve as the body's major defence barriers. PMID- 17910911 TI - 2- and 3-dimensional echocardiographic analysis of an unusual transient apical ballooning. AB - We report the clinical case of a 60-year-old woman who referred to our hospital for the occurrence of typical chest pain during mild effort. At admission, the electrocardiogram showed S-T segment elevation from V(3) to V(6), and an increase in troponin I level (11.4 ng/mL). Echocardiogram showed midapical segment akinesia with depressed ejection fraction (30%). Basal segments were hypercontractile and there was evidence of dynamic obstruction of the left ventricle with an end-systolic peak gradient of 65 mm Hg. Results of emergency coronary arteriography were normal and left ventricular angiography confirmed the midapical akinesia and hypercontractility of the basal segments. Serial 2- and 3 dimensional Doppler echocardiographic examinations were performed. Regression of left ventricular outflow tract obstruction was soon detected (day 3). Fifteen days after admission, 2- and 3-dimensional echocardiography showed a complete regression of both apical ballooning and wall-motion abnormalities with an improvement in overall systolic function. Segmental volumetric analysis allowed accurate assessment of regional volumes and ejection fraction, which were indicative for a progressive reverse remodeling. Regression of wall-motion abnormalities was expressed by a normalization in regional ejection fraction curves at 15 days. PMID- 17910912 TI - Application of sewage sludge compost on highway embankments. AB - More and more sewage sludge is being produced in China. Safe and economical methods for sewage sludge disposal should be found considering the increase in sewage treatment. In order to verify the feasibility of sludge disposal on newly built highway embankments, five treatments (0, 15, 30, 60 and 120 tons ha(-1)) of sewage sludge compost (SSC) were added to a silty-clay embankment soil on the Xi Huang highway. The results showed that amendment with SSC increased soil available N, available P, organic matter, cation exchange capacity, and water content, and decreased soil bulk density. Application of SSC enhanced ryegrass growth and reduced runoff and soil erosion. Heavy metal losses from sediments in runoff remained constant or decreased relative to the control until a rate of 60 tons ha(-1) was exceeded, when heavy metal losses appeared to increase. PMID- 17910913 TI - Reuse of thermosetting plastic waste for lightweight concrete. AB - This paper presents the utilization of thermosetting plastic as an admixture in the mix proportion of lightweight concrete. Since this type of plastic cannot be melted in the recycling process, its waste is expected to be more valuable by using as an admixture for the production of non-structural lightweight concrete. Experimental tests for the variation of mix proportion were carried out to determine the suitable proportion to achieve the required properties of lightweight concrete, which are: low dry density and acceptable compressive strength. The mix design in this research is the proportion of plastic, sand, water-cement ratio, aluminum powder, and lignite fly ash. The experimental results show that the plastic not only leads to a low dry density concrete, but also a low strength. It was found that the ratio of cement, sand, fly ash, and plastic equal to 1.0:0.8:0.3:0.9 is an appropriate mix proportion. The results of compressive strength and dry density are 4.14N/mm2 and 1395 kg/m3, respectively. This type of concrete meets most of the requirements for non-load-bearing lightweight concrete according to ASTM C129 Type II standard. PMID- 17910914 TI - The cost-effectiveness of bevacizumab in the first-line treatment of metastatic colorectal cancer in England and Wales. AB - BACKGROUND: Bevacizumab is a humanised monoclonal antibody, which has demonstrated significant activity in metastatic colorectal cancer. The aim of this study is to estimate the cost-effectiveness of adding bevacizumab to chemotherapy for patients with untreated metastatic colorectal cancer. METHODS: A decision-analytic model was developed to estimate the lifetime costs and benefits of adding bevacizumab to irinotecan plus FU/LV (IFL) or 5-FU/LV alone. Effectiveness outcomes, health utilities and resource use data were derived from recent bevacizumab RCTs and from the literature. RESULTS: Adding bevacizumab to IFL costs approximately pound62,857 per QALY gained. Adding bevacizumab to 5 FU/LV costs approximately pound88,436 per QALY gained. The acquisition cost of bevacizumab is a key determinant of its cost-effectiveness. The probability that bevacizumab has a cost-effectiveness ratio that is better than pound30,000 per QALY gained is close to zero. CONCLUSIONS: Given high acquisition costs in relation to clinical benefits, bevacizumab is unlikely to represent a cost effective use of NHS resources. PMID- 17910915 TI - Biphenyls as potent vitronectin receptor antagonists. Part 3: Squaric acid amides. AB - Vitronectin receptor (alpha(V)beta(3)) antagonists have been implicated as a possible new treatment of restenosis following balloon angioplasty. In this work we investigate a series of novel arginine mimetic scaffolds leading to new insight of the alpha(V)beta(3)/ligand interaction. Squaric acid amide 10 is a subnanomolar alpha(V)beta(3) antagonist with improved potency on human smooth muscle cell migration. PMID- 17910916 TI - Synthesis and structure-activity relationships of 16-modified analogs of 2 methoxyestradiol. AB - A series of 16-modified 2-methoxyestradiol analogs were synthesized and evaluated for antiproliferative activity toward HUVEC and MDA-MB-231 cells, and for susceptibility to conjugation. In addition, the estrogenicity of these analogs was accessed by measuring cell proliferation of the estrogen-dependent cell line MCF7 in response to compound treatment. It was observed that antiproliferative activity dropped as the size of the 16 substituent increased. Selected analogs tested in glucuronidation assays had similar rates of clearance to 2 methoxyestradiol, but had enhanced clearance in sulfonate conjugation assays. PMID- 17910917 TI - Comment on "Sequential determination of Pu and Am radioisotopes in environmental samples: A comparison of two separation procedures" by R. Jakopic, P. Tavcar and L. Benedik, Appl. Radiat. Isot. 65 (2007) 504-511. PMID- 17910918 TI - Multivariate statistical analyses of artificial radionuclides and heavy metals contaminations in deep mud of Keban Dam Lake, Turkey. AB - On the basis of the artificial radioactive and heavy metal compositions, factor and cluster analyses are employed to identify the inter-relationship among different variables and their similarity groups. In this paper, 15 physico chemical variables, including the activities of 137Cs, 90Sr, total alpha, total beta, and concentrations of Fe, Mg, Ca, K, Na, Zn, Cu, Cr, Co, Ni, and Mn, are used for the application of the proposed methodologies. The spatio-temporal samples of these variables are collected from deep mud at about 30-35 m mean depth in the reservoir of Keban Dam Lake, which is located in the eastern part of Turkey. Spatially, there are 20 sampling sites at the dam with 150 km2 lake surface area where samples were taken in 2006. The lake is affected by man-made and industrial influxes. The application of the factor and cluster analysis methods yields that the former method reduces the number of variables into six factors with 77.2% variance explanation whereas the latter yields three distinctive groups of the same variables. PMID- 17910919 TI - Cloning and expression pattern of dog SDF-1 and the implications of altered expression of SDF-1 in ischemic myocardium. AB - Stromal cell derived factor-1 (SDF-1) plays a pivotal role in the mobilization and homing of stem cells, indicative of its potential in myocardial regeneration after heart damage. The use of large animal models in cardiac surgery research plays an essential role in the translation of results from basic studies into clinical trials. Considering the aforesaid two reasons, for the first time, we cloned dog SDF-1 cDNA and characterized the constitutive expression pattern of SDF-1 gene in normal dog tissues and the dynamic expression pattern in a dog model of myocardial infarction (MI) by means of ELISA test, real-time RT-PCR, and Western blotting. In a dog model of MI, We also examined and compared the differentially expressed pattern of SDF-1 between infarct area and border zone of myocardium in order to explore the implication of differentially distribution of SDF-1 in the mobilization and homing of stem cells to the damaged heart. Our results provide insights into expression pattern and pathophysiologic significance of dog SDF-1 in normal and heart-damaged dogs. PMID- 17910920 TI - Expression and bioactivity analysis of Staphylococcal enterotoxin M and N. AB - Staphylococcal enterotoxins (SEs) are powerful superantigens that stimulate non specific T-cell proliferation produced by Staphylococcus aureus and draw considerable attention as ideal drugs for cancer therapy. The filtrate of S. aureus culture has been used as ampul named Staphylococcal enterotoxin C injection in clinic for 10 years in China and proved to be effective. The superantigen SEC claimed to be the only active component without certifiable evidences. For further investigations of the active components of this injection and establishment of foundations for the development of novel anti-cancer drugs, in this research we extracted total DNA from S. aureus (FRI 1230), cloned, expressed and purified recombinant proteins of Staphylococcal enterotoxin M and N (rSEM and rSEN). The MTT assay of the purified rSEM and rSEN demonstrated that their abilities of stimulating T cells and inhibiting the proliferation of K562 ADM cells and B16 cells were equivalent to that of purified SEC2 in vitro. These findings suggested that SEC was not the only active component of Staphylococcal enterotoxin C injection and the effective procedure of expression and purification may be useful for mass productions of these therapeutically important proteins. PMID- 17910921 TI - Biophysical and biochemical characterization of reconstituted and purified Rhodobacter sphaeroides cytochrome c oxidase in phospholipid vesicles sheds insight into its functional oligomeric structure. AB - Discontinuous sucrose gradient ultracentrifugation was used to separate liposomes containing Rhodobacter sphaeroides cytochrome c oxidase (pCOV) from liposomes devoid of the enzyme, and the biophysical and biochemical properties of pCOV were compared to unpurified liposomes containing cytochrome c oxidase (COV). Isolated and purified R. sphaeroides cytochrome c oxidase (COX) was reconstituted into asolectin phospholipid vesicles by cholate dialysis, and this preparation was purified further on a discontinuous sucrose gradient to isolate only those vesicles which contained the enzyme (pCOV). After centrifugation at 300,000g for 22h, 80% of the enzyme recovered was in a single band. The number of COX molecules per pCOV liposome was estimated by measuring the visible absorbance spectrum of cytochrome c oxidase (for heme aa(3)) and inorganic phosphate concentration (for phospholipid). The number of COX molecules incorporated per pCOV was estimated to be approximately one (0.72+/-0.19-1.09+/-0.28). The pCOV exhibited similar physical properties as COV; respiratory control ratios (indicators of endogenous proton permeability) and maximum enzymatic turnover number at pH 7.4 were comparable (6.0+/-1.3 and 535+/-130s(-1)). Furthermore, proton pumping activities of the pCOV were at least 70% of COV, indicating that discontinuous sucrose gradient centrifugation is a useful technique for functional experiments in R. sphaeroides cytochrome c oxidase. Our results suggest that the monomeric form of R. sphaeroides COX when reconstituted into a phospholipid bilayer is completely functionally active in its ability to perform electron transfer and proton pumping activities of the enzyme. PMID- 17910922 TI - A highly efficient integrated chromatographic procedure for the purification of recombinant hepatitis B surface antigen from Hansenula polymorpha. AB - The high expression level of recombinant hepatitis B surface antigen obtained from Hansenula polymorpha yeast cell (Hans-HBsAg) made it possible to produce HBsAg vaccine in a large scale and by cost-effective process. However, the present available purification process was somewhat tedious, time-consuming and difficult to scale up. To improve the purification efficiency and simplify the purification process, an integrated chromatographic process was developed and optimized. The downstream process included ion-exchange chromatography (IEC), hydrophobic interaction chromatography (HIC) and gel filtration chromatography (GFC). A series of chromatographic adsorbents were evaluated for their performances on the purification of Hans-HBsAg, and then the suitable adsorbents for IEC and HIC were screened out, respectively. After clarification by centrifugation, the supernatant of cell disruption (SCD) was purified by standard chromatographic steps, IEC on DEAE Sepharose FF, HIC on Butyl-S-QZT and GFC on Sepharose 4FF. Furthermore, HBsAg recovery, purification factor (PF) and purity during the downstream process were evaluated with enzyme-linked immunosorption assay (ELISA), sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS PAGE) and high-performance size-exclusion chromatography (HPSEC). The results demonstrated that in the scale of 550ml SCD, the total HBsAg recovery and PF of the whole procedure were about 21.0+/-0.9% and 80.7+/-8.4 (n=3) respectively, with the purity of above 99%. This new downstream process was efficient, reproducible and relatively easy to be scaled up. PMID- 17910923 TI - Molecular systematics of the barklouse family Psocidae (Insecta: Psocodea: 'Psocoptera') and implications for morphological and behavioral evolution. AB - We evaluated the higher level classification within the family Psocidae (Insecta: Psocodea: 'Psocoptera') based on combined analyses of nuclear 18S, Histone 3, wingless and mitochondrial 12S, 16S and COI gene sequences. Various analyses (inclusion/exclusion of incomplete taxa and/or rapidly evolving genes, data partitioning, and analytical method selection) all provided similar results, which were generally concordant with relationships inferred using morphological observations. Based on the phylogenetic trees estimated for Psocidae, we propose a revised higher level classification of this family, although uncertainty still exists regarding some aspects of this classification. This classification includes a basal division into two subfamilies, 'Amphigerontiinae' (possibly paraphyletic) and Psocinae. The Amphigerontiinae is divided into the tribes Kaindipsocini (new tribe), Blastini, Amphigerontini, and Stylatopsocini. Psocinae is divided into the tribes 'Ptyctini' (probably paraphyletic), Psocini, Atrichadenotecnini (new tribe), Sigmatoneurini, Metylophorini, and Thyrsophorini (the latter includes the taxon previously recognized as Cerastipsocini). We examined the evolution of symmetric/asymmetric male genitalia over this tree and found this character to be quite homoplasious. PMID- 17910924 TI - Antimicrobial activity and protective properties of vaginal lactobacilli from healthy Bulgarian women. AB - The role of vaginal Lactobacillus as an efficient barrier against invading pathogens is of considerable interest. The purpose of the present study was to assess in vitro the ability of 20 recently identified vaginal lactobacilli to protect the vagina. In order to evaluate their significance, the antimicrobial, hemagglutination (HA) and aggregation (Agg) activities, as well as acid and hydrogen peroxide (H(2)O(2)) production, were estimated. The cell-free cultures of eight strains showed a stable antimicrobial activity after elimination of the putative effects of lactic acid and H(2)O(2). Three of the isolated vaginal lactobacilli expressed a broad spectrum of anti-bacterial activity including Gram negative pathogens. Strains with anti-Gardnerella and anti-herpes simplex virus type 2 activities were found. All tested isolates were H(2)O(2) producers, actively acidifying the growth media to pH 3.92+/-0.04, which is presumed to neutralize sexually transmitted infection pathogens. The major part (75%) expressed an HA activity and different Agg phenotypes, estimated as important properties in the competition with invading pathogens and in host defense. These results are encouraging and prompt further research of the characterized active strains and their possible application in prophylaxis of vaginal disorders. PMID- 17910925 TI - Inherited abnormalities of skeletal development in sheep. AB - Inherited diseases of the skeleton are reported less often in sheep than in most other domestic animal species but are likely to occur more frequently than the veterinary literature would suggest. Although most are lethal or semi-lethal, the gene frequency for some of these diseases has reached surprisingly high levels in defined populations, presumably due either to the founder effect or the presence of a selective advantage of heterozygous individuals. This article reviews the clinical characteristics, pathology, mode of inheritance and molecular basis of skeletal diseases known to have a genetic aetiology in sheep. Inherited skeletal diseases of sheep are potential models for studying the treatment of similar diseases in humans. PMID- 17910926 TI - Ganglioneuroblastoma in a cat: a rare neoplasm treated with electrochemotherapy. AB - An 8-year-old male castrated cat was referred for sudden onset of lameness. Physical examination revealed a 1x2x1cm mass originating from a footpad of the right hind leg. A diagnosis of ganglioneuroblastoma was suggested by the tumour appearance following histopathological staining with haematoxylin and eosin and haematoxylin/van Gieson. Immunohistochemical staining for glial fibrillary acidic protein (GFAP), vimentin, neuron-specific enolase (NSE), neurofilament and S100 further confirmed the diagnosis. The staging process did not indicate metastatic spread. The cat was treated with three sessions of electrochemotherapy (ECT) 1 week apart, following local injection of bleomycin. The tumour had completely regressed within 1 week of the third ECT application and remained in remission for 402 days at which time a small recurrence was noted. The animal was given a further session of ECT using intra-lesional cisplatin and again went into remission. It remained tumour free at 450 days. Electrochemotherapy is considered a safe and effective treatment for localised neoplasms of cats and dogs and warrants further investigation. PMID- 17910927 TI - A Hall effect angle detector for solid-state NMR. AB - We describe a new method for independent monitoring of the angle between the spinning axis and the magnetic field in solid-state NMR. A Hall effect magnetic flux sensor is fixed to the spinning housing, so that a change in the stator orientation leads to a change in the angle between the Hall plane and the static magnetic field. This leads to a change in the Hall voltage generated by the sensor when an electric current is passed through it. The Hall voltage may be measured externally by a precision voltmeter, allowing the spinning angle to be measured non-mechanically and independent of the NMR experiment. If the Hall sensor is mounted so that the magnetic field is approximately parallel to the Hall plane, the Hall voltage becomes highly sensitive to the stator orientation. The current angular accuracy is around 10 millidegrees. The precautions needed to achieve higher angular accuracy are described. PMID- 17910929 TI - Quantitative evaluation of the use of microbubbles with transcranial focused ultrasound on blood-brain-barrier disruption. AB - It has been shown that focused ultrasound (FUS) can disrupt the blood-brain barrier (BBB) noninvasively and reversibly at target locations when applied in the presence of ultrasound contrast agent (UCA). In this study, the dose dependent effects of UCA on BBB disruption were investigated in the brains of 16 male Wistar rats sonicated by 1.0-MHz transcranial FUS, with the UCA present at four doses. The BBB disruption was evaluated quantitatively based on the extravasation of Evans blue (EB). The amount of EB extravasation in the brain increased with the quantity of UCA injected into the femoral vein prior to sonication. Moreover, the use of a suitable dose of UCA resulted in the BBB disruption being concentrated in the focal region instead of the entire brain. Our results indicate that injecting an appropriate quantity of UCA effectively increases and localizes the BBB disruption induced by transcranial FUS sonications. PMID- 17910928 TI - The effect of gestational age and labor on placental growth hormone in amniotic fluid. AB - OBJECTIVE: Placental growth hormone (PGH) is produced by trophoblast. This hormone becomes detectable in maternal serum during the first trimester of pregnancy. Its concentration increases as term approaches and becomes undetectable within one hour of delivery. PGH has important biological properties, including somatogenic (growth promotion), lactogenic, and lipolytic activity. Recently, PGH has been detected in amniotic fluid (AF) of midtrimester pregnancies. The purpose of this study was to determine whether PGH concentrations in AF change with advancing gestational age and in labor at term. DESIGN: AF was assayed for PGH concentrations in samples obtained from patients undergoing genetic amniocentesis between 14 and 18 weeks of gestation (n=67), normal patients at term not in labor (n=24), and pregnant women at term in labor (n=51). PGH concentrations were determined by ELISA. Non-parametric statistics were used for analysis. RESULTS: (1) PGH was detected in all AF samples; (2) patients in the midtrimester had a higher median concentration of PGH in AF than those at term (midtrimester: median: 3140.5 pg/ml; range: 1124.2-13886.5 vs. term: median: 2021.1pg/ml; range: 181.6-8640.8; p<0.01); (3) there was no difference in the median concentration of PGH between women at term, not in labor, and those in labor (term not in labor: median: 2113.4pg/ml; range: 449.3 8640.8 vs. term in labor: median: 2004.1pg/ml; range: 181.6-8531.5; p=0.73). CONCLUSIONS: (1) PGH is detectable in AF at both mid- and third trimesters; (2) the median AF concentration of PGH is significantly lower at term when compared to the second trimester; (3) labor at term is not associated with changes in the AF concentration of PGH. The role of this unique placental hormone now found in the fetal compartment requires further investigation. PMID- 17910930 TI - Motion analysis study of a scapular orientation exercise and subjects' ability to learn the exercise. AB - Exercises to retrain the orientation of the scapula are often used by physiotherapists to optimise shoulder girdle function. The movements and muscle activity required to assume this position have not yet been quantified. Further, patients often find this a difficult exercise to learn accurately, with no data being available on the accuracy of repeated performance. The primary objective of this study was to quantify the movements occurring during a commonly used scapular orientation exercise. The secondary objective was to describe the ability of subjects to learn this position after a brief period of instruction. A group of normal subjects (13 subjects; mean age 32, SD=9) were taught the scapular orientation exercise. Measurement of the position and muscle actions were made with a motion analysis system and surface electromyography. Further comparison was made of the accuracy of repeated trials. The most consistent movements were upward (mean=4 degrees, SEM=0.9 degrees) and posterior rotation (mean=4 degrees, SEM=1.6 degrees). All parts of the trapezius muscle demonstrated significant activity in maintaining the position while latissimus dorsi did not. Repeated trials showed that subjects were able to accurately repeat the movement without guidance. The key movements of, and immediate efficacy of a teaching approach for, scapular orientation have been established. PMID- 17910931 TI - Measuring the posteroanterior stiffness of the cervical spine. AB - An essential part of improving manual therapy treatment for cervical spine disorders is the identification of the mechanical effects of manual techniques. The aims of this research were to develop a reliable and safe instrument for measuring cervical spine stiffness, and to document stiffness in a group of asymptomatic individuals. A device for measuring cervical spine stiffness was designed and tested. The stiffness of the cervical spine of 67 asymptomatic individuals was measured at C2 and C7 on one or more occasions. Stiffness was defined as the slope of the linear region of the force-displacement curve (coefficient K). For C2, the linear region of the force-displacement curve was from 7 to 40 N, and for C7, 20-70 N. The mean stiffness (coefficient K) on the first measurement occasion at C2 was 4.58 N/mm (95% CI 4.30-4.85), and at C7 was 7.03 N/mm (95% CI 6.50-7.57). ICC(2,1) for repeated measurements was 0.84 (95% CI 0.74-0.90). Stiffness measurements in the cervical spine were generally lower than those previously reported for the lumbar spine. Age was positively associated with C2 stiffness (p=0.01). Males were stiffer at C7 than females (p<0.001). This research provides a basis for future studies investigating the effects of manual techniques on cervical spine stiffness, potentially leading to improved outcomes for patients treated by manual therapy. PMID- 17910932 TI - Bilateral and unilateral increases in calcaneal eversion affect pelvic alignment in standing position. AB - Excessive foot pronation has been associated with the occurrence of low back pain, possibly for generating changes in the lumbopelvic alignment. However, the influence of foot pronation (measured as calcaneal eversion) on pelvic alignment during standing has not been well established. Fourteen young healthy subjects participated in the study. A Motion Analysis System was used to obtain pelvic positions in sagittal and frontal planes and calcaneal position in the frontal plane. Volunteers were filmed in relaxed standing position during three trials, in three conditions: control; unilateral experimental with increased right calcaneal eversion and bilateral experimental with increased bilateral calcaneal eversion. Increased calcaneal eversion was obtained using wedges tilted 10 degrees medially, unilaterally and bilaterally. Repeated measures ANOVAs with Bonferroni corrections were used for statistical analysis. Unilateral and bilateral use of medially tilted wedges produced a significant increase of calcaneal eversion (P3)-alpha-l rhamnopyranosyl(1-->2)-alpha-l-arabinopyranoside (OA) from Aralia elata inhibits LPS-induced nitric oxide production by down-regulated NF-kappaB in raw 264.7 cells. AB - It is well known that the pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin (PG)E(2) are involved in several inflammatory diseases and lipopolysaccharide (LPS) can stimulate these inflammatory responses. Oleanolic acid 3-O-beta-d-glucopyranosyl(1-->3)-alpha-l-rhamnopyranosyl(1-->2)-alpha-l arabinopyranoside (OA) was purified from edible plant Aralia elata. OA inhibited LPS-induced NO and PGE(2) production in raw 264.7 murine macrophages in a dose dependent manner and RT-PCR analysis indicated OA inhibited mRNA transcriptions of iNOS and COX-2 genes in LPS-induced cells. EMSA and Western blot analysis revealed that OA drastically reduced NF-kappaB translocation by the inhibition effects of LPS-induced phosphorylation of IkappaBalpha. In addition, it was found that OA inhibited the phosphorylation of ERK1/2, p38 and JNK MAPK, and the treatment of U0126 in LPS-induced raw 264.7 cells showed significant inhibition activity on the NO production and the phosphorylation of IkappaBalpha. Taken together, it is suggested that OA from A. elata has an anti-inflammatory activity via down-regulation of NF-kappaB. PMID- 17910943 TI - Mapping folding energy landscapes with theory and experiment. AB - The detailed characterization of the overall free energy landscape associated with the folding process of a protein is the ultimate goal in protein folding studies. Modern experimental techniques and all-atom simulations provide a way to obtain accurate thermodynamic and kinetic measurements, but they are oftentimes restricted to probe limited regions of a protein landscape. Although simplified protein models can access larger regions of the landscape, they are built on assumptions and approximations that can affect the accuracy of the results. We review here recent promising approaches that allow to combine the complementary strengths of theory and experiment for a more complete characterization of a protein folding landscape at multiple resolutions. Recent results and possible applications are discussed. PMID- 17910944 TI - Brain expression of the calcineurin inhibitor RCAN1 (Adapt78). AB - RCAN1 (Adapt78) is an endogenous inhibitor of calcineurin, an important intracellular phosphatase that mediates many cellular responses to calcium. RCAN1 is expressed in multiple organs, especially heart, skeletal muscle and brain. In brain, it is thought to be important due to its strong expression, developmental regulation, abundance of target protein (calcineurin), and putative links to multiple brain-related disorders. Surprisingly, however, few studies have examined RCAN1 protein expression here. This has led to some confusion in the field over the exact nature and cell-type expression of isoform 4, the more studied of the two major RCAN1 protein isoforms, in brain. Here we characterize RCAN1 brain isoforms in more detail by assessing their size and distribution under conditions of calcium elevation, a hallmark of the isoform 4 response, and using rodent models to allow for more expanded analyses. We find that the 25 29kDa version of this protein, reported in many non-brain studies, is indeed also present in neurons, and most observable after calcium induction. We also observe that expression of isoform 4 is not specific to neurons, as both microglia and astrocyte cells in culture exhibit a strong induction of isoform 4 protein following calcium stress that is not observable in non-stressed tissue sections. Isoform 1 expression is also observable in a primary glial cell-type (rat microglia). Finally, our observations confirm previous reports of low or non detectable constitutive isoform expression in non-stressed glia, and of a larger sized, RCAN1 antibody-interacting species. These studies extend and complement previous studies on RCAN isoforms toward better understanding the role of RCAN1 in brain function and as a potential new target for treating calcineurin-related brain disorders. PMID- 17910945 TI - Regulation of SOX3 gene expression is driven by multiple NF-Y binding elements. AB - The presented results demonstrate that human SOX3 promoter possesses three CCAAT box control elements involved in the regulation of SOX3 gene expression in NT2/D1 cells. By mutational analysis we have shown that all three elements are of functional relevance for constitutive SOX3 expression. Electrophoretic mobility shift assays indicate that the active complexes at three sites involve the ubiquitously expressed CCAAT binding protein NF-Y. The involvement of NF-Y in the up-regulation of SOX3 expression in NT2/D1 cells was demonstrated in vivo by Northern and Western blot analyses. Furthermore, in co-transfection experiments we have shown that NF-Y mediates transcriptional activation of SOX3 promoter. Our data indicate that multiple CCAAT control elements are involved in the regulation of the SOX3 promoter, suggesting that NF-Y functions as a key regulator of SOX3 gene expression. Further, our results indicate that these elements can be recognized as modulators of retinoic acid induced activation of SOX3 expression. PMID- 17910946 TI - The vaccination potential of EMY162 antigen against Echinococcus multilocularis infection. AB - Alveolar echinococcosis is caused by infection with the larval stage of Echinococcus multilocularis. We recently identified a cDNA clone, designated as emy162, that encodes a putative secreted protein. EMY162 shares structural features with the EM95 antigen, which is a host-protective antigen. The amino acid sequence of EMY162 shows 31.4% identity to EM95 whereas these antigens are distinguishable with respect to their predicted secondary structure and antigenicity on Western blot analysis. RT-PCR analysis revealed that the gene expression of emy162 was significantly higher than that of em95 at each life cycle stage. Recombinant EMY162 antigen induced a significant level of host protection (74.3%) in experimental infection with E. multilocularis eggs in mice. Notably, recombinant EMY162 antigen showed significant reactivity to the sera from alveolar echinococcosis patients. These results may help in the development of a practical vaccine to reduce the level of alveolar echinococcosis in humans. PMID- 17910947 TI - Identification of twinfilin-2 as a factor involved in neurite outgrowth by RNAi based screen. AB - Using RNA interference (RNAi) to suppress gene expression, we attempted to identify tyrosine kinases involved in the extension of neurites from SH-SY5Y cells. A comprehensive analysis of gene "knock-down" profiles with small interfering RNAs (siRNAs) revealed candidate proteins that might control neurite extension. Phenotype-based screening of differentiating SH-SY5Y cells following retinoic acid (RA) stimulation indicated that twinfilin-2 is a protein that is involved in neurite outgrowth, as confirmed by morphological analysis of twinfilin-2-overexpressing cells. PMID- 17910948 TI - ZNF307, a novel zinc finger gene suppresses p53 and p21 pathway. AB - We have cloned a novel KRAB-related zinc finger gene, ZNF307, encoding a protein of 545aa. ZNF307 is conserved across species in evolution and is differentially expressed in human adult and fetal tissues. The fusion protein of EGFP-ZNF307 localizes in the nucleus. Transcriptional activity assays show ZNF307 suppresses transcriptional activity of L8G5-luciferase. Overexpressing ZNF307 in different cell lines also inhibits the transcriptional activities of p53 and p21. Moreover, ZNF307 works by reducing the p53 protein level and p53 protein reduction is achieved by increasing transcription of MDM2 and EP300. ZNF307 might suppress p53 p21 pathway through activating MDM2 and EP300 expression and inducing p53 degradation. PMID- 17910949 TI - Characterization of the promoter of human CRTh2, a prostaglandin D2 receptor. AB - Chemoattractant-receptor homologous molecule expressed on Th2 cells (CRTh2) is a receptor for prostaglandin (PG)D2, a lipid mediator involved in allergic inflammation. CRTh2 is expressed by Th2 cells, eosinophils and basophils and PDG(2)-CRTh2 signaling induces calcium mobilization, cell migration and expression of the Th2 cytokines IL-4, IL-5, and IL-13. Despite the role of CRTh2 in allergic inflammation, transcriptional regulation of this gene has not been studied. Here, we demonstrated that a reporter construct of the CRTh2 promoter was induced following T cell stimulation. This activity could be further enhanced by over-expression of GATA-3, but not NFAT2 or STAT6. Electromobility shift assay demonstrated GATA-3 binding to a probe from the CRTh2 promoter. This study provides the first detailed analysis of transcriptional regulation of the human CRTh2 promoter. These findings may help identify strategies to attenuate expression of this gene and influence the maintenance and proliferation of Th2 cells in allergic inflammation. PMID- 17910950 TI - Peroxynitrite-mediated oxidation of ferrous carbonylated myoglobin is limited by carbon monoxide dissociation. AB - Peroxynitrite-mediated oxidation of ferrous nitrosylated myoglobin (Mb(II)-NO) involves the transient ferric nitrosylated species (Mb(III)-NO), followed by ()NO dissociation and formation of ferric myoglobin (Mb(III)). In contrast, peroxynitrite-mediated oxidation of ferrous oxygenated myoglobin (Mb(II)-O2) involves the transient ferrous deoxygenated and ferryl derivatives (Mb(II) and Mb(IV)O, respectively), followed by Mb(III) formation. Here, kinetics of peroxynitrite-mediated oxidation of ferrous carbonylated horse heart myoglobin (Mb(II)-CO) is reported. Values of the first-order rate constant for peroxynitrite-mediated oxidation of Mb(II)-CO (i.e., for Mb(III) formation) and of the first-order rate constant for CO dissociation from Mb(II)-CO (i.e., for Mb(II) formation) are h=(1.2+/-0.2)x10(-2)s(-1) and k(off(CO))=(1.4+/-0.2)x10( 2)s(-1), respectively, at pH 7.2 and 20.0 degrees C. The coincidence of values of h and k(off(CO)) indicates that CO dissociation represents the rate limiting step of peroxynitrite-mediated oxidation of Mb(II)-CO. PMID- 17910951 TI - Developmental expression of Commd1 in the liver of the Jackson toxic milk mouse. AB - Wilson disease (WD) is due to mutations in ATP7B, which encodes an intracellular metal-transporting P-type ATPase. In WD holoceruloplasmin production and biliary excretion of copper are decreased, leading to copper overload, oxidative stress and apoptotic cell death. Other copper-binding proteins include COMMD1, which is inactive in the Bedlington terrier hereditary copper toxicosis, and XIAP, which regulates apoptosis. We examined developmental expression of Commd1 and Xiap in the Jackson toxic milk mouse (Atp7b(tx-J), G712D missense mutation in Atp7b). Expression of Commd1 mRNA appeared unchanged by PCR but real-time PCR demonstrated 3- to 4-fold increase over the first 6 months of life. Immunodetectable Xiap dropped over the first 8 months of life and was nearly undetectable from 6 months onward. Cytosolic NF-kappaB rose then dropped precipitously at 5-6 months. In tx-j mice hepatic copper accumulation leads to decreased Xiap, increased Commd1; these responses ultimately fail to prevent progressive apoptotic cell damage. PMID- 17910952 TI - EGFR and beta1 integrins utilize different signaling pathways to activate Akt. AB - Akt, also called PKB, is a serine/threonine kinase that plays a major role in cell survival. It can be activated by several cellular receptors, including integrins and growth factor receptors, in PI3K-dependent manners. In this study, we analyzed the two current models for Akt activation upon beta1 integrin mediated adhesion: via focal adhesion kinase and via transactivation of the EGF receptor. Distinct differences in the pathways leading to phosphorylation and activation of Akt from stimulated beta1 integrins and EGF receptor were observed, including opposing sensitivity to the tyrosine kinase inhibitors PP2 and Gefitinib. Using knockout cells and integrin mutant cells, we show that beta1 integrins can induce phosphorylation of Akt at Ser473 and Thr308 and Akt kinase activity independently of the EGF receptor activity, focal adhesion kinase, and the Src family members. In contrast to stimulation with EGF, beta1 integrin mediated adhesion did not induce Akt tyrosine phosphorylation. Moreover, tyrosine phosphorylation of Akt was found not to be required for its catalytic activity. The results identify a previously unrecognized mechanism by which beta1 integrins activate the PI3K/Akt pathway. PMID- 17910954 TI - Protective effects of metallothionein against dopamine quinone-induced dopaminergic neurotoxicity. AB - Dopamine (DA) quinone as DA neuron-specific oxidative stress conjugates with cysteine residues in functional proteins to form quinoproteins. Here, we examined the effects of cysteine-rich metal-binding proteins, metallothionein (MT)-1 and 2, on DA quinone-induced neurotoxicity. MT quenched DA semiquinones in vitro. In dopaminergic cells, DA exposure increased quinoproteins and decreased cell viability; these were ameliorated by pretreatment with MT-inducer zinc. Repeated L-DOPA administration markedly elevated striatal quinoprotein levels and reduced the DA nerve terminals specifically on the lesioned side in MT-knockout parkinsonian mice, but not in wild-type mice. Our results suggested that intrinsic MT protects against L-DOPA-induced DA quinone neurotoxicity in parkinsonian mice by its quinone-quenching property. PMID- 17910953 TI - Telomere length in Trypanosoma brucei. AB - Trypanosoma brucei thwarts the host immune response by replacing its variant surface glycoprotein (VSG). The actively transcribed VSG is located in one of approximately 20 telomeric expression sites (ES). Antigenic variation can occur by transcriptional switching, reciprocal translocations, or duplicative gene conversion events among ES or with the large repertoire of telomeric and non telomeric VSG. In recently isolated strains, duplicative gene conversion occurs at a high frequency and predominates, but the switching frequency decreases dramatically upon laboratory-adaptation. Uniquely, T. brucei telomeres grow- apparently indefinitely--at a steady rate of 6-12 base pairs (bp) per population doubling (PD), but the telomere adjacent to an active ES undergoes frequent truncations. Using two-dimensional gel electrophoresis, we demonstrate that all of the chromosome classes of fast-switching and minimally propagated T. brucei have shorter telomeres than extensively propagated Lister 427 clones, suggesting a link between laboratory adaptation, telomere growth, and VSG switching rates. PMID- 17910955 TI - Induction of DAN/TIR yeast cell wall mannoprotein genes in response to high hydrostatic pressure and low temperature. AB - Global transcriptional profiles of Saccharomyces cerevisiae were studied following changes in growth conditions to high hydrostatic pressure and low temperature. These profiles were quantitatively very similar, encompassing 561 co upregulated genes and 161 co-downregulated genes. In particular, expression of the DAN/TIR cell wall mannoprotein genes, which are generally expressed under hypoxia, were markedly upregulated by high pressure and low temperature, suggesting the overlapping regulatory networks of transcription. In support of the role of mannoproteins in cell wall integrity, cells acquired resistance against treatment with SDS, Zymolyase and lethal levels of high pressure when preincubated under high pressure and low temperature. PMID- 17910956 TI - Detection of initiation sites in protein folding of the four helix bundle ACBP by chemical shift analysis. AB - A simple alternative method for obtaining "random coil" chemical shifts by intrinsic referencing using the protein's own peptide sequence is presented. These intrinsic random coil backbone shifts were then used to calculate secondary chemical shifts, that provide important information on the residual secondary structure elements in the acid-denatured state of an acyl-coenzyme A binding protein. This method reveals a clear correlation between the carbon secondary chemical shifts and the amide secondary chemical shifts 3-5 residues away in the primary sequence. These findings strongly suggest transient formation of short helix-like segments, and identify unique sequence segments important for protein folding. PMID- 17910958 TI - Role of Ser216 in the mechanism of action of membrane-bound lytic transglycosylase B: further evidence for substrate-assisted catalysis. AB - Lytic transglycosylases cleave the beta-(1-->4)-glycosidic bond in the bacterial cell wall heteropolymer peptidoglycan between the N-acetylmuramic acid (MurNAc) and N-acetylglucosamine (GlcNAc) residues with the concomitant formation of a 1,6 anhydromuramoyl residue. Based on sequence alignments, Ser216 in Pseudomonas aeruginosa membrane-bound lytic transglycosylase B (MltB) was targeted for replacement with alanine to delineate its role in the enzyme's mechanism of action. The specific activity of the Ser216-->Ala MltB derivative was less than 12% of that for the wild-type enzyme, while its substrate binding affinity remained virtually unaltered. These data are in agreement with a role of Ser216 in orienting the N-acetyl group on MurNAc at the -1 subsite of MltB for its participation in a substrate-assisted mechanism of action. PMID- 17910957 TI - Interaction of a fragment of the cannabinoid CB1 receptor C-terminus with arrestin-2. AB - Desensitization of the cannabinoid CB1 receptor is mediated by the interaction with arrestin. In this study, we report the structural changes of a synthetic diphosphorylated peptide corresponding to residues 419-439 of the CB1 C-terminus upon binding to arrestin-2. This segment is pivotal to the desensitization of CB1. Using high-resolution proton NMR, we observe two helical segments in the bound peptide that are separated by the presence a glycine residue. The binding we observe is with a diphoshorylated peptide, whereas a previous study reported binding of a highly phosphorylated rhodopsin fragment to visual arrestin. The arrestin bound conformations of the peptides are compared. PMID- 17910959 TI - Oleic acid: an efficient inhibitor of glucosyltransferase. AB - Among the extracts from 420 kinds of herbs, Prunus salicina, showing the highest glucosyltransferase inhibition activity, was purified and designated GTI-0163. Structural determination of GTI-0163 revealed it to be an oleic acid-based unsaturated fatty acid. GTI-0163 was an uncompetitive inhibitor of GTase. Among the unsaturated fatty acids, oleic acid showed a significantly higher GTase inhibitory activity than the saturated fatty acids or the ester form of oleic acid. These results strongly suggested that both the number of double bonds and the existence of free carboxyl groups of fatty acids play an important role in GTase inhibitory activity. PMID- 17910960 TI - Location of PsbY in oxygen-evolving photosystem II revealed by mutagenesis and X ray crystallography. AB - PsbY is one of the low molecular mass subunits of oxygen-evolving photosystem II (PSII). Its location, however, has not been identified in the current crystal structure of PSII. We constructed a PsbY-deletion mutant of Thermosynechococcus elongatus, crystallized, and analyzed the crystal structure of the mutant PSII dimer. The results obtained showed that PsbY is located in the periphery of PSII close to the alpha- and beta-subunits of cytochrome b559, which corresponded to an unassigned helix in the 3.7A structure of T. vulcanus or helix X2 in the 3.0A structure of T. elongatus. Our results also indicated that the C-terminal loop of PsbY is protruded toward the stromal side, instead of the lumenal side predicted previously. PMID- 17910962 TI - Urinary adrenalin and cortisol secretion patterns of social voles in response to adrenergic blockade under basal conditions. AB - The effect of alpha(1)- and beta-adrenergic blockade on daily rhythms of urinary adrenalin (ADR) and cortisol (CORT) under basal conditions were evaluated. Voles acclimated to a 12:12 h light/dark cycle at 26+/-2 degrees C received a single dose of either propranolol (PROP; 4.5 mg/kg) or prazosin (PRAZ; 1 mg/kg) 1 h before lights off. Urine samples were collected for 24 h at 4 h intervals. PROP evokes a significant increase in mean urinary ADR; although CORT was unaffected by PROP, PRAZ administration significantly decreased both urinary ADR and CORT during the scotophase as compared with control voles. Cosinor analysis indicated a significant 24 h rhythm in urinary ADR, but not in CORT secretion. ADR mesor and amplitude were increased and acrophase was significantly delayed by 5 h in PROP-treated voles; PRAZ elicited opposite effects. Unexpectedly, these changes in the 24 h ADR rhythm persisted 4-weeks after PROP-treatment. The 24 h rhythm components of urinary CORT were marginally altered 4-weeks post-PROP, but only the acrophase showed a significant change. Collectively, the results indicate that sympathetic activity has a redundant compensatory mechanism defending against physiological changes induced by beta-blockade. The simultaneous decrease in adrenal hormones induced by PRAZ suggests that alpha(1)-adrenoceptors may contribute to the mechanism of integrated stress responses. PMID- 17910961 TI - Ligand-transporter interaction in the AcrB multidrug efflux pump determined by fluorescence polarization assay. AB - The AcrB of Escherichia coli pumps out a wide range of compounds, including most of the currently available antibiotics, and contributes significantly to the serious problem of multidrug resistance of pathogenic bacteria. Quantitative analysis of drug efflux by this pump requires the measurement of the affinity of ligands. Yet there has been no success in determining these values. We introduce here an approach of steady-state fluorescence polarization to study the interactions between four different ligands and the purified AcrB transporter in a detergent environment. Our assays indicate that the transporter binds these drugs with K(D) values ranging from 5.5 to 74.1microM. PMID- 17910963 TI - Role of Tyr residues on the protein surface of cationic cell-wall-peroxidase (CWPO-C) from poplar: potential oxidation sites for oxidative polymerization of lignin. AB - It was previously reported that an unique peroxidase isoenzyme, cationic cell wall-bound peroxidase (CWPO-C), from poplar callus oxidizes sinapyl alcohol, ferrocytochrome c and synthetic lignin polymers, unlike other plant peroxidases. Here, the catalytic mechanism of CWPO-C was investigated using chemical modification and homology modeling. The simulated CWPO-C structure predicts that the entrance to the heme pocket of CWPO-C is the same size as those of other plant peroxidases, suggesting that ferrocytochrome c and synthetic lignin polymers cannot interact with the heme of CWPO-C. Since Trp and Tyr residues are redox-active, such residues located on the protein surface were predicted to be active sites for CWPO-C. Modification of CWPO-C Trp residues did not suppress its oxidation activities toward guaiacol and syringaldazine. On the other hand, modification of CWPO-C Tyr residues using tetranitromethane strongly suppressed its oxidation activities toward syringaldazine and 2,6-dimethoxyphenol by 90%, respectively, and also suppressed its guaiacol oxidation activity to a lesser extent. Ferrocytochrome c was not oxidized by Tyr-modified CWPO-C. These results indicate that the Tyr residues in CWPO-C mediate its oxidation of syringyl compounds and high-molecular-weight substrates. Homology modeling indicates that Tyr-177 and Tyr-74 are located near the heme and exposed on the protein surface of CWPO-C. These results suggest that Tyr residues on the protein surface are considered to be important for the oxidation activities of CWPO-C with a wide range of substrates, and potentially unique oxidation sites for the plant peroxidase family. PMID- 17910964 TI - Transport of camptothecin in hairy roots of Ophiorrhiza pumila. AB - We have investigated the subcellular accumulation and transport of camptothecin (CPT), a monoterpene indole alkaloid, in hairy roots of Ophiorrhiza pumila. This hairy root produces high amounts of CPT and excretes it into the culture medium. When the hairy roots were exposed to UV radiation, autofluorescence emitted from CPT showed subcellular localization of CPT in the vacuole. Treatment with several inhibitors suggested that CPT excretion is a transporter-independent passive transport controlled by the concentration gradient of the compound. Interestingly, the hairy roots treated with brefeldin A, a vesicle transport inhibitor, showed increased CPT excretion. This could be explained by an increased transport rate of CPT from the endoplasmic reticulum (ER) to the cytoplasm when transport of CPT to the vacuole is blocked. The much higher concentration of CPT in the cytoplasm resulted in the increased excretion rate. This result indicates that CPT is biosynthesized at the ER and transported to accumulate in the vacuole by the same machinery that is used for vacuolar protein sorting. How O. pumila is insensitive to CPT is discussed. PMID- 17910965 TI - Characterization of short-chain poly3-hydroxybutyrate in baker's yeast. AB - A short-chain poly3-hydroxybutyrate including four comonomers, originating from a complex with calcium polyphosphate, was isolated from commercial baker's yeast cells (Saccharomyces cerevisiae) and characterized as the second complexed poly(3 hydroxyalkanoate) (cPHA) in eukaryotes. The number-average molecular weight of 4982.5 Da with a polydispersity index of 1.11 was much lower than that of beet cPHA previously isolated. End-group analysis suggested that at least 60% of the molecules form the cyclic structures. Here, the organism-dependent structural diversity of cPHAs was completely established. It was also found that a change of culture medium influences the molecular weight but not the polydispersity of baker's yeast cPHA. PMID- 17910966 TI - Pharmacokinetics of disodium-fosfomycin in mongrel dogs. AB - Pharmacokinetic variables of fosfomycin were determined after administration of buffered disodium-fosfomycin intravenously (IV), intramuscularly (IM), subcutaneously (SC) and orally (PO), in mongrel dogs, at 40 and 80 mg/kgday for three days. Renal integrity was also assessed by measuring key serum variables. Day 1, day 2 and day 3 plasma concentration vs. time profiles were undistinguishable, but there appears to be a lineal increase in serum concentrations vs. time with the dose. A non-accumulative kinetic behavior was observed after three days with both doses and most pharmacokinetic variables remain unaltered. Considering a MIC range from 1 mirog/mL to 16 microg/mL of fosfomycin in serum for sensitive bacteria, and a negligible plasma protein binding of fosfomycin (<0.5%), useful plasma concentrations can only be achieved after the SC injection of 80 mg/kg every 12h, having a C(max)=18.96+/-0.3 microg/mL; a T(1/2beta)=2.09+/-0.06 microg/mL and a bioavailability of 84-85%. No alterations were observed in serum variables of kidney-related biochemical values. PMID- 17910967 TI - Canine and feline trefoil factor family peptides: highly conserved molecules with some unique characteristics. AB - Trefoil factor family (TFF) peptides maintain and repair the gastrointestinal mucosa and are aberrantly expressed in human and rodent inflammatory bowel disease and carcinomas, diseases common in dogs and cats. Study objectives were to sequence and translate canine and feline tff cDNAs and define any unique residues that might influence their structure and/or function. After isolation and reverse transcription of canine and feline gastrointestinal mucosal RNA, TFF cDNAs were amplified, sequenced, and cloned. Dogs and cats had unique amino acids in several places that were highly or completely conserved in other mammals, including a hydrophobic area in the TFF1 functional site, loop 2 of each TFF2 trefoil domain, a TFF3 dimerization site, and the TFF2 C-terminus. By identifying conserved and unique characteristics of canine and feline TFFs, this study establishes a foundation for investigation of dog and cat models of TFF-related diseases in both human and veterinary medicine. PMID- 17910968 TI - Cushing's disease in dogs: cabergoline treatment. AB - The treatment of pituitary-dependent hyperadrenocorticism (PDH) in dogs has for a long time been focused on inhibiting the adrenal gland using drugs such as o-p' DDD, Ketoconazole and Trilostane, without attacking the primary cause: the corticotrophinoma. Corticotroph cells can express the D2 dopaminergic receptor; therefore cabergoline (Cbg) could be effective as a treatment. Follow-up over 4 years was carried out in 40 dogs with PDH that were treated with Cbg (0.07 mg/kg/week. Out of the 40 dogs, 17 responded to Cbg (42.5%). A year after the treatment, there was a significant decrease in ACTH (p<0.0001), alpha-MSH (p<0.01), urinary cortisol/creatinine ratio (p<0.001), and of the tumor size (p<0.0001) evaluated by nuclear magnetic resonance. Dogs responding to Cbg lived significantly longer (p<0.001) than those in the control group. To conclude, Cbg is useful in 42.5% of dogs with PDH, justifying its use as a treatment. PMID- 17910970 TI - Effective elution of RDX and TNT from particles of Comp B in surface soil. AB - During live fire training exercises, large amounts of explosives are consumed. Low order detonations of high explosive payloads result in the patchy dispersal of particles of high explosive formulations over large areas of firing range soils. Dissolution of explosives from explosive formulation particles into soil pore water is a controlling factor for transport, fate, and effects of explosive compounds. We developed an empirical method to evaluate soils based on functionally defined effective dissolution rates. An automated Accelerated Solvent Extractor was used to determine the effective elution rates under controlled conditions of RDX and TNT from soil columns containing particles of Comp B. Contrived soils containing selected soil geosorbants and reactive surfaces were used to quantitatively determine the importance of these materials. Natural soils from training ranges of various soil types were also evaluated. The effects of geosorbants on effective elution rates were compound- and sorbent specific. TNT elution was less than that of RDX and was greatly slowed by humic acid. Iron and iron-bearing clays reduced the effective elution rates of both RDX and TNT. This empirical method is a useful tool for directly generating data on the potential for explosives to leach from firing range soils, to identify general bulk soil characteristics that can be used to predict the potential, and to identify means to engineer soil treatments to mitigate potential transport. PMID- 17910969 TI - Modeling photothermal and acoustical induced microbubble generation and growth. AB - Previous experimental studies showed that powerful heating of nanoparticles by a laser pulse using energy density greater than 100 mJ/cm(2), could induce vaporization and generate microbubbles. When ultrasound is introduced at the same time as the laser pulse, much less laser power is required. For therapeutic applications, generation of microbubbles on demand at target locations, e.g. cells or bacteria can be used to induce hyperthermia or to facilitate drug delivery. The objective of this work is to develop a method capable of predicting photothermal and acoustic parameters in terms of laser power and acoustic pressure amplitude that are needed to produce stable microbubbles; and investigate the influence of bubble coalescence on the thresholds when the microbubbles are generated around nanoparticles that appear in clusters. We develop and solve here a combined problem of momentum, heat and mass transfer which is associated with generation and growth of a microbubble, filled with a mixture of non-vaporized gas (air) and water vapor. The microbubble's size and gas content vary as a result of three mechanisms: gas expansion or compression, evaporation or condensation on the bubble boundary, and diffusion of dissolved air in the surrounding water. The simulations predict that when ultrasound is applied relatively low threshold values of laser and ultrasound power are required to obtain a stable microbubble from a single nanoparticle. Even lower power is required when microbubbles are formed by coalescence around a cluster of 10 nanoparticles. Laser pulse energy density of 21 mJ/cm(2) is predicted for instance together with acoustic pressure of 0.1 MPa for a cluster of 10 or 62 mJ/cm(2) for a single nanoparticle. Those values are well within the safety limits, and as such are most appealing for targeted therapeutic purposes. PMID- 17910971 TI - Spatial variation of DDT and its metabolites in fish and sediment from Cinca River, a tributary of Ebro River (Spain). AB - The Cinca River is a tributary of Ebro River in the NE of Spain which receives input from different activities. One of the most important is related to an industry where DDT is used as an intermediate in the production of dicofol. This study evaluated the DDT contamination along the Cinca River. Sampling sites were selected up- and downstream from this industry. Sediments and fishes (59 bleaks (Alburnus alburnus) and 23 barbels (Barbus graellsi)) were collected in 2002 and analyzed using a new and rapid selective pressurized liquid extraction (SPLE) method. DDT and its metabolites were found in sediments and fishes at levels ranging from 9 to 94microg kg(-1) dry weight (d.w.) and from not detected to 2098microg kg(-1) wet weight (w.w.), respectively. The highest values corresponded to samples collected just downstream the industry. Thirty kilometers downstream from the factory, levels were clearly lower, showing a weakening of the impact. p,p'-DDE isomer comprised up to 50% and 70% of total DDT measured in sediment and fish, respectively. When compared with values obtained in a previous study in 1999, a generalized drop of the levels in all matrixes (77-97%), was observed. No meaningful differences were found between the two fish species studied neither between the two tissues (muscle and liver) analyzed. PMID- 17910972 TI - Well water contaminated by acidic mine water from the Dabaoshan Mine, south China: chemistry and toxicity. AB - An investigation into well water quality was carried out in a rural area subject to irrigation with acidic mine water from the Guangdong Dabaoshan Mine, southern China. The results of water pH measurements from 112 wells in two different seasons suggest that the well water has been contaminated to varying degrees in the investigated Shangba floodplain (approximately 11km south of the Guangdong Dabaoshan Mine). There is a trend that well water pH increased southwards, suggesting that the impacts of acidic irrigation water on groundwater decreased with increasing distance to the entry point of acidic irrigation water. Water quality monitoring results of the selected wells show that Cu and Cd in the water exceeded the limits set in the Chinese National Standards for Drinking Water (GB 5749-85) for the wells close to the irrigation water source. If the World Health Organization (WHO) standard was considered, Cd in some wells was almost 10 times as high as the WHO guideline value (0.003mg l(-1)). Water collected from the location closest to the acidic irrigation water source was acutely toxic to the test organism (Daphnia carinata) even after 51 time dilution. It is likely that the extremely high mortality rate of the local population reported for the study area is at least partly related to the high levels of heavy metals, particularly Cd in the drinking well water. PMID- 17910973 TI - Occurrence of polychlorinated naphthalenes, polychlorinated biphenyls and short chain chlorinated paraffins in marine sediments from Barcelona (Spain). AB - Polychlorinated naphthalenes (PCNs), short-chain chlorinated paraffins (SCCPs) and polychlorinated biphenyls (PCBs) were analysed in marine sediment samples collected from the coastal area of Barcelona (Spain) and near of a submarine emissary coming from a waste water treatment plant located at the mouth of the Besos River (Barcelona). An integrated sample treatment based on Soxhlet extraction followed by a simple clean-up with Florisil and graphitized carbon cartridge was employed. Gas chromatography coupled to ion-trap tandem mass spectrometry (GC-MS/MS) and gas chromatography-mass spectrometry in electron capture negative ionization mode, were used for PCN and SCCP determinations, respectively, while for PCB analysis gas chromatography with electron capture detection (GC-ECD) was used. The method developed provided low limits of detection (0.001-0.003 ng g(-1) dry weight (dw) for PCNs, 1.8 ng g(-1) for SCCPs and 0.006-0.014 ng g(-1)dw for PCBs) and good run-to-run precisions (lower than RSD 8%) for the analysis of sediment samples. Concentration levels ranging from 0.17 to 3.27 ng g(-1)dw for PCNs, between 0.21 and 1.17 microg g(-1)dw for SCCPs, and from 2.33 to 44.00 ng g(-1) (dw) for PCBs, were found in the coastal sediments, while for samples collected near to the submarine emissary higher levels (from 2.02 to 6.56 ng g(-1)dw for PCNs, between 1.25 and 2.09 microg g( 1)dw for SCCPs and from 22.34 to 37.74 ng g(-1)dw for PCBs) were obtained. The results obtained provide new data about the occurrence of PCN and SCCP in the coastal area of Barcelona. PMID- 17910974 TI - The physico-chemical properties and leaching behaviors of phosphatic clay for immobilizing heavy metals. AB - In this study, phosphatic clay was used as a phosphate containing material. The fractionation of phosphorus was carried out using the CRM BCR-684 protocol, and the inorganic phosphorus, especially all the apatite phosphorus, was found as the major form. The elemental compositions of the phosphatic clay were identified using an X-ray fluorescence spectrometer, and was found to be mainly composed of CaO and P2O5. The specific surface area, pore volume and average pore diameter were measured also. Results of experiment show that the phosphatic clay may provide a cost-effective way to remediate heavy metal contaminated aqueous and slurry phase. PMID- 17910975 TI - Development and application of kinetic model on biological anoxic/aerobic filter. AB - An up-flow biological anoxic filter (BANF) has been developed to achieve high removal performance of suspended solids and BOD removal as well as nitrogen. With a view to understand treatment mechanisms, we developed a filtration model that incorporates filtration, deposit scoring and biological reactions simultaneously. The biological reactions consist of four types of reaction; dissolution of organic particles; utilization of dissolved organic matter; denitrification; and self-degradation of bacteria. Whereas the reactor is generally assumed to be a plug flow reactor in the filtration model, it is assumed a continuous-flow stirred tank reactor (CSTR) in the model of biological reactions. The hydrodynamics is supposed that the filter bottom (the portion sludge settled) is a CSTR and the filter bed (the portion filled with filter media) consists of number of CSTR of equal size arranged in series. The model obtained in this study was verified and simulated using experimental results taken from a pilot-scale plant and predicted the experimental data well, applying to design and operate BANF. PMID- 17910976 TI - Titanium dioxide mediated photocatalytic degradation of monochlorobenzene in aqueous phase. AB - The reaction sequence for the photocatalytic degradation of monochlorobenzene (MCB) in UV/TiO2 process, including substrate adsorption, degradation, and mineralization, was studied. The theoretical maximum quantity of MCB that could be adsorbed onto TiO2 surface in aqueous phase was 0.18+/-0.04 micromol m(-2) of TiO2. In accordance with the upper limit of the relative surface coverage of MCB molecules to surface hydroxyls of TiO2 was around 2.2%, the water molecules as the major adjacent species near TiO2 surface would compete with MCB molecules. Increasing the initial substrate concentration to an appropriate value or enhancing the affinity between the MCB and the TiO2 surface by adjusting the solution pH would promote the photocatalytic degradation. Experimental results revealed that the neutral medium was beneficial for the degradation of MCB. In comparison, the mineralization was most improved at acidic condition. Generally, 90% of the total organic carbon (TOC) was mineralized after 240 min illumination time in the examined pH range except solution pH 11. The suppressed mineralization of MCB at solution pH 11 was ascribed to the lack of adsorption. A simplified 2-step consecutive kinetic model was used to simulate the mineralization. PMID- 17910977 TI - Comparison between laboratory and field leachability of MSWI bottom ash as a road material. AB - The leaching properties of bottom ash from municipal solid waste incineration (MSWI) used as an aggregate substitute in unbound pavement layers are evaluated. The mechanical behaviour of bottom ash is acceptable for this application, but the potential environmental consequences constitute the most important limitation on the use of bottom ash as a road material. The environmental properties of bottom ash are assessed by means of the Dutch availability test NEN 7341 and the single-batch and two-stage batch European EN 12457 laboratory leaching tests. Furthermore, an experimental unbound pavement stretch is constructed to provide information on leaching behaviour under field conditions. In this high infiltration scenario, the results from predicted (based upon laboratory leaching tests) and measured releases (under field conditions) are compared, evidencing that predictions based on compliance leaching tests may be highly realistic. The depletion period of the extractable fraction of a number of elements in these field conditions is also quantified. PMID- 17910978 TI - Chemical composition of enamel and dentine in primary teeth in children from Thailand exposed to lead. AB - Enamel and dentine in teeth of children with high blood levels of lead were analyzed by means of secondary ion mass spectrometry (SIMS) and X-ray micro analyses (XRMA) and compare with teeth from children with low blood levels of lead. The SIMS analysis revealed detectable levels of Pb in dentine close to the pulp. The XRMA analyses could not detect any lead. There were no differences found in lead level in enamel of high lead level exposed teeth from low level exposed. The results confirm that children with high blood levels of lead have an uptake of lead in dentine close to the pulp. PMID- 17910979 TI - Assessment and management of risk to wildlife from cadmium. AB - Cadmium, a nonessential heavy metal that comes from natural and anthropogenic sources, is a teratogen, carcinogen, and a possible mutagen. Assessment of potential risk from cadmium requires understanding environmental exposure, mainly from ingestion, although there is some local exposure through inhalation. Chronic exposure is more problematic than acute exposure for wildlife. There is evidence for bioaccumulation, particularly in freshwater organisms, but evidence for biomagnification up the food chain is inconsistent; in some bird studies, cadmium levels were higher in species that are higher on the food chain than those that are lower. Some freshwater and marine invertebrates are more adversely affected by cadmium exposure than are birds and mammals. There is very little experimental laboratory research on the effects of cadmium in amphibians, birds and reptiles, and almost no data from studies of wildlife in nature. Managing the risk from cadmium to wildlife involves assessment (including ecological risk assessment), biomonitoring, setting benchmarks of effects, regulations and enforcement, and source reduction. PMID- 17910980 TI - Effects of bortezomib on platelet aggregation and ATP release in human platelets, in vitro. AB - INTRODUCTION: Proteasome inhibitor bortezomib (PS-341) has been the first proteasome inhibitor that has entered clinical trials with its antiproliferative and proapoptotic effects in patients with multiple myeloma. Recent studies indicate that proteasome inhibitors can be useful in prevention of experimental arterial thrombosis in renovascular hypertensive rat models. The aim of the present study is to investigate the effect of bortezomib on in vitro platelet aggregation and adenosine triphosphate (ATP) release of human platelets. MATERIALS AND METHODS: For this purpose, platelet aggregation was induced in the platelet-rich plasma (PRP) using 3 microg ml(-1) collagen, 5 microM adenosine diphosphate (ADP), 10 microM epinephrine and 1 U ml(-1) thrombin and ATP release was induced by collagen. RESULTS AND CONCLUSIONS: Bortezomib showed an inhibitory effect on platelet aggregation induced by ADP in human PRP in a dose- and time dependent manner, whereas it had no effect on collagen-, epinephrin and thrombin induced aggregation. ATP-release reaction induced by collagen was inhibited dose- and time-dependently by bortezomib, even though collagen-induced platelet aggregation was apparently not affected in human PRP. These findings indicate that bortezomib may be an antiaggregating agent and its' effects may be related to adenine nucleotide receptor dependent regulatory proteins which are important for physiological and pathophysiological cellular processes. However, our in vitro studies suggest that this hypothesis is inadequate to explain the observations completely. This phenomenon and its clinical implication justify further clinical investigations. PMID- 17910981 TI - Combination therapy with pegylated liposomal doxorubicin and carboplatin in gynecologic malignancies: a prospective phase II study of the Arbeitsgemeinschaft Gynaekologische Onkologie Studiengruppe Ovarialkarzinom (AGO-OVAR) and Kommission Uterus (AGO-K-Ut). AB - OBJECTIVE: A multicenter non-randomized phase II study was initiated to evaluate tolerability and efficacy of pegylated liposomal doxorubicin (PLD) in combination with carboplatin in gynecologic malignancies. METHODS: One hundred forty women with recurrent or advanced endometrial (n=31), cervical or vaginal cancer (n=31), uterine sarcomas (n=11), or recurrent platinum-sensitive ovarian cancer (n=67) received six courses of PLD 40 mg/m2 and carboplatin (AUC 6) every 28 days. RESULTS: Hematological toxicities with NCI-CTC grade 3/4 were anemia in 8%, thrombocytopenia in 14%, neutropenia in 24%, and febrile neutropenia in 2% of 652 cycles. Grade 3/4 non-hematological toxicities included fatigue (14% of patients), pain (10%), dyspnea (9%), palmar-plantar erythrodysesthesia (7%), and nausea/vomiting (7%). Dose intensity reached 87.2% for PLD and 88.2% for carboplatin. Seventy-four percent of all non-progressive patients received at least 5 cycles. Overall response rates were (116 patients evaluable for response): ovarian cancer (n=54) 68%, endometrial cancer (n=27) 44%, uterine sarcomas (n=9) 33%, and cervical/vaginal cancer (n=26) 12%. Median progression free survival was 11.6 months (95%CI 9.6-14.1) for ovarian cancer and 9.5 months (95%CI 6.6-12.6) for endometrial cancer. Median overall survival was 23.8 months (95%CI 19.0-30.2) and 21.4 months (95%CI 11.9-), respectively. CONCLUSIONS: The combination of PLD and carboplatin was well tolerated and feasible in patients with gynecologic malignancies. Efficacy was low in cervical/vaginal cancer, but promising in patients with endometrial cancer. Efficacy was within the expected range in recurrent platinum-sensitive ovarian cancer and is currently under further investigation in a prospective randomized phase III trial comparing PLD/carboplatin with paclitaxel/carboplatin (CALYPSO-trial; AGO-OVAR 2.9). PMID- 17910982 TI - Orexin neuronal circuitry: role in the regulation of sleep and wakefulness. AB - Orexin A and orexin B were initially identified as endogenous ligands for two orphan G protein-coupled receptors [104]. They were initially recognized as regulators of feeding behavior in view of their exclusive production in the lateral hypothalamic area (LHA), a region known as the feeding center, and their pharmacological activity [104,30,49,107]. Subsequently, the finding that orexin deficiency causes narcolepsy in humans and animals suggested that these hypothalamic neuropeptides play a critical role in regulating sleep/wake cycle [22,46,71,95,117]. These peptides activate waking-active monoaminergic and cholinergic neurons in the hypothalamus/brain stem regions to maintain a long, consolidated awake period. Recent studies on efferent and afferent systems of orexin neurons, and phenotypic characterization of genetically modified mice in the orexin system further suggested roles of orexin in the coordination of emotion, energy homeostasis, reward system, and arousal [3,80,106,137]. A link between the limbic system and orexin neurons might be important for increasing vigilance during emotional stimuli. Orexin neurons are also regulated by peripheral metabolic cues, including ghrelin, leptin, and glucose, suggesting that they might have important roles as a link between energy homeostasis and vigilance states [137]. Recent research has also implicated orexins in reward systems and the mechanisms of drug addiction [13,48,91]. These observations suggest that orexin neurons sense the outer and inner environment of the body, and maintain proper wakefulness of animals for survival. This review discusses the mechanism by which orexins maintain sleep/wakefulness states, and how this mechanism relates to other systems that regulate emotion, reward, and energy homeostasis. PMID- 17910983 TI - Unattended emotional intonations modulate linguistic prosody processing. AB - Prosody or speech melody subserves linguistic (e.g., question intonation) and emotional functions in speech communication. Findings from lesion studies and imaging experiments suggest that, depending on function or acoustic stimulus structure, prosodic speech components are differentially processed in the right and left hemispheres. This direct current (DC) potential study investigated the linguistic processing of digitally manipulated pitch contours of sentences that carried an emotional or neutral intonation. Discrimination of linguistic prosody was better for neutral stimuli as compared to happily as well as fearfully spoken sentences. Brain activation was increased during the processing of happy sentences as compared to neutral utterances. Neither neutral nor emotional stimuli evoked lateralized processing in the left or right hemisphere, indicating bilateral mechanisms of linguistic processing for pitch direction. Acoustic stimulus analysis suggested that prosodic components related to emotional intonation, such as pitch variability, interfered with linguistic processing of pitch course direction. PMID- 17910984 TI - Should targeting immunosuppression, immunoregulation or remyelination be used to treat inflammatory neuropathies? PMID- 17910985 TI - Walking on music. AB - The present study focuses on the intricate relationship between human body movement and music, in particular on how music may influence the way humans walk. In an experiment, participants were asked to synchronize their walking tempo with the tempo of musical and metronome stimuli. The walking tempo and walking speed were measured. The tempi of the stimuli varied between 50 and 190 beats per minute. The data revealed that people walk faster on music than on metronome stimuli and that walking on music can be modeled as a resonance phenomenon that is related to the perceptual resonance phenomenon as described by Van Noorden and Moelants (Van Noorden, L., & Moelants, D. (1999). Resonance in the perception of musical pulse. Journal of New Music Research, 28, 43-66). PMID- 17910986 TI - The effect of micro-architectural structure of cabbage substratum and or background bacterial flora on the growth of Listeria monocytogenes. AB - The effect of micro-architectural structure of cabbage (Brassica oleracea var. capitata L.) substratum and or background bacterial flora on the growth of Listeria monocytogenes as a function of incubation temperature was investigated. A cocktail mixture of Pseudomonas fluorescens, Pantoea agglomerans and Lactobacillus plantarum was constituted to a population density of approximately 5 log CFU/ml in order to pseudo-simulate background bacterial flora of fresh-cut cabbage. This mixture was co-inoculated with L. monocytogenes (approximately 3 log CFU/ml) on fresh-cut cabbage or in autoclaved cabbage juice followed by incubation at different temperatures (4-30 degrees C). Data on growth of L. monocytogenes were fitted to the primary growth model of Baranyi in order to generate the growth kinetic parameters of the pathogen. During storage, microbial ecology was dominated by P. fluorescens and L. plantarum at refrigeration and abuse temperature, respectively. At all temperatures investigated, lag duration (lambda, h), maximum specific growth rate (micro(max), h(-1)) and maximum population density (MPD, log CFU/ml) of L. monocytogenes were only affected by medium micro-architectural structure, except at 4 degrees C where it had no effect on the micro(max) of the pathogen. Comparison of observed values of micro(max) with those obtained from the Pathogen Modelling Program (PMP), showed that PMP overestimated the growth rate of L. monocytogenes on fresh-cut cabbage and in cabbage juice, respectively. Temperature dependency of micro(max) of L. monocytogenes, according to the models of Ratkowsky and Arrhenius, showed linearity for temperature range of 4-15 degrees C, discontinuities and linearity again for temperature range of 20-30 degrees C. The results of this experiment have shown that the constituted background bacterial flora had no effect on the growth of L. monocytogenes and that micro-architectural structure of the vegetable was the primary factor that limited the applicability of PMP model for predicting the growth of L. monocytogenes on fresh-cut cabbage. A major limitation of this study however is that nutrient profile of the autoclaved cabbage juice may be different from that of the raw juice thus compromising realistic comparison of the behaviour of L. monocytogenes as affected by micro architectural structure. PMID- 17910987 TI - Effects of collagen crosslinking on tissue fragility. PMID- 17910988 TI - Spinal unloading after abdominal exercises. AB - BACKGROUND: Intervertebral discs are exposed to compressive forces, which produces fluid loss. This loss decreases disc height, spinal length and consequently overall stature. The loss of stature has been associated with spine loading and low back pain. Abdominal exercises increase intra-abdominal pressure and unload the spine. The purpose of this study was to identify if abdominal exercises may produce acute spinal unloading compared to a known unloading position. METHODS: Nine subjects performed a loading protocol and an unloading protocol on three experimental sessions. The loading protocol consisted of three sets of military press, while three unloading protocols were: three sets of regular abdominal exercises, three sets of abdominal exercises performed in an inclined board or an unloading resting posture. FINDINGS: Abdominal exercises showed a greater recovery (mean (SD)) (regular=87.8 (20.4)%; inclined=70.1 (14.5)%) in comparison to the resting position (Fowler=33.6 (14.1)%), although no significant differences were found between abdominal exercises in stature recovery (P=0.07). INTERPRETATION: Abdominal exercises may be introduced between sets of resistance training to decrease the negative effect of compressive forces imposed during such highly stressing activities. Further studies are required to confirm the proposed underlying mechanisms. PMID- 17910989 TI - Comparative genotoxicity evaluation of imidazolinone herbicides in somatic cells of Drosophila melanogaster. AB - In the present study, five analogous herbicides, namely Imazapyr (IMZR), Imazapic (IMZC), Imazethapyr (IMZT), Imazamox (IMZX) and Imazaquin (IMZQ), were evaluated for genotoxicity (mutagenic and recombinagenic activity) in the wing somatic mutation and recombination test (SMART) of Drosophila melanogaster. They are classified as imidazolinone (IMI) herbicides and their mode of action is to inhibit acetohydroxyacid synthase (AHAS), an enzyme involved in the biosynthesis of the amino acids leucine, isoleucine and valine. Two crosses were used: the standard (ST) cross and the high bioactivation (HB) cross. The latter is characterized by high levels of cytochrome P450 conferring increased sensitivity to promutagens and procarcinogens. Three-day-old larvae were exposed by chronic feeding (48 h) to four different concentrations of these herbicides (2.5, 5.0, 10.0 or 20.0 mM). For the evaluation of genotoxic effects, the frequencies of spots per individual in the treated series were compared to the concurrent negative control series (ultrapure water). Imazapyr, Imazapic and Imazethapyr gave negative results with both crosses of the wing spot test. In the ST cross, Imazamox showed positive results only for large single spots (20.0 mM IMZX) and weak positive results for total spots (10.0 and 20.0 mM IMZX), while Imazaquin showed positive results only for large single spots (5.0 and 20.0mM IMZQ) and a weak positive result for total spots (20.0 mM IMZQ). These positive results are mainly due to induced recombination and to a minor extent to mutations. In the HB cross, only Imazamox (5.0 mM IMZX) showed a weak positive result for small single spots. The positive control urethane, a promutagen, caused an increase in the number of all types of spots in both crosses. In conclusion, the results of chronic treatments performed at high doses (toxicity was observed at higher doses) shows the existence of a genotoxic risk for IMZX and IMZQ exposure under these experimental conditions, and indicate the need for further research to delineate the exact mechanisms involved. PMID- 17910990 TI - Hypoxia induces upregulation of the deoxyribonuclease I gene in the human pancreatic cancer cell line QGP-1. AB - We have previously demonstrated that ischemia caused by acute myocardial infarction induces an abrupt increase of serum deoxyribonuclease I (DNase I) activity. In this study, we examined whether hypoxia can affect the levels of DNase I activity and/or its transcripts in vitro. We first exposed the human pancreatic cancer cell line QGP-1, which is the first documented DNase-I producing cell line, to hypoxia (2% O2), and found that this induced a significant increase in both the activity and transcripts of DNase I. This response was mediated by increased transcription only from exon 1a of the two alternative transcription-initiating exons utilized simultaneously in the human DNase I gene (DNASE1); exposure of QGP-1 cells to hypoxia for 24 h resulted in a 15-fold increase of DNASE1 transcripts starting from exon 1a compared with the expression level under normoxic conditions. Promoter, electrophoretic mobility shift, and chromatin immunoprecipitation assays with QGP-1 cells exposed to hypoxia or normoxia showed that the region just upstream from exon 1a was involved in this response in a hypoxia-induced factor-1-independent, but at least in a Sp1 transcription factor-dependent manner possibly through enhanced binding of Sp1 protein to the promoter. These results indicate that DNASE1 expression is upregulated by hypoxia in the cells. PMID- 17910991 TI - A hypothesis: adiponectin mediates anti-atherosclerosis via adventitia-AMPK-iNOS pathway. AB - Adiponectin is an adipocyte-derived protein with insulin-sensitizing, anti inflammatory, and anti-atherogenic properties and is abundantly found in plasma. Vascular adventitia is the outermost connective and supporting tissue of vessels. Recently, increasing evidence has shown that infection in the adventitia is one of the causes of atherosclerosis and restenosis. Our previous study indicated that local transferring adenovirus expressing adiponectin gene (Ad-APN) to intima and adventitia can suppress atherosclerosis, but the exact mechanism is still obscure. We speculate that with infection in the adventitia, adiponectin can activate AMP-activated protein kinase (AMPK) through adiponectin receptors in the membranes of adventitial fibroblasts and then inhibit the expression and activity of inducible nitric oxide synthase (iNOS); secretion of adventitial infective factors; division, proliferation and translation of adventitial fibroblasts; and change of adventitial fibroblasts to myofibroblasts, finally decreasing oxidative/nitrative stress to reduce atherosclerotic plaque area and stabilize atherosclerotic plaques. The proposition may provide clues into the development of a novel treatment for atherosclerosis. PMID- 17910992 TI - Truly personalised medicine: self-experimentation in medical discovery. AB - Biomedical research need not be carried out solely by 'Them': distant, dissociated, enormously costly institutions and companies. It can, and increasingly in the 21st century will, be carried out by 'Us', the informed non professional. Conventional clinical trials treat humans with the same experimental model as laboratory rats - regarding them as mute, variable, unreliable material from which results must be obtained as fast as possible to maximise return on investment and patent life. The alternative is longer term, self-reported clinical studies of new treatments, based on the assumption that the experimenter is informed, intelligent and aware. A wide variety of new treatments for chronic disease are available, involving elements of diet, behaviour, environment and non-prescription medication as well as ethical pharmaceuticals, and previous experience suggests that they can be enormously effective. The key is objective, quantifiable measures of outcome. These can be achieved with over-the-counter diagnostics for a variety of parameters, as well as with self-built test systems, and careful and systematic observations of symptoms. Hypothesis generation is a key part of this process. PMID- 17910993 TI - Mechanical stress and formation of protein aggregates in neurodegenerative disorders. AB - Misfolded protein aggregates and inclusion bodies have been associated with various protein conformation disorders such as Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and prion diseases including bovine spongiform encephalopathy (BSE). Models have been proposed as plausible explanations for the extension and progression of protein aggregates; however, little is known about the initiation process of protein aggregation, particularly in sporadic neurodegenerative diseases. Epidemiological data have suggested a tight association between sporadic neurodegenerative diseases and history of mechanical stresses such as trauma, head injury, and occupational exposures, including professional soccer and boxer's brain that carries histological hallmarks of AD/PD. Here, we propose that mechanical stress is an environmental factor that provokes a disturbance in cellular quality control systems and molecular chaperones that target misfolded proteins. This subsequently initiates protein aggregation and results in sporadic neurodegenerative disorders. Further, continuous and repetitive exposure to environmental mechanical stress, mostly in an unrecognized manner, is inevitable in daily life and thus, it functions as a potential driving force for protein aggregation. In this regard, a recent identification of the fact that an intracellular mechanosensor actually exists may support our notion. Reduction in the mechanical stress in combination with other conventional aspects should facilitate the development of rational therapeutics for these neurodegenerative disorders. PMID- 17910994 TI - Revisiting the Cartesian model of pain. AB - In modern medicine, the Cartesian or nociceptive concept of chronic pain has been replaced with the biopsychosocial model in both theory and practice. This paper presents an argument along with observations in favor of chronic pain as a pure nociceptive experience separate from suffering and outlines theoretical and practical solutions to improve the diagnosis and treatment of patients who experience chronic pain. Theoretical solutions include increasing inhibitory descending neurotransmitters using monoamine oxidase inhibitors of subtype A in combination with dextroamphetamine, increasing beta endorphin through enzymology and/or ultrasound stimulation of the periaqueductal gray, developing long duration opioid analgesics using spin label probes of morphine and morphine analogs and destructive interference of nociceptive action potentials by eddy currents generated by a variable magnetic field. Practical solutions include prolonging local anesthetic blockade of small pain fibers with patient administered local anesthetic storage devices and abandonment of the multidisciplinary pain clinic. PMID- 17910995 TI - A postulated role for human papillomavirus (HPV) in the transformation and proliferation of oral squamous cell carcinoma (OSCC). AB - Oral squamous cell carcinoma (OSCC) is the sixth most common malignancy and is a major cause of cancer morbidity and mortality worldwide. A strong association of oral cancer with high-risk HPVs infection underlines the importance of the virus in the pathogenesis of these squamous cell carcinomas. We postulate that HPV may contribute to the carcinogenesis of oral epithelial and function to stimulate transformation and proliferation of OSCC. The appropriate method, which may involve immunoprevention, immunotherapy, or immunomodulatory, needs to be developed. Not only humoral immunity, but also cellular immunity must be part of this process. PMID- 17910996 TI - Effect of subject task on contralateral suppression of click evoked otoacoustic emissions. AB - Contralateral suppression of click evoked otoacoustic emissions (CEOAEs) is widely used as a non-invasive measure of the activity of the (uncrossed) medial olivocochlear bundle (MOCB). There is evidence that the uMOCB receives descending input from the cortex, potentially mediating top-down control during higher order processing. This study investigated whether the contralateral suppression measure is affected by top-down influences during different tasks performed by the participants during recording. Suppression of CEOAEs evoked at 50 and 60dB SPL was measured under four different task conditions: (1) no task; (2) passive visual (watching a silent subtitled DVD); (3) active visual (responding to visually presented sums); (4) active auditory (detecting tone pips embedded in the evoking click train). The most significant effect of task was found on the recording noise, with both the passive visual and the active auditory task producing significantly lower noise levels than the no task condition. In the passive visual task, this was associated with a reduced inter-subject variability, which enhanced the effect size relative to the no task condition. A main effect of subject task was also found on the change in CEOAE I/O slope due to contralateral noise. This effect reflected a significantly smaller suppression during the active auditory task compared to the no task condition, leading to a reduced effect size. No significant difference in suppression strength between the no task condition and the two non-auditory tasks was observed, suggesting that the main effect of task reflects a specific effect of auditory attention. The data suggest that MOCB activity is inhibited due to top-down influences when selective attention is focussed on the ipsilateral ear. PMID- 17910998 TI - In vitro pharmacodynamics of moxifloxacin versus levofloxacin against 4 strains of Streptococcus pneumoniae: 1 wild type, 2 first-step parC mutants, and 1 pump mutant. AB - Levofloxacin binds topoisomerase IV, whereas moxifloxacin preferentially binds DNA gyrase. Most 1st-step pneumococcal mutants have alterations in the parC gene of topoisomerase IV. Because of differential binding affinity, moxifloxacin may have superior activity against 1st-step mutants compared with levofloxacin. The purpose of this work was to compare rates and extent of bacterial killing of genetically characterized Streptococcus pneumoniae with moxifloxacin and levofloxacin. Four strains of S. pneumoniae were used: a wild type, 2 first-step parC mutants, and a pump mutant. Using an in vitro pharmacodynamic model run in duplicate, we exposed bacteria to unbound moxifloxacin and levofloxacin peaks of 2 and 4.5 mg/L, respectively, which emulated clinical dosing. Additional experiments were done in which the area under the curve (AUC)/MIC ratio of 1 agent was matched to the competing drug's clinical dose AUC/MIC ratio. Time kill curves were analyzed for rate and extent of bacterial kill and regrowth. Pre- and postexposure MIC and polymerase chain reaction (PCR) testing were done. Moxifloxacin and levofloxacin displayed similar rates and extent of bacterial kill for the wild type, efflux pump type, and parC mutant 27-1361B. Moxifloxacin initially achieved a faster rate of kill, regardless of the AUC/MIC ratio, against parC mutant 7362 (P < 0.05) but not an advantage in time to 3 log kill. Postexposure MIC values were elevated for strain 7362 in 2 moxifloxacin experiments and 1 levofloxacin experiment. Post-PCR analysis revealed new gyrA mutations for all 3 isolates. Both moxifloxacin and levofloxacin are effective against multiple strains of S. pneumoniae. PMID- 17910997 TI - Cochlear implants and brain plasticity. AB - Cochlear implants have been implanted in over 110,000 deaf adults and children worldwide and provide these patients with important auditory cues necessary for auditory awareness and speech perception via electrical stimulation of the auditory nerve (AN). In 1942, Woolsey and Walzl presented the first report of cortical responses to localised electrical stimulation of different sectors of the AN in normal hearing cats, and established the cochleotopic organization of the projections to primary auditory cortex. Subsequently, individual cortical neurons in normal hearing animals have been shown to have well characterized input-output functions for electrical stimulation and decreasing response latencies with increasing stimulus strength. However, the central auditory system is not immutable, and has a remarkable capacity for plastic change, even into adulthood, as a result of changes in afferent input. This capacity for change is likely to contribute to the ongoing clinical improvements observed in speech perception for cochlear implant users. This review examines the evidence for changes of the response properties of neurons in, and consequently the functional organization of, the central auditory system produced by chronic, behaviourally relevant, electrical stimulation of the AN in profoundly deaf humans and animals. PMID- 17910999 TI - Rapid isothermal detection assay: a probe amplification method for the detection of nucleic acids. AB - Simple, accurate, and stable diagnostic tests are essential to control viral infectious diseases such as avian influenza virus. The current technologies are often inaccessible to people who need them, mainly because of the specialized equipment and the need for highly trained technologists. Here, we describe a rapid isothermal nucleic acid detection assay (RIDA) that can be used to detect both DNA and RNA targets. Using chemically modified probes, we designed a lateral flow (LF) immunoassay that can be used in combination with RIDA for equipment free nucleic acid target detection. RIDA is a "probe amplification" assay that uses the single-strand nicking activity of restriction nicking endonucleases to repeatedly cleave synthetic probes hybridizing to the same target sequences. In the RIDA-LF combined assay, chemically labeled probes are covalently conjugated to magnetic microbeads, which is propitious to separate cleaved probes from the reaction solution. The cleaved probes in the solution are then detected with an LF immunoassay. The real-time assay shows that RIDA is able to specifically detect target sequences in 5 to 15 min. The RIDA-LF combined assay can specifically detect nucleic acid targets without sophisticated equipment. In this report, our data suggest that RIDA is a flexible simple assay that could be applied for point-of-care detection. The modified-RIDA described in this report further extends the application of this technology. PMID- 17911000 TI - An integrated microfluidic system using magnetic beads for virus detection. AB - An integrated system capable of sample pretreatment using antibody-conjugated magnetic beads and one-step reverse transcriptase-polymerase chain reaction (RT PCR) on a microfluidic system was developed to accelerate the detection of RNA viruses such as dengue virus or enterovirus 71. The targeted virus in the sample was first captured by the specific antibody-conjugated magnetic beads, which were manipulated by micro-electromagnets made of micro-electro-mechanical systems technology. The RNA of the targeted virus then underwent thermal lysis and was reverse-transcripted to cDNA using a microRT-PCR module. The sensitivity to detect dengue virus is around 10-100 PFU, which is equivalent to the commercial RNA extraction kit and a large-scale RT-PCR machine. This microsystem can specifically detect 4 serotypes of dengue virus, as well as enterovirus 71. The specificity was warranted by both antibody and primer. The microfluidic system allows automatic process of sample including mixing, incubation, and reaction. The antibody-conjugated magnetic beads offer sample pretreatment of purification and concentration. The integration of antibody-conjugated magnetic beads into the microfluidic system is promising for fast molecular diagnosis of microorganisms. PMID- 17911001 TI - Activity of ceftobiprole against community-associated methicillin-resistant Staphylococcus aureus isolates recently recovered from US military trainees. AB - Ceftobiprole MICs at which 50% and 90% of isolates were inhibited (MIC50 and MIC90), determined by the Clinical and Laboratory Standards Institute broth microdilution method, were both 1 microg/mL (range, 0.5-1 microg/mL) against 143 community-associated methicillin-resistant Staphylococcus aureus isolates and 0.5 microg/mL (range, 0.25-0.5 microg/mL) with 29 methicillin-susceptible isolates recovered from military trainees during 2 prospective investigations. PMID- 17911002 TI - Species-specific diagnostic marker for rapid identification of Staphylococcus aureus. AB - Staphylococcus aureus is a common bacterial pathogen that has emerged as an increasingly important health concern. Following the recent publication of the genome sequences of 9 S. aureus strains (http://www.ncbi.nlm.nih.gov/genomes/lproks.cgi), a gene of S. aureus that relates signal transduction as a target for rapid detection and identification of the pathogen has been investigated. By sequence analysis of S. aureus signal transduction genes from the complete genome of S. aureus ATCC N315 and their comparison with other DNA sequences using GenBank BLAST searches, we identified a unique gene, vicK. Polymerase chain reaction primers (vicK1 and vicK2) derived from this gene allowed amplification of a 289-bp DNA fragment only from S. aureus and not from other Staphylococcus species and other common bacteria tested. Besides offering an additional target for specific confirmation of S. aureus, further analysis of the signal transduction gene vicK and its related protein product may lead to new insights into the molecular mechanisms of S. aureus maintenance and pathogenicity. PMID- 17911003 TI - Assessment of the content of phenolics and antioxidant actions of the Rubiaceae, Ebenaceae, Celastraceae, Erythroxylaceae and Sterculaceae families of Mauritian endemic plants. AB - There is continued interest in the assessment of the bioefficacy of the active principles in extracts from a variety of traditional medicine and food plants in order to determine their impact on the management of a variety of clinical conditions and maintenance of health. The polyphenolic composition and antioxidant potential of Mauritian endemic plants of the Rubiaceae, Ebenaceae, Celastraceae, Erythroxylaceae and Sterculaceae family were determined. The phenolics level of the plant extracts varied from 1 to 75 mg/g FW, the maximum level measured in Diospyros neraudii (Ebenaceae). Coffea macrocarpa showed the highest flavonoids content with 18+/-0.7 mg/g FW. The antioxidant capacity based on the TEAC and FRAP values were strongly related to total phenolics and proanthocyanidins content, while a weaker correlation was observed with (-) gallic acid. Erythroxylum sideroxyloides showed the highest protective effect in the lipid peroxidation systems with IC(50) of 0.0435+/-0.001 mg FW/ml in the Fe(3+)/ascorbate system and 0.05+/-0.002 mg FW/ml in the AAPH system. Cassine orientalis, E. sideroxyloides, Diospyros mellanida and Chassalia coriancea var. johnstonii were weakly prooxidant only at higher concentration greater of 10 g FW/L indicating potential safety. Mauritian endemic plants, particularly the genus Diospyros, are good sources of phenolic antioxidants and potential candidates for the development of prophylactic agents. PMID- 17911004 TI - Human psychoneuroimmunology: 20 years of discovery. AB - An important component of psychoneuroimmunology research is to reveal the myriad ways that behaviors and health are inter-related, with a focus on the immunological mechanisms that underlie these interactions. Research in human psychoneuroimmunology has shown that immunoregulatory processes are an integral part of a complex network of adaptive responses. As such, this review provides a perspective from our laboratory over the last 20 years to define the inter relationships between behavior and immunity; to identify the hypothalamic pituitary adrenal (HPA) and autonomic mechanisms that link the central nervous system and immune responses; to examine the clinical implications of immune alterations during depression or life stress on inflammatory and infectious disease risk; and to explore the reciprocal role of immune mediators on behavior in humans. PMID- 17911006 TI - Thermogravimetric analysis and kinetic study on pyrolysis of representative medical waste composition. AB - To obtain detailed information on the pyrolysis characteristics, a thermogravimetric study on the pyrolysis of 14 typical medical waste compositions was carried out in thermogravimetric analysis (TGA) equipment using dynamic techniques in a stream of N2. An index representing pyrolysis reactivity of waste was presented. Kinetic parameters were obtained by Coats-Redfern method and used to model the TG curve. The results showed that: (a) Plastic, protein, cellulosic material, synthetic fibre, and rubber entered pyrolysis process in succession. (b) There was one decomposition stage in the pyrolysis of one-off medical glove, operating glove, cellulosic waste, absorbable catgut suture and adhesive plaster, while other components had two obvious weight loss stages. (c) The obtained apparent activation energy for second stage pyrolysis was comparably higher than that for first stage. (d) Each stage was controlled by only one kinetic mechanism, in which kinetic parameters were constant. (e) The degradation kinetics of medical waste may be affected by special physical and chemical treatment in the product manufacturing process. (f) Among 13 waste samples, the pyrolysis index of cellulosic matter was the highest, which indicated cellulosic matter had strong pyrolysis reactivity. (g) With increasing heating rate, TG curve and DTG peak shifted to high temperatures and main reaction interval of the sample became longer. PMID- 17911005 TI - Levels of plasma insulin, leptin and adiponectin, and activities of key enzymes in carbohydrate metabolism in skeletal muscle and liver in fasted ICR mice fed dietary n-3 polyunsaturated fatty acids. AB - The aim of this study was to clarify the mechanisms related to plasma glucose concentration in mice fed a diet rich in n-3 polyunsaturated fatty acids (n-3 PUFAs). Male Crlj:CD-1 (ICR) mice were fed experimental diets containing 6% lard (LD), 6% fish oil (FO) or 4.1% lard plus 1.5% docosahexaenoic acid ethyl ester and 0.4% eicosapentaenoic acid ethyl ester (DE) for 12 weeks. There were no marked differences in plasma glucose and insulin concentration changes on glucose tolerance test between the three dietary groups. At the end of the feeding trial, plasma glucose concentration was significantly lower in fasted mice in the FO group than in those in the LD group (P<.005). Plasma adiponectin concentration was significantly higher in the FO group than in the LD group (P<.05). Hexokinase, phosphofructokinase, glucose-6-phosphate dehydrogenase and glycerophosphate dehydrogenase activities in skeletal muscle tended to be lower in the FO group than in the LD group, while there were no differences in glucokinase and phosphofructokinase activities in liver between the three dietary groups. However, hepatic glycerophosphate dehydrogenase activity was 53-fold and 4.2-fold higher in the FO group than in the LD and DE groups, respectively (P<.0005 and P<.05, respectively). These results suggest that the reduction in plasma glucose concentration in mice fed n-3 PUFAs is mainly caused by acceleration of glucose uptake and glycerol synthesis in the liver rather than in the skeletal muscle. PMID- 17911007 TI - Prognostic significance of oestrogen receptor alpha (ERalpha) and beta (ERbeta), progesterone receptor A (PR-A) and B (PR-B) in endometrial carcinomas. AB - The expression of the classic steroid receptors ERalpha and PR-A has been correlated with stage, histological grade and survival in endometrial cancer. Endometrial cancer samples (293) were immunohistochemically analysed with monoclonal antibodies against the four steroid receptors. The loss of ERalpha, PR A and PR-B resulted in a poorer survival in endometrial cancer patients, while ERbeta expression did not demonstrate any correlations with several analysed clinicopathological characteristics and did not affect survival. Additionally, multivariate survival analysis demonstrated that PR-B was a significant independent prognostic factor for cause-specific survival. In contrast, although ERalpha and PR-A showed a significant association between different endometrial histological subtypes and grading, both receptors were not independent factors with survival in endometrial carcinoma patients. Therefore, the PR-B immunostaining might be used as an easy, simple and highly efficient marker to identify high-risk patients and may aid in the selection of patients for a more aggressive adjuvant therapy. PMID- 17911008 TI - Hyaluronan-induced masking of ErbB2 and CD44-enhanced trastuzumab internalisation in trastuzumab resistant breast cancer. AB - Although trastuzumab, a recombinant humanised anti-ErbB2 antibody, is widely used in the treatment of breast cancer, neither its mechanism of action, nor the factors leading to resistance are fully understood. We have previously shown that antibody-dependent cellular cytotoxicity is pivotal in the in vivo effect of trastuzumab against JIMT-1, a cell line showing in vitro resistance to the antibody, and suggested that masking of the trastuzumab-binding epitope by MUC-4, a cell surface mucin, took place. Here, we further explored the role of masking of ErbB2 in connection with CD44 expression and synthesis of its ligand, hyaluronan. We show that high expression of CD44 observed in JIMT-1 cells correlates with ErbB2 downregulation in vivo, while siRNA-mediated inhibition of CD44 expression leads to decreased rate of trastuzumab internalisation and low cell proliferation in vitro. An inhibitor of hyaluronan synthesis, 4 methylumbelliferon (4-MU) significantly reduced the hyaluronan level of JIMT-1 cells both in vivo and in vitro leading to enhanced binding of trastuzumab to ErbB2 and increased ErbB2 down-regulation. Furthermore, the inhibitory effect of trastuzumab on the growth of JIMT-1 xenografts was significantly increased by 4 MU treatment. Our results point to the importance of the CD44-hyaluronan pathway in the escape of tumour cells from receptor-oriented therapy. PMID- 17911009 TI - beta-Carotene induces apoptosis and up-regulates peroxisome proliferator activated receptor gamma expression and reactive oxygen species production in MCF 7 cancer cells. AB - Although the pharmacological role of beta-carotene in the prevention and treatment of many cancer cells has received increasing attention, the molecular mechanisms underlying such chemopreventive activity are not clear. Since peroxisome proliferator-activated receptor gamma (PPAR-gamma) has been implicated in regulating breast cancer cell differentiation and apoptosis, the effects of beta-carotene on the PPAR-gamma-mediated pathway and its association with reactive oxygen species production in MCF-7 cells were investigated in the present study. The results demonstrated that beta-carotene significantly increased PPAR-gamma mRNA and protein levels in time-dependent manner. In addition, beta-carotene increased the cyclin-dependent kinase inhibitor p21(WAF1/CIP1) expression and decreased the prostanoid synthesis rate-limiting enzyme cyclooxygenase-2 expression. 2-chloro-5-nitro-N-phenylbenzamide (GW9662), an irreversible PPAR-gamma antagonist, partly attenuated the cell death caused by beta-carotene. Further, reactive oxygen species (ROS) production was induced by beta-carotene, resulting in mitochondrial dysfunction and cytochrome C release. Reduced glutathione (GSH) treatment decreases the intracellular ROS and prevents cytochrome C release and cell apoptosis induced by beta-carotene. In total, these observations suggest that the synergistic effect of PPAR-gamma expression and ROS production may account for beta-carotene-mediated anticancer activities. PMID- 17911010 TI - Production of fungal biomass protein using microfungi from winery wastewater treatment. AB - This study was carried out to investigate the production of fungal biomass protein (FBP) in treatment of winery wastewater using microfungi. Three fungal strains, Trichoderma viride WEBL0702, Aspergillus niger WEBL0901 and Aspergillus oryzae WEBL0401, were selected in terms of microbial capability for FBP production and COD reduction. T. viride appeared to be the best strain for FBP production due to high productivity and less nitrogen requirement. More than 5 g/L of fungal biomass was produced in shake fermentation using T. viride without nitrogen addition, and by A. oryzae and A. niger with addition of 0.5-1.0 g/L (NH4)2SO4. The FBP production process corresponded to 84-90% COD reduction of winery wastewater. Fungal biomass contained approximately 36% protein produced by two Aspergillus strains, while biomass produced by T. viride consisted of 19.8% protein. Kinetic study indicated that maximum fungal cell growth could be achieved in 24h for T. viride and 48 h for A. oryzae and A. niger. Current results indicated that it could be feasible to develop a biotechnological treatment process integrated with FBP production from the winery waste streams. PMID- 17911012 TI - An evaluation of the phosphorus storage capacity of an anaerobic/aerobic sequential batch biofilm reactor. AB - The aim of this work was to assess the phosphorus storage capability of the polyphosphate (poly-P) accumulating organisms (PAO) in the biofilm using a sequential batch biofilm reactor (SBBR). In the anaerobic phase, the specific COD uptake rates increases from 0.05 to 0.22 (mg-COD/mg-biomass/h) as the initial COD increases and the main COD uptake activity occurs in the initial 30 min. The polyhydroxyalkanoates (PHAs) accumulation from 18 to 38 (mg-PHA/g-biomass) and phosphorus release from 20 to 60 (mg-P/L) share a similar trend. The adsorbed COD cannot be immediately transformed to PHAs. Since the PHAs' demand per released phosphorus is independent of the initial COD, the enhancement of the PHA accumulation would be of benefit to phosphorus release. The only requirement is to have an initial amount of substrate that will result in sufficient PHA accumulation (approximately 20 mg-PHA/g-biomass) for phosphorus release. During the aerobic phase, the aeration should not only provide sufficient dissolved oxygen, but should also enhance the mass transfer and the diffusion. In other words, the limitation to the phosphorus storage capability always occurs during the anaerobic phase, not the aerobic phase. PMID- 17911013 TI - Inactivation of ANAMMOX communities under concurrent operation of anaerobic ammonium oxidation (ANAMMOX) and denitrification. AB - A concurrent operation of anaerobic ammonium oxidation (ANAMMOX) and denitrification was investigated in a well known UASB reactor seeding with both ANAMMOX and anaerobic granular sludges. ANAMMOX activity was confirmed by hydroxylamine test and the hybridization of biomass using the gene probes of Amx 820 and EUB 338 mixed. Denitrification was observed through the reductions of both COD and nitrate-nitrite concentrations under anaerobic/anoxic conditions. By providing a stoichiometric ratio of nitrite to ammonium nitrogen with addition nitrate nitrogen, a gradual reduction of ANAMMOX activity was found with an increase of COD concentration in a range of 100-400 mg l(-1). This is equivalent to the COD to N ratio of 0.9-2.0. The COD concentration was found to be a control variable for process selection between ANAMMOX reaction and denitrification. A reduction of COD and nitrite-nitrate concentrations in all reactors confirmed the undergone concurrent denitrification which thrives when sufficient organic matter is available. COD concentration over 300 mg l(-1) was found to inactivate or eradicate ANAMMOX communities. PMID- 17911011 TI - Effect of heat-generated product from uronic acids on the physiological activities of microbial cells and its application. AB - Filtered samples of monogalacturonic (GA) and monoglucuronic acids (GL) that were prepared using millipore filter (pore size=0.2 microm) slightly inhibited the growth of Escherichia coli while the autoclaved (at 121 degrees C for 20 min) samples of GA and GL completely inhibited the growth of E. coli. The most effective substance generated upon autoclave treatment was isolated and characterized as trans-4,5-dihydroxy-2-cyclopenten-1-one (DHCP). The optimal conditions for DHCP generation were also established by autoclaving GA (pH 2.3) at 121 degrees C for 3h. DHCP completely inhibited the growth of E. coli. However, the growth of E. coli was restored when superoxide dismutase and catalase were added to the culture broth that contained DHCP. It was thought that DHCP might have induced the release of active oxygen, which resulted in the inhibition of microbial growth. In the case of gram-positive bacteria (Bacillus cereus, Bacillus subtilis and Staphylococcus aureus) and yeast (Saccharomyces cerevisiae and Candida brassicae), DHCP inhibited the cell growth. Based on our results, methods for preparation of food preservatives that contained pectin degraded products (oligo-galacturonic acid and monogalacturonic acid) and DHCP were developed. The preservatives were very effective in inhibiting the growth of E. coli and S. cerevisiae. PMID- 17911014 TI - Effect of matured compost as a bulking and inoculating agent on the microbial community and maturity of cattle manure compost. AB - Cattle manure composts were consecutively manufactured. Compost that reached maturity first was used as a bulking and inoculating agent for subsequent compost production. The microbial community was measured through phospholipid fatty acid analysis. Changes in the content of fatty acid methyl esters derived from phospholipids were similar in all the composts. The diversity index for the fatty acid methyl ester content increased in the secondary-produced compost from the onset of composting. Microbial succession was accelerated using matured compost. The proportion of biomarker fatty acids for gram-positive bacteria also increased in the secondary-produced compost from the early stage of composting. Changes in germination index indicated the maturity stage of the compost. The proportion of biomarker fatty acids for gram-positive bacteria was positively correlated to the germination index, indicating that phospholipid fatty acid analysis is an indicator for evaluating the maturity of cattle manure composts. PMID- 17911015 TI - Volumetric scale-up of a three stage fermentation system for food waste treatment. AB - In this study, a volumetric scale-up of this system was designed and built on a field pilot-scale (total digester volume 10 m(3)), with the results from the field pilot-scale experiments compared with those from the bench-scale (total digester volume 0.4 m(3)) process prior to scale-up. The reduction rate of total chemical oxygen demand (tCOD) and the maximum methane content produced in the biogas from the bench-scale system were 90.6% and 72%; whereas those from the field pilot-scale system were 90.1% and 68%, respectively. The estimated methane yields were 282 and 254 l CH(4)/kg tCOD(degraded) in bench and field pilot-scale fermentation systems, respectively. These results indicate that the three stage fermentation system developed in this study can be applied as a commercial process for the disposal of food waste in view of process stability. PMID- 17911016 TI - Extraction of astaxanthin from giant tiger (Panaeus monodon) shrimp waste using palm oil: studies of extraction kinetics and thermodynamic. AB - Study of extraction of astaxanthin from giant tiger (Panaeus monodon) shrimp waste using palm oil was conducted to determine the extraction kinetics and thermodynamic parameters. Two extraction models were proposed: mass transfer kinetic model and reaction kinetic model. It was found that both of mass transfer and reaction kinetic control the extraction of astaxanthin from shrimp waste using palm oil. The thermodynamic parameters of extraction were also obtained in this study. PMID- 17911017 TI - Microbial transformation of three bufadienolides by Nocardia sp. and some insight for the cytotoxic structure-activity relationship (SAR). AB - Resibufogenin, cinobufagin, and bufalin are cytotoxic steroids isolated from the Chinese drug Chan'su. Biotransformation of these three bufadienolides by Nocardia sp. NRRL 5646 was investigated. Notably, resibufogenin was converted to 3-acetyl 15beta-hydroxyl bufotalin, via an unprecedented 14beta,15beta-epoxy ring cleavage and a regio-selective acetoxylation. This product showed significantly increased cytotoxic activity. The regio-selective acetylation of the 3-OH was also involved in the other reactions. The structures of metabolites were established by ESI LC/MS and 2D NMR techniques. The in vitro cytotoxic activities against human cancer cell lines of the substrates and the transformed products were determined by the MTT method and their structure-activity relationship (SAR) was discussed. This investigation provided a useful approach to prepare new bufadienolides and the SAR research. PMID- 17911018 TI - 3,5-bis(3'-indolyl)pyrazoles, analogues of marine alkaloid nortopsentin: synthesis and antitumor properties. AB - A series of 10 bis-indolylpyrazoles of type 9, 10 were obtained by cyclization of diketones 8 using hydrazine monohydrate or methylhydrazine in refluxing acetic acid/THF. Derivatives 9a,c,d were selected, by the National Cancer Institute (NCI, Bethesda, USA), to be evaluated against the full panel of about 60 human tumor cell lines derived from nine human cancer cell types and showed antiproliferative activity in the micromolar range. In particular, 9d, the most active compound was effective against all the tested cell lines with a GI(50) mean value of 3.23 microM; TGI and LC(50) values were 14.5 and 58.9 microM having positive response on 91% and 41% of the tested cell lines, respectively. PMID- 17911019 TI - Novel scaffold for cathepsin K inhibitors. AB - Pyrrolopyrimidine, a novel scaffold, allows to adjust interactions within the S3 subsite of cathepsin K. The core intermediate 10 facilitated the P3 optimization and identified highly potent and selective cathepsin K inhibitors 11-20. PMID- 17911020 TI - Novel synthetic ligands for targeted PET imaging and radiotherapy of copper. AB - Novel ligands, NBEA, NBPA, NETA, NE3TA, and NE3TA-Bn, were synthesized and evaluated as potential chelators of copper radioisotopes for use in targeted positron emission tomography (PET) imaging or radiation therapy. The new ligands were radiolabeled with (64)Cu, and in vitro stability of the radiolabeled complexes was assessed in rat serum. Serum stability results suggest that among the ligands tested, NETA, NE3TA, and NE3TA-Bn form stable complexes with (64)Cu. PMID- 17911021 TI - Substituted 2-pyrrolinone inhibitors of HIV-1 integrase. AB - The beta-diketoacid class of HIV-1 integrase (IN) inhibitors represent the first potent class of compounds specific for the strand transfer catalytic activity of the viral enzyme. Previously, utilizing a beta-diketoacid pharmacophore as a search query, we identified a substituted 2-pyrrolinone with modest IN inhibitory activity from a database of small-molecules [Dayam, R.; Sanchez, T.; Neamati, N. J. Med. Chem.2005, 48, 8009]. In efforts to optimize this class of IN inhibitors, we carried out a structure-activity relationship analysis around the 2 pyrrolinone core. Here, we present a new class of 2-pyrrolinone IN inhibitors. PMID- 17911022 TI - Discovery of novel 8-azoniabicyclo[3.2.1]octane carbamates as muscarinic acetylcholine receptor antagonists. AB - In the course of our research program to develop novel muscarinic receptor antagonists for the treatment of COPD, new tropane carbamate derivatives were identified as potent anti-muscarinic agents. The synthesis, structure-activity relationships and pharmacological evaluation that led to the identification of compound 5o, are described. PMID- 17911023 TI - 3,5-Disubstituted quinolines as novel c-Jun N-terminal kinase inhibitors. AB - The structure-based design and synthesis of a novel series of c-Jun N-terminal kinase (JNK) inhibitors with selectivity against p38 is reported. The unique structure of 3,5-disubstituted quinolines (2) was developed from the previously reported 4-(2,7-phenanthrolin-9-yl)phenol (1). The X-ray crystal structure of 16a in JNK3 reveals an unexpected binding mode for this new scaffold with protein. PMID- 17911024 TI - Potent oxindole based human beta3 adrenergic receptor agonists. AB - The synthesis and biological evaluation of a series of oxindole beta(3) adrenergic receptor agonists is described. A modulation of rat atrial tachycardia was observed with substitution at the 3-position of the oxindole moiety. PMID- 17911025 TI - Prospective audit of emergency department transit times associated with entonox analgesia for reduction of the acute, traumatic dislocated shoulder. AB - AIM: To establish current UK practice for the management of the acute traumatic shoulder dislocation with respect to analgesia and reduction manoeuvres. To compare the transit times of patients through an emergency department (ED) after the use of intravenous analgesia and/ or sedation compared to entonox +/- simple oral analgesia. METHODS: A postal questionnaire was sent to 100 UK ED consultants to establish current practice. The treating clinicians were allowed to choose the method of analgesia provided to reduce the patient's dislocated shoulder, provided the patient was happy with it. They administered either (1) traditional intravenous morphine and/or midazolam or (2) entonox +/- simple oral analgesia to facilitate reduction. A prospective audit was conducted to compare the transit times of the two groups of patients. RESULTS: The postal questionnaire revealed that intravenous morphine and midazolam are widely used during reduction of the acute shoulder dislocation in the UK. The audit showed that this was associated with a significantly prolonged transit time through the ED, compared to entonox alone, (mean 77 min versus 177 min, respectively, p<0.001) without compromise in reduction success. CONCLUSION: Entonox +/- simple oral analgesia significantly decreases ED transit times as compared to IV morphine and/or midazolam for the reduction of the acute traumatic dislocated shoulder. Further studies should be done into patient pain scores and into the best combination of oral analgesia and entonox. PMID- 17911026 TI - 1, 25 Dihydroxyvitamin D3 modulated cytokine response in pulmonary tuberculosis. AB - 1, 25 Dihydroxyvitamin D(3) (1, 25(OH)(2) D(3)) has gained significant importance in tuberculosis with regard to its immunoregulatory activities. Our aim was to evaluate the effect of 1, 25(OH)(2) D(3) on cytokine response to Mycobacterium tuberculosis antigens in pulmonary tuberculosis. Peripheral blood mononuclear cells obtained from 60 healthy controls and 52 pulmonary tuberculosis patients were cultured with culture filtrate antigen (CFA) of M. tuberculosis and live M. tuberculosis with and without 1, 25(OH)(2) D(3) (10(-9), 10(-8)and 10(-7)M concentrations). The culture supernatants were used to estimate IL-8, IL-6, TGF beta, IL-10, IFN-gamma, IL-12p40, IL-2, IL-4 and IL-5 levels by ELISA. 1, 25 Dihydroxyvitamin D(3) significantly suppressed IL-12p40 and IFN-gamma production in response to CFA and live M. tuberculosis with a maximum suppression at 10(-7)M concentration (p<0.0001). In CFA stimulated cultures, addition of 1, 25(OH)(2) D(3) significantly suppressed IL-8, IL-6 and IL-10 whereas the IL-2 levels were significantly increased in controls. It variably influenced the Th2 cytokines, showing an increased trend for IL-4 and suppressed IL-5 levels. We report that 1, 25(OH)(2) D(3) differentially modulates production of cytokines in response to M. tuberculosis antigens by predominantly suppressing IL-12p40 and IFN-gamma production in a dose dependent manner. Our results suggest a role for vitamin D in restricting acquired immune response against tuberculosis by regulating cytokine production. PMID- 17911027 TI - Spontaneous fibrillation of the native neuropeptide hormone Somatostatin-14. AB - Natural Somatostatin-14 is a small cyclic neuropeptide hormone with broad inhibitory effects on endocrine secretions. Here we show that natural Somatostatin-14 spontaneously self-assembles in water and in 150 mM NaCl into liquid crystalline nanofibrils, which follow characteristic structural features of amyloid fibrils. These non-covalent highly stable structures are based on the Somatostatin native backbone conformation and are formed under non-denaturing conditions. Our results support the hypothesis that self-assembly into amyloid fibrils is a generic property of the polypeptide chain under appropriate conditions. Given recent advances on the mechanisms of biological storage and sorting modes of peptide/protein hormones into secretory granules, we propose that Somatostatin-14 fibrillation could be relevant to the regulated secretion pathway of this neuropeptide hormone. Such a hypothesis is consistent with the emerging concept of the existence of non-disease related but functional amyloids. PMID- 17911028 TI - Fast, robust, and accurate determination of transmission electron microscopy contrast transfer function. AB - Transmission electron microscopy, as most imaging devices, introduces optical aberrations that in the case of thin specimens are usually modeled in Fourier space by the so-called contrast transfer function (CTF). Accurate determination of the CTF is crucial for its posterior correction. Furthermore, the CTF estimation must be fast and robust if high-throughput three-dimensional electron microscopy (3DEM) studies are to be carried out. In this paper we present a robust algorithm that fits a theoretical CTF model to the power spectrum density (PSD) measured on a specific micrograph or micrograph area. Our algorithm is capable of estimating the envelope of the CTF which is absolutely needed for the correction of the CTF amplitude changes. PMID- 17911029 TI - Influence of gravity compensation on muscle activity during reach and retrieval in healthy elderly. AB - INTRODUCTION: Arm support like gravity compensation may improve arm movements during stroke rehabilitation. It is unknown how gravity compensation affects muscle activation patterns during reach and retrieval movements. Since muscle activity during reach is represented by a component varying with movement velocity and a component supposedly counteracting gravity, we hypothesized that gravity compensation decreases the amplitude of muscle activity, but does not affect the pattern. To examine this, we compared muscle activity during well defined movements with and without gravity compensation in healthy elderly. METHODS: Ten subjects performed reach and retrieval movements with and without gravity compensation. Muscle activity of biceps, triceps, anterior, middle and posterior parts of deltoid and upper trapezius was compared between the two conditions. RESULTS: The level of muscle activity was lower with gravity compensation in all muscles, reaching significance in biceps, anterior deltoid and trapezius (p < or = 0.026). The muscle activation pattern did not differ between movements with and without gravity compensation (p > or = 0.662). DISCUSSION: Gravity compensation only influenced the level of muscle activity but not the muscle activation pattern in terms of timing. Future studies should examine if the influence of gravity compensation is comparable for stroke patients. This may stimulate early and intensive training during rehabilitation. PMID- 17911030 TI - Probabilistic streamline q-ball tractography using the residual bootstrap. AB - Q-ball imaging has the ability to discriminate multiple intravoxel fiber populations within regions of complex white matter architecture. This information can be used for fiber tracking; however, diffusion MR is susceptible to noise and multiple other sources of uncertainty affecting the measured orientation of fiber bundles. The proposed residual bootstrap method utilizes a spherical harmonic representation for high angular resolution diffusion imaging (HARDI) data in order to estimate the uncertainty in multimodal q-ball reconstructions. The accuracy of the q-ball residual bootstrap technique was examined through simulation. The residual bootstrap method was then used in combination with q ball imaging to construct a probabilistic streamline fiber tracking algorithm. The residual bootstrap q-ball fiber tracking algorithm is capable of following the corticospinal tract and corpus callosum through regions of crossing white matter tracts in the centrum semiovale. This fiber tracking algorithm is an improvement upon prior diffusion tensor methods and the q-ball data can be acquired in a clinically feasible time frame. PMID- 17911031 TI - Reproducible activation in BA2, 1 and 3b associated with texture discrimination in healthy volunteers over time. AB - We aimed to quantify specific location and reproducibility of brain activation associated with discrimination of a moving textured surface in adult healthy volunteers over a 6-month interval. A sensory stimulation device was developed to provide a texture stimulus to the fingertips at a controlled speed and pressure. Repeat measurements of regional cerebral blood flow, using positron emission tomography (PET), were obtained in 10 healthy individuals, aged 33 to 80 years (mean=55.8 years), at scanning sessions separated by 6 months. Stimulation and rest conditions were presented to either the right, dominant (n=5) or left non dominant (n=5) hand. Activation location was objectively quantified with reference to probabilistic cytoarchitectonic maps. Differences in activation over time and regions of common activation were also quantified. Participants consistently activated Brodmann areas (BA) 2, 3b and 1, somatosensory areas of postcentral gyrus, at initial and 6-month studies: 93.1% of common activation for the right-hand (RH) and 60.6% for left-hand (LH) stimulation group were in these areas. Reproducible activation in BA6, 4a and 4p was also observed for the RH group (6.8% of common activation) and LH group (39.4%). There were no sites of significant difference over time for either hand. Highly consistent location of activation over time suggests that changes in loci of activation may be confidently monitored in adults using this paradigm. Use of probabilistic cytoarchitectonic maps permitted objective quantification of the anatomical location of the core of reproducible activation. PMID- 17911033 TI - Phylogeny of pteromalid parasitic wasps (Hymenoptera: Pteromalidae): initial evidence from four protein-coding nuclear genes. AB - Chalcidoidea (approximately 22,000 described species) is the most ecologically diverse superfamily of parasitic Hymenoptera and plays a major role in the biological control of insect pests. However, phylogenetic relationships both within and between chalcidoid families have been poorly understood, particularly for the large family Pteromalidae and relatives. Forty-two taxa, broadly representing Chalcidoidea but concentrated in the 'pteromalid lineage,' were sequenced for 4620 bp of protein-coding sequence from four nuclear genes for which we present new primers. These are: CAD (1719 bp) DDC (708 bp), enolase (1149 bp), and PEPCK (1044 bp). The combined data set was analyzed using parsimony, maximum likelihood, and Bayesian methods. Statistical significance of the apparent non-monophyly of some taxonomic groups on our trees was evaluated using the approximately unbiased test of Shimodaira [Shimodaira, H. 2002. An approximately unbiased test of phylogenetic tree selection. Syst. Biol. 51(3), 492-508]. In accord with previous studies, we find moderate to strong support for monophyly of Chalcidoidea, a sister-group relationship of Mymaridae to the remainder of Chalcidoidea, and a relatively basal placement of Encarsia (Aphelinidae) within the latter. The 'pteromalid lineage' of families is generally recovered as monophyletic, but the hypothesis of monophyly for Pteromalidae, which appear paraphyletic with respect to all other families sampled in that lineage, is decisively rejected (P < 10(-14)). Within Pteromalidae, monophyly was strongly supported for nearly all tribes represented by multiple exemplars, and for two subfamilies. All other multiply-represented subfamilies appeared para- or polyphyletic in our trees, although monophyly was significantly rejected only for Miscogasterinae, Ormocerinae, and Colotrechninae. The limited resolution obtained in the analyses presented here reinforces the idea that reconstruction of pteromalid phylogeny is a difficult problem, possibly due to rapid radiation of many chalcidoid taxa. Initial phylogenetic comparisons of life history traits suggest that the ancestral chalcidoid was small-bodied and parasitized insect eggs. PMID- 17911032 TI - Motor and language DTI Fiber Tracking combined with intraoperative subcortical mapping for surgical removal of gliomas. AB - Preoperative DTI Fiber Tracking (DTI-FT) reconstruction of functional tracts combined with intraoperative subcortical mapping (ISM) is potentially useful to improve surgical procedures in gliomas located in eloquent areas. Aims of the study are: (1) to evaluate the modifications of fiber trajectory induced by the tumor; (2) to validate preoperative DTI-FT results with intraoperative identification of functional subcortical sites through direct subcortical stimulation; (3) to evaluate the impact of preoperative DTI-FT reconstructions in a neuronavigational setup combined with ISM technique on duration and modalities of surgical procedures, and on functional outcome of the patients. Data are available on 64 patients (52 low-grade and 12 high-grade gliomas). DTI-FT was acquired by a 3-T MR scanner with a single-shot EPI sequence (TR/TE 8986/80 ms, b=1000 s/mm) with gradients applied along 32 non-collinear directions. 3D Fast Field Echo (FFE) T1-weighted imaging (TR/TE 8/4 ms) was performed for anatomic guidance. The corticospinal tract (CST), superior longitudinal, inferior fronto occipital and uncinatus fasciculi were reconstructed. Data were transferred to the neuronavigational system. Functional subcortical sites identified during ISM were correlated with fiber tracts depicted by DTI-FT. In high-grade gliomas, DTI FT depicted tracts mostly at the tumor periphery; in low-grade gliomas, fibers were frequently located inside the tumor mass. There was a high correlation between DTI-FT and ISM (sensitivity for CST=95%, language tracts=97%). For a proper reconstruction of the tracts, it was necessary to use a low FA threshold of fiber tracking algorithm and to position additional regions of interest (ROIs). The combination of DTI-FT and ISM decreased the duration of surgery, patient fatigue, and intraoperative seizures. Combination of DTI-FT and ISM allows accurate identification of eloquent fiber tracts and enhances surgical performance and safety maintaining a high rate of functional preservation. PMID- 17911034 TI - Simultaneous polyphenism and cryptic species in an intertidal limpet from New Zealand. AB - The small intertidal limpets known under the name Notoacmea helmsi occupy a wide variety of habitats in New Zealand and exhibit a variety of shell forms. Phylogenetic analyses of DNA sequences from two genes, mitochondrial COI and nuclear ITS1, reveal that this taxon comprises at least five morphologically cryptic species, with at least one of these species, N. scapha, consisting of individuals with two obviously different shell types. One of these forms is an ecophenotypic response to living on eelgrass (Zostera) fronds. Unlike its extinct relative, Lottia alveus, N. scapha is not restricted to this substrate, but individuals living elsewhere are larger and have a different shell shape. Although there is significant overlap in shell form among the different cryptic species, there is some habitat differentiation, with two species predominantly found on exposed shores and three confined to mudflats. One species exhibits distinctive light-avoiding behaviour, the first known case in which behaviour can be used to separate cryptic species in molluscs. PMID- 17911035 TI - Comparative phylogeography of three skink species (Oligosoma moco, O. smithi, O. suteri; Reptilia: Scincidae) in northeastern New Zealand. AB - Sea-level fluctuations during the Pliocene and Pleistocene have shaped the landscape of the Northland region of New Zealand. We examined the comparative phylogeography of three skink species (Oligosoma moco, O. smithi, O. suteri) in northeastern New Zealand in order to investigate the impact of the historical processes that have prevailed since the Pliocene on the Northland fauna. O. moco, O. smithi and O. suteri have similar distributions across northeastern New Zealand, frequently occurring in sympatry. We obtained sequence data from across the entire range of each species, targeting the ND2 mitochondrial gene. Using Neighbor-Joining, Maximum likelihood and Bayesian methods, our analysis revealed contrasting phylogeographic patterns in each species. We found substantial phylogeographic structure within O. moco, with three distinct clades identified. Similarly, deep phylogeographic divergence was evident within O. smithi, with three distinct clades present. Clade 1 included O. smithi populations from the Three Kings Islands and the western coastline of Northland, while Clade 2 encompassed the remainder of the range. However, since Clade 3 corresponded to a described species (O. microlepis), O. smithi might represent a species complex. In both O. moco and O. smithi, divergences among clades are estimated to have occurred in the Pliocene, with divergences within clades occurring during the Pleistocene. In contrast, genetic divergence among O. suteri populations was extremely limited and indicative of more recent divergences during the Pleistocene. The lack of phylogeographic structure in O. suteri might be a consequence of its oviparous reproductive mode, which restricted its distribution to warm northern refugia during glacial maxima. Differences in the ecology and biology of each species might have produced contrasting responses to the same historical processes, and ultimately diverse phylogeographic patterns. Our study reveals an absence of consistent and concordant phylogeographic patterns in the Northland biota, even within the same taxonomic group. PMID- 17911036 TI - Immediate as well as delayed post learning sleep but not wakefulness enhances declarative memory consolidation in children. AB - While there is mounting evidence for the importance of sleep for declarative memory consolidation in adults, so far this issue has not been investigated in children despite considerable differences in sleep duration and sleep architecture between children and adults. Here, 27 children (aged between 9 and 12yr) were examined on two conditions: on the Sleep-Wake condition, subjects learned word pairs in the evening and delayed recall was tested first in the next morning after sleep and then again in the following evening after daytime wakefulness. On the Wake-Sleep condition, learning took place in the morning and delayed recall was tested in the evening of the same day and again in the next morning after sleep. In both conditions retention of declarative memory was significantly increased only after an interval of sleep that either followed immediately after learning (as in the Sleep-Wake condition) or that followed after daytime wakefulness (as in the Wake-Sleep condition), respectively. The results support the hypothesis that sleep plays an active role in declarative memory consolidation even if delayed and further show for the first time the importance of sleep for declarative memory consolidation during childhood. PMID- 17911037 TI - The contribution of the synovium, synovial derived inflammatory cytokines and neuropeptides to the pathogenesis of osteoarthritis. AB - Osteoarthritis (OA) is one of the most common and disabling chronic joint disorders affecting horses, dogs and humans. Synovial inflammation or synovitis is a frequently observed phenomenon in osteoarthritic joints and contributes to the pathogenesis of OA through formation of various catabolic and pro inflammatory mediators altering the balance of cartilage matrix degradation and repair. Catabolic mediators produced by the inflamed synovium include pro inflammatory cytokines, nitric oxide, prostaglandin E(2) and several neuropeptides, which further contribute to the pathogenesis of OA by increasing cartilage degradation. Recent studies suggest that substance P, corticotropin releasing factor, urocortin and vasoactive intestinal peptide may also be involved in OA development, but the precise role of these neuropeptides in the pathogenesis of OA is not known. Since increased production of matrix metalloproteinases by the synovium is stimulated by pro-inflammatory cytokines, future anti-inflammatory therapies should focus on the synovium as a means of controlling subsequent inflammatory damage. PMID- 17911038 TI - Pig-MAP and haptoglobin concentration reference values in swine from commercial farms. AB - Pig-MAP (Major Acute-phase Protein) and haptoglobin concentrations were determined in pigs from commercial farms, and reference intervals obtained for different productive stages. Pig-MAP serum concentrations were lower in sows than in adult boars (mean values 0.81 vs. 1.23 mg/mL) and the opposite was observed for haptoglobin (1.47 vs. 0.94 mg/mL). No differences were found between parities, except for a minor decrease in haptoglobin concentration in the 4th parity. A linear correlation between pig-MAP and haptoglobin concentration was observed. In the period 4-12 weeks of life, pig-MAP mean concentrations were around 1mg/mL, being lower in the finishing period (0.7-0.8 mg/mL). Haptoglobin concentrations increased with time, from around 0.6 mg/mL at 4 weeks of age to 1.4 mg/mL at 12 weeks. Mean values of around 0.9 mg/mL were observed in the finishing period. A wider distribution of values was observed for haptoglobin than for pig-MAP concentrations. Differences between herds were observed, with the highest values obtained in a herd with signs of respiratory disease. PMID- 17911039 TI - An Alu-mediated rearrangement causing a 3.2kb deletion and a novel two base pair deletion in AAAS gene as the cause of triple A syndrome. AB - Triple A syndrome is an autosomal recessive disorder resulting from deleterious mutations in the AAAS gene located on chromosome 12q13. Typical clinical presentation of this syndrome includes adrenal insufficiency, achalasia, and alacrima. A 10-year-old female was diagnosed with Triple A syndrome at the age of 1 year. Initial analysis of the AAAS gene revealed apparently homozygosity for a novel 2bp deletion in exon 1. The father of the patient was heterozygous for this mutation but the mother and the maternal grandparents were apparently homozygous for the wild-type. Further studies demonstrated that the patient carried an intragenic 3.2kb deletion within both 5' and 3' breakpoints located within Alu repeats. The deletion includes 5'-flanking region, exon 1, intron 1, exon 2, and part of intron 2 sequences of the AAAS gene. This Alu-mediated deletion was inherited from her mother and maternal grandmother. This is the first report that Alu-mediated rearrangement in conjunction with a novel two-bp deletion of the AAAS gene is a cause of Triple A syndrome. The results of our study lead to the hypothesis that an Alu-mediated mechanism may be responsible for large alterations in the AAAS gene. We also stress the importance of studying the family in genetic recessive diseases, such as Triple A syndrome, to avoid incorrect diagnosis and to provide accurate genetic counseling. PMID- 17911040 TI - Effects of stimulant medications on the EEG of girls with Attention Deficit/Hyperactivity Disorder. AB - OBJECTIVE: Stimulant medications are the most commonly used treatments for Attention-Deficit/Hyperactivity Disorder (AD/HD) in North America and Australia, although it is still not entirely known how these medications work. This study investigated the effects of stimulant medications on the EEG of girls with AD/HD. METHODS: An initial EEG was recorded during an eyes-closed resting condition. Data from 19 electrode sites were Fourier transformed to provide absolute and relative power estimates for the delta, theta, alpha and beta bands. The data were then averaged into 9 regions and an analysis of both global and regional differences was performed. Subjects were placed on a six-month trial of a stimulant and a second EEG was recorded at the end of the trial. RESULTS: The unmedicated girls had significantly greater total power, absolute delta and theta, more relative theta especially in the frontal regions, and reduced frontal relative delta and beta activity compared with controls. Medication resulted in normalisation of theta power, but after medication, increased relative beta was also apparent in the AD/HD group. CONCLUSIONS: These results indicate that stimulant medications result in a normalisation of slow wave activity in the EEG. In line with published research on the effects of arousal on the EEG, these results suggest that stimulant medications may have their therapeutic effect by improving the EEG substrate of processing deficits in these children. However, this requires further testing during active processing tasks. SIGNIFICANCE: This is the first study to investigate the effect of stimulant medications on the EEG of girls with AD/HD. PMID- 17911041 TI - Executive functions processed in the frontal and lateral temporal cortices: intracerebral study. AB - OBJECTIVE: The study was designed to investigate the neurocognitive network in the frontal and lateral temporal cortices that is activated by the complex cognitive visuomotor tasks of letter writing. METHODS: Eight epilepsy surgery candidates with implanted intracerebral depth electrodes performed two tasks involving the writing of single letters. The first task consisted of copying letters. In the second task, the patients were requested to write any other letter. The cognitive load of the second task was increased mainly by larger involvement of the executive functions. The task-related ERD/ERS of the alpha, beta and gamma rhythms was studied. RESULTS: The alpha and beta ERD as the activational correlate of writing of single letters was found in the sensorimotor cortex, anterior cingulate, premotor, parietal cortices, SMA and the temporal pole. The alpha and beta ERD linked to the increased cognitive load was present moreover in the dorsolateral and ventrolateral prefrontal cortex, orbitofrontal cortex and surprisingly also the temporal neocortex. Gamma ERS was detected mostly in the left motor cortex. CONCLUSIONS: Particularly the temporal neocortex was activated by the increased cognitive load. SIGNIFICANCE: The lateral temporal cortex together with frontal areas forms a cognitive network processing executive functions. PMID- 17911042 TI - EEG differences between eyes-closed and eyes-open resting conditions. AB - OBJECTIVE: Recent work has attempted to clarify the energetics of physiological responding and behaviour by refining and separating the operational definitions of "arousal" and "activation", which have different effects on physiological responding and behaviour. At the EEG level, we relate the former to widespread activity, and the latter to task-specific topographically-focussed activity reflecting regional processing. This study aimed to investigate this further in terms of differences in EEG activity between eyes-closed and eyes-open resting conditions. METHODS: EEG activity was recorded from 28 university students during both eyes-closed and eyes-open resting conditions, Fourier transformed to provide estimates for absolute power in the delta, theta, alpha and beta bands, and analysed in 9 regions across the scalp. Skin conductance level was also measured as an indicator of arousal level. RESULTS: Across the eyes-closed conditions, skin conductance levels were negatively correlated with mean alpha levels. Skin conductance levels increased significantly from eyes-closed to eyes-open conditions. Reductions were found in across-scalp mean absolute delta, theta, alpha and beta from the eyes-closed to eyes-open condition. Topographic changes were also evident in all bands except for alpha, with reduced lateral frontal delta and posterior theta, and decreased posterior/increased frontal beta in the eyes-open condition. In particular, the topographic beta effects indicate that the across-scalp reduction arose from focal reductions rather than global changes. CONCLUSIONS: The obtained results confirm the use of mean alpha level as a measure of resting-state arousal under eyes-closed and eyes-open conditions. The focal nature of EEG effects in the other bands suggests that these reflect cortical processing of visual input, producing differences in activation between eyes-closed and eyes-open resting conditions, rather than just the simple increase in arousal level shown in alpha. SIGNIFICANCE: This study demonstrates that the eyes-closed and eyes-open conditions provide EEG measures differing in topography as well as power levels. These differences should be recognised when evaluating EEG research, and considered when choosing eyes-open or eyes-closed baseline conditions for different paradigms. PMID- 17911043 TI - Statistical MUNE: a comparison of two methods of setting recording windows in healthy subjects and ALS patients. AB - OBJECTIVE: To address the issue as to how best to perform statistical MUNE, we applied two different approaches and compared results in healthy subjects and ALS patients. METHODS: Twelve normal subjects (women 8, mean age 52years) and 11 ALS patients (women 4, mean age 54years) underwent two consecutive MUNE studies, which differed in terms of setting and modifying the recording window. These are referred to as the 'expansion' and 'narrowing' methods, respectively. Size weighted average (Av) SMUP and MUNE values were obtained using the two methods, and compared in control and patient groups. RESULTS: Expansion method-derived Av SMUP sizes and MUNE values differed only slightly from those obtained using the narrowing method in healthy subjects, whereas the narrowing method resulted in significantly larger Av SMUP sizes and smaller MUNE values than the expansion method in ALS patients (Wilcoxon signed ranks test, p=0.003). The sizes of tested areas (mean+/-SD) were significantly larger for the narrowing method than the expansion method in both subject groups with much greater difference in ALS patients; 9.6+/-3.1% vs. 7.9+/-1.7% in healthy subjects and 16.1+/-5.1% vs. 11.2+/-3.0% in ALS patients (Student t-test, p<0.01). CONCLUSIONS: The present study shows, unlike that found in normal subjects, that the results of statistical MUNE in ALS patients are heavily dependent on the approach used to set and modify recording windows. SIGNIFICANCE: The expansion method using a 10% sized window is likely to suffer from systemic errors due to the ceiling effect and the sampling of artifactually small motor units in ALS patients. The authors recommend that the narrowing method be considered as an alternative that avoids these problems. PMID- 17911045 TI - Developing common methods for evaluating health information exchange. PMID- 17911044 TI - The role of CCL22/macrophage-derived chemokine in allergic rhinitis. AB - Dendritic cells (DCs) are considered to be the most powerful antigen-presenting cells (APCs). DCs are thought to be associated with Th1 or Th2 polarization and with polarization-induced disease such as atopic dermatitis, asthma and allergic rhinitis, but its mechanism is not well known. In this study, we analyzed the mRNA expression of DCs between birch pollen allergic rhinitis and healthy controls by using cDNA array. We found that the expressions of CCL22/macrophage derived chemokine (MDC) differed significantly. We also revealed that CCL22/MDC production was higher in patients than in healthy donors. By chemotaxis assay, CCL22/MDC can enhance the migration of patient's T cells rather than those of healthy controls. Surface marker analysis of migrated cells revealed that the most of migrated cells expressed CCR4, which were considered to be Th2 cells. Furthermore, CD1a(+) CD83(+) cells located in the nasal mucosa expressed CCL22/MDC in vivo. To the best of our knowledge, this is the first report clearly indicating the role of CCL22/MDC in allergic rhinitis. PMID- 17911046 TI - Identification of genes expressed in the mouse limb using a novel ZPA microarray approach. AB - One well-studied signaling center in the developing vertebrate limb, the zone of polarizing activity (ZPA), produces the morphogen sonic hedgehog that is necessary for normal growth and pattern formation. To identify additional factors expressed in the ZPA of the mouse limb bud, the Shhgfpcre allele was used to purify ZPA cells using fluorescence-activated cell sorting. Microarray technology was then used to identify genes whose expression was elevated in the ZPA compared to the rest of the limb. In situ hybridization confirmed the expression of two known transcription factors, Hlxb9 and Tcfap2b, an uncharacterized EST, and a transmembrane protein of unknown function in domains overlapping the ZPA. The expression of two other genes was confirmed by rtPCR. The methods described in this report will be applicable for identifying genes enriched in Shh-expressing cells throughout development. PMID- 17911047 TI - Heterospecific alarm call recognition in a non-vocal reptile. AB - The ability to recognize and respond to the alarm calls of heterospecifics has previously been described only in species with vocal communication. Here we provide evidence that a non-vocal reptile, the Galapagos marine iguana (Amblyrhynchus cristatus), can eavesdrop on the alarm call of the Galapagos mockingbird (Nesomimus parvulus) and respond with anti-predator behaviour. Eavesdropping on complex heterospecific communications demonstrates a remarkable degree of auditory discrimination in a non-vocal species. PMID- 17911048 TI - Context-dependent genetic benefits of polyandry in a marine hermaphrodite. AB - Numerous studies emphasize the potential indirect (genetic) benefits of polyandry in animals with resource-free mating systems. In this paper, we examine the potential for these benefits to fuel sexual selection and polyandry in the hermaphroditic ascidian Pyura stolonifera. Individuals were designated either sire (sperm producers) or dam (egg producers) at random and crossed in a North Carolina II breeding design to produce both paternal and maternal half siblings for our quantitative genetic analysis. We then partitioned the phenotypic variance in fertilization and hatching rates into additive and non-additive variance components. We found significant additive variance attributable to sire and dam effects at fertilization and hatching, suggesting the potential for selection to favour individuals carrying intrinsically 'good genes' for these traits. In separate analyses involving monandrous and polyandrous clutches, we found that both traits were elevated under polyandry, but the difference in hatching rates was due entirely to the difference in fertilization rates between treatments. When the hatching rates were standardized to account for variance at fertilization, there was no overall net benefit of polyandry for this trait. Despite this, we found that hatching success declined with increasing embryo densities, and that the slope of this decline was significantly greater in monandrous than polyandrous clutches. Hence, selection on embryo viability may still favour polyandry under restricted environmental conditions. Nevertheless, our results caution against interpreting elevated hatching success as an indirect genetic benefit of polyandry when variance in fertilization is not controlled. PMID- 17911049 TI - Receiver bias for exaggerated signals in honeybees and its implications for the evolution of floral displays. AB - Mechanistic models of animal signals posit the occurrence of biases on the part of receivers that could be potentially exploited by signallers. Such biases are most obvious when animals are confronted with exaggerated versions of signals they normally encounter. Signalling systems operating in plant-pollinator interactions are among the most highly coevolved, with plants using a variety of floral signals to attract pollinators. A number of observations suggest that pollinators preferentially visit larger floral displays although the benefit of this to either the plant or the pollinator is not always clear. We use a standard dual-choice experimental protocol to show that honeybees display a receiver bias for exaggerated size and colour contrast--two important components of floral signals--even when such signals do not indicate quality. We discuss the implications of this receiver bias for the evolution of floral displays and its possible exploitation by invading alien plants. PMID- 17911050 TI - Effects of metabolic rate on protein evolution. AB - Since the modern evolutionary synthesis was first proposed early in the twentieth century, attention has focused on assessing the relative contribution of mutation versus natural selection on protein evolution. Here we test a model that yields general quantitative predictions on rates of protein evolution by combining principles of individual energetics with Kimura's neutral theory. The model successfully predicts much of the heterogeneity in rates of protein evolution for diverse eukaryotes (i.e. fishes, amphibians, reptiles, birds, mammals) from different thermal environments. Data also show that the ratio of non-synonymous to synonymous nucleotide substitution is independent of body size, and thus presumably of effective population size. These findings indicate that rates of protein evolution are largely controlled by mutation rates, which in turn are strongly influenced by individual metabolic rate. PMID- 17911051 TI - Steroid hormones in bluegill, a species with male alternative reproductive tactics including female mimicry. AB - The proximate mechanisms underlying the evolution and maintenance of within-sex variation in mating behaviour are still poorly understood. Species characterized by alternative reproductive tactics provide ideal opportunities to investigate such mechanisms. Bluegill (Lepomis macrochirus) are noteworthy in this regard because they exhibit two distinct cuckolder (parasitic) morphs (called sneaker and satellite) in addition to the parental males that court females. Here we confirm previous findings that spawning cuckolder and parental males have significantly different levels of testosterone and 11-ketotestosterone. We also report, for the first time, that oestradiol and cortisol levels are higher in cuckolders than in parental males. The two cuckolder morphs did not differ in average levels of any of the four hormones. However, among satellite males which mimic females in appearance and behaviour, there was a strong negative relationship between oestradiol levels and body length, a surrogate for age. This finding suggests that for satellite males, oestradiol dependency of mating behaviour decreases with increasing mating experience. Although such decreased hormone dependence of mating behaviour has been reported in other taxa, our data represent the first suggestion of the relationship in fishes. PMID- 17911053 TI - Intact automatic imitation of human and robot actions in autism spectrum disorders. AB - The existence of a specialized imitation module in humans is hotly debated. Studies suggesting a specific imitation impairment in individuals with autism spectrum disorders (ASD) support a modular view. However, the voluntary imitation tasks used in these studies (which require socio-cognitive abilities in addition to imitation for successful performance) cannot support claims of a specific impairment. Accordingly, an automatic imitation paradigm (a 'cleaner' measure of imitative ability) was used to assess the imitative ability of 16 adults with ASD and 16 non-autistic matched control participants. Participants performed a prespecified hand action in response to observed hand actions performed either by a human or a robotic hand. On compatible trials the stimulus and response actions matched, while on incompatible trials the two actions did not match. Replicating previous findings, the Control group showed an automatic imitation effect: responses on compatible trials were faster than those on incompatible trials. This effect was greater when responses were made to human than to robotic actions ('animacy bias'). The ASD group also showed an automatic imitation effect and a larger animacy bias than the Control group. We discuss these findings with reference to the literature on imitation in ASD and theories of imitation. PMID- 17911054 TI - Functional significance of an unusual chela dimorphism in a marine decapod: specialization as a weapon? AB - The squat lobster Munida rugosa has an unusual chela dimorphism exhibited mainly by large males. Some individuals have 'arched' chelae in which there is a gap between the dactylus and the pollex when closed, and others have a 'straight' morphology in which the dactylus and pollex oppose along most of their length. Geometric morphometric analysis indicated that, compared with males, the arched morphology does not develop fully in females, so further investigation was confined to males. In males, the distal part of the chela was similar in both the forms and seemed to be adapted to hold and shred prey items. Both morphologies had a major cylindrical tooth on the inner proximal part of the dactylus, but the arched morphology had a higher and wider propodus, a greater major tooth-pollex distance and a greater force generation than the straight morphology. The findings suggest that the arched chela morphology in M. rugosa is a sexually selected trait adapted to inflict puncture wounds on opponents during agonistic interactions. The arched morphology, therefore, appears to have evolved in males by means of sexual selection because it enhanced the function of the chela as a weapon, while retaining functionality for feeding. PMID- 17911052 TI - Stochastic fluctuations in epidemics on networks. AB - The effects of demographic stochasticity on the long-term behaviour of endemic infectious diseases have been considered for long as a necessary addition to an underlying deterministic theory. The latter would explain the regular behaviour of recurrent epidemics and the former the superimposed noise of observed incidence patterns. Recently, a stochastic theory based on a mechanism of resonance with internal noise has shifted the role of stochasticity closer to the centre stage, by showing that the major dynamic patterns found in the incidence data can be explained as resonant fluctuations, whose behaviour is largely independent of the amplitude of seasonal forcing, and by contrast very sensitive to the basic epidemiological parameters. Here we elaborate on that approach, by adding an ingredient which is missing in standard epidemic models, the 'mixing network' through which infection may propagate. We find that spatial correlations have a major effect on the enhancement of the amplitude and the coherence of the resonant stochastic fluctuations, providing the ordered patterns of recurrent epidemics, whose period may differ significantly from that of the small oscillations around the deterministic equilibrium. We also show that the inclusion of a more realistic, time-correlated recovery profile instead of exponentially distributed infectious periods may, even in the random-mixing limit, contribute to the same effect. PMID- 17911055 TI - Fighting for shells: how private information about resource value changes hermit crab pre-fight displays and escalated fight behaviour. AB - Pre-fight displays typically provide honest, but sometimes dishonest, information about resource holding potential and may be influenced by assessment of resource value and hence motivation to acquire the resource. These assessments of potential costs and benefits are also predicted to influence escalated fight behaviour. This is examined in shell exchange contests of hermit crabs in which we establish an information asymmetry about a particularly poor quality shell. The poor shell was created by gluing sand to the interior whereas control shells lacked sand and the low value of the poor shell could not be accurately assessed by the opponent. Crabs in the poor shell showed changes in the use of pre-fight displays, apparently to increase the chances of swapping shells. When the fights escalated, crabs in poor shells fought harder if they took the role of attacker but gave up quickly if in the defender role. These tactics appear to be adaptive but do not result in a major shift in the roles taken or outcome. We thus link resource assessment with pre-fight displays, the roles taken, tactics used during escalation and the outcome of these contests. PMID- 17911056 TI - Device occlusion of Fontan fenestration--an economical alternative. AB - Fenestrating the Fontan circuit during total cavopulmonary anastomosis is commonly performed to reduce postoperative mortality and morbidity. The resulting systemic desaturation may progressively increase leading to symptoms. We report the case of a symptomatic eight year old, whose Fontan circuit fenestration was closed using a patent ductus arteriosus occluder. The marked improvement in the patient's clinical status immediately and after 3 months confirms this device to be a safe, and economically better alternative to the atrial septal defect occluder. PMID- 17911057 TI - Chylothorax following cardiac surgery caused by unusual lymphatic anatomy. AB - Chylothorax due to injury to the thoracic duct and lymphatic channels during left internal thoracic artery harvest is well described. However, high volume leakage of chyle due to disruption of aberrant thymic collateral lymphatic channel in the anterior mediastinum has not been described previously. We describe such a case which was managed by early surgical exploration, ligation of the aberrant duct, and insertion of a pleuro-peritoneal shunt. PMID- 17911058 TI - Primary hydatid cyst of the posterior mediastinum. AB - Hydatid cystic disease is still a health problem in Morocco. Mediastinal localization is very rare. We report the case of a 24-year-old man who presented with right chest pain and dyspnea. Chest X-Ray and computed tomography showed a mediastinal mass. The diagnosis was confirmed perioperatively by macroscopic and histologic investigations. Resection of the cyst was performed, and a small part intimately adherent to the inferior vena cava and oesophagus was left in place. There were no complications or recurrence. PMID- 17911059 TI - Two surgical cases of acute aortic dissection in pregnancy with marfan syndrome. AB - We describe 2 surgical cases of acute aortic type A dissection during pregnancy in women with Marfan syndrome. Both of them underwent emergency aortic root replacement under deep hypothermia; one patient was in her 21(st) week of pregnancy and the other was treated 1 day after a normal delivery. The patients experienced fair postoperative courses, but intrauterine fetus death was confirmed in the first case. PMID- 17911060 TI - Changing pattern of thoracic diseases in Korea over the last 25 years. PMID- 17911061 TI - Left ventricular outflow tract after Kawashima intraventricular rerouting. AB - Left ventricular outflow tract obstruction is a concern in the long term after Kawashima intraventricular rerouting for Taussig-Bing anomaly. This study assessed the late results and left ventricular outflow tract dimensions after intraventricular rerouting for Taussig-Bing anomaly in 6 children. Age at operation ranged from 8 months to 7 years. The shortest distance from the tricuspid valve to the pulmonary valve ranged from 8 to 18 mm, which was 79%-139% of the normal aortic annulus diameter. All patients survived the operation, but one died of chronic heart failure 4 months after repair. Three patients had no left ventricular outflow tract obstruction 15 to 31 years after repair, and 2 had significant obstruction at 9 and 14 years. One patient underwent successful revision of the intraventricular baffle; the distance between the tricuspid and pulmonary valves was 16 mm (108% of the normal aortic annulus) although it had been 8 mm (79%) at the initial repair. Late results of intraventricular rerouting were considered favorable. PMID- 17911062 TI - Thymectomy for myasthenia gravis: prognostic factors in 70 patients. AB - Thymectomy has become increasingly accepted as an efficacious procedure for myasthenia gravis, with high rates of complete clinical remission. Predictors of the response to thymectomy for myasthenia gravis vary in the literature. We retrospectively reviewed the clinical records of 70 patients (63% female; mean age, 38 years) diagnosed with myasthenia gravis from August 1993 to August 2004, to determine the factors predicting outcome. Complications occurred in 20%, but there was no hospital mortality. Complete clinical remission was obtained postoperatively in 47%. Our results indicate that patients with less than 1 year's duration of disease have a better prognosis, and Osserman stages I, IIa, and IIb are also associated with higher clinical remission rates. Female patients have a better prognosis than males, and the younger the patient the better the outcome. Thymectomy is indicated for myasthenia gravis as early as possible in the course of the disease. PMID- 17911063 TI - Continuous hemodiafiltration during cardiopulmonary bypass in infants. AB - The homologous blood prime in cardiopulmonary bypass circuits contributes a significant electrolyte and metabolite load in small infants. The efficacy of hemofiltration and continuous hemodiafiltration of the blood prime in preventing metabolic disturbances in small infants was compared in two groups of 60 patients each. Blood pH, base excess, and calcium concentrations decreased during cardiopulmonary bypass in the hemofiltration group. The acid-base balance was well preserved during cardiopulmonary bypass by continuous hemodiafiltration. This therapeutic strategy may confer an advantage in maintaining more physiological conditions during cardiopulmonary bypass in small infants. PMID- 17911064 TI - Total arterial revascularization and concomitant aortic valve replacement. AB - The safety of total arterial revascularization with a left internal thoracic artery-radial artery T-graft was evaluated in patients with at least two-vessel coronary artery disease and aortic valve stenosis requiring concomitant aortic valve replacement. From June 2001 to January 2005, 18 patients underwent aortic valve replacement and total arterial revascularization, while 101 had aortic valve replacement and conventional grafting. By matching age, sex, left ventricular ejection fraction, and number of distal anastomoses, 1:2 matched groups were generated: 15 patients with a left internal thoracic-radial artery T graft, and 30 with left internal thoracic artery and additional vein grafts. Aortic cross clamp and cardiopulmonary bypass times were similar in both groups. There were no significant differences in postoperative data between the groups. Early mortality was 0% in the T-graft group and 2% in those with conventional grafts. Follow-up ranged from 2 to 50 months. Event-free survival was 100% in the T-graft group and 90% in the conventional graft group. Total arterial grafting with a left internal thoracic-radial artery T-graft can be performed in selected patients with aortic valve stenosis requiring simultaneous aortic valve replacement. PMID- 17911065 TI - Results of isolated valve replacement in hemodialysis patients. AB - Frequent bleeding complications and poor long-term results have been reported after valve replacement in hemodialysis patients. We use mainly bileaflet mechanical valves with low-dose warfarin therapy (target international normalized ratio, 1.8-2.0) in such cases. Data of 27 hemodialysis patients undergoing isolated valve replacement from 1993 to 2002 were retrospectively analyzed. Bileaflet mechanical valves were selected in 23 patients and bioprostheses in 4. Those with mechanical valves were treated with mild anticoagulation therapy. There were 3 (11.1%) early deaths due to ischemic colitis, interstitial pneumonia, and ventricular arrhythmia. There were 3 late deaths and 5 bleeding complications during follow-up. The overall survival rate was 85.2% at 3 years and 72.9% at 5 years. The survival rate of patients with mechanical valves was 82.6% at 3 years and 76.7% at 5 years. One patient with a bioprosthesis experienced structural valvular deterioration after 3 years. The results demonstrate an acceptable long-term outcome. A bileaflet mechanical valve managed with mild anticoagulation therapy is a reasonable strategy for hemodialysis patients. PMID- 17911066 TI - Coronary artery bypass surgery with on-pump beating-heart technique. AB - To evaluate the results of on-pump beating-heart coronary bypass grafting, a retrospective study was carried out on 106 patients who had this procedure between 2003 and 2006. There were 87 (82%) men and 19 (18%) women, with a mean age of 60.53 +/- 11.97 years. Five (5%) patients had unstable angina, 10 (9%) had a recent myocardial infarction, and 16 (15%) had congestive heart failure. The mean ejection fraction was 40.38% +/- 11.46%. The mean cardiopulmonary bypass time was 66.81 +/- 31.14 min. The median number of grafts per patient was 3. The median intensive care unit stay was 47 hours, and hospital stay was 7 days. There were 4 (3.8%) deaths. The mean Parsonnet score was 12.75 +/- 11.27 and the logistic EuroSCORE was 7.06 +/- 8.62. This study shows that the on-pump beating heart technique is a safe and convenient method for coronary artery bypass grafting. PMID- 17911067 TI - Results of coronary bypass and valve operations for mitral valve regurgitation. AB - Combined coronary bypass grafting and valve procedures for mitral valve regurgitation result in poor outcomes, but the impact of the etiology of valve regurgitation on operative and long-term outcomes is not well defined. A retrospective analysis of 468 patients who had combined coronary bypass grafting and valve operations for mitral regurgitation showed that 78% had valve repairs and 22% had replacements for ischemic (45%) or degenerative (55%) disease. Predictors of operative mortality were ischemic mitral regurgitation, failure to use the internal mammary artery for grafting, severe coronary disease, acute myocardial infarction, low ejection fraction, advanced heart failure, emergency operation, and mitral valve replacement. The 5-year survival rates for propensity matched patients with ischemic or degenerative disease were similar (66%). Low ejection fraction (< 35%), advanced age (> 67 years), valve replacement surgery, residual mitral regurgitation, and severe coronary artery disease were predictors of poor long-term outcome. Although the operative outcomes of ischemic mitral regurgitation were poor compared to those of degenerative disease, the long-term survival was similar in both groups of propensity-matched patients. Left ventricular remodeling, an optimal valve procedure without residual mitral regurgitation, and left ventricular function are more important determinants of long-term outcome than the etiology of valve regurgitation. PMID- 17911068 TI - Urokinase thrombolysis in acute-on-chronic vascular occlusion of lower limb. AB - Between January 2000 and June 2003, 70 patients (63 men and 7 women) with acute on-chronic lower limb vascular occlusion underwent thrombolysis with 1 million units of urokinase. Forty-eight patients had unilateral and 22 had bilateral involvement; 9 had gangrenous changes. The mean age was 47 +/- 9 years, 73% were smokers, and 13% had diabetes. The definition of a good response was either return of distal pulses or a warm limb and relief of pain at rest. Fifty-three (76%) patients (including 5 with gangrenous changes) had symptomatic improvement with thrombolysis (group 1), and 17 (24%) did not respond (group 2). All 70 patients (including 7 who later required amputation) underwent surgical intervention with a polytetrafluoroethylene interposition graft or arterioplasty. A good response to surgical treatment was found in significantly more patients (49/53, 92%) in group 1 compared to group 2 (7/17, 41%). Claudication distance after 6 months of follow-up improved in a significantly higher percentage of patients in group 1 (85%) than group 2 (38%). Preoperative thrombolysis improved the outcome and predicted the result of arterial surgery. PMID- 17911069 TI - Long intraaortic balloon treatment time leads to more vascular complications. AB - Intraaortic balloon counterpulsation is an established and efficient therapy. Limb ischemia is the most common complication. The impact of treatment duration on balloon-related complications was analyzed retrospectively in 135 patients who underwent balloon counterpulsation between 1998 and 2004. Thirty high-risk coronary patients required preoperative intraaortic balloon therapy, 41 were in preoperative cardiogenic shock, and 64 needed support for difficulties in weaning from cardiopulmonary bypass. No balloon-related mortality occurred. The overall balloon-related complication rate was 20/135 (14.8%); 18 had limb ischemia, of whom 6 (4.4%) required vascular interventions. Intraaortic balloon treatment time was significantly longer in patients who developed limb ischemia (99.8 +/- 54.1 h) compared to those who did not (34.4 +/- 30.4 h). Preoperative therapy had short treatment times and few complications. Intraaortic balloon pumping provides effective circulatory support with a low complication rate. A clear relationship was established between duration of treatment and balloon-related complications. Independent risk factors for balloon-related complications were long treatment time, acute myocardial infarction, age over 65 years, and ejection fraction less than 0.30. PMID- 17911070 TI - Stentless valves treated by the L-hydro process in the aortic position in sheep. AB - Calcification of glutaraldehyde-treated bioprosthetic heart valves is a major cause of long-term failure. We studied porcine aortic valves treated by the L Hydro process and implanted into 14 juvenile sheep (group 1). Another 10 sheep were implanted with glutaraldehyde-treated porcine bioprostheses (group 2). The animals were sacrificed after 150 days and the explanted valves were analyzed for calcification. Hemodynamic measurements by echocardiography and angiography were carried out prior to sacrifice. Macroscopic analysis showed calcification and loss of mobility of the leaflets in all group 2 implants and in one group 1 implant. Light microscopy showed foci of calcification in all group 2 implants and in 3 valves from group 1. A significant reduction in the level of calcification was found in porcine bioprostheses treated by the L-Hydro process and implanted into the juvenile sheep model. PMID- 17911071 TI - Repair of anomalous origin of right coronary artery from the pulmonary artery. AB - Anomalous origin of the right coronary artery from the pulmonary artery is a rare cardiac malformation. Between July 2002 and July 2005, we operated on 4 patients with this defect. There were 2 males and 2 females, aged from 18 months to 42 years. Three patients underwent direct re-implantation of the right coronary artery into the aorta, and one had an intrapulmonary tunnel repair (intrapulmonary artery baffle with an autologous pericardial patch for tunneling to the anomalous right coronary ostium). Cardiopulmonary bypass was used in 2 patients, and an off-pump technique in the other 2. One patient had an atrial septal defect that was closed with an Amplatzer septal occluder through the right atrium under transesophageal echocardiography, without cardiopulmonary bypass. All patients survived and recovered uneventfully. Follow-up ranged from 3 to 39 months (mean, 17 months). All patients were doing well and free from symptoms, with normal exercise tolerance. Surgical correction of anomalous right coronary artery shows good early and midterm results. PMID- 17911072 TI - Surgical experience with cardiac echinococcosis. AB - Cardiac echinococcosis is a rare affliction of the heart encountered in areas where the tapeworm Echinococcus is endemic. During the 15 years to September 2005, 19 consecutive patients (11 female; mean age, 25.6 +/- 12.8 years) with cardiac echinococcosis were operated on at Damascus University. Diagnosis was mainly established by 2D-echocardiography. Twelve patients also had extracardiac hydatidosis. All patients were operated on using cardiopulmonary bypass and cardioplegia. Intact cysts were generally enucleated, ruptured cysts were exteriorized, and interventricular septal cysts were managed by controlled puncture and aspiration. There was no operative mortality. All patients were placed on mebendazole postoperatively for periods appropriate to the extent of their hydatidosis. There were 4 known late deaths due to systemic extension of the disease. Cardiac echinococcus is a serious disease that should be managed surgically without delay. Enucleation of intact cysts under cardiopulmonary bypass and cardiac arrest is our preferred method of excision. Anthelmintic medical treatment is a valuable and necessary adjunct to surgical management. PMID- 17911073 TI - Best site on right ventricle for open-chest biventricular pacing. AB - Cardiac resynchronization therapy is effective in patients with a low ejection fraction and left bundle branch block, but 20%-30% do not respond despite selection of the optimal site for pacing on the left ventricle. We investigated whether optimizing the site for placement of the pacing lead on the right ventricle could further improve left ventricular function during cardiac resynchronization in 19 patients (mean age, 63 +/- 5 years) undergoing coronary artery bypass with post-ischemic dilated myocardiopathy (ejection fraction, 25.8% +/- 2%) and left bundle branch block. The hemodynamic response to pacing was tested with the right ventricular lead positioned at the interventricular septum, atrioventricular junction, acute margin, and the pulmonary trunk. Biventricular stimulation improved left ventricular function. When the right ventricular lead was sited at the interventricular septum, a significant improvement in all hemodynamic parameters compared to the other sites was obtained. Biventricular pacing is important to optimize cardiac resynchronization. Although further studies are needed to confirm these findings, accurate lead placement is recommended for cardiac resynchronization therapy in patients with poor cardiac function and left bundle branch block. PMID- 17911074 TI - Juvenile rheumatic mitral stenosis with multiple ventricular septal defects. AB - Congenital heart disease is related to events occurring in the embryonal stage, while rheumatic heart disease is a sequela of immune-mediated damage following streptococcal infection. We report an unusual association of multiple ventricular septal defects and severe pulmonary arterial hypertension with rheumatic mitral stenosis in a 7-year-old girl. This case highlights the need for careful examination for coexisting rheumatic disease in late presentations of congenital heart disease. PMID- 17911075 TI - Atrial septal defect with ascites and pleural effusion. AB - A case of a lady referred for repair of an atrial septal defect is described. She presented with an insidious onset of recurrent ascites and pleural effusion. Cardiac catheterization showed constrictive physiology. The patient subsequently underwent surgical closure of the atrial septal defect and pericardiectomy. PMID- 17911076 TI - Congenital pulmonary vein stenosis with anomalous pulmonary venous connection. AB - An 11-month-old boy with congenital pulmonary vein stenosis, partial anomalous pulmonary venous connection, and ventricular septal defect is described. Angiocardiography demonstrated stenosis between the right upper pulmonary vein and high superior vena cava and obstruction of the right lower pulmonary vein. For pulmonary vein stenosis, we performed transverse sutured plasty for the right upper pulmonary vein, followed by right lower lobectomy. In some patients, combined management for pulmonary vein stenosis is effective. PMID- 17911077 TI - Benefits of a beta-blocker for intractable hemolysis due to paraprosthetic leakage. AB - We describe a 58-year-old man who was successfully treated with a beta-adrenergic receptor blocking agent for intractable hemolysis due to paraprosthetic leakage. After replacement of a mitral prosthetic valve with another mechanical valve, the patient suffered intractable intravascular hemolysis resulting from recurrent paraprosthetic leakage. With oral administration of a beta-adrenergic receptor blocker, betaxolol hydrochloride, for 3 months, the hemoglobin value increased from 9.7 g x dL(-1) to 12.4 g x dL(-1), although glutamic oxaloacetic transaminase and lactic dehydrogenase values remained elevated. PMID- 17911078 TI - Easy technique for mounting the Heartstring system into the sheath. AB - The Heartstring system was developed to avoid the use of side-biting aortic clamps, but the sealing system sometimes tears. To solve this problem, a string is wound around the coiled seal and pulled gently. PMID- 17911079 TI - Orthotopic heart transplantation through minimally invasive approach. AB - Minimal access approaches are a trend in cardiothoracic surgery. Gained experience in these minimally invasive techniques have allowed its application to more complicated procedures, such as heart transplantation. Both classic and bicaval techniques of cardiac transplant were performed through a partial lower sternotomy in 10 end-stage heart failure patients with no previous cardiac surgery. The procedure was considered safe with adequate exposure, minimal postoperative pain medication requirements, acceptable operative times, and good long-term outcome. PMID- 17911080 TI - Evaluating the quality of trials of hypothermic circulatory arrest aortic surgery. AB - The quality of level 1 evidence in reports on deep hypothermic circulatory arrest was assessed, and the confounding factors in surgical management and study design that can prevent meta-analysis formulation were determined. A systematic search of the literature was conducted using categorized nomenclature for randomized controlled trials in adult patients undergoing deep hypothermic circulatory arrest in the last 40 years. Twelve randomized controlled trials (2.3%) were found among 504 publications on deep hypothermic circulatory arrest listed on Medline from 1960; only 4 of them related to adults. One adequately powered study demonstrated reduced blood loss in deep hypothermic circulatory arrest using aprotinin. Three studies comparing retrograde and antegrade perfusion were underpowered. The median CONSORT score was 14 (range, 13-15). There were no consistent measures of similar outcomes (neuropsychometric, neurocognitive). No explanation was provided for the difference in reported ranges of neurological deficits in nonrandomized (5%-70%) and randomized (3%-9%) studies. Existing studies of deep hypothermic circulatory arrest are insufficient and inconsistent in the outcome measured, which explains the lack of a meta-analysis. Neurological injury remains high, and an appropriately powered study of interventions that can optimize cerebral perfusion is necessary. PMID- 17911081 TI - A right ventricle to aorta fistula caused by a fractured sternal wire. PMID- 17911082 TI - Subaortic valvular vegetation mimicking severe aortic valve stenosis. PMID- 17911087 TI - Proximal calciphylaxis treated with calcimimetic 'Cinacalcet'. AB - Calciphylaxis is a rare condition affecting patients suffering from end-stage renal failure, characterised by cutaneous ischaemia and necrosis. The management of calciphylaxis is challenging owing to the lack of optimal medical therapy, although parathyroidectomy has shown some benefit. We present a case of severe proximal calciphylaxis treated with a small dose of the calcimimetic 'Cinacalcet'. PMID- 17911086 TI - Characterization of the human skeletal muscle proteome by one-dimensional gel electrophoresis and HPLC-ESI-MS/MS. AB - Changes in protein abundance in skeletal muscle are central to a large number of metabolic and other disorders, including, and perhaps most commonly, insulin resistance. Proteomics analysis of human muscle is an important approach for gaining insight into the biochemical basis for normal and pathophysiological conditions. However, to date, the number of proteins identified by this approach has been limited, with 107 different proteins being the maximum reported so far. Using a combination of one-dimensional gel electrophoresis and high performance liquid chromatography electrospray ionization tandem mass spectrometry, we identified 954 different proteins in human vastus lateralis muscle obtained from three healthy, nonobese subjects. In addition to a large number of isoforms of contractile proteins, we detected all proteins involved in the major pathways of glucose and lipid metabolism in skeletal muscle. Mitochondrial proteins accounted for 22% of all proteins identified, including 55 subunits of the respiratory complexes I-V. Moreover, a number of enzymes involved in endocrine and metabolic signaling pathways as well as calcium homeostasis were identified. These results provide the most comprehensive characterization of the human skeletal muscle proteome to date. These data hold promise for future global assessment of quantitative changes in the muscle proteome of patients affected by disorders involving skeletal muscle. PMID- 17911085 TI - Periplasmic proteins of the extremophile Acidithiobacillus ferrooxidans: a high throughput proteomics analysis. AB - Acidithiobacillus ferrooxidans is a chemolithoautotrophic acidophile capable of obtaining energy by oxidizing ferrous iron or sulfur compounds such as metal sulfides. Some of the proteins involved in these oxidations have been described as forming part of the periplasm of this extremophile. The detailed study of the periplasmic components constitutes an important area to understand the physiology and environmental interactions of microorganisms. Proteomics analysis of the periplasmic fraction of A. ferrooxidans ATCC 23270 was performed by using high resolution linear ion trap-FT MS. We identified a total of 131 proteins in the periplasm of the microorganism grown in thiosulfate. When possible, functional categories were assigned to the proteins: 13.8% were transport and binding proteins, 14.6% were several kinds of cell envelope proteins, 10.8% were involved in energy metabolism, 10% were related to protein fate and folding, 10% were proteins with unknown functions, and 26.1% were proteins without homologues in databases. These last proteins are most likely characteristic of A. ferrooxidans and may have important roles yet to be assigned. The majority of the periplasmic proteins from A. ferrooxidans were very basic compared with those of neutrophilic microorganisms such as Escherichia coli, suggesting a special adaptation of the chemolithoautotrophic bacterium to its very acidic environment. The high throughput proteomics approach used here not only helps to understand the physiology of this extreme acidophile but also offers an important contribution to the functional annotation for the available genomes of biomining microorganisms such as A. ferrooxidans for which no efficient genetic systems are available to disrupt genes by procedures such as homologous recombination. PMID- 17911088 TI - Influence of renal involvement on peripheral blood mononuclear cell expression behaviour of tumour necrosis factor-alpha and interleukin-6 in type 2 diabetic patients. AB - BACKGROUND: Type 2 diabetes is associated with a high cardiovascular risk, which is even increased if renal damage is superimposed. Peripheral blood mononuclear cells (PBMCs) and pro-inflammatory cytokines are key factors linking type 2 diabetes and atherosclerosis. We investigated the influence of renal damage on serum, urinary and PBMCs expression behavior of TNF-alpha and IL-6 in these patients. METHODS: PBMCs were isolated by density gradient centrifugation (Ficoll Paque method) from fasting blood samples of 22 non-diabetic control subjects and 78 diabetic patients with normal renal function and different stages of diabetic nephropathy (18 with normoalbuminuria, 29 with microalbuminuria and 31 with macroalbuminuria). Expression levels of TNF-alpha and IL-6 were analyzed by real time quantitative RT-PCR. Serum and urinary TNF-alpha and IL-6 concentrations were measured by a solid-phase, chemiluminescent immunometric assay. RESULTS: The mean percent increases in the serum and urinary levels of TNF-alpha and IL-6 in diabetic patients with respect to control subjects were 176% (P < 0.0001), 250% (P < 0.0001), 114% (P < 0.0001) and 39.6% (P = 0.01), respectively. The mRNA expression level of TNF-alpha was higher by 68.8% (P < 0.001) and IL-6 mRNA levels were higher by 64.1% (P < 0.001) with respect to non-diabetic controls. TNF-alpha mRNA expression in patients with macroalbuminuria was higher by 84.8% with respect to subjects with normalbuminuria (P < 0.001) and by 29% with respect to individuals with microalbuminuria (P < 0.05). Likewise, microalbuminuric patients showed a 44.5% increase in TNF-alpha mRNA expression compared to subjects with normoalbuminuria (P < 0.05). Concerning IL-6, the mRNA expression levels of this cytokine was higher by 63.1% with respect to normoalbuminuric subjects (P < 0.01), and by 23.1% with respect to patients with microalbuminuria (P < 0.05). However, with respect to controls, diabetic patients with normoalbuminuria had similar serum TNF-alpha and urinary excretion of IL-6, without any differences in the mRNA expression levels of these cytokines in PBMCs. Partial correlation and multiple regression analysis using TNF-alpha and IL-6 mRNA levels as the dependent variables showed that urinary albumin excretion (UAE) was direct and independently associated with the expression profile of these pro-inflammatory cytokines in PBMCs. CONCLUSIONS: These data show for the first time the relationship between inflammatory activation of PBMCs (reflected by enhanced mRNA expression of TNF-alpha and IL-6) and renal involvement (reflected by increased UAE) in type 2 diabetic patients. These results provide potential insights for the increased inflammation, accelerated atherosclerosis and cardiovascular risk associated with nephropathy in type 2 diabetes. PMID- 17911089 TI - Gene expression analysis of kidneys from transgenic mice expressing fibroblast growth factor-23. AB - BACKGROUND: Fibroblast growth factor-23 (FGF23), a circulating protein produced in bone, causes decreased renal inorganic phosphate (Pi) reabsorption by reducing the expression of the sodium phosphate cotransporter type 2a (Npt2a). We have previously generated transgenic mice expressing human wild-type (WT) FGF23 under the control of the alpha1 (I) collagen promoter. METHODS: In this study, we performed a large-scale gene expression study of kidneys from FGF23 transgenic mice and WT littermates. Microarray expression data of key transcripts were verified by real-time RT-PCR analysis. RESULTS: Several genes that play a role in Pi regulation revealed decreased expression levels in the transgenic mice, such as Npt2a and Pdzk1, a scaffolding protein known to interact with Npt2a. Importantly, Klotho, a suggested FGF23 receptor cofactor, was the most significantly decreased transcript and alpha2-Na(+)/K(+)-ATPase (Atp1a2), a gene isoform of alpha1-Na(+)/K(+)-ATPase (Atp1a1) which has recently been shown to interact with Klotho and regulate calcium metabolism, was the most increased transcript. In contrast, other genes proposed to regulate Pi levels, such as secreted frizzled-related protein-4 (sFrp4) and Na(+)/H(+) exchanger regulatory factor-1 (Nherf1) revealed no changes. CONCLUSIONS: FGF23 transgenic mice display differentially expressed transcript levels of several genes essential in renal Pi regulation. These findings may lead to further understanding of how FGF23 mediates its actions on renal Pi regulation. PMID- 17911090 TI - A Comparison of GFR estimating formulae based upon s-cystatin C and s-creatinine and a combination of the two. AB - BACKGROUND: Current recommendations (KDIGO and NKF-K/DOQI) are that patients with chronic kidney diseases (CKD) should be classified in stages 1-5 based on GFR. A serum creatinine-based prediction equation (abbreviated MDRD formula) can be used to estimate GFR (eGFR). Cystatin C has been proposed as an alternative filtration marker to creatinine. We present validation of currently used formulae for eGFR based upon s-creatinine and s-cystatin C and we compare two different methods for the determination of cystatin C. METHODS: S-cystatin C and s-creatinine were measured in 644 patients referred for determination of GFR by plasma clearance of iohexol during the period 1 June 2004 to 31 December 2005. S-cystatin C was determined by turbidimetry using two different reagents (DAKO A/S and Gentian A/S). The 644 patients were divided into two groups. Group 1 was used to calculate own eGFR-formulae based on s-cystatin C (Orebro-cyst). Group 2 was used to validate the formulae. Three creatinine-based equations (Cockcroft-Gault, MDRD and Jelliffe) and seven cystatin C-based (Larsson, Hoek, Filler, leBricon, Grubb and Orebro-cyst DAKO, Gentian) were evaluated. Evaluation was done according to the recommendations by K/DOQI. RESULTS: In the test sample (group 2) mean GFR (iohexol clearance) was 50.4 ml/min/1.73 m(2) (range 12-150)-mean s-cystatin C (DAKO AS) was 1.63 mg/l and mean s-cystatin C (Gentian AS) 1.92 mg/l. The s cystatin C concentrations obtained by the Gentian method were approximately 10% lower than the DAKO method within the normal GFR range but were approximately 40% higher within the low GFR range. Bias for the creatinine-based equations was in the range -0.9 to 5.9 ml/min/1.73 m(2) and for the cystatin C-based equations in range -2.4 to 7.9 ml/min/ 1.73 m(2). Accuracy within 30% ranged from 68.6 to 80.4% and 54.0 to 82.9%, respectively. By combining both, an accuracy within 30% for 87.0% could be reached (MDRD/cystatin C by Gentian). Overall the patients were correctly classified for the different stages of CKD in 62.1-64.0% for the creatinine-based equations, 61.5-72.0% for the cystatin C-based equations and 70.2-73.9% for the combination. CONCLUSION: Estimating GFR using formulae based on s-creatinine or s-cystatin C alone was equally accurate according to the NKF K/DOQI guidelines. A formula that combines both provided a greater accuracy. If Cystatin C, which is clearly more expensive, is used, the choice of the cystatin C determination method and an adjusted prediction equation is essential. Use of the IDMS-traceable MDRD seems to yield the best cost-benefit ratio for routine practice. PMID- 17911091 TI - Tacrolimus as a steroid-sparing agent for adults with steroid-dependent minimal change nephrotic syndrome. AB - BACKGROUND: Treatment of adults with steroid-dependent minimal change nephrotic syndrome (SD-MCNS) can be a significant challenge. Cyclophosphamide (CYC) and cyclosporin (CYA) are often effective steroid-sparing agents. Tacrolimus (TAC) may be another treatment option. METHODS: This open, prospective cohort study enrolled Chinese adults with SD-MCNS. At the start of the study, we administered TAC or intravenous CYC together with prednisone (0.5 mg/kg/day), the dose of which was tapered off throughout the study. The TAC cohort received oral TAC (target trough blood level of 4-8 ng/ml) for 24 weeks and the CYC cohort received intravenous CYC (750 mg/m(2) body surface) once every 4 weeks for 24 weeks. RESULTS: Twenty-six patients met the criteria for enrollment (14 patients in the CYC group and 12 patients in the TAC group). One patient from each group discontinued treatment because of a drug-related side effect. Complete remission (CR) after the 24-week therapeutic period was 76.9% (10/13) in the CYC group and 90.9% (10/11) in the TAC group. The mean time required for CR in the TAC group was significantly less than in the CYC group (P = 0.031). Eight of 13 (61.5%) patients in the CYC group and 8 of 11 (72.7%) patients in the TAC group successfully stopped steroids and changed their status from steroid dependence. Sixty percent (6/10) of the CYC patients and 50% (5/10) of the TAC patients who achieved CR maintained remission during the follow-up period of 23.0 +/- 10.1 months. Four (40%) CYC patients and five (50%) TAC patients experienced relapses, and two CYC patients experienced frequent relapses. CONCLUSION: A 24-week course of TAC is a favorable steroid-sparing agent for treatment of Chinese adults with SD-MCNS. Therapy with TAC accompanied by a tapering dose of prednisolone appears to yield quicker remission than treatment with CYC together with prednisone. PMID- 17911092 TI - Sleep quality predicts quality of life and mortality risk in haemodialysis patients: results from the Dialysis Outcomes and Practice Patterns Study (DOPPS). AB - BACKGROUND: Poor sleep quality (SQ) affects many haemodialysis (HD) patients and could potentially predict their morbidity, mortality, quality of life (QOL) and patterns of medication use. METHODS: Data on SQ were collected from 11,351 patients in 308 dialysis units in seven countries in the Dialysis Outcomes and Practice Patterns Study (DOPPS) between 1996 and 2001 through a patient self reported SQ scale, ranging from 0 (worst) to 10 (best). A score of <6 reflected poor SQ. Sleep disturbance was also assessed by self-reported daytime sleepiness, feeling drained and nocturnal awakening. Logistic and multiple linear regression were used to assess predictors of SQ and associations with QOL. Cox regression examined associations with mortality. Analyses accounted for case-mix, facility clustering and country. RESULTS: Nearly half (49%) of patients experienced poor SQ. Mean SQ scores varied by country, ranging from 4.9 in Germany to 6.5 in Japan. Patients with poor SQ were more likely to be prescribed antihistamines, antidepressants, anti-inflammatories, narcotics, gastrointestinal (GI) medications, anti-asthmatics or hypnotics. Physical exercise at least once a week (vs < once a week) was associated with lower odds of poor SQ (AOR = 0.55-0.85, P < 0.05). Poorer SQ was associated with significantly lower mental and physical component summary (MCS/PCS) scores (MCS scores 1.9-13.2 points lower and PCS scores 1.5-7.7 points lower when SQ scores were <10 vs 10). The RR of mortality was 16% higher for HD patients with poor SQ. CONCLUSIONS: Poor SQ is common among HD patients in DOPPS countries and is independently associated with several QOL indices, medication use patterns and mortality. Assessment and management of SQ should be an important component of care. PMID- 17911093 TI - Enlarged kidneys and acute renal failure--why is a renal biopsy necessary for diagnosis and treatment? PMID- 17911094 TI - Urinary interleukin-18 is an acute kidney injury biomarker in critically ill children. AB - BACKGROUND: Urinary interleukin-18 (uIL-18) is an earlier acute kidney injury (AKI) biomarker than serum creatinine (SCr) in specific populations. In the present study, the relationship between uIL-18 and AKI was determined in a heterogeneous group of critically ill children. METHODS: We studied critically ill children to determine whether uIL-18 was an early predictor of AKI. SCr was determined daily for up to 14 days from mechanical ventilation initiation and up to four serial urine specimens were collected for the uIL-18 measurement. AKI was graded by paediatric modified risk, injury, failure, loss, end-stage kidney disease (pRIFLE) criteria. Day 0 was defined as the day of attaining pRIFLE AKI. RESULTS: One hundred thirty-seven children aged 6.5 +/- 6.4 years (53% male) were studied. The peak levels of IL-18 correlated with the severity of AKI by pRIFLE classification (P < 0.05). In non-septic AKI patients, uIL-18 rose to a level higher than control levels 2 days prior to a significant rise in SCr. Urinary IL 18 concentration from the first urine specimen was associated with AKI development within 48 h (odds ratio = 3.5, P < 0.05) independent of the paediatric risk of mortality (PRISM II) score. Urinary IL-18 concentration > or = 100 pg/ml had a specificity and negative predictive value of 81 and 83% to predict AKI development within 24 h. Urinary IL-18 > or = 200 pg/ml collected within 24 h of Day 0 had a specificity and positive predictive value of 93 and 88% respectively to predict the AKI duration > or = 48 h. Urinary IL-18 was associated with mortality (odds ratio = 1.29, P < 0.05), independent of the PRISM II score. CONCLUSIONS: Urinary IL-18 rises prior to SCr in non-septic critically ill children, predicts severity of AKI and is an independent predictor of mortality. PMID- 17911095 TI - Recurrent pulmonary oedema and severe hypertension after renal transplantation: other reasons than renal artery stenosis. PMID- 17911097 TI - The cyclophilin-like domain of Ran-binding protein-2 modulates selectively the activity of the ubiquitin-proteasome system and protein biogenesis. AB - The ubiquitin-proteasome system (UPS) plays a critical role in protein degradation. The 19S regulatory particle (RP) of the 26S proteasome mediates the recognition, deubiquitylation, unfolding, and channeling of ubiquitylated substrates to the 20S proteasome. Several subunits of the 19S RP interact with a growing number of factors. The cyclophilin-like domain (CLD) of Ran-binding protein-2 (RanBP2/Nup358) associates specifically with at least one subunit, S1, of the base subcomplex of the 19S RP, but the functional implications of this interaction on the UPS activity are elusive. This study shows the CLD of RanBP2 promotes selectively the accumulation of a subset of reporter substrates of the UPS, such as the ubiquitin (Ub)-fusion yellow fluorescent protein (YFP) degradation substrate, Ub(G76V)-YFP, and the N-end rule substrate, Ub-R-YFP. Conversely, the degradation of endoplasmic reticulum and misfolded proteins, and of those linked to UPS-independent degradation, is not affected by CLD. The selective effect of CLD on the UPS in vivo is independent of, and synergistic with, proteasome inhibitors, and CLD does not affect the intrinsic proteolytic activity of the 20S proteasome. The inhibitory activity of CLD on the UPS resides in a purported SUMO binding motif. We also found two RanBP2 substrates, RanGTPase activating protein and retinitis pigmentosa GTPase regulator interacting protein 1alpha1, whose steady-state levels are selectively modulated by CLD. Hence, the CLD of RanBP2 acts as a novel auxiliary modulator of the UPS activity; it may contribute to the molecular and subcellular compartmentation of the turnover of properly folded proteins and modulation of the expressivity of several neurological diseases. PMID- 17911096 TI - Functional SmpB-ribosome interactions require tmRNA. AB - Small protein B (SmpB) is a requisite component of the transfer messenger RNA (tmRNA)-mediated bacterial translational quality control system known as trans translation. The initial binding of tmRNA and its subsequent accommodation into the ribosomal A-site are activities intimately linked to SmpB protein function. From a mechanistic perspective, two key unanswered questions that require further investigation are: 1) what constitutes a stalled ribosome recognition complex and 2) does SmpB pre-bind ribosomes to recruit tmRNA. We have assessed, both in vivo and in vitro, the nature and stability of free SmpB interactions with stalled ribosomes and examined whether these interactions are functionally relevant. We present evidence to demonstrate that interaction of free SmpB with ribosomes is salt sensitive and significantly more labile than interaction of the SmpB.tmRNA complex with ribosomes. Upon dissociation of 70 S ribosomes SmpB partitions primarily with tmRNA rather than ribosomal subunits. This finding is consistent with biochemical and structural data demonstrating that tmRNA is the high affinity binding partner of SmpB. Moreover, we show that under normal physiological conditions roughly similar numbers of SmpB and tmRNA molecules are present in cells. Our investigations also reveal that upon induction of a nonstop mRNA, SmpB is enriched in stalled ribosome fractions only in the presence of tmRNA. Based on these findings, we conclude that SmpB does not pre-bind stalled ribosome and that functional SmpB-stalled ribosome interactions require tmRNA. We propose that a 1:1:1 complex of SmpB.tmRNA.EF-Tu(GTP) recognizes and binds a stalled ribosome to initiate trans-translation. PMID- 17911098 TI - A peptide-gated ion channel from the freshwater polyp Hydra. AB - Chemical transmitters are either low molecular weight molecules or neuropeptides. As a general rule, neuropeptides activate only slow metabotropic receptors. To date, only one exception to this rule is known, the FMRFamide-activated Na(+) channel (FaNaC) from snails. Until now FaNaC has been regarded as a curiosity, and it was not known whether peptide-gated ionotropic receptors are also present in other animal groups. Nervous systems first evolved in cnidarians, which extensively use neuropeptides. Here we report cloning from the freshwater cnidarian Hydra of a novel ion channel (Hydra sodium channel, HyNaC) that is directly gated by the neuropeptides Hydra-RFamides I and II and is related to FaNaC. The cells expressing HyNaC localize to the base of the tentacles, adjacent to the neurons producing the Hydra-RFamides, suggesting that the peptides are the natural ligands for this channel. Our results suggest that neuropeptides were already used for fast transmission in ancient nervous systems. PMID- 17911099 TI - Conformational variability of the glycine receptor M2 domain in response to activation by different agonists. AB - Models describing the structural changes mediating Cys loop receptor activation generally give little attention to the possibility that different agonists may promote activation via distinct M2 pore-lining domain structural rearrangements. We investigated this question by comparing the effects of different ligands on the conformation of the external portion of the homomeric alpha1 glycine receptor M2 domain. Conformational flexibility was assessed by tethering a rhodamine fluorophore to cysteines introduced at the 19' or 22' positions and monitoring fluorescence and current changes during channel activation. During glycine activation, fluorescence of the label attached to R19'C increased by approximately 20%, and the emission peak shifted to lower wavelengths, consistent with a more hydrophobic fluorophore environment. In contrast, ivermectin activated the receptors without producing a fluorescence change. Although taurine and beta-alanine were weak partial agonists at the alpha1R19'C glycine receptor, they induced large fluorescence changes. Propofol, which drastically enhanced these currents, did not induce a glycine-like blue shift in the spectral emission peak. The inhibitors strychnine and picrotoxin elicited fluorescence and current changes as expected for a competitive antagonist and an open channel blocker, respectively. Glycine and taurine (or beta-alanine) also produced an increase and a decrease, respectively, in the fluorescence of a label attached to the nearby L22'C residue. Thus, results from two separate labeled residues support the conclusion that the glycine receptor M2 domain responds with distinct conformational changes to activation by different agonists. PMID- 17911100 TI - Metal-catalyzed oxidation of the Werner syndrome protein causes loss of catalytic activities and impaired protein-protein interactions. AB - Metal-catalyzed oxidation reactions target amino acids in the metal binding pocket of proteins. Such oxidation reactions generally result in either preferential degradation of the protein or accumulation of a catalytically inactive pool of protein with age. Consistently, levels of oxidized proteins have been shown to increase with age. The segmental, progeroid disorder Werner syndrome results from loss of the Werner syndrome protein (WRN). WRN is a member of the RecQ family of DNA helicases and possesses exonuclease and ATP-dependent helicase activities. Furthermore, each of the helicase and exonuclease domains of WRN contains a metal binding pocket. In this report we examined for metal catalyzed oxidation of WRN in the presence of iron or copper. We found that WRN was oxidized in vitro by iron but not by copper. Iron-mediated oxidation resulted in the inhibition of both WRN helicase and exonuclease activities. Oxidation of WRN also inhibited binding to several known protein partners. In addition, we did not observe degradation of oxidized WRN by the 20 S proteasome in vitro. Finally, exposure of cells to hydrogen peroxide resulted in oxidation of WRN in vivo. Therefore, our results demonstrate that WRN undergoes metal-catalyzed oxidation in the presence of iron, and iron-mediated oxidation of WRN likely results in the accumulation of a catalytically inactive form of the protein, which may contribute to age-related phenotypes. PMID- 17911101 TI - The C-terminal extension of the beta7 subunit and activator complexes stabilize nascent 20 S proteasomes and promote their maturation. AB - The eukaryotic 20 S proteasome is formed by dimerization of two precursor complexes containing the maturation factor Ump1. Beta7/Pre4 is the only one of the 14 subunits forming the 20 S proteasome that is absent from these precursor complexes in Saccharomyces cerevisiae. Increased expression of Pre4 leads to a reduction in the level of precursor complex, indicating that Pre4 incorporation into these complexes is rate-limiting for their dimerization. When we purified these precursor complexes, we observed co-purification of Blm10, a large protein known to attach to the alpha ring surface of proteasomes. In contrast to single mutants lacking either Blm10 or the C-terminal extension of Pre4, a mutant lacking both grew extremely poorly, accumulated very high levels of precursor complexes, and was impaired in beta subunit maturation. The effect of blm10Delta on proteasome biogenesis is modest, apparently because the 19 S regulatory particle is capable of substituting for Blm10, as long as precursor complex dimers are stabilized by the Pre4 C terminus. We found that a mutation (sen3/rpn2) affecting the Rpn2 subunit inhibits attachment of the 19 S activator to the 20 S particle or its precursors. Although the sen3 mutation alone had no apparent effect on precursor complex dimerization and active site maturation, the sen3 blm10 double mutant was impaired in these processes. Together these data demonstrate that Blm10 and the 19 S activator have a partially redundant function in stabilizing nascent 20 S proteasomes and in promoting their activation. PMID- 17911103 TI - Quantification and modeling of tripartite CD2-, CD58FC chimera (alefacept)-, and CD16-mediated cell adhesion. AB - Alefacept is a chimeric protein combining CD58 immunoglobulin-like domain 1 with human IgG1 Fc. Alefacept mediates adhesion by bridging CD2 on T cells to activating Fc receptors on effector cells, but the equilibrium binding parameters have not been determined. Alefacept mediated T cell killing by NK cells and adhesion between CD2- and CD16-expressing cells at an optimum concentration of 100 nM. We introduce novel measurements with supported planer bilayers, from which key two-dimensional and three-dimensional parameters can be determined by data fitting. Alefacept competitively inhibited cell bilayer adhesion mediated by the CD2-CD58 interaction. Alefacept mediated maximal adhesion of CD2(+) T cells to CD16B, an Fc receptor, in planar bilayers at 500 nM. A mechanistic model for alefacept-mediated cell-bilayer adhesion allowed fitting of the data and determination of two-dimensional binding parameters. These included the density of bonds in the adhesion area, which grew to maintain a consistent average bond density of 200 molecules/microm(2) and two-dimensional association constants of 3.1 and 630 microm(2) for bivalently and monovalently bound forms of alefacept, respectively. The maximum number of CD16 bound and the fit value of 4,350 CD2 per cell are much lower than the 40,000 CD2 per cell measured with anti-CD2 Fab. These results suggest that additional information is needed to correctly predict Alefacept-mediated bridge formation. PMID- 17911102 TI - Keratocan and lumican regulate neutrophil infiltration and corneal clarity in lipopolysaccharide-induced keratitis by direct interaction with CXCL1. AB - Keratocan and lumican are keratan-sulfate proteoglycans (KSPG), which have a critical role in maintaining corneal clarity. To determine whether these KSPGs have a role in corneal inflammation, we examined Kera(-/-) and Lum(-/-) mice in a model of lipopolysaccharide (LPS)-induced keratitis in which wild-type mice develop increased corneal thickness and haze due to neutrophil infiltration to the corneal stroma. Corneal thickness increases caused by LPS mice were significantly lower in Kera(-/-) and Lum(-/-) than wild-type mice. Further, LPS injected Lum(-/-) mice had elevated corneal haze levels compared with that of Kera(-/-) and wild-type. At 24 h post-injection, total enhanced green fluorescent protein-positive bone marrow-derived inflammatory cells in chimeric mice was significantly lower in Kera(-/-) mice and Lum(-/-) mice compared with wild-type mice. Neutrophil infiltration was inhibited in Kera(-/-) and Lum(-/-) mice at 6 and 24 h post-stimulation, with Lum(-/-) corneas having the most profound defect in neutrophil migration. Reconstitution of keratocan and lumican expression in corneas of Kera(-/-) and Lum(-/-) mice using adeno-keratocan and adeno-lumican viral vectors, respectively, resulted in normal neutrophil infiltration in response to LPS. Immunoprecipitation/Western blot analysis showed that lumican and keratocan core proteins bind the CXC chemokine KC during a corneal inflammatory response, indicating that corneal KSPGs mediate neutrophil recruitment to the cornea by regulating chemokine gradient formation. Together, these data support a significant role for lumican and keratocan in a corneal inflammatory response with respect to edema, corneal clarity, and cellular infiltration. PMID- 17911104 TI - Substitution of tyrosine 146 in the dI component of proton-translocating transhydrogenase leads to reversible dissociation of the active dimer into inactive monomers. AB - Transhydrogenase couples the redox reaction between NADH and NADP+ to proton translocation across a membrane. The protein has three components: dI binds NADH, dIII binds NADP+, and dII spans the membrane. Transhydrogenase is a "dimer" of two dI-dII-dIII "monomers"; x-ray structures suggested that the two catalytic sites alternate during turnover. Invariant Tyr146 in recombinant dI of Rhodospirillum rubrum transhydrogenase was substituted with Phe and Ala (proteins designated dI.Y146F and dI.Y146A, respectively). Analytical ultracentrifuge experiments and differential scanning calorimetry show that dI.Y146A more readily dissociates into monomers than wild-type dI. Analytical ultracentrifuge and Trp fluorescence experiments indicate that the dI.Y146A monomers bind NADH much more weakly than dimers. Wild-type dI and dI.Y146F reconstituted activity to dI depleted membranes with similar characteristics. However, dI.Y146A reconstituted activity in its dimeric form but not in its monomeric form, this despite monomers retaining their native fold and binding to the dI-depleted membranes. It is suggested that transhydrogenase reconstructed with monomers of dI.Y146A is catalytically compromised, at least partly as a consequence of the lowered affinity for NADH, and this results from lost interactions between the nucleotide binding site and the protein beta-hairpin upon dissociation of the dI dimer. The importance of these interactions and their coupling to dI domain rotation in the mechanism of action of transhydrogenase is emphasized. Two peaks in the 1H NMR spectrum of wild-type dI are broadened in dI.Y146A and are tentatively assigned to S-methyl groups of Met resonances in the beta-hairpin, consistent with the segmental mobility of this feature in the structure. PMID- 17911105 TI - Active gamma-secretase complexes contain only one of each component. AB - Gamma-secretase is an intramembrane aspartyl protease complex that cleaves type I integral membrane proteins, including the amyloid beta-protein precursor and the Notch receptor, and is composed of presenilin, Pen-2, nicastrin, and Aph-1. Although all four of these membrane proteins are essential for assembly and activity, the stoichiometry of the complex is unknown, with the number of presenilin molecules present being especially controversial. Here we analyze functional gamma-secretase complexes, isolated by immunoprecipitation from solubilized membrane fractions and able to produce amyloid beta-peptides and amyloid beta-protein precursor intracellular domain. We show that the active isolated protease contains only one presenilin per complex, which excludes certain models of the active site that require aspartate dyads formed between two presenilin molecules. We also quantified components in the isolated complexes by Western blot using protein standards and found that the amounts of Pen-2 and nicastrin were the same as that of presenilin. Moreover, we found that one Aph-1 was not co-immunoprecipitated with another in active complexes, evidence that Aph 1 is likewise present as a monomer. Taken together, these results demonstrate that the stoichiometry of gamma-components presenilin:Pen-2:nicastrin:Aph-1 is 1:1:1:1. PMID- 17911106 TI - Lysosomal dysfunction results in altered energy balance. AB - The mucopolysaccharidosis (MPS) type VII mouse was originally described as the adipose storage deficiency mouse because of its extreme lean phenotype of unknown etiology. Here, we show that adipose storage deficiency and lower leptin levels are common to five different lysosomal storage diseases (LSDs): MPSI, MPSIIIB, MPSVII, Niemann-Pick type A/B, and infantile neuronal ceroid lipofuscinosis. Elevated circulating pro-inflammatory proteins (VCAM1 and MCP1) were found in multiple LSDs. Multiple anti-inflammatory strategies (dexamethasone, MCP1 deficiency, M3 expression) failed to alter adiposity in LSD animals. All of the models had normal or greater caloric intake and lower to normal metabolic rate, fasting plasma glucose, non-esterified fatty acids, cholesterol, and triglycerides. Triglycerides were lower in the livers of MPSI mice, and the trend was lower in the muscle. Lipid absorption and processing in MPSI mice were indistinguishable from those in normal mice following oral gavage of olive oil. The increased lean mass of MPSI and MPSIIIB mice suggests a shift in adipose triglycerides to lysosomal storage. In agreement, MPSI livers had a similar total caloric content but reduced caloric density, indicating a shift in energy from lipids to proteins/carbohydrates (lysosomal storage). Enzyme replacement therapy normalized the caloric density within 48 h without reducing total caloric content. This was due to an increase in lipids. Recycling of stored material is likely reduced or nonexistent. Therefore, to maintain homeostasis, energy is likely diverted to synthesis at the expense of typical energy storage depots. Thus, these diseases will serve as important tools in studying the role of lysosome function in metabolism and obesity. PMID- 17911107 TI - Substitutions of potentially phosphorylatable serine residues of Bax reveal how they may regulate its interaction with mitochondria. AB - During apoptosis, the pro-apoptotic protein Bax relocalizes from the cytosol to the mitochondrial outer membrane. This relocalization is associated to major conformational changes, namely at the N- and C-terminal ends of the protein. Substitution of residues located at critical positions within the protein potentially stimulates or inhibits this process. In the present study, we investigated the hypothesis that phosphorylation of serine residues might trigger these conformational changes, with a focus on Ser(163) and Ser(184), which have been shown to be phosphorylatable by protein kinases GSK3beta and Akt/PKB, respectively, and on Ser(60), which is located in a consensus target sequence for PKA. Substitutions of these serine residues by alanine or aspartate were done in wild type or previously characterized Bax mutants, and the capacity of the resulting proteins to interact with mitochondria and to release cytochrome c was assayed in yeast, which provides a tool to study the function of Bax, independently of the rest of the apoptotic network. We conclude that sequential phosphorylation of these serine residues might participate in the triggering of the different conformational changes associated with Bax activation during apoptosis. PMID- 17911108 TI - A charge reversal differentiates (p)ppGpp synthesis by monofunctional and bifunctional Rel proteins. AB - A major regulatory mechanism evolved by microorganisms to combat stress is the regulation mediated by (p)ppGpp (the stringent response molecule), synthesized and hydrolyzed by Rel proteins. These are divided into bifunctional and monofunctional proteins based on the presence or absence of the hydrolysis activity. Although these proteins require Mg(2+) for (p)ppGpp synthesis, high Mg(2+) was shown to inhibit this reaction in bifunctional Rel proteins from Mycobacterium tuberculosis and Streptococcus equisimilis. This is not a characteristic feature in enzymes that use a dual metal ion mechanism, such as DNA polymerases that are known to carry out a similar pyrophosphate transfer reaction. Comparison of polymerase Polbeta and Rel(Seq) structures that share a common fold led to the proposal that the latter would follow a single metal ion mechanism. Surprisingly, in contrast to bifunctional Rel, we did not find inhibition of guanosine 5'-triphosphate, 3'-diphosphate (pppGpp) synthesis at higher Mg(2+) in the monofunctional RelA from Escherichia coli. We show that a charge reversal in a conserved motif in the synthesis domains explains this contrast; an RXKD motif in the bifunctional proteins is reversed to an EXDD motif. The differential response of these proteins to Mg(2+) could also be noticed in fluorescent nucleotide binding and circular dichroism experiments. In mutants where the motifs were reversed, the differential effect could also be reversed. We infer that although a catalytic Mg(2+) is common to both bifunctional and monofunctional proteins, the latter would utilize an additional metal binding site formed by EXDD. This work, for the first time, brings out differences in (p)ppGpp synthesis by the two classes of Rel proteins. PMID- 17911109 TI - Role of the diacylglycerol kinase alpha-conserved domains in membrane targeting in intact T cells. AB - Diacylglycerol kinase (DGK) phosphorylates diacylglycerol to phosphatidic acid, modifying the cellular levels of these two lipid mediators. Ten DGK isoforms, grouped into five subtypes, are found in higher organisms. All contain a conserved C-terminal domain and at least two cysteine-rich motifs of unknown function. DGKalpha is a type I enzyme that acts as a negative modulator of diacylglycerol-based signals during T cell activation. Here we studied the functional role of the DGKalpha domains using mutational analysis to investigate membrane binding in intact cells. We show that the two atypical C1 domains are essential for plasma membrane targeting of the protein in intact cells but unnecessary for catalytic activity. We also identify the C-terminal sequence of the protein as essential for membrane binding in a phosphatidic acid-dependent manner. Finally we demonstrate that, in the absence of the calcium binding domain, receptor-dependent translocation of the truncated protein is regulated by phosphorylation of Tyr(335). This functional study provides new insight into the role of the so-called conserved domains of this lipid kinase family and demonstrates the existence of additional domains that confer specific plasma membrane localization to this particular isoform. PMID- 17911110 TI - Internalization of beta-amyloid peptide by primary neurons in the absence of apolipoprotein E. AB - Extracellular accumulation of beta-amyloid peptide (Abeta) has been linked to the development of Alzheimer disease. The importance of intraneuronal Abeta has been recognized more recently. Although considerable evidence indicates that extracellular Abeta contributes to the intracellular pool of Abeta, the mechanisms involved in Abeta uptake by neurons are poorly understood. We examined the molecular mechanisms involved in Abeta-(1-42) internalization by primary neurons in the absence of apolipoprotein E. We demonstrated that Abeta-(1-42) is more efficiently internalized by axons than by cell bodies of sympathetic neurons, suggesting that Abeta-(1-42) uptake might be mediated by proteins enriched in the axons. Although the acetylcholine receptor alpha7nAChR, previously suggested to be involved in Abeta internalization, is enriched in axons, our results indicate that it does not mediate Abeta-(1-42) internalization. Moreover, receptors of the low density lipoprotein receptor family are not essential for Abeta-(1-42) uptake in the absence of apolipoprotein E because receptor-associated protein had no effect on Abeta uptake. By expressing the inactive dynamin mutant dynK44A and the clathrin hub we found that Abeta-(1-42) internalization is independent of clathrin but dependent on dynamin, which suggests an endocytic pathway involving caveolae/lipid rafts. Confocal microscopy studies showing that Abeta did not co-localize with the early endosome marker EEA1 further support a clathrin-independent mechanism. The lack of co localization of Abeta with caveolin in intracellular vesicles and the normal uptake of Abeta by neurons that do not express caveolin indicate that Abeta does not require caveolin either. Instead partial co-localization of Abeta-(1-42) with cholera toxin subunit B and sensitivity to reduction of cellular cholesterol and sphingolipid levels suggest a caveolae-independent, raft-mediated mechanism. Understanding the molecular events involved in neuronal Abeta internalization might identify potential therapeutic targets for Alzheimer disease. PMID- 17911111 TI - Correctors promote maturation of cystic fibrosis transmembrane conductance regulator (CFTR)-processing mutants by binding to the protein. AB - The most common cause of cystic fibrosis (CF) is defective folding of a cystic fibrosis transmembrane conductance regulator (CFTR) mutant lacking Phe(508) (DeltaF508). The DeltaF508 protein appears to be trapped in a prefolded state with incomplete packing of the transmembrane (TM) segments, a defect that can be repaired by expression in the presence of correctors such as corr-4a, VRT-325, and VRT-532. To determine whether the mechanism of correctors involves direct interactions with CFTR, our approach was to test whether correctors blocked disulfide cross-linking between cysteines introduced into the two halves of a Cys less CFTR. Although replacement of the 18 endogenous cysteines of CFTR with Ser or Ala yields a Cys-less mutant that does not mature at 37 degrees C, we found that maturation could be restored if Val(510) was changed to Ala, Cys, Ser, Thr, Gly, Ala, or Asp. The V510D mutation also promoted maturation of DeltaF508 CFTR. The Cys-less/V510A mutant was used for subsequent cross-linking analysis as it yielded relatively high levels of mature protein that was functional in iodide efflux assays. We tested for cross-linking between cysteines introduced into TM6 and TM7 of Cys-less CFTR/V510A because cross-linking between TM6 and TM7 of P glycoprotein, the sister protein of CFTR, was inhibited with the corrector VRT 325. Cys-less CFTR/V510A mutant containing cysteines at I340C(TM6) and S877C(TM7) could be cross-linked with a homobifunctional cross-linker. Correctors and the CFTR channel blocker benzbromarone, but not P-glycoprotein substrates, inhibited cross-linking of mutant I340C(TM6)/S877C(TM7). These results suggest that corrector molecules such as corr-4a interact directly with CFTR. PMID- 17911112 TI - Evidence for the direct involvement of the proteasome in the proteolytic processing of the Aspergillus nidulans zinc finger transcription factor PacC. AB - The 72-kDa zinc finger transcription factor PacC, distantly related to Ci/Gli developmental regulators, undergoes two-step proteolytic processing in response to alkaline ambient pH. "Signaling protease" cleavage of PacC(72) removes a processing-inhibitory C-terminal domain, making its truncated PacC(53) product accessible to a second "processing" protease, yielding PacC(27). Features of the processing proteolysis suggested the proteasome as a candidate protease. We constructed, using gene replacements, two missense active site mutations in preB, the Aspergillus nidulans orthologue of Saccharomyces cerevisiae PRE2 encoding the proteasome beta5 subunit. preB1(K101A) is lethal. Viable preB2(K101R) impairs growth and, like its equivalent pre2(K108R) in yeast, impairs chymotryptic activity. pre2(K108R) and preB2(K101R) active site mutations consistently shift position of the scissile bonds when PacC is processed in S. cerevisiae and A. nidulans, respectively, indicating that PacC must be a direct substrate of the proteasome. preB2(K101R) leads to a 2-3-fold elevation in NimE mitotic cyclin levels but appears to result in PacC instability, suggesting an altered balance between processing and degradation. preB2(K101R) compensates the marked impairment in PacC(27) formation resulting from deletion of the processing efficiency determinant in PacC, further indicating direct proteasomal involvement in the formation of PacC(27). Deletion of a Gly-Pro-Ala-rich region within this processing efficiency determinant markedly destabilizes PacC. Arg substitutions of Lys residues within this efficiency determinant and nearby show that they cooperate to promote PacC processing. A quadruple Lys-to-Arg substitution (4K- >R) impairs formation of PacC(27) and leads to persistence of PacC(53). Wild-type PacC(53) becomes multiply phosphorylated upon alkaline pH exposure. Processing impaired 4K-->R PacC(53) becomes excessively phosphorylated. PMID- 17911113 TI - hCTR9, a component of Paf1 complex, participates in the transcription of interleukin 6-responsive genes through regulation of STAT3-DNA interactions. AB - PAF, which is composed of Paf1, Cdc73, Ctr9, Leo1, and Rtf1, is a novel complex with multiple functions in transcription-related activities. The PAF complex interacts with histone-modifying enzymes and RNA polymerase II to regulate transcription. With general transcription regulatory potential in yeast, Hyrax/Cdc73 has been reported to associate with beta-catenin to control Wnt/Wg signal-specific transcription in Drosophila. Here, we present the first evidence of IL-6 signal-specific transcriptional regulation by SH2BP1/CTR9 in mammals. Upon LPS injection of mice, we observed transient induction of the mammalian PAF complex in the liver. Inhibition of CTR9 specifically abrogated expression of IL 6-responsive genes, but had no effect on genes constitutively expressed or induced by interferon-beta, TNFalpha, or IL-1beta. The PAF complex was found in the promoter regions of IL-6-responsive HP and FGGgamma, but not in the promoter region of constitutively active GAPDH. Transcriptional activation by STAT3 was inhibited when CTR9 siRNA was introduced, whereas transcriptional activation was enhanced by mCtr9 overexpression. IL-6-activated Stat3 was found to co-localize and interact with CTR9. In CTR9-depleted cells, decreased STAT3 association with the promoter regions, as well as impaired K4-trimethylation of histone H3 in the coding regions, of target genes was observed. These data suggest that CTR9 participates in the transcription of IL-6-responsive genes through the regulation of DNA association of STAT3 and modification of histone methylation. PMID- 17911114 TI - Injection of flagellin into the host cell cytosol by Salmonella enterica serotype Typhimurium. AB - Bacterial flagellins are potent inducers of innate immunity. Three signaling pathways have been implicated in the sensing of flagellins; these involve toll like receptor 5 (TLR5) and the cytosolic proteins Birc1e/Naip5 and Ipaf. Although the structural basis of TLR5-flagellin interaction is known, little is known about how flagellin enters the host cell cytosol to induce signaling via Birc1e/Naip5 and Ipaf. Here we demonstrate for the first time the translocation of bacterial flagellin into the cytosol of host macrophages by the vacuolar pathogen, Salmonella enterica serotype Typhimurium. Translocation of flagellin into the host cell cytosol was directly demonstrated using beta-lactamase reporter constructs. Flagellin translocation required the Salmonella Pathogenicity Island 1 Type III secretion system (SPI-1 T3SS) but not the flagellar T3SS. PMID- 17911115 TI - A NICE debate likely to turn nasty. PMID- 17911116 TI - Addressing uncertainties about the effects of treatments offered to NHS patients: whose responsibility? PMID- 17911117 TI - A NICE mess. PMID- 17911118 TI - Adverse effects of spinal manipulation. PMID- 17911119 TI - Adverse effects of spinal manipulation. PMID- 17911120 TI - Adverse effects of spinal manipulation. PMID- 17911121 TI - Adverse effects of spinal manipulation. PMID- 17911122 TI - Adverse effects of spinal manipulation. PMID- 17911123 TI - Adverse effects of spinal manipulation. PMID- 17911124 TI - Adverse effects of spinal manipulation. PMID- 17911126 TI - Implementing NICE guidance. PMID- 17911127 TI - Making a decision to wait for more evidence: when the National Institute for Health and Clinical Excellence recommends a technology only in the context of research. PMID- 17911128 TI - Policy on the prevention of suicidal behaviour; one treatment for all may be an unrealistic expectation. AB - In the absence of clear guidelines on suicide prevention there is a pressing need to translate existing and future evidence into policy. Suicide is a behaviour, not a diagnosis, and has diverse underlying pathologies. Interventions have differential effects in specific risk groups, which may include paradoxical increases in risk. For these reasons, policy makers may need to abandon the goal of one treatment for all and focus on the distinct subgroups of patients at risk when selecting, evaluating and implementing preventive interventions. This has implications for the design of future research, but has the potential to increase the utility and cost-effectiveness of the data available, thereby benefiting policy makers, clinicians and patients. PMID- 17911129 TI - Contraception and abortion. PMID- 17911130 TI - Insights into how to conduct a clinical trial in the UK. AB - New researchers may find starting and conducting clinical studies in the UK complicated and time-consuming. In this article, we describe our collective experiences and provide some pointers on how to navigate through the various committees and regulatory bodies. The article is intended to aid junior researchers in understanding the study process and to provide them with some insight on how to get through this complex system successfully. PMID- 17911131 TI - Age-related macular degeneration: what do patients find on the internet? AB - OBJECTIVE: To assess the quality of information and readability of the top internet sites for age-related macular degeneration (AMD). DESIGN: An examination of the technical information provision, quality and readability of websites found during an internet search for 'age-related macular degeneration'. Setting Six internet search engines were used to find 26 unique sites on AMD. MAIN OUTCOME MEASURES: Technical information and quality were assessed using a simple grading system. Readability was assessed using a Simple Measure Of Gobbledygook (SMOG) rating. RESULTS: Twelve organizational, seven academic and seven commercial sites were identified. The average technical scores were 82.3%, 67.9% and 65.2% for each type of site, respectively (P=0.097, one way ANOVA). The average quality scores were 62.2%, 62.6%, and 49.5% for each type of site, respectively (P=0.356, one-way ANOVA). The average SMOG ratings were 16.3, 16.1, and 16.2 for each type of site, respectively (P=0.983, one-way ANOVA). Fifteen of the sites provided details of new and emerging treatments, with seven providing a detailed discussion. CONCLUSIONS: Many websites are now meeting the challenge of providing comprehensive information about AMD and its new treatments. Quality scores were disappointing, with sites needing to provide more evidence of authorship and attribution of information. The majority of sites had SMOG scores above 10, making them difficult for the average person to understand. As physicians we need to help design and direct patients to sites that provide high quality, current information. PMID- 17911132 TI - A funny patient. PMID- 17911133 TI - An 18th century controlled trial prompted by a potential shortage of hospital beds. PMID- 17911134 TI - The media and cancer survival. PMID- 17911135 TI - Chlamydia trachomatis seroprevalence atlas of Finland 1983-2003. AB - OBJECTIVES: To study Chlamydia trachomatis seroprevalence trends and geographical distribution over time in Finland. MATERIALS AND METHODS: First pregnancy serum samples were retrieved from the Finnish Maternity Cohort serum bank for the subcohort of 8000 women stratified by calendar years (1983-1989, 1990-1996, 1997 2003) and age at time of sample withdrawal (14-22 and 23-28 years). C trachomatis antibodies were determined using standard major outer membrane protein peptide ELISA. The spatiotemporal variation of C trachomatis seroprevalence rates was visualised by a series of maps. RESULTS: A decreasing C trachomatis seroprevalence trend from 1983 to 2003 was seen for both women under 23 years of age (20.8% to 10.6%) and 23-28-year-old women (19.1% to 12.5%). Constant clusters were seen around the largest cities and in eastern Finland although seroprevalence rates were generally decreasing throughout the country. CONCLUSIONS: Only a few population-based serological studies have been undertaken on C trachomatis epidemiology over time. In Finland the seroprevalence of C trachomatis is decreasing all over the country, albeit with small clusters remaining. PMID- 17911136 TI - HIV voluntary counselling and testing for African communities in London: learning from experiences in Kenya. AB - OBJECTIVES: To explore the feasibility and acceptability of translating a successful voluntary counselling and testing (VCT) service model from Kenya to African communities in London. METHODS: Qualitative study with focus group discussions and a structured workshop with key informants. Five focus group discussions were conducted in London with 42 participants from 14 African countries between August 2006 and January 2007. A workshop was held with 28 key informants. Transcripts from the group discussions and workshop were analysed for recurrent themes. RESULTS: Participants indicated that a community-based HIV VCT service would be acceptable to African communities in London, but also identified barriers to uptake: HIV-related stigma, concerns about confidentiality, and doubts about the ability of community-based services to maintain professional standards of care. Workshop participants highlighted three key requirements to ensure feasibility: (a) efficient referrals to sexual health services for the newly diagnosed; (b) a locally appropriate testing algorithm and quality assurance scheme; (c) a training programme for VCT counsellors. CONCLUSIONS: Offering community-based VCT with rapid HIV tests appears feasible within a UK context and acceptable to African communities in London, provided that clients' confidentiality is ensured and appropriate support is given to the newly diagnosed. However, the persistence of concerns related to HIV-related stigma among African communities suggests that routine opt-out testing in healthcare settings may also constitute an effective approach to reducing the proportion of late diagnoses in this group. HIV service models and programmes from Africa constitute a valuable knowledge base for innovative interventions in other settings, including developed countries. PMID- 17911137 TI - Concordance of chlamydia trachomatis infections within sexual partnerships. AB - OBJECTIVES: The enhanced sensitivity of nucleic acid amplification tests (NAAT) provides an opportunity for estimating the prevalence of untreated Chlamydia trachomatis infections. The transmissibility and public health significance of some NAAT-identified infections are, however, not known. METHODS: Adults attending an urban emergency department provided specimens for C trachomatis screening using NAAT. Participants testing positive were offered follow-up including re-testing for C trachomatis using NAAT and traditional methods, eg culture and direct fluorescent antibody, and were treated. Partners were offered identical evaluation and treatment. Overall, 90 C trachomatis-positive participants had one or more sexual partners enrolled. RESULTS: Evidence of transmission, as defined by infection concordance between partnerships, was observed among 75% of partners of index cases testing positive by both NAAT and traditional assay but only 45% of partners of index cases testing positive by NAAT only (prevalence ratio 1.7, 95% CI 1.1 to 2.5). Among index participants returning for follow-up, 17% had no evidence of C trachomatis infection by NAAT or traditional assay (median follow-up three weeks). CONCLUSIONS: A substantial proportion of positive NAAT results for chlamydial infection may be of lower transmissibility and may not persist after a short follow-up. The long-term health effects of some positive NAAT are uncertain. PMID- 17911138 TI - Hormonal contraceptive use modulates the local inflammatory response to bacterial vaginosis. AB - OBJECTIVES: To compare cervical concentrations of numerous cytokines/chemokines in women with bacterial vaginosis (BV) compared with the levels detected after BV resolution and determine if hormonal contraceptive use modulates the local inflammatory response to BV. METHODS: Cervical secretions from 81 women with BV at enrollment and normal flora at one-month follow-up were analysed for 10 different cytokines/chemokines using multiplexed fluorescent bead-based immunoassays. RESULTS: BV was associated with significantly higher concentrations of IL-1 beta, tumour necrosis factor (TNF), interferon-gamma, IL-2, IL-4, and IL 10 compared with the levels detected in the presence of normal vaginal flora. Analysis of results stratified by contraceptive practice demonstrated significantly lower levels of numerous cytokines among women with BV using hormonal contraceptives compared with those women with BV not using hormonal contraceptives. Hormonal contraceptive use was also associated with a statistically significant lesser change in TNF levels between the two study visits compared with the amount of change detected between visits among women who denied their use. CONCLUSIONS: Despite increases in the levels of both pro and anti-inflammatory cytokines in the lower genital tract of women with BV, the overall balance of these two types of molecules was maintained. The character of this local inflammatory response may help explain the typical absence of overt signs of inflammation among women with BV. In addition, hormonal contraceptive use was associated with significantly lower levels of the pro-inflammatory molecules TNF, interferon-gamma, and granulocyte macrophage colony-stimulating factor in women with BV, but did not significantly reduce the levels of IL-10, a key anti-inflammatory cytokine. These results suggest the possibility of an association between hormonal contraceptive use and altered genital tract immunity. PMID- 17911139 TI - Cost effectiveness of one to one STI prevention interventions. PMID- 17911140 TI - Elusive aetiology of bacterial vaginosis. Do lesbians have a clue? PMID- 17911141 TI - Direct transmission: introducing a new feature in STI. PMID- 17911142 TI - Schedules for hepatitis B vaccination of risk groups: balancing immunogenicity and compliance. AB - INTRODUCTION: Vaccination is an important tool in hepatitis B prevention. However, several vaccine doses are required to induce long-term protection. Several at-risk groups have difficulties in adhering to the standard vaccination schedule. OBJECTIVES: This paper aims to review the use of accelerated hepatitis B vaccination schedules, in terms of immunogenicity and compliance. RESULTS: Accelerated schedules (0.1.2.12 months) or super-accelerated schedules (0.7.21.360 days) have been shown to result in higher proportions of healthy vaccinees reaching anti-HBs antibody levels >or=10 IU/l more rapidly. A fourth completing dose is required to lift antibody levels to an equal height, as does a standard (0.1.6 months) schedule. Accelerated schedules do also increase the uptake of hepatitis B vaccine, that is the proportion of vaccinees who receive three doses. However, completing the schedule with a fourth dose is usually more difficult than completing a standard 0.1.6-month schedule. Several additional tools can help to increase the compliance (eg, reminder systems, outreach services and incentive schemes). CONCLUSION: For rapid seroconversion and almost immediate protection in the short term, a (super)accelerated schedule could be used in at-risk groups. As long-term protection data with these (super) accelerated schedules have not been documented yet, a fourth dose at month 12 is still required. A shortened schedule (0.1.4 months) might be an alternative worth considering compared with the standard 0.1.6, as it convenes to internationally accepted minimum dose intervals and offers earlier protection. There is a clear need to study the long-term protection and effectiveness of the primary part of (super)accelerated schedules. PMID- 17911143 TI - Data from UK genitourinary medicine clinics, 2006: a mixed picture. AB - OBJECTIVE: To give an overview of the latest latest trends in diagnoses made and services provided by genitourinary medicine (GUM) clinics in the UK. METHODS: Aggregate data collected from the KC60 statistical returns for GUM clinics in England, Wales and Northern Ireland and disaggregate data collected using the STI Surveillance System for GUM Clinics in Scotland. These data were collated and numbers of diagnoses were adjusted for missing clinic data. RESULTS & CONCLUSION: Overall, numbers of new diagnoses of sexually transmitted infections (STIs) continued to rise in 2006. However, there was some evidence of improvement, with new diagnoses of gonorrhoea falling for the fourth successive year. Chlamydia continued to be the most common STI diagnosed in GUM clinics, and the sharp rise in new diagnoses over the last 10 years was most likely associated with an increase in testing volume and accuracy. The highest rates of STI diagnoses continued, in the main, to be among 16-24-year-olds, and there were some notable rises among this age group also: new diagnoses of genital herpes in teenage women rose by 16% in 2006. Improving the sexual health of men who have sex with men (MSM) must remain a priority, as the increase in numbers of new STI diagnoses among MSM over the past 10 years continued unabated into 2006. However, despite facing the challenge of reducing patient waiting times, there has been a considerable rise in sexual health screens and HIV tests being provided by GUM services, and this could, if sustained, result in significant improvements in sexual health in the coming years. PMID- 17911144 TI - Interview with Peter Piot. Interview by Marc Vandenbruaene. PMID- 17911145 TI - The extent of self-initiated STI and HIV prophylaxis (auto-PEP) and treatment in MSM attending GUM and HIV clinic services. PMID- 17911146 TI - Simvastatin co-prescribed with protease inhibitors despite dangerous drug interactions. PMID- 17911147 TI - Gender transformation requires population approaches to addressing gender-based violence and HIV. PMID- 17911148 TI - Differences in disease prevalence as a source of the U.S.-European health care spending gap. AB - The United States spends more on health care than any European country. Previous studies have sought to explain these differences in terms of system capacity, access to technologies, gross domestic product, and prices. We examine differences in disease prevalence and treatment rates for ten of the most costly conditions between the United States and ten European countries using surveys of the noninstitutionalized population age fifty and older. Disease prevalence and rates of medication treatment are much higher in the United States than in these European countries. Efforts to reduce the U.S. prevalence of chronic illness should remain a key policy goal. PMID- 17911149 TI - Accuracy of child morbidity data in demographic and health surveys. AB - BACKGROUND: The Demographic and Health Surveys (DHSs) have been used throughout the developing world for the last 20 years to provide data on the distribution of disease in order to inform planning. Data on child illness and death are reported by mothers and are susceptible to error. METHODS: We conducted an in-depth study of the Iranian DHS carried out in 2000-2001 and reviewed 110 DHS carried out around the world to check for bias by assessing the social gradient in reported child morbidity and mortality. RESULTS: We found that the reported under-5 child morbidity and mortality rates for the 28 Iranian provinces were inversely correlated (r = -0.592, P < 0.001) and that the adjusted social gradient of child morbidity implied increased illness in those who had literate vs illiterate mothers (OR = 1.26, 95% CI 1.20-1.32) compared with a decrease in mortality with increased literacy (OR = 0.52, 95% CI 0.46-0.59). Many of the other DHSs also show increased rates of reported child diarrhoea in households with higher levels of maternal education, access to piped water and urban (vs rural) dwellings, the reverse of what is found with mortality rates. CONCLUSIONS: This suggests that there may be significant recall and reporting bias in under-5 childhood morbidity in DHSs. Caution should be used in the interpretation and use of data from DHSs and the survey methods should be reviewed. PMID- 17911150 TI - Commentary: strengthening the reporting of observational epidemiology the STROBE statement. PMID- 17911151 TI - Commentary: preventing suicide: need for a life course approach. PMID- 17911152 TI - The impact of airborne dust on respiratory health in children living in the Aral Sea region. AB - BACKGROUND: Anecdotally, people living in the Aral Sea region report an increase in the prevalence of respiratory illnesses, particularly in children, and there is widespread belief that this is due to dust from the Aral Sea bed. METHODS: We conducted a survey of respiratory symptoms and lung function in children aged 7 10 years living in 18 communities in 6 geographical regions in the Aral Sea area. We monitored dust deposition rates monthly for the duration of the study. RESULTS: The overall prevalence of recent wheeze was low at 4.2%, but this figure varied with region and was higher in the more accessible urban and delta regions and lower in the more remote regions. We found no evidence of an association between local annual dust deposition and specific respiratory symptoms. Lung function results also showed variation between geographical regions not explained by annual dust deposition. After allowing for region of residence, however, there was some evidence of an inverse association between percentage predicted forced expiratory volume in 1 s(FEV1) and dust exposure during the summer months (change in percentage predicted FEV1 per 1,000 kg/ha annual dust deposition -1.465) (95% confidence interval -2.519 to -0.412); however, in winter, the reverse was true. CONCLUSIONS: The prevalence of asthma is low in the Aral Sea area and appears to be unrelated to dust exposure. Exposure to dust did not explain the main variations in lung function between geographical regions but high levels of dust exposure during the summer may have an adverse effect on lung function. PMID- 17911153 TI - Cohort profile: The Thai Cohort of 87,134 Open University students. PMID- 17911154 TI - Cohort profile: the Leicester respiratory cohorts. PMID- 17911155 TI - Polyglutamine diseases: emerging concepts in pathogenesis and therapy. AB - Polyglutamine diseases are a family of neurodegenerative conditions that each derive from a CAG triplet repeat expansion in a specific gene. This produces a pathogenic protein that contains a critically expanded tract of glutamines. These prototypical protein misfolding disorders include Huntington disease, spinobulbar muscular atrophy, dentatorubral-pallidoluysian atrophy and several spinocerebellar ataxias. This article reviews the emerging concepts in pathogenesis and therapy. Key ideas include the role of proteolytic cleavage, the importance of conformational change in the pathogenic proteins, the role of protein aggregation and the importance of transcriptional and metabolic disturbances. The relative role of functional perturbation in a target protein induced by a polyglutamine expansion is also discussed. Therapeutic strategies include counteracting cellular perturbations and direct targeting of polyglutamine protein expression, cleavage or conformation. PMID- 17911156 TI - Fruit flies and the sperm proteome. AB - Sperm have been studied for their obvious role in fertilization and as a model system for cell-cell interactions and cell signaling. Despite its central and critical role in reproduction, we know surprisingly little about the overall molecular composition of sperm. Interest in sperm function has greatly intensified for two reasons: first, it is becoming increasingly apparent that human infertility can be traced to male factors, including alterations in sperm proteins, and second, there is increasing empirical evidence that sperm provide essential factors, both nucleic acid- and protein-based, to early zygote development possibly beyond their role in fertilization. At the molecular level, study of the sperm proteome has revealed a variety of genetic mechanisms involved in the organization and evolution of sperm form and function. These discoveries are being augmented and expanded by the application of proteomics that directly identifies protein constituents of sperm. In this article I argue that sperm are ideal candidate cell types for proteomic analyses and describe the current state of the field focussing on the recently described sperm proteome in the fruit fly Drosophila melanogaster. PMID- 17911157 TI - Challenges in human genetic diversity: demographic history and adaptation. AB - Modern human genetic diversity is the result of demographic history, and selective effects that have acted to adapt different populations to their environments. Broad patterns of global diversity are well explained by geography, based on an out-of-Africa model of early human evolution. Genome-wide searches for signals of selection, plus studies of specific candidate loci and candidate phenotypes, have identified genes that show population differences due to adaptation to pathogens, climate, diet and possibly cognitive challenges. Some past adaptations are now maladaptive, and can lead to disease. However, the history of adaptation is complex, and adaptive explanations are often unsupported by hard evidence. PMID- 17911158 TI - The necessary junk: new functions for transposable elements. AB - Transposable elements have been shaping the genome throughout evolution, contributing to the creation of new genes and sophisticated regulatory network systems. Today, most of genomes (animals and plants) allow the expression and accommodate transposition of a few transposon families. The potential genetic impact of this small fraction of mobile elements should not be underestimated. Although new insertions that happen in germ cells are likely to be passed to the next generation, mobilization in pluripotent embryonic stem cells or in somatic cells may contribute to the differences observed in genetic makeup and epigenetic gene regulation during development at the cellular level. The fact that these elements are still active, generating innovative ways to alter gene expression and genomic structure, suggests that the cellular genome is not static or deterministic but rather dynamic. In this short review, we collect a set of recent observations that point to a new appreciation of transposable elements as a source of genetic variation. PMID- 17911159 TI - Copy-number variation in control population cohorts. AB - Copy-number variation (CNV) is the most prevalent type of structural variation in the human genome, and contributes significantly to genetic heterogeneity. It has already been recognized that some CNVs can contribute to human phenotype, including rare genomic disorders and Mendelian diseases. Other CNVs are now amenable to genome-wide association studies so that their influence on human phenotypic diversity and disease susceptibility may soon be more readily determined. Population studies and reference databases for control and disease associated samples are required to provide an information resource about CNV frequencies and their relative contribution to phenotypic outcomes. The relatively high cost of screening individual samples has tended to limit the number of controls assayed, and use of the data has often been hampered by the variety of technology platforms and analysis techniques. As a result, there is still a paucity of data on population frequency and distribution of CNVs, particularly for those that are rare. Here, we provide an example of how to discover new CNVs from existing genotype data from large-scale genetic epidemiological studies. We also discuss the need to expand surveys of CNV in different population-based cohorts and to apply the information to studies of human variation and disease. PMID- 17911160 TI - Genetic susceptibility to age-related macular degeneration: a paradigm for dissecting complex disease traits. AB - Age-related macular degeneration (AMD) is a progressive neurodegenerative disease, which affects quality of life for millions of elderly individuals worldwide. AMD is associated with a diverse spectrum of clinical phenotypes, all of which include the death of photoreceptors in the central part of the human retina (called the macula). Tremendous progress has been made in identifying genetic susceptibility variants for AMD. Variants at chromosome 1q32 (in the region of CFH) and 10q26 (LOC387715/ARMS2) account for a large part of the genetic risk to AMD and have been validated in numerous studies. In addition, susceptibility variants at other loci, several as yet unidentified, make substantial cumulative contribution to genetic risk for AMD; among these, multiple studies support the role of variants in APOE and C2/BF genes. Genome wide association and re-sequencing projects, together with gene-environment interaction studies, are expected to further define the causal relationships that connect genetic variants to AMD pathogenesis and should assist in better design of prevention and intervention. PMID- 17911161 TI - Parkinson's disease. AB - Parkinson's disease (PD) is a chronic progressive neurodegenerative movement disorder characterized by a profound and selective loss of nigrostriatal dopaminergic neurons. Clinical manifestations of this complex disease include motor impairments involving resting tremor, bradykinesia, postural instability, gait difficulty and rigidity. Current medications only provide symptomatic relief and fail to halt the death of dopaminergic neurons. A major hurdle in development of neuroprotective therapies are due to limited understanding of disease processes leading to death of dopaminergic neurons. While the etiology of dopaminergic neuronal demise is elusive, a combination of genetic susceptibilities and environmental factors seems to play a critical role. The majority of PD cases are sporadic however, the discovery of genes linked to rare familial forms of disease (encoding alpha-synuclein, parkin, DJ-1, PINK-1 and LRRK2) and studies from experimental animal models has provided crucial insights into molecular mechanisms in disease pathogenesis and identified probable targets for therapeutic intervention. Recent findings implicate mitochondrial dysfunction, oxidative damage, abnormal protein accumulation and protein phosphorylation as key molecular mechanisms compromising dopamine neuronal function and survival as the underlying cause of pathogenesis in both sporadic and familial PD. In this review we provide an overview of the most relevant findings made by the PD research community in the last year and discuss how these significant findings improved our understanding of events leading to nigrostriatal dopaminergic degeneration, and identification of potential cell survival pathways that could serve as targets for neuroprotective therapies in preventing this disabling neurological illness. PMID- 17911162 TI - Genetic basis of polygenic hypertension. AB - Essential hypertension is a common disorder that leads to significant morbidity and mortality; however, the underlying mechanisms have remained elusive. Recent animal model studies have uncovered a complex genetic architecture of quantitative trait loci (QTLs) for blood pressure (BP), intricate QTL-QTL interactions and powerful genome regulations that underlie polygenic hypertension. BP, a quantitative trait manifesting as a continuous variation, seems to be controlled by individual 'monogenic' QTLs following Mendelian inheritance. Certain QTLs are functionally organized in epistatic modules that likely participate in pathways and cascades, whereas others belong to independent modules. This understanding provides insights into probable genetic mechanisms underlying essential hypertension. Translation of gene discovery to therapy will require an integrated approach that includes experimental validation of genes in animal models and in humans. PMID- 17911163 TI - The origin of human aneuploidy: where we have been, where we are going. AB - Aneuploidy is the most common chromosome abnormality in humans, and is the leading genetic cause of miscarriage and congenital birth defects. Since the identification of the first human aneuploid conditions nearly a half-century ago, a great deal of information has accrued on its origin and etiology. We know that most aneuploidy derives from errors in maternal meiosis I, that maternal age is a risk factor for most, if not all, human trisomies, and that alterations in recombination are an important contributor to meiotic non-disjunction. In this review, we summarize some of the data that have led to these conclusions, and discuss some of the approaches now being used to address the underlying causes of meiotic non-disjunction in humans. PMID- 17911164 TI - Insights from spatially mapped gene expression in the mouse brain. AB - The growing number of publicly available databases of murine gene expression arising from genomic-scale transcriptome/proteome profiling projects allows open access to information about genes potentially involved in diseases and disorders of the brain. The use of various methodologies by myriad projects provides complementary types of information, ranging from easily quantifiable microarray data for gross brain regions, to transcript tag analysis and proteomic characterization. One mode of gene expression analysis that has recently been widely adopted is the utilization of colorimetric in situ hybridization. This approach is adaptable for high throughput production, and provides a reproducible, scaleable platform for large datasets. The Allen Brain Atlas in particular has utilized this technology to produce a genomic-scale anatomical digital atlas of gene expression in the adult male mouse brain. The availability of global datasets with cellular level spatial resolution, which can be easily parsed due to accessible informatics-derived image analysis tools, can provide both high level and detailed insights into gene regulation. This article reviews various gene expression profiling projects in the mouse brain, how these data sets are increasingly used to complement other studies and applications of these datasets to further understanding of neurological disease. PMID- 17911165 TI - Genomic strategies for personalized cancer therapy. AB - Numerous recent studies have demonstrated the use of genomic data, particularly gene expression signatures, as clinical prognostic factors in cancer and other complex diseases. These studies highlight the opportunity for strategies to achieve truly personalized cancer treatment. In particular, the ability to develop gene expression signatures will likely allow us to guide the use of currently available cancer drugs, develop new targeted therapeutics, and provide an opportunity to better match the most effective drug or drugs with the molecular characteristics of the individual patient. PMID- 17911166 TI - Genetics of sporadic amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized clinically by rapidly progressive paralysis leading ultimately to death from respiratory failure. There is substantial evidence suggesting that ALS is a heritable disease, and a number of genes have been identified as being causative in familial ALS. In contrast, the genetics of the much commoner sporadic form of the disease is poorly understood and no single gene has been definitively shown to increase the risk of developing ALS. In this review, we discuss the genetic evidence for each candidate gene that has been putatively associated with increased risk of sporadic ALS. We also review whole genome association studies of ALS and discuss the potential of this methodology for identifying genes relevant to motor neuron degeneration. PMID- 17911167 TI - Status of genomic imprinting in human embryonic stem cells as revealed by a large cohort of independently derived and maintained lines. AB - Investigation of the epigenetic stability of human embryonic stem cells (hESCs) is a crucial step for their use in cell-replacement therapies, as well as for assessing whether hESCs model epigenetic regulation in human pre-implantation cell types. To address these issues, we have examined the expression of imprinted genes in a previous study and more recently in 46 individual hESC lines as part of the International Stem Cell Initiative. Our results show that nearly all hESC lines examined possessed a substantial degree of epigenetic stability, despite differences in genetic background and in their derivation and initial propagation conditions. However, some hESCs did show loss of allele-specific expression, which could have implications for hESC differentiation and epigenetic stability (both in vitro and after clinical transplantation). A benefit of our and other recent studies of genomic imprinting in hESCs was the identification of imprinted genes that provide a useful indication of epigenetic stability. SNRPN, IPW and KCNQ1OT1 were highly stable and thus appeared insensitive to perturbation; in contrast, H19, IGF2 and MEG3 were more variable and thus could potentially provide a sensitive indication of epigenetic status. In this review, we examine the differences between imprinted genes in their susceptibility to perturbation and discuss the potential molecular basis for these differences. This examination provides insight into the regulation of genomic imprinting in hESCs and the corresponding peri-implantation stages of human development. PMID- 17911169 TI - alpha6beta4 integrin activates Rac-dependent p21-activated kinase 1 to drive NF kappaB-dependent resistance to apoptosis in 3D mammary acini. AB - Malignant transformation and multidrug resistance are linked to resistance to apoptosis, yet the molecular mechanisms that mediate tumor survival remain poorly understood. Because the stroma can influence tumor behavior by regulating the tissue phenotype, we explored the role of extracellular matrix signaling and tissue organization in epithelial survival. We report that elevated (alpha6)beta4 integrin-dependent Rac-Pak1 signaling supports resistance to apoptosis in mammary acini by permitting stress-dependent activation of the p65 subunit of NF-kappaB through Pak1. We found that inhibiting Pak1 through expression of N17Rac or PID compromises NF-kappaB activation and renders mammary acini sensitive to death, but that resistance to apoptosis could be restored to these structures by overexpressing wild-type NF-kappaB p65. We also observed that acini expressing elevated levels of Pak1 can activate p65 and survive death treatments, even in the absence of activated Rac, yet will die if activation of NF-kappaB is simultaneously inhibited through expression of IkappaBalphaM. Thus, mammary tissues can resist apoptotic stimuli by activating NF-kappaB through alpha6beta4 integrin-dependent Rac-Pak1 signaling. Our data emphasize the importance of the extracellular matrix stroma in tissue survival and suggest that alpha6beta4 integrin-dependent Rac stimulation of Pak1 could be an important mechanism mediating apoptosis-resistance in some breast tumors. PMID- 17911168 TI - Arabidopsis POT1A interacts with TERT-V(I8), an N-terminal splicing variant of telomerase. AB - Chromosome integrity is maintained via the actions of ribonucleoprotein complexes that can add telomeric repeats or can protect the chromosome end from being degraded. POT1 (protection of telomeres 1), a class of single-stranded-DNA binding proteins, is a regulator of telomeric length. The Arabidopsis genome contains three POT1 homologues: POT1A, POT1B and POT1C. Using yeast two-hybrid assays to identify components of a potential POT1A complex, we retrieved three interactors: the N-terminus of the telomerase, a protein kinase and a plant specific protein. Further analysis of the interaction of POT1 proteins with telomerase showed that this interaction is specific to POT1A, suggesting a specific role for this paralogue. The interaction is specific to the N-terminal region of the telomerase, which can be encoded by splicing variants. This interaction indicates possible mechanisms for telomerase regulation by alternative splicing and by POT1 proteins. PMID- 17911170 TI - Congenital growth hormone (GH) deficiency and atherosclerosis: effects of GH replacement in GH-naive adults. AB - BACKGROUND: GH deficiency (GHD) in adults is associated with increased abdominal adiposity and systolic blood pressure, total and low-density lipoprotein cholesterol, and C-reactive protein. METHODS: We have studied the effects of 6 month GH replacement therapy in 20 adult members of a large Brazilian kindred with lifelong severe and isolated GHD due to a homozygous mutation in GHRH receptor gene (46 +/- 14.5 yr; 122 +/- 7.7 cm; 36.7 +/- 5.4 kg; 10 men). Subjects were studied at baseline, after 6-month bimonthly depot GH injections (Nutropin Depot; Genentech, Inc., South San Francisco, CA) [post GH (pGH)], and after 6- and 12-month washout. RESULTS: Despite modest trough serum IGF-I increase, GH replacement therapy caused a decrease in skinfolds and in waist-hip ratio, with a rebound increase at 12 months. Total and low-density lipoprotein cholesterol were reduced pGH and returned to baseline at 6 months. High-density lipoprotein cholesterol increased pGH, but at 12 months was lower than baseline. A progressive increase in left ventricular mass index, posterior wall, and septum thickness occurred from pGH to 12 months, and of carotid intima-media thickness, from 6 to 12 months. Individuals were 6, 16, and 52 times more likely to have an atherosclerotic carotid plaque at pGH, 6 and 12 months, respectively, when compared with baseline. CONCLUSION: In patients with lifetime isolated GHD, 6 month treatment with GH has reversible beneficial effects on body composition and metabolic profile, but it causes a progressive increase in intima-media thickness and in the number of atherosclerotic carotid plaques. PMID- 17911171 TI - Age-specific distribution of serum thyrotropin and antithyroid antibodies in the US population: implications for the prevalence of subclinical hypothyroidism. AB - CONTEXT: Measurements from all age groups defined the upper limit of the TSH reference range in National Health and Nutrition Examination Survey III. The TSH median, 97.5 centile and prevalence of subclinical hypothyroidism (SCH), normal serum T(4) and TSH greater than 4.5 mIU/liter, increased progressively with age. Age-adjusted reference ranges would include many people with TSH greater than 4.5 mIU/liter. OBJECTIVE: We determined whether increasing 50 and 97.5 centiles with age resulted from more patients with SCH in populations with normal TSH distribution or whether age-specific population shifts to higher serum TSH might account for these findings. DESIGN/SETTING/PATIENTS: We analyzed TSH, antithyroid antibodies, and TSH frequency distribution curves for specific age deciles in populations without thyroid disease, with or without antithyroid antibodies. RESULTS: Without thyroid disease, 10.6% of 20- to 29-yr-olds had TSH greater than 2.5 mIU/liter, increasing to 40% in the 80+ group, 14.5% of whom had TSH greater than 4.5 mIU/liter. When TSH was greater than 4.5 mIU/liter, the percentage with antibodies was 67.4% (age 40-49 yr) and progressively decreased to 40.5% in the 80+ group. TSH frequency distribution curves of the 80+ group with or without antibodies was displaced to higher TSH, including TSH at peak frequency. The 97.5 centiles for the 20-29 and 80+ groups were 3.56 and 7.49 mIU/liter, respectively. Seventy percent of older patients with TSH greater than 4.5 mIU/liter were within their age-specific reference range. CONCLUSION: TSH distribution progressively shifts toward higher concentrations with age. The prevalence of SCH may be significantly overestimated unless an age-specific range for TSH is used. PMID- 17911172 TI - Low-grade inflammation, obesity, and insulin resistance in adolescents. AB - CONTEXT: Low-grade inflammation is associated with insulin resistance and precedes the onset of type 2 diabetes mellitus in adults, but there are no comparable data in youth. OBJECTIVE: The objective of the study was to characterize the pattern of subclinical immune activation that is associated with indices of obesity and insulin resistance in youth and analyze whether this association is explained by obesity. DESIGN: This was a cross-sectional study. SETTING: Medical check-up of schoolchildren was conducted by the Public Health Office in Dusseldorf (Germany). PARTICIPANTS: Participants included 519 adolescents (mean age 15.5 +/- 0.8 yr). MAIN OUTCOME MEASURES: Measures included body mass index (BMI) and waist circumference (WC) as indices of obesity; fasting glucose, insulin, and homeostasis model assessment of insulin resistance; serum concentrations of TNFalpha, IL-6, IL-8, IL-18, monocyte chemoattractant protein 1, interferon-gamma-inducible protein (IP)-10 and adiponectin as immunological variables. RESULTS: In age-, sex-, and lipid-adjusted analyses, IL-6, IL-18, IP 10, and adiponectin (inversely) were associated with both BMI and WC (all P 10 but 30 but 0.10). However, it should be noted that because of the limited number of fractures, this study does not exclude the possibility of differences in fracture outcome between the subgroups. CONCLUSIONS: In postmenopausal women with osteoporosis and normal intact PTH, the responses to TPTD did not differ significantly in women with baseline 25OHD insufficiency or sufficiency. PMID- 17911179 TI - Involvement of CCR9 at multiple stages of adult T lymphopoiesis. AB - The chemokine CCL25 is constitutively expressed in the thymus, and its receptor CCR9 is expressed on subsets of developing thymocytes. Nevertheless, the function of CCL25/CCR9 in adult thymopoiesis remains unclear. Here, we demonstrate that purified CCR9(-/-) hematopoietic stem cells are deficient in their ability to generate all major thymocyte subsets including double-negative 1 (DN1) cells in competitive transfers. CCR9(-/-) bone marrow contained normal numbers of lineage( ) Sca-1+c-kit+, common lymphoid progenitors, and lymphoid-primed multipotent progenitors (LMPP), and CCR9(-/-) LMPP showed similar T cell potential as their wild-type (WT) counterparts when cultured on OP9-delta-like 1 stromal cells. In contrast, early thymic progenitor and DN2 thymocyte numbers were reduced in the thymus of adult CCR9(-/-) mice. In fetal thymic organ cultures (FTOC), CCR9(-/-) DN1 cells were as efficient as WT DN1 cells in generating double-positive (DP) thymocytes; however, under competitive FTOC, CCR9(-/-) DP cell numbers were reduced significantly. Similarly, following intrathymic injection into sublethally irradiated recipients, CCR9(-/-) DN cells were out-competed by WT DN cells in generating DP thymocytes. Finally, in competitive reaggregation thymic organ cultures, CCR9(-/-) preselection DP thymocytes were disadvantaged significantly in their ability to generate CD4 single-positive (SP) thymocytes, a finding that correlated with a reduced ability to form TCR-MHC-dependent conjugates with thymic epithelial cells. Together, these results highlight a role for CCR9 at several stages of adult thymopoiesis: in hematopoietic progenitor seeding of the thymus, in the DN-DP thymocyte transition, and in the generation of CD4 SP thymocytes. PMID- 17911180 TI - Expression of human NDRG2 by myeloid dendritic cells inhibits down-regulation of activated leukocyte cell adhesion molecule (ALCAM) and contributes to maintenance of T cell stimulatory activity. AB - We reported previously that N-myc downstream-regulated gene 2 (NDRG2), a member of a new family of differentiation-related genes, is expressed specifically in dendritic cells (DC) differentiated from monocytes, CD34(+) progenitor cells, and the myelomonocytic leukemic cell line. In this study, we demonstrate that NDRG2 protein expression is detected, not only in in vitro-differentiated DC but also in primary DC from lymph nodes, thymus, and skin when anti-NDRG2 antibodies are used. As predicted from previous studies investigating the mRNA expression pattern of several types of cell lines, progenitor cells, and DC, NDRG2 protein was expressed strongly in DC. Its expression was detected at significant levels after differentiation from progenitor cells. RNA interference of NDRG2 demonstrated that activated leukocyte cell adhesion molecule (ALCAM) expression is down-regulated specifically in DC differentiated from NDRG2 small interfering RNA (siRNA)-transfected monocytes. This was consistent with our observation that U937 cells transfected with NDRG2 became resistant to the GM-CSF/IL-4-induced ALCAM reduction. Furthermore, DC, which had differentiated from NDRG2 siRNA transfected monocytes, showed a reduced ability to induce T cell proliferation. Taken together, our results indicate that NDRG2 is able to preserve ALCAM expression during DC differentiation from monocytes under cytokine culture conditions and that its expression helps DC maintain costimulatory signals necessary for T cell stimulation. PMID- 17911181 TI - Non-length dependent small fibre neuropathy/ganglionopathy. AB - OBJECTIVE: To describe the clinical and laboratory features of a painful non length dependent, small fibre ganglionopathy (SFG). BACKGROUND: The syndrome of generalised SFG with early involvement of the face, trunk or proximal limbs is not well recognised and contrasts with the burning feet syndrome of small fibre neuropathy (SFN) and classical large fibre features of sensory ganglionopathy. METHODS: Retrospective case review including skin biopsies from four neuromuscular centres. Patients with pre-existing diseases associated with ganglionopathies were excluded. RESULTS: 12 men and 11 women, with an average age of 50 years, were studied. Neuropathic pain developed over days in eight and over months in the other patients. The face (n = 12), scalp (n = 10), tongue (n = 6), trunk (n = 15) and acral extremities (n = 21) were involved. Symptoms began in the hands or face before the legs in 10. The pain was characterised as burning (n = 22), prickling (n = 13), shooting (n = 13) or allodynic (n = 11). There was loss of pinprick sensation in affected regions in 19, with minimal or no loss of large fibre sensibility. Laboratory findings included abnormal glucose metabolism in six patients, Sjogren syndrome in three and monoclonal gammopathy, sprue and hepatitis C infection in one each, with the remainder idiopathic. Sensory nerve action potentials were normal in 12 and were reduced in the hands but normal in the legs in six. Skin biopsy in 14 of 17 showed reduced nerve fibre density in the thigh equal to or more prominent than in the calf. Two of seven patients improved with immune therapies, 13 symptomatically with analgesic medications and the remainder had little improvement. Ten considered the pain disabling at the last follow-up (mean 2 years). CONCLUSION: The pattern of symmetric, non-length dependent neuropathic pain with face and trunk involvement suggests a selective disorder of the dorsal ganglia cells subserving small nerve fibres. It can be distinguished from distal SFN. A potential metabolic or immune process was detected in half of the cases and the disorder was often refractory to treatment. PMID- 17911182 TI - Effects of subthalamic deep brain stimulation on noun/verb generation and selection from competing alternatives in Parkinson's disease. AB - BACKGROUND AND AIMS: Impaired generation of verbs relative to nouns has been reported in Parkinson's disease (PD) and has been associated with the frontal pathophysiology of PD. The aim of the present study was to measure noun/verb generation abilities in PD and to determine whether noun/verb generation is affected by stimulation of the subthalamic nucleus (STN). PATIENTS AND METHODS: 8 participants who had been diagnosed with PD and had received surgery for deep brain stimulation (DBS) of the STN as well as 15 control participants completed a noun/verb generation task with four probe-response conditions-namely, noun-noun, verb-noun, noun-verb and verb-verb conditions. Patients with PD were assessed while receiving STN stimulation and without stimulation. RESULTS: During the off stimulation condition, patients with PD presented with a selective deficit in verb generation compared with control participants. However, when receiving STN stimulation, patients with PD produced significantly more errors than controls during the noun-noun and verb-verb conditions, supporting evidence from previous studies that STN stimulation modulates a frontotemporal network associated with word generation. Finally, errors during verb generation were significantly correlated with item selection constraint (ie, the degree to which a response competes with other response alternatives) in the on stimulation condition, but not the off stimulation condition. CONCLUSION: Our results suggest that STN stimulation affects the ability to select from many competing lexical alternatives during verb generation. PMID- 17911183 TI - Chlamydia pneumoniae seropositivity in aetiological subtypes of brain infarction and carotid atherosclerosis: a case control study. AB - BACKGROUND AND OBJECTIVE: Many patients with brain infarction (BI) lack traditional risk factors, suggesting that other factors (including infectious agents) might contribute to stroke risk. We investigated Chlamydia pneumoniae infection in a large cohort of patients with BI according to aetiological subtypes and carotid atherosclerosis. METHODS: We measured serum IgG and IgA to C. pneumoniae by microimmunofluorescence in 483 BI cases and 483 controls matched for age, sex and centre. IgG > or = 1/32 and IgA > or = 1/24 were considered positive. Cases with BI proven by magnetic resonance imaging were consecutively recruited and were classified into aetiological subtypes. Carotid atherosclerosis (intima-media thickness, plaques, stenosis) was evaluated by duplex ultrasonography in all subjects following the same method and with central reading. RESULTS: C. pneumoniae IgG seropositivity was not associated with BI (adjusted odds ratio (OR) 1.10, 95% confidence interval (CI) 0.80-1.51) and did not increase the risk of any aetiological subtype. Overall, C. pneumoniae IgA was not associated with BI (adjusted OR 1.54, 95% CI 0.84-2.81), but there was a significant interaction with hypertension. IgA seropositivity increased the BI risk in patients without hypertension (adjusted OR 2.79, 95% CI 1.15 to 6.74). When stratifying BI into subtypes, IgA seropositivity increased the risk of BI of unknown cause, but without significant heterogeneity. There was neither association with atherothrombotic, lacunar and cardioembolic BI nor with carotid intima-media thickness, carotid plaques or stenosis. CONCLUSIONS: We found no evidence that C. pneumoniae seropositivity is associated with carotid atherosclerosis and BI, regardless of aetiological subtype; but it might be associated with an increased risk of BI in normotensive patients. PMID- 17911184 TI - In vivo bioactivity of interferon-beta in multiple sclerosis patients with neutralising antibodies is titre-dependent. AB - BACKGROUND: Development of neutralising antibodies (NAbs) against recombinant interferon-beta (IFNbeta) is a significant clinical problem in the treatment of multiple sclerosis (MS). Several methods are available to assess NAbs, but there is a lack of consensus on how the different NAb titre levels interfere with the efficacy of the drug, especially in the individual patient. METHODS: NAb titres were measured with an in vitro MxA induction assay and the in vivo IFNbeta response was assessed by measuring MxA mRNA expression using real-time PCR. RESULTS: We identified titre levels of NAbs at which the IFNbeta biological activity was reduced or abrogated. Patients with NAb titres of up to 150 TRU/ml (ten times reduction units per ml) still had retained IFNbeta bioactivity, whereas greatly reduced levels of IFNbeta bioactivity were found in patients with NAbs of 150-600 TRU/ml. Titres above 600 TRU/ml were associated with loss of IFNbeta bioactivity. Similar results were obtained when TRAIL mRNA was used as a marker of the in vivo response to IFNbeta. CONCLUSION: There is a stepwise loss of IFNbeta bioactivity with increasing NAb titres and it is possible to identify functionally critical NAb titre levels that are useful to support treatment decisions at the individual patient level. PMID- 17911185 TI - Aceruloplasminaemia with progressive atrophy without brain iron overload: treatment with oral chelation. AB - BACKGROUND: Hereditary aceruloplasminaemia is a disorder of iron metabolism that is characterised by iron accumulation in the brain and other visceral organs. In previously reported cases, individuals with the disorder were noted to have evidence of iron accumulation in the brain. Oral chelating agents have not been used in neurological diseases of iron metabolism. METHODS: A 54-year-old woman who presented with ataxia, lower extremity spasticity and chorea was evaluated for evidence of the source of neurological dysfunction. RESULTS: Blood studies revealed no detectable ceruloplasmin. Marked iron overload was defined by a liver biopsy, which showed a variegated pattern consistent with a primary cause of iron overload. Review of MRI scans showed progressive brain atrophy without visible iron accumulation occurring over a 5-year period. The history suggested that neurodegeneration was coincident with aggressive oral iron replacement. Oral chelation improved many symptoms. CONCLUSIONS: Our findings in this patient suggest that disorders of iron transport such as aceruloplasminaemia can be a cause of neurological symptoms such as chorea and cognitive decline, as well as progressive neurodegeneration in the absence of visible iron on MRI scans. We found that oral iron chelation was effective at improving symptoms. PMID- 17911186 TI - Argatroban therapy for heparin-induced thrombocytopenia in acutely ill patients. AB - Heparin-induced thrombocytopenia (HIT) is a prothrombotic, immune-mediated adverse reaction to heparin therapy. To evaluate clinical outcomes and effects of argatroban therapy in acutely ill HIT patients. Retrospective analysis. Hospital in-patient. Acutely ill patients with clinically diagnosed HIT from previous multicenter, historically controlled studies of argatroban therapy in HIT. Argatroban, adjusted to maintain activated partial thromboplastin times 1.5 to 3 times baseline, or historical control therapy (ie, no direct thrombin inhibition). We identified 488 patients who received argatroban (N = 390; mean dose of 1.9 microg/kg/min for a mean 6 days) or historical control therapy (N = 98) for HIT. The primary all-cause composite endpoint of death, amputation, or new thrombosis within 37 days occurred in 133 (34.1%) argatroban-treated patients and 38 (38.8%) controls (P = .41). Argatroban, versus control, significantly reduced the primary thrombosis-related composite endpoint of death because of thrombosis, amputation secondary to ischemic complications of HIT, or new thrombosis (17.7% vs 30.6%, P = .007). Significant reductions also occurred in new thrombosis and death because of thrombosis. Major bleeding was similar between groups (7.7% vs 8.2%; P = .84). Adverse outcomes were more likely to occur in patients who were initially diagnosed with HIT and thrombosis, had undergone cardiac surgery, were not white, or had more severe thrombocytopenia. In acutely ill HIT patients, argatroban, versus historical control, provides effective antithrombotic therapy without increasing major bleeding. Patients with more severe thrombocytopenia or HIT-related thrombosis on HIT diagnosis have a poorer prognosis, emphasizing the importance of prompt recognition/ treatment of HIT in acutely ill patients. PMID- 17911187 TI - Plasma thrombin activatable fibrinolysis inhibitor and tissue factor pathway inhibitor changes following sepsis. AB - Sepsis-induced systemic inflammation results in coagulation abnormalities that may be different in gram-positive and gram-negative infections. We used ciprofloxacin to induce a predominantly gram-positive Enterococcus faecalis polymicrobial sepsis in rats. Ciprofloxacin-untreated rats exhibited a predominantly gram-negative polymicrobial sepsis. Rats were subjected to 30% body surface area burn (B), cecal ligation puncture (CLP) with a 22-gauge needle, and B + CLP. Ciprofloxacin-treated B + CLP rats showed a significant decrease in plasma thrombin activatable fibrinolysis inhibitor (TAFI) levels compared with sham rats. However, plasma tissue factor pathway inhibitor (TFPI) levels decreased significantly in B, CLP, and B + CLP groups compared with sham rats. The ciprofloxacin-untreated group showed a significant decrease in plasma TAFI levels in CLP and B + CLP and plasma TFPI levels decreased in all 3 groups compared with sham rats. Histological changes in the liver and kidney included vascular congestion and parenchyma bleed following B + CLP in ciprofloxacin untreated rats. These results suggest that plasma TAFI and TFPI levels differ depending on the type of bacteria involved in the septic process. PMID- 17911188 TI - Singlet oxygen enhances intrinsic thrombolysis: the intrinsic oxidative clot lysis assay (INOXCLA). AB - Granulocytes are important cells of inflammation and cellular thrombolysis. They produce urokinase (u-PA) and chloramines. In this study, u-PA/chloramine-mediated fibrinolysis is imitated in a microtiter-plate. Seventy-five microliters plasma are incubated with 50 microL 50% Pathromtin SL, 6% BSA, and 38 mM CaCl2 for 30 minutes (37 degrees C). Then, 50 microL 10 mM chloramine-T in PBS are added. After 30 minutes (37 degrees C), 50 microL 0, 100, or 10 IU/mL u-PA in 6% BSA-PBS are added and the turbidity is determined at 405 nm after 0, 3, or 16 hours. Clot lysis was increased more than tenfold by 0.5 to 1 micromoles chloramine (ED50 after 3h = about 0.25 micromoles = 2 mM final concentration). The normal range for the present intrinsic oxidative clot lysis assay (INOXCLA) is 100% +/- 25% (MV +/- SD; 100 relative % of norm; the normal lysis being 60 absolute %; CVs < 10%). Fifty percent lysis of adherent microclots occurred after 0.75 hours, 2 hours, 14 hours, 13 days, or 17 days when using 1000, 100, 10, 1, or 0 IU/mL u-PA reagent. If the u-PA activity is quenched by PAI-2, no clot lysis appears. Chloramines are important physiologic generators of nonradical excited singlet oxygen and enhance u-PA-mediated lysis of plasma clots. Based on the u PA/chloramines coaction, a new global fibrinolysis assay has been derived. PMID- 17911189 TI - Type IIb von Willebrand disease: role of qualitative defects in atherosclerosis and endothelial dysfunction. AB - OBJECTIVE: To verify whether a hereditary bleeding tendency, such as von Willebrand disease (vWD) type IIB, protects against the onset of atherosclerosis. PARTICIPANTS AND METHODS: Twenty-four patients with vWD type IIB and 24 healthy controls, matched for common atherosclerotic risk factors. All patients were evaluated by color Doppler ultrasound of the common carotid, carotid bifurcation, common femoral artery, brachial artery, and abdominal aorta, investigating intima media thickness (IMT) and presence of plaques in each arterial district. Flow mediated dilation (FMD) of the brachial artery was used to test endothelial function. RESULTS: vWD type IIB patients presented no significant difference in IMT in any arterial district. FMD showed no differences between the 2 groups. CONCLUSIONS: The quantitative clotting defect characteristic of vWD type IIB does not seem to protect against atherosclerosis. PMID- 17911190 TI - Elevated concentrations of soluble adhesion molecules and large platelets in diabetic patients: are they markers of vascular disease and diabetic nephropathy? AB - P-selectin, E-selectin, and mean platelet volume are markers associated with platelet reactivity that have been demonstrated to be increased in diabetes. We were particularly interested to see if there was a difference in mean platelet volume and selectins between diabetics and nondiabetics, and in diabetics with and without nephropathy, and whether there was a correlation between mean platelet volume and selectins. One hundred and fourteen diabetic patients and 31 healthy controls were investigated. Plasma levels of P-selectin and E-selectin were higher in the diabetic group than in controls (P = .001 and P = .007, respectively) and in diabetic patients with proteinuria than in patients without proteinuria (P = .002 and P = .004, respectively). Protein excretion was lower in patients with low mean platelet volume values (P = .004). In conclusion, elevated platelet volume and high selectin values may play a role in the development of vasculopathies and complications in diabetes mellitus. Further studies are needed to prove these results. PMID- 17911191 TI - Hyperhomocysteinemia relates to the subtype of antiphospholipid antibodies in non SLE patients. AB - Abnormal increases of antiphospholipid antibody and plasma homocysteine levels are recently emerging as nonlipidic risk factors for cerebral atherogenesis and thrombosis. Both antiphospholipid antibody and homocysteine share many similar bioeffects in hemostasis, but their interaction is still inconsistent. In this study, we examined the relation between the plasma homocysteine level and lupus anticoagulant, anticardiolipin antibody, and anti-beta2-glycoprotein I antibody in patients with noncardiac cerebral ischemia. Systemic lupus erythrematosus patients were excluded. The results showed a higher frequency of moderate hyperhomocysteinemia in patients with an abnormal increase of lupus anticoagulant only. Neither the serum folate and cobalamin levels nor methylenetetrahydrofolate reductase allele mutation contributes to this result. Accordingly, homocysteine interacts with lupus anticoagulant to promote cerebral atherosclerosis and ischemia. The role of vasculopathic or prothrombotic autoantibody generation in response to specific pathological change such as hyperhomocysteinemia warrants further investigation. PMID- 17911192 TI - Variability in anticardiolipin antibody detection: role of nonspecific IgG binding and different microtiter plates. AB - There are many studies that are available on the Internet that attempt to standardize the assay for anticardiolipin antibody evaluation because of the variability of results. The aim of this study was to evaluate simultaneously the role of different microplates and the importance of sample nonspecific binding in determining different results in anticardiolipin antibody detection. Sera from 8 patients with raised levels of IgG anticardiolipin antibodies and 10 control sera were assayed by enzyme-linked immunosorbent assay in the presence (specific binding) or in the absence of cardiolipin (sample blank) with four different microplates, that is, NUNC PolySorp, FALCON ProBIND, Greiner 655061 (high binding), and Greiner 655001 (medium binding). Results were expressed as optical densities or net-optical densities (following sample blank subtraction) as well as international IgG anticardiolipin units (GPL) or net-GPL. A wide interplate variability of optical densities was found. When results were expressed as GPL, significant differences were only found between Greiner 655061, FALCON ProBIND, and NUNC PolySorp (P < .05 and P < .001, respectively) whereas differences were not statistically significant if interplate variability was analyzed as net-GPL. Results expressed as categorical variables (ie, positive/negative, according to a GPL cut-off and net-GPL cut-off, obtained with sera from 100 apparently healthy blood donors) showed a good or excellent Cohen's kappa coefficient of concordance among plates when positivity was evaluated on net-GPL. Our data strongly suggest that quantification and subtraction of sample blank may improve both interlaboratory agreement and reliability of anticardiolipin assay and minimize false-positive results. PMID- 17911193 TI - Soluble P-selectin during a single hemodialysis session in patients with chronic renal failure and erythropoietin treatment. AB - In several studies, hemodialysis (HD) patients treated with recombinant human erythropoietin (rHuEPO) because of renal anemia showed increased levels of soluble adhesion molecules. The purpose of the study was to investigate the changes of soluble P-selectin (sSELP) and its relationship to platelet activation during a single HD session in patients with long-term rHuEPO treatment. Fifty-two HD patients with chronic renal failure were involved--26 with rHuEPO treatment (EPO group) and 26 without (non-EPO group). Thirty healthy subjects served as the control group. The sSELP, beta-thromboglobulin, and platelet factor 4 plasma levels were measured before and after a single 4-hour HD session on a cuprophane dialyzer. The basal beta-thromboglobulin and platelet factor 4 plasma levels were significantly increased in both HD groups compared with healthy controls but did not change after a single HD session, except for a significant decrease of platelet factor 4 in the non-EPO group. The predialysis sSELP plasma levels did not differ significantly compared with those of the healthy controls, but there was a significant increase of sSELP levels after a single HD session in both groups (EPO, P < .005; non-EPO, P < .05, respectively). These results suppose that the increased sSELP level was released from platelets during the course of a single HD session. The more significant increase of the sSELP plasma levels in EPO group during HD indicates that platelets are more activated in patients with long-term rHuEPO treatment, and this fact could partially explain the suspected tendency for thrombosis in these patients. PMID- 17911194 TI - Influence of radiofrequency catheter ablation on platelet aggregation. AB - The objective of this article was to find out how radiofrequency catheter ablation (RFA) influences platelet aggregation (PA), and the dependence on the total energy (TE) of RFA used and the cause of arrhythmia. We investigated 97 patients. PA was analyzed before, after, and in 24 hours after RFA. ADP- and epinephrine-induced PA significantly decreased after RFA by 5% and 8.9% (P < .001), respectively, and increased in 24 hours close to baseline. PA induced by ADP and collagen did not radically depend on the TE. Epinephrine-induced PA decreased after RFA by 0%, 8% (P < .05), and 16.9% (P < .01) in groups of patients where the TEs used were <4000 J, 4000 to 15,000 J, and >15,000 J, respectively. There were no significant differences in PA between groups based on the cause of arrhythmia. ADP- and epinephrine-induced PA significantly decreased after RFA and returned close to baseline in 24 hours. Epinephrine-induced PA was inversely associated with the TE used for RFA. PMID- 17911195 TI - Hypercoagulable state in five thalassemia intermedia patients. AB - Fifty-three patients of thalassemia intermedia and 40 controls were studied for clinical evidence of thrombosis and laboratory evidence of hypercoagulable state. Thrombotic episodes were detected in 5 (9.4%) patients. Two of these 5 patients with thrombosis were splenectomized. Laboratory evaluation showed presence of thrombocytosis in 8 (15%), 5 of these were splenectomized. Platelet hyperaggregation was detected in 12 (22.2%) patients. Although rate of aggregation was slow in 7 (13.2%) patients, degree of aggregation was normal in these 7 patients and platelet hypoaggregation was not detected in any patient. Level of coagulation inhibitors protein C and protein S, and antithrombin III were decreased in 31 (58.4%) patients. There was no correlation between low level of protein C and protein S with hepatic dysfunction and iron overload. Antithrombin III level was decreased only in 8 (15%) patients. There was a statistically significant association between the lower level of this inhibitor and hepatic dysfunction. In conclusion, this study provides evidence for the existence of a chronic hypercoagulable state in patients with beta thalassemia intermedia, and suggests that expression of a procoagulant surface by thalassemia intermedia red blood cells may be the major underlying factor giving rise to platelet and coagulation inhibitor abnormalities in these patients. These alterations are not related to iron overload or hepatic dysfunction. PMID- 17911196 TI - Evaluation of platelet and leukocyte functions in effort angina patients using high shear conditions in small-sized collagen bead columns. AB - This study used a small-sized collagen bead column system to investigate platelet functions under high shear stress in 28 patients with effort angina and 15 healthy controls. Soluble P-selectin and soluble L-selectin were also evaluated. Patients underwent stent insertion, followed by treatment with aspirin (100 mg/day) and ticlopidine (200 mg/day). High-shear-dependent platelet function was measured using small-sized collagen beads. The retention rate of platelets from patients with angina was higher than that in healthy controls (10.9% +/- 3.9% versus 5.6% +/- 1.7%, P < .01). Soluble P-selectin and soluble L-selectin levels of patients with angina significantly increased after passage through the columns. Levels of soluble P-selectin and L-selectin in healthy controls did not exhibit significant changes with passage through the columns. These results suggested that high shear stress caused abnormal activation of leukocytes or adhesive proteins in patients with effort angina, and ticlopidine failed to suppress hyperaggregability of platelets in these patients. PMID- 17911197 TI - Molecular analysis of factor V Leiden, factor V Hong Kong, factor II G20210A, methylenetetrahydrofolate reductase C677T, and A1298C mutations related to Turkish thrombosis patients. AB - Inherited gene disorders related to the hemostatic system have been documented as risk factors for thrombosis. The roles of factor V Hong Kong (FV Hong Kong), factor V Leiden (FV Leiden), factor II G20210A (FII G20210A), methylenetetrahydrofolate reductase (MTHFR) C677T, and MTHFR A1298C mutations in Turkish patients with thrombosis (270 patients) compared with healthy controls (114 subjects) were evaluated. Polymerase chain reaction-based restriction enzyme analysis was carried out to screen these mutations, and single-strand conformation analysis was established to identify variations using the primers selected for restriction enzyme analysis studies. As a result, a significant relationship was determined among FV Leiden, FII G20210A, and thrombosis. The FV Hong Kong mutation was observed in only 2 patients with pulmonary vein thrombosis who are FV Leiden/FV Hong Kong compound heterozygous for FV gene. MTHFR C677T and A1298C were equally distributed in the patient group compared with the control group. All named mutations were also identified with single-strand conformation analysis, but a new variant/polymorphism during studies was not found. Because some inherited abnormalities are associated with thromboembolic disorders, determining the mutations and gene-to-gene interactions in patients with thrombosis history has a great impact on diagnosis and treatment of these diseases. PMID- 17911198 TI - Acute anterior myocardial infarction after chemotherapy for testicular seminoma in a young patient. AB - Testicular cancer is the most common solid tumor among young men aged 15 to 35 years. Combination chemotherapy with cisplatin, etoposide, and bleomycin remains the mainstay of treatment. We present a 27-year-old man who presented with an acute anterior myocardial infarction during the second course of chemotherapy for seminoma. Because the patient had no significant risk factors for coronary heart disease, the infarction was likely caused by the chemotherapy regimen. PMID- 17911199 TI - Acquired hemophilia: a case report of 2 patients with acquired factor VIII inhibitor treated with rituximab plus a short course of steroid and review of the literature. AB - Acquired hemophilia is an unusual disorder in which nonhemophiliac patients develop autoantibodies (inhibitor) against the factor VIII coagulation protein. Factor VIII inhibitor leads to life-threatening bleeding disorders classically described as new onset of diffuse bruising and prolonged partial thromboplastin time in elderly patients. Treatment is focused in the control of the acute bleeding episode and the long-term suppression of the autoantibody. Several immunosuppressive combinations have been described; however, these treatments are also associated with serious side effects that are difficult to tolerate, especially in older and debilitated patients. New treatment modalities explore the elimination of the autoantibody production by targeting B-cells with rituximab, an anti CD-20 monoclonal antibody that has shown success in a multitude of autoimmune processes. This report presents 2 patients successfully treated with rituximab and a short tapering course of steroids and focuses our discussion in the analysis of different treatment approaches available for these patients' population. PMID- 17911200 TI - Thrombotic thrombocytopenic purpura and bone marrow necrosis as the initial presentation of lung cancer. AB - Thrombotic thrombocytopenic purpura (TTP) is a disseminated form of thrombotic microangiopathy. Although most cases are held to be idiopathic, its association with malignancy is well recognized, and it usually occurs at the terminal stage of cancer. Bone marrow necrosis (BMN) is another rare disorder defined pathologically as the necrosis of myeloid tissue and medullary stroma with preservation of bone. Although hematologic malignancy is the most common underlying disease associated with BMN, it can also be caused by solid tumors. The occurrence of TTP with BMN associated with lung cancer has not been reported in the English literature. First described is a patient with the rare association of TTP and BMN displayed as the first manifestations of a lung cancer. PMID- 17911201 TI - Enoxaparin-induced reversible pancytopenia. PMID- 17911202 TI - Inherited warfarin resistance in Indian patients: does it occur? PMID- 17911203 TI - Safety concerns with fluoroquinolones. AB - OBJECTIVE: To review the chemistry, pharmacology, and safety of fluoroquinolones. DATA SOURCES: A MEDLINE search (1966-July 2007) was conducted using the key words fluoroquinolones or quinolones with safety, adverse effects, hypoglycemia, hyperglycemia, dysglycemia, QTc prolongation, torsades, seizures, phototoxicity, tendon rupture, Clostridium difficile, and pseudomembranous colitis for articles published in the English language. STUDY SELECTION AND DATA EXTRACTION: Medicinal chemistry, in vitro, animal, and human trials were reviewed for information on the chemistry, pharmacology, and safety of each fluoroquinolone. Clinical trials were reviewed and included to compare the safety of systemic fluoroquinolones on the market. Literature on the pathology of serious adverse effects was also reviewed. DATA SYNTHESIS: Gatifloxacin has been shown to increase the risk of hospitalization for dysglycemia in patients with and without diabetes. Hyperglycemia may occur with any fluoroquinolone, especially if not properly dose adjusted. Hypoglycemia may occur with any fluoroquinolone and has a higher frequency in patients receiving concomitant oral hypoglycemic drugs or insulin. Use of any fluoroquinolone should be avoided in patients with risk factors for QTc interval prolongation or tendinopathy. All fluoroquinolones should be used with caution in patients with a history of seizure disorders and may cause phototoxicity or C. difficile-associated diarrhea (CDAD). CONCLUSIONS: Clinicians should be aware of possible alterations in blood glucose, QTc interval prolongation, seizures, phototoxicity, tendinopathy, or CDAD with the use of any fluoroquinolone, especially in patients with other risk factors for these conditions. Clinicians should closely monitor for these adverse effects and appropriately adjust doses to minimize these risks. To provide safe treatment for patients needing antibiotic therapy, an assessment of the risk-benefit ratio may be warranted in the decision to use a fluoroquinolone. PMID- 17911204 TI - Efficacy and tolerability of topiramate in vascular generalized chorea. PMID- 17911205 TI - Ursodiol in patients with parenteral nutrition-associated cholestasis. AB - OBJECTIVE: To review the role of ursodeoxycholic acid (ursodiol) in treating parenteral nutrition-associated cholestasis (PNAC). DATA SOURCES: A MEDLINE (1950 May 2007) search was performed using the key terms parenteral nutrition, cholestasis, ursodeoxycholic acid, and ursodiol. STUDY SELECTION AND DATA EXTRACTION: All English-language articles that evaluated the safety and efficacy of ursodiol for PNAC were included in this review. DATA SYNTHESIS: The benefits of exogenous ursodiol administration in the treatment of cholestasis can be explained by its alteration of effects on bile composition and flow and provision of cytoprotective, membrane stabilizing, and immunomodulatory effects. Two animal studies, 2 case reports, and 6 human studies (2 prospective and 3 retrospective pediatric studies, 1 adult prospective study) evaluated the efficacy of ursodiol in patients with PNAC. Ursodiol 10-30 mg/kg/day in children and 10-15 mg/kg/day in adults administered in 2-3 doses improved the biochemical and clinical signs and symptoms of PNAC. However, short-term improvement in biochemical parameters may not necessarily predict the outcome of PNAC patients. At recommended doses, ursodiol may not be effective in patients with short bowel syndrome or in those with resected terminal ileum because of reduced ursodiol absorption. Studies supporting the efficacy of ursodiol in treatment of PNAC are limited by small sample size, absence of randomization and controls, short duration, and lack of accountancy to confounding variables. Large, prospective, randomized, placebo controlled, long-term follow-up studies evaluating the efficacy and optimal dosing and duration of ursodiol therapy for PNAC are not yet available. CONCLUSIONS: Ursodiol may improve the biochemical signs and clinical symptoms of PNAC. However, optimal dosing, timing, duration of therapy, and long-term effects on PNAC outcome and prognosis require further studies. PMID- 17911207 TI - Comment: Prazosin treatment of nightmares related to posttraumatic stress disorder. PMID- 17911206 TI - Optimal initial dose adjustment of warfarin in orthopedic patients. AB - BACKGROUND: Warfarin sodium is commonly prescribed for the prophylaxis and treatment of venous thromboembolism. Dosing algorithms have not been widely adopted because they require a fixed initial warfarin dose (eg, 5 mg) and are not tailored to other factors that may affect the international normalized ratio (INR). OBJECTIVE: To develop an algorithm that could predict a therapeutic warfarin dose based on drug interactions, INR response after the initial warfarin doses, and other clinical factors. METHODS: We used stepwise regression to quantify the relationship between these factors in patients beginning prophylactic warfarin therapy immediately prior to joint replacement. In the derivation cohort (n = 271), we separately modeled the therapeutic dose after 2 and 3 initial doses. We prospectively validated these 2 models in an independent cohort (n = 105). RESULTS: About half of the therapeutic dose variability was predictable after 3 days of therapy: R2 was 53% in the derivation cohort and 42% in the validation cohort. INR response after 3 warfarin doses (INR3) inversely correlated with therapeutic dose (p < 0.001). Intraoperative blood loss transiently, but significantly, elevated the postoperative INR values. Other significant (p < 0.03) predictors were the first and second warfarin doses (+7% and +6%, respectively, per 1 mg), and statin use (-15.0%). The model derived after 2 warfarin doses explained 32% of the variability in therapeutic dose. CONCLUSIONS: We developed and validated algorithms that estimate therapeutic warfarin doses based on clinical factors and INR response available after 2-3 days of warfarin therapy. The algorithms are implemented online at www.WarfarinDosing.org. PMID- 17911209 TI - Comment: Epidemiology, risk factors, and outcomes of Candida albicans versus non albicans candidemia in nonneutropenic patients. PMID- 17911210 TI - Watching others' actions: mirror representations in the parietal cortex. AB - An observation that neurons in the motor cortex of the monkey are active both when the monkey performs a specific action and when he watches an actor executing the same action led to the mirror-system hypothesis. This hypothesis suggests that primates perceive and interpret others' actions by generating an internal motor representation (e.g., simulation). Recent evidence suggests that humans have a similar mirror system. In this review, we focus on the essential congruence between the motor and visual properties of an action. We summarize behavioral and imaging studies in humans that show that observing others' actions can interfere with our own motor execution. We discuss a framework for understanding such an internal representation and suggest that the activity in the parietal cortex during observation of others' actions is based on the sensory to-motor remapping properties of this region, which are necessary for fine control of our own actions. PMID- 17911211 TI - Neurogenesis in the cerebellum. AB - In the past few years, genetic fate mapping experiments have changed our vision of cerebellar development, particularly in redefining the origin of gabaergic and glutamatergic neurons of the cerebellar cortex and highlighting the precise spatio-temporal sequence of their generation. Here the authors review cerebellar neurogenesis and discuss the fate mapping studies with other new information stemming from transplantation experiments, in an effort to link the developmental potential of neural progenitor populations of the cerebellum with their spatio temporal origin. PMID- 17911212 TI - Cerebral disconnectivity: an early event in schizophrenia. AB - Neuroimaging and electrophysiological investigations have demonstrated numerous differences in brain morphology and function of chronic schizophrenia patients compared to healthy controls. Studying patients at the beginning of their disease without the confounding effects of chronicity, medication, and institutionalization may provide a better understanding of schizophrenia. Recently, at many institutions around the world, special projects have been launched for specialized treatment and research of this interesting patient group. Using the PubMed search engine in this update, the authors summarize recent investigations between January 2002 and September 2006 that focus on whether signs of disconnectivity already exist early in the disease process. They discuss gray and white matter changes, their impact on symptomatology, electroencephalogram-based studies on connectivity, and possible influences of medication. PMID- 17911213 TI - The endoplasmic reticulum as an integrator of multiple dendritic events. AB - Dendrites are integrating elements that receive numerous subsets of heterogeneous synaptic inputs, which generate temporally and spatially distinct changes in membrane potential and intracellular Ca2+ levels in local domains. The ubiquitously distributed endoplasmic reticulum (ER) in dendrites is luminally connected to the bulk ER in the soma, constituting a huge interconnected intracellular network that allows rapid Ca2+ diffusion and equilibration. The ER is an excitable organelle that can elicit or terminate cytosolic Ca2+ signals in local or global domains. The absolute level or changes in the Ca2+ concentration in the ER lumen are also very important for the synthesis and maturation of proteins, regulation of gene expression, mitochondrial functions, neuronal excitability, and synaptic plasticity. Through the connected lumen of the ER, information from multiple dendritic events in neurons appears to be delivered into the bulk ER in the soma. Therefore, the ER network in neurons is emerging as a conveyor and integrator of signals. In this article, we will discuss the various roles of the ER and the functional and structural organization of the ER network in neurons. PMID- 17911214 TI - Amyloid precursor protein and mitochondrial dysfunction in Alzheimer's disease. AB - Growing evidence suggests that mitochondrial dysfunction is one of the key intracellular lesions associated with the pathogenesis of Alzheimer's disease (AD). Mitochondria, the powerhouses of the cell, participate in a number of physiological functions that include calcium homeostasis, signal transduction, and apoptosis. However, the pathophysiological mechanisms underlying the decline of mitochondrial vital functions leading to the dysfunction of mitochondria during AD are not well understood. Recent literature has observed the accumulation of Alzheimer's amyloid precursor protein (APP) and its C-terminal cleaved product beta-amyloid (Abeta) in the mitochondrial compartment. Furthermore, evidence also implicates that the accumulation of full-length APP and Abeta in the mitochondrial compartment has a causative role in impairing mitochondrial physiological functions. Here, we review the mode of mitochondrial transport of full-length APP and Abeta and its pathological implications in bringing about mitochondrial dysfunction as seen in AD. PMID- 17911215 TI - Functional anatomic models of language: assembling the pieces. AB - In the past few years, a series of influential review articles have summarized the state of the art with respect to cortical models of language organization. The present article is a mini-review and conceptual meta-analysis of several of the most prominent recent contributions. Based on the models, the authors extract some generalizations to arrive at a more robust model that 1) does justice to the range of neurological data, 2) is more connected to research in linguistics and psycholinguistics, and 3) stimulates hypothesis-driven research in this domain. In particular, the article attempts to unify a few of the current large-scale models of the functional neuroanatomy of language in a more principled manner. First, the authors argue that the relevant type of processing in a given cortical area, that is, memorizing (temporal cortex) versus analyzing (parietal) versus synthesizing (frontal), lies at the basis of local neuronal structure and function. Second, from an anatomic perspective, more dorsal regions within each of these (temporal, parietal, and frontal) systems specialize more in phonological processing, middle areas in syntactic processing, and more ventral areas in semantic processing. PMID- 17911216 TI - The role of the right temporoparietal junction in social interaction: how low level computational processes contribute to meta-cognition. AB - Accumulating evidence from cognitive neuroscience indicates that the right inferior parietal cortex, at the junction with the posterior temporal cortex, plays a critical role in various aspects of social cognition such as theory of mind and empathy. With a quantitative meta-analysis of 70 functional neuroimaging studies, the authors demonstrate that this area is also engaged in lower-level (bottom-up) computational processes associated with the sense of agency and reorienting attention to salient stimuli. It is argued that this domain-general computational mechanism is crucial for higher level social cognitive processing. PMID- 17911218 TI - Neuron glial communication at synapses: insights from vertebrates and invertebrates. AB - Glial cells are instrumental for many aspects of nervous-system function. Interestingly, complex neuron-glial interactions at synapses are commonly found in both invertebrates and vertebrates. Although these interactions are known to be important for synaptic physiology, the cellular processes and molecular mechanisms involved have not been fully uncovered. Identifying the common and unique features of neuron-glial interactions between invertebrates and vertebrates may provide valuable insights into the relationship of neuron-glial cross-talk to nervous-system function. This review highlights selected studies that have revealed structural and functional insights into neuron-glial interactions at synapses in invertebrate and vertebrate model systems. PMID- 17911219 TI - BDNF and memory formation and storage. AB - During the past decade, a large body of evidence has implicated BDNF in synaptic plasticity. In this review, we focus on the newer experiments that involve BDNF in different aspects of learning and memory processing-in particular, in memory persistence and storage. PMID- 17911220 TI - Mitochondrial dynamics and peripheral neuropathy. AB - Peripheral neuropathy is perhaps the archetypal disease of axonal degeneration, characteristically involving degeneration of the longest axons in the body. Evidence from both inherited and acquired forms of peripheral neuropathy strongly supports that the primary pathology is in the axons themselves and points to disruption of axonal transport as an important disease mechanism. Recent studies in human genetics have further identified abnormalities in mitochondrial dynamics -the fusion, fission, and movement of mitochondria--as a player in the pathogenesis of inherited peripheral neuropathy. This review provides an update on the mechanisms of mitochondrial trafficking in axons and the emerging relationship between the disruption of mitochondrial dynamics and axonal degeneration. Evidence suggests mitochondria are a "critical cargo" whose transport is necessary for proper axonal and synaptic function. Importantly, understanding the regulation of mitochondrial movement and the consequences of decreased axonal mitochondrial function may define new paths for therapeutic agents in peripheral neuropathy and other neurodegenerative diseases. PMID- 17911221 TI - On the hypes and falls in neuroprotection: targeting the NMDA receptor. AB - Activation of the NMDA (N-methyl-D-aspartate) responsive subclass of glutamate receptors is an important mechanism of excitatory synaptic transmission. Moreover, NMDA receptors are widely involved in many forms of synaptic plasticity such as long-term potentiation (LTP) and long-term depression (LTD), which are thought to underlie complex tasks, including learning and memory. Dysfunction of these ligand-gated cation channels has been identified as an underlying molecular mechanism in neurological disorders ranging from acute stroke to chronic neurodegeneration in amyotrophic lateral sclerosis. Excessive glutamate levels have been detected following brain trauma and cerebral ischemia, resulting in an unregulated stimulation of NMDA receptors. These conditions are thought to elicit a cascade of excitation-mediated neuronal damage where massive increases in intracellular calcium concentrations finally trigger neuronal damage and apoptosis. Consistent with the hypothesis of NMDA receptors as essential mediators of excitotoxicity, the different functional domains of these ion channels have been identified as potential targets for neuroprotective agents. Following an initial hype on potential NMDA receptor therapeutics, the authors currently see a period of skepticism that, in reverse, appears to neglect the therapeutic potential of this receptor class. This review attempts a reappraisal of this important class of neurotransmitter receptors, with a focus on NMDA receptor heterogeneity, ligand binding domains, and candidate diseases for a potential neuroprotective therapy. PMID- 17911222 TI - . . . And the olive said to the cerebellum: organization and functional significance of the olivo-cerebellar system. AB - What is the role of the olive in cerebellar operations? And what is the role of the cerebellum in the sensory and motor functioning of the nervous system? It is clear that the olivo-cerebellar network plays a key role in the managing of vertebrate motor control, as lesions in this network cause motor deficits in humans and animals. However, it is increasingly becoming clear that the olivo cerebellar system is involved in the control of more than simple motor behaviors. The elegant and almost geometric organization of the olivocerebellar network lends itself to performing these complex operations. In this review, the salient anatomical, physiological, and clinical features of this system are discussed. A computer-assisted visualization system is used to illustrate some of the anatomical points. The idea that the cerebellum and the olivo-cerebellar networks are perhaps in the upper echelons of the hierarchy of brain functioning, if such a hierarchy indeed exists, is discussed. PMID- 17911223 TI - Similarity and diversity in visual cortex: is there a unifying theory of cortical computation? AB - The cerebral cortex, with its conserved 6-layer structure, has inspired many unifying models of function. However, recent comparative studies of primary visual cortex have revealed considerable structural diversity, raising doubts about the possibility of an all-encompassing theory. This review examines similarities and differences in V1 across mammals. Gross laminar interconnections are relatively conserved. Major functional response classes are found universally or nearly universally. Orientation and spatial frequency tuning bandwidths are quite similar despite an enormous range of visual resolution across species, and orientation tuning is contrast-invariant. Nevertheless, there is considerable diversity in the abundance of different cell classes, laminar organization, functional architecture, and functional connectivity. Orientation-selective responses arise in different layers in different species. Some mammals have elaborate columnar architecture like orientation maps and ocular dominance bands, but others lack this organization with no apparent impact on single cell properties. Finally, local functional connectivity varies according to map structure: similar cells are connected in smooth map regions but dissimilar cells are linked in animals without maps. If there is a single structure/function relation for cortex, it must accommodate significant variations in cortical circuitry. Alternatively, natural selection may craft unique circuits that function differently in each species. PMID- 17911225 TI - Outcomes from studies of antineutrophil cytoplasm antibody associated vasculitis: a systematic review by the European League Against Rheumatism systemic vasculitis task force. AB - OBJECTIVES: We undertook a systematic literature review as a background to the European League Against Rheumatism (EULAR) recommendations for conducting clinical trials in anti-neutrophil cytoplasm antibody associated vasculitis (AAV), and to assess the quality of evidence for outcome measures in AAV. METHODS: Using a systematic Medline search, we categorised the identified studies according to diagnoses. Factors affecting remission, relapse, renal function and overall survival were identified. RESULTS: A total of 44 papers were reviewed from 502 identified by our search criteria. There was considerable inconsistency in definitions of end points. Remission rates varied from 30% to 93% in Wegener granulomatosis (WG), 75% to 89% in microscopic polyangiitis (MPA) and 81% to 91% in Churg-Strauss syndrome (CSS). The 5-year survival for WG, MPA and CSS was 74 91%, 45-76% and 60-97%. Relapse (variably defined) was common in the first 2 years but the frequency varied: 18% to 60% in WG, 8% in MPA, and 35% in CSS. The rate of renal survival in WG varied from 23% at 15 months to 23% at 120 months. METHOD: used to assess morbidity varied between studies. Ignoring the variations in definitions of the stage of disease, factors influencing remission, relapse, renal and overall survival included immunosuppressive therapy used, type of organ involvement, presence of ANCA, older age and male gender. CONCLUSIONS: Factors influencing remission, relapse, renal and overall survival include the type of immunosuppressive therapy used, pattern of organ involvement, presence of ANCA, older age and male gender. Methodological variations between studies highlight the need for a consensus on terminology and definitions for future conduct of clinical studies in AAV. PMID- 17911224 TI - TNFalpha: a trigger of autonomic dysfunction. AB - During disease, infection, or trauma, the cytokine tumor necrosis factor alpha (TNF alpha) causes fever, fatigue, malaise, allodynia, anorexia, gastric stasis associated with nausea, and emesis via interactions with the central nervous system. Our studies have focused on how TNF alpha produces a profound gastric stasis by acting on vago-vagal reflex circuits in the brainstem. Sensory elements of this circuit (i.e., nucleus of the solitary tract [NST] and area postrema) are activated by TNF alpha. In response, the efferent elements (i.e., dorsal motor neurons of the vagus) cause gastroinhibition via their action on the gastric enteric plexus. We find that TNF alpha presynaptically modulates the release of glutamate from primary vagal afferents to the NST and can amplify vagal afferent responsiveness by sensitizing presynaptic intracellular calcium-release mechanisms. The constitutive presence of TNF alpha receptors on these afferents and their ability to amplify afferent signals may explain how TNF alpha can completely disrupt autonomic control of the gut. PMID- 17911226 TI - Predicting beef tenderness using near-infrared spectroscopy. AB - The objective of this multiple-phase study was to determine the accuracy of an on line near-infrared (NIR) spectral reflectance system to predict 14-d-aged cooked beef tenderness. In phase I, 292 carcasses (140 US Select, 152 US Choice) were selected (d 2) from 2 commercial beef processing facilities. After carcass selection, longissimus lumborum (LL) muscle sections (ribs 9 to 12) were individually identified, vacuum-packaged, and transported to the Oklahoma State University Meats Laboratory, where a 2.54-cm-thick steak (n = 1) was fabricated and stored in refrigerated conditions (1 degrees C +/- 1). Following a 30-min oxygenation period, a NIR spectral scan was obtained on the 12th-rib LL steak. Steaks (d 3) were individually vacuum-packaged and aged at 4 degrees C for a total of 14 d before cooking slice shear force (SSF) analysis. In phases II and III, 476 carcasses (258 US Select, 218 US Choice) were immediately NIR scanned after carcass presentation to in-plant USDA grading personnel. In a similar fashion, all LL steaks were aged (1 degrees C +/- 1) for 14 d before cooking (70 degrees C) and conducting SSF. Of the phase I and II samples, 39 (6.77%) were categorized as being tough (i.e., >/= 25 kg of SSF after the 14-d postmortem aging period). Of these 39 tough samples, 20 (3.7% error rate) were correctly placed in the 90% certification level. Another 10 tough samples were placed in the 80% certification level (2.0% error rate). The overall NIR certified tender group was 1.67 kg more tender (P < 0.05) than LL samples from the noncertified samples. When the NIR predicted samples to be tough, 10% of the samples were eliminated from the phase I and II LL populations at 90% certification. The population SSF mean improved in excess of 6.5 kg. For phase III, SSF evaluation by an independent third party indicated the NIR system was able to successfully sort tough from tender LL samples to 70% certification levels. It was concluded that NIR scanning offers an in-plant opportunity to sort carcasses into tenderness outcome groups for guaranteed-tender branded beef programs. PMID- 17911227 TI - Salmonella challenges: prevalence in swine and poultry and potential pathogenicity of such isolates. AB - Salmonellosis is the second leading cause of bacterial foodborne illness in the United States, and the great majority of these infections are associated with the consumption of products such as meat, poultry, eggs, milk, seafood, and fresh produce contaminated with Salmonella. The per capita consumption of meat and poultry in United States has increased significantly over the past century. This increase is especially evident with poultry products, where there has been a nearly 6-fold increase in chicken consumption and 17-fold increase in turkey consumption since 1909. The per capita consumption of pork has also increased over this time from 18.7 to 21.7 kg/yr. With this increase in meat and poultry consumption, the dynamics of animal production and consumer exposure have changed leading to new challenges in limiting salmonellosis. To meet the demands of consumers, more intensive agricultural practices have been adopted, which has likely changed the population characteristics of Salmonella present among poultry flocks and swine populations. In Salmonella isolated from swine in the United States, S. Typhimurium has replaced S. Choleraesuis as the predominant serovar in recent years. Among isolates from turkeys collected in 2004, serovars S. Senftenberg and S. Hadar were most common overall; however, S. Heidelberg was most common from clinical diagnostic sources, potentially indicating increased virulence. Salmonella Heidelberg was also the most commonly detected serovar among chicken isolates from clinically ill birds and Salmonella surveillance samples. Overall among the 10 serovars most commonly associated with human infections, 6 are also found in the top serovars of swine and poultry. These include S. Typhimurium, S. Enteritidis, S. Heidelberg, S. Montevideo, S. Saintpaul, and S. I 4,[5],12:i:-. PMID- 17911228 TI - Genotype by environment interaction for lamb weaning weight in two Norwegian sheep breeds. AB - Genotype x environment interaction (G x E) effects on live weaning weights of lambs were studied by using the 2 breeds Norwegian White sheep (NWS; heavy, long tailed) and Spel sheep (Spel; lighter, short-tailed) as genetic groups (G). A total of 37,338 NWS lambs and 30,075 Spel lambs born from 1989 to 1999 on 40 farms that kept both breeds together were included in the analyses. Environment was characterized by farm x year (E). In a mixed linear model framework, significance of the random G x E effect and breed-specific environmental variances were tested by using a log-likelihood approach. Directions and magnitudes of the effect were described through variance component estimates. An across-genotype environmental correlation was also used. There was a significant G x E effect on lamb BW; significant breed differences were found for variance of flock x year effects, indicating different phenotypic plasticities with changing flock x year environments, with the NWS being more sensitive to environmental change. Further, the breed-specific residual variance was greater for NWS, indicating that the effects of environmental variation were larger for the weaning weights of the NWS breed within flock and year. Further, the correlation between flock x year effects for the 2 breeds was significantly different from unity (0.82 +/- 0.02), indicating that the common environment is "perceived" differently in the 2 breeds. The best environment for one breed is not necessarily best for the other breed, and vice versa. Solutions of flock x year effects may be used to describe how environmental characteristics such as climate and topography affect the production of different genotypes, and for clustering of environments, thus facilitating improvement of breeding programs and management schemes for domestic and wild ungulate populations. PMID- 17911229 TI - Invited review: Role of livestock in human nutrition and health for poverty reduction in developing countries. AB - Livestock keeping is critical for many of the poor in the developing world, often contributing to multiple livelihood objectives and offering pathways out of poverty. Livestock keeping also affects an indispensable asset of the poor, their human capital, through its impact on their own nutrition and health. This paper outlines the linkages between livestock keeping and the physical well-being of the poor, and examines a number of commonly held beliefs that misrepresent livestock development issues related to these linkages. These beliefs limit the scope of intervention programs to promote livestock and limit their potential contribution to poverty reduction. Recognition of the complexity of the role livestock play in household decision-making and of the opportunities foregone due to these misconceptions can enhance the ability of livestock to contribute to human well-being in the developing world. PMID- 17911231 TI - Integrating the immune system with the regulation of growth and efficiency. AB - Muscle growth in meat animals is a complex process governed by integrated signals emanating from multiple endocrine and immune cells. A generalized phenomenon among meat animal industries is that animals commonly fail to meet their genetic potential for growth in commercial production settings. Recent evidence indicates that adipocytes and myofibers are equipped with functional pattern recognition receptors and are capable of responding directly to the corresponding pathogens and other receptor ligands. Thus, these cells are active participants in the innate immune response and, as such, produce a number of immune and metabolic regulators, including proinflammatory cytokines and adiponectin. Specifically, the transcription factor, nuclear factor kappa B, is activated in adipocytes and muscle cells by bacterial lipopolysaccharide and certain saturated fatty acids, which are potent agonists for the Toll-like receptor-4 pattern recognition receptor. Receptor activation results in the local production of interleukin-6 and tumor necrosis factor-alpha, and creates a local environment by which these cytokines regulate both metabolic and immunological pathways. However, adipocytes are also the predominant source of the antiinflammatory hormone, adiponectin, which suppresses the activation of nuclear factor kappa B and the production of proinflammatory cytokines. The molecular ability to recognize antigens and produce regulatory molecules strategically positions adipocytes and myofibers to regulate growth locally and to reciprocally regulate metabolism in peripheral tissues. PMID- 17911232 TI - Bioavailability of copper from copper glycinate in steers fed high dietary sulfur and molybdenum. AB - Sixty Angus (n = 29) and Angus-Sim-mental cross (n = 31) steers, averaging 9 mo of age and 277 kg of initial BW, were used in a 148-d study to determine the bioavailability of copper glycinate (CuGly) relative to feed-grade copper sulfate (CuSO(4)) when supplemented to diets high in S and Mo. Steers were blocked by weight within breed and randomly assigned to 1 of 5 treatments: 1) control (no supplemental Cu), 2) 5 mg of Cu/kg of DM from CuSO(4), 3) 10 mg of Cu/kg of DM from CuSO(4), 4) 5 mg of Cu/kg of DM from CuGly, and 5) 10 mg of Cu/kg of DM from CuGly. Steers were individually fed a corn silage-based diet (analyzed 8.2 mg of Cu/kg of DM), and supplemented with 2 mg of Mo/kg of diet DM and 0.15% S for 120 d (phase 1). Steers were then supplemented with 6 mg of Mo/kg of diet DM and 0.15% S for an additional 28 d (phase 2). Average daily gain and G:F were improved by Cu supplementation regardless of source (P = 0.01). Final ceruloplasmin, plasma Cu, and liver Cu values were greater (P < 0.05) in steers fed supplemental Cu compared with controls. Plasma Cu, liver Cu, and ceruloplasmin values were greater (P < 0.05) in steers supplemented with 10 mg of Cu/kg of DM vs. those supplemented with 5 mg of Cu/kg of DM. Based on multiple linear regression of final plasma Cu, liver Cu, and ceruloplasmin values on dietary Cu intake in phase 1 (2 mg of Mo/kg of DM), bioavailability of Cu from CuGly relative to CuSO(4) (100%) was 140 (P = 0.10), 131 (P = 0.12), and 140% (P = 0.01), respectively. Relative bio-availability of Cu from CuGly was greater than from CuSO(4) (P = 0.01; 144, 150, and 157%, based on plasma Cu, liver Cu, and ceruloplasmin, respectively) after supplementation of 6 mg of Mo/kg of DM for 28 d. Results of this study suggest that Cu from CuGly may be more available than CuSO(4) when supplemented to diets high in S and Mo. PMID- 17911234 TI - Gonadotropin-releasing hormone-induced ovulation and luteinizing hormone release in beef heifers: effect of day of the cycle. AB - The COSynch protocol has been used to synchronize ovulation and facilitate fixed time AI in beef cattle. Establishment and maintenance of pregnancy was negatively affected, in previous studies, by GnRH-induced ovulation of small dominant follicles (/=10 mm) and increased ovulatory response after GnRH 2. PMID- 17911235 TI - The interaction of harvesting time of day of switchgrass hay and ruminal degradability of supplemental protein offered to beef steers. AB - The objective of this study was to evaluate an interaction between harvest at 0600 (AM) vs. 1800 (PM) with high (HI) or low (LO) ruminal degradability of a protein supplement to change voluntary intake, digestion, or N retention by steers offered switchgrass (Panicum virgatum L.) hay. Black steers (255 +/- 14 kg of BW) were blocked by BW, and then randomly assigned (5 steers each) to AM/HI, PM/HI, AM/LO, or PM/LO treatment groups. Steers were group-housed in covered, outdoor pens with individual feeding gates. After adaptation and standardization, intake was measured for 21 d followed by a digestion trial (5 d of total collection). Steers were offered 767 (LO) or 825 (HI) g/d of supplement to provide 268 g of CP/d. Compared with AM, PM had greater (P = 0.01) concentrations of total nonstructural carbohydrate (TNC, 71 vs. 56 g/kg of DM), and lesser concentrations of NDF (760 vs. 770 g/kg of DM, P = 0.02), ADF (417 vs. 427 g/kg of DM, P = 0.02), and CP (55.9 vs. 58.6 g/ kg of DM, P = 0.07). Protein fractions A, B(2), and B(3) were similar for AM and PM, but HI contained more (P < 0.02) A (694 vs. 296 g/kg of protein) and less B(2) (174 vs. 554 g/kg of protein) fraction than LO. Harvest interacted with supplement to increase (P = 0.07) ad libitum digestible DMI for steers offered PM/HI (11.4 g/kg of BW daily) compared with steers offered PM/LO (10.2 g/kg of BW daily), but there was no difference for steers offered AM/LO or AM/HI (10.7 g/kg of BW). Apparent digestibilities of DM (594 vs. 571 g/kg of intake), NDF (591 vs. 562 g/kg of intake), ADF (585 vs. 566 g/kg of intake), and N (651 vs. 632 g/kg of intake) were greater (P < 0.04) for PM than for AM. Apparent digestibility of N was greater (P = 0.02) for HI (652 g/ kg of intake) vs. LO (631 g/kg of intake). Interactions between harvest and supplement for apparent digestibilities of NDF (P = 0.09) and ADF (P = 0.03) were due to no change or an increase in digestibility in response to increased ruminal degradability of supplement in steers offered PM harvest, whereas increased ruminal degradability of supplement decreased digestibility of NDF and ADF in steers offered AM harvest. Treatments did not affect hay intake (3.93 kg/d), N retained (15.8 g/d), or plasma urea N (5.25 mM) during ad libitum intake. Greater TNC in PM vs. AM harvest was not sufficient by itself to increase total voluntary DMI, but greater protein degradability interacted with harvest time to increase ruminal fiber digestibility and digestible DMI of beef steers offered PM vs. AM harvest. PMID- 17911236 TI - A new heat load index for feedlot cattle. AB - The ability to predict the effects of extreme climatic variables on livestock is important in terms of welfare and performance. An index combining temperature and humidity (THI) has been used for more than 4 decades to assess heat stress in cattle. However, the THI does not include important climatic variables such as solar load and wind speed (WS, m/s). Likewise, it does not include management factors (the effect of shade) or animal factors (genotype differences). Over 8 summers, a total of 11,669 Bos taurus steers, 2,344 B. taurus crossbred steers, 2,142 B. taurus x Bos indicus steers, and 1,595 B. indicus steers were used to develop and test a heat load index (HLI) for feedlot cattle. A new HLI incorporating black globe (BG) temperature ( degrees C), relative humidity (RH, decimal form), and WS was initially developed by using the panting score (PS) of 2,490 Angus steers. The HLI consists of 2 parts based on a BG temperature threshold of 25 degrees C: HLI(BG>25) = 8.62 + (0.38 x RH) + (1.55 x BG) - (0.5 x WS) + e((2.4-WS)), and HLI(BG<25) = 10.66 + (0.28 x RH) + (1.3 x BG) - WS, where e is the base of the natural logarithm. A threshold HLI above which cattle of different genotypes gain body heat was developed for 7 genotypes. The threshold for unshaded black B. taurus steers was 86, and for unshaded B. indicus (100%) the threshold was 96. Threshold adjustments were developed for factors such as coat color, health status, access to shade, drinking water temperature, and manure management. Upward and downward adjustments are possible; upward adjustments occur when cattle have access to shade (+3 to +7) and downward adjustments occur when cattle are showing clinical signs of disease (-5). A related measure, the accumulated heat load (AHL) model, also was developed after the development of the HLI. The AHL is a measure of the animal's heat load balance and is determined by the duration of exposure above the threshold HLI. The THI and THI-hours (hours above a THI threshold) were compared with the HLI and AHL. The relationships between tympanic temperature and the average HLI and THI for the previous 24 h were R(2) = 0.67, P < 0.001, and R(2) = 0.26, P < 0.001, respectively. The R(2) for the relationships between HLI or AHL and PS were positive (0.93 and 0.92 for HLI and AHL, respectively, P < 0.001). The R(2) for the relationship between THI and PS was 0.61 (P < 0.001), and for THI-hours was 0.37 (P < 0.001). The HLI and the AHL were successful in predicting PS responses of different cattle genotypes during periods of high heat load. PMID- 17911237 TI - Effects of grain processing and dietary lipid source on performance, carcass characteristics, plasma fatty acids, and sensory properties of steaks from finishing cattle. AB - An experiment was conducted to evaluate the effects of grain processing and lipid addition to finishing diets on cattle performance, carcass characteristics, and meat quality. Eighty Hereford x Angus steers (384 kg +/- 17 kg of BW) were fed diets containing steam-flaked corn (SFC) or dry-rolled corn (DRC) with and without the addition of tallow (SFC/Fat and DRC/Fat) or steam-flaked corn with ground flaxseed (SFC/Flax). Ribeye steaks from steers fed SFC, SFC/Fat, or SFC/ Flax were used to evaluate the effects of fat source on meat quality. Cattle fed SFC and SFC/Fat tended to have greater ADG, G:F, HCW, and USDA yield grade, compared with those fed DRC and DRC/Fat (P < 0.10). Steaks from steers fed SFC/Flax developed a detectable off-flavor (P < 0.05) compared with steaks from steers fed SFC and SFC/Fat, and steaks from steers fed SFC retained desirable color longer than those from steers fed SFC/Flax (P < 0.05). Feeding SFC/Flax increased deposition of alpha-linolenic acid in muscle tissue compared with feeding SFC or SFC/Fat (P < 0.01). Dietary treatment did not cause differences in tenderness, juiciness, or flavor intensity. Ground flaxseed can replace tallow in finishing diets without loss in performance, but flax may affect flavor and color stability of beef. Feeding flaxseed can effectively alter composition of carcass tissues to yield beef that is high in n-3 fatty acids. PMID- 17911239 TI - Genetic parameters for image analysis traits on M. longissimus thoracis and M. trapezius of carcass cross section in Japanese Black steers. AB - In Japan, the degree of marbling in ribeye (M. longissimus thoracis) is evaluated in the beef meat grading process. However, other muscles (e.g., M. trapezius) are also important in determining the meat quality and carcass market prices. The purpose of this study was to estimate genetic parameters for M. longissimus thoracis (M-LONG) and M. trapezius (M-TRAP) of carcass cross section of Japanese Black steers by computer image analysis. The number of records of Japanese Black steers and the number of pedigree records were 2,925 and 10,889, respectively. Digital images of the carcass cross section were taken between the sixth and seventh ribs by photographing equipment. Muscle area (MA), fat area ratio (FAR), overall coarseness of marbling particles (OCM), and coarseness of maximum marbling particle (MMC) in M-LONG and M-TRAP were calculated by image analysis. Genetic parameters for these traits were estimated using the AIREMLF90 program with an animal model. Fixed effects that were included in the model were dates of arrival at the carcass market and slaughter age (mo), and random effects of fattening farms, additive genetic effects and residuals were included in the model. For M-LONG, heritability estimates (+/-SE) were 0.46 +/- 0.06, 0.59 +/- 0.06, 0.47 +/- 0.06, and 0.20 +/- 0.05 for MA, FAR, OCM, and MMC, respectively. Heritability estimates (+/-SE) in M-TRAP were 0.47 +/- 0.06, 0.57 +/- 0.07, 0.49 +/- 0.07, and 0.13 +/- 0.04 for the same traits. Genetic correlations between subcutaneous fat thickness and FAR for M-LONG and M-TRAP were negative (-0.21 and -0.19, respectively). Those correlations between M-LONG and M-TRAP were moderate to high for MA, FAR, OCM, and MMC (0.38, 0.52, 0.39, and 0.60, respectively). These results indicate that other muscles including M-LONG should be evaluated for more efficient genetic improvement. PMID- 17911238 TI - Use of mannanoligosaccharides and zinc chelate as growth promoters and diarrhea preventative in weaning pigs: Effects on microbiota and gut function. AB - The efficacy of a commercial source of mannanoligosacharides (BM), organic zinc (BP), or their combination to enhance performance, gastrointestinal health, and immune response in weaned pigs was evaluated. A total of 128 piglets, weaned at 20 +/- 2 d, were housed in 32 pens. Animals received 1 of 4 dietary treatments: a control diet (CT) to which 0.2% of BM, 80 mg/kg of Zn as BP, or both additives (BMP) were added. The experiment lasted for 5 wk including a prestarter period of 2 wk and a starter period of 3 wk. Body weight was recorded and daily feed intake was calculated. Fecal consistency was monitored for the first 21 d. After 2 wk, 32 animals were killed, digesta samples from the stomach, ileum, and cecum were collected, and pH and the short-chain fatty acid profile were determined. Microbiological counts for enterobacteria and lactobacilli were evaluated using quantitative PCR. Histological parameters in the jejunum and immunoglobulin concentrations in serum and ileal digesta were also measured. Both additives improved G:F during the starter period (0.63, 0.69, 0.67, and 0.68 for CT, BM, BP, and BMP, respectively; P < 0.04). Mean fecal score values for the first 21 d were improved by BM and BP, showing decreased values compared with the CT diet (1.22, 0.89, 0.87, and 1.06 for CT, BM, BP, and BMP, respectively; P = 0.002). The addition of BM decreased enterobacteria counts in the jejunum (9.13, 8.05, 8.87, and 7.89 log 16S rRNA gene copies/g of matter for CT, BM, BP, and BMP, respectively; P = 0.05). Empty ileal weight, defined as the segment including the continuous Peyer's patch, tended (P = 0.08) to increase with BP treatment (8.9, 9.6, 11.9, and 10.3 g/kg of BW for CT, BM, BP, and BMP, respectively). Crypt depths in the jejunum were lower in animals fed the combination of the additives (BPM) compared with those fed the control diet (281 vs. 235; P < 0.03). No significant differences were registered in pH, short-chain fatty acids, or serum and ileal immunoglobulin concentrations. The results suggest that the use of BM or BP can improve the efficiency of gain during the starter period. PMID- 17911240 TI - Optimization of in vitro conditions for bovine subcutaneous and intramuscular preadipocyte differentiation. AB - The objective of these experiments was to develop an in vitro cell culture system for differentiation of bovine preadipocytes, which will permit examination of differences in differentiation between intramuscular (i.m.) and subcutaneous (s.c.) bovine preadipocytes. Stromal-vascular cells from bovine i.m. and s.c. adipose depots were isolated and cultured. Clonally derived s.c. preadipocytes were used to determine the ability of insulin, bovine serum lipids, octanoate, acetic acid, dexamethasone (DEX), and troglitazone (TRO) to elicit differentiation of these cells when added to serum-free medium. Addition of 10 and 20 microL/mL of a commercially available serum lipids supplement to low glucose Dulbecco's modified Eagle's medium containing 280 nM insulin increased glycerol-3-phosphate dehydrogenase (GPDH) activity (P < 0.01). Inclusion of 1.25 to 10 microM TRO to medium containing 280 nM insulin and 20 microL/ mL serum lipids supplement also increased GPDH activity (P < 0.001) compared with 0 microM TRO. The combination of 280 nM insulin, 1 mM octanoate, and 10 mM acetic acid, with 48 h exposure to 0.25 microM DEX caused morphological differentiation in a small number of cells but did not stimulate GPDH activity (P = 0.99). When used together, 280 nM insulin, 20 microL/mL of serum lipids supplement, 40 microM TRO, and 0.25 microM DEX stimulated differentiation compared with the aforementioned treatment (P < 0.001). Omission of TRO or insulin from this medium reduced GPDH activity by 68% (P < 0.001), whereas removal of DEX tended to reduce GPDH activity (P = 0.06). Preadipocytes from s.c. (n = 3) and i.m. (n = 2) adipose tissues of 3 steers were used to determine the effects of TRO on differentiation using the established conditions. Forty to sixty microM TRO enhanced differentiation compared with 0 microM TRO (P < 0.02) in both depots. No depot differences in response to TRO were detected (P = 0.32). These data demonstrate that bovine preadipocytes are capable of differentiation in response to combinations of insulin, serum lipids, DEX, and TRO. Although TRO enhanced differentiation of bovine preadipocytes, no differential effects of TRO on the differentiation of s.c. and i.m. cells were detected. PMID- 17911241 TI - Effect of live yeast culture supplementation on apparent digestibility and rate of passage in horses fed a high-fiber or high-starch diet. AB - Eight crossbred male horses aged 12 +/- 5 yr and with BW of 305 +/- 18 kg were used in pairs in a 4 x 4 Latin square design with 4 ground and pelleted diets. Each pair included a cecum and right ventral colon-fistulated animal and a cecal fistulated animal. The 4 horse diets were a high-fiber diet (HF+0) based on dehydrated alfalfa, a high-starch diet based on barley and wheat bran (HS+0), and the HF or HS diets supplemented with Saccharomyces cerevisiae (SC) CBS 493.94 (HF+SC and HS+SC). The probiotic preparation contained 4.5 x 10(9) cfu/g of live yeast mixed with the culture medium, and was top-dressed onto the feed pellets at a rate of 10 g/d, equally distributed between the 2 daily meals. All 4 diets were offered in the same quantities (18.0 g of pelleted feed DM + 3.5 g of long wheat straw/kg of BW per d). Each of the 4 experimental treatments was divided into a 21-d period of diet adaptation followed by a 10-d period of total fecal collection for digesta flow rate and apparent digestibility measurements. Three markers were used to measure mean retention time (MRT) of the feed particles: Yb bound to the pelleted feeds for MRT in the whole digestive tract (MRT(Yb)), Eu bound to the pelleted feeds, and Dy bound to the fecal particles for MRT in the hindgut (MRT(Eu) and MRT(Dy)). Apparent digestibilities of DM, OM, and CP were greater (P < 0.001) in the HS than HF diet, independently of SC supplementation, whereas ADF digestibility was greatest in the HF diet (P = 0.035). Cellulolytic activity estimated through the in vitro disappearance rate of the dietary ADF fraction (IVAD(ADF)) was less (P < 0.001) in the HS than the HF diet. There was no dietary effect on NDF digestibility due to the longer MRT(Eu) of small particles in the hindgut (P = 0.036), which compensated for the lower fibrolytic activity expressed per unit of time in the HS compared with the HF diet. Supplementation with SC improved ADF digestibility (P = 0.038) and stimulated DM (P = 0.030) and NDF (P = 0.038) intakes, but had no effect on the MRT of solid digesta. The absence of any significant diet x SC interaction supports the strategy of using SC to stimulate cellulose digestion and improve the nutritional status of horses under both HF and HS diets. PMID- 17911242 TI - A surface on the androgen receptor that allosterically regulates coactivator binding. AB - Current approaches to inhibit nuclear receptor (NR) activity target the hormone binding pocket but face limitations. We have proposed that inhibitors, which bind to nuclear receptor surfaces that mediate assembly of the receptor's binding partners, might overcome some of these limitations. The androgen receptor (AR) plays a central role in prostate cancer, but conventional inhibitors lose effectiveness as cancer treatments because anti-androgen resistance usually develops. We conducted functional and x-ray screens to identify compounds that bind the AR surface and block binding of coactivators for AR activation function 2 (AF-2). Four compounds that block coactivator binding in solution with IC(50) approximately 50 microM and inhibit AF-2 activity in cells were detected: three nonsteroidal antiinflammatory drugs and the thyroid hormone 3,3',5 triiodothyroacetic acid. Although visualization of compounds at the AR surface reveals weak binding at AF-2, the most potent inhibitors bind preferentially to a previously unknown regulatory surface cleft termed binding function (BF)-3, which is a known target for mutations in prostate cancer and androgen insensitivity syndrome. X-ray structural analysis reveals that 3,3',5-triiodothyroacetic acid binding to BF-3 remodels the adjacent interaction site AF-2 to weaken coactivator binding. Mutation of residues that form BF-3 inhibits AR function and AR AF-2 activity. We propose that BF-3 is a previously unrecognized allosteric regulatory site needed for AR activity in vivo and a possible pharmaceutical target. PMID- 17911244 TI - An interpretation of fluctuations in enzyme catalysis rate, spectral diffusion, and radiative component of lifetimes in terms of electric field fluctuations. AB - Time-dependent fluctuations in the catalysis rate (deltak(t)) observed in single enzyme experiments were found in a particular study to have an autocorrelation function decaying on the same time scale as that of spectral diffusion deltaomega(0)(t). To interpret this similarity, the present analysis focuses on a factor in enzyme catalysis, the local electrostatic interaction energy (E) at the active site and its effect on the activation free energy barrier. We consider the slow fluctuations of the electrostatic interaction energy (deltaE(t)) as a contributor to deltak(t) and relate the latter to deltaomega(0)(t). The resulting relation between deltak(t) and deltaomega(0)(t) is a dynamic analog of the solvatochromism used in interpreting solvent effects on organic reaction rates. The effect of the postulated deltaE(t) on fluctuations in the radiative component (deltagamma(r)(-1)(t)) of the fluorescence decay of chromophores in proteins also is examined, and a relation between deltagamma(r)(-1)(t) and deltaomega(0)(t) is obtained. Experimental tests will determine whether the correlation functions for deltak(t), deltaomega(0)(t), and deltagamma(r)(-1) are indeed similar for any enzyme. Measurements of dielectric dispersion, epsilon(omega), for the enzyme discussed elsewhere will provide further insight into the correlation function for deltaE(t). They also will determine whether fluctuations in the nonradiative component gamma(nr)(-1) of the lifetime decay has a different origin, fluctuations in distance for example. PMID- 17911243 TI - Deletion mutations in N-terminal alpha1 helix render heat labile enterotoxin B subunit susceptible to degradation. AB - Heat-labile enterotoxin (LT) from enterotoxigenic Escherichia coli is a heterohexameric protein consisting of an enzymatically active A subunit, LTA, and a carrier pentameric B subunit, LTB. It is clear from the crystal structure of LTB that the N-terminal alpha1 helix lies outside the core structure. However, the function of the N-terminal alpha1 helix of LTB is unknown. The present work was carried out to investigate the effect of site-directed mutagenesis of the alpha1 helix on LTB synthesis. Six amino acids (PQSITE) located at positions 2-7 from the N terminus, including 4 aa from the alpha1 helix, were deleted by site directed mutagenesis. The deletion resulted in complete inhibition of LTB expression in E. coli when expressed along with its signal sequence. A single amino acid deletion within the alpha1 helix also resulted in loss of expression. However, a single amino acid deletion outside the alpha1 helix did not affect LTB synthesis. Mutant proteins, whose synthesis was not detected in vivo, could be successfully translated in vitro by using the coupled transcription-translation system. Immunoblot analysis, Northern blot analysis, and in vitro transcription translation data collectively indicate that the lack of synthesis of the mutant proteins is caused by the immediate degradation of the expressed product by cellular proteases rather than by faulty translation of mutant LTB mRNA. Coexpression of the LTA could not rescue the degradation of LTB mutants. PMID- 17911245 TI - Plasma sodium stiffens vascular endothelium and reduces nitric oxide release. AB - Dietary salt plays a major role in the regulation of blood pressure, and the mineralocorticoid hormone aldosterone controls salt homeostasis and extracellular volume. Recent observations suggest that a small increase in plasma sodium concentration may contribute to the pressor response of dietary salt. Because endothelial cells are (i) sensitive to aldosterone, (ii) in physical contact with plasma sodium, and (iii) crucial regulators of vascular tone, we tested whether acute changes in plasma sodium concentration, within the physiological range, can alter the physical properties of endothelial cells. The tip of an atomic force microscope was used as a nanosensor to measure stiffness of living endothelial cells incubated for 3 days in a culture medium containing aldosterone at a physiological concentration (0.45 nM). Endothelial cell stiffness was unaffected by acute changes in sodium concentration <135 mM but rose steeply between 135 and 145 mM. The increase in stiffness occurred within minutes. Lack of aldosterone in the culture medium or treatment with the epithelial sodium channel inhibitor amiloride prevented this response. Nitric oxide formation was found down regulated in cells cultured in aldosterone-containing high sodium medium. The results suggest that changes in plasma sodium concentration per se may affect endothelial function and thus control vascular tone. PMID- 17911246 TI - Adaptations of guest and host in expanded self-assembled capsules. AB - Reversible encapsulation complexes create spaces where two or more molecules can be temporarily isolated. When the mobility of encapsulated molecules is restricted, different arrangements in space are possible, and new forms of isomerism ("social isomerism") are created: the orientation of one encapsulated molecule influences that of the other in the confined space. Expansion of a capsule's length is possible through addition of small-molecule spacer elements. The expanded capsules have dimensions that permit the observation of social isomerism of two identical guests, and they adopt arrangements that properly fill the host's space. The host also can adapt to longer guests by incorporating additional spacers, much as protein modules are added to a viral capsid in response to larger genomes. Arachidonic and related fatty acid derivatives act in this way to induce the assembly of further extended capsules having sufficient length to accommodate them. PMID- 17911247 TI - Ca2+ influx is an essential component of the positive-feedback loop that maintains leading-edge structure and activity in macrophages. AB - In migrating eukaryotic cells, phosphatidylinositol 3-kinase (PI3K), filamentous actin (F-actin), and monomeric Rho GTPases are key components of a complex positive-feedback system that maintains and amplifies a phosphatidylinositol 3,4,5-trisphosphate signal at the leading edge of the cell. This lipid signal is required for cell polarization and movement. In leukocytes and Dictyostelium, activation or inhibition of any one of these components leads to the activation or inhibition, respectively, of the others via undefined feedback interactions. The role of Ca(2+) signals in migrating leukocytes is controversial, and there has been no indication that Ca(2+) participates in positive feedback. Here, we demonstrate that an extracellular Ca(2+) influx is required for positive feedback at the leading edge of spontaneously polarized macrophages. Inhibition of extracellular Ca(2+) influx leads to loss of leading-edge PI3K activity, disassembly of F-actin, cessation of ruffling, and decay of chemoattractant signals. Conversely, increasing cytosolic Ca(2+) enhances membrane ruffling, PI3K activity, and F-actin accumulation. Overall, these findings demonstrate that an extracellular Ca(2+) influx is an essential component, together with PI3K and F actin, of the positive-feedback cycle that maintains leading-edge structure and ruffling activity and that supports the chemoattractant response. Strikingly, the Ca(2+)-sensitive enzyme protein kinase Calpha (PKCalpha) is enriched at the leading edge, and its enrichment is sensitive to blockade of Ca(2+) influx, to inhibition of PI3K activity, and to F-actin depolymerization. These findings support the working hypothesis that a local, leading-edge Ca(2+) signal recruits PKCalpha as a central player in the positive-feedback loop. PMID- 17911248 TI - A mechanobiochemical mechanism for monooriented chromosome oscillation in mitosis. AB - During mitosis, the condensed chromosomes undergo a series of spectacular oscillations after they are captured in an end-on manner by kinetochore microtubules (KMT) emanating from the spindle poles. Such oscillations are commonly attributed to tug-of-war-like mechanisms, where the mechanical force imbalance alone drives the chromosome movement. However, a large portion of the force imbalance upon the chromosome is absorbed by the kinetochore and may not drive chromosome movement directly. Mounting evidence suggests that such resistance by the kinetochores regulates the chemical reactions of KMT plus-end growth and shrinkage, which have been shown as the determinant of the chromosome antipoleward (AP) and poleward movements. Here we incorporate this important regulatory feature, propose a mechanobiochemical feedback mechanism, and apply it to the monooriented chromosome oscillation, the early stage of the series of observed chromosome oscillations. In this model, the mechanical movement of the chromosome and the local biochemical reactions at the attached kinetochore region form a feedback loop that drives the oscillation. The force imbalance exerted on the chromosomes provides a bias (via mechanically sensitive proteins) on the local biochemical reactions controlling the KMT plus-end dynamics, and the movement of the chromosome in turn changes the forces exerted on it through the experimentally supported gradient in AP force. The proposed feedback mechanism can generate oscillatory behavior that depends on the topology of the feedback loop but is largely independent of the detailed molecular mechanism. We suggest potential molecular players, whose perturbation may allow direct experimental tests of the model. PMID- 17911249 TI - Skewed association of polyfunctional antigen-specific CD8 T cell populations with HLA-B genotype. AB - We studied CD8 T cell responses against HIV-1, cytomegalovirus, Epstein-Barr virus, and influenza in 128 subjects and demonstrate that polyfunctional CD8 T cell responses, also including IL-2 production and Ag-specific proliferation, are predominantly driven by virus epitopes restricted by HLA-B alleles. Interestingly, these protective CD8 T cells are equipped with low-avidity T cell receptors (TCRs) for the cognate virus epitope. Conversely, HLA-A-restricted epitopes are mostly associated with "only effector" IFN-gamma-secreting, with cytotoxicity, and with the lack of IL-2 production and Ag-specific proliferation. These CD8 T cells are equipped with high-avidity TCR and express higher levels of the T cell exhaustion marker PD-1. Thus, the functional profile of the CD8 T cell response is strongly influenced by the extent to which there is stimulation of polyfunctional (predominantly restricted by HLA-B) versus only effector (restricted by HLA-A) T cell responses. These results provide the rationale for the observed protective role of HLA-B in HIV-1-infection and new insights into the relationship between TCR avidity, PD-1 expression, and the functional profile of CD8 T cells. PMID- 17911250 TI - Crystal structure of the Helicobacter pylori vacuolating toxin p55 domain. AB - Helicobacter pylori VacA, a pore-forming toxin secreted by an autotransporter pathway, causes multiple alterations in human cells, contributes to the pathogenesis of peptic ulcer disease and gastric cancer, and is a candidate antigen for inclusion in an H. pylori vaccine. Here, we present a 2.4-A crystal structure of the VacA p55 domain, which has an important role in mediating VacA binding to host cells. The structure is predominantly a right-handed parallel beta-helix, a feature that is characteristic of autotransporter passenger domains but unique among known bacterial protein toxins. Notable features of VacA p55 include disruptions in beta-sheet contacts that result in five beta-helix subdomains and a C-terminal domain that contains a disulfide bond. Analysis of VacA protein sequences from unrelated H. pylori strains, including m1 and m2 forms of VacA, allows us to identify structural features of the VacA surface that may be important for interactions with host receptors. Docking of the p55 structure into a 19-A cryo-EM map of a VacA dodecamer allows us to propose a model for how VacA monomers assemble into oligomeric structures capable of membrane channel formation. PMID- 17911251 TI - Gambogic amide, a selective agonist for TrkA receptor that possesses robust neurotrophic activity, prevents neuronal cell death. AB - Nerve growth factor (NGF) binds to TrkA receptor and triggers activation of numerous signaling cascades, which play critical roles in neuronal plasticity, survival, and neurite outgrowth. To mimic NGF functions pharmacologically, we developed a high-throughput screening assay to identify small-molecule agonists for TrkA receptor. The most potent compound, gambogic amide, selectively binds to TrkA, but not TrkB or TrkC, and robustly induces its tyrosine phosphorylation and downstream signaling activation, including Akt and MAPKs. Further, it strongly prevents glutamate-induced neuronal cell death and provokes prominent neurite outgrowth in PC12 cells. Gambogic amide specifically interacts with the cytoplasmic juxtamembrane domain of TrkA receptor and triggers its dimerization. Administration of this molecule in mice substantially diminishes kainic acid triggered neuronal cell death and decreases infarct volume in the transient middle cerebral artery occlusion model of stroke. Thus, gambogic amide might not only establish a powerful platform for dissection of the physiological roles of NGF and TrkA receptor but also provide effective treatments for neurodegenerative diseases and stroke. PMID- 17911252 TI - Alpha2-chimaerin interacts with EphA4 and regulates EphA4-dependent growth cone collapse. AB - EphA4-dependent growth cone collapse requires reorganization of actin cytoskeleton through coordinated activation of Rho family GTPases. Whereas various guanine exchange factors have recently been identified to be involved in EphA4-mediated regulation of Rho GTPases and growth cone collapse, the functional roles of GTPase-activating proteins in the process are largely unknown. Here we report that EphA4 interacts with alpha2-chimaerin through its Src homology 2 domain. Activated EphA4 induces a rapid increase of tyrosine phosphorylation of alpha2-chimaerin and enhances its GTPase-activating protein activity toward Rac1. More importantly, alpha2-chimaerin regulates the action of EphA4 in growth cone collapse through modulation of Rac1 activity. Our findings have therefore identified a new alpha2-chimaerin-dependent signaling mechanism through which EphA4 transduces its signals to the actin cytoskeleton and modulates growth cone morphology. PMID- 17911253 TI - Supermodeled sabercat, predatory behavior in Smilodon fatalis revealed by high resolution 3D computer simulation. AB - The American sabercat Smilodon fatalis is among the most charismatic of fossil carnivores. Despite broad agreement that its extraordinary anatomy reflects unique hunting techniques, after >150 years of study, many questions remain concerning its predatory behavior. Were the "sabers" used to take down large prey? Were prey killed with an eviscerating bite to the abdomen? Was its bite powerful or weak compared with that of modern big cats? Here we quantitatively assess the sabercat's biomechanical performance using the most detailed computer reconstructions yet developed for the vertebrate skull. Our results demonstrate that bite force driven by jaw muscles was relatively weak in S. fatalis, one third that of a lion (Panthera leo) of comparable size, and its skull was poorly optimized to resist the extrinsic loadings generated by struggling prey. Its skull is better optimized for bites on restrained prey where the bite is augmented by force from the cervical musculature. We conclude that prey were brought to ground and restrained before a killing bite, driven in large part by powerful cervical musculature. Because large prey is easier to restrain if its head is secured, the killing bite was most likely directed to the neck. We suggest that the more powerful jaw muscles of P. leo may be required for extended, asphyxiating bites and that the relatively low bite forces in S. fatalis might reflect its ability to kill large prey more quickly, avoiding the need for prolonged bites. PMID- 17911254 TI - Gel-forming mucins appeared early in metazoan evolution. AB - Mucins are proteins that cover and protect epithelial cells and are characterized by domains rich in proline, threonine, and serine that are heavily glycosylated (PTS or mucin domains). Because of their sequence polymorphism, these domains cannot be used for evolutionary analysis. Instead, we have made use of the von Willebrand D (VWD) and SEA domains, typical for mucins. A number of animal genomes were examined for these domains to identify mucin homologues, and domains of the resulting proteins were used in phylogenetic studies. The frog Xenopus tropicalis stands out because the number of gel-forming mucins has markedly increased to at least 25 as compared with 5 for higher animals. Furthermore, the frog Muc2 homologues contain unique PTS domains where cysteines are abundant. This animal also has a unique family of secreted mucin-like proteins with alternating PTS and SEA domains, a type of protein also identified in the fishes. The evolution of the Muc4 mucin seems to have occurred by recruitment of a PTS domain to AMOP, NIDO, and VWD domains from a sushi domain-containing family of proteins present in lower animals, and Xenopus is the most deeply branching animal where a protein similar to the mammalian Muc4 was identified. All transmembrane mucins seem to have appeared in the vertebrate lineage, and the MUC1 mucin is restricted to mammals. In contrast, proteins with properties of the gel-forming mucins were identified also in the starlet sea anemone Nematostella vectensis, demonstrating an early origin of this group of mucins. PMID- 17911255 TI - Activin A is a critical component of the inflammatory response, and its binding protein, follistatin, reduces mortality in endotoxemia. AB - Activin A is a member of the transforming growth factor-beta superfamily, which we have identified as having a role in inflammatory responses. We show that circulating levels of activin increase rapidly after LPS-induced challenge through activation of Toll-like receptor 4 and the key adaptor protein, MyD88. Treatment with the activin-binding protein, follistatin, alters the profiles of TNF, IL-1beta, and IL-6 after LPS stimulation, indicating that activin modulates the release of several key proinflammatory cytokines. Further, mice administered one 10-mug dose of follistatin to block activin effects have increased survival after a lethal dose of LPS, and the circulating levels of activin correlate with survival outcome. These findings demonstrate activin A's crucial role in the inflammatory response and show that blocking its actions by the use of follistatin has significant therapeutic potential to reduce the severity of inflammatory diseases. PMID- 17911256 TI - Control of electrostatic interactions between F-actin and genetically modified lysozyme in aqueous media. AB - The aim for deterministic control of the interactions between macroions in aqueous media has motivated widespread experimental and theoretical work. Although it has been well established that like-charged macromolecules can aggregate under the influence of oppositely charged condensing agents, the specific conditions for the stability of such aggregates can only be determined empirically. We examine these conditions, which involve an interplay of electrostatic and osmotic effects, by using a well defined model system composed of F-actin, an anionic rod-like polyelectrolyte, and lysozyme, a cationic globular protein with a charge that can be genetically modified. The structure and stability of actin-lysozyme complexes for different lysozyme charge mutants and salt concentrations are examined by using synchrotron x-ray scattering and molecular dynamics simulations. We provide evidence that supports a structural transition from columnar arrangements of F-actin held together by arrays of lysozyme at the threefold interstitial sites of the actin sublattice to marginally stable complexes in which lysozyme resides at twofold bridging sites between actin. The reduced stability arises from strongly reduced partitioning of salt between the complex and the surrounding solution. Changes in the stability of actin-lysozyme complexes are of biomedical interest because their formation has been reported to contribute to the persistence of airway infections in cystic fibrosis by sequestering antimicrobials such as lysozyme. We present x-ray microscopy results that argue for the existence of actin-lysozyme complexes in cystic fibrosis sputum and demonstrate that, for a wide range of salt conditions, charge-reduced lysozyme is not sequestered in ordered complexes while retaining its bacterial killing activity. PMID- 17911257 TI - Guanine riboswitch variants from Mesoplasma florum selectively recognize 2' deoxyguanosine. AB - Several mRNA aptamers have been identified in Mesoplasma florum that have sequence and structural features resembling those of guanine and adenine riboswitches. Two features distinguish these RNAs from established purine-sensing riboswitches. All possess shortened hairpin-loop sequences expected to alter tertiary contacts known to be critical for aptamer folding. The RNAs also carry nucleotide changes in the core of each aptamer that otherwise is strictly conserved in guanine and adenine riboswitches. Some aptamers retain the ability to selectively bind guanine or adenine despite these mutations. However, one variant type exhibits selective and high-affinity binding of 2'-deoxyguanosine, which is consistent with its occurrence in the 5' untranslated region of an operon containing ribonucleotide reductase genes. The identification of riboswitch variants that bind nucleosides and reject nucleobases reveals that natural metabolite-sensing RNA motifs can accrue mutations that expand the diversity of ligand detection in bacteria. PMID- 17911258 TI - Toxofilin from Toxoplasma gondii forms a ternary complex with an antiparallel actin dimer. AB - Many human pathogens exploit the actin cytoskeleton during infection, including Toxoplasma gondii, an apicomplexan parasite related to Plasmodium, the agent of malaria. One of the most abundantly expressed proteins of T. gondii is toxofilin, a monomeric actin-binding protein (ABP) involved in invasion. Toxofilin is found in rhoptry and presents an N-terminal signal sequence, consistent with its being secreted during invasion. We report the structure of toxofilin amino acids 69-196 in complex with the host mammalian actin. Toxofilin presents an extended conformation and interacts with an antiparallel actin dimer, in which one of the actins is related by crystal symmetry. Consistent with this observation, analytical ultracentrifugation analysis shows that toxofilin binds two actins in solution. Toxofilin folds into five consecutive helices, which form three relatively independent actin-binding sites. Helices 1 and 2 bind the symmetry related actin molecule and cover its nucleotide-binding cleft. Helices 3-5 bind the other actin and constitute the primary actin-binding region. Helix 3 interacts in the cleft between subdomains 1 and 3, a common binding site for most ABPs. Helices 4 and 5 wrap around actin subdomain 4, and residue Gln-134 of helix 4 makes a hydrogen-bonding contact with the nucleotide in actin, both of which are unique features among ABPs. Toxofilin dramatically inhibits nucleotide exchange on two actin molecules simultaneously. This effect is linked to the formation of the antiparallel actin dimer because a construct lacking helices 1 and 2 binds only one actin and inhibits nucleotide exchange less potently. PMID- 17911259 TI - Time course and calcium dependence of transmitter release at a single ribbon synapse. AB - At the first synapse in the auditory pathway, the receptor potential of mechanosensory hair cells is converted into a firing pattern in auditory nerve fibers. For the accurate coding of timing and intensity of sound signals, transmitter release at this synapse must occur with the highest precision. To measure directly the transfer characteristics of the hair cell afferent synapse, we implemented simultaneous whole-cell recordings from mammalian inner hair cells (IHCs) and auditory nerve fiber terminals that typically receive input from a single ribbon synapse. During a 1-s IHC depolarization, the synaptic response depressed >90%, representing the main source for adaptation in the auditory nerve. Synaptic depression was slightly affected by alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid receptor desensitization; however, it was mostly caused by reduced vesicular release. When the transfer function between transmitter release and Ca(2+) influx was tested at constant open probability for Ca(2+) channels (potentials >0 mV), a super linear relation was found. This relation is presumed to result from the cooperative binding of three to four Ca(2+) ions at the Ca(2+) sensor. However, in the physiological range for receptor potentials (-50 to -30 mV), the relation between Ca(2+) influx and afferent activity was linear, assuring minimal distortion in the coding of sound intensity. Changes in Ca(2+) influx caused an increase in release probability, but not in the average size of multivesicular synaptic events. By varying Ca(2+) buffering in the IHC, we further investigate how Ca(2+) channel and Ca(2+) sensor at this synapse might relate. PMID- 17911260 TI - Human combinatorial libraries yield rare antibodies that broadly neutralize hepatitis C virus. AB - One way to dissect the antibody response to an invading microorganism is to clone the antibody repertoire from immune donors and subsequently characterize the specific antibodies. Recently, methodological advances have allowed investigations of neutralizing antibodies against hepatitis C virus (HCV) in vitro. We have investigated three human mAbs, previously isolated from an individual infected with HCV of genotype 2b, that are known to cross-react in a binding assay to the envelope E2 protein of genotypes 1a and 1b. We now report that two of them have a neutralizing activity with a breadth not previously observed. Indeed, mAbs 1:7 and A8 recognized E2 from all of the six major genotypes, and they neutralized retroviral pseudoparticles [HCV pseudoparticles (HCVpp)] carrying genetically equally diverse HCV envelope glycoproteins. Importantly, these antibodies were also able to neutralize the cell culture infectious HCV clone JFH-1 in vitro, with IC(50) values of 60 ng/ml and 560 ng/ml, respectively. The conformational epitopes of these two broadly reactive antibodies were overlapping yet distinct and involved amino acid residues in the 523-535 region of E2, known to be important for the E2-CD81 interaction. The third antibody clone, representing a dominant population in the initial screen for these antibodies, was less broadly reactive and was unable to neutralize the genotype 2a infectious clone JFH-1. Our results confirm at the clonal level that broadly neutralizing human anti-HCV antibodies can be elicited and that the region amino acids 523-535 of the HCV envelope glycoprotein E2 carries neutralizing epitopes conserved across all genotypes. PMID- 17911262 TI - Native language governs interpretation of salient speech sound differences at 18 months. AB - One of the first steps infants take in learning their native language is to discover its set of speech-sound categories. This early development is shown when infants begin to lose the ability to differentiate some of the speech sounds their language does not use, while retaining or improving discrimination of language-relevant sounds. However, this aspect of early phonological tuning is not sufficient for language learning. Children must also discover which of the phonetic cues that are used in their language serve to signal lexical distinctions. Phonetic variation that is readily discriminable to all children may indicate two different words in one language but only one word in another. Here, we provide evidence that the language background of 1.5-year-olds affects their interpretation of phonetic variation in word learning, and we show that young children interpret salient phonetic variation in language-specific ways. Three experiments with a total of 104 children compared Dutch- and English learning 18-month-olds' responses to novel words varying in vowel duration or vowel quality. Dutch learners interpreted vowel duration as lexically contrastive, but English learners did not, in keeping with properties of Dutch and English. Both groups performed equivalently when differentiating words varying in vowel quality. Thus, at one and a half years, children's phonological knowledge already guides their interpretation of salient phonetic variation. We argue that early phonological learning is not just a matter of maintaining the ability to distinguish language-relevant phonetic cues. Learning also requires phonological interpretation at appropriate levels of linguistic analysis. PMID- 17911261 TI - Structure of outer membrane protein G by solution NMR spectroscopy. AB - The bacterial outer membrane protein G (OmpG), a monomeric pH-gated porin, was overexpressed in Escherichia coli and refolded in beta-octyl glucoside micelles. After transfer into dodecylphosphocholine micelles, the solution structure of OmpG was determined by solution NMR spectroscopy at pH 6.3. Complete backbone assignments were obtained for 234 of 280 residues based on CA, CB, and CO connection pathways determined from a series of TROSY-based 3D experiments at 800 MHz. The global fold of the 14-stranded beta-barrel was determined based on 133 long-range NOEs observed between neighboring strands and local chemical shift and NOE information. The structure of the barrel is very similar to previous crystal structures, but the loops of the solution structure are quite flexible. PMID- 17911263 TI - Transmission of a fatal clonal tumor by biting occurs due to depleted MHC diversity in a threatened carnivorous marsupial. AB - A fatal transmissible tumor spread between individuals by biting has emerged in the Tasmanian devil (Sarcophilus harrisii), a carnivorous marsupial. Here we provide genetic evidence establishing that the tumor is clonal and therefore foreign to host devils. Thus, the disease is highly unusual because it is not just a tumor but also a tissue graft, passed between individuals without invoking an immune response. The MHC plays a key role in immune responses to both tumors and grafts. The most common mechanism of immune evasion by tumors is down regulation of classical cell surface MHC molecules. Here we show that this mode of immune escape does not occur. However, because the tumor is a graft, it should still be recognized and rejected by the host's immune system due to foreign cell surface antigens. Mixed lymphocyte responses showed a lack of alloreactivity between lymphocytes of different individuals in the affected population, indicating a paucity of MHC diversity. This result was verified by genotyping, providing a conclusive link between a loss of MHC diversity and spread of a disease through a wild population. This novel disease arose as a direct result of loss of genetic diversity and the aggressive behavior of the host species. The neoplastic clone continues to spread although the population, and, without active disease control by removal of affected animals and the isolation of disease-free animals, the Tasmanian devil faces extinction. PMID- 17911265 TI - Force fluctuations and polymerization dynamics of intracellular microtubules. AB - Microtubules are highly dynamic biopolymer filaments involved in a wide variety of biological processes including cell division, migration, and intracellular transport. Microtubules are very rigid and form a stiff structural scaffold that resists deformation. However, despite their rigidity, inside of cells they typically exhibit significant bends on all length scales. Here, we investigate the origin of these bends using a Fourier analysis approach to quantify their length and time dependence. We show that, in cultured animal cells, bending is suppressed by the surrounding elastic cytoskeleton, and even large intracellular forces only cause significant bending fluctuations on short length scales. However, these lateral bending fluctuations also naturally cause fluctuations in the orientation of the microtubule tip. During growth, these tip fluctuations lead to microtubule bends that are frozen-in by the surrounding elastic network. This results in a persistent random walk of the microtubule, with a small apparent persistence length of approximately 30 microm, approximately 100 times smaller than that resulting from thermal fluctuations alone. Thus, large nonthermal forces govern the growth of microtubules and can explain the highly curved shapes observed in the microtubule cytoskeleton of living cells. PMID- 17911264 TI - Tumor protein 53-induced nuclear protein 1 expression is repressed by miR-155, and its restoration inhibits pancreatic tumor development. AB - Pancreatic cancer is a disease with an extremely poor prognosis. Tumor protein 53 induced nuclear protein 1 (TP53INP1) is a proapoptotic stress-induced p53 target gene. In this article, we show by immunohistochemical analysis that TP53INP1 expression is dramatically reduced in pancreatic ductal adenocarcinoma (PDAC) and this decrease occurs early during pancreatic cancer development. TP53INP1 reexpression in the pancreatic cancer-derived cell line MiaPaCa2 strongly reduced its capacity to form s.c., i.p., and intrapancreatic tumors in nude mice. This anti-tumoral capacity is, at least in part, due to the induction of caspase 3 mediated apoptosis. In addition, TP53INP1(-/-) mouse embryonic fibroblasts (MEFs) transformed with a retrovirus expressing E1A/ras(V12) oncoproteins developed bigger tumors than TP53INP1(+/+) transformed MEFs or TP53INP1(-/-) transformed MEFs with restored TP53INP1 expression. Finally, TP53INP1 expression is repressed by the oncogenic micro RNA miR-155, which is overexpressed in PDAC cells. TP53INP1 is a previously unknown miR-155 target presenting anti-tumoral activity. PMID- 17911266 TI - Modulation of LMP1 protein expression by EBV-encoded microRNAs. AB - Epstein-Barr virus (EBV) was the first human virus found to encode microRNAs (miRNAs), but the function of these miRNAs has been obscure. Nasopharyngeal carcinoma (NPC) is associated with EBV infection, and the EBV-encoded LMP1 is believed to be a key factor in NPC development. However, detection of LMP1 protein in NPC is variable. Here, we report that EBV-encoded BART miRNAs target the 3' UTR of the LMP1 gene and negatively regulate LMP1 protein expression. These miRNAs also modulate LMP1-induced NF-kappaB signaling and alleviate the cisplatin sensitivity of LMP1-expressing NPC cells. Consistent with a previous study on the NPC C666-1 cell line and C15 xenograft, we found abundant expression of BART miRNAs in NPC tissues. Furthermore, DNA sequencing revealed that the 3' UTR of LMP1 is highly conserved in NPC-derived EBV isolates. The data provide insight into the discrepancy between LMP1 transcript and protein detection in NPC and highlight the role of the EBV miRNAs in regulating LMP1 downstream signaling to promote cancer development. PMID- 17911267 TI - Activation of the PTEN/mTOR/STAT3 pathway in breast cancer stem-like cells is required for viability and maintenance. AB - Side-population (SP) cells within cancers and cell lines are rare cell populations known to enrich cancer stem-like cells. In this study, we characterized SP cells from the human breast cancer cell line MCF7 as a model for cancer stem-like cells. Compared with non-SP cells, MCF7 SP cells had higher colony-formation ability in vitro and greater tumorigenicity in vivo, suggesting that MCF7 SP cells enrich cancer stem-like cells. cDNA microarray analysis of the SP cells indicated higher expression of ATP-binding cassette transporters and genes involved in quiescence, which were confirmed by quantitative RT-PCR and flow cytometry cell cycle analysis. To identify signal pathways important for cancer stem-like cells, we analyzed cDNA microarray data and identified nine pathways that were altered in the SP cells. To analyze the protein signaling networks, we used reverse-phase signaling pathway protein microarray technology and identified three signaling proteins that are significantly different between MCF7 SP and non-SP cells. Notably, signaling of phosphatidylinositol 3-kinase (PI3K)/mammalian target of rapamycin (mTOR), signal transduction and activator of transcription (STAT3), and phosphatase and tensin homolog (PTEN) was confirmed to be critical for MCF7 SP cell survival and proliferation by pathway specific inhibitors, selected gene knockdown, and in vivo tumorigenicity assay. The STAT3 pathway was found to be positively regulated by mTOR signaling, whereas PTEN served as a negative regulator of both STAT3 and mTOR signaling. This study suggests the existence of prosurvival signaling pathways critical for cancer stem like cell maintenance, which could be selectively targeted for inhibiting cancer stem-like cells for improved treatment. PMID- 17911268 TI - The coordination of cyclic microtubule association/dissociation and tail swing of cytoplasmic dynein. AB - The dynein motor domain is composed of a tail, head, and stalk and is thought to generate a force to microtubules by swinging the tail against the head during its ATPase cycle. For this "power stroke," dynein has to coordinate the tail swing with microtubule association/dissociation at the tip of the stalk. Although a detailed picture of the former process is emerging, the latter process remains to be elucidated. By using the single-headed recombinant motor domain of Dictyostelium cytoplasmic dynein, we address the questions of how the interaction of the motor domain with a microtubule is modulated by ATPase steps, how the two mechanical cycles (the microtubule association/dissociation and tail swing) are coordinated, and which ATPase site among the multiple sites in the motor domain regulates the coordination. Based on steady-state and pre-steady-state measurements, we demonstrate that the two mechanical cycles proceed synchronously at most of the intermediate states in the ATPase cycle: the motor domain in the poststroke state binds strongly to the microtubule with a K(d) of approximately 0.2 microM, whereas most of the motor domains in the prestroke state bind weakly to the microtubule with a K(d) of >10 microM. However, our results suggest that the timings of the microtubule affinity change and tail swing are staggered at the recovery stroke step in which the tail swings from the poststroke to the prestroke position. The ATPase site in the AAA1 module of the motor domain was found to be responsible for the coordination of these two mechanical processes. PMID- 17911269 TI - Spontaneous knotting of an agitated string. AB - It is well known that a jostled string tends to become knotted; yet the factors governing the "spontaneous" formation of various knots are unclear. We performed experiments in which a string was tumbled inside a box and found that complex knots often form within seconds. We used mathematical knot theory to analyze the knots. Above a critical string length, the probability P of knotting at first increased sharply with length but then saturated below 100%. This behavior differs from that of mathematical self-avoiding random walks, where P has been proven to approach 100%. Finite agitation time and jamming of the string due to its stiffness result in lower probability, but P approaches 100% with long, flexible strings. We analyzed the knots by calculating their Jones polynomials via computer analysis of digital photos of the string. Remarkably, almost all were identified as prime knots: 120 different types, having minimum crossing numbers up to 11, were observed in 3,415 trials. All prime knots with up to seven crossings were observed. The relative probability of forming a knot decreased exponentially with minimum crossing number and Mobius energy, mathematical measures of knot complexity. Based on the observation that long, stiff strings tend to form a coiled structure when confined, we propose a simple model to describe the knot formation based on random "braid moves" of the string end. Our model can qualitatively account for the observed distribution of knots and dependence on agitation time and string length. PMID- 17911270 TI - Determinants of mobility and self-care in older people with stroke: importance of somatosensory and perceptual functions. AB - BACKGROUND AND PURPOSE: Somatosensory as well as mental impairments are easily overlooked after acute stroke. Furthermore, their associations with activity limitations are not fully understood. The purpose of this study was to examine this association and whether the assessment of somatosensory functions will provide information on perceptual functions after acute stroke. SUBJECTS AND METHODS: In 115 subjects who were > or =65 years of age, the following parameters were assessed 5 days after stroke: somatosensory (touch and proprioceptive), perceptual, and cognitive functions; depressive symptoms; mobility; and self care. RESULTS: Multivariate analyses showed that normal proprioceptive function was significantly associated with better mobility. Normal perceptual and touch functions were significantly associated with better self-care. Subjects with normal proprioceptive function were 8.6 times as likely to have normal perceptual function as subjects with proprioceptive impairment. DISCUSSION AND CONCLUSION: Somatosensory and perceptual functions were significantly associated with subjects' activity levels. Normal proprioceptive function also might indicate normal perceptual function. PMID- 17911271 TI - Predictors of short-term outcome in people with a clinical diagnosis of cervical radiculopathy. AB - BACKGROUND AND PURPOSE: The purpose of this prospective cohort study was to identify whether variables from the baseline examination or physical therapy interventions received could predict clinical outcomes for people with cervical radiculopathy. SUBJECTS AND METHODS: A total of 96 consecutive patients referred for physical therapy for cervical radiculopathy were the sources of data for this study. All subjects underwent a standardized examination and completed the Neck Disability Index (NDI), the Patient-Specific Functional Scale (PSFS), and the Numeric Pain Rating Scale (NPRS) at baseline and at discharge. The subjects were treated according to the discretion of the individual therapists. At the time of discharge, the subjects completed the Global Rating of Change as well. Subjects surpassing the minimal clinically important change for all 4 outcome tools were categorized as achieving short-term success. Individual variables from the examination and interventions provided were tested for univariate relationships with outcomes. Variables with a significance level of less than .10 were retained as potential predictor variables and were entered into a stepwise logistic regression model to determine the most accurate set of variables for predicting outcomes. RESULTS: The pretest probability for the likelihood of short-term (28 day follow-up) success was 53%. A 4-variable model optimally identified subjects who were most likely to achieve success with physical therapy interventions (age of <54 years; dominant arm is not affected; looking down does not worsen symptoms; and multimodal treatment including manual therapy, cervical traction, and deep neck flexor muscle strengthening for at least 50% of visits). When 3 of these 4 variables were present, the positive likelihood ratio (+LR) was 5.2 (95% confidence interval [CI]=2.4, 11.3), and the posttest probability of success was 85%. When all 4 variables were present, the +LR was 8.3 (95% CI=1.9, 63.9), and the posttest probability of success was 90%. DISCUSSION AND CONCLUSION: These results suggest that a subset of predictor variables can accurately identify which people with cervical radiculopathy are likely to experience short-term successful outcomes. The study design did not allow for the identification of a cause-and-effect relationship, but it appears that intermittent cervical traction, manual therapy, and deep neck flexor muscle strengthening may be beneficial in the management of cervical radiculopathy. Future research is needed to substantiate these findings. PMID- 17911272 TI - Presentation and progression of Friedreich ataxia and implications for physical therapist examination. AB - Friedreich ataxia, although rare, is the most prevalent inherited ataxia. Recent insight into the disease pathogenesis is creating new hope for effective therapies. The purposes of this update are: (1) to review the etiology, presentation, and progression of Friedreich ataxia and (2) to describe a comprehensive physical therapist examination emphasizing valid and reliable performance measurements associated with disease progression. Early identification of individuals with Friedreich ataxia and precise characterization of impairments and functional limitations gain importance as new drug therapies are considered. PMID- 17911273 TI - Determining the resting position of the glenohumeral joint in subjects who are healthy. AB - BACKGROUND AND PURPOSE: The resting position is frequently used by clinicians in the examination and early treatment of patients with joint impairments. However, there is a lack of research on the kinematic characteristics of the resting position of the glenohumeral (GH) joint. The aim of this study was to define the resting position of the GH joint by quantifying the humeral head translation and axial rotational range of motion (ROM). SUBJECTS AND METHODS: The anterior and posterior translation of the humeral head and the rotational ROM of the dominant arm were assessed in the seated position at multiple abduction positions in 15 subjects who were healthy by use of an electromagnetic tracking device. A force of 80 N and a torque of 4 N.m were applied during the measurement procedures for the translation of the humeral head and the rotational ROM, respectively. RESULTS: The mean resting position determined by rotational movement was located at 49.8 degrees of GH abduction. However, the mean resting position determined by translational movement was located at 23.7 degrees of GH abduction and was significantly lower than the resting position determined by rotational movement (t=5.45, P=.000). DISCUSSION AND CONCLUSION: The mean resting position for rotational movement is consistent with the already accepted range of 30 to 60 degrees for a "loosely packed" position of the GH joint. The mean resting position for translational movement appears to be lower than 30 to 60 degrees. The results of this study suggest that, at least for the GH joint, different resting positions should be assessed with different movement criteria (accessory or physiological movement). PMID- 17911274 TI - Number of pedometer-assessed steps taken per day by adults: a descriptive meta analysis. AB - BACKGROUND AND PURPOSE: Although a criterion reference of 10,000 steps per day is recommended for health benefits, a literature search yielded no normative standard for daily steps. Therefore, a meta-analysis was used to estimate the number of pedometer-documented steps that adults take daily. METHODS: Studies were retrieved using electronic databases and cross-referencing from retrieved studies. A random-effects model and 95% confidence intervals (CIs) were used for all analyses of daily step data. RESULTS: Seventy-one outcomes, representing 6,199 subjects from 42 studies, met the inclusion criteria. Excluding Amish subjects, the daily step count averaged 9,448 (95% CI=8,899-9,996) and was greater in subjects younger than 65 years of age (mean=9,797, 95% CI=9,216 10,377) compared with subjects 65 years of age or older (mean=6,565, 95% CI=4,897 8,233). DISCUSSION AND CONCLUSION: This study provides an estimate of the number of daily steps taken by adults. The number is less than the recommended 10,000 steps per day and is especially low in adults 65 years of age or older. PMID- 17911275 TI - CT scans on children: is this a problem? PMID- 17911276 TI - Attention deficit/hyperactivity disorder: a case study in differential diagnosis. PMID- 17911277 TI - Henoch-Schonlein purpura. PMID- 17911278 TI - Fear of animals. PMID- 17911279 TI - Notification that new names and new combinations have appeared in volume 57, part 7, of the IJSEM. AB - This listing of names published in a previous issue of the IJSEM is provided as a service to bacteriology to assist in the recognition of new names and new combinations. This procedure was proposed by the Judicial Commission [Minute 11(ii), Int J Syst Bacteriol 41 (1991), p. 185]. The names given herein are listed according to the Rules of priority (i.e. page number and order of valid publication of names in the original articles). Taxonomic opinions included in this List (i.e. the creation of synonyms or the emendation of circumscriptions) cannot be considered as validly published nor, in any other way, approved by the International Committee on Systematics of Prokaryotes and its Judicial Commission. PMID- 17911280 TI - Thermoanaerobacter pseudethanolicus sp. nov., a thermophilic heterotrophic anaerobe from Yellowstone National Park. AB - Strain 39E(T), originally characterized as Clostridium thermohydrosulfuricum strain 39E and later renamed as Thermoanaerobacter ethanolicus strain 39E, shows less than 97 % 16S rRNA gene sequence similarity with the type strain of the type species of the genus Thermoanaerobacter, T. ethanolicus strain JW 200(T). On the basis of a polyphasic analysis that included DNA-DNA hybridization studies with the subspecies of Thermoanaerobacter brockii, its closest phylogenetic relatives, strain 39E(T) represents a novel species of the genus Thermoanaerobacter, for which the name Thermoanaerobacter pseudethanolicus sp. nov. is proposed. The type strain is 39E(T) (=DSM 2355(T)=ATCC 33223(T)). PMID- 17911281 TI - Halomonas kribbensis sp. nov., a novel moderately halophilic bacterium isolated from a solar saltern in Korea. AB - A moderately halophilic, Gram-negative bacterium, designated strain BH843(T), was isolated from a solar saltern in Korea and subjected to a taxonomic analysis. Strain BH843(T) grew at salinities of 1-14 % (w/v) NaCl and at temperatures of 10 40 degrees C. The cells were motile cocci or short rods with single flagella and contained C(16 : 0), C(19 : 0) cyclo omega8c and C(17 : 0) cyclo as the major fatty acids. The G+C content of the genomic DNA was 66 mol% and the predominant ubiquinone was Q-9. Comparative 16S rRNA gene sequence analyses showed that strain BH843(T) formed a distinct phyletic line within the genus Halomonas, and the levels of 16S rRNA gene sequence similarity with respect to recognized Halomonas species were below 95.1 %. The levels of DNA-DNA relatedness between strain BH843(T) and the type strains of phylogenetically closely related Halomonas species were below 25 %. On the basis of phenotypic, chemotaxonomic and molecular data, strain BH843(T) represents a novel species within the genus Halomonas, for which the name Halomonas kribbensis is proposed. The type strain is BH843(T) (=KCTC 12584(T)=DSM 17892(T)). PMID- 17911282 TI - Halococcus thailandensis sp. nov., from fish sauce in Thailand. AB - Fifteen strains of red-pigmented, strictly aerobic, coccoid, extremely halophilic archaea were isolated from fish sauce (nam-pla) produced in Thailand. They grew optimally at 37 degrees C, pH 6-8 and in the presence of 20-30 % (w/v) NaCl. The DNA G+C contents of the isolates were 60.0-61.8 mol%. They had MK-8(H2) as a major menaquinone component and C(20)C(20) and C(20)C(25) derivatives of phosphatidylglycerol, phosphatidylglycerol methylphosphate and a sulfated glycolipid, S-DGA-1, as major polar lipid components. 16S rRNA gene sequence comparisons revealed that a representative strain, HDB5-2(T), was affiliated with Halococcus dombrowskii JCM 12289(T), Halococcus qingdaonensis JCM 13587(T) and Halococcus morrhuae JCM 8876(T) (levels of similarity of 98.2-98.7 %). Based on data from DNA-DNA hybridization experiments, the 15 strains represented a single species, showing hybridization values of >78.9 % to representative strain HDB5 2(T), but were unrelated to either Halococcus dombrowskii JCM 12289(T) or Halococcus morrhuae JCM 8876(T), with levels of relatedness of <50 %. Moreover, a comparison of phenotypic properties discriminated these new isolates from recognized species of the genus Halococcus. The 15 strains are thus considered to represent a novel species of the genus Halococcus, for which the name Halococcus thailandensis sp. nov. is proposed. The type strain is HDB5-2(T) (=BCC 20213(T) =JCM 13552(T) =PCU 278(T)). PMID- 17911283 TI - Halorubrum litoreum sp. nov., an extremely halophilic archaeon from a solar saltern. AB - An extremely halophilic archaeon, strain Fa-1(T), was isolated from a marine solar saltern in Fujian, China. Strain Fa-1(T) required Mg2+ and at least 2.0 M NaCl for growth. It was able to grow at pH 6.5-9.0 (optimally at pH 7.0-7.5) and at 20-55 degrees C (optimally at 37-42 degrees C). The major polar lipids of strain Fa-1(T) were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and a sulfated diglycosyl diether. On the basis of a 16S rRNA gene sequence analysis, strain Fa-1(T) was closely related to nine species of the genus Halorubrum, showing sequence similarities of 97.4-98.4 %. The G+C content of the DNA of strain Fa-1(T) is 64.9 mol% (Tm). DNA-DNA hybridization values between strain Fa-1(T) and the most closely related members of the genus Halorubrum were below 51 %. On the basis of the data from this study, strain Fa-1(T) represents a novel species of the genus Halorubrum, for which the name Halorubrum litoreum sp. nov. is proposed. The type strain is Fa 1(T) (=CGMCC 1.5336(T) =JCM 13561(T)). PMID- 17911284 TI - Altererythrobacter epoxidivorans gen. nov., sp. nov., an epoxide hydrolase active, mesophilic marine bacterium isolated from cold-seep sediment, and reclassification of Erythrobacter luteolus Yoon et al. 2005 as Altererythrobacter luteolus comb. nov. AB - A novel marine bacterium, strain JCS350(T), was isolated from marine sediment samples collected from a cold-seep area. The 16S rRNA gene sequence of the isolate showed high similarity to that of Erythrobacter luteolus SW-109(T) (95.9 % sequence similarity). Lower 16S rRNA gene sequence similarities were shown to other members of the genus Erythrobacter (94.6-95.4 %) and members of the genus Porphyrobacter (94.5-95.2 %). Phylogenetic analysis with all members of the family Erythrobacteraceae and several members of the family Sphingomonadaceae revealed that the isolate formed a phyletic line with [Erythrobacter] luteolus that was distinct from other members of the family Erythrobacteraceae. The dominant fatty acids of strain JCS350(T) were 18 : 1omega7c, 16 : 1omega7c and cyclopropane 17 : 0. The major respiratory quinone was ubiquinone 10. The DNA G+C content was 54.5 mol%. The isolate did not contain bacteriochlorophyll a. Optimal growth required the presence of 2 % (w/v) NaCl with either 0.18 % CaCl(2) or 0.59 % MgCl(2), at pH 6.5 and at 35 degrees C. On the basis of the evidence of this polyphasic taxonomic study, strain JCS350(T) should be classified in a novel genus and species in the family Erythrobacteraceae, for which the name Altererythrobacter epoxidivorans gen. nov., sp. nov. is proposed. The misclassified species [Erythrobacter] luteolus is transferred to the new genus as Altererythrobacter luteolus comb. nov. The type strain of Altererythrobacter epoxidivorans is JCS350(T) (=KCCM 42314(T) =JCM 13815(T)) and the type strain of Altererythrobacter luteolus is SW-109(T) (=KCTC 12311(T) =JCM 12599(T)). PMID- 17911285 TI - Devosia subaequoris sp. nov., isolated from beach sediment. AB - A novel marine bacterium was isolated from a sediment sample from Hwasun Beach in Jeju, Republic of Korea. The cells were found to be Gram-negative, aerobic, oxidase-positive, catalase-positive, motile rods. The organism required natural seawater or artificial sea salts for growth. The temperature and pH ranges for growth were 20-42 degrees C and pH 5.1-12.1, respectively. Phylogenetic analyses based on 16S rRNA gene sequences showed that the organism belonged to the order Rhizobiales and formed a robust cluster with members of the genus Devosia. Its phylogenetic neighbours were the type strains of Devosia riboflavina (96.8 % 16S rRNA gene sequence similarity), Devosia neptuniae (96.7 %), Devosia soli (96.5 %) and Devosia limi (96.2 %), 'Devosia terrae' DCY11 (96.2 %) and 'Candidatus Devosia euplotis' (96.2 %). The predominant ubiquinone was Q-10, the major fatty acids were C(18 : 1), C(18 : 0) and C(16 : 0) and the G+C content of the DNA was 59.1 mol%. On the basis of phenotypic data and the results of phylogenetic analyses, strain HST3-14(T) represents a novel species of the genus Devosia, for which the name Devosia subaequoris sp. nov. is proposed. The type strain is strain HST3-14(T) (=KCTC 12772(T) =JCM 14206(T)). PMID- 17911286 TI - Prevotella nanceiensis sp. nov., isolated from human clinical samples. AB - Three strains of anaerobic, non-pigmented, Gram-negative bacilli isolated from various human clinical samples were characterized in terms of phenotypic and genotypic tests, including sequence analysis of 16S rRNA and rpoB genes. The strains were most closely related to the type strains of Prevotella marshii and Prevotella shahii on the basis of both 16S rRNA (89.8 and 89.0 % identity, respectively) and rpoB gene sequences (83.1 and 82.8 % identity, respectively). Phylogenetic analysis showed that the isolates constituted a robust homogeneous group distinct from known species in the genus Prevotella. The rrn skeleton (as determined by PFGE) and the DNA G+C content, determined to be 39.4 mol% for strain LBN 293(T), distinguished the novel isolates from the type strains of P. marshii and P. shahii. The three strains were saccharolytic and produced acetic, lactic and succinic acids as major metabolic end products. Polyphasic investigations supported the proposal of a novel species, Prevotella nanceiensis sp. nov., with LBN 293(T) (=AIP 261.03(T) =CIP 108993(T) =CCUG 54409(T)) as the type strain. PMID- 17911287 TI - Pedobacter duraquae sp. nov., Pedobacter westerhofensis sp. nov., Pedobacter metabolipauper sp. nov., Pedobacter hartonius sp. nov. and Pedobacter steynii sp. nov., isolated from a hard-water rivulet. AB - Five isolates that were related phylogenetically to members of the genus Pedobacter were isolated from freshwater of the hard-water creek Westerhofer Bach, North Germany. The five strains (WB 2.1-25(T), WB 2.3-71(T), WB 3.3-3(T), WB 3.3-22(T) and WB 2.3-45(T)) were Gram-negative and chemoheterotrophic, with rod-shaped cells. Most of their metabolic properties matched those given in the description of the genus Pedobacter. Consistent with the genus description, their fatty acids included mainly iso-C(15 : 0) and summed feature 3 (C(16 : 1)omega7c, iso-C(15 : 0) 2-OH or both); C(16 : 1)omega5c, C(16 : 0), iso-C(15 : 0) 3-OH, C(16 : 0) 3-OH and iso-C(17 : 0) 3-OH were present in smaller amounts. The major isoprenoid quinone was menaquinone 7. With one exception, binary similarity values of the almost complete 16S rRNA gene sequences determined among the isolates as well as between the isolates and type strains of Pedobacter species were lower than 98.5 %. The only exception was the close relationship between Pedobacter caeni DSM 16990(T) and strain WB 2.3-45(T) (99.2 % similarity). DNA DNA reassociation values determined for this pair of strains was 29.8 %, indicating that strain WB 2.3-45(T) represents a unique genospecies. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains WB 2.1-25(T) and WB 2.3-71(T) form a group that is moderately related to P. caeni and strain WB 2.3-45(T) (98.5 % similarity). Strains WB 3.3-3(T) and WB 3.3-22(T) (98.5 % similarity) branched separately from these four organisms. The five phylogenetically isolated strains differed from each other as well as from the type strain of the type species (Pedobacter heparinus DSM 2366(T)) and some related representatives of the genus in several metabolic reactions and cultural parameters. On the basis of phenotypic and phylogenetic distinctiveness, five novel species are proposed: Pedobacter duraquae sp. nov., with WB 2.1-25(T) (=DSM 19034(T)=CIP 109481(T)) as the type strain; Pedobacter westerhofensis sp. nov., with WB 3.3-22(T) (=DSM 19036(T)=CIP 109479(T)) as the type strain; Pedobacter metabolipauper sp. nov., with WB 2.3-71(T) (=DSM 19035(T)=CIP 109480(T)) as the type strain; Pedobacter hartonius sp. nov., with WB 3.3-3(T) (=DSM 19033(T)=CIP 109468(T)) as the type strain; and Pedobacter steynii sp. nov., with WB 2.3-45(T) (=DSM 19110(T)=CIP 109507(T)) as the type strain. PMID- 17911288 TI - Burkholderia bryophila sp. nov. and Burkholderia megapolitana sp. nov., moss associated species with antifungal and plant-growth-promoting properties. AB - A polyphasic taxonomic study including DNA-DNA reassociation experiments and an extensive biochemical characterization was performed on 14 Burkholderia isolates from moss gametophytes of nutrient-poor plant communities on the southern Baltic Sea coast in northern Germany. The strains were classified within two novel species, for which the names Burkholderia bryophila sp. nov. and Burkholderia megapolitana sp. nov. are proposed. The former species also includes isolates from grassland and agricultural soil collected in previous studies. Strains Burkholderia bryophila 1S18(T) (=LMG 23644(T) =CCUG 52993(T)) and Burkholderia megapolitana A3(T) (=LMG 23650(T) =CCUG 53006(T)) are the proposed type strains. They were isolated from Sphagnum rubellum and Aulacomnium palustre, respectively, growing in the 'Ribnitzer Grosses Moor' nature reserve (Mecklenburg-Pommern, Germany). All moss isolates of both novel species showed antifungal activity against phytopathogens as well as plant-growth-promoting properties. PMID- 17911289 TI - Brevundimonas lenta sp. nov., isolated from soil. AB - A Gram-negative, rod-shaped, Brevundimonas-like bacterial strain, DS-18(T), was isolated from soil in Dokdo, Korea, and its exact taxonomic position was investigated by using a polyphasic approach. Strain DS-18(T) grew optimally at pH 6.5-7.0 and 25 degrees C without NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain DS-18(T) belonged to the genus Brevundimonas. Strain DS-18(T) contained Q-10 as the predominant ubiquinone and C(18 : 1)omega7c and C(16 : 0) as the major fatty acids. The DNA G+C content was 68.7 mol%. Strain DS-18(T) exhibited levels of 16S rRNA gene sequence similarity of 96.3-98.7 % to the type strains of Brevundimonas species and Mycoplana bullata. Mean DNA-DNA relatedness values between strain DS-18(T) and the type strains of phylogenetically related Brevundimonas species and M. bullata were in the range 15-32 %. Strain DS-18(T) differed from Brevundimonas species and M. bullata in several phenotypic characteristics. On the basis of phenotypic, phylogenetic and genetic data, strain DS-18(T) represents a novel species of the genus Brevundimonas, for which the name Brevundimonas lenta sp. nov. is proposed. The type strain is DS-18(T) (=KCTC 12871(T) =JCM 14602(T)). PMID- 17911290 TI - Vibrio rhizosphaerae sp. nov., a red-pigmented bacterium that antagonizes phytopathogenic bacteria. AB - Two novel red-pigmented Vibrio strains, MSSRF3(T) and MSSRF10, with antibacterial activity against phytopathogens were isolated from the rhizosphere region of mangrove-associated wild rice (Porteresia coarctata Tateoka), in Pichavaram, India. The cells were Gram-negative, facultatively anaerobic and rod-shaped and were motile by means of single polar flagella. The two strains were catalase positive and oxidase-negative, and were able to grow in 0.1-10 % NaCl (with optimum growth in 2 % NaCl) and at temperatures of 20-42 degrees C (optimum growth at 25-30 degrees C). Both strains produced acid and gas from D-glucose under anaerobic conditions and utilized a wide range of compounds as sole carbon and energy sources. The DNA G+C contents determined were 51.3 mol% for strain MSSRF3(T) and 51.0 mol% for strain MSSRF10. Phylogenetic analysis based on 16S rRNA, rpoA, recA and pyrH gene sequences showed that strains MSSRF3(T) and MSSRF10 belong to the genus Vibrio and are very closely related to Vibrio ruber JCM 11486(T), with which they share 98.3-98.5 % (16S rRNA), 98.3-99.7 % (rpoA), 90.2-99.8 % (recA) and 91.3-99.4 % (pyrH) gene sequence similarities, respectively. Levels of DNA-DNA relatedness were 44 % between strains MSSRF3(T) and MSSRF10, 80 % between strain MSSRF10 and V. ruber JCM 11486(T) and 45 % between strain MSSRF3(T) and V. ruber JCM 11486(T). Strain MSSRF3(T) was phenotypically similar to V. ruber JCM 11486(T). However, the inability to reduce nitrate to nitrite, the ability to grow in 0.1 % NaCl and the presence of caseinase were characteristics that allowed differentiation between V. ruber JCM 11486(T) and strain MSSRF3(T). In addition, strain MSSRF3(T) could be differentiated from strain MSSRF10 and its closest relative V. ruber JCM 11486(T) with respect to its genomic fingerprinting analysis (random amplified polymorphic DNA, GTG5, BOX, PCR-restriction fragment length polymorphism and ribotyping). Therefore, based on phenotypic, genotypic, phylogenetic and DNA-DNA hybridization analyses, strain MSSRF3(T) (=LMG 23790(T)=DSM 18581(T)) should be classified as representing the type strain of a novel species of the genus Vibrio, for which the name Vibrio rhizosphaerae sp. nov. is proposed. PMID- 17911292 TI - EzTaxon: a web-based tool for the identification of prokaryotes based on 16S ribosomal RNA gene sequences. AB - 16S rRNA gene sequences have been widely used for the identification of prokaryotes. However, the flood of sequences of non-type strains and the lack of a peer-reviewed database for 16S rRNA gene sequences of type strains have made routine identification of isolates difficult and labour-intensive. In the present study, we generated a database containing 16S rRNA gene sequences of all prokaryotic type strains. In addition, a web-based tool, named EzTaxon, for analysis of 16S rRNA gene sequences was constructed to achieve identification of isolates based on pairwise nucleotide similarity values and phylogenetic inference methods. The system developed provides users with a similarity-based search, multiple sequence alignment and various phylogenetic analyses. All of these functions together with the 16S rRNA gene sequence database of type strains can be successfully used for automated and reliable identification of prokaryotic isolates. The EzTaxon server is freely accessible over the Internet at http://www.eztaxon.org/ PMID- 17911291 TI - Phylogeny of the Mycoplasma mycoides cluster based on analysis of five conserved protein-coding sequences and possible implications for the taxonomy of the group. AB - A phylogenetic tree of the Mycoplasma mycoides cluster was inferred from a set of concatenated sequences from five housekeeping genes (fusA, glpQ, gyrB, lepA and rpoB). The relevance of this phylogeny was reinforced by detailed analysis of the congruence of the phylogenies derived from each of the five individual gene sequences. Two subclusters were distinguished. The M. mycoides subcluster comprised M. mycoides subsp. mycoides biotypes Small Colony (SC) and Large Colony (LC) and M. mycoides subsp. capri. The latter two groups could not be clearly separated, which supports previous proposals that they be united into a single taxonomic entity. The Mycoplasma capricolum subcluster included M. capricolum subsp. capricolum, M. capricolum subsp. capripneumoniae and Mycoplasma sp. bovine group 7 of Leach, a group of strains that remains unassigned. This group constituted a distinct branch within this cluster, supporting its classification as a subspecies of M. capricolum. Mycoplasma cottewii and Mycoplasma yeatsii clustered in a group that was distinct from Mycoplasma putrefaciens and they were all clearly separated from the M. mycoides cluster. In conclusion, this approach has allowed us to assign phylogenetic positions to all members of the M. mycoides cluster and related species and has proved the need to adjust the existing taxonomy. Furthermore, this method may be used as a reference technique to assign an unequivocal position to any particular strain related to this cluster and may lead to the development of new techniques for rapid species identification. PMID- 17911293 TI - Chromohalobacter japonicus sp. nov., a moderately halophilic bacterium isolated from a Japanese salty food. AB - A Gram-negative, non-spore-forming, rod-shaped, motile bacterium, designated strain 43(T), was isolated from a Japanese salty food and then subjected to a polyphasic taxonomic study. Strain 43(T) is moderately halophilic, growing at NaCl concentrations in the range 5-25 % (w/v), with optimum growth between 7.5 and 12.5 % (w/v) NaCl. Growth occurs at temperatures from 15 to 42 degrees C (optimally at 28-37 degrees C) and at pH 5.5-9.0 (optimally at pH 7.0-8.0). A phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain 43(T) belongs to the genus Chromohalobacter. The closest relatives were Chromohalobacter canadensis ATCC 43984(T) (99.3 % 16S rRNA gene sequence similarity), Chromohalobacter beijerinckii ATCC 19372(T) (99.1 %), Chromohalobacter sarecensis LV4(T) (98.3 %), Chromohalobacter nigrandesensis LTS 4N(T) (97.9 %) and Chromohalobacter marismortui ATCC 17056(T) (97.9 %). The DNA G+C content was 62.9 mol%, which is within the range described for the genus Chromohalobacter. DNA-DNA hybridization studies between strain 43(T) and C. canadensis CECT 5385(T) and C. beijerinckii DSM 7218(T) showed 38 and 49 % relatedness, respectively; lower DNA-DNA hybridization percentages were obtained with respect to other related Chromohalobacter species. The major fatty acids of strain 43(T) were C(16 : 0), C(19 : 0) cyclo omega8c and C(12 : 0) 3-OH. Overall, the phenotypic, genotypic and phylogenetic results demonstrated that strain 43(T) represents a novel species within the genus Chromohalobacter. The name Chromohalobacter japonicus sp. nov. is proposed, with strain 43(T) (=CECT 7219(T) =CCM 7416(T)) as the type strain. PMID- 17911294 TI - Paenibacillus humicus sp. nov., isolated from poultry litter compost. AB - Two bacterial strains, PC-142 and PC-147(T), isolated from poultry litter compost, were characterized with respect to their phenetic and phylogenetic characteristics. The isolates were endospore-forming rods that were reddish in colour after Gram staining. They were catalase- and oxidase-positive, were able to degrade starch and gelatin and grew at 15-40 degrees C and pH 5.5-10.0. The predominant fatty acids were anteiso-C(15 : 0), iso-C(15 : 0) and iso-C(16 : 0), the major respiratory quinone was menaquinone MK-7, the cell-wall peptidoglycan was of the A1gamma type and the G+C content of the DNA was 58 mol%. The 16S rRNA gene sequence analysis and phenetic characterization indicated that these organisms belong to the genus Paenibacillus, with Paenibacillus pasadenensis SAFN 007(T) as the closest phylogenetic neighbour (97.5 %). Strains PC-142, PC-147(T) and P. pasadenensis SAFN-007(T) represent a novel lineage within the genus Paenibacillus, characterized by a high DNA G+C content (58-63 mol%). The low levels of 16S rRNA gene sequence similarity with respect to other taxa with validly published names and the identification of distinctive phenetic features in the two isolates indicate that strains PC-142 and PC-147(T) represent a novel species of the genus Paenibacillus, for which the name Paenibacillus humicus sp. nov. is proposed. The type strain is PC-147(T) (=DSM 18784(T) =NBRC 102415(T) =LMG 23886(T)). PMID- 17911295 TI - Catenulispora rubra sp. nov., an acidophilic actinomycete isolated from forest soil. AB - In the course of screening novel secondary metabolites, an acidophilic actinomycete strain, designated Aac-30(T), was isolated from forest soil and subjected to a polyphasic taxonomic characterization study. It grew well on media in which the pH ranged from 4 to 6, but not on medium with pH adjusted to 7. It possessed ll-diaminopimelic acid and glycine in the cell-wall peptidoglycan, MK 9(H6) and MK-9(H8) as major isoprenoid quinones, iso-C(16 : 0) and anteiso-C(17 : 0) as major cellular fatty acids, and phosphatidylglycerol and phosphatidylinositol as polar lipids. The G+C content of the genomic DNA was 69.1 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that the organism belonged to the family Catenulisporaceae and consistently formed a monophyletic cluster with members of the genus Catenulispora. Physiological, biochemical and chemotaxonomic data revealed that this novel organism could be readily differentiated from recognized members of the genus Catenulispora and that it merits separate species status. Based on the phenotypic and genetic evidence presented, strain Aac-30(T) is considered to represent a novel species of the genus Catenulispora, for which the name Catenulispora rubra sp. nov. is proposed. The type strain is Aac-30(T) (=NBRC 101179(T)=DSM 44948(T)). PMID- 17911296 TI - Rapidithrix thailandica gen. nov., sp. nov., a marine gliding bacterium isolated from samples collected from the Andaman sea, along the southern coastline of Thailand. AB - The taxonomic positions of three strains of marine gliding bacteria, TISTR 1736, TISTR 1741 and TISTR 1750(T), isolated from the southern coastline of Thailand were evaluated by using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the three isolates formed a distinct lineage within the family 'Flammeovirgaceae', phylum Bacteroidetes, and were related to the genus Flexithrix. The DNA G+C contents of the isolates were in the range 40 43 mol%. The major respiratory quinone was MK-7. The major cellular fatty acids were 16 : 1omega5c (cis-5-hexadecenoic acid) and 15 : 0 (pentadecanoic acid). The major hydroxyl fatty acids were 3-OH 17 : 0 (3-hydroxyheptadecanoic acid), 3-OH 15 : 0 (3-hydroxypentadecanoic acid) and 3-OH 16 : 0 (3-hydroxyhexadecanoic acid). On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic data, these marine bacteria are considered to represent a novel species of a new genus, for which the name Rapidithrix thailandica gen. nov., sp. nov. is proposed. The type strain of Rapidithrix thailandica is TISTR 1750(T) (=IAM 15448(T)). PMID- 17911297 TI - Kaistia granuli sp. nov., isolated from anaerobic granules in an upflow anaerobic sludge blanket reactor. AB - A Gram-negative, chemo-organotrophic, non-motile, non-spore-forming, rod-shaped bacterium (designated strain Ko04(T)) was isolated from anaerobic granules in an upflow anaerobic sludge blanket reactor, and was investigated using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Ko04(T) belongs to the order Rhizobiales in the Alphaproteobacteria. Comparative 16S rRNA gene sequence analysis showed that strain Ko04(T) was most closely related to Kaistia adipata (97.5 %) and that sequence similarities with other species of Rhizobiales with validly published names were less than 92.5 %. The predominant ubiquinone was Q-10 and the major fatty acids were C(18 : 1)omega7c/omega9t/omega12t, C(19 : 0 )cyclo omega8c and C(18 : 0). The G+C content of the genomic DNA of strain Ko04(T) was 67.8 mol%. The level of DNA-DNA relatedness with K. adipata Chj404(T) was 15 %. The results of the genotypic analyses in combination with chemotaxonomic and physiological data demonstrated that strain Ko04(T) represents a novel species within the genus Kaistia, for which the name Kaistia granuli sp. nov. is proposed. The type strain is Ko04(T) (=KCTC 12575(T)=LMG 23410(T)). PMID- 17911298 TI - Herbaspirillum rhizosphaerae sp. nov., isolated from rhizosphere soil of Allium victorialis var. platyphyllum. AB - Two Gram-negative, milky-white-pigmented, motile, slightly curved rod-shaped bacterial isolates, UMS-37(T) and UMS-40, were isolated from rhizosphere soil of wild edible greens cultivated on Ulleung island, Korea, and their taxonomic positions were investigated by a polyphasic approach. They grew optimally at 25 30 degrees C and contained Q-8 as the predominant ubiquinone. The major cellular fatty acids (>10 % of total fatty acids) were C(16 : 0), cyclo C(17 : 0) and C(16 : 1)omega7c and/oriso-C(15 : 0) 2-OH. The DNA G+C contents of the two isolates were 59.8 and 60.0 mol%. Isolates UMS-37(T) and UMS-40 exhibited no difference in their 16S rRNA gene sequences and possessed a mean DNA-DNA relatedness level of 94 %; they exhibited 16S rRNA gene sequence similarity levels of 96.8-98.2 % to the type strains of recognized Herbaspirillum species. Phylogenetic analyses based on 16S rRNA gene sequences showed that isolates UMS-37(T) and UMS-40 formed a distinct phylogenetic lineage within the genus Herbaspirillum. DNA-DNA relatedness levels between isolates UMS-37(T) and UMS-40 and the type strains of some phylogenetically related Herbaspirillum species were in the range 3-56 %. On the basis of differences in phenotypic properties and phylogenetic distinctiveness and genomic data, isolates UMS-37(T) and UMS-40 were classified in the genus Herbaspirillum within a novel species, for which the name Herbaspirillum rhizosphaerae sp. nov. is proposed, with the type strain UMS-37(T) (=KCTC 12558(T) =CIP 108917(T)). PMID- 17911299 TI - Phylogenetic relationships within the family Halobacteriaceae inferred from rpoB' gene and protein sequences. AB - In order to clarify the current phylogeny of the haloarchaea, particularly the closely related genera that have been difficult to sort out using 16S rRNA gene sequences, the DNA-dependent RNA polymerase subunit B' gene (rpoB') was used as a complementary molecular marker. Partial sequences of the gene were determined from 16 strains of the family Halobacteriaceae. Comparisons of phylogenetic trees inferred from the gene and protein sequences as well as from corresponding 16S rRNA gene sequences suggested that species of the genera Natrialba, Natronococcus, Halobiforma, Natronobacterium, Natronorubrum, Natrinema/Haloterrigena and Natronolimnobius formed a monophyletic group in all trees. In the RpoB' protein tree, the alkaliphilic species Natrialba chahannaoensis, Natrialba hulunbeirensis and Natrialba magadii formed a tight group, while the neutrophilic species Natrialba asiatica formed a separate group with species of the genera Natronorubrum and Natronolimnobius. Species of the genus Natronorubrum were split into two groups in both the rpoB' gene and protein trees. The most important advantage of the use of the rpoB' gene over the 16S rRNA gene is that sequences of the former are highly conserved amongst species of the family Halobacteriaceae. All sequences determined so far can be aligned unambiguously without any gaps. On the other hand, gaps are necessary at 49 positions in the inner part of the alignment of 16S rRNA gene sequences. The rpoB' gene and protein sequences can be used as an excellent alternative molecular marker in phylogenetic analysis of the Halobacteriaceae. PMID- 17911300 TI - Halalkalicoccus jeotgali sp. nov., a halophilic archaeon from shrimp jeotgal, a traditional Korean fermented seafood. AB - A novel, extremely halophilic archaeon B3(T) was isolated from shrimp-salted seafood. Its morphology, physiology, biochemical features and 16S rRNA gene sequence were characterized. Strain B3(T) is non-motile, Gram-variable, requires at least 10 % (w/v) NaCl for growth and grows in the ranges of 21-50 degrees C and pH 6.5-9.0. The DNA G+C content of strain B3(T) was 63.2 mol%. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain B3(T) belonged to the genus Halalkalicoccus and was phylogenetically closely related to the type strain Halalkalicoccus tibetensis (98.64 %). However, DNA-DNA hybridization experiments showed 7.0 % relatedness between strain B3(T) and a strain of a reference species of the genus Halalkalicoccus. Combined analysis of 16S rRNA gene sequences, DNA-DNA relatedness data, physiological and biochemical tests indicated that the genotypic and phenotypic characteristics differentiate strain B3(T) from other Halalkalicoccus species. On the basis of the evidence presented in this report, strain B3(T) represents a novel species of the genus Halalkalicoccus, for which the name Halalkalicoccus jeotgali. sp. nov. is proposed. The type strain is B3(T) (=KCTC 4019(T)=DSM 18796(T)=JCM 14584(T)=CECT 7217(T)). PMID- 17911302 TI - Modicisalibacter tunisiensis gen. nov., sp. nov., an aerobic, moderately halophilic bacterium isolated from an oilfield-water injection sample, and emended description of the family Halomonadaceae Franzmann et al. 1989 emend Dobson and Franzmann 1996 emend. Ntougias et al. 2007. AB - An aerobic, moderately halophilic, Gram-negative, motile, non-sporulating bacterium, strain LIT2(T), was isolated from an oilfield-water injection after enrichment on crude oil. Strain LIT2(T) grew between 15 and 45 degrees C and optimally at 37 degrees C. It grew in the presence of 1-25 % (w/v) NaCl, with an optimum at 10 % (w/v) NaCl. Predominant fatty acids were C(16 : 0) (26.9 %), C(18 : 1)omega7c (22.6 %), C(16 : 1)omega7c (20.4 %) C(19 : 0) cyclo omega8c (10.9 %) and C(17 : 0) (8 %). Interestingly, the relative percentages of these last two fatty acids were intermediate compared with most species among the family Halomonadaceae for which fatty acid composition has been determined. The DNA G+C content was 53.7 mol%, which is very low among the family Halomonadaceae. Strain LIT2(T) exhibited 16S rRNA gene sequence similarity values of 94.06-95.15 % to members of the genus Chromohalobacter, 94.21-94.65 % to members of the genus Halomonas and 93.57 % with the single species representative of the genus Cobetia. Based on the phylogenetic and phenotypic evidence presented in this paper, we propose the name Modicisalibacter tunisiensis gen. nov., sp. nov. to accommodate strain LIT2(T). The type strain of Modicisalibacter tunisiensis is LIT2(T) (=CCUG 52917(T) =CIP 109206(T)). A reassignment of the descriptive 16S rRNA signature characteristics of the family Halomonadaceae permitted placement of the new genus Modicisalibacter into the family. PMID- 17911301 TI - Bellilinea caldifistulae gen. nov., sp. nov. and Longilinea arvoryzae gen. nov., sp. nov., strictly anaerobic, filamentous bacteria of the phylum Chloroflexi isolated from methanogenic propionate-degrading consortia. AB - Thermophilic (strain GOMI-1(T)) and mesophilic (strain KOME-1(T)) strains were isolated from two different cultures of propionate-degrading consortia obtained from thermophilic digester sludge and rice paddy soil, respectively. The two strains were non-spore-forming, non-motile and Gram-negative. Both strains were obligately anaerobic micro-organisms, showing multicellular filamentous morphotypes more than 100 mum in length. The cell width for strain GOMI-1(T) was 0.2-0.4 mum and that of strain KOME-1(T) was 0.4-0.6 mum. Strain GOMI-1(T) could grow at 45-65 degrees C with a pH range of 6.0-7.5 (optimum growth at 55 degrees C, pH 7.0). The temperature range for growth of strain KOME-1(T) was 30-40 degrees C and the pH range was pH 5.0-8.5 (optimum growth around 37 degrees C, pH 7.0). Yeast extract was required for growth of both strains. Strain GOMI-1(T) was able to grow with a number of carbohydrates in the presence of yeast extract. In yeast extract-containing medium, strain KOME-1(T) could utilize proteins and a limited range of sugars for growth. The G+C contents of the DNA of strains GOMI 1(T) and KOME-1(T) were respectively 54.7 and 57.6 mol%. Major fatty acids of strain GOMI-1(T) were C(16 : 0), C(14 : 0) and iso-C(15 : 0), whereas those of strain KOME-1(T) were iso-C(15 : 0), anteiso-C(15 : 0) and C(14 : 0). Based on comparative analysis of 16S rRNA gene sequences of strains GOMI-1(T) and KOME 1(T), the strains were placed in different phylogenetic positions in the class Anaerolineae of the bacterial phylum Chloroflexi. Their phenotypic and genetic traits strongly supported the conclusion that the strains should be described as two independent taxa in the class Anaerolineae. Hence, we propose the names Bellilinea caldifistulae gen. nov., sp. nov., and Longilinea arvoryzae gen. nov., sp. nov., for strains GOMI-1(T) and KOME-1(T). The type strains of Bellilinea caldifistulae and Longilinea arvoryzae are respectively GOMI-1(T) (=JCM 13669(T) =DSM 17877(T)) and KOME-1(T) (=JCM 13670(T) =KTCC 5380(T)). PMID- 17911303 TI - Acidaminococcus intestini sp. nov., isolated from human clinical samples. AB - Eleven strains of a hitherto unknown, Gram-negative, anaerobic coccus were recovered from various human clinical samples of patients hospitalized in two geographically distant French hospitals. These strains displayed the morphology and growth characteristics of those related to the genus Acidaminococcus. The clinical isolates shared at least 99.9 and 99.7 % of their nucleotide positions in the 16S and 23S rRNA gene sequences, respectively. They displayed 95.6 and 88.9 % 16S and 23S rRNA gene sequence similarities, respectively, with Acidaminococcus fermentans. The 16S rRNA-based phylogeny revealed that all the clinical isolates grouped in a statistically well supported cluster separate from A. fermentans. Enzymic activity profiles as well as metabolic end product patterns, including propionic acid production, differentiated the novel bacteria from A. fermentans. Finally, phenotypic, genotypic and phylogenetic data, including large-scale chromosome structure and DNA G+C content, supported the proposal of a novel species of the genus Acidaminococcus, for which the name Acidaminococcus intestini sp. nov. is proposed. The type strain is ADV 255.99(T) (=AIP 283.01(T)=CIP 108586(T)=CCUG 50930(T)). PMID- 17911304 TI - Dethiosulfatibacter aminovorans gen. nov., sp. nov., a novel thiosulfate-reducing bacterium isolated from coastal marine sediment via sulfate-reducing enrichment with Casamino acids. AB - A sulfate-reducing enrichment culture originating from coastal marine sediment of the eutrophic Tokyo Bay, Japan, was successfully established with Casamino acids as a substrate. A thiosulfate reducer, strain C/G2(T), was isolated from the enrichment culture after further enrichment with glutamate. Cells of strain C/G2(T) were non-motile rods (0.6-0.8 microm x 2.2-4.8 microm) and were found singly or in pairs and sometimes in short chains. Spores were not formed. Cells of strain C/G2(T) stained Gram-negatively, despite possessing Gram-positive cell walls. The optimum temperature for growth was 28-30 degrees C, the optimum pH was around 7.8 and the optimum salt concentration was 20-30 g l(-1). Lactate, pyruvate, serine, cysteine, threonine, glutamate, histidine, lysine, arginine, Casamino acids, peptone and yeast extract were fermented as single substrates and no sugar was used as a fermentative substrate. A Stickland reaction was observed with some pairs of amino acids. Fumarate, alanine, proline, phenylalanine, tryptophan, glutamine and aspartate were utilized only in the presence of thiosulfate. Strain C/G2(T) fermented glutamate to H2, CO2, acetate and propionate. Thiosulfate and elemental sulfur were reduced to sulfide. Sulfate, sulfite and nitrate were not utilized as electron acceptors. The growth of strain C/G2(T) on Casamino acids or glutamate was enhanced by co-culturing with Desulfovibrio sp. isolated from the original mixed culture enriched with Casamino acids. The DNA G+C content of strain C/G2(T) was 41.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain C/G2(T) formed a distinct cluster with species of the genus Sedimentibacter. The closest relative was Sedimentibacter hydroxybenzoicus (with a gene sequence similarity of 91 %). On the basis of its phylogenetic and phenotypic properties, strain C/G2(T) (=JCM 13356(T)=NBRC 101112(T)=DSM 17477(T)) is proposed as representing a new genus and novel species, Dethiosulfatibacter aminovorans gen. nov., sp. nov. PMID- 17911306 TI - Fervidobacterium changbaicum sp. nov., a novel thermophilic anaerobic bacterium isolated from a hot spring of the Changbai Mountains, China. AB - A thermophilic, obligately anaerobic, rod-shaped bacterium (strain CBS-1(T)) was isolated from a hot spring mixture of water and mud of the Changbai Mountains, China. Strain CBS-1(T) was found to be non-sporulating, Gram-negative, with optimal growth at 75-80 degrees C. It grew on a wide range of carbon sources, including glucose, lactose, maltose, starch, sorbitol and pyruvate amongst others. The DNA G+C content of strain CBS-1(T) was 31.9 mol%. The 16S rRNA gene sequence analysis indicated that the strain was a member of the genus Fervidobacterium. The high concentration of C(16 : 0) (52.2 %) in the fatty acid profile of the cell envelope supported its inclusion as a member of the genus Fervidobacterium. On the basis of the low values of DNA-DNA hybridization (25.8 and 20.5 %) and phenotypic features, strain CBS-1(T) represents a novel species of the genus Fervidobacterium, for which the name Fervidobacterium changbaicum sp. nov. is proposed. The type strain is CBS-1(T) (=DSM 17883(T)=JCM 13353(T)). PMID- 17911305 TI - Salinicoccus kunmingensis sp. nov., a moderately halophilic bacterium isolated from a salt mine in Yunnan, south-west China. AB - A novel yellow-pigmented, non-motile, non-sporulating, catalase- and oxidase positive, obligately aerobic, moderately halophilic, facultatively alkaliphilic Gram-positive coccus, strain YIM Y15(T), was isolated from a brine sample from a salt mine in Yunnan, south-west China. Strain YIM Y15(T) grew in the presence of 0.5-25 % (w/v) NaCl and at pH 6.0-10.0, with optimum growth at 8-10 % (w/v) NaCl and pH 8.0. It grew at 4-45 degrees C, with optimum growth at 37.0 degrees C. The major cellular fatty acids were anteiso-C(15 : 0) and iso-C(15 : 0). Menaquinone 6 (MK-6) was the major respiratory quinone. The cell wall contained Lys and Gly. The DNA G+C content was 46.2 mol%. Phylogenetic analyses based on 16S rRNA gene sequence comparisons revealed that strain YIM Y15(T) was a member of the genus Salinicoccus, with low 16S rRNA gene sequence similarity to the type strains of the five described species of the genus, Salinicoccus alkaliphilus JCM 11311(T) (sequence similarity 96.0 %), Salinicoccus roseus DSM 5351(T) (94.9 %), Salinicoccus jeotgali KCTC 13030(T) (94.8 %), Salinicoccus salsiraiae LMG 22840(T) (94.8 %) and Salinicoccus hispanicus DSM 5352(T) (94.7 %). Together with the phenotypic differences, these results supported the proposal of a novel species of the genus Salinicoccus, Salinicoccus kunmingensis sp. nov., with YIM Y15(T) (=DSM 17847(T) =CGMCC 1.6302(T)) as the type strain. PMID- 17911307 TI - Rubritalea spongiae sp. nov. and Rubritalea tangerina sp. nov., two carotenoid- and squalene-producing marine bacteria of the family Verrucomicrobiaceae within the phylum 'Verrucomicrobia', isolated from marine animals. AB - Two Gram-negative, non-motile, coccoid or rod-shaped, chemoheterotrophic bacteria designated strains YM21-132(T) and YM27-005(T) were isolated from marine animals, and were subjected to a polyphasic taxonomic examination. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the two isolates belong to the genus Rubritalea of the phylum 'Verrucomicrobia' (subdivision 1). The novel isolates shared approximately 97-98 % sequence similarity with each other and showed 93-97 % similarity with Rubritalea species of the family Verrucomicrobiaceae. The level of DNA-DNA relatedness between strains YM21-132(T) and YM27-005(T) was less than 70 %, which is accepted as the phylogenetic definition of a species. Both strains produced reddish carotenoid pigments and squalene. The cell wall peptidoglycan of both strains contained muramic acid and meso-diaminopimelic acid. The G+C contents of the genomic DNA were 48.0 mol% (strain YM21-132(T)) and 50.3 mol% (strain YM27-005(T)). The presence of MK-8 and MK-9 as the major isoprenoid quinones, and iso-C(14 : 0), iso-C(16 : 0) and C(16 : 1)omega7c as the major cellular fatty acids supported the identification of the two novel strains as members of the genus Rubritalea. On the basis of polyphasic taxonomic studies, it was concluded that these strains should be classified as representing two novel, separate species in the genus Rubritalea within the phylum 'Verrucomicrobia', for which the names Rubritalea spongiae sp. nov. (type strain YM21-132(T)=MBIC08281(T)=KCTC 12906(T)) and Rubritalea tangerina sp. nov. (type strain YM27-005(T)=MBIC08282(T)=KCTC 12907(T)) are proposed. PMID- 17911308 TI - Chryseobacterium haifense sp. nov., a psychrotolerant bacterium isolated from raw milk. AB - Strain H38(T) was isolated during a study on the diversity of culturable psychrotolerant bacteria in raw milk. Comparative 16S rRNA gene sequence analysis showed that strain H38(T) belongs to the genus Chryseobacterium (family Flavobacteriaceae, phylum Bacteroidetes), with the highest similarity (96.7 %) to Chryseobacterium hispanicum VP48(T). Strain H38(T) grows aerobically, at 4-41 degrees C, with 0-2.5 % NaCl and at pH 6.5-10.5. Light induces the formation of yellow carotenoid pigments. The dominant cellular fatty acids are 15 : 0 iso (41.6 %), 15 : 0 anteiso (16.6 %) and 17 : 0 iso 3-OH (10.3 %). The G+C content of the DNA is 37.8 mol%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain H38(T) (=LMG 24029(T) =DSM 19056(T)) is classified in the genus Chryseobacterium as the type strain of a novel species, for which the name Chryseobacterium haifense sp. nov. is proposed. PMID- 17911310 TI - Microbacterium sediminicola sp. nov. and Microbacterium marinilacus sp. nov., isolated from marine environments. AB - Two novel Microbacterium species are described on the basis of phenotypic, chemotaxonomic and genotypic studies. The two strains, designated YM10-847(T) and YM11-607(T), were isolated from river sediment and unidentified hydroid, respectively, of a marine lake. The strains were Gram-positive, catalase-positive bacteria with l-ornithine as the diagnostic diamino acid of the peptidoglycan. The acyl type of the peptidoglycan was N-glycolyl. The major menaquinones were MK 10 and MK-11 for YM10-847(T), and MK-11 and MK-12 for YM11-607(T). Mycolic acids were not detected. The DNA G+C content of strains YM10-847(T) and YM11-607(T) was 67.8 and 71.6 mol%, respectively. Comparative 16S rRNA gene sequence analysis revealed that the two strains belong to the genus Microbacterium. DNA-DNA relatedness data showed that YM10-847(T) and YM11-607(T) are two novel species of this genus. On the basis of these results, strains YM10-847(T) and YM11-607(T) represent two novel species of the genus Microbacterium, for which the names Microbacterium sediminicola sp. nov. and Microbacterium marinilacus sp. nov. are proposed. The type strains are YM10-847(T) (=MBIC08264(T)=DSM 18905(T)) and YM11 607(T) (=MBIC07778(T)=DSM 18904(T)), respectively. PMID- 17911309 TI - Mucilaginibacter paludis gen. nov., sp. nov. and Mucilaginibacter gracilis sp. nov., pectin-, xylan- and laminarin-degrading members of the family Sphingobacteriaceae from acidic Sphagnum peat bog. AB - Two facultatively aerobic, heterotrophic bacteria capable of degrading pectin, xylan, laminarin and some other polysaccharides were obtained from the acidic Sphagnum peat bog Bakchar, in western Siberia, Russia, and were designated strains TPT18(T) and TPT56(T). Cells of these isolates are Gram-negative, non motile, long rods that are covered by large capsules. On ageing, they transform into spherical L-forms. Strains TPT18(T) and TPT56(T) are acido- and psychrotolerant organisms capable of growth at pH 4.2-8.2 (with an optimum at pH 6.0-6.5) and at 2-33 degrees C (with an optimum at 20 degrees C). The major fatty acids are iso-C(15 : 0), anteiso-C(15 : 0), iso-C(17 : 0) 3-OH and summed feature 3 (iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c); the quinones are MK-7 and MK-6. Comparative 16S rRNA gene sequence analysis revealed that the novel strains share 97 % sequence similarity and belong to the family Sphingobacteriaceae; however, they are related only distantly to members of the genera Pedobacter (91.8-93.3 % similarity) and Sphingobacterium (89.6-91.2 % similarity). The DNA G+C content of strains TPT18(T) and TPT56(T) is 42.4 and 46.1 mol%, respectively. The low DNA DNA hybridization value (42 %) and a number of phenotypic differences between strains TPT18(T) and TPT56(T) indicated that they represent two separate species. Since the two isolates are clearly distinct from all currently described members of the family Sphingobacteriaceae, we propose a novel genus, Mucilaginibacter gen. nov., containing two novel species, Mucilaginibacter gracilis sp. nov. and Mucilaginibacter paludis sp. nov. The type strains of Mucilaginibacter gracilis and Mucilaginibacter paludis are respectively TPT18(T) (=ATCC BAA-1391(T) =VKM B 2447(T)) and TPT56(T) (=ATCC BAA-1394(T) =VKM B-2446(T)). PMID- 17911311 TI - Desulfothermus okinawensis sp. nov., a thermophilic and heterotrophic sulfate reducing bacterium isolated from a deep-sea hydrothermal field. AB - A novel thermophilic and heterotrophic sulfate-reducing bacterium, strain TFISO9(T), was isolated from a deep-sea hydrothermal field at the Yonaguni Knoll IV in the Southern Okinawa Trough. The cells were motile rods 2.5-5.0 microm in length and 0.6-0.9 microm in width. Strain TFISO9(T) was an obligate heterotroph and reduced sulfate. It grew between 35 and 60 degrees C (optimum 50 degrees C), at pH 5.4-7.9 (optimum pH 5.9-6.4) and with 1.5-4.5 % NaCl (optimum 2.5 %). The fatty acid composition was C(16 : 0) (61.5 %) and 12Me(16 : 0) (38.5 %). The DNA G+C content was 34.9 mol%. The 16S rRNA gene sequence analysis indicated that strain TFISO9(T) belonged to the genus Desulfothermus. Based on physiological and phylogenetic characteristics, strain TFISO9(T) represents a novel species for which the name Desulfothermus okinawensis sp. nov. is proposed. The type strain is TFISO9(T) (=JCM 13304(T)=DSM 17375(T)). PMID- 17911313 TI - Reinekea blandensis sp. nov., a marine, genome-sequenced gammaproteobacterium. AB - A novel heterotrophic, moderately halophilic, strictly aerobic, motile bacterium was isolated from a seawater sample collected at the Blanes Bay Microbial Observatory in the north-western Mediterranean Sea. Analysis of its 16S rRNA gene sequence, retrieved from the whole-genome sequence, showed that this bacterium was most closely related to the single-species genera Reinekea and Saccharospirillum (95 and 94 % sequence similarity, respectively) within the class Gammaproteobacteria. The data from phenotypic, genotypic, chemotaxonomic and phylogenetic analyses supported the creation of a novel species of the genus Reinekea to accommodate this bacterium, for which the name Reinekea blandensis sp. nov. is proposed. The type strain is MED297(T) (=CECT 7120(T) =CCUG 52066(T)). PMID- 17911312 TI - Microbulbifer celer sp. nov., isolated from a marine solar saltern of the Yellow Sea in Korea. AB - A Gram-negative, non-motile, rod-shaped, Microbulbifer-like bacterial strain, ISL 39(T), was isolated from a marine solar saltern of the Yellow Sea in Korea and was subjected to a polyphasic taxonomic investigation. Strain ISL-39(T) grew optimally at pH 7.0-8.0 and 37 degrees C. It contained Q-8 as the predominant ubiquinone and iso-C(15 : 0), C(16 : 0) and iso-C(17 : 0) as the major fatty acids. The DNA G+C content was 57.7 mol%. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain ISL-39(T) belonged to the genus Microbulbifer. Strain ISL-39(T) exhibited 16S rRNA gene sequence similarity values of 94.7-97.5 % with respect to the type strains of four recognized Microbulbifer species. DNA-DNA relatedness data and the differential phenotypic properties and phylogenetic distinctiveness of ISL-39(T) make this strain distinguishable from the recognized Microbulbifer species. On the basis of the phenotypic, phylogenetic and genetic data, strain ISL-39(T) represents a novel species of the genus Microbulbifer, for which the name Microbulbifer celer sp. nov. is proposed. The type strain is ISL-39(T) (=KCTC 12973(T)=CCUG 54356(T)). PMID- 17911314 TI - Rheinheimera texasensis sp. nov., a halointolerant freshwater oligotroph. AB - A Gram-negative, rod-shaped, motile, non-spore-forming bacterium, designated strain A62-14B(T), was isolated from a constant-temperature, spring-fed, freshwater lake. On the basis of the complete 16S rRNA gene sequence, strain A62 14B(T) was shown to belong to the class Gammaproteobacteria, being most closely related to Rheinheimera sp. HTB082 (96.2 % sequence similarity), Rheinheimera baltica (95.01 %), Rheinheimera pacifica (96.35 %), Rheinheimera perlucida and Alishewanella fetalis (95.9 %). The major fatty acids (C(16 : 1)omega7c, 38.56 %; C(16 : 0), 19.04 %; C(12 : 0) 3-OH, 12.83 %; C(18 : 1)omega7c, 7.70 %) and the motility of strain A62-14B(T) support its affiliation to the genus Rheinheimera. The salt intolerance of strain A62-14B(T), together with the results of other physiological and biochemical tests, allowed the differentiation of this strain from the three species of the genus Rheinheimera with validly published names. Therefore strain A62-14B(T) represents a novel species of the genus Rheinheimera, for which the name Rheinheimera texasensis sp. nov. is proposed. The type strain is A62-14B(T) (=ATCC BAA-1235(T)=DSM 17496(T)). The description of the genus Rheinheimera is emended to reflect the halointolerance and freshwater origin of strain A62-14B(T). PMID- 17911315 TI - Salsuginibacillus kocurii gen. nov., sp. nov., a moderately halophilic bacterium from soda-lake sediment. AB - A Gram-positive, endospore-forming, alkali-tolerant, moderately halophilic bacterium, designated strain CH9d(T), was isolated from the sediment of Lake Chagannor in the Inner Mongolia Autonomous Region, China. The cells were rod shaped and motile. Isolate CH9d(T) grew at pH 5.8-10.0 (optimally at pH 8.5), at salinities of 3-20 % (w/v) marine salts (optimally at 10.0 %, w/v) and between 20 and 50 degrees C (optimally at 37 degrees C). The cell wall contained meso diaminopimelic acid and the major respiratory isoprenoid quinone was MK-7. The predominant cellular fatty acids of strain CH9d(T) were anteiso-C(15 : 0), anteiso-C(17 : 0), iso-C(17 : 0) and iso-C(15 : 0) and its polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and two unknown phospholipids. The G+C content of the DNA was 44.7 mol%. Strain CH9d(T) exhibited a 16S rRNA gene sequence similarity value of only 91 % with respect to Thalassobacillus devorans DSM 16966(T) and showed values below 91 % with respect to members of the genera Bacillus, Halobacillus and Marinococcus. Strain CH9d(T) could be clearly differentiated from its closest phylogenetic neighbours on the basis of several phenotypic, genotypic and chemotaxonomic features. Therefore, data from the polyphasic study support the placement of strain CH9d(T) in a novel genus and species, for which the name Salsuginibacillus kocurii gen. nov., sp. nov. is proposed. The type strain is CH9d(T) (=CCM 7365(T)=CECT 7154(T)=CGMCC 1.6287(T)=DSM 18087(T)). PMID- 17911316 TI - Phylogeny and evolution of the family Ectothiorhodospiraceae based on comparison of 16S rRNA, cbbL and nifH gene sequences. AB - The occurrence of genes encoding nitrogenase and ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) was investigated in the members of the family Ectothiorhodospiraceae. This family forms a separate phylogenetic lineage within the Gammaproteobacteria according to 16S rRNA gene sequence analysis and mostly includes photo- and chemoautotrophic halophilic and haloalkaliphilic bacteria. The cbbL gene encoding the large subunit of 'green-like' form I RubisCO was found in all strains, except the type strains of Alkalispirillum mobile and Arhodomonas aquaeolei. The nifH gene encoding nitrogenase reductase was present in all investigated species of the phototrophic genera Ectothiorhodospira, Halorhodospira and Thiorhodospira, but not of the genus Ectothiorhodosinus. Unexpectedly, nifH fragments were also obtained for the chemotrophic species Thioalkalispira microaerophila and Alkalilimnicola halodurans, for which diazotrophic potential has not previously been assumed. The cbbL-, nifH- and 16S rRNA gene-based trees were not highly congruent in their branching patterns since, in the 'RubisCO' and 'nitrogenase' trees, representatives of the Ectothiorhodospiraceae are divided in a number of broadly distributed clusters and branches. However, the data obtained may be regarded as evidence of the monophyletic origin of the cbbL and nifH genes in most species within the family Ectothiorhodospiraceae and mainly corresponded to the current taxonomic structure of this family. The cbbL phylogeny of the chemolithoautotrophic sulfur-oxidizers Thioalkalivibrio nitratireducens and Thioalkalivibrio paradoxus and the nitrifier Nitrococcus mobilis deviated significantly from the 16S-rRNA gene-based phylogeny. These species clustered with one of the duplicated cbbL genes of the purple sulfur bacterium Allochromatium vinosum, a member of the family Chromatiaceae. PMID- 17911317 TI - Anderseniella baltica gen. nov., sp. nov., a novel marine bacterium of the Alphaproteobacteria isolated from sediment in the central Baltic Sea. AB - A bacterial isolate from the Baltic Sea, designated strain BA141(T), was characterized for its physiological and biochemical features, fatty acid profile, pigment spectrum, DNA G+C content and phylogenetic position based on comparative 16S rRNA gene sequence analysis. The strain was isolated from the surface of sediment in a deep basin of the central Baltic Sea. Phylogenetic analysis of the 16S rRNA gene sequence revealed a clear affiliation with the Alphaproteobacteria, and showed that the closest phylogenetic relationship was with the genus Rhodobium. The G+C content of the DNA was 61.2 mol%. Cells of strain BA141(T) were red-pigmented, Gram-negative, pleomorphic to rod-shaped, non-motile and catalase- and oxidase-positive. Growth was observed at salinities of 0.8-6 %, with optimum growth at 1.5-3 %. The temperature range for growth was 10-37 degrees C, with optimum growth at 25-30 degrees C. The fatty acids were dominated by unsaturated fatty acids (>86 %); the number of fatty acids detected was very low, with 18 : 1omega7c (73 %) as the predominant fatty acid; other major fatty acids were 19 : 0 cyclo 8c (10 %) and 16 : 0 (8 %). The pigment spectrum indicated the presence of carotenoids and unknown pigment(s) with an absorption maximum at 430 nm, but not bacteriochlorophyll a. According to phylogenetic analysis based on the 16S rRNA gene sequence and the phenotypic features, strain BA141(T) represents a new genus and species. The name Anderseniella baltica gen. nov., sp. nov. is proposed, with strain BA141(T) (=CIP 109499(T)=LMG 24028(T)) as the type strain. PMID- 17911318 TI - Reclassification of Marinococcus albus Hao et al. 1985 as Salimicrobium album gen. nov., comb. nov. and Bacillus halophilus Ventosa et al. 1990 as Salimicrobium halophilum comb. nov., and description of Salimicrobium luteum sp. nov. AB - A Gram-positive, non-motile, coccoid-shaped, non-spore-forming halophilic bacterial strain, BY-5(T), was isolated from a marine solar saltern in Korea and its taxonomic position was investigated by using a polyphasic approach. The novel strain grew optimally at 37 degrees C and in the presence of 10 % (w/v) NaCl. Strain BY-5(T) had meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, MK-7 as the predominant menaquinone and anteiso-C(15 : 0), iso-C(15 : 0), anteiso-C(17 : 0) and iso-C(17 : 0) as the major fatty acids. The DNA G+C content was 47.9 mol%. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain BY-5(T) formed a coherent cluster with Bacillus halophilus and Marinococcus albus. Strain BY-5(T) exhibited 16S rRNA gene sequence similarity values of 98.7 and 97.4 % to the type strains of B. halophilus and M. albus, respectively. Strain BY-5(T) was distinguished from B. halophilus and M. albus by several phenotypic properties and DNA-DNA relatedness data. On the basis of the combined chemotaxonomic and phylogenetic data, it is proposed that M. albus, B. halophilus and strain BY-5(T) should be placed in a new genus as three separate species. Marinococcus albus and Bacillus halophilus are reclassified in a new genus, Salimicrobium gen. nov., as Salimicrobium album comb. nov. and Salimicrobium halophilum comb. nov., respectively. The type species of the new genus is Salimicrobium album. Strain BY-5(T) (=KCTC 3989(T)=CIP 108918(T)) is placed in the genus Salimicrobium as a novel species Salimicrobium luteum sp. nov. PMID- 17911319 TI - Barcoding ciliates: a comprehensive study of 75 isolates of the genus Tetrahymena. AB - The mitochondrial cytochrome-c oxidase subunit 1 (cox1) gene has been proposed as a DNA barcode to identify animal species. To test the applicability of the cox1 gene in identifying ciliates, 75 isolates of the genus Tetrahymena and three non Tetrahymena ciliates that are close relatives of Tetrahymena, Colpidium campylum, Colpidium colpoda and Glaucoma chattoni, were selected. All tetrahymenines of unproblematic species could be identified to the species level using 689 bp of the cox1 sequence, with about 11 % interspecific sequence divergence. Intraspecific isolates of Tetrahymena borealis, Tetrahymena lwoffi, Tetrahymena patula and Tetrahymena thermophila could be identified by their cox1 sequences, showing <0.65 % intraspecific sequence divergence. In addition, isolates of these species were clustered together on a cox1 neighbour-joining (NJ) tree. However, strains identified as Tetrahymena pyriformis and Tetrahymena tropicalis showed high intraspecific sequence divergence values of 5.01 and 9.07 %, respectively, and did not cluster together on a cox1 NJ tree. This may indicate the presence of cryptic species. The mean interspecific sequence divergence of Tetrahymena was about 11 times greater than the mean intraspecific sequence divergence, and this increased to 58 times when all isolates of species with high intraspecific sequence divergence were excluded. This result is similar to DNA barcoding studies on animals, indicating that congeneric sequence divergences are an order of magnitude greater than conspecific sequence divergences. Our analysis also demonstrated low sequence divergences of <1.0 % between some isolates of T. pyriformis and Tetrahymena setosa on the one hand and some isolates of Tetrahymena furgasoni and T. lwoffi on the other, suggesting that the latter species in each pair is a junior synonym of the former. Overall, our study demonstrates the feasibility of using the mitochondrial cox1 gene as a taxonomic marker for 'barcoding' and identifying Tetrahymena species and some other ciliated protists. PMID- 17911320 TI - Polyphasic re-examination of Yarrowia lipolytica strains and the description of three novel Candida species: Candida oslonensis sp. nov., Candida alimentaria sp. nov. and Candida hollandica sp. nov. AB - The type strain of Yarrowia lipolytica and 38 strains identified as Yarrowia lipolytica, four strains of Candida deformans, including the type and two subcultures of the type, two strains of Candida galli and six unidentified strains that resembled Y. lipolytica were examined by PCR fingerprints using primers M13 and (GAC)5. The same strains, together with four strains of the recently introduced Candida yakushimensis nom. inval., were sequenced for the D1/D2 domain of the 26S rRNA gene and parts of the ITS domain and also studied for their physiological properties. Of the strains identified previously as Y. lipolytica, CBS 2076 had the same fingerprint as the type of C. deformans and strain CBS 4855 was distinct from all other strains. The six strains that resembled Y. lipolytica were separated into two groups distinct from any of the other clades. A total of six groups obtained by fingerprint and sequence data were evaluated by performing DNA reassociation reactions. Mating experiments among the 35 strains of Y. lipolytica sensu stricto showed that 15 strains represented one mating type and 16 strains represented the opposite mating type, while four strains were self-sporulating. Teleomorph states were not produced by C. deformans, C. galli or any of the unidentified isolates. However, positive mating reactions were rarely observed in crosses among C. galli and some strains of Y. lipolytica and C. deformans. Consequently, sharing the same mating type system, C. deformans and C. galli could be considered anamorphs of unnamed Yarrowia species. Results from PCR fingerprints, sequencing and mating studies support the grouping of the studied strains into Y. lipolytica, C. galli, C. deformans, C. yakushimensis nom. inval. and three novel species in the Yarrowia clade: Candida oslonensis sp. nov. (type strain CBS 10146(T) =NRRL Y-48252(T); Mycobank number MB 510769), Candida alimentaria sp. nov. (type strain CBS 10151(T) =NRRL Y-48253(T); Mycobank number MB 510770) and Candida hollandica sp. nov. (type strain CBS 4855(T) =NRRL Y-48254(T); Mycobank number MB 510771). PMID- 17911321 TI - Recommended minimal standards for describing new taxa of the family Halomonadaceae. AB - Following Recommendation 30b of the Bacteriological Code (1990 Revision), a proposal of minimal standards for describing new taxa within the family Halomonadaceae is presented. An effort has been made to evaluate as many different approaches as possible, not only the most conventional ones, to ensure that a rich polyphasic characterization is given. Comments are given on the advantages of each particular technique. The minimal standards are considered as guidelines for authors to prepare descriptions of novel taxa. The proposals presented here have been endorsed by the International Committee on Systematics of Prokaryotes Subcommittee on the Taxonomy of Halomonadaceae. PMID- 17911322 TI - Recommendation for the conservation of the name Streptomyces scabies. Request for an Opinion. AB - The primary streptomycete inciting common scab of potato was first legitimately described by Thaxter in 1892 as 'Oospora scabies', preserving the spelling of an epithet in use since 1846. The name Streptomyces scabies, dating to 1948, was revived in 1989, but changed to Streptomyces scabiei in 1997 to follow grammatical convention. Considering the long-established use and general recognition of 'scabies', it is proposed that the original epithet be conserved. PMID- 17911323 TI - Data-based articles and duplicate publication. PMID- 17911325 TI - Envisioning nursing in 2050 through the eyes of nurse theorists: Rosemarie Rizzo Parse and Martha E. Rogers. AB - Rosemarie Rizzo Parse and Rogerian scholar, Violet Malinski, on behalf of Martha E. Rogers, envision how their respective theoretical ideas will be expressed in 2050. Parse introduces several changes highlighting the idea of indivisible cocreation in the ontology of the humanbecoming school of thought and concomitant changes in the wording of its principles. Notions of human freedom and dignity will be extremely important given increasingly mechanized healthcare. Malinski describes Rogerian formulations for practice and research that will continue to facilitate well-being for humankind and the environment. She also addresses challenges and opportunities for an evolving science of unitary human beings. PMID- 17911326 TI - The humanbecoming school of thought in 2050. PMID- 17911327 TI - The subjective-objective rhythm of research. AB - Cowling and Shattell share their thoughts on participatory inquiry within a worldview that presents reality as a co-creative process that recognizes a multiplicity of ways of knowing. They present the five dimensions of a participatory worldview as it informs the emergence of story in the research process. They conclude their column with examples from their own work, Cowling's on women survivors of child abuse, Shattell's on Latino mental health. PMID- 17911328 TI - The ethics of nurse migration: an evolution of community change. AB - International nursing migration, or the movement of people across international boundaries, has enormous implications for the discipline of nursing. This column focuses on ethical assumptions and possible implications for community change as individual or group. Discussion follows with the illumination of community change paradoxes, implications for policy development, nursing education, and nursing practice from the nursing theoretical perspective of the humanbecoming school of thought. PMID- 17911329 TI - Using case studies based on a nursing conceptual model to teach medical-surgical nursing. AB - Nurse educators are continually challenged to develop teaching strategies that enhance students' critical thinking, problem-solving, and decision-making skills. Case studies are a creative learning strategy that fosters these skills through the use of in-depth descriptions of realistic clinical situations. Conceptual models of nursing provide a unique body of knowledge that can be used to guide construction of case studies and enhance application of didactic course content to nursing practice. In this column, the author discusses the use of case studies constructed within the context of the Roy adaptation model for a senior level medical-surgical nursing course. PMID- 17911330 TI - Practice in 2050: caring science and a synthesis of views. AB - In this column nurses influenced by Watson's human caring theory and Eriksson's theory of caritative caring share their ideas about what nursing theory-guided practice will be like in 2050. Following these contributions about caring theories in 2050, is a synthesis of threads of relatedness across the volume 20 Practice Applications columns-all of which focus on possibilities for nursing theory-guided practice in 2050. PMID- 17911331 TI - Nursing, healthcare, and culture: a view of the year 2050 from Botswana and Israel. AB - This column presents a dialogue with nurse scholars Motshedisi B. Sabone from Botswana and Freda DeKeyser Ganz from Israel, who share their respective visions and understanding of each country's vision of nursing, healthcare, and quality of life in the year 2050. PMID- 17911332 TI - Recovering from abuse: a comparison of three paths. AB - Nursing, and other health professions, have for many years been generally well educated to be on the alert for, identify, and report abuse, particularly of children. What many nurses are less prepared for, and less comfortable with, is how to help adult patients who were victims of abuse in the past, and who still suffer from its consequences. The purpose of this article is to compare and contrast three different pathways to recovery for persons living with the legacy of abuse and victimization. The three paths are the traditional psychiatric model, spiritually-based self-help, and the humanbecoming theory. PMID- 17911333 TI - Review of Roy adaptation model-based qualitative research. AB - The aim of this paper is to examine the role of qualitative research in the development of the Roy adaptation model. An exploration of the findings from qualitative research using Roy's adaptation model from 1995 to 2005 is compared with the findings and recommendations from a previous review over a 25-year period (1970-1995). The usefulness of qualitative methods in furthering nursing theory is highlighted. Findings from both reviews support the assumptions of the model while generating new information and demonstrating the valuable place of qualitative research in promoting nursing science. PMID- 17911334 TI - Ericksonian hypnosis in chronic care support groups: a Rogerian exploration of power and self-defined health-promoting goals. AB - This Rogerian study examined how traditional and Ericksonian hypnotherapeutic support groups facilitated self-defined health-promoting goals and power as knowing participation in change for 49 participants with chronic physical illness. The participants were randomly assigned to either a traditional support group or an Ericksonian hypnotherapeutic support group. Measurements of power and self-defined health-promoting goals were obtained seven times over a 10-week period. The results indicated that both the traditional support groups and the Ericksonian hypnotherapeutic support groups experienced significantly enhanced power and progressed significantly toward their health-promoting goals. Correlations for the self-defined health-promoting goals and power progressively and significantly increased through time. This study supports Barrett's claim that power relates to health. PMID- 17911335 TI - Nursing as a sign of hope and contradiction in the land of the children of Abraham. AB - Israel, the Gaza Strip, and the West Bank share a small land area and the birthplace of three of the world's great monotheistic religions: Judaism, Christianity, and Islam. It is a land of paradoxes: ancient and modern, religious and secular. Nursing in this unique geographic and historical place struggles with many of the same issues and challenges facing nursing in other modern countries, but it also shines with dramatic episodes of nursing's ability to be a sign of hope and contradiction. PMID- 17911336 TI - Nursing in Israel. AB - Nurses in Israel struggle with many of the same problems faced by nurses in other parts of the world, such as increased use of technology, overwhelming amounts of information, and demands for high quality of services to larger numbers of people within tighter budgets. In addition to the aging of the general population, the country has welcomed large numbers of immigrants. The nation's expenditures for healthcare and nursing education have, at times, had to take a back seat to the government's efforts to house new immigrants, to relocate groups, and to defend the nation against politically motivated violence and attacks. All of this is in the context of regional conflicts and international debates. PMID- 17911337 TI - Living as an oldest old in Rio de Janeiro: the lived experience told. AB - This phenomenological-hermeneutic study was an inquiry into the lived experience of 16 very old persons who were living in their own homes. The nursing perspective was Gadow's existential advocacy for nursing practice. The study findings led to the essence of the phenomenon: pragmatic transition to old age, within a restricted future. Heideggerian concepts were applied and the findings were discussed in relation to the guiding theoretical perspective and related literature. Understanding the meaning of being a very old person is fundamental to developing nursing interventions to help people remain independent, with a sense of self-determination, for as long as possible. PMID- 17911338 TI - Adrenocortical dysregulation as a major player in insulin resistance and onset of obesity. AB - The aim of this review is to explore the dysregulation of adrenocortical secretions as a major contributor in the development of obesity and insulin resistance. Disturbance of adipose tissue physiology is one of the primary events in the development of pathologies associated with the metabolic syndrome, such as obesity and type 2 diabetes. Several studies indicate that alterations in metabolism of glucocorticoids (GC) and androgens, as well as aldosterone in excess, are involved in the emergence of metabolic syndrome. Cross talk among adipose tissue, the hypothalamo-pituitary complex, and adrenal gland activity plays a major role in the control of food intake, glucose metabolism, lipid storage, and energy balance. Perturbation of this cross talk induces alterations in the regulatory mechanisms of adrenocortical steroid synthesis, secretion, degradation, and/or recycling, at the level of the zonae glomerulosa (aldosterone), fasciculata (GC and GC metabolites), and reticularis (androgens and androgen precursors DHEA and DHEAS). As a whole, these adrenocortical perturbations contribute to the development of metabolic syndrome at both the paracrine and systemic level by favoring the physiological dysregulation of organs responsive to aldosterone, GC, and/or androgens, including adipose tissue. PMID- 17911339 TI - Increased intrahepatic triglyceride is associated with peripheral insulin resistance: in vivo MR imaging and spectroscopy studies. AB - Recent studies have indicated that the mass/content of intramyocellular lipid (IMCL), intrahepatic triglyceride (IHTG), visceral fat (VF), and even deep abdominal subcutaneous fat (SF) may all be correlated with insulin resistance. Since simultaneous measurements of these parameters have not been reported, the relative strength of their associations with insulin action is not known. Therefore, the goals of this study were 1) to simultaneously measure IMCL, IHTG, VF, and abdominal SF in the same nondiabetic individuals using noninvasive (1)H magnetic resonance spectroscopy (MRS) and magnetic resonance imaging (MRI) and 2) to examine how these fat stores are correlated with systemic insulin sensitivity as measured by whole body glucose disposal (R(d)) during euglycemic hyperinsulinemic clamp studies. Positive correlations were observed among IMCL, IHTG, and VF. There were significant inverse correlations between whole body R(d) and both IMCL and VF. Notably, there was a particularly tight inverse correlation between IHTG and whole body R(d) (r = -0.86, P < 0.001), consistent with an association between liver fat and peripheral insulin sensitivity. This novel finding suggests that hepatic triglyceride accumulation has important systemic consequences that may adversely affect insulin sensitivity in other tissues. PMID- 17911340 TI - Common and specific effects of the two major forms of prolactin in the rat testis. AB - Prolactin (PRL) has both stimulatory and inhibitory effects on testicular function, a finding we hypothesized may be related in some part to the form of the hormone present or administered. In the analysis of the pituitary secretion profiles of early pubescent vs. mature male rats, we found PRL released from early pubescent pituitaries had about twice the degree of phosphorylation. Treatment of mature males with either unmodified PRL (U-PRL) or phosphorylated PRL (via the molecular mimic S179D PRL) for a period of 4 wk (circulating level of approximately 50 ng/ml) showed serum testosterone decreased by approximately 35% only by treatment with the phospho-mimic S179D PRL. Given the specificity of this effect, it was initially surprising that both forms of PRL decreased testicular expression of 3beta-hydroxysteroid dehydrogenase and steroidogenic acute regulatory protein. Both forms also increased expression of the luteinizing hormone receptor, but only S179D PRL increased the ratio of short to long PRL receptors. Endogenous PRL and luteinizing hormone levels were unchanged in all groups in this time frame, suggesting that effects on steroidogenic gene expression were directly on the testis. Terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end-labeling analysis combined with staining for 3beta-hydroxysteroid dehydrogenase and morphometric analysis showed that S179D PRL, but not U-PRL, increased apoptosis of Leydig cells, a finding supported by increased staining for Fas and Fas ligand in the testicular interstitium, providing an explanation for the specific effect on testosterone. S179D PRL, but not U-PRL, also increased apoptosis of primary spermatogonia, and U-PRL, but not S179D PRL, decreased apoptosis of elongating spermatids. Thus, in mature males, hyperprolactinemic levels of both forms of PRL have common effects on steroidogenic proteins, but specific effects on the apoptosis of Leydig and germ cells. PMID- 17911341 TI - Skeletal muscle capillary responses to insulin are abnormal in late-stage diabetes and are restored by angiotensin-converting enzyme inhibition. AB - Acute physiological hyperinsulinemia increases skeletal muscle capillary blood volume (CBV), presumably to augment glucose and insulin delivery. We hypothesized that insulin-mediated changes in CBV are impaired in type 2 diabetes mellitus (DM) and are improved by angiotensin-converting enzyme inhibition (ACE-I). Zucker obese diabetic rats (ZDF, n = 18) and control rats (n = 9) were studied at 20 wk of age. One-half of the ZDF rats were treated with quinapril (ZDF-Q) for 15 wk prior to study. CBV and capillary flow in hindlimb skeletal muscle were measured by contrast-enhanced ultrasound (CEU) at baseline and at 30 and 120 min after initiation of a euglycemic hyperinsulinemic clamp (3 mU.min(-1).kg(-1)). At baseline, ZDF and ZDF-Q rats were hyperglycemic and hyperinsulinemic vs. controls. Glucose utilization in ZDF rats was 60-70% lower (P < 0.05) than in controls after 30 and 120 min of hyperinsulinemia. In ZDF-Q rats, glucose utilization was impaired at 30 min but similar to controls at 120 min. Basal CBV was lower in ZDF and ZDF-Q rats compared with controls (13 +/- 4, 7 +/- 3, and 9 +/- 2 U, respectively). With hyperinsulinemia, CBV increased by about twofold in control animals at 30 and 120 min, did not change in ZDF animals, and increased in ZDF-Q animals only at 120 min to a level similar to controls. Anatomic capillary density on immunohistology was not different between groups. We conclude that insulin-mediated capillary recruitment in skeletal muscle, which participates in glucose utilization, is impaired in animals with DM and can be partially reversed by chronic ACE-I therapy. PMID- 17911343 TI - Induction of endoplasmic reticulum stress-induced beta-cell apoptosis and accumulation of polyubiquitinated proteins by human islet amyloid polypeptide. AB - The islet in type 2 diabetes is characterized by an approximately 60% beta-cell deficit, increased beta-cell apoptosis, and islet amyloid derived from islet amyloid polypeptide (IAPP). Human IAPP (hIAPP) but not rodent IAPP (rIAPP) forms toxic oligomers and amyloid fibrils in an aqueous environment. We previously reported that overexpression of hIAPP in transgenic rats triggered endoplasmic reticulum (ER) stress-induced apoptosis in beta-cells. In the present study, we sought to establish whether the cytotoxic effects of hIAPP depend on its propensity to oligomerize, rather than as a consequence of protein overexpression. To accomplish this, we established a novel homozygous mouse model overexpressing rIAPP at a comparable expression rate and, on the same background, as a homozygous transgenic hIAPP mouse model previously reported to develop diabetes associated with beta-cell loss. We report that by 10 wk of age hIAPP mice develop diabetes with a deficit in beta-cell mass due to increased beta-cell apoptosis. The rIAPP transgenic mice counterparts do not develop diabetes or have decreased beta-cell mass. Both rIAPP and hIAPP transgenic mice have increased expression of BiP, but only hIAPP transgenic mice have elevated ER stress markers (X-box-binding protein-1, nuclear localized CCAAT/enhancer binding-protein homologous protein, active caspase-12, and accumulation of ubiquitinated proteins). These findings indicate that the beta-cell toxic effects of hIAPP depend on the propensity of IAPP to aggregate, but not on the consequence of protein overexpression. PMID- 17911342 TI - Role of nitrosative stress in early neuropathy and vascular dysfunction in streptozotocin-diabetic rats. AB - Evidence for important roles of the highly reactive oxidant peroxynitrite in diabetic complications is emerging. We evaluated the role of peroxynitrite in early peripheral neuropathy and vascular dysfunction in STZ-diabetic rats. In the first dose-finding study, control and STZ-diabetic rats were maintained with or without the potent peroxynitrite decomposition catalyst Fe(III)tetrakis-2-(N triethylene glycol monomethyl ether) pyridyl porphyrin (FP15) at 3, 5, or 10 mg.kg(-1).day(-1) in the drinking water for 4 wk after an initial 2 wk without treatment for assessment of early neuropathy. In the second study with similar experimental design, control and STZ-diabetic rats were maintained with or without FP15, 5 mg.kg(-1).day(-1), for vascular studies. Rats with 6-wk duration of diabetes developed motor and sensory nerve conduction velocity deficits, mechanical hyperalgesia, and tactile allodynia in the absence of small sensory nerve fiber degeneration. They also had increased nitrotyrosine and poly(ADP ribose) immunofluorescence in the sciatic nerve and dorsal root ganglia. All these variables were dose-dependently corrected by FP15, with minimal differences between the 5 and 10 mg.kg(-1).day(-1) doses. FP15, 5 mg.kg(-1).day(-1), also corrected endoneurial nutritive blood flow and nitrotyrosine, but not superoxide, fluorescence in aorta and epineurial arterioles. Diabetes-induced decreases in acetylcholine-mediated relaxation by epineurial arterioles and coronary and mesenteric arteries, as well as bradykinin-induced relaxation by coronary and mesenteric arteries, were alleviated by FP15 treatment. The findings reveal the important role of nitrosative stress in early neuropathy and vasculopathy and provide the rationale for further studies of peroxynitrite decomposition catalysts in long-term diabetic models. PMID- 17911344 TI - Effects of lycopene on the induction of foam cell formation by modified LDL. AB - The effect of lycopene on macrophage foam cell formation induced by modified low density lipoprotein (LDL) was studied. Human monocyte-derived macrophages (HMDM) were incubated with lycopene in the presence or absence of native LDL (nLDL) or LDL modified by oxidation (oxLDL), aggregation (aggLDL), or acetylation (acLDL). The cholesterol content, lipid synthesis, scavenger receptor activity, and the secretion of inflammatory [interleukin (IL)-1beta and tumor necrosis factor (TNF) alpha] and anti-inflammatory (IL-10) cytokines was determined. Lycopene was found to decrease the synthesis of cholesterol ester in incubations without LDL or with oxLDL while triacylglycerol synthesis was reduced in the presence of oxLDL and aggLDL. Scavenger receptor activity as assessed by the uptake of acLDL was decreased by approximately 30% by lycopene. In addition, lycopene inhibited IL-10 secretion by up to 74% regardless of the presence of nLDL or aggLDL but did not affect IL-1beta or TNF-alpha release. Lycopene also reduced the relative abundance of mRNA transcripts for scavenger receptor A (SR-A) in THP-1 macrophages treated with aggLDL. These findings suggest that lycopene may reduce macrophage foam cell formation induced by modified LDL by decreasing lipid synthesis and downregulating the activity and expression of SR-A. However, these effects are accompanied by impaired secretion of the anti-inflammatory cytokine IL-10, suggesting that lycopene may also exert a concomitant proinflammatory effect. PMID- 17911345 TI - Increased TNFalpha and CCAAT/enhancer-binding protein homologous protein with aging predispose preadipocytes to resist adipogenesis. AB - Fat depot sizes peak in middle age but decrease by advanced old age. This phenomenon is associated with ectopic fat deposition, decreased adipocyte size, impaired differentiation of preadipocytes into fat cells, decreased adipogenic transcription factor expression, and increased fat tissue inflammatory cytokine generation. To define the mechanisms contributing to impaired adipogenesis with aging, we examined the release of TNFalpha, which inhibits adipogenesis, and the expression of CCAAT/enhancer-binding protein (C/EBP) homologous protein (CHOP), which blocks activity of adipogenic C/EBP family members, in preadipocytes cultured from young, middle-aged, and old rats. Medium conditioned by fat tissue, as well as preadipocytes, from old rats impeded lipid accumulation by preadipocytes from young animals. More TNFalpha was released by preadipocytes from old than young rats. Differences in TNFalpha-converting enzyme, TNFalpha degradation, or the presence of macrophages in cultures were not responsible. TNFalpha induced rat preadipocyte CHOP expression. CHOP was higher in undifferentiated preadipocytes from old than younger animals. Overexpression of CHOP in young rat preadipocytes inhibited lipid accumulation. TNFalpha short interference RNA reduced CHOP and partially restored lipid accumulation in old rat preadipocytes. CHOP normally increases during late differentiation, potentially modulating the process. This late increase in CHOP was not affected substantially by aging: CHOP was similar in differentiating preadipocytes and fat tissue from old and young animals. Hypoglycemia, which normally causes an adaptive increase in CHOP, was less effective in inducing CHOP in preadipocytes from old than younger animals. Thus increased TNFalpha release by undifferentiated preadipocytes with elevated basal CHOP contributes to impaired adipogenesis with aging. PMID- 17911346 TI - Greater glycogen utilization during 1- than 2-adrenergic receptor stimulation in the isolated perfused rat heart. AB - Differences in energy metabolism during beta(1)- and beta(2)-adrenergic receptor (AR) stimulation have been shown to translate to differences in the elicited functional responses. It has been suggested that differential access to glycogen during beta(1)- compared with beta(2)-AR stimulation may influence the peak functional response and modulation of the response during sustained adrenergic stimulation. Interleaved (13)C- and (31)P-NMR spectroscopy was used during beta(1)- and beta(2)-AR stimulation at matched peak workload (2.5 times baseline) in the isolated perfused rat heart to monitor glycogen levels, phosphorylation potential, and intracellular pH. Simultaneous measurements of left ventricular (LV) function [LV developed pressure (LVDP)], heart rate (HR), and rate-pressure product (RPP = LVDP x HR) were also performed. The heart was perfused under both substrate-free (SF) conditions and with exogenous glucose (G). The greater glycogenolysis was observed during beta(1)- than beta(2)-AR stimulation with G (54% vs. 38% reduction, P = 0.006) and SF (92% vs. 79% reduction, P = 0.04) perfusions. The greater beta(1)-AR-mediated glycogenolysis was correlated with greater ability to sustain the initial contractile response. However, with SF perfusion, the duration of this ability was limited: excessive early glycogen depletion caused an earlier decline in LVDP and phosphorylation potential during beta(1)- than beta(2)-AR stimulation. Therefore, endogenous glycogen stores are depleted earlier and to a greater extent, despite a slightly weaker overall inotropic response, during beta(1)- than beta(2)-AR stimulation. These findings are consistent with beta(1)-AR-specific PKA-dependent glycogen phosphorylase kinase signaling. PMID- 17911347 TI - Primary hyperparathyroidism is associated with increased circulating bone marrow derived progenitor cells. AB - Recently, parathyroid hormone (PTH) was shown to support survival of progenitor cells in bone marrow. The release of progenitor cells occurs in physiological and pathological conditions and was shown to contribute to neovascularization in tumors and ischemic tissues. In the present study we sought to investigate prospectively the effect of primary hyperparathyroidism (PHPT) on mobilization of bone marrow-derived progenitor cells. In 22 patients with PHPT and 10 controls, defined subpopulations of circulating bone marrow-derived progenitor cells (BMCs) were analyzed by flow cytometry (CD45(+)/CD34(+)/CD31(+) cells indicating endothelial progenitor cells, CD45(+)/CD34(+)/c-kit(+) cells indicating hematopoietic stem cells, and CD45(+)/CD34(+)/CXCR4(+) cells indicating progenitor cells with the homing receptor CXCR4). Cytokine serum levels (SCF, SDF 1, VEGF, EPO, and G-CSF) were assessed using ELISA. Levels of PTH and thyroid hormone as well as serum electrolytes, renal and liver parameters, and blood count were analyzed. Our data show for the first time a significant increase of circulating BMCs and an upregulation of SDF-1 and VEGF serum levels in patients with PHPT. The number of circulating BMCs returned to control levels measured 16.7 +/- 2.3 mo after surgery. There was a positive correlation of PTH levels with the number of CD45(+)/CD34(+)/CD31(+), CD45(+)/CD34(+)/c-kit(+), and CD45(+)/CD34(+)/CXCR4(+) cells. However, there was no correlation between cytokine serum concentrations (SDF-1, VEGF) and circulating BMCs. Serum levels of G-CSF, EPO, and SCF known to mobilize BMCs were even decreased or remained unchanged, suggesting a direct effect of PTH on stem cell mobilization. Our data suggest a new function of PTH mobilizing BMCs into peripheral blood. PMID- 17911348 TI - Nonobese, insulin-deficient Ins2Akita mice develop type 2 diabetes phenotypes including insulin resistance and cardiac remodeling. AB - Although insulin resistance has been traditionally associated with type 2 diabetes, recent evidence in humans and animal models indicates that insulin resistance may also develop in type 1 diabetes. A point mutation of insulin 2 gene in Ins2(Akita) mice leads to pancreatic beta-cell apoptosis and hyperglycemia, and these mice are commonly used to investigate type 1 diabetes and complications. Since insulin resistance plays an important role in diabetic complications, we performed hyperinsulinemic-euglycemic clamps in awake Ins2(Akita) and wild-type mice to measure insulin action and glucose metabolism in vivo. Nonobese Ins2(Akita) mice developed insulin resistance, as indicated by an approximately 80% reduction in glucose infusion rate during clamps. Insulin resistance was due to approximately 50% decreases in glucose uptake in skeletal muscle and brown adipose tissue as well as hepatic insulin action. Skeletal muscle insulin resistance was associated with a 40% reduction in total GLUT4 and a threefold increase in PKCepsilon levels in Ins2(Akita) mice. Chronic phloridzin treatment lowered systemic glucose levels and normalized muscle insulin action, GLUT4 and PKCepsilon levels in Ins2(Akita) mice, indicating that hyperglycemia plays a role in insulin resistance. Echocardiography showed significant cardiac remodeling with ventricular hypertrophy that was ameliorated following chronic phloridzin treatment in Ins2(Akita) mice. Overall, we report for the first time that nonobese, insulin-deficient Ins2(Akita) mice develop type 2 diabetes phenotypes including peripheral and hepatic insulin resistance and cardiac remodeling. Our findings provide important insights into the pathogenesis of metabolic abnormalities and complications affecting type 1 diabetes and lean type 2 diabetes subjects. PMID- 17911349 TI - A computer model of gluconeogenesis and lipid metabolism in the perfused liver. AB - A mathematical model of the perfused rat liver was developed to predict intermediate metabolite concentrations and fluxes in response to changes in various substrate concentrations in the perfusion medium. The model simulates gluconeogenesis in the liver perfused separately with lactate and pyruvate and the combination of these substrates with fatty acids (oleate). The model consists of key reactions representing gluconeogenesis, glycolysis, fatty acid metabolism, tricarboxylic acid cycle, oxidative phosphorylation, and ketogenesis. Michaelis Menten-type kinetic expressions, with control by ATP/ADP, are used for many of the reactions. For key regulated reactions (fructose-1,6-bisphosphatase, phosphofructokinase, pyruvate carboxylase, pyruvate dehydrogenase complex, and pyruvate kinase), rate expressions were developed that incorporate allosteric effectors, specific substrate relationships (e.g., cooperative binding), and/or phosphorylation/dephosphorylation using in vitro enzyme activity data and knowledge of the specific mechanisms. The model was independently validated by comparing model predictions with 10 sets of experimental data from 7 different published works, with no parameter adjustments. The simulations predict the same trends, in terms of stimulation of substrate uptake by fatty acid addition, as observed experimentally. In general, the major metabolic indicators calculated by the model are in good agreement with experimental results. For example, the simulated glucose/pyruvate mass yield is 43% compared with the average of 45% reported in the literature. The model accurately predicts the specific time constants of the glucose response (2.5-4 min) and the dynamic behavior of substrate and product fluxes. It is expected that this model will be a useful tool for analyzing the complex relationships between carbohydrate and fat metabolism. PMID- 17911350 TI - Ghrelin inhibits sympathetic nervous activity in sepsis. AB - Our previous studies have shown that norepinephrine (NE) upregulates proinflammatory cytokines by activating alpha(2)-adrenoceptor. Therefore, modulation of the sympathetic nervous system represents a novel treatment for sepsis. We have also shown that a novel stomach-derived peptide, ghrelin, is downregulated in sepsis and that its intravenous administration decreases proinflammatory cytokines and mitigates organ injury. However, it remains unknown whether ghrelin inhibits sympathetic activity through central ghrelin receptors [i.e., growth hormone secretagogue receptor 1a (GHSR-la)] in sepsis. To study this, sepsis was induced in male rats by cecal ligation and puncture (CLP). Ghrelin was administered through intravenous or intracerebroventricular injection 30 min before CLP. Our results showed that intravenous administration of ghrelin significantly reduced the elevated NE and TNF-alpha levels at 2 h after CLP. NE administration partially blocked the inhibitory effect of ghrelin on TNF-alpha in sepsis. GHSR-la inhibition by the administration of a GHSR-la antagonist, [d Arg(1),d-Phe(5), d-Trp(7,9),Leu(11)]substance P, significantly increased both NE and TNF-alpha levels even in normal animals. Markedly elevated circulating levels of NE 2 h after CLP were also significantly decreased by intracerebroventricular administration of ghrelin. Ghrelin's inhibitory effect on NE release was completely blocked by intracerebroventricular injection of the GHSR-1a antagonist or a neuropeptide Y (NPY)/Y(1) receptor antagonist. However, ghrelin's downregulatory effect on TNF-alpha release was only partially diminished by these agents. Thus ghrelin has sympathoinhibitory properties that are mediated by central ghrelin receptors involving a NPY/Y1 receptor-dependent pathway. Ghrelin's inhibitory effect on TNF-alpha production in sepsis is partially because of its modulation of the overstimulated sympathetic nerve activation. PMID- 17911351 TI - Sympathetic nervous system activity may link hyperphagia and fat deposition in human obesity. PMID- 17911353 TI - Renal calcium stones: insights from the control of bone mineralization. AB - Extracellular pyrophosphate (PPi) plays a central role in the control of normal bone mineralization since it antagonizes inorganic phosphate in the promotion of hydroxyapatite deposition. Studies using knock-out mice have established the functional importance of PPi generation via nucleotide pyrophosphatase phosphodiesterases (NPP) and of PPi transmembrane transport by the progressive ankylosis (ANK) protein. Tissue non-specific alkaline phosphatase activity counteracts this by hydrolysis of PPi to inorganic phosphate. The molecular nature and transport function of ANK are reviewed. A close parallel is drawn between the controlled mineralization of bone and the prevention of abnormal calcium crystal deposition within the kidney, especially when concentrated urine is produced. Pyrophosphate is present in urine, and ANK is expressed in the cortical collecting duct where PPi transport to both the tubular lumen and the renal interstitium may occur. Pyrophosphate may also be generated here by nucleoside triphosphate diphosphohydrolases (NTPD2 and 3) together with NPP1. Alkaline phosphatase activity is restricted to the proximal nephron, remote from these sites of PPi generation, transport and function. The physiological importance of PPi generation and transport in preventing idiopathic calcium renal stone disease and nephrocalcinosis now needs to be established. PMID- 17911352 TI - Benefits of lifestyle modification in NAFLD. PMID- 17911354 TI - Heart rate variability in mice: a theoretical and practical guide. AB - The mouse is the animal model principally used to study biological processes in mammals. The mutation, overexpression or knockout of one or several genes can provide insight into human disease. In cardiovascular research, evaluation of autonomic nervous function is an essential tool for a better understanding of the pathophysiological conditions in which cardiomyopathy arises and develops. Analysis of heart rate variability is the least invasive method to evaluate the sympathovagal balance on the sino-atrial level. The need to perform this technique on freely moving mice emerged in the 1990s, but despite previous studies it has been difficult to set up and standardize a common protocol. The multitudes of techniques used, plus subtle differences in methodology, impede the comparison and clear interpretation of results. This article aims to make a survey of heart rate variability analysis and to establish a standardized protocol for the assessment of the autonomic neural regulation of heart rate in mice. PMID- 17911355 TI - Resistance training in men is associated with increased arterial stiffness and blood pressure but does not adversely affect endothelial function as measured by arterial reactivity to the cold pressor test. AB - Resistance training is a popular mode of exercise, but may result in stiffening of the central arteries. Changes in carotid artery diameter were determined using the cold pressor test (CPT), which results in production of nitric oxide via sympathetic activation and is one of the novel methods available for assessing endothelial function in the carotid artery. To investigate the effect of resistance training on endothelial function, we designed a cross-sectional study of carotid arterial vasoreactivity to CPT in men participating in regular resistance training with increased carotid arterial stiffness compared with age matched control subjects. Twelve resistance-trained middle-aged men (age 38.7 +/- 1.7 years) and 17 age-matched control subjects (age 36.8 +/- 1.2 years) were studied. The direction and magnitude of changes in carotid artery diameter were measured by B-mode ultrasonography during sympathetic stress induced by submersion of the foot in ice slush for 90 s. Carotid arterial beta-stiffness index, and systolic and mean arterial blood pressure were higher (7.7 +/- 0.7 versus 6.0 +/- 0.4 arbitrary units, 116 +/- 2 versus 131 +/- 4 mmHg and 86 +/- 2 versus 95 +/- 2 mmHg, respectively, all P < 0.05) in the resistance training group compared with control subjects. There were, however, no significant differences in the amount or percentage change in carotid artery diameter in CPT between the two groups (resistance training group, 0.33 +/- 0.07 mm and 5.2 +/- 1.1%; control group, 0.37 +/- 0.06 mm and 5.8 +/- 0.9%, respectively). These findings suggest that while carotid arterial stiffening and higher blood pressure are observed in regular resistance-trained men, these are not associated with abnormalities in carotid arterial vasoreactivity to sympathetic stimulus, which implies intact endothelial function. PMID- 17911356 TI - Effects of hydralazine on the pulmonary vasculature and respiratory control in humans. AB - This study sought: (1) to clarify the effects of hydralazine on both the pulmonary vasculature and respiratory control in euoxia and hypoxia in healthy humans; and (2) to determine whether hydralazine alters the expression of genes regulated by hypoxia-inducible factor 1 (HIF-1). Ten volunteers participated in two 2 day protocols. Hydralazine (25 mg) or placebo was administered at 1 pm and 11 pm on the first day, and at 1 pm on the second day. In the mornings and afternoons of both days, we measured plasma vascular endothelial growth factor (VEGF) and erythropoietin (EPO) concentrations (both HIF-1-regulated gene products), systemic arterial blood pressure, and changes in heart rate, cardiac output, maximal systolic pressure difference across the tricuspid valve (delta Pmax) and ventilation in response to 20 min of isocapnic hypoxia. Recent hydralazine: (1) decreased diastolic blood pressure; (2) increased heart rate and cardiac output in euoxia and hypoxia whilst having no effect on delta Pmax; and (3) increased the ventilatory sensitivity to hypoxia. Hydralazine had no effect on plasma EPO or VEGF concentration. We conclude that hydralazine increases the sensitivity of the ventilatory response to hypoxia, but lacks any effect on the pulmonary vasculature at the dose studied. It did not affect the expression of HIF-1-regulated genes. PMID- 17911357 TI - Experimental hyperthyroidism increases expression of parathyroid hormone-related peptide and type-1 parathyroid hormone receptor in rat ventricular myocardium of the Langendorff ischaemia-reperfusion model. AB - Parathyroid hormone-related peptide (PTHrP) is released under ischaemic conditions and it improves contractile function of stunned myocardium. The actions of PTHrP are mediated primarily by the type 1 parathyroid hormone receptor (PTH.1R), while PTHrP and PTH.1R expression levels are increased in ventricular hypertrophy associated with experimental hyperthyroidism. Since chronic administration of thyroxine (T4) improves postischaemic recovery in isolated heart models subjected to ischaemia-reperfusion stress, we tested the hypothesis that experimentally induced hyperthyroidism is associated with elevated expression of PTHrP and PTH.1R in rat myocardium. Hyperthyroid and control male Wistar rats were subjected to ischaemia-reperfusion stress using the Langendorff technique, and the PTHrP and PTH.1R expression was assessed by relative quantitative reverse transcriptase-polymerase chain reaction, Western blot analysis and immunohistochemistry. In the Langendorff model, the recovery of left ventricular developed pressure at the end of the stablization period and 45 min into the reperfusion period was used to assess the cardioprotective actions of T4 administration. Our data show that hyperthyroid animals had increased tolerance to the ischaemia-reperfusion stress and that this was associated with an increase of PTHrP and PTH.1R expression levels compared with those of control animals. In the control animals, the expression of PTHrP was increased 45 min into the reperfusion phase, while the PTH.1R expression pattern was significantly and gradually decreased throughout the ischaemia and reperfusion phases. In the hyperthyroid animals, the PTHrP and PTH.1R expression pattern was significantly higher throughout the ischaemia and reperfusion phases compared with that of control hearts. Our data suggest that increasing levels of PTHrP and PTH.1R expression can mediate, at least in part, the T4 administration-induced cardioprotection in rat ventricular myocardium. PMID- 17911358 TI - Sensory and autonomic nerve changes in the monosodium glutamate-treated rat: a model of type II diabetes. AB - Rats that had been injected with monosodium glutamate (MSG) neonatally were studied for up to 70 weeks and compared with age-matched control rats to study changes in glucose tolerance and in sympathetic and sensory nerves. At 61 and 65 weeks of age, there were significant differences in glucose tolerance between the MSG and control groups, and the MSG group had raised fasting blood glucose. These changes were not associated with changes in the number of beta-cells in the islets of Langerhans. In addition, the diabetic MSG-treated rats had central obesity and cataracts. Hypoalgesia to thermal stimuli was present in MSG-treated rats as early as 6 weeks and persisted at 70 weeks. However, no differences were observed in the distribution of substance P, the neurokinin-1 receptor or calcitonin gene-related peptide in the dorsal horn of L3-L5 at this age (70 weeks). Diabetic MSG-treated animals at 65 and 70 weeks of age had significantly reduced noradrenaline concentrations in the heart, tail artery and ileum, while concentrations in the adrenal gland and corpus cavernosum were significantly increased. There was also a significant increase in adrenal adrenaline, dopamine and serotonin, largely attributable to changes in weight of the adrenal gland in the MSG-treated animals. The results indicate that MSG-treated animals develop a form of type II diabetes by about 60 weeks of age, and that there are significant changes in amine levels in various tissues associated with these developments. PMID- 17911359 TI - Oxidative stress and cerebrovascular dysfunction in mouse models of Alzheimer's disease. AB - Several factors have been implicated in Alzheimer's disease (AD) but there is no definite conclusion as to the main pathogenic agents. Mutations in the amyloid precursor protein (APP) that lead to increased production of amyloid beta peptide (A beta) are associated with the early-onset, familial forms of AD. However, in addition to ageing, the most common risk factors for the sporadic, prevalent form of AD are hypertension, hypercholesterolaemia, ischaemic stroke, the ApoE4 allele and diabetes, all characterized by a vascular pathology. In AD, the vascular pathology includes accumulation of A beta in the vessel wall, vascular fibrosis, and other ultrastructural changes in constituent endothelial and smooth muscle cells. Moreover, the ensuing chronic cerebral hypoperfusion has been proposed as a determinant factor in the accompanying cognitive deficits. In transgenic mice that overexpress mutated forms of the human APP (APP mice), the increased production of A beta results in vascular oxidative stress and loss of vasodilatory function. The culprit molecule, superoxide, triggers the synthesis of other reactive oxygen species and the sequestration of nitric oxide (NO), thus impairing resting cerebrovascular tone and NO-dependent dilatations. The A beta induced cerebrovascular dysfunction can be completely abrogated in aged APP mice with antioxidant therapy. In contrast, in mice that overproduce an active form of the cytokine transforming growth factor-beta1 and recapitulate the vascular structural changes seen in AD, antioxidants have no beneficial effect on the accompanying cerebrovascular deficits. This review discusses the beneficial role and limitations of antioxidant therapy in AD cerebrovascular pathology. PMID- 17911360 TI - Biomarkers in geriatric psychiatry. PMID- 17911361 TI - The relationship between cerebrospinal fluid biomarkers and depression in elderly women. AB - OBJECTIVE: Cerebrospinal fluid (CSF) biomarkers including the 42 amino-acid form of beta-amyloid (Abeta42), total tau protein (T-tau), and the CSF/serum albumin ratio are markers of brain pathology and metabolism. Abeta42 and T-tau are sometimes used to discriminate geriatric depression from mild forms of Alzheimer disease (AD) in clinical studies. However, studies focusing on the relationship between these CSF biomarkers and geriatric depression are lacking. METHODS: This was a cross-sectional study with a population-based sample of 84 nondemented elderly women in Sweden. Measurements included neuropsychiatric, physical, and lumbar puncture examinations, with Diagnostic and Statistical Manual of Mental Disorders, Third Revision-based depression diagnoses and measurement of CSF levels of Abeta42, T-tau, albumin, and serum albumin. RESULTS: Fourteen women (mean age: 72.6 years) had any depression (11 with major depressive disorder [MDD]). Compared to women without depression, women with MDD had higher levels of Abeta42 and the CSF/serum albumin ratio. The CSF/serum albumin ratio was also higher in women with any depression. No differences in T-tau were observed; however, T-tau increased with age. CONCLUSION: Higher levels of CSF Abeta42 were observed among elderly depressed women, in contrast to lower levels usually observed in AD, indicating potential neuropathological differences between the two disorders. Higher CSF/serum albumin ratios observed in depressed women point to potential vascular processes. PMID- 17911362 TI - Allelic differences in the brain-derived neurotrophic factor Val66Met polymorphism in late-life depression. AB - OBJECTIVE: The Val66Met polymorphism of the brain-derived neurotrophic factor gene is associated with cognitive and neuroimaging changes. The authors examined the relationship between this polymorphism and depression in an elderly sample, hypothesizing that the Met66 allele would be associated with late-life depression. METHODS: A total of 245 elderly depressed white subjects and 94 elderly comparison white subjects completed clinical assessments and provided a blood sample for genotyping. Subjects were dichotomized as either homozygous for the Val66 allele or Met66 allele carriers. Gene frequencies were compared between groups, with separate analyses examining for differences in gene frequencies based on age of depression onset, family history, and depression history. Logistic regression models examined the relationship between genotype and depression after controlling for age, sex, and race. RESULTS: Depressed subjects were more likely to be Met66 allele carriers than were comparison subjects (38.8% versus 24.4%; chi(2) = 6.13, 1 df, p = 0.0133). This relationship remained significant after controlling for covariates (Wald chi(2) = 5.10, 1 df, p = 0.024; odds ratio: 1.92, 95% confidence interval: 1.09-3.38). There were no significant relationships between genotype and age of onset, number of episodes, or family history of depression. CONCLUSION: Met66 allele carriers have almost double the odds of having geriatric depression than do Val66 allele homozygotes. This polymorphism was unrelated to other clinical characteristics of depression in later life. PMID- 17911363 TI - ApoE4 allele is associated with late-life depression: a population-based study. AB - OBJECTIVE: This study aimed to clarify the relationship between the epsilon4 allele and late-life depression, taking into account lipid profile, vascular diseases, and sociodemographics. METHODS: Using a multilevel stratified random sampling strategy, a total of 500 subjects aged 65 to 74 years were recruited for this cross-sectional study from the official household records of an entire county in southern Taiwan. Depressive symptoms were assessed by the Taiwanese Depression Questionnaire (TDQ). Cognitive function was assessed by the Short Portable Mental Status Questionnaire. Blood samples were collected for the determination of the apolipoprotein E (ApoE) polymorphism and the lipid profile. RESULTS: A total of 283 subjects (58.7% male, with a mean age of 69.2 +/- 2.7 years) completed all questionnaires and collection of blood samples. Using the chi(2) test, the overall difference for frequency of the presence of the epsilon4 allele was significant among the severe group (TDQ score >18), moderate group (TDQ score 9-18), and mild group (TDQ score <9). The proportion of history of heart disease was significantly higher in the severe group than in the mild or moderate group. Kruskal-Wallis statistics revealed that the mean total and low density lipoprotein cholesterol levels were significantly higher in the severe group than in the moderate or mild group. With our two-level four-class latent class regression model, the presence of the epsilon4 allele was significantly associated with the severely depressed group as compared to the nondepressed group, adjusting for vascular diseases and lipid profile. CONCLUSION: The ApoE epsilon4 allele may be correlated with severe depression in the elderly through ways other than the "vascular depression" hypothesis. PMID- 17911364 TI - Apolipoprotein E epsilon4 genotype and gender: effects on memory. AB - OBJECTIVE: Episodic recognition memory for odors and visual was assessed in apolipoprotein E (ApoE) epsilon4-positive and epsilon4-negative men and women diagnosed with Alzheimer disease (AD) and a healthy age- and gender-matched comparison group. METHODS: A total of 38 AD patients and 38 age- and gender matched healthy older adults completed a recognition memory task involving three categories of stimuli: odors, faces, and symbols. RESULTS: In the healthy comparison group, men who were epsilon4 negative outperformed epsilon4-positive men in recognition memory for odors and committed fewer false-positive errors. However, there were no significant differences between epsilon4-negative and epsilon4-positive women in the comparison group. No significant gender or ApoE status differences were detected in recognition memory for faces or symbols in the comparison group. In patients with AD, epsilon4-negative women outperformed epsilon4-positive women in recognition memory for odors and committed significantly fewer false-positive errors. However, there were no significant differences between epsilon4-positive and epsilon4-negative men. There were no significant gender or ApoE status differences in recognition memory for faces or symbols in AD patients. CONCLUSION: The results demonstrate that recognition memory for olfactory stimuli may be particularly impaired in healthy older men with the epsilon4 allele. In patients with AD, odor memory impairments may be less severe in women who are negative for the epsilon4 allele. The results offer new insight into how recognition memory is affected by gender, the epsilon4 allele, and the modality of the stimulus to be remembered in healthy older adults and patients with AD. PMID- 17911365 TI - Glutathione S-transferase P1 and T1 gene polymorphisms predict longitudinal course and age at onset of Alzheimer disease. AB - OBJECTIVE: Oxidative stress has been suggested as a contributor of Alzheimer disease (AD) neurodegeneration, particularly in those patients with late-onset AD (LOAD). Therefore, the authors studied the effect of glutathione S-transferase (GST) P1-M1-T1 gene polymorphisms and their interactions with the apolipoprotein E (ApoE) epsilon4 allelic variant on the three-year longitudinal course of AD. METHODS: Global cognitive level as measured by the Mini-Mental State Exam, basic activities of daily living (BADLs) as measured by the Physical Self-Maintenance Scale, and behavior as measured by the Neuropsychiatric Inventory, were assessed at baseline and after 1, 2, and 3 years in a sample of 99 LOAD patients. These subjects were drug naive and had undergone the first clinical examination for the diagnosis of AD. RESULTS: A multiple regression analysis indicated that the presence of ApoE epsilon4 allelic variant or GSTT1 null phenotype predicted the faster age at onset of the illness (F = 5.76, df = 2, 96, p = 0.0043). Carriers of GSTP1 *C allelic variant had a faster decline in cognitive functions (repeated measures analysis of variance [ANOVA]: F = 4.00, df = 3, 285, p = 0.008) and in BADLs (repeated measures ANOVA: F = 5.27, df = 3, 285, p = 0.001). This faster decline was independent from ApoE epsilon4 allele possession. No effect of GST P1 M1-T1 polymorphisms was found on behavioral symptom severity. CONCLUSION: These data are in line with the hypotheses that oxidative damage is a prominent feature in the clinical progression and the age at onset of LOAD. PMID- 17911366 TI - Relationship between the Hamilton Depression Rating Scale and the Montgomery Asberg Depression Rating Scale in depressed elderly: a meta-analysis. AB - OBJECTIVE: To determine whether the sensitivity of the Hamilton Depression Rating Scale (HDRS) and Montgomery-Asberg Depression Rating Scale (MADRS) to treatment effects are comparable in geriatric antidepressant randomized controlled trials by developing and validating an equation that links between the two instruments. METHODS: Literature search for this meta-analysis was based on three sources: MEDLINE, a recent related meta-analysis, and experts in geriatric antidepressant trials. The search resulted in 11 relevant geriatric antidepressant trial studies that administered both instruments for symptom ratings. The authors used baseline ratings as a model-building sample and postrandomization ratings as a validation sample. HDRS scores were prorated into HDRS17, a 17-item HDRS, for analysis. The development and validation was based on a total number of 1,874 subjects. RESULTS: The correlations were high between baseline mean HDRS17 and MADRS ratings (r = 0.80; Fisher's z = 1.09, N = 25, p <0.0001) and between postrandomization ratings (r = 0.88, Fisher's z = 1.39, N = 65, p <0.0001). The following equation was derived: HDRS17 = -1.58 + 0.86 x MADRS. The difference between observed and estimated HDRS17 in a validation sample consisting of postrandomization follow-up means did not depend on magnitudes of HDRS17. CONCLUSION: Although generalizability of findings into a broader population could be limited, and the authors could not assess concordance of changes of particular item constructs between HDRS and MADRS ratings, both ratings are comparable in assessing changes in overall depressive symptom severity in response to antidepressants in depressed elderly at aggregated group mean levels. PMID- 17911367 TI - Integration of mental health services into primary care overcomes ethnic disparities in access to mental health services between black and white elderly. AB - OBJECTIVE: The authors evaluated whether the integration of mental health into primary care overcomes ethnic disparities in access to and participation in mental health (MH) and substance abuse (SA) treatment. METHODS: The authors conducted site-specific analysis of a multisite clinical trial to compare participation of black and white elderly in an integrated model of care (all MH/SA services are provided at primary care clinics) versus an enhanced referral model of care (all MH/SA services are provided at specialized MH clinics). In all, 183 elderly (56% black) diagnosed with depression (82%), anxiety (32%), and/or problem drinking (22%) were randomized. RESULTS: Blacks in the integrated arm were significantly more likely to have at least one MH/SA visit (77.5%) relative to blacks in the enhanced referral arm (22%; adjusted odds ratio [OR]: 14.13; confidence interval [CI]: 4.76-41.95, Wald chi(2): 22.75, df = 1, p <0.0001). There was no statistically significant difference between whites in the integrated treatment arm (66.6%) and whites in the enhanced referral arm (46.9%, adjusted OR: 2.98; CI: 0.98-9.06, Wald chi(2): 3.72, df = 1, p = 0.05). In the enhanced referral arm, blacks had a significantly smaller number of overall MH/SA visits (mean [SD]: 2.08 [5.28]) relative to whites (mean [SD]: 5.31 [7.76], adjusted incident rate ratio [IRR]: 2.87; CI: 1.06-7.73, Wald chi(2): 4.37, df = 1, p = 0.03). In the integrated arm, there was no statistically significant difference between blacks (mean [SD]: 3.22 [3.71]) and whites (mean [SD]: 2.75 [4.29], adjusted IRR: 0.58; CI: 0.25-1.33, Wald chi(2): 1.64, df = 1, p = 0.20). For both groups, time between baseline evaluation to first MH/SA visit was significantly shorter in the integrated treatment arm (for blacks: mean days [SD]: 31.06 [28.66]; for whites: mean days [SD]: 22.18 [33.88]) than in the enhanced referral arm (mean [SD]: 62.45 [43.53], adjusted hazard ratio [HR]: 7.82; CI: 3.65-16.75, Wald chi(2): 28.02, df = 1, p <0.0001; mean [SD]: 63.46 [32.41], adjusted HR: 2.48; CI: 1.20-5.13, Wald chi(2): 6.02, df = 1, p = 0.01, respectively). CONCLUSION: An integrated model of care is particularly effective in improving access to and participation in MH/SA treatment among black primary care patients. PMID- 17911368 TI - Tilt testing potentiated with sublingual nitroglycerin in children with unexplained syncope. AB - AIMS: The aim of this prospective study was to assess the diagnostic value of a sublingual nitroglycerin (NTG) potentiated head-up tilt (HUT) testing protocol in children with unexplained syncope. METHODS AND RESULTS: One hundred and sixty four consecutive paediatric patients with syncope of unknown origin and no evidence of organic heart disease (115 female, mean age 13 +/- 3 years) and 29 control children underwent a sublingual NTG-potentiated tilt testing protocol. Paediatric patients and controls were tilted at 60 degrees for 20 min and, if no symptom occurred, for other 15 min after sublingual 400 mug spray NTG administration. During the drug-free phase of the test, 13 patients (8%) and no controls had a positive response. After drug administration, a positive response occurred in another 88 patients (55%) and in four controls (14%), whereas an exaggerated response was observed in nine patients (5%) and in four controls (14%). Thus, the positive rate and specificity of sublingual NTG HUT test in children were 63 and 86%, respectively. No relevant adverse events were observed during the test. CONCLUSION: Tilt testing potentiated with sublingual NTG has proved to be effective and safe in unmasking the neurally mediated origin of unexplained syncope in children. The NTG challenge greatly increased the positive rate of passive tilt, with a small decrease in specificity. PMID- 17911369 TI - Time of day accounts for overnight improvement in sequence learning. AB - The theory that certain skills improve with a night of sleep has received considerable interest in recent years. However, because sleep typically occurs at the same time of day in humans, it is difficult to separate the effects of sleep from those of time of day. By using a version of the Serial Response Time Task, we assessed the role of sleep in implicit sequence learning while controlling for possible time-of-day effects. We replicated the apparent benefit of sleep on human participants. However, our data show that sleep does not affect implicit sequence learning; rather, time of day affects the ability of participants to express what they have learned. PMID- 17911370 TI - Glucocorticoid administration into the dorsal striatum [corrected] facilitates memory consolidation of inhibitory avoidance training but not of the context or footshock components. AB - It is well established that glucocorticoid administration into a variety of brain regions facilitates memory consolidation of fear-conditioning tasks, including inhibitory avoidance. The present findings indicate that the natural glucocorticoid corticosterone administered into the dorsal striatum (i.e., caudate nucleus) of male Wistar rats produced dose- and time-dependent enhancement of inhibitory avoidance memory consolidation. However, as assessed with a modified inhibitory avoidance procedure that took place on two sequential days to separate context training from footshock training, corticosterone administration into the dorsal striatum did not enhance memory of either the contextual or aversively motivational aspects of the task. PMID- 17911371 TI - Forward and backward second-order Pavlovian conditioning in honeybees. AB - Second-order conditioning (SOC) is the association of a neutral stimulus with another stimulus that had previously been combined with an unconditioned stimulus (US). We used classical conditioning of the proboscis extension response (PER) in honeybees (Apis mellifera) with odors (CS) and sugar (US). Previous SOC experiments in bees were inconclusive, and, therefore, we attempted to demonstrate SOC in the following three experiments: (Experiment 1) After differential conditioning (pairing odor A with US and presenting odor B without US), the bees experienced two pairs of partially overlapping odors, either a new odor C followed by a previously reinforced odor A (C-A) or a new odor C followed by a previously nonreinforced odor B (C-B). (Experiment 2) After differential conditioning, bees were presented with C-A or A-C. (Experiment 3) Bees were first presented with C-A or A-C before differential conditioning and were tested with odor C. We observed: (Experiment 1) 40% of the bees showed PER to the C-A presentation, but only 20% showed PER to the C-B presentation. (Experiment 2) 40% of the bees showed PER to the C-A presentation, while only 20% showed PER to the reversed sequence A-C. Experiments 1 and 2 showed that a previously reinforced odor can be a secondary reinforcer for excitatory SOC only with forward-pairing. (Experiment 3) PER toward C was lower (15%) in bees presented with A-C than with C-A (25%). This showed that backward SOC is not as effective as forward SOC. These results help to delineate different conditions that are critical for the phenomenon of SOC. PMID- 17911372 TI - The neural correlates of conceptual and perceptual false recognition. AB - False recognition, broadly defined as a claim to remember something that was not encountered previously, can arise for multiple reasons. For instance, a distinction can be made between conceptual false recognition (i.e., false alarms resulting from semantic or associative similarities between studied and tested items) and perceptual false recognition (i.e., false alarms resulting from physical similarities between studied and tested items). Although false recognition has been associated with frontal cortex activity, it is unclear whether this frontal activity can be modulated by the precise relationship between studied and falsely remembered items. We used event-related fMRI to examine the neural basis of conceptual compared with perceptual false recognition. Results revealed preferential activity in multiple frontal cortex regions during conceptual false recognition, which likely reflected increased semantic processing during conceptual (but not perceptual) memory errors. These results extend recent reports that different types of false recognition can rely on dissociable neural substrates, and they indicate that the frontal activity that is often observed during false compared with true recognition can be modulated by the relationship between studied and tested items. PMID- 17911374 TI - Acetylbritannilactone Inhibits Neointimal Hyperplasia after Balloon Injury of Rat Artery by Suppressing Nuclear Factor-{kappa}B Activation. AB - Based on our previous observations that 1-O-acetylbritannilactone (R) 4((3aS,4S,7aR)-4-hydroxy-6-methyl-3-methylene-2-oxo-2,3,3a,4,7,7a hexahydrobenzofuran-5-yl)pentyl acetate (ABL) suppresses prostaglandin E(2) and nitric oxide synthesis in macrophages, the present study was designed to explore the effect of ABL on neointimal hyperplasia after balloon injury and its mechanism of action. In male Sprague-Dawley rats, 26 mg/kg ABL or polyglycol (control) was administered daily from 3 days before injury to 2 weeks after conventional balloon injury. ABL administration led to a significant reduction in neointimal formation (neointima to media ratio, 1.94 +/- 0.43 versus 0.84 +/- 0.29, P < 0.01) and proliferative activity of vascular smooth muscle cells after balloon injury in rats. Western blot analysis revealed that this is correlated to the inhibition of nuclear factor (NF)-kappaB activation and to the reduced expression of cyclooxygenase-2. Investigation of potential signaling pathways demonstrated that ABL inhibited NF-kappaB activation via the blockade of the inhibitor of NF-kappaB kinase-beta activation and the suppression of the degradation of the inhibitors of NF-kappaB-alpha. These findings suggest that ABL is a potential inhibitor of neointimal formation because it blocks injury-induced NF-kappaB activation and may have beneficial effects in reducing the risk of restenosis after angioplasty. PMID- 17911375 TI - Immune modulatory treatment of trinitrobenzene sulfonic acid colitis with calcitriol is associated with a change of a T helper (Th) 1/Th17 to a Th2 and regulatory T cell profile. AB - A number of recent studies testify that calcitriol alone or in combination with corticosteroids exerts strong immune modulatory activity. As a new approach, we evaluated the protolerogenic potential of calcitriol and dexamethasone in acute T helper (Th)1-mediated colitis in mice. A rectal enema of trinitrobenzene sulfonic acid (TNBS) (100 mg/kg) was applied to BALB/c mice. Calcitriol and/or dexamethasone were administered i.p. from days 0 to 3 or 3 to 5 following the instillation of the haptenating agent. Assessment of colitis severity was performed daily. Colon tissue was analyzed macroscopically and microscopically, and myeloperoxidase activity, as well as cytokine levels [tumor necrosis factor alpha, interferon-gamma, interleukin (IL)-12p70, IL-1beta, IL-10, IL-4] were determined by enzyme-linked immunosorbent assay, T-bet, GATA family of transcription factors 3, a Th2 master regulator (GATA3), Foxp3, cytotoxic T lymphocyte-associated antigen 4 (CTLA4), IL-23p19 and IL-17 expression by immunoblot analysis. The combination of the steroids most effectively reduced the clinical and histopathologic severity of TNBS colitis. Th1-related parameters were down-regulated, whereas Th2 markers like IL-4 and GATA3 were up-regulated. Apart from known steroid effects, calcitriol in particular promoted regulatory T cell profiles as indicated by a marked increase of IL-10, TGFbeta, FoxP3, and CTLA4. Furthermore, analysis of dendritic cell mediators responsible for a proinflammatory differentiation of T cells revealed a significant reduction of IL 12p70 and IL23p19 as well as IL-6 and IL-17. Thus, our data support a rationale for a steroid-sparing, clinical application of calcitriol derivatives in inflammatory bowel disease. Furthermore they suggest that early markers of inflammatory dendritic cell and Th17 differentiation qualify as new target molecules for both calcitriol and highly selective immune-modulating vitamin D analogs. PMID- 17911373 TI - Sex-dependent up-regulation of two splicing factors, Psf and Srp20, during hippocampal memory formation. AB - Gene transcription is required for long-term memory (LTM) formation. LTM formation is impaired in a male-specific manner in mice lacking either of the two Ca(2+)/calmodulin-dependent kinase kinase (Camkk) genes. Since altered transcription was suggested to cause these impairments in LTM formation, we used microarrays to screen for CaMKKbeta-dependent gene expression changes. Here we show that the hippocampal mRNA expression of two splicing factors, splicing factor arginine/serine-rich 3 (Sfrs3/Srp20) and polypyrimidine tract-binding protein-associated splicing factor (Psf), is altered in CaMKKbeta-deficient males. In wild-type (WT) mice, the basal expression level in the hippocampus is higher in males than in females, and the sex difference in Srp20 expression is detectable before puberty. Training in two hippocampus-dependent learning tasks, the spatial version of the Morris water maze (MWM) and background contextual fear conditioning, increases the hippocampal mRNA expression of both splicing factors in WT males. However, the increase in Srp20 mRNA expression occurs only in males and not in females, whereas the up-regulation of Psf expression occurs in both sexes. Importantly, control experiments demonstrate that the up-regulation of both splicing factors is specific for the learned associations after contextual fear conditioning. In summary, we provide the first evidence for a regulation of splicing factors during LTM formation and we suggest that alternative splicing contributes to sex differences in LTM formation. PMID- 17911376 TI - Angiotensin II activates extracellular signal-regulated kinase independently of receptor tyrosine kinases in renal smooth muscle cells: implications for blood pressure regulation. AB - Angiotensin II can cause hypertension through enhanced vasoconstriction of renal vasculature. One proposed mechanism for reduction of angiotensin II-induced hypertension is through inhibition of the mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase cascade. MEK/ERK has been shown to phosphorylate the regulatory subunit of myosin light chain at identical positions as myosin light chain kinase. There are multiple mechanisms proposed regarding angiotensin II-mediated ERK activation. We hypothesized that renal microvascular smooth muscle cells (RmuVSMCs) signal through a unique pathway compared with thoracic aorta smooth muscle cells (TASMCs), which is involved in blood pressure regulation. Use of epidermal growth factor (EGF) and platelet derived growth factor (PDGF) receptor-specific inhibitors 4-(3-chloroanilino)-6,7-dimethoxyquinazoline (AG1478) and 6,7 dimethoxy-3-phenylquinoxaline (AG1296), respectively, demonstrates that angiotensin II activates ERK in TASMCs, but not RmuVSMCs, through transactivation of EGF and PDGF receptors. In addition, inhibition of Src with its specific inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo[3,4-d]pyrimidine (PP2) abolishes angiotensin II-, but not EGF-or PDGF-, mediated phosphorylation of ERK in RmuVSMCs, yet it has no effect in TASMCs. The physiological significance of transactivation was examined in vivo using anesthetized Wistar-Kyoto rats with 15 mg/kg 2'-amino-3'-methoxyflavone (PD98059), an MEK inhibitor, as well as 20 mg/kg AG1478 and 1.5 mg/kg AG1296 in an acute model of angiotensin II-mediated increase in blood pressure. None of the inhibitors had an effect on basal blood pressure, and only PD98059 reduced angiotensin II-mediated increase in blood pressure. Moreover, in RmuVSMCs, but not TASMCs, angiotensin II localizes phosphorylated ERK to actin filaments. In conclusion, angiotensin II signals through a unique mechanism in the renal vascular bed that may contribute to hypertension. PMID- 17911377 TI - Bone scanning--who needs it among patients with newly diagnosed prostate cancer? AB - BACKGROUND: We evaluated the relationship between serum PSA and clinical variables to eliminate bone scanning in patients with prostate cancer having a low probability of bone metastasis. METHODS: The study included 366 patients with newly diagnosed prostate cancer between 1999 and 2005. Bone metastasis was studied for its correlation with various clinical and pathological variables in these patients. RESULTS: Bone metastasis was found in 28 (7.7%) of 366 patients. Fourteen patients had skeletal symptoms related to bone metastasis. The risk for bone metastases increased considerably with increases of PSA level, clinical T stage and Gleason score. The metastasis was not found in 161 patients with serum PSA concentration of 10 ng/ml or lower. In 95 patients with the concentration between 10 and 20 ng/ml only two had the metastasis. These two patients had T2 disease and Gleason scores of 7 or greater. In 204 patients with clinical stage T1 disease, one (0.5%) had the metastasis. In 117 patients with Gleason scores of 6 or less, the metastasis was found in two (1.7%). CONCLUSIONS: For patients with serum PSA levels of 10 ng/ml or lower, bone scanning may be eliminated because of the negligible risk of bone metastases. In addition, scanning may not be necessary for those with PSA levels between 10 and 20 ng/ml, when they have T1 disease and Gleason scores of 6 or lower. PMID- 17911379 TI - Microarray analysis reveals distinctive signaling between the bed nucleus of the stria terminalis, nucleus accumbens, and dorsal striatum. AB - To identify distinct transcriptional patterns between the major subcortical dopamine targets commonly studied in addiction we studied differences in gene expression between the bed nucleus of the stria terminalis (BNST), nucleus accumbens (NAc), and dorsal striatum (dStr) using microarray analysis. We first tested for differences in expression of genes encoding transcripts for common neurotransmitter systems as well as calcium binding proteins routinely used in neuroanatomical delineation of brain regions. This a priori method revealed differential expression of corticotropin releasing hormone (Crh), the GABA transporter (Slc6a1), and prodynorphin (Pdyn) mRNAs as well as several others. Using a gene ontology tool, functional scoring analysis, and Ingenuity Pathway Analysis, we further identified several physiological pathways that were distinct among these brain regions. These two different analyses both identified calcium signaling, G-coupled protein receptor signaling, and adenylate cyclase-related signaling as significantly different among the BNST, NAc, and dStr. These types of signaling pathways play important roles in, amongst other things, synaptic plasticity. Investigation of differential gene expression revealed several instances that may provide insight into reported differences in synaptic plasticity between these brain regions. The results support other studies suggesting that crucial pathways involved in neurotransmission are distinct among the BNST, NAc, and dStr and provide insight into the potential use of pharmacological agents that may target region-specific signaling pathways. Furthermore, these studies provide a framework for future mouse-mouse comparisons of transcriptional profiles after behavioral/pharmacological manipulation. PMID- 17911378 TI - Patterns of care study of breast-conserving therapy in Japan: comparison of the treatment process between 1995 1997 and 1999 2001 surveys. AB - BACKGROUND: The Japan Patterns of Care Study (JPCS) conducted two national surveys to identify changes associated with the treatment process of care for patients undergoing breast-conserving therapy (BCT). Between the two national surveys, the Japanese Breast Cancer Society published its treatment guideline for BCT. METHOD: The first survey collected data on 865 patients treated between 1995 and 1997 (JPCS-1), and the second on 746 patients treated between1999 and 2001 (JPCS-2) by extramural audits. RESULTS: There was a shift to an older age distribution in JPCS-2 compared with JPCS-1. In JPCS-2, the average patient age was 53.9 compared with 51.5 in JPCS-1 (P < 0.001). There was a reduction in the extent of breast surgery and the proportion of the patients who received quadrantectomy was 57.0% in JPCS-1 and 30.3% in JPCS-2 (P < 0.001). In JPCS-2, a cast or shell for immobilization was used at a significantly higher rate of 52.9% compared with 32.6% for JPCS-1 (P < 0.001). The rate of boost irradiation was increased in JPCS-2, especially for patients with a positive surgical margin; it was significantly increased to 83.5% in JPCS-2 compared with 53.9% in JPCS-1 (P < 0.001). CONCLUSIONS: The second survey revealed a rapid change in the trend of the treatment of BCT in Japan and represented high compliance of the treatment guideline for BCT published by the Japanese Breast Cancer Society (JBCS) in 1999. PMID- 17911380 TI - Alternative promoter usage and alternative splicing contribute to mRNA heterogeneity of mouse monocarboxylate transporter 2. AB - Expression patterns of monocarboxylate transporter 2 (MCT2) display mRNA diversity in a tissue-specific fashion. We cloned and characterized multiple mct2 5'-cDNA ends from the mouse and determined the structural organization of the mct2 gene. We found that transcription of this gene was initiated from five independent genomic regions that spanned >80 kb on chromosome 10, resulting in five unique exon 1 variants (exons 1a, 1b, 1c, 1d, and 1e) that were then spliced to the common exon 2. Alternative splicing of four internal exons (exons AS1, AS2, AS3, and exon 3) greatly increased the complexity of mRNA diversity. While exon 1c was relatively commonly used for transcription initiation in various tissues, other exon 1 variants were used in a tissue-specific fashion, especially exons 1b and 1d that were used exclusively for testis-specific expression. Sequence analysis of 5'-flanking regions upstream of exons 1a, 1b, and 1c revealed the presence of numerous potential binding sites for ubiquitous transcription factors in all three regions and for transcription factors implicated in testis-specific or hypoxia-induced gene expression in the 1b region. Transient transfection assays demonstrated that each of the three regions contained a functional promoter and that the in vitro, cell type-specific activities of these promoters were consistent with the tissue-specific expression pattern of the mct2 gene in vivo. These results indicate that tissue-specific expression of the mct2 gene is controlled by multiple alternative promoters and that both alternative promoter usage and alternative splicing contribute to the remarkable mRNA diversity of the gene. PMID- 17911382 TI - Gene expression profiling in lung fibroblasts reveals new players in alveolarization. AB - Little is known about the molecular basis of lung alveolarization. We used a microarray profiling strategy to identify novel genes that may regulate the secondary septation process. Rat lung fibroblasts were extemporaneously isolated on postnatal days 2, 7, and 21, i.e., before, during, and after septation, respectively. Total RNA was extracted, and cRNAs were hybridized to Affymetrix rat genome 230 2.0 microarrays. Expression levels of a selection of genes were confirmed by real-time PCR. In addition to genes already known to be upregulated during alveolarization including drebrin, midkine, Fgfr3, and Fgfr4, the study allowed us to identify two remarkable groups of genes with opposite profiles, i.e., gathering genes either transiently up- or downregulated on day 7. The former group includes the transcription factors retinoic acid receptor (RXR) gamma and homeobox (Hox) a2, a4, and a5 and genes involved in Wnt signaling (Wnt5a, Fzd1, and Ndp); the latter group includes the extracellular matrix components Comp and Opn and the signal molecule Slfn4. Profiling in whole lung from fetal life to adulthood confirmed that changes were specific for alveolarization. Two treatments that arrest septation, hyperoxia and dexamethasone, inhibited the expression of genes that are upregulated during alveolarization and conversely enhanced that of genes weakly expressed during alveolarization and upregulated thereafter. The possible roles of these genes in secondary septation are discussed. Gene expression profiling analysis on freshly isolated cells represents a powerful approach to provide new information about differential regulation of genes during alveolarization and pathways potentially involved in the pathogenesis of bronchopulmonary dysplasia. PMID- 17911381 TI - Allelic variation of the Tas1r3 taste receptor gene selectively affects taste responses to sweeteners: evidence from 129.B6-Tas1r3 congenic mice. AB - The Tas1r3 gene encodes the T1R3 receptor protein, which is involved in sweet taste transduction. To characterize ligand specificity of the T1R3 receptor and the genetic architecture of sweet taste responsiveness, we analyzed taste responses of 129.B6-Tas1r3 congenic mice to a variety of chemically diverse sweeteners and glucose polymers with three different measures: consumption in 48 h two-bottle preference tests, initial licking responses, and responses of the chorda tympani nerve. The results were generally consistent across the three measures. Allelic variation of the Tas1r3 gene influenced taste responsiveness to nonnutritive sweeteners (saccharin, acesulfame-K, sucralose, SC-45647), sugars (sucrose, maltose, glucose, fructose), sugar alcohols (erythritol, sorbitol), and some amino acids (D-tryptophan, D-phenylalanine, L-proline). Tas1r3 genotype did not affect taste responses to several sweet-tasting amino acids (L-glutamine, L threonine, L-alanine, glycine), glucose polymers (Polycose, maltooligosaccharide), and nonsweet NaCl, HCl, quinine, monosodium glutamate, and inosine 5'-monophosphate. Thus Tas1r3 polymorphisms affect taste responses to many nutritive and nonnutritive sweeteners (all of which must interact with a taste receptor involving T1R3), but not to all carbohydrates and amino acids. In addition, we found that the genetic architecture of sweet taste responsiveness changes depending on the measure of taste response and the intensity of the sweet taste stimulus. Variation in the T1R3 receptor influenced peripheral taste responsiveness over a wide range of sweetener concentrations, but behavioral responses to higher concentrations of some sweeteners increasingly depended on mechanisms that could override input from the peripheral taste system. PMID- 17911383 TI - Re: Antiphospholipid antibodies in serum and follicular fluid: is there a correlation with IVF implantation failure? PMID- 17911384 TI - ALK5- and TGFBR2-independent role of ALK1 in the pathogenesis of hereditary hemorrhagic telangiectasia type 2. AB - ALK1 belongs to the type I receptor family for transforming growth factor-beta family ligands. Heterozygous ALK1 mutations cause hereditary hemorrhagic telangiectasia type 2 (HHT2), a multisystemic vascular disorder. Based largely on in vitro studies, TGF-beta1 has been considered as the most likely ALK1 ligand related to HHT, yet the identity of the physiologic ALK1 ligand remains controversial. In cultured endothelial cells, ALK1 and another TGF-beta type I receptor, ALK5, regulate angiogenesis by controlling TGF-beta signal transduction, and ALK5 is required for ALK1 signaling. However, the extent to which such interactions between these 2 receptors play a role in pathogenesis of HHT is unknown. We directly addressed these issues in vivo by comparing the phenotypes of mice in which the Alk1, Alk5, or Tgfbr2 gene was conditionally deleted in restricted vascular endothelia using a novel endothelial Cre transgenic line. Alk1-conditional deletion resulted in severe vascular malformations mimicking all pathologic features of HHT. Yet Alk5- or Tgfbr2 conditional deletion in mice, or Alk5 inhibition in zebrafish, did not affect vessel morphogenesis. These data indicate that neither ALK5 nor TGFBR2 is required for ALK1 signaling pertinent to the pathogenesis of HHT and suggest that HHT might not be a TGF-beta subfamily disease. PMID- 17911385 TI - Microvascular endothelial cells express a phosphatidylserine receptor: a functionally active receptor for phosphatidylserine-positive erythrocytes. AB - Phosphatidylserine (PS)-positive erythrocytes adhere to endothelium and subendothelial matrix components. While thrombospondin mediates these inter actions, it is unknown whether PS-associated erythrocyte-endothelial adhesion occurs in the absence of plasma ligands. Using ionophore-treated PS-expressing control HbAA erythrocytes, we demonstrate that PS-positive erythrocytes adhered to human lung microendothelial cells in the absence of plasma ligands, that this adhesion was enhanced following endothelial activation with IL-1alpha, TNF-alpha, LPS, hypoxia, and heme, and that this adhesive interaction was selective to erythrocyte PS. We next explored whether microendothelial cells express an adhesion receptor that recognizes cell surface-expressed PS (PSR) similar to that expressed on activated macrophages. We demonstrate constitutive expression of both PSR mRNA and protein that were up-regulated in a time-dependent manner following endothelial activation. While minimal PSR expression was noted on unstimulated cells, endothelial activation up-regulated PSR surface expression. In antibody-blocking studies, using PS-positive erythrocytes generated either artificially via ionophore treatment of control erythrocytes or from patients with sickle cell disease, we demonstrate that PSR was functional, supporting PS mediated erythrocyte adhesion to activated endothelium. Our results demonstrate the existence of a novel functional adhesion receptor for PS on the microendothelium that is up-regulated by such pathologically relevant agonists as hypoxia, cytokines, and heme. PMID- 17911387 TI - Unliganded estrogen receptor alpha inhibits breast cancer cell growth through interaction with a cyclin-dependent kinase inhibitor (p21(WAF1)). AB - Estrogens are mitogenic in human breast cancer cells, but the presence of estrogen receptor alpha (ER alpha) is associated with a favorable prognosis in primary tumors and the molecular basis for this paradoxical relationship remains unknown. Here we show that ER alpha and ER alpha mutants devoid of ligand and DNA binding domains inhibit cell growth in three-dimensional matrix as well as tumor formation in nude mice. Using in vitro and intracellular approaches, we have found that ER alpha, via its amino acids 184-283, interacts with cyclin-dependent kinase inhibitor p21(WAF1). Both proteins exhibit mutual interactions in the absence of estrogens or in the presence of pure antiestrogen ICI(182,780), whereas estradiol treatment disrupts their interactions. Cross-linking experiments reveal that these proteins are present in a larger complex of approximately 200 kDa that also contains cdk2 and cyclin E. We further demonstrate that the unliganded full-length ER alpha or the variant having the p21(WAF1) interaction region significantly increases p21(WAF1) expression, whereas ER alpha silencing reduces p21(WAF1) levels and silencing of p21(WAF1) is sufficient to prevent ER alpha-induced growth inhibition. Taken together, our results point to an antiproliferative function of the unliganded ER alpha through its physical interactions with p21(WAF1) that may also explain the favorable prognosis of ER alpha-positive breast cancers. PMID- 17911386 TI - Increased risk of breast cancer among survivors of allogeneic hematopoietic cell transplantation: a report from the FHCRC and the EBMT-Late Effect Working Party. AB - As risk for secondary breast cancer is elevated among cancer survivors treated with conventional therapy, we sought to determine the risk among 3337 female 5 year survivors who underwent an allogeneic hematopoietic cell transplantation (HCT) at the Fred Hutchinson Cancer Research Center or at one of 82 centers reporting to the European Bone Marrow Transplant Registry. Risk was calculated using standardized incidence ratios (SIRs), and risk factors were evaluated with a multivariable Cox proportional hazards model. Fifty-two survivors developed breast cancer at a median of 12.5 (range: 5.7-24.8) years following HCT (SIR=2.2). Twenty-five-year cumulative incidence was 11.0%, higher among survivors who received total body irradiation (TBI) (17%) than those who did not receive TBI (3%). In multivariable analysis, increased risk was associated with longer time since transplantation (hazard ratio [HR] for 20+ years after transplantation=10.8), use of TBI (HR=4.0), and younger age at transplantation (HR=9.5 for HCT<18 years). Hazard for death associated with breast cancer was 2.5 (95% CI: 1.1-5.8). We conclude that female survivors of allogeneic HCT are at increased risk of breast cancer and should be educated about the need for regular screening. PMID- 17911388 TI - Impact of baseline protease genotype and phenotype on the response to darunavir outside clinical trials. PMID- 17911389 TI - Spontaneous mutation frequency and emergence of ciprofloxacin resistance in Campylobacter jejuni and Campylobacter coli. AB - OBJECTIVES: We analysed the contribution of spontaneous mutation frequency to the evolution of ciprofloxacin resistance and the diversity of mutations in the quinolone resistance-determining region (QRDR) of gyrA and in the intergenic region, cmeR-cmeA, of the CmeABC efflux pump in Campylobacter jejuni and Campylobacter coli. METHODS: The mutation frequency was measured in 11 quinolone susceptible C. jejuni and 5 C. coli strains, with and without ox bile. MICs and target-specific and efflux-associated mutations were studied for a number of colonies of each strain selected from plates containing 1 mg/L ciprofloxacin. RESULTS: The spontaneous mutation frequency level among susceptible C. jejuni and C. coli strains ranged from hypomutable (approximately 4 x 10(-9)) to strongly mutable (approximately 7 x 10(-3)). Ox bile had no effect on mutation frequency. Pre-existing ampicillin and tetracycline resistance increased the MICs of ciprofloxacin for two strains from 0.032-0.125 to 0.5 mg/L. MICs of ciprofloxacin for the colonies selected from plates containing 1 mg/L ciprofloxacin varied from 1 to 16 mg/L, with the Thr-86-->Ile or Asp-90-->Asn mutation detected in the QRDR of gyrA. In 21.5% (14/65) of the selected colonies, no specific mutations existed. Two types of mutations in CmeR promoter-binding inverted sequences were identified both in the parent strains and in the colonies selected from ciprofloxacin plates. CONCLUSIONS: The variation in mutation frequencies between most C. jejuni and C. coli strains was up to 700-fold. Mutation in the QRDR of gyrA or in the intergenic region was not associated with differences in spontaneous mutation frequencies. Previously acquired resistance to tetracycline and ampicillin predisposed strains to high-level ciprofloxacin resistance and to multiple antibiotic resistance. PMID- 17911390 TI - Meeting report: Messages from Moscow--the BSAC at the ninth IACMAC International Congress on Antimicrobial Therapy. AB - The organizing committee of the Russian Interregional Association for Clinical Microbiology and Antimicrobial Chemotherapy invited an international delegation of representatives, including nine BSAC members, to contribute to their ninth congress in Moscow at the end of May 2007. The event demonstrated the value of comparing experience in the context of an educational meeting and how efficient organization can overcome potential communication problems when simultaneous audio and slide translations are used to best effect. PMID- 17911391 TI - Diamonds are forever: the cortisone legacy. AB - The year 1946 was not only the year that the Society for Endocrinology was founded, but also the year that Edward Kendall's compound E (cortisone) was first synthesised by Louis Sarett. By 1948, sufficient quantities of compound E were available for the rheumatologist Philip Hench to test it successfully for the first time in a patient with rheumatoid arthritis. It was immediately hailed as a 'wonder drug' and was shown to be effective in a number of inflammation associated conditions, most notably rheumatoid arthritis. The subsequent development of endocrinology as a discipline is inextricably linked to the chemistry, biology and medicine of antiinflammatory glucocorticoids. Sixty years after the first chemical synthesis of cortisone, corticosteroids remain among the top ten most commonly used prescription and over the counter drugs. Basic and clinical studies of glucocorticoid biosynthesis, metabolism and action have trail blazed developments in endocrinology ever since. This article surveys the extraordinary cortisone timeline, from first synthesis until now. The concluding scientific message is that intracrine metabolism of cortisone to cortisol via 11beta hydroxysteroid dehydrogenase type 1 likely sustains local amplification of glucocorticoid action at sites of inflammation throughout the body. The broader message is that the discovery of compound E by Kendall (basic scientist), its large-scale synthesis by Sarett (industrial chemist) and its therapeutic application by Hench (rheumatologist) serves as a paradigm for modern translational medicine. It is concluded that endocrinology will remain a force in health and disease if it continues to evolve sans frontieres at the basic/applied/clinical science interface. A challenge for the Society for Endocrinology is to ensure this happens. PMID- 17911392 TI - Glucocorticoid-induced apoptosis in human decidua: a novel role for 11beta hydroxysteroid dehydrogenase in late gestation. AB - Glucocorticoids play a fundamental role in the endocrinology of pregnancy but excess glucocorticoids in utero may lead to abnormalities of fetal growth. Protection against fetal exposure to cortisol is provided by the enzyme 11beta hydroxysteroid dehydrogenase 2 (11beta-HSD2) located in the human placental trophoblast. By contrast, relatively little is known concerning the function of glucocorticoid-activating 11beta-HSD1, which is strongly expressed within human maternal decidua. To address this we have assessed: i) changes in decidual 11beta HSD1 expression across gestation and ii) the functional role of glucocorticoids in decidua. Human decidua was collected from women undergoing surgical termination of pregnancy in first (n = 32) and second (n = 10) trimesters, and elective caesarean sections in the third trimester (n = 9). Analysis of mRNA for 11beta-HSD1 by real-time RT-PCR showed increased expression in second (9.3-fold, P < 0.01) and third (210-fold, P < 0.001) trimesters. Studies using primary cultures of decidual cells also revealed higher levels of cortisol generation in the third trimester. Changes in decidual 11beta-HSD1 with gestation were paralleled by increased expression of the apoptosis markers caspase-3 and annexin V, particularly in cluster designation (CD)10(-VE) non-stromal cells (20-fold in third trimester relative to first trimester). Apoptosis was also readily induced in primary cultures of third trimester decidual cells when treated with cortisol, cortisone, or dexamethasone (all 100 nM for 24 h). The effect of cortisone but not cortisol or dexamethasone was blocked by an 11beta-HSD inhibitor confirming the functional significance of endogenous cortisol generation. These data show that autocrine metabolism of glucocorticoids is an important facet of the feto placental unit in late gestation and we propose that a possible effect of this is to stimulate programmed cell death in human decidua. PMID- 17911393 TI - Signal transducer and activator of transcription 3-regulated sarcoendoplasmic reticulum Ca2+-ATPase 2 expression by prolactin and glucocorticoids is involved in the adaptation of insulin secretory response during the peripartum period. AB - During pregnancy, the maternal endocrine pancreas undergoes, as a consequence of placental lactogens and prolactin (PRL) action, functional changes that are characterized by increased glucose-induced insulin secretion. After delivery, the maternal endocrine pancreas rapidly returns to non-pregnant state, which is mainly attributed to the increased serum levels of glucocorticoids (GCs). Although GCs are known to decrease insulin secretion and counteract PRL action, the mechanisms for these effects are poorly understood. We have previously demonstrated that signal transducer and activator of transcription 3 (STAT3) is increased in islets treated with PRL. In the present study, we show that STAT3 expression and serine phosphorylation are increased in pancreatic islets at the end of pregnancy (P19). STAT3 serine phosphorylation rapidly returned to basal levels 3 days after delivery (L3). The expression of the sarcoendoplasmic reticulum Ca(2+)-ATPase 2 (SERCA2), a crucial protein involved in the regulation of calcium handling in beta-cells, was also increased in P19, returning to basal levels at L3. PRL increased SERCA2 and STAT3 expressions and STAT3 serine phosphorylation in RINm5F cells. The upregulation of SERCA2 by PRL was abolished after STAT3 knockdown. Moreover, PRL-induced STAT3 serine phosphorylation and SERCA2 expression were inhibited by dexamethasone (DEX). Insulin secretion from islets of P19 rats pre-incubated with thapsigargin and L3 rats showed a dramatic suppression of first phase of insulin release. The present results indicate that PRL regulates SERCA2 expression by a STAT3-dependent mechanism. PRL effect is counteracted by DEX and might contribute to the adaptation of maternal endocrine pancreas during the peripartum period. PMID- 17911394 TI - Cross-regulation of cortisol secretion by adrenocorticotropin and platelet activating factor in perfused guinea pig adrenals. AB - We examined the cross-regulation of signaling between ACTH-and platelet activating factor (PAF)-mediated steroidogenesis in the perfused guinea pig adrenal gland. Our method of in situ perfusion using an artificial medium can evaluate whether cortisol secretion in response to ACTH and PAF is interactive. Treating adrenal glands with 100 pg/ml ACTH diminished the subsequent cortisol response to 10 nM PAF. By contrast, PAF resulted in subsequent potentiation of ACTH-induced cortisol secretion. A mixture of 50 microM L-alpha-1-oleoyl-2-acetyl sn-glycerol (OAG), an activator of protein kinase C (PKC), and 3.3 microM calcium ionophore (A23187), or 10 microM forskolin (FRK) diminished the cortisol response to PAF, whereas that to ACTH was unaffected. Each of PAF, ACTH, or FRK eliminated the cortisol response to OAG plus A23187, whereas that to FRK was unaffected. These data show that the protein kinase A (PKA)-dependent processes activated by ACTH or FRK can interfere with PAF-induced signal transduction at receptor and post-receptor levels. In contrast, PKC-dependent processes activated by PAF promoted ACTH-signaling at receptor and post-receptor level. Cross-regulation between processes activated by PAF receptor-PKC and by ACTH receptor-PKA might function in the multifactorial regulation of adrenocortical steroidogenesis. PMID- 17911395 TI - G-protein-coupled receptors in aldosterone-producing adenomas: a potential cause of hyperaldosteronism. AB - The source of aldosterone in 30-40% of patients with primary hyperaldosteronism (PA) is unilateral aldosterone-producing adenoma (APA). The mechanisms causing elevated aldosterone production in APA are unknown. Herein, we examined the expression of G-protein-coupled receptors (GPCRs) in APA and demonstrated that when compared with normal adrenals, there is a general elevation of certain GPCR in many APA and/or ectopic expression of GPCR in others. RNA samples from normal adrenals (n = 5), APAs (n = 10), and cortisol-producing adenomas (CPAs; n = 13) were used on 15 genomic expression arrays, each of which included 223 GPCR transcripts presented in at least 1 out of 15 of the independent microarrays. The array results were confirmed using real-time RT-PCR (qPCR). Four GPCR transcripts exhibited a statistically significant increase that was greater than threefold when compared with normal adrenals, suggesting a general increase in expression when compared with normal adrenal glands. Four GPCR transcripts exhibited a > 15 fold increase of expression in one or more of the APA samples when compared with normal adrenals. qPCR analysis confirmed array data and found the receptors with the highest fold increase in APA expression to be LH receptor, serotonin receptor 4, GnRH receptor, glutamate receptor metabotropic 3, endothelin receptor type B like protein, and ACTH receptor. There are also sporadic increased expressions of these genes in the CPAs. Together, these findings suggest a potential role of altered GPCR expression in many cases of PA and provide candidate GPCR for further study. PMID- 17911396 TI - Effect of nutrition on the GH responsiveness of liver and adipose tissue in dairy cows. AB - Dairy cows enter a period of energy insufficiency after parturition. In liver, this energy deficit leads to reduced expression of the liver-specific GH receptor transcript (GHR1A) and decreased GHR abundance. As a consequence, hepatic processes stimulated by GH, such as IGF-I production, are reduced. In contrast, adipose tissue has been assumed to remain fully GH responsive in early lactation. To determine whether energy insufficiency causes contrasting changes in the GH responsiveness of liver and adipose tissue, six lactating dairy cows were treated for 4 days with saline or bovine GH when adequately fed (AF, 120% of total energy requirement) or underfed (UF, 30% of maintenance energy requirement). AF cows mounted robust GH responses in liver (plasma IGF-I and IGF-I mRNA) and adipose tissue (epinephrine-stimulated release of non-esterified fatty acids in plasma, IGF-I mRNA, and p85 regulatory subunit of phosphatidylinositol 3-kinase mRNA). Reductions of these responses were seen in the liver and adipose tissue of UF cows and were associated with decreased GHR abundance. Reduced GHR abundance occurred without corresponding reductions of GHR1A transcripts in liver or total GHR transcripts in adipose tissue. In contrast, undernutrition did not alter the abundance of proteins involved in the early post-receptor signaling steps. Thus, a feed restriction reproducing the energy deficit of early lactation depresses GH actions not only in liver but also in adipose tissue. It remains unknown whether a similar reduction of GH action occurs in the adipose tissue of early lactating dairy cows. PMID- 17911397 TI - Ontogenic and nutritional regulation of steroid receptor and IGF-I transcript abundance in the prepubertal heifer mammary gland. AB - In prepubertal cattle, mammary development is characterized by the growth of an epithelial-rich parenchyma (PAR) into the mammary fat pad (MFP). This proliferation and accumulation of mammary epithelial cells require estrogen. Paradoxically, both epithelial cell proliferation and PAR accumulation rate decline with rising plasma estrogen as puberty approaches. The possibility that variation in abundance of estrogen receptors (ERs) in PAR or MFP accounts for a portion of these effects has not been examined in cattle. Additionally, we recently demonstrated that MFP is highly responsive to exogenous estrogen, suggesting that this tissue may play a role in coordinating estrogen's effects on PAR; however, the developing bovine MFP has yet to be studied in detail. To address these hypotheses, Holstein heifers were assigned to planes of nutrition supporting body growth rates of 950 (E) or 650 (R) g/day and harvested every 50 kg from 100 to 350 kg body weight (BW). Post-harvest, their mammary glands were dissected into PAR and MFP compartments. Transcript abundance of genes encoding members of the ER family (ERalpha, ERbeta, and estrogen-related receptor alpha-1 (ERRalpha)) and estrogen-responsive genes (IGF-I and progesterone receptor (PR)) were measured in both mammary compartments by quantitative real-time RT-PCR. Significant expression was detected for all genes in both compartments, with the exception of the ERbeta gene. Transcript abundance of both ERalpha and IGF-I decreased linearly with increasing BW within both compartments. ERRalpha and PR expressions decreased with increasing BW in PAR but not in MFP. Nutrition stimulated ERalpha and ERRalpha expression in the PAR but had no effect on IGF-I or PR in either PAR or MFP. Overall, ERalpha and IGF-I transcript abundance are consistent with the drop in mammary epithelial cell proliferation and PAR accretion observed over development, but do not support a negative effect of nutrition on PAR growth. PMID- 17911398 TI - Relationships of circulating pregnanolone isomers and their polar conjugates to the status of sex, menstrual cycle, and pregnancy. AB - Pregnanolone isomers (PIs) and their polar conjugates (PICs) modulate ionotropic receptors such as gamma-aminobutyric acid or pregnane X receptors. Besides, brain synthesis, PI penetrates the blood-brain barrier. We evaluated the physiological importance of PI respecting the status of sex, menstrual cycle, and pregnancy. Accordingly, circulating levels of allopregnanolone (P3alpha 5alpha ), isopregnanolone (P3beta 5alpha ), pregnanolone (P3alpha 5beta ), epipregnanolone (P3beta 5beta ), their polar conjugates, and related steroids were measured in 15 men (M), 15 women in the follicular phase (F), 16 women in the luteal phase (L), and 30 women in the 36th week of gestation (P) using GC-MS. The steroid levels were similar in M and F, increased about thrice in L and escalated in P (38-410 times compared with F). The PICs were prevalent over the PIs (16-150 times). Higher ratios of 5alpha-PIC to 5alpha-PI found in P indicate the more intensive conjugation of 5alpha-PI during pregnancy. This mechanism probably provides for the elimination of neuroinhibitory P3alpha 5alpha in the maternal compartment. Additionally, our result points to a limited sulfation capacity for neuroinhibitory P3alpha 5beta in P. In contrast to the situation in M, F, and L where the P3alpha 5beta C is the most abundant PIC, and P3alpha 5beta is present in minor quantities compared with the P3alpha 5alpha, P3alpha 5beta may acquire physiological importance during pregnancy, contributing to the sustaining thereof. On the other hand, the declining formation of P3alpha 5beta may participate in the initiation of parturition, given the relative abundance of the steroid, its potency to suppress the activity of oxytocin-producing cells and its effectiveness in uterine relaxation. PMID- 17911399 TI - Fetal programming of colon cancer in adult rats: correlations with altered neonatal growth trajectory, circulating IGF-I and IGF binding proteins, and testosterone. AB - We examined effects of dietary soy protein isolate (SPI) or genistein (GEN; soy isoflavone) during pregnancy on development of colon cancer in male progeny Sprague-Dawley rats. Four groups of rats were used: a lifetime casein-fed group (CAS; control diet), a lifetime SPI-fed group (positive control for protective effect of diet on colon carcinogenesis), a group whose dams received SPI only during pregnancy and CAS thereafter (SPI/CAS), and a group whose dams received CAS+GEN only during pregnancy and CAS thereafter (GEN/CAS). At 47 and 55 days of age, male progeny were administered the intestinal carcinogen azoxymethane (AOM). Tumors, endocrine status, and colon gene expression were evaluated at 20 week post-AOM. The SPI group had 47% decreased colon tumor incidence compared with the CAS group (P<0.05), whereas SPI/CAS, GEN/CAS, and CAS groups did not differ in this regard. Maternal-only SPI increased the percentage of animals bearing multiple colon tumors (P<0.05), an effect not mimicked by GEN. Serum insulin and leptin concentrations were decreased by lifetime SPI (P<0.05), whereas serum IGF I was elevated in the SPI/CAS group (P<0.05). The SPI/CAS group had reduced serum testosterone levels (P<0.05) and exhibited a tendency for increased mucosal expression of IGF-I receptor and glucose transporter-1 mRNAs. Results indicate an effect of dietary protein type during pregnancy on colon tumor multiplicity and colon tissue gene expression, and serum IGF-I and testosterone in progeny rats as later adults. PMID- 17911400 TI - Benign prostatic hyperplasia (BPH) epithelial cell line BPH-1 induces aromatase expression in prostatic stromal cells via prostaglandin E2. AB - Estradiol (E2) level in stroma of benign prostatic hyperplasia (BPH) increases with age, and this increase was associated with an elevated expression of aromatase in prostatic stromal cells (PrSCs). Here, we showed that conditioned medium (CM) of BPH-1 (a benign hyperplastic prostatic epithelial cell line), but not of prostate cancer cell lines (LNCaP, DU-145, and PC-3), stimulates aromatase expression in PrSCs. Cyclooxygenase-2 (COX-2) mRNA level in BPH-1, as well as prostaglandin E2 (PGE2) concentration in BPH-1 CM, was significantly higher than that of prostate cancer cell lines. CM of BPH-1 treated with NS-398 (a specific inhibitor of COX-2) failed to stimulate aromatase expression in PrSCs. And PGE2 can stimulate aromatase expression in PrSCs. Our data suggested that BPH-1 induced aromatase expression in PrSCs through the production of PGE2 in a paracrine mechanism. PMID- 17911401 TI - Activin receptor-like kinase-2 inhibits activin signaling by blocking the binding of activin to its type II receptor. AB - Activin receptor-like kinase-2 (Alk2) has been shown to be a promiscuous type I receptor for the transforming growth factor beta (TGFbeta) family of growth and differentiation factors, such as activin, bone morphogenetic proteins, and Mullerian inhibiting substance (MIS). We have studied the putative role of Alk2 in activin signaling using MA-10 cells, a mouse transformed Leydig cell line, in which endogenous expression of cytochrome P450 c17 hydroxylase/C17-20 lyase mRNA is inhibited by both MIS and activin A. Overexpression of Alk2 in MA-10 cells inhibited the activation of the activin-responsive CAGA-luciferase reporter and, conversely, transfection of siRNA for Alk2 increased the response. In contrast, overexpression of the MIS type II receptor in MA-10 cells increased the activin mediated induction of CAGA-luciferase approximately fivefold, which we hypothesized occurs by MIS type II receptor sequestering endogenous Alk2. Binding experiments with (125)I-labeled activin show that the underlying mechanism of Alk2-mediated inhibition of activin signaling involves Alk2 blocking the access of activin to its type II receptor, which we show can bind Alk2 in the absence of ligand. These results show that the complement of other type I receptors in addition to the ligand-specific type I receptor can provide an important mechanism for modulating cell-specific responses to members of the TGFbeta family. PMID- 17911402 TI - Hydrogen sulphide reduces insulin secretion from HIT-T15 cells by a KATP channel dependent pathway. AB - Hydrogen sulphide (H(2)S), a naturally occurring gas exerts physiological effects by opening K(ATP) channels. Anti-diabetic drugs (e.g. glibenclamide) block K(ATP) channels and abrogate H(2)S-mediated physiological responses which suggest that H(2)S may also regulate insulin secretion by pancreatic beta-cells. To investigate this hypothesis, insulin-secreting (HIT-T15) cells were exposed to NaHS (100 microM) and the K(ATP) channel-driven pathway of insulin secretion was tracked with various fluorescent probes. The concentration of insulin released from HIT-T15 cells decreased significantly after NaHS exposure and this effect was reversed by the addition of glibenclamide (10 microM). Cell viability and intracellular ATP and glutathione (GSH) levels remained unchanged, suggesting that changes in insulin secretion were not ATP linked or redox dependent. Through fluorescence imaging studies, it was found that K(+) efflux occurs in cells exposed to NaHS. The hyperpolarised cell membrane, a result of K(+) leaving the cell, prevents the opening of voltage-gated Ca(2+) channels. This subsequently prevents Ca(2+) influx and the release of insulin from HIT-T15 cells. This data suggest that H(2)S reduces insulin secretion by a K(ATP) channel-dependent pathway in HIT-T15 cells. This study reports the molecular mechanism by which H(2)S reduces insulin secretion and provides further insight into a recent observation of increased pancreatic H(2)S production in streptozotocin-diabetic rats. PMID- 17911403 TI - Pro-inflammatory cytokines increase glucose, alanine and triacylglycerol utilization but inhibit insulin secretion in a clonal pancreatic beta-cell line. AB - We have investigated the effects of prolonged exposure (24 h) to pro-inflammatory cytokines on beta-cell metabolism and insulin secretion using clonal BRIN-BD11 beta cells. Addition of IL-1beta, tumour necrosis factor-alpha and IFN-gamma (at concentrations that did not induce apoptosis) inhibited chronic (24 h) and acute stimulated levels of insulin release (by 59 and 93% respectively), increased cellular glucose and alanine consumption, and also elevated lactate and glutamate release. However, ATP levels and cellular triacylglycerol were decreased while glutathione was increased. We conclude that sub-lethal concentrations of pro inflammatory cytokines appear to shift beta-cell metabolism away from a key role in energy generation and stimulus-secretion coupling and towards a catabolic state which may be related to cell defence. PMID- 17911404 TI - Fibroblast growth factor-23 regulates parathyroid hormone and 1alpha-hydroxylase expression in cultured bovine parathyroid cells. AB - Fibroblast growth factor-23 (FGF23) is a circulating factor that decreases serum levels of inorganic phosphate (Pi) as well as 1,25-dihydroxyvitamin D(3). Recent studies also suggest a correlation between serum levels of FGF23 and parathyroid hormone (PTH) in patients with chronic kidney disease. It is, however, unknown whether FGF23 directly modulates PTH expression, or whether the correlation is secondary to abnormalities in Pi and vitamin D metabolism. The objective of the current study was therefore to elucidate possible direct effects of FGF23 on bovine parathyroid cells in vitro. Treatment of parathyroid cells with a stabilized form of recombinant FGF23 (FGF23(R176Q)) induced a rise in early response gene-1 mRNA transcripts, a marker of FGF23 signaling. FGF23(R176Q) potently and dose-dependently decreased the PTH mRNA level within 12 h. In agreement, FGF23(R176Q) also decreased PTH secretion into conditioned media. In contrast, FGF23(R176Q) dose-dependently increased 1alpha-hydroxylase expression within 3 h. FGF23 (R176Q) did not affect cell viability nor induce apoptosis, whereas a small but significant increase in cell proliferation was found. We conclude that FGF23 is a negative regulator of PTH mRNA expression and secretion in vitro. Our data suggest that FGF23 may be a physiologically relevant regulator of PTH. This defines a novel function of FGF23 in addition to the previously established roles in controlling vitamin D and Pi metabolism. PMID- 17911405 TI - Non-esterified fatty acids and human lymphocyte death: a mechanism that involves calcium release and oxidative stress. AB - Although previous studies have shown that a mixture of fatty acids in similar proportion to that found in human plasma triggers apoptosis of peripheral blood lymphocytes from healthy subjects, the mechanism involved remains unknown. In the present study, we examined whether the effect of a mixture of fatty acids upon human lymphocyte death involves cytochrome c release from the mitochondria, activation of caspases 3, 6, 8, and 9, production of superoxide anion, nitric oxide (NO), increase in cytosolic Ca(2+) levels, and expression of the anti apoptotic 14-3-3 and the pro-apoptotic FasL, bad, and bid proteins. Peripheral blood lymphocytes from healthy subjects were isolated and treated for up to 48 h with increasing concentrations (0.1-0.4 mM) of the fatty acid mixture. Cells were then harvested and thecytochromec release from mitochondrial intermembrane space into cytosol and expression of anti- and pro-apoptotic proteins were investigated by western blot analysis. Activities of caspases 3, 6, 8, and 9 were determined using spectrofluorometric assays. NO production was monitored using DAF-2-FM probe. Cytosolic free calcium concentration ([Ca(2+)](i)) was determined using the fluorescent probe Fura-2-AM. Superoxide anion was assayed using lucigenin and dihydroethidine assays. Lymphocytes treated for 24 h with the fatty acid mixture presented increased cytochrome c release from mitochondria as compared with control lymphocytes without treatment. Activities of caspases 3, 6, and 9 were increased by 146, 22 and 35% respectively by the treatment with 0.4 mM concentration of the fatty acid mixture for 24 h. The expression of bid protein was significantly increased in lymphocytes by 40% at 0.2 mM and by 80% at 0.4 mM fatty acid concentration, whereas FasL, 14-3-3 and bad proteins were not affected by the treatment. Intracellular calcium concentration was increased in a dose dependent manner after 30 min of fatty acid treatment and addition of BSA (0.2%) abolished this increase. Production of NO and superoxide anion was also increased by the fatty acid mixture and BSA loaded in the culture medium prevented this increase. In conclusion, fatty acids induced apoptosis of human lymphocytes by a mechanism that involved cytochrome c release from mitochondria, activation of the caspase cascade, and increase of bid protein content, superoxide and NO production, and of cytosolic calcium concentration. PMID- 17911406 TI - Methimazole inhibits CXC chemokine ligand 10 secretion in human thyrocytes. AB - CXC chemokine ligand 10 (CXCL10) plays a pivotal role in the self-perpetuation of the inflammatory processes in patients with autoimmune thyroid disease. Treatment with methimazole (MMI) reduces serum CXCL10 in patients with Graves' disease. In isolated human thyrocytes, tumor necrosis factor (TNF)alpha demonstrates a potent synergistic effect on interferon (IFN)gamma-induced CXCL10 secretion. We investigated the mechanism underlying the synergism between IFNgamma and TNFalpha and the effect of MMI on CXCL10 secretion in human thyrocytes. A peroxisome proliferator-activated receptor gamma agonist, rosiglitazone (RGZ), a known inhibitor of T helper 1 (Th1)-mediated responses, was also studied for comparison. Experiments were carried out in human thyrocytes isolated from internodular parenchyma of thyroid tissues derived from patients who had undergone surgery for multinodular goiter. ELISA was used to measure CXCL10 levels in culture supernatant. Flow cytometry was used to assess IFNgamma membrane receptor expression. Specific mRNA analysis was performed by Taqman real time PCR. Immunofluorescence was performed to detect nuclear translocation of nuclear factor-kappaB (NF-kappaB). In human thyrocytes, the synergistic effect of TNFalpha with IFNgamma on CXCL10 secretion is due to the upregulation of IFNgamma receptor expression. MMI decreased cytokine-induced CXCL10 secretion by reducing TNFalpha-induced upregulation of the IFNgamma receptor. RGZ decreased the cytokine-induced CXCL10 secretion by impairing NF-kappaB translocation, without affecting IFNgamma receptor. MMI and RGZ targeted thyrocytes with the same pharmacological potency, likely acting throughout different mechanisms. Targeting T helper 1-mediated autoimmune thyroid disease with drugs that impair different intracellular pathways could be a novel pharmacological tool. PMID- 17911407 TI - Ciliary neurotrophic factor promotes survival of neonatal rat islets via the BCL 2 anti-apoptotic pathway. AB - Ciliary neurotrophic factor (CNTF) belongs to the cytokine family and increases neuron differentiation and/or survival. Pancreatic islets are richly innervated and express receptors for nerve growth factors (NGFs) and may undergo neurotypic responses. CNTF is found in pancreatic islets and exerts paracrine effects in neighboring cells. The aim of this study was to investigate possible effects of CNTF on neonatal rat pancreatic islet differentiation and/or survival. For this purpose, we isolated pancreatic islets from neonatal rats (1-2 days old) by the collagenase method and cultured for 3 days in RPMI medium with (CNTF) or without (CTL) 1 nM CNTF. Thereafter, glucose-stimulated insulin secretion (RIA), general metabolism by (NAD(P)H production; MTS), glucose metabolism ((14)CO(2) production), gene (RT-PCR), protein expression (western blotting), caspase-3 activity (Asp-Glu-Val-Asp (DEVD)), and apoptosis (DNA fragmentation) were analyzed. Our results showed that CNTF-treated islets demonstrated reduced glucose-induced insulin secretion. CNTF treatment did not affect glucose metabolism, as well as the expression of mRNAs and proteins that are crucial for the secretory process. Conversely, CNTF significantly increased mRNA and protein levels related to cell survival, such as Cx36, PAX4, and BCL-2, reduced caspase-3 activity, and islet cells apoptosis, suggesting that CNTF does not affect islet cell differentiation and, instead, acts as a survival factor reducing apoptosis by increasing the expression of the anti-apoptotic BCL-2 protein and decreasing caspase-3 activity. PMID- 17911408 TI - Recurrence of the p.R277X/p.R1511X compound heterozygous mutation in the thyroglobulin gene in unrelated families with congenital goiter and hypothyroidism: haplotype analysis using intragenic thyroglobulin polymorphisms. AB - Thyroglobulin (TG) functions as the matrix for thyroid hormone synthesis. Thirty five different loss-of-function mutations in the TG gene have been reported. These mutations are transmitted in an autosomal recessive mode. The objective of this study is to analyze the recurrence of the p.R277X/p.R1511X compound heterozygous mutation in the TG gene in two unrelated families (one Argentinian and another Brazilian) with congenital hypothyroidism, goiter and impairment of TG synthesis. The first and last exon of the TG gene, the exons where previously mutations and single nucleotide polymorphisms (SNPs) were detected, as well as the TG promoter, were analyzed by automatic sequencing in one affected member of the each family. Four microsatellite markers localized in introns 10, 27, 29 and 30 of the TG gene, one insertion/deletion intragenic polymorphism and 15 exonic SNPs were used for haplotype analysis. A p.R277X/p.R1511 compound heterozygous mutation in the TG gene was found in two members of an Argentinian family. The same mutations had been also reported previously in two members of a Brazilian family. We constructed mutation-associated haplotypes by genotyping members of the two families. Our results suggest that the cosegregating haplotype is different in each one of these families. Different haplotypes segregated with the p.R277X and p.R1511 mutations demonstrating the absence of a founder effect for these mutations between Argentinian and Brazilian populations. However, haplotyping of Argentinian patients showed the possibility that the p.R277X alleles might be derived from a common ancestral chromosome. PMID- 17911409 TI - Antibody-induced modulation of TSH receptor post-translational processing. AB - The post-translational processing of the TSH receptor (TSHR) includes intra molecular cleavage with the loss of a 50 amino acid ectodomain region and the formation of two subunits (alpha and beta), followed by likely alpha subunit shedding. TSHR antibodies (TSHR-Abs), which are directed at the ectodomain, may influence thyroid function by stimulating or inhibiting TSHR signaling or may bind without any such influence (the neutral group of antibodies). When we examined the characteristics of a series of monoclonal TSHR-Abs, we found that many were able to inhibit receptor cleavage and enhance TSHR expression. This was especially apparent with the neutral type of TSHR-Abs directed to the cleaved region of the ectodomain (aa 316-366). Indeed, such inhibition appeared to be epitope dependent with TSHR-Abs directed to regions after residues 335-354 showing no such activity. We propose that this aberrant process, whereby TSHR-Abs influence antigen processing, is a novel mechanism for the maintenance and exacerbation of autoimmune thyroid disease. PMID- 17911410 TI - Chromosomes and expression in human testicular germ-cell tumors: insight into their cell of origin and pathogenesis. AB - Human germ-cell tumors (GCTs) are a heterogeneous group of neoplasms. Based on epidemiology, anatomical site of presentation, histology, chromosomal constitution, and pattern of genomic imprinting, GCTs are classified into five entities. Within the testis, three types of GCTs can be diagnosed: type I (teratomas and yolk-sac tumors of neonates and infants); type II (seminomas and nonseminomas); type III (spermatocytic seminomas). Here the focus is on the type II GCTs, the most frequent type in the adult testis (so-called TGCTs). They can also be diagnosed in dysgenetic gonads (an incomplete or defective formation of the gonad, caused by a disturbed process of migration of the germ cells and/or their correct organization in their fetal gonadal ridge), the anterior mediastinum, and pineal/suprasellar region. In the testis, they originate from the malignant counterpart of primordial germ cells/gonocytes, referred to as carcinoma in situ (CIS)/intratubular germ-cell neoplasia unclassified (ITGCNU). CIS/ITGCNU and seminomatous cells are characterized by expression of OCT3/4 and NANOG, while in addition embryonal carcinoma expresses SOX2, all identified as transcription factors related to pluripotency in embryonic stem (ES) cells. With the exception of teratomas, most histological elements of TGCTs are sensitive for (cisplatin-based) chemotherapy; CIS/ITGCNU and seminoma cells are also sensitive to DNA damage induced by irradiation. Similar observations have been made for ES cells and their derivates. Moreover, the genetic constitution of TGCTs (low incidence of mutations and frequent uniparental disomies) can also be linked to characteristics of ES cells, likely related to their specific inability to repair DNA damage and their high sensitivity to apoptotic cell death. The unusual presence of wild-type P53 in TGCTs is explained by specific expression of a cluster of micro-RNAs (miRNAs), that is, hsa-miR 371-373, also expressed in ES cells, which prevents P53-driven cellular senescence upon oncogenic stress. Many characteristics of human TGCTs reflect the nonmalignant counterparts from which they originate. Demonstration of these characteristics, in combination with the knowledge of the abnormal niche of these cells, normally occupied by spermatogonia, allows an informative method for (early) diagnosis. The conclusion is that TGCTs are embryonic cancers found in adults. PMID- 17911411 TI - ETV5 is required for continuous spermatogenesis in adult mice and may mediate blood testes barrier function and testicular immune privilege. AB - The transcription factor Ets-variant gene 5 (ETV5) is essential for spermatogonial stem cell (SSC) self-renewal, as the targeted deletion of the Etv5 gene in mice (Etv5(-/-)) results in only the first wave of spermatogenesis. Reciprocal transplants of neonatal germ cells from wild-type (WT) and Etv5(-/-) testes were performed to determine the role of ETV5 in Sertoli cells and germ cells. ETV5 appears to be needed in both cell types for normal spermatogenesis. In addition, Etv5(-/-) recipients displayed increased interstitial inflammation and tubular involution after transplantation. Preliminary studies suggest that the blood-testis barrier (Sertoli-Sertoli tight junctional complex) is abnormal in the Etv5(-/-) mouse. PMID- 17911412 TI - Toward understanding the core meiotic transcriptome in mammals and its implications for somatic cancer. AB - Progression through meiotic development is in part controlled by an expression program that coordinates the timing of induction and time of function of numerous loci essential for the process. Whole-genome profiling of male germline expression in mouse, rat, and human provides important clues about the transcriptional regulatory machinery that drives the expression of its target genes. Among several thousand genes differentially expressed between testicular Sertoli and germ cells, a subset of conserved loci display highly similar meiotic and postmeiotic profiles across rodents and Homo sapiens. Mouse genes transcribed in the germline, but not in somatic control tissues, are frequently found to be important for sexual reproduction, thus correlating potentially specific expression and essential function in the male germline. In silico promoter analysis yields insight into DNA sequence conservation and the distribution of known regulatory elements within potential promoter regions of meiotic and postmeiotic genes. Some genes strongly expressed in male gonads are implicated in cancer, thus supporting the idea that gametogenesis and tumorigenesis may share molecular functions. Transcriptome, proteome, and protein network data reveal the kinetics of mRNA synthesis and translation in the germline, and help identify novel potentially important genes previously unassociated with the mammalian male germline. PMID- 17911413 TI - Proceedings of the 5th International Congress on Autoimmunity, November 29 December 3, 2006, Sorrento, Italy. Part B. PMID- 17911414 TI - Recent advances in the genetics of autoimmune diseases. AB - Autoimmune diseases in general are complex genetic diseases where genes and environment interact in unknown ways. In recent years technologies have advanced our knowledge and new genes are being identified very rapidly. We can expect that most genes of major importance for the various autoimmune diseases will be discovered in the coming 5 years. The real challenge comes when we try to understand the mechanisms through which these genes confer disease susceptibility and how the interaction with environment takes place such that clinical expression of the disease results. PMID- 17911415 TI - Immunoglobulin-like transcripts as risk genes for autoimmunity. AB - Risk genes for multiple sclerosis (MS) are localized in the gene regions 6p21-11 and 19q13, the latter harboring the genes of the immunoglobulin-like transcripts (ILTs). ILTs are a family of activating and inhibitory receptors expressed on antigen-presenting cells (APCs) as well as on natural killer (NK) and T cells. Because of the inhibitory function of ILT2 and ILT4 and their binding to human leukocyte antigen (HLA)-G, they play a role in immune tolerance and may be important in pathogenesis of autoimmunity. ILT6 shows presence-absence variability and is produced by macrophages in a soluble form. ILT6 deletion is associated with MS. Furthermore, ILT6 activates T cell proliferation and is therefore a candidate gene for autoimmune disorders. PMID- 17911416 TI - Genetic polymorphisms of transforming growth factor beta-1 promoter and primary biliary cirrhosis in Japanese patients. AB - As suggested by concordance rates in twins, genetic factors are critical to the susceptibility and progression of primary biliary cirrhosis (PBC). Among cytokines, transforming growth factor beta-1 (TGF-beta1) plays an important role in autoimmunity and liver fibrosis and a TGF-beta1 receptor knockout mouse has been recently proposed as a model for PBC. The promoter region of the TGF-beta1 gene has two single nucleotide polymorphisms (SNPs) at positions -800 and -509, which influence serum concentrations of latent and active TGF-beta1. We studied genomic DNA from 65 Japanese patients with PBC and 71 matched healthy controls for the association of TGF-beta1 SNPs analyzed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) with susceptibility and disease progression of PBC. The -800 G to A SNP was not found in the Japanese population and no significant difference in the distribution of TGF-beta1 promoter gene -509 SNP was found between PBC cases and controls. Further, TGF beta1 genotypes failed to correlate with clinical parameters, including histological stage and prognostic score. In conclusion, the TGF-beta1 promoter gene SNPs are not associated with disease susceptibility or progression in Japanese patients with PBC. PMID- 17911417 TI - Genetic background of anticyclic citrullinated peptide autoantibody production in Hungarian patients with rheumatoid arthritis. AB - Polymorphisms of the peptidylarginine deiminase 4 (PADI4) gene encoding for the isoenzyme that converts arginyl into citrullyl residues have been shown to contribute to susceptibility to rheumatoid arthritis (RA), depending on the population studied. We aimed at determining whether PADI4 single nucleotide polymorphisms (SNPs) are associated with RA in a Hungarian population. The relationship between anticyclic citrullinated peptide (anti-CCP) production and HLA-DRB1 alleles encoding the shared epitope (SE) was also investigated. DNA samples were obtained from RA (n = 261) patients and from control donors (n = 120). HLA-DRB1 genotyping was carried out by polymerase chain reaction (PCR) with sequence-specific priming. PAD4_92 G/C and PAD4_104 T/C SNPs were genotyped using real-time PCR allele discrimination. Autoantibodies against CCP were detected by ELISA. All healthy controls tested anti-CCP negative, whereas 171 (66%) RA patients were anti-CCP positive. No significant difference in allele or genotype frequencies were found between RA patients and controls for any of the PADI4 SNPs. Anti-CCP seropositivity was unrelated to these two SNPs. No association was found between any of the PADI4 SNPs and HLA-DRB1 subtypes. Presence of the HLA RB1 SE alleles was significantly associated with anti-CCP seropositivity; HLA DRB1*0401 and HLA-DRB1*1001 carriers showed the strongest association. In conclusion, our data suggest that polymorphisms of the PADI4 gene are not associated with rheumatoid arthritis and are unlikely to be responsible for the presence of anti-CCP autoantibodies in a white Hungarian population. HLA-DRB1 SE alleles, however, may significantly contribute to the genetic determination of anti-CCP production in Hungarian patients with RA. PMID- 17911418 TI - Profiling meta-analysis reveals primarily gene coexpression concordance between systemic lupus erythematosus and rheumatoid arthritis. AB - Consensus gene expression profiling by meta-analysis of 4,500 cDNA sequence microarray data obtained from patients with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) was assembled and systematically analyzed. The normalized data were statistically analyzed by the significance analysis of microarray (SAM) program (false discovery rate 200 pmol/L and antiGAD > 50 ng/mL. We aimed to find a possible genetic discrimination among different types of autoimmune diabetes. To accomplish this goal, we analyzed DNA samples from 31 LADA patients, 75 patients with adult-onset type 1 diabetes mellitus, 188 type 1 diabetic children, and 153 healthy adult individuals. We studied five genetic loci on chromosomes 6, 11, 4, and 14: HLA DRB1 and DQB1 alleles, major histocompatibility complex (MHC) class I related gene-A (MIC-A) microsatellite polymorphism, interleukin (IL)-18 single nucleotide polymorphism, the microsatellite polymorphism of nuclear factor kappa B gene (NF-kappaB1), and the single nucleotide polymorphism of a gene for its inhibitor (NF-kappaBIA). HLA-DR3 was detected as the predisposition allele for LADA (OR = 4.94, P < 0.0001). Further we found a statistically significant increase of NF-kappaBIA AA genotype (OR = 2.68, P < 0.01). On the other hand, DRB1*04, which is linked with DQB1*0302, was observed as a risk factor in patients with type 1 diabetes mellitus (T1DM) onset after 35 years of age (OR = 10.47, P < 0.0001 and OR = 9.49, P < 0.0001, respectively). There was also an association with MIC-A5.1 (OR = 2.14, P < 0.01). Statistically significant difference was found in the distribution of IL-18 promoter -607 (C/A) polymorphism between LADA and T1DM in adults (P < 0.01). We conclude that all subgroups of autoimmune diabetes have partly different immunogenetic predisposition. PMID- 17911430 TI - Genetic control of immune response in carriers of the 8.1 ancestral haplotype: correlation with levels of IgG subclasses: its relevance in the pathogenesis of autoimmune diseases. AB - Ancestral haplotype (AH) 8.1(HLA-A1, Cw7, B8, TNFAB*a2b3, TNFN*S, C2*C, Bf*s, C4A*Q0, C4B*1, DRB1*0301, DRB3*0101, DQA1*0501, DQB1*0201) seems to be associated with susceptibility to autoimmune diseases. Different mechanisms are probably involved in increasing autoimmunity, such as unbalanced cytokine production and the lack of C4A protein. So AH 8.1 modifies immune response in many ways. In this study we demonstrate that IgG2 serum levels were significantly lower in 8.1 AH carriers than in 8.1 AH non-carriers. On the contrary, as regards IgG1, IgG3, IgG4 serum levels, no significant differences were observed between the two groups. In AH 8.1 carriers low IgG2 levels might take to slower clearance of the infectious agent and hence to a lasting presence of it. The persistence of infectious antigens could determine an increased production of autoantibodies with a higher risk of cross-reactions. PMID- 17911431 TI - Genetic dissection of autoimmune polyendocrine syndrome type 2: common origin of a spectrum of phenotypes. AB - Autoimmune diseases constitute a heterogeneous group of disorders characterized by the loss of immune tolerance to self-antigens. Despite their distinct clinical picture, there is growing evidence that common molecular mechanisms may contribute to the whole spectrum of autoimmune diseases. This theory is strongly supported by the existence of the autoimmune polyendocrine syndromes (APS). Thus, the clinical diagnosis of APS1 is made in an individual who presents with at least two out of three cardinal symptoms, namely autoimmune Addison's disease, autoimmune hypoparathyroidism, and mucocutaneous candidiasis. APS1 is a rare autosomal recessive syndrome caused by mutations in the autoimmune regulator (AIRE) gene. APS2, which occurs at a much higher frequency, is classically defined as the coexistence of autoimmune Addison's disease, autoimmune thyroid disease, and/or type 1 diabetes. In contrast to APS1, the precise modes of inheritance and the genetic causes underlying APS2 remain unknown. Identification of genetic factors predisposing to this syndrome may contribute to our understanding of common mechanisms involved in autoimmunity. PMID- 17911432 TI - Association of the 5A/6A promoter polymorphism of the MMP-3 gene with the radiographic progression of rheumatoid arthritis. AB - Matrix metalloproteinases (MMPs) as a family of zinc-dependent endopeptidases have been involved in remodeling the extracellular matrix (ECM) in rheumatoid arthritis (RA). In RA patients synovial fluid and serum include enhanced levels of MMP-3. The 5A/6A polymorphism in the MMP-3 gene promoter can contribute to the severity of RA on account of a higher promoter activity of the 5A allele in vitro. The aim of the study was to associate the 5A/6A polymorphism of the MMP-3 gene with radiographic progression of RA. A total of 128 RA patients according to the ACR criteria were available for the study. Radiographs of both hands, obtained from all RA patients, were scored using the modified Sharp/van der Heijde method and the Steinbrocker method. The total Sharp score (TSS) and the annual radiographic progression rate (TSS/year) were calculated. Significant association with the 5A/6A polymorphism was found between patients with TSS/year 1.00 in allelic frequencies (Pa = 0.046) and also in genotype distribution (Pg = 0.04). Compared to other genotypes the prevalence of 5A/5A genotype was lower within patients with TSS/year or=80%) in 20 of 70 GD patients (28.6%) and 11 of 43 HT patients (25.6%), giving a total of 31 of 113 AITD patients (27.4%) with skewed XCI. In contrast, only 5 of 58 healthy controls had skewed XCI (8.6%). Statistical analysis confirmed that XCI skewing was significantly associated with AITD (P = 0.004, OR = 4.0), demonstrating that the degree of XCI is an important contributor to the increased risk of females in developing AITD. PMID- 17911436 TI - Mycophenolate mofetil and atherosclerosis: results of animal and human studies. AB - The immunosuppressive agent mycophenolate mofetil (MMF) has beneficial effects in cardiac transplant patients beyond the suppression of tissue rejection. Patients with regimens containing MMF experience diminished intimal thickening and cardiac allograft vasculopathy compared to patients treated with azathioprine. Studies have shown that diet-induced atherosclerosis (a related vasculopathy) is a chronic inflammatory process, and so MMF has also been used to reduce atherosclerosis in a rabbit model of hyperlipidemia. These data hold out the intriguing possibility that MMF might be a viable primary or secondary preventive agent in people at significant risk for atherosclerosis. PMID- 17911437 TI - T cell receptor gene therapy for autoimmune diseases. AB - The current quality of autoimmune disease treatments is not satisfactory in regard to efficacy and safety. Antigen-specific immunotherapy is a future therapy that could achieve maximal efficacy with minimal adverse effects. T cells are essential components in antigen-specific immunity. However, we do not have a sufficient strategy for manipulating antigen-specific T cells. We propose that T cell receptor (TCR) gene transfer is a hopeful approach for antigen-specific immunotherapy. We confirmed the efficacy of TCR gene therapy in animal models of systemic autoimmune disease and arthritis. In lupus-prone NZB/W F1 mice, nucleosome-specific TCR and CTLA4Ig transduced cells suppressed autoantibody production and nephritis development. In the therapeutic experiment of collagen induced arthritis (CIA), arthritis-related TCRs were isolated from single T cells accumulating in the arthritis site. Arthritis-related TCR and TNFRIg transduced cells or TCR and Foxp3 transduced cells suppressed arthritis progression and bone destruction. Therefore, engineered antigen-specific cells manipulated to express appropriate functional genes could be applied to specific immunotherapy. PMID- 17911439 TI - Efficacy of intravenous immunoglobulin in multifocal motor neuropathy. AB - Multifocal motor neuropathy is a distinct entity, whose treatment differs from that of other chronic immune-mediated neuropathies, mainly chronic inflammatory demyelinating polyradiculoneuropathy, and its variant, multifocal acquired demyelinating sensory and motor neuropathy, although they share some electrophysiological characteristics. From the first descriptions, intravenous immunoglobulins (IVIg) have been considered to be the gold standard of treatment for multifocal motor neuropathy. However, if the effectiveness of IVIg has been confirmed by several randomized, double-blind, placebo-controlled trials, only a few patients experience persistent improvement after a single or few courses of therapy, and the long-term efficacy of IVIg in this disease is currently debated. Consequently, there is a need for new therapeutic strategies that focus on the effects and the costs of this therapy over long-term follow-up. PMID- 17911438 TI - CD44 involvement in autoimmune inflammations: the lesson to be learned from CD44 targeting by antibody or from knockout mice. AB - CD44 is a multistructural and multifunctional glycoprotein, the diversity of which is generated by alternative splicing. In this communication we review some aspects related to CD44 structure and function in experimental autoimmune inflammation, focusing on research performed in our own laboratory. We have found that CD44 targeting by antibody, passively injected into DBA/1 mice with collagen induced arthritis (CIA) and NOD mice with type I diabetes or actively generated by CD44 cDNA vaccination of SJL/j mice with autoimmune encephalomyelitis, markedly reduced the pathological manifestations of these diseases by attenuating cell migration of the inflammatory cells and/or by their apoptotic killing. However, genetic deletion of CD44 by knockout technology enhanced the development of CIA because of molecular redundancy mediated by RHAMM (a receptor of hyaluronan-mediated motility). The mechanisms that stand behind these findings are discussed. PMID- 17911440 TI - New autoantibodies in pediatric opsoclonus myoclonus syndrome. AB - Opsoclonus-myoclonus syndrome (OMS) is a rare neurologic disorder comprising the main symptoms of eye-movement disturbances, muscle jerks, and severe ataxia. In children and adults, some cases are associated with a tumor as a paraneoplastic syndrome, whereas in children the paraneoplastic form is almost exclusively associated with neuroblastoma. The detection of autoantibodies in some OMS sera led to the hypothesis that the syndrome is of autoimmune origin. Beside autoantibodies against intracellular proteins, such as anti-Hu, alpha-enolase, and KHSRP, specific binding of autoantibodies to the surface of neuroblastoma cells and cerebellar granular neurons have been found. Antiproliferative and proapoptotic effects of these autoantibodies on neuroblastoma cell lines were noted as well. These results support the concept of a humoral autoimmune process in the pathogenesis of OMS. PMID- 17911441 TI - Current state of biologicals in the management of systemic vasculitis. AB - Conventional immunosuppressive treatment of systemic vasculitides has improved their often fatal outcome, but is burdened by cytotoxic side effects and frequent relapses. Recent advances in the therapy of systemic vasculitides with biologicals have helped to establish new options for patients resistant to conventional treatment. Moreover, early intervention aiming to interfere with specific targets important in the break of tolerance and/or persistence of the autoimmune response might further improve the prognosis of autoimmune vasculitides such as antineutrophil cytoplasmic autoantibody (ANCA)-associated vasculitides (AAV). In vitro and in vivo studies suggest that the interaction of ANCA and cytokine (TNF-alpha, IL-1)-primed neutrophils results in premature neutrophil activation and degranulation, subsequent endothelial cell damage, and further leukocyte recruitment. For one of the AAV, Wegener's granulomatosis, recent ex vivo data have provided evidence that WG-granulomata might provide the necessary "proinflammatory environment" for the break of tolerance and display features of lymphoid-like tissue neoformation, in which autoimmunity to "Wegener's autoantigen" proteinase 3 PR3 could be sustained. Blocking TNF-alpha and eliminating autoreactive B cells seem promising treatment targets to interfere with these fundamental disease processes. While the recombinant TNF alpha receptor/IgG1 fusion protein etanercept, in addition to standard therapy with subsequent tapering of standard medications, was found to be not effective for maintenance of remission, open clinical studies suggest a beneficial effect of the anti-TNF-alpha antibody infliximab in addition to standard therapy for the induction of remission in patients with refractory AAV. Peripheral B cell depletion with the anti-CD20 antibody rituximab also induced remissions in AAV in uncontrolled trials. PMID- 17911442 TI - Interstitial lung disease associated with systemic sclerosis: what is the evidence for efficacy of cyclophosphamide? AB - Interstitial lung disease (ILD) is a common manifestation of systemic sclerosis (SSc) that may be responsible for severe restrictive lung disease and represents one of the two main causes of disease-related death in SSc patients. Since 1993, the beneficial effect of oral or intravenous cyclophosphamide (CYC) in the treatment of SSc-related ILD has been reported in retrospective studies, one study showing improvement of pulmonary function test scores and/or chest computed tomography at 1 year and improvement of survival at 16 months. The results of two controlled trials were recently reported. The Scleroderma Lung Study, a prospective randomized placebo-controlled trial, included 158 patients of whom 145 completed at least 6 months of treatment. The course of forced vital capacity (primary outcome) adjusted at 1 year was significantly better in the group treated with oral CYC (P < 0.03), although the effect of CYC was minor. The Fibrosing Alveolitis in Scleroderma Trial (FAST) included 45 patients with SSc related ILD who were randomized to receive prednisolone (20 mg per day) and 6 CYC infusions (600 mg/m monthly) or placebo. This trial did not demonstrate significant improvement of the primary or secondary endpoints in the active treatment group versus placebo. Since with the exception of the study of Silver et al. none of the patients included in retrospective or prospective studies were selected on the basis of progression of ILD. Since only a minority of SSc patients develops severe ILD, we propose that further studies evaluating CYC should focus on the subgroup of SSc patients with worsening ILD. PMID- 17911443 TI - Treatment of autoimmune disease with rabbit anti-T lymphocyte globulin: clinical efficacy and potential mechanisms of action. AB - The rabbit anti-T lymphocyte globulins (rATGs) are immune-suppressive anti-T cell agents with beneficial effects in solid organ and hematological transplantation. The present review evaluates the potential mechanisms of rATGs and their impact on pilot and exploratory studies of diffuse cutaneous systemic sclerosis (scleroderma-SSc), inclusion body myositis (IBM), vasculitis, and type 1 diabetes mellitus (T1DM). The rATGs are associated with improvements in well-defined parameters of clinical autoimmunity: insulin usage, tissue inflammation, and systemic organ functions. Meta-analysis of a retrospective database of SSc, N = 196 and two prospective randomized pilot studies; IBM, N = 11 and T1DM, N = 17 shows a two- to ninefold increase in the relative response to treatments with intravenous infusions of rATG. The rATGs deplete T cells and are associated with increases in the percentage of CD25+ T cell subsets. This may underlie the apparent long-lasting immunomodulation associated with these agents. The future optimization of rATG adjunct therapy requires statistically powered-controlled prospective trials of rATG dose-finding and timing of administration. The potential mechanisms of rATGs:depletion of autoreactive T cells, generation CDCD25+Foxp3+ regulatory T cells (Tregs), and the acquisition of regulatory immune cell functions, need to be examined in patients prior to rATG infusion and at time intervals following rATG treatment to identify those mechanisms relevant to the improvement of their clinical outcome. PMID- 17911444 TI - Bromocriptine during pregnancy in systemic lupus erythematosus: a pilot clinical trial. AB - Bromocriptine (BRC) prevents postpartum flare in lupus patients. However, its potential role in protecting lupus pregnancy from maternal-fetal complications has not been studied. The objective of the study was to explore the role of oral BRC during pregnancy in patients with systemic lupus erythematosus (SLE). Pregnant SLE patients were randomized into two groups: group 1 received BRC 2.5 mg/day and prednisone 10 mg/day; group 2 received prednisone 10 mg/day. These treatments were administered from 25 to 35 weeks of gestation. Prolactin (PRL) levels were determined at 25, 30, and 35 weeks. The SLE Pregnancy Disease Activity Index, maternal-fetal outcome including preterm birth, fetal loss, premature rupture of membrane (PRM), low birth weight, and preeclampsia/eclampsia were evaluated. We studied 20 patients (10 in each group). A significant decrease of PRL levels in group 1 compared to group 2 at week 30 and at week 35 was found. No patients in the BRC group had flares and three from group 2 had SLE activity. None of the patients in group 1 had PRM but three patients in group 2 did. Eighty percent of pregnancies ended in birth at term in group 1 and 50% in group 2. There was no fetal loss in both groups. Mean birth weight was higher in group 1 than in group 2 (P < NS). BRC was well tolerated. This is the first clinical trial of BRC in SLE pregnancy. Our pilot study suggests that BRC may play a role in the prevention of maternal-fetal complications, such as PRM, preterm birth, and active disease. PMID- 17911445 TI - Efficacy and safety of intravenous immunoglobulin in patients with metastatic melanoma. AB - We have previously reported studies performed both in vitro and in laboratory animals, as well as a case study in humans, suggesting that intravenous immunoglobulin (IVIG) may be beneficial in the treatment of malignancies, including metastatic melanoma. As part of a phase II open label trial, we have administered IVIG to nine patients with metastatic melanoma who had been heavily treated. In two of nine (22%) patients treated every 3 weeks with IVIG (1 g/kg body weight), the disease stabilized. One patient had stable disease for 8 months; the other for 3 months. No serious adverse events (AEs) attributable to IVIG were observed. We conclude that IVIG therapy may be useful for the treatment of metastatic melanoma. Furthermore, we suggest that the effects of IVIG therapy might be enhanced by its use as an adjuvant in patients without evidence of disease following surgery. PMID- 17911446 TI - Acute ST elevation myocardial infarction during intravenous immunoglobulin infusion. AB - Intravenous immunoglobulin (IVIG) preparations are increasingly being used in the treatment of autoimmune disorders. This treatment is regarded as generally safe, and most of the adverse effects associated with IVIG administration are mild and transient. This paper reports a 72-year-old patient with known ischemic heart disease admitted for a Guillain-Barre syndrome variant, who developed acute ST elevation myocardial infarction (MI) during the first hours of IVIG infusion. The literature on acute MI during IVIG treatment is reviewed. Evaluation of each patient for cardiovascular risk prior to IVIG treatment is recommended as is the assessment of risk versus benefit. If IVIG is prescribed, we propose close monitoring and slow infusion rate. PMID- 17911447 TI - Autoantibody production in anti-TNF-alpha-treated patients. AB - Targeting tumor necrosis factor alpha (TNF-alpha) has offered an additional therapeutic strategy against several rheumatic inflammatory disorders. The current use of TNF-alpha inhibitors allows physicians who manage these diseases and patients themselves to testify to an extraordinary efficacy, even though caution for possible adverse events must be maintained. Among these, the occurrence of autoimmune phenomena, encompassing new autoantibody formation and triggering of clinical manifestations, continues to be noted in published reports. Here, we review the current knowledge regarding the autoimmune phenomena linked to anti-TNF-alpha therapy in patients with rheumatic inflammatory disorders. PMID- 17911448 TI - Anti-IL-17A autovaccination prevents clinical and histological manifestations of experimental autoimmune encephalomyelitis. AB - Excessive or inappropriate production of IL-17A has been reported in diseases such as rheumatoid arthritis, asthma, and multiple sclerosis. The potential clinical relevance of these correlations was suggested by the protective effects of anti-IL-17A monoclonal antibodies in various mouse disease models. However, the chronic nature of the corresponding human afflictions raises great challenges for Ab-based therapies. An alternative to passive Ab therapy is autovaccination. Covalent association of self-cytokines with foreign proteins has been reported to induce the production of antibodies capable of neutralizing the biological activity of the target cytokine. We recently reported that cross-linking of IL 17A to ovalbumin produced highly immunogenic complexes that induced long-lasting IL-17A-neutralizing antibodies. Vaccinated SJL mice were completely protected against experimental autoimmune encephalomyelitis (EAE) induced by proteolipid protein peptide (PLP 139-151), and a monoclonal anti-IL-17A Ab (MM17F3), derived from C57Bl/6 mice vaccinated against IL-17A-OVA, also prevented disease development. Here we report that this Ab also protects C57Bl/6 mice from myelin oligdendrocyte glycoprotein (MOG)-induced EAE. Histological analysis of brain sections of C57Bl/6 mice treated with MM17F3 showed a complete absence of inflammatory infiltrates and evidence for a marked inhibition of chemokine and cytokine messages in the spinal cord. These results further extend the analytical and therapeutic potential of the autovaccine procedure. PMID- 17911449 TI - Human immunoglobulins in intraocular inflammation. AB - Chronic intraocular inflammation (IOI) is a heterogeneous group of rare diseases, which represents one of the leading causes of acquired and treatable blindness in adults. The main anatomical site of inflammation is the uveal tract, which is the vascular organ of the eye, but uveitis is now used to describe IOI more globally. In around 40% of the cases of uveitis an underlying systemic disease, often of autoimmune origin, can be identified. In autoimmune diseases with IOI, uveitis may be the first clinical manifestation and may represent the most severe sign. Studies in animal models, especially in experimental autoimmune uveitis (EAU), offer the opportunity to investigate the pathogenicity of these disorders. The conventional treatment of IOI includes corticosteroids and immunosuppressive agents, which are efficient in around one-half of the patients, but their effectiveness is also limited by their iatrogenicity. The effects of intravenous immunoglobulins (IVIGs) on ocular inflammation have been investigated in a wide spectrum of autoimmune/systemic diseases. Most of the publications are case series or open trials. They show favorable results in a subset of indications, including mainly ocular cicatricial pemphigoid (OCP), Vogt-Koyanagi-Harada (VKH) syndrome, or birdshot disease. Efficacy results are more debated in other conditions such as inflammatory demyelinating optic neuritis. In other diseases with IOI only case reports are available, suggesting that IVIGs may be of some interest. These observations support the need for controlled trials to demonstrate the efficacy of IVIGs and assess their potential steroid-sparing effect. PMID- 17911450 TI - Fermented wheat germ extract (avemar) inhibits adjuvant arthritis. AB - Anti-inflammatory efficacy of the fermented wheat germ extract (FWGE, Avemar) in the rat adjuvant arthritis (AA) model was examined. To Wistar rats with AA, different doses of FWGE and anti-inflammatory drugs (indomethacin, dexamethasone) as monotherapies were administered and FWGE and either diclofenac or dexamethasone were also given in combination. Besides plethysmographies of the paws, histological investigations of synovial tissues were also performed along with detection of CD4+ and CD8+ T lymphocytes. Gene expressions of COX-1 and 2 were determined by real-time polymerase chain reaction (PCR). FWGE monotherapy significantly inhibited the development of the secondary (immune-mediated) response in AA, and dexamethasone and indomethacin exerted inhibitory effects in a degree comparable to that of FWGE. Histological analysis of the affected joints confirmed the results. FWGE inhibited COX-1 and -2, while indomethacin enhanced COX-2 gene expressions. FWGE had an additive interaction with diclofenac. It is concluded that FWGE has significant anti-inflammatory efficacy confirmed by plethysmography, histology, and real-time PCR. PMID- 17911451 TI - Allogenic mixed chimerism induced by nonlymphoablative regimen including donor BMT with low-dose TBI and anti-CD40L cured proliferative glomerulonephritis in lupus mice. AB - Allogeneic mixed chimerism achieved by low-dose total body irradiation (TBI) and anti-CD40L monoclonal antibody (mAb) with donor bone marrow transplantation (BMT) and host T cell depletion overcomes both allo- and autoimmunity. We investigated whether a similar regimen without T cell depletion cured diffuse proliferative glomerulonephritis. Male BXSB mice (H-2b) were injected with 20 x 10(6) BALB/c (H 2d) BM cells. When indicated, 3 Gy TBI on day -1 and anti-CD40LmAb (2 mg) on day 0 of BMT was given. Skin grafting was performed 1 day after BMT. BXSB mice were divided into four groups--I: BMT with TBI and anti-CD40LmAb; II: TBI; III: TBI and anti-CD40LmAb; and IV: no treatment. Chimerism in peripheral blood was analyzed. The kidney was examined histologically. TBI with anti-CD40LmAb and BMT allowed induction of multilineage mixed chimerism and donor-specific tolerance to skin grafts without graft-versus-host disease (GVHD). There was significant decrease in glomerular PAS-positive material deposition score, glomerular cell numbers, IgG, and C3 deposition in chimeric mice. All chimeric mice survived. Allogeneic mixed chimerism induced by a less toxic, nonlymphoablative regimen achieved allograft tolerance and cured glomerulonephritis in BXSB lupus mice. PMID- 17911453 TI - Targeting vascular adhesion protein-1 to treat autoimmune and inflammatory diseases. AB - The development of new, safe, and effective anti-inflammatory drugs represents a major challenge for the pharmaceutical industry, as well as a significant opportunity. The increasing prevalence of chronic inflammatory and autoimmune diseases associated with an aging population has led to an intense effort to discover new anti-inflammatory drug targets and drugs acting against them. This review highlights the recent progress made in developing therapies directed against an endothelial cell adhesion molecule called vascular adhesion protein (VAP)-1 for the treatment of chronic inflammatory disease as well as highlighting other therapeutic opportunities offered by this vascular target. PMID- 17911452 TI - Interleukin-20 as a target in psoriasis treatment. AB - Interleukin-20 (IL-20) is a new member of the IL-10 cytokine family discovered by a structural algorithm. IL-20 transgenic mice displayed skin abnormalities reminiscent of psoriasis, a finding that has prompted the investigation of this new interleukin in relation to this disease. This article reviews the role of IL 20 and its implication in psoriasis. It is shown that IL-20 and its receptors are found in human skin and that IL-20 is involved in proliferation, angiogenesis, and chemotaxis, all characteristics of psoriasis. We demonstrated that IL-20 induced the thickening of human epidermis in vivo; however, this thickening does not seem to be related to a direct effect of IL-20 on hyperproliferation since the growth of normal human epidermal keratinocytes (NHEKs) cultured in vitro was not affected by IL-20. On the other hand, in vitro, IL-20 stimulated human peripheral blood mononuclear cells (PBMCs) to produce proinflammatory cytokines and, in vivo, IL-20 in combination with PBMCs induced psoriasis. This may suggest that IL-20 indirectly exerts its proliferative effects on keratinocytes via immune cells present in the skin. Finally, we found that blocking IL-20 signaling in psoriasis improves psoriasis, suggesting that IL-20 is a potential target in psoriasis treatment. PMID- 17911454 TI - Rapid and deep control of inflammation in rheumatoid arthritis with infliximab and its correlation with acute-phase reactants. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease with predominant joint involvement and possible systemic compromise, which leads to a handicapped status and poor quality of life. An optimal approach to treat RA requires early and intensive intervention with close monitoring of treatment response. Tumor necrosis factor (TNF) blockers are recommended in cases of active RA after the unsuccessful use of effective disease-modifying antirheumatic drugs (DMARDs); even adding them to treatment or replacing these drugs. Anti-TNF therapies have been demonstrated to reduce significant joint damage and to relieve symptoms during a prolonged time (see Scott and Kingsley, 2006). The efficacy of infliximab in an open-label trial is summarized with respect to speed of onset of action, durability of response, and its correlation between clinical and laboratory parameters. Safety for long-term treatment is also summarized. We studied 105 RA patients with more than 3 years' history of disease during 24 months on i.v. infliximab (75 completed study). We evaluated ACR responses at base line, and at 1, 6, 12, 16, 52, 77, and 104 weeks. Morning stiffness, swollen and tender joints, HAQ, SF-36% (PCS/MCS), polymerase chain reaction (PCR), erythrosedimentation rate (ESR), transaminases, rheumatoid factor (RF) levels, hemogram, and adverse events profile were all assessed. The treatment offered rapid and sustained clinical improvements as revealed by ACR responses and marked changes in the parameters previously described. Important changes were made in functional status and acute-phase reactants. Finally, infliximab was considered well tolerated and did not affect the safety profile of this trial. PMID- 17911455 TI - Dual inhibition of dipeptidyl peptidase IV and aminopeptidase N suppresses inflammatory immune responses. AB - The ectopeptidases dipeptidyl peptidase IV (DP IV, CD26) and aminopeptidase N (APN, CD13) are known to regulate T cell activation. Since selective inhibitors of DP IV and APN suppress DNA synthesis and cytokine production of stimulated T cells in a TGF-beta1-dependent manner, we tested whether combined application of DP IV and APN inhibitors enhances this immunomodulatory effect. The results show that simultaneous application of DP IV and APN inhibitors significantly suppressed DNA synthesis in mitogen- or anti-CD3-stimulated human T cells in vitro when compared to the use of a single DP IV or APN inhibitor. Moreover, the combined action of DP IV and APN inhibitors markedly increased TGF-beta1 production associated with the observed immunosuppressive effects. In vivo, targeting both DP IV and APN led to a potent treatment of experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis (MS). This review summarizes the evidence for the role of both enzymes in T cell activation in vitro and in vivo and provides a rationale for using combined and dual peptidase inhibitors to treat autoimmune diseases like MS. PMID- 17911456 TI - Harnessing autoimmunity (vitiligo) to treat melanoma: a myth or reality? AB - Melanoma is a highly malignant tumor derived from skin melanocytes (pigment producing cells), which is associated with a significant rate of systemic metastases and death. Various therapeutic approaches for melanoma have been attempted in recent years, including the use of chemotherapy, immunotherapy, and ablative surgical and radiation treatments. However, in many cases these treatments fail as the tumor becomes resistant to the treatment and rapidly spreads and causes death. Reports in the medical literature have documented the unique immunogenic nature of melanoma where antigens, antibodies, and immune complexes seem to play a major role in the course of the disease. Anti-melanoma antibodies can cross-react with antigens on normal melanocytes, therefore causing the appearance of an associated hypopigmentation that resembles vitiligo. Vitiligo is a dermatological disorder characterized by local, dispersed, or diffuse white patches on the skin as a result of the destruction of melanocytes. This disease is believed to be an autoimmune disorder since autoantibodies against membrane components of melanocytes are found in the sera of patients with vitiligo. Melanoma triggers an anti-tumor response in many patients. Unfortunately, such anti-tumor response is insufficient to elicit tumor regression and the tumor continues to proliferate. Since the prognosis of melanoma in patients and animals with vitiligo is more favorable than in the general population, it was hypothesized that sera from patients with vitiligo may react against melanoma cells. Such studies have demonstrated that exposure of tumor cells to the sera resulted in inhibition of proliferation of the melanoma cells in vitro and in regression of melanoma metastases in mice presumably on account of the presence of the high titer of anti-melanoma antibodies in the sera used in these studies. In this review we discuss the known data and hypothetical assumptions related to the use of vitiligo-associated antibodies against melanoma, as well as characterize the immune mechanisms involved in this process. PMID- 17911457 TI - Intravenous immunoglobulin and cytokines: focus on tumor necrosis factor family members BAFF and APRIL. AB - The presence of natural autoantibodies against cytokines has been reported in healthy individuals. Because circulating cytokines may be implicated in the clinical outcome of numerous diseases, the mode of action of intravenous immunoglobulin (IVIg) (pooled from sera over a thousand normal individuals) may involve immunomodulation of the cytokine network. We review the anti-cytokine effects of IVIg as well as the consequences of IVIg infusions on cytokine production. Furthermore, IVIg exerts therapeutic effects in autoimmune diseases and lymphoid malignancies. These two conditions have in common an overproduction of BAFF (for B-cell-activating factor of the TNF family). The presence of antibodies with BAFF and APRIL (a proliferation-inducing ligand) specificity was investigated. We found that IVIg recognizes BAFF and APRIL and that IVIg binding prevents BAFF from exerting its antiapoptotic effect on B cells. These anti-BAFF IgGs might prevent the deleterious effects of BAFF in B-cell-mediated autoimmune diseases. PMID- 17911458 TI - Role of mycophenolate mofetil in the treatment of lupus nephritis. AB - Mycophenolate mofetil (MMF) is an immunosuppressive drug successfully used for the prevention of acute and chronic rejection of renal allografts, as well as in the therapy of glomerular disorders. We treated three groups of patients with lupus nephritis: the first group of patients had a high histologic activity index (AI), 13.4 +/- 2.34; the second group of patients had a high histologic chronicity index (CI), 6.0 +/- 0.7; and the third group consisted of only two patients, one with low AI (3.5) and another with low CI (1.5). The patients were treated for 2 years. MMF was initiated at a dose of 2 g/daily for the first 6 months and the dose was decreased to 1.5 g/daily for the further 18 months. Steroids, 0.4 mg/kg/day, were the concomitant therapy for the first 6 months, with slow tapering for the further 18 months. Patients with high AI presented significant decrease of serum creatinine after 2 years, 286 +/- 112.95 to 131.2 +/- 44.65 micromol/L. Two of the patients, with acute oligoanuria, were withdrawn from dialysis treatment. Significant improvement was also noted, 6.97 +/- 1.81 to 0.9 +/- 0.31 g/day. Patients with high CI had nonsignificant decrease of serum creatinine, 178.5 +/- 47.73 to 129.25 +/- 22.88 micromol/L, and significant improvement of proteinuria, 4.63 +/- 1.57 to 1.14 +/- 0.39 g/day. The patient with low AI showed recovery of renal function (serum creatinine from 196 to 72 micromol/L) and alleviation of proteinuria, 7.93 to 3.4 g/day. The patient with low CI did not respond to the therapy and renal function slowly worsened. MMF has emerged as a promising therapeutic approach for both the induction and maintenance phase in patients with lupus nephritis. PMID- 17911459 TI - Impact of steroids on hepatitis C virus replication in vivo and in vitro. AB - Chronic hepatitis C virus (HCV) infection is the leading indication for liver transplantation. Transplantation outcome is often compromised by a rapid re infection of the graft. Several factors have been implicated in the increased severity of recurrence, including steroid-based immunosuppression. Evidence suggests that steroid boluses used to treat acute rejection are associated with an increase in HCV viral load and the severity of recurrence. Two possible mechanisms for a steroid-mediated effect on HCV viral loads can be postulated, the first being a direct effect of steroids on the virus by enhancing its replication. The second, an indirect effect due to the suppression of the HCV immune response, allows unrestricted HCV replication. To investigate the direct effect of steroids on HCV replication, dexamethasone (Dex) and prednisolone (Pred) were tested in an in vitro replicon model. HCV replication was assessed on the basis of luciferase reporter expression (luminescence) and HCV RNA (RT-PCR). At clinically relevant concentrations (1-10 nM), treatment with both Dex and Pred did not enhance, but resulted in a slight reduction of relative luciferase activity (HCV replication), which was independent of increased cellular protein content and reduced cell proliferation. This minor reduction of HCV replication was confirmed by RT-PCR showing more than 41% reduction in HCV RNA levels. In conclusion, despite clinical evidence that the use of steroids aggravates recurrence of HCV, our in vitro study suggests that there is no direct stimulatory effect of steroids on the replication of HCV. As such, the increased viral loads after high-dose steroid treatment are more likely due to a downregulation of the immune response. In such patients, a dampened immune response allows viruses like HCV to replicate free of immune-mediated killing of their host cells. When a change occurs, such as a tapering or an alteration of immunosuppressant drugs, the immune system reinitiates and vigorously attempts to control the virus, resulting in acceleration of liver damage. Therefore, either steroid avoidance or maintaining low levels, coupled with a slow tapering of corticosteroids, may be beneficial to HCV-infected transplantation recipients. PMID- 17911460 TI - Therapeutic challenges for systemic sclerosis: facts and future targets. AB - Pulmonary arterial hypertension (PAH) is an important cause of death in systemic sclerosis (SSc), despite the improvement of therapies. An early diagnosis and the use of drugs interfering with the main pathogenic pathways of PAH is pivotal for the improvement of prognosis in primary PAH and PAH secondary to autoimmune rheumatic diseases, mainly SSc. Lately, new specific therapies have been developed targeting prostacyclin, endothelin, and nitric oxide pathways, the major pathogenic pathways leading to endothelial dysfunction in PAH. Epoprostenol improved life expectancy of patients with primary and secondary PAH, but its continuous intravenous administration requires experienced centers. More stable analogues of prostacyclin, administrated by intravenous (iloprost, treprostinil), subcutaneous, inhalatory (treprostinil, iloprost), and oral route (Beraprost) have shown efficacy in PAH. Bosentan, the first oral endothelin receptor antagonist (with affinity for endothelin A and B receptors) improves exercise function and survival in PAH, both primary and secondary to autoimmune rheumatic diseases. This is confirmed also for Sitaxsentan and Ambrisentan, selective A receptor antagonists. Because of its short half-life and systemic side effects, short-term NO inhalation is used only in short-term management of PAH in critically ill adults. Inhibitors of NO degradation, such as sildenafil, a phosphodiesterase (PDE) type 5 inhibitor, improved functional and hemodynamic parameters without significant side effects. Vardenafil and taladafil, longer acting PDE inhibitors, also have vascular pulmonary selectivity. All these drugs may be used in combination, to maximize their clinical benefit not only in patients unresponsive to single drugs, but also potentially as initial therapy of PAH. PMID- 17911461 TI - Autologous stem cell transplantation for severe autoimmune diseases: a 10-year experience. AB - The first autologous hematopoietic stem cell transplantation in Europe for a patient with severe refractory systemic lupus erythematosus (SLE) was performed in Genoa in 1996. Since then, 32 patients with a wide spectrum of autoimmune diseases (ADs) received autologous transplants, 22 of them with multiple sclerosis (MS). There were no fatal adverse events. All patients had complete or very good partial remissions, but relapses were frequent, especially in SLE, though never as aggressive as pretransplant. The mechanism of action of this intervention remains not completely understood, as briefly discussed here. PMID- 17911463 TI - Anti-TNF-alpha therapy in seven patients with Behcet's uveitis: advantages and controversial aspects. AB - Behcet's disease (BD) is a chronic, relapsing, multisystem disease. In some patients, ocular involvement can lead to severe vision impairment despite immunosuppressive therapy. Since high levels of circulating TNF-alpha have been found both in peripheral blood and aqueous humor of patients with active BD, we evaluated the efficacy of anti-TNF-alpha therapy in seven patients with severe ocular involvement resistant to previous treatment. Seven patients with sight threatening relapsing uveitis refractory to immunosuppressive regimens received intravenously infliximab, at a dose of 3-5 mg/kg, on week 0-2-4 and then every 6 8 weeks, in combination with low-dose prednisone and methotrexate or azathioprine. Efficacy was assessed in terms of number and severity of relapses of posterior uveitis, visual acuity, and reduction of corticosteroids and immunosuppressive drugs. After a mean follow-up period of 23 months, the total number of relapses dropped to 6, compared to the 21 observed in an equivalent period of time before treatment. The visual acuity improved in 4 eyes, while it remained stable in 9. Therapy with infliximab considerably reduced the required daily dose of both corticosteroids and immunosuppressive drugs. In our experience infliximab proved to be safe and effective in controlling both the number and intensity of cases of posterior uveitis and the extraocular manifestations of BD. It also allowed a reduction of corticosteroids and immunosuppressive drugs required to control the disease. However, ocular and systemic manifestations tended to recur after drug withdrawal or when the interval between infliximab courses was longer than 8 weeks. Moreover, infliximab administration is costly and requires hospital admission. PMID- 17911462 TI - Natalizumab in the treatment of patients with multiple sclerosis: first experience. AB - Multiple sclerosis (MS) usually develops in young adults with a complex predisposing genetic background. Polymorphisms in the gene for chemokine receptor CCR5 have been proposed to confer susceptibility to or protection from MS. Study of molecules participating in the inflammatory process contributed to the development of a new humanized monoclonal antibody, natalizumab, aimed at the adhesive molecule VLA-4. Natalizumab (Biogen Idec/Elan) went through successful clinical studies and its clinical testing was also carried out in the Czech Republic. Twenty-one patients with MS were included in the AFFIRM study (2-year, placebo-controlled study and consecutive 7-month unblinded natalizumab treatment); immunophenotyping of the cerebrospinal fluid (CSF)- CD4+CCR5+CXCR3+ lymphocytes, using flow cytometer FACSCalibur and monoclonal antibodies (BD Biosciences), was done at the end of natalizumab treatment and 1 year after the therapy withdrawal. Compared to MS patients receiving other therapy, the patients treated with natalizumab had statistically significantly (P < 0.0001) higher levels of CCR5+ and lower levels of CD4+ T lymphocytes in CSF, whereas the levels of CXCR3+ lymphocytes were almost the same as in other patients. CCR5-positive CSF lymphocytes decreased 1 year after treatment withdrawal. Natalizumab treatment alters the percentage of CCR5+ and CD4+ cells in CSF. In view of the excellent temporary clinical results of the therapy, which are yet to be assessed in the course of a longer time period, our results show a possible explanation for the therapeutic success of this drug as well as for the development of progressive multifocal leukoencephalopathy. PMID- 17911464 TI - Vascular adhesion protein-1 as a potential therapeutic target in liver disease. AB - Vascular adhesion protein-1 (VAP-1) is a homodimeric, transmembrane sialoglycoprotein, and amine-oxidase enzyme constitutively expressed by hepatic endothelial cells, and as a soluble protein in serum (sVAP-1). VAP-1 mediates leukocyte adhesion and migration in an enzyme activity-dependent manner. We wished to determine whether VAP-1 blockade reduces leukocyte recruitment in inflammatory liver disease, and the mechanism by which this occurs. Our results show that VAP-1 is upregulated in the liver and serum of patients with inflammatory liver disease. Expression is maintained on hepatic sinusoidal endothelial cells (HSECs) isolated from explanted livers. Blockade of VAP-1 activity modestly decreases migration of normal lymphocytes across HSECs but has significant effects on the migration of peripheral blood lymphocytes (PBLs) and liver-derived lymphocytes across HSECs. Engagement of VAP-1 results in PI3-kinase dependent NF-kappaB activation and increased chemokine and adhesion molecule expression. Thus complex mechanisms regulate VAP-1-mediated recruitment of leukocytes. Direct binding to endothelial VAP-1 protein, indirect enzyme dependent activation of other endothelial adhesive pathways, and activation of leukocyte by VAP-1 ligand occupancy all contribute to adhesion. The restricted expression of VAP-1 and increased production of sVAP-1 in inflammatory liver disease confirm the validity of this molecule as a therapeutic target. PMID- 17911465 TI - Intravenous immunoglobulins in autoimmune and inflammatory diseases: a mechanistic perspective. AB - Initially used for the treatment of immunodeficiencies, intravenous immunoglobulins (IVIg) have increasingly been used as immunomodulatory agents in autoimmune and inflammatory disorders. The mode of action of IVIg is enigmatic, probably involving Fc-dependent and/or F(ab')2-dependent nonexclusive mechanisms of action. IVIg broadly interacts with the different components of the immune system: cytokines, complement, Fc receptors, and several cell surface immunocompetent molecules. IVIg has also an impact on effector functions of immune cells. These mechanisms of action of IVIg reflect the importance of natural antibodies in the maintenance of immune homeostasis. We discuss here the recent advances in the understanding of immunoregulatory effects of IVIg, and we pointed out the need for new strategies to overcome the predicted increasing worldwide shortage of IVIg. PMID- 17911466 TI - Intravenous immunoglobulin and autoimmune disease. AB - Intravenous immunoglobulin (IVIg) has been found to have a multitude of uses. However, IVIg is a pooled blood product and as a result a limited resource. At replacement doses (200-400 mg/kg/month) it is of critical utility in the treatment of primary and secondary antibody deficiencies. High-dose immunoglobulin (hdIVIg) given at doses of up to 2 g/kg/day has immunomodulatory action mediated via a number of different effects. First used in the 1980s for the treatment of idiopathic thrombocytopenic purpura, hdIVIg has found uses in a wide range of autoimmune conditions, though for many conditions the evidence base lacks formal randomized controlled trials (RCTs). This article will detail the issues regarding the manufacture and clinical aspects of administration of hdIVIg and its uses, especially with regards to the treatment of autoimmune disease. PMID- 17911467 TI - Mycophenolate mofetil in lupus glomerulonephritis: effectiveness and tolerability. AB - Mycophenolate mofetil (MMF) is an immunosuppressive agent initially used in the treatment of transplant recipients. MMF has been used in renal, heart, and liver transplantation, where it seems more effective than other immunosuppressive regimens in reducing the incidence of acute rejection episodes. MMF has a variety of immunosuppressive effects, including selective suppression of T and B lymphocyte proliferation, and has been more recently used in many autoimmune inflammatory conditions. Systemic lupus erythematosus (SLE) is an autoimmune disease that can potentially involve any organ or system of the human body. Glomerulonephritis (GLN) has been recognized as the most frequent severe manifestation of SLE, leading to poor long-term prognosis. In the treatment of lupus GLN, several therapeutic approaches, all including immunosuppressive drugs, such as cyclophosphamide, azathioprine, or cyclosporine A, have been used. The short- and long-term toxicity of these drugs limits their use in a substantial number of patients. Over the last few years, MMF has emerged as an alternative therapeutic regimen in lupus GLN, mainly for patients refractory to other therapies. These studies have shown that it is highly effective and generally well tolerated. PMID- 17911468 TI - Are neuropeptides important in arthritis? Studies on the importance of bombesin/GRP and substance P in a murine arthritis model. AB - Interference with the effects of neuropeptides may be of potential therapeutic value for the treatment of rheumatoid arthritis (RA). Two neuropeptides that can be discussed in this context are bombesin/gastrin-releasing peptide (BN/GRP) and substance P (SP). In order to obtain new information on the possible importance of these two peptides, the patterns of immunohistochemical expression of BN/GRP and SP and their related receptors in the mouse knee joint from healthy and arthritic mice were examined. Positive staining for GRP receptor and the SP preferred receptor (the neurokinin-1 receptor [NK-1 R]) was observed in articular chondrocytes. On the whole, there was a decrease in immunoreactions for both the GRP- and the NK-1 receptors in the articular chondrocytes in joints exhibiting severe arthritis. Staining for BN/GRP and GRP receptor was seen in the inflammatory infiltrates of the arthritic joints. New evidence for the occurrence of marked effects of BN/GRP concerning both the articular chondrocytes and the inflammatory process is obtained in this study. With these findings and previous observations of neuropeptide expression patterns and functions we discuss the possibility that interventions with the effects of BN/GRP, SP, and other neuropeptides might be worthwhile in RA. PMID- 17911469 TI - A long-term follow-up after cardiac transplantation in a lupus patient: case report and review of the literature. AB - Heart or heart-lung transplantations have only rarely been performed in patients with systemic lupus erythematosus (SLE), who like other patients with multi system autoimmune diseases are traditionally excluded from consideration for such transplantations. In view of the limited experience with heart transplantation in these patients, we report the successful transplantation outcome in a lupus patient and review the literature in relation to graft and recipient conditions. PMID- 17911470 TI - Safety of cyclosporin A in HCV-infected patients: experience with cyclosporin A in patients affected by rheumatological disorders and concomitant HCV infection. AB - Because of the relatively high prevalence of both hepatitis C virus (HCV) infection and autoimmune disorders (ADs), it is not rare to encounter in daily clinical practice patients with ADs also carrying HCV. Corticosteroids and/or immunosuppressant drugs are needed to treat ADs, but they place HCV-infected patients at risk of worsening the infection. So, rheumatologists have often refrained from using corticosteroids or immunosuppressants in AD when HCV-RNA is also present. Cyclosporin A (CsA) is an immunosuppressive agent used to treat a wide range of ADs, but there is a large evidences in the literature, both in vitro and in vivo, suggesting that CsA also exerts an inhibitory effect on HCV replication at standard therapeutic dose. Therefore, this evidence has opened new ways to improve the therapy and the prognosis in patients with HCV-related liver diseases, including those with transplants. Recent reports, although limited in number, also suggest the safety of CsA in the treatment of patients with AD and concomitant HCV infection. In this review we also report our personal experience on the combination treatment with CsA and anti-TNF-alpha agents in rheumatoid arthritis. PMID- 17911471 TI - Suppression of experimental autoimmune myasthenia gravis by intravenous immunoglobulin and isolation of a disease-specific IgG fraction. AB - Intravenous immunoglobulin (IVIG) administration has been beneficially used for the treatment of a variety of autoimmune diseases including myasthenia gravis (MG). We have demonstrated that IVIG administration in experimental autoimmune MG (EAMG) results in suppression of disease that is accompanied by decreased Th1 cell and B cell proliferation. Chromatography of pooled human immunoglobulins (IVIGs) on immobilized IgG, isolated from rats with EAMG, results in a complete depletion of the suppressive activity of the IVIG. Moreover, the eluate from this EAMG-specific antibody column retains the immunosuppressive activity of IVIG. This study supports the notion that the therapeutic effect of IVIGs is mediated by an antigen-specific anti-immunoglobulin (anti-idiotypic) activity that is essential for its suppressive activity. PMID- 17911472 TI - Fetomaternal alloimmunization with antenatal glomerulopathies. AB - Fetomaternal alloimmunization with antenatal glomerulopathies (FMAIG) is a recently described alloimmune disorder, which results from maternal antibodies that cross the placenta, bind to fetal glomerular podocytes, and mediate renal disease. The pathogenic antibodies are directed against neutral endopeptidase (NEP). The infant's mother is NEP-deficient and thus she becomes immunized during pregnancy against NEP expressed by placental cells. Because future pregnancies in NEP-immunized mothers are at high risk for the fetus, detection of anti-NEP antibodies in pregnant mothers and antigen-driven therapies including induction of mucosal tolerance, are urgently needed. This ideally requires identification of the pathogenic epitopes born by the antigen. We have recently characterized two linear B cell epitopes on the NEP that are specifically recognized by the mother's antibodies. The identification of these B cell epitopes is useful for diagnostic tests and may lead to future development of new therapeutic approaches based on peptide-specific immune intervention. PMID- 17911473 TI - Attenuation of colon carcinoma tumor spread by intravenous immunoglobulin. AB - The impact of IVIg on metastatic capacity of CT26 murine colon carcinoma cells was studied using in vitro and in vivo methods. IVIg inhibited CT26 cell proliferation and invasion through an extracellular matrix in a dose- and time dependent manner. Systemic treatment of mice with IVIg significantly inhibited metastatic potential of CT26 colon carcinoma cells observed as tumor nodules and lung weight reduction. Treating CT26 cell-implanted rabbit corneas with IVIg led to shrinking and complete disappearance of tumor mass in 10 days. These results provide the evidence that IVIg may be considered as a supportive therapy for inhibition of colon carcinoma tumor spread. PMID- 17911474 TI - Effective therapy with anti-TNF-alpha in patients with psoriatic arthritis is associated with decreased levels of metalloproteinases and angiogenic cytokines in the sera and skin lesions. AB - Tumor necrosis factor-alpha (TNF-alpha) plays a central role in sustaining the inflammatory process in the skin as well as in the joints of patients with psoriasis and psoriatic arthritis. In fact, biological therapies based on monoclonal antibodies against TNF-alpha have been proven to be effective on both the arthropathy and the cutaneous symptoms of the disease. Among the several effects produced by TNF-alpha on keratinocytes there is the induction of expression of MMP-9, a matrix metalloproteinase (MMP) produced mainly by monocytes and macrophages. In this article we refer to the results of a study on the behavior of MMP-9 in the sera and in the lesional skin in association with effective therapy with infliximab. Measurements of TNF-alpha, MMP-2, vascular endothelial growth factor (VEGF), and E-selectin were also performed in the same samples. Eleven psoriatic patients included in a therapeutic protocol based on the administration of infliximab monotherapy were collected before treatment and after 6 and 12 weeks of therapy. Significant decrease of MMP-9 and MMP-2 levels in the sera was associated with clinical improvement and with the decrease of TNF alpha, VEGF, and E-selectin, angiogenic molecules already known to be implicated in the clinical expression of psoriasis. The clinical amelioration of the cutaneous expression of psoriasis was significantly associated with the decrease of MMP-9, TNF-alpha, and E-selectin levels, spontaneously released by lesional biopsy samples before and after therapy, measured in the culture supernatants by immunoenzymatic assays. In addition, significant correlations were found between the clinical score and TNF-alpha, MMP-9, and E-selectin lesional production. MMP 9 levels were significantly correlated with those of TNF-alpha. Our findings show the existence of a direct relationship between MMP-9 and TNF-alpha production, strongly suggesting that MMP-9 may play a key role in the skin inflammatory process in psoriasis, while a different role may be attributed to MMP-2. PMID- 17911476 TI - Applying embryo-derived immune tolerance to the treatment of immune disorders. AB - Pregnancy is a unique immune state in which both mother and embryo/fetus tolerate and interact with one another through term, without interfering with the mother's native immunity. When the embryo is viable, it initiates maternal immune tolerance (IT)--ut not immune suppression. The balance is complex: some immune disorders are ameliorated during pregnancy, despite the presence of an "inflammatory" environment. We have identified a pregnancy viability biomarker- preimplantation factor (PIF)--secreted only by viable embryos, which helps to initiate this maternal tolerance and uterine receptivity. Using anti-PIF antibodies, we have detected PIF in the fetus and placenta. Beyond enhancing uterine receptivity locally, PIF has also been detected in maternal circulation, and may promote peripheral tolerance. In the fetus, PIF may help avoid a "graft versus-host"-type reaction PIF exhibits unique potent immune-modulatory effects and its synthetic analogue has been shown to exert significant protection in diverse immune scenarios. Nontoxic, low-dose, short-term PIF administration has led to long-term effects in preclinical models of multiple sclerosis (MS), juvenile diabetes mellitus (JDM), and graft-versus-host disease (GVHD), in a manner enabling its translation into a clinical setting. Further investigation of this compound is warranted. PMID- 17911475 TI - Treatment of BXSB-Yaa mice with IL-21R-Fc fusion protein minimally attenuates systemic lupus erythematosus. AB - Interleukin-21 (IL-21) is a pleiotropic cytokine whose function is only now being unraveled. Abundant evidence indicates that activated CD4 T cells are the primary, if not the only, source of IL-21. While it is clear that IL-21 is actively transcribed by naive activated T cells, recent studies have shown that IL-21 potentially promotes a developmental shift of naive T cells toward the Th2 phenotype. BXSB-Yaa mice develop an autoimmune syndrome similar to systemic lupus erythematosus (SLE), affecting males earlier than females on account of the presence of the Yaa (Y-linked autoimmune acceleration) locus. Previous results indicate the elevation of IL-21 expression by BXSB-Yaa mice at an age when the early characteristics of autoimmune processes first become evident. We set out to determine whether IL-21 was necessary for disease progression in BXSB-Yaa mice. Mice were treated for 24 weeks with soluble IL-21R-Fc in order to therapeutically neutralize the IL-21 present. The results overall suggest a biphasic effect of IL 21, negatively influencing survival early on and positively influencing survival at later stages. We propose that IL-21 exerts a pleiotropic effect in which it promotes the protective effects of CD8+ suppressor cells in the early disease phase and then promotes the humoral components of SLE in the later disease stages. This experiment provides preliminary evidence for a role of IL-21 in modulating the severity of SLE in BXSB-Yaa mice. PMID- 17911477 TI - A modified vaccination technique for the prevention and treatment of an experimental autoimmune kidney disease. AB - The main purpose of this article is to introduce a promising new vaccination technique and to outline its efficacy and safety as demonstrated in an experimental autoimmune kidney disease. We have found that antigen (AG)-specific downregulation and/or upregulation of immune responses can be achieved by injections of immune complexes (ICs) which contain prepackaged information. This result is attained with the new vaccination method, a method developed in our laboratory which we have called "modified vaccination technique" (MVT). This MVT not only enables the prevention of pathogenic autoimmune events leading to the development of an experimental autoimmune kidney disease; it also allows, with equal effectiveness, therapeutic intervention to terminate the disease. With an injected IC containing predetermined immune response-inducing components, the process effectuates a specific antibody information transfer conferring advantages that go beyond its prophylactic and therapeutic applicability. Its specificity can induce a precise immune response to correct mishaps, for example, in conditions where the immune system overreacts to an autologous antigen or fails to recognize unwanted self (as in autoimmune disorders, cancer, etc.) Preformed ICs are nontoxic and nonirritant, evoke a predetermined antibody response without the use of adjuvants, cause no disturbance in the overall regulatory function of the immune system, and produce no side effects. We firmly believe that proper implementation of the MVT will be able to induce and maintain specific preventive and/or curative responses in a way that is both natural and more effective in patients with chronic ailments presently treatable only with drugs. PMID- 17911478 TI - A new orally bioavailable synthetic androstene inhibits collagen-induced arthritis in the mouse: androstene hormones as regulators of regulatory T cells. AB - Dehydroepiandrosterone (DHEA) has attracted much interest because of its many antiaging, metabolic and immune-modulating effects in rodents. Synthetic derivatives, such as 5-androstene-16alpha-fluoro-17-one (HE2500) and certain natural metabolites also provide benefit in various animal models of autoimmune and metabolic diseases. But, like DHEA, low potency and low oral bioavailability suggested limited usefulness of these compounds in humans. We hypothesized that HE3286, a novel 17-ethynyl derivative would be orally bioavailable, more potent, and chemically more useful in man than its parent compound. We found that on a dose/mass basis, HE3286 demonstrated up to 25% oral bioavailability in mice. In the DBA mouse model of collagen-induced arthritis (CIA), animals receiving oral treatment with HE3286 (50 mg/kg), beginning at onset of disease, significantly decreased CIA peak scores and daily severity of arthritis scores. Benefit was associated with decreases in: (1) production of TNF-alpha, IL-6, and IL-17; and (2) decreases in joint inflammation, erosion, and synovial proliferation as judged by histological analysis. HE3286 was not found to be immune suppressive in any of the classical models tested, including mitogen-induced proliferation, delayed-type hypersensitivity, or mixed lymphocyte reaction. Instead, benefit was associated with increases in numbers and function of CD4+CD25+FOXp3+CD127- regulatory T cells (T reg). To our knowledge, this is probably the first study to report that an orally bioavailable synthetic analogue of DHEA can ameliorate ongoing disease in a CIA mouse model with relevance to rheumatoid arthritis (RA) and to correlate that finding with decreases in proinflammatory cytokines and increases in T reg cells. Hormones targeting T reg cells hold the intriguing potential to treat autoimmune, infectious, and neoplastic diseases. PMID- 17911479 TI - Mechanism of action of IFN-beta in the treatment of multiple sclerosis: a special reference to CD73 and adenosine. AB - IFN-beta treatment reduces the relapse rate in multiple sclerosis (MS), but the exact mechanism of action of the drug has remained elusive. CD73 (ecto-5' nucleotidase) is an ectoenzyme, which produces adenosine from adenosine monophosphate (AMP) precursor by enzymatic dephosphorylation. AMP is known to be abundantly present at sites of inflammation, and more importantly adenosine, the product of CD73, is known to possess both anti-inflammatory and neuroprotective activity. Our preliminary work has shown that IFN-beta increases the expression of ecto-5'-nucleotidase on endothelial cells (ECs) both in vitro and after systemic treatment of MS patients in vivo. In the majority of MS patients also an increase in the soluble serum CD73 was noted after IFN-beta treatment. Importantly, this correlated with the clinical outcome. CD73 expression on central nervous system (CNS) microvasculature was confirmed with stainings of frozen tissue sections of MS brain samples taken at autopsy. Adenosine, a known neuroprotective agent, might contribute to the beneficial effects of IFN-beta on MS. PMID- 17911480 TI - Impact of treatment with infliximab on serum cytokine profile of patients with rheumatoid and psoriatic arthritis. AB - This article analyzes the serum cytokine profile of a nonrandomized group of patients with rheumatoid arthritis (RA) and psoriatic arthritis (PsA) who are destined to be treated with infliximab following failure after failure of different disease-modifiying antirheumatic drugs (DMARDs). Serial serum samples were collected from 11 patients with refractory RA, three with PsA and one with undifferentiated spondyloarthropathy. All were treated with the antitumor necrosis factor (TNF)alpha agent, infliximab, after failing to sustain a clinical remission with conventional DMARDs. Blood samples were obtained at different phases of their therapy. Serum levels of tumor necrosis factor (TNF)alpha, interferon (IFN)gamma, interleukin (IL)-1beta, IL-6, sIL-2R, IL-10, and IL-1 receptor antagonist (IL-1RA) were determined by commercial ELISA kits. Interestingly, only eight of the 11 patients with RA had elevated TNFalpha serum levels (at least once in their serial measurements). Only one was unresponsive to therapy and despite anti-TNFalpha therapy her serum TNFalpha levels remained extremely high. Two RA patients who responded to infliximab had normal TNFalpha serum levels prior to and following infliximab administration. One RA patient improved after infliximab therapy despite unrelenting high serum levels of TNFalpha, IL-6, and sIL-2R. Patients with active PsA who responded to infliximab therapy had sustained high serum TNFalpha levels. In an unselected population of RA and PsA patients, we noticed diverse patterns of serum cytokine profiles. These results imply that the cytokine profiles of RA and PsA are diverse and their pathogenesis is heterogeneous. PMID- 17911481 TI - Probiotic supplementation with Lactobacillus casei (Actimel) induces a Th1 response in an animal model of antiphospholipid syndrome. AB - Probiotic fermented milk products have the capacity to modulate many immunological mechanisms. Several attempts have been made to alter the progression of various atopic and inflammatory disorders in which the immune system plays a major role. We studied this issue in an animal model of the antiphospholipid syndrome (APS) by supplementing the animals' daily intake with a probiotic mixture. We studied the effects of nutritional supplementation of a commercial product that consists of 10(8)/ml Lactobacillus casei (Actimel) on Balb/c mice that were immunized with beta-2- glycoprotein (beta2GPI) in order to induce a familiar murine model of APS. As controls, we used similar animals that were fed with either yogurt or sham solution as a supplement. We analyzed the effect of Actimel on the concentrations of interleukin (IL)-10 interferon gamma (IFNgamma) as well as the extent of the primary T cell response to beta2GPI, and the levels of autoantibodies to beta2GPI determined by ELISA. Two weeks after priming (in the hind footpad) of Balb/c mice with beta2GPI, we analyzed the cytokine profile of the animals by measuring the concentration of IL-10 and IFNgamma in the supernatants of lymphocytes that were extracted from the popliteal lymph nodes. Following stimulation with 10 microg/mL of beta2GPI, we noticed significant (P < 0.05) suppression of IL-10 production by the stimulated lymphocytes in the animals fed with Actimel and yogurt in comparison to sham solution (73.42 +/- 29.4, 84.7 +/- 8, 196 +/- 41.62 pg/mL, respectively). Both dairy products enhanced the secretion of IFNgamma from 657 +/- 47.09 pg/mL to 896 +/- 78.1, and 933 +/- 76.7 (P < 0.01), respectively; similarly they also accelerated by a mild degree the level of the T cell primary response to beta2GPI measured by [3H]thymidine incorporation. The level of autoantibodies to beta2GPI was suppressed in mice fed with actimel and yogurt in a significant manner (P < 0.05). Actimel as well as yogurt confer an immunological impact on Balb/c mice immunized with beta2GPI. Actimel was able not only to enhance IFNgamma secretion but also to inhibit IL-10 production. PMID- 17911482 TI - Statins as immunomodulators in systemic sclerosis. AB - Systemic sclerosis (SSc) has the highest case-specific mortality among the rheumatic diseases. Vascular dysfunction and structural wall abnormalities are among the earliest and fundamental alterations in SSc. Statins have a number of immunomodulating effects on vascular wall cells, which may modify the progression of vascular injury. The aim of this study was to evaluate the potential efficacy of statin therapy in ameliorating endothelial dysfunction (ED) in SSc by investigating the effect of statins on some markers that reflect endothelial activation in SSc. Forty patients with SSc were randomized into two groups to receive 6 months' treatment with atorvastatin (n = 20; dose, 40 mg/day) or placebo (n = 20) as an adjuvant to existing therapy. Markers of ED including ET 1, plasma nitrate levels, and thrombomodulin (TM) were evaluated by the enzyme linked immunosorbent assay (ELISA) technique. Fibrinogen, high-sensitivity C reactive protein (hsCRP), ESR, lipid peroxide (LP), and malonylaldehyde (MDA) levels were also assessed. Brachial flow-mediated vasodilatation was assessed by ultrasonography. Patients were studied at base line and after 6 months of statin therapy. After 6 months of therapy, ET-1, ICAM-1, sE-selectin, vWF, fibrinogen, ESR, hsCRP as well as LP and MDA levels declined and NO increased significantly in the statin-treated SSc group when compared to the placebo-treated group. Endothelium-dependent vasodilatation (EDV) improved significantly in the atorvastatin-treated group. The findings of this study demonstrated statin mediated improvements in the endothelial function of SSc patients as well as immunomodulating effects. Statins may thus prove to be an invaluable addition to the therapy of the vasculopathy of SSc. PMID- 17911483 TI - Self-reactivity in the dimeric intravenous immunoglobulin fraction. AB - Therapeutic intravenous immunoglobulin (IVIg) preparations contain antibodies reflecting the cumulative antigen experience of the donor population. IVIg contains variable amounts of monomeric and dimeric IgG, but there is little information available on their comparative antibody specificities. We have isolated highly purified fractions of monomeric and dimeric IgG by size-exclusion chromatography. Following treatment of all fractions at pH4, analyses by immunodot and immunocytology on human cell lines showed a preferential recognition of autoantigens in the dimeric IgG fraction. Investigation of the HEp 2 cytoplasmic proteome by 2D-PAGE, Western blot, and subsequent identification of IVIg reactive spots by mass spectrometry (LC-MS/MS) showed that IVIg recognized only a restricted set of the total proteins. Similar experiments showed that more antigens were recognized by the dimeric IgG fraction, especially when the dissociated dimer fraction was used, as compared to its monomeric counterpart. These observations are consistent with idiotype-anti-idiotype masking of auto specific Abs in the dimeric fraction of IVIg. PMID- 17911484 TI - Effects of intravenous immunoglobulins on peripheral blood mononuclear cell activation in vitro. AB - The therapeutic effects of intravenous immunoglobulins (IVIGs) on different chronic inflammatory and autoimmune diseases is well appreciated, though clinical studies with high-evidence levels are largely missing. Similar to the broad spectrum of diseases and their underlying etiopathogenic background, the mechanisms of action seem heterogenous and multifold. Several studies addressing in vitro and in vivo effects of IVIG on various immunological parameters have been described with partly contradictory results. In this study immunoglobulins and stabilizers present in commercial IVIG preparations were studied in regard to the in vitro proliferation and cytokine production of peripheral blood mononuclear cells when stimulated with phytohemagglutinin (PHA), interleukin 2, and tetanus toxoid. Whereas the immunoglobulins stimulate the proliferation of PBMCs and decrease IFNgamma secretion, stabilizers of IVIG seem to inhibit the proliferation of PBMCs while increasing the secretion of IFN gamma. These effects have to be taken into account when balancing the impact of IVIG dosage and infusion intervals and relating them to clinical side effects and therapeutic efficacy. PMID- 17911486 TI - The cover. Drug Jar for Theriac. PMID- 17911487 TI - A piece of my mind. Knowledge is power. PMID- 17911488 TI - Expert panels weigh bisphenol-A risks. PMID- 17911489 TI - New asthma guidelines released. PMID- 17911490 TI - Dual antithrombotic therapy's increased risks not always offset by benefit. PMID- 17911491 TI - Preoperative hematocrit levels and outcomes after noncardiac surgery. PMID- 17911492 TI - Preoperative hematocrit levels and outcomes after noncardiac surgery. PMID- 17911493 TI - Clinical trial registries. PMID- 17911494 TI - Clinical trial registries. PMID- 17911495 TI - Dementia and activities of daily living. PMID- 17911496 TI - Sex differences in the use of implantable cardioverter-defibrillators for primary and secondary prevention of sudden cardiac death. AB - CONTEXT: Previous studies of sex differences in the use of implantable cardioverter-defibrillators (ICDs) predate recent expansions in Medicare coverage and did not provide patient follow-up over multiple years. OBJECTIVE: To examine sex differences in ICD use for primary and secondary prevention of sudden cardiac death. DESIGN, SETTING, AND PARTICIPANTS: Analysis of a 5% national sample of research-identifiable files obtained from the US Centers for Medicare & Medicaid Services for the period 1991 through 2005. Patients were those aged 65 years or older with Medicare fee-for-service coverage and diagnosed with acute myocardial infarction and either heart failure or cardiomyopathy but no prior cardiac arrest or ventricular tachycardia (ie, the primary prevention cohort [n = 65,917 men and 70,504 women]), or with cardiac arrest or ventricular tachycardia (ie, the secondary prevention cohort [n = 52,252 men and 47,411 women]), from 1999 through 2005. MAIN OUTCOME MEASURES: Receipt of ICD therapy and all-cause mortality at 1 year. RESULTS: In the 2005 primary prevention cohort, 32.3 per 1000 men and 8.6 per 1000 women received ICD therapy within 1 year of cohort entry. In multivariate analyses, men were more likely than women to receive ICD therapy (hazard ratio [HR], 3.15; 95% confidence interval [CI], 2.86-3.47). Among men and women alive at 180 days after cohort entry, the hazard of mortality in the subsequent year was not significantly lower among those who received ICD therapy (HR, 1.01; 95% CI, 0.82-1.23). In the 2005 secondary prevention cohort, 102.2 per 1000 men and 38.4 per 1000 women received ICD therapy. Controlling for demographic variables and comorbid conditions, men were more likely than women to receive ICD therapy (HR, 2.44; 95% CI, 2.30-2.59). Among men and women alive at 30 days after cohort entry, the hazard of mortality in the subsequent year was significantly lower among those who received ICD therapy (HR, 0.65; 95% CI, 0.60 0.71). CONCLUSION: In the Medicare population, women are significantly less likely than men to receive ICD therapy for primary or secondary prevention of sudden cardiac death. PMID- 17911497 TI - Sex and racial differences in the use of implantable cardioverter-defibrillators among patients hospitalized with heart failure. AB - CONTEXT: Practice guidelines recommend implantable cardioverter-defibrillator (ICD) therapy for patients with heart failure and left ventricular ejection fraction of 30% or less. The influence of sex and race on ICD use among eligible patients is unknown. OBJECTIVE: To examine sex and racial differences in the use of ICD therapy. DESIGN, SETTING, AND PATIENTS: Observational analysis of 13,034 patients admitted with heart failure and left ventricular ejection fraction of 30% or less and discharged alive from hospitals in the American Heart Association's Get With the Guidelines-Heart Failure quality-improvement program. Patients were treated between January 2005 and June 2007 at 217 participating hospitals. MAIN OUTCOME MEASURES: Use of ICD therapy or planned ICD therapy at discharge. RESULTS: Among patients eligible for ICD therapy, 4615 (35.4%) had ICD therapy at discharge (1614 with new ICDs, 527 with planned ICDs, and 2474 with prior ICDs). ICDs were used in 375 of 1329 eligible black women (28.2%), 754 of 2531 white women (29.8%), 660 of 1977 black men (33.4%), and 2356 of 5403 white men (43.6%) (P < .001). After adjustment for patient characteristics and hospital factors, the adjusted odds of ICD use were 0.73 (95% confidence interval, 0.60 0.88) for black men, 0.62 (95% confidence interval, 0.56-0.68) for white women, and 0.56 (95% confidence interval, 0.44-0.71) for black women, compared with white men. The differences were not attributable to the proportions of women and black patients at participating hospitals or to differences in the reporting of left ventricular ejection fraction. CONCLUSIONS: Less than 40% of potentially eligible patients hospitalized for heart failure received ICD therapy, and rates of use were lower among eligible women and black patients than among white men. PMID- 17911498 TI - PSA failure following definitive treatment of prostate cancer having biopsy Gleason score 7 with tertiary grade 5. AB - CONTEXT: In 2005, the International Society of Urologic Pathology consensus conference recommended that men with biopsy Gleason score 3 + 4 or 4 + 3 prostate cancer and tertiary pattern 5 should have their cancer classified as Gleason score 8 or 9, respectively. Yet, the management of men with Gleason score 7 vs 8 or 9 prostate cancer differs. OBJECTIVE: To compare the prognostic significance of Gleason score 7 with tertiary grade 5 vs other Gleason scores with respect to time to prostate-specific antigen (PSA) failure in men with prostate cancer. DESIGN, SETTING, AND PATIENTS: From 1989 to 2005, 2370 men with clinical tumor category 1c to 3b, node-negative, and nonmetastatic prostate cancer underwent definitive therapy with surgery or radiation therapy with or without hormonal therapy. A pathologist with expertise in genitourinary cancers assigned Gleason scores to the prostate needle biopsy specimens. Cox regression was used to assess whether a significant association existed between the presence of tertiary grade 5 in men with Gleason score 7 disease and time to recurrence compared with men with Gleason score 7 without tertiary grade 5, Gleason score 5 to 6, or 8 to 10 disease, adjusting for known prognostic factors and treatment. MAIN OUTCOME MEASURE: Time to PSA failure. RESULTS: Men with Gleason score 7 and tertiary grade 5 disease had a significantly shorter time to PSA failure than men with 7 without tertiary grade 5 (median time, 5.0 vs 6.7 years, respectively; adjusted hazard ratio (HR), 0.56; 95% confidence interval [CI], 0.32-0.97; P = .04) or score of 6 or less (median time, 15.4 years; adjusted HR, 0.24; 95% CI, 0.13 0.43; P < .001). However, a significant difference was not observed when these men were compared with men with Gleason score 8 to 10 disease (median time, 5.1 years; adjusted HR, 0.96; 95% CI, 0.54-1.71; P = .90). CONCLUSION: In this study population, men with prostate cancer having biopsy Gleason score 7 and tertiary grade 5 had a higher risk of PSA-failure when compared with men with Gleason score 7 without tertiary grade 5 and had a comparable risk with men with Gleason score 8 to 10. PMID- 17911499 TI - Effect of perindopril on large artery stiffness and aortic root diameter in patients with Marfan syndrome: a randomized controlled trial. AB - CONTEXT: Aortic stiffness is increased in Marfan syndrome contributing to aortic dilatation and rupture, the major cause of premature death in this population. Angiotensin-converting enzyme inhibitors have been shown to reduce arterial stiffness. OBJECTIVE: To determine whether perindopril therapy reduces aortic stiffness and attenuates aortic dilatation in patients with Marfan syndrome. DESIGN, SETTING, AND PARTICIPANTS: A randomized, double-blind, placebo-controlled trial of 17 patients with Marfan syndrome (mean [SD], 33 [6] years) taking standard beta-blocker therapy, initiated in January 2004 and completed in September 2006, at Alfred Hospital Marfan Syndrome Clinic, Melbourne, Australia. INTERVENTION: Patients were administered 8 mg/d of perindopril (n = 10) or placebo (n = 7) for 24 weeks. MAIN OUTCOME MEASURES: Indices of arterial stiffness were assessed via systemic arterial compliance, and central and peripheral pulse wave velocities. Aortic root diameters were assessed at 4 sites via transthoracic echocardiography. RESULTS: Perindopril reduced arterial stiffness as indicated by increased systemic arterial compliance (mean [SEM], 0.33 [0.01] mL/mm Hg at baseline to 0.54 [0.04] mL/mm Hg at 24 weeks in perindopril group vs 0.30 [0.01] mL/mm Hg to 0.29 [0.01] mL/mm Hg in placebo group, P = .004), and reduced central (7.6 [0.4] m/s to 5.9 [0.3] m/s in perindopril group, P < .001 vs placebo) and peripheral (10.9 [0.4] m/s to 8.7 [0.4] m/s in perindopril group, P < .001 vs placebo) pulse wave velocities. In addition, perindopril significantly reduced aortic root diameters relative to placebo in both end-systole and end-diastole (P<.01 to P < .001 for all comparisons between groups). Although perindopril marginally reduced mean arterial pressure (from 81 [2] mm Hg to 80 [1] mm Hg in perindopril group vs 83 [2] mm Hg to 84 [3] mm Hg in placebo group, P = .004), the observed changes in both stiffness and left ventricular outflow tract diameter remained significant when mean arterial pressure was included as a covariate. Transforming growth factor beta (TGF-beta), which contributes to aortic degeneration in Marfan syndrome, was reduced by perindopril compared with placebo in both latent (59 [6] ng/mL to 45 [3] ng/mL in perindopril group, P = .01 vs placebo) and active (46 [2] ng/mL to 42 [1] ng/mL in perindopril group, P = .02 vs placebo) forms. CONCLUSIONS: Perindopril reduced both aortic stiffness and aortic root diameter in patients with Marfan syndrome taking standard beta-blocker therapy, possibly through attenuation of TGF-beta signaling. Large clinical trials are needed to assess the clinical benefit of angiotensin II blockade in Marfan syndrome. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00485368. PMID- 17911500 TI - A 30-year-old woman with chronic hypertension trying to conceive. AB - Mrs F is a 30-year-old woman with a history of chronic hypertension and possible preeclampsia during her first pregnancy. She is currently trying to conceive and wants to know how her hypertension will affect a future pregnancy and how it should best be managed, both now and during a pregnancy. The management of chronic hypertension before, during, and after a pregnancy is discussed with an emphasis on the goals of treatment and safety of medications during pregnancy and with breastfeeding. Preeclampsia is the most common complication of chronic hypertension in pregnancy and is a particular worry for Mrs F because she may have had it with her prior pregnancy. The current understanding of the pathogenesis of this enigmatic illness is therefore also reviewed, along with its implications for long-term maternal health. PMID- 17911501 TI - Update: a 69-year-old woman with left main coronary artery disease. PMID- 17911502 TI - Abortion politics: clinical freedom, trust in the judiciary, and the autonomy of women. PMID- 17911503 TI - Disparities in use of implantable cardioverter-defibrillators: moving beyond process measures to outcomes data. PMID- 17911504 TI - Caring for women with hypertension in pregnancy. PMID- 17911505 TI - "Gotta' sing! Gotta' diagnose!": a postmortem examination of Rodgers and Hammerstein's medical musical Allegro. PMID- 17911506 TI - JAMA patient page. Grading of prostate cancer. PMID- 17911508 TI - Practice guideline for the performance of physiologic evaluation of extremity arteries. PMID- 17911509 TI - Payer costs in patients undergoing uterine artery embolization, hysterectomy, or myomectomy for treatment of uterine fibroids. AB - PURPOSE: To compare health resource use and medical costs in patients with uterine leiomyomas treated with hysterectomy, myomectomy, or uterine artery embolization (UAE). MATERIALS AND METHODS: Patients who underwent hysterectomy, myomectomy, or UAE for leiomyomas were identified from a nationally representative private payer claims database based on their diagnosis and procedure codes. The study included patients with no prior hysterectomy, myomectomy, or UAE and no previous diagnosis of gynecologic cancer. Health resource use and medical costs were evaluated over a period of 12 months. RESULTS: The study included 2,836 hysterectomy, 704 myomectomy, and 125 UAE patients. Average patient ages were 46 years for hysterectomy, 38 years for myomectomy, and 45 years for UAE (P < .001). Median UAE procedure costs were $5,968, compared with $7,299 for myomectomy (P = .031) and $7,707 for hysterectomy (P < .001). Median total 12-month payer costs were not significantly different among the three procedures ($10,519 for UAE vs $9,652 for myomectomy [P = .372] and $10,044 for hysterectomy [P = .813]). There were no differences in overall hospital admissions or emergency room visits after the procedures. Patients who underwent UAE had greater fibroid-related hospital and physician office use beyond 30 days after treatment (P < .001). During this period, 65.6% of patients treated with UAE had at least one imaging study, versus 37.1% of those treated with myomectomy (P < .001) and 14.1% of those treated with hysterectomy (P < .001). CONCLUSIONS: Procedure costs were significantly lower for UAE versus myomectomy and hysterectomy, but there was no difference in total 12-month payer costs. Postprocedural imaging appears to be a factor in total UAE costs. Further research is needed to better understand the role of imaging studies after UAE. PMID- 17911510 TI - Stent placement for the treatment of portal vein stenosis or occlusion in pediatric liver transplant recipients. AB - PURPOSE: To evaluate the efficacy of stent placement for the treatment of portal vein (PV) stenosis or occlusion in pediatric liver transplant recipients. MATERIALS AND METHODS: Written informed consent was obtained from a legal guardian, and our institutional review board approved this study. Percutaneous (n = 10) or intraoperative (n = 2) stent placement was attempted in 12 pediatric recipients (age range, 6-102 months) via the percutaneous transhepatic or inferior mesenteric vein route. Stents 6-10 mm in diameter were placed. Technical and clinical success, complications, and patency of the PV were retrospectively analyzed. RESULTS: Technical success was achieved in 10 of 12 patients (83%) and clinical success was achieved in eight patients (67%). Eight of the 10 patients in whom technical success was achieved (80%) remained healthy with a patent PV during the 10-58-month clinical follow-up period. One patient with technical success died of acute rejection without recurrent PV complications and another died of acute rejection after stent replacement as a result of an hourglass deformity of a deployed stent with partial thrombosis. No major procedural complications occurred. CONCLUSIONS: Based on this study in a relatively small number of patients, PV stent placement seems to be a safe and effective method for the treatment of posttransplantation PV stenosis or occlusion in pediatric patients. PMID- 17911511 TI - Treatment of high-flow priapism with superselective transcatheter embolization in 27 patients: a multicenter study. AB - PURPOSE: To evaluate the effectiveness and safety of treatment of high-flow priapism (HFP) with superselective transcatheter embolization at nine university hospitals. MATERIALS AND METHODS: Between May 1994 and October 2006, 27 patients underwent superselective embolization of the cavernous artery for HFP. Trauma was apparent in 22 patients, there was self-administered intracavernosal injection for erectile dysfunction in two, and the remaining three did not recall any penile or perineal trauma. The embolic agents used were autologous blood clot (n = 12), gelatin sponge (n = 12), microcoils combined with gelatin sponge (n = 1), polyvinyl alcohol (n = 1), and N-butyl cyanoacrylate (n = 1). Recurrence of priapism and change in erectile function were evaluated during a mean follow-up of 13 months. Differences in results between patients treated with autologous blood clot versus gelatin sponge were statistically analyzed with use of the chi(2) test. RESULTS: In 24 of 27 patients (89%), a single embolization was sufficient for complete resolution of priapism. Repeat embolization was required in two patients (7%), and in the remaining patient (4%), shunt surgery was performed after embolization as a result of HFP coexisting with corporeal venoocclusive dysfunction. Eighteen of 23 patients (78%) who had premorbid normal erectile function showed maintained potency during the follow-up period. There was no significant difference affecting required repeat embolization (P = .537) and change in quality of erection (P = .615) during the follow-up period between the autologous blood clot and gelatin sponge treatment groups. CONCLUSIONS: Superselective transcatheter embolization in the treatment of HFP is effective and ensures a high level of preservation of premorbid erectile function. PMID- 17911512 TI - Impact of tip culture on the management of infected tunneled hemodialysis catheters. AB - PURPOSE: It is common practice to perform culture of the tip of a catheter when a previously indwelling vascular catheter is removed for presumed infection. This study was conducted to determine the effect of hemodialysis catheter tip cultures (CTCs) on patient management. MATERIALS AND METHODS: Patients whose tunneled hemodialysis catheter (TDC) was removed for infection and who had a CTC between July 2001 and June 2006 were identified. The study sample included 242 patients with 330 catheters (109 men, 133 women). Patients' mean age was 58 years (range, 22-94 y). Catheter tip and blood culture (BC) results obtained during the event leading to catheter removal were recorded. Antimicrobial therapy before and after cultures was recorded. Medical records were reviewed for any indication that the treatment plan was influenced by tip culture results. RESULTS: Of 330 catheters, 134 had positive CTC findings; of these, complete records were available for 52 catheters. The treatment plan for patients with suspected catheter-related bloodstream infection (CRBSI) leading to TDC removal did not change in response to semiquantitative CTC results (51 of 52 patients; 98%). In a single case in which CTC results impacted management, this was because of a delay in processing BCs. Negative CTC findings (n = 108) never changed management (ie, resulted in cessation of antibiotic therapy). Seventy-three percent of patients (n = 38) received empiric antibiotics before any culture results. In these cases, the BC susceptibility profiles confirmed the appropriateness of the initial treatment (n = 10) or led to a change (n = 28). CONCLUSIONS: CTC did not alter management in the treatment of TDC infection. Because CTCs increase costs, their routine use should be abandoned in this population. PMID- 17911513 TI - Time to hemostasis after traction removal of tunneled cuffed central venous catheters. AB - PURPOSE: Many patients undergo placement of tunneled cuffed central venous catheters (TCCVCs) for indications including administration of medical therapy and hemodialysis. They are removed when no longer needed or if there is a device complication. There is no consensus regarding the necessity of routine preremoval coagulation studies or platelet count, so this study was performed to determine if abnormal coagulation status affects the time to hemostasis (TH) after traction removal of TCCVCs. MATERIALS AND METHODS: Adult patients referred to our group for removal of a TCCVC placed via a jugular or subclavian route were considered candidates for inclusion. Blood was submitted for evaluation of prothrombin time (PT) and International Normalized Ratio (INR), activated partial thromboplastin time (aPTT), and platelet count. Catheters were removed with the traction technique, and presence of hemostasis was assessed at 5-minute intervals of manual compression. RESULTS: Between November 19, 2001, and April 20, 2004, 179 subjects were enrolled and completed the study. There were 165 subjects in whom TH was within the first 5-minute interval and 14 in whom more than 5 minutes was required. Statistically significant factors associated with prolonged TH were primary diagnosis of end-stage renal disease (P = .005), use of antiplatelet agents (P = .03), and procedure performed by a "low-volume" operator (P = .002). CONCLUSIONS: Routine preremoval evaluation of coagulation parameters is not necessary. Patients who are likely to have abnormal platelet function but not abnormal platelet number appear to be at risk for prolonged TH, but even in those cases, the THs are rarely more than 15 minutes. PMID- 17911514 TI - CT fluoroscopy guided biopsy of small pulmonary and upper abdominal lesions: efficacy with a modified breathing technique. AB - PURPOSE: To characterize a new protocol of computed tomographic (CT) fluoroscopy guided biopsy of the lung and upper abdomen to minimize the intervention time, complication rate, and exposure to ionizing radiation for both the patient and the radiologist. MATERIALS AND METHODS: Fifty patients (23 women, 27 men; mean age, 64.3 years; age range, 36-83 years) with lung (n = 41) or upper abdomen (n = 9) nodules 15 mm or smaller underwent CT fluoroscopy-guided biopsy from November 2005 to October 2006. The mean nodule diameter was 12.6 mm (range, 8-15 mm), the mean depth to skin was 57.3 mm (range, 20-114 mm), and the mean depth of nodules from pleura and/or peritoneum was 18.9 mm (range, 1-77 mm). Histopathologic evaluation of samples was performed on the day of the procedure. A CT fluoroscopy guided biopsy protocol was established as follows: (a) native CT with breath holding at an intermediate respiration level, (b) selection of section position with target nodule and insertion of an 18-gauge coaxial biopsy needle extrapleurally and/or extraperitoneally virtually targeting at nodule, (c) start of CT fluoroscopy (130 kVp, 30 mAs, 5-mm-thick sections) at inspiration level with the patient expiring, (d) stop of CT fluoroscopy when the target nodule reaches the section position, short breath-hold, needle advancement to the target nodule, (e) control of needle position with CT fluoroscopy, and (f) biopsy. RESULTS: The mean total table time was 23.8 minutes (range, 15-41 minutes), the mean duration of CT fluoroscopy was 8.2 seconds (range, 4-23 seconds), and the mean duration of breath-holding--including needle insertion to target nodule and control CT fluoroscopy--was 10.3 seconds (range, 5-15 seconds). There were three minor pneumothoraces that required no further intervention, seven minor pulmonary hemorrhages, three moderate pulmonary hemorrhages with hemoptysis, and one moderate liver hematoma. There were no major complications. The diagnostic accuracy of biopsy samples was 96%. CONCLUSIONS: The presented modification of CT fluoroscopy-guided biopsy of mobile pulmonary and upper abdominal lesions is a rapid and safe procedure, requiring only short exposure to ionizing radiation. PMID- 17911515 TI - A comparison of chemoembolization endpoints using angiographic versus transcatheter intraarterial perfusion/MR imaging monitoring. AB - PURPOSE: Transcatheter arterial chemoembolization (TACE) is an established treatment for unresectable liver cancer. This study was conducted to test the hypothesis that angiographic endpoints during TACE are measurable and reproducible by comparing subjective angiographic versus objective magnetic resonance (MR) endpoints of TACE. MATERIALS AND METHODS: The study included 12 consecutive patients who presented for TACE for surgically unresectable HCC or progressive hepatic metastases despite chemotherapy. All procedures were performed with a dedicated imaging system. Angiographic series before and after TACE were reviewed independently by three board-certified interventional radiologists. A subjective angiographic chemoembolization endpoint (SACE) classification scheme, modified from an established angiographic grading system in the cardiology literature, was designed to assist in reproducibly classifying angiographic endpoints. Reproducibility in SACE classification level was compared among operators, and MR imaging perfusion reduction was compared with SACE levels for each observer. RESULTS: Twelve patients successfully underwent 15 separate TACE sessions. SACE levels ranged from I through IV. There was moderate agreement in SACE classification (kappa = 0.46 +/- 0.12). There was no correlation between SACE level and MR perfusion reduction (r = 0.16 for one operator and 0.02 for the other two). CONCLUSIONS: Angiographic endpoints during TACE vary widely, have moderate reproducibility among operators, and do not correlate with functional MR imaging perfusion endpoints. Future research should aim to determine ideal angiographic and functional MR imaging endpoints for TACE according to outcome measures such as imaging response, pathologic response, and survival. PMID- 17911516 TI - Clinical utility of coaxial reservoir system for hepatic arterial infusion chemotherapy. AB - PURPOSE: To test the feasibility and clinical utility of a reservoir with coaxial catheters (a 2.9-F microcatheter and a 5-F catheter) and a port (ie, coaxial reservoir) that was developed to perform repeated hepatic arterial infusion chemotherapy (HAIC) in patients with unresectable liver neoplasms. MATERIALS AND METHODS: The coaxial reservoir was implanted in 64 patients with unresectable liver neoplasms as a result of difficulty in implanting a conventional reservoir with a 5-F catheter. The 2.9-F microcatheter tip was inserted into the gastroduodenal artery (n = 22), pancreaticoduodenal arcade (n = 20), or peripheral hepatic artery (n = 22) through the 5-F catheter, and a side hole created in the leading end of the microcatheter was oriented toward the proper hepatic artery. Technical success was defined by implantation of the coaxial reservoir and initiation of HAIC. The study endpoint was interruption of HAIC or death. Technical success and early and delayed complications were recorded. RESULTS: The technical success rate was 100%. HAIC was repeated every 1-4 weeks during the mean follow-up period of 14.1 months. Arterial infusion chemotherapy was interrupted in 17 patients (27%) as a result of hepatic arterial occlusion (16%, n = 10), catheter dislocation (3%, n = 2), catheter occlusion (3%, n = 2), wound infection (3%, n = 2), or breakage of the port (2%, n = 1). Patency rates of the hepatic artery were 96%, 82%, and 50% at 6 months, 1 year, and 2 years after reservoir implantation, respectively. CONCLUSION: Implantation of the coaxial reservoir is feasible, safe, and useful in expanding the indication of HAIC to patients with unresectable liver neoplasms. PMID- 17911517 TI - Percutaneous radiofrequency ablation for pulmonary metastases from colorectal cancer: midterm results in 27 patients. AB - PURPOSE: To retrospectively evaluate the midterm outcomes (eg, safety, local efficacy, and survival) after radiofrequency (RF) ablation for pulmonary metastases from colorectal cancer. MATERIALS AND METHODS: Twenty-seven patients (19 men and eight women; mean age, 61.6 years) with 49 pulmonary metastases (mean long axis diameter, 1.5 cm) from colorectal cancer underwent 41 percutaneous computed tomography (CT)-guided RF ablation sessions. Follow-up examinations were performed with CT by using contrast medium administration in all patients; positron emission tomography was performed in five patients. The safety of the procedure, local tumor control, and patient survival were evaluated. Multiple variables were analyzed to determine prognostic factors. RESULTS: Pneumothorax occurred after 20 of the 41 sessions (49%), three of which necessitated chest tube placement. A small pleural effusion was found after six of the 41 sessions (15%). No major hemorrhagic event was observed. None of the patients died due to the procedure. The median follow-up period was 20.1 months (range, 11.2-47.7 months). The primary and secondary technique effectiveness rates were 72% and 85%, respectively, at 1 year, 56% and 62% at 2 years, and 56% and 62% at 3 years. The overall survival rates after RF ablation were 96% at 1 year, 54% at 2 years, and 48% at 3 years. The presence of extrapulmonary metastasis was determined to be a prognostic factor (P = .001). CONCLUSIONS: The midterm outcomes of percutaneous RF ablation for colorectal pulmonary metastases appear promising. The presence of extrapulmonary metastasis had an adverse effect on survival after RF ablation. PMID- 17911518 TI - Matrix metalloproteinase-9 may play a role in recanalization and recurrence after therapeutic embolization of aneurysms or arteries. AB - PURPOSE: Matrix metalloproteinase (MMP)-9 plays various roles in vascular healing and angiogenesis. This study was conducted to determine if MMP-9 is involved in healing or recanalization after therapeutic occlusion of arteries or aneurysms. MATERIALS AND METHODS: Angiographic and pathologic changes were investigated in canine bilateral venous pouch carotid aneurysms embolized with gelatin sponges with or without previous endothelial denudation, a procedure that can prevent recanalization. To assess a potential role of MMP-9, messenger RNA (mRNA) and protein were compared in denuded and nondenuded aneurysms 4, 7, and 14 days after embolization. To assess if MMP-9 is essential to arterial recanalization, transmyocardial angiography and pathologic findings were compared 14 days after carotid occlusion with platinum coils in MMP-9-knockout and wild-type mice. RESULTS: Denudation of the endothelial lining led to improved angiographic results at 3 weeks (P < .001). Neointimal closure of the aneurysm neck was more complete in denuded versus nondenuded aneurysms. Denudation was followed by a decrease in MMP-9 mRNA (86%, P < .05) and protein (30%, P < .05) 7 days after embolization and a decrease in von Willebrand factor compared with nondenuded aneurysms. MMP-9 immunostaining of axial sections from embolized aneurysms confirmed MMP-9-positive endothelialized clefts, which were absent in denuded aneurysms. Transmyocardial angiography and pathologic examination showed recanalization of one of nine coiled carotid arteries of MMP-9-knockout mice, compared with five of seven controls (P = .035). CONCLUSIONS: MMP-9 may play a role in recanalization of arteries after coil occlusion and in recurrences after sponge embolization of aneurysms. PMID- 17911519 TI - Comparison of transcatheter intraarterial perfusion MR imaging and fluorescent microsphere perfusion measurements during transcatheter arterial embolization of rabbit liver tumors. AB - PURPOSE: Transcatheter intraarterial perfusion (TRIP) magnetic resonance (MR) imaging is clinically used in the interventional MR imaging setting to verify distribution of injected embolic or chemoembolic material during liver-directed transcatheter therapies and to monitor reductions in perfusion. The accuracy of this technique remains unknown. In the present study, rabbit VX2 liver tumors were used to test the hypothesis that TRIP MR imaging accurately measures changes in tumor perfusion during transcatheter arterial embolization (TAE), with injection of fluorescent microspheres used as the gold-standard technique. MATERIALS AND METHODS: Five New Zealand White rabbits were used for this study (two donor rabbits and three with VX2 liver tumors). In three rabbits with implanted VX2 liver tumors, catheters were superselectively placed under digital subtraction angiographic guidance into the left hepatic artery supplying the targeted tumor. Fluorescent microspheres were injected into each rabbit's left ventricle before and after TAE. TRIP MR images were obtained at baseline and after embolizations for all rabbits with intraarterial injections of 2.5% gadopentetate dimeglumine solution. Linear regression was used to compare relative reductions in tumor perfusion between TRIP MR imaging and fluorescent microspheres. Results were considered statistically significant at a P value less than .05. RESULTS: There was good correlation between TRIP MR imaging and fluorescent microsphere measurements of reduction in tumor perfusion (r = 0.722, P < .012). CONCLUSIONS: TRIP MR imaging provides accurate semiquantitative measurement of perfusion reduction during TAE in rabbit liver tumors. PMID- 17911520 TI - MR imaging detection of superparamagnetic iron oxide loaded tris-acryl embolization microspheres. AB - PURPOSE: To assess by magnetic resonance (MR) imaging the detectability of superparamagnetic iron oxide (SPIO)-labeled microspheres (MSs) in vitro on gelose, ex vivo in kidneys from embolized sheep, and in vivo in kidneys from embolized pigs. MATERIALS AND METHODS: With various sizes of SPIO-labeled MSs, common neck and pelvic spin-echo and gradient-echo sequences were acquired on a 1.5-T MR unit. SPIO-labeled MSs of four sizes were embedded in a hydrogel as single MSs or in multiple units, or multiplets. Detection rate on MR imaging was assessed according to the real size and number of MSs. SPIO-loaded and unloaded MSs of four sizes were injected into eight sheep kidneys, which underwent MR and pathologic examinations. For each size, the location of MSs in renal vasculature was determined and compared according to the technique used. Kidneys were embolized in pigs with various amounts of MSs in three sizes. MR was performed immediately after embolization and SPIO-labeled MS detection was assessed according to size, organ, and amount injected. Results SPIO-labeled MSs provide a low signal intensity on T1-weighted sequences, without distortion. In vitro, 28% of 100-300-microm single MSs were detected and more than 80% were detected for larger sizes. MS multiplets were all detected in all sizes. Ex vivo, all sizes of MSs were detected by MR imaging in kidneys, whereas control MSs were not observed. Histologic analysis showed that there was no difference in vascular distribution between SPIO-labeled MS and control MSs, and therefore for each caliber (P > .05). Arterial location of SPIO-labeled MSs was the same on MR imaging and histologic analysis. In vivo, SPIO-labeled MS were detected in the kidney vasculature when volumes greater than 1 mL of 100-300-microm or 500-700 microm MSs were injected. Volumes lower than 1 mL SPIO-labeled MSs were hardly detected in kidneys, regardless of MS size. Conclusions SPIO-labeled MSs are detected by MR imaging with common gradient-echo sequences in vitro in gelose and ex vivo and in vivo in kidneys. SPIO-labeled MSs could allow better control of embolization and thereby enhance efficacy and safety of the procedure. PMID- 17911521 TI - Fracture of a nitinol carotid artery stent that caused restenosis. AB - The placement of self-expanding nitinol stents in the carotid artery is rapidly emerging as a minimally invasive alternative in the treatment of cervical carotid artery occlusive disease. Fractures of balloon-expandable stents after placement in the carotid artery have been previously documented. The authors report on a single case of early stent fracture and complete mid-body disruption of a self expanding stent associated with a significant restenosis necessitating repeat intervention after carotid artery stent placement for a de novo atherosclerotic lesion. PMID- 17911522 TI - Double stent technique for the treatment of an internal carotid artery pseudoaneurysm caused by zone III stab injury. AB - A 77-year-old man was transferred to the hospital with swelling of his neck and oropharynx after a stab injury to his oral cavity with pruning shears. Findings at complete neurologic examination were normal. Contrast-enhanced computed tomography (CT) and angiography revealed a pseudoaneurysm at the pharyngeal portion of the right internal carotid artery. Endovascular treatment was undertaken by using the double bare stent technique. The pseudoaneurysm was completely occluded immediately after the procedure. There were no complications. There were no further symptoms or evidence of recurrence of the aneurysm during the 18-month follow-up period. The double bare stent technique is safe and effective for the treatment of zone III carotid artery stab injuries. PMID- 17911523 TI - Effect of C-arm angiographic CT on transcatheter arterial chemoembolization of liver tumors. AB - Rotational C-arm angiographic computed tomography (CT) with a flat-panel radiography unit permits three-dimensional (3D) reconstruction of soft tissues and blood vessels. The usefulness of this C-arm technique during transcatheter arterial chemoembolization (TACE) is unknown. The authors analyzed the role of the C-arm technique in 18 patients with unresectable liver tumors during TACE. The technique altered the catheter position anticipated by attending interventional radiologists in seven of the 18 patients (39%; 95% confidence interval [CI]: 20%, 61%) and improved the diagnostic confidence in the selected catheter position in 14 of the 18 patients (78%; 95% CI: 55%, 91%). The technique provides CT-like images that are useful to interventional radiologists during TACE. PMID- 17911524 TI - Reduction of arteriohepatovenous shunting by temporary balloon occlusion in patients undergoing radioembolization. AB - Radioembolization with yttrium-90 resin microspheres is a treatment option that selectively targets hepatic tumors. One of the primary limiting factors for this therapy is the degree of arteriohepatovenous shunting, as excessive radiation to the lungs may cause radiation pneumonitis. To safeguard patients against this, a technetium Tc 99m macroaggregated albumin scan is performed before treatment to assess the degree of arteriohepatovenous shunting. As lung shunt fraction increases, activity reductions are mandated, with a 20% shunt sufficient to prohibit treatment. Temporary occlusion of shunts may be achieved by placement of balloon catheters in the hepatic veins. This endovascular technique used to reduce arteriohepatovenous shunting allows otherwise untreatable patients to undergo radioembolization. PMID- 17911525 TI - Loop-snare technique for difficult inferior vena cava filter retrievals. AB - Retrievable inferior vena cava (IVC) filters are generally retrieved without difficulties. However, when filters are tilted against the IVC wall, engaging the tip or hook of the filter can be difficult with the use of standard techniques. This report describes an alternative method of successful IVC filter retrieval by creating a wire loop between the filter legs, including the tip of the filter. This loop snare was successfully applied in eight cases after filter retrieval failed with the Recovery Cone or simple snare technique. PMID- 17911526 TI - Use of rigid bronchoscopic forceps in the difficult retrieval of the Gunther Tulip inferior vena cava filter. AB - Retrievable inferior vena cava (IVC) filters may, over time, become incorporated into the wall of the IVC, making subsequent removal difficult. The authors present a case in which a Gunther Tulip filter was placed prophylactically before gastric bypass surgery. The retrieval hook of the filter became incorporated into the wall of the IVC, preventing the filter from being snared. Eventually, the filter was freed from the wall of the IVC and successfully removed by using rigid bronchoscopy forceps; however, the filter was damaged in the process. PMID- 17911527 TI - Persistent urine leak after cryoablation of a renal tumor in a patient with an ileal conduit. AB - Thermal ablation is expanding as a technique to treat small renal masses. Damage to the collecting system is uncommon with either radiofrequency ablation or cryoablation. Early evidence suggests that cryoablation is less damaging to the urinary tract, and investigators have advocated the use of aggressive treatment in central tumors in contact with the renal hilum. The authors report a nonhealing urinary fistula after successful cryoablation of an exophytic upper pole renal mass in a patient with an ileal conduit. The presence of an ileal conduit may present an increased risk of urinary fistula after thermal ablation. PMID- 17911528 TI - Standing or stationary arterial waves of the superior mesenteric artery at MR angiography and subsequent conventional arteriography. PMID- 17911529 TI - Nonocclusive mesenteric ischemia: findings at multidetector CT angiography. PMID- 17911530 TI - Percutaneous vertebroplasty causing an increase in retropulsion of bone fragments. PMID- 17911531 TI - Perforation of a subhepatic retrocecal appendix after radiofrequency ablation of a hepatic metastasis. PMID- 17911532 TI - Peroxide is a key mediator of Bcl-2 down-regulation and apoptosis induction by cisplatin in human lung cancer cells. AB - Susceptibility to apoptosis is an essential prerequisite for successful eradication of tumor cells by chemotherapy. Consequently, resistance to apoptosis has been established as one of the mechanisms responsible for the failure of therapeutic approaches in many types of cancers. In the present study, we investigated the susceptibility of human lung cancer H460 cells to apoptotic cell death induced by cisplatin and determined its regulatory mechanisms. Treatment of the cells with cisplatin induced rapid generation of multiple oxidative species and a concomitant increase in apoptotic cell death. Apoptosis induced by cisplatin was mediated through the mitochondrial death pathway, which requires caspase-9 activation and is regulated by Bcl-2. Cisplatin induced down-regulation of Bcl-2 through a process that involves dephosphorylation and ubiquitination of the protein, which facilitates its degradation by proteasome. This down regulation was inhibited by antioxidant enzymes catalase and glutathione peroxidase (H(2)O(2) scavenger), but not by superoxide dismutase (O(2)(.) scavenger) or deferoxamine (OH. inhibitor). Electron spin resonance and flow cytometric analyses showed the formation of H(2)O(2) along with O(2)(.) and OH. radicals after cisplatin treatment. H(2)O(2) was generated in part by dismutation of O(2)(.) and served as a precursor for OH.. Together, our results indicate an essential role of H(2)O(2) in the regulation of Bcl-2 and apoptotic cell death induced by cisplatin. Because aberrant expression of Bcl-2 has been associated with death resistance of cancer cells to chemotherapy, the results of this study could be used to aid the design of more effective strategies for cancer treatment. PMID- 17911533 TI - Salicylate blocks lipolytic actions of tumor necrosis factor-alpha in primary rat adipocytes. AB - Increased systemic free fatty acids (FFA) impair insulin sensitivity. In obese and diabetic subjects, production of tumor necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine, is elevated. TNF-alpha has a variety of effects by inducing inflammation, decreasing glucose utilization, and stimulating adipocyte lipolysis to release FFA to plasma. High doses of nonsteroidal anti-inflammatory drug salicylates have long been recognized to lower blood FFA and glucose in humans, although the mechanisms are not fully understood. In this report, we show that sodium salicylate at therapeutic concentrations directly blocks TNF-alpha stimulated lipolysis and therefore inhibits FFA release from primary rat adipocytes. To elucidate the cellular basis of this action, we show that salicylate suppresses TNF-alpha-induced extracellular signal-related kinase activation and intracellular cAMP elevation, two early events during the lipolysis response to TNF-alpha. Furthermore, salicylate prevents the down regulation of cyclic-nucleotide phosphodiesterase 3B, an enzyme responsible for cAMP hydrolysis. Perilipins coat intracellular lipid droplet surface by restricting lipase access to the triacylglycerol substrates. TNF-alpha down regulates perilipin but promotes its phosphorylation during lipolysis stimulation; these actions are efficiently reversed by salicylate. Salicylate slightly reduces basal but completely inhibits TNF-alpha-liberated lipase activity. In contrast, neither salicylate nor TNF-alpha alters the protein levels of hormone-sensitive lipase and adipose triglyceride lipase. In addition, sodium salicylate restricts basal lipolysis simulated by a high concentration of glucose and significantly diminishes the high glucose-enhanced lipolysis response to TNF alpha. These results provide novel evidence that salicylate directly blocks TNF alpha-mediated FFA efflux from adipocytes, hence reducing plasma FFA levels and increasing insulin sensitivity. PMID- 17911534 TI - FP prostanoid receptor-mediated induction of the expression of early growth response factor-1 by activation of a Ras/Raf/mitogen-activated protein kinase signaling cascade. AB - FP prostanoid receptors are G-protein-coupled receptors whose physiological activator is prostaglandin-F(2alpha) (PGF(2alpha)). PGF(2alpha) has been implicated in wound healing and cardiac hypertrophy, which are both known to involve the induction of the immediate-early response gene, early growth response factor-1 (EGR-1). We hypothesized that activation of the human FP receptor by PGF(2alpha) could induce the expression of EGR-1 and found that 1 muM PGF(2alpha) produced a time-dependent induction of both mRNA and protein expression for EGR 1. This FP receptor-mediated induction of EGR-1 expression involved activation of the small GTPase Ras followed by activation of C-Raf and the mitogen-activated protein (MAP) kinase kinases 1 and 2 (MEK1/2). Thus, induction of EGR-1 expression by PGF(2alpha) was blocked using dominant-negative constructs of Ras and C-Raf and the Raf kinase inhibitor 4-(4-(3-(4-chloro-3 trifluoromethylphenyl)ureido)phenoxy)-pyridine-2-carboxyllic acid methyamide-4 methylbenzenesulfonate (BAY43-9006). Likewise, the MEK1/2 inhibitor 2'-amino-3' methoxyflavone (PD98059) blocked the induction of EGR-1 expression by PGF(2alpha). FP receptor stimulation by PGF(2alpha) induced the phosphorylation of C-Raf, MEK1/2, and extracellular signal-regulated kinases 1 and 2, consistent with the activation of a MAP kinase signaling cascade. PGF(2alpha) was also found to induce the expression of EGR-1 in rat cardiomyocytes through the activation of endogenous FP receptors. This induction of EGR-1 expression in cardiomyocytes also involved the activation of Raf and MAP kinase signaling and was dependent on the activation of protein kinase C. This is the first report to show the regulation of EGR-1 expression after PGF(2alpha) activation of FP receptors and suggests that this could be an early event involved in wound healing and cardiac hypertrophy. PMID- 17911535 TI - Patient-specific time to peak abdominal organ enhancement varies with time to peak aortic enhancement at MR imaging. AB - PURPOSE: To retrospectively evaluate the relationship between the times to peak enhancement of the liver, pancreas, and jejunum with respect to the time to peak aortic enhancement at magnetic resonance (MR) imaging. MATERIALS AND METHODS: The committee on human research approved this study and waived written informed consent. This study was HIPAA compliant. The study retrospectively identified 141 patients (63 men, 78 women; mean age, 57 years) who underwent abdominal MR imaging by using a test bolus that was monitored approximately every second for 2 minutes with a spoiled gradient-echo T1 transverse section through the upper abdomen. The times to peak enhancement of the aorta, liver, pancreas, and jejunum were recorded and correlated with the time to peak aortic enhancement, age, and sex by means of univariate and multivariate linear regression analyses. RESULTS: The mean time to peak aortic enhancement was 21.1 seconds (range, 8.7-41.8 seconds). The times to peak enhancement of the liver, pancreas, and jejunum were positively and linearly correlated with the time to peak aortic enhancement (r = 0.69, 0.86, and 0.80, respectively, all P < .001) and were 3.39, 1.64, and 2.04 times longer than the time to peak aortic enhancement, respectively. Age, sex, and history of heart disease did not give additional predictive information for determining the time to peak visceral enhancement. CONCLUSION: The times to peak enhancement of the liver, pancreas, and jejunum are linearly related to that of the aorta. These results could potentially allow tailored patient- and organ specific scan delay optimization at contrast material-enhanced MR image evaluation. PMID- 17911536 TI - Minor head injury: guidelines for the use of CT--a multicenter validation study. AB - PURPOSE: To prospectively and externally validate published national and international guidelines for the indications of computed tomography (CT) in patients with a minor head injury. MATERIALS AND METHODS: The study protocol was institutional review board approved. All patients implicitly consented to use of their deidentified data for research purposes. Between February 2002 and August 2004, data were collected in consecutive adult patients with blunt minor head injury (Glasgow Coma Scale score of 13-14 or 15) and a risk factor for neurocranial traumatic complications at presentation at four Dutch university hospitals. Primary outcome was any neurocranial traumatic CT finding. Secondary outcomes were clinically relevant traumatic CT findings and neurosurgical intervention. Sensitivity and specificity of each guideline for all outcomes and the number of patients needed to scan to detect one outcome (ie, the number of patients needed to undergo CT to find one patient with a neurocranial traumatic CT finding, a clinically relevant traumatic CT finding, or a CT finding that required neurosurgical intervention) were estimated. RESULTS: Data were available for 3181 patients. Only the European Federation of Neurological Societies guidelines reached a sensitivity of 100% for all outcomes. Specificity was 0.0% 0.5%. The Dutch guidelines had the lowest sensitivity (76.5%) for neurosurgical interventions. The best specificities for traumatic CT findings and neurosurgical interventions were reached with the criteria proposed by the United Kingdom National Institute for Clinical Excellence (NICE) (46.1% and 43.6%, respectively), albeit at relatively low sensitivities (82.1% and 94.1%, respectively). The number of patients needed to scan ranged from six to 13 for traumatic CT findings and from 79 to 193 for neurosurgical interventions. CONCLUSION: All validated guidelines demonstrated a trade-off between sensitivity and specificity. The lowest number of patients needed to scan for either of the outcomes was reached with the NICE criteria. SUPPLEMENTAL MATERIAL: radiology.rsnajnls.org/cgi/content/full/2452061509/DC1 (c) RSNA, 2007. PMID- 17911538 TI - Breast lymphoma: imaging findings of 32 tumors in 27 patients. AB - PURPOSE: To retrospectively evaluate the imaging findings of breast lymphomas in patients who had undergone mammography, ultrasonography (US), magnetic resonance (MR) imaging, or combined positron emission tomography (PET)/computed tomography (CT) scanning. MATERIALS AND METHODS: The institutional review board approved this HIPAA-compliant study and waived informed consent. Twenty-seven women who had been diagnosed with breast lymphoma (32 tumors) and had undergone preoperative imaging were identified from the surgical pathology database (mean age, 51 years; median, 55 years; range, 19-78 years at time of diagnosis). Two radiologists reviewed the mammographic, US, and MR images. One nuclear medicine physician reviewed the PET/CT scans. All available pathologic specimens were reviewed by a hematologic pathologist. RESULTS: The mean tumor size at diagnosis was 2.9 cm (range, 1-5 cm). Seventeen tumors manifested with a palpable mass, two with diffuse enlargement of the breast, and 13 were asymptomatic. Twenty-two women underwent mammography; 24, US; one, MR imaging; and 10, PET/CT scanning. Mammograms of 25 tumors showed a noncalcified mass in 19, global asymmetry in four, focal asymmetry in one, and no abnormality in one. US of 29 tumors showed a mass in 26 and diffuse architectural distortion in three. Masses typically were irregular, hypoechoic, and hypervascular and demonstrated indistinct margins or an echogenic boundary. Dynamic contrast material-enhanced MR imaging of one tumor showed an intensely and heterogeneously enhancing mass with rapid enhancement and washout characteristics. PET/CT scans of 13 tumors showed intense diffuse hypermetabolism in 12 and response to therapy in all 12 tumors. CONCLUSION: The imaging findings reported in this study should alert the radiologist to a possible diagnosis of breast lymphoma. PMID- 17911537 TI - Enhanced infarct border zone function and altered mechanical activation predict inducibility of monomorphic ventricular tachycardia in patients with ischemic cardiomyopathy. AB - PURPOSE: To prospectively determine whether mechanical behavior of left ventricular wall segments that contain different degrees of scar tissue and are located at different distances from the interface between infarcted and noninfarcted myocardial tissue can help predict inducibility of monomorphic ventricular tachycardia (VT) in patients with ischemic cardiomyopathy. MATERIALS AND METHODS: This HIPAA-compliant study was institutional review board approved; written informed consent was obtained from all patients. Forty-six patients (36 men, 10 women; mean age +/- standard deviation, 61.6 years +/- 11.9) with prior myocardial infarction (MI) and left ventricular dysfunction were referred for defibrillator implantation and underwent an electrophysiologic examination and tagged contrast-enhanced magnetic resonance (MR) imaging. Peak circumferential shortening strain (Ecc) and time to peak Ecc were measured in 12 segments from short-axis sections. Remote, adjacent, and border zones were defined according to increasing proximity to the MI. Patients in whom monomorphic VT could be induced (ie, inducible patients) were considered positive for inducibility. Relationships between inducibility of monomorphic VT, peak Ecc, and time to peak Ecc were analyzed with one-way analysis of variance and Bonferroni test. RESULTS: Inducible patients had more infarcted and border zone sectors and a shorter time to peak Ecc than did noninducible patients in the border zone and adjacent and infarcted regions (P < .001). Peak Ecc in the border zone of inducible patients ( 11.42% +/- 0.46 [standard error]) was greater than that in noninducible patients (-10.18% +/- 0.38; P < .05). Ratio of Ecc in border zone and in remote regions was greater (P < .05) in inducible patients than in noninducible patients (1.31 +/- 0.27 vs 0.64 +/- 0.13, respectively). CONCLUSION: Enhanced border zone function defined as greater Ecc and earlier time to peak Ecc showed positive correlation to VT inducibility in patients with prior MI and left ventricular dysfunction. PMID- 17911539 TI - Malignant cervical lymphadenopathy: diagnostic accuracy of diffusion-weighted MR imaging. AB - PURPOSE: To prospectively determine the diagnostic accuracy of diffusion-weighted magnetic resonance (MR) imaging for discrimination of malignant neck nodes due to lymphoma, squamous cell carcinoma (SCC), and undifferentiated nasopharyngeal carcinoma (NPC), with histologic findings and imaging criteria as reference standards. MATERIALS AND METHODS: Ethics committee approval and informed consent were obtained. Patients with malignant lymphadenopathy underwent 1.5-T diffusion weighted MR imaging. A region of interest was drawn around the malignant node on apparent diffusion coefficient (ADC) maps; ADC values were compared (Kruskal Wallis test). Receiver operating characteristic analysis was employed to investigate whether ADC values could aid in discrimination among malignancies. RESULTS: Forty-three patients (34 men, nine women; mean age, 54 years) with 43 nodes underwent imaging. Mean ADC values for lymphoma (n = 8), NPC (n = 17), and SCC (n = 18) were (0.664 +/- 0.071 [standard deviation]) x 10(-3) mm(2)/sec, (0.802 +/- 0.128) x 10(-3) mm(2)/sec, and (1.057 +/- 0.169) x 10(-3) mm(2)/sec, respectively, with significant differences between SCC and lymphoma or NPC (P < .001) and between NPC and lymphoma (P = .04). To optimize sensitivity and specificity with equal weighting, ADC threshold values for distinguishing between SCC and NPC, between SCC and lymphoma, and between NPC and lymphoma were 0.894 x 10(-3) mm(2)/sec, 0.824 x 10(-3) mm(2)/sec, and 0.694 x 10(-3) mm(2)/sec, respectively. To produce a 100% specificity while sensitivity is maximized, the following ADC threshold values were obtained for prediction of differentiation between malignancies: (a) SCC versus lymphoma, greater than 0.824 x 10(-3) mm(2)/sec (sensitivity, 94%), and lymphoma versus SCC, less than 0.767 x 10(-3) mm(2)/sec (sensitivity 88%); (b) NPC versus SCC, less than 0.764 x 10(-3) mm(2)/sec (sensitivity, 47%), and SCC versus NPC, greater than 1.093 x 10(-3) mm(2)/sec (sensitivity, 39%); (c) NPC versus lymphoma, greater than 0.788 x 10( 3) mm(2)/sec (sensitivity, 53%), and lymphoma versus NPC, no suitable threshold value. CONCLUSION: Diffusion-weighted MR imaging shows significant differences among malignant nodes of SCC, lymphoma, and NPC. ADC threshold values can help distinguish SCC from lymphoma. SUPPLEMENTAL MATERIAL: http://radiology.rsnajnls.org/cgi/content/full/2451061804/DC1. PMID- 17911541 TI - Randomized clinical trials: impact on clinical practice for symptomatic and asymptomatic extracranial carotid occlusive disease. AB - Carotid artery stenting (CAS) has emerged as a useful and potentially less invasive alternative to carotid endarterectomy (CEA) for the treatment of extracranial carotid stenoses. In this regard, it has been suggested that specific patient subgroups who may benefit from CAS including those with significant medical comorbidities, recurrent stenosis, anatomically inaccessible lesions, and a hostile neck. However, the purpose of this report is to evaluate whether or not CAS should replace CEA in the treatment of symptomatic and asymptomatic disease in better risk patients, also. This report contains results from recently published randomized clinical trials. PMID- 17911542 TI - Commentary on "Randomized clinical trials: impact on clinical practice for symptomatic and asymptomatic extracranial carotid occlusive disease". PMID- 17911543 TI - Carotid artery stenting will replace carotid endarterectomy. AB - Stroke is the third leading cause of death in the United States. Carotid artery stenosis represents one of the most common etiologies of stroke. The current treatment modalities available for the treatment of carotid artery stenosis are carotid endarterectomy (CEA) and carotid artery stenting (CAS). Several clinical trials comparing CEA with medical management showed superiority of the surgical arm; however, the applicability of these results to the general population is limited by the fact that the patients and surgeons enrolled in these trials were carefully selected, and the optimal medical therapy used does not meet the current treatment standards. Carotid artery stenting has emerged as a treatment alternative to CEA, as shown in randomized trials comparing the 2 treatment modalities. Recent data from large-volume CAS registries indicate that percutaneous treatment of carotid artery stenosis compares favorably to CEA. Furthermore, the CAS trial designs make these results more applicable to the community standards. These data suggest that CAS will become the treatment of choice in patients with carotid artery stenosis. PMID- 17911544 TI - Commentary on "Carotid artery stenting will replace carotid endarterectomy". PMID- 17911545 TI - Endovascular techniques for supra-aortic trunk intervention. AB - Endovascular techniques have gradually become the preferred treatment modality for occlusive disease of the supra-aortic trunks (SAT). Stenotic lesions are particularly amenable to catheter-based therapy. Standard cervical bypasses or intrathoracic reconstruction is now generally reserved for cases of extensive, multivessel involvement or after failure of previous endovascular procedures. Angiographic assessment of aortic arch anatomy and equipment and technical choices for arch branch catheterization constitute the foundations. The left subclavian artery is most frequently diseased; interventional options revolve around antegrade and retrograde techniques. Preservation of the left vertebral artery-and at times of the internal mammary-is an important issue. Treatment of lesions in the proximal common carotid arteries tends to be straightforward when dealing with focal stenoses. Embolic protection should be used whenever feasible and reasonable. The innominate is often the most challenging SAT vessel because of its large diameter and short length, as well as the fact that it bifurcates into the right subclavian and common carotid arteries. Right subclavian artery intervention tends to be more difficult than its left-side counterpart, which is related to fluoroscopic visualization and the tendency for stenoses to develop in the very short segment between its origin and the take-off of the right vertebral artery. Supra-aortic trunk intervention has become more commonplace at present, but it is frequently difficult and carries significant potential for failure and mishap. These risks can be minimized through a complete understanding of the full spectrum of technical and catheter choices that are available to the interventionist. PMID- 17911546 TI - Commentary on "Endovascular techniques for supra-aortic trunk intervention". PMID- 17911547 TI - Should statins be given routinely before carotid endarterectomy? AB - Statins belong to a class of drugs known to inhibit 3-hydroxy 3-methylglutaryl coenzyme A (HMG CoA) reductase, and block hepatic cholesterol synthesis. Statins have been found to be highly effective in primary and secondary stroke prevention among medically managed patients with cardiovascular disease, and it appears that this benefit is largely owing to the non-cholesterol-lowering, so called pleiotropic, effects of statins. Over the past decade, agents such as beta blockers, aspirin, or other antiplatelet medications have proven to reduce the incidence of adverse postoperative outcomes among vascular surgical patients and have rightfully assumed a place in our overall therapeutic armamentarium. There is growing evidence that statins may be especially effective in reducing cardiovascular morbidity and improving outcome following major vascular surgery. A recent study from Johns Hopkins Hospital demonstrated a threefold reduction in the rate of perioperative stroke (P < .05) and fivefold reduction of perioperative mortality (P < .05) among 1566 patients undergoing carotid endarterectomy (CEA). This benefit was confirmed in a series of 3360 CEAs performed at multiple hospitals throughout western Canada. Statin use was independently associated with a 75% reduction (OR: 0.25; 95%CI: 0.07-0.90) in the odds of death and a 45% reduction (OR: 0.55; 95% CI: 0.32-0.95) in the odds of ischemic stroke or death among patients with symptomatic carotid disease. A number of the pleiotropic effects of statin medications may be responsible for these clinical observations. Further work is necessary to better elucidate these mechanisms, as well as to determine the optimal agents, dosing, and timing of drug administration among patients undergoing carotid interventions. Nevertheless, in light of these data a strong case can be made to start patients on statin medications prior to CEA if time permits. PMID- 17911548 TI - Commentary on "Should statins be given routinely before carotid endarterectomy?". PMID- 17911549 TI - Contemporary management of carotid stenosis: carotid endarterectomy is here to stay. AB - Thromboembolic complications from atherosclerotic disease of the carotid bifurcation are a common etiology of ischemic stroke, which has a significant morbidity and mortality. Carotid artery angioplasty and stenting (CAS) has been proposed as an alternative therapy for patients requiring treatment of carotid artery stenosis as opposed to the "gold standard" surgical intervention, carotid endarterectomy (CEA). Intense debate regarding these 2 therapeutic approaches has centered on their respective safety profiles. To date, despite multiple studies, no convincing evidence demonstrates the superiority or even equality of CAS to the proven safety, efficacy, and durability of CEA. This chapter reviews in detail the available evidence for CAS and CEA and provides a rationale that, given the available data, CEA remains the preferred therapy for the majority of patients who require treatment of carotid artery stenosis. PMID- 17911550 TI - Commentary on "Contemporary management of carotid stenosis: carotid endarterectomy is here to stay". PMID- 17911551 TI - Treatment of celiac artery compression syndrome: does it really exist? AB - Celiac artery syndrome exists, although it remains controversial, and in some patients a firm diagnosis cannot be established. Duplex scanning or computed tomographic, magnetic resonance, or contrast aortography will confirm intermittent or permanent compression of the celiac artery by the crus of the diaphragm, the median arcuate ligament, or fibrous ganglionic tissue. Preoperative ganglion block and exercise gastric tonometry are useful diagnostic tools to predict better outcome after treatment. In patients with well-defined syndrome of chronic mesenteric and gastric ischemia or with exercise-induced pain, good results can be expected with division of the median arcuate ligament with open celiac artery reconstruction. Patients with atypical pain or history of psychiatric disorders only occasionally benefit from surgical repair. The role of primary stenting of celiac artery compression is still not well defined, and current data do not support the use of balloon-expandable stents. Laparoscopic division of the median arcuate ligament followed by celiac artery stenting is an effective, minimally invasive technique to manage selected patients with celiac artery compression syndrome. PMID- 17911552 TI - Commentary on "Treatment of celiac artery compression syndrome: does it really exist?". PMID- 17911553 TI - The role of embolic protection devices in renal artery stenting. AB - Renal artery angioplasty and stenting is commonly performed for the treatment of hypertension and ischemic nephropathy. An increasing number of procedures are being undertaken for "renal preservation" despite an associated risk of renal function decline related to the embolization of atheromatous debris liberated during the procedure. Although smaller, more flexible guidewires and stents have been developed to decrease the amount of debris created, interest in the off label use of embolic protection devices has increased. We review the available embolic protection devices and currently available data regarding their use in renal artery interventions. Although not designed for use in the renal artery, there are at least theoretical reasons to believe that embolic protection during renal artery angioplasty may improve outcomes. PMID- 17911554 TI - Commentary on "The role of embolic protection devices in renal artery stenting". PMID- 17911555 TI - Endovascular aortic repair should be the gold standard for ruptured AAAs, and all vascular surgeons should be prepared to perform them. AB - When ruptured abdominal aortic aneurysms (AAAs) are not treated, they cause death. In addition, ruptured abdominal aortic aneurysms (RAAAs) have high mortality (35%-70%) and morbidity rates when treated by standard open surgical methods. These high perioperative mortality and morbidity rates have not been substantially reduced despite the introduction of many improvements in open operative technique or perioperative care. Endovascular approaches to treat AAAs introduced in the early 1990s provided an opportunity to substantially alter treatment outcomes when rupture occurred. This article details how these endovascular approaches, which include endovascular stented grafts, can be applied to the treatment of RAAAs, and what advantages these new catheter-based approaches to treatment offer. PMID- 17911556 TI - Commentary on "Endovascular aortic repair should be the gold standard for ruptured AAAs and all vascular surgeons should be prepared to perform them". PMID- 17911557 TI - New techniques and developments to treat long infrainguinal arterial occlusions: use of reentry devices, subintimal angioplasty, and endografts. AB - The endovascular recanalization of long infrainguinal arterial occlusions has made significant progress in the past decade. The technique of subintimal angioplasty has opened the door to the treatment of lesions uncross-able using standard transluminal approaches. With the advent of new wires and catheters designed to traverse long lesions, and reentry devices used to facilitate the subintimal approach, percutaneous treatment has made substantial inroads into territory previously dominated by surgical bypass. Advances in stent technology have improved deliverability and patency in these difficult applications. Percutaneously delivered covered endografts into the femoropopliteal segment may allow better patency by performing like endoluminal prosthetic bypasses. In this article, we review the latest technology available to treat occlusions of the femoropopliteal arterial segment. PMID- 17911558 TI - Commentary on "New techniques and developments to treat long infrainguinal arterial occlusions: use of reentry devices, subintimal angioplasty, and endografts. PMID- 17911559 TI - Biological treatment of vein grafts and stents in lower-extremity arterial reconstruction. AB - Longevity of lower-extremity revascularization procedures has typically been limited because of restenosis of grafts, as well as stents and other percutaneous techniques. The ability to prevent or ameliorate neointimal hyperplasia and improve durability of lower-extremity arterial reconstruction is the focus of significant scientific and clinical research. The transcription factor decoy edifoligide was investigated as a potential inhibitor of neointimal hyperplasia, but the results of the pivotal clinical trial did not demonstrate significant improvement in graft reintervention, although secondary patency was improved. Similar to the coronary circulation, drug-eluting stents are a potential tool for prevention of restenosis after percutaneous arterial reconstruction. The SIROCCO study did not demonstrate improvement in superficial femoral artery stent patency with sirolimus-eluting stents. Studies are underway that are investigating paclitaxel-eluting stents for use in the superficial femoral artery. Other potential mediators of restenosis include absorbable drug-eluting stents and antibody-coated stents designed to promote endothelialization of the stent or graft surface. In addition, absorbable wraps eluting paclitaxel can be used at arterial and arteriovenous anastomoses to prevent restenosis. Clinical trials investigating these novel technologies are underway. PMID- 17911560 TI - Commentary on "Biological treatment of vein grafts and stents in lower-extremity arterial reconstruction". PMID- 17911561 TI - New techniques and developments of stenting for infrainguinal arterial occlusive disease: are the results any better than balloon angioplasty alone? AB - Infrainguinal occlusive disease is a complex problem necessitating the cooperation of both medical and surgical therapies to aid limb salvage and alleviate symptoms. Endovascular therapies are varied, with no treatment clearly outweighing the other in terms of efficacy and durability. Angioplasty for focal stenosis has gained ground as the treatment of choice when indicated. There has also been a rapid evolution in stent technology, from early stainless steel wall stents to today's drug eluting nitinol stents. In this article, we examine the literature on these new technologies and treatment options and make recommendations based on the best data available. PMID- 17911562 TI - Commentary on "New techniques and developments of stenting for infrainguinal arterial occlusive disease: are the results any better than balloon angioplasty alone?". PMID- 17911563 TI - Radiofrequency ablation of the great saphenous vein performed in the office: tips for better patient convenience and comfort and how to perform it in less than an hour. AB - Radiofrequency ablation of the great saphenous vein has become increasingly popular for its decreased morbidity and greater patient satisfaction as compared with conventional saphenous vein stripping. Because of the relative simplicity and safety of this procedure and fueled by changes in reimbursement, endovenous radiofrequency ablation is now most often performed as an office-based procedure. This sets the stage for developing new technology that can achieve better results in a shorter time without compromise to patient safety and satisfaction. The balance between patient satisfaction and increased productivity is tenuous, and the former should never be sacrificed for the latter. The recipe for such a seemingly formidable task begins the moment the patient sets foot in the waiting area. Recognizing the steps that cause patient anxiety and strategically turning them into comfort zones takes a concerted effort between the surgeon and office staff. Here, we demonstrate how simple changes can transform an office-based endovenous practice into one in which patient and physician alike are maximally satisfied. PMID- 17911564 TI - Commentary on "Radiofrequency ablation of the great saphenous vein performed in the office: tips for better patient convenience and comfort and how to perform it in less than an hour". PMID- 17911565 TI - The economics of vein disease. AB - The management of cosmetic vein problems requires a very different approach than that for the majority of most other vascular disorders that occur in a vascular surgery practice. This article focuses on the business aspects of a cosmetic vein practice, with particular attention to the uniqueness of these issues. Managing patient expectations is critical to the success of a cosmetic vein practice. Maneuvering within the insurance can be difficult and frustrating for both the patient and the practice. Practices should use cost accounting principles to evaluate the success of their vein work. Vein surgery--especially if performed within the office--can undergo an accurate break-even analysis to determine its profitability. PMID- 17911566 TI - Commentary on "The economics of vein disease". PMID- 17911567 TI - The new anticoagulants. AB - Currently, the anticoagulants available are limited to warfarin, heparin compounds, and direct thrombin inhibitors. Warfarin, the most commonly used outpatient anticoagulant, has significant shortcomings. There are numerous drug/drug and drug/food interactions, and there is difficulty in dosing for one third of patients. Numerous new anticoagulants are undergoing testing. The meta pentasaccharides use the core molecule of heparin and replace one or more of the sulphated groups. They are administered by subcutaneous injection, but they require only once per week dosing. Numerous studies have shown equivalents or superiority to warfarin in treatment for thromboembolic events. Direct thrombin inhibitors block thrombin (IIa) and are used mostly for heparin-induced thrombosis or during coronary interventions. However, there is an orally administered direct thrombin inhibitor, Dabigatran, which is undergoing phase III testing. It can be given without regard to weight, age, or gender with minimal drug interactions. This drug has been proven to be equivalent to low molecular weight heparin (LMWH) in deep venous thrombosis (DVT) prophylaxis and showed no excess bleeding. Another promising group of anticoagulants with numerous investigations underway are the direct X inhibitors. Most are excreted renally as opposed to hepatic clearance with intravenous or oral dosing. Numerous phase II studies have shown them to have equivalent or superior prophylaxis for DVT when compared with LMWH. Oral IXa inhibitors and an oral thrombin cofactor inhibitor are also under development. It is clear that in the near future, anticoagulants will be available that offer significant advantages when compared to those currently in use. PMID- 17911568 TI - Commentary on "The new anticoagulants". PMID- 17911569 TI - Commentary. Comparison of fixed-dose weight-adjusted unfractionated heparin and low-molecular-weight heparin for acute treatment of venous thromboembolism. PMID- 17911570 TI - Commentary. Risk score for peri-interventional complications of carotid artery stenting. PMID- 17911571 TI - Commentary. Transition from primary Raynaud's phenomenon to secondary Raynaud's phenomenon identified by diagnosis of an associated disease. Results of ten years of prospective surveillance. PMID- 17911572 TI - Commentary. Angiotensin converting enzyme inhibitors and aortic rupture: population based case control study. PMID- 17911573 TI - Gender differences in Chinese immigrants: predictors for antihypertensive medication adherence. AB - The purpose of this study was to describe gender differences in predictors for antihypertensive medication adherence in Chinese immigrants. A cross-sectional design was used with recruitment with a convenience sample of 100 men and 100 women with hypertension. Measurements for demographics, cultural factors, clinical factors, and medication adherence were self-administered by the participants. Blood pressure was checked twice. A multivariate logistic regression was used to establish a parsimonious prediction model for medication adherence. It was found that in men, longer length of stay in the United States was a predictor for nonadherence. The predictor of nonadherence in women was lower perceived benefits of antihypertensive medications. To increase adherence in women, the benefits of antihypertensive medications should be emphasized. For men who have lived in the United States for 12 years or more, their adherence should be closely monitored. PMID- 17911574 TI - A comparison of the use of home care services by Anglo-American and Mexican American elders. AB - Elders' use of home care services varies globally. The authors tested the relationship of cultural/ethnic, contextual, and social structural factors with elders' use of home care services in the United States. Fifty-seven non-Hispanic White or "Anglo" and 56 Mexican American elders participated. Use of services significantly correlated with Anglo and Mexican American elders' cultural/ethnic and contextual factors. Perceived need and service awareness were the two factors that were significant for both groups and that the authors believe would be amenable to change in future transcultural nursing interventions to decrease use of services disparities. PMID- 17911575 TI - Participatory action research (PAR): an approach for improving black women's health in rural and remote communities. AB - Women are among the most disadvantaged members of any community, and they tend to be at greatest risk of illness. Black women are particularly vulnerable and more prone than White women to illnesses associated with social and economic deprivation, including heart disease and diabetes. They utilize preventive health services less often, and when they fall ill, the health of their families and communities typically suffers as well. This article discusses the process of doing innovative participatory action research (PAR) in southwest Nova Scotia Black communities. The effort resulted in the generation of a database, community action, and interdisciplinary analysis of the intersecting inequities that compromise the health and health care of African Canadian women, their families, and their communities. This particular research effort serves as a case study for explicating the key tenets of PAR and the barriers to and contradictions in implementing PAR in a community-academic collaborative research project. PMID- 17911576 TI - How much should we expect? Family caregiving of AIDS patients in rural Uganda. AB - The aim of this study was to measure the burden of care for family caregivers of AIDS patients. A cross-sectional exploratory design was used to describe the care experiences of family caregivers of AIDS care recipients. A questionnaire was used to interview 120 family caregivers of AIDS patients from four rural areas in western Uganda. The questions asked were related to 12 domains of family caregiving. Care burden scores of caregivers were calculated. It was found that care burden scores were high in all domains except those regarding relationships within the families and substance abuse. Serious work overload and low health status were reported. The high burden of caregiving puts family caregivers at risk for decreased health status and increased social isolation and depression. PMID- 17911577 TI - Addressing childhood overweight: strategies learned from one Latino community. AB - The purpose of this study is to understand one Latino community's perspective about childhood overweight within this high-risk ethnic group. Three focus groups, consisting of 12 mothers, 12 fathers, and 8 boys and 4 girls ages 10-12, participated. Transcripts of interviews were coded using N-VIVO and analyzed thematically. Several themes emerge: parents' demanding work schedules, lack of time, transportation issues, opportunities for physical activities, and lack of meal preparation. Participants knew good nutrition and exercise help prevent obesity. Nursing interventions must address multiple challenges with childhood obesity at the family and community levels. PMID- 17911579 TI - Cutting edge: IL-4-mediated protection of primary B lymphocytes from apoptosis via Stat6-dependent regulation of glycolytic metabolism. AB - IL-4 prevents the death of naive B lymphocytes through the up-regulation of antiapoptotic proteins such as Bcl-x(L). Despite studies implicating glucose utilization in growth factor-dependent survival of hemopoietic cells, the role of glucose energy metabolism in maintaining B cell viability by IL-4 is unknown. We show that IL-4 triggers glucose uptake, Glut1 expression, and glycolysis in splenic B cells; this is accompanied by increased cellular ATP. Glycolysis inhibition results in apoptosis, even in the presence of IL-4. IL-4-induced glycolysis occurs normally in B cells deficient in insulin receptor substrate-2 or the p85alpha subunit of PI3K and is not affected by pretreatment with PI3K or MAPK pathway inhibitors. Stat6-deficient B cells exhibit impaired IL-4-induced glycolysis. Cell-permeable, constitutively active Stat6 is effective in restoring IL-4-induced glycolysis in Stat6-deficient B cells. Therefore, besides controlling antiapoptotic proteins, IL-4 mediates B cell survival by regulating glucose energy metabolism via a Stat6-dependent pathway. PMID- 17911580 TI - Functional human antigen-specific T cells produced in vitro using retroviral T cell receptor transfer into hematopoietic progenitors. AB - In vitro production of human T cells with known Ag specificity is of major clinical interest for immunotherapy against tumors and infections. We have performed TCRalphabeta gene transfer into human hemopoietic progenitors from postnatal thymus or umbilical cord blood, and subsequently cultured these precursors on OP9 stromal cells expressing the Notch human ligand Delta-like1. We report here that fully mature, functional T cells with controlled Ag specificity are obtained from such cultures. Using vectors encoding TCRalphabeta-chains directed against melanoma (MART-1), viral (CMV), and minor histocompatibility (HA 2) Ags, we show that the obtained Ag-specific T cells exert cytolytic activity against their cognate Ag and expand in vitro upon specific TCR stimulation. Therapeutic applications may arise from these results because they provide a way to produce large numbers of autologous mature Ag-specific T cells in vitro from undifferentiated hemopoietic progenitors. PMID- 17911581 TI - Regulatory T cells prevent CD8 T cell maturation by inhibiting CD4 Th cells at tumor sites. AB - Natural regulatory T cells (Tregs) are present in high frequencies among tumor infiltrating lymphocytes and in draining lymph nodes, supposedly facilitating tumor development. To investigate their role in controlling local immune responses, we analyzed intratumoral T cell accumulation and function in the presence or absence of Tregs. Tumors that grew in normal BALB/c mice injected with the 4T1 tumor cell line were highly infiltrated by Tregs, CD4 and CD8 cells, all having unique characteristics. Most infiltrating Tregs expressed low levels of CD25Rs and Foxp3. They did not proliferate even in the presence of IL-2 but maintained a strong suppressor activity. CD4 T cells were profoundly anergic and CD8 T cell proliferation and cytotoxicity were severely impaired. Depletion of Tregs modified the characteristics of tumor infiltrates. Tumors were initially invaded by activated CD4(+)CD25(-) T cells, which produced IL-2 and IFN-gamma. This was followed by the recruitment of highly cytotoxic CD8(+) T cells at tumor sites leading to tumor rejection. The beneficial effect of Treg depletion in tumor regression was abrogated when CD4 helper cells were also depleted. These findings indicate that the massive infiltration of tumors by Tregs prevents the development of a successful helper response. The Tregs in our model prevent Th cell activation and subsequent development of efficient CD8 T cell activity required for the control of tumor growth. PMID- 17911582 TI - The role of apoptosis in the ameliorating effects of a CDR1-based peptide on lupus manifestations in a mouse model. AB - Experimental systemic lupus erythematosus (SLE) can be induced in mice following immunization with an anti-DNA mAb expressing a major Id, 16/6Id. Treatment with a peptide, designated human CDR1 (hCDR1; Edratide), that is based on the sequence of CDR1 of the 16/6Id ameliorated disease manifestations. In the present study, we investigated the roles of apoptosis and related molecules in BALB/c mice with induced experimental SLE following treatment with hCDR1. A higher state of activation and increased rate of apoptosis were found in lymphocytes of SLE afflicted mice as compared with healthy controls. The latter effects were associated with up-regulated caspase-8 and caspase-3, and down-regulated Bcl x(L). The ameliorative effects of hCDR1 were associated with down-regulation of caspase-8 and caspase-3, up-regulation of Bcl-x(L), and a reduced rate of apoptosis. Treatment of diseased mice with an apoptosis-reducing compound that inhibited caspases down-regulated the secretion of the pathogenic cytokine IFN gamma and lowered the intensity of glomerular immune complex deposits and the levels of proteinuria. Furthermore, coincubation of Bcl-x(L) inhibitors with hCDR1-treated cells abrogated the ability of hCDR1 to reduce the activation state of lymphocytes and to down-regulate the secretion of IL-10 and IFN-gamma. Moreover, the Bcl-x(L)-expressing CD4(+)CD25(+) cells from hCDR1-treated mice induced the expression of Bcl-x(L) in CFSE-labeled CD4(+)CD25(-) cells of the SLE afflicted mice. Thus, the reduction of apoptosis and the up-regulation of Bcl x(L), which plays an apparent role in tolerance induction, contribute to at least part of the beneficial effects of hCDR1 on lupus manifestations. PMID- 17911584 TI - Maintenance of surrogate light chain expression induces developmental delay in early B cell compartment. AB - The production of a mature B cell requires passage through a number of developmental checkpoints. The pre-BCR plays a critical role in passage through the pro-B cell/pre-B cell checkpoint, and thus plays a central role in regulating the differentiation of a B cell. Due to the significance of this receptor, it is imperative that pre-BCR expression and function are precisely regulated. In this study, we have investigated a system in which the regulation of the pre-BCR is altered. We have found that continued expression of components of the pre-BCR (lambda5) resulted in a delay in the kinetics of B cell maturation. Pro-B cells from normal mouse bone marrow retrovirally infected with lambda5 exhibited a delay in differentiation. As compared with wild-type cells at the same time point, there is a reduction in the presence of cell surface markers that indicate developmental progression, and there is a 6- to 16-fold decrease in the production of Ig-positive cells in B cell maturation assays. The capacity to alter B cell progression by modifying and extending pre-BCR expression argues that the receptor and its associated signals play a unique role in directing developmental outcomes. PMID- 17911583 TI - The surprising kinetics of the T cell response to live antigenic cells. AB - Cooperation between CD4(+) and CD8(+) T cells is required for the proper development of primary effector and memory CD8(+) T cells following immunization with noninflammatory immunogens. In this study, we characterized murine CD4(+) and CD8(+) T cell responses to male-specific minor histocompatibility (HY) Ags following injection of live male cells into females of the same strain. Male cells are rejected 10-12 days after transfer, coinciding with the expansion and effector function of CD8(+) CTLs to two H-2D(b)-restricted epitopes. Although anti-HY CD4(+) T cell responses are readily detectable day 5 posttransfer, CD8(+) responses are undetectable until day 10. The early CD4(+) response is not dependent on direct presentation of Ag by donor male cells, but depends on presentation of the male cells by recipient APC. The CD4(+) T cell response is required for the priming of CD8(+) T cell effector responses and rejection of HY incompatible cells. Unexpectedly, HY-specific CD4(+) T cells are also capable of efficiently lysing target cells in vivo. The delay in the CD8(+) T cell response can be largely abrogated by depleting T cells from the male inoculum, and donor male CD8(+) T cells in particular suppress host anti-HY CD8(+) responses. These data demonstrate dramatic differences in host T cell responses to noninflammatory Ags compared with responses to pathogens. We explain the delayed CD8(+) response by proposing that there is a balance between cross-presentation of Ag by helper cell-licensed dendritic cells, on the one hand, and veto suppression by live male lymphocytes on the other. PMID- 17911585 TI - Discriminating between different pathways of memory CD8+ T cell differentiation. AB - Despite the rapid accumulation of quantitative data on the dynamics of CD8(+) T cell responses following acute viral or bacterial infections of mice, the pathways of differentiation of naive CD8(+) T cells into memory during an immune response remain controversial. Currently, three models have been proposed. In the "stem cell-associated differentiation" model, following activation, naive T cells differentiate into stem cell-like memory cells, which then convert into terminally differentiated short-lived effector cells. In the "linear differentiation" model, following activation, naive T cells first differentiate into effectors, and after Ag clearance, effectors convert into memory cells. Finally, in the "progressive differentiation" model, naive T cells differentiate into memory or effector cells depending on the amount of specific stimulation received, with weaker stimulation resulting in formation of memory cells. This study investigates whether the mathematical models formulated from these hypotheses are consistent with the data on the dynamics of the CD8(+) T cell response to lymphocytic choriomeningitis virus during acute infection of mice. Findings indicate that two models, the stem cell-associated differentiation model and the progressive differentiation model, in which differentiation of cells is strongly linked to the number of cell divisions, fail to describe the data at biologically reasonable parameter values. This work suggests additional experimental tests that may allow for further discrimination between different models of CD8(+) T cell differentiation in acute infections. PMID- 17911586 TI - Differential requirements for OX40 signals on generation of effector and central memory CD4+ T cells. AB - Memory T cells can be divided into effector memory (T(EM)) and central memory (T(CM)) subsets based on their effector function and homing characteristics. Although previous studies have demonstrated that TCR and cytokine signals mediate the generation of the two memory subsets of CD8(+) T cells, the mechanisms for generation of the CD4(+) T(EM) and T(CM) cell subsets are unknown. We found that OX40-deficient mice showed a marked reduction in the number of CD4(+) T(EM) cells, whereas the number of CD4(+) T(CM) cells was normal. Adoptive transfer experiments using Ag-specific CD4(+) T cells revealed that OX40 signals during the priming phase were indispensable for the optimal generation of the CD4(+) T(EM), but not the CD4(+) T(CM) population. In a different transfer experiment with in vitro established CD4(+)CD44(high)CD62L(low) (T(EM) precursor) and CD4(+)CD44(high)CD62L(high) (T(CM) precursor) subpopulations, OX40-KO T(EM) precursor cells could not survive in the recipient mice, whereas wild-type T(EM) precursor cells differentiated into both T(EM) and T(CM) cells. In contrast, T(CM) precursor cells mainly produced T(CM) cells regardless of OX40 signals, implying the dispensability of OX40 for generation of T(CM) cells. Nevertheless, survival of OX40-KO T(EM) cells was partially rescued in lymphopenic mice. During in vitro recall responses, the OX40-KO T(EM) cells that were generated in lymphopenic recipient mice showed impaired cytokine production, suggesting an essential role for OX40 not only on generation but also on effector function of CD4(+) T(EM) cells. Collectively, the present results indicate differential requirements for OX40 signals on generation of CD4(+) T(EM) and T(CM) cells. PMID- 17911587 TI - Antigen presentation by exosomes released from peptide-pulsed dendritic cells is not suppressed by the presence of active CTL. AB - Despite the potency of dendritic cells (DCs) as a vaccine carrier, they are short lived and sensitive to CTL-mediated elimination. Thus, it is believed that the longevity of Ag presentation by peptide-pulsed DC is limited in vivo. Surprisingly, however, we found that although the majority of injected DCs disappeared from the draining lymph nodes within 7 days, Ag presentation persisted for at least 14 days following DC immunization. This prolonged Ag presentation was not mediated by the remaining injected DCs or through Ag transfer to endogenous APCs. We provide evidence that exosomes released by DCs might be responsible for the persistence of Ag presentation. Functional exosomes could be recovered from the draining lymph nodes of C57BL/6 mice following DC vaccination and, in contrast to DCs, T cell stimulation by exosomes in vivo was not affected by the presence of CTL. Our findings demonstrate that Ag presentation following delivery of DC vaccines persists for longer than expected and indicate that the exosome may play a previously unrecognized role in Ag presentation following DC vaccination. Furthermore, our study reinforces the application of exosomes as a vaccination platform and suggests that exosome-based vaccines may be advantageous for booster immunizations due to their resistance to CTL. PMID- 17911588 TI - CD8+ CTL priming by exact peptide epitopes in incomplete Freund's adjuvant induces a vanishing CTL response, whereas long peptides induce sustained CTL reactivity. AB - Therapeutic vaccination trials, in which patients with cancer were vaccinated with minimal CTL peptide in oil-in-water formulations, have met with limited success. Many of these studies were based on the promising data of mice studies, showing that vaccination with a short synthetic peptide in IFA results in protective CD8(+) T cell immunity. By use of the highly immunogenic OVA CTL peptide in IFA as a model peptide-based vaccine, we investigated why minimal CTL peptide vaccines in IFA performed so inadequately to allow full optimization of peptide vaccination. Injection of the minimal MHC class I-binding OVA(257-264) peptide in IFA transiently activated CD8(+) effector T cells, which eventually failed to undergo secondary expansion or to kill target cells, as a result of a sustained and systemic presentation of the CTL peptides gradually leaking out of the IFA depot without systemic danger signals. Complementation of this vaccine with the MHC class II-binding Th peptide (OVA(323-339)) restored both secondary expansion and in vivo effector functions of CD8(+) T cells. Simply extending the CTL peptide to a length of 30 aa also preserved these CD8(+) T cell functions, independent of T cell help, because the longer CTL peptide was predominantly presented in the locally inflamed draining lymph node. Importantly, these functional differences were reproduced in two additional model Ag systems. Our data clearly show why priming of CTL with minimal peptide epitopes in IFA is suboptimal, and demonstrate that the use of longer versions of these CTL peptide epitopes ensures the induction of sustained effector CD8(+) T cell reactivity in vivo. PMID- 17911589 TI - The countervailing actions of myeloid and plasmacytoid dendritic cells control autoimmune diabetes in the nonobese diabetic mouse. AB - Islet Ag-specific CD4(+) T cells receive antigenic stimulation from MHC class II expressing APCs. Herein, we delineate the direct in vivo necessity for distinct subsets of macrophages and dendritic cells (DC) in type 1 diabetes mellitus of the NOD mouse by using diphtheria toxin-mediated cell ablation. The ablation of macrophages had no impact on islet Ag presentation or on the induction of insulitis or diabetes in either transfer or spontaneous models. However, the ablation of CD11b(+)CD11c(+) DC led to the loss of T cell activation, insulitis, and diabetes mediated by CD4(+) T cells. When the specific myeloid DC subset was "added-back" to mice lacking total DC, insulitis and diabetes were restored. Interestingly, when NOD mice were allowed to progress to the insulitis phase, the ablation of DC led to accelerated insulitis. This accelerated insulitis was mediated by the loss of plasmacytoid DC (pDC). When pDC were returned to depleted mice, the localized regulation of insulitis was restored. The loss of pDC in the pancreas itself was accompanied by the localized loss of IDO and the acceleration of insulitis. Thus, CD11c(+)CD11b(+) DC and pDC have countervailing actions in NOD diabetes, with myeloid DC providing critical antigenic stimulation to naive CD4(+) T cells and pDC providing regulatory control of CD4(+) T cell function in the target tissue. PMID- 17911590 TI - The alpha1beta1 integrin and TNF receptor II protect airway CD8+ effector T cells from apoptosis during influenza infection. AB - Primary viral infections of the lung induce potent effector CD8 T cell responses. To function in the influenza-infected airways, CD8 T cells must be able to resist cell death. The majority of the CD8 T cells in the airways and lung parenchyma expressed CD49a, the alpha-chain of the type IV collagen receptor VLA-1, and these cells were highly activated, producing both IFN-gamma and TNF-alpha. In the airways, where type IV collagen is abundant, but not the spleen, the CD49a(+) CD8 cells had reduced proportions of annexin V and caspase 8, and >80% expressed the TNF-alpha receptor II, while Fas, TNFR-I, and CD27 expression were similar to CD49a(-) cells. Furthermore, the CD49a(+), but not CD49a(-), CD8 T cells from the airways were resistant to active induction of apoptosis in the presence of type IV collagen and TNF-alpha in vitro. We propose that TNFR-II and the VLA-1 synergize to protect effector CD8 T cells in the infected airways from apoptosis during the acute infection. PMID- 17911591 TI - PD-1 regulates self-reactive CD8+ T cell responses to antigen in lymph nodes and tissues. AB - PD-1, an inhibitory receptor expressed on activated lymphocytes, regulates tolerance and autoimmunity. We tested the role of PD-1:PD-1 ligand (PD-L) interactions in cross-presentation and the generation and control of CD8(+) responses against self-Ag. Ag-naive PD-1(-/-) OVA-specific OT-I CD8(+) T cells exhibited exacerbated responses to cross-presented Ag in mice expressing soluble OVA under the control of the rat insulin promoter (RIP-ova(high)). Following adoptive transfer into RIP-ova(high) recipients, PD-1(-/-) OT-I T cells expanded in the pancreatic lymph node. In contrast to wild-type OT-I cells, PD-1(-/-) OT-I T cells secreted IFN-gamma and migrated into the pancreas, ultimately causing diabetes. Loss of PD-1 affected CD8(+) cells intrinsically, and did not significantly alter the responses of wild-type OT-I T cells adoptively transferred into the same RIP-ova(high) recipient mouse. PD-1:PD-L interactions also limited CD8(+) effector cells, and PD-L1 expression on parenchymal tissues protected against effector OT-I T cell attack. Finally, we found that the loss of PD-1 on effector OT-I cells lowers the threshold for Ag recognition in peripheral tissues. These findings indicate two checkpoints where PD-1 attenuates self reactive T cell responses: presentation of self-Ag to naive self-reactive T cells by dendritic cells in the draining lymph node and reactivation of pathogenic self reactive T cells in the target organ. PMID- 17911592 TI - The importance of myeloid-derived suppressor cells in the regulation of autoimmune effector cells by a chronic contact eczema. AB - Induction of a chronic eczema is a most efficient therapy for alopecia areata (AA). We had noted a reduction in regulatory T cells during AA induction and wondered whether regulatory T cells may become recruited or expanded during repeated skin sensitization or whether additional regulatory cells account for hair regrowth. AA could not be cured by the transfer of CD4(+)CD25(high) lymph node cells from mice repeatedly treated with a contact sensitizer. This obviously is a consequence of a dominance of freshly activated cells as compared with regulatory CD4(+)CD25(+) T cells. Instead, a population of Gr-1(+)CD11b(+) cells was significantly increased in skin and spleen of AA mice repeatedly treated with a contact sensitizer. Gr-1(+)CD11b(+) spleen cells mostly expressed CD31. Expression of several proinflammatory cytokines as well as of the IFN-gamma receptor and the TNF receptor I were increased. Particularly in the skin, Gr-1(+) cells expressed several chemokines and CCR8 at high levels. Gr-1(+)CD11b(+) cells most potently suppressed AA effector cell proliferation in vitro and promoted partial hair regrowth in vivo. When cocultured with CD4(+) or CD8(+) cells from AA mice, the Gr-1(+)CD11b(+) cells secreted high levels of NO. However, possibly due to high level Bcl-2 protein expression in AA T cells, apoptosis induction remained unaltered. Instead, zeta-chain expression was strongly down-regulated, which was accompanied by a decrease in ZAP70 and ERK1/2 phosphorylation. Thus, a chronic eczema supports the expansion and activation of myeloid suppressor cells that, via zeta-chain down-regulation, contribute to autoreactive T cell silencing in vitro and in vivo. PMID- 17911593 TI - Modulation of miR-155 and miR-125b levels following lipopolysaccharide/TNF-alpha stimulation and their possible roles in regulating the response to endotoxin shock. AB - We report here that miR-155 and miR-125b play a role in innate immune response. LPS stimulation of mouse Raw 264.7 macrophages resulted in the up-regulation of miR-155 and down-regulation of miR-125b levels. The same changes also occurred when C57BL/6 mice were i.p. injected with LPS. Furthermore, the levels of miR-155 and miR-125b in Raw 264.7 cells displayed oscillatory changes in response to TNF alpha. These changes were impaired by pretreating the cells with the proteasome inhibitor MG-132, suggesting that these two microRNAs (miRNAs) may be at least transiently under the direct control of NF-kappaB transcriptional activity. We show that miR-155 most probably directly targets transcript coding for several proteins involved in LPS signaling such as the Fas-associated death domain protein (FADD), IkappaB kinase epsilon (IKKepsilon), and the receptor (TNFR superfamily)-interacting serine-threonine kinase 1 (Ripk1) while enhancing TNF alpha translation. In contrast, miR-125b targets the 3'-untranslated region of TNF-alpha transcripts; therefore, its down-regulation in response to LPS may be required for proper TNF-alpha production. Finally, Emu-miR-155 transgenic mice produced higher levels of TNF-alpha when exposed to LPS and were hypersensitive to LPS/d-galactosamine-induced septic shock. Altogether, our data suggest that the LPS/TNF-alpha-dependent regulation of miR-155 and miR-125b may be implicated in the response to endotoxin shock, thus offering new targets for drug design. PMID- 17911594 TI - CD8 T cell responses to myelin oligodendrocyte glycoprotein-derived peptides in humanized HLA-A*0201-transgenic mice. AB - Multiple sclerosis (MS) is a demyelinating inflammatory disease of the CNS. Though originally believed to be CD4-mediated, additional immune effector mechanisms, including myelin-specific CD8(+) T cells, are now proposed to participate in the pathophysiology of MS. To study the immunologic and encephalitogenic behavior of HLA-A*0201-binding myelin-derived epitopes in vivo, we used a humanized HLA-A*0201-transgenic mouse model. Eight HLA-A*0201-binding peptides derived from myelin oligodendrocyte glycoprotein (MOG), an immunodominant myelin self-Ag, were identified in silico. After establishing their relative affinity for HLA-A*0201 and their capacity to form stable complexes with HLA-A*0201 in vitro, their immunological characteristics were studied in HLA-A*0201-transgenic mice. Five MOG peptides, which bound stably to HLA-A*0201 exhibited strong immunogenicity by inducing a sizeable MOG-specific HLA-A*0201-restricted CD8(+) T cell response in vivo. Of these five candidate epitopes, four were processed by MOG-transfected RMA target cells and two peptides proved immunodominant in vivo in response to a plasmid-encoding native full-length MOG. One of the immunodominant MOG peptides (MOG(181)) generated a cytotoxic CD8(+) T cell response able to aggravate CD4(+)-mediated EAE. Therefore, this detailed in vivo characterization provides a hierarchy of candidate epitopes for MOG-specific CD8(+) T cell responses in HLA-A*0201 MS patients identifying the encephalitogenic MOG(181) epitope as a primary candidate. PMID- 17911595 TI - Role of CXCR5 and CCR7 in follicular Th cell positioning and appearance of a programmed cell death gene-1high germinal center-associated subpopulation. AB - Th cell access to primary B cell follicles is dependent on CXCR5. However, whether CXCR5 induction on T cells is sufficient in determining their follicular positioning has been unclear. In this study, we find that transgenic CXCR5 overexpression is not sufficient to promote follicular entry of naive T cells unless the counterbalancing influence of CCR7 ligands is removed. In contrast, the positioning of Ag-engaged T cells at the B/T boundary could occur in the absence of CXCR5. The germinal center (GC) response was 2-fold reduced when T cells lacked CXCR5, although these T cells were able to access the GC. Finally, CXCR5(high)CCR7(low) T cells were found to have elevated IL-4 transcript and programmed cell death gene-1 (PD-1) expression, and PD-1(high) cells were reduced in the absence of T cell CXCR5 or in mice compromised in GC formation. Overall, these findings provide further understanding of how the changes in CXCR5 and CCR7 expression regulate Th cell positioning during Ab responses, and they suggest that development and/or maintenance of a PD-1(high) follicular Th cell subset is dependent on appropriate interaction with GC B cells. PMID- 17911596 TI - Expression of CD80/86 on B cells is essential for autoreactive T cell activation and the development of arthritis. AB - Depletion of B cells in rheumatoid arthritis is therapeutically efficacious. Yet, the mechanism by which B cells participate in the inflammatory process is unclear. We previously demonstrated that Ag-specific B cells have two important functions in the development of arthritis in a murine model of rheumatoid arthritis, proteoglycan (PG)-induced arthritis (PGIA). PG-specific B cells function as autoantibody-producing cells and as APCs that activate PG-specific T cells. Moreover, the costimulatory molecule CD86 is up-regulated on PG-specific B cells in response to stimulation with PG. To address the requirement for CD80/CD86 expression on B cells in the development of PGIA, we generated mixed bone marrow chimeras in which CD80/CD86 is specifically deleted on B cells and not on other APC populations. Chimeras with a specific deficiency in CD80/CD86 expression on B cells are resistant to the induction of PGIA. The concentration of PG-specific autoantibody is similar in mice sufficient or deficient for CD80/86-expressing B cells, which indicates that resistance to PGIA is not due to the suppression of PG-specific autoantibody production. CD80/86-deficient B cells failed to effectively activate PG-specific autoreactive T cells as indicated by the failure of T cells from PG-immunized CD80/86-deficient B cell chimeras to transfer arthritis into SCID mice. In vitro secondary recall responses to PG are also dependent on CD80/86-expressing B cells. These results demonstrate that a CD80/86:CD28 costimulatory interaction between B cells and T cells is required for autoreactive T cell activation and the induction of arthritis but not for B cell autoantibody production. PMID- 17911597 TI - Unique features and distribution of the chicken CD83+ cell. AB - The central importance of dendritic cells (DC) in both innate and acquired immunity is well recognized in the mammalian immune system. By contrast DC have yet to be characterized in avian species despite the fact that avian species such as the chicken have a well-developed immune system. CD83 has proven to be an excellent marker for DC in human and murine immune systems. In this study we identify chicken CD83 (chCD83) as the avian equivalent of the human and murine DC marker CD83. We demonstrate for the first time that unlike human and murine CD83, chCD83 is uniquely expressed in the B cell areas of secondary lymphoid organs and in organs with no human or murine equivalent such as the bursa and Harderian gland. Furthermore through multicolor immunofluorescence, we identify chCD83(+) populations that have unique attributes akin to both DC and follicular DC. These attributes include colocalization with B cell microenvironments, MHC class II expression, dendritic morphology, and distribution throughout peripheral and lymphoid tissues. PMID- 17911598 TI - Cross-regulation between type I and type II NKT cells in regulating tumor immunity: a new immunoregulatory axis. AB - Negative immunoregulation is a major barrier to successful cancer immunotherapy. The NKT cell is known to be one such regulator. In this study we explored the roles of and interaction between the classical type I NKT cell and the poorly understood type II NKT cell in the regulation of tumor immunity. Selective stimulation of type II NKT cells suppressed immunosurveillance, whereas stimulation of type I NKT cells protected against tumor growth even when responses were relatively skewed toward Th2 cytokines. When both were stimulated simultaneously, type II NKT cells appeared to suppress the activation in vitro and protective effect in vivo of type I NKT cells. In the absence of type I, suppression by type II NKT cells increased, suggesting that type I cells reduce the suppressive effect of type II NKT cells. Thus, in tumor immunity type I and type II NKT cells have opposite and counteractive roles and define a new immunoregulatory axis. Alteration of the balance between the protective type I and the suppressive type II NKT cell may be exploited for therapeutic intervention in cancer. PMID- 17911599 TI - Mycobacterium bovis bacillus Calmette-Guerin secreting active cathepsin S stimulates expression of mature MHC class II molecules and antigen presentation in human macrophages. AB - A successful Th cell response to bacterial infections is induced by mature MHC class II molecules presenting specific Ag peptides on the surface of macrophages. In recent studies, we demonstrated that infection with the conventional vaccine Mycobacterium bovis bacillus Calmette-Guerin (BCG) specifically blocks the surface export of mature class II molecules in human macrophages by a mechanism dependent on inhibition of cathepsin S (Cat S) expression. The present study examined class II expression in macrophages infected with a rBCG strain engineered to express and secrete biologically active human Cat S (rBCG-hcs). Cat S activity was completely restored in cells ingesting rBCG-hcs, which secreted substantial levels of Cat S intracellularly. Thus, infection with rBCG-hcs, but not parental BCG, restored surface expression of mature MHC class II molecules in response to IFN-gamma, presumably as result of MHC class II invariant chain degradation dependent on active Cat S secreted by the bacterium. These events correlated with increased class II-directed presentation of mycobacterial Ag85B to a specific CD4(+) T cell hybridoma by rBCG-hcs-infected macrophages. Consistent with these findings, rBCG-hcs was found to accelerate the fusion of its phagosome with lysosomes, a process that optimizes Ag processing in infected macrophages. These data demonstrated that intracellular restoration of Cat S activity improves the capacity of BCG-infected macrophages to stimulate CD4(+) Th cells. Given that Th cells play a major role in protection against tuberculosis, rBCG-hcs would be a valuable tuberculosis vaccine candidate. PMID- 17911601 TI - Inhibition of T cell activation by cyclic adenosine 5'-monophosphate requires lipid raft targeting of protein kinase A type I by the A-kinase anchoring protein ezrin. AB - cAMP negatively regulates T cell immune responses by activation of type I protein kinase A (PKA), which in turn phosphorylates and activates C-terminal Src kinase (Csk) in T cell lipid rafts. Using yeast two-hybrid screening, far-Western blot, immunoprecipitation and immunofluorescense analyses, and small interfering RNA mediated knockdown, we identified Ezrin as the A-kinase anchoring protein that targets PKA type I to lipid rafts. Furthermore, Ezrin brings PKA in proximity to its downstream substrate Csk in lipid rafts by forming a multiprotein complex consisting of PKA/Ezrin/Ezrin-binding protein 50, Csk, and Csk-binding protein/phosphoprotein associated with glycosphingolipid-enriched microdomains. The complex is initially present in immunological synapses when T cells contact APCs and subsequently exits to the distal pole. Introduction of an anchoring disruptor peptide (Ht31) into T cells competes with Ezrin binding to PKA and thereby releases the cAMP/PKA type I-mediated inhibition of T cell proliferation. Finally, small interfering RNA-mediated knockdown of Ezrin abrogates cAMP regulation of IL-2. We propose that Ezrin is essential in the assembly of the cAMP-mediated regulatory pathway that modulates T cell immune responses. PMID- 17911602 TI - Regulation of Ca2+ signaling in mast cells by tyrosine-phosphorylated and unphosphorylated non-T cell activation linker. AB - Engagement of the FcepsilonRI in mast cells and basophils leads to a rapid tyrosine phosphorylation of the transmembrane adaptors LAT (linker for activation of T cells) and NTAL (non-T cell activation linker, also called LAB or LAT2). NTAL regulates activation of mast cells by a mechanism, which is incompletely understood. Here we report properties of rat basophilic leukemia cells with enhanced or reduced NTAL expression. Overexpression of NTAL led to changes in cell morphology, enhanced formation of actin filaments and inhibition of the FcepsilonRI-induced tyrosine phosphorylation of the FcepsilonRI subunits, Syk kinase and LAT and all downstream activation events, including calcium and secretory responses. In contrast, reduced expression of NTAL had little effect on early FcepsilonRI-induced signaling events but inhibited calcium mobilization and secretory response. Calcium response was also repressed in Ag-activated cells defective in Grb2, a major target of phosphorylated NTAL. Unexpectedly, in cells stimulated with thapsigargin, an inhibitor of the endoplasmic reticulum Ca(2+) ATPase, the amount of cellular NTAL directly correlated with the uptake of extracellular calcium even though no enhanced tyrosine phosphorylation of NTAL was observed. The combined data indicate that NTAL regulates FcepsilonRI-mediated signaling at multiple steps and by different mechanisms. At early stages NTAL interferes with tyrosine phosphorylation of several substrates and formation of signaling assemblies, whereas at later stages it regulates the activity of store operated calcium channels through a distinct mechanism independent of enhanced NTAL tyrosine phosphorylation. PMID- 17911600 TI - Hydrodynamic vaccination with DNA encoding an immunologically privileged retinal antigen protects from autoimmunity through induction of regulatory T cells. AB - The eye is an immunologically privileged organ whose Ags serve as targets for experimental autoimmune uveitis (EAU), a model for human uveitis. We used a hydrodynamic i.v. injection of naked DNA to express the uveitogenic retinal Ag interphotoreceptor retinoid-binding protein (IRBP) in the periphery, thus revoking its immune-privileged status. IRBP was expressed in the liver within hours of administration of as little as 10 microg of IRBP-DNA. Vaccinated mice were highly protected from EAU induced by immunization with IRBP for at least 10 wk after vaccination. Protection was partial in a reversal protocol. Mechanistic studies revealed specific hyporesponsiveness to IRBP without immune deviation, no evidence for apoptosis either by the Fas- or Bcl-2-regulated (mitochondrial) pathway and apparent lack of dependence on CD8(+) cells, IL-10, or TGF-beta. In contrast, depletion of CD25(+) cells after vaccination and before challenge markedly abrogated protection. IRBP-specific CD4(+)CD25(high) T cells could be cultured from vaccinated mice and transferred protection to unvaccinated, EAU challenged recipients. In vitro characterization of these cells revealed that they are Ag specific, anergic, express FoxP3, CTLA-4, and glucocorticoid-induced TNFR, and suppress by contact. Thus, expression of IRBP in the periphery by DNA vaccination results in tolerance that acts at least in part through induction of IRBP-specific, FoxP3(+)CD4(+)CD25(+) regulatory T cells. DNA vaccination may offer a new approach to Ag-specific therapy of uveitis. PMID- 17911604 TI - Enhanced engagement of CTLA-4 induces antigen-specific CD4+CD25+Foxp3+ and CD4+CD25- TGF-beta 1+ adaptive regulatory T cells. AB - CTLA-4 is a critical negative regulator of T cell response and is instrumental in maintaining immunological tolerance. In this article, we report that enhanced selective engagement of CTLA-4 on T cells by Ag-presenting dendritic cells resulted in the induction of Ag-specific CD4(+)CD25(+)Foxp3(+) and CD4(+)CD25( )TGF-beta1(+) adaptive Tregs. These cells were CD62L(low) and hyporesponsive to stimulation with cognate Ag but demonstrated a superior ability to suppress Ag specific effector T cell response compared with their CD62L(high) counterparts. Importantly, treatment of mice with autoimmune thyroiditis using mouse thyroglobulin (mTg)-pulsed anti-CTLA-4 agonistic Ab-coated DCs, which results in a dominant engagement of CTLA-4 upon self-Ag presentation, not only suppressed thyroiditis but also prevented reemergence of the disease upon rechallenge with mTg. Further, the disease suppression was associated with significantly reduced mTg-specific T cell and Ab responses. Collectively, our results showed an important role for selective CTLA-4 signaling in the induction of adaptive Tregs and suggested that approaches that allow dominant CTLA-4 engagement concomitant with Ag-specific TCR ligation can be used for targeted therapy. PMID- 17911603 TI - Silent development of memory progenitor B cells. AB - T cell-dependent immune responses generate long-lived plasma cells and memory B cells, both of which express hypermutated Ab genes. The relationship between these cell types is not entirely understood. Both appear to emanate from the germinal center reaction, but it is unclear whether memory cells evolve while obligatorily generating plasma cells by siblings under all circumstances. In the experiments we report, plasma cell development was functionally segregated from memory cell development by a series of closely spaced injections of Ag delivered during the period of germinal center development. The injection series elevated serum Ab of low affinity, supporting the idea that a strong Ag signal drives plasma cell development. At the same time, the injection series produced a distinct population of affinity/specificity matured memory B cells that were functionally silent, as manifested by an absence of corresponding serum Ab. These cells could be driven by a final booster injection to develop into Ab-forming cells. This recall response required that a rest period precede the final booster injection, but a pause of only 4 days was sufficient. Our results support a model of memory B cell development in which extensive affinity/specificity maturation can take place within a B cell clone under some circumstances in which a concomitant generation of Ab-forming cells by siblings does not take place. PMID- 17911606 TI - A link between PDL1 and T regulatory cells in fetomaternal tolerance. AB - Acceptance of the fetus expressing allogeneic paternal Ags by the mother is a physiologic model of transplantation tolerance. Various mechanisms contribute to fetal evasion from immune attack by maternal leukocytes. We have recently demonstrated that the inhibitory costimulatory molecule PDL1 plays a critical role in fetomaternal tolerance in that PDL1 blockade or deficiency resulted in decreased allogeneic fetal survival rates. CD4(+)CD25(+) T regulatory cells (Tregs) have also been demonstrated to play an important role in fetomaternal tolerance. Since PDL1 is expressed on Tregs, we explored the interactions between PDL1 and Tregs in vivo in a mouse model of fetomaternal tolerance. Depletion of CD25(+) T cells abrogated the effect of anti-PDL1 Ab indicating that the effect of PDL1 is possibly mediated by CD25(+) Tregs. Adoptive transfer of Tregs from wild-type but not PDL1-deficient mice into PDL1-deficient recipients significantly improved fetal survival. The frequency, phenotype and placental trafficking of Tregs from PDL1-deficient mice were similar to those of wild-type controls, but were defective in inhibiting alloreactive Th1 cells in vitro. This is the first report providing evidence for a link between PDL1 and T regulatory cells in mediating fetomaternal tolerance. PMID- 17911605 TI - PDL1 is required for peripheral transplantation tolerance and protection from chronic allograft rejection. AB - The PD-1:PDL pathway plays an important role in regulating alloimmune responses but its role in transplantation tolerance is unknown. We investigated the role of PD-1:PDL costimulatory pathway in peripheral and a well established model of central transplantation tolerance. Early as well as delayed blockade of PDL1 but not PDL2 abrogated tolerance induced by CTLA4Ig in a fully MHC-mismatched cardiac allograft model. Accelerated rejection was associated with a significant increase in the frequency of IFN-gamma-producing alloreactive T cells and expansion of effector CD8(+) T cells in the periphery, and a decline in the percentage of Foxp3(+) graft infiltrating cells. Similarly, studies using PDL1/L2-deficient recipients confirmed the results with Ab blockade. Interestingly, while PDL1 deficient donor allografts were accepted by wild-type recipients treated with CTLA4Ig, the grafts developed severe chronic rejection and vasculopathy when compared with wild-type grafts. Finally, in a model of central tolerance induced by mixed allogeneic chimerism, engraftment was not abrogated by PDL1/L2 blockade. These novel data demonstrate the critical role of PDL1 for induction and maintenance of peripheral transplantation tolerance by its ability to alter the balance between pathogenic and regulatory T cells. Expression of PDL1 in donor tissue is critical for prevention of in situ graft pathology and chronic rejection. PMID- 17911607 TI - High susceptibility of human dendritic cells to avian influenza H5N1 virus infection and protection by IFN-alpha and TLR ligands. AB - There is worldwide concern that the avian influenza H5N1 virus, with a mortality rate of >50%, might cause the next influenza pandemic. Unlike most other influenza infections, H5N1 infection causes a systemic disease. The underlying mechanisms for this effect are still unclear. In this study, we investigate the interplay between avian influenza H5N1 and human dendritic cells (DC). We showed that H5N1 virus can infect and replicate in monocyte-derived and blood myeloid DC, leading to cell death. These results suggest that H5N1 escapes viral-specific immunity, and could disseminate via DC. In contrast, blood pDC were resistant to infection and produced high amounts of IFN-alpha. Addition of this cytokine to monocyte-derived DC or pretreatment with TLR ligands protected against infection and the cytopathic effects of H5N1 virus. PMID- 17911608 TI - CD11b+Ly-6C(hi) suppressive monocytes in experimental autoimmune encephalomyelitis. AB - Innate immune cells may regulate adaptive immunity by balancing different lineages of T cells and providing negative costimulation. In addition, CD11b(+)Gr 1(+) myeloid-derived suppressor cells have been described in tumor, parasite infection, and severe trauma models. In this study, we observe that splenic CD11b(+) cells markedly increase after experimental autoimmune encephalomyelitis (EAE) immunization, and they suppress T cell proliferation in vitro. Although >80% of CD11b(+) cells express varying levels of Gr-1, only a small population of CD11b(+)Ly-6C(high) inflammatory monocytes (IMC) can efficiently suppress T cell proliferation and induce T cell apoptosis through the production of NO. IFN-gamma produced by activated T cells is essential to induce IMC suppressive function. EAE immunization increases the frequencies of IMC in the bone marrow, spleen, and blood, but not in the lymph nodes. At the peak of EAE, IMC represent approximately 30% of inflammatory cells in the CNS. IMC express F4/80 and CD93 but not CD31, suggesting that they are immature monocytes. Furthermore, IMC have the plasticity to up-regulate NO synthase 2 or arginase 1 expression upon different cytokine treatments. These findings indicate that CD11b(+)Ly-6C(high) IMC induced during EAE priming are powerful suppressors of activated T cells. Further understanding of suppressive monocytes in autoimmune disease models may have important clinical implications for human autoimmune diseases. PMID- 17911609 TI - Antibody-mediated rejection of cardiac allografts in CCR5-deficient recipients. AB - Rejected MHC-mismatched cardiac allografts in CCR5(-/-) recipients have low T cell infiltration, but intense deposition of C3d in the large vessels and capillaries of the graft, characteristics of Ab-mediated rejection. The roles of donor-specific Ab and CD4 and CD8 T cell responses in the rejection of complete MHC-mismatched heart grafts by CCR5(-/-) recipients were directly investigated. Wild-type C57BL/6 and B6.CCR5(-/-) (H-2(b)) recipients of A/J (H-2(a)) cardiac allografts had equivalent numbers of donor-reactive CD4 T cells producing IFN gamma, whereas CD4 T cells producing IL-4 were increased in CCR5(-/-) recipients. Numbers of donor-reactive CD8 T cells producing IFN-gamma were reduced 60% in CCR5(-/-) recipients. Day 8 posttransplant serum titers of donor-specific Ab were 15- to 25-fold higher in CCR5(-/-) allograft recipients, and transfer of this serum provoked cardiac allograft rejection in RAG-1(-/-) recipients within 14 days, whereas transfer of either serum from wild-type recipients or immune serum from CCR5-deficient recipients diluted to titers observed in wild-type recipients did not mediate this rejection. Wild-type C57BL/6 and B6.CCR5(-/-) recipients rejected A/J cardiac grafts by day 11, whereas rejection was delayed (day 12-60, mean 21 days) in muMT(-/-)/CCR5(-/-) recipients. These results indicate that the donor-specific Ab produced in CCR5(-/-) heart allograft recipients is sufficient to directly mediate graft rejection, and the absence of recipient CCR5 expression has differential effects on the priming of alloreactive CD4 and CD8 T cells. PMID- 17911610 TI - Induction of indoleamine 2,3-dioxygenase in vascular smooth muscle cells by interferon-gamma contributes to medial immunoprivilege. AB - Atherosclerosis and graft arteriosclerosis are characterized by leukocytic infiltration of the vessel wall that spares the media. The mechanism(s) for medial immunoprivilege is unknown. In a chimeric humanized mouse model of allograft rejection, medial immunoprivilege was associated with expression of IDO by vascular smooth muscle cells (VSMCs) of rejecting human coronary artery grafts. Inhibition of IDO by 1-methyl-tryptophan (1-MT) increased medial infiltration by allogeneic T cells and increased VSMC loss. IFN-gamma-induced IDO expression and activity in cultured human VSMCs was considerably greater than in endothelial cells (ECs) or T cells. IFN-gamma-treated VSMCs, but not untreated VSMCs nor ECs with or without IFN-gamma pretreatment, inhibited memory Th cell alloresponses across a semipermeable membrane in vitro. This effect was reversed by 1-MT treatment or tryptophan supplementation and replicated by the absence of tryptophan, but not by addition of tryptophan metabolites. However, IFN-gamma treated VSMCs did not activate allogeneic memory Th cells, even after addition of 1-MT or tryptophan. Our work extends the concept of medial immunoprivilege to include immune regulation, establishes the compartmentalization of immune responses within the vessel wall due to distinct microenvironments, and demonstrates a duality of stimulatory EC signals versus inhibitory VSMC signals to artery-infiltrating T cells that may contribute to the chronicity of arteriosclerotic diseases. PMID- 17911611 TI - All-or-none activation of CRAC channels by agonist elicits graded responses in populations of mast cells. AB - In nonexcitable cells, receptor stimulation evokes Ca(2+) release from the endoplasmic reticulum stores followed by Ca(2+) influx through store-operated Ca(2+) channels in the plasma membrane. In mast cells, store-operated entry is mediated via Ca(2+) release-activated Ca(2+) (CRAC) channels. In this study, we find that stimulation of muscarinic receptors in cultured mast cells results in Ca(2+)-dependent activation of protein kinase Calpha and the mitogen activated protein kinases ERK1/2 and this is required for the subsequent stimulation of the enzymes Ca(2+)-dependent phospholipase A(2) and 5-lipoxygenase, generating the intracellular messenger arachidonic acid and the proinflammatory intercellular messenger leukotriene C(4). In cell population studies, ERK activation, arachidonic acid release, and leukotriene C(4) secretion were all graded with stimulus intensity. However, at a single cell level, Ca(2+) influx was related to agonist concentration in an essentially all-or-none manner. This paradox of all or-none CRAC channel activation in single cells with graded responses in cell populations was resolved by the finding that increasing agonist concentration recruited more mast cells but each cell responded by generating all-or-none Ca(2+) influx. These findings were extended to acutely isolated rat peritoneal mast cells where muscarinic or P2Y receptor stimulation evoked all-or-none activation of Ca(2+)entry but graded responses in cell populations. Our results identify a novel way for grading responses to agonists in immune cells and highlight the importance of CRAC channels as a key pharmacological target to control mast cell activation. PMID- 17911612 TI - Dynamic changes in accessibility, nuclear positioning, recombination, and transcription at the Ig kappa locus. AB - The 3-megabase Igkappa locus undergoes differentially controlled nuclear positioning events and chromatin structural changes during the course of B cell development. The temporal association of chromatin structural changes, transcription, and recombination at the Igkappa locus was determined in a murine pre-B cell line that can be induced to recombine at the Igkappa locus and in ex vivo-cultured murine pre-B cells. Additionally, the timing of nuclear positioning relative to the temporal order of chromatin structural changes and recombination and transcription was determined. We demonstrate that before induction, the Igkappa locus was poised for recombination; both alleles were in a contracted state, and the enrichment of histone modifications and germline transcripts of specific Vkappa genes were observed. Histone modifications of the Vkappa genes did not vary upon induction but the levels of modifications correlated with the levels of germline Vkappa gene transcripts and recombination. Upon induction, but before VkappaJkappa recombination, centromeric recruitment of single Igkappa alleles occurred. DNase I sensitivity of the entire locus increased gradually over the course of differentiation while the enrichment of histone modifications downstream of the Vkappa genes was increased in the silencer regions upstream of Jkappa1, within the Igkappa sterile transcript, the kappa constant region, the Ekappai and Ekappa3' enhancers, and the recombining sequence. The ex vivo pre-B cells showed similar patterns of histone modifications across the locus except at the Vkappa genes. In this study, H3 acetylation correlated with levels of germline transcripts while H3 methylation correlated with levels of recombination. PMID- 17911613 TI - Role of activation-induced deaminase protein kinase A phosphorylation sites in Ig gene conversion and somatic hypermutation. AB - Activation-induced deaminase (AID) is thought to initiate somatic hypermutation (SHM), gene conversion (GCV), and class switch recombination (CSR) by the transcription-coupled deamination of cytosine residues in Ig genes. Phosphorylation of AID by protein kinase A (PKA) and subsequent interaction of AID with replication protein A (RPA) have been proposed to play important roles in allowing AID to deaminate DNA during transcription. Serine 38 (S38) of mouse AID is phosphorylated in vivo and lies in a consensus target site for PKA, and mutation of this residue interferes with CSR and SHM. In this study, we demonstrate that S38 in mouse and chicken AID is phosphorylated in chicken DT40 cells and is required for efficient GCV and SHM in these cells. Paradoxically, zebra fish AID, which lacks a serine at the position corresponding to S38, has previously been shown to be active for CSR and we demonstrate that it is active for GCV/SHM. Aspartate 44 (D44) of zebra fish AID has been proposed to compensate for the absence of the S38 phosphorylation site but we demonstrate that mutation of D44 has no effect on GCV/SHM. Some features of zebra fish AID other than D44 might compensate for the absence of S38. Alternatively, the zebra fish protein might function in a manner that is independent of PKA and RPA in DT40 cells, raising the possibility that, under some circumstances, AID mediates efficient Ig gene diversification without the assistance of RPA. PMID- 17911614 TI - Protein kinase C regulates expression and function of inhibitory killer cell Ig like receptors in NK cells. AB - The inhibitory killer cell Ig-like receptors (KIR) negatively regulate NK cell cytotoxicity by activating the Src homology 2 domain-containing protein tyrosine phosphatases 1 and 2 following ligation with MHC class I molecules expressed on normal cells. This requires tyrosine phosphorylation of KIR on ITIMs in the cytoplasmic domain. Surprisingly, we have found that KIR3DL1 is strongly and constitutively phosphorylated on serine and weakly on threonine residues. In this study, we have mapped constitutive phosphorylation sites for casein kinases, protein kinase C, and an unidentified kinase on the KIR cytoplasmic domain. Three of these phosphorylation sites are highly conserved in human inhibitory KIR. Functional studies of the wild-type receptor and serine/threonine mutants indicated that phosphorylation of Ser(394) by protein kinase C slightly suppresses KIR3DL1 inhibitory function, and reduces receptor internalization and turnover. Our results provide evidence that serine/threonine phosphorylation is an important regulatory mechanism of KIR function. PMID- 17911615 TI - A Fas-associated death domain protein/caspase-8-signaling axis promotes S-phase entry and maintains S6 kinase activity in T cells responding to IL-2. AB - Fas-associated death domain protein (FADD) constitutes an essential component of TNFR-induced apoptotic signaling. Paradoxically, FADD has also been shown to be crucial for lymphocyte development and activation. In this study, we report that FADD is necessary for long-term maintenance of S6 kinase (S6K) activity. S6 phosphorylation at serines 240 and 244 was only observed after long-term stimulation of wild-type cells, roughly corresponding to the time before S-phase entry, and was poorly induced in T cells expressing a dominantly interfering form of FADD (FADDdd), viral FLIP, or possessing a deficiency in caspase-8. Defects in S6K1 phosphorylation were also observed. However, defective S6K1 phosphorylation was not a consequence of a wholesale defect in mammalian target of rapamycin function, because 4E-BP1 phosphorylation following T cell activation was unaffected by FADDdd expression. Although cyclin D3 up-regulation and retinoblastoma hypophosphorylation occurred normally in FADDdd T cells, cyclin E expression and cyclin-dependent kinase 2 activation were markedly impaired in FADDdd T cells. These results demonstrate that a FADD/caspase-8-signaling axis promotes T cell cycle progression and sustained S6K activity. PMID- 17911616 TI - STAT5 is essential for Akt/p70S6 kinase activity during IL-2-induced lymphocyte proliferation. AB - IL-2R activates two distinct signaling pathways mediated by the adaptor protein Shc and the transcription factor STAT5. Prior mutagenesis studies of the IL-2R have indicated that the Shc and STAT5 pathways are redundant in the ability to induce lymphocyte proliferation. Yet paradoxically, T cells from STAT5-deficient mice fail to proliferate in response to IL-2, suggesting that the Shc pathway is unable to promote mitogenesis in the genetic absence of STAT5. Here we show in the murine lymphocyte cell line Ba/F3 that low levels of STAT5 activity are essential for Shc signaling. In the absence of STAT5 activity, Shc was unable to sustain activation of the Akt/p70S6 kinase pathway or promote lymphocyte proliferation and viability. Restoring STAT5 activity via a heterologous receptor rescued Shc-induced Akt/p70S6 kinase activity and cell proliferation with kinetics consistent with a transcriptional mechanism. Thus, STAT5 appears to regulate the expression of one or more unidentified components of the Akt pathway. Our results not only explain the severe proliferative defect in STAT5 deficient T cells but also provide mechanistic insight into the oncogenic properties of STAT5 in various leukemias and lymphomas. PMID- 17911618 TI - A role for EZH2 in silencing of IFN-gamma inducible MHC2TA transcription in uveal melanoma. AB - We investigated the contribution of epigenetic mechanisms in MHC2TA transcriptional silencing in uveal melanoma. Although no correlation was observed between impaired CIITA transcript levels after IFN-gamma induction and DNA methylation of MHC2TA promoter IV (CIITA-PIV), an association was found with high levels of trimethylated histone H3-lysine 27 (3Me-K27-H3) in CIITA-PIV chromatin. The 3Me-K27-H3 modification correlated with a strong reduction in RNA polymerase II-recruitment to CIITA-PIV. Interestingly, we observed that none of these epigenetic modifications affected recruitment of activating transcription factors to this promoter. Subsequently, we demonstrated the presence of the histone methyltransferase EZH2 in CIITA-PIV chromatin, which is known to be a component of the Polycomb repressive complex 2 and able to triple methylate histone H3 lysine 27. RNA interference-mediated down-regulation of EZH2 expression resulted in an increase in CIITA transcript levels after IFN-gamma induction. Our data therefore reveal that EZH2 contributes to silencing of IFN-gamma-inducible transcription of MHC2TA in uveal melanoma cells. PMID- 17911617 TI - Analysis of epitope-specific immune responses induced by vaccination with structurally folded and unfolded recombinant Bet v 1 allergen derivatives in man. AB - Previously, we have constructed recombinant derivatives of the major birch pollen allergen, Bet v 1, with a more than 100-fold reduced ability to induce IgE mediated allergic reactions. These derivatives differed from each other because the two recombinant Bet v 1 fragments represented unfolded molecules whereas the recombinant trimer resembled most of the structural fold of the Bet v 1 allergen. In this study, we analyzed the Ab (IgE, IgG subclass, IgA, IgM) response to Bet v 1, recombinant and synthetic Bet v 1-derived peptides in birch pollen allergic patients who had been vaccinated with the derivatives or adjuvant alone. Furthermore, we studied the induction of IgE-mediated skin responses in these patients using Bet v 1 and Bet v 1 fragments. Both types of vaccines induced a comparable IgG1 and IgG4 response against new sequential epitopes which overlap with the conformational IgE epitopes of Bet v 1. This response was 4- to 5-fold higher than that induced by immunotherapy with birch pollen extract. Trimer more than fragments induced also IgE responses against new epitopes and a transient increase in skin sensitivity to the fragments at the beginning of therapy. However, skin reactions to Bet v 1 tended to decrease one year after treatment in both actively treated groups. We demonstrate that vaccination with folded and unfolded recombinant allergen derivatives induces IgG Abs against new epitopes. These data may be important for the development of therapeutic as well as prophylactic vaccines based on recombinant allergens. PMID- 17911619 TI - A population of CD19highCD45R-/lowCD21low B lymphocytes poised for spontaneous secretion of IgG and IgA antibodies. AB - Ab responses to selected Ags are produced by discrete B cell populations whose presence and functional relevance vary along the ontogeny. The earliest B lineage restricted precursors in gestational day 11 mouse embryos display the CD19(+)CD45R/B220(-) phenotype. Phenotypically identical cells persist throughout gestation and in postnatal life, in parallel to the later-arising, CD19(+)CD45R(+) B cells. Very early after birth, the CD19(+)CD45R(-) B cell subset included high frequencies of spontaneously Ig-secreting cells. In the adult spleen, a small subset of CD19(high)CD45R(-/low)IgM(+/-)IgD(-)CD21/Cr2( /low) cells, which was detected in perifollicular areas, displayed genetic and phenotypical traits of highly differentiated B cells, and was enriched in IgG- and IgA-secreting plasma cells. In vitro differentiation and in vivo adoptive transfer experiments of multipotent hemopoietic progenitors revealed that these CD19(high)CD45R(-/low) B cells were preferentially regenerated by embryo-, but not by adult bone marrow-, derived progenitors, except when the latter were inoculated into newborn mice. Both the early ontogenical emergence and the natural production of serum Igs, are shared features of this CD19(high)CD45R( /low) B cell population with innate-like B lymphocytes such as B1 and marginal zone B cells, and suggest that the new population might be related to that category. PMID- 17911620 TI - Expression profiling of immature thymocytes revealed a novel homeobox gene that regulates double-negative thymocyte development. AB - Intrathymic development of CD4/CD8 double-negative (DN) thymocytes can be tracked by well-defined chronological subsets of thymocytes, and is an ideal target of gene expression profiling analysis to clarify the genetic basis of mature T cell production, by which differentiation of immature thymocytes is investigated in terms of gene expression profiles. In this study, we show that development of murine DN thymocytes is predominantly regulated by largely repressive rather than inductive activities of transcriptions, where lineage-promiscuous gene expression in immature thymocytes is down-regulated during their differentiation. Functional mapping of genes showing common temporal expression profiles implicates previously uncharacterized gene regulations that may be relevant to early thymocytes development. A small minority of genes is transiently expressed in the CD44(low)CD25(+) subset of DN thymocytes, from which we identified a novel homeobox gene, Duxl, whose expression is up-regulated by Runx1. Duxl promotes the transition from CD44(high)CD25(+) to CD44(low)CD25(+) in DN thymocytes, while constitutive expression of Duxl inhibits expression of TCR beta-chains and leads to impaired beta selection and greatly reduced production of CD4/CD8 double positive thymocytes, indicating its critical roles in DN thymocyte development. PMID- 17911621 TI - The acquired immune response to the mucosal adjuvant LTK63 imprints the mouse lung with a protective signature. AB - LTK63, a nontoxic mutant of Escherichia coli heat labile enterotoxin (LT), is a potent and safe mucosal adjuvant that has also been shown to confer generic protection to several respiratory pathogens. To understand the mechanisms of action underlying the LTK63 protective effect, we analyzed the molecular and cellular events triggered by its administration in vivo. We show here that LTK63 intrapulmonary administration induced in the mouse lung a specific gene expression signature characterized by the up-regulation of cell cycle genes, several host defense genes, chemokines, chemokine receptors, and immune cell associated genes. Such a transcriptional profile reflected the activation of alveolar macrophages and the recruitment to the lung of T and B cells and innate immune cells such as granulocytes, NK, and dendritic cells. All of these events were T cell dependent and specific for LTK63 because they were absent in SCID and nude mice. Additionally, we showed that LTK63 induces a potent adaptive immune response against itself directed to the lung. We propose that acquired response to LTK63 is the driving force for the local recruitment of both adaptive and innate immune cells. Our data suggest that LTK63 acts as an airway infection mimic that establishes a generic protective environment limiting respiratory infection by innate immune mechanisms and by improving adaptive responses to invading pathogens. PMID- 17911622 TI - Lymphotoxin alpha beta 2 (membrane lymphotoxin) is critically important for resistance to Leishmania major infection in mice. AB - Although the essential role of TNF-alpha in the control of intracellular pathogens including Leishmania major is well established, it is uncertain whether the related cytokine lymphotoxin alphabeta2 (LTalpha1beta2, membrane lymphotoxin) plays any role in this process. In this study, we investigated the contribution of membrane lymphotoxin in host response to L. major infection by using LTbeta deficient (LTbeta(-/-)) mice on the resistant C57BL/6 background. Despite mounting early immune responses comparable to those of wild-type (WT) mice, LTbeta(-/-) mice developed chronic nonhealing cutaneous lesions due to progressive and unresolving inflammation that is accompanied by uncontrolled parasite proliferation. This chronic disease was associated with striking reduction in IL-12 and Ag-specific IFN-gamma production by splenocytes from infected mice. Consistent with defective cellular immune response, infected LTbeta(-/-) mice had significantly low Ag-specific serum IgG1 and IgG2a levels compared with WT mice. Although administration of rIL-12 to L. major-infected LTbeta(-/-) mice caused complete resolution of chronic lesions, it only partially (but significantly) reduced parasite proliferation. In contrast, blockade of LIGHT signaling in infected LTbeta(-/-) mice resulted in acute and progressive lesion development, massive parasite proliferation, and dissemination to the visceral organs. Although infected LTbeta(-/-) WT bone marrow chimeric mice were more resistant than LTbeta(-/-) mice, they still had reduced ability to control parasites and showed defective IL-12 and IFN-gamma production compared with infected WT mice. These results suggest that membrane lymphotoxin plays critical role in resistance to L. major by promoting effective T cell-mediated anti Leishmania immunity. PMID- 17911623 TI - IL-13 induces disease-promoting type 2 cytokines, alternatively activated macrophages and allergic inflammation during pulmonary infection of mice with Cryptococcus neoformans. AB - In the murine model of Cryptococcus neoformans infection Th1 (IL-12/IFN-gamma) and Th17 (IL-23/IL-17) responses are associated with protection, whereas an IL-4 dependent Th2 response exacerbates disease. To investigate the role of the Th2 cytokine IL-13 during pulmonary infection with C. neoformans, IL-13 overexpressing transgenic (IL-13Tg(+)), IL-13-deficient (IL-13(-/-)), and wild type (WT) mice were infected intranasally. Susceptibility to C. neoformans infection was found when IL-13 was induced in WT mice or overproduced in IL 13Tg(+) mice. Infected IL-13Tg(+) mice had a reduced survival time and higher pulmonary fungal load as compared with WT mice. In contrast, infected IL-13(-/-) mice were resistant and 89% of these mice survived the entire period of the experiment. Ag-specific production of IL-13 by susceptible WT and IL-13Tg(+) mice was associated with a significant type 2 cytokine shift but only minor changes in IFN-gamma production. Consistent with enhanced type 2 cytokine production, high levels of serum IgE and low ratios of serum IgG2a/IgG1 were detected in susceptible WT and IL-13Tg(+) mice. Interestingly, expression of IL-13 by susceptible WT and IL-13Tg(+) mice was associated with reduced IL-17 production. IL-13 was found to induce formation of alternatively activated macrophages expressing arginase-1, macrophage mannose receptor (CD206), and YM1. In addition, IL-13 production led to lung eosinophilia, goblet cell metaplasia and elevated mucus production, and enhanced airway hyperreactivity. This indicates that IL-13 contributes to fatal allergic inflammation during C. neoformans infection. PMID- 17911624 TI - The C/EBP beta isoform 34-kDa LAP is responsible for NF-IL-6-mediated gene induction in activated macrophages, but is not essential for intracellular bacteria killing. AB - The C/ebpb gene is translated into three different protein isoforms, two transcriptional activating proteins (38-kDa Full and 34-kDa liver-enriched transcriptional activation protein (LAP)) and one transcriptional inhibitory protein, by alternative use of different AUG initiation codons within the same open reading frame. The isoform 34-kDa LAP is thought to be the most transcriptionally active form of C/EBPbeta in macrophages. To assess the function of the 34-kDa LAP in vivo, we generated knock-in mice, in which methionine 20 of C/EBPbeta, the start site for the 34-kDa LAP is replaced with an alanine. The expression of the 34-kDa LAP was abolished in C/ebpb(M20A/M20A) mice. The induction of C/EBPbeta target genes, such as inflammatory cytokines, chemokines, prostanoid synthetase, and antimicrobial peptides, was abolished in C/ebpb(M20A/M20A) macrophages, and C/ebpb(M20A/M20A) mice were susceptible to Listeria monocytogenes infection. Furthermore, the heat-killed Propionibacterium acnes-induced Th1 response, granuloma formation, and LPS shock were severely impaired. Nevertheless, impairment of intracellular bacteria killing, which is the most prominent phenotype in C/EBPbeta-deficient mice, was not observed in C/ebpb(M20A/M20A) mice. Collectively, we demonstrated that 34-kDa LAP is responsible for NF-IL6-mediated gene induction, but not essential for intracellular bacteria killing in activated macrophages. PMID- 17911625 TI - Bone marrow of persistently hepatitis C virus-infected individuals accumulates memory CD8+ T cells specific for current and historical viral antigens: a study in patients with benign hematological disorders. AB - The role of virus-specific T cells in hepatitis C virus (HCV) pathogenesis is not clear. Existing knowledge on the frequency, phenotype, and behavior of these cells comes from analyses of blood and liver, but other lymphoid compartments that may be important sites for functionally mature T cells have not yet been analyzed. We studied HCV-specific T cells from bone marrow, in comparison to those from peripheral blood and liver biopsy tissue, from 20 persistently HCV infected patients with benign hematological disorders. Bone marrow contained a sizeable pool of CD8(+) T cells specific for epitopes from structural and nonstructural HCV proteins. These cells displayed the same effector memory phenotype as liver-derived equivalents and the same proliferative potential as blood-derived equivalents but had greater antiviral effector functions such as Ag specific cytotoxicity and IFN-gamma production. These features were not shared by influenza virus-specific CD8(+) T cells in the same bone marrow samples. Despite their highly differentiated phenotype and activated status, some bone marrow resident HCV-specific CD8(+) T cells were not directed against the infecting virus but, instead, against historical HCV Ags (i.e., viral species of a previous infection or minor viral species of the current infection). These findings provide a snapshot view of the distribution, differentiation, and functioning of virus-specific memory T cells in patients with persistent HCV infection. PMID- 17911627 TI - Gr-1high polymorphonuclear leukocytes and NK cells act via IL-15 to clear intracellular Haemophilus influenzae in experimental murine peritonitis and pneumonia. AB - Polymorphonuclear leukocytes (PMNs) can be divided into Gr-1(high) and Gr-1(low) subpopulations, but the differences in the functions of these cells in the host are unknown. This study investigated the roles of these two cell populations in the clearance of an intracellular pathogen (Haemophilus influenzae) causing murine peritonitis and pneumonia. Microarray analysis and quantitative real-time PCR analysis of proteose peptone-elicited peritoneal murine PMNs showed that IL 15 mRNA levels were significantly higher in Gr-1(high) PMNs than in Gr-1(low) PMNs. In addition, IL-15 was produced only by Gr-1-positive PMNs, especially Gr 1(high) PMNs. IL-15 was required for efficient clearance of experimental murine H. influenzae pneumonia, as 4 days postinfection lungs from IL-15 knockout mice contained 50- to 100-fold more bacteria than did wild-type mouse lungs. Gr-1 PMN depleted C57BL/6 mice were more susceptible to H. influenzae pneumonia than were Gr-1 PMN replete C57BL/6 mice or C57BL/6 nude mice, demonstrating that Gr-1 PMNs are important in the clearance of intracellular bacteria. IL-15-activated NK cells killed H. influenzae in PMNs. Flow cytometry confirmed the expression of CD69 on the cell membrane of IL-15-activated NK cells. Our results show that Gr 1(high) PMNs produce more IL-15 than Gr-1(low) PMNs, and that IL-15-activated NK cells protect against early infection by H. influenzae. PMID- 17911626 TI - The proinflammatory cytokine macrophage migration inhibitory factor regulates glucose metabolism during systemic inflammation. AB - Inflammation provokes significant abnormalities in host metabolism that result from the systemic release of cytokines. An early response of the host is hyperglycemia and resistance to the action of insulin, which progresses over time to increased glucose uptake in peripheral tissue. Although the cytokine TNF-alpha has been shown to exert certain catabolic effects, recent studies suggest that the metabolic actions of TNF-alpha occur by the downstream regulation of additional mediators, such as macrophage migration inhibitory factor (MIF). We investigated the glycemic responses of endotoxemic mice genetically deficient in MIF (MIF(-/-)). In contrast to wild-type mice, MIF(-/-) mice exhibit normal blood glucose and lactate responses following the administration of endotoxin, or TNF alpha. MIF(-/-) mice also show markedly increased glucose uptake into white adipose tissue in vivo in the endotoxemic state. Treatment of adipocytes with MIF, or anti-MIF mAb, modulates insulin-mediated glucose transport and insulin receptor signal transduction; these effects include the phosphorylation of insulin receptor substrate-1, its association with the p85 regulatory subunit of PI3K, and the downstream phosphorylation of Akt. Genetic MIF deficiency also promotes adipogenesis, which is in accord with a downstream role for MIF in the action of TNF-alpha. These studies support an important role for MIF in host glucose metabolism during sepsis. PMID- 17911628 TI - CD8 T cells inhibit respiratory syncytial virus (RSV) vaccine-enhanced disease. AB - Vaccination of children with a formalin-inactivated (FI) respiratory syncytial virus (RSV) vaccine led to exacerbated disease including pulmonary eosinophilia following a natural RSV infection. Immunization of BALB/c mice with FI-RSV or a recombinant vaccinia virus (vv) expressing the RSV attachment (G) protein (vvG) results in a pulmonary Th2 response and eosinophilia after RSV challenge that closely mimics the RSV vaccine-enhanced disease observed in humans. The underlying causes of RSV vaccine-enhanced disease remain poorly understood. We demonstrate here that RSV M2-specific CD8 T cells reduce the Th2-mediated pathology induced by vvG-immunization and RSV challenge in an IFN-gamma independent manner. We also demonstrate that FI-RSV immunization does not induce a measurable RSV-specific CD8 T cell response and that priming FI-RSV-immunized mice for a potent memory RSV-specific CD8 T cell response abrogates pulmonary eosinophilia after subsequent RSV challenge. Our results suggest that the failure of the FI-RSV vaccine to induce a CD8 T cell response may have contributed to the development of pulmonary eosinophilia and augmented disease that occurred in vaccinated individuals. PMID- 17911629 TI - Activation of innate immune defense mechanisms by signaling through RIG-I/IPS-1 in intestinal epithelial cells. AB - Intestinal epithelial cells (IECs) are a first line of defense against microbial pathogens that enter the host through the intestinal tract. Moreover, viral pathogens that infect the host via the intestinal epithelium are an important cause of morbidity and mortality. However, the mechanisms by which viral pathogens activate antiviral defense mechanisms in IECs are largely unknown. The synthetic dsRNA analog polyinosinic-polycytidylic acid and infection with live virus were used to probe the molecules that are activated and the mechanisms of signaling in virus-infected human IECs. Polyinosinic-polycytidylic acid activated IFN regulatory factor 3 dimerization and phosphorylation, increased activity of the IFN-stimulated response element, induced a significant increase in IFN-beta mRNA transcripts and IFN-beta secretion, and up-regulated the expression of IFN regulated genes in IECs. Those responses were dependent upon activation of the dsRNA binding protein retinoic acid inducible gene I (RIG-I) and the RIG-I interacting protein IFN promoter stimulator-1, but not on dsRNA-activated protein kinase or TLR3, which also were expressed by IECs. Virus replication and virus induced cell death increased in IECs in which RIG-I was silenced, consistent with the importance of the RIG-I signaling pathway in IEC antiviral innate immune defense mechanisms. PMID- 17911630 TI - Helicobacter pylori vacuolating cytotoxin inhibits activation-induced proliferation of human T and B lymphocyte subsets. AB - Helicobacter pylori are Gram-negative bacteria that persistently colonize the human gastric mucosa despite the recruitment of immune cells. The H. pylori vacuolating cytotoxin (VacA) recently has been shown to inhibit stimulation induced proliferation of primary human CD4(+) T cells. In this study, we investigated effects of VacA on the proliferation of various other types of primary human immune cells. Intoxication of PBMC with VacA inhibited the stimulation-induced proliferation of CD4(+) T cells, CD8(+) T cells, and B cells. VacA also inhibited the proliferation of purified primary human CD4(+) T cells that were stimulated by dendritic cells. VacA inhibited both T cell-induced and PMA/anti-IgM-induced proliferation of purified B cells. Intoxication with VacA did not alter the magnitude of calcium flux that occurred upon stimulation of CD4(+) T cells or B cells, indicating that VacA does not alter early signaling events required for activation and proliferation. VacA reduced the mitochondrial membrane potential of CD4(+) T cells, but did not reduce the mitochondrial membrane potential of B cells. We propose that the immunomodulatory actions of VacA on T and B lymphocytes, the major effectors of the adaptive immune response, may contribute to the ability of H. pylori to establish a persistent infection in the human gastric mucosa. PMID- 17911631 TI - Regulation of tumor cell sensitivity to TRAIL-induced apoptosis by the metastatic suppressor Raf kinase inhibitor protein via Yin Yang 1 inhibition and death receptor 5 up-regulation. AB - Raf-1 kinase inhibitor protein (RKIP) has been implicated in the regulation of cell survival pathways and metastases, and is poorly expressed in tumors. We have reported that the NF-kappaB pathway regulates tumor resistance to apoptosis by the TNF-alpha family via inactivation of the transcription repressor Yin Yang 1 (YY1). We hypothesized that RKIP overexpression may regulate tumor sensitivity to death ligands via inhibition of YY1 and up-regulation of death receptors (DRs). The TRAIL-resistant prostate carcinoma PC-3 and melanoma M202 cell lines were examined. Transfection with CMV-RKIP, but not with control CMV-EV, sensitized the cells to TRAIL-mediated apoptosis. Treatment with RKIP small interfering RNA (siRNA) inhibited TRAIL-induced apoptosis. RKIP overexpression was paralleled with up-regulation of DR5 transcription and expression; no change in DR4, decoy receptor 1, and decoy receptor 2 expression; and inhibition of YY1 transcription and expression. Inhibition of YY1 by YY1 siRNA sensitized the cells to TRAIL apoptosis concomitantly with DR5 up-regulation. RKIP overexpression inhibited several antiapoptotic gene products such as X-linked inhibitor of apoptosis (XIAP), c-FLIP long, and Bcl-x(L) that were accompanied with mitochondrial membrane depolarization. RKIP overexpression in combination with TRAIL resulted in the potentiation of these above effects and activation of caspases 8, 9, and 3, resulting in apoptosis. These findings demonstrate that RKIP overexpression regulates tumor cell sensitivity to TRAIL via inhibition of YY1, up-regulation of DR5, and modulation of apoptotic pathways. We suggest that RKIP may serve as an immune surveillance cancer gene, and its low expression or absence in tumors allows the tumor to escape host immune cytotoxic effector cells. PMID- 17911632 TI - Specific leukotriene receptors couple to distinct G proteins to effect stimulation of alveolar macrophage host defense functions. AB - Leukotrienes (LTs) are lipid mediators implicated in asthma and other inflammatory diseases. LTB(4) and LTD(4) also participate in antimicrobial defense by stimulating phagocyte functions via ligation of B leukotriene type 1 (BLT1) receptor and cysteinyl LT type 1 (cysLT1) receptor, respectively. Although both Galpha(i) and Galpha(q) proteins have been shown to be coupled to both BLT1 and cysLT1 receptors in transfected cell systems, there is little known about specific G protein subunit coupling to LT receptors, or to other G protein coupled receptors, in primary cells. In this study we sought to define the role of specific G proteins in pulmonary alveolar macrophage (AM) innate immune responses to LTB(4) and LTD(4). LTB(4) but not LTD(4) reduced cAMP levels in rat AM by a pertussis toxin (PTX)-sensitive mechanism. Enhancement of FcgammaR mediated phagocytosis and bacterial killing by LTB(4) was also PTX-sensitive, whereas that induced by LTD(4) was not. LTD(4) and LTB(4) induced Ca(2+) and intracellular inositol monophosphate accumulation, respectively, highlighting the role of Galpha(q) protein in mediating PTX-insensitive LTD(4) enhancement of phagocytosis and microbicidal activity. Studies with liposome-delivered G protein blocking Abs indicated a dependency on specific Galpha(q/11) and Galpha(i3) subunits, but not Galpha(i2) or G(beta)gamma, in LTB(4)-enhanced phagocytosis. The selective importance of Galpha(q/11) protein was also demonstrated in LTD(4) enhanced phagocytosis. The present investigation identifies differences in specific G protein subunit coupling to LT receptors in antimicrobial responses and highlights the importance of defining the specific G proteins coupled to heptahelical receptors in primary cells, rather than simply using heterologous expression systems. PMID- 17911633 TI - Identification of the IL-17 receptor related molecule IL-17RC as the receptor for IL-17F. AB - The proinflammatory cytokines IL-17A and IL-17F have a high degree of sequence similarity and share many biological properties. Both have been implicated as factors contributing to the progression of inflammatory and autoimmune diseases. Moreover, reagents that neutralize IL-17A significantly ameliorate disease severity in several mouse models of human disease. IL-17A mediates its effects through interaction with its cognate receptor, the IL-17 receptor (IL-17RA). We report here that the IL-17RA-related molecule, IL-17RC is the receptor for IL 17F. Notably, both IL-17A and IL-17F bind to IL-17RC with high affinity, leading us to suggest that a soluble form of this molecule may serve as an effective therapeutic antagonist of IL-17A and IL-17F. We generated a soluble form of IL 17RC and demonstrate that it effectively blocks binding of both IL-17A and IL 17F, and that it inhibits signaling in response to these cytokines. Collectively, our work indicates that IL-17RC functions as a receptor for both IL-17A and IL 17F and that a soluble version of this protein should be an effective antagonist of IL-17A and IL-17F mediated inflammatory diseases. PMID- 17911634 TI - The chemokine CCL6 promotes innate immunity via immune cell activation and recruitment. AB - Septic syndrome is a consequence of innate immune failure. Recent studies showed that the CC chemokine CCL6 enhanced antimicrobial immunity during experimental sepsis through an unknown mechanism. The present study demonstrates that transgenic CCL6 expression abolishes mortality in a septic peritonitis model via the modulation of resident peritoneal cell activation and, more importantly, through the recruitment of IFN-producing NK cells and killer dendritic cells into the peritoneum. Thus, CCL6 attenuates the immune failure during sepsis, in part, through a protective type 1-cytokine mediated mechanism. PMID- 17911635 TI - Adiponectin enhances IL-6 production in human synovial fibroblast via an AdipoR1 receptor, AMPK, p38, and NF-kappa B pathway. AB - Articular adipose tissue is a ubiquitous component of human joints, and adiponectin is a protein hormone secreted predominantly by differentiated adipocytes and involved in energy homeostasis. We investigated the signaling pathway involved in IL-6 production caused by adiponectin in both rheumatoid arthritis synovial fibroblasts and osteoarthritis synovial fibroblasts. Rheumatoid arthritis synovial fibroblasts and osteoarthritis synovial fibroblasts expressed the AdipoR1 and AdipoR2 isoforms of the adiponectin receptor. Adiponectin caused concentration- and time-dependent increases in IL-6 production. Adiponectin-mediated IL-6 production was attenuated by AdipoR1 and 5' AMP-activated protein kinase (AMPK)alpha1 small interference RNA. Pretreatment with AMPK inhibitor (araA and compound C), p38 inhibitor (SB203580), NF-kappaB inhibitor, IkappaB protease inhibitor, and NF-kappaB inhibitor peptide also inhibited the potentiating action of adiponectin. Adiponectin increased the kinase activity and phosphorylation of AMPK and p38. Stimulation of synovial fibroblasts with adiponectin activated IkappaB kinase alpha/beta (IKK alpha/beta), IkappaBalpha phosphorylation, IkappaBalpha degradation, p65 phosphorylation at Ser (276), p65 and p50 translocation from the cytosol to the nucleus, and kappaB-luciferase activity. Adiponectin-mediated an increase of IKK alpha/beta activity, kappaB-luciferase activity, and p65 and p50 binding to the NF-kappaB element and was inhibited by compound C, SB203580 and AdipoR1 small interference RNA. Our results suggest that adiponectin increased IL-6 production in synovial fibroblasts via the AdipoR1 receptor/AMPK/p38/IKKalphabeta and NF kappaB signaling pathway. PMID- 17911636 TI - Proteinase-activated receptor-2 exerts protective and pathogenic cell type specific effects in Alzheimer's disease. AB - The proteinase-activated receptors (PARs) are a novel family of G protein-coupled receptors, and their effects in neurodegenerative diseases remain uncertain. Alzheimer's disease (AD) is a neurodegenerative disorder defined by misfolded protein accumulation with concurrent neuroinflammation and neuronal death. We report suppression of proteinase-activated receptor-2 (PAR2) expression in neurons of brains from AD patients, whereas PAR2 expression was increased in proximate glial cells, together with up-regulation of proinflammatory cytokines and chemokines and reduced IL-4 expression (p < 0.05). Glial PAR2 activation increased expression of formyl peptide receptor-2 (p < 0.01), a cognate receptor for a fibrillar 42-aa form of beta-amyloid (Abeta(1-42)), enhanced microglia mediated proinflammatory responses, and suppressed astrocytic IL-4 expression, resulting in neuronal death (p < 0.05). Conversely, neuronal PAR2 activation protected human neurons against the toxic effects of Abeta(1-42) (p < 0.05), a key component of AD neuropathogenesis. Amyloid precursor protein-transgenic mice, displayed glial fibrillary acidic protein and IL-4 induction (p < 0.05) in the absence of proinflammatory gene up-regulation and neuronal injury, whereas PAR2 was up-regulated at this early stage of disease progression. PAR2-deficient mice, after hippocampal Abeta(1-42) implantation, exhibited enhanced IL-4 induction and less neuroinflammation (p < 0.05), together with improved neurobehavioral outcomes (p < 0.05). Thus, PAR2 exerted protective properties in neurons, but its activation in glia was pathogenic with secretion of neurotoxic factors and suppression of astrocytic anti-inflammatory mechanisms contributing to Abeta(1 42)-mediated neurodegeneration. PMID- 17911637 TI - Hepatocyte growth factor significantly suppresses collagen-induced arthritis in mice. AB - Hepatocyte growth factor (HGF) plays an important role in angiogenesis, cell proliferation, antifibrosis, and antiapoptosis. Moreover, recent studies have highlighted the immunosuppressive effect of HGF in animal models of allogenic heart transplantation and autoimmune myocarditis and in studies in vitro as well. We also reported that HGF significantly suppresses dendritic cell function, thus down-regulating Ag-induced Th1-type and Th2-type immune responses in allergic airway inflammation. However, the immunosuppressive effect of HGF in many other situations has not been fully clarified. In the present study, using a mouse model of collagen-induced arthritis (CIA) and experiments in vitro, we examined the effect of HGF on autoimmune arthritis and then elucidated the mechanisms of action of HGF. To achieve sufficient delivery of HGF, we used biodegradable gelatin hydrogels as a carrier. HGF suppressed Ag-induced T cell priming by regulating the functions of dendritic cells in the Ag-sensitization phase with down-regulation of IL-10. In contrast, under continuous Ag stimulation HGF induced IL-10-producing immunocytes both in vivo and in vitro. Moreover, HGF potently inhibited the development of CIA with enhancing the Th2-type immune response. We also confirmed that HGF significantly suppressed the production of IL-17 by immunocytes. These results indicate that HGF suppresses the development of CIA through different ways at different phases. They also suggest that HGF could be an attractive tool for treating patients with rheumatoid arthritis. PMID- 17911638 TI - The IFN-inducible GTPase LRG47 (Irgm1) negatively regulates TLR4-triggered proinflammatory cytokine production and prevents endotoxemia. AB - LRG47/Irgm1, a 47-kDa IFN-inducible GTPase, plays a major role in regulating host resistance as well as the hemopoietic response to intracellular pathogens. LRG47 expression in macrophages has been shown previously to be stimulated in vitro by bacterial LPS, a TLR4 ligand. In this study, we demonstrate that induction of LRG47 by LPS is not dependent on MyD88 signaling, but rather, requires STAT-1 and IFN-beta. In addition, LRG47-deficient mice are highly susceptible to LPS, but not TLR2 ligand-induced shock, an outcome that correlates with enhanced proinflammatory cytokine production in vitro and in vivo. Further analysis revealed that LPS-stimulated LRG47-deficient macrophages display enhanced phosphorylation of p38, a downstream response associated with TLR4/MyD88 rather than IFN-beta/STAT-1 signaling. In contrast, LPS-induced phosphorylation of IFN regulatory factor-3 and expression of IFN-beta or the type I IFN-regulated genes, CCL5 and CCL10, were unaltered in LRG47(-/-) cells. Together, these observations indicate that in LPS-stimulated murine macrophages LRG47 is induced by IFN-beta and negatively regulates TLR4 signaling to prevent excess proinflammatory cytokine production and shock. Thus, our findings reveal a new host-protective function for this GTPase in the response to pathogenic encounter. PMID- 17911639 TI - The heat shock protein HSP70 promotes mouse NK cell activity against tumors that express inducible NKG2D ligands. AB - The stress-inducible heat shock protein (HSP) 70 is known to function as an endogenous danger signal that can increase the immunogenicity of tumors and induce CTL responses. We show in this study that HSP70 also activates mouse NK cells that recognize stress-inducible NKG2D ligands on tumor cells. Tumor size and the rate of metastases derived from HSP70-overexpressing human melanoma cells were found to be reduced in T and B cell-deficient SCID mice, but not in SCID/beige mice that lack additionally functional NK cells. In the SCID mice with HSP70-overexpressing tumors, NK cells were activated so that they killed ex vivo tumor cells that expressed NKG2D ligands. In the tumors, the MHC class I chain related (MIC) A and B molecules were found to be expressed. Interestingly, a counter selection was observed against the expression of MICA/B in HSP70 overexpressing tumors compared with control tumors in SCID, but not in SCID/beige mice, suggesting a functional relevance of MICA/B expression. The melanoma cells were found to release exosomes. HSP70-positive exosomes from the HSP70 overexpressing cells, in contrast to HSP70-negative exosomes from the control cells, were able to activate mouse NK cells in vitro to kill YAC-1 cells, which express NKG2D ligands constitutively, or the human melanoma cells, in which MICA/B expression was induced. Thus, HSP70 and inducible NKG2D ligands synergistically promote the activation of mouse NK cells resulting in a reduced tumor growth and suppression of metastatic disease. PMID- 17911640 TI - Dual T cell receptor expressing CD8+ T cells with tumor- and self-specificity can inhibit tumor growth without causing severe autoimmunity. AB - The engineering of Ag-specific T cells by expression of TCR genes is a convenient method for adoptive T cell immunotherapy. A potential problem is the TCR gene transfer into self-reactive T cells that survived tolerance mechanisms. We have developed an experimental system with T cells that express two TCRs with defined Ag-specificities, one recognizing a tumor-specific Ag (LCMV-gp(33)), the other recognizing a self-Ag in the pancreas (OVA). By using tumor cells expressing high and low amounts of Ag and mice expressing high and low levels of self-Ag in the pancreas (RIP-OVA-Hi and RIP-OVA-Lo), we show that 1) tumor rejection requires high amount of tumor Ag, 2) severe autoimmunity requires high amount of self-Ag, and 3) if Ag expression on tumor cells is sufficient and low in the pancreas, successful adoptive T cell therapy can be obtained in the absence of severe autoimmunity. These results are shown with T cells from dual TCR transgenic mice or T cells that were redirected by TCR gene transfer. Our data demonstrate that the approach of adoptively transferring TCR redirected T cells can be effective without severe side effects, even when high numbers of T cells with self reactivity were transferred. PMID- 17911641 TI - Hematin promotes complement alternative pathway-mediated deposition of C3 activation fragments on human erythrocytes: potential implications for the pathogenesis of anemia in malaria. AB - Childhood malaria caused by Plasmodium falciparum is often characterized by severe anemia at low parasite burdens; the mechanism(s) responsible for this pathology remain to be defined. We have reported, based on clinical observations and in vitro models, that complement control proteins on erythrocytes such as CR1, the immune adherence receptor specific for C3b, may be reduced in childhood malaria, suggesting a possible role for complement in erythrocyte destruction. Intravascular lysis of iE by P. falciparum leads to release of erythrocyte breakdown products such as hemoglobin and hematin, which have inflammatory properties. In the present article, we demonstrate that in serum and in anticoagulated whole blood, moderate concentrations of hematin activate the alternative pathway of complement and promote deposition of C3 activation and breakdown products on erythrocytes. The degree of C3 fragment deposition is directly correlated with erythrocyte CR1 levels, and erythrocytes opsonized with large amounts of C3dg form rosettes with Raji cells, which express CR2, the C3dg receptor which is expressed on several types of B cells in the spleen. Thus, the reaction mediated by hematin promotes opsonization and possible clearance of the youngest (highest CR1) erythrocytes. A mAb specific for C3b, previously demonstrated to inhibit the alternative pathway of complement, completely blocks the C3 fragment deposition reaction. Use of this mAb in nonhuman primate models of malaria may provide insight into mechanisms of erythrocyte destruction and thus aid in the development of targeted therapies based on inhibiting the alternative pathway of complement. PMID- 17911642 TI - Impaired T cell protein kinase C delta activation decreases ERK pathway signaling in idiopathic and hydralazine-induced lupus. AB - T cells from patients with lupus or treated with the lupus-inducing drug hydralazine have defective ERK phosphorylation. The reason for the impaired signal transduction is unknown but important to elucidate, because decreased T cell ERK pathway signaling causes a lupus-like disease in animal models by decreasing DNA methyltransferase expression, leading to DNA hypomethylation and overexpression of methylation-sensitive genes with subsequent autoreactivity and autoimmunity. We therefore analyzed the PMA stimulated ERK pathway phosphorylation cascade in CD4(+) T cells from patients with lupus and in hydralazine-treated cells. The defect in these cells localized to protein kinase C (PKC)delta. Pharmacologic inhibition of PKCdelta or transfection with a dominant negative PKCdelta mutant caused demethylation of the TNFSF7 (CD70) promoter and CD70 overexpression similar to lupus and hydralazine-treated T cells. These results suggest that defective T cell PKCdelta activation may contribute to the development of idiopathic and hydralazine-induced lupus through effects on T cell DNA methylation. PMID- 17911643 TI - Mature B cells preferentially lose tolerance in the chronic graft-versus-host disease model of systemic lupus erythematosus. AB - Chronic graft-vs-host (cGVH) disease is a well-characterized systemic lupus erythematosus (SLE) model. Induction of cGVH in anti-DNA H chain knockin (3H9KI) transgenic mice results in specific activation of anti-dsDNA B cells. In this study, we show that B cells from 3H9KI mice were activated by cGVH even when adoptively transferred into irradiated JHT-/- recipients that lack endogenous B cells. This process of activation was reflected by high autoantibody titers and changes in phenotypic markers. We have used this system to characterize the particular B cell subsets that were responsible for secreting autoantibodies during cGVH response. We isolated splenic B cell subsets based on their expression of specific cell surface markers and used them in our adoptive transfer studies. We found that mature B cells were the most vulnerable to the allostimulus and were the major source of autoantibodies compared with immature B cells. The greater susceptibility of mature B cells to become activated and thereby lose tolerance was unanticipated and has implications for maintenance of peripheral tolerance and for the development of autoimmunity. Furthermore, of the mature B cells, marginal zone B cells were particularly responsible for mounting the initial response to the cGVH stimulus. This observation underscores the critical role of marginal zone B cells in activation and production of autoantibodies. PMID- 17911644 TI - Intravenous immunoglobulin therapy affects T regulatory cells by increasing their suppressive function. AB - Intravenous Ig therapy (IVIg) is reported to be a useful regimen in treating autoimmune diseases. In this study, we asked whether IVIg (in vitro) could increase the expression of TGF-beta, IL-10, and the transcription factor FoxP3 in T regulatory (Treg) cells, and the idea that IVIg could enhance suppressive properties of these cells. CD4(+) T cells from 12 healthy individuals were cultured in the presence or absence of IVIg vs human control IgG during 16, 24, and 36 h. Using FACS analysis and gating on CD4(+)CD25(high) Treg cells, we assessed the expression of intracellular TGF-beta, IL-10, and FoxP3. In addition, the production of TNF-alpha by stimulated CD4(+) T cells alone or in culture with CD25(+) by itself or together with IVIg was also assessed. The presence of IVIg with Treg cells in culture significantly increased the intracellular expression of TGF-beta (17.7 +/- 8.5% vs 29.8 +/- 13%; p = 0.02), IL-10 (20.7 +/- 4.7% vs 34.2 +/- 5.2%; p = 0.008) and FoxP3 (20.8 +/- 5.2% vs 33.7 +/- 5.9%; p = 0.0006) when compared with cells cultured alone or with control human IgG. The suppressive effect of CD4(+)CD25(+) T cells presented as the decrease of TNF alpha production by stimulated CD4(+)CD25(-) (effector T cells) was further increased by adding IVIg to cell culture. We hereby demonstrate an additional mechanism by which IVIg could maintain self-tolerance and decrease immune mediated inflammation. PMID- 17911645 TI - Exacerbation of collagen-induced arthritis by oligoclonal, IL-17-producing gamma delta T cells. AB - Murine gammadelta T cell subsets, defined by their Vgamma chain usage, have been shown in various disease models to have distinct functional roles. In this study, we examined the responses of the two main peripheral gammadelta T cell subsets, Vgamma1(+) and Vgamma4(+) cells, during collagen-induced arthritis (CIA), a mouse model that shares many hallmarks with human rheumatoid arthritis. We found that whereas both subsets increased in number, only the Vgamma4(+) cells became activated. Surprisingly, these Vgamma4(+) cells appeared to be Ag selected, based on preferential Vgamma4/Vdelta4 pairing and very limited TCR junctions. Furthermore, in both the draining lymph node and the joints, the vast majority of the Vgamma4/Vdelta4(+) cells produced IL-17, a cytokine that appears to be key in the development of CIA. In fact, the number of IL-17-producing Vgamma4(+) gammadelta T cells in the draining lymph nodes was found to be equivalent to the number of CD4(+)alphabeta(+) Th-17 cells. When mice were depleted of Vgamma4(+) cells, clinical disease scores were significantly reduced and the incidence of disease was lowered. A decrease in total IgG and IgG2a anti-collagen Abs was also seen. These results suggest that Vgamma4/Vdelta4(+) gammadelta T cells exacerbate CIA through their production of IL-17. PMID- 17911646 TI - OX40 controls functionally different T cell subsets and their resistance to depletion therapy. AB - T cell depletion is a widely used approach in clinical transplantation. However, not all T cells are equally sensitive to depletion therapies and a significant fraction of T cells persists even after aggressive treatment. The functional attributes of such T cells and the mechanisms responsible for their resistance to depletion are poorly studied. In the present study, we showed that CD4(+) T cells that are resistant to polyclonal anti-lymphocyte serum (ALS) mediated depletion exhibit phenotypic features of memory cells and uniformly express OX40 on the cell surface. Studies using the foxp3gfp knockin mice revealed that the remaining CD4(+)OX40(+) cells consist of Foxp3(+) Tregs and Foxp3(-) T effector/memory cells. The ALS-resistant CD4(+)OX40(+) cells failed to mediate skin allograft rejection upon adoptive transferring into congenic Rag(-/-) mice, but removal of Foxp3(+) Tregs from the OX40(+) cells resulted in prompt skin allograft rejection. Importantly, OX40 is critical to survival of both Foxp3(+) Tregs and T effector/memory cells. However, OX40 exhibits opposing effects on the functional status of Foxp3(+) Tregs and T effector/memory cells, as stimulation of OX40 on T effector cells induced amplified cell proliferation but stimulation of OX40 on the Foxp3(+) Tregs impaired their suppressor functions. Our study demonstrates that OX40 is a critical molecule in regulating survival and functions of depletion-resistant T cells; and these findings may have important clinical implications. PMID- 17911647 TI - IL-10- and TGF-beta-mediated susceptibility in kala-azar and post-kala-azar dermal leishmaniasis: the significance of amphotericin B in the control of Leishmania donovani infection in India. AB - Visceral leishmaniasis (VL) or kala-azar is known to be associated with a mixed Th1-Th2 response, and effective host defense requires the induction of IFN-gamma and IL-12. We address the role of the differential decline of IL-10 and TGF-beta in response to sodium antimony gluconate (SAG) and amphotericin B (AmB), the therapeutic success of SAG and AmB in Indian VL, and the significance of IL-10 and TGF-beta in the development and progression of post-kazla-azar dermal leishmaniasis (PKDL). In the active disease, PBMC from VL patients showed suppressed Ag-specific lymphoproliferation, IFN-gamma and IL-12 production, and elevation of IL-10 and TGF-beta. Cure corresponded with an elevation in IFN-gamma and IL-12 production and down-regulation of IL-10 and TGF-beta. Both CD4(+) and CD8(+) T cells were involved in IFN-gamma and IL-10 production. Interestingly, the retention and maintenance of residual IL-10 and TGF-beta in some SAG-treated individuals and the elevation of IL-10 and TGF-beta in PKDL, a sequel to kala azar, probably reflects the role of these cytokines in reactivation of the disease in the form of PKDL. Contrastingly, AmB treatment of VL resulted in negligible TGF-beta levels and absolute elimination of IL-10, reflecting the better therapeutic activity of AmB and its probable role in the recent decline in PKDL occurrences in India. Moreover, elucidation of immune responses in Indian PKDL patients revealed a spectral pattern of disease progression where disease severity could be correlated inversely with lymphoproliferation and directly with TGF-beta, IL-10, and Ab production. In addition, the enhancement of CD4(+)CD25(+) T cells in active VL, their decline at cure, and reactivation in PKDL suggest their probable immunosuppressive role in these disease forms. PMID- 17911649 TI - Pollen lethality: a phenomenon in Arabidopsis RNA interference plants. PMID- 17911648 TI - Molecular dissection of endosomal compartments in plants. PMID- 17911650 TI - Nonmotile cellulose synthase subunits repeatedly accumulate within localized regions at the plasma membrane in Arabidopsis hypocotyl cells following 2,6 dichlorobenzonitrile treatment. PMID- 17911651 TI - Gene-hormone-environment interactions in the regulation of aggressive responses: elegant analysis of complex behavior. AB - Photoperiodic influences on the relations between levels of a steroid hormone and aggressive behavior add a layer of complexity to our ideas about how genes influence behavior. The effects of estrogens on aggression are modulated by day length and involve both genomic and nongenomic routes of hormone action. PMID- 17911652 TI - Studying integrin-mediated cell adhesion at the single-molecule level using AFM force spectroscopy. AB - The establishment of cell adhesion involves specific recognition events between individual cell-surface receptors and molecules of the cellular environment. However, characterizing single-molecule adhesion events in the context of a living cell presents an experimental challenge. The atomic force microscope (AFM) operated in force spectroscopy mode provides an ultrasensitive method to investigate cell adhesion forces at the level of single receptor-ligand bonds. With a living cell attached to the AFM cantilever, the number of cell-substrate interactions can be controlled and limited to the formation of single receptor ligand bonds. From force-distance (F-D) curves recorded during cell detachment, the strength of single receptor-ligand bonds can be determined. Furthermore, by varying the rate of force application during bond rupture, a dynamic force spectrum (DFS) can be generated from which additional parameters that describe the energy landscape of the interaction, such as dissociation rate and energy barrier width, can be obtained. Using the example of alpha(2)beta(1) integrin mediated adhesion to type I collagen, we provide a detailed description of how dynamic AFM single-cell force spectroscopy (SCFS) adhesion measurements can be performed with single-molecule sensitivity, and how specific energy landscape parameters of the integrin-collagen bond can be extracted from the DFS. PMID- 17911653 TI - The T/G 13915 variant upstream of the lactase gene (LCT) is the founder allele of lactase persistence in an urban Saudi population. AB - BACKGROUND: The prevalence of lactase persistence is high in Saudi Arabia. OBJECTIVE: To identify a DNA variant for the lactase persistence/non-persistence trait in adult Arabs in Saudi Arabia. METHODS: We sequenced DNA from 432 anonymous neonatal blood donors from five different regions of Saudi Arabia to cover the 400 bp region surrounding the previously identified lactase persistence/non-persistence variant C/T-13910 residing in intron 13 of the MCM6 gene. RESULTS: Two anonymous blood donors carried the C/T-13910 genotype. One variant, T/G -13915, residing 5 bp upstream of the C/T-13910 variant, was present in 332 of 432 (76.9%) of the neonatal samples, compatible with previous prevalence figures of lactase persistence in urban Saudi populations. Determination of disaccharidase activities in 25 intestinal biopsy samples showed a highly significant correlation between lactase activity and the T/G-13915 genotypes (p<0.001; Fisher exact test) as well as between the L:S ratio and the aforementioned genotypes (p<0.001; Fisher exact test). CONCLUSION: The T/G-13915 variant is the founder mutation of lactase persistence in an urban Saudi population. The results obtained here have implications for genetic testing of adult-type hypolactasia and to analysis of human evolution, the origin of cattle domestication and migrations of the populations in the Arabian peninsula. PMID- 17911654 TI - Anthropometric evaluation of children with SHOX mutations can be used as indication for genetic studies in children of short stature. PMID- 17911656 TI - Deletion of a 760 kb region at 4p16 determines the prenatal and postnatal growth retardation characteristic of Wolf-Hirschhorn syndrome. AB - BACKGROUND: Recently the genotype/phenotype map of Wolf-Hirschhorn syndrome (WHS) has been refined, using small 4p deletions covering or flanking the critical region in patients showing only some of the WHS malformations. Accordingly, prenatal-onset growth retardation and failure to thrive have been found to result from haploinsufficiency for a 4p gene located between 0.4 and 1.3 Mb, whereas microcephaly results from haploinsufficiency of at least two different 4p regions, one of 2.2-2.38 Mb and a second one of 1.9-1.28 Mb. METHODS AND RESULTS: We defined the deletion size of a ring chromosome (r(4)) in a girl with prenatal onset growth retardation, severe failure to thrive and true microcephaly but without the WHS facial gestalt and mental retardation. A high-resolution comparative genome hybridisation array revealed a 760 kb 4p terminal deletion. CONCLUSIONS: This case, together with a familial 4p deletion involving the distal 400 kb reported in normal women, may narrow the critical region for short stature on 4p to 360-760 kb. This region is also likely to contain a gene for microcephaly. "In silico" analysis of all genes within the critical region failed to reveal any strikingly suggestive expression pattern; all genes remain candidates for short stature and microcephaly. PMID- 17911657 TI - Fluoro-edenite fibers induce expression of Hsp70 and inflammatory response. AB - Many asbestos-like mineral fibers have been detected in the air of mountainous and volcanic areas of Italy and other parts of the world. These fibers have been suspected to be the cause of increased incidences of lung cancer and other lung diseases in these areas. However, the mechanisms of the cellular response and defense following exposure to these microscopic fibers have not been characterized. We continue to study these mechanisms to be able to propose preventive strategies in large populations. The objective of the present study was to determine comparatively biological responses of mesothelial Met-5A and monocyte-macrophage J774 cells following exposure to two types of fluoro-edenite fibers having low and high iron content (labeled 19 and 27, respectively) obtained from Biancavilla (Sicily, Italy). The reference fiber was a non-iron fibrous tremolite from Val di Susa (Piemonte, Italy). The cells were treated with 5, 50, and 100 mug of fibrous matter per 1 ml for 72 hr. We identified several key mechanisms by which cells responded and counteracted the injury induced by these fibers. The fibers caused induction of the heat shock protein 70 (Hsp70), stimulated formation of reactive oxygen species (detected by using DCFH-DA as a fluorescent probe) and NO* (measured as nitrite). Exposure of cells to the fibers induced lactate dehydrogenase activity and decreased viability. The fluoro endenite type 27 was the most potent fiber tested, which indicated that iron and possibly manganese contribute significantly to this fiber toxicity. The J774 cells were more sensitive to fluoro-edenite than Met-5A cells suggesting that the primary site of the fiber-induced inflammatory response could be the macrophage rather than the pulmonary epithelium. Fluoro-edenite produces more biological alterations with respect to non-iron tremolite. Hsp70 and free radicals could be important factors in the context of mineral fiber-induced acute lung injury leading possibly to mutagenic effects. We anticipate that pharmacological blockade of the fiber-dependent cellular responses could in long term offer preventive approach to combat lung diseases induced by these fibers. PMID- 17911659 TI - Dose-related increased binding of nickel to chromatin proteins; and changes to DNA concentration in the liver of guinea pigs treated with Nigerian light crude oil. AB - The alteration in nuclear DNA concentration and the concomitant binding of xenobiotics (alkylating agents, heavy metals, etc.) to chromatin constituents may adversely affect gene structure and/or function, and thus initiate carcinogenesis. Binding of nickel to chromatin DNA has been reported to cause DNA damage (cross-links, single-strand breaks), and although many soluble nickel compounds and complexes have been shown to bind to chromatin, porphyrin-complexed nickel (PCN) in crude oils has not been studied. We have determined the doserelated increases in total and chromatin DNA concentrations, and the differential distribution (binding) of PCN (crude oil nickel-CON) to chromatin constituents in livers of adult male guinea pigs treated with 1.25, 2.50 and 5.0 ml/kg bw Nigerian Bonny light crude oil (BLCO) by intraperitoneal injection. The results showed large BLCO-induced increases in total DNA concentrations of 424%, 632% and 436% at 1.25, 2.50 and 5.0 ml/kg bw BLCO respectively over the untreated controls; while it induced equally large increases in chromatin DNA concentrations of 585% and 200% at 2.50 and 5.0 ml/kg bw respectively. In both cases, maximum increases occurred at 2.50 ml/kg bw BLCO. The distribution of PCN in BLCO between chromatin DNA and chromatin proteins (histones and non-histones) showed that at 2.50 and 5.0 ml/kg bw BLCO, nickel content in chromatin DNA reduced by 25% and 12.5% respectively over the controls; while its content in chromatin proteins also reduced by 26%; but increased by 166% at 2.50 and 5.0 ml/kg bw BLCO, respectively over the untreated controls. However, in intra chromatin comparison, 38.8% more PCN bound to chromatin DNA than to chromatin proteins at 2.50 ml/kg bw; but at 5.0 ml/kg bw BLCO, 90.4% more PCN bound to chromatin proteins than to chromatin DNA. These results show a greater affinity of PCN in BLCO for chromatin proteins over chromatin DNA which may have played a role in the increased DNA concentrations. Also, the results may add critical information to understanding the reactions of porphyrin-complexed nickel in crude oils with chromatin since this has not been studied before. Furthermore, the probable carcinogenicity of BLCO may be implied. PMID- 17911658 TI - Mathematical models of cobalt and iron ions catalyzed microwave bacterial deactivation. AB - Time differences for Enterococcus faecalis, Staphylococcus aureus, and Escherichia coli survival during microwave irradiation (power 130 W) in the presence of aqueous cobalt and iron ions were investigated. Measured dependencies had "bell" shape forms with maximum bacterial viability between 1 - 2 min becoming insignificant at 3 minutes. The deactivation time for E. faecalis, S. aureus and E.coli in the presence of metal ions were smaller compared to a water control (4 -5 min). Although various sensitivities to the metal ions were observed, S. aureus and E. coli and were the most sensitive for cobalt and iron, respectively. The rapid reduction of viable bacteria during microwave treatment in the presence of metal ions could be explained by increased metal ion penetration into bacteria. Additionally, microwave irradiation may have increased the kinetic energy of the metal ions resulting in lower survival rates. The proposed mathematical model for microwave heating took into account the "growth" and "death" factors of the bacteria, forming second degree polynomial functions. Good relationships were found between the proposed mathematical models and the experimental data for bacterial deactivation (coefficient of correlation 0.91 - 0.99). PMID- 17911661 TI - Effect of arsenic and chromium on the serum amino-transferases activity in Indian major carp, Labeo rohita. AB - Arsenic and hexavalent chromium toxicity results from their ability to interact with sulfahydryl groups of proteins and enzymes, and to substitute phosphorus in a variety of biochemical reactions. Alanine aminotransferase (ALT; E.C: 2.6.1.2) and Aspartate amino transferase (AST; EC 2.6.1.1) play a crucial role in transamination reactions and can be used as potential biomarkers to indicate hepatotoxicity and cellular damage. While histopathological studies in liver tissue require more time and expertise, simple and reliable biochemical analysis of ALT and AST can be used for a rapid assessment of tissue and cellular damage within 96 h. The main objective of this study was to determine the acute effects of arsenic and hexavalent chromium on the activity of ALT and AST in the Indian major carp, Labeo rohita for 24 h and 96 h. Significant increase in the activity of ALT (P < 0.01) from controls in arsenic exposed fish indicates serious hepatic damage and distress condition to the fish. However, no such significant changes were observed in chromium-exposed fish suggesting that arsenic is more toxic to the fish. These findings indicate that ALT and AST are candidate biomarkers for arsenic-induced hepatotoxicity in Labeo rohita. PMID- 17911660 TI - Genotoxicity of air borne particulates assessed by comet and the Salmonella mutagenicity test in Jeddah, Saudi Arabia. AB - Fine airborne respirable particulates less than 10 micrometer (PM10) are considered one of the top environmental public health concerns, since they contain polycyclic aromatic hydrocarbons (PAHs) which are among the major carcinogenic compounds found in urban air. The objective of this study is to assess the genotoxicity of the ambient PM10 collected at 11 urban sites in Jeddah, Saudi Arabia. The PM10 extractable organic matter (EOM) was examined for its genotoxicity by the single cell gel electrophoresis (SCGE) comet assay and the Salmonella mutagenicity (Ames) test .Gas chromatography-mass spectrometry was used to quantify 16 PAH compounds in four sites. Samples from oil refinery and heavy diesel vehicles traffic sites showed significant DNA damage causing comet in 20-44% of the cells with tail moments ranging from 0.5-2.0 compared to samples from petrol driven cars and residential area, with comet in less than 2% of the cells and tail moments of < 0.02. In the Ames test, polluted sites showed indirect mutagenic response and caused 20-56 rev/ m3, mean while residential and reference sites caused 2-15 rev /m3. The genotoxicity of the EOM in both tests directly correlated with the amount of organic particulate and the PAHs concentrations in the air samples. The PAHs concentrations ranged between 0.83 ng/m3 in industrial and heavy diesel vehicles traffic sites to 0.18 ng /m3 in the residential area. Benzo(ghi)pyrene was the major PAH components and at one site it represented 65.4 % of the total PAHs. Samples of the oil refinery site were more genotoxic in the SCGE assay than samples from the heavy diesel vehicles traffic site, despite the fact that both sites contain almost similar amount of PAHs. The opposite was true for the mutagenicity in the Ames test. This could be due to the nature of the EOM in both sites. These findings confirm the genotoxic potency of the PM10 organic extracts to which urban populations are exposed. PMID- 17911662 TI - Evaluation of HACCP plans of food industries: case study conducted by the Servizio di Igiene degli Alimenti e della Nutrizione (Food and Nutrition Health Service) of the local health authority of Foggia, Italy. AB - With respect to food safety, many works have studied the effectiveness of self monitoring plans of food companies, designed using the Hazard Analysis and Critical Control Point (HACCP) method. On the other hand, indepth research has not been made concerning the adherence of the plans to HACCP standards. During our research, we evaluated 116 self-monitoring plans adopted by food companies located in the territory of the Local Health Authority (LHA) of Foggia, Italy. The general errors (terminology, philosophy and redundancy) and the specific errors (transversal plan, critical limits, hazard specificity, and lack of procedures) were standardized. Concerning the general errors, terminological errors pertain to half the plans examined, 47% include superfluous elements and 60% have repetitive subjects. With regards to the specific errors, 77% of the plans examined contained specific errors. The evaluation has pointed out the lack of comprehension of the HACCP system by the food companies and has allowed the Servizio di Igiene degli Alimenti e della Nutrizione (Food and Nutrition Health Service), in its capacity as a control body, to intervene with the companies in order to improve designing HACCP plans. PMID- 17911663 TI - Mapping cigarettes similarities using cluster analysis methods. AB - The aim of the research was to investigate the relationship and/or occurrences in and between chemical composition information (tar, nicotine, carbon monoxide), market information (brand, manufacturer, price), and public health information (class, health warning) as well as clustering of a sample of cigarette data. A number of thirty cigarette brands have been analyzed. Six categorical (cigarette brand, manufacturer, health warnings, class) and four continuous (tar, nicotine, carbon monoxide concentrations and package price) variables were collected for investigation of chemical composition, market information and public health information. Multiple linear regression and two clusterization techniques have been applied. The study revealed interesting remarks. The carbon monoxide concentration proved to be linked with tar and nicotine concentration. The applied clusterization methods identified groups of cigarette brands that shown similar characteristics. The tar and carbon monoxide concentrations were the main criteria used in clusterization. An analysis of a largest sample could reveal more relevant and useful information regarding the similarities between cigarette brands. PMID- 17911664 TI - Qat habit in Yemen society: a causative factor for oral periodontal diseases. AB - The effect of a common habit among Yemeni population on the periodontal status was investigated. This cross-sectional study was done on 2500 Yemenis with mean age 27.01 years (1818 males and 682 females). Among these 1528 were qat chewers and 972 were non-chewers. Detailed questionnaire and pre-designed scoring system for the periodontal status were employed for each case. Study results indicated that out of 972 non-chewers 116(12%) had periodontal pocketing and 18 (1.9%) cases had gingival recession. On the other hand, out of 1528 chewers, 468 (31.8%) had periodontal pockets and 98 (6.4%) with gum bleeding, p<0.05. These effects were found to increase with increased frequency and duration of chewing. It was concluded that habit of qat can cause damage to the periodontal ligament as pocketing and gum recession. PMID- 17911665 TI - HIV/AIDS knowledge, attitudes and behaviors assessment of Chinese students: a questionnaire study. AB - The objective of this study was to assess students' knowledge, attitudes and practices on HIV and AIDS. A questionnaire was administered to a cross section of 259 Chinese undergraduates. Respondents were asked to provide information about knowledge and attitudes about HIV/AIDS. Study results indicated that the majority of undergraduates had a moderate level of HIV and AIDS knowledge, acceptance and attitudes towards people with HIV and AIDS. Boys had more acceptance and positive attitudes towards people with HIV and AIDS than girls. Students majoring in medicine performed better (more knowledgeable and accepting) than non-medical students. Differences between students with various monthly expenditures were found-- 6.2% of students had 3-5 sexual partners which has rarely been found in Chinese students; most students did not know HIV VCT centers and most students did not show their confidence for controlling of HIV and AIDS in China. In conclusion, students' knowledge about HIV/AIDS was uneven. A peer educational program to talk about self esteem, healthy sexual attitudes, being human accepting and loving should be developed in the near future. PMID- 17911666 TI - Evaluation of the effect of a health education campaign of HIV by using an analytical hierarchy process method. AB - This study was designed to understand the status of HIV/AIDS knowledge, attitude and practice (KAP) among different populations and to provide scientific evidences for further health education. Three rounds of questionnaires were administered among service industry workers who were selected through stratified cluster sampling. Study subjects included hotel attendants, employees of beauty parlors and service workers of transportation industry. Data were analyzed using the analytical hierarchy process. All demonstrated high KAP overall. Synthetic scoring indexes of the three surveys were above 75%. However, the correct response rate on questions whether mosquito bite can transmit HIV/AIDS and what is the relationship between STD with HIV was unsatisfactory (lower than expected); and their attitudes towards people living with HIV and AIDS need to be improved. Moreover, the effect of health education on these groups was unclear. In conclusion, analytical hierarchy process is a valid method in estimating overall effect of HIV/AIDS health education. Although the present status of HIV/AIDS KAP among the service industry workers was relatively good, greater efforts should be made to improve their HIV transmission knowledge, attitude and understanding of the relationship between STDs and HIV. PMID- 17911667 TI - Linkcare--enabling continuity of care for the chronically ill across levels and profession. AB - Chronic diseases are increasing rapidly and this phenomenon is becoming a major burden to the health delivery system around the world. A new health care paradigm with focus on chronic treatment and care will actualize the need for interoperable standards based services due to the complexity of care where different health levels and professions are involved. Given the complexity of the domain, we argue the need for a systematic and formal approach to the development of interoperable information systems if there shall be any real support of the cooperating actors. We describe our work on technical interoperability done in the Linkcare project addressing new models of care and technology to support them in the domain of the chronically ill using concrete results from an architecture built using the MAFIIA architecture framework and the UML 2.0 profile for software services, and argue that building formal architectural descriptions on the basis of shared interface descriptions and profiles are an important part of achieving continuity of care based on sustainable health systems. PMID- 17911668 TI - Sustainable ubiquitous home health care--architectural considerations and first practical experiences. AB - Despite the abundance of past home care projects and the maturity of the technologies used, there is no widespread dissemination as yet. The absence of accepted standards and thus interoperability and the inadequate integration into transinstitutional health information systems (tHIS) are perceived as key factors. Based on the respective literature and previous experiences in home care projects we propose an architectural model for home care as part of a transinstitutional health information system using the HL7 clinical document architecture (CDA) as well as the HL7 Arden Syntax for Medical Logic Systems. In two short case studies we describe the practical realization of the architecture as well as first experiences. Our work can be regarded as a first step towards an interoperable - and in our view sustainable - home care architecture based on a prominent document standard from the health information system domain. PMID- 17911669 TI - Do physicians take action on high risk family history information provided by patients outside of a clinic visit? AB - Clinically relevant family history information is frequently missing or not readily available in electronic health records. Improving the availability of family history information is important for optimum care of many patients. Family history information on five conditions was collected in a survey from 163 primary care patients. Overall, 53% of patients had no family history information in the electronic health record (EHR) either on the patient's problem list or within a templated family history note. New information provided by patients resulted in an increase in the patient's risk level for 32% of patients with a positive family history of breast cancer, 40% for coronary artery disease, 50% for colon cancer, 74% for diabetes, and 95% each for osteoporosis and glaucoma. Informing physicians of new family history information outside of a clinic visit through an electronic clinical message and note in the EHR was not sufficient to achieve recommended follow-up care. Better tools need to be developed to facilitate the collection of family history information and to support clinical decision-making and action. PMID- 17911670 TI - Sharing electronic laboratory results in a patient portal--a feasibility pilot. AB - Surveys of patients consistently demonstrate a very strong interest in having secure electronic access to their own laboratory test results. In recent years, a number of health care providers and lab service centers have offered this capability, which now extends to millions of patients in the United States. Yet, little has been published on the methods of making lab results available. This case report identifies the objectives, methods, and results of a feasibility pilot conducted at Partners Healthcare from May to September, 2006. A candidate set of results were identified, approved for release, programmed into Patient Gateway, Partners' secure patient portal, and studied. Patient and practice feedback was positive. No noticeable rise in patient concerns was observed by practice staff or through patient surveys. One-half of patients who viewed results accessed reference information linked to a result. Organizational and practice-level issues necessary to support continued rollout are described. PMID- 17911671 TI - An evolving systems-based methodology for healthcare planning. AB - Healthcare planning seems beset with problems at all hierarchical levels. These are caused by the 'soft' nature of many of the issues present in healthcare planning and the high levels of complexity inherent in healthcare services. There has, in recent years, been a move to utilize systems thinking ideas in an effort to gain a better understanding of the forces at work within the healthcare environment and these have had some success. This paper argues that systems-based methodologies can be further enhanced by metrication and modeling which assist in exploring the changed emergent behavior of a system resulting from management intervention. The paper describes the Holon Framework as an evolving systems based approach that has been used to help clients understand complex systems (in the education domain) that would have application in the analysis of healthcare problems. PMID- 17911672 TI - Model-centric approaches for the development of health information systems. AB - Modeling is used increasingly in healthcare to increase shared knowledge, to improve the processes, and to document the requirements of the solutions related to health information systems (HIS). There are numerous modeling approaches which aim to support these aims, but a careful assessment of their strengths, weaknesses and deficiencies is needed. In this paper, we compare three model centric approaches in the context of HIS development: the Model-Driven Architecture, Business Process Modeling with BPMN and BPEL and the HL7 Development Framework. The comparison reveals that all these approaches are viable candidates for the development of HIS. However, they have distinct strengths and abstraction levels, they require local and project-specific adaptation and offer varying levels of automation. In addition, illustration of the solutions to the end users must be improved. PMID- 17911673 TI - Software engineering principles applied to large healthcare information systems- a case report. AB - Sao Paulo is the largest city in Brazil and one of the largest cities in the world. In 2004, Sao Paulo City Department of Health decided to implement a Healthcare Information System to support managing healthcare services and provide an ambulatory health record. The resulting information system is one of the largest public healthcare information systems ever built, with more than 2 million lines of code. Although statistics shows that most software projects fail, and the risks for the Sao Paulo initiative were enormous, the information system was completed on-time and on-budget. In this paper, we discuss the software engineering principles adopted that allowed to accomplish that project's goals, hoping that sharing the experience of this project will help other healthcare information systems initiatives to succeed. PMID- 17911674 TI - Towards modeling and simulation of integrated social and health care services for elderly. AB - In order to estimate the impact of an innovation on a segment of the health care system under certain assumptions such as different possible regulatory or financing schemes (scenarios) prior to its diffusion, one must understand the dynamic behavior of the entire system with its essential control loops. Aim of this feasibility study was to explore the potential of System Dynamics (SD) modeling for this purpose. First, a UML-based modeling of an Innovative Care for Elderly (ICE) system for provision of integrated social and health care services to elderly living at home was done. Then monetary and quality of life aspects of the social and health care system were described by two coarse SD models. On these models the impact of the introduction of the ICE system under certain assumption (scenarios) was studied, based on data from the German Health Expenditure and German Federal Statistics Office. The simulations show plausible behavior, however, are not yet detailed enough for a final conclusion. A major problem is missing data for setting model parameters: estimates had to be made. In conclusion, SD modeling might be a useful method for studying impacts of the diffusion of an innovation in the health for elderly sector, but more research is needed. PMID- 17911675 TI - Healthcare delivery systems: designing quality into health information systems. AB - To ensure that quality is 'engineered in' a holistic, integrated and quality approach is required, and Total Quality Management (TQM) principles are the obvious foundations for this. This paper describes a novel approach to viewing the operations of a healthcare provider where electronic means could be used to distribute information (including electronic fund settlements), building around the Full Service Provider core. Specifically, an approach called the "triple pair flow" model is used to provide a view of healthcare delivery that is integrated, yet detailed, and that combines the strategic enterprise view with a business process view. PMID- 17911676 TI - A mobile data collection tool for workflow analysis. AB - Faulty exchange and impaired access to clinical information is a major contributing factor to the incidence of medical error and occurrence of adverse events. Traditional methods utilized for systems analysis and information technology design fail to capture the nature of information use in highly dynamic healthcare environments. This paper describes a study designed to identify information task components in a cardiovascular intensive care unit and the development of an observational data collection tool to characterize the use of information in this environment. Direct observation can be a time-consuming process and without easy to use, reliable and valid methods of documentation, may not be reproducible across observers or settings. The following attributes were found to be necessary components for the characterization of information tasks in this setting: purpose, action, role, target, mode, and duration. The identified information task components were incorporated into the design of an electronic data collection tool to allow coding of information tasks. The reliability and validity of this tool in practice is discussed and an illustration of observational data output is provided. PMID- 17911677 TI - Towards sustainable e-health networks: does modeling support efficient management and operation? AB - e-Health Networks require cost-effective approaches for routine operation to achieve long-lasting sustainability. By using a model to represent (i) the network's enterprise functions, (ii) the applications used and (iii) the physical implementations, the tasks of management, adapting to changes and providing continued maintenance can be effectively supported. The paper discusses approaches for modeling, assesses their usefulness for the above tasks and decides on the use of the 3LGM meta model. Based on this concept, three ways for modeling the specific properties of an e-Health network are presented, leading to the decision to represent the hospitals involved in only one layer. As a result the model derived is presented, assessed and proved to support strategic management, day-to-day maintenance and documentation. PMID- 17911679 TI - A conceptual model of computerised hospital information system (CHIS) use in South Africa. AB - The aim of this project is to contribute to limiting the risk of CHIS (computerised hospital information system) failure by identifying factors which are associated with the successful implementation of CHISs in district and regional hospitals in South Africa (SA). Case studies were conducted in four regional hospitals in the Western Cape province of SA to obtain data about user perceptions of the success or lack of success of the CHISs in use. A conceptual model of CHIS use has been developed based on the results of the case studies, in order to assist in the interpretation of the differing experiences across the hospitals. Key factors in the conceptual model are perception of usefulness of the CHIS and management commitment to ensuring CHIS success, which in turn are related to effective use of CHIS and/or CHIS outputs, and allocation of resources for CHIS further development. Further development of the model will be influenced by the next phase of this project: a survey of district and regional hospitals in two SA provinces. PMID- 17911678 TI - The openEHR Java reference implementation project. AB - The openEHR foundation has developed an innovative design for interoperable and future-proof Electronic Health Record (EHR) systems based on a dual model approach with a stable reference information model complemented by archetypes for specific clinical purposes.A team from Sweden has implemented all the stable specifications in the Java programming language and donated the source code to the openEHR foundation. It was adopted as the openEHR Java Reference Implementation in March 2005 and released under open source licenses. This encourages early EHR implementation projects around the world and a number of groups have already started to use this code. The early Java implementation experience has also led to the publication of the openEHR Java Implementation Technology Specification. A number of design changes to the specifications and important minor corrections have been directly initiated by the implementation project over the last two years. The Java Implementation has been important for the validation and improvement of the openEHR design specifications and provides building blocks for future EHR systems. PMID- 17911680 TI - Are problem-oriented medial records (POMR) suitable for use in GPs' daily practice? AB - Problem-oriented functions have been implemented in almost all Belgian GPs' software systems since 2003. We therefore investigated whether some of them - especially the explicit linking procedure between treatments or referrals and the relevant problems - can be used by GPs in their current daily practice. In 2005, within the Belgian ResoPrim project, we organized data collection, mainly around the theme of "hypertension and cardiovascular risk factors", by 26 volunteer GPs' practices using three different software systems. Data were collected prospectively over six weeks in early 2005, and retrospectively for 2004. In this paper we report only on the part of the study that aimed to assess the linking procedure. For all patients and hypertensive patients alike, the key indicators used were the percentage of (problem-) linked drugs among the drugs extracted, the percentage of anti-hypertensive (problem-) linked drugs among anti hypertensive drugs extracted, and the percentage of (problem-) linked referrals among the number of referrals extracted. For all patients, the data collected relate to 10,914 contacts (7,831 patients) in 2005, and to 74,878 contacts (16,813 patients) in 2004. Large variations were observed per software system and GP, and also over time. The percentage of linked drugs rose from 2% (2004, two GPs) to 36% (2005, fourteen GPs). For linked referrals the percentage was 65% in 2004 vs. 75% in 2005. Our study shows that some functions related to the problem oriented patient record were spontaneously used by GPs in daily practice. This use increased during collaboration with the primary care research network. This increase was not restricted to the theme of data collection (i.e. not restricted to hypertensive patients, to anti-hypertensive drugs or to links with cardiovascular problems). PMID- 17911681 TI - Can an EPR support the concept of family-centred, individualized developmental care of premature infants and newborns? AB - At the University Children's Hospital Heidelberg the concept of 'Developmental, Family-Centred, Individual Care of Premature Infants and Newborns' was introduced to support optimal growth of premature infants. This interdisciplinary concept requires cooperation of different specialists. A well operating communication is a precondition for such cooperation. As a patient's record is not only used for storing information but also for exchanging information, the question was if a complete electronic patient record (EPR), in contrast to the existing patient's record, could sensibly support this new concept of care. To answer this question the whole communication of the staff in the infants ward was analysed using different observation methods. These observations delivered several issues which showed that an EPR could improve communication and workflow. Therefore an EPR for the neonatology at the University Children's Hospital Heidelberg can now be designed on the basis of our communication concept. PMID- 17911682 TI - Multiple detection modalities and disease natural history of breast cancer. AB - Multiple detection modalities have increasingly gained attention in population based screening. However, the disease natural history and its efficacy have been barely addressed. We reviewed a series of articles addressing multiple detection modalities including mammography, ultrasound and magnetic resonance image between 1995 and 2005. A stochastic model was developed to estimate transition parameters pertaining to the disease natural history defined by multiple detection modalities. The effectiveness of the combination of ultrasound or magnetic resonance image (MRI) with mammography was projected using a series of computer simulation models. The results indicated that multiple detection modalities may lead to reduced mortality. However, the benefit and the selection of detection modalities are affected by biological factors including age, breast tissue type and histological type. In addition, other social factors may also affect the utilization of multiple detection modalities. PMID- 17911683 TI - Challenges in telemedicine and eHealth: lessons learned from 20 years with telemedicine in Tromso. AB - The Norwegian Centre for Telemedicine (NST) has, over the past two decades, contributed to the development and implementation of telemedicine and ehealth services in Norway. From 2002, NST has been a WHO Collaboration Center for telemedicine. In August 1996, Norway became the first country to implement an official telemedicine fee schedule making telemedicine services reimbursable by the national health insurer. Telemedicine is widely used in Northern Norway. Since the late 1980's, the University Hospital of North-Norway has experience in the following areas: teleradiology, telepathology, teledermatology, teleotorhinolaryngology (remote endoscopy), remote gastroscopy, tele echocardiography, remote transmission of ECGs, telepsychiatry, teleophthalmology, teledialysis, teleemergency medicine, teleoncology, telecare, telegeriatric, teledentistry, maritime telemedicine, referrals and discharge letters, electronic delivery of laboratory results and distant teaching for healthcare personnel and patients. Based on the result achieved, the health authority in North-Norway plans to implement several large-scale telemedicine services: Teleradiology (incl. solutions for neurosurgery, orthopedic, different kinds of surgery, nuclear medicine, acute traumatic and oncology), digital communication and integration of patient data, and distant education. In addition, the following services will also be considered for large-scale implementation: teledialysis, prehospital thrombolysis, telepsychiatry, teledermatology. Last in line for implementation are: pediatric, district medical center (DMS), teleophthalmology and ear-nose-throat (ENT). PMID- 17911684 TI - A satellite infrastructure for health early warning in post-disaster health management. AB - The risk of epidemics and emerging or re-emerging diseases such as avian flu, tuberculosis, malaria and other vector-borne diseases, is rising. These risks can be contained with prevention, early warning, and prompt management. Despite progress in information technology, communication is still a bottleneck for health early warning and response systems in post-disaster situations. This paper presents Satellites for Epidemiology (SAFE), a component-based interoperable architecture for health early warning that employs satellite, radio, and wireless networks, geographic information systems, integration technology, and data mining to promptly identify and respond to a disease outbreak. In a post-disaster situation, a mobile health emergency coordination center is established and integrated to public health services for health monitoring. The added-value of SAFE for post-disaster health management will be demonstrated as part of an earthquake readiness exercise regarding a typhoid fever epidemic, in the island of Crete. Advanced communication and data mining techniques in SAFE offer new tools to the "Epidemic Intelligence" and contribute to advanced preparedness and prompt response by lifting communication barriers, promoting collaboration, and reducing the isolation of affected areas. PMID- 17911685 TI - Remote support for stroke rehabilitation: MyHeart's "neurological rehabilitation" concept. AB - Stroke is a serious neurological accident which accounts for a wide fraction of the healthcare costs in industrialized societies. Recovery from stroke and other neurological accidents usually include motor rehabilitation, maintained for several months, and logopedic training for the recovery of cognitive and speech abilities. The MyHeart consortium is addressing several aspects of cardiovascular diseases' management by combining clothes with embedded biomedical sensors and information technologies. One of the application areas is especially devoted to supporting Neurological Rehabilitation (NR). This article describes how MyHeart's Concept NR is structured and how technologies are leveraged to support both motor rehabilitation and speech/cognitive training. Information technology and garment embedded sensors, combined, permit assisted training both at the clinic and at home, after discharge from the intensive care unit. PMID- 17911686 TI - Effects of a computerized cardiac teletriage decision support system on nurse performance: results of a controlled human factors experiment using a mid fidelity prototype. AB - A gap exists in cardiac care between known best practices and the actual level of care administered. To help bridge this gap, a proof of concept interface for a PDA-based decision support system (DSS) was designed for cardiac care nurses engaged in teletriage. This interface was developed through a user-centered design process. Quality of assessment, quality of recommendations, and number of questions asked were measured. Cardiac floor nurses' assessment quality performance, but not their recommendation quality performance, improved with the DSS. Nurses asked more questions with the DSS than without it, and these additional questions were predominantly classifiable as essential or beneficial to a good assessment. The average participant satisfaction score with the DSS was above neutral. PMID- 17911687 TI - Opportunities and barriers for mobile health in New Zealand. AB - Ownership and the use of mobile technologies greatly exceed those of personal desktop computer systems and countries throughout the world are beginning to understand how these technologies can enhance the delivery of healthcare (m health). This paper reviews the opportunities and barriers for m-health and describes a study to understand its potential in New Zealand. A survey consisting of a questionnaire and in-depth interviews was used to reveal clinician and service provider attitudes to m-health. The general perception is that m-health will be an increasing component of future healthcare with many opportunities for empowering patients, delivering convenience care, and supporting carers as well as offering the potential for more effective public health and lifestyle broadcasting. Participants recognised several barriers to the acceptance and sustainability of m-health, identifying privacy of information and device form factor as major concerns. PMID- 17911688 TI - Applying mobile and pervasive computer technology to enhance coordination of work in a surgical ward. AB - Collaboration, coordination, and communication are crucial in maintaining an efficient and smooth flow of work in an operating ward. This coordination, however, often comes at a high price in terms of unsuccessfully trying to get hold of people, disturbing telephone calls, looking for people, and unnecessary stress. To accommodate this situation and to increase the quality of work in operating wards, we have designed a set of pervasive computer systems which supports what we call context-mediated communication and awareness. These systems use large interactive displays, video streaming from key locations, tracking systems, and mobile devices to support social awareness and different types of communication modalities relevant to the current context. In this paper we report qualitative data from a one-year deployment of the system in a local hospital. Overall, this study shows that 75% of the participants strongly agreed that these systems had made their work easier. PMID- 17911689 TI - Feasibility and usability of a home monitoring concept based on mobile phones and near field communication (NFC) technology. AB - Utilization of mobile information and communication technologies in home monitoring applications is becoming more and more common. The mobile phone, acting as a patient terminal for patients suffering from chronic diseases, provides an active link to the caregiver to transmit health status information and receive feedback. In such a concept the usability is still limited by the necessity of entering the values via the mobile phone's small keypad. The near field communication technology (NFC), a touch-based wireless interface that became available recently, may improve the usability level of such applications significantly. The focus of this paper is to describe the development of a prototype application based on this technology embedded in a home monitoring system. The feasibility and usability of this approach are evaluated and compared with concepts used in previous approaches. The high quantifier with respect to overall usability indicates that NFC may be the technology of choice for some tasks in home monitoring applications. PMID- 17911690 TI - Using personal digital assistants and patient care algorithms to improve access to cardiac care best practices. AB - In order to facilitate knowledge transfer between specialists and generalists and between experts and novices, and to promote interdisciplinary communication, there is a need to provide methods and tools for doing so. This interdisciplinary research team developed and evaluated a decision support tool (DST) on a personal digital assistant (PDA) for cardiac tele-triage/tele-consultation when the presenting problem was chest pain. The combined human factors methods of cognitive work analysis during the requirements-gathering phase and ecological interface design during the design phase were used to develop the DST. A pilot clinical trial was conducted at a quaternary cardiac care hospital over a 3-month period. During this time, the DST was used by the nine nursing coordinators who provide tele-triage/tele-consultation 24/7. This clinical trial validated the design and demonstrated its usefulness to advanced cardiac care nurses, its potential for use by nurses less experienced in cardiac care, and for its potential use in an interdisciplinary team environment. PMID- 17911691 TI - Improving systems interoperability with model-driven software development for healthcare. AB - An aging population and an increase in chronically ill patients demand teamwork treatment models. To support these with information systems, interoperability is a prerequisite. Model-driven software development (MDSD) with special healthcare extensions can enable reuse of components and improve conformance to international standards. In this paper, a MDSD HealthCare Framework is proposed and demonstrated for homecare services. Using the framework, information systems will improve their conformance to international standards and the interoperability with other systems. PMID- 17911692 TI - Conformance testing of interoperability in health information systems in Finland. AB - Conformance testing has been studied extensively but the current state of the art varies very much in different software companies in the healthcare domain. In this paper we present findings of a survey of conformance testing of interface specifications in the healthcare domain. The survey was conducted among software companies and their customers, i.e. hospital districts, in Finland. The findings of the survey show that the quality of the applications should be improved, that there is a reluctance to use external interface testing services, and that software vendors do not adequately fulfill customers' requirements in relation to testing. However, conformance testing is considered to be very important and should be improved. The main contribution of this study is the analysis of the findings and we give some recommendations for healthcare organizations, software companies, and authorities to improve the conformance testing related to the interoperability of the applications. PMID- 17911693 TI - Connecting public health and clinical information systems by using a standardized methodology. AB - To meet the challenge for efficient, high quality and sustainable care, health systems in developed and increasingly in developing countries require extended communication and cooperation between all principals involved in citizen's care. The challenge also concerns supporting information systems, which demand interoperation with public health, bioinformatics, genomics, administrative, governmental, and other sources of data. The paper describes an architecture development methodology for modeling the integration between clinical and public health information systems that harmonizes existent standardized modeling approaches and integrates HL7 domain knowledge. An integration-architecture for information sharing between public health surveillance and clinical information systems is derived demonstrating the feasibility of the proposed methodology. Predominantly, a harmonized process for analysis, design, implementation and maintenance of semantically interoperable information systems based on formal grammars is discussed in some detail. PMID- 17911694 TI - Clinical and translational science sustainability: overcoming integration issues between electronic health records (EHR) and clinical research data management systems "separate but equal". AB - The use of health information technology (HIT) is growing rapidly for patient care systems required to test, diagnose and treat patients, and to bill for these services. Today's Electronic Health Record (EHR) systems are a response to this pressure, enabling feature rich computer-assisted decisions and communication. And even though EHR benefits dramatically outweigh the costs, required investments are nonetheless significant. Continuing to invest in HIT at a revolutionary rate is unsustainable given institutional financial constraints and continuing reimbursement cuts. Future improvements must come from new treatments, test methods, drugs and devices - from research. But data management information systems for clinical research receive less funding than patient care systems, and in less coherent ways. It is easy to imagine using the high cost, patient-based EHRs for clinical research data management, and thus accelerate the speed of translating new medical discoveries into standard practice. But taking this step requires thoughtful planning to overcome significant technology, legal/regulatory, policy, process, and administrative issues. PMID- 17911695 TI - The FA4CT algorithm: a new model and tool for consumers to assess and filter health information on the Internet. AB - BACKGROUND: eHealth-literate consumers, consumers able to navigate and filter credible information on the Internet, are an important cornerstone of sustainable health systems in the 21(st) century. Various checklists and tools for consumers to assess the quality of health information on the Internet have been proposed, but most fail to take into account the unique properties of a networked digital environment. METHOD: A new educational model and tool for assessing information on the Internet has been designed and pilot tested with consumers. The new proposed model replaces the "traditional" static questionnaire/checklist/ rating approach with a dynamic, process-oriented approach, which emphasizes three steps consumers should follow when navigating the Internet. FA4CT (or FACCCCT) is an acronym for these three steps: 1) Find Answers and Compare [information from different sources], 2) Check Credibility [of sources, if conflicting information is provided], 3) Check Trustworthiness (Reputation) [of sources, if conflicting information is provided]. In contrast to existing tools, the unit of evaluation is a "fact" (i.e. a health claim), rather than a webpage or website. RESULTS: Formative evaluations and user testing suggest that the FA4CT model is a reliable, valid, and usable approach for consumers. CONCLUSION: The algorithm can be taught and used in educational interventions ("Internet schools" for consumers), but can also be a foundation for more sophisticated tools or portals, which automate the evaluation according to the FA4CT algorithm. PMID- 17911696 TI - WRAPIN: a tool for patient empowerment within EHR. AB - Legal and technologic trends are making medical records progressively more patient-accessible. In the near future, information technology may make it even easier to provide patients a chance to review their records. One may wonder, however, about the practical use of this technology by patients. Understanding his/her own health record will certainly be one of the main concerns of patients. WRAPIN has been designed to provide patients and citizens with trusted health information. It will help to determine the reliability of documents by checking the ideas contained against established benchmarks, and enable users to determine the relevance of a given document from a page of search results. First, we present what is, in our opinion, the most original and important patient-centred WRAPIN characteristics and functionalities. Then, we compare these characteristics with those of representatives of two main trends in information retrieval: systems based on the popularity of web sites, and on the clustering of web sites. This comparison shows that, even though patients are tempted to use popular search engines, these are not sufficiently specialized in the medical domain to help them understand their own HER.. Finally, we discuss the complexity of medical readings over the Internet and the efforts that are still required in this domain. PMID- 17911697 TI - How do clinicians search for and access biomedical literature to answer clinical questions? AB - This paper presents a retrospective data analysis on how 75 clinicians searched for and accessed biomedical literature from an online information retrieval system to answer six clinical scenarios. Using likelihood ratio measures to quantify the impact of documents on a decision, and a graphical representation to model clinicians' journeys of accessing documents, this analysis reveals that clinicians did not necessarily arrive at the same answer after having accessed the same document, and that documents did not influence clinicians in the same manner. A possible explanation for these phenomena is that people experience cognitive biases during information searching which influence their decision outcome. This analysis raises the hypotheses that people experience the anchoring effect, order effects, exposure effect and reinforcement effect while searching for information and that these biases may subsequently influence the way decisions are made. PMID- 17911698 TI - Blogs, wikis, and discussion forums: attributes and implications for clinical information systems. AB - Informaticians increasingly view clinical information systems as asynchronous communication systems instead of data processing tools. Outside of health care, popular web technologies like blogs, wikis, and discussion forums have proven to be platforms for effective asynchronous communication. These popular technologies have implications for improving the coordination of clinical care and social support. In order to appropriately evaluate these webbased tools for use in clinical information systems, it will be essential for the informatics community to formally identify the distinguishing attributes of these communication methodologies. The authors propose seven interpersonal and informational attributes to compare and contrast the purposes of blogs, wikis, and discussion forums. This attribute-based approach to analyzing emerging web technologies will lead to a better understanding of the design choices involved in web-based information systems. Two case studies demonstrate how informatics researchers and developers can consider these attributes in the design and evaluation of clinical information systems. PMID- 17911700 TI - A mobile phone based remote patient monitoring system for chronic disease management. AB - Rising concern over the poor state of chronic disease management led to the user informed design and development of a home tele-monitoring system. Focus groups with patients and primary care providers guided the research team towards a design that would accommodate the workflow and concerns of the healthcare providers and the low use and comfort with technology found among the patient population. The system was trialed in a before-and-after pilot study of 34 patients with diabetes and hypertension. Findings demonstrate a significant improvement in systolic and diastolic blood pressure. An RCT beginning in 2007 is being conducted to confirm these findings. It is hypothesized that this user centred approach, utilizing focus groups, iterative design and human factors methods of evaluation, will lead to the next-generation of home tele-monitoring applications that are more intuitive, less cumbersome, and ultimately bring about greater patient compliance and better physician management. PMID- 17911699 TI - From intermediation to disintermediation and apomediation: new models for consumers to access and assess the credibility of health information in the age of Web2.0. AB - This theoretical paper discusses the model that, as a result of the social process of disintermediation enabled by digital media, traditional intermediaries are replaced by what this author calls apomediaries, which are tools and peers standing by to guide consumers to trustworthy information, or adding credibility to information. For apomediation to be an attractive and successful model for consumers, the recipient has to reach a certain degree of maturity and autonomy. Different degrees of autonomy may explain differences in information seeking and credibility appraisal behaviours. It is hypothesized that in an apomediated environment, tools, influential peers and opinion leaders are the primary conveyors of trust and credibility. In this environment, apomediary credibility may become equally or more important than source credibility or even message credibility. It is suggested to use tools of network analysis to study the dynamics of apomediary credibility in a networked digital world. There are practical implications of the apomediation model for developers of consumer health websites which aspire to come across as "credible: Consumers need and want to be able to be co-creators of content, not merely be an audience who is broadcasted to. Web2.0 technology enables such sites. Engaging and credible Web sites are about building community and communities are built upon personal and social needs. PMID- 17911701 TI - How to enhance integrated care towards the personal health paradigm? AB - For improving quality and efficiency of health delivery under the well-known burdens, the health service paradigm has to change from organization-centered over process-controlled to personal health. The growing complexity of highly distributed and fully integrated healthcare settings can only be managed through an advanced architectural approach, which has to include all dimensions of personal health. Here, ICT, medicine, biomedical engineering, bioinformatics and genomics, legal and administrative aspects, terminology and ontology have to be mentioned. The Generic Component Model allows for different domains' concept representation and aggregation. Framework, requirements, methodology and process design possibilities for such a future-proof and meanwhile practically demonstrated approach are discussed in detail. The deployment of the Generic Component Model and the concept representation to biomedical engineering aspects of eHealth are touched upon as essential issues. PMID- 17911702 TI - Developing information technology attitude scales for health (ITASH). AB - This paper reports on the ongoing development and testing of a set of scales designed to elicit the attitudes of United Kingdom healthcare staff towards Information Technology. The scales were developed in the light of existing literature. Testing included a test-retest completion by over 100 staff from various disciplines in three National Health Service Trusts and a comparison with an existing scale. Exploratory principal components factor analysis identified three components, with a loading of > 3.1. This pattern of item grouping could be understood and interpreted as representing 'efficiency of care', 'education, training and development' and 'control'. The three scales comprise the Information Technology Attitude Scales for Health (ITASH). The results suggest that the developed scales together comprise a measure that can be used to establish staff attitudes towards IT use in the healthcare sector. Those undertaking research in this area might consider employing the scales to measure the factors that influence attitudes towards IT. Additionally, employers might usefully draw on the scales as they aim to support staff in IT use and embed IT systems within the healthcare workplace. PMID- 17911703 TI - Development of patient centric virtual organizations (PCVOs) in clinical environment for patient information management. AB - A novel Virtual Organization framework which incorporates wireless technology support is presented in the research work. The Virtual Organization is designed for a clinical environment to provide better patient information management and enhanced collaborative working of multidisciplinary care teams. The analysis studies the current clinical practices and looks at the general patient information resource structure currently in use for patient care. Based on this problem analysis and current requirements of the multi-disciplinary care team members, we propose a generic and sustainable Patient Centric Virtual Organization (PCVO) framework to complement the functionality of the existing infrastructure by incorporating wireless technologies support for improved patient information provision at the point of care. The preliminary results of the study identify and classify the specific point of care tasks suited to appropriate information resources needed by the care team members. This paper concentrates on the patient information management aspects brought in by incorporating wireless technologies at the point of care using patient information resources in a decentralized and distributed computing environment. This applied research is carried out in the secondary and tertiary care sector in the cancer domain. For the analysis and results of the pilot project, we have used a case study of a local NHS Cancer Hospital. PMID- 17911704 TI - SPIRS: a framework for content-based image retrieval from large biomedical databases. AB - With the increasing use of medical images in clinical medicine, disease research, and education, the need for methods that effectively archive, query, and retrieve these images by their content is underscored. This paper presents the implementation of a Web-based retrieval system called SPIRS (Spine Pathology & Image Retrieval System) at the U. S. National Library of Medicine that demonstrates recent developments in shape representation and retrieval from a large dataset of 17,000 digitized x-ray images of the spine and associated text records. Users can search these images by providing a sketch of the vertebral outline or selecting an example vertebral image and some relevant text parameters. Pertinent pathology on the image/sketch can be annotated and weighted to indicate importance. This hybrid text-image query yields images containing similar vertebrae along with relevant fields from associated text records, which allows users to examine the pathologies of vertebral abnormalities. Initial experiments with SPIRS have demonstrated the potential for this system, particularly on a large dataset of clinical images. PMID- 17911705 TI - The quality of reporting of health informatics evaluation studies: a pilot study. AB - The quality of reporting of evaluations in Health Informatics is considered to be open for improvement. STARE-HI provides guidance for improved reporting. This study aims at establishing the quality of reporting prior to dissemination of STARE-HI. All evaluation papers in three leading Medical Informatics Journals published in 2005 have been assessed with the aid of STARE-HI. A total of 48 papers have been analysed with respect to title, abstract, keywords and study context. Main results indicate that the type of study is seldom reported in the title, that the structured abstract contains more information than an unstructured abstract, that important keywords are often forgotten. Also important information about the system assessed is often not mentioned explicitly. Our conclusion is that there is room for improvement in the reporting of evaluations in health informatics. STARE-HI can be helpful in suggesting a structure for evaluation papers in Health Informatics and issues that should be included in such papers. PMID- 17911706 TI - Exploring the unintended consequences of computerized physician order entry. AB - This paper summarizes the foci, activities, methods, and results of a three-year research project. Using a mixed methods approach, the Physician Order Entry Team has identified and categorized the unintended consequences of computerized physician order entry (CPOE). After analyzing 380 examples of unintended adverse consequences, the team described in detail nine major types and conducted a national survey in the U.S. to discover how hospitals recognize and deal with unintended consequences. With the assistance of a panel of experts, the team identified strategies for managing unintended adverse consequences and outlined contents of a toolkit for CPOE implementers for addressing them. PMID- 17911707 TI - Impact of health care information technology on hospital productivity growth: a survey in 17 acute university hospitals. AB - The quantification of the added value of information technologies (IT) in the health sector is a major issue for decision-makers and health care professionals. This paper relates the application of an economic production function in hospitals with different integration levels of their clinical information systems (CIS). The study concerns 17 university hospitals within the Assistance Publique Hopitaux de Paris group that were followed from 1998 to 2005. Using an extended Cobb-Douglas production function, yearly incomes (Y) were correlated with three inputs: capital (K), labor (L) and IT expenses (T). The results indicate that incomes are significantly and positively associated with the three input variables with elasticity coefficients: alpha, beta and gamma of 0.81, 0.17, and 0.09 that appear to be in the range of values found in secondary and tertiary sectors. The IT elasticity coefficient (gamma) is higher in the subgroup of 6 hospitals that integrate, or started to integrate, a complete CIS within the study period than in the 11 reference hospitals. In a general production function, hospital costs appear to be positively connected to the level of IT expenses, capital and labor. Calculations in two subgroups of AP-HP hospitals divided according to the importance of the IT integration level seem to indicate that the more the clinical information system is integrated, the more its influence is positive in hospital production. The results of this first survey are sufficiently encouraging to try to refine them (better granularity) and to spread them in time (over a longer period) and space (to other hospital structures). PMID- 17911708 TI - Risk management and measuring productivity with POAS--point of act system. AB - The concept of our system is not only to manage material flows, but also to provide an integrated management resource, a means of correcting errors in medical treatment, and applications to EBM through the data mining of medical records. Prior to the development of this system, electronic processing systems in hospitals did a poor job of accurately grasping medical practice and medical material flows. With POAS (Point of Act System), hospital managers can solve the so-called, "man, money, material, and information" issues inherent in the costs of healthcare. The POAS system synchronizes with each department system, from finance and accounting, to pharmacy, to imaging, and allows information exchange. We can manage Man, Material, Money and Information completely by this system. Our analysis has shown that this system has a remarkable investment effect - saving over four million dollars per year - through cost savings in logistics and business process efficiencies. In addition, the quality of care has been improved dramatically while error rates have been reduced - nearly to zero in some cases. PMID- 17911709 TI - Connecting the dots: creation of an electronic regional infection control network. AB - The prevalence of Methicillin-resistant Staphylococcus aureus (MRSA) and Vancomycin-resistant Enterococci (VRE) continues to increase dramatically worldwide. Successful programs to reduce infection rates of resistant organisms require regional or national compliance with strict infection control measures and feedback on implementation and reduced rates. We partnered with local infection control professionals (ICPs) and leveraged our existing electronic network to create a comprehensive city-wide network to track and uniformly respond to patients admitted with a history of MRSA or VRE. We successfully standardized and included electronic data from four out of six of the major healthcare systems within Indianapolis. We created tailored abstracts to deliver key infection control data to ICPs when a MRSA patient is admitted to a participating hospital. We created web-based data entry forms for ICPs to modify and enter new infection control data. This paper describes our design and initial implementation of a working electronic regional infection control network. PMID- 17911710 TI - Dealing with ecological fallacy in preparations for influenza pandemics: use of a flexible environment for adaptation of simulations to household structures in local contexts. AB - Development of strategies for mitigating the severity of a new influenza pandemic is a global public health priority. The aim of this study is to examine effects on simulation outcomes caused by variations in local socio-demographic data. We used a spatially explicit geo-physical model of a virtual city as a baseline and employed an ontology-modeling tool to construct alternative population distributions and household structures. We found that adjustment for the case when single parents in practice were cohabiting led to a higher reproduction rate than that observed for a population with the highest formally recorded share of households with >2 children. When antivirals sufficient to protect 10 percent of the population were administered to schoolchildren, a preliminary effect on the reproduction rate was observed. This effect was eliminated when the household structure was adjusted for cohabiting single parents. Nations have been encouraged to develop estimates of morbidity and mortality during a possible pandemic outbreak. In order to deal with ecological fallacy, the present results suggest that this recommendation can be extended also to local communities. PMID- 17911711 TI - Effectiveness of a chronic disease surveillance systems for blood pressure monitoring. AB - Information Technology (IT) enables health care providers to manage patients with chronic conditions through identification, follow up and administration of specific interventions. In our setting, we developed a surveillance system for chronic diseases. The aim of this study was to show its efficacy on monitoring blood pressure throughout a cluster randomized controlled trial. Patients without blood pressure registries (condition 1) or with high blood pressure measurements (condition 2) were randomized to be detected by the surveillance system or to receive usual care. The proportion of patients with at least one blood pressure measurement within three months of follow up was 49.9% (207 patients) in the intervention group and 37% (195) in the control group (p<0.001) for condition 1. And 61% (224) vs. 50% (239) respectively (p=0.002) for condition 2. Patients under the surveillance system have higher proportion of blood pressure measurements, showing this study an improvement on the process of care with this IT tool. PMID- 17911712 TI - Nurses and computers. An international perspective on nurses' requirements. AB - This paper reports the findings from a Florence Nightingale Foundation Travel Scholarship undertaken by the author in the spring of 2006. The aim of the visit was to explore nurses' attitudes towards, and experiences of, using computers in their practice, and the requirements that they have to encourage, promote and support them in using ICT. Nurses were found to be using computers mainly for carrying out administrative tasks, such as updating records, rather than as information tools to support evidence based practice, or patient information needs. Nurses discussed the systems they used, the equipment provided, and their skills, or more often their lack of skills. The need for support was a frequent comment, most nurses feeling that it was essential that help was available at the point of need, and that it was provided by someone, preferably a nurse, who understood the work context. Three groups of nurses were identified. Engagers; Worried Willing and Resisters. The report concludes that pre-registration education has a responsibility to seek to ensure that newly qualified nurses enter practice as engagers. PMID- 17911713 TI - The need for global certification in the field of health informatics: some ethical issues. AB - In the past, the training of health information professionals (HIPs) has focussed almost exclusively on technical matters, the concerns of software developers and purveyors have essentially centred on security and functionality, and health care providers have mainly worried about costs and efficiency. This paper outlines some ethical threats that are ignored by such a purely technical focus and argues that because of the increasing globalization of health care delivery through e Health, and because of the international threats to confidentiality posed by legislation such as the US Patriot Act, the health informatics community should pursue a project of global certification for HIPs that includes information ethics as an integral component. It also argues that a corresponding certification process for health care institutions and software developers should be initiated. PMID- 17911714 TI - Using fieldwork in analyzing ethical issues related to IT in health care. AB - This paper describes how an understanding of everyday conflicts that have ethical implications - what we call 'situated ethics'- can be explored through ethnographic field techniques in healthcare settings. Our approach to ethics is followed by findings from two ethnographic case studies focussing on issues arising as information technologies such as electronic patient records and automatic drug dispensing machines are introduced into varied health sector workplaces. By close and careful observation of these technologies in use and by incorporating narrative accounts from different perspectives the complexity and entangledness of real life occurrences are revealed. Our data suggest that several types of ethical issues (e.g., issues related to intellectual property, literacy, standardization, transparency, work ethics, and equitable allocation of resources) can be identified through fieldwork, and can have an impact on identification of everyday ethics in healthcare. PMID- 17911715 TI - Handling consent to patient data access in a hospital setting. AB - The right to use patient data in treatment is based on the conditions of a need to know and patient consent. In electronic health records, these two conditions can be applied in various ways. We study the handling of consent in two Norwegian hospitals, with a view to how access control and consent handling can be integrated across electronic systems that process patient data. A workshop was held, where two consent handling scenarios were simulated, one in-hospital, the other external. Activities were identified and tied to roles and to the documents and systems used. Electronic systems were found to support the execution of the scenarios to some extent. The electronic functions used in-hospital were consent storage and logging of access; access control was not sufficient. When sharing information externally, the typical approach is a declaration signed by the patient or a referral; such external information sharing should be supported by specific functionality. A first step towards integrated access control is integrated consent handling. PMID- 17911716 TI - A day in the life of a clinical research coordinator: observations from community practice settings. AB - One of the goals of the NIH Roadmap Initiative is to re-engineer the national clinical research enterprise, with an emphasis on information technology solutions. Understanding end-users' workflow is critical to developing technology systems that are grounded in the context of the users' environment and are designed to fulfill their needs. Community practices are becoming the prevailing setting for conducting clinical research. Few studies have assessed clinical research workflow in such settings. We have conducted a series of investigations to model the workflow and have previously reported on some basic aspects of it, like the lack of information systems to support the workflow. In this paper we describe finer details of the workflow, using results of observational studies. These findings highlight the needs and inefficiencies that suggest the kind of information system that must be developed to enhance collaboration, communication and improve efficiency. This preliminary investigation also opens ground for more extensive studies to further elucidate the workflow. PMID- 17911717 TI - A survey of U.S.A. acute care hospitals' computer-based provider order entry system infusion levels. AB - We developed and fielded a survey to help clinical information system designers, developers, and implementers better understand the infusion level, or the extent and sophistication of CPOE feature availability and use by clinicians within acute care hospitals across the United States of America. In the 176 responding hospitals, we found that CPOE had been in place a median of 5 years and that the median percentage of orders entered electronically was 90.5%. Greater than 96% of the sites used CPOE to enter pharmacy, laboratory and imaging orders; 82% were able to access all aspects of the clinical information system with a single sign on; 86% of the respondents had order sets, drug-drug interaction warnings, and pop-up alerts even though nearly all hospitals were community hospitals with commercial systems; and 90% had a CPOE committee with a clinician representative in place. While CPOE has not been widely adopted after over 30 years of experimentation, there is still much that can be learned from this relatively small number of highly infused (with CPOE and clinical decision support) organizations. PMID- 17911718 TI - e-Healthcare in India: critical success factors for sustainable health systems. AB - As healthcare enterprises seek to move towards an integrated, sustainable healthcare delivery model an IT-enabled or e-Healthcare strategy is being increasingly adopted. In this study we identified the critical success factors influencing the effectiveness of an e-Healthcare strategy in India. The performance assessment criteria used to measure effectiveness were increasing reach and reducing cost of healthcare delivery. A survey of healthcare providers was conducted. Analytic Hierarchy Process (AHP) and Interpretive Structural Modeling (ISM) were the analytical tools used to determine the relative importance of the critical success factors in influencing effectiveness of e Healthcare and their interplay with each other. To succeed in e-Healthcare initiatives the critical success factors that need to be in place are appropriate government policies, literacy levels, and telecommunications and power infrastructure in the country. The focus should not be on the IT tools and biomedical engineering technologies as is most often the case. Instead the nontechnology factors such as healthcare provider and consumer mindsets should be addressed to increase acceptance of, and enhance the effectiveness of, sustainable e-Healthcare services. PMID- 17911719 TI - Health information systems adoption: findings from a systematic review. AB - Earlier evaluation studies on Health Information Systems (HIS) adoption have highlighted a large number of adoption problems that were attributed to the lack of fit between technology, human and organisation factors. Lessons can be learned from these evaluation studies by identifying the most important factors of HIS adoption. In order to study the adoption issue, a qualitative systematic review has been performed using a recently introduced framework, known as HOT-fit (Human, Organisation and Technology fit). The paper identifies and highlights the following critical adoption factors: technology (ease of use, system usefulness, system flexibility, time efficiency, information accessibility and relevancy); human (user training, user perception, user roles, user skills, clarity of system purpose, user involvement); organisation (leadership and support, clinical process, user involvement, internal communication, inter organisational system, as well as the fit between them. The findings can be used to guide future system development and inform relevant decision making. PMID- 17911720 TI - Telemedicine portal and standardized clinical documents. AB - The paper describes the experience of creating a flexible telemedicine portal intended to improve a business process of information exchanges needed to manage real-time telemedicine events. The portal structure allows use of digitally signed standardized clinical documents based on the HL7 Clinical Document Architecture. PMID- 17911721 TI - The development of an information system and installation of an Internet web database for the purposes of the occupational health and safety management system. AB - This paper is based on the research on the possible structure of an information system for the purposes of occupational health and safety management. We initiated a questionnaire in order to find the possible interest on the part of potential users in the subject of occupational health and safety. The depiction of the potential interest is vital both for the software analysis cycle and development according to previous models. The evaluation of the results tends to create pilot applications among different enterprises. Documentation and process improvements ascertained quality of services, operational support, occupational health and safety advice are the basics of the above applications. Communication and codified information among intersted parts is the other target of the survey regarding health issues. Computer networks can offer such services. The network will consist of certain nodes responsible to inform executives on Occupational Health and Safety. A web database has been installed for inserting and searching documents. The submission of files to a server and the answers to questionnaires through the web help the experts to perform their activities. Based on the requirements of enterprises we have constructed a web file server. We submit files so that users can retrieve the files which they need. The access is limited to authorized users. Digital watermarks authenticate and protect digital objects. PMID- 17911722 TI - Reliability assessment of home health care services. AB - In this paper, a model of reliability assessment of services in Home Health Care Delivery is presented. Reliability is an important quality dimension for services and is included in non-functional requirements of a system. A stochastic Markov model for reliability assessment is applied to patient communication services, in the field of home health care delivery. The methodology includes the specification of scenarios, the definition of failures in scenarios as well as the application of the analytical model. The results of the methodology reveal the critical states of the Home Health Care System and recommendations for improvement of the services are proposed. The model gives valuable results in predicting service reliability and, independently of the error types, it can be applied to all fields of Regional Health Network (RHN). PMID- 17911723 TI - From the description of activities to the identification of risks for clinical management: a proposal of building, merging and sharing knowledge representations of care processes. AB - Management of clinical processes and hospital activities takes advantage of business process reengineering methodology. It is now recognized that care process modeling must integrate the definition of goals and the assessment of risk. Two kinds of issues have been outlined: 1) the lack of an integrated model to identify and describe processes and their components according to a functional point of view; and 2) an increasing amount of documents that hospital staff members have to create, collect, index and maintain. As initial models focused only on a structural view of activities, we reviewed different sources of standards and norms to extract and classify a set of metadata aimed at describing any activity and its outcomes. The model includes links to structured terminologies to name attributes or value them. An object-oriented information model has been created and implemented to test the relevance and the feasibility of the modeling approach. Conceptually speaking, this model gives opportunity to bridge tacit and explicit knowledge. Practically speaking, limits to generalization remain partly due to the lack of a template processes database. PMID- 17911724 TI - Near-miss and hazard reporting: promoting mindfulness in patient safety education. AB - Patient safety efforts advocate for transforming healthcare from a culture of blame to a non-punitive "culture of safety." One of the most challenging hurdles is to encourage healthcare practitioners to be mindful about their activities. To promote mindfulness during clinical education for entry-level nurses, we developed a web-based dangerous situation and near miss reporting system for a cohort of baccalaureate nursing students (N=156). For this curricular innovation project, we provided wireless handheld devices to students who were required to submit one report for each of their clinical days in the medical-surgical, pediatric, psychiatric, obstetrics and community settings. During a ten-week period, students submitted 1487 reports. Of these, 63% were dangerous situations and 37% were near misses. The most frequently occurring dangerous situations were poor infection control practices. The most commonly reported near misses were medication errors. Free text comments from students identified inadequate patient identification and poor documentation as commonly occurring dangerous situations observed. PMID- 17911725 TI - A territory-wide electronic health record--from concept to practicality: the Hong Kong experience. AB - In Hong Kong, a pilot project is being undertaken to implement a web-based electronic patient record system to allow integrated, real time patient based information to be shared in clinics, private and public hospitals. Such sharing aims to ensure that complete and accurate healthcare information is available to citizens' multiple points of care through a stable IT system. A challenge is to share this electronic information whilst ensuring privacy and security. Hong Kong has achieved its initial goals and pioneered in building a territory-wide electronic health record (EHR). This paper will outline the tasks involved, approach, method used and initial review of the pilot project. Barriers to implementation are discussed and critical success factors are identified. PMID- 17911726 TI - Northern Territory HealthConnect: shared electronic health record service implementation experiences and benefits realised in indigenous health. AB - This presentation summarises the learnings from the HealthConnect Northern Territory (HCNT) Shared Electronic Health Record Service (SEHR) from Trial to Implementation and the emerging benefits realized as the project is implemented across the Northern Territory of Australia. The presentation: * explores the challenges and experiences of implementing a SEHR service in urban and in some of the remotest regions on the Australian continent. * demonstrates the emerging health benefits e-Health can provide in enabling the sharing of medical information between public and private health service providers in particular the service delivery and benefits provided to a highly mobile Indigenous population who currently experience the greatest health problems and experience difficulties accessing continuum of care created by factors which include remoteness, mobility and communication. * explores the evolvement of the "opt in" consumer consent model adopted by the Territory for the implementation of the HCNT SEHR.* advises of plans for future development, which inform other implementations, and NeHTA standards development for the implementation of the National SEHR Service. * Informs project plans to incorporate NeHTA standards as they are developed and transition the HCNT SEHR to the National SEHR Service when implemented recognising the importance of developing strong partnerships with key stakeholders, in particular consumers, health care providers and system vendors who inform project development and implementation. PMID- 17911727 TI - A simulation-based performance analysis of a National Electronic Health Record System. AB - This paper addresses through simulation experiments a number of technical issues which are raised during the development and operation of a National Electronic Health Record System (NEHRS). The simulation experiments represent the NEHRS performance for a variety of technological infrastructures, within the context of a realistic scenario. The scenario includes the estimation of the delays created in queues during the exchange of Electronic Patient Records (EPR) between different health service points. It is essential to clarify the delays derive from LAN and Internet technologies, the EPR encryption/decryption, the HL7 message generation/parsing, and the databases. The results of this study identify how a number of technical aspects influence the NEHRS development and operation. PMID- 17911728 TI - Principles-based medical informatics for success--how Hong Kong built one of the world's largest integrated longitudinal electronic patient records. AB - Since 1994, the Hospital Authority has been developing and deploying clinical applications at its constituent 41 hospitals and 121 clinics. The Clinical Management System (CMS) is now used by over 4000 doctors and 20000 other clinicians on a daily basis to order, document and review care. The territory wide Electronic Patient Record (ePR) has given clinicians an integrated, longitudinal, lifelong view of a patient's record. Today the CMS and ePR form an essential clinical and management tool to the Hospital Authority. The CMS handles two million clinical transactions per day, and the ePR has over 6TB of data covering 57 million episodes for 7.9 million patients. This paper describes how the HA has taken a principles-based approach to Medical Informatics to achieve its success in the enterprise-wide deployment and deep utilization of a comprehensive clinical information system. PMID- 17911729 TI - Knowledge-level querying of temporal patterns in clinical research systems. AB - Managing time-stamped data is essential to clinical research activities and often requires the use of considerable domain knowledge. Adequately representing this domain knowledge is difficult in relational database systems. As a result, there is a need for principled methods to overcome the disconnect between the database representation of time-oriented research data and corresponding knowledge of domain-relevant concepts. In this paper, we present a set of methodologies for undertaking knowledge level querying of temporal patterns, and discuss its application to the verification of temporal constraints in clinical-trial applications. Our approach allows knowledge generated from query results to be tied to the data and, if necessary, used for further inference. We show how the Semantic Web ontology and rule languages, OWL and SWRL, respectively, can support the temporal knowledge model needed to integrate low-level representations of relational data with high-level domain concepts used in research data management. We present a scalable bridge-based software architecture that uses this knowledge model to enable dynamic querying of time-oriented research data. PMID- 17911731 TI - Confidentiality preserving audits of electronic medical record access. AB - Failure to supply a care provider with timely access to a patient's medical record can lead to patient harm or death. As such, healthcare organizations often endow care providers with broad access privileges to electronic medical record (EMR) systems. In doing so, however, care providers may access a patient's record without legitimate purpose and violate patient privacy. Healthcare privacy officials use EMR access logs to investigate potential violations. The typical log is limited in its information, so that it is often necessary to merge access logs with other information systems. The problem with this practice is that sensitive information about patients and care providers may be disclosed in the process. In this paper, we present a privacy preserving technique that enables linkage of disparate health information systems without revealing sensitive information. The technique permits any number of vested parties to contribute to audit investigations without learning information about those being investigated. We motivate the protocol in a real world medical center and then generalize the protocol for implementation in existing healthcare environments. PMID- 17911730 TI - A discussion about the importance of laws and policies for data sharing for public health in the People's Republic of China. AB - This paper introduces the current status of data sharing in the People's Republic of China. It discusses barriers to data sharing and proposes three key solutions to overcome these barriers in China. The establishment of national laws and policies for data sharing is considered the key prerequisite to ensuring the successful implementation of resource sharing activities in public health. Driven by established laws and policies, the relevant operational models should be developed. It is also important to have strategies in place to ensure the established laws and policies are implemented by various organizations in different jurisdictions. These discussions are supported by relevant local and international evidence. PMID- 17911732 TI - Design of a decentralized reusable research database architecture to support data acquisition in large research projects. AB - The diagnostic and therapeutic processes, as well as the development of new treatments, are hindered by the fragmentation of information which underlies them. In a multi-institutional research study database, the clinical information system (CIS) contains the primary data input. An important part of the money of large scale clinical studies is often paid for data creation and maintenance. The objective of this work is to design a decentralized, scalable, reusable database architecture with lower maintenance costs for managing and integrating distributed heterogeneous data required as basis for a large-scale research project. Technical and legal aspects are taken into account based on various use case scenarios. The architecture contains 4 layers: data storage and access are decentralized at their production source, a connector as a proxy between the CIS and the external world, an information mediator as a data access point and the client side. The proposed design will be implemented inside six clinical centers participating in the @neurIST project as part of a larger system on data integration and reuse for aneurism treatment. PMID- 17911733 TI - The Cancer Biomedical Informatics Grid (caBIG): infrastructure and applications for a worldwide research community. AB - Information explosion and new advances in high throughput experiments have challenged biomedical research, and suggested a future in which inter institutional and international collaborations will be the norm. The cancer Biomedical Informatics Grid is an ambitious initiative launched by the US National Cancer Institute to develop a network of tools, data, and researchers to support translational and clinical research in oncology, with an ultimate goal to improve cancer care for patients. The three year pilot phase of caBIG ends in 2007, and has engaged over 900 clinicians, scientists, and patient advocates as developers, adopters, and workspace participants. Progress has been demonstrated in creating tools and building prototype grid architecture for collaborative research. Accomplishments in the pilot phase set the stage for extension of the community into other biomedical domains and for federation of the caBIG enterprise with similar initiatives in other scientific areas and in other countries. PMID- 17911734 TI - The integration of grid resources into a portal for research collaboratories. AB - Information technology (IT) infrastructures for research collaboratories and virtual research businesses are essential elements of science in medicine. The sustainability of the infrastructure depends on how it becomes a normal element of the scientific process, including its financing as part of a scientist's workbench that can be easily configured to their needs. The results should also be distributed through a suitable platform to other research networks. The development of such an infrastructure serves all parties - mostly the funding agencies. This objective could be achieved by the integration of several portals of different research networks and the interfacing between these portals to maximize the usability and minimize the cost. PMID- 17911735 TI - Biomedical data mining in clinical routine: expanding the impact of hospital information systems. AB - In this paper we want to describe how the promising technology of biomedical data mining can improve the use of hospital information systems: a large set of unstructured, narrative clinical data from a dermatological university hospital like discharge letters or other dermatological reports were processed through a morpho-semantic text retrieval engine ("MorphoSaurus") and integrated with other clinical data using a web-based interface and brought into daily clinical routine. The user evaluation showed a very high user acceptance - this system seems to meet the clinicians' requirements for a vertical data mining in the electronic patient records. What emerges is the need for integration of biomedical data mining into hospital information systems for clinical, scientific, educational and economic reasons. PMID- 17911736 TI - The Swedish National Pharmacy Register. AB - To achieve a safer future prescribing, the Swedish government has introduced a mandatory registration of all drugs dispensed at pharmacies. The medication history in the register may be accessed online by registered individuals, prescribers and pharmacists. After 15 months of action, the prevalence of individuals with dispensed drugs in the Swedish population was 71.0% (6,424,487/9,047,752); women 78.8% and men 63.1%. The incidence rate for individuals with dispensed drugs was estimated as 12.4 (1,000*111,960/9,047,752) per month and 1, 000 inhabitants. The mean number of dispensed prescriptions was 12.1 (median 6, Q1-Q3 2-15) per individual. For the elderly (age group 80-89), the mean number of dispensed prescriptions was 27.8 during the study period (median 24,Q1-Q3 13-38); women 28.8 and men 26.1. When introducing a National Pharmacy Register, containing personal drug information for the majority of the population, issues on security, confidentiality and ethics have to be taken into consideration. The lack of widespread secure digital signatures in health care may delay general availability. To clinically evaluate individual medication history, the relatively high prevalence of dispensed drugs in the population, seems to justify the National Pharmacy Register. PMID- 17911737 TI - The use of existing low-cost technologies to enhance the medical record documentation using a summary patient record [SPR]. AB - The Institute of Medicine has described the Electronic Medical Record [EMR] as an essential technology for health care that improves patient safety and the quality of care when compared to traditional paper-based records. [1,2,3] Despite major financial expenditures on health information technology related to EMRs in developed countries such as North America, Britain and Australia, success rates for successful implementations have been low. One specific domain of information management relates to the communication of health care between those involved in the care process. Summarization of patient histories is a core component of EMR systems and assists in the communication of health care. This paper demonstrates how the utilization of simple technologies, can lead to the successful implementation of the Summary Patient Record [SPR] component of an EMR system. PMID- 17911738 TI - A new approach in nursing documentation: community nursing case. AB - This paper presents the development of electronic documentation for community nursing using a system approach. Documentation is viewed as an information model for organizing and managing processes. The community nurse plans the nursing process after gathering and evaluating information on the patient's health and his/her family status. Documentation is thus considered to be a basis for the successful work of the health team and as a way of ensuring quality in nursing. The paper describes a prototype software model for e-documentation in community nursing together with its evaluation in practice. PMID- 17911739 TI - A survey of the effects of the full computerized nursing records system on sharing nursing records among health professionals. AB - In the last decade computerized nursing records systems (CNRSs) have been implemented at many hospitals around the world. Several effects of the CNRS were expected; the improvement of the quality of medicine and nursing care, the increased efficiency and the reduction of the cost. This study focused on the effects of the CNRS on access and sharing of nursing records among various health professionals. Timely access and availability of nursing records should improve the quality of medicine and nursing care. In 2003, we conducted a survey of the effectiveness of the CNRS on access and availability of nursing records among health professionals. We found that the CNRS contributed to the multidisciplinary sharing of nursing records without increasing the overall time spent on nursing documentation at most hospitals. However, effective sharing of nursing records even among nurses through the use of the CNRS did not occur at many nursing divisions. PMID- 17911740 TI - New method of realization of nursing diagnosis based on 3N in an electronic medical record system. AB - An electronic medical recording (EMR) system is enlightened as a solution for deciding which nursing diagnosis was selected on the basis of a computerized system; it can help collect and analyze lots of diverse data in an objective way. But there are few reports of successful electronic nursing diagnosis on EMR systems. This study was to develop the objective decision system prior to nursing diagnosis and to adopt it in the Severance EMR system. We adopt a new concept, situational variables, as the key elements of the nursing process, based on the items of nursing intervention. It enables appropriate nursing diagnosis through complex clinical objective data (laboratory results, vital signs, etc.). Through these elements between nursing intervention and nursing diagnosis, we can create reliable evidence-based nursing diagnoses successfully, and make possible fast settlement of nursing intervention in the various clinical fields. PMID- 17911741 TI - Integration of longitudinal electronic records in a large healthcare enterprise: the U.S. Veterans Health Administration experience. AB - The U.S. Veterans Health Administration (VHA) provides care to some 5.2 million patients spread across the continental United States, Alaska, Hawaii, Puerto Rico, and the Philippines. Sites of care include 157 medical centers, nearly 900 outpatient clinics, long-term facilities, and home care. Over the last 10 years, major changes in the nature of VHA healthcare have imposed a requirement for longitudinal electronic health records and integration of those records across the enterprise at the point of care. VHA has now evolved through three generations of applications that support such integration. This paper reports on the VHA experience, points out lessons learned, and outlines future directions for electronic health record integration in VHA. PMID- 17911742 TI - The AMPATH medical record system: creating, implementing, and sustaining an electronic medical record system to support HIV/AIDS care in western Kenya. AB - Providing high-quality HIV/AIDS care requires high-quality, accessible data on individual patients and visits. These data can also drive strategic decision making by health systems, national programs, and funding agencies. One major obstacle to HIV/AIDS care in developing countries is lack of electronic medical record systems (EMRs) to collect, manage, and report clinical data. In 2001, we implemented a simple primary care EMR at a rural health centre in western Kenya. This EMR evolved into a comprehensive, scalable system serving 19 urban and rural health centres. To date, the AMPATH Medical Record System contains 10 million observations from 400,000 visit records on 45,000 patients. Critical components include paper encounter forms for adults and children, technicians entering/managing data, and modules for patient registration, scheduling, encounters, clinical observations, setting user privileges, and a concept dictionary. Key outputs include patient summaries, care reminders, and reports for program management, operating ancillary services (e.g., tracing patients who fail to return for appointments), strategic planning (e.g., hiring health care providers and staff), reports to national AIDS programs and funding agencies, and research. PMID- 17911743 TI - Sao Paulo city health information system--a case report. AB - Sao Paulo is the largest city in Brazil and one of the largest in the world. In 2004, Sao Paulo City Department of Health decided to implement a Healthcare Information System that would support managing healthcare services and provide an ambulatory health record. The system was designed to build on from national standards that identify healthcare workers, organizations and the relationships among them and reuse as much as possible existing concepts and software. At the same time, the system should reduce fragmentation, not only by integrating existing redundant or competing systems but also by providing a framework in which new modules would be naturally integrated to each other. Today, the web based system, known as SIGA Saude, runs in more than 370 healthcare units, processes some 8 thousand appointment scheduling daily and more than 10 thousand high-cost procedure authorizations monthly. This paper profiles the Sao Paulo City and its needs and describes the project, obstacles and results so far. PMID- 17911744 TI - Experience in implementing the OpenMRS medical record system to support HIV treatment in Rwanda. AB - The challenge of scaling up HIV treatment in Africa has led to a new emphasis on improving health systems in impoverished areas. One aspect of this is the development and deployment of electronic medical record systems to support HIV and TB treatment. In this paper we describe the design and implementation of a new medical record architecture to support an HIV treatment program in rural Rwanda. The architecture is called OpenMRS and it has been developed to address the problem of configuring EMR systems to suit new sites, languages and diseases. OpenMRS uses a data dictionary called the concept dictionary to represent all the possible data items that can be collected. This allows new items to be added to the system by non-programmers. In addition, there are form creation tools that use drag and drop web technologies to simplify form construction. The OpenMRS system was first implemented in Kenya in February 2006 and then in Rwanda in August 2006. The system is now functioning well and we are developing extensions to improve the support for the clinic. These include improved, easy to use reporting tools, support for additional clinical problems including nutrition and child health, better database synchronization tools, and modules to collect laboratory data and support the pharmacy. The system is also in use in South Africa and Lesotho and is being deployed in Tanzania and Uganda. PMID- 17911745 TI - Multilingual information retrieval in thoracic radiology: feasibility study. AB - Most essential information contained in the electronic medical record is stored as text, and this imposes several difficulties on automated data extraction and retrieval. Natural language processing is an approach that can unlock clinical information from free texts. The proposed methodology uses the specialized natural language processor MEDLEE developed for the English language. To use this processor on Portuguese medical texts, chest X-ray reports were machine translated (MT) into English. The result of serial coupling of MT and NLP is tagged text that needs further investigation for extracting clinical findings. This experiment's objective was to investigate normal reports and reports with device description on a set of 165 chest X-ray reports. We obtained sensitivity and specificity of 1 and 0.71 for the first condition, and 0.97 and 0.97 for the second. The reference was formed by the opinions of two radiologists. The results of this experiment indicate the viability of extracting clinical findings from chest X-ray reports through coupling MT and NLP. PMID- 17911746 TI - Large-scale evaluation of a medical cross-language information retrieval system. AB - We propose an approach to multilingual medical document retrieval in which complex word forms are segmented according to medically relevant morpho-semantic criteria. At its core lies a multilingual dictionary, in which entries are equivalence classes of subwords, i.e. semantically minimal units. Using two different standard test collections for the medical domain, we evaluate our approach for six languages covered by our system. PMID- 17911747 TI - EHR query language (EQL)--a query language for archetype-based health records. AB - OpenEHR specifications have been developed to standardise the representation of an international electronic health record (EHR). The language used for querying EHR data is not as yet part of the specification. To fill in this gap, Ocean Informatics has developed a query language currently known as EHR Query Language (EQL), a declarative language supporting queries on EHR data. EQL is neutral to EHR systems, programming languages and system environments and depends only on the openEHR archetype model and semantics. Thus, in principle, EQL can be used in any archetype-based computational context. In the EHR context described here, particular queries mention concepts from the openEHR EHR Reference Model (RM). EQL can be used as a common query language for disparate archetype-based applications. The use of a common RM, archetypes, and a companion query language, such as EQL, semantic interoperability of EHR information is much closer. This paper introduces the EQL syntax and provides example clinical queries to illustrate the syntax. Finally, current implementations and future directions are outlined. PMID- 17911748 TI - Design and evaluation of a temporal, graph-based language for querying collections of patient histories. AB - Giving clinicians and researchers the ability to easily retrieve and explore relevant fragments of patient histories would greatly facilitate quality assurance, patient followup and research on patient treatment processes. Established database query languages are inconvenient for such exploration, and may also be too complex for users with limited backgrounds in informatics. We believe that understandability can be increased in return for a sacrifice of some of the power of expression found in general query languages. In order to design a specialized query language, we have collected and synthesized a tentative list of requirements. Based on these requirements, we have designed and implemented Practice Explorer, a prototype for visual query of collections of patient histories, and evaluated the understandability of its query language by testing with medical students. The results indicate that parts of the language are intuitive enough for users to understand without demonstrations, examples, feedback or assistance. They also provide some lessons for future work in this area. PMID- 17911749 TI - Evaluation of a simple method for the automatic assignment of MeSH descriptors to health resources in a French online catalogue. AB - BACKGROUND: The growing number of resources to be indexed in the catalogue of online health resources in French (CISMeF) calls for curating strategies involving automatic indexing tools while maintaining the catalogue's high indexing quality standards. OBJECTIVE: To develop a simple automatic tool that retrieves MeSH descriptors from documents titles. METHODS: In parallel to research on advanced indexing methods, a bag-of-words tool was developed for timely inclusion in CISMeF's maintenance system. An evaluation was carried out on a corpus of 99 documents. The indexing sets retrieved by the automatic tool were compared to manual indexing based on the title and on the full text of resources. RESULTS: 58% of the major main headings were retrieved by the bag-of-words algorithm and the precision on main heading retrieval was 69%. CONCLUSION: Bag-of words indexing has effectively been used on selected resources to be included in CISMeF since August 2006. Meanwhile, on going work aims at improving the current version of the tool. PMID- 17911750 TI - Template-based data entry for general description in medical records and data transfer to data warehouse for analysis. AB - General descriptions in medical records are so diverse that they are usually entered as free text into an electronic medical record, and the resulting data analysis is often difficult. We developed and implemented a template-based data entry module and data analyzing system for general descriptions. We developed a template with tree structure, whose content master and entered patient's data are simultaneously expressed by XML. The entered structured data is converted to narrative form for easy reading. This module was implemented in the EMR system, and is used in 35 hospitals as of October, 2006. So far, 3725 templates (3242 concepts) have been produced. The data in XML and narrative text data are stored in the EMR database. The XML data are retrieved, and then patient's data are extracted, to be stored in the data ware-house (DWH). We developed a search assisting system that enables users to find objective data from the DWH without requiring complicated SQL. By using this method, general descriptions in medical records can be structured and made available for clinical research. PMID- 17911751 TI - Challenges and methodology for indexing the computerized patient record. AB - Patient records contain most crucial documents for managing the treatments and healthcare of patients in the hospital. Retrieving information from these records in an easy, quick and safe way helps care providers to save time and find important facts about their patient's health. This paper presents the scalability issues induced by the indexing and the retrieval of the information contained in the patient records. For this study, EasyIR, an information retrieval tool performing full text queries and retrieving the related documents has been used. An evaluation of the performance reveals that the indexing process suffers from overhead consequence of the particular structure of the patient records. Most IR tools are designed to manage very large numbers of documents in a single index whereas in our hypothesis, one index per record, which usually implies few documents, has been imposed. As the number of modifications and creations of patient records are significant in a day, using a specialized and efficient indexation tool is required. PMID- 17911752 TI - Experiments with hierarchical concept-based search. AB - Many digital libraries use hierarchical indexing schema, such as MeSH to enable concept based search in the retrieval phase. However, improving or outperforming the traditional full text search isn't trivial. We present an extensive set of experiments using a hierarchical concept based search retrieval method, applied in addition to several baselines, within the Vaidruya search and retrieval framework. Concept Based Search applied in addition to a low baseline is outperforming significantly, especially when queried on concepts in the third level and using disjunction within the hierarchical trees. PMID- 17911753 TI - StructConsult: structured real-time wet read consultation infrastructure to support patient care. AB - Our research addresses how to improve physician to physician communication of patient information, and how to prevent lapses of patient care as they are referred to other clinicians within the healthcare system. The wet read consultation is defined as a rapid response to a clinical question posed by a referring physician to a clinical specialist. This research involves the development of an imaging-based wet read consultation system called StructConsult (SC), which facilitates communication between non-imaging specialist (i.e., primary care physician (PCP), emergency room (ER) physician, or referring physician), and an imaging specialist-radiologist. To facilitate data mining and effective recall, SC utilizes a data model based on the Digital Image Communications in Medicine (DICOM) standard for grayscale presentation state and structured reporting. SC requires information from four sources: (a) patient specific demographics, clinical hypothesis, and reason for exam, (b) sentinel image capture from a DICOM image study, (c) direct capture of radiologist's image operations and annotations, and (d) radiologist's response to the chief compliant, and the reason for examination. SC allows users to add additional functionality to a Picture Archiving System to improve patient care. PMID- 17911754 TI - Refining DICOM for pathology--progress from the IHE and DICOM pathology working groups. AB - For making medical decisions, healthcare professionals require that all necessary information is both correct and easily available. We address the issue of integrating anatomical pathology department information into the electronic healthcare enterprise. The pathology workflow from order to report, including specimen processing and image acquisition was modelled. An integration profile - pathology general workflow- was created in the framework of the Integrating the Healthcare Enterprise (IHE). This Integration Profile relies on 8 transactions based on HL7 or DICOM standards. An important issue was to define information entities (order, imaging study and report) and real-world objects (specimen, tissue sample, slide, etc.). Joint efforts between IHE and DICOM WG26 has resulted in a proposed common model for "specimen" usable for both HL7 and DICOM transactions related to anatomic pathology. PMID- 17911755 TI - A generic, web-based clinical information system architecture using HL7 CDA: successful implementation in dermatological routine care. AB - The requirements of highly specialized clinical domains are often underrepresented in hospital information systems (HIS). Common consequences are that documentation remains to be paper-based or external systems with insufficient HIS integration are used. This paper presents a solution to overcome this deficiency in the form of a generic framework based on the HL7 Clinical Document Architecture. The central architectural idea is the definition of customized forms using a schema-controlled XML language. These flexible form definitions drive the user interface, the data storage, and standardized data exchange. A successful proof-of-concept application in a dermatologic outpatient wound care department has been implemented, and is well accepted by the clinicians. Our work with HL7 CDA revealed the need for further practical research in the health information standards realm. PMID- 17911756 TI - Analyzing the key variables in the adoption process of HL7. AB - The intention of this paper is to provide an analysis of variables that could affect the adoption of HL7 message standard. Based on the review of 33 cases where HL7 was successfully implemented the authors present relevant evidence related to inherent limitation of HL7. The result from this study indicates that it is necessary to enhance the standard to overcome particular limitations and facilitate the implementation of inter-institutional software interfaces based on HL7. PMID- 17911757 TI - An XML model of an enhanced data dictionary to facilitate the exchange of pre existing clinical research data in international studies. AB - Pre-existing clinical research data sets exchanged in international epidemiology research often lack the elements needed to assess their suitability for use in multi-region meta-analyses or other clinical studies. While the missing information is generally known to local investigators, it is not contained in the files exchanged between sites. Instead, such content must be solicited by the study coordinating center though a series of lengthy phone and electronic communications: an informal process whose reproducibility and accuracy decays over time. This report describes a set of supplemental information needed to assess whether clinical research data from diverse research sites are truly comparable, and what metadata ("data about the data") should be preserved when a data set is archived for future use. We propose a structured Extensible Markup Language (XML) model that captures this information. The authors hope this model will be a first step towards preserving the metadata associated with clinical research data sets, thereby improving the quality of international data exchange, data archiving, and merged-data research using data collected in many different countries, languages and care settings. PMID- 17911758 TI - Framework for clinical data standardization based on archetypes. AB - Standardization of data is a prerequisite to achieve semantic interoperability in any domain. This is even more important in the healthcare sector where the need for exchanging health related data among professional and institutions is not an exception but the rule. Currently, there are several international organizations working on the definition of electronic health record architectures, some of them based on a dual-model approach. We present both an archetype modeling framework and LinkEHR-ED, an archetype editor and mapping tool for transforming existing electronic healthcare data which do not conform to a particular electronic healthcare record architecture into compliant electronic health records extracts. In particular, archetypes in LinkEHR-ED are formal representations of clinical concepts built on a particular reference model but enriched with mapping information to data sources which define how to extract and transform existing data in order to generate standardized XML documents. PMID- 17911759 TI - Reaching standards for dissemination: a case study. AB - At what stage are web applications ready for dissemination? A set of standards for efficacy, effectiveness and readiness for dissemination of (prevention) interventions has been developed by the Society for Prevention Research (SPR) (Flay et al., 2005) [1]. In this case study paper, we examine these standards criteria with reference to MoodGYM, an automated web application designed to reduce depression symptoms using cognitive behaviour therapy training. We examine evidence for its efficacy, its effectiveness in real world situations, and its capacity to meet recommended standards for dissemination. We conclude that MoodGYM has substantially met the standards of evidence required for dissemination. This successful application of the SPR standards to the evaluation of a web application, suggests that these prevention standards might be usefully applied to web/Internet context. Web applications might be assessed for dissemination by the International Society for Research into Internet Interventions (ISRII), or some other professional organization to assist policy makers in making decisions about the funding, adoption and promotion of applications. PMID- 17911760 TI - A discrete time-space geography for epidemiology: from mixing groups to pockets of local order in pandemic simulations. AB - The World Health Organization urges all nations to develop and maintain national influenza preparedness plans. Important components of such plans are forecasts of morbidity and mortality based on local social and geographic conditions. Most methodologies for simulations of epidemic outbreaks are implicitly based on the assumption that the frequency and duration of social contacts that lead to disease transmission is affected by geography, i.e. the spatial distribution of physical meeting places. In order to increase the effectiveness of the present methods for simulation of infectious disease outbreaks, the aim of this study is to examine two social geographic issues related to such models. We display how the social geographic characteristics of mixing networks, in particular when these significantly deviate from the random-mixing norm, can be represented in order to enhance the understanding and prediction of epidemic patterns in light of a possible future destructive influenza pandemic. We conclude that social geography, social networks and simulation models of directly transmitted infectious diseases are fundamentally linked. PMID- 17911761 TI - A national study of eHealth standardization in Finland--goals and recommendations. AB - The role of standards is constantly increasing in health services, electronic health records, and eHealth applications. There are many areas of standardization which affect the healthcare work and health information systems. On a national level, the organization of the development and support for standardization should be a key priority. This paper summarizes a national study in Finland which reviewed the current status of eHealth standardization and made recommendations for the national standardization and the use of standards. The recommendations are related to the organization model and balanced participation, international and cross-domain collaboration and quality assurance of standards-related activities. In addition, education and support services and support for introductions and pilots are recommended to improve the know-how of standards in system acquisitions. PMID- 17911762 TI - Building a womb-to-tomb health record in Hong Kong--an application of information architecture. AB - The Hospital Authority developed the Information Architecture (IA) model in 2002 to support a fast, robust, flexible and accurate electronic patient record (ePR) to meet the high-tempo health care environment in Hong Kong. With several successful applications in sharing data that were created for the same patients in various systems, the IA model was further developed to extend the longitudinal ePR to include one's fetal data as entered in the mother's record. This paper describes how various IA elements: Section, View, Form, Group, Entity, Content, Document supports the building of a true womb-to-tomb ePR for the HA patients. The future focus of Information Architecture in the HA will include building a Information Architecture Management System and linking the ePR with other patient records in the community. PMID- 17911763 TI - Another HISA--the new standard: health informatics--service architecture. AB - In addition to the meaning as Health Informatics Society of Australia, HISA is the acronym used for the new European Standard: Health Informatics - Service Architecture. This EN 12967 standard has been developed by CEN - the federation of 29 national standards bodies in Europe. This standard defines the essential elements of a Service Oriented Architecture and a methodology for localization particularly useful for large healthcare organizations. It is based on the Open Distributed Processing (ODP) framework from ISO 10746 and contains the following parts: Part 1: Enterprise viewpoint. Part 2: Information viewpoint. Part 3: Computational viewpoint. This standard is now also the starting point for the consideration for an International standard in ISO/TC 215. The basic principles with a set of health specific middleware services as a common platform for various applications for regional health information systems, or large integrated hospital information systems, are well established following a previous prestandard. Examples of large scale deployments in Sweden, Denmark and Italy are described. PMID- 17911764 TI - Protecting privacy while sharing medical data between regional healthcare entities. AB - Economies of scale, corporate partnerships and a need to increase the efficiency of Information Technology in the Healthcare sector are leading to the construction of Regional Health Information Organizations (RHIOs) across the United States. RHIOs are normally aligned by service provision given by particular healthcare payers (e.g. Blue Cross-Blue Shield, PacifiCare etc.) in particular geographies. Globalization has created a transient workforce that may require their healthcare provider access their patient data across several sovereign RHIOs. The barrier to enabling RHIO to RHIO collaboration lies in the need to respect the data disclosure policy of each RHIO, to adhere to the geography-specific healthcare legislation and also to not violate the express privacy wishes of the patient(s) involved. In this paper, we propose a data-level control called Sticky Policy Enforcement which allows sharing to occur across RHIOs, while adhering to the concerns mentioned. PMID- 17911765 TI - Pulling back the covers: technical lessons of a real-world health information exchange. AB - Several nations and local communities are striving to achieve widespread, secure exchange of clinical data between various health care providers and public health organizations. Most of the literature on health information exchange focuses on the financial, political, and privacy aspects of these initiatives. Perhaps just as important are the technical and organizational factors that have influenced development of data exchange methods and results. One mature network in the Midwestern United States has had success in establishing consistent, secure exchange of clinical data for more than ten years. Presented here are the technical lessons learned and design decisions made from this initiative with the hope that they can be used by others striving to connect disparate clinical information systems for the improvement of health care quality and safety. PMID- 17911766 TI - Geographically distributed complementary content-based image retrieval systems for biomedical image informatics. AB - There is a significant increase in the use of medical images in clinical medicine, disease research, and education. While the literature lists several successful systems for content-based image retrieval and image management methods, they have been unable to make significant inroads in routine medical informatics. This can be attributed to the following: (i) the challenging nature of medical images, (ii) need for specialized methods specific to each image type and detail, (iii) lack of advances in image indexing methods, and (iv) lack of a uniform data and resource exchange framework between complementary systems. Most systems tend to focus on varying degrees of the first two items, making them very versatile in a small sampling of the variety of medical images but unable to share their strengths. This paper proposes to overcome these shortcomings by defining a data and resource exchange framework using open standards and software to develop geographically distributed toolkits. As proof-of-concept, we describe the coupling of two complementary geographically separated systems: the IRMA system at Aachen University of Technology in Germany, and the SPIRS system at the U. S. National Library of Medicine in the United States of America. PMID- 17911767 TI - Utilizing SELinux to mandate ultra-secure access control of medical records. AB - Ongoing concerns have been raised over the effectiveness of information technology products and systems in maintaining privacy protection for sensitive data. The aim is to ensure that sensitive health information can be adequately protected yet still be accessible only to those that "need-to-know". To achieve this and ensure sustainability over the longer term, it is advocated that an alternative, stable and secure system architecture is required. This paper considers the adoption of a model targeted at health information that provides much higher degrees of protection. A purpose built demonstrator that was developed based on enterprise-level systems software products is detailed. The long term aim is to provide a viable solution by utilizing contemporary, commercially supported operating system and allied software. The advantages and limitations in its application with a medical database are discussed. The future needs in terms of research, software development and changes in organizational policy for healthcare providers, are outlined. PMID- 17911768 TI - Proposal of a French health identification number interoperable at the European level. AB - The French ministry of Health is setting up the Personal Medical Record (PMR). This innovative tool has long been expected by French Health Authorities, Associations of Patients, other Health's associations, those defending Individual Liberties and the French National Data Protection Authority. The PMR will lead to improvements in many areas such as Diagnosis (Research and monitoring) Healthcare (Management of emergencies, urgent situations, Temporal health monitoring and evaluation), Therapy (Cohorts of patients for Clinical trials and epidemiological studies). The PMR will foster safe healthcare management, clinical research and epidemiological studies. Nevertheless, it raises many important questions regarding duplicates and the quality, precision and coherence of the linkage with other health data coming from different sources. The currently planned identifying process raises many questions with regard to its ability to deal with potential duplicates and to perform data linkage with other health data sources. Through this article, using the electronic health records, we develop and propose an identification process to improve the French PMR. Our proposed unique patient identifier will guarantee the security, confidentiality and privacy of the personal data, and will prove to be particularly useful for health planning, health policies and research as well as clinical and epidemiological studies. Finally, it will certainly be interoperable with other European health information systems. We propose here an alternative identification procedure that would allow France to broaden the scope of its PMR project by making it possible to contribute to public health research and policy while increasing interoperability with European health information systems and preserving the confidentiality of the data. PMID- 17911769 TI - Clearinghouse: a teleradiology platform emphasizing security of data and communication. AB - The Clearinghouse application platform is a web based solution for secure digital exchange of radiological images and other clinical documents among authorized researchers and physicians. It implements a sophisticated security and role model to protect privacy and to minimize the risk of eavesdropping of patient data. The Clearinghouse serves as a centralized platform for distributed, distantly located medical research and health care. It is based on Open-Source software, thus ensuring continued support, maintenance, security and last but not least continuity of the platform. The use of the Clearinghouse minimizes turnaround times by superseding comparably slow and insecure conventional communication methods otherwise used for the exchange of radiological images and clinical documents, such as standard mail and courier services. Furthermore, it alleviates the integration of distantly located expert knowledge into diagnostic routines, culminating in an increased health care quality regardless of location of patients or physicians. PMID- 17911770 TI - K-Box: automatic structuring and exchange of medical documents based on the clinical documentation architecture (CDA). AB - Sustainable health systems should understand information as a resource which also has to be managed efficiently. Today's electronic documentation is only the mandatory first step to automatic information exchange between hospitals, physician offices, pharmacies and other participants. The K-Box is a modular system which allows different monolithic information systems to be connected and integrated by using the clinical document architecture (CDA) as the standard document exchange format. A prototype K-Box has been implemented and a trial run is scheduled. The workflow selected for the trial produces discharge summaries in CDA format. PMID- 17911771 TI - Using distributional analysis to semantically classify UMLS concepts. AB - The UMLS is a widely used and comprehensive knowledge source in the biomedical domain. It specifies biomedical concepts and their semantic categories, and therefore is valuable for Natural Language Processing (NLP) and other knowledge based systems. However, the UMLS semantic classification is not always accurate, which adversely affects performance of these systems. Therefore, it is desirable to automatically validate, or, when necessary, to semantically reclassify UMLS concepts. We applied a distributional similarity method based on syntactic dependencies and -skew divergence to classify concepts in the T033 Finding class in order to determine which ones were biologic functions or disorders. A gold standard of 100 randomly sampled concepts was created that was based on a majority annotation of three experts. Precision of 0.54 and recall of 0.654 was achieved by the top prediction; precision of 0.64 and recall of 0.769 was achieved by the top 2 predictions. Error analysis revealed problems in the current method, and provided insight into future improvements. PMID- 17911772 TI - A reappraisal of sentence and token splitting for life sciences documents. AB - Natural language processing of real-world documents requires several low-level tasks such as splitting a piece of text into its constituent sentences, and splitting each sentence into its constituent tokens to be performed by some preprocessor (prior to linguistic analysis). While this task is often considered as unsophisticated clerical work, in the life sciences domain it poses enormous problems due to complex naming conventions. In this paper, we first introduce an annotation framework for sentence and token splitting underlying a newly constructed sentence- and token-tagged biomedical text corpus. This corpus serves as a training environment and test bed for machine-learning based sentence and token splitters using Conditional Random Fields (CRFs). Our evaluation experiments reveal that CRFs with a rich feature set substantially increase sentence and token detection performance. PMID- 17911773 TI - Corpus-based error detection in a multilingual medical thesaurus. AB - Cross-language document retrieval systems require support by some kind of multilingual thesaurus for semantically indexing documents in different languages. The peculiarities of the medical sublanguage, together with the subjectivism of lexicographers' choices, complicates the thesaurus construction process. It furthermore requires a high degree of communication and interaction between the lexicographers involved. In order to detect errors, a systematic procedure is therefore necessary. We here describe a method which supports the maintenance of the multilingual medical subword repository of the MorphoSaurus system which assigns language-independent semantic identifiers to medical texts. Based on the assumption that the distribution of these semantic identifiers should be similar whenever comparing closely related texts in different languages, our approach identifies those semantic identifiers that vary most in distribution comparing language pairs. The revision of these identifiers and the lexical items related to them revealed multiple errors which were subsequently classified and fixed by the lexicographers. The overall quality improvement of the thesaurus was finally measured using the OHSUMED IR benchmark, resulting in a significant improvement of the retrieval quality for one of the languages tested. PMID- 17911774 TI - Defining medical words: transposing morphosemantic analysis from French to English. AB - Medical language, as many technical languages, is rich with morphologically complex words, many of which take their roots in Greek and Latin-in which case they are called neoclassical compounds. Morphosemantic analysis can help generate definitions of such words. This paper reports work on the adaptation of a morphosemantic analyzer dedicated to French (DeriF) to analyze English medical neoclassical compounds. It presents the principles of this transposition and its current performance. The analyzer was tested on a set of 1,299 compounds extracted from the WHO-ART terminology. 859 could be decomposed and defined, 675 of which successfully. An advantage of this process is that complex linguistic analyses designed for French could be successfully transferred to the analysis of English medical neoclassical compounds. Moreover, the resulting system can produce more complete analyses of English medical compounds than existing ones, including a hierarchical decomposition and semantic gloss of each word. PMID- 17911775 TI - Finding malignant findings from radiological reports using medical attributes and syntactic information. AB - Radiology reports are written primarily in natural language. Automated extraction of malignant findings from narrative reports is an important technique for clinical support or alert generation for physicians. This paper proposes a method for automatically extracting malignant findings from narrative radiological reports written in Japanese. First, sentences are parsed and a medical attribute of each phrase is determined. Next, sub-trees related to radiological findings are extracted from a dependency tree using medical attributes. Finally, the malignant findings in each sub tree are extracted with their positive or negative assertions, each of which is determined by the multiplication of pos/neg signs along a path in a sub-tree. The recall and precision for the extraction of malignant findings with their positive or negative assertions were 76% and 91% respectively. The experimental results showed the validity of the proposed method for extracting malignant findings with correct assertions. PMID- 17911776 TI - A normalized lexical lookup approach to identifying UMLS concepts in free text. AB - The National Library of Medicine has developed a tool to identify medical concepts from the Unified Medical Language System in free text. This tool - MetaMap (and its java version MMTx) has been used extensively for biomedical text mining applications. We have developed a module for MetaMap which has a high performance in terms of processing speed. We evaluated our module independently against MetaMap for the task of identifying UMLS concepts in free text clinical radiology reports. A set of 1000 sentences from neuro-radiology reports were collected and processed using our technique and the MMTx Program. An evaluation showed that our technique was able to identify 91% of the concepts found by MMTx in 14% of the time taken by MMTx. An error analysis showed that the missing concepts were largely those which were not direct lexical matches but inferential matches of multiple concepts. Our method also identified multi-phrase concepts which MMTx failed to identify. We suggest that this module be implemented as an option in MMTx for real-time text mining applications where single concepts found in the UMLS need to be identified. PMID- 17911777 TI - Extracting subject demographic information from abstracts of randomized clinical trial reports. AB - In order to make more informed healthcare decisions, consumers need information systems that deliver accurate and reliable information about their illnesses and potential treatments. Reports of randomized clinical trials (RCTs) provide reliable medical evidence about the efficacy of treatments. Current methods to access, search for, and retrieve RCTs are keyword-based, time-consuming, and suffer from poor precision. Personalized semantic search and medical evidence summarization aim to solve this problem. The performance of these approaches may improve if they have access to study subject descriptors (e.g. age, gender, and ethnicity), trial sizes, and diseases/symptoms studied. We have developed a novel method to automatically extract such subject demographic information from RCT abstracts. We used text classification augmented with a Hidden Markov Model to identify sentences containing subject demographics, and subsequently these sentences were parsed using Natural Language Processing techniques to extract relevant information. Our results show accuracy levels of 82.5%, 92.5%, and 92.0% for extraction of subject descriptors, trial sizes, and diseases/symptoms descriptors respectively. PMID- 17911778 TI - Coupling ontology driven semantic representation with multilingual natural language generation for tuning international terminologies. AB - OBJECTIVES: The importance of clinical communication between providers, consumers and others, as well as the requisite for computer interoperability, strengthens the need for sharing common accepted terminologies. Under the directives of the World Health Organization (WHO), an approach is currently being conducted in Australia to adopt a standardized terminology for medical procedures that is intended to become an international reference. METHOD: In order to achieve such a standard, a collaborative approach is adopted, in line with the successful experiment conducted for the development of the new French coding system CCAM. Different coding centres are involved in setting up a semantic representation of each term using a formal ontological structure expressed through a logic-based representation language. From this language-independent representation, multilingual natural language generation (NLG) is performed to produce noun phrases in various languages that are further compared for consistency with the original terms. RESULTS: Outcomes are presented for the assessment of the International Classification of Health Interventions (ICHI) and its translation into Portuguese. The initial results clearly emphasize the feasibility and cost effectiveness of the proposed method for handling both a different classification and an additional language. CONCLUSION: NLG tools, based on ontology driven semantic representation, facilitate the discovery of ambiguous and inconsistent terms, and, as such, should be promoted for establishing coherent international terminologies. PMID- 17911779 TI - Biomedical knowledge discovery with topological constraints modeling in Bayesian networks: a preliminary report. AB - Serving as exploratory data analysis tools, Bayesian networks (BNs) can be automatically learned from data to compactly model direct dependency relationships among the variables in a domain. A major challenge in BN learning is to effectively represent and incorporate domain knowledge in the learning process to improve its efficiency and accuracy. In this paper, we examine two types of domain knowledge representation in BNs: matrix and rule. We develop a set of consistency checking mechanisms for the representations and describe their applications in BN learning. Empirical results from the canonical Asia network example show that topological constraints, especially those imposed on the undirected links in the corresponding completed partially directed acyclic graph (CPDAG) of the learned BN, are particularly useful. Preliminary experiments on a real-life coronary artery disease dataset show that both efficiency and accuracy can be improved with the proposed methodology. The bootstrap approach adopted in the BN learning process with topological constraints also highlights the set of the learned links with high significance, which can in turn prompt further exploration of the actual relationships involved. PMID- 17911780 TI - Automatic infection detection system. AB - An infected person may be contagious already before the first symptoms appear. This person can, in the period of disease evolution, infect several associated citizens before consulting a general practitioner (GP). Early detection of contagion is therefore important to prevent spreading of diseases. The Automatic Infection Detection (AID) System faces this problem through investigating the hypothesis that the blood glucose (BG) level increases when a person is infected. The first objective of the prototyped version of the AID system was to identify possible BG elevations in the incubation time that could be related to the spread of infectious diseases. To do this, we monitored two groups of people, with and without diabetes mellitus. The AID system analyzed the results and we were able to detect two cases of infection during the study period. The time of detection occurred simultaneous or near the time of onset of symptoms. The detection did not occur earlier for a number of reasons. The most likely one is that the evolution process of an infectious disease is both complicated and involves the immune system and several organs in the body. The investigation with regard to isolating the key relations is therefore considered as a very complex study. Nevertheless, the AID system managed to detect the infection much earlier than what is possible with today's early warning systems for infectious diseases. PMID- 17911781 TI - Risk stratification for LDL cholesterol using induction technique. AB - We identified the combined patterns of LDL cholesterol risk factors including biometric, environmental and genetic factors using induction technique. In this hospital based cardiovascular genome study of Korean men and women, we found that CART (classification and regression tree) was a better method to predict LDL cholesterol compared to the regression method. The CART had a better prediction ability than the multiple regression for male and female, respectively. We also identified combined patterns of LDL cholesterol risk factors and segment specific information for LDL cholesterol management using induction rules. The CART method provided more detailed results according to each segmentation and subgroup. In addition, we demonstrated how the CART algorithm could be used in risk assessment and target segmentation of LDL cholesterol management. PMID- 17911782 TI - Results from data mining in a radiology department: the relevance of data quality. AB - This work is part of an ongoing effort to examine and improve clinical workflows in radiology. Classical workflow analysis is time consuming and expensive. Here we present a purely data-driven approach using data mining techniques to detect causes for poor data quality and areas with poor workflow performance. Data has been taken from a operational RIS system. We defined a set of four key indicators for both data quality and workflow performance. Using several mining techniques such as cluster analysis and correlation tests we were able to detect interesting effects regarding data quality and an abnormality in the workflow for some organizational units of the examined radiology departments. We conclude that data driven data mining approaches may act as a valuable tool to support workflow analysis and can narrow down the problem space for a manual on-site workflow analysis. This can save time and effort and leads to less strain for clinicians and workflow analysts during interviews. PMID- 17911783 TI - Evaluating learning models with transitions of human interests based on objective rule evaluation indices. AB - This paper presents a method to support the evaluation procedure of a data mining process using human-system interaction. The post-processing of mined results is one of the key factors for successful data mining process. However, it is difficult for human experts to completely evaluate several thousands of rules from a large dataset containing noise. We have designed a method based on objective rule evaluation indices to support the rule evaluation procedure; the indices are calculated to evaluate each if-then rule mathematically. We have evaluated five representative learning algorithms to construct rule evaluation models of the actual data mining results from a chronic hepatitis data set. Further, we discuss the relationship between the transitions of the subjective criterion of a medical expert and the performances of the rule evaluation models. PMID- 17911784 TI - An automated data pattern translation process for medical data mining. AB - This paper describes the development and application of a medical pattern evaluation methodology. The ADAPT process aims to facilitate the increased use of data mining technologies in medicine by providing a method of translating the language of data mining outputs into that of the medical domain. The research behind this work was developed in response to issues raised both in the literature and in public forums regarding the difficulty of adapting data mining technologies for the unique needs of the medical domain. This paper also responds to the need for automated systems to reflect the methodologies and processes applied in the management and analysis of medical data. The solution presented here shows promise as a method of providing a bridge between traditional data mining and the needs of medical domain users. PMID- 17911785 TI - A data mining approach to analyze non-compliance with a guideline for the treatment of breast cancer. AB - Postmastectomy radiotherapy (PMRT) is prescribed in order to reduce the local recurrence of breast cancer and improve overall survival. A guideline supports the trade-off between benefits and adverse effects of PMRT. However, this guideline is not always followed in practice. This study tries to find a method for revealing patterns of non-compliance between the actual treatment and the PMRT guideline. Data from breast cancer patients admitted to Linkoping University Hospital between 1990 and 2000 were analyzed in this study. Cases that were not treated in accordance with the guideline were selected and analyzed by decision tree induction (DTI). Thereafter, four resulting rules, as representations for groups of patients, were compared to the guideline. Finding patterns of non compliance with guidelines by means of rules can be an appropriate alternative to manual methods, i.e. a case-by-case comparison when studying very large datasets. The resulting rules can be used in a knowledge base of a guideline-based decision support system to alert when inconsistencies with the guidelines may appear. PMID- 17911786 TI - Bayesian networks for multivariate data analysis and prognostic modelling in cardiac surgery. AB - Prognostic models are tools to predict the outcome of disease and disease treatment. These models are traditionally built with supervised machine learning techniques, and consider prognosis as a static, one-shot activity. This paper presents a new type of prognostic model that builds on the Bayesian network methodology that implements a dynamic, process-oriented view on prognosis. In contrast to traditional prognostic models, prognostic Bayesian networks explicate the scenarios that lead to disease outcomes, and can be used to update predictions when new information becomes available. A recursive data analysis strategy for inducing prognostic Bayesian networks from medical data is presented, and applied to data from the field of cardiac surgery. The resulting model outperformed a model that was constructed with off-the-shelf Bayesian network learning software, and had similar performance as class probability trees. PMID- 17911787 TI - Role of syndromic management using dynamic machine learning in future of e-Health in Pakistan. AB - Sexually Transmitted Diseases (STDs) constitute important primary health issues in Pakistan which face inadequacy of resources required in early detection and investigative procedures for their diagnosis and treatment. Syndromic approach to management of STDs is based on the identification of a consistent group of symptoms and syndromes to classify the exact disease or infection beforehand, so that further investigations are sought for based on these initial criteria. This paper envisions the results based on two different approaches: Human and Artificial Intelligence (AI) along with some examples of on-going usage e of Artificial Intelligence in Medicine. Pakistan is in an early stage regarding the use of informatics in health care for sustainable health system but is also under international obligation to adapt it & WHO EMRO has developed an e-Health plan for the member countries including Pakistan. The paper presents an informatics application model for a very common but important problem & development of such e health applications, as starting point, will certainly have positive impact the future of development of e-Health in Pakistan. PMID- 17911788 TI - Combining lexical and semantic methods of inter-terminology mapping using the UMLS. AB - The need for inter-terminology mapping is constantly increasing with the growth in the volume of electronically captured biomedical data and the demand to re-use the same data for secondary purposes. Using the UMLS as a knowledge base, semantically-based and lexically-based mappings were generated from SNOMED CT to ICD9CM terms and compared to a gold standard. Semantic mapping performed better than lexical mapping in terms of coverage, recall and precision. As the two mapping methods are orthogonal, the two sets of mappings can be used to validate and enhance each other. A method of combining the mappings based on the precision level of sub-categories in each method was derived. The combined method outperformed both methods, achieving coverage of 91%, recall of 43% and precision of 27%. It is also possible to customize the method of combination to optimize performance according to the task at hand. PMID- 17911789 TI - Biomedical vocabularies--the demand for differentiation. AB - The need of biomedical vocabularies is well known for various tasks, e.g., supporting structured data entry, decision support and electronic data exchange as well as retrieval and statistical evaluation of the data. Due to a considerable diversity of artifacts like interface terminologies, classifications and thesauri it seems to be reasonable to demand for a massive reduction and, finally, to end up with just one unique "multi-purpose world terminology". Concept-based reference terminologies like SNOMED CT might be candidates for that idea. Unfortunately, the above mentioned kinds of vocabularies cannot be replaced because of their specific purpose-dependent nature. Their mutual distinctive characteristics are outlined and compared with inherent purpose-less concept systems that are based on formal approaches like description logics. For supporting interoperability the different kinds of purpose-dependent vocabularies can and should be improved by mappings to machine-processible reference terminologies. As a side-effect, this paper may contribute to the meta terminology in this field of medical terminology. PMID- 17911790 TI - Development of a taxonomy for health information technology. AB - Taxonomies provide schemas to help classify entities and define the relationships between them. Early computing enabled the development of ontologies and Medical Subject Headings (MeSH), the first modern classification of medical terminology as applied to medical literature. Later developments, such as MEDLINE, expanded MeSH to include a number of medical informatics terms. However, a lack of specificity in MeSH and other existing informatics taxonomies for terminology used to describe the growing field of health information technology (health IT) created the need for the development of a specialized taxonomy. Experts associated with the Agency for Healthcare Research and Quality (AHRQs) National Resource Center for Health Information Technology (NRC) created and evaluated a taxonomy for health IT, to enable users of a public health IT Web site to efficiently identify resources within an online, searchable repository. PMID- 17911791 TI - A practical approach to advanced terminology services in health information systems. AB - As the medical informatics field evolves, new functions appear as the focus of interest; a more advanced management of terminology is one of them. Using comprehensive and detailed terminology to represent clinical rules in computer systems, associated with patient information, would allow clinical software to provide patient specific recommendations or alerts. In order to uniform data collection through our HIS, and lay the foundations for future clinical decision support systems, we decided to move from our previous classification-based medical record into new terminology services built around Snomed CT, Spanish Language Version. This paper describes the characteristics of our Terminology Server. The most important achievements of our new terminology system are the centralization of knowledge representation, using a much more detailed terminology system. Clinical data entered at any place of the institution and level of care, is represented uniformly through the whole health information system. PMID- 17911792 TI - Toward the interoperability of HL7 v3 and SNOMED CT: a case study modeling mobile clinical treatment. AB - Semantic interoperability in healthcare can be achieved by a tighter coupling of terminology and HL7 message models. In this paper, we highlight the difficulty of achieving this goal, but show how it can become attainable by basing HL7 message models on SNOMED CT concepts and relationships. We then demonstrate how this methodology has been applied to a set of clinical observations for use in the ePOC project, and discuss our findings. PMID- 17911793 TI - A web-based SNOMED CT browser: distributed and real-time use of SNOMED CT during the clinical research process. AB - To facilitate the use of standard terminologies in clinical research data collection, members of the Rare Disease Clinical Research Network have developed tools to support study investigators and research staff to code clinical research data using SNOMED CT at the point of research. This tool is customized to help the user find appropriate SNOMED CT concepts quickly, and has implications for the successful implementation of data standards to facilitate high quality research data. This paper gives an overview of an automated tool for accessing, searching, and navigating SNOMED CT real-time, at distributed and remote clinical study locations. Also, the features of the tool that enable complete data are presented, as well as possible metrics for evaluation in terms of compliance, consistency, and reliability of coding. PMID- 17911794 TI - Using SNOMED CT as a reference terminology to cross map two highly pre coordinated classification systems. AB - We hypothesized that SNOMED CT, a granular formal reference terminology, could be used to assist in the creation of a valid crosswalk between two administrative classifications: the International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) and the U.S. Veterans Benefits Administration (VBA) disability code set. To establish a baseline, we created an ICD-9-CM terminology server and directly mapped textual descriptions of the VBA disability codes to ICD-9-CM. We next mapped ICD-9-CM and the VBA Disability codes to SNOMED CT. The SNOMED CT mappings were matched across classification systems and terms from related concepts were displayed for an expert coder's review. We report the rate of direct ICD-9-CM to VBA Disability Code mapping (26%), the eventual success of the SNOMED CT based crosswalk (95%) and the rate at which the reviewer had to add codes to complete the mapping (99%). The method using the SNOMED CT crosswalk provided significantly better coverage than the ICD-9-CM direct mapping alone (Pearson Chi Square test; p<0.001). We conclude that SNOMED CT can be a useful adjunct to direct mapping between administrative classifications. PMID- 17911795 TI - Practical issues in using SNOMED CT as a reference terminology. AB - SNOMED CT was created by the merger of SNOMED RT (Reference Terminology) and Read Codes Version 3 (also known as Clinical Terms Version 3). SNOMED CT is considered to be among the most extensive and comprehensive biomedical vocabularies available today. It is considered for use as the Reference Terminology of various institutions. We review the adequacy of SNOMED CT as a Reference Terminology and discuss the issues in its use as such. We discuss issues with content coverage of various clinical domains, data integrity and validity, and the update frequency of SNOMED CT, and why SNOMED CT alone is not adequate to serve as the Reference Terminology of a healthcare organization. PMID- 17911796 TI - Statistical selector of the best multiple ICD-coding method. AB - The International Classification of Diseases 10th version (ICD-10) is one of the standard and most important disease classifications. Since computerized ICD-10 coding systems have drawn a great deal of attention in the medical field, a great number of different coding systems have been proposed. This paper proposes a hybrid architecture of different coding systems. First, given an input disease name, three coding systems output codes with their confidence scores. A C4.5 based system selector then selects the best output by using both input statistics and the confidence score from each system. The experimental results demonstrated that the selector significantly boosts the overall performance (+3.4 points). PMID- 17911797 TI - Creation and evaluation of a terminology server for the interactive coding of discharge summaries. AB - Free text entry versus structured data has been proposed as models in data entry in health information systems. A new user interface was developed with the objective of improving data capture. It also implemented a modification of the discharge summary data entry user interface that allowed the selection of already coded terms from a local terminology in the context of an inpatient electronic medical record. This software interacts online with a terminology server to provide feedback on data entry to clinical users in order to automatically code data. To evaluate the impact of this new software, we measured user satisfaction and the impact on autocodification rate. The new system had good acceptance from the users who ranked it high using QUIS (Questionnaire for User Interaction Satisfaction) and the auto codification rate improved from 61.5% to 88.39%. PMID- 17911798 TI - Thesaurus anomaly detection by user action monitoring. AB - The construction and maintenance of a medical thesaurus is a non-trivial task, due to the inherent complexity of a proper medical terminology. We present a methodology for transaction-based anomaly detection in the process of thesaurus maintenance. Our experiences are based on lexicographic work with the MorphoSaurus lexicons, which are the basis for a mono- and cross-lingual biomedical information retrieval system. Any "edit"or "delete" actions within these lexicons that undo an action defined earlier were defined as anomalous. We identify four types of such anomalies. We also analyzed to which extent the anomalous lexicon entries had been detected by an alternative, corpus-based approach. PMID- 17911799 TI - Harmonizing clinical terminologies: driving interoperability in healthcare. AB - Internationally, there are countless initiatives to build National Healthcare Information Networks (NHIN) that electronically interconnect healthcare organizations by enhancing and integrating current information technology (IT) capabilities. The realization of such NHINs will enable the simple and immediate exchange of appropriate and vital clinical data among participating organizations. In order for institutions to accurately and automatically exchange information, the electronic clinical documents must make use of established clinical codes, such as those of SNOMED-CT, LOINC and ICD-9 CM. However, there does not exist one universally accepted coding scheme that encapsulates all pertinent clinical information for the purposes of patient care, clinical research and population heatlh reporting. In this paper, we propose a combination of methods and standards that target the harmonization of clinical terminologies and encourage sustainable, interoperable infrastructure for healthcare. PMID- 17911800 TI - A new machine learning classifier for high dimensional healthcare data. AB - Data sets with many discrete variables and relatively few cases arise in health care, commerce, information security, and many other domains. Learning effective and efficient prediction models from such data sets is a challenging task. In this paper, we propose a new approach that combines Metaheuristic search and Bayesian Networks to learn a graphical Markov Blanket-based classifier from data. The Tabu Search enhanced Markov Blanket (TS/MB) procedure is based on the use of restricted neighborhoods in a general Bayesian Network constrained by the Markov condition, called Markov Blanket Neighborhoods. Computational results from two real world healthcare data sets indicate that the TS/MB procedure converges fast and is able to find a parsimonious model with substantially fewer predictor variables than in the full data set. Furthermore, it has comparable or better prediction performance when compared against several machine learning methods, and provides insight into possible causal relations among the variables. PMID- 17911801 TI - Structuring of free-text diagnostic report. AB - PURPOSE: It is useful to convert free-text diagnostic reports into structured diagnostic reports by semantic analysis for the secondary investigation of their contents. In this study, we propose a system in which description units are automatically extracted to create structured text reports and we evaluated its usefulness. METHODS: We defined the rules to create description units and developed the system that can automatically extract these description units from free-text diagnostic reports. We applied this system to reports of cerebral perfusion scintigrams and obtained 5 dictionaries of description units, increasing the number of scintigrams from 100 to 500 in increments of 100. Each dictionary was used to analyze another 100 scintigrams. The results obtained using each dictionary were compared with the results of physicians' interpretation. RESULTS: The recall rate of this system to the physicians' interpretation increased when correlated with the number of scintigrams but with 300 cases was almost saturated at 85%. CONCLUSION: We propose a semantic analysis system and show its usefulness in the semantic evaluation of the reports of cerebral perfusion scintigrams. PMID- 17911802 TI - Semantic issues in integrating data from different models to achieve data interoperability. AB - Matching clinical data to codes in controlled terminologies is the first step towards achieving standardisation of data for safe and accurate data interoperability. The MoST automated system was used to generate a list of candidate SNOMED CT code mappings. The paper discusses the semantic issues which arose when generating lexical and semantic matches of terms from the archetype model to relevant SNOMED codes. It also discusses some of the solutions that were developed to address the issues. The aim of the paper is to highlight the need to be flexible when integrating data from two separate models. However, the paper also stresses that the context and semantics of the data in either model should be taken into consideration at all times to increase the chances of true positives and reduce the occurrence of false negatives. PMID- 17911804 TI - Comparing medical code usage with the compression-based dissimilarity measure. AB - It is well known that medical coding practice is inconsistent and that differences in usage may exist even at the institutional level. In this paper we introduce a novel method for investigating code usage patterns in clinical documentation corpora. By applying the Compression-based Dissimilarity Measure to calculate similarities between encounter notes, we find that certain notes can be associated with a number of different classifications and that a given classification code can be documented in fundamentally different ways. The effect is that some notes need to be understood in the context of the classification code, a finding which has implications for data mining or information extraction tasks. In addition, the method opens for a number of interesting application areas that include highlighting code use anomalies, measuring how coding practice changes over time, comparing code usage across institutions, and, perhaps most importantly, provide valuable feedback to developers of classification coding systems. PMID- 17911803 TI - Medication reconciliation using natural language processing and controlled terminologies. AB - Medication reconciliation (MR) is a process that seeks to assure that the medications a patient is supposed to take are the same as what they are actually taking. We have developed a method in which medication information (consisting of both coded data and narrative text) is extracted from twelve sources from two clinical information systems and assembled into a chronological sequence of medication history, plans, and orders that correspond to periods before, during and after a hospital admission. We use natural language processing, a controlled terminology, and a medication classification system to create matrices that can be used to determine the initiation, changes and discontinuation of medications over time. We applied the process to a set of 17 patient records and successfully abstracted and summarized the medication data. This approach has implications for efforts to improve medication history-taking, order entry, and automated auditing of patient records for quality assurance. PMID- 17911805 TI - A scale-free network view of the UMLS to learn terminology translations. AB - The UMLS Metathesaurus belongs to the class of scale-free networks with few concept hubs possessing a large number of relationships. The hubs provide useful links between the concepts from disparate terminologies in the UMLS; however, they also exponentially increase the number of possible transitive cross terminology paths. Towards the goal of using machine learning to rank cross terminology translations, we propose a traversal algorithm that exploits the scale-free property of the UMLS to reduce the number of candidate translations. We characterize the concept hubs into "informational" and "noisy" concept hubs and provide an automated method to detect them. Using gold standard mappings from SNOMED-CT to ICD9CM, we found an average 20-fold reduction in the number of candidate mappings while achieving comparable recall and ranking results. A hub driven traversal strategy provides a promising approach to generate high quality cross-terminology translations from the UMLS. PMID- 17911806 TI - Assigning categorical information to Japanese medical terms using MeSH and MEDLINE. AB - This paper reports on the assigning of MeSH (Medical Subject Headings) categories to Japanese terms in an English-Japanese dictionary using the titles and abstracts of articles indexed in MEDLINE. In a previous study, 30,000 of 80,000 terms in the dictionary were mapped to MeSH terms by normalized comparison. It was reasoned that if the remaining dictionary terms appeared in MEDLINE-indexed articles that are indexed using MeSH terms, then relevancies between the dictionary terms and MeSH terms could be calculated, and thus MeSH categories assigned. This study compares two approaches for calculating the weight matrix. One is the TF*IDF method and the other uses the inner product of two weight matrices. About 20,000 additional dictionary terms were identified in MEDLINE indexed articles published between 2000 and 2004. The precision and recall of these algorithms were evaluated separately for MeSH terms and non-MeSH terms. Unfortunately, the precision and recall of the algorithms was not good, but this method will help with manual assignment of MeSH categories to dictionary terms. PMID- 17911807 TI - PharmARTS: terminology web services for drug safety data coding and retrieval. AB - MedDRA and WHO-ART are the terminologies used to encode drug safety reports. The standardisation achieved with these terminologies facilitates: 1) The sharing of safety databases; 2) Data mining for the continuous reassessment of benefit-risk ratio at national or international level or in the pharmaceutical industry. There is some debate about the capacity of these terminologies for retrieving case reports related to similar medical conditions. We have developed a resource that allows grouping similar medical conditions more effectively than WHO-ART and MedDRA. We describe here a software tool facilitating the use of this terminological resource thanks to an RDF framework with support for RDF Schema inferencing and querying. This tool eases coding and data retrieval in drug safety. PMID- 17911808 TI - Machine learning approach for automatic quality criteria detection of health web pages. AB - The number of medical websites is constantly growing [1]. Owing to the open nature of the Web, the reliability of information available on the Web is uneven. Internet users are overwhelmed by the quantity of information available on the Web. The situation is even more critical in the medical area, as the content proposed by health websites can have a direct impact on the users' well being. One way to control the reliability of health websites is to assess their quality and to make this assessment available to users. The HON Foundation has defined a set of eight ethical principles. HON's experts are working in order to manually define whether a given website complies with s the required principles. As the number of medical websites is constantly growing, manual expertise becomes insufficient and automatic systems should be used in order to help medical experts. In this paper we present the design and the evaluation of an automatic system conceived for the categorisation of medical and health documents according to he HONcode ethical principles. A first evaluation shows promising results. Currently the system shows 0.78 micro precision and 0.73 F-measure, with 0.06 errors. PMID- 17911809 TI - Using discourse analysis to improve text categorization in MEDLINE. AB - PROBLEM: Automatic keyword assignment has been largely studied in medical informatics in the context of the MEDLINE database, both for helping search in MEDLINE and in order to provide an indicative "gist" of the content of an article. Automatic assignment of Medical Subject Headings (MeSH), which is formally an automatic text categorization task, has been proposed using different methods or combination of methods, including machine learning (naive Bayes, neural networks..), linguistically-motivated methods (syntactic parsing, semantic tagging, or information retrieval. METHODS: In the present study, we propose to evaluate the impact of the argumentative structures of scientific articles to improve the categorization effectiveness of a categorizer, which combines linguistically-motivated and information retrieval methods. Our argumentative categorizer, which uses representation levels inherited from the field of discourse analysis, is able to classify sentences of an abstract in four classes: PURPOSE; METHODS; RESULTS and CONCLUSION. For the evaluation, the OHSUMED collection, a sample of MEDLINE, is used as a benchmark. For each abstract in the collection, the result of the argumentative classifier, i.e. the labeling of each sentence with an argumentative class, is used to modify the original ranking of the MeSH categorizer. RESULTS: The most effective combination (+2%, p<0.003) strongly overweights the METHODS section and moderately the RESULTS and CONCLUSION section. CONCLUSION: Although modest, the improvement brought by argumentative features for text categorization confirms that discourse analysis methods could benefit text mining in scientific digital libraries. PMID- 17911810 TI - A comparison of impact factor, clinical query filters, and pattern recognition query filters in terms of sensitivity to topic. AB - Evaluating journal quality and finding high-quality articles in the biomedical literature are challenging information retrieval tasks. The most widely used method for journal evaluation is impact factor, while novel approaches for finding articles are PubMed's clinical query filters and machine learning-based filter models. The related literature has focused on the average behavior of these methods over all topics. The present study evaluates the variability of these approaches for different topics. We find that impact factor and clinical query filters are unstable for different topics while a topic-specific impact factor and machine learning-based filter models appear more robust. Thus when using the less stable methods for a specific topic, researchers should realize that their performance may diverge from expected average performance. Better yet, the more stable methods should be preferred whenever applicable. PMID- 17911811 TI - A method for defining a journal subset for a clinical discipline using the bibliographies of systematic reviews. AB - BACKGROUND: Searching for best evidence for clinical decisions in large biomedical databases is problematic because advances in health care practice that are ready for application are but a very dilute constituent in a much larger pool of biomedical literature. Sensitive search strategies have been developed to help alleviate this problem but search precision is still generally low. If "virtual journal subsets" that are likely to include all relevant articles can be defined for clinical discipline areas or disease content areas this will likely improve search precision. OBJECTIVE: To determine whether studies cited in systematic literature reviews can define a journal subset for a given clinical discipline. DESIGN: Survey of the primary studies included in systematic reviews that are relevant to the clinical discipline of nephrology. METHODS: Four data sources were searched to identify systematic reviews relevant to clinical nephrology: the Cochrane Database of Systematic Reviews, McMaster PLUS (Premium LiteratUre Service), MEDLINE, and the Renal Health Library. Three research assistants recorded data pertinent to each of the included primary studies. RESULTS: 195 systematic reviews relevant to nephrology were defined and the 2,779 unique original articles they cited were concentrated in 466 journals, with 90% of the articles in 217 titles. This journal subset can be stored online and used when searching the large biomedical data-bases such as MEDLINE. CONCLUSION: The bibliographies of systematic reviews can be used to define a journal subset for a clinical discipline area. PMID- 17911812 TI - Aequus communis sententia: defining levels of interoperability. AB - Interoperability is a common understanding of the meaning of data between a sending and receiving computer system [1]. The level of interoperability required varies with application needs. The specification of data in enough detail to create a common shared meaning between organizations is a complex task as systems work within organizational and human factors contexts as well as having specific technical requirements. Aequus communis sententia translates from Latin to the "level of common meaning." In this manuscript, we define an Ontology of Interoperability. The scale asks reviewers of a specification to define it's level in terms of Syntactic, Semantic and Pragmatic Interoperability. We tested the scale by having five medical Informaticians rate a set of ANSI standard specifications and we report the inter-rater variability of the interoperability rating scheme. We learned that some elements of the scale presented more difficulty for our reviewers and based on our findings we present a final version of the interoperability scale in our discussion. Our interoperability rating ontology has high inter-rater reliability and is a relatively simple mechanism for comparing the levels of interoperability afforded by different specifications or the same specification over multiple versions. PMID- 17911813 TI - What's in a code? Towards a formal account of the relation of ontologies and coding systems. AB - Terminologies are increasingly based on "ontologies" developed in description logics and related languages such as the new Web Ontology Language, OWL. The use of description logic has been expected to reduce ambiguity and make it easier determine logical equivalence, deal with negation, and specify EHRs. However, this promise has not been fully realised: in part because early description logics were relatively inexpressive, in part, because the relation between coding systems, EHRs, and ontologies expressed in description logics has not been fully understood. This paper presents a unifying approach using the expressive formalisms available in the latest version of OWL, OWL 1.1. PMID- 17911814 TI - A road from health care classifications and coding systems to biomedical ontology: the CEN categorial structure for terminologies of human anatomy: Catanat. AB - There is an increasing need for updated and harmonised health care classifications and coding systems to allow international comparisons and cooperation for instance for population based WHO indicators, Electronic Health Record safety, trans border migration of population, case mix and procedure payment. It is not feasible to propose a standardisation of the linguistic expressiveness of different health care professionals and of the citizens. Natural language expressions show inconsistencies and ambiguities as assessed by biomedical ontology driven tools. In order to build a road towards standardisation the European Standard Body CEN has stated that it was not possible to convince the different European member states using different national languages to agree on a reference clinical terminology as Snomed-CT or to standardise a detailed language independent biomedical ontology. It has developed since 1990 an approach named categorial structure as a step standardising only the terminologies model structure. The categorial structure for terminologies of human anatomy currently in the phase of final approval is presented as a methodology for bridging between classifications and coding systems and biomedical ontology on the way to semantic interoperability between different languages, legacies and goals. PMID- 17911815 TI - The nodes focusing tool for clinical course data of hypergraph structure in the ontological framework CSX output from POMR-based EMR system. AB - Knowledge management is one of the significant issues in this century. In medical informatics, the concept of ontology is an important current topic. In addition, there is great interest in knowledge acquisition from clinical data. When doctors use clinical information systems (CIS), their operation implicitly represents, to some extent, their thinking processes with clear reasons and goals according to their intent. If we can capture such resources in an appropriate representation, considerable empirical knowledge could be utilized in various research fields. With this prospect, the authors built an experimental CIS and a nodes-focusing tool for abstracting a summary of the clinical course being examined. The information model is based on a thinking process model, and data are represented in the ontological framework CSX. Then we showed the knowledge-abstracting procedures with this tool. Some domain experts and intern students showed great interest. The authors thereby concluded that such an empirical knowledge acquisition environment is useful for research, education, and so on. PMID- 17911816 TI - Integrating descriptive, functional and cooperative aspects in a domain ontology to computerize home care charts. AB - To coordinate themselves, home care (HC) professionals use artifacts to keep a mutual understanding on their common activity: lightly structured charts written in natural language. Instead of trying to define a record to capture them, we want to focus on efficient indexing of this information. The use of a Domain Ontology was proposed. This paper explains how we built and implemented it. Three complementary aspects of the HC charts were analyzed (i) functional aspects performed with precise analyses of actual charts; (ii) interoperability aspects with the use of some HL7-RIM standardized descriptions; and (iii) cooperative aspects, with the integration of a cooperation model. We proposed a Domain Ontology to represent the concepts and the relations used in the charts. The implementation was done with Protege; the ontology was built in OWL-DL. The IAnnotate application helps us to index the HC chart with the domain ontology. The next step of the work will use the ontology to reason about the different items of information contained in the charts so that contextual use of them should be envisaged. PMID- 17911817 TI - The ICNP-BaT--a multilingual web-based tool to support the collaborative translation of the International Classification for Nursing Practice (ICNP). AB - To ease and speed up the translation of the ICNP version 1 into the German language a web service was developed to support the collaborative work of all Austrian, Swiss, and German translators and subsequently of the evaluators of the resultant translation. The web service does help to support a modified Delphi technique. Since the web service is multilingual by design it can facilitate the translation of the ICNP into other languages as well. The process chosen can be adopted by other projects involved in translating terminologies. PMID- 17911818 TI - Ontology based modeling of pandemic simulation scenarios. AB - Computer-based simulation of influenza outbreaks in local communities can help researchers, epidemiologists, and decision makers better understand the impact of the community structure on the reproduction rate of disease, and the relative benefits of different types of prevention and interventions. The goal of scenario modeling is to develop a description of scenario components, such as the disease, the community, and interventions. An ontology-based representation of the scenario model together with a modeling tool, which is based on an extension to Protege, assist scenario developers in formulating simulation specifications. This approach allows the exploration of new ideas by rapidly formulating and reconstructing scenarios from novel components. PMID- 17911819 TI - An ontology-based model of clinical information. AB - In this paper we describe a model of clinical information designed to make health information systems properly interoperable and safely computable. The model is a response to a number of categories of requirements, ranging from the semantic to the performance of software at runtime. We argue that the starting point of a successful model must be an ontological analysis of the process of clinical care delivery, seen as a scientific problem-solving process. From this approach we develop a classification of types of clinical information called the Clinical Investigator Record (CIR) ontology. PMID- 17911820 TI - Creation of a local interface terminology to SNOMED CT. AB - This paper describes the steps followed in the creation of a local Interface Terminology to SNOMED CT (as reference terminology) with a strong focus on user acceptability. The resulting list of terms is used for clinical data input by physicians and nurses at the Hospital Italiano in Buenos Aires, Argentina. Description includes data model, mappings to SNOMED CT and classifications, subsets definitions and extensibility mechanisms. The Interface Terminology is currently used in the recording of diagnosis and procedures in inpatient discharge summaries and its coverage is improving from user feedback. Its current size is 24,800 concepts, 67% of them needed post-coordination for appropriate semantic representation, due to a very flexible policy that allows the use of any number of modifiers on concepts. PMID- 17911821 TI - A feasibility study on clinical templates for the National Health Service in Scotland. AB - There is growing interest in the development of standards which structure information round discrete clinical concepts, in a way that supports system development and interoperability. Most notable are the openEHR Archetypes and Templates, and HL7 Templates. A project is described which explored the potential for these to engage and support clinical practitioners in developing clinical information standards in an open and accessible way. The project followed three phases: professional development, including networking for content development; library implementation, including modelling and processes for managing content and relating to information standards; and implementation in working systems in a manner that is professionally acceptable. The project findings are encouraging although there remain some important issues to be explored in further work. The topic has now emerged as an important area of standards development, and a useful focus for international cooperation. PMID- 17911822 TI - Keeping up with changing source system terms in a local health information infrastructure: running to stand still. AB - Keeping up with changes in source system terms in a local health information infrastructure requires substantial effort. I developed a program to assist us that returns candidate mappings based on string similarities between newly encountered source test names, existing source test names, and our master dictionary term names. I evaluated this program's performance in identifying correct mappings through a retrospective study of term mappings to our master dictionary from four radiology systems. For source terms created after the initial system integration, the semi-automated mapping program identified correct mappings for 76.3% of terms from all systems. Overall, the program correctly identified mappings for 45.6% of all terms by exact string match to an existing term. The program identified correct mappings for 36.9% of the terms without an exact string match by string comparison to existing source terms, and for 54.4% of the remaining unmapped terms by string comparison directly to master dictionary terms. Because managing vocabulary mappings is resource-intensive, accurate automated tools can help reduce the effort required for ongoing health information exchange among disparate systems. PMID- 17911823 TI - The role of local terminologies in electronic health records. The HEGP experience. AB - Despite decades of work, there is no universally accepted standard medical terminology and no generally usable terminological tools have yet emerged. The local dictionary of concepts of the Georges Pompidou European Hospital (HEGP) is a Terminological System (TS) designed to support clinical data entry. It covers 93 data entry forms and contains definitions and synonyms of more than 5000 concepts, sometimes linked to reference terminologies such as ICD-10. In this article, we evaluate to which extend SNOMED CT could fully replace or rather be mapped to the local terminology system. We first describe the local dictionary of concepts of HEGP according to some published TS characterization framework. Then we discuss the specific role that a local terminology system plays with regards to reference terminologies. PMID- 17911824 TI - Ontology-based knowledge base model construction-OntoKBCF. AB - Semantic web technologies are used in the construction of a bio-health knowledge base model, which, when coupled with an Electronic Health Record (EHR), is to be used by clinicians. Specifically, this ontology provides the basis for a domain knowledge resource that attempts to bridge biological and clinical information. The prototype is focused on a Cystic Fibrosis exemplar, and the content of the model includes: Cochrane reviews; a time-oriented description; gene therapy; and the most common cystic fibrosis gene mutations. The facts within the model range from nucleo-base mutation and amino acid change to clinical phenotype. The knowledge is represented by layers from the micro level to the macro level. Here, emphasis is placed upon the details between levels (i.e., the vertical axis) and these are made available to bridge the knowledge from different levels. The description of gender, age, mutation and clinical manifestations are clues for matching points within an EHR system. OWL is the ontology representation language used and the output from Protege-OWL is a XML-based file format, which facilitates further application and communication. PMID- 17911825 TI - Integrating biological pathways in disease ontologies. AB - Anatomy, clinical features, etiology, and morphology are the major organizing principles in existing disease ontologies. Assuming that biological pathways (including protein physical interactions, metabolic reactions, regulatory networks) will be in the near future key components in classifications of diseases, we have analyzed how information about pathways can be integrated into disease ontologies. We designed a disease ontology in OWL. SNOMED CT was used to provide the initial disease descriptions. In a second step, we integrated information from the KEGG PATHWAY and the GO annotation data-bases into the disease ontology. In the last step, we analyzed the classification of diseases. For example glioma of brain shares 30 pathways with other cancers, and 19 pathways with Alzheimer's disease. As our knowledge about biological pathways is constantly evolving, this approach can be used for integrating automatically this knowledge in existing ontologies. Thanks to the automatic classification associated with formal ontologies, this approach helps identify physio pathological classes and taxonomic relations in diseases ontologies. It can therefore be used to create new partitions, focusing on pathways, in biomedical ontologies. PMID- 17911826 TI - Reconciliation of ontology and terminology to cope with linguistics. AB - OBJECTIVES: To discuss the relationships between ontologies, terminologies and language in the context of Natural Language Processing (NLP) applications in order to show the negative consequences of confusing them. METHODS: The viewpoints of the terminologist and (computational) linguist are developed separately, and then compared, leading to the presentation of reconciliation among these points of view, with consideration of the role of the ontologist. RESULTS: In order to encourage appropriate usage of terminologies, guidelines are presented advocating the simultaneous publication of pragmatic vocabularies supported by terminological material based on adequate ontological analysis. CONCLUSIONS: Ontologies, terminologies and natural languages each have their own purpose. Ontologies support machine understanding, natural languages support human communication, and terminologies should form the bridge between them. Therefore, future terminology standards should be based on sound ontology and do justice to the diversities in natural languages. Moreover, they should support local vocabularies, in order to be easily adaptable to local needs and practices. PMID- 17911827 TI - SNOMED CT's problem list: ontologists' and logicians' therapy suggestions. AB - After a critical review of the present architecture of SNOMED CT, addressing both logical and ontological issues, we present a roadmap towards an overall improvement of this terminology. In particular, we recommend the following actions: Upper level categories should be rearranged according to a standard upper level ontology. Meta-class like concepts should be identified and removed from the taxonomy. SNOMED concepts denoting (non instantiable) individual entities (e.g. geographical regions) should be kept separate from those concepts that denote (instantiable) types. SNOMED binary relations should be reduced to a set of canonical ones, following existing recommendations. Taxonomies should be cleansed and split into disjoint partitions. The number of full definitions should be increased. Finally, we propose a new approach to modeling part-whole hierarchies, as well as the integration of qualifier relations into the description logic framework. PMID- 17911828 TI - Automatic checking of the correctness of clinical guidelines in GLARE. AB - Representing clinical guidelines is a very complex knowledge-representation task, requiring a lot of expertise and efforts. Nevertheless, guideline representations often contain several kinds of errors. Therefore, checking the well-formedness and correctness of a guideline representation is an important task, which can be drastically improved with the adoption of computer programs. In this paper, we discuss the advanced facilities provided by the GLARE system to assist physicians to produce correct representations of clinical guidelines. PMID- 17911829 TI - Knowledge Zone: a public repository of peer-reviewed biomedical ontologies. AB - Reuse of ontologies is important for achieving better interoperability among health systems and relieving knowledge engineers from the burden of developing ontologies from scratch. Most of the work that aims to facilitate ontology reuse has focused on building ontology libraries that are simple repositories of ontologies or has led to keyword-based search tools that search among ontologies. To our knowledge, there are no operational methodologies that allow users to evaluate ontologies and to compare them in order to choose the most appropriate ontology for their task. In this paper, we present, Knowledge Zone - a Web-based portal that allows users to submit their ontologies, to associate metadata with their ontologies, to search for existing ontologies, to find ontology rankings based on user reviews, to post their own reviews, and to rate reviews. PMID- 17911830 TI - PICO Linguist and BabelMeSH: development and partial evaluation of evidence-based multilanguage search tools for MEDLINE/PubMed. AB - PICO Linguist and BabelMeSH are multilanguage search tools intended for users whose native language is not English. A database of medical terms was created using concept identification equivalents of English terms to other languages. The primary sources of vocabularies were UMLS, MeSH, WHO EMRO and UMLF. The search interface changes according to the language selected which allows search terms to be entered in the native language. The user can limit the search output according to the language of publication but citations retrieved are in English only. Links may be provided to journals if published online. Evaluation of the French and Spanish versions using journal key words and a list of common diseases showed 77.5% and 86.5% accuracy respectively. User feedback was positive. PICO Linguist and BabelMeSH could be useful and convenient tools in finding current evidence sources in the medical literature especially for non-English medical terms that may be difficult to express in English. PMID- 17911831 TI - Lessons learned from cross-validating alignments between large anatomical ontologies. AB - OBJECTIVES: To compare the alignments of two large anatomical ontologies (the Foundational Model of Anatomy and GALEN) produced by three ontology alignment systems (AOAS, FALCON and PRIOR) in the framework of the Ontology Alignment Evaluation Initiative during its 2006 campaign. MATERIALS: Number of mappings identified by AOAS: 3,132, FALCON: 2,518 and PRIOR: 2,589. METHODS: Three approaches to analyzing and comparing the results were utilized: computing the overlap among result files, manual review of some 2,000 mappings and structural validation. CONCLUSIONS: The generic systems FALCON and PRIOR identify many false positives and false negatives. With a stricter and domain-specific lexical similarity model, AOAS has a better precision, but is more sensitive to missing synonyms and misspellings. PMID- 17911832 TI - How updating textual clinical practice guidelines impacts clinical decision support systems: a case study with bladder cancer management. AB - Guideline-based clinical decision support systems (CDSSs) can be effective in increasing physician compliance with recommendations. However, the ever growing pace at which medical knowledge is produced requires that clinical practice guidelines (CPGs) be updated regularly. It is therefore mandatory that CDSSs be revised accordingly. The French Association for Urology publishes CPGs on bladder cancer management every 2 years. We studied the impact of the 2004 revision of these guidelines, with respect to the 2002 version with a CDSS, UroDoc. We proposed a typology of knowledge base modifications resulting from the update of CPGs making the difference between practice, clinical conditions and recommendations refinement as opposed to new practice and new recommendations. The number of formalized recommendations increased from 577 in 2002 to 1,081 in 2004. We evaluated the two versions of UroDoc on a randomized sample of patient records. A single new practice that modifies a decision taken in 49% of all recorded decisions leads to a fall from 67% to 46% of the compliance rate of decisions. PMID- 17911833 TI - Improving compliance to guidelines through workflow technology: implementation and results in a stroke unit. AB - This work describes the results of the implementation of a workflow management system integrated into the electronic clinical chart of a Stroke Unit. The workflow logic is based on the rules provided by the SPREAD guidelines for stroke management. In this way, the already existing clinical chart has been transformed into an evidence-based, real-time decision support system, meanwhile maintaining the same look the users were familiar with. Since the final aim of the work was to improve evidence-based behavior and detect possible organizational bottlenecks, non-compliance to the clinical practice guidelines, before and after the system introduction, have been analyzed, as well as the accuracy of the clinical chart compilation, some care process variables, and system usability. Results show that the system enhances the clinical practice without boring users. Moreover, non-compliance analysis gives rise to ideas for further improvement. PMID- 17911834 TI - Towards a decision support system for optimising clinical pathways of elderly patients in an emergency department. AB - Data stored in Healthcare Information Systems correspond potentially to a vast source of information for supporting decisions in management or public health issues. The presented study illustrates clinical data valuation, through the analysis of clinical pathways of elderly patients at the Emergency Department (ED) of Rennes hospital. METHOD: Relevant data were extracted from the Emergency Department database. Several analysis (e.g., cusum method) and representation tools (e.g., Graphviz) were used to study the patients' pathways, the dynamics of flows and the patients' characteristics. RESULTS: 4951 admissions were analysed and visualized. The representations provided a synthetic, global and comprehensive view of the department activities, to the satisfaction of the clinicians. Limitations of the ICD-10 coding of the diagnoses at the ED were pointed out as well as syntax and semantic interoperability issues. A solution is proposed for automating and scaling the Decision Support System. PMID- 17911835 TI - Ontology-based modeling of clinical practice guidelines: a clinical decision support system for breast cancer follow-up interventions at primary care settings. AB - Breast cancer follow-up care can be provided by family physicians after specialists complete the primary treatment. Cancer Care Nova Scotia has developed a breast cancer follow-up Clinical Practice Guideline (CPG) targeting family physicians. In this paper we present a project to computerize and deploy the said CPG in a Breast Cancer Follow-up Decision Support System (BCF-DSS) for use by family physicians in a primary care setting. We present a semantic web approach to model the CPG knowledge and employ a logic-based proof engine to execute the CPG in order to infer patient-specific recommendations. We present the three stages of the development of BCF-DSS--i.e., (a) Computerization of the paper based CPG for Breast Cancer follow-up; (b) Development of three ontologies--i.e., the Breast Cancer Ontology, the CPG ontology based on the Guideline Element Model (GEM) and a Patient Ontology; and (c) Execution of the Breast Cancer follow-up CPG through a logic-based CPG execution engine. PMID- 17911836 TI - Learning causal and predictive clinical practice guidelines from data. AB - Clinical practice guidelines (CPG) propose preventive, diagnostic and treatment strategies based on the best available evidence. CPG enable practice of evidencebased medicine and bring about standardization of healthcare delivery in a given hospital, region, country or the whole world. This study explores generation of guidelines from data using machine learning, causal discovery methods and the domain of high blood pressure as an example. PMID- 17911837 TI - Supporting therapy selection in computerized clinical guidelines by means of decision theory. AB - Supporting therapy selection is a fundamental task for a system for the computerized management of clinical guidelines (GL). The goal is particularly critical when no alternative is really better than the others, from a strictly clinical viewpoint. In these cases, decision theory appears to be a very suitable means to provide advice. In this paper, we describe how algorithms for calculating utility, and for evaluating the optimal policy, can be exploited to fit the GL management context. PMID- 17911838 TI - Development, deployment and usability of a point-of-care decision support system for chronic disease management using the recently-approved HL7 decision support service standard. AB - Clinical decision support is recognized as one potential remedy for the growing crisis in healthcare quality in the United States and other industrialized nations. While decision support systems have been shown to improve care quality and reduce errors, these systems are not widely available. This lack of availability arises in part because most decision support systems are not portable or scalable. The Health Level 7 international standard development organization recently adopted a draft standard known as the Decision Support Service standard to facilitate the implementation of clinical decision support systems using software services. In this paper, we report the first implementation of a clinical decision support system using this new standard. This system provides point-of-care chronic disease management for diabetes and other conditions and is deployed throughout a large regional health system. We also report process measures and usability data concerning the system. Use of the Decision Support Service standard provides a portable and scalable approach to clinical decision support that could facilitate the more extensive use of decision support systems. PMID- 17911839 TI - The TAR model: use of therapeutic state transitions for quality assurance reporting in chronic disease management. AB - Chronic disease management represents one of the challenges for health informatics and demands the appropriate application of information technology for improved patient care. This paper presents an approach to quality assurance reporting wherein the recommendations of evidence-based clinical practice guidelines are considered in the context of empirical therapeutic state transitions (in terms of changes in individual patient prescriptions over time). We apply a Transition-based Audit Report (TAR) model to antihypertensive prescribing and related data as stored in a New Zealand General Practice Management System database. The results provide a set of quality indicators and specific patient cohorts for potential practice quality improvement with strong linkage to the selected guidelines and observed practice patterns. We see the TAR model primarily as a tool to enable internal quality improvement efforts, but also to be of relevance for focusing pay-for-performance programs. PMID- 17911840 TI - Development of case-based medication alerting and recommender system: a new approach to prevention for medication error. AB - The purpose of this study was to develop a new alerting and recommender system for preventing medication errors. In recent years, alerting systems have been widely implemented, but because these systems apply a same static threshold for all patients in all cases, they produce excessive alerts and subject physicians to "alert fatigue". We believe that the most commonly-written prescription for a patient's status is the safest one. From this standpoint, we developed a real time case-based medication alerting and recommender system linked to a database of past prescriptions. When a physician issues his or her prescription, our system dynamically compares it with past ones for similar patients in the database. An analysis of the 10 most frequently-used drugs in the University of Tokyo Hospital revealed that our system reduced the number of false alerts compared to the traditional static alert method. Our system contributes to the creation of alerts that are appropriate for patients' clinical conditions and based on physicians' empirical discretion. PMID- 17911841 TI - Is the future evidence-based? AB - This paper is concerned with how the future information needs of the medical community should be met. The current dominant belief within medicine is that these information needs should be met from bespoke research studies. The necessity of this approach is far from certain. Health organisations worldwide are currently investing vast resources into centralising and amalgamating every day patient data. Is there a future for these Electronic Medical Records in informing medical decisions? This paper describes the challenges to be met in using both research studies and every day patient data to inform medical decisions. It then describes an ongoing practical project to evaluate these information sources' ability to meet the information needs of cancer care decision makers. Details of background, methodology and initial promising results are presented. PMID- 17911842 TI - Comparing decision support methodologies for identifying asthma exacerbations. AB - OBJECTIVE: To apply and compare common machine learning techniques with an expert built Bayesian Network to determine eligibility for asthma guidelines in pediatric emergency department patients. POPULATION: All patients 2-18 years of age presenting to a pediatric emergency department during a 2-month study period. METHODS: We created an artificial neural network, a support vector machine, a Gaussian process, and a learned Bayesian network to compare each method's ability to detect patients eligible for asthma guidelines. Our outcome measures included the area under the receiver operating characteristic curves, sensitivity, specificity, predictive values, and likelihood ratios. RESULTS: The data were randomly split into a training set (n=3017) and test set (n=1006) for analysis. The systems performed equally well. The area under the receiver operating characteristic curve was 0.959 for the expert-built Bayesian network, 0.962 for the automatically constructed Bayesian network, 0.956 for the Gaussian Process, and 0.937 for the artificial neural network. DISCUSSION: All four evaluated machine learning methods achieved high accuracy. The automatically created Bayesian network performed similarly to the expert-built network. These methods could be applied to create a realtime detection system for identifying asthma patients. PMID- 17911843 TI - Analysis and redesign of a knowledge database for a drug-drug interactions alert system. AB - Physicians tend to ignore drug-drug interactions alerts, this is due to the large amount of irrelevant interactions displayed and the interface in which these alerts are shown. The high rate of clinically inadequate alerts produce "alerts fatigue". This high number of incorrect alerts predisposes physicians to underestimate the electronic prescription systems as useful tools in their practice. We decided to analyze and redesign our drug-drug interactions alerting system knowledge database. In order to do so, we cleaned our knowledge database according to the clinical significance of drug-drug interactions. New drug interactions taxonomy was created in four levels based on clinical significance and the recommendations given in each single monograph of interaction. We proceeded to recategorize the alerts as Active, which present themselves to the physician interrupting the prescribing process, or Passive, which allow physicians to accept the recommendations, and adopt some action in order of minimizing the interaction risks. PMID- 17911844 TI - Closing the loop: bringing decision support clinical data at the clinician desktop. AB - We describe the development of an inquiry office to bridge the gap between clinician needs for decision support systems and readily available large quantities of integrated clinical data. With this link, an information feedback mechanism is implemented that closes the loop of information flow by bringing decision support information from the data warehouse at the clinician desktop. As a result, and as a new DRG cost reimbursement system has been introduced, we have provided the heads of over 30 medical services with an intranet web-based application to access patient encoding of diagnoses, procedures, and DRGs of their respective service. The inquiry office has also developed a query service to process specific requests. It has implemented the automatic screening of patient clinical data of past and current hospitalizations in order to select cases for multiple studies, research, and teaching projects. The purpose of this clinical data warehouse and its information feedback process is to offer a coherent, comprehensive, and reliable return of information to improve decision making, to enable research projects, and to facilitate statistical outputs. PMID- 17911845 TI - Are clinicians' information needs and decision support affected by different models of care? Experimental study. AB - This study explores task- and healthcare model-specific differences in clinicians' information needs which can affect the uptake of decision support. Results of a web experiment involving 104 general practitioners are presented. Respondents indicated that guidelines were the most important source of information with almost equal weighting for acute, chronic and preventive care. A patient's quality of life was identified as the most important determinant of decision-making in all three models of care. Risk assessment tools and information about outcomes were more valuable (P<0.05) for chronic and preventive care than for acute cases. The participants accessed electronic risk assessment tools in 54%, 45% and 81% of acute, chronic and preventive care scenarios, respectively. Participants estimated that the electronic decision support would have a significantly higher impact in preventive care than in chronic or acute care settings (P=0.01). The differences in the information needs of clinicians related to different care models have to be considered in the design of clinical decision support systems. Systems that target preventive model decisions may have higher adoption and impact. PMID- 17911846 TI - Modeling and acquisition of drug-drug interaction knowledge. AB - OBJECTIVES: The effectiveness of computerized clinical decision support systems (CDSS) depends on the quality of the knowledge they refer to. In this article, we are interested in the acquisition, modeling and representation of the knowledge embedded in the "national reference framework of drug-drug interaction" published by the French Health Products Safety Agency. METHODS: A model of drug-drug interactions has been designed using bottom-up and top-down approaches. This model is the basis for the design of an XML format to represent and extract information on drug interactions from the reference framework. RESULTS: A specific tool has been developed to extract the information from a corpus of 1053 drug monographs using a methodology similar to the one used by the GEM-Cutter tool to extract information from clinical guidelines. Strategies to integrate the XML files produced into CDDSSs are discussed. DISCUSSION-CONCLUSION: Modeling and acquisition of drug-drug interaction knowledge from a corpus of drug monographs is a potential approach to foster the development of CDSS and improve their specificity. PMID- 17911847 TI - A systems development life cycle approach to patient journey modeling projects. AB - Patient Journey Modeling, a relatively recent innovation in healthcare quality improvement, models the patient's movement through a Health Care Organisation (HCO) by viewing it from a patient centric perspective. A Systems Development Life Cycle (SDLC) provides a standard project management framework that can improve the quality of information systems. The concept of following a consistent project management framework to boost quality outcomes can be applied equally to healthcare improvement. This paper describes a SDLC designed specifically for the health care domain and in particular patient journey modeling projects. It goes on to suggest that such a framework can be used to compliment the dominant healthcare improvement method, the Model for Improvement. The key contribution of this paper is the introduction of a project management framework in the form of an SDLC that can be used by non-professional computer developers (ie: health care staff), to improve the consistency and quality of outcomes for patient journey redesign projects. Experiences of applying the SDLC in a midwife-led primary-care maternity services environment are discussed. The project team found the steps logical and easy to follow and produced demonstrable improvement results along with ongoing goal-focused action plans. PMID- 17911848 TI - The nurse-patient trajectory framework. AB - The development of nursing knowledge should give structure and form to the practice of nursing. The development of Nursing Process Theory resulted from early nursing observations and inferences from nursing practice that resulted in formal data accumulation processes, mutual correspondence between nurses and patients, and exchange of information. The development of the nursing process discipline helped to substantiate the need for professional nursing services. The shifts towards examining the links between processes and outcomes, professional accountability, and classification of distinct nursing functions have influenced the development of information systems. The Nurse-Patient Trajectory Framework described in this paper may be used to show the relationships between the virtual information system and the real world that it affects. The framework is visualized along two separate and distinct nurse and patient trajectories. PMID- 17911849 TI - System analysis and improvement in the process of transplant patient care. AB - Clinical information concerning transplant patients is voluminous and difficult to manage using paper records. A system analysis was performed to assess information system needs of the liver, kidney, and pancreas transplant program at LDS Hospital in Salt Lake City, Utah. After evaluating workflow, decision support needs, and requirements, we designed and implemented an extendable information system to support care following liver transplantation. We developed and implemented a standardized operative note, forms to enter external laboratory results and transplant-related information into the electronic health record, and computerized alerts to notify the transplant nurses when liver transplant patients had new, abnormal, or overdue laboratory results. The information system has improved the quality of clinical data available in the EHR, clinician satisfaction, and efficiency with management of laboratory results. The components developed for this project can be extended to meet other transplant program needs. PMID- 17911850 TI - St Elsewhere's or St Everywhere's: improving patient throughput in the private hospital sector. AB - Communication errors have been found to be most common root cause of medical errors by the US-based Agency for Healthcare Research and Quality [1]. Although elective admissions to hospital involves a high volume of important healthcare communications where incorrect, missing or illegible information could result in a serious medical error, there is little published research on the impact of improving pre-admission communication flow between admitting doctors and hospitals. The Sydney Adventist Hospital (the San) is a 341-bed private hospital in Sydney's northern suburbs that provides a comprehensive range of health services. A process improvement program began in early 2005 to streamline preadmission communications. The objectives of this ongoing program are broadly to improve patient safety and to increase operating efficiency. The first major initiative within this program was to implement a standardised method for inpatient booking/referral with over three hundred admitting doctors. Eighteen months on, the hospital has been able to demonstrate a significant shift in the timeliness of patient bookings from specialists' rooms, more comprehensive provision of clinical indicators that can facilitate resource planning in operating theatres and on the wards, and reduction in the ratio of bookings made in areas other than the hospital bookings department. The program continues with focus on improving accuracy of data entry, rationalising patient forms, making more effective use of information received and automation of pre-admission information flows. PMID- 17911851 TI - A meta schema for evidence information in clinical practice guidelines as a basis for decision-making. AB - Clinical practice guidelines are an important instrument to aid physicians during medical diagnosis and treatment. Currently, different guideline developing organizations try to define and integrate evidence information into such guidelines. However, the coding schemas and taxonomies used for the evidence information differ widely, which makes the use cumbersome and demanding. We explored these various schemas and developed a meta schema for the evidence information, which covers the most important components of the existing ones, is comprehensible, and easy to understand for the users. We developed and assessed the usefulness and applicability of our meta schema with guideline developers and physicians. PMID- 17911852 TI - Creating interoperable guidelines: requirements of vocabulary standards in immunization decision support. AB - Interoperable support of electronic health records and clinical decision support technology are central to the vision of sustainable information infrastructure. Efforts to implement interoperable clinical guidelines for immunization practice have been sparse. We used the SAGE knowledge workbench to develop a knowledge base to provide immunization decision support in primary care. We translated the written clinical guideline into a structured decision logic format. The semantic content to completely capture CDC clinical decision logic required 197 separate concepts but was completely captured with SNOMED CT and LOINC. Although 88% of concepts employed precoordinated codes, 6% of guideline concepts required expanded vocabulary services employing Boolean logical definition using two or more SNOMED concepts. Postcoordination requirements were modest, representing just 6% of guideline semantic concepts. We conclude that creation of interoperable knowledge bases employing clinical vocabulary standards is achievable and realistic. Employment of information model (HL7 RIM) and vocabulary (SNOMED CT, LOINC) standards is a necessary and feasible requirement to achieve interoperability in clinical decision support. PMID- 17911853 TI - Automatic treatment of temporal issues in clinical guidelines in the GLARE system. AB - Temporal constraints play a fundamental role in clinical guidelines. For example, temporal indeterminacy, constraints about duration, delays between actions and periodic repetitions of actions are essential in order to cope with clinical therapies. This paper proposes a computer-based approach in order to deal with temporal constraints in clinical guidelines. Specifically, it provides the possibility to represent such constraints and reason with them (i.e., perform inferences in the form of constraint propagation). We first propose a temporal representation formalism and two constraint propagation algorithms operating on it, and then we show how they can be exploited in order to provide clinical guideline systems with different temporal facilities. Our approach offers several advantages: for example, during the guideline acquisition phase, it enables to represent temporal constraints and to check their consistency; during the execution phase, it allows the physician to check the consistency between action execution-times and the constraints in the guidelines, and to provide query answering and temporal simulation facilities (e.g., when choosing among alternative paths in a guideline). PMID- 17911854 TI - Information and communication processes in the microbiology laboratory- implications for computerised provider order entry. AB - The aim of this multi-method study based at a microbiology department in a major Sydney metropolitan teaching hospital was to: i) identify the role that information and communication processes play in a paper-based test request system, and ii) examine how these processes may affect the implementation and design of Computerised Provider Order Entry (CPOE) systems. Participants in this study reported that clinical information can impact on the urgency and type of tests undertaken and affect the interpretation of test results. An audit of 1051 microbiology test request forms collected over a three-day period showed that 47% of request forms included clinical notes which provide a variety of information often vital to the test analysis and reporting process. This transfer of information plays an important role in the communication relationship between the ward and the laboratory. The introduction of new CPOE systems can help to increase the efficiency of this process but for that to be achieved research attention needs to be given to enhancing the provision and communication of clinical information. PMID- 17911855 TI - Using an accident model to design safe electronic medication management systems. AB - Large-scale implementation of electronic prescribing systems (e-PS) is likely to introduce at least some machinerelated errors that will harm patients. We present a dynamic systems modeling approach to developing a comprehensive multilevel accident model of the process, context and task interaction variables which give rise to human error and system failure when e-PS are used in routine care. System dynamics methods are used to represent interactions between medication management processes and the context that is relevant to error generation, interception and transmission, agent-based methods represent task interactions. Capturing the patterns of failure within an accident model will facilitate an evidence-based approach to hazard analysis and design of e-PS features to improve patient safety. The model will have broad potential to guide the design, implementation and regulation of e-PS. PMID- 17911856 TI - Securing chemotherapies: fabrication, prescription, administration and complete traceability. AB - Decision support, order entry, drug and care administration with their respective documentation cannot be seen as independent actions, especially in term of medical approach and patient safety. Chemotherapy treatment is a good example to illustrate the various implication of technology information in these multifaceted and intricate processes. Chemotherapy administration can be a highly complex process. It can take place over a variable period of time, from hours to months. Chemotherapies can be produced specifically for a given patient and can have dramatic effects in case of errors. Chemotherapy treatment will depend from various information including patient specific data such as temperature, weight or laboratory or drug specific knowledge such as side effects or administration directives. At the University Hospitals of Geneva (HUG), processes reengineering accompanied with new applications covering the whole chain of the processes involved in chemotherapy treatment (prescription preparation, administration, control) have been developed. This paper presents the overall approach leading the computerization of these processes and the first evaluations about the potential benefits of the computer-aided controls during the administration phase. PMID- 17911858 TI - Diffusion of electronic health records--six years of empirical data. AB - The Danish EHR-Observatory has monitored Danish EHR projects since 2000 with respect to a number of parameters such as diffusion, diffusion rate and the hospital owners expectations. On the basis of the data and a model for technology diffusion three scenarios for future diffusion are built. The results show that the national goal to have EHRs fully implemented in hospitals by 2008 will not be reached. The scenarios built from empirical data provide a useful benchmarking tool for planning and evaluating the EHR implementation programs in hospitals. PMID- 17911857 TI - Multitasking by clinicians in the context of CPOE and CIS use. AB - Interest in studying distractions and interruptions in the context of clinician workflow has increased in light of evidence that these events may negatively impact patient safety. Additionally, many recent informatics-based studies analyze computer provider order entry (CPOE) and clinical information system (CIS) implementation and its effects on clinician workflow. This study expands the development and use of a taxonomy to characterize distractions to clinicians and their subsequent actions in the context of CPOE/CIS use. We found a total of 75 distracting events in 406 minutes of observational data of which 32 led to interruptions and 30 led to continued multitasking. The above primary actions led to 5 tasks not completed and 4 episodes of clinician's lack of recall. Differences in the distribution of the source of distractions and primary action of the distracted clinicians may be a function of clinical setting and clinician type using the CPOE/CIS. Nine secondary actions, potentially resulting in a slip or a mistake, suggest that CPOE may necessitate different forms of safety nets than traditional clinician communication. PMID- 17911859 TI - Text categorization models for identifying unproven cancer treatments on the web. AB - The nature of the internet as a non-peer-reviewed (and largely unregulated) publication medium has allowed wide-spread promotion of inaccurate and unproven medical claims in unprecedented scale. Patients with conditions that are not currently fully treatable are particularly susceptible to unproven and dangerous promises about miracle treatments. In extreme cases, fatal adverse outcomes have been documented. Most commonly, the cost is financial, psychological, and delayed application of imperfect but proven scientific modalities. To help protect patients, who may be desperately ill and thus prone to exploitation, we explored the use of machine learning techniques to identify web pages that make unproven claims. This feasibility study shows that the resulting models can identify web pages that make unproven claims in a fully automatic manner, and substantially better than previous web tools and state-of-the-art search engine technology. PMID- 17911860 TI - Currency of online breast cancer information. AB - Consumers are increasingly turning to the Web, expecting to find the latest health information. The purpose of this study was to assess the currency of online breast cancer information. We determined whether nine recent advances in breast cancer management were incorporated into 337 unique breast cancer Web pages. Two reviewers independently assessed content; if a Web page covered appropriate advances it was deemed to be "current." Of the 337 Web pages, 89 contained one or more advances. Of the 122 Web pages that had dates of update available, 49% had been updated within 6 months. Only 11%-37% of Web pages covered clinically accepted advances, even among Web pages that were updated after acceptance of the advance into clinical practice. We conclude that online health information is often not sufficiently current. Consumers searching for health information online should always consult an expert clinician before taking action. PMID- 17911861 TI - Children's contributions to designing a communication tool for children with cancer. AB - In this paper, we describe the roles played as well as contributions made by child participants in the design of an innovative communication tool for children with cancer. SISOM is a handheld, portable computer application with a graphical user interface that is meant to: (1) help children with cancer communicate their symptoms / problems in a child-friendly, age-adjusted manner; and (2) assist clinicians in addressing children's experienced symptoms and problems in patient care. Unlike other applications for children, the purpose of SISOM is not to provide information to ill children but to elicit personal information from them. Thus the application has a unique set of design issues. Healthy and ill children played an important role in different stages in the design process. They made significant contributions to the graphical design of the system's interface; selection of understandable, child-friendly terms used by the system to describe symptoms; iconic and graphical representations; and its usability. We describe the participatory design methods we used that included children and share important insights from this collaborative design process. PMID- 17911862 TI - "It's your game": an innovative multimedia virtual world to prevent HIV/STI and pregnancy in middle school youth. AB - Early sexual initiation is associated with increased risk of unintended pregnancy and sexually transmitted infection(STI). Effective HIV/STI/pregnancy prevention interventions for middle school youth are urgently needed. "It's Your Game, Keep It Real" (IYG) is a curriculum delivered in 7th and 8th grade that combines classroom activities with individualized, tailored computer-based activities embedded in a 'virtual world' environment. Interactive multimedia can offer a confidential, tailored, and motivational educational experience. Virtual world game interfaces offer further potential to immerse the learner. The purpose of this study was to evaluate the multimedia education program component of IYG on student attitudes of importance of the curriculum content, self-efficacy regarding refusal skills, and usability parameters of ease of use, credibility, understandability, acceptability, and motivation to determine that a broader efficacy field test would be indicated. Results of the study indicated acceptable usability criteria and impact on short-term psychosocial outcomes. IYG is currently being evaluated in a randomized controlled trial in ten Texas middle schools. PMID- 17911863 TI - HeartCareII: home care support for patients with chronic cardiac disease. AB - Systematic engagement of patients in disease management requires design and deployment of innovative technologies that complement and extend professional nursing services. We describe here a model of nursing practice that capitalizes on a web-based resource (HeartCareII) to support patient self-management, symptom interpretation, and self-monitoring. Research staff provided computers and technical assistance; visit nurses trained patients in the components of the HeartCareII Website most relevant to their care needs. This paper describes the nursing practice model and the web resource, and reports the experience of patients recruited in the early phase of the study. PMID- 17911864 TI - A web-based communities of practice support system for caregivers. AB - CareNet is an interactive Web-based system intended to support informal caregivers (ordinary citizens who are engaged in providing residential healthcare to their families and friends). The design of CareNet uses concepts from a number of areas including: Communities of Practice, software engineering, content authoring, and knowledge management to create a supportive environment for the caregiver. The specific objectives of the CareNet project are: (1) To create a highly effective interactive environment that addresses the needs of caregivers to: (1a) obtain information and guidance, (1b) achieve efficient communication with professional healthcare workers with whom they collaborate and with other caregivers from whom they can gain advice, emotional support and solace, (1c) access other information and physical resources that are needed for proper care, and (1d) document their observations, interventions and insights that can in future become a knowledge resource for other caregivers; and (2) To demonstrate the beneficial impact of CareNet on caregivers who collaborate with professional care providers and on their interactions with professional care providers. PMID- 17911865 TI - Core features of a parent-controlled pediatric medical home record. AB - We describe a coordinated effort to identify the core features of a parent controlled personal health record for children with special health care needs, involving parents, care givers, and healthcare providers. A summary of the core features is presented, emphasizing needs that are not commonly recognized as functions of a generic personal health record. Our goal was to identify requirements for personal records that empower parents to effectively obtain, organize, understand, and communicate the information necessary to help their children receive the best possible care. PMID- 17911867 TI - Clinical communication ontology for medical errors. AB - Clinical communication failures caused 60% of sentinel events reported by the Joint Commission on Accreditation of Healthcare Organizations. The difficulties of communication have been the primary cause of errors leading to patients' death. For analyzing medical error events, uncovering the patterns of clinical communication, this paper reports the design and development of clinical communication ontology. The ontology contains eight axes and was validated using ten medical error cases, where communication was the main factor. The coding process demonstrates that the ontology can be used as a guideline for future medical error reporting system, through which the root cause of medical error due to communication will be revealed in a clear pattern. This ontology contributes to the generation of proper interventions and effective strategies for reducing medical errors. PMID- 17911866 TI - Empowering patients to improve the quality of their care: design and implementation of a shared health maintenance module in a US integrated healthcare delivery network. AB - We describe a health maintenance module within a personal health record designed to improve the quality of routine preventive care for patients in a large integrated healthcare delivery network. This module allows patients and their providers to share an online medical record and decision support tools. Our preliminary results indicate that this approach is well-accepted by patients and their providers and has significant potential to facilitate patient-provider communication and improve the quality of routine health maintenance care. Further research will determine the long term impact and sustainability of this approach. PMID- 17911868 TI - u-SHARE: web-based decision support/risk communication tool for healthcare consumers with unruptured intracranial aneurysms. AB - PURPOSE: Clinical management for unruptured intracranial aneurysms (UIA) is controversial and requires professional knowledge which is the main reason that patients have difficulty in making decisions. The purpose of this study is to develop a tool that aids healthcare consumers in making optimal shared decisions with decision analysis. METHODS: The decision model and relevant data were derived from published literature. A web-based decision analytic tool was designed to provide a systematic guide for patients to understand favorable treatment options, intrinsic uncertainty, and critical factors for decision making. Twenty-nine testers evaluated content appropriateness, usability and clinical usefulness of the tool. RESULTS: The decision analytic tool has been successfully implemented and evaluated. Testers generally judged the web-based decision analytic tool as functional and useful. Acceptance rate for decision analysis was higher in nonhealthcare professionals than health care professionals. CONCLUSIONS: Our decision analytic tool was well accepted especially by healthcare consumers. The tool enables UIA patients to enhance their knowledge and understanding toward optimal shared decision making and can be an alternative "structured informed consent tool". PMID- 17911870 TI - The use of electronic medication reconciliation to establish the predictors of validity of computerized medication records. AB - Medication records in clinical information systems (CIS) are frequently inaccurate, leading to potentially incorrect clinical decisions and preventing valid decision support interventions. It is not known what characteristics of electronic medication records are predictive of their validity. We studied a dataset of 136,351 electronic medication records of patients admitted to two academic hospitals that were individually validated by admitting providers using novel medication reconciliation software. We analyzed the relationship between characteristics of individual medication records and the probability of record validation using a multivariable linear regression model. Electronic medication records were less likely to be validated if more time had passed since their last update (14.6% for every 6 months), if they represented an antiinfective (61.6%) or a prn (50.9%) medication, or if they were in an outpatient CIS rather than on an inpatient discharge medication list (18.1%); p<0.0001 for all. Several characteristics of electronic medication records are strongly associated with their validity. These findings could be incorporated in the design of CIS software to alert providers to medication records less likely to be accurate. PMID- 17911869 TI - Health on the Net Foundation: assessing the quality of health web pages all over the world. AB - The Internet provides a great amount of information and has become one of the communication media which is most widely used [1]. However, the problem is no longer finding information but assessing the credibility of the publishers as well as the relevance and accuracy of the documents retrieved from the web. This problem is particularly relevant in the medical area which has a direct impact on the well-being of citizens. In this paper, we assume that the quality of web pages can be controlled, even when a huge amount of documents has to be reviewed. But this must be supported by both specific automatic tools and human expertise. In this context, we present various initiatives of the Health on the Net Foundation informing the citizens about the reliability of the medical content on the web. PMID- 17911871 TI - Evaluation of an electronic medication reconciliation system in inpatient setting in an acute care hospital. AB - BACKGROUND: Medication reconciliation (MedRecon) is being implemented in many healthcare facilities as a means to reduce medication errors. However, there is scant literature on the evaluation of electronic MedRecon systems. OBJECTIVE: To evaluate the rate and type of discrepancies between a patient's home medication history and admission orders and to analyze factors affecting their occurrence using an electronic MedRecon system. DESIGN/METHODS: We analyzed 3,426 consecutive inpatient admission MedRecon events from August to October 2006 in an acute care hospital using a recently implemented electronic MedRecon system. RESULTS: Overall, discrepancy rate was 3.12% (n=107) with omission of a home medication being the most common type (56.52%, n=65) of discrepancy. Admission time(8 PM to 8 AM), and total home medications>4 were found to have a significant positive correlation with discrepancy rate. CONCLUSION: Using multidisciplinary MedRecon process based on an electronic system, we found a low discrepancy rate between patient's home medication history and admission orders compared with the rate in the literature, implying that an electronic MedRecon system is an important tool for improving patient safety. PMID- 17911872 TI - Computerized management of chronic anticoagulation: three years of experience. AB - Chronically anticoagulated patients taking the drug Warfarin require time intensive management and followup processes to avoid complications. The "Chronic Anticoagulation Clinic" (CAC) protocol is a set of production rules that help manage, treat, and follow-up such patients. The CAC protocol has been in regular use at Intermountain Healthcare (Salt Lake City, UT, USA) for over three years. The results demonstrate an improvement on the number of patients with anticoagulation levels within the desired target range. The protocol alerts have a high acceptance rate (83.4%) and were able to help patients remember to collect their next coagulation test. The CAC protocol results show that production rules can improve the management of chronically anticoagulated patients. Additional studies are required to verify if this experience can be transferred to other institutions. PMID- 17911873 TI - Physicians' response to guided geriatric dosing: initial results from a randomized trial. AB - Guided dosing within a computerized provider order entry (CPOE) system is an effective method of individualizing therapy for patients. Physicians' responses to guided dosing decision support have not been extensively studied. As part of a randomized trial evaluating efficacy of dosing advice on reducing falls in the elderly, CPOE prompts to physicians for 88 drugs included tailored messages and guided dose lists with recommended initial doses and frequencies. The study captured all prescribing activity electronically. The primary outcome was the ratio between prescribed dose and recommended dose. Over 9 months, 778 providers entered 9111 study-related medication orders on 2981 patients. Physicians using guided orders chose recommended doses more often than controls(28.6% vs. 24.1%, p<0.001). Selected doses were significantly lower in the intervention group (median ratio of actual to recommended 2.5, interquartile range [1.0,4.0]) than the control group (median 3.0 interquartile range [1.5,5.0], p<0.001). While physicians selected the recommended dose less than a third of the time, guided geriatric dosing modestly improved compliance with guidelines. PMID- 17911874 TI - Graphical overview and navigation of electronic health records in a prototyping environment using Google Earth and openEHR archetypes. AB - This paper describes selected earlier approaches to graphically relating events to each other and to time; some new combinations are also suggested. These are then combined into a unified prototyping environment for visualization and navigation of electronic health records. Google Earth (GE) is used for handling display and interaction of clinical information stored using openEHR data structures and 'archetypes'. The strength of the approach comes from GE's sophisticated handling of detail levels, from coarse overviews to fine-grained details that has been combined with linear, polar and region-based views of clinical events related to time. The system should be easy to learn since all the visualization styles can use the same navigation. The structured and multifaceted approach to handling time that is possible with archetyped openEHR data lends itself well to visualizing and integration with openEHR components is provided in the environment. PMID- 17911875 TI - Which parts of a clinical process EPR needs special configuration. AB - SUBJECT: Which parts of an electronic patient record (EPR) can initially form a stable standard solution to be used by all clinicians? And which parts of an EPR can we predict needs initial as well as on-going re-configuration to meet the needs from diverse medical specialties. PURPOSE: To analyze which screen types in a clinical process that can be standard configured and which are subject to initial as well as on-going re-configuration. METHODS AND RESULTS: A pilot project implementing a fully functional clinical process EPR was configured and used at a neurological ward, replacing all paper records 24/7. The analysis characterizes the different types of screens, a total of 243 included in the EPR solution. All screens have been extracted from the application and analyzed for changes in total 222 changes. DISCUSSION AND CONCLUSION: Most screens (87%) are very stable. Few (13%) are subjected to several re-configurations and they stabilize after an average of six iterations: Some may further stabilize over time since they address new but also general ways of working. Other screens relate to the specific medical specialty and cannot be part of a standard solution. PMID- 17911876 TI - User driven, evidence based experimental design; a new method for interface design used to develop an interface for clinical overview of patient records. AB - A novel method of interface design--user driven, evidence based experimental design--was developed which approximates the usual clinical way of maturing science and technology in the healthcare domain. The method is user-driven and the clinician remains in control of gathering and evaluating evidence of relevance to the project--as well as specifying the details of the user interface. Information not obtainable from the literature was gained experimentally and used to achieve a deeper understanding of the problem before the design phase. The design was subsequently validated experimentally by ordinary users with no connection to the software or design team. After applying this method to the problem of gaining a satisfactory clinical overview of a single patient's record, we recommend that clinical IT interfaces have clinical logic, sufficient complexity, and are well structured. Developers should use computer power to support "building blocks" such as anatomical problem lists and summaries of history, status and treatment, personal notes, and should support clinical browsing using text and graphics. PMID- 17911877 TI - User interface optimization for an electronic medical record system. AB - Many information technology-enabled healthcare applications have failed because their interfaces are difficult to use. Unfortunately, little attention has been paid in the health informatics community to designing effective user interfaces that are acceptable to healthcare professionals. This paper illustrates a method for improving application interface usability by applying sequential pattern analysis to analyze temporal event sequences recorded in an electronic medical record system. Such event sequences, or clickstreams, reflect clinicians' navigation patterns in their everyday interactions with the computer system. The identified patterns have been used by software developers to calibrate the user interface of the system, so that the within-application workflow is better aligned with clinicians' mental model of medical problem-solving. Such inferred patterns may also help to modify clinicians' suboptimal practice behavior components, as manifested through their actual usage of this point-of-care electronic system. PMID- 17911878 TI - AdaRTE: adaptable dialogue architecture and runtime engine. A new architecture for health-care dialogue systems. AB - Spoken dialogue systems have been increasingly employed to provide ubiquitous automated access via telephone to information and services for the non-Internet connected public. In the health care context, dialogue systems have been successfully applied. Nevertheless, speech-based technology is not easy to implement because it requires a considerable development investment. The advent of VoiceXML for voice applications contributed to reduce the proliferation of incompatible dialogue interpreters, but introduced new complexity. As a response to these issues, we designed an architecture for dialogue representation and interpretation, AdaRTE, which allows developers to layout dialogue interactions through a high level formalism that offers both declarative and procedural features. AdaRTE aim is to provide a ground for deploying complex and adaptable dialogues whilst allows the experimentation and incremental adoption of innovative speech technologies. It provides the dynamic behavior of Augmented Transition Networks and enables the generation of different backends formats such as VoiceXML. It is especially targeted to the health care context, where a framework for easy dialogue deployment could reduce the barrier for a more widespread adoption of dialogue systems. PMID- 17911879 TI - Multi-channel physiological sensing of human emotion: insights into emotion-aware computing using affective protocols, avatars and emotion specifications. AB - This paper introduces a methodology for combining multi-channel psycho physiological recordings of affective paradigms into a framework where the scientific results of such experiments are utilized in the human computer interaction context to model the computer's response based on the emotional context of the user and the situation. An affective protocol is described the results of which are expected to be combined with anthropomorphic avatars that enhance the man-machine interaction. The technological infrastructure of the later component is provided by means of XML specifications of signal descriptions and emotion recognition, as well as avatar behavior generator descriptions. PMID- 17911880 TI - A framework for cognitive monitoring using computer game interactions. AB - Many countries are faced with a rapidly increasing economic and social challenge of caring for their elderly population. Cognitive issues are at the forefront of the list of concerns. People over the age of 75 are at risk for medically related cognitive decline and confusion, and the early detection of cognitive problems would allow for more effective clinical intervention. However, standard cognitive assessments are not diagnostically sensitive and are performed infrequently. To address these issues, we have developed a set of adaptive computer games to monitor cognitive performance in a home environment. Assessment algorithms for various aspects of cognition are embedded in the games. The monitoring of these metrics allows us to detect within subject trends over time, providing a method for the early detection of cognitive decline. In addition, the real-time information on cognitive state is used to adapt the user interface to the needs of the individual user. In this paper we describe the software architecture and methodology for monitoring cognitive performance using data from natural computer interactions in a home setting. PMID- 17911881 TI - Mobile phone computing for in-situ cognitive behavioral therapy. AB - Cognitive behavioral therapy (CBT) for psychological disorders is becoming increasingly popular on the Internet. However, when using this workstation approach, components such as training and learning relaxation skills, problem solving, exposure exercises, and sleep management guidance must be done in the domestic environment. This paper describes design concepts for providing spatially explicit CBT with mobile phones. We reviewed and analyzed a set of treatment manuals to distinguish elements of CBT that can be improved and supported using mobile phone applications. The key advantage of mobile computing support in CBT is that multimedia can be applied to record, scale, and label anxiety-provoking situations where the need arises, which helps the CBT clients formulate and convey their thoughts and feelings to relatives and friends, as well as to therapists at subsequent treatment sessions. PMID- 17911882 TI - Methods for measuring the impact of health information technologies on clinicians' patterns of work and communication. AB - Evidence regarding how health information technologies influence clinical work patterns and support efficient practices is limited. Traditional paper-based data collection methods are unable to capture clinical work complexity and communication patterns. Our objective was to design and test an electronic data collection tool for work measurement studies which would allow efficient, accurate and reliable data collection, and capture work complexity. We developed software on a personal digital assistant (PDA) which captures details of nurses' work; what task, with whom, and with what; multi-tasking; interruptions and event duration. During field-testing over seven months across four hospital wards, fifty-two nurses were observed for 250 hours. Inter-rater reliability scores were maintained at over 85%. Only 1% of tasks did not match the classification developed. Over 40% of nurses' time was spent in direct care or professional communication, with 11.8% in multi-tasking. Nurses were interrupted approximately every 49 minutes. One quarter of interruptions occurred while nurses were preparing or administering medications. This approach produces data which provides greater insights into patterns of clinician's work than has previously been possible. PMID- 17911883 TI - Enhancing user acceptance of mandated mobile health information systems: the ePOC (electronic point-of-care project) experience. AB - From a clinical perspective, the use of mobile technologies, such as Personal Digital Assistants (PDAs) within hospital environments is not new. A paradigm shift however is underway towards the acceptance and utility of these systems within mobile-based healthcare environments. Introducing new technologies and associated work practices has intrinsic risks which must be addressed. This paper contends that intervening to address user concerns as they arise throughout the system development lifecycle will lead to greater levels of user acceptance, while ultimately enhancing the deliverability of a system that provides a best fit with end user needs. It is envisaged this research will lead to the development of a formalised user acceptance framework based on an agile approach to user acceptance measurement. The results of an ongoing study of user perceptions towards a mandated electronic point-of-care information system in the Northern Illawarra Ambulatory Care Team (TACT) are presented. PMID- 17911884 TI - When usage and user satisfaction differ: the case of an electronic discharge summary. AB - We describe the results of a longitudinal study regarding system use and user satisfaction before and after introduction of an electronic discharge letter application in a pediatric intensive care unit (PICU) of a German university hospital. The new discharge letter application is part of the hospital information system (HIS). The study covered an eleven month time period and used system logs as well as questionnaires including a modified questionnaire of user interaction satisfaction QUIS. We used methods which are comparable to a previous study examining a HIS based discharge letter in three departments of an Austrian hospital. In comparison we found out that user satisfaction was lower in our case. Interestingly, we noticed that in our case this was mirrored by an increasing use of the new discharge letter although there was no pressure to switch to the new HIS based discharge letter application. PMID- 17911886 TI - e-Health in Scotland: setting a baseline for stakeholder alignment. AB - Gaining knowledge of nurses' attitudes towards and understanding of various aspects of the Scottish e-Health programme is vital for needed 'stakeholder alignment'. This paper is focused on the Scottish results from a large on-line survey carried out in 2006, across the UK. Key findings, identified through analysis of both qualitative and quantitative data, are discussed. Results suggest that overall there is willingness and enthusiasm to engage and to see the modernisation of the NHS in Scotland underpinned by advanced and effective IT systems. It also shows that nurses have clear ideas about how information technology could help them and their patients in delivering safe care that enhances the visibility of the nursing contribution to care outcomes. It is argued that results help in setting a base-line from which to judge the success or otherwise of the needed e-Health clinical change management programme within clinical settings. PMID- 17911885 TI - Mapping clinicians' perceptions about computerized protocol use to an IT implementation framework. AB - Previous studies have described the determinants of successful information technology (IT) implementation. In 2003, Kukafka et al. integrated several theoretical perspectives and proposed a framework for IT implementation. This framework is applicable to IT implementation in general but lacks the identification of factors affecting adoption, which are specific to the technology under consideration. We developed and validated a model that specifically identifies factors associated with clinicians' adoption of computerized protocols. The purpose of this paper is to identify the relations between the specific factors associated with intention to use computerized protocols and the high level variables that constitute the framework proposed by Kukafka et al. Incorporation of a specific model into a general schema for IT implementation allows implementers to assess the specific individual, organizational and environmental changes required to bring about successful implementation of computerized protocols. An understanding of clinicians' perceptions specific to the technology in use will allow its seamless integration into an organization's healthcare IT plan. Strategic planning requires enhancing the framework with additional detail related to the specific technology under consideration. PMID- 17911887 TI - Usability of institutional cancer web sites: an Italian case study. AB - In order to evaluate if and to what extent Italian speaking cancer patients can benefit from information available on cancer web sites, an "in vitro" usability (ISO definition) study has been carried out. It investigated the usability of the web sites of the most representative Italian Institutions in the oncological field for the adult patients needing to find information about head and neck cancer. Specific evaluation criteria from the literature were used. The results point out some problems about accessibility, in line with other studies, and about the usefulness of the contents, in particular in the web sites of care delivery institutions: a grey present situation, but there are already grounds for significant improvement. Institutions and organizations must not waste the opportunity of being valuable sources in order to build the so called "informed patient," and the usability of their web sites could make the difference. PMID- 17911888 TI - Investigating internet use by mental health service users: interview study. AB - The internet is an increasingly important source of mental health-related information, and has the potential to be harnessed as a tool to support self-care and informed decision-making. Yet little is known about the motivations and attitudes of users. We therefore undertook a qualitative interview study with a purposive sample of mental health service users with internet experience, to explore issues with respect to mental health-related internet use. One of the prime motivations for online mental health seekers was to find experiential information from other people with similar problems. This information allowed users to know they were not alone, and to instill hope that others in the same situation had recovered. Benefits of the internet as an information source included convenience, privacy and anonymity. Problems related more to misuse of the internet rather than concerns over inaccuracy. Such qualitative work is important in an emerging research area to understand internet use better. PMID- 17911889 TI - Text characteristics of clinical reports and their implications for the readability of personal health records. AB - Through personal health record applications (PHR), consumers are gaining access to their electronic health records (EHR). A new challenge is to make the content of these records comprehensible to consumers. To address this challenge, we analyzed the text unit length, syntactic and semantic characteristics of three sets of health texts: clinical reports from EHR, known difficult materials and easy-to-read materials. Our findings suggest that EHR texts are more different from easy texts and more similar to difficult texts in terms of syntactic and semantic characteristics, and EHR texts are more similar to easy texts and different from difficult texts in regard to text unit length features. Since commonly used readability formulas focus more on text unit length characteristics, this study points to the need to tackle syntactic and semantic issues in the effort to measure and improve PHR readability. PMID- 17911890 TI - Generic screen representations for future proof systems--is it possible? Two model approach to a generic GUI. AB - Semantic interoperability should not only cover system interpretation of incoming information, but should be extended to include screen representation. This article describes a two-model approach to generate a screen representation for archetype-based information, which is inspired by the two-model approach used by openEHR for their archetypes. It provides a separation between software-related display knowledge and domain-related display knowledge and is designed with reuse of components in mind. This approach leads to a flexible GUI that can adapt not only to information structures that are not predefined within the receiving system and display them in a meaningful way, but also to novel ways of displaying the information. We are working on a proof of concept implementation to validate the approach. PMID- 17911891 TI - Speech recognition in dental software systems: features and functionality. AB - Speech recognition allows clinicians a hands-free option for interacting with computers, which is important for dentists who have difficulty using a keyboard and a mouse when working with patients. While roughly 13% of all general dentists with computers at chairside use speech recognition for data entry, 16% have tried and discontinued using this technology. In this study, researches explored the speech recognition features and functionality of four dental software applications. For each system, the documentation as well as the working program was evaluated to determine speech recognition capabilities. A comparison checklist was created to highlight each program's speech functionality. Next, after the development of charting scripts, feasibility user tests were conducted to determine if performance comparisons could be made across systems. While four systems were evaluated in the feature comparison, only two of the systems were reviewed during the feasibility user tests. Results show that current speech functionality, instead of being intuitive, is directly comparable to using a mouse. Further, systems require memorizing an enormous amount of specific terminology opposed to using natural language. User testing is a feasible way to measure the performance of speech recognition across systems and will be conducted in the near future. Overall, limited speech functionality reduces the ability of clinicians to interact directly with the computer during clinical care. This can hinder the benefits of electronic patient records and clinical decision support systems. PMID- 17911892 TI - Assessing the impact of recording quality target data on the GP consultation using multi-channel video. AB - BACKGROUND: In the UK routinely collected computerized clinical data is used to assess progress towards financially incentivised quality targets for chronic disease management including hypertension. OBJECTIVE: To develop a method for assessing the impact of recording quality target data in the clinical consultation. METHODS: Raters were trained how to rate a multi-channel video of a simulated clinical consultation for interaction between actors, computer use, non verbal communication. RESULTS: 25% of consultation time is computer use and a median of 4 to 5 items were coded per consultation mainly items related to the hypertension quality target. Intraclass correlation coefficient showed good inter rater reliability (>0.9; p<0.001). CONCLUSION: We have successfully piloted a novel technique for observing the influence of the computer on the consultation. Despite increasing computer use to record quality target data the overwhelming proportion of the consultation remains doctor patient interaction. PMID- 17911893 TI - Sustainable health systems: addressing three key areas. AB - In the modern context sustainable health systems are being developed using the newest technological and communication technologies. This is proving to be a great success for the growth of Health Informatics and healthcare improvement. However this revolution is not being reached by a lot of the world population. This paper will address the importance of closing the Digital Divide, Empowerment of health consumers and the importance of converging communications. Key areas in the development of a truly sustainable health system. PMID- 17911894 TI - Locating nursing classification schemes within health information strategies for New Zealand. AB - The potential to use classification schemes to describe and measure nursing in a country that has previously not used them as a part of practice is fraught with issues. Such is the case for New Zealand. Without nursing specific classification scheme use in the information systems of day to day function, nursing cannot validate what it does and the difference it makes to health outcomes for New Zealanders. The local use of valid and reliable classification schemes as tools to capture locally generated data that is able to be used as quality data needs to be considered alongside the national use of reliable clinical reference tools that are consistent with international standards. This may make the difference to the potential for significant contribution of nursing practice specific data to health information collections in preference to a 'one fits all' approach to user interface nursing classification scheme adoption at a local level. Tensions between a top-down approach and a locally based bottoms-up practice based approach and associated issues provide the core to this paper. PMID- 17911895 TI - SHARE, from vision to road map: technical steps. AB - We present the 'HealthGrid' initiative and briefly review work carried out in various European healthgrid projects. We report on joint work with numerous European collaborators. Since the European Commission's Information Society Technologies programme funded the first gridbased health and medical projects, the HealthGrid movement has flourished in Europe. Many projects have now been completed and 'HealthGrid' consulted a number of experts to compile and publish a 'White Paper' which establishes the foundations, potential scope and prospects of an approach to health informatics based on a grid infrastructure. With a second generation of projects now funded, the EC has commissioned the SHARE Project, a study to define a research roadmap for a 'healthgrid for Europe' as the preferred infrastructure for medical and health care projects in the European Research Area. The project explores the ways in which the healthgrid approach supports modern trends both in research in biomedicine and in healthcare, such as evidence based practice and information integration. PMID- 17911896 TI - Penetration and adoption of health information technology (IT) in Thailand's community health centers (CHCs): a national survey. AB - A universal healthcare coverage program has been implemented in Thailand since 2001 and the Thailand Ministry of Public Health (MOPH) is restructuring its health information systems to support the management of this reform. The MOPH believes that health information technology (IT) is fundamental to the development of an effective health information system, and that users' adoption of health IT is one of the most important factors to the success of health IT implementation projects. However, there is no national data available regarding the penetration and adoption of health IT in Thai community health centers (CHCs). This cross sectional survey was designed to study the penetration and adoption of health IT in the country's community health centers. A random sample of 1,607 regionally stratified CHC's from a total of 9,806 CHCs was selected. With an 82% response rate, the data showed that people who worked in CHCs were currently heavy users of health IT. They exhibited high IT acceptance and positive attitudes toward using health IT. CHCs' staff was less resistant to adopt health IT than previously anticipated. These results are similar in all of the country's geographic regions. Health IT is pervasive in CHCs across the country and penetrates all regions. PMID- 17911897 TI - The Health Informatics Center of Acadiana--informing health policymaking in post Katrina/Rita Louisiana. AB - A "healthy communities" initiative in Louisiana led to creation of the Health Informatics Center of Acadiana(HICA) at The University of Louisiana at Lafayette, in the south central United States. Since hurricanes Katrina and Rita devastated the Louisiana coast in 2005, HICA's role has taken on heightened significance. HICA identifies vulnerable populations, documents their risk factors, and evaluates interventions intended to improve community health. HICA collaborates with the Louisiana Department of Health and Hospitals and the Lafayette Community Health Consortium (LCHC), the latter formed for coordination among local healthcare providers and agencies. Both HICA and LCHC were created when "Bonne Sante a Lafayette!"--a locally developed community health improvement plan--was implemented. This paper reports on methods and experiences of HICA and LCHC, offering these as models for addressing community concerns elsewhere. Of special interest is the discussion of Louisiana HABITS, a consumer survey methodology that HICA has developed to measure healthcare access barriers, to provide information that healthcare organizations and governments need to implement workable business strategy and public policy. PMID- 17911898 TI - Informatics solutions for emergency planning and response. AB - Early informatics contributions to the emergency planning and response agenda have focused largely on surveillance of threat detection. A broader assessment of possible informatics contributions unveils that informatics can also contribute to increasing the efficiency in disaster response as well as providing a tele presence for remote medical caregivers. This presentation will explore current and future roles of informatics in emergency preparedness and response. Special challenges for data management occur with every emergency or disaster. Tracking of victims, electronic health records, and supply inventory are a few of the contributions that informatics can play during disasters. Modeling of response resources can provide the parameters for more effective decision making. Public relations reporting can be made more accurate if given the information in a timely fashion. Databases provide the infrastructure for reporting of data that can be used to manage volunteers or later be mined to determine the effectiveness of planning and response efforts. As informaticists, we have a moral obligation to contribute to the emergency response agenda worldwide. PMID- 17911899 TI - A multidiscipline conceptual framework for consumer health informatics. AB - This paper presents an idealized conceptual framework for consumer health informatics research drawing from complementary disciplines: information science and health campaign research. This synthesis is designed to provide researchers with a flexible model to evaluate current research and inform future studies. Following a description of the major components, we describe a recent evaluation of consumer perceptions of a health information system, Genetics Health Reference. This study illustrates how the framework may be applied to provide some direction and insights into ongoing consumer health informatics research. While this model represents a work in progress, we present it in support of efforts to understand the multidimensional impacts of the public's access to health information. We also discuss challenges that remain to develop a better conceptual understanding of how consumers converge on health informatics services. PMID- 17911900 TI - A Japanese model of disease management. AB - We started a disease management model, Carna, that includes two programs: one for primary prevention of lifestyle diseases and one for secondary/tertiary prevention of diabetes mellitus. These programs support the family doctor system and education for participants to allow the concept of disease management to take root in Japan. We developed a critical pathway system that can optimize health care of individual participants by matching individual status. This is the core technology of the project. Under the primary prevention program, we can perform the health check-up/ instruction tasks in the 'Tokutei Kenshin', which will start for all Japanese citizens aged 40-74 years in April 2008. In the diabetic program, Carna matches doctors and new patients, prevents patient dropout, supports detection of early-stage complications by distributing questionnaires periodically, and facilitates medical specialists' cooperation with family doctors. Carna promotes periodic medical examinations and quickly provides the result of blood tests to patients. We are conducting a study to assess the medical outcomes and business model. The study will continue until the end of 2007. PMID- 17911901 TI - Towards sustainability of health information systems: how can we define, measure and achieve it? AB - Health information systems (HIS) in their current form are rarely sustainable. In order to sustain our health information systems and with it our health systems, we need to focus on defining and maintaining sustainable Health Information System building blocks or components. These components need to be easily updatable when clinical knowledge (or anything else) changes, easily adaptable when business requirements or processes change, and easily exchangeable when technology advances. One major prerequisite for this is that we need to be able to define and measure sustainability, so that it can become one of the major business drivers in HIS development. Therefore, this paper analyses general definitions and indicators for sustainability, and analyses their applicability to HIS. We find that general 'Emergy analysis' is one possibility to measure sustainability for HIS. Based on this, we investigate major enablers and inhibitors to sustainability in a highlevel framework consisting of four pillars: clinical, technical, socio-technical, and political/business. PMID- 17911902 TI - Bermuda Triangle or three to tango: generation Y, e-health and knowledge management. AB - Generation Y workers are slowly gathering critical mass in the healthcare sector. The sustainability of future healthcare is highly dependent on this group of workers. This generation of workers loves technology and thrives in stimulating environments. They have great thirst for life-experience and therefore they move from one working environment to the other. The healthcare system has a hierarchical operational, information and knowledge structure, which unfortunately might not be the ideal ground to integrate with generation Y. The challenges ahead present a fantastic opportunity for electronic health implementation and knowledge management to flourish. Generation Y workers, however, have very different expectation of technology utilisation, technology design and knowledge presentation. This paper will argue that a clear understanding of this group of workers is essential for researchers in health informatics and knowledge management in order to provide socio-technical integrated solution for this group of future workers. The sustainability of a quality healthcare system will depend upon the integration of generation Y, health informatics and knowledge management strategies in a re-invented healthcare system. PMID- 17911903 TI - Health service organisations and professionals: an information systems model for transforming the nexus between accreditation and practice. AB - This paper presents a qualitative research approach used to generate data and theoretical insights for information systems design in a highly regulated health service organisation. Ethnographical and sociological analytic techniques were used for sensemaking in the domain, and then identifying key structures affecting the conduct of the health service. A key aspect was taking a futures perspective of possibilities for evolution of these structures. From this, a continuum model representing the nexus between accreditation and practice was developed, varying the degree of integration of information systems for organisation and individual level accreditation. The paper uses the model to discuss the implications of possibilities for design in organisations where accreditation requirements have implications for work practice and information systems design. PMID- 17911904 TI - Health informatics: an intercultural perspective. AB - Health informatics is a significant contribution to health care. It provides health professionals with powerful technologies to enhance their performance in caring for patients. The introduction of health informatics has added a new dimension in the health discourse. However, there are also issues and problems which are associated with health informatics, particularly in relation to privacy, confidentiality and data security, which are deeply embedded in culture. As privacy and confidentiality are linguistically and culturally constructed, health workers, patients and the public may have different views and exhibit different behaviours towards health informatics. The discussion of these issues is situated in an intercultural discourse. PMID- 17911905 TI - Successful systems sustaining change. AB - Much has been published on the success and particularly the failure of IT projects; still failures are commonplace. This prospective study focused from the outset on assessing risk of failure and addressing critical success factors. The aim was to apply existing methods in a challenging acute care hospital where success demanded rapid achievement of sustainable improvements in clinical and administrative processes. The implementations were part of the English National Programme for IT. The desired outcomes required the integration of accepted tools and techniques to provide a pragmatic approach to systems implementation: Lean, Six Sigma, PRINCE2 and Benefits Management. The outcome and further insights into success and failure of IT projects in healthcare are described. In particular lessons are identified related to the business need for the project and the successful achievement of the required benefits and business change. PMID- 17911906 TI - MUST--a participatory method for designing sustainable health IT. AB - Several important issues in designing sustainable health IT, such as coherent visions for change and genuine user involvement, are too often neglected or not paid enough attention to in practice. The MUST method addresses the early stages in the design of sustainable IT applications. The method highlights how those issues can be dealt with as it provides practical recommendations in terms of principles, tools and techniques. The method has proven helpful in assisting project participants to focus on and combine issues that are also crucial when designing health IT. MUST has been developed and tested in commercial settings. Here however, we illustrate the method's potential for health IT as it was recently used for the evaluation of a faulty health IT project intended to support shared care in relation to pregnancy. PMID- 17911907 TI - Change management and the sustainability of health ICT projects. AB - The development of the electronic health record (EHR) is a strategic and important enabler of the delivery of integrated healthcare. As each innovative aspect of the EHR is implemented in New Zealand, long-term success is essential for its overall sustainability on the national scale. How we achieve this success is dependent upon how people adapt to the changes brought about by the implementation of these innovations. The transition period of the change process we follow during this adaptation is characterized by a capability crisis, in which we tend to predict failure in our attempts to make the changes to which we are committed. This could be a signal of the first step toward sustainable change as people adapt to changed processes, technology and relationships. Once we have mastered the incremental changes brought about by health ICT projects for the implementation of the EHR, we are able to connect health services by means of the same EHR and provide enabled, sustainable integrated healthcare. PMID- 17911908 TI - A sustainability view on the EPR system of N.N. Burdenko Neurosurgical Institute. AB - This paper aims at the analysis of the "sustainability status" of an electronic patient record system developed at the Medical Informatics Laboratory of the N. N. Burdenko Neurosurgical Institute (EPR/NSI). It includes some of the principles that allowed a small team of developers to create a sustainable EPR system for a large medical institution with complicated diagnostic and treatment processes. PMID- 17911909 TI - Predicting coronary artery disease with medical profile and gene polymorphisms data. AB - Coronary artery disease (CAD) is a main cause of death in the world. Finding cost effective methods to predict CAD is a major challenge in public health. In this paper, we investigate the combined effects of genetic polymorphisms and non genetic factors on predicting the risk of CAD by applying well known classification methods, such as Bayesian networks, naive Bayes, support vector machine, k-nearest neighbor, neural networks and decision trees. Our experiments show that all these classifiers are comparable in terms of accuracy, while Bayesian networks have the additional advantage of being able to provide insights into the relationships among the variables. We observe that the learned Bayesian Networks identify many important dependency relationships among genetic variables, which can be verified with domain knowledge. Conforming to current domain understanding, our results indicate that related diseases (e.g., diabetes and hypertension), age and smoking status are the most important factors for CAD prediction, while the genetic polymorphisms entail more complicated influences. PMID- 17911910 TI - Towards a top-domain ontology for linking biomedical ontologies. AB - In this paper we present the ongoing development and extension work on BioTop--a top-domain ontology for linking biomedical domain ontologies. We start by making the case for the application of a common ontology to interface independent biomedical domain ontologies by introducing a set of more general classes. Then we briefly depict the relation of BioTop to the GENIA ontology as starting point of its initial developement. Afterwards we propose our distinction of ontologies into top, top-domain and domain ones and describe our approach to the integration of the top ontology BFO into BioTop. Then we present our plans to join the OBO and OBO Foundry repository of ontologies and list its admission principles in relation to our ontology. Some actual BioTop interface classes are shown subsequently. We conclude by detailing on some planned BioTop usages in the area of BioNLP and cancer research and show some further intended improvements. PMID- 17911911 TI - The molecular medicine informatics model (MMIM). AB - In 2005 a major collaboration in Melbourne, Australia successfully implemented a major medical informatics infrastructure. The convergence of life sciences, healthcare, and information technology is now driving research into the fundamentals of disease causation and toward tailoring individualized treatment. The Molecular Medicine Informatics Model (MMIM) is a 'virtual' research repository of clinical, laboratory and genetic data sets. Integrated data, physically located within independent hospital and research organisations can be searched and queried seamlessly via a federated data integrator. Researchers must gain authorisation to access data, and obtain permission from the data owners before the data can be accessed. The legal and ethical issues surrounding the use of this health data have been addressed so data complies with privacy requirements. The MMIM platform also record links individual cases across multiple institutions and multiple clinical specialties. Significant research outcomes in epilepsy and colorectal cancer have already been enabled by the MMIM research platform. The infrastructure of MMIM enables discovery research to be accessible via the Web with security, intellectual property and privacy addressed. PMID- 17911912 TI - Cancer genomics object model: an object model for multiple functional genomics data for cancer research. AB - The development of functional genomics including transcriptomics, proteomics and metabolomics allow us to monitor a large number of key cellular pathways simultaneously. Several technology-specific data models have been introduced for the representation of functional genomics experimental data, including the MicroArray Gene Expression-Object Model (MAGE-OM), the Proteomics Experiment Data Repository (PEDRo), and the Tissue MicroArray-Object Model (TMA-OM). Despite the increasing number of cancer studies using multiple functional genomics technologies, there is still no integrated data model for multiple functional genomics experimental and clinical data. We propose an object-oriented data model for cancer genomics research, Cancer Genomics Object Model (CaGe-OM). We reference four data models: Functional Genomic-Object Model, MAGE-OM, TMAOM and PEDRo. The clinical and histopathological information models are created by analyzing cancer management workflow and referencing the College of American Pathology Cancer Protocols and National Cancer Institute Common Data Elements. The CaGe-OM provides a comprehensive data model for integrated storage and analysis of clinical and multiple functional genomics data. PMID- 17911913 TI - Automatic pedigree reconstruction for genetic studies in isolated populations. AB - This paper describes a tool implemented to automatically reconstruct the pedigree of an isolated population of Northern Italy with the aim of supporting genetic studies. The goal of such studies is to analyze genealogic, clinical and genetic data for genetic dissection of complex diseases. In this context the reconstruction of the population pedigree is fundamental to verify that such population is a genetic isolate and obtain the parental relationships among the individuals participating to the study. The algorithm presented in the paper, from heterogeneous data sources (demographic municipal and parish archives and other data sources), derives the pedigree applying several heuristic rules in a predefined order. One of the main difficulties in performing such task stands in the "record linkage" process that requires the definition of a sufficiently general strategy for managing the ambiguities caused by missing or imprecise/erroneous input data. The paper, finally, presents and discusses the preliminary results obtained by reconstructing the pedigree of four villages from the data collected during the first eighteen months of project. PMID- 17911914 TI - Enhancing the quality of phylogenetic analysis using fuzzy hidden Markov model alignments. AB - Any effective phylogeny inference based on molecular data begins by performing efficient multiple sequence alignments. So far, the Hidden Markov Model (HMM) method for multiple sequence alignment has been proved competitive to the classical deterministic algorithms with respect to phylogenetic analysis; nevertheless, its stochastic nature does not help it cope with the existing dependence among the sequence elements. This paper deals with phylogenetic analysis of protein and gene data using multiple sequence alignments produced by fuzzy profile Hidden Markov Models. Fuzzy profile HMMs are a novel type of profile HMMs based on fuzzy sets and fuzzy integrals, which generalize the classical stochastic HMM by relaxing its independence assumptions. In this paper, alignments produced by the fuzzy HMM model are used in phylogenetic analysis of protein data, enhancing the quality of phylogenetic trees. The new methodology is implemented in HPV virus phylogenetic inference. The results of the analysis are compared against those obtained by the classical profile HMM model and depict the superiority of the fuzzy profile HMM in this field. PMID- 17911915 TI - Determining transcription factor activity from microarray data using Bayesian Markov chain Monte Carlo sampling. AB - Many biological processes rely on remodeling of the transcriptional response of cells through activation of transcription factors. Although determination of the activity level of transcription factors from microarray data can provide insight into developmental and disease processes, it requires careful analysis because of the multiple regulation of genes. We present a novel approach that handles both the assignment of genes to multiple patterns, as required by multiple regulation, and the linking of genes in prior probability distributions according to their known transcriptional regulators. We demonstrate the power of this approach in simulations and by application to yeast cell cycle and deletion mutant data. The results of simulations in the presence of increasing noise showed improved recovery of patterns in terms of chi2 fit. Analysis of the yeast data led to improved inference of biologically meaningful groups in comparison to other techniques, as demonstrated with ROC analysis. The new algorithm provides an approach for estimating the levels of transcription factor activity from microarray data, and therefore provides insights into biological response. PMID- 17911916 TI - Ensemble stump classifiers and gene expression signatures in lung cancer. AB - Microarray data sets for cancer tumor tissue generally have very few samples, each sample having thousands of probes (i.e., continuous variables). The sparsity of samples makes it difficult for machine learning techniques to discover probes relevant to the classification of tumor tissue. By combining data from different platforms (i.e., data sources), data sparsity is reduced, but this typically requires normalizing data from the different platforms, which can be non-trivial. This paper proposes a variant on the idea of ensemble learners to circumvent the need for normalization. To facilitate comprehension we build ensembles of very simple classifiers known as decision stumps--decision trees of one test each. The Ensemble Stump Classifier (ESC) identifies an mRNA signature having three probes and high accuracy for distinguishing between adenocarcinoma and squamous cell carcinoma of the lung across four data sets. In terms of accuracy, ESC outperforms a decision tree classifier on all four data sets, outperforms ensemble decision trees on three data sets, and simple stump classifiers on two data sets. PMID- 17911917 TI - From "glycosyltransferase" to "congenital muscular dystrophy": integrating knowledge from NCBI Entrez Gene and the Gene Ontology. AB - Entrez Gene (EG), Online Mendelian Inheritance in Man (OMIM) and the Gene Ontology (GO) are three complementary knowledge resources that can be used to correlate genomic data with disease information. However, bridging between genotype and phenotype through these resources currently requires manual effort or the development of customized software. In this paper, we argue that integrating EG and GO provides a robust and flexible solution to this problem. We demonstrate how the Resource Description Framework (RDF) developed for the Semantic Web can be used to represent and integrate these resources and enable seamless access to them as a unified resource. We illustrate the effectiveness of our approach by answering a real-world biomedical query linking a specific molecular function, glycosyltransferase, to the disorder congenital muscular dystrophy. PMID- 17911918 TI - Building a research model for human genetic variation knowledge management. AB - Organizational knowledge management (KM) research studies the nature of knowledge, the scope of KM, the factors and mechanisms that affect KM outcomes, as well as theoretical KM frameworks. This paper discusses the implications of past studies for the KM efforts in the human genetic variation (HGV) research domain and presents a HGV-KM research model. This model identifies the context of HGV KM studies, the predispositions and factors that may impact KM outcomes, and important KM processes. It also represents the relationships among these issues. Applying the model, further studies will point the way for improved capture and dissemination of HGV knowledge from routine HGV research activities to contribute to the global genetics knowledge base. PMID- 17911919 TI - ECTracker--an efficient algorithm for haplotype analysis and classification. AB - This work aims at discovering the genetic variations of hemophilia A patients through examining the combination of molecular haplotypes present in hemophilia A and normal local populations using data mining methods. Data mining methods that are capable of extracting understandable and expressive patterns and also capable of making predictions based on inferences made on the patterns were explored in this work. An algorithm known as ECTracker is proposed and its performance compared with some common data mining methods such as artificial neural network, support vector machine, naive Bayesian, and decision tree (C4.5). Experimental studies and analyses show that ECTracker has comparatively good predictive accuracies in classification when compared to methods that can only perform classification. At the same time, ECTracker is also capable of producing easily comprehensible and expressive patterns for analytical purposes by experts. PMID- 17911920 TI - A dynamic query system for supporting phenotype mining in genetic studies. AB - This paper describes an information technology infrastructure aimed at supporting translational bioinformatics studies that require joint management of phenotypic and genotypic data. In particular, we integrated an electronic medical record with an open-source environment for data mining to create a flexible and easy to use query system aimed at supporting the discovery of the most frequent complex traits. We propose a logical formalization to define the phenotypes of interest; this is translated into a graphical interface that allows the user to combine different conditions relative to the electronic medical record data (e.g., the presence of a particular pathology). The phenotypes are then stored in a multidimensional database. Then, the data mining system engine reads the filtered data from the database and executes dynamic queries for analyzing phenotypic data, presenting the results in a multidimensional format through a simple web interface. The system has been applied in a study on genetically isolated individuals, the Val Borbera project. PMID- 17911921 TI - Identifying QT prolongation from ECG impressions using natural language processing and negation detection. AB - Electrocardiogram (ECG) impressions provide significant information for decision support and clinical research. We investigated the presence of QT prolongation, an important risk factor for sudden cardiac death, compared to the automated calculation of corrected QT (QTc) by ECG machines. We integrated a negation tagging algorithm into the KnowledgeMap concept identifier (KMCI), then applied it to impressions from 44,080 ECGs to identify Unified Medical Language System concepts. We compared the instances of QT prolongation identified by KMCI to the calculated QTc. The algorithm for negation detection had a recall of 0.973 and precision of 0.982 over 10,490 concepts. A concept query for QT prolongation matched 2,364 ECGs with precision of 1.00. The positive predictive value of the common QTc cutoffs was 6-21%. ECGs not identified by KMCI as prolonged but with QTc>450ms revealed potential causes of miscalculated QTc intervals in 96% of the cases; no definite concept query false negatives were detected. We conclude that a natural language processing system can effectively identify QT prolongation and other cardiac diagnoses from ECG impressions for potential decision support and clinical research. PMID- 17911922 TI - A comparison of supervised classification methods for auditory brainstem response determination. AB - The ABR is commonly used in the Audiology clinic to determine and quantify hearing loss. Its interpretation is subjective, dependent upon the expertise and experience of the clinical scientist. In this study we investigated the role of machine learning for pattern classification in this domain. We extracted features from the ABRs of 85 test subjects (550 waveforms) and compared four complimentary supervised classification methods: Naive Bayes, Support Vector Machine Multi Layer Perceptron and KStar. The Abr dataset comprised both high level and near threshold recordings, labeled as 'response' or 'no response' by the human expert. Features were extracted from single averaged recordings to make the classification process straightforward. A best classification accuracy of 83.4% was obtained using Naive Bayes and five relevant features extracted from time and wavelet domains. Naive Bayes also achieved the highest specificity (86.3%). The highest sensitivity (93.1%) was obtained with Support Vector Machine-based classification models. In terms of the overall classification accuracy, four classifiers have shown the consistent, relatively high performance, indicating the relevance of selected features and the feasibility of using machine learning and statistical classification models in the analysis of ABR. PMID- 17911923 TI - Non-linear analysis for the sleepy drivers problem. AB - The problem addressed in this work is sleepiness during driving, which often leads to accidents in the streets. Experiments with sleepy drivers took place and the EEG data were analysed in terms of non-linear methods. Sample entropy and phase synchronization variations were investigated within the signal sections corresponding to "driving events", i.e. driving mistakes or loss of control, as well as to periods of drowsiness and sleepiness, as compared to the periods of normal driving. Decreased sample entropy, indicating loss of complexity, and an increased phase synchronisation have been found in the preliminary study presented. The results are encouraging towards developing an alerting system for predicting and preventing driving accidents. PMID- 17911924 TI - Identification and genotype related classification of children with long QT syndrome using 24 h Holter recordings. AB - The long QT syndrome (LQTS) belongs to the family of hereditary diseases and can cause life-threatening arrhythmias and leads to sudden cardiac death. Mutations on six genes are responsible for changes in the electrophysiological properties of myocardial cells that are involved in the repolarization phase. In the surface ecg this is expressed by a prolonged QT interval and genotypespecific shapes for the T-Wave. The aim of the study was to find parameters that quantify properties of the repolarization phase which can be used in addition to the established Schwartz score in the process of diagnosing LQTS. Furthermore, ecg features were evaluated for the separation of the LQT subtypes LQT1, LQT2 and LQT3. The combination of the features PtA50 and QTc yielded with 93% sensitivity and 100% specificity the best results in the field of patient identification. Despite the small dataset consisting of 14 patients that was available for the second aim, the achieved results for the morphology indices motivate further research in this field. PMID- 17911925 TI - Temporal abstraction and data mining with visualization of laboratory data. AB - To analyze the laboratory data by data mining, user-centered universal tools have not been available in medicine. We analyzed 1,565,877 laboratory data of 771 patients with viral hepatitis in order to find the difference of the temporal changes in laboratory test data between Hepatitis B and Hepatitis C by the combination of temporal abstraction and data mining. The data for one patient is temporal for more than 5 years. After pretreatment the data was converted to abstract patterns and then selected into sets of data combination and rules to identify Hepatitis B or C by D2MS and LUPC which were originally produced by ourselves. Not only data pattern, but also temporal relations were considered as a part of the rules. In the course of evaluating the results by domain experts, even though there were not so remarkable hypotheses, visualization tools made it easier for them to understand the relations of the complicated rules. PMID- 17911926 TI - Automated interpretation of optic nerve images: a data mining framework for glaucoma diagnostic support. AB - Confocal Scanning Laser Tomography (CSLT) techniques capture high-quality images of the optic disc (the retinal region where the optic nerve exits the eye) that are used in the diagnosis and monitoring of glaucoma. We present a hybrid framework, combining image processing and data mining methods, to support the interpretation of CSLT optic nerve images. Our framework features (a) Zernike moment methods to derive shape information from optic disc images; (b) classification of optic disc images, based on shape information, to distinguish between healthy and glaucomatous optic discs. We apply Multi Layer Perceptrons, Support Vector Machines and Bayesian Networks for feature sub-set selection and image classification; and (c) clustering of optic disc images, based on shape information, using Self-Organizing Maps to visualize sub-types of glaucomatous optic disc damage. Our framework offers an automated and objective analysis of optic nerve images that can potentially support both diagnosis and monitoring of glaucoma. PMID- 17911927 TI - Intelligent querying and exploration of multiple time-oriented medical records. AB - Querying and analyzing multiple time-oriented patient data is a key task during medical research, clinical trials or the assessment of the quality of therapy. In this paper, we present several aspects of the VISITORS system, which includes knowledge-based tools for graphical querying and exploration of multiple longitudinal patient records. We focus on the syntax and semantics of the knowledge-based aggregation query language for multiple time-oriented patient records, and on the graphical query-construction interface. The query language assumes an underlying computational method for deriving meaningful abstractions from single and multiple patient records, such as we had previously developed. The aggregation query language enables population querying using an expressive set of constraints. By using our underlying temporal mediator architecture, the time needed to answer typical temporal-abstraction aggregation queries on databases of 1000 to 10,000 patients was reasonable. PMID- 17911928 TI - Analyzing web log files of the health on the net HONmedia search engine to define typical image search tasks for image retrieval evaluation. AB - Medical institutions produce ever-increasing amount of diverse information. The digital form makes these data available for the use on more than a single patient. Images are no exception to this. However, less is known about how medical professionals search for visual medical information and how they want to use it outside of the context of a single patient. This article analyzes ten months of usage log files of the Health on the Net (HON) medical media search engine. Key words were extracted from all queries and the most frequent terms and subjects were identified. The dataset required much pre-treatment. Problems included national character sets, spelling errors and the use of terms in several languages. The results show that media search, particularly for images, was frequently used. The most common queries were for general concepts (e.g., heart, lung). To define realistic information needs for the ImageCLEFmed challenge evaluation (Cross Language Evaluation Forum medical image retrieval), we used frequent queries that were still specific enough to at least cover two of the three axes on modality, anatomic region, and pathology. Several research groups evaluated their image retrieval algorithms based on these defined topics. PMID- 17911929 TI - Improving computer aided disease detection using knowledge of disease appearance. AB - The accurate quantification of disease patterns in medical images allows radiologists to track the progress of a disease. Various computer vision techniques are able to automatically detect different patterns that appear on images. However, classical pattern detection approaches do not perform satisfactorily on medical images. The problem is that texture descriptors, alone, do not capture information that is pertinent to medical images, i.e. the disease appearance and distribution. We present a method that uses knowledge of anatomy and specialised knowledge about disease appearance to improve computer-aided detection. The system has been tested on detecting honeycombing - a diffuse lung disease pattern in HRCT images of the lung. The results show that the proposed knowledge guided approach improves the accuracy of honeycombing detection. A paired t-test, shows the improvement in accuracy to be statistically significant (p<0.0001). PMID- 17911930 TI - MR atlas for articular cartilage morphology: potential to detect shape differences. AB - An atlas of the cartilage was created using free form transformation of MR images of the cartilage from 20 subjects. The deformation required to move each voxel to its corresponding location in the atlas is used to determine the differences in shape between cartilages of subjects in a population. Based on these active shape models, it is possible to localize regions of high morphological variance in population cohorts. The atlas, reported here, is based on 20 male subjects; ten symptomatic of arthritis and ten asymptomatic. The active shape models based on this atlas show regions of high morphological variance corresponding to cartilage thinning in the arthritic group. This method has the potential to differentiate between normal and arthritic population groups by detecting subtle morphological changes in articular cartilage. PMID- 17911931 TI - Automatic image modality based classification and annotation to improve medical image retrieval. AB - Medical image retrieval can play an important role for diagnostic and teaching purposes in medicine. Image modality is an important visual characteristic that can be used to improve retrieval performance. Many test and online collections do not contain information about the image modality. We have created an automatic image classifier for both grey-scale and colour medical images. We evaluated the performance of the two modality classifiers, one for grey-scale images and the other for colour images on the CISMeF and the ImageCLEFmed 2006 databases. Both classifiers were created using a neural network architecture for learning. Low level colour and texture based feature vectors were extracted to train the network. Both classifiers achieved an accuracy of >95% on the test collections that they were tested on. We also evaluated the performance of these classifiers on a selection of queries from the ImageCLEFmed 2006. The precision of the results was improved by using the modality classifier to resort the results of a textual query. PMID- 17911932 TI - Quantification of myocardial perfusion for CAD diagnosis. AB - We introduce a computer based algorithm for objective quantification of myocardial perfusion to support the diagnosis of cad patients. This new method is based on conventional cine angiographic films. In order to achieve maximal quality of the digital subtraction angiography images, the sequence is synchronized with the ECG. Optionally, the digital images can be motion compensated by a two step matching method. The spatio-temporal spread of blood, or the so-called blush, through the microvasculature to the myocardium--indicated by dye injection--represents a characteristic pattern for the myocardial perfusion. This dynamic temporal pattern is characterized by typical features as the maximal value of blush intensity, of increase and of decrease velocity which correspond with the different phases of flooding in and washout. On the basis of 100 different temporal blush profiles, an algorithm is established which classifies the acquired blush patterns into 4 different grades. PMID- 17911933 TI - Enabling the safe and effective implementation of health informatics systems- validating rolling out the ECDL/ICDL health supplement. AB - Sound understanding by end-users (health professionals and support staff) of key e-health principles and challenges is essential for the safe, effective, and sustainable use of health informatics systems. This is separate from, and ideally a precursor to, training on a specific system. However, hitherto this aspect has been little understood or addressed. Over the last few years, the concept of a customised Health Supplement to the well-established European/International Computer Driving Licence has progressed from idea to reality, through initial support for consultation by the then UK NHS Information Authority, followed by establishment of an international Expert Group by the global ECDL/ICDL Foundation. As a result, the ECDL/ICDL Foundation has developed a formal syllabus. This paper reports successful piloting, and progress in the development of local training and testing materials and national implementation plans, in three countries on two continents. PMID- 17911934 TI - A multi-method approach to assessing health information systems end users' training needs. AB - Many existing training programs for Health Information Systems (HIS) do not adequately satisfy end users' needs nor meet training objectives. This is because they do not envisage the problems that users may encounter when performing specific tasks. Therefore the first priority for the development of an effective training program is to precisely assess the end users' training needs, a process called Training Needs Assessment (TNA). Applying traditional approaches for TNA, such as interviews or surveys alone, however, may be insufficient because they are limited in their capacity to reveal the cognitive processes of end users. Usability testing, with its ability to gather data about human computer interaction, overcomes the deficiencies of these traditional approaches. This paper proposes a multi-method approach, which combines usability testing together with traditional methods, such as interviews or questionnaire surveys to assess HIS end users' training needs. This innovative method is able to precisely reveal the training needs for different levels of HIS users. A case study, which applied this method to assess the training needs for users of a nursing information system demonstrates its feasibility. PMID- 17911935 TI - Making health informatics competencies useful: an applied health informatics competency self-assessment system. AB - Years ago we undertook to define Health Informatics (HI) competencies. This effort resulted in the creation of a document that articulated HI roles, the challenges faced by HI professionals, the high-level tasks that they needed to undertake to address these challenges and the competencies (skills, knowledge, and experience) they needed to complete these tasks. Unfortunately, in so doing we created what is arguably the most boring book in history, shoes contents are very difficult to extract, use, maintain and improve. We report here the completion of a pilot of a system that we believe corrects this situation. It is a webbased tool that incorporates all of the material, from roles to detailed competencies, enabling them to be accessed and used for a variety of purposes, the most notable of which is professional self-assessment. PMID- 17911936 TI - E-learning for students in their first year: a French experimentation at the medical school of Grenoble. AB - A local study carried out in the Medical School of Grenoble shows that teaching in the first year in medicine studies satisfies neither the students, nor the teachers. The Faculty of Medicine of Grenoble decided to set up a reform in order to offer a high quality education. This reform leads to a complete reorganization of the curriculum and to the intensive use of new information and communication technologies of information, in particular, the use of multimedia documents. The communication and information technologies team of the Faculty of Medicine of Grenoble carried out an innovating and daring reform to start at the academic year 2006-2007. The new course is built on three activities: self learning on multi-media resources, meetings with teachers for questions-answers sessions and tutorials animated by older students. This article reports the first results for this successful project. In the academic year 2006-2007, are concerned 1290 students, 40 teachers and 8 disciplines. PMID- 17911937 TI - E-learning at Porto Faculty of Medicine. A case study for the subject 'Introduction to Medicine'. AB - The main objective of the Introduction to Medicine (IM) subject of the first year of the Medical Course at the Faculty of Medicine of the University of Porto is to provide students with a first contact with the areas of Biostatistics, Medical Informatics, Bioethics and the History of Medicine in the belief that they will be better prepared to learn, research, evaluate, share and decide within their practice. This paper presents a case study that describes how the subject IM is organized and how the b-learning tool (Moodle) is used to correct and grade the students' work. From the 239 students registered to attend the Introduction to Medicine subject 12% failed. The average grade among the successful students was 16 (out of 20). In the previous academic year only 2% of the students failed. However, among the successful students, the average grade was inferior (15 out of 20). The e-learning model that was described in this paper was successful because the results show that the students that made use of the Moodle got better grades. PMID- 17911938 TI - Designing m-learning for junior registrars--activation of a theoretical model of clinical knowledge. AB - The MINI-project aims at supporting junior registrars in the learning process of how to utilize their theoretical knowledge from Medical School in everyday clinical reasoning and practice. Due to the nature of the work--concurrent moving, learning and producing--we designed an m-learning application. This paper introduces the possibilities and challenges for design of the m-learning application based on a) analytical findings on learning and mobility as derived from the design case--an emergency medical ward b) theoretical perspectives on medical knowledge, and c) presentation of the design of an m-learning application. The design process was based on user-driven innovation and the paper discusses considerations on how to combine user-drive and generic models. PMID- 17911939 TI - Evaluation and assessment of the online postgraduate critical care nursing course. AB - During challenging times facing the health service, strategies for sustaining further education for nurses in highly specialised areas call for alternate means for learning. Nurses, who were accustomed to traditional methods of learning and had no formal computer training as part of their curriculum, are now being confronted with new methods of learning. Evaluation of the effectiveness of a newly developed postgraduate critical care course delivered online for nurses was examined. A pre test and post test of 16 participants were conducted. Participants found coursework intellectually stimulating and their preference to learn from websites demonstrated the effect size (tau-b=.677) to be significant at the 0.01 level. The pre and post test results from the knowledge assessment tool indicated an advancement of mean test scores and at a significant difference value of p=.055. Ninety four percent of the participants agreed that they were able to integrate their learning from the coursework towards their clinical practice. Improvement in nurses critical care knowledge impacts positively on delivery of safe and effective health care. PMID- 17911941 TI - Data mining results from an electronic clinical log for nurse practitioner students. AB - Traditional techniques for collecting data on clinical experiences have been greatly flawed. Data cannot be easily collected in real time to make programmatic or placement changes "on the fly". Furthermore, it is difficult to look at data across students, specialty areas, and years because the data is typically not in a digital format. In response to this problem, the Vanderbilt University School of Nursing has created a web/PDA based clinical log to document the kinds of clinical experiences the students are having. Since our initial report, three years ago, we have collected three years worth of data, over 220,000 different patient encounters. This past year the data has been very complete, giving a full picture of the types of experiences the students are having. Our faculty have begun to analyze the data in the clinical log to examine the kind of experiences the students are having and to make programmatic changes and placement adjustments in real time. In general, the results supported that students in the various specialties managed patients and performed services appropriate to their specialty. Patients varied in ages, ethnic groups, payment sources, and medical diagnoses. Students did progress from an observer role to a more independent role in either a linear fashion or in a more biphasic mode with an increase in the observer role at the start of a new semester. PMID- 17911940 TI - Development and evaluation of a PDA-based decision support system for pediatric depression screening. AB - Depression is under recognized in a variety of pediatric settings. The purpose of this paper is to describe the development and initial evaluation of a personal digital assistant (PDA)-based decision support system (DSS) for pediatric depression screening in ages 8 to 18 years of age by pediatric advanced practice nurse (APN) students. Three aspects are described: selection of depression screening instrument; integration of the instrument into the PDA; and quantitative (usage) and qualitative (focus group) evaluation. Only one third of eligible patients were screened. Twenty percent of those screened were identified as at risk for mood disorder. The barriers to screening identified through focus groups included a lack of time, knowledge, intervention protocol, referral resources, PDA usability issues, preceptor motivation and comfort, as well as perceived or real cultural barriers. Suggestions for educational, research, and interventions to integrate clinical based PDA-based screening are discussed. PMID- 17911942 TI - Phase I implementation of an academic medical record for integrating information management competencies into a nursing curriculum. AB - This paper is the report of the first phase of a case study from the University of Utah to help students and faculty integrate electronic information management into the nursing curriculum. Cerner AES, a live-production clinical information system with an academic overlay, has been implemented into the first semester of an undergraduate nursing program. A consortium of schools that use Cerner AES collaborate in the design and implementation of forms used by students. The consortium also allows members to share strategies for using the system. By using the system students are developing needed informatics competencies for beginning level nurses. The paper discusses the implementation strategies used and initial results of this project. Plans for expanding the project throughout the nursing curriculum are also presented. PMID- 17911943 TI - ROC van Twente: nursing education in care and technology. AB - The ROC van Twente offers nursing education at the diploma level (MBO), and is innovating the program to include a major/minor structure for education about care and technology. In order to achieve this, a new position was created: the Master Docent, Care and Technology. The task of the master docent includes development of education for nursing about technology, multidisciplinary cooperation, and service to health care institutions among others. The first development concerns a module about electronic patient records, standards, and semantic interoperability for continuity of care. The module is delivered to nursing students and to students from the information technology department, who work jointly in 'development teams'. This paper describes the background, the development of the educational material and program, and the core content of the module. The core content are the care information models that link clinical materials with health care information standards. The program has started end November 2006. At the Medinfo 2007 conference the results of the course for the first group of about 40 students will be presented. PMID- 17911944 TI - Multiple measures of provider participation in Internet delivered interventions. AB - Evaluation of Internet-delivered continuing education for health care providers requires appropriate consideration of their level of participation. To fully assess participation requires multidimensional measures, including factors such as the volume of participation (page views), frequency (visits), variety (components accessed by each provider), and duration (months of activity). We calculated crude and refined (adjusted for study design) measures and then compared these measures across three longitudinal Internet-delivered continuing education to health care providers (N=429). We found that participation varied across study, varied by factor and varied by specific measure. Correlation between crude and refined measures within a factor and across factors differed significantly. Participation assessment of internet-delivered interventions varies by the selection of measure and factor. Further research assessing the potential for these measures to predict intervention effectiveness is needed. PMID- 17911946 TI - Importance of public health informatics: a survey of public health schools and graduate programs in the United States. AB - This paper examines the importance of data, information, and informatics to public health practice. Forty public health academicians from 40 schools and graduate programs of public health were interviewed. All agreed that informatics was important to public health practice. A qualitative analysis of their comments revealed their beliefs on the importance of informatics skills and knowledge to the practice of public health. The resulting comment groups varied from "some skills are more important than others" to "need all the skills." Eight "importance" comment groups were formed: 1) skills for all professionals; 2) some skills more than others; 3) yes, they need all the skills; 4) skills to become better practitioners; 5) usefulness to practitioners; 6) communication with public; 7) they're [the public] are depending on us; and 8) the future. PMID- 17911945 TI - Medical students' knowledge and perceptions of e-health: results of a study in Sri Lanka. AB - The present study investigates the knowledge, perceptions and attitudes of medical students in Sri Lanka in regard to e-health. We also examined the barriers which impede them to develop knowledge and skills in e-health within their medical curriculum. A questionnaire focusing on the knowledge, attitudes and expectations of medical students towards e-health was distributed to all final year students (n=136) at the Faculty of Medical Sciences, Sri Jayewardenepura University, Sri Lanka. Response rate was 74%. 43% of respondents stated that they were familiar with the term e-health. 51% rated their knowledge of e-health applications as minimal. 88% admitted that they had no e-health education or training of any kind. Over 80% of all respondents thought that e health had an important role to play in the current and future health sector, particularly in developing countries. Our survey revealed that respondents had very poor access to computers and Internet use was rare. 77% of respondents admitted that they were not provided with systematic knowledge and skills in e health through their medical curriculum and identified the absence of formal education in e-health as a serious shortcoming. PMID- 17911947 TI - Educating medical students as competent users of health information technologies: the MSOP data. AB - As more health information technologies become part of the health care environment, the need for physicians with medical informatics competencies is growing. In 2006, a survey was created to determine the degree to which the Association of American Medical College's Medical School Objectives Project (MSOP) medical informatics competencies had been incorporated into medical school curricula in the United States. METHODS: a web-based tool was used to create the survey; medical education deans or their designees were requested to complete the survey. Analysis focused on the clinician, researcher, and manager roles of physicians. RESULTS: Seventy usable surveys were returned. Many of the objectives were stated in the schools' respective curricula and the competencies were being evaluated. However, only a few schools taught and assessed the medical informatics objectives that required interaction with health information. CONCLUSION: To insure that physicians have the knowledge, skills, and attitudes to effectively and efficiently interact with today's health information technologies, more medical informatics concepts need to be included and assessed in all undergraduate medical education curricula in the United States. PMID- 17911948 TI - Establishing a national resource: a health informatics collection to maintain the legacy of health informatics development. AB - This case study report of the establishment of a national repository of multi media materials describes the creation process, the challenges faced in putting it into operation and the opportunities for the future. The initial resource has been incorporated under standard library and knowledge management practices. A collaborative action research method was used with active experts in the domain to determine the requirements and priorities for further development. The National Health Informatics Collection (NatHIC) is now accessible and the further issues are being addressed by inclusion in future University and NHS strategic plans. Ultimately the Collection will link with other facilities that contribute to the description and maintenance of effective informatics in support of health globally. The issues raised about the National Health Informatics Collection as established in the UK have resonance with the challenges of capturing the overall historic development of an emerging discipline in any country. PMID- 17911949 TI - The EIPEN project: promoting interprofessional education in health professions. AB - The Leonardo project under the name European Interprofessional Education Network (EIPEN) in health and social care, has been dealing with the challenges of Interprofessional Education (IPE). The EIPEN project tries to develop a transnational network of universities and employers in the six participating countries and at the same time to promote good practices in Interprofessional LearningTeaching in healthsocial care. IPE provides opportunities for students and practitioners to learn with, from and about each other during qualifying and post-qualifying training and in their practice. IPE in health and social care includes the education and training of practitioners in human and animal medicine, dentistry, nursing, physiotherapy, occupational therapy, pharmacy and all other health professions including public and environmental health and health promotion, and social work. The outcomes of the EIPEN Project will provide means, material and guidelines for the enhancement of professional education in the multidisciplinary field of Health Informatics. PMID- 17911950 TI - Building ICT capabilities for clinical work in a sustainable healthcare system: approaches to bridging the higher education learning and teaching gap. AB - There is a recognised gap in information and communications technology (ICT) learning and teaching in higher education for entry-level healthcare professionals. This paper proposes a research model for understanding the dimensions of this gap. We describe methodological approaches to understanding present practices, identifying levers for change and learning by doing. We discuss issues faced in getting started and sustaining momentum on the research that is an essential prerequisite to effectively build the ICT capacity required by the clinical workforce in a sustainable healthcare system. PMID- 17911951 TI - The development of an online clinical log for advanced practice nursing students: a case study. AB - Three years ago at the Medinfo conference a prototype version of a clinical log for nursing students at Vanderbilt University was demonstrated. The purpose of the log is to document the types of clinical experiences the students are participating in as part of their academic program. We collected log data during that first year and received ongoing formative feedback from both students and faculty regarding its current feature set and desirable features for future implementations. Most of the requested new features have now been implemented. This paper describes some of the latest features of the clinical log, the advantages and disadvantages of ongoing development versus acquiring commercial products, and the procedures and results we have put in place to gather from faculty and students the features they want to see in the log. This paper also documents some of the data from early data mining. PMID- 17911952 TI - Personalized case driven parental education informatics in the NICU. AB - Neonatal Intensive Care Units (NICUs) are foreign and intimidating to parents of premature infants. The high levels of anxiety and stress they can produce needs to be reduced by thoughtful advice from healthcare providers (HCPs), to educate parents about their child's condition. Unfortunately time constraints often limit HCPs to only a few minutes with each baby's parents daily--only enough to convey critical information at a high level and with limited depth. Parents searching for more detailed information themselves in the literature over the web have sometimes reported disappointing experiences. We are proposing to improve parental education by patient-centric web methods leveraging the electronic patient record with internet and cell phone technologies. This can be an important informatics resource, complementing and enhancing face-to-face communication through personalization of education and advice to the parents. PMID- 17911953 TI - Conceptual model of health information ethics as a basis for computer-based instructions for electronic patient record systems. AB - A computer-based learning system called Electronic Patient Record (EPR) Laboratory has been developed for students to acquire knowledge and practical skills of EPR systems. The Laboratory is basically for self-learning. Among the subjects dealt with in the system is health information ethics. We consider this to be of the utmost importance for personnel involved in patient information handling. The variety of material on the subject has led to a problem in dealing with it in a methodical manner. In this paper, we present a conceptual model of health information ethics developed using UML to represent the semantics and the knowledge of the domain. Based on the model, we could represent the scope of health information ethics, give structure to the learning materials, and build a control mechanism for a test, fail and review cycle. We consider that the approach is applicable to other domains. PMID- 17911954 TI - PDA-based informatics strategies for tobacco use screening and smoking cessation management: a case study. AB - The purpose of this case study is to describe three incremental personal digital PDA-based informatics strategies aimed at improving screening for tobacco use and guideline-based tobacco cessation management: 1) PDA clinical log with tobacco cessation diagnoses and plan of care options, 2) PDA decision support system, and 3) PDA decision support system with infobuttons--context-specific links to the National Cancer Institute's Cancer Information Services tobacco cessation information. These strategies were implemented within the context of an evidence based advanced practice nurse curriculum at the Columbia University School of Nursing. PMID- 17911976 TI - Dry powder inhalers: which factors determine the frequency of handling errors? AB - BACKGROUND: Dry powder inhalers are often used ineffectively, resulting in a poor level of disease control. OBJECTIVES: To determine how often essential mistakes are made in the use of Aerolizer, Discus, HandiHaler and Turbuhaler and to study the effects of age, severity of airflow obstruction and previous training in inhalational technique by medical personnel on the error rate. METHODS: Two hundred and twenty-four newly referred outpatients (age 55.1 +/- 20 years) were asked how they had been acquainted with the inhaler and to demonstrate their inhalational technique. RESULTS: The inhaler-specific error rates were as follows: Aerolizer 9.1%, Discus 26.7%, HandiHaler 53.1% and Turbuhaler 34.9%. Compared to Aerolizer, the odds ratio of an ineffective inhalation was higher for HandiHaler (9.82, p < 0.01) and Turbuhaler (4.84, p < 0.05). The error rate increased with age and with the severity of airway obstruction (p < 0.01). When training had been given as opposed to no training, the odds ratio of ineffective inhalation was 0.22 (p < 0.01). If Turbuhaler is used, the estimated risks range from 9.8% in an 18-year-old patient with normal lung function and previous training to 83.2% in an 80-year-old patient with moderate or severe obstruction who had not received any training. CONCLUSIONS: Dry powder inhalers are useful in the management of younger patients with normal lung function or mild airway obstruction. In older patients with advanced chronic obstructive pulmonary disease, the risk of ineffective inhalation remains high despite training in inhalational technique. A metered-dose inhaler with a spacer might be a valuable treatment alternative in a substantial proportion of these patients. PMID- 17911977 TI - A functional disturbance in the auditory cortex related to a low serotonergic neurotransmission in women with type 2 diabetes. AB - BACKGROUND/AIMS: To determine if the slope of the amplitude/stimulus intensity function (ASF) of the N1/P2 component of the auditory evoked potential was increased in women with type 2 diabetes reflecting a low brain serotonergic activity in the auditory cortex. METHODS: In a comparative study in women with type 2 diabetes and controls, we measured free, bound and total plasma L tryptophan (L-Trp), neutral amino acids (NAA) and free fatty acids (FFA) and recorded the N1/P2 component of the auditory evoked potential. RESULTS: The diabetic patients were overweight and FFA and NAA in plasma were significantly elevated. The free, bound to albumin and total L-Trp were decreased. The values of free/total L-Trp and free/NAA ratios were significantly lower. The latencies of N1 and P2 at all intensities and the slope ASF of the N1/P2 component significantly increased. CONCLUSION: The decrease of the free fraction of L-Trp in plasma and the increase of the ASF slope of the N1/P2 component reflect a functional relationship between the brain serotonergic activity and the N1/P2 changes in the auditory cortex, suggesting a cortical impaired activity associated with anomalies of brain serotonergic neurotransmission in women with type 2 diabetes. We proposed the ASF slope together with measurement of the plasma FFT as noninvasive clinical indicators of serotonergic neurotransmission in the brain in these as well as in other types of patients. PMID- 17911978 TI - Real-time quantitative polymerase chain reaction for BCRP, MDR1, and MRP1 mRNA levels in lymphocytes and monocytes. AB - The expression levels of mRNAs for MDR1 (P-glycoprotein), multidrug resistance associated proteins (MRP1), and BCRP (breast cancer resistance protein; mitoxantrone resistance protein) were quantitatively determined in lymphocytes and monocytes. Monocytes and lymphocytes were obtained from 3 healthy male and 2 healthy female volunteers. BCRP, MDR1, and MRP1 mRNA levels were determined by real-time quantitative reverse transcriptase polymerase chain reaction. In lymphocytes, relative concentrations of mRNAs for target proteins (BCRP, MDR1, and MRP1) were 1.00 +/- 0.478, 9.67 +/- 5.53, and 0.116 +/- 0.0613 respectively, and in monocytes 1.00 +/- 0.854, 0.821 +/- 0.263, and 0.090 +/- 0.052, respectively. The MDR1 mRNA level was cell type dependent, whereas there was no difference in BCRP and MRP1 expression levels between lymphocytes and monocytes. Comparison of mRNA levels for the three major multidrug-resistant efflux pumps reveals that MDR1 is the predominant form in lymphocytes and BCRP is the predominant form in monocytes. PMID- 17911979 TI - Effects of ozone and sodium hypochlorite on caries-like lesions in dentin. AB - The hypothesis that ozone promotes remineralization of dentinal lesions was tested in vitro. Artificial caries-like lesions in dentin were treated with ozone gas, with another potent oxidizer (sodium hypochlorite, NaOCl, 10%) or with water. The specimens were then remineralized and subsequently demineralized again. Mineral content was assessed by transverse microradiography. NaOCl-treated samples showed damaged surface and, after being remineralized, demineralized significantly more than water- or ozone-treated groups. No difference was found between ozone and water groups. The exposure to ozone had thus no effect on remineralization and subsequent demineralization of remineralized dentinal lesions. PMID- 17911980 TI - Neuroimaging and APOE genotype: a systematic qualitative review. AB - Apolipoprotein E (APOE) is the major genetic risk factor for late-onset Alzheimer's disease (AD) and has also been implicated in cardiovascular disease, cognitive decline and cognitive changes in healthy ageing. The aim of this paper is to systematically review and critically assess the association between the APOE genotype and structural/functional cerebral changes as evidenced by brain imaging studies. A second aim is to determine whether these observed associations between APOE and the brain reflect changes which are consistent with the progression of AD neurodegenerative changes described in Braak stages. A search of Pubmed, Psycinfo, and Web of Science databases identified 64 articles available for qualitative review. The review found that presence of the APOE epsilon4 allele is associated with (1) hippocampal, amygdalar and entorhinal cortex atrophy, (2) increased brain atrophy, (3) increased white matter hyperintensity volumes and (4) altered cerebral blood flow and glucose metabolism patterns. It is possible that there are critical age ranges when these effects are evident and that the APOE epsilon2 genotype might present a risk. We conclude that structural brain change is associated with the APOE genotype and that it is more salient in younger ageing individuals. PMID- 17911981 TI - Vulnerability to proactive semantic interference and progression to dementia among older adults with mild cognitive impairment. AB - There is evidence that vulnerability to proactive semantic interference may be an early manifestation of early Alzheimer's disease and other neurodegenerative disorders. At present, there is a paucity of data regarding the extent to which such deficits relate to the progression of cognitive deficits and to clinically significant endpoints such as dementia. In this study, we followed 76 older adults, initially diagnosed with mild cognitive impairment, for a period of up to 3 years. Twenty-seven of these individuals (35.5%) progressed from mild cognitive impairment to dementia. An examination of baseline neuropsychological performance indicated lower baseline scores for object memory among those progressing to dementia. However, baseline Mini-Mental State Examination scores, delayed memory for passages, delayed visual memory, letter fluency, category fluency, Trails B and Block Design did not differ among study groups. In contrast, the Semantic Interference Test, a measure susceptible to vulnerability to proactive semantic interference showed the greatest baseline differentiation between those who progressed and those who did not progress to dementia. Further, scores on this measure predicted future progression to dementia with high accuracy. Vulnerability to proactive interference may be an early manifestation of an early dementing process and may have utility in predicting future progression to dementia. PMID- 17911982 TI - Dermatoporosis. The functional side of skin aging. PMID- 17911983 TI - Health-related quality of life: from health economics to bedside? PMID- 17911984 TI - Eight novel mutations of ATP2C1 identified in 17 Chinese families with Hailey Hailey disease. AB - BACKGROUND: Hailey-Hailey disease (HHD) is a rare autosomal dominantly inherited dermatosis, characterized by persistent blisters and erosions of the skin. It was recently discovered that HHD was caused by mutations in the ATP2C1 gene, a Ca2+ pump located in the Golgi apparatus. OBSERVATION: In this study, we sequenced the ATP2C1 gene from blood samples of 31 patients in 17 unrelated Chinese families and 120 healthy individuals. Eight novel mutations were identified in 9 families, including 3 insertion/deletions (nt 1464-1487/1462-1485 del, 1523 del AT, 2375 del TTGT), 3 splicing-site mutations [360(-2)a-->g, 1415(-2)a-->c, 2243(+2)t- >c], and 2 missense mutations (P307L, D648Y). CONCLUSION: Eight mutations were found in 8 unrelated families and 1 sporadic case, and these new findings have further improved our understanding of the role of ATP2C1 in HHD. PMID- 17911985 TI - Dermatoporosis: a chronic cutaneous insufficiency/fragility syndrome. Clinicopathological features, mechanisms, prevention and potential treatments. AB - BACKGROUND: Skin aging has long been considered only as a cosmetic problem. With the increase in lifespan, we are now more often experiencing a further dimension of skin aging, which is no longer only cosmetic, but also functional, in the sense that the skin has lost its protective mechanical function. Dermatoporosis is the name proposed to capture, in a holistic approach, all the aspects of this chronic cutaneous insufficiency/fragility syndrome. OBSERVATIONS: In this paper, we review the clinical aspects of dermatoporosis, its histological features and the current understanding of its etiological factors. The clinical manifestations of dermatoporosis comprise (i) morphological markers of fragility--rather trivial -such as senile purpura, stellate pseudoscars and skin atrophy, and (ii) functional expression of skin fragility resulting from minor traumas such as frequent skin laceration, delayed wound healing, nonhealing atrophic ulcers and subcutaneous bleeding with the formation of dissecting hematomas leading to large zones of necrosis. Dissecting hematomas bear significant morbidity needing hospitalization and urgent surgical procedures. Molecular mechanisms implying hyaluronate-CD44 pathways in the control and maintenance of epithelial growth and the viscoelastic properties of the extracellular matrix offer new opportunities for preventive intervention. CONCLUSION: We propose to group the different manifestations and implications of this syndrome under the umbrella term of 'dermatoporosis', because we think it will help to capture the understanding of health professionals that, as osteoporosis, 'dermatoporosis' should be prevented and treated to avoid complications. Dermatologists should be aware of this emerging syndrome and function as key players in prevention and therapy. Randomized clinical trials should demonstrate which intervention may best prevent and/or reverse dermatoporosis. PMID- 17911986 TI - Cutaneous manifestations in inflammatory bowel diseases: eight cases of psoriasis induced by anti-tumor-necrosis-factor antibody therapy. AB - BACKGROUND: Ulcerous rectocolitis and Crohn's disease are the best known forms of inflammatory bowel disease (IBD). Skin manifestations are not uncommon in IBD and may be divided into specific cutaneous signs, aspecific cutaneous signs, and cutaneous signs caused by drugs used for IBD therapy. The specific signs (fistulas, rhagades and ulcers) are the result of the diffusion of the intestinal inflammatory process into the skin. Aspecific cutaneous signs (stomatic aphthosis, erythema nodosum, pyoderma gangrenosum, Sweet's syndrome, vasculitis, bullous diseases) are quite frequently found in those suffering from IBD, but also in apparently healthy subjects, and may sometimes be the first sign of the intestinal disease. Cutaneous manifestations due to drugs vary in clinical aspect and are the direct consequence of the therapies adopted, which in IBD patients can be quite numerous: steroids, immunosuppressants, 5-aminosalicylic acid, biological agents, antibiotics. OBJECTIVE AND METHODS: Due to the frequent finding of cutaneous manifestations in patients affected by IBD, a collaboration was set up between the Dermatological Clinic of the University of Bologna and the Center for the Study of IBD of the same university hospital. The aim was to diagnose the cutaneous signs appearing during IBD and to establish their etiopathogenesis in order to assess whether they were the result of epiphenomena of the IBD or side effects of the therapies adopted. RESULTS: The cutaneous manifestations we observed can be divided into three distinct groups: signs that were specific to the basic disease, aspecific signs and finally signs attributable to the drugs used for therapy. Particular attention was given to the aspecific signs and those consequential to therapy. The aspecific cutaneous signs seen in our clinic generally reflect those reported in the literature. The cutaneous manifestations due to drugs were further divided into three groups: rosacea, acneiform dermatitis and psoriasis-like dermatitis. The most notable aspect of our series is the high number of patients presenting psoriasiform-type dermatitides with a generally widespread diffusion. CONCLUSION: We would like to draw attention to the fact that all patients with psoriasis had been undergoing treatment with drugs inhibiting tumor necrosis factor alpha (TNF-alpha) as part of IBD therapy. In all cases, the cutaneous reaction started after the third or fourth infusion of the biological drug. Anti-TNF-alpha agents have also been successfully used to treat psoriasis in the last few years. The reason for this apparently paradoxical effect of the therapy is still unclear. PMID- 17911987 TI - Low-dose X-ray depilatory treatment induces trichoblastic tumors of the scalp. AB - BACKGROUND: Depilatory radiotherapy for ringworm was largely used before antifungals were available. Patients who underwent this treatment are at high risk of developing scalp tumors or other cancers. The aim of this study was to characterize scalp tumors occurring after X-ray therapy for ringworm. METHODS: We included cases of postradiotherapy scalp tumors recorded at the Dermatology Department of the Charles Nicolle Hospital, Tunis between 1988 and 2001. We recorded clinical descriptions and all cases were resubmitted to microscopic analysis. RESULTS: Sixty-one tumors occurred in 33 men and 12 women with a mean age of 49.8 years. Radiodermatitis was present in 21% of patients. Tumors were basal cell carcinomas in 47 cases, trichoblastomas in 10 cases and trichoblastic carcinomas in 4 cases. Twelve patients had 2-5 tumors, with combinations of tumor types in 3 of them. Mean delay of onset of tumors after radiotherapy was 39.4 years in basal cell carcinoma cases, 38.3 years in trichoblastoma cases and 35.6 years in trichoblastic carcinoma cases. CONCLUSIONS: This series shows that although basal cell carcinoma is the most frequent tumor in this situation, trichoblastomas are common. We describe, for the first time, radio-induced trichoblastic carcinomas. Trichoblastic tumors have not yet been described in this context because this concept is relatively recent. PMID- 17911988 TI - Living with acne. A study of adolescents' personal experiences. AB - BACKGROUND: Acne can have important psychological repercussions. OBJECTIVES: We studied adolescents' perceptions of acne, in a nonmedical context. METHODS: Between November 2004 and January 2005, a questionnaire was presented to adolescents calling a youth helpline. It focused on perceptions of acne and, among callers with personal experience of acne, its severity and treatment. Callers were divided into those with acne, those having had acne and those never having had acne. RESULTS: The questionnaire was completed by 1,566 callers. It showed that 51% of respondents with acne did not seek medical help; acne has as big a psychological impact as some far severer diseases; many respondents with no history of acne said they were very worried that they would develop this skin disorder. CONCLUSIONS: This study underlines the major psychological impact of acne. Young people need to be informed that acne can be treated. PMID- 17911989 TI - The practical reality of using a patient-reported outcome measure in a routine dermatology clinic. AB - OBJECTIVE: To explore whether clinicians used formal quality of life (QoL) information if provided by patients completing the Dermatology Life Quality Index (DLQI) questionnaire and whether the information influenced treatment decision taking. METHODS: The DLQI was completed by adult patients attending a dermatology secondary-care clinic as they arrived at their appointment and given to the clinician. Clinicians recorded whether the DLQI information was used and whether it influenced treatment decision-making. RESULTS: A total of 417 patients attended and 268 questionnaires (64.3%) were completed. The mean DLQI score was 7 (median = 5, range = 0-30). In 64 consultations (28.8%), clinicians used the DLQI information; in 37 of these consultations (57.8%) the DLQI information influenced the clinicians' treatment decision-making. The mean DLQI score for these 37 consultations was 11.3 (median = 10.0, range = 0-29), indicating a large effect on patients' QoL. QoL discussion occurred in 85 of 98 consultations observed. The domains of QoL most frequently discussed concerned symptoms (74 consultations) and problems caused by the treatment (24 consultations). CONCLUSIONS: Clinicians may use formal QoL information if available in routine clinics. Treatment decisions may be influenced for those with high QoL impairment. Routine systematic assessment of QoL may therefore be of benefit. PMID- 17911990 TI - Pulse corticosteroid therapy for alopecia areata: study of 139 patients. AB - BACKGROUND/AIM: Recent reports of pulse corticosteroid therapy for alopecia areata (AA) show its efficacy for patients with a history of < or = 1 year but not for recalcitrant cases or alopecia totalis/universalis. The purpose of this study was to evaluate the efficacy and safety of pulse corticosteroid therapy for recent-onset AA patients. METHOD: A total of 139 severe AA patients aged >15 years were included in this study. The duration from the onset of active hair loss was within 12 months for 125 (89.9%) of those patients. RESULTS: Of the patients, 72.7% had hair loss on > 50% of their scalp area. Among the recent onset group (duration of AA < or = 6 months), 59.4% were good responders (> 75% regrowth of alopecia lesions), while 15.8% with > 6 months duration showed a good response. Recent-onset AA patients with less severe disease (< or = 50% hair loss) responded at a rate of 88.0%, but only 21.4% of recent-onset patients with 100% hair loss responded. No serious adverse effects were observed. PMID- 17911991 TI - Steroid-sparing effect of pimecrolimus cream 1% in children with severe atopic dermatitis. AB - OBJECTIVE: To investigate if pimecrolimus cream 1% reduces the need for steroids in the long-term management of severe pediatric atopic dermatitis (AD). METHODS: A total of 184 pediatric patients (aged 2-17 years) with a history of severe AD according to Rajka and Langeland were enrolled. Patients were randomized to treatment with pimecrolimus cream or vehicle cream for a 24-week period. Prednicarbate 0.25% was applied as rescue medication. RESULTS: Patients on pimecrolimus required steroids on a mean of 29% of study days, compared with 35% of patients on vehicle (p = 0.1841). On the head and neck only, the respective figures were 10 versus 19% (p = 0.0009). In patients enrolled with acute severe disease (Investigator's Global Assessment > or = 4), steroids were used on 28% of the days in the pimecrolimus group compared to 45% in the control group (p = 0.0024). On the head and neck, steroids were used on 10% of study days with pimecrolimus versus 30% with vehicle (p < 0.0001). CONCLUSION: The results indicate that the need for topical steroids on the head and neck is reduced with pimecrolimus cream 1% in the management of severe pediatric AD according to the definition of Rajka and Langeland. PMID- 17911993 TI - Quality of life impairment and psychological distress in elderly patients with psoriasis. AB - BACKGROUND/AIMS: The management of psoriasis, which has a major impact on quality of life (QoL), deserves a specific attention to elderly patients' problems and needs. Our aim was to describe the QoL of elderly patients with psoriasis. METHODS: This is a cross-sectional study on 305 psoriatic inpatients, aged > or = 65 years. QoL, measured by the Skindex-29, was compared in groups of patients based on clinical and sociodemographic characteristics, and the possible determinants of psychological distress were investigated. RESULTS: The mean age of the patients was 71 years (range: 65-93). Clinical severity tended to increase with age. Psychological distress was higher in patients > 70 years of age, while QoL showed an improvement for all the Skindex-29 scales in older age groups. Gender (women vs. men), and Skindex-29 emotions and functioning scales emerged as associated with psychological distress, after adjusting for age and clinical severity. CONCLUSION: These observations indicate that clinical severity of psoriasis increases among very elderly patients and is associated with different levels of skin-related QoL and psychological distress. It is important for dermatologists to be aware of the specific impact of psoriasis in the different subgroups of the elderly population, to meet the peculiar needs of each group. PMID- 17911992 TI - Topical retapamulin ointment, 1%, versus sodium fusidate ointment, 2%, for impetigo: a randomized, observer-blinded, noninferiority study. AB - BACKGROUND: Retapamulin is a novel pleuromutilin antibacterial developed for topical use. OBJECTIVE: To compare the efficacy and safety of retapamulin ointment, 1% (twice daily for 5 days), with sodium fusidate ointment, 2% (3 times daily for 7 days), in impetigo. METHODS: A randomized (2:1 retapamulin to sodium fusidate), observer-blinded, noninferiority, phase III study in 519 adult and pediatric (aged > or = 9 months) subjects. RESULTS: Retapamulin and sodium fusidate had comparable clinical efficacies (per-protocol population: 99.1 and 94.0%, respectively; difference: 5.1%, 95% confidence interval: 1.1-9.0%, p = 0.003; intent-to-treat population: 94.8 and 90.1%, respectively; difference: 4.7%, 95% confidence interval: -0.4 to 9.7%, p = 0.062). Bacteriological efficacies were similar. Success rates in the small numbers of sodium-fusidate-, methicillin- and mupirocin-resistant Staphylococcus aureus were good for retapamulin (9/9, 8/8 and 6/6, respectively). Both drugs were well tolerated. CONCLUSION: Retapamulin is a highly effective and convenient new treatment option for impetigo, with efficacy against isolates resistant to existing therapies. PMID- 17911994 TI - The reduced Impact of Psoriasis Questionnaire has good psychometric properties in Italian patients. AB - BACKGROUND: A recent refinement study suggested that a Rasch reduced version of the Impact of Psoriasis Questionnaire (IPSO) of 11 items most adequately assessed the psychosocial impact of US psoriasis patients. OBJECTIVE: To test whether the IPSO would also behave well in a different population that varies culturally, demographically, and in disease severity. METHODS: The psychometric properties of the IPSO, using classical test and item response theory (Rasch analysis), were assessed in 805 Italian psoriasis patients. RESULTS: Patients with more severe psoriasis reported significantly higher impact on their HRQOL (p < 0.001) and the IPSO correlated well with the Skindex-29 (r = 0.74) confirming its validity. The response distribution was adequate for all items, except item 9. The Cronbach's alphas were excellent and the high item-rest correlations confirmed its homogeneity. Principal component analysis demonstrated one dominant factor with an eigenvalue of 4.47 (items loading >0.40). Overall, the 11 IPSO items fitted the Rasch model (p = 0.07) and all items demonstrated a logical threshold order. Of the 11 items, 2 items showed significant individual misfit and only 1 item demonstrated significant differential item functioning for age but none for gender or global severity score. CONCLUSION: The 11-item IPSO is a valuable psoriasis-specific HRQOL instrument in different populations. PMID- 17911995 TI - Multiple and clustered eruptive epidermoid cysts following treatment with topical imiquimod. AB - A 61-year-old woman developed multiple and clustered eruptive epidermoid cysts at the site of treatment of a basal cell carcinoma located on her nose with imiquimod 5% cream (5 times/week for 6 weeks). Clearing was achieved after topical treatment with tretinoin 0.025% cream (1 application/day for 1 month). PMID- 17911996 TI - Dermoscopic and reflectance confocal microscope findings of trichoepithelioma. AB - BACKGROUND: Trichoepitheliomas (TE) are benign neoplasms of follicular differentiation. Solitary lesions are often confused with basal cell carcinoma (BCC). Reflectance confocal microscopy (RCM) and dermoscopy are imaging tools for in vivo, noninvasive evaluation of skin lesions. To date, there has been no description of their findings in the evaluation of TE. OBJECTIVE: Our aim is to describe the dermoscopic and RCM findings of histopathologically confirmed TE. METHODS: Four TE were evaluated, 2 each of the desmoplastic and nondesmoplastic variants. RCM was performed on 1 of the desmoplastic and both of the nondesmoplastic lesions. RESULTS: Dermoscopically, all of the lesions showed arborizing telangiectasias. The desmoplastic lesions also had an ivory-white background throughout. RCM showed oval, darker-appearing tumor islands that contained brightly refractile material, consistent with keratin horn cysts at the center, as well as parallel bundles of highly refractile dermal collagen surrounding the tumor islands. CONCLUSION: The ivory-white background throughout the lesion seen on dermoscopy may be helpful in distinguishing desmoplastic TE from BCC. The RCM findings in TE of keratin-filled cysts in tumor islands and attachment of the tumor to follicular structures have not been previously observed in BCC, and thus may also be diagnostically helpful. Further study is necessary for validation of these findings. PMID- 17911997 TI - Lack of association between CARD15 gene polymorphisms and hidradenitis suppurativa: a pilot study. PMID- 17911998 TI - SACRAL syndrome. PMID- 17911999 TI - Instability of topical ciclosporin emulsion for nail psoriasis. PMID- 17912000 TI - Prophylaxis of recurrent erythema nodosum with penicillin. PMID- 17912001 TI - The diagnostic accuracy of in vivo confocal scanning laser microscopy compared to dermoscopy of benign and malignant melanocytic lesions: a prospective study. AB - BACKGROUND: The diagnosis of melanoma at an early, curable stage is an important challenge for clinicians. Confocal scanning laser microscopy (CSLM) is a high resolution, noninvasive technology that may facilitate improved diagnostic accuracy over clinical examination. The aim of this study was to evaluate the diagnostic accuracy of CSLM compared to dermoscopy in a prospective examination of benign and malignant melanocytic lesions. METHODS: 125 patients with suspicious pigmented lesions were prospectively recruited to undergo a clinical, dermoscopic and CSLM examination. A diagnosis was made preoperatively with each technique, and the lesion was then excised and diagnosed using histopathology. RESULTS: 125 patients with 125 lesions were studied comprising 88 melanocytic nevi and 37 melanomas. Dermoscopy had a sensitivity of 89.2%, a specificity of 84.1%, a positive predictive value of 70.2% and a negative predictive value of 94.9%. CSLM was found to have a sensitivity of 97.3%, a specificity of 83.0%, a positive predictive value of 70.6% and a negative predictive value of 98.6%. No melanomas were misidentified when both techniques were used together. CONCLUSIONS: CSLM had a relatively higher sensitivity than dermoscopy; however, the specificity was similar with CSLM and dermoscopy. These results suggest that dermoscopy and CSLM are complementary. PMID- 17912002 TI - Effect of prolonged c-kit receptor inhibition by imatinib mesylate on the uterine contractility of pregnant rabbits. AB - BACKGROUND: The c-kit receptor expressed by interstitial cells in the gastrointestinal tract is crucial to their pacemaking function. The function of similar c-kit-expressing myometrial cells is unknown. METHODS: Imatinib mesylate, a specific c-kit receptor antagonist, was administered to pregnant New Zealand white rabbits (term = 31 days, n = 35) from day 27 gestation by intramuscular injection twice daily at high (50 microg/kg) or medium (10 microg/kg) dose and compared with a control group injected with vehicle only. In a second phase, two further groups received imatinib at medium or low (1 mug/kg) dose for a longer duration starting from day 18 until delivery. Three does from the latter groups as well as controls underwent myometrial biopsy under general anesthesia after spontaneous vaginal birth. Contractility was recorded by isometric tensiometry. The outcome measures were delay of parturition and in vitro contractility characteristics. RESULTS: High-dose imatinib induced early delivery when compared with the control group (28.6 vs. 30.7 days, p < 0.001). The other groups delivered at term. No effect on in vitro contractility was apparent in any of the groups. CONCLUSIONS: c-kit receptor inhibition in pregnant rabbits does not delay significantly the length of gestation or change myometrial contractility in vitro. PMID- 17912003 TI - Uterine activity after oral mifepristone administration in human pregnancies beyond 41 weeks' gestation. AB - BACKGROUND/AIM: To examine the effect of oral mifepristone on uterine activity in postterm human pregnancies. METHODS: As part of a randomized, placebo-controlled trial comparing 200 mg oral mifepristone to placebo for preinduction cervical ripening in women with well-dated pregnancies beyond 41 weeks' gestation with unfavorable cervices, uterine activity was continuously recorded with external tocodynamometry and contraction frequency tabulated. RESULTS: Ninety-seven women received mifepristone and 83 women received placebo. Uterine activity (uterine contractions/hour) was greater in the mifepristone than in the placebo group between 7 h (8.03 +/- 0.48 vs. 5.90 +/- 0.39, p = 0.001) and 24 h (8.53 +/- 0.68 vs. 6.61 +/- 0.46, p = 0.02) after dosing. CONCLUSION: Oral mifepristone administration to women with pregnancies beyond 41 weeks increases uterine activity in the absence of externally administered uterotonic agents. PMID- 17912004 TI - Spinal bone mineral density, IGF-1 and IGFBP-3 in children with cerebral palsy. AB - BACKGROUND/AIMS: Childhood cerebral palsy (CP) is associated with osteopenia and the GH-IGF axis plays an important role in bone metabolism. We studied the relationship between spinal bone mineral density (BMD) and serum IGF-1 and IGFBP 3 in children with CP. METHODS: Cross-sectional study of 30 children (9 F and 21 M, ages 4.5-15) with CP. Subjects underwent dual-energy x-ray absorptiometry scans (spinal BMD), blood tests (IGF-1, IGFBP-3, Ca, P, PTH, vitamin D, osteocalcin) and urine tests (N-telopeptide). RESULTS: Spinal BMD was decreased in children with CP (average Z-score -2.14 +/- 1.08) compared to age- and gender matched norms. IGF-1 and IGFBP-3 were also decreased compared to age-matched norms (average IGF-1 Z-score -0.74 +/- 1.2, average IGFBP-3 Z-score -0.68 +/- 1.2). All other blood and urine tests, including measures of calcium and vitamin D status, were normal. In 25 CP children with osteopenia (Z-score >-1), there was a trend towards correlation between spinal BMD Z-score and serum IGF-1 SDS score (r = 0.328, p = 0.09). IGFBP-3 Z-scores were available in 24 of these patients and had a statistically significant correlation with spinal BMD Z-score (r = 0.386, p = 0.05). CONCLUSION: Osteopenia is common in children withCP and may be associated with lower IGF-1 and IGFBP-3 levels. PMID- 17912005 TI - Reactivity of shrimp allergy-related IgE antibodies to krill tropomyosin. AB - BACKGROUND: Krill, which morphologically resembles small shrimp, represents small ocean crustaceans and has been used for human consumption in Japan and some other countries. The major allergen in crustaceans has been reported to be tropomyosin, but the allergenicity of krill tropomyosin remains uncertain. METHODS: Amino acid sequences of tropomyosin in two species of krill (Euphausia superba and E. pacifica) were deduced. Recombinant krill tropomyosins were produced in Escherichia coli using a pCold IV vector system, and the cross-reactivity of shrimp allergy-related IgE to the recombinant tropomyosins and several animal protein extracts was assessed by immunoblotting. RESULTS: The deduced amino acid sequences of the E. superba and E. pacifica tropomyosins (designated as Eup s 1 and Eup p 1, respectively) were 284 residues and showed significant homology to those of shrimp, lobster and crab tropomyosins. Shrimp allergy-related IgE reacted to approximately 38-kDa protein bands in krill (E. superba), shrimp, lobster and crab protein extracts but did not react to protein extracts from either mollusks or vertebrates. Furthermore, the IgE recognized rEup s 1 and rEup p 1 as 38-kDa protein bands, and absorption of the IgE with rEup s 1 removed IgE reactivity to recombinant tropomyosins and protein extracts from krill and shrimp. CONCLUSIONS: Krill tropomyosins included highly homologous sequences to previously reported IgE-binding epitopes in Pen a 1 (tropomyosin of Penaeus aztecus). The cross-reactivity in shrimp allergy-related IgE binding among krill, shrimp, lobster and crab tropomyosins was revealed. These observations suggest the potential allergenicity of krill tropomyosin. PMID- 17912006 TI - A comparison of inflammatory mediators released by basophils of asthmatic and control subjects in response to high-affinity IgE receptor aggregation. AB - BACKGROUND: In human blood basophils, cross-linking the high-affinity IgE receptor Fc epsilonRI with multivalent antigen activates a signaling pathway leading to secretion of inflammatory mediators and cytokine production. Basophils are known to play an important role in the pathogenesis of asthma but there has been no comprehensive examination of the effectors these cells produce. Here a study of the transcription and release of a selection of chemokines and cytokines from basophils was undertaken. METHODS: A Cartesian antibody array provided an effective method of assaying for multiple cytokines and chemokines simultaneously. Results were verified by RT-PCR and ELISA assays. This allowed the comparison of freshly prepared peripheral blood basophil responses to cross linking of the high-affinity IgE receptor, with and without preincubation with IL 3. RESULTS: Evidence that human blood basophils produce the chemokines MIP-5, eotaxin and GM-CSF was provided by antibody array and RT-PCR analyses. Preincubation with IL-3 enhanced the expression and release of IL-13, IL-8 and mRNA transcripts encoding MIP-5 and GATA2 in basophils from both asthmatic and control subjects. Leptin mRNA transcription, storage and release in basophils are described for the first time. CONCLUSIONS: Surveying cytokine and chemokines stored and released by peripheral blood basophils shows that asthmatic and control subjects share similar profiles even when their degranulation responses are distinct. Evidence is provided for the production of leptin, GM-CSF, eotaxin and MIP-5 by peripheral blood basophils. IL-3 preincubation enhances the production and release of IL-8 upon IgE receptor cross-linking. PMID- 17912007 TI - Characterization of a hypoallergenic recombinant Bet v 1 variant as a candidate for allergen-specific immunotherapy. AB - BACKGROUND: Recombinant allergens and especially their hypoallergenic variants are promising candidates for a more effective and safer specific immunotherapy. METHODS: Physicochemical and immunological characteristics of a folding variant of recombinant Bet v 1 (rBet v 1-FV) were investigated in comparison to natural Bet v 1 (nBet v 1) and the correctly folded recombinant Bet v 1 (rBet v 1-WT) by SDS-PAGE, size exclusion chromatography, multi-angle light scattering, circular dichroism, immunoblotting and enzyme allergosorbent test inhibition assay for detection of IgE reactivity and ELISA with Bet v 1-specific monoclonal antibodies. The functional IgE reactivity of the different Bet v 1 proteins was investigated using basophil activation in terms of CD203c expression and histamine release. T cell reactivity was investigated using T cell lines raised from birch pollen-allergic subjects against nBet v 1. Immunogenicity was investigated in mice. RESULTS: Physicochemical characterization revealed purity, homogeneity and monomeric properties of rBet v 1-FV. Unlike nBet v 1 and rBet v 1 WT, rBet v 1-FV showed almost no IgE binding in immunoblots. The reduction of allergenicity was further proved by IgE-binding inhibition assays, basophil activation and histamine release. T cell reactivity was completely conserved, as demonstrated by proliferation of Bet v 1-specific T cell lines with multiple epitope specificities. rBet v 1-FV showed strong immunogenicity in mice. CONCLUSIONS: Due to its reduced IgE reactivity and decreased capacity to activate basophils, but retained T cell reactivity and strong immunogenicity, rBet v 1-FV proved to be a very promising candidate for specific immunotherapy in birch pollen-allergic subjects. PMID- 17912008 TI - Primers on molecular pathways. The glycogen synthase kinase-3beta. AB - Despite tremendous scientific effort, conventional treatment approaches have had little impact on the course of pancreatic ductal adenocarcinoma. Therefore, urgency is needed to understand the molecular mechanisms underlying the development of pancreatic cancer with the hope that this will lead to preventative and treatment strategies to improve the outcome of the disease. Numerous factors contribute to progression of this disease, including constitutively active NF kappa B, which has been shown to positively influence cancer cell survival, proliferation, invasion, metastasis and chemoresistance. Recently, the cytoplasmic serine/threonine protein kinase glycogen synthase kinase-3beta (GSK-3beta) was found to regulate NF kappa B activation and the proliferation and survival of pancreatic cancer cells. Moreover, recent studies in other human malignancies have implicated GSK-3beta as a regulator of cancer cell proliferation, survival and chemoresistance through distinct mechanisms. Thus, GSK-3beta has emerged as a viable therapeutic target in the treatment of several human neoplasms. PMID- 17912009 TI - Pancreas and cystic fibrosis: the implications of increased survival in cystic fibrosis. AB - Pancreatitis affects 0.5% people with cystic fibrosis (CF) in the UK and 0.01% of the normal population. Why do some with CF get pancreatitis and some not? And does pancreatitis in neonates result in pancreatic failure with no further inflammation or risk of pancreatic cancer? Review of the literature would suggest that 85% of those with CF have pancreatic destruction as children with minimal risk of further inflammatory or neoplastic changes. Those with a functioning pancreas are at risk of developing pancreatitis. There are several case series of pancreatic cancer reported in CF patients, but overall the risk is unknown. As patients with CF and pancreatic sufficiency are living longer, further studies to assess the risk of developing pancreatic cancer in this subgroup should be considered. PMID- 17912010 TI - Cancer risk among the relatives of patients with pancreatic ductal adenocarcinoma. AB - BACKGROUND/AIMS: Pancreatic cancer is a leading cause of cancer-related death; the most consistently identified risk factors are smoking and family history. Our aims were to examine familial aggregations of pancreas and other cancers, and to determine the relative risk of the family members. METHODS: We prospectively collected data on the families of patients presenting with pancreatic ductal adenocarcinoma. Smoking habits and alcohol consumption of the probands were compared with the available statistics on the Italian population. Mortality from cancer was investigated in first-degree relatives, and age-dependent risks of dying from pancreatic cancer and other tumors were compared with background population levels. RESULTS: Data for 570 families were collected, including 9,204 relatives. Probands were 3- to 5-fold more often heavy smokers than the general population, and 9.3% of them reported a positive family history of pancreatic cancer. In first-degree relatives, only mortality from pancreatic cancer was significantly increased (relative risk at age 85 years = 2.7). Lifetime risk of dying of pancreas cancer was 4.1% for the relatives of all probands, and was 7.2% for the relatives of probands who developed disease before 60 years of age. CONCLUSIONS: The data suggest that genetic susceptibility to pancreatic cancer may be attributable, in addition to BRCA2, to moderate- to low-penetrance gene(s). PMID- 17912011 TI - Usefulness of technetium-99m hexamethylpropylene amine oxime-labeled leukocyte scintigraphy to detect pancreatic necrosis in patients with acute pancreatitis. Prospective comparison with Ranson, Glasgow and APACHE-II scores and serum C reactive protein. AB - BACKGROUND: In acute pancreatitis (AP), pancreatic necrosis (PN) is an important local complication that can be identified by means of contrast-enhanced computed tomography (CECT). Pancreatic leukocyte infiltration is a significant pathogenic event in the development of PN that can be detected by labeled leukocyte scintigraphy (LLS). The aim of this study was to evaluate the utility of LLS with technetium-99m hexamethylpropylene amine oxime ((99m)Tc-HMPAO) to detect the presence of PN in patients with AP. METHODS: Prospective cohort study of 84 patients with AP. Patients underwent LLS and the activity of images was scored on a 0-3 scale. CETC was performed to assess PN. Ranson, Glasgow and APACHE-II scores were calculated. Serum C-reactive protein (CRP) was measured. Sensitivity (Sn), specificity (Sp), positive predictive values (PPV), negative predictive values (NPV), areas under receiver operating characteristic (ROC) curves, likelihood ratios, odds ratios, analysis of variances between groups and correlation coefficients between tests were calculated. RESULTS: PN was present in 11 (13%) patients. Pancreatic labeled leukocyte uptake was present in 38 patients (45%). Sn, Sp, PPV and NPV of LLS grade 2-3 for PN diagnosis were the highest (91, 88, 53 and 98%, respectively) of all tests. Patients with LLS grade 2-3 were 71 times more likely to be at risk of PN compared to those with LLS grade 0-1. The area under ROC curve of the LLS was the largest. A significant correlation was obtained between LLS and CRP (p < 0.001). CONCLUSION: In patients with AP, LLS with (99m)Tc-HMPAO detects PN with an acceptable level of confidence and therefore could be considered an alternative technique to CECT in detecting PN. PMID- 17912012 TI - Expression of interferon-gamma- inducible protein-10 and its receptor CXCR3 in chronic pancreatitis. AB - AIM: The role of CXC chemokine, interferon-gamma-inducible protein-10 and its receptor CXCR3 in pathophysiology of chronic pancreatitis (CP) is not very clear. Hence, this study was carried out to analyze the expression of CXCL10 and CXCR3 in CP tissues. METHODS: Pancreatic tissues from 25 histopathologically graded CP cases (11 alcohol associated CP, 5 confirmed idiopathic and 9 of undefined nature) and 10 normal cases were studied. Tissues were subjected to real-time PCR, immunohistochemistry, and Western blot analysis for CXCL10 and CXCR3 expression. RESULTS: Real-time (RT)-PCR revealed increased expression of CXCL10 (13-fold) and CXCR3 (7-fold) in CP tissue. The immunohistochemistry and Western blot analysis of the same showed significant increased protein expression and correlated well with the histopathological grades. The CXCL10 was localized mainly in the cytoplasm of pancreatic acinar cells and expression increased from grade I to grade II and declined in grade III while no expression was recorded in normal. The CXCR3 was expressed strongly at the acinar cell membrane in CP as compared to normal. Further, comparative analysis by semiquantitative RT-PCR analysis was performed for other CXC/CC chemokines (CXCL9, CXCL11, CCL3, CCL4, CCL5) and receptor (CCR5) which revealed their upregulation in the diseased state. CONCLUSION: The existence of CXCL10 and CXCR3 with other CXC/CC chemokine signature in CP is suggestive of their vital role in the progression of chronic inflammation. PMID- 17912013 TI - Endoscopic ultrasound and IL-8 in pancreatic juice to diagnose chronic pancreatitis. AB - BACKGROUND/AIM: Pancreatic juice (PJ) [IL-8] has been proposed as a marker for pancreatic diseases. We compared the accuracy of PJ [IL-8] and endoscopic ultrasound (EUS) to diagnose chronic pancreatitis (CP). METHODS: 79 patients with symptoms suspicious for CP were enrolled. PJ emptied into the duodenum was collected during an upper endoscopy with IV secretin and [IL-8] was measured. CP was diagnosed when PJ [IL-8] was >20 pg/ml. CP was diagnosed at EUS when >or=4 of the 9 established criteria were present. CP was diagnosed by using composite gold standard: ERCP, histology, CT or MRI, and clinical follow-up (mean 20 months). RESULTS: 38 patients had CP, whereas 41 patients had no pancreatic disease. To diagnose CP, PJ [IL-8] was significantly less sensitive compared to EUS (47 vs. 71%), but equally accurate (71 vs. 80%) and specific (93 vs. 88%). By combining PJ [IL-8] and EUS, sensitivity and specificity significantly increased to 82% (either IL-8 or EUS positive) and 100% (both IL-8 and EUS positive). CONCLUSIONS: Both PJ [IL-8] and EUS are accurate diagnostic modalities for CP. PJ collection can be performed at the time of EUS. PJ [IL-8] and EUS are complementary with higher sensitivity and specificity when used together. PMID- 17912014 TI - In vivo radioiodide imaging and treatment of pancreatic cancer xenografts after MUC1 promoter-driven expression of the human sodium-iodide symporter. AB - INTRODUCTION AND AIMS: Mucin 1 (MUC1) is a transmembrane glycoprotein that is overexpressed in many tumor types, including breast, pancreatic, and ovarian cancer. The aim of this study was to create a construct containing sodium-iodide symporter (NIS) under the control of the 0.8-kb MUC1 promoter to infect pancreatic cancer cells both in vitro and in vivo, to investigate the potential for radioiodide imaging and ablation of this disease. METHODOLOGY: We amplified the 797-bp MUC1 promoter by two-step nested PCR. Subsequently, a replication deficient adenoviral construct was created containing the MUC1 promoter followed by the human NIS gene. Iodide uptake assays and immunofluorescence were used to confirm NIS expression and function. Pancreatic cancer xenografts in mice were infected with Ad/MUC1/NIS and then imaged and treated using radioiodide. RESULTS: A 23- and 15.5-fold increase in iodide uptake was observed in Ad/MUC1/NIS infected MUC1-positive Capan-2 and SW1990 cells with no significant increase observed in MUC1-negative Hela cells or in cells infected with the control virus. The in vivo study showed a clear image of Ad/MUC1/NIS-infected tumor xenografts using (125)I. Administration of a therapeutic dose of (131)I resulted in a regression in size to 76 +/- 15% of their original volume, whereas control tumors continued to increase in size to >200% of their original volume. CONCLUSIONS: These results show that the 0.8-kb MUC1 promoter was successfully used to drive human NIS-targeted expression in pancreatic cancer cells, and Ad/MUC1/NIS mediated radiotherapy can make pancreatic cancer xenografts in mice shrinking. This could potentially have applications for both imaging and therapy in other MUC1-positive tumors. PMID- 17912015 TI - A clinical and economic evaluation of endoscopic ultrasound for patients at risk for familial pancreatic adenocarcinoma. AB - BACKGROUND AND AIMS: Approximately 10% of pancreatic adenocarcinoma is familial. Approximately 50% of 1st-degree relatives (FDRs) have endoscopic ultrasound (EUS) findings of chronic pancreatitis. We modeled the natural history of these patients to compare 4 management strategies. METHODS: We performed a systematic review, and created a Markov model for 45-year-old male FDRs, with findings of chronic pancreatitis on screening EUS. We compared 4 strategies: doing nothing, prophylactic total pancreatectomy (PTP), annual surveillance by EUS, and annual surveillance with EUS and fine needle aspiration (EUS/FNA). Outcomes incorporated mortality, quality of life, procedural complications, and costs. RESULTS: In the Do Nothing strategy, the lifetime risk of cancer was 20%. Doing nothing provided the greatest remaining years of life, the lowest cost, and the greatest remaining quality-adjusted life years (QALYs). PTP provided the fewest remaining years of life, and the fewest remaining QALYs. Screening with EUS provided nearly identical results to PTP, and screening with EUS/FNA provided intermediate results between PTP and doing nothing. PTP provided the longest life expectancy if the lifetime risk of pancreatic cancer was at least 46%, and provided the most QALYs if the risk was at least 68%. CONCLUSIONS: FDRs from familial pancreatic cancer kindreds, who have EUS findings of chronic pancreatitis, have increased risk for cancer, but their precise risk is unknown. Without the ability to further quantify that risk, the most effective strategy is to do nothing. PMID- 17912016 TI - Early prediction of severity in acute pancreatitis using infrared spectroscopy of serum. AB - BACKGROUND: One of the main problems in the management of acute pancreatitis (AP) is the scarcity of accurate predictors of disease severity. METHODS: In a prospective design, we compared APACHE II score, C-reactive protein (CRP) level, and infrared (IR) spectral absorption of serum (wavelength 940 nm) in 167 consecutive patients with AP, 34 with predicted severe and 133 with mild form. RESULTS: The IR spectral absorption levels on admission and at 24 h after admission were significantly (p < 0.05) lower in patients with severe AP. On admission, the sensitivity was 74, 56, and 44%; the specificity was 82, 83, and 81%; the positive predictive value was 51, 45, and 37%, and the negative predictive value was 92, 88, and 85%, for IR spectroscopy, APACHE II, and CRP, respectively. At 24 h, the sensitivity, specificity, positive predictive value, and negative predictive value was 82, 74, 44, and 94%; 65, 72, 37, and 89%; 68, 73, 39, and 90%, for IR spectroscopy, CRP, and APACHE II, respectively. CONCLUSIONS: IR spectroscopy seems to be useful for early detection of severe AP and, in turn, for identifying patients requiring treatment in the intensive care unit and who can benefit from novel therapies. PMID- 17912017 TI - Hydrogen sulfide, nitric oxide and a molecular mass 66 u substance in the exhaled breath of chronic pancreatitis patients. AB - BACKGROUND/AIMS: Human exhaled breath contains many molecules either present as gases or occurring in a soluble form in the vapor of the breath. This study was designed to evaluate the substances present in the exhaled breath of chronic pancreatitis (CP) patients. SUBJECTS: Thirty-one consecutive CP patients (11 with exocrine insufficiency) and 31 healthy subjects (HS) were studied. METHODS: Ninety-eight different substances were analyzed using a mass spectrometer on a breath sample from all subjects and on each respective ambient air sample. RESULTS: H(2)S, NO and a substance having a molecular mass of 66 u (M66) were those which had significantly higher concentrations in CP patients than in HS after adjustment for the ambient air; the estimated increases attributable to the disease were 14% (p = 0.040) for H(2)S, 84% (p = 0.006) for M66 and 50% (p = 0.033) for NO, but the three volatile compounds showed poor diagnostic accuracy in differentiating CP patients from HS (AUC-ROC: 0.664, 0.715, and 0.602 for H(2)S, M66, and NO, respectively). Finally, no significant differences of H(2)S, M66, and NO were found between patients with and without alcoholic pancreatitis as well as between patients with and without pancreatic insufficiency. CONCLUSIONS: Exhaled breath analysis can rapidly and easily assess the presence of volatile compounds (H(2)S, NO and a substance having a molecular mass of 66 u) which may have properties capable of explaining, at least in part, the pathogenesis of CP. PMID- 17912018 TI - Dermal peptide delivery using colloidal carrier systems. AB - The advancement in synthetic and molecular biology techniques over the past years has resulted in the application of peptides or peptide-like drugs becoming a growing field in therapeutics. Because of the unfavorable chemical properties of peptides, it poses a challenge to find an optimized way of drug administration. The transdermal route has attracted interest as a promising way to advance the delivery of these drugs. The objective of this review is to summarize the level of research of microemulsions as colloidal carrier for dermal peptide drug delivery. The presented studies resulted in enhanced drug delivery or superior penetration profiles of peptides incorporated in microemulsions in comparison to conventional vehicles. Due to their benefits like high solubilization capacity, enhanced drug delivery, noninvasive administration or easy preparation, microemulsions offer a suitable vehicle for dermal and transdermal drug delivery. PMID- 17912019 TI - Subungual penetration of dibutyl phthalate in human fingernails. AB - Dibutyl phthalate (DBP) has a wide variety of manufacturing applications and is used in both commercial and consumer products. Results of animal reproductive toxicity and teratogenicity animal studies have not been consistent in identifying DBP as a reproductive toxicant. Expert reviews for its use in consumer products have consistently concluded that it is not a reproductive risk to consumers. Results from a subungual penetration study of 100% fluid DBP applied to human fingernails showed levels of penetration at the limits of chemical detection. Even if DBP penetrated the human fingernail, its rapid metabolism by the human body would prevent its having any toxic reproductive effects. Furthermore, DBP functions as a plasticizer in consumer products such as cosmetic nail products (nail polish, basecoats, topcoats, nail hardeners), resulting in its becoming unavailable for subungual penetration seconds after application of the cosmetic nail product since it is then trapped in the rapidly forming coating. PMID- 17912020 TI - Molecular basis for the regenerative properties of a secretion of the mollusk Cryptomphalus aspersa. AB - A screen for natural products bearing pharmacological properties has yielded a secretion of the mollusk Cryptomphalus aspersa (SCA), which possesses skin regenerative properties. In this report, we outline some of the cellular and molecular effects underlying this observation. First, we found that SCA contained antioxidant SOD and GST activities. In addition, SCA stimulated fibroblast proliferation and rearrangement of the actin cytoskeleton. Additional mechanisms involved in the regenerative effect of SCA included the stimulation of extracellular matrix assembly and the regulation of metalloproteinase activities. Together, these effects provide an array of molecular mechanisms underlying SCA induced cellular regeneration and postulate its use in regeneration of wounded tissue. PMID- 17912021 TI - Transdermal penetration of UV filters. AB - A penetration study of 2-ethylhexyl-4-methoxycinnamate (EHMC), 4-methyl benzylidenecamphor (MBC), butyl methoxydibenzoylmethane (BMBM), 2-ethylhexyl 2,4,5-trimethoxycinnamate (EHTMC) and di(2-ethylhexyl)-2,4,5 trimethoxybenzalmalonate (TMB) through baby mouse skin (Mus musculus Linn.) was carried out using a vertical Franz diffusion cell. At 4.4 mg/cm(2) coverage of UV filter on the skin, 2.98 +/- 0.38, 1.15 +/- 0.14 and 0.80 +/- 0.28% of the applied EHMC, MBC and BMBM were detected in the receptor fluid at 24 h after application. Penetrations of UV filter in an ethanolic solution and lotion forms were comparable. EHTMC and TMB showed insignificant penetration across the baby mouse skins. Baby mouse skins kept at 4, -20 and -80 degrees C gave similar EHMC penetration results. Penetrations of EHMC, BMBM, EHTMC and TMB across human epidermis were carried out upon 5 volunteers using the suction blister technique. The results also confirmed the significant penetrations of EHMC and BMBM and the insignificant penetrations of EHTMC and TMB. PMID- 17912023 TI - Autophagy is required for dietary restriction-mediated life span extension in C. elegans. AB - Dietary restriction extends life span in diverse species including Caenorhabditis elegans. However, the downstream cellular targets regulated by dietary restriction are largely unknown. Autophagy, an evolutionary conserved lysosomal degradation pathway, is induced under starvation conditions and regulates life span in insulin signaling C. elegans mutants. We now report that two essential autophagy genes (bec-1 and Ce-atg7) are required for the longevity phenotype of the C. elegans dietary restriction mutant (eat-2(ad1113) animals. Thus, we propose that autophagy mediates the effect, not only of insulin signaling, but also of dietary restriction on the regulation of C. elegans life span. Since autophagy and longevity control are highly conserved from C. elegans to mammals, a similar role for autophagy in dietary restriction-mediated life span extension may also exist in mammals. PMID- 17912024 TI - HDAC6 at the intersection of autophagy, the ubiquitin-proteasome system and neurodegeneration. AB - The two major intracellular catabolic pathways, the ubiquitin-proteasome system (UPS) and macroautophagy (autophagy), have each been implicated as playing roles in neurodegenerative proteinopathies. We have investigated the relationship between the UPS and autophagy using Drosophila models of neurodegenerative diseases. We identified histone deacetylase 6 (HDAC6) as a genetic modifier of polyglutamine-induced neurodegeneration and determined that its mechanism of action is autophagy-dependent. The ability of HDAC6 to suppress degeneration has been extended to additional neurodegenerative disease models, including a fly model expressing pathological Abeta fragments, presented here, but is not a universal modifier of degenerative phenotypes. Importantly, HDAC6 was also found to suppress degeneration associated with proteasome mutations in an autophagy dependent manner, revealing a compensatory relationship between these two degradation pathways. Our findings indicate that HDAC6 facilitates degradation of potentially noxious protein substrates, contributing vitally to the neuroprotective role of autophagy. PMID- 17912025 TI - Aberrant membranes and double-membrane structures accumulate in the axons of Atg5 null Purkinje cells before neuronal death. AB - Autophagy (macroautophagy) is an evolutionally conserved process by which cytoplasmic proteins and organelles are surrounded by unique double membranes and are subsequently degraded upon fusion with lysosomes. Many autophagy-related genes (Atg) have been identified in yeast; a ubiquitin-like Atg12-Atg5 system is also essential for the elongation of the isolation membrane in mammalian cells. Nevertheless, the regulation of autophagy in neurons remains largely unknown. In this study, we crossed conditional knockout mice Atg5(flox/flox) with pcp2-Cre transgenic mice, which express Cre recombinase through a Purkinje cell-specific promoter, pcp2. In Atg5(flox/flox); pcp2-Cre mice, the Atg5 gene was excised as early as postnatal day 6; Purkinje cells started to degenerate after approximately 8 weeks, and the animals showed an ataxic gait from around 10 months. Initially, however, the Purkinje cells showed axonal swelling around its terminals from as early as 4 weeks after birth. An electron microscopic analysis revealed the accumulation of autophagosome-like double-membrane structures in the swollen regions, together with numerous membranous organelles, such as tubular or sheet-like smooth endoplasmic reticulum and vesicles. These results suggest that Atg5 plays important roles in the maintenance of axon morphology and membrane structures, and its loss of function leads to the swelling of axons, followed by progressive neurodegeneration in mammalian neurons. PMID- 17912026 TI - A naturally occurring truncated beta3 integrin in tumor cells: native anti integrin involved in tumor cell motility. AB - Alternatively spliced integrins may play an important role in integrin mediated tumor cell adhesion, spreading, and migration. Here we report in human tumor cells a naturally occurring alternatively spliced variant of the beta3 integrin [i.e., truncated (tr) beta3] that lacked a cytoplasmic and a transmembrane domain. The presence of trbeta3 was demonstrated at the mRNA level by RT-PCR, cloning, and sequencing; at the protein level by immunohistochemistry and Western Blotting. The alternately spliced beta3 integrin was detected in human prostate carcinomas, breast carcinomas, and melanoma cells. Expression in vivo was confirmed by immunohistochemistry with an antibody to trbeta3 that does not recognize wild type beta3. Tumor cells secreted this protein and deposited it on the extracellular matrix. Secreted trbeta3 inhibited adhesion of melanoma and prostate cancer cells to fibronectin and vitronectin, which was partially reversed by adsorption of trbeta3 from the media. Confocal microscopy and time lapse live cell microscopy demonstrated that trbeta3 distributed to the trailing edge of migrating cells, which may represent an alternative cell detachment mechanism in these cells. Results suggest that trbeta3 may act as an anti integrin and play a crucial role in cell migration, which is an important process in tumor invasion and metastasis. PMID- 17912027 TI - The pilot study of anti-tumor effects versus immunosuppression of Staphylococcal Enterotoxin C. AB - Superantigens tremendously activate T lymphocytes by recognizing the particular region on TCR Vbeta, by which cytotoxic T cells can be well armed to kill tumor cells. However, the obstacle exists in the fact that immunosuppression is induced adversely. Staphylococcal enterotoxins (SEs) are pyrogenic superantigens who invoke T lymphocytes cytotoxicity at a very low dosage where their endotoxic activity diminishes. Despite that the elaborate mechanisms are largely unknown, tumoricidal capacity of SEA and SEB has been well studied. In this study, we devoted our attention to evaluate Staphylococcal Enterotoxin C (SEC) regarding its tumoricidal activity versus immunosuppression. We proved with flow cytometry that SEC treatment on C57 mice resulted in a boost of the differentiation of T lymphocytes into CD4(+), CD8(+) subpopulations. In vitro, SEC causes increased IFNgamma release from human PBMC. Furthermore, in coculture SEC-treated human PBMC led to more death of cancer cell lines from a variety of origins. Systemic SEC treatment in mouse and rabbit models significantly decreases tumor growth. In tumor-bearing rabbits, tumor necrosis and strong infiltration of lymphocytes into tumor tissue were observed; the rabbits also benefit with less metastasic cancer cells in the lung. In the meantime, the induced cell immune responses, both T cell differentiation and PBMC IFNgamma release, declined as SEC concentration rose. Tumor growth data obtained from animal models are in accordance with the changes in immunity, in which tumor growth ceased to respond to high dosage SEC as it did to lower dosage. These observations on SEC investigation, particularly in aspect of dosage-related immunosuppression, are of significance to SEC therapeutic potential to cancer. Molecular mechanism underlying these findings warrants further intensive investigation. PMID- 17912028 TI - Influence of apoptosis (BCL2, FAS), cell cycle (CCND1) and growth factor (EGF, EGFR) genetic polymorphisms on survival outcome: an exploratory study in squamous cell esophageal cancer. AB - The study aimed at investigating whether genetic polymorphisms in BCL2, FAS, CCND1, EGF and EGFR genes influence the outcome of patients of esophageal squamous cell cancer treated with radiotherapy, with or without chemotherapy. Sixty nine histologically confirmed, previously untreated, patients with a squamous cell esophageal cancer were inducted into this study. Genotyping of BCL2 (ala43thr), FAS (A-670G), CCND1 (G870A), EGF (+61A/G) and EGFR (G497A) polymorphisms were determined using the polymerase chain reaction followed by restriction fragment length polymorphism methodology. Genotyped data was analyzed using univariate and multivariate logistic regression statistical tests for predicting the survival outcome. Genotypes of BCL2, FAS, CCND1 and EGFR polymorphisms independently did not influence outcome significantly. However, patients with EGF +61AG genotype had median survival of 25.5 months (95% CI = 5.2 45.5), whereas those with EGF +61GG genotype had survival of only 3.7 months (95% CI = 0.0-9.8, p = 0.006). In univariate cox-regression analysis, interaction of genotypes EGF+61GG*radiotherapy tumor dose (< or =50 Gy) and EGF +61GG *upper third tumor location showed high hazard of death, 6.6 (95% CI = 2.0-21.5, p = 0.002) and 26.8 (95% CI = 3.7-194.2, p = 0.001) while EGF+61AG*middle third tumor location had reduced hazard 0.20 (95%CI = 0.06-0.60, p = 0.004). The pilot study suggests that EGF +61AG and +61GG genotypes may predict clinical outcome in esophageal cancer patients treated with radiotherapy with or without chemotherapy. EGF +61AG genotype was associated with improved survival, however +61GG genotype adversely affected the outcome in patients particularly with upper third location of tumor and lower dose (< or =50) of radiotherapy. PMID- 17912030 TI - Copy-number variants in patients with a strong family history of pancreatic cancer. AB - Copy-number variants such as germ-line deletions and amplifications are associated with inherited genetic disorders including familial cancer. The gene or genes responsible for the majority of familial clustering of pancreatic cancer have not been identified. We used representational oligonucleotide microarray analysis (ROMA) to characterize germ-line copy number variants in 60 cancer patients from 57 familial pancreatic cancer kindreds. Fifty-seven of the 60 patients had pancreatic cancer and three had nonpancreatic cancers (breast, ovary, ovary). A familial pancreatic cancer kindred was defined as a kindred in which at least two first-degree relatives have been diagnosed with pancreatic cancer. Copy-number variants identified in 607 individuals without pancreatic cancer were excluded from further analysis. A total of 56 unique genomic regions with copy-number variants not present in controls were identified, including 31 amplifications and 25 deletions. Two deleted regions were observed in two different patients, and one in three patients. The germ-line amplifications had a mean size of 662 Kb, a median size of 379 Kb (range 8.2 Kb to 2.5 Mb) and included 425 known genes. Examples of genes included in the germ-line amplifications include the MAFK, JunD and BIRC6 genes. The germ-line deletions had a mean size of 375Kb, a median size 151 Kb (range 0.4 Kb to 2.3 Mb) and included 81 known genes. In multivariate analysis controlling for region size, deletions were 90% less likely to involve a gene than were duplications (p < 0.01). Examples of genes included in the germ-line deletions include the FHIT, PDZRN3 and ANKRD3 genes. Selected deletions and amplifications were confirmed using real-time PCR, including a germ-line amplification on chromosome 19. These genetic copy-number variants define potential candidate loci for the familial pancreatic cancer gene. PMID- 17912029 TI - Evaluation of GATA-4 and GATA-5 methylation profiles in human pancreatic cancers indicate promoter methylation patterns distinct from other human tumor types. AB - The GATA-4 and GATA-5 transcription factors are increasingly recognized as playing a role in carcinogenesis of human tumors derived of endodermal and mesodermal origin. The pancreas is derived from endodermal tissues suggesting GATA-4 and GATA-5 gene methylation may play a critical role in the biology of human pancreatic cancer as well. We investigated GATA-4 and -5 by methylation specific PCR (MSP) in normal and neoplastic pancreatic tissues, including isogenic xenografts or cultured cell lines derived from the coexistent primary cancer and/or metastases in patients with pancreatic carcinoma. The relationship of promoter methylation was correlated with mRNA expression for each gene, and methylation patterns were correlated with known clinicopathologic features of patients. GATA-4 demonstrated a significantly lower methylation frequency than GATA-5 in low passage pancreatic cancer xenografts or cell lines (1/34 versus 21/34, p < 0.001). GATA-4 and -5 were also evaluated in microdissected samples of normal duct epithelium and cancer from pancreas cancer tissues which confirmed infrequent GATA-4 methylation in pancreatic cancers as well as in normal duct epithelium. GATA-4 was frequently overexpressed at the mRNA level with 27 of 30 (90%) pancreatic cancers showing >5.0-fold overexpression compared to normal duct epithelial cells. By contrast, high frequency methylation of GATA-5 was confirmed in pancreatic cancers tissues, but was rarely methylated in normal duct epithelium, indicating hypermethylation of this gene during pancreatic cancer development. GATA-5 mRNA expression did not correlate with its promoter hypermethylation, and treatment with the demethylating agent 5-aza-2' deoxycytidine only partially restored mRNA expression suggesting additional regulatory mechanisms of GATA-5 expression. The presence of GATA-5 methylation showed a trend towards worse long-term survival (14.0 +/- 9.2 months versus 19.5 +/- 3.9 months, p = 0.06). While hypermethylation of GATA-5 seems to be a universal feature among human tumors, infrequent methylation of GATA-4, and its corresponding overexpression, appears unique to pancreatic cancer from other tumor types reported thus far. PMID- 17912031 TI - Hyperspectral image analysis of live cells in various cell cycle stages. AB - In this study we have explored the use of hyperspectral imaging (HSI) to determine the cell cycle status of live cells in culture. Live cancer cell lines in culture were either synchronized by release from nocodazole or arrested in various cell-cycle phases with serum starvation (G1), aphidicolin (S), or nocodazole (G2/M). The live cells were then stained with the fluorescent DNA binding dyes Heochst 33342 or DCO along with propidium iodide or MTR. Samples were examined using fluorescence microscopy and entire spectral emission profiles were acquired for each sample using a PARISS HSI system. Classified spectra were incorporated into spectral libraries. All spectra acquired from each sample were correlated with library spectra to a user-determined confidence threshold, generating unique spectral signatures for each sample. Examination of these spectral signatures revealed that all cell cycle phases could be objectively differentiated. Ongoing studies employing other viable cell fluorescent dyes, and dyes in combination may provide more robust spectral signatures defining the status and condition of living cells. PMID- 17912032 TI - Effects of low confluency, serum starvation and hypoxia on the side population of cancer cell lines. AB - The cancer stem cell theory describes a small subset of cancer cells that have the ability to initiate and drive the growth of a tumor. The niche refers to the environmental factors and the surrounding cells within which the tumor develops. The exact relationship between cancer stem cells and the tumor niche is not known. However, using side population analysis by flow cytometry, it is possible to analyze the relationship between environmental stresses and putative cancer stem cells. The side population is a subpopulation of cells that efflux Hoechst 33342 and has been previously shown to be enriched for cancer stem cells. Using this technique, we characterized the response of side population cells to low confluency, serum starvation and hypoxia using three different human cancer cell lines. We found that these stresses, characteristic of the tumor niche enrich the side population of DLD1, SW480 and MCF7 cancer cell lines, thus possibly predisposing the tumor to a more malignant phenotype. PMID- 17912033 TI - Replication stress, defective S-phase checkpoint and increased death in Plk2 deficient human cancer cells. AB - We previously reported that the Polo-like Kinase 2 gene (Plk2/Snk) is a direct target for transcriptional regulation by p53 and that silencing Plk2 sensitizes cancer cells to Taxol-induced apoptosis. Our goals have been to better understand why Plk2 is regulated by p53 and how Plk2 signals protection from cell death through checkpoint activation. We found that following knock-down of Plk2 in wild type p53 expressing H460 human non-small cell lung cancer cells there was a significant increase in cell death observed in aphidicolin-treated cells and a further increase after release from aphidicolin-block. The highest levels of cell death were observed when Plk2-deficient cells were released from both aphidicolin and etoposide treatment. These results suggested that a defective S-phase checkpoint may contribute to enhanced sensitivity of Plk2-deficient cells to replication stress. Consistent with this hypothesis, we observed higher levels of Serine 139 H2AX phosphorylation in Plk2-deficient as compared to control cells before and after aphidicolin treatment indicating that there is more DNA damage when Plk2 is depleted. We also observed higher levels of Chk1 protein in Plk2 deficient cells that were associated with reduced levels of Serine 317 phosphorylated Chk1. In aphidicolin-treated cells, there were lower levels of Serine 317-phosphorylated Chk1 when Plk2 was knocked-down. Plk2 was demonstrated to interact with Chk2, Chk1, Serine 317-phosphorylated Chk1 and p53. Thus, increased cell death observed after aphidicolin treatment and release in Plk2 deficient cells may result from both higher levels of replication stress-induced DNA damage and a dysfunctional S-phase checkpoint. PMID- 17912034 TI - Morphogens and the control of cell proliferation and patterning in the spinal cord. AB - The development of animal embryos depends on accurate coordination of the growth and specification of precursor cells. Morphogens, extracellular signals that act at a distance to control cell fate, are crucial in the patterning of embryonic tissues. One of the most extensively studied examples of a morphogen patterned tissue is the developing vertebrate spinal cord. The distribution of distinct neuronal subtypes along the dorsoventral (DV) axis of the spinal cord is determined by counteracting gradients of long-range signals. Wnt and BMP signals promote dorsal identities, while Shh signaling induces ventral identities. Simultaneous to their specification, neural progenitors proliferate, facilitating the growth of the neural tube. In this review we discuss evidence indicating that the signals governing progenitor specification also control proliferation and survival of progenitor cells. Moreover, evidence of reciprocal transcriptional interactions and cross-talk between the signaling pathways has emerged from recent studies. Together these studies suggest ways in which patterning and growth may be coordinated in the spinal cord. One level of interaction is an inhibitory regulation of repressor forms of the transcription factor Gli3 - generated in the absence of Shh - on beta-catenin activity, the transcription factor activated by Wnt signaling. This interaction may also be relevant in other tissues and situations in which the two signaling pathways are known to participate. PMID- 17912035 TI - AIF-mediated programmed necrosis: a highly regulated way to die. AB - Historically, two main forms of cell death have been distinguished: apoptosis and necrosis. Apoptosis was initially considered as the only physiological and programmed form of cell death. This type of death is recurrently associated with caspases, a family of cysteine proteases activated in apoptotic conditions. However, it is now widely recognized that programmed cell death (PCD) can also occur in the complete absence of caspase activation. The existence of non-caspase PCD pathways was corroborated by the discovery of caspase-independent executioners, such as the mitochondrial protein Apoptosis-Inducing Factor (AIF). Necrosis has often been viewed as an accidental and uncontrolled cell death process. Nevertheless, increasing evidence shows that, like apoptosis, necrosis could be a highly orchestrated type of PCD. Indeed, apoptosis and necrosis present more similarities than it has been originally thought. Here, we summarize the different classifications of PCD and the current knowledge of a necrotic PCD pathway mediated by AIF: alkylating DNA-damage mediated death. We also outline the molecular mechanisms controlling this form of PCD and discuss their potential relevance in physiological and pathological settings. These emerging data on the molecular mechanisms regulating programmed necrosis may certainly have potent therapeutic consequences in treating both apoptotic-resistant tumors and degenerating adult neurons. PMID- 17912036 TI - Inhibition of paclitaxel-induced proteasome activation influences paclitaxel cytotoxicity in breast cancer cells in a sequence-dependent manner. AB - Although the anti-tumour effects of paclitaxel result mainly from mitotic arrest, recent evidences suggest alternative mechanisms of cytotoxicity. Cell cycle, cell death, and gene expression assays were used to understand the molecular mechanisms of paclitaxel cytotoxicity in breast cancer cells. G(2)/M cell cycle arrest and cell death coincided with the regulation of genes involved in cell death, cell cycle control, microtubule-based processes, oxidative stress, and ubiquitin-proteasome system. Induction of proteasome genes was also correlated with an accumulation of protein for proteasome subunits. Furthermore, a schedule dependent regulation of paclitaxel-induced cytotoxicity was observed after combining paclitaxel and the proteasome inhibitor MG132. Proteasome inhibition after paclitaxel exposure induced the highest rate of growth inhibition and apoptosis, with no effect on mitotic arrest. These findings give support to clinical combinations of taxanes with proteasome inhibitors, outlining the importance of considering the sequence when designing such regimens. PMID- 17912037 TI - E2F-dependent induction of p14ARF during cell cycle re-entry in human T cells. AB - The ARF protein, encoded by alternate exon usage within the CDKN2A locus, provides a link between the retinoblastoma (pRb) and p53 tumor suppressor pathways. Agents that disable pRb or otherwise impinge on the E2F family of transcription factors induce expression of ARF, resulting in stabilization of p53 and activation of p53-regulated genes. However, in some cell types ARF is not induced upon cell cycle re-entry, as expected of a conventional E2F target gene, leading to the suggestion that the ARF promoter only responds to supra physiological or aberrant levels of E2F. These properties have recently been attributed to a variant E2F binding site but attempts to map specific response elements within the ARF promoter have generally yielded confusing answers. Here we show that in IL2-dependent T-lymphocytes, ARF expression is induced as cells progress from G(0) into S phase, in parallel with other bona fide E2F target genes. This is accompanied by increased association of E2F1 with the endogenous ARF promoter. Our findings suggest that the ability of ARF to register normal proliferative cues depends on the levels of E2F generated in different settings and argue against the idea that it reacts exclusively to oncogenic signals. PMID- 17912038 TI - Potassium-chloride cotransporter mediates cell cycle progression and proliferation of human corneal epithelial cells. AB - Epidermal growth factor (EGF)-induced proliferation of corneal epithelial cells contributes to its renewal, which maintains the protective and refractive properties of the cornea. This study characterized in human corneal epithelial cells (HCEC) the role of the potassium-chloride cotransporter (KCC) in mediating (1) EGF-induced mitogen-activated protein kinase (MAPK) pathway activation; (2) increases in cell cycle progression; and (3) proliferation. The KCC inhibitor [(dihydroindenyl)oxy] alkanoic acid (DIOA) and KCC activator N-ethylmaleimide (NEM), suppressed and enhanced EGF-induced p44/42MAPK activation, respectively. Such selective modulation was mirrored by corresponding changes in cell proliferation and shifts in cell cycle distribution. DIOA induced a 20% increase in G(0)/G(1)-phase cell population, whereas NEM induced a 22% increase in the proportion of cells in the G(2)/M-phase and accelerated the transition from G(0)/G(1)-phase to the S-phase. Associated with these changes, KCC1 content in a plasma membrane enriched fraction increased by 300%. Alterations in regulatory volume capacity were associated with corresponding changes in both KCC1 membrane content and activity. These results indicate that EGF-induced increases in KCC1 activity and content modulate cell volume changes required for (1) activation of the p44/42MAPK signaling pathway, (2) cell cycle progression, and (3) increases in cell proliferation. PMID- 17912039 TI - p53 is nuclear and functional in both undifferentiated and differentiated neuroblastoma. AB - Aberrant cytoplasmic sequestration has been reported as an alternative mechanism of p53 inactivation to mutation in neuroblastoma. We hypothesized that p53 localization and function in neuroblastoma is related to differentiation status. Eighty-two untreated and 24 paired pre and post-chemotherapy neuroblastomas were studied by immunocytochemistry for p53, p21(WAF1), BAX, Bcl2 and Ki67. Predominantly nuclear p53 was detected in undifferentiated neuroblastoma, and both nuclear and cytoplasmic p53 in differentiating neuroblastoma. The nuclear p53 labeling index (LI) correlated with the Ki67 LI (r = 0.51, p <0.001), and weakly with p21(WAF1) (r = 0.37), but not with BAX or Bcl2. There was a significant reduction in p53, p21(WAF1) and Ki67 LI after chemotherapy (p < 0.01), an increase in BAX (p <0.05), but no change in Bcl2. p53 localization and function were examined in two p53 wild-type undifferentiated and 9-cis retinoic acid differentiated neuroblastoma cell lines. Using immunocytochemistry, immunofluorescence and cell fractionation, p53 was found to be predominantly nuclear in both undifferentiated and differentiated cells. Following irradiation, there was upregulation of p53, p21(WAF1) and MDM2, but less induced PARP and caspase 3 cleavage in differentiated cells, suggesting intact p53 transcriptional function, but resistance to apoptosis. p53 function in undifferentiated and differentiated cells was confirmed by upregulation of p21(WAF1) and MDM2 following Nutlin-3 treatment. In conclusion, p53 is predominantly nuclear and functional in neuroblastoma regardless of differentiation status. PMID- 17912040 TI - Imaging of nucleolar dynamics during the cell cycle of cancer cells in live mice. AB - The synthesis and assembly of ribosomal subunits take place in the nucleolus. The nucleolus forms in the nucleus around the repeated ribosomal gene clusters and undergoes cyclic changes during the cell cycle. Although the nucleolus is easily visualized by light microscopy of cells in vitro, the nucleolus has not been imaged in cells in vivo. We report here development of a mouse model to visualize the nucleolus cycle of cancer cells in live mice. HT-1080 human fibrosarcoma cells were labeled in the nucleus with histone H2B-GFP and with retroviral RFP in the cytoplasm. The nucleolus was visualized by contrast to the fluorescence of GFP expressed in the nucleus. HT-1080 dual-color cells were seeded on the surface of a skin-flap of nude mice. The inside surface of the skin-flap was directly imaged with a laser scanning microscope 24 hours after seeding. The nucleoli of the cancer cells were clearly imaged in real-time. The appearance of the nucleoli changed dramatically during the cell cycle. During mitosis, the nucleolus disappeared. After mitosis, the nucleoli decreased in number and increased in size. The nucleolus appears to have a major role in cell cycle regulation. Nucleolar imaging could be used for more precise determination of cancer-cell position in the cell cycle in vivo. PMID- 17912042 TI - Sex does matter: comments on the prevalence of male-only investigations of drug effects on rodent behaviour. AB - Despite abundant evidence of sex differences in the effects of drugs on nonsexual behaviour in rats and mice, most researchers continue to investigate male animals exclusively. This was evident from a survey of all relevant research reports published during the period February 2005-September 2006 (inclusive) in recent issues of five representative behavioural pharmacological journals. Reasons for excluding female animals from most studies are discussed along with attempts to justify the use of either male or female animals only, and the value of including both sexes (especially when a drug effect is poorly understood). Although there are other factors that can influence the effects of drugs, such as strain, age and social density, the sex of experimental animals is the easiest to control and thus is well suited to inclusion in pharmacological investigations. It is accordingly suggested that, as has been recommended many times in the past, animals' sex should play a more important part in future research than is still currently the case. PMID- 17912041 TI - Loss of CABLES1, a cyclin-dependent kinase-interacting protein that inhibits cell cycle progression, results in germline expansion at the expense of oocyte quality in adult female mice. AB - Recent studies have shown that cell cycle inhibitors encoded by the Ink4a gene locus constrain the self-renewing activity of adult stem cells of the hematopoietic and nervous systems. Here we report that knockout (KO) of the Cables1 [cyclin-dependent kinase (CDK)-5 and ABL enzyme substrate 1] cell cycle regulatory gene in mice has minimal to no effect on hematopoietic stem cell (HSC) dynamics. However, female Cables1-null mice exhibit a significant expansion of germ cell (oocyte) numbers throughout adulthood. This is accompanied by a dramatic elevation in the number of atretic immature oocytes within the ovaries and an increase in the incidence of degenerating oocytes retrieved following superovulation of CABLES1-deficient females. These outcomes are not observed in mice lacking p16INK4a alone or both p16INK4a and p19ARF. These data support recent reports that adult female mice can generate new oocytes and follicles but the enhancement of postnatal oogenesis by Cables1 KO appears offset by a reduction in oocyte quality, as reflected by increased elimination of these additional germ cells via apoptosis. This work also reveals cell lineage specificity with respect to the role that specific CDK-interacting proteins play in restraining the activity of adult germline versus somatic stem cells. PMID- 17912043 TI - Acute anorectic response to cannabinoid CB1 receptor antagonist/inverse agonist AM 251 in rats: indirect behavioural mediation. AB - Despite a large and consistent literature on the suppressant effects of cannabinoid CB1 receptor antagonists/inverse agonists (e.g. rimonabant, AM 251) on food intake and weight gain in rodents, surprisingly little is known about the behavioural selectivity of such effects. In this study, ethological scoring was used to characterize the acute behavioural effects of the rimonabant analogue AM 251 (1.5 and 3.0 mg/kg, intraperitoneally) in nondeprived male rats during a 1-h test with palatable mash. Data were also collected on daily weight gain and on retest food intake 7 days after dosing. Results showed that the higher dose of AM 251 significantly inhibited mash consumption (32% decrease relative to vehicle control), reduced time spent feeding during the test and suppressed body weight gain over the 48-h period that followed acute dosing. No effects on mash consumption were observed when the animals were retested drug-free 1 week after drug treatment. Detailed video analysis of the test sessions showed that, over the dose range tested, AM 251 did not significantly interfere with the vast majority of noningestive behaviours. Both doses of the compound, however, significantly increased the incidence of and the time spent on scratching, whereas the higher dose additionally increased both the number and duration of grooming episodes. The latter effect in particular disrupted the normal structure of behaviour (behavioural satiety sequence) with atypically high levels of grooming displacing feeding during the middle part of the test session. Overall, the behavioural profile of AM 251 in a free-feeding context is very similar to (but approximately two-fold less potent than) that recently reported for the parent molecule, rimonabant. Together, these data strongly suggest that the acute anorectic response to CB1 receptor antagonists/inverse agonists is indirectly mediated via major alterations to other components of the behavioural repertoire. PMID- 17912044 TI - The monoamine oxidase inhibitor phenelzine enhances the discriminative stimulus effect of nicotine in rats. AB - In addition to delivering nicotine, tobacco smoke also inhibits monoamine oxidase (MAO). Although MAO inhibitors (MAOIs) can increase nicotine self-administration in rodents, the effects of MAOIs on the discriminative stimulus effect of nicotine are not known. This study examined the effects of three MAOIs (phenelzine, clorgyline and pargyline) with varying selectivity for MAOA and MAOB in the nicotine drug discrimination procedure in rats. Adult male Sprague-Dawley rats were trained to discriminate nicotine (0.3 mg/kg, subcutaneously) from saline in a standard, two-lever food-reinforced operant task. Once the discrimination was acquired, the ability of each MAOI to substitute for or alter the discriminative stimulus effect of nicotine was determined. In substitution tests, nicotine (0.03-0.3 mg/kg) produced full, dose-dependent substitution. Although the selective MAOA inhibitor clorgyline (3-56 mg/kg) and the selective MAOB inhibitor pargyline (3-56 mg/kg) did not elicit any nicotine-appropriate responding, partial substitution was obtained with the nonselective MAO inhibitor phenelzine (1-17 mg/kg). Phenelzine (10 mg/kg) also enhanced the discriminative stimulus effect of a low dose of nicotine (0.056 mg/kg) and prolonged the time course effect of the nicotine-training dose. These findings indicate that concomitant inhibition of MAOA and MAOB can enhance the discriminative stimulus effect of nicotine in rats. PMID- 17912045 TI - Changes in sensitivity of response distributions to changing reinforcement ratios during exposure to ephedrine, caffeine, and ephedrine-caffeine combinations. AB - Changes in the sensitivity of response distributions to changes in reward distribution (reinforcer distribution sensitivity) were examined when rats were exposed to low and moderate doses of caffeine, ephedrine, and caffeine-ephedrine combinations. The data show significant decreases in sensitivity in response distributions to changes in reward schedule values during exposure to caffeine and ephedrine/caffeine combinations, whereas ephedrine alone resulted in overmatching comparable with baseline and NaCl conditions. Rats treated either with 3.0-mg/kg or 10.0-mg/kg doses of caffeine and all combinations of ephedrine at doses of 1.8 or 5.6 mg/kg with caffeine at 3.0 or 10.0 mg/kg showed reduced sensitivity in response distributions to differences in reinforcement schedule ratios. In contrast, when rats were exposed to ephedrine at 1.8 or 5.6 mg/kg, they maintained or increased the degree of overmatching. Although reinforcer distribution sensitivity was altered, drug exposure did not significantly affect the absolute rates of responding. Bias varied after exposure to caffeine, ephedrine, and their combinations, but not systematically. Finally, whereas the estimates of goodness of fit (r2) to the matching equation showed some decreases during drug exposure, these were neither statistically significant nor correlated with drug dose. These results suggest differential effects of ephedrine and caffeine on the sensitivity of response distributions to changes in reinforcement ratio distributions, with deleterious effects of caffeine and ephedrine/caffeine combinations. PMID- 17912046 TI - Antidepressant-like effect of tramadol in the unpredictable chronic mild stress procedure: possible involvement of the noradrenergic system. AB - Tramadol, which inhibits the reuptake of noradrenaline and serotonin, is effective in animal models of depression. Its antidepressant-like effects may be mediated mainly by the noradrenergic system. This study investigated the role of the noradrenergic system in the antidepressant-like effects of tramadol and desipramine in the unpredictable chronic mild stress model. We assessed the involvement of beta-adrenoreceptors, particularly beta2-receptors in the activity of these drugs. In addition, we measured the level of noradrenaline and its metabolite 3-methoxy-4-hydroxy-phenylglycol (MHPG) in the locus coeruleus, hypothalamus, hippocampus and cerebellum in stressed mice. Unpredictable chronic mild stress induced a degradation of coat state and decreased grooming behaviour in the splash test, which was reversed by the chronic administration of tramadol (20 mg/kg) and desipramine (10 mg/kg). The nonselective beta-adrenoreceptor antagonist propranolol (5 mg/kg, intraperitoneally) and the selective beta2 receptor antagonist ICI 118,551 (2 mg/kg, intraperitoneally) reversed the antidepressant-like effects of tramadol and desipramine. Moreover, chronic tramadol and desipramine treatment increased the level of noradrenaline (NA) and MHPG in the locus coeruleus but not in the cerebellum, whereas only MHPG level was increased in the hypothalamus. Tramadol, however, increased the levels of MHPG and NA in the hippocampus, whereas desipramine only increased NA level. These data support the view that the noradrenergic system plays an important role in the antidepressant-like action of tramadol. PMID- 17912047 TI - Probability discounting among cigarette smokers and nonsmokers: molecular analysis discerns group differences. AB - Despite numerous studies documenting differences in temporal discounting among cigarette smokers and nonsmokers, studies of probability discounting have had mixed results. This study sought to clarify these results with a molecular analysis of data obtained from a study of probability discounting. Thirty heavy cigarette smokers and 29 nonsmokers were compared using a probability discounting procedure. When these data were fit to a hyperbolic model of discounting, nonstatistically significant group difference between smokers and nonsmokers were observed in the predicted direction. A molecular analysis of indifference points indicated that statistically significant differences between groups were obscured by a possible floor effect. Specifically, indifference points obtained from high probabilities were lower for heavy cigarette smokers relative to nonsmokers, whereas no difference was observed with indifference points obtained from low probabilities. This molecular analysis suggests that the prior inconsistent results are due to floor effects resulting from greater inclusion of low probabilities. Rate of smoking may also be an important factor, with differences more likely observed when heavy smokers are enrolled. PMID- 17912048 TI - Female and male rats in late adolescence differ from adults in amphetamine induced locomotor activity, but not in conditioned place preference for amphetamine. AB - Rodent models display differences in drug-induced behaviour between prepubertal/young adolescents and adults that parallel developmental differences in people; however, little is known as to when the transition to 'adultlike' behaviour occurs. We investigated the differences in locomotor and reward responses to amphetamines in male and female rats in late adolescence and compared them with corresponding adult responses. Long-Evans rats were tested for locomotor activity and conditioned place preference (CPP) for amphetamine (0.25, 0.5 or 1.0 mg/kg), beginning at 45 or 69 days of age. Adolescent female rats moved less to the first injection of amphetamine compared with adult female rats irrespective of dose, whereas adolescent male rats did not differ from adults. Adolescent female rats significantly increased locomotor activity in response to subsequent injections of amphetamine at all three doses, whereas such sensitization was only found at the highest dose for adult female and male rats. No effect of repeated injections at any dose was observed in adolescent male rats. No age differences were observed in CPP, but female rats showed greater CPP during the dioestrous than during the oestrous phase of the cycle. These data suggest that differences in neural systems underlying some behavioural responses to amphetamine continue to mature postpubertally into late adolescence in a sex specific manner. PMID- 17912049 TI - Behavioral response to emotional stress in rabbits: role of serotonin and serotonin2A receptors. AB - Exposure to a novel environment is a stressor which modulates behavior, increases stress hormones and enhances the release of several neurotransmitters including serotonin (5-HT). Exposing rabbits to a novel environment significantly increases head-bob behavior but fails to alter either grooming or wet dog shakes compared with those observed in the home-cage. The goal of this study was to determine the role of 5-HT and its receptors in mediating novelty-elicited head-bob behavior. Reduction of central 5-HT levels after treatment with the serotonergic neurotoxin 5,7-DHT significantly decreased novelty-elicited head bobs by 40% compared with those in sham-lesioned rabbits, indicating that 5-HT mediates, in part, this behavior. Additionally, pretreatment with the 5-HT1A partial agonist and clinically used anxiolytic buspirone also significantly attenuated novelty elicited head bobs. Pretreatment with the selective 5-HT2A antagonist M 100,907 significantly reduced novel environment-elicited head bobs by 40%. Furthermore, agonist-induced reduction of cortical 5-HT2A receptor density resulted in a significant 40% reduction in the number of head bobs elicited by the novel environment. These data demonstrate that rabbit head-bob behavior, an index of the response to novelty stress, is mediated, in part, by 5-HT activation of 5 HT2A receptors. PMID- 17912050 TI - Ontogenetic difference in ethanol reinforcing properties: the role of the opioid system. AB - Previous data indicate that ethanol intoxication (3 g/kg, intragastric) on postnatal day (PD) 7 and 8 increases ethanol acceptance, but on PD 10 and 11 generates an aversion in infant rats. We investigated the participation of the opioid system in these effects. Subcutaneous administration of naloxone (1 or 10 mg/kg) followed by ethanol intoxication on PD 7 and 8 prevented the increased ethanol intake effect observed in the younger pups, but when ethanol intoxication occurred on PD 10 and 11, naloxone treatment did not affect the aversion observed at this age. An aversion to ethanol was evidenced in the younger pups administered ethanol and naloxone, but only when exposed to ethanol odor during ethanol intoxication. Results indicate that the increased ethanol acceptance induced by ethanol intoxication in the younger pups is mediated by the opioid system, and that ethanol may also induce conditioned aversions at this early age. PMID- 17912051 TI - Chronic forced swim stress inhibits ultra-low dose morphine-induced hyperalgesia in rats. AB - Ultra-low doses of morphine (UL-morphine) induce hyperalgesia, which is assumed to be mediated by stimulatory G proteins (G(alphas)) signaling pathway. G(alphas) pathway inhibition and chronic stress both attenuate development of tolerance to analgesic effect of morphine. This study evaluated the effect of chronic stress on UL-morphine-induced hyperalgesia to find out if chronic stress interacts with the G(alphas) signaling pathway. Repeated daily forced swim stress was applied to induce chronic stress. UL-morphine (1 microg/kg, intraperitoneal)-induced hyperalgesia was assessed using the tail-flick test on day 6, in male rats that during days 1-5 received different treatments of swim stress, dexamethasone, swim stress following adrenalectomy (ADX) or swim stress after sham operation. Chronic stress by itself induced hyperalgesia in control and sham-operated rats but inhibited UL-morphine-induced hyperalgesia. In ADX animals, chronic stress did not produce hyperalgesia and could not inhibit UL-morphine-induced hyperalgesia. Chronic dexamethasone produced hyperalgesia but did not change the UL-morphine induced hyperalgesia. Inhibition of UL-morphine hyperalgesia by chronic stress suggests that chronic stress interacts with the G(alphas) signaling pathway, which is responsible for UL-morphine-induced hyperalgesia. The absence of this effect in the ADX-rats or after repetitive dexamethasone administration demonstrates that hypothalamic-pituitary-adrenal (HPA) axis activation is necessary for controlling UL-morphine-induced hyperalgesia. Finally, the interaction of stress with the G(alphas) signaling pathway may provide an explanation for the inhibitory effect of stress on development of tolerance to the analgesic effect of morphine. PMID- 17912052 TI - Effects of AM1346, a high-affinity CB1 receptor selective anandamide analog, on open-field behavior in rats. AB - AM1346 is a cannabinoid receptor type 1 (CB1R) anandamide analog [alkoxyacid amide of N-eicosa-(5Z, 8Z, 11Z, 14Z)-tetraenylamine] with high affinity and selectivity for the CB1 vs. CB2 receptor [Ki (CB1)=1.5 nmol/l; Ki (CB2)=152 nmol/l]. The present study characterized the effects of AM1346 (5.6-18 mg/kg) and its interaction with the CB1R antagonist/inverse agonist SR141716 (1-5.6 mg/kg) on open-field behaviors of rats. AM1346 reduced ambulation (horizontal activity), rearing (vertical activity) and increased the degree of circling and the latency to leave the central area of the open-field arena. AM1346 also tended to reduce defecation and to increase vocalization in a dose-dependent manner. In pretreatment studies, SR141716 completely blocked the effects of AM1346 on circling and latency and partially antagonized the effects of 18 mg/kg AM1346 on ambulation and rearing. SR141716 also tended to decrease AM1346-induced vocalization in a dose-dependent manner. Earlier studies have shown that SR141716, given alone, can increase grooming behavior and, as well, produces dose related increases in scratching. In the present studies, these effects were attenuated in a dose-related manner by AM1346. The present profile of behavioral effects for AM1346 is consistent with its designation as a CB1R agonist. When combined with drug discrimination data (surmountable antagonism of effects of SR141716 by Delta(9)-THC and AM1346 but not by methanandamide, i.e. AM356), these data indicate that the anandamide analog AM1346 may be more behaviorally similar to cannabinoids like Delta(9)-THC than to other anandamide-based molecules such as methanandamide. PMID- 17912053 TI - Behavioural and pharmacological specificity of the effects of drugs on punished schedule-induced polydipsia. AB - Wistar rats were exposed to a multiple fixed-time 30-s food delivery schedule, with an on/off tone signalling the two components. Animals were matched in accordance with the levels of schedule-induced polydipsia. Drinking was then punished in one of the components: half of the rats received lick-dependent 10-s signalled delays and the other half lick-dependent electric shocks. The intensities of the shocks were adjusted to reduce behaviour by the same amount as the delays in food presentation. Unpunished components were used as yoked-control conditions, by presenting delays or shocks independently of the animals' behaviour. D-Amphetamine (0.3-2.0 mg/kg) and cocaine (1.0-10.0 mg/kg) dose dependently increased (although only slightly) and then decreased schedule induced polydipsia punished with lick-dependent delays in food presentation, a result not observed in control conditions or when the behaviour was suppressed by lick-dependent electric shocks. Diazepam (1.0-17.0 mg/kg) and pentobarbital (3.0 17.0 mg/kg) dose-dependently increased and then decreased only the schedule induced drinking punished with lick-dependent shocks. Buspirone (0.1-1.0 mg/kg) and morphine (2.0-5.6 mg/kg) showed either no specific effects or further suppressed schedule-induced drinking. Results of these and previous experiments suggest that the antipunishment effects of drugs depend not only on the precise nature of the drug, but also on the manner in which the behaviour is maintained. PMID- 17912054 TI - Influence of nicotinic receptor modulators on CB2 cannabinoid receptor agonist (JWH133)-induced antinociception in mice. AB - Delta9-tetrahydrocannabinol is the active component in cannabis and has long been associated with pain relief. This effect is believed to be mediated through central and peripheral CB1 and peripheral CB2 receptors. We have explored the possible antinociceptive effect of a CB2 receptor agonist, JWH133, using the formalin test in mice. The drug was administered by the intracerebroventricular and intraperitoneal routes. Although no antinociceptive effect was observed after intracerebroventricular administration of JWH133, when the drug was administered by the intraperitoneal route, it produced an analgesic effect. The influence of nicotinic cholinergic receptor modulators, nicotine and mecamylamine, on antinociceptive effect of JWH133 was also studied. Nicotine increased and mecamylamine decreased the antinociceptive effect of JWH133. It is concluded that JWH133-induced analgesia is influenced by nicotinic cholinergic receptor activity. PMID- 17912055 TI - Repeated treatment with the NMDA antagonist MK-801 disrupts reconsolidation of memory for amphetamine-conditioned place preference. AB - Long-lasting drug-associated memories can contribute to relapse; therefore these memories must be inactivated to enable sustainable success in addiction therapy. As drug associations are usually acquired over several conditioning events, we assume that an effective treatment should be repeatedly applied to achieve persistent effects. In this study, we examine whether 10 repeated memory reactivation tests followed by systemic N-methyl-D-aspartate receptor antagonist MK-801 (0.1 mg/kg) administrations can disrupt memory reconsolidation in rats, leading to a reduction of well-established amphetamine-conditioned place preference (CPP). We found that immediate (but not 60-min delayed) administration of MK-801 after the tests reduced amphetamine-CPP expression after at least four treatments. These effects were specific to CPP expression as no MK-801-induced change in locomotion was observed during all tests. We discuss these results as being caused by MK-801 disrupting memory reconsolidation and we propose the applied repeated-treatment regimen as a new therapeutic research strategy to persistently disrupt drug-associated memories. PMID- 17912056 TI - C.T. Brighton/ABJS Workshop on Computer-assisted Orthopaedic Surgery. PMID- 17912057 TI - THE CLASSIC: On a method of investigating the deep ganglia and tracts of the central nervous system (cerebellum). Br Med J 1906:1799-1800. PMID- 17912058 TI - Computer evaluation of kinematics of anterior cruciate ligament reconstructions. AB - Current clinical and instrumented outcome measurements of knee instability lack accuracy, especially when multiplanar instability is considered. The aim of our cadaveric study was to describe the kinematics in the intact, double bundle, and anteromedial bundle reconstructed anterior cruciate ligament knee by applying a protocol for computer-assisted evaluation of knee kinematics. An optical navigation system was used to acquire knee motion (n = 5) during clinical evaluations by tracking markers rigidly attached to the bones. The protocol included acquisition of anteroposterior translations and internal-external rotations and evaluation of three clinical knee laxity tests (anterior drawer, manual, and instrumented Lachman). Our anteroposterior translation data showed the double-bundle technique and anteromedial bundle technique could restore anteroposterior stability comparable to the intact state. For internal-external laxity, the double-bundle technique demonstrated overcorrection at 15 degrees, 60 degrees, 75 degrees, and 90 degrees. The anterior drawer and manual Lachman knee laxity tests showed improved stability for the double-bundle compared to the anteromedial bundle technique. This pilot study suggests the computation of knee laxity with a high precision method might be a step toward a more precise kinematic test of knee stability for evaluating different reconstruction methods. PMID- 17912059 TI - Acute treatment of inversion ankle sprains: immobilization versus functional treatment. PMID- 17912060 TI - Lateralization of interictal EEG findings. AB - Several reports indicate that interictal epileptiform discharges (IED) may be more likely to occur over the left cerebral hemisphere than over the right. The objective of our study was to determine the frequency and type of IED on routine and multihour EEGs in a tertiary epilepsy center to estimate the frequency of left-sided versus right-sided IED and to determine interictal spike distribution pattern differences between adult and pediatric epilepsy patients. The current study retrospectively reviewed 31,207 EEGs (25,793 routine EEGs and 5414 multihour EEGs) recorded on 24,003 patients during the period from 1993 to 2003. All EEGs were read according to a systematic EEG classification system. Every patient was considered only once by including the first abnormal EEG. Regional unilateral or bilateral IEDs were recorded in 1707 patients (7%). Regional unilateral or bilateral slow was recorded in 2297 patients (9.6%). Left-sided regional IED were seen in 828 patients and accounted for 58% of all unilateral IED. Left-sided slow was seen in 1389 patients and accounted for 65% of all unilateral slow. Lateralization of slow was due to intermittent slow, whereas continuous slow involved both hemispheres equally. There was no lateralization difference in benign focal epileptiform discharges of childhood. Lateralization shows a tendency toward greater left-sided lateralization of interictal findings with aging. Benign focal epileptiform discharges were only seen under the age of 20 years old. Regional IEDs were seen in approximately 7% of patients and slowing occurs in 10% of patients. Both abnormalities were seen more frequently in the left hemisphere. Age adjusted analysis of the data revealed that this left-sided predominance was mildly increased in adults as compared with pediatric patients. PMID- 17912061 TI - Electroencephalographic characteristics of patients infected with west nile virus. AB - SUMMARY: : The EEG is helpful in the evaluation of patients with altered mental status and can provide clues for the underlying cause of certain entities. There are few descriptions of the EEG findings in patients with West Nile virus (WNV) infection. We describe the clinical presentation as well as the electroencephalographic findings in five patients with WNV encephalitis. Review of the records of all cases of WNV infection seen at the Mayo Clinic from 1999 to 2003, in which an EEG had been performed. Five patients with WNV encephalitis, in whom an EEG was performed, were found. All the patients had altered mental status and the EEG showed moderate to severe degrees of generalized slowing. Three of the patients had triphasic waves on the EEG. The patients did not have any metabolic disorder or electrolyte abnormalities that could account for these findings. EEG findings in WNV are consistent with an encephalopathic pattern and show varying degrees of generalized slow wave abnormalities and in some cases triphasic waves. PMID- 17912062 TI - Short-term outpatient EEG-video monitoring with induction in a veterans administration population. AB - The gold standard for diagnosis of psychogenic non-epileptic seizures (PNES) is EEG-video monitoring. EEG-video monitoring is usually prolonged and inpatient, but the availability of this procedure for veterans is limited. This study thought to evaluate the yield of short-term outpatient EEG-video monitoring for the diagnosis of PNES in a V.A. population. We reviewed the data on all short term outpatient EEG-video monitoring performed at our V.A. hospital over a 2-year period. Short-term EEG-video monitoring was performed with induction according to a published protocol []. Briefly, induction is performed without a placebo, using hyperventilation, photic stimulation, and verbal suggestion. This was performed on patients in whom there was a clinical suspicion of PNES on clinical grounds. A total of 52 short-term EEG-video monitoring sessions were performed. Of those, 40 patients (77%) were men. In 35 patients (67%) the procedure recorded the habitual episode and resulted in a clear diagnosis of PNES. The procedure was inconclusive in 17 patients (33%), either because a non-habitual event was induced (7 patients, 14%), or no event was recorded (10 patients, 19%). The yield of EEG video monitoring with induction in a (predominantly male) V.A. population is high, and comparable to a non-V.A. population. PMID- 17912063 TI - Temporal stability of auditory event-related potentials in healthy individuals and patients with traumatic brain injury. AB - Historically, cognitive event-related potentials (ERPs) have received limited acceptance for clinical use due to lack of evidence for their reliability. However, recent advances in computer technology and artifact rejection methods have greatly enhanced the fidelity of ERP measurements. The present study examined the test-retest reliability of ERP measurement by using current data processing methods. We assessed the temporal stability of the most commonly used ERP paradigm, auditory pure-tone "odd ball" detection, and compared it with other commonly used clinical measures reported in the literature. Auditory ERPs were collected in 19 healthy subjects and 7 patients with traumatic brain injury at two time points, 2 days to 2 months apart. Test-retest reliability was calculated for four ERP components: N1, MMN (mismatch negativity), P3, and N4. In healthy subjects, temporal stabilities of these four commonly studied ERP components' amplitude measurements were moderate to high, with intraclass correlations ranging from 0.6 to 0.8. In contrast, in patients with traumatic brain injury, ERPs were stable only for the N1 component (intraclass correlation = 0.7). PMID- 17912064 TI - The effects of increasing stimulus complexity in event-related potentials and reaction time testing: clinical applications in evaluating patients with traumatic brain injury. AB - This study compared the effectiveness of P300 event related potentials (ERPs) and reaction time (RT) in discriminating patients with traumatic brain injury (TBI) from healthy control subjects. In particular, we examined how the use of more complex, ecologically relevant stimuli may affect the clinical utility of these tasks. We also evaluated how length of posttraumatic amnesia (PTA) and loss of consciousness (LOC) related to P300 and RT measures in our patient sample. There were 22 subjects (11 patients with TBI and 11 age-matched healthy control subjects). Four stimulus detection procedures were used: two using simple, conventional stimuli (auditory tone discrimination, AT; visual color discrimination, VC), and two using complex, ecologically relevant stimuli in the auditory and visual modalities (auditory word category discrimination, AWC; visual facial affect discrimination, VFA). Our results showed that RT measures were more effective in identifying TBI patients when complex stimuli were used (AWC and VFA). On the other hand, ERP measures were more effective in identifying TBI patients when simple stimuli were used (AT and VC). We also found a remarkably high correlation between duration of PTA and P300 amplitude. PMID- 17912065 TI - Diagnostic value of event-related evoked potentials N200 and P300 subcomponents in early diagnosis of Alzheimer's disease and mild cognitive impairment. AB - Event-related potentials (ERPs) have a large application in the evaluation of cognitive processes, particularly in Alzheimer's disease (AD). The aim of the present study was to evaluate the clinical relevance of event-related evoked potentials (N2 and P3 subcomponents) in early diagnosis of AD and mild cognitive impairment (MCI). We prospectively studied 60 subjects. They all underwent the following investigations: neurologic and neuropsychological examination; functional evaluation, i.e., ERPs; cerebral imagery (morphologic and functional). Subjects were classified into 3 groups: group 1: 30 dementia of Alzheimer type (NINCDS-ADRDA, DSM-IV criteria); group 2: 20 MCI; and group 3: 10 control subjects. ERPs were significantly different between the groups (AD, MCI, control subjects), with a marked increase of P3 latencies, particularly when compared with N2 latencies (P < 0.0001). Furthermore, sensitivity was 87% to 95% for the differentiation of AD patients from MCI and control subjects, using prolonged P3 latencies (specificity, 90% to 95%), whereas using N2 prolonged latencies, sensitivity was 70% to 75% (specificity, 70% to 90%). Moreover, in the MCI group, N2 latencies strongly discriminated MCI from control subjects, with 90% sensitivity and 70% specificity and correctly categorized 80% of MCI subjects against 73% for P3. The abnormalities of N2 and P3 components may be linked, in AD and MCI, to an alteration of automatic and controlled attention processing. PMID- 17912066 TI - Validation of a rational malingering test using evoked potentials. AB - Malingering is easy to define, difficult to detect, and very costly for any health care system. The structured interview of reported symptoms (SIRS) was constructed using rational strategies to detect malingering in patients endorsing psychotic symptoms. This study validated the SIRS using evoked potentials. Nineteen patients meeting DSM-IV criteria for schizophrenia and 15 healthy controls completed an oddball and paired click protocol. Severity of psychotic symptoms was documented using the Scale for the Assessment of Negative Symptoms and the Scale for the Assessment of Positive Symptoms. The patient group was divided by probability of malingering according to the SIRS. Patients with a high probability of malingering had significantly greater P3 amplitude (P = 0.006, t test) and more P50 suppression (P = 0.044, t-test) than patients with a low probability of malingering. No significant difference in P3 amplitude or P50 suppression was found between the patients with a high probability of malingering and the healthy controls. This study provides empirical support for the validity of the SIRS with evidence that is independent of patient report. PMID- 17912067 TI - Does the recruitment of excitation and inhibition in the motor cortex differ? AB - The level of excitability within the motor cortex can be described as a balance between excitation and inhibition, but it is unknown how well both processes correlate. To address this question, the authors measured motor cortical excitability and inhibition in healthy human subjects, comparing the recruitment of motor evoked potentials (MEPs) and the duration of the cortical silent period (CSP) after transcranial magnetic stimulation (TMS). Single-pulse "focal" TMS was applied at intensities varying between 90% and 200% of motor thresholds to the right motor cortex of 15 healthy volunteers. The peak-to peak size of MEP responses and the duration of the CSP were measured in small hand muscles. Stimulus-response (S-R) curves were constructed by plotting the MEP size and CSP duration against stimulus intensities. The absolute duration of CSP and the size MEPs correlated significantly and to a similar extent with stimulus intensity (r = 0.60 and 0.53, respectively). The slope of the MEP-S-R was steeper compared with CSP-S-R, particularly at low stimulation intensities. CSP duration saturated earlier and CSP-S-Rs were shifted upwards at a given stimulus intensity compared with MEP-S-Rs. The findings suggest that recruitment of inhibition and excitation within the sensorimotor cortex correlate. However, inhibitory effects are recruited at lower intensities and saturate earlier than excitation. PMID- 17912069 TI - Sympathetic skin response and axon counting in carpal tunnel syndrome. PMID- 17912070 TI - Fear factor, freeze out, finger pointing, and the initiative for the profession, version 07.09. PMID- 17912071 TI - Radioprotection of craniofacial bone growth. AB - In this review, the potential of pharmacologic therapy for prevention of radiation-induced bone growth inhibition is discussed. Significant radioprotection using the radioprotector Amifostine has been achieved in animal models of radiation-induced retardation of long and craniofacial bone growth. Moreover, radioprotection in vitro has been achieved in a number of cell lines, including osteoblast-like, endothelial, and fibroblastic. This evidence may support future clinical investigations of radioprotector Amifostine or similar substances for radioprotection of the growing craniofacial skeleton. PMID- 17912072 TI - Radiation-induced craniofacial bone growth disturbances. AB - Multimodality treatment, including radiotherapy, chemotherapy, and surgery, is required for the management of head and neck cancer in pediatric patients. Despite the modern advances in radiation dosing and targeting techniques, the radiation damage to the growing craniofacial skeleton in children remains a significant clinical problem. The first part of this review summarizes the clinical effects of radiotherapy on craniofacial bone growth in children. Experimental evidence on therapeutic radiation effects on bone growth in in vivo and in vitro models is reviewed. The second part of this review focuses on prevention of radiation-induced craniofacial bone growth inhibition using radioprotective agents. PMID- 17912073 TI - Multicenter study of wound healing in neurofibromatosis and neurofibroma. AB - Based on clinical experience, the senior author has become convinced that wounds produced to correct the deformities of patients with neurofibromatosis (NF-1) have produced remarkably good scars, the interesting feature being that progression to keloid or hypertrophic scar is rare. The other point noted was that this situation did not change, no matter the patient's race or skin color. There have been few reports describing or discussing this hypothesis. The purpose of this study was to investigate whether wounds produced in the patients with NF 1 produce keloid or hypertrophic scars. The patients with solitary neurofibroma were also included in this study; these were compared with the NF-1 group. This was conducted as a multicenter study. Patients with neurofibromatosis/solitary neurofibroma, who were operated on from 1990 to 2000, were evaluated by reviewing their medical charts and photographs retrospectively. The patients were treated in centers from five different countries. The analysis was undertaken based on the following points: 1) age and sex at surgery; 2) race of the patients; 3) past and family histories of hypertrophic scar and keloid; 4) surgical site(s); 5) diagnosis, NF1 or solitary neurofibroma; 6) surgical complications; 7) number of reoperations to manage the complications; 8) adjuvant therapy for the tumor; 9) depth of the tumors; and 10) incidence of malignant degeneration. A total of 101 cases with neurofibromatosis or solitary neurofibroma was analyzed. The age at surgery ranged from 1 year 6 months to 74 years; sex ratio was 47 males and 54 females. The racial distribution of the patients was 13 white, 13 black, 3 Hispanic, and 58 Asian. There was no past or family history of hypertrophic scar or keloid. The surgical sites were head and neck in 70 cases, trunk in 20 cases, upper extremities in 22 cases, and lower extremities in 20 cases. The clinical diagnosis was NF-1 in 57 cases, solitary neurofibroma in 35 cases, plexiform neurofibroma in four cases, and no distinct clinical diagnosis in five cases. There were no other types of neurofibromatosis. Hematoma and white wide scar were the main postoperative complications found in six cases of NF-1. Infection was also noted in four cases. However, no patient developed hypertrophic scar or keloid in the neurofibromatosis group, whereas two cases showed hypertrophic scar in the solitary neurofibroma group. The outcome showed that the patients with NF 1 and plexiform neurofibroma, no matter the racial group, produce good scars without keloid or hypertrophic changes, whereas solitary neurofibroma has a potential to cause hypertrophic scar. PMID- 17912075 TI - Report of a radiation-induced leiomyosarcoma arising from the posterior neck. AB - Radiation-induced leiomyosarcoma of the head and neck region has been very seldom described. Herein, we report a 48-year-old male patient who developed leiomyosarcoma in his posterior neck region, which was previously radiated due to a cerebellar astrocytoma. PMID- 17912074 TI - Surgical therapy for osteonecotic lesions of the jaws in patients in therapy with bisphosphonates. AB - OBJECTIVE: Surgical therapy to improve the symptoms and the lesions in osteonecrosis (ON) of the jaws in patients in therapy with bisphosphonates. DESIGN: to evaluate the patient's therapeutic protocols, performance status, and factors promoting ON to prevent surgical failure. MATERIALS AND METHODS: 18 patients affected by osteonecrotic lesions of the jaws associated to BF, were treated by surgery. RESULTS: The results were recorded after 6 months. All the patients showed improvement of symptoms, in particular the pain. In addition, all the patients referred to a sensation of fresh and clean mouth, the disappearance of fetor ex ore, and a healthy mouth. CONCLUSIONS: The management and the resolution of BF osteonecrotic lesions is arguable and complex because in most cases, the patients are affected by oncologic disease when the better approach is prevention, but when the ON lesion is clear, surgery can improve the symptoms and in some cases, it can be resolute. To prevent surgical failure, it can be useful to evaluate the patient's therapeutic protocols, performance status, and factors promoting ON. PMID- 17912077 TI - Endoscopic-assisted excision of esthesioneuroblastoma. AB - The purpose of this article is to report a case of esthesioneuroblastoma involving the bilateral paranasal sinuses, which was excised using an endoscopic assisted transfacial approach. A patient presented with nasal swelling and left sided nasal obstruction, epistaxis, and diplopia. Examination revealed broadening of the nasal dorsum with a fleshy pink mass in both nasal cavities. Computed tomographic scan showed a mass involving the nasal cavity and paranasal sinuses on both sides. The tumor was diagnosed as group C esthesioneuroblastoma. The mass was excised by bilateral medial maxillectomy and bilateral frontoethmoidectomy. Using a 0 degrees endoscope, the attachment of the tumor to the cribriform plate was identified and resected using a motordrill. On Waroff staining, Hispathology slides suggested esthesioneuroblastoma. The patient was asymptomatic for 1 year, following which he developed infection of the nasal cavity for which he had no form of treatment. He subsequently developed maggots in the nasal cavity after which he died. An endoscopic resection of the cribriform plate from the nasal cavity without a formal craniofacial resection can be safely performed with oncologic safety. PMID- 17912076 TI - Nonmelanoma skin tumors involving the craniofacial region: our 22 years of experience. AB - Skin tumors comprise the largest group of malignancies of the head. Despite the accessibility of such lesions, the treatment of neglected, far advanced cancers, many of which have extended deeply into the facial bones and skull, is often required. The key to the cure of malignant tumors of the head is an accurate diagnosis and evaluation of the margins of an excised tumor. Reconstructive surgery of the head after resection of tumors requires a complete understanding of the anatomy of this region. From January 1986 to December of 2005, 31 patients underwent reconstructive surgery for nonmelanoma skin tumors involving the craniofacial region. Preoperative evaluation of the patients was performed in all cases. The results were estimated from the oncologic and functional point of view. The reconstruction, which was performed, included local, regional, and free flaps. In our series, the 5-year disease-free survival rate was 87%. The primary goal of surgical treatment of skin tumors with invasion of craniofacial bone structure is three-dimensional tumor resection with histologically clear margins. This goal has to be balanced, however, with an acceptable functional and aesthetic result. Resections are planned according to pathologic considerations rather than according to the anatomy involved. PMID- 17912078 TI - Maspin expression in oral squamous cell carcinoma. AB - Maspin (mammary serine protease inhibitor) is a member of the serpin superfamily of protease inhibitors and it has a role as a tumor suppressor. Maspin has been reported to be important in processes relevant to tumor growth and metastasis such as cell invasion, angiogenesis, and apoptosis. A high expression of maspin was correlated with better rates of survival and absence of nodal metastases in head and neck squamous cell carcinoma. In contrast, some studies have shown that maspin overexpression is correlated with a poor prognosis in pancreatic and ovarian cancers and in lung adenocarcinoma. The aim of this study was an immunohistochemical evaluation of the maspin expression in oral squamous cell carcinoma and thus 89 patients were evaluated. Maspin expression in oral squamous cell carcinoma was significantly associated with the tumor differentiation grade (chi test: P = 0.0318) and the lymph node status (chi test: P < 0.005), but not with the tumor stage (chi test: P = 0.666). Metastatic involvement of lymph nodes was observed more frequently in maspin-negative cases than in tumors with more than 5% of positive cells (P = 0.0024). The present results confirm that maspin expression predicts a better prognosis in oral squamous cell carcinoma and that maspin probably plays a role in tumor progression. PMID- 17912079 TI - An in vitro model of radiation-induced craniofacial bone growth inhibition. AB - Radiation-induced craniofacial bone growth inhibition is a consequence of therapeutic radiation in the survivors of pediatric head and neck cancer. Previously, the infant rabbit orbitozygomatic complex (OZC) was established as a reliable animal model. The purpose of this study was to develop a cell culture model from the rabbit OZC to study the effects of radiation in the craniofacial skeleton. Infant (7-week-old) New Zealand white rabbits were used in this study. Periostea from both OZC were harvested in sterile conditions, introduced into cell culture by way of sequential digestion, and subcultured at confluence. Cultures were analyzed for cellular proliferation (methylthiazoletetrazolium assay), alkaline phosphatase activity, collagen type I expression, and mineralization. Electron microscopy was performed to reveal the in vitro ultrastructure. Subsequently, rabbits were irradiated with sham or 15 Gy radiation, and cell cultures were developed and analyzed for cell numbers. Cell cultures, grown from OZC periostea, expressed osteoblast-like phenotype, with high alkaline phosphatase activity, collagen type 1 expression, and mineralization in an osteogenic environment. Electron microscopy confirmed the characteristic ultrastructural features of osteogenesis in vitro. Finally, significantly (P < 0.01) fewer cells were obtained from animals treated with 15 Gy radiation compared with those from control animals.A primary cell culture with osteoblast-like cellular phenotype was developed from infant rabbit OZC periosteum. This cell culture system responded to in vivo administered radiation by a significant decrease in cell numbers. This in vitro model will be subsequently used to study the cellular mechanisms of radiation and radioprotection in craniofacial osteoblast-like cells. PMID- 17912080 TI - Intracellular and extracellular tumor pH measurement in a series of patients with oral cancer. AB - Reversed pH gradient is an intrinsic feature of tumor phenotype resulting from an upregulation of glycolysis. This is crucial for tumor cell proliferation, invasion, metastasis, drug resistance, and apoptosis. Comprehension of mechanisms of pH regulation in tumors is of paramount importance for therapeutic implications. This is a preliminary report of a larger prospective study dedicated to the measurements of neutral or slightly alkaline pH/extracellular pH (pHi/pHe) in human patients affected by tumors of the head and neck. During surgery, four specimens were obtained from six patients with cancer: two from the tumor site and two from contralateral areas or sane areas near the tumor. pHe and pHi were measured and compared within normal and neoplastic tissues. Our data indicate that human spontaneous tumors show similar reversed gradients as observed in previous analysis on animal tumor models and cell lines. PMID- 17912081 TI - Parotid gland mucoepidermoid carcinoma associated with myasthenia gravis. AB - Myasthenia gravis (MG) is an autoimmune neuromuscular disease. Although most MG cases do not have accompanying malignant disease, patients with thymoma may have an increased risk for secondary malignancy. The reason MG patients are prone to developing cancer is not very well known. Salivary gland tumors account for approximately 3% of head and neck tumors, and approximately 80% of these occur in the parotid gland. Mucoepidermoid carcinoma is the most common malignant tumor of the parotid gland. MG may be associated with squamous cell carcinoma of the head and neck. We present a patient with MG who developed parotid gland mucoepidermoid carcinoma. MG patients with parotid gland cancer have not been previously reported. We also discuss the possible mechanisms of tumor development in an MG patient. PMID- 17912082 TI - Primary oral malignant melanoma. AB - Malignant melanoma of the oral cavity is a rare neoplasm. It is well known for his poor prognosis and the need for an evidence-based treatment. Therapy is commonly based on surgical excision of the primary tumour, supplemented by adjuvant therapy. In this paper two new cases of primary oral melanoma are reported with a review of the literature. PMID- 17912083 TI - Kimura disease: diagnostic imaging findings and surgical treatment. AB - We have encountered 11 cases of Kimura disease, comprising 10 males and 1 female. The ages at presentation ranged from 16 to 48 years, with a mean of 31.5 years. The sites of the subcutaneous masses were bilateral posterior auricular regions in two cases, frontal region in two cases, temporal region in two cases, head region in one case, parotid region in two cases, parotid and temporal regions in one case, and left earlobe in one case. The interval from onset to surgery ranged from 1 to 10 years, with a mean of 4.7 years. For diagnostic imaging, a combination of magnetic resonance imaging (MRI) and ultrasonography had a high diagnostic value. MRI depicted abnormalities at sites in contact with bone, such as posterior auricular regions, and sites with abundant soft tissue, such as parotid and cheek regions. Diffuse atrophy of subcutaneous fat was observed at the sites of the masses. On ultrasonography, the interior of lymph nodes was homogeneous and hyperechoic, whereas the periphery was hyperechoic, and blood vessels entering lymph nodes were clearly depicted. Surgery was performed in all cases. Postoperative adjuvant radiotherapy was conducted in one patient and radiotherapy and steroid therapy in one other patient. There were two relapses, and both were excised by repeated surgery. Surgical excision of the subcutaneous mass in Kimura disease has the advantages that the treatment period is short and precise histopathologic diagnosis can be obtained. PMID- 17912084 TI - Ozone therapy in extractive surgery on patients treated with bisphosphonates. AB - It is certain that oral extractive surgery is a remarkable trigger to avascular osteonecrosis of the jaw in patients treated with pyrophosphate analogous. This acquisition limits the use of endo-oral surgery in those patients, even when they have already developed the lesions. In this study, we present the results obtained in a group of 15 patients deriving from a 33-patient cluster with osteonecrosis of the jaw in treatment at our department with a new protocol based on ozone therapy. The object of this article is to demonstrate how dental extraction becomes possible in a patient with avascular bisphosphonate-related jaw osteonecrosis or in those who simply received pyrophosphate analogous when proper treatment with ozone therapy has been done. PMID- 17912085 TI - Ozone therapy in the treatment of avascular bisphosphonate-related jaw osteonecrosis. AB - Avascular osteonecrosis of the jaw (ONJ) may occur as a consequence of several conditions, even including chemotherapy treatment in patients affected by tumors or osteoporosis. We report our clinical experience in treating bisphosphonate induced ONJ with a therapeutic methodology that includes ozone therapy as a new and original approach for the clinical management of maxillary necrotic lesions. Of 58 patients with ONJ observed at our department, 33 gave their informed consent to be part of the research and were treated according to a therapeutic approach, which included noninvasive surgery associated with pre- and postsurgical cycles of ozone therapy consisting of eight sessions lasting 3 minutes each besides antibiotic and antifungal therapies. Outcomes showed how ozone therapy increases the benefits of surgical and pharmacologic treatments, increasing the complete healing of the lesions with the disappearance of symptoms and brings cases of lesion progression down to zero. In conclusion, ozone therapy is a reliable presidium in treatment of ONJ; its benefits are remarkable and improve significantly the outcomes of the surgical approach. PMID- 17912086 TI - Unusual presentation of human giant orf (ecthyma contagiosum). AB - Giant orf is a zoonotic infection that is endemic in sheep and goats. It may be transmitted to humans by direct contact with infected animals or contaminated objects and is typically found on the hands. We report the case of a sheep farmer with facial orf that proliferated dramatically with the formation of satellite lesions after curettage. PMID- 17912087 TI - Clinical parameters in T1N0M0 lower lip squamous cell carcinoma. AB - This retrospective study was carried out to asses the clinical outcome of T1 (i.e., tumor 2 cm or less at greatest dimension) squamous cell carcinoma of the lower lip (SCCLL) and verify the impact of some clinical parameters on prognosis. Fifty-seven patients with histologically proven T1 SCCLL were analyzed. Fifty-two patients were never treated before admission, whereas five (8.8%) had a second radical resection of the primary tumor location; none had neck nodes (i.e., N0) or distant metastasis (i.e., M0). The global disease-specific survival rate at 32 months was 100%, irrespective of grading and type of surgery, and thus no differences were statistically detected. Therefore, we concluded that radical tumor resection is a viable procedure for T1 SCCLL, irrespectively of grading. In addition, a second surgery on the primary tumor location is possible and has effectiveness on survival. Finally, neck dissection is not necessary in cases of T1 SCCLL. PMID- 17912088 TI - A congenital true teratoma with cleft lip, palate, and columellar sinus. AB - Teratomas, the most common extragonadal germ cell tumor of childhood, involve at least two of the ectodermal, mesodermal, and endodermal layers. Of the teratomas seen in the first 2 months of life, 82% are sacrococcygeal. The head and neck region is the second most common location for teratomas in early infancy, accounting for five (14%) of those cases. We describe a female neonate with a teratoma of the nasopharyngeal area, bilateral cleft palate/lip, and columellar sinus pathologies. The mass, which was 8 x 5 x 7 cm and soft in consistency, blocked the airway and prevented oral feeding. On macroscopic examination of the excised mass, there was a notable typical cilia arrangement and lower eyelid appearance. The patient, who was diagnosed with a well-differentiated teratoma after the pathologic examination, did not have any complications in the postoperative period. PMID- 17912089 TI - Facial defects restored with extraoral implant-supported prostheses. AB - A facial defect results in cosmetic, functional, and psychologic problems and acts as a difficult and challenging procedure for the maxillofacial surgeon and the prosthodontist. Retention of a facial prosthesis is an important key in the patient's satisfaction and acceptance. The introduction of extraoral implant applications have been accepted as an advanced retention method. The purpose of this article is to report the results from our clinical experience with patients wearing facial prostheses anchored to extraoral ITI implants (Straumann, AG, Basel, Switzerland). PMID- 17912090 TI - Intraorbital arteriovenous malformation: issues in surgical management. AB - Intraorbital arteriovenous malformations are infrequent but, when present, typically require complex surgical management. Computed tomography and magnetic resonance imaging help characterize lesion structure, location, and relationship to intraorbital anatomy. Frequently complex, intervention should be prompted by significant proptosis, increased intraocular pressure, pain, bleeding, ulceration, or high-output cardiac complications. For more controlled surgical debridement, angiographic embolization may help occlude the nidus and feeding vessels temporarily. Utmost caution must be taken to avoid injury to the optic nerve and ophthalmic artery. We present a rare case of complex, intraorbital arteriovenous malformation, as well as discuss aspects of management and relevant literature. PMID- 17912091 TI - Ankyloglossum superius syndrome: diagnosis and surgical management. AB - Ankyloglossum superius (or ankyloglossia superior) is a rare congenital craniofacial condition characterized by tongue tip adherence to the hard palate and associated limb abnormalities. There have been few reports of this condition in the literature. We present a patient with ankyloglossum superius syndrome and discuss diagnosis and surgical management. PMID- 17912092 TI - Piezosurgery: a new method for osteotomies in rhinoplasty. AB - Two basic techniques for lateral osteotomy have been developed to date; the internal (endonasal) continuous technique and the external (percutaneous) perforating method. Numerous investigators have subjectively reported that the application of the two techniques results in less postoperative ecchymosis and edema compared to the use of other techniques, but an alternative and gentle method for performing lateral osteotomy or bony hump removal has not been proposed yet. The authors present a new soft technique to perform nasal osteotomy in rhinoplasty using piezoelectric ultrasonic vibrations, and emphasize the advantages of this method. PMID- 17912093 TI - Extended approach in head and neck tumors: modified Schobinger incision. AB - Surgical treatment of many tumors of the head and neck require extended approaches. Many cervicotomies have been described in the treatment of head and neck and cervical tumors; careful thought must be made by a surgeon when surgical treatment is planned for an extended tumor.Our purpose is to show the versatility of the modified Schobinger incision. Between 2003 and 2005, 18 patients were operated on at our plastic surgery department; all presented extended cervical tumors of various origins.In all 18 patients, a Schobinger incision as modified by Yoel and Linares was used. The principal modification described by these authors consisted in the extension of the vertical line of incision below the subclavicular area to the nipple-areola complex. Partial necrosis occurred in two (11%) patients; one (1.8%) patient developed a pharyngocervical fistula; in all cases, healing occurred spontaneously. The benefits of this incision are as follows: 1) an unobstructed vision of anatomic landmarks and the more distal surgical elements; 2) increased blood supply for the flap by means of recruitment of new arterial vessels from the first, second, third, and fourth internal mammary perforators; 4) should flap necrosis occur, it will not compromise the coverage of the great vessels of the neck; 5) the flap's excessive mobility permits it to be used in facial areas, thus facilitating their reconstruction. Other advantages of this incision are that it permits access to different areas of the surgical field, that it allows protection of vital anatomic structures in both the pre- and postoperative periods, and that it results in an inconspicuous and aesthetically acceptable final scar. PMID- 17912094 TI - Versatility of frontal island flaps in the reconstruction of periorbital soft tissue defects. AB - The reconstruction of large soft tissue defects in the orbital and maxillomalar region is a challenging task that necessitates the consideration of both functional and aesthetic outcomes. We used the frontal island skin flap in patients with full-thickness soft tissue defects of the periorbital and malar region. In the reconstruction of full-thickness defects of this particular region, the alternatives to this flap are other regional flaps or distant free flaps. Not every pedicled regional flap can be transferred to every defect and most of the time the application of distant free flaps increases the morbidity of the procedure. The surgeon must be capable of being able to select the most useful and comprehensive flap within a range of many alternatives. We present our experience in 10 patients with complex soft tissue defects in the maxillomalar and periorbital regions whose defects were reconstructed with frontal artery island skin flaps. PMID- 17912095 TI - Supraorbital artery island flap for periorbital defects. AB - The forehead skin has the same color and texture as the periorbital region as well as the other parts of the face. The forehead is a local flap donor area for the reconstruction of full-thickness periorbital defects. This report presents eight cases in which full-thickness defects resulting from tumor resection have been repaired with supraorbital artery island flaps. Of eight patients, one was female and the rest were male with a mean age of 72.8 years (range, 64-88 years). Defects were located in the medial canthal region, lateral canthal region, glabella, and lateral part of the orbita. The flaps ranged from 2 x 3 cm to 6 x 7 cm in size. The patients were followed for 7 to 18 months. No complications occurred, except for decreased sensation on the forehead, and trapdoor deformity was seen in one case. The outcome was functionally and aesthetically satisfactory in all cases and all patients were happy with the outcome. The supraorbital artery island flap is a good alternative for the repair of defects around the orbita in that the color and texture of this flap match up with the orbital region and that it is pliable, simple, safe, and sensorial and requires only a single-session procedure. PMID- 17912096 TI - Z plasty closure of lower lip defects after tumor excision. AB - Reconstruction of the defects of the lower lip should provide a sensate, functional, and aesthetic lip structure. Defects of the lower lip, up to 30% of the total width, can be closed primarily, which gives a better result than any known flap operation unless the contracture of the linear scar tissue distorts the anatomic landmarks. Taking this possibility into consideration and to prevent scar contracture, we have performed Z plasty to the skin component while closing the resultant central defects of the lower lip after tumor excision. This modification improved our cosmetic results. PMID- 17912097 TI - Arterial vascularization of the extraocular muscles on its importance for orbital approaches. AB - The anatomic description of the arterial supply of the muscles of the eyeball was studied because medial and lateral approaches to the orbit to treat various disorders might severely damage the main trunk of the ophthalmic artery. The different arterial pedicles for each muscle, as well as their origins and points of penetration into the muscle, were studied in 19 male human orbits. In all cases, the muscular branches originated from the ophthalmic artery, and generally from the inferior face of this artery, just after crossing the optic nerve. The inferomedial muscular trunk arose from the distal of the ophthalmic artery (63.16%). The inferolateral muscular trunk, the superior oblique, levator palpebrae superioris, superior rectus, lateral rectus, and medial rectus arose from the lacrimal artery (43.36%), the bend of the ophthalmic artery (36.84%), the supraorbital artery (36.84%), the distal end of the ophthalmic artery (52.6%), the lacrimal artery (89.47%), and the inferomedial muscular trunk (84.51%), respectively. Complications resulting from damaging the artery can cause inadequacy in eye position, mobility, superior oblique function, or binocular vision, which are expected to be corrected postoperatively. PMID- 17912098 TI - Reconstruction of a severe maxillofacial deformity after tumorectomy and irradiation using distraction osteogenesis and LeFort I osteotomy before vascularized bone graft. AB - We present the successful reconstruction of a large mandibular defect with a severe maxillofacial deformity after malignant tumor resection and irradiation. The patient was a 16-year-old boy with a defect in the left mandible, which extended from the mandibular body to the condylar process and hypoplasia of the maxillozygomatic complex on the left side as a result of ablation and radiotherapy of a grown rhabdomyosarcoma in the left infratemporal fossa at the age of 10. We planned a two-stage reconstruction because of his wide mandibular defect and hypoplasia. LeFort I type osteotomy to correct the maxillary declination was combined with mandibular lengthening to decrease the width of the defect in the first stage. New bone formation was confirmed at the distraction site 4 months after surgery, and the second stage was performed. A free latissimus dorsi myocutaneous flap with a vascularized scapula and rib was transferred to reconstruct the ramus of the mandible, zygomatic arch, and soft tissues. This procedure resulted in satisfactory results. In conclusion, the combination of distraction osteogenesis and microsurgical bone transplantation facilitated the straightforward reconstruction of a three-dimensional deformity with huge bony defects. We think that this combined surgical procedure will become a favorable option in the treatment of severe maxillomandibular deformities with bone defects. PMID- 17912099 TI - A custom-made sealing screw cap for a closed, hollow obturator. AB - This paper describes a closed, hollow obturator with a nondetachable screw cap to seal the access hole that also allows cleaning of the inside of the bulb. A small screw cap was fabricated from a cobalt-chromium alloy. This screw was screwed to the access hole prepared at the top of the hollow obturator and when removed allows the inside of the obturator to be cleaned through the access hole. At the end of the screw, a preformed cobalt-chromium wire was attached that works as the safety retention mechanism of the screw. This ensures the screw in not aspirated or swallowed if it becomes unscrewed and falls from the prosthesis during wear in the patient's mouth. This device enables the patient to clean the inside of the closed obturator at home. PMID- 17912100 TI - Minimally invasive video-assisted submandibular sialoadenectomy: a preliminary report. AB - Open-field sialoadenectomy is the first-choice treatment for submandibular disease. Conventional surgery is based on the transcervical approach. The present study aims to demonstrate the feasibility of a minimally invasive video-assisted sialoadenectomy and to describe a new technique to perform it. A 52-year-old man affected by chronic sialoadenitis underwent minimally invasive video-assisted sialoadenectomy under general anesthesia. The ultrasound scanning of the submandibular gland showed a 6.5-cm hypertrophic gland with dilatated intraglandular ducts and a 2-cm long intraglandular sialolith. After surgery, no static or dynamic nerve deficiencies were detected, but slight temporary deficiency of the marginalis mandibulae nerve recovered in 15 days. Cosmetic results satisfied the patient. Minimally invasive video-assisted submandibular sialoadenectomy can be a feasible and safe procedure, which could be a valid choice to conventional surgery if performed on selected cases. More experience must be collected to analyze the cost-effectiveness. PMID- 17912101 TI - W(h)ither orbital pseudotumor? AB - Orbital pseudotumor is a nonspecific, idiopathic benign inflammatory process characterized by a polymorphous lymphoid infiltrate with varying degrees of fibrosis. We retrospectively reviewed the clinical presentation, management, and progress of six consecutive patients referred to our service with an initial diagnosis of orbital pseudotumor to underscore the challenge and pitfalls in managing this group of patients. Three male and three female patients, aged 27 to 74 years, presented with a variety of ophthalmologic problems, including orbital swelling, chemosis, proptosis, blepharoptosis, restricted eye motion, diplopia, and visual loss. The initial diagnosis of orbital pseudotumor was based on clinical findings, results of routine laboratory screening tests, computed tomographic and/or magnetic resonance imaging scans, and the response to corticosteroid treatment in three patients. In these three patients, the final diagnosis of orbital pseudotumor was confidently made only in one patient who remained in remission after corticosteroid therapy. The remaining two patients had Miller-Fisher syndrome and thyroid ophthalmopathy. Three other patients initially diagnosed with orbital pseudotumor underwent biopsy through an orbitotomy with comprehensive histopathologic evaluation. Two of these patients were subsequently diagnosed with non-Hodgkin lymphoma. Orbital pseudotumor belongs to a spectrum of lymphocytic infiltrative orbital conditions. It is a diagnosis of exclusion. The initial diagnosis must be regarded as provisional, and failure of complete resolution with corticosteroid therapy should heighten the index of suspicion and a biopsy should be considered. However, diagnosis may be difficult even with comprehensive histopathologic studies. PMID- 17912102 TI - Maxillary sinus lipoma: an unanticipated diagnosis. AB - Subcutaneous tissue gives rise to numerous lesions such as lipoma, the most common benign soft tissue tumors. Lipoma is seen only extremely rarely in osseous units. In addition, craniofacial involvement of intraosseous lipoma may be misdiagnosed as a fibroosseous tumour such as fibrous dysplasia. Here, we present a case of an intraosseous lipoma obviously destroying and invading the maxillary bone. PMID- 17912103 TI - Submandibular area metastasis from prostate small cell carcinoma with neuroendocrine differentiation. AB - Neuroendocrine differentiation in prostatic carcinomas generally confers a more aggressive clinical behavior and less favorable prognosis than usual prostatic carcinomas. In this article, we report a case of a 65-year-old man with prostatic carcinoma who had a metastasis of the submandibular area. His serum prostate specific antigen level was reduced to below the normal range and carcinoembryonic antigen was increased. Pathologic specimens demonstrated a small cell carcinoma with neuroendocrine differentiation by immunohistochemical studies. PMID- 17912104 TI - Controlled study of lactoperoxidase gel on oral flora and saliva in irradiated patients with oral cancer. AB - The aim of this study was to determine if radiotherapy induces hyposalivation altering oral microbial flora. The purpose of this placebo-controlled, single blind study was to determine beneficial effects of a saliva substitute and an oral hygiene product on irradiated patients with oropharyngeal cancer. Eighteen patients were assigned to the test group (Biotene Oral Balance gel [Laclede Incorporated Healthcare Products, Gardena, CA] and toothpaste used daily), and another 18 were put on a conventional daily regimen (carboxymethylcellulose gel and Oral-B toothpaste [Laclede Pharmaceuticals, Gardena, CA]). Cultures for identifying and quantitating microorganisms, whole unstimulated saliva, and visual analog measurements for comfort were obtained before mucositis occurred and after treatment. Daily use of Biotene products enhanced control of microbial flora, improved salivary flow, and increased oral comfort as compared with control subjects. Four weeks after mucositis, some aerobic isolates disappeared in the test group; periodontal-associated bacteria were markedly decreased in the test group; and candidal species were significantly lowered in the test group. Although baseline saliva was lower in the test group (P = 0.001), after 4 weeks, no difference between groups existed; comfort was greater in the test group (P = 0.007). Use of enzyme-engineered Biotene products that assist in control of the oral microbial flora as well as supporting oral comfort through lubrication appear to be useful aids for irradiated patients with oropharyngeal cancer. PMID- 17912105 TI - Combined surgical excision and radiation therapy for keloid treatment. AB - Various methods have been attempted for the treatment and management of keloids; however, there is little satisfactory clinical evidence in long-term follow ups. Also, there is a preference for occurrence and recurrence in anatomic location. Usually anatomic locations with higher regional tension and more sebaceous glands are inclined toward pathogenesis. Thirty-eight keloids treated with combined surgical excision and postoperative irradiation, using electron beams with only a 10-mm opening by lead shielding, were investigated at a mean follow up of 4.4 +/- 2.5 years (range, 1-9 years) at a single institute. Ten locations such as the ear (n = 6), neck (n = 3), and upper lip (n = 1) were among the craniofacial locations. The hardness of the keloids and posttreatment scars was clinically and objectively tested with the Vancouver scar scale and a durometer, which is often used for the industrial measurement of thread balls and rubber. At a mean of 4.4 +/- 2.5 years of follow up, the clinical characteristics of the scars were significantly better posttreatment as 2.6 +/- 0.5 versus 1.0 +/- 0.6, 3.7 +/- 0.7 versus 1.7 +/- 0.7, 2.9 +/- 0.4 versus 1.3 +/- 0.5, and 2.7 +/- 0.5 versus 1.3 +/ 0.5 (keloid scars versus posttreatment scars: pigmentation, pliability, height and vascularity, respectively, P < 0.01). The durometer readings were significantly lower posttreatment, 15.2 +/- 3.9 versus 7.7 +/- 2.9 (keloid scars versus posttreatment scars, P < 0.01). The recurrence rate was 21.2% overall with none in craniofacial locations. Therefore, the combined treatment of surgical excision and postoperative electron beam irradiation is effective for scar quality and reducing the recurrence rate in long-term follow up. PMID- 17912106 TI - Peripheral osteoma of the maxillary alveolar process. AB - Osteoma is a benign, slow-growing tumor characterized by proliferation of compact or cancellous bone. Solitary osteomas are classified as peripheral, central, or extraskeletal. Peripheral osteomas of the oral cavity are unusual and the maxilla is rarely affected. They manifest as asymptomatic, fixed tumors of bony-hard consistency that may be sessile or pedunculated. Radiographically, a well circumscribed round or oval radiopaque mass is seen that is microscopically composed of cancellous or trabecular bone. A case of a compact, peripheral osteoma arising from the buccal plate of the alveolar ridge of the maxilla in a 64-year-old patient is presented. According to our literature review, this is the seventh case reported in the maxillary ridge. PMID- 17912107 TI - The importance of the head and neck region in regression of advanced MCC: a clinical report. AB - This paper presents the case of a 76-year-old woman who experienced a total regression of a Merkel cell carcinoma (MCC). The primary site of the tumor was on her right eyebrow. After this lesion was excised, the patient presented a massive locoregional metastasis on the right parotid gland and the laterocervical lymph nodes. No distant metastases were detected. An incisional biopsy into the right parotid gland confirmed the diagnosis of MCC metastasis. No surgical treatment was prescribed because of the advanced stage of the disease. Spontaneous total regression on the parotid and the neck mass was observed within 3 months. This is the 15th case of spontaneous regression in total and the 14th case with a site of origin in the head and neck region. PMID- 17912108 TI - Osteolipoma of the nasopharynx. AB - Osteolipomas of the head and neck are very rare, and there is no reported case of osteolipoma of the nasopharynx in the literature. Here, we present a case of osteolipoma of the nasopharynx in a 21-year-old male who was referred for a nasopharyngeal mass causing a defect in the left half of the soft palate. The well-demarcated, firm tumor was excised through transnasal endoscopic and transpalatal approaches, and the defect of the soft palate was repaired by a superiorly based pharyngeal flap. Osteolipomas should be kept in mind in the differential diagnosis of nasopharyngeal mass as an extremely rare benign lesion. PMID- 17912109 TI - Correction of saddle nose deformity in ectodermal dysplasia. AB - Ectodermal dysplasia (ED) is a group of disorders affecting ectodermal-derived tissues, mainly the skin and its appendages, including teeth, nails, hair, and sweat glands. Along with unusual and typical craniofacial findings, these patients present with a unique nasal deformity. This consists of wide nasal base, saddle nose deformity, lack of alar grooves, and wide nostrils. We present a case of ED in which the nasal deformity was treated with bilateral nasal osteotomies, costochondral cartilage graft to the dorsum, and reforming of alar grooves by suturing. PMID- 17912110 TI - Warthin's tumor of the hard palate. AB - Warthin's Tumor (WT) or papillary cystadenoma lymphomatosum is typically located in parotid gland. Extraparotid localization of WTs (EPWT) was rarely described in ectopic salivary tissue in the latero-cervical and para-parotid lymph nodes. Localization in the hard palate is exceptional. This paper describes pre-surgical diagnostic work up and surgical management of a solitary WT localized in the hard palate as found in a 27-year-old Caucasian woman. With consideration of uncertain pre-surgical diagnosis obtained with FNAC, intraoperative histologic examination of frozen sections was planned. Conservative surgical approach was performed. After WT was diagnosed with histology, a local bone curettage was made. Clinical and radiographic follow-up at 12th month was negative for relapse. PMID- 17912111 TI - Hyoid chondroma presenting as an external neck mass. AB - Chondromas of the larynx are rare neoplasms; only a few examples of cartilaginous tumors affecting hyoid bone have been reported. A 33-year-old woman presented with a neck mass on the left carotid triangle. The patient's computed tomography and magnetic resonance imaging revealed a heterogenous mass, which seemed to originate from the left greater cornu of hyoid. The mass was excised using a transcervical approach with left greater cornu of hyoid bone. The pathologic diagnosis was chondroma of hyoid. Expert radiologic and pathologic review is mandatory in cartilaginous neoplasms of the larynx. Cartilaginous neoplasms of the hyoid should be included in the differential diagnosis of neck masses at the carotid triangle. PMID- 17912112 TI - Recurrent granular cell tumor: how to treat. AB - Granular cell tumor is an uncommon lesion usually located in the head and neck region (50%) with a female sex predominance. It is believed to be of primitive neuroectodermal origin. Typical clinical presentation is a small, uninflamed, slowly growing, yellowish mass approximately 2 cm in diameter. Granular cell tumor has a peculiar clinical behavior ranging from clearly benign, locally aggressive, or manifestly malignant. Treatment is surgical excision, but recurrences are possible. In this article, we present a case of recurrent benign granular cell tumor with the discussion of the treatment modalities. The patient was a 16-year-old girl with a 3-year history of a painless mass on the right side of her neck. The mass was excised two times at different centers and diagnosed as granular cell tumor after the pathologic examination. The time interval between the operations and recurrences was approximately 1 year. On physical examination, a yellowish, firm, nodular mass measuring approximately 2 x 1 cm was noted. Cervical lymph nodes were evaluated with ultrasonography preoperatively to exclude metastasis and no pathologic lymph nodes were noted. The lesion was excised under local anesthesia with a 1-cm safe margin and the resulting defect was closed primarily after undermining of the wound edges. Postoperatively, the wound began to heal with a hypertrophic scar and immediate precautions were taken. Topical steroid treatment and silicone blocks were applied. Now the patient is in the sixth postoperative month and followed up every month for signs of recurrence. PMID- 17912113 TI - Primary mucosa-associated lymphoid tissue lymphoma of hypopharynx. AB - Mucosa-associated lymphoid tissue lymphomas are low-grade B-cell lymphomas that arise from a number of extranodal sites, including both nonmucosal and mucosal organs such as the hypopharynx. We reported a patient with a primary hypopharynx mucosa-associated lymphoid tissue lymphoma presenting with a swallowing dysfunction and severe throat pain. The clinical, radiologic, and histopathologic findings are presented. The patient was followed up for 5 years and treated with nonspecific antibiotics, chemotherapy, and radiation therapy. Because of prevertebral fascia invasion at the initial presentation, surgical treatment was not preferred. The last biopsies of the hypopharynx revealed no evidence of lymphoid infiltrate. Mucosa-associated lymphoid tissue lymphoma involving the hypopharynx is rare and there is no consensus on its treatment. The treatment protocol is presented and the relevant literature is reviewed. PMID- 17912114 TI - Concurrent large spindle cell and ordinary lipomas. AB - The number of reported coexistences of ordinary lipoma and spindle cell lipoma is very limited in the literature. We present a patient with neighboring large spindle cell lipoma and ordinary lipoma in the posterior neck. Clinical, radiologic, and histologic findings of the case are presented with distinguished figures. Complete surgical removal confirmed the diagnoses. These lipomatous tumors may develop coincidentally and magnetic resonance imaging is overtly adequate in identifying their nature. PMID- 17912116 TI - Adult orthognathic surgery. PMID- 17912117 TI - Comparison of treatment outcome and stability between distraction osteogenesis and LeFort I osteotomy in cleft patients with maxillary hypoplasia. AB - The purpose of this study was to compare treatment outcome and relapse between maxillary advancement surgery with LeFort I osteotomy and maxillary distraction osteogenesis in patients with cleft lip and palate with maxillary hypoplasia. The sample consisted of a maxillary advancement surgery with LeFort I osteotomy group (group 1, N= 14, mean age, 21.7 years) and a maxillary distraction osteogenesis group (group 2, N = 11, mean age, 16.3 years). Lateral cephalograms were taken and traced at presurgery (T0), postsurgery (T1), and postretention (T2). Nine hard and four soft tissue cephalometric variables were measured. Differences in measurements at each stage, treatment outcome (T1-T0), and relapse (T2-T1) were compared between groups with independent t test. Because the amount of surgical movement could affect the amount of relapse, a difference in relapse between two groups was compared by analysis of covariance with the amount of surgical movement as a covariant. Although the amounts of forward movements of A point (P < 0.01), upper incisor (P < 0.001), and upper lip (P < 0.001) during T1-T0 were greater in group 2, there were no significant differences in the amounts of relapse (T2-T1) between the two groups. During T1-T0, counterclockwise rotation of the palatal plane was observed in group 2 as a result of downward movement of posterior nasal spine (PNS) at T1, whereas group 1 had clockwise rotation of palatal plane at T1 because of downward movement of anterior nasal spine (ANS). The amounts of relapse (T2-T1) in vertical movements of PNS and upper incisor were significantly different between the two groups (P < 0.05). The amount of required maxillary advancement, vector control of palatal plane, and vertical position of upper incisor would be important factors when planning a surgical treatment in patients with cleft lip and palate with midface hypoplasia. PMID- 17912118 TI - Segment stability in bimaxillary orthognathic surgery after resorbable Poly(L lactide-co-glycolide) versus titanium osteosyntheses. AB - This study compared segment stability after bimaxillary orthognathic surgery, comparing poly(L-lactide-co-glycolide) with titanium osteofixation at 12 months follow up. Fifteen patients were osteofixated with poly(L-lactide-co-glycolide) copolymer, 30 with 2.0-mm titanium miniplates. Preoperative, postoperative, and 1 year follow-up lateral cephalograms were analyzed. Maxillary average advancement in resorbable plate osteosyntheses (+/- standard deviation) was (case numbers/titanium controls) 2.5 (+/- 1.0) mm; n = 7/5.4 (+/- 3.5)mm; n = 21, setback 2.2 (+/- 2.4) mm; n = 7/1.9 (+/- 1.8) mm; n = 8, elongation 6.5 (+/- 3.4) mm; n = 9/3.7 (+/- 5.2) mm; n = 14, intrusion 1.0 (+/- 0.7) mm; n = 5/3.3 (+/- 2.7)mm; n = 13, mandibular average advancement was 5.5 (+/- 3.7) mm; n = 4/6.3 (+/- 8.8) mm; n = 18, setback 11.2 (+/- 7.7) mm; n = 7/7.2 (+/- 3.2) mm; n = 12, mandibular angle enlargement 7.9 (+/- 2.4) degrees ; n = 9/7.9 (+/- 6.6) degrees ; n = 21, reduction 6.9 (+/- 2.6) degrees ; n = 4/6.3 (+/- 6.6) degrees ; n = 9. Changes in landmark position within the study and control groups differed significantly in paired t testing (P =.01); operative movements were comparable in between study and control groups (P = 0.5, two-sided t test), only maxillary advancement was significantly smaller (P = 0.04) within study cases. Absolute instability at advanced A-point was (study group/controls) 1.2 (+/- 0.8)/2.4 (+/- 2) mm; setback 1.8 (+/- 1.9) mm/2.5 (+/- 1.7) mm; elongation at anterior nasal spine (ANS) 2.0 (+/- 1.4) mm/3.1 (+/- 3.6) mm, intrusion 1.1 (+/- 1.1) mm/2.2 (+/ 1.5) mm; advancement instability at B-point was 2.6 (+/- 2.7) mm/5.1 (+/- 8.2) mm, setback 2.7 (+/- 2.6) mm/1.7 (+/- 2) mm; mandibular angle enlargement instability 2.4 (+/- 2.7) degrees /8.2 (+/- 9.6) degrees , angle narrowing 7.0 (+/- 5.4) degrees /4.2 (+/- 5.9) degrees . Absolute postoperative instability was not significantly different in between study and control groups (P = 0.3). Tested resorbable poly(L-lactide-co-glycolide) osteofixation proved to be as reliable in segment fixation as titanium; however, study and control groups were not matched; the study group was small and therefore the results (especially advancement) have to be interpreted as preliminary until larger prospective cohorts become evaluated. PMID- 17912119 TI - A novel model surgery technique for LeFort III advancement. AB - Current techniques for model surgery and occlusal splint fabrication lack the ability to mark, measure and plan the position of the orbital rim for LeFort III and Monobloc osteotomies. This report describes a model surgery technique for planning the three dimensional repositioning of the orbital rims. Dual orbital pointers were used to mark the infraorbital rim during the facebow transfer. These pointer positions were transferred onto the surgical models in order to follow splint-determined movements. Case reports are presented to illustrate how the model surgery technique was used to differentiate the repositioning of the orbital rim from the occlusal correction in single segment and combined LeFort III/LeFort I osteotomies. PMID- 17912120 TI - Quantitative study of new bone formation in distraction osteogenesis of craniofacial bones by bone scintigraphy. AB - Although distraction osteogenesis is widely accepted as a technique to augment the craniofacial skeleton, timing to start distraction after an osteotomy or to remove distractors is basically based on studies on long bones. Because bone scintigraphy is well known to be the gold standard for quantitative measurement of bone formation, we conducted this pilot study to evaluate its feasibility as a tool for assessing new bone formation by distraction osteogenesis. Five patients with midface hypoplasia and four with mandibular hypoplasia were studied. Each patient had five bone scans: before surgery, 3 and 30 days after stopping distraction, and 3 days before and 3 months after distractor removal. Radiotracer uptake values at distraction sites were measured at 1 and 3 hours. Each uptake value was compared with preoperative study as uptake ratio. A typical pattern of radiotracer uptake ratio was observed in all cases with successful distraction. Uptake rose to the maximum during the consolidation period and remained at or above the preoperative level until the study end point. In one patient who had mandibular distraction and nonunion of the right ramus, there was no uptake peak during early consolidation as seen in the successfully distracted body and in the other cases. Bone scintigraphy was found to be a useful investigation in craniofacial distraction. It showed the dynamic of new bone formation by demonstrating the osteoblastic activity, which is important objective information for determining distraction rate and consolidation duration in each case. It may also be a tool that can predict the outcome of distraction osteogenesis. PMID- 17912121 TI - Reflections on complications to bioresorbable osteofixation devices. PMID- 17912124 TI - Clinical outcomes of BRYAN cervical disc arthroplasty: a prospective, randomized, controlled, multicenter trial with 24-month follow-up. AB - STUDY DESIGN/SETTING: Prospective, randomized, 3-center, clinical trial. OBJECTIVE: To prospectively compare the outcomes of cervical arthroplasty with the BRYAN Cervical Disc Prosthesis (Medtronic Sofamor Danek, Inc, Memphis, TN) to anterior cervical discectomy and fusion (ACDF). SUMMARY OF BACKGROUND DATA: Surgical treatment of cervical disc pathology commonly involves techniques that employ discectomy and fusion (ACDF). This "gold-standard" technique has demonstrated good clinical and radiographic outcomes. Common adverse effects of this procedure are associated with the adjacent level degeneration and bone-graft harvest. Several investigators have independently reported successful short-term outcomes with the BRYAN Cervical Disc Prosthesis. In addition, a significant body of knowledge has been collected regarding the wear patterns and adjacent level effects of this device in human and animal models. METHODS: As part of an FDA IDE trial, 3 centers collected prospective outcomes data on 115 patients randomized in a 1:1 ratio to ACDF (Control group) or arthroplasty with the BRYAN Cervical Disc Prosthesis (Investigational group). RESULTS: Demographic and surgical data were generally similar in the 2 populations. Outcomes data collected at routine postoperative intervals for 24 months demonstrated that the Investigational group had statistically significant (P<0.05) improvements as assessed by the Neck Disability Index, the Neck Pain Score, and SF-36 Physical component scores. The improvement in the Mental Component Subscore values for the BRYAN and control groups was equivalent at 24 months (P=0.055). Arm pain relief was similar in both groups (P=0.152). During the course of the 2-year follow-up, 4 patients in the Control group required surgical intervention and 3 patients in the Investigational group required ACDF for adjacent level disease. CONCLUSIONS: At 24 months, cervical arthroplasty with the BRYAN Cervical Disc Prosthesis compares favorably with ACDF as defined by standard outcomes scores. PMID- 17912125 TI - Open-door laminoplasty with suture anchor fixation for cervical myelopathy in ossification of the posterior longitudinal ligament. AB - Expansive laminoplasty was developed to achieve posterior spinal cord decompression while preserving cervical spine stability. In the classic Hirabayashi procedure, the lamina door is tethered open by sutures between the spinous process and facet capsule or paravertebral muscle. The authors present a modified technique, which enhances secure fixation and prevents restenosis owing to hinge closure. Twenty-seven patients (7 females, 20 males) with cervical myelopathy secondary to ossification of the posterior longitudinal ligament were enrolled. Each patient underwent unilateral open-door laminoplasty with suture anchor fixation. Tying and fixation of the sutures onto the holed lateral mass screws was used instead of the conventional method. Radiography, magnetic resonance imaging, and computed tomography scanning were used for imaging studies. The Nurick score was used to assess myelopathy severity, whereas the Japanese Orthopedic Association score was adopted to compare clinical outcome before and after surgery. Mean follow-up period was 38 months (range, 18 to 60). Ten patients had 5 levels of decompression (C3-7), and 17 patients had 4 (C3-6, 12 patients; C4-7, 5 patients). All patients experienced functional improvement of at least 1 Nurick score after surgery. The Japanese Orthopedic Association score increased significantly from 7.5+/-3.2 before surgery to 13.2+/-1.6 at final follow-up. Postoperative radiography and computed tomography scan demonstrated significantly increased sagittal diameter and canal expansion. No neurologic deterioration owing to hinge reclosure or major surgery-related complications were observed. In conclusion, unilateral open-door laminoplasty with suture anchor fixation effectively maintains expansion of the spinal canal and resists closure while preserving alignment and stability. This modified technique has a low complication rate and provides marked functional improvement in patients with cervical myelopathy owing to ossification of the posterior longitudinal ligament. PMID- 17912126 TI - C1-2 transarticular screw fixation in high-riding vertebral artery: suggestion of new trajectory. AB - A significant drawback of atlantoaxial transarticular screw fixation is a potential risk of vertebral artery injury, especially with a high-riding type. The authors propose a relatively safe trajectory in cases of high-riding vertebral artery by using the 3-dimensional computerized tomography image reconstruction programs. Twelve consecutive patients with a pathologic condition in atlantoaxial complex were prospectively analyzed. Five other patients, whose high-riding vertebral arteries were incidentally found during the 3-dimensional computerized tomography performance for other cervical pathologic conditions, were also included. The preoperative screw simulation images, convergence angle, and caudal tilting angle for each screw were obtained from each patient. Of 17 subjects, 7 had high-riding vertebral artery unilaterally and 1 had bilaterally. All 12 patients with pathologic atlantoaxial complex, including 2 unilateral and 1 bilateral high-riding vertebral artery, had atlantoaxial transarticular screw fixation. For these 3 patients, the entry point and the trajectory for screw were moved more superiorly and medially as in cases with C2 pedicle screwing. The mean convergence angle and caudal tilt angle obtained during screw simulation for patients with high-riding vertebral arteries was 17.6 and 38 degrees compared with 21 and 53.3 degrees for patients with normal course of vertebral artery. It was possible to insert transarticular screws safely in patients with high-riding vertebral artery guided by preoperative screw insertion simulation program. PMID- 17912127 TI - Biomechanical comparison of unicortical versus bicortical C1 lateral mass screw fixation. AB - STUDY DESIGN: Biomechanical study of pullout strength of unicortical versus bicortical C1 lateral mass screws using a cadaveric cervical spine model. OBJECTIVE: To compare pullout strength of unicortical versus bicortical C1 lateral mass screws. SUMMARY OF BACKGROUND DATA: The internal carotid artery and hypoglossal nerve lie over the anterior aspect of the lateral mass of the atlas and are at risk from bicortical C1 lateral mass screws. Unicortical screws would reduce the risk of injury to these neurovascular structures; however, no data are available on the relative strength of unicortical versus bicortical C1 lateral mass screws. METHODS: Fifteen cadaveric cervical spine specimens underwent axial pullout testing of C1 lateral mass screws. A unicortical C1 lateral mass screw was placed on 1 side with a contralateral bicortical screw. RESULTS: The mean pullout strengths of the unicortical screws and bicortical screws were 588 N (range, 212 to 1234 N) and 807 N (range, 163 to 1460 N), respectively (P=0.008). CONCLUSIONS: Bicortical C1 lateral mass screws were significantly stronger than unicortical screws; however, the mean pullout strength of both the unicortical and bicortical C1 screws were greater than previously reported values for subaxial lateral mass screws. On the basis of these data, the clinical necessity for using bicortical screw fixation in all patients must be questioned. If similar strength can be achieved using unicortical C1 lateral mass screw to that currently accepted in the subaxial spine, bicortical screws might not be justified for the C1 lateral mass. However, the ability to extrapolate C1-C2 data to subaxial spine data is uncertain because of the difference in normal physiologic loading at these levels. PMID- 17912128 TI - Single-stage posterolateral vertebrectomy for the management of metastatic disease of the thoracic and lumbar spine: a prospective study of an evolving surgical technique. AB - Appropriate surgical management of spinal metastases combines maximal neural decompression with simultaneous immediate spinal column stabilization in the context of a paliative operation undertaken to improve patients' quality of life. We have used a single-stage posterolateral vertebrectomy (SPLV) for disease of the lumbar spine, combined with bilateral costotransversectomies in the thoracic spine, for these challenging cases. In this prospective cohort study of 96 consecutive patients with metastatic disease of the spinal column for we describe our surgical technique in detail, we examine our learning curve in its use and we analyze the long-term surgical and "quality of life" results in 42 patients who underwent SPLV. The mean and maximum operative blood loss was significantly lower for the SPLV group when compared with combined approaches. All patients either remained neurologically stable or had improved with surgery. Both the mean and the range visual analog scale scores were significantly improved after the SPLV. The SPLV was the only surgical approach to demonstrate a statistically significant improvement in Eastern Cooperative Oncology Group scores at 3 months after the surgery. Seventy-five percent of patients were alive at 6 months and 50% of patients survived for more than 12 months after the surgery. Eleven patients had a major complication (26%) with 9 (21%) patients required early reoperation, 7 of them for wound failure. Our data demonstrates that the SPLV represents a technically achievable improvement in surgical approach to spinal metastases when key parameters are examined. On the basis of these results, we recommend that the SPLV should be considered in all cases where resection of thoracic or lumbar spinal metastatic disease and reconstruction is contemplated. PMID- 17912129 TI - Clinical outcome after segmental wire fixation and bone grafting for repair of the defects in multiple level lumbar spondylolysis. AB - The aim of this retrospective study was to assess clinical outcomes after segmental wire fixation and bone grafting for repair of pars defects in patients with multiple-level lumbar spondylolysis. Subjects were 7 patients (5 men and 2 women, mean age 26.7 y) with multiple-level lumbar spondylolysis treated by segmental wire fixation and bone grafting at one of our affiliated institutions between 1983 and 2004. Clinical outcomes were determined by comparing preoperative and postoperative Japanese Orthopaedic Association scores and Mancab criteria, and healing of pars defects was evaluated by radiographic and computed tomography study. The condition involved 2 levels in 5 cases and 3 levels in 2 cases. The mean postoperative follow-up period was 51.0 months. The mean Japanese Orthopaedic Association score improved significantly from 21.29 before surgery to 27.86 after surgery, and the recovery rate was 85.21%. An "excellent" result was achieved in 5 cases, a "good" result in 1 case and a "fair" result in 1 case according to the Macnab criteria. Postoperative radiographs revealed healing of all defects in 4 cases, healing of 3 out of 4 defects in 2 cases, and no healing of any defect in 1 case. Pseudoarthrosis was related to wire breakage, and patients who did not obtain complete healing were patients who did not fully comply with instructions to wear a lumbar corset or restrict activity postoperatively. Segmental wire fixation and bone grafting were shown to be effective for multiple-level lumbar spondylolysis. PMID- 17912131 TI - C5 palsy after laminectomy and posterior cervical fixation for ossification of posterior longitudinal ligament. AB - OBJECTIVE: To investigate the imaging findings correlated with C5 root palsies in the patients undergoing laminectomy and lateral mass screw fixation for ossification of posterior longitudinal ligament (OPLL), and clarify its pathogenic mechanism. METHODS: The series included 49 patients with OPLL. Characteristics of preoperative and postoperative x-ray, computed tomography, and magnetic resonance images were compared between the patients with and those without C5 root palsies. RESULTS: Postoperative C5 root palsies occurred in 9 patients 6 to 64 hours postoperatively. They tended to have increased cervical lordosis and severe OPLL. However, there was no significant positive correlation with an increase in T2-weighted hyperintense foci on magnetic resonance studies. CONCLUSIONS: The tethering effect on the root seemed to be the main pathogenic mechanism of C5 root palsies in this study. PMID- 17912130 TI - Percutaneous treatment of lumbar intervertebral disk hernias with radiopaque gelified ethanol: a preliminary study. AB - STUDY DESIGN: Prospective clinical trial. OBJECTIVE: Demonstrate the safety and efficacy of gelified ethanol in the percutaneous treatment of lumbar disk hernias. SUMMARY OF BACKGROUND DATA: After the commercial withdrawal of Chymopapain, the need for new substances to treat intervertebral disk hernias was evident. Good results were obtained with pure ethanol, but this substance was difficult to handle. We decided to use a similar substance mixed with ethylcellulose to increase its viscosity and enhanced with radiopaque material. METHODS: Two hundred seventy-six consecutive patients sent to be treated of a lumbar intervertebral disk hernia percutaneously were included in this preliminary study and treated with radiopaque gelified ethanol (RGE) and intra articular steroids. Three groups were set, group A for patients to be treated only with RGE and groups B and C for difficult cases presenting a narrow canal, foraminal hernia, or hiperalgic sleepless hernia, treated with RGE plus another intradiscal technique, automatized percutaneous diskectomy for group B and radiofrequency nucleoplasty for group C. RESULTS: Very good or good results were obtained in 202 (91.4%) of the 221 patients in group A. Of the 44 patients in group B, 37 patients (84%) presented very good or good results and in 9 (82%) of the 11 patients of group C, we obtained similar results. There was no allergic complication in any of our patients. Short-term follow-up with magnetic resonance showed little or no changes in the intervertebral disk but there was discordance with clinical signs. Long-term follow-up magnetic resonance showed a dramatic reduction in hernia volume. CONCLUSIONS: This preliminary study shows the efficacy and inocuity of this new substance that could take over the Chymopapain therapeutic field. PMID- 17912132 TI - Acute hemorrhagic cyst of the ligamentum flavum. AB - STUDY DESIGN: A case report and clinical discussion. OBJECTIVE: To describe a rare complication of a cyst of the ligamentum flavum, which bled spontaneously, provoking an acute lower limb monoparesis and lumbar sciatic pain. SUMMARY OF BACKGROUNDS DATA: Cysts of the ligamentum flavum have been rarely reported. Intraspinal degenerative cysts described in literature are usually juxta articular (synovial and ganglion) cysts and have a similar radiologic appearance. They are preferentially located in the lumbar spine, while the cervical localization is unusual. Hemorrhage into the cyst is an uncommon complication and an extremely rare cause of nerve root compression. METHODS: A 59-year-old woman presented with sudden severe radicular lumbar deficit and pain secondary to acute hemorrhage into a ligamentum flavum cyst. Magnetic resonance imaging showed at L3 L4 level a lobulated slightly hyperintense mass with a ventral area of marked hyperintensity in T1 images, hypointense on T2 images. Signal within the lesion was suggestive of intralesional hemorrhage. RESULTS: Complete resection of the lesion was performed, resulting in immediate recovery. The cyst was quite rounded, brownish, and contained rest of both partially fresh and coagulated hematoma. Histologic examination revealed myxoid degeneration of the ligamentum flavum with an hemorrage in the cystic cavity without a synovial layer. CONCLUSIONS: This report identifies a rare case of radicular lumbar deficit and pain secondary to acute hemorrhage into a ligamentum flavum cyst. The pathogenesis and clinicopathologic characteristics of this lesion are described. PMID- 17912133 TI - Facet joint pain in chronic spinal pain: an evaluation of prevalence and false positive rate of diagnostic blocks. AB - STUDY DESIGN: A retrospective review. OBJECTIVES: Evaluation of the prevalence of facet or zygapophysial joint pain in chronic spinal pain of cervical, thoracic, and lumbar origin by using controlled, comparative local anesthetic blocks and evaluation of false-positive rates of single blocks in the diagnosis of chronic spinal pain of facet joint origin. SUMMARY OF BACKGROUND DATA: Facet or zygapophysial joints are clinically important sources of chronic cervical, thoracic, and lumbar spine pain. The previous studies have demonstrated the value and validity of controlled, comparative local anesthetic blocks in the diagnosis of facet joint pain, with a prevalence of 15% to 67% variable in lumbar, thoracic, and cervical regions. False-positive rates of single diagnostic blocks also varied from 17% to 63%. METHODS: Five hundred consecutive patients receiving controlled, comparative local anesthetic blocks of medial branches for the diagnosis of facet or zygapophysial joint pain were included. Patients were investigated with diagnostic blocks using 0.5 mL of 1% lidocaine per nerve. Patients with lidocaine-positive results were further studied using 0.5 mL of 0.25% bupivacaine per nerve on a separate occasion. Medial branch blocks were performed with intermittent fluoroscopic visualization, at 2 levels to block a single joint. A positive response was considered as one with at least 80% pain relief from a block of at least 2 hours duration when lidocaine was used, and at least 3 hours or longer than the duration of relief with lidocaine when bupivacaine was used, and also the ability to perform prior painful movements. RESULTS: A total of 438 patients met inclusion criteria. The prevalence of facet joint pain was 39% in the cervical spine [95% confidence interval (CI), 32%-45%]; 34% (95% CI, 22%-47%) in the thoracic pain; and 27% (95% CI, 22%-33%) in the lumbar spine. The false-positive rate with a single block in the cervical region was 45%, in the thoracic region was 42%, and in the lumbar region 45%. CONCLUSIONS: This retrospective review once again confirmed the significant prevalence of facet joint pain in chronic spinal pain. PMID- 17912134 TI - Correlation of spinal canal dimensions to efficacy of epidural steroid injection in spinal stenosis. PMID- 17912135 TI - Outcomes of inpatient rehabilitation for older adults with debility. AB - OBJECTIVE: To determine the functional outcomes and discharge setting of older patients receiving inpatient rehabilitation for debility (ICD-9-CM, 799.3). DESIGN: Retrospective cohort study of 63,171 individuals >or= 65 yrs old with a primary (23%) or comorbid (77%) debility diagnosis from the Uniform Data System for Medical Rehabilitation (UDSMR) database for 2002-2003. RESULTS: Patients with a primary diagnosis of debility (PDD) had a lower mean rehabilitation efficiency score (functional change per day) as compared with the rest of the subjects (1.7 vs. 1.9, P<0.001), including those with a Centers for Medicare & Medicaid Services (CMS) 75% rule primary diagnosis (1.8, P<0.001). The PDD group was less likely to be discharged home (68% vs. 73%, P<0.001) and more likely to be discharged to a hospital (13% vs. 11%, P<0.001). CONCLUSIONS: From a clinical perspective, the functional recovery of older patients with debility is essentially the same, regardless of whether this is a primary or comorbid diagnosis. Their functional improvement is also comparable with that reported for other CMS 75% rule diagnoses, although the debility patients are less likely to be discharged home. More than 10% of these patients were discharged to acute hospital settings. Further research is warranted to identify the most appropriate rehabilitation setting for patients with debility. PMID- 17912137 TI - Outcome assessment in randomized controlled trials of stroke rehabilitation. AB - The lack of a unified approach to outcome assessment in stroke rehabilitation limits our ability to interpret evidence provided by randomized controlled trials (RCTs). The purpose of this review was to identify outcomes and assessment tools reported in RCTs of stroke rehabilitation interventions as a first step toward consistent assessment of outcomes. Given that the validity of research is linked to reliability and validity of measurement, the relationship between the use of previously developed outcome measures and the methodological quality of RCTs was explored. Electronic literature searches identified RCTs examining stroke rehabilitation therapies from 1968 to 2005. The Physiotherapy Evidence Database (PEDro) scale was used to assess methodological quality. Cited outcomes were recorded and assessment tools identified as previously published or study specific. Four hundred ninety-one RCTs cited the assessment of 1447 outcomes using 489 measurement tools. Two hundred fifty-four of these were previously published, and 235 were study specific. A core of 30 frequently cited tools was identified. The use of previously published assessment tools to evaluate primary study outcomes was associated with higher PEDro scores. Significant heterogeneity in outcome assessment was demonstrated, although a core of 30 frequently cited tools could be identified. Appropriate evaluation and selection of outcome measures would enhance the methodological quality of randomized controlled trials. PMID- 17912136 TI - Joint angle and contraction mode influence quadriceps motor neuron pool excitability. AB - OBJECTIVE: The purpose of the study was to compare the central activation ratio (CAR) of eccentric contractions to isometric contractions at 30 and 70 degrees of knee flexion. DESIGN: A repeated-measures design was used. CARs were measured at 30 and 70 degrees of knee flexion in 16 healthy subjects during both eccentric and isometric modes of contraction. CARs were measured using the superimposed burst technique. RESULTS: Isometric CARs at 30 degrees (0.88+/-0.069) of knee flexion were significantly higher (P<0.001) than at 70 degrees (0.77+/-0.116). Eccentric CARs were significantly higher (P=0.013) at 70 degrees (0.87+/-0.085) of knee flexion compared with 30 degrees (0.8+/-0.09). At 30 degrees of knee flexion, isometric CARs were significantly higher (P=0.003) than eccentric CARs. At 70 degrees, eccentric CARs were higher (P<0.001) when compared with isometric CARs. CONCLUSIONS: Our results provide evidence that isometric measures at a single joint angle are not sufficient in generalizing activation of an entire muscle group for dynamic movements. CARs are significantly affected by joint angle and mode of contraction. PMID- 17912138 TI - A dynamic seating intervention for wheelchair seating discomfort. AB - OBJECTIVE: The objective of this study was to examine the effectiveness of a new user-adjustable wheelchair seating system designed to relieve discomfort for long duration wheelchair users. DESIGN: This objective was carried out using the newly developed Tool for Assessing Wheelchair disComfort (TAWC) as the primary outcome measure. Two wheelchair users each tested two different designs and feedback from the wheelchair users regarding the first design was used to guide development of the second design. A single-subject research methodology was used, allowing long duration (up to 2 wks per test) evaluation of the wheelchair seating systems and comparison of subject discomfort levels with those experienced during a baseline period using their own wheelchairs. The experimental wheelchair seating systems employed existing automotive seating with embedded pneumatic bladders that allowed adjustment of the seat and back-support characteristics. The test wheelchair also had tilt, recline, and elevating leg rests. RESULTS: The two subjects completed limited periods of testing with the first design, both finding poor results with either stable or increased levels of discomfort. Subject feedback was used to redesign the wheelchair seat. After redesign, both subjects tested the second design and found it substantially more comfortable. CONCLUSIONS: The selected research methodology was a very positive method for a progressive wheelchair seating design and the second design provided improved comfort for both users when compared with that experienced using their own wheelchairs and the first test wheelchair. Future research of this type of user controlled technology is recommended. PMID- 17912139 TI - Six-minute walk test as an outcome measure: are two six-minute walk tests necessary immediately after pulmonary rehabilitation and at three-month follow up? AB - A 3-mo prospective, longitudinal, repeated-measures study was undertaken in subjects with chronic obstructive pulmonary disease (COPD). The study aimed to determine whether there was a difference in 6-min walk distance (6MWD) when two 6 min walk tests were performed after pulmonary rehabilitation (n = 44) and at 3-mo follow-up (n = 40), and whether the results reflected the program outcomes. There was a significant increase in 6MWD between two 6-min walk tests before rehabilitation (P < 0.001), and at 3-mo follow-up (P < 0.001), but not immediately after rehabilitation (P = 0.1). In terms of program outcomes, there was an increase in 6MWD from before to after pulmonary rehabilitation (P < 0.001); however, the increase was greater if the better of two tests was reported. Six-minute walk tests performed twice before and after pulmonary rehabilitation programs and at 3-mo follow-up assessments ensure accuracy of measurement of the 6MWD and program outcomes. PMID- 17912140 TI - Use of weekly alendronate to treat osteoporosis in boys with muscular dystrophy. AB - Three boys with muscular dystrophy with known osteoporosis were each treated for 1 yr with weekly alendronate and daily calcium and vitamin D. Measurements of lumbar spine and proximal femur using dual-energy x-ray absorptiometry were obtained at the initiation of the alendronate, at 6 mos, and at 1 yr. All three boys demonstrated increases in bone mineral density, with z scores improving from baseline to 1-yr follow-up. Improvements were observed at the lumbar spine, femoral neck, and greater trochanter. In this small case series, weekly oral alendronate for 1 yr plus daily vitamin D and calcium was effective in improving bone mineral density. PMID- 17912141 TI - Kinematic analysis of hand movements after tendon repair surgery: a new assessment using drawing movements. AB - OBJECTIVE: Although several hand outcome tests exist to judge skill level after hand injury, currently none give insight into how tasks are performed by looking at kinematic parameters. In this article, the clinical value of analyzing kinematic parameters related to the drawing of a triangle on a graphics tablet by healthy subjects and patients with hand injury is discussed. DESIGN: In a first experiment 10 healthy subjects drew the triangles as accurately as possible at various speeds. In a second experiment, 67 healthy subjects and 12 patients with flexor tendon injury were measured repeatedly. RESULTS: In the first experiment, the analysis showed a high linear correlation between speed and accuracy for each individual (Pearson correlation coefficient >/=0.762, P 50%) in one patient, in whom coronary intervention was performed subsequently. CONCLUSION: The paclitaxel-eluting stent Taxcor appeared to be effective and safe up to 6 months following implantation. PMID- 17912162 TI - Antithrombotic therapy and the transition to the catheterization laboratory in UA/NSTEMI. AB - The management of unstable angina/non ST elevation myocardial infarction (UA/NSTEMI) has evolved substantially in recent years. Multiple new antithrombotic options are available; in addition, the use of interventional strategies in patients with UA/NSTEMI has become the dominant strategy, particularly in tertiary centers. On the one hand, we are doing more percutaneous interventions more rapidly in ACS patients. On the other hand, we have an ever expanding therapeutic armamentarium to apply in these complex clinical circumstances. Much of the controversy surrounding modern-day management is not so much about the specific the choice of agent or strategy, but rather how to use these agents most effectively in a clinical environment where patients may come forward to the catheterization laboratory, sometimes rapidly, and may require percutaneous or surgical revascularization. All available antithrombotic agents act on one (or more) of the four steps of coagulation: platelet activation, platelet aggregation, thrombin generation, and thrombin activity. The antiplatelet agents, aspirin, thieno-pyridines, and glycoprotein (GP) IIb/IIIa antagonists, target the early steps of platelet activation and aggregation. The antithrombin agents, unfractionated heparin, low molecular weight (LMW) heparin, Xa inhibitors, and direct thrombin antagonists, act specifically to target thrombin generation, thrombin activity, or both. We will review the major recent trials that comprise the current state of knowledge regarding these new antithrombotic agents in ACS, and discuss some of the near-future additions to our armamentarium, including prasugrel, Cangrelor, and AZD6140. The most recent ACC/AHA and ESC unstable angina guidelines have emphasized that multiple options are available, and no one agent can be recommended over the others in all cases. There is NOT one perfect antithrombotic regimen for all patients. Antithrombotic therapy needs to be individualized, and that so-called ''standard'' therapy may need to be supplemented (or even replaced) in specific circumstances. Ultimately, determining optimal therapy means understanding the physiology, understanding the therapeutic options - not just how they work, but how they may work together, and being able to interpret a never-ending supply of new clinical trial data that have to be applied in the ''real world''. PMID- 17912163 TI - Revascularization in high risk patients: diabetes mellitus. AB - Coronary artery disease is the main cause of morbidity and mortality in patients affected by diabetes mellitus. Pathophysiology of atherosclerosis in diabetics exhibit specific characteristics that confer them a high risk. In this regard, revascularization in diabetic patients remains a challenge. Both techniques [percutaneous coronary interventions (PCI) and coronary artery bypass graft (CABG)] demonstrate poorer outcomes in diabetics as compared to non diabetic patients. When one revascularization modality has been compared against the other, CABG has consistently demonstrated to be more efficacious than PCI. Thus, current guidelines recommend CABG for the treatment of multivessel disease in diabetics. However, the efficacy of recent developments in the PCI field (i.e. drug-eluting stent) is currently under investigation. The aim of this review is to update current evidence in the field of coronary revascularization in diabetics. Evolution of PCI over time will be specifically addressed as well as current evidence of drug-eluting stents in terms of efficacy and safety in diabetics. Besides, main CABG vs PCI trials will be reviewed. Additionally, we will focus on potential complications to be faced in either revascularization modality. Finally, revascularization in specific subgroup of diabetic patients such as those presenting with acute coronary syndromes or those with diabetes related systemic complications will also be addressed. PMID- 17912164 TI - Revascularization in the high-risk patient: multivessel disease. AB - The aim of this article is to review the treatment of patients with multi-vessel coronary artery disease. Percutaneous coronary intervention (PCI) has been challenging coronary artery bypass grafting (CABG) as the gold standard of care for patients with multi-vessel disease; however, the application of PCI to these patients has been mainly limited by restenosis. Up to the late 1990s, numerous large-scale, randomized trials addressed this issue comparing CABG to PCI with balloon angioplasty or bare-metal stents. These studies demonstrated similar rates of death and myocardial infarction in both groups, while the need for revascularization remained significantly lower in the CABG group. Drug-eluting stents (DES) have dramatically reduced restenosis and repeat revascularization rates. CABG has also progressed with improvements in perioperative management, a higher use of arterial grafting, and advanced techniques with the implementation of minimally invasive and off-pump surgery as options. Therefore, the results of previous trials in the pre-DES era can no longer be extrapolated into the ''real world''. As intermediate steps preceding a fully-fledged, randomized trial, several trials have compared PCI with DES and the historical control of CABG, but the results are still inconclusive. Several dedicated randomized trials are currently ongoing to compare PCI with DES and CABG using contemporary techniques. Until the results of these randomized trials are presented, the choice for each strategy should be based on the patients' individual risk and anatomy. PMID- 17912165 TI - Emergency percutaneous coronary intervention (PCI) for the care of patients with ST-elevation myocardial infarction (STEMI). AB - There is general consensus that emergency percutaneous coronary intervention (PCI) is the preferred treatment for patients with ST-elevation myocardial infarction (STEMI), so long as it can be delivered in a timely fashion, by an experienced' operator and cardiac catheterization laboratory (CCL) team. STEMI is both a functional and structural issue. Although it has been recognized since the work of pioneering cardiologists and surgeons in Spokane, Washington, that approximately 88% of patients presenting within 6 hours of onset of STEMI have an occluded coronary artery, it is the pathophysiology of myocardial necrosis, and the varied consequences of necrosis that characterize STEMI. Accordingly, experience' of both primary operator and cardiac catheterization laboratory (CCL) crew, in performing an emergency PCI for STEMI, are as much a function of experience with the treatment of complex MI patients, as experience with coronary intervention. Rapidly achieving normal coronary artery flow, at both the macro and micro vascular levels, is the recognized key to aborting the otherwise progressive wavefront' of myocardial necrosis. The time urgency of decisions (Time is muscle') make emergency PCI for patients with on-going necrosis, more like emergency room (ER) care, than like most in-hospital or outpatient care. In general, most patients with acute coronary syndromes (ACS) are currently thought to have plaque rupture and/or erosion with subsequent thrombosis and embolization. Consequences of thrombo-embolism, such as slow flow' or no-reflow' are in addition to, the structural (anatomic) considerations of PCI in stable patients (such as ostial location; bifurcation involvement; heavy calcification; tortuosity of lesion or access to it; length of disease; caliber of infarct artery; etc.). Good quality studies have provided strong support for the specific added value of glycoprotein IIb/IIIa inhibitors (especially abciximab), dual antiplatelet therapy (the addition of the thienopyridine, clopidogrel, to aspirin use), and bare-metal stents (BMS), for a broad range of STEMI patients. The added value of drug-eluting stents (DES) to bare-metal stents (BMS), primarily in terms of reducing restenosis and repeat revascularization, is supported by several randomized trials, and a number of registries, despite its being off-label' from a regulatory standpoint. The recognition of late stent thrombosis (LST) has raised additional issues, in choosing between these two options for specific STEMI patients. The added value of a number of other mechanical approaches to coronary thrombus, such as thrombus removal devices, and/or distal protection, are more controversial, and perhaps, patient specific. Whether intravascular ultrasound guidance (IVUS) for stent use should be used for the majority, or even a specific minority, of STEMI patients, is also controversial; late-stent thrombosis provides a counter-point. The advantages of developing a network approach to STEMI care, so as to optimize the number of patients receiving timely reperfusion, have been demonstrated in Prague, Denmark, and Minneapolis, among many places. The benefits of both bivalirudin (anti-thrombin drug with efficacy against clot-bound thrombin, which does not appear to stimulate platelets) and abciximab (glycoprotein IIb/IIIa inhibitor which is antibody to platelet receptors), as PCI adjuncts generally, and for STEMI patients, in particular, are supported by multiple trials. The specific choice of administering the bolus dose of either, or both, drugs via intra-coronary (IC) injection follows the precedents' of IC thrombolytics, and IC small-vessel vasodilators for no-reflow', but it has not been tested by prospective, randomized trials. Although rapid reperfusion is the first objective, one cannot ignore the other components of the oxygen delivery chain, and the importance of each of these components to on-going delivery of oxygen to all vital organs. A balance must be struck between doing those control' things which serve to stabilize other vital components of the oxygen-delivery chain, without digressing too long from the job of re establishing brisk coronary flow. The clinical and angiographic heterogeneity of STEMI patients and the array of available therapeutic approaches make it impossible to obtain specific randomized trial direction for many of the clinical decisions in an individual emergency PCI for STEMI. There are a range of reasonable/ appropriate therapeutic choices for a given emergent PCI performed by multiple experienced and competent operators. The treatment of STEMI, and high risk non-STEMI, patients, by means of emergent PCI, is among the most challenging and rewarding arenas in contemporary medicine. PMID- 17912166 TI - New pharmacologic options in the treatment of acute coronary syndromes and myocardial ischemia-reperfusion injury: potential role of levosimendan. AB - Modern and effective therapeutic possibilities have improved the management and outcomes in acute coronary syndromes and acute myocardial infarction. However, substantial morbidity and mortality still remain. Myocardial ischemia-reperfusion injury may contribute to additional damage to myocardial necrosis and apoptosis. Therefore, it has been focused on attention and field of therapeutic actions in the last years. The main mechanisms involved in the pathogenesis of ischemia reperfusion injury are depressed energy metabolism, elevated oxidative damage, and altered calcium homeostasis. In experimental trials, a variety of drugs have proved effectiveness for the prevention and treatment of the ischemia-reperfusion injury. However, its efficacy, not always confirmed, has not yet been established in clinical practice. On the basis of the strong evidence linking potassium ATP dependent channels opening in the myocardium and its proved cardioprotective role during ischemia, these channels have been pointed out as possible and promising pharmacological targets in this setting. Some evidences suggest that the calcium sensitizing agent levosimendan may have of beneficial and exerts cardioprotective effects on myocardial ischemia and ischemia-reperfusion injury. Further investigation is warranted on this novel application of levosimendan. PMID- 17912167 TI - Percutaneous treatment of diseased saphenous vein grafts: current state-of-the art and future directions. AB - The percutaneous treatment of patients with obstructive atherosclerotic disease in diseased coronary saphenous vein bypass grafts still remains a challenge in interventional cardiology. We discuss the actual evidence-based knowledge for the percutaneous management of this lesion subset, focusing on the devices that are actually considered the gold standard for this treatment: bare-metal stents and distal protection devices. We also comment the negative results of the trials regarding the promising covered stent-grafts. We finally offer insights into the currently available evidence for the use of drug-eluting stents in saphenous vein grafts. These devices are potentially a promise for the successful sealing of vein graft disease, however, available long-term safety and effectiveness data are conflicting and give reason for caution. PMID- 17912168 TI - Multislice computed tomography coronary angiography: clinical applications. AB - Cardiac and coronary computed tomography (CT) is becoming increasingly common in clinical practice. Even if there is no well-established evidence, this diagnostic modality is so strong and effective and, in skilled hand, it can be readily used in clinical practice. After learning its potential and the technical limits, this tool could be used for risk stratification as well as for revascularization evaluation. In this review, we will describe the results of present literature, clinical applications at present considered suitable to CT technology (i.e. 64 slice and dual-source scanners) and future applications and innovations. PMID- 17912169 TI - Bone marrow stem cell therapy for cardiac repair: challenges and perspectives. AB - Bone marrow (BM) stem cells can differentiate into multiple cell types, including vascular cells and, possibly, cardiac myocytes. Stem and progenitor cells are mobilized into the peripheral circulation early after myocardial infarction. Experimental evidence suggests that BM-derived cells injected into infarcted hearts can improve cardiac function. However, mechanisms underlying functional improvements remain unclear. Initial randomized, placebo-controlled trials in patients with acute myocardial infarction have provided controversial RESULTS: On the one hand, a modest but significant and sustained improvement in left ventricular function was observed in the Reinfusion of Enriched Progenitor Cells and Infarct Remodeling in Acute Myocardial Infarction (REPAIR-AMI) study contributing to the better clinical course. Results of other studies were neutral. Differences in the study design, cell processing or timing of cell delivery might explain, in part, different outcomes among studies. Furthermore, studies in patients with chronic ischemic heart disease remain observational, and therapeutic effects using surrogate end-points needs to be demonstrated. Thus, there is a need for further coordinated research with well designed, hypothesis driven clinical trials, in parallel with fundamental research aimed at understanding the mechanisms underlying the biological and functional effects of BM cell therapy for cardiac repair. PMID- 17912170 TI - Endovascular intervention in the treatment of congenital heart disease in adults. AB - Over the last years, endovascular intervention have become an important part of treatment in patients with congenital heart disease particularly for residual defects after surgery done in infancy. These transcatheter procedures can be described as dilatation of stenotic sites (angioplasty, endovascular stenting and valvuloplasty) or as a closure of anomalous openings (device closure defects and vascular embolisation). Balloon valvuloplasty, without or with stent, is the procedure of choice in adults with pulmonary valve stenosis, pulmonary arteries stenosis, bicuspid aortic valve stenosis without calcification, aortic re coarctation. Treatment of native aortic coarctation is still under debate. Devices for closing atrial and ventricular septal defects or patent ductus arteriosus have been developed and are now widely used. Transcatheter, plug or coil occlusion is nowadays the goal treatment in a wide range of arterial and venous vascular connections. This review describes the current role of each major catheter-directed therapy in the treatment of congenital heart disease in adults. PMID- 17912171 TI - Percutaneous closure of the patent foramen ovale. AB - A patent foramen ovale (PFO) is a common finding present in 25% of the population. A relationship between PFO and several clinical conditions such as stroke, migraine, platypnea-orthodeoxia syndrome, neurological decompression illness in divers, high altitude pulmonary edema, sleep apnea, and economy class syndrome have been documented. Observational non-randomized studies have shown percutaneous PFO closure more effective than medical treatment for stroke prevention, in particular in patients with complete closure as well as in patients with more than one cerebrovascular event at baseline. In the case of migraine, PFO closure has been shown to result in a marked reduction in migraine burden or migraine days. PFO anatomy, epidemiological data on associated clinical conditions, comparison between percutaneous closure and medical treatment, as well as the technical aspect of the procedure are described in this review. PMID- 17912172 TI - Transcatheter closure of postinfarction ventricular septal defects. AB - Postinfarction ventricular septal defect (VSD) is a rare but fatal complication after myocardial infarction. Surgery for postinfarction VSD is considered the gold standard. However, it is associated with high morbidity and mortality, patient discomfort, need of cardiopulmonary bypass, sternotomy, and skin scarring. As a consequence, less invasive interventional techniques have been developed for postinfarction VSD closure. The current review focuses on patient selection, techniques, results, potential complications and outcome of percutaneous postinfarction closure. PMID- 17912173 TI - [Association of antioxidants and natural immune activators in the treatment of astheno-teratospermia and abacterial leukocytosis]. AB - AIM: Leukocytes are often present in human seminal plasma and more frequently in infertile men. Leukocytospermia is associated with sperm morphological and functional alterations. Immune cell activation leads to an increase of free radical production, without any antioxidant defence activation. Leukocyte presence during sperm maturation and migration through male genital tract and consequently exposure to reactive oxygen species led to sperm alteration: axonemal, acrosomal and nuclear structure damage, associated with necrosis. In order to evaluate the immune-modulating and antioxidative activity of beta glucan, fermented papaya and lactoferrin associated with vitamins C and E, we analysed sperm characteristics of selected infertile male with astheno teratospermia and abacterial leukocytosis. METHODS: We selected 20 patients referred to our Sterility Centre for semen analysis with leukocyte concentration higher than 1x106 cell/mL. Seminal quality evaluation was performed according to WHO guidelines (1999) using Papanicolau and eosin staining, before and after three months of treatment with beta-glucan, papaya, lactoferrin, vitamin C and E. RESULTS: After therapy, seminal analysis showed a significant reduction of leukocyte concentration and an increase of sperm motility and normal sperm morphology. CONCLUSION: Our results suggest that a combined immunomodulating and antioxidant treatment protect sperm cells during maturation and migration through the male genital tract, resulting in a functional rescue demonstrated by the improvement of semen quality. PMID- 17912174 TI - Clinical practice evaluation of combination of atosiban, ritodrine and ketoprofen for inhibiting preterm labor. AB - AIM: The aim of this study was to compare the efficacy and tolerability of atosiban vs ritodrine administered as single-drug or as combination therapy with the COX inhibitor ketoprofen in the treatment of preterm labor and to investigate how frequent is the need for combination therapy with ketoprofen. METHODS: Ninety one women with diagnosis of threatened preterm delivery at 24-33 weeks' gestation were enrolled in an observational case-control study. Forty-seven received IV atosiban (6.75 mg initial dose, 300 microg/min loading dose for 3 hours, 100 microg/min maintenance dose for 48-96 hours) and 44 IV ritodrine (0.05-0.3 mg/min). When response to the first drug in the first 2-4 hours was unsatisfactory, ketoprofen was added (100 mg loading dose IV and 100-150 mg maintenance dose every 12 hours) for a maximum of 48 hours. RESULTS: Ketoprofen was added in 51.1% of the atosiban group and 47.7% of the ritodrine group (P 0.75, not statistically significant). The percentages of women non delivered in the two groups were 85.1% vs 81.8% at 48 hours (P=0.44) and 59.6% vs 54.5% at 7 days (P=0.39). One woman treated with atosiban reported transient dyspnea at the administration of the bolus dose; 20.5% of women who received ritodrine developed tachycardia and 4.5% dyspnea (P=0.001). Neonatal mortality and morbidity were comparable in both groups and unrelated to ketoprofen exposure. CONCLUSION: Atosiban efficacy was comparable to ritodrine, but with a superior safety profile. A large proportion of women in both groups required second-line ketoprofen therapy, with comparable neonatal outcomes. PMID- 17912175 TI - Postpartum urinary stress incontinence: analysis of the associated risk factors and neurophysiological tests. AB - AIM: The aim of this paper is to estimate the prevalence of postpartum urinary stress incontinence (USI) three months after vaginal delivery and to analyze the risk factors more frequently correlated with USI. Pelvic floor neurophysiology was performed to assess pudendal nerve damage in symptomatic women. METHODS: A total of 562 women were interviewed and underwent urogynecological evaluation three days after vaginal delivery. They were contacted by telephone 12 weeks later. Chart abstraction was conducted to obtain obstetrical data. Three months after delivery women presenting persistent USI were invited to return for electrophysiological tests. Univariate and logistic regression analyses were performed to reveal any significant association between USI and risk factors. RESULTS: Prevalence of postpartum USI three days after delivery was 15%, decreasing to 10.6% at follow-up three months later. Multivariate analysis of risk factors revealed that persistent USI was significantly associated with: preconception USI (P<0.05), USI developed de novo after delivery (P<0.05), family history of incontinence (P<0.05), chronic cough (P<0.05) and smoking (P<0.05). No obstetric variables were independently connected with incontinence. Neurophysiological tests revealed nerve damage in 36% of the symptomatic puerperae. CONCLUSION: Persistent postpartum incontinence is associated with several maternal and urogynecological risk factors that can help to detect women at risk for early intervention. PMID- 17912176 TI - [Endometrial ablation versus hysterectomy in women treated with tamoxifen]. AB - AIM: The aim of our study is the assessment of the importance of the endometrial ablation versus hysterectomy in patients treated with tamoxifen for previous breast cancer. METHODS: Fifty-eight outpatients in therapy with tamoxifen for 1 year were controlled in the Department of Gynaecology of the University of Naples. We have selected these patients in two groups: group A, with 28 women with abnormal uterine bleeding and endometrial thickness >8 mm and group B, with 30 normal endometrium asymptomatic women. All patient of group A and 18 of group B were treated with endometrial ablation. RESULTS: Next follow-up showed normal hysteroscopy figures in 89% of cases and 5% of cases needed a hysterectomy for new abnormal uterine bleeding and cytology. CONCLUSION: Our results show the utility of endometrial ablation especially in selected cases in therapy with tamoxifen for previous breast cancer. PMID- 17912177 TI - [Intrauterine death: experience at a tertiary centre of Italy]. AB - AIM: In order to analyse causes of stillbirths, we collected all the cases observed from January 1993 to December 2006 at the Department of Gynecological Sciences, Perinatology and Child Care, University ''La Sapienza'', Rome, Italy. METHODS: For each case, age of the patient, parity, country of origin, gestational age at the moment of stillbirth, clinical condition before pregnancy, pathologies occurred during pregnancy, possible therapies and autopsy of the fetus, have been collected. To evaluate and classify the obtained data, both the NICE (Neonatal and Intrauterine Death Classification according to Etiology) and the ReCoDe (Relevant Condition at Death) classifications have been utilised; the first one being more suitable than the second for our case series. RESULTS: Results showed that among 25892 labours, 186 were intrauterine deaths (7.2%). In 1999 we noticed a decrease in the number of labours of approx. 30%, due to a reduction in the number of inpatients available spaces. The number of stillbirths presented a slithering line until 2001, while after then a marked decrease has been observed. CONCLUSION: A high percentage of stillbirths had to be classified as ''unknown causes'' (26.9%). Additional prospective research, in order to achieve a better classification, is needed. All the new cases, should be classified using the most appropriate parameter, drawing attention to all the possible issues, and centralizing the data acquired. PMID- 17912178 TI - Adjuvant treatment for young women with early breast cancer. AB - Breast cancer rarely occurs in young women (<35 years). Several data indicate that diagnosis is associated with a worse prognosis, due to a more aggressive presentation. Although the effect of chemotherapy for premenopausal women is substantial, recent evidence suggested that patients with age <35 years with endocrine responsive tumors had a significant higher risk of recurrence than older premenopausal patients with such biological characteristics. Whereas premenopausal patients with endocrine nonresponsive disease presented similar outcome. Information from different studies of three major researching cooperative groups on breast cancer patients treated with chemotherapy alone, showed a similar interaction between the age and endocrine receptor status. Innovative treatment strategies needed and the combination of ovarian function suppression with endocrine agents (such as tamoxifen) in adjuvant therapy for endocrine receptor positive tumors could be considered. Moreover, more investigation on chemotherapy, its timing, duration, and intensity are required in the adjuvant care for endocrine nonresponsive disease. A strong emotional involvement is required to those patients approaching to adjuvant therapy, which may complicate the phase of treatment decision making. There is an urgent need for tailored clinical trials on young women with breast cancer diagnosis, to allow significant progress on adjuvant treatment of these population. PMID- 17912179 TI - The use of hormone replacement therapy in patients after breast cancer. AB - Four prospective randomized studies and at least 15 observational studies investigating hormone replacement therapy (HRT) after breast cancer are available. Only the Hormonal replacement therapy After Breast cancer: Is iT Safe (HABITS) study shows an increased risk of relapse. This is probably associated with the relatively high number of patients with HRT treatment after estrogen receptor-positive cancers as well as to the preferred use of estrogen/progestin combined preparations. As is generally known, the progestin component especially seems to be mainly responsible for the probability of increased diagnosis frequency of breast cancer. However, the patient samples in all studies investigating HRT after breast cancer are small. Therefore, HRT should only be used if alternatives, such as specific not contraindicated phyto-preparations or serotonin reuptake inhibitors, are not working. This is primarily due to forensic reasons. According to medical criteria, the data for the alternatives seem to be even more sparse, since many important questions remain unanswered, such as side effects and risks, or also regarding interactions with adjuvant hormone therapy. PMID- 17912180 TI - [Treatment of female urinary incontinence: an overview]. AB - The authors reviewed the evidence-based data available in literature, regarding the treatment of the female urinary incontinence and suggested potential therapeutic strategies to approach a woman with urinary incontinence. PMID- 17912181 TI - [The prophylactic oophorectomy. An unsolved problem]. AB - The ovarian cancer is the second cancer of the female genital organs and it is the first cause of death for gynecological oncology and the sixth cause of death in women. Prevention and early detection may reduce the mortality of ovarian cancer. Potential screening tests include pelvic exams, sonography and tumor markers measurements, although they do not guarantee a positive effect on the survival rate. In the last century, the role of prophylactic oophorectomy during benign pelvic operation has been discussed. The choice of this preventive measure raises the question of the advantages and disadvantages that a surgical menopause could cause. Finally, the role of laparoscopy in the performance of the oophorectomy versus the vaginal approach is discussed. PMID- 17912182 TI - [Mucosotropic papillomavirus genotypes in the Italian population]. PMID- 17912183 TI - Clinically significant delayed postsphincterotomy bleeding: a twelve year single center experience. AB - AIM: Bleeding following endoscopic biliary sphincterotomy (ES) is associated with morbidity. We sought to identify endoscopic findings and outcome of patients with delayed bleeding after ES. METHODS: A retrospective study was carried out in a tertiary referral center. A gastrointestinal bleeding and endoscopy database was used to identify all patients with delayed post-ES bleeding from 1994 to 2006. Early endoscopic evaluation and endoscopic hemostasis was performed. Endoscopic findings, factors associated with bleeding severity, identification of stigmata of hemorrhage, and success of endoscopic treatment were identified. RESULTS: There were 84 patients with delayed post-ES bleeding. Bleeding was classified as mild in 31 (36.9%), moderate in 29 (34.5%), and severe in 24 (28.6%). Thirteen of these 84 (15.5%) experienced bleeding at the time of initial ES. Endoscopic findings for evaluation of bleeding included active bleeding in 50 (59.5%) and stigmata of recent bleeding in 34 (40.5%) patients. Endoscopic treatment was employed in 74 (88%) patients. Monotherapy was used in 39.2% and combined therapy was used in 60.8% patients. Recurrence of bleeding was significantly more common in the severe group (P<0.001). A multivariate analysis showed that coagulopathy was a predictive factor for severe post-ES bleeding. CONCLUSION: Active bleeding or stigmata of hemorrhage occurs in the majority of patients with clinically significant delayed post-ES bleeding. Endoscopic therapy is useful in controlling bleeding in nearly all cases, though multimodal therapy and repeat endoscopic procedures may be required. Coagulopathy is a risk factor for severe bleeding; preventive measures in these patients may reduce morbidity. PMID- 17912184 TI - Pancreatic stent prophylaxis of post endoscopic retrograde cholangiopancreatography pancreatitis: spontaneous migration rates and clinical outcomes. AB - AIM: Pancreatic duct (PD) stents diminish the risk of post endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP) in high-risk patients; 3 Fr stents are reported to spontaneously migrate at a significantly higher rate than 5 Fr stents in a cohort of mostly sphincter of Oddi (SOD) patients. We sought to assess spontaneous migration rates of 5 Fr and 7 Fr stents and effectiveness in preventing PEP in a diverse group of high risk patients. METHODS: A total of 4,332 ERCP exams performed between January 2002 and August 2005 were reviewed to identify patients undergoing PD stent placement. Follow-up was obtained from electronic medical records and contact with referring MDs. Plain abdominal radiographs were used to document stent passage. RESULTS: PD stents for PEP prophylaxis were placed in 246 exams (232 patients) undergoing: PD (major or minor) sphincterotomy (84), ampullectomy (50), SOD (46), bile duct precut (35), papillary stenosis balloon dilation (9) and difficult cannulation (8). Stents placed: 218 5-Fr (140 were 3 cm long and 78 =or> 5 cm long) and 28 7-Fr (12 were 3 cm long, 16=or> 5cm long). Follow-up was available in 197 (171 5-Fr, 26 7-Fr) of 246 placements (80%). Twenty of 171 5-Fr stents were electively removed via EGD within=or< 24 h per endoscopist preference and were not included in analysis; 128 of the remaining 151 5-Fr stents (85%) spontaneously migrated by (or within) median of 8 days and 23 failed to pass and required EGD removal. Of 26 7-Fr stents one was electively removed =or< 24 h later; of the remaining 25, 15 (60%) spontaneously migrated by median of 16 days, 10 required EGD removal. The spontaneous migration rate of 5 Fr stents was: 1) significantly higher than 7 Fr stents; 2) significantly higher than the previously reported 67% passage rate of 5 Fr stents; and 3) similar to the previously reported 86% passage rate of 3 Fr stents. PEP occurred in 15% (n=36: 24 mild, 11 moderate, 1 severe). CONCLUSION: The spontaneous dislodgement rate of 5 Fr stents in patients where the indication is primarily non-SOD is approximately 85% - significantly higher than previously reported and similar to the reported rate of spontaneous dislodgement of 3 Fr stents in SOD patients; 5 Fr stents migrate spontaneously earlier and more frequently than 7 Fr stents. PMID- 17912185 TI - Inflammatory bowel disease: an overview. PMID- 17912186 TI - Pathogenesis and clinical approach to extraintestinal manifestations of inflammatory bowel disease. AB - Crohn's disease (CD) and ulcerative colitis (UC), both should be considered as systemic diseases as they are associated with clinical manifestations involving the organs outside the alimentary tract. In a genetically susceptible host with inflammatory bowel disease (IBD), complex interaction of bacterial or other local factors in the colon with antigen presenting cells may trigger an immune reaction to a shared antigen in the involved organs. These extraintestinal manifestations (EIM) are observed in up to 20-40% of the patients with IBD. Patients with CD are more susceptible to EIMs than patients with UC. Joints, eyes, skin and biliary tract are the most commonly involved organ systems. Some manifestations such as uveitis, episcleritis may precede the onset bowel disease and some may occur in conjunction with or subsequent to the diagnosis of active bowel disease. Although many EIMs tend to follow the clinical course of IBD and respond to the treatment of underlying bowel disease, some EIMs such as primary sclerosing cholangitis and ankylosing spondylitis tend to follow a course independent of the bowel disease activity. Biological agents, particularly anti-TNFa based therapies now assume an important role in the treatment of EIMs. Early recognition and treatment of EIMs are crucial in preventing major morbidity. PMID- 17912187 TI - Emerging biological treatments in inflammatory bowel diseases. AB - Although the advent of infliximab has changed the treatment paradigm and goals in inflammatory bowel diseases (IBD), it does not provide a cure for IBD and recent evidence has demonstrated that the immunogenicity of this chimeric anti tumor necrosis factor (TNF) antibody is associated with secondary loss of response and intolerance. In ulcerative colitis the efficacy of infliximab was demonstrated in two large clinical trials, but long term maintenance efficacy data are lacking. Novel biological agents have entered clinical development and pioneering trials have been reported in the last two years. For Crohn's disease the fully human IgG1 anti TNF monoclonal adalimumab, the humanized pegylated Fab-fragment certolizumab-pegol and the humanized anti a4 integrin IgG4 antibody, natalizumab have yielded the most promising results in controlled trials, but also agents inhibiting the crucial IL12/interferon-g feedback loop suggest therapeutic potential. For severe ulcerative colitis infliximab has been shown to be an effective rescue treatment and the anti T-cell CD3 antibody has shown promising open label RESULTS: Crucial in the development of novel biological agents, however, is the benefit risk ratio. As illustrated by unexpected but devastating brain infections with antiadhesion molecules, clinicians should be aware that the powerful immunomodulatory capacity of biologicals necessitates a rigorous safety follow-up. PMID- 17912188 TI - Capsule endoscopy: a comprehensive review. AB - The development of wireless capsule endoscopy (CE) has been a significant technologic advancement for the non-invasive visual evaluation of the entire small bowel and esophagus. The capsule endoscope is disposable and measures 11 x 26 mm. There are two capsule endoscopes currently available: PillCam ESO and PillCam SB for the evaluation of the esophagus and the small bowel, respectively. The PillCam ESO has two cameras on each end and captures images 7 frames per second per each camera, while the PillCam SB has one camera and captures images at a rate of 2 frames per second. Both capsule endoscopes transmit the image data using a radiofrequency signal to a recording device worn on the patient's waist. Once image acquisition is completed, the data from the recording device is downloaded to a computer workstation and analyzed by a gastroenterologist. Common indications for the small bowel CE include obscure gastrointestinal bleeding (OGIB), suspected Crohn disease (CD), as well as other suspected small bowel pathologies, while indications for the esophageal CE include screening of Barrett esophagus and esophageal varices. Small bowel CE appears to be more sensitive in the evaluation of OGIB and small bowel CD compared with other conventional radiological and endoscopic modalities. Preliminary results for the esophageal CE reveal good accuracy for screening of both Barrett and esophageal varices. CE is well tolerated by most patients, requires no sedation, and carries few side effects. One of the complications of CE is capsule retention; however, a patency capsule system has been developed, which can indicate whether an obstructing lesion is present before CE is performed. Finally, there are preliminary data suggesting that a new capsule endoscope for the colon may be useful in the evaluation of patients for colon polyps and possibly, screening for colon cancer. PMID- 17912189 TI - [Focus on folic acid benefits]. AB - Folic acid, or vitamin B9, derives its name from the Latin word folium, which was used to indicate large-leafed plants rich in folic acid. Naturally occurring in spinach leaves, this complex acid was first synthesized in 1945. Since then, mounting scientific evidence has demonstrated its beneficial effects on the human body. An essential nutrient for well-being and health, folic acid intake during pregnancy is especially important for preventing neural tube defects. This focus highlights the importance of correct intake levels of folic acid, particularly in subjects at risk of vitamin B(9) deficiency. PMID- 17912190 TI - Hepatocellular carcinoma complicating primary sclerosing cholangitis in Crohn's disease. A case report. AB - The prevalence of primary sclerosing cholangitis (PSC) in Crohn's disease (CD) patients is up to 8.5%. Although cholangiocarcinoma may complicate long-standing PSC in one third of the cases if follow-up is extended long enough, hepatocellular carcinoma (HCC) is a rare complication of PSC. The concomitant presence of PSC, HCC and CD have been reported sporadically. We discuss here a case of association of these three conditions. PMID- 17912191 TI - Recurrent pancreatitis as a manifestation of multisystem mitochondrial disorder. AB - In single cases mitochondrial disorders may manifest as pancreatitis, but recurrent, chronic pancreatitis with exacerbations of at least 15 times without morphological alterations of the pancreas but concomitant diabetes mellitus has not been reported. In a 57-year-old Caucasian male mitochondrial disorder was diagnosed at the age of 49 years upon epilepsy with generalized and focal seizures, cognitive decline, migraine, mitochondrial myopathy, polyneuropathy, diabetes mellitus, hypokalie-mia, hyperlipidemia, atrial fibrillation, heart failure, sicca syndrome, recurrent pancreatitis, chronic diarrhea, polydipsia, hyperhidrosis, steatosis hepatis, anemia, thrombopenia, an abnormal lactate stress test, and a muscle biopsy showing ragged-red muscle fibers, single completely COX-negative fibers, target fibers, increased number of sarcoplasmatic lipid droplets, but normal mitochondrial morphology on electron microscopy. Between the age of 33 years and the age of 44 years, at least 15 episodes of pancreatitis, manifesting as severe abdominal pain, and elevated exocrine pancreatic enzymes, but without morphological alterations of the pancreas, responding well to H2-blockers and food restriction had occurred. Recurrent pancreatitis without morphological alterations of the pancreas may be a feature of multisystem mitochondrial disorder resulting in diabetes mellitus. Physicians should familiarize with pancreatitis as a manifestation of a mitochondrial disorder and mitochondrial disorder should be excluded in patients with pancreatitis. PMID- 17912192 TI - Acute gastrointestinal bleeding following aortic valve replacement in a patient with Heyde's sindrome. Case report. AB - A 58-year old man was admitted to the hospital because of melena. He had a 1-year history of mechanical aortic valve replacement and coronary stent placement because of myocardial infarction and he was taking warfarin and clopidogrel. Esophagogastroduodenoscopy and colonoscopy were negative for bleeding. Capsule endoscopy showed bleeding diffuse angiodysplasia of the small bowel. The patient was treated with octreotide 20 mg, at monthly interval. After 25 months there had been no recurrence of gastrointestinal bleeding. The case suggests that mechanical valve replacement may not prevent gastrointestinal bleeding in Heyde syndrome and that octreotide treatment should be considered in these cases. PMID- 17912193 TI - Gastrointestinal involvement in Henoch-Schonlein purpura. AB - We report a 20-year-old young man with Henoch-Schonlein purpura (HSP) who presented with abdominal pain, arthralgia, and skin rash. The gastrointestinal tract was viewed by upper endoscopy and colonoscopy. We found characteristic endoscopic findings in the stomach, cecum and sigmoid colon, the combination of which has rarely been demonstrated in one patient. Histologic examination of skin biopsy specimens and gastrointestinal tract revealed leukocytoclastic vasculitis with positive staining for IgA in the capillaries. The clinical course of this patient was further improved after the combined therapy with steroids and immunosuppressive drug. PMID- 17912194 TI - It's time to define clear nutritional policies for nursing home residents in Italy. PMID- 17912195 TI - Extensive nodular lymphoid hyperplasia of the small intestine in common variable immunodeficiency. PMID- 17912196 TI - Autogenous, allogenetic and xenogenetic grafts for maxillary sinus elevation: literature review, current status and prospects. AB - Increasing patient requests for more valid prosthetic solutions usefully and aesthetically guide today's implantology to search for more reliable techniques with predictable RESULTS: Developed in the early 1980s, maxillary sinus floor elevation is a surgical technique for restoring bone tissue loss at sites for implant insertion; a variety of methods and materials are used. This article offers a review of the literature on autogenous, allogenetic and xenogenetic grafts for bone regeneration and maxillary sinus floor elevation; the focus is on the osteoconductive, osteoinductive and osteogen characteristics of the various grafts and their clinical and biological aspects. Maxillary sinus elevation with filling materials (biomaterials) is a safe oral surgery technique. Bone regeneration is an ongoing complex process leading to anatomical and functional restoration. Many events take place when biomaterials come into contact with a biological environment; molecular and cellular interactions influence tissue characteristics around biomaterials. In the presence of biomaterials, growth factors are adsorbed or wet the surface of bone substitutes, promoting graft integration with the bone. The function of biomaterials is to promote rapid bone formation; when the bone substitute is fully integrated, a gradual substitution by new bone tissue takes place. PMID- 17912197 TI - Temporomandibular disorders and bruxism. Part I. AB - Correct functioning of the entire stomatognathic system is achieved by a compatible relationship of all its parts. Four determinants, by their mutual harmonious activity, dictate the function of the entire system: the teeth, periodontium with supporting structure, muscles, temporomandibular joint (TMJ) and CNS. In such a complex system a disorder of any integrative part causes disturbances also in other parts of the system. Changed functions can arise through organic disorder, and also iatrogenically by inadequate conservative, prosthetic, surgical or orthodontic therapy. For this reason it is often difficult to recognise the primary cause. The first responses of the system to the disorder are adaptive mechanisms which occur within one or more integrative parts, and depending on their intensity and duration symptoms grow more prominent. Tissue response can be ortho- or parafunctional. Attempts are made to exclude psychoemotional influences and the obstacle eliminated by either abrasion, clenching or grinding of the teeth. If the obstacle is not eliminated by abrasion, the cause of such functional disorder becomes the trigger for parafunctional activity. From a review of the relevant literature it can be concluded that parafunctional activity is caused by changed occlusion, with determined psychological habits of the patient and specific tissue response of the stomatognathic system. Therefore, therapy of these disorders is made more difficult and includes a multidisciplinary approach. PMID- 17912198 TI - Diagnostic methods for detection of signs and symptoms of temporomandibular disorders. Part II. AB - Temporomandibular disorders (TMD) is a term which includes numerous clinical symptoms that affect the teeth and their supportive apparatus, masticatory muscles and jaw joints. They are frequently the cause of orofacial painful conditions. Etiology of dysfunction is complex and the role and importance of certain etiological factors are inadequately defined. This study includes a review of the literature and presentation of a diagnostic protocol for the detection of signs and symptoms of TMD. Optoelectronic pantography enables the application of relatively uncomplicated methods for more precise and simpler diagnosis of the above disorder, at the same time recording the results in its software and storing them, which can be extremely important during the choice of therapeutic procedure and the control of therapy efficacy. PMID- 17912199 TI - Polyostotic eosinophilic granuloma of the jaws treated by chemotherapy. A case report. AB - Eosinophilic granuloma is classified as a Langerhans cell histiocytosis. Although considered a rare pathology, up to 20% of all cases occur in the jaw bones, and radiographically may mimic odontogenic cysts or benign and malignant tumours. Different protocols have been suggested in the literature for treating eosinophilic granuloma. We report a case of polyostotic eosinophilic granuloma in a 34-year-old man showing ill defined mandibular and palatal radiolucencies. Due to the presence of multiple jaw lesions the treatment choice was chemotherapy. The clinical and radiographic feature are described, as well as treatment, complications and patient's follow-up. PMID- 17912200 TI - Economics and organ transplantation: a challenge to win. PMID- 17912201 TI - Should some Italian ICU beds be lowered from high to low level-of-care beds due to inadequate human or instrumental resources? PMID- 17912202 TI - Cost analysis of living donor liver transplantation: the first Italian economical data. AB - BACKGROUND: Over a period of 30 months, the Niguarda Ca'Granda Hospital performed 12 living donor liver transplants (LDLT) on adult subjects using the split-liver technique and transplant of the right lobe. The purpose of this work is to evaluate the financial obligation that this technique will bring, the ethical and cultural aspects, and the mortality related to surgery on a healthy donor whose only reward is in the knowledge of having done everything possible for a loved family member. METHODS: The analysis of the costs of the surgical process takes into account the simultaneous consideration of both types of patients: the donor and the recipient. The diagnostic course is subdivided into seven functional phases of the cost centers, and the transitory sequences of the foreseeable events of the entire process. The method used consists in the appraisal of all the clinical activities in chronological order several the centers of cost. The direct expenses are evaluated according to an analytical method, and the indirect costs has been carried out on the criterion of the activities of support to the process (management of the orders, recording and programming of the activities) and support to the organization (maintenance, management supplying and contests of contract, programming of the business production, management warehouses, supplyings, marketing and relations with the public). RESULTS: The cost of all the patients evaluated that were not able to donate has been added to the direct expenses of 12 donor and 12 recipient patients, in all 30 patients, so as to shift the added expenses only to the donor patient, since these costs are not included in the typical costs of transplantation from a cadaver. The indirect cost calculated for each patient has been added to the direct costs of the donor and recipient patients. The total calculated cost of LDLT is 175, 210.78 Euros. CONCLUSION: The analysis of the economical obligation that this practice brings is the starting point for an accurate evaluation of all the new technology that, in conjunction with the results of clinical efficacy and efficiency trials, is part of program of a larger scope to fulfil the general social principles of equity and justice. PMID- 17912203 TI - Proposal of a flexible structural-organizing model for the Intensive Care Units. AB - BACKGROUND: The aim of this study was to verify the capability of the Italian Group for the Evaluation of Intervention in Intensive Care Medicine (Gruppo Italiano Valutazione Interventi in Terapia Intensiva, GiViTI) Intensive Care Units (ICUs) in providing high level care (HLC) and to develop a flexible organiziational model, allowing for different levels of care in each ICU. METHODS: Once the number of active beds, personnel and technology of each ICU were determined, we computed whether the available bed number and all available resources could provide HLC according to international standards. For ICUs lacking staff or equipment for safe HLC in all declared beds, we calculated the best combination between HLC and observation/monitoring beds with less need for nurses and technology (low level of care, LLC) in order to optimise the utilization of each bed. We also investigated the work organisation of physicians and nurses in these units. RESULTS: There are 2 070 available beds in the 293 GiViTI ICUs. To provide HLC according to international criteria, the beds would decrease to 80.9%, because 144 ICUs do not have nurses or equipment to provide HLC in each bed. In order to maximize the suitable use of available resources, these ICUs would have to reduce the HLC bed number using the regained nurse workload for LLC. Because of this, the total number of HLC beds would further decrease to 65.9% of all declared beds. During Sundays and holidays, the bed/doctor and the bed/nurse ratios increase in most ICUs. CONCLUSION: To maximize the staff and equipment resources available, the bed numbers of a general ICU providing HLC must vary, even daily, according to the level of care provided. This level is not always high for all patients present. Applying this organizing model to each ICU, we could have enough flexibility to face the different demands for assistance if the ICU is built as a large open space to achieve the best clinical model and use of resources. PMID- 17912204 TI - Deformation of the 27-gauge, 3.5 inch Whitacre spinal needle: macroscopic and microscopic findings. AB - Gross deformation of the spinal needle shaft or the tip may occur during spinal needle insertion. During a one-year period, three cases of severe deformation of 27-gauge, 3.5 inch Whitacre spinal needles (B-Braun) occurred during standard spinal anesthesia in otherwise healthy patients. During this one-year period, we collected cut-bevel-point needles when bone impact occurred and compared them to the points of identical unused needles under a microscope. We believe that these microdeformations may be the cause of anatomical damage. We should be aware of even small resistance during spinal needle insertion because of the possibilities of severe deformation and injury of the anatomical structures, or of dramatic in situ breakage of the spinal needle. We should also be vigilant with respect to bone contact of the spinal needle, because needle points are damaged and can cause dural lacerations with subsequent cerebrospinal fluid leakage. PMID- 17912205 TI - Myotonia congenita and regional anaesthesia with peripheral nerve stimulation: a case report. AB - Myotonia congenita is a rare disorder caused by a defect in the skeletal muscle chloride channel function, which may cause sustained membrane depolarisation. We report a case regarding a 52-year-old man affected by this muscular pathology and scheduled for shoulder surgery that was performed under sedation and peripheral nerve block. The international literature does not elucidate clear preference toward a particular anaesthesia technique, but we believe that in this setting, regional anaesthesia and complete monitoring should be preferentially utilized when possible. In any case, the anaesthesia plan should be prudent and avoid exposure to known dangerous anaesthetics. PMID- 17912206 TI - SIAARTI - IRC recommendations for organizing responses to In-Hospital emergencies. PMID- 17912207 TI - Day Bed Unit at the Department of Gastro-enterology. Experience of a study group. AB - Day Bed Unit at the Department of Gastroenterology. Experience of a study group. We analyzed the activity of the Day-Bed Unit at the Department of Gastroenterology of Turin Hospital (Molinette). The quality of the service provided and the days of stay in hospital for each admission were evaluated in terms of cost benefit ratio. The average stay in hospital for each admission was 2.57 days as opposed to 9.3 in the in patient ward. Patients were divided in subsets according to the cause of admission. Stratifying the patients according to diagnosis showed a hospital stay of 2.73 days for liver disease and of 2.81 for pancreas and biliary disease. Among the admissions for liver disease, pre or posttranspiant patients required 2.89 days as opposed to 2.62 days of those who were treated for esophageal varices. Thus, the cause of admission was a factor influencing length of hospital stay. Patients needing non surgical treatment for liver cancer took 2.22 days if treated with percutaneous ethanol injection; those undergoing chemoembolization required 2.93 days. In conclusion, the day bed unit has proven to be able to provide a service with a good cost benefit ratio. Patients admitted to this service may be withdrawn from the waiting list of the in patient ward, thus reducing the waiting time. Optimization of this service needs integration with the surroudings Units within the Department and in the Hospital. PMID- 17912208 TI - Helicobacter pylori: a comparison of some diagnostic methods. AB - We evaluated the performances of three methods (urease-test, cytology and culture) for the detection of Helicobacter pylori infection Fifty-five patients, 28 men and 27 women (ages, 19-77 years) were included in the study. Helicobacter pylori have been detected by urease-test in 41 patients (74.5%), by culture in 39 patients (70.9%), by cytology in 32 patients (58.1%). Urease-test and culture are direct and specific methods in diagnosing Helicobacter pylori infection. PMID- 17912209 TI - Comparison between therapeutic regimes in erosive chronic gastritis associated with Helicobacter pylori. AB - Following numerous reports confirming the role of erosive chronic gastritis and ulcer, several treatment protocols have been used with the aim of eradicating this microorganism. Elimination was initially achieved using polychemotherapeutic associationsbismuth salts+metronidazole+amoxycillin and/or tetracyclineswhich proved relatively efficacious in therapeutic terms, but often led to side effects, the onset of bacterial resistance and scarse patient compliance. The authors consider that these limits can be overcome by using a new association of homeprazole+azithromycin, whose effica-cy in the eradication of Helicobacter pylo ri has been shown to be of about 85% of cases treated for a relatively short period (4 weeks). PMID- 17912210 TI - Ultrasound monitoring of alcholic liver steatosis during glutathione (GSH) treatment. AB - In addition to clinical and laboratory alterations, the ultrasound picture in patients with liver steatosis may be quite clearly and closely connected to the histological aspects of this disease. Moreover, these patients have been found to have a reduced hepatocytic and plasmatic glutathione (GSH) level and consequently a reduced liver detoxicating capacity. The aim of this study was to evaluate the therapeutic action of exogenous GSH and its positive modifying role, not only in hepatic function indices (transaminase - GT), but also in the ultrasound picture, in a group of alcholic subjects suffering from carying levels of hepatic steatosis. PMID- 17912211 TI - Laparoscopic cholecystectomy. Analysis of results of first 322 cases. AB - From April 1994 to June 1995 322 laparoscopic cholecystectomies were performed in S. Camillo hospital Treviso, 294 in election and 28 in urgency. The technique and results of this experience are reported. No mortality was observed and morbility was 1.8%. Utility of gallstones in main biliary duct screening is discussed to select patients to subject to preoperative ERCP and the role of intraoperative cholangiography. Laparoscopic cholecystectomy is considered the choice option in cholelitiasis. In case of evidence of choledocholithiasis with anamnesis or preoperative tests, the tactics of choice in authors opinion is the sequential one, providing preoperative ERCP and laparoscopic cholecystectomy. PMID- 17912212 TI - Correlation between digestive disease and psychiatric disease: casual or causal implication? AB - The aim of this study was to evaluate the prevalence of psychiatric disturbances among patients affected with digestive diseases (both organic and functional) and, viceversa, the prevalence of digestive disturbances among patients with psychiatric diseases. We performed a trasversal study on: 100 patients with organic digestive diseases and 100 patients with functional digestive diseases afferent from a Gastroenterologic Ambulatory (gastroenterologic group); 50 patients afferent from a Psychiatry Service (psychiatric group) and 50 patients afferent from a General Medicine Ambulatory affected with a non gastroenterologic active problem (control group). Each patient underwent an anamnestic, laboratory and instrumental evaluation, in order to ascertain or exclude the presence of digestive symptoms and their eventual organic basis; moreover, a semistructured interview was performed aimed at identifying a psychiatric disturbance, according to DSM-IIIr criteria. Our results showed a significantly higher prevalence: 1) of psychiatric disturbances, in the gastroentorologic group versus the control group (p<0.001), especially of somatoform (p<0.05) and anxious (p<0.001) disorders; 2) of psychiatric disturbances among patients affected by functional digestive disorders versus patients affected by organic digestive disorders; 3) of gastroenterologic disorders, in the psychiatric group versus the control group (p<0.001), with a significantly higher prevalence of functional gastroenterologic syndromes in comparison the organic ones (p<0.001). The well-established bidirectional correlation between digestive functional and psychiatric disorders is a necessary but not sufficient condition to state a relationship of direct causality between the two syndromes; however we can hypothesize that the well documented neuro-hormonal alterations may cause, on clinical grounds different symptoms, that are differently interpreted by the different specialists (gastroenterologists or psychiatrists) consulted. PMID- 17912213 TI - Drug-induced cholestasis. Clinical contribution. AB - We described two cases of drug-induced hepatitis: in both cases we recognised a pathogenesis based on hypersensivity mechanism. The first case, determined by naproxen, presented a serious cholestatic jaundice and histologic pattern due to ductular cholestasis without any inflammatory reaction. The second case, by a metildopa, was characterized by spread necrosis of liver cells and peri-portal flogistic infiltrated. This chapter of modern epatology is becoming more and more important, because of both the opportunity of studyng physiopathologica mechanism and the increasing morbidity. The peculiarity of these cases was due not only to the morbidity, but also to the seriousnes of clinical symptoms which involved problems of early differential diagnosis and of attentive follow-up. PMID- 17912214 TI - Adrenal location of the echinococcus. Considerations on a clinical case. AB - Adrenal location of a hydatic cyst is a clinical occurrence of exceptional rarity, especially in its primitive form and it could be evaluated, on the operative and autoptic comparison, not to exceed overall 0.5% of all the possible locations. Moreover, among all the cystic formations of the adrenal glands, parasitic ones are only 7%. Our observation of a case of adrenal hydatidosis, initially suspected as hepatic mass, was casually discovered by means of abdominal ultrasonography and later confirmed with the operation. This work contributes to the knowledge of this pathology, emphasizing diagnostic difficulties, and, considering the exiguous literature, helping us in supplying real indications to the surgical treatment. PMID- 17912215 TI - Pancreatic somatostatinoma presenting with chronic intestinal pseudo-obstruction syndrome. AB - A case of pancreatic somatostatinoma in a patient with long-standing symptoms of intestinal pseudo-obstruction is reported. Diagnosis was made incidentally following an exploratory laparotomy for an episode of acute intestinal obstruction. After surgical excision of the endocrine neoplastic tissue there was a decrease in plasma somatostatin levels, associated with a striking improvement of symptoms related to gut motility dysfunction. Delayed intestinal transit and gastric emptying and impaired gallbladder contraction persisted after surgery during a follow-up of 9 years in association with a positive calcium-pentagastrin test, suggestive of residual tumoral somatostatin secretion. This case report shows that a chronic intestinal pseudo-obstruction syndrome can be associated with a somatostatin-producing endocrine neoplasm and confirms the clinical heterogeneity of somatostatinoma patients. PMID- 17912216 TI - Gastrointestinal tubercular arteritis during HIV infection. AB - Although digestive tract is often involved in HIV disease, severe gastrointestinal bleeding rarely occurs. We describe a case of severe gastrointestinal bleeding associated with a mycobacterium tuberculosis arteritis of colonic tract. It is reported a morphology change of mesenteric arteries by mycobacterium tuberculosis revealed by angiography. The superselective embolization of branches mesenteric inferior artery resolved the important bleeding without surgical intervention. In AIDS patients, during intestinal tuberculosis, massive enteric-hemorrhage suggests for involvement of mesenteric arteries by mycobacterium tuberculosis. An angiography is basic to discover bleeding and if necessary for the therapeutic treatment. PMID- 17912217 TI - A new therapeutic strategy for the eradication of Helicobacter pylori infection. AB - For the eradication of the Helicobacter pylori infection, authors have tested 50 HP-positive subjects (28 females and 22 males; mean age 47 years): 21 duodenal ulcers and 29 gastritis. All patients received the following treatment: omeprazole 40 mg for 30 days, azithromycin 500 mg in a single daily dose for 3 days for 2 cycles and metronidazole 250 mgx4 for 14 days. One month after the end of therapy, patients have been controlled: the HP eradication rates have been 76% (35/46), duodenal ulcer was cured in 90% (18/20). No important side-effects were reported by patients during the treatment. In conclusion the new therapeutic scheme with azithromycin represents a semplificated alternative treatment in the eradication of Helicobacter pylori. PMID- 17912218 TI - Use of the ''Polymerase Chain Reaction'' for the diagnosis of Helicobacter pylori infection. AB - PURPOSE: H. pylori infection can be diagnosed by means of non-invasive tests or invasive techniques using endoscopy. The choice of the test depends on available instruments, type of diseases, aim of diagnostic research (therapeutic or epidemiological) and test features. PCR is able to reveal pathogenic germs in biological material with very high sensitivity and specificity. In vitro DNA amplification method consists of hybriding denaturated DNA by means of two oligonucleotide primers that allow to copy DNA fragment. The aim of our study was to determine, using PER, H. pylori colonization in the gastric mucosa of 18 consecutive patients under-went gastroscopy. MATERIALS AND METHODS: Eighteen patients complaining of dyspeptic symptoms and referred to us for upper GI endoscopy participated in the study. The studied population comprised 9 males and 9 females with mean age of 55.4 yrs (range 26-73 years). All patients underwent gastroscopy during which 4 biopsies from the antrum and 4 from the corpus were obtained for Giemsa stain, PCR analysis and histologic examination. A pair of synthetic oligonucleotides for H. pylori urease A gene, designated as HPU1 and HPU2, were used. Urease A gene fragment amplified by PCR was analyzed by 1.5 agarose gel electrophoresis. Positivity for H. pylory corresponded to PCR DNA products migrating at 411 bp after staining with ethidium bromide. RESULTS: The patients were divided into two groups, according to H. pylori infection, determined by means of Giemsa stain: group A, comprising 11 H. pylori-positive patients; and group B, with 7 H. pylori-negative patients. Our PCR assay of gastric mucosa samples proved positive in 7 cases of group A (63.6%), whereas it always proved negative among group B subjects (100%). CONCLUSIONS: Our findings, apparently in contrast with the high sensitivity of PCR, may be attributed to the lower specificity of histology or, alternatively, the absence of H. pylori in the samples tested by PCR due to the patchy distribution of H. pylori colonization in the gastric mucosa. These observations are in agreement with those from other investigations. PMID- 17912219 TI - Percutaneous treatment of non-functioning arteriovenous fistulas. AB - AIM: To show our results in the percutaneous treatment of non-functioning arteriovenous fistulas of patients undergoing haemodialysis and to discuss the results of the literature. METHODS: Thirty-four patients with 37 non-functioning arteriovenous fistulas (29 native, 8 grafts) underwent percutaneous angioplasty. Restoration of vessel lumen and function at the following dialysis were considered as successful outcome. Primary and secondary patency rates at 12 months were calculated by means of Kaplan-Meier analysis. RESULTS: Fifty-six interventions were performed, 42 on native fistulas and 14 on grafts. Immediate technical and clinical success were obtained in 98% of procedures. For native fistulas, primary and secondary patency rates at 12 months were 41% and 62%, respectively. For grafts, 14% and 34%, respectively. Only one complication was observed. CONCLUSION: Minimally invasive percutaneous treatment allows to restore function of most arterio-venous fistulas with good long term patency. PMID- 17912220 TI - The role of autologous antebrachial arteriovenous fistula in the strategy of access planning for chronic uremic patients on hemodialysis. AB - AIM: The number of arteriovenous (AV) fistula creation increases worldwide. Haemodialysis is more effective, patients live longer, and they need more access operations. The optimal strategy for the order and sequence of the different type and localization of AV fistulas remains obscure. Based on internationally acclaimed guidelines, autogenous access should be performed whenever possible and the first operation of choice is the radiocephalic fistula at the wrist, the second type is the elbow fistula. The area between the standard exposures means also good access area and its usage is not emphasized properly. Our aim was to study the short and long-term the results of autologous forearm fistulas. METHODS: Between 1997 and 2005 we performed 1018 AV shunts in an academic tertiary care centre. Ninety-seven autologous antebrachial AV shunts were performed. The average follow-up time was 31.3 months. We examined the patency rate and its connection with different variables such as diabetes mellitus, acute or chronic operative situations, indications for surgery, diameter and quality of the vein. RESULTS: The primary patency rates were 93%, 79.5% and 61.2% at the end of years 1, 2 and 6, respectively. The patency rate was not significantly affected by any of the examined variables mentioned above. CONCLUSION: The patency rate of the autologous antebrachial AV shunt is comparable to the wrist and elbow fistulas, so our results support the practice of performing fistula at this atypical localization. Proximal autologous fistulas and prosthetic graft implantation could be postponed, this way valuable time could be saved for the uremic patients. PMID- 17912221 TI - Vascular access 2007. AB - Over the past decades, vascular access management has undergone profound changes resulting in marked improvements in patient care. AV fistulae remain the access of choice and continue to represent the majority of accesses in most countries. Access monitoring is more widely available and new percutaneous therapies have revolutionized the therapeutic approach to access failure. Outpatient access centers providing both percutaneous and surgical therapies have decreased the need for vascular access related hospitalization. These advances have been supported by the development and promulgation of standardized evidence-based guidelines. These guidelines supported by national and international outcome data have helped rationalize vascular access care. Despite these advances many challenges remain. Catheter use has increased on a worldwide basis, with resultant increases in catheter related infections and complications including sepsis, endocarditis and paravertebral abscess. The availability and use of standardized chronic kidney disease care including early access placement in preparation for dialysis initiation remains underutilized in most settings. Payment system and structural barriers often provide disincentives for the efficient outpatient application of available technologies and there are a few available large prospective trials to help guide therapies. As we look to the future, it is anticipated that a wide range of new technologies will continue to improve vascular access management. Enhancing our delivery systems, combined with he thoughtful application of these technologies including new biologics, materials, interventional techniques and cellular technologies offer the promise of continued improvements in patients vascular access care over the coming decade. PMID- 17912223 TI - Choice of extracorporeal dialysis modality: can it be evidence based? AB - The incidence of end stage kidney disease is increasing worldwide and extracorporeal renal replacement techniques are widely used to treat these patients. Convective dialytic therapies such as hemodiafiltration are claimed to be superior to diffusive techniques such as hemodialysis given the higher clearance rates, hemodynamic stability and possibly reduced morbidity and mortality rates. Although observational studies have held this contention, randomized trials failed to do so. In this article, we present a case report and review available trial and systematic review evidence on the benefits-harms of various extracorporeal techniques. Both convective and diffusive clearance techniques were found to have similar all-cause mortality and hospitalization rates. Data on quality of life, dialysis related amyloidosis and procedure related outcomes such as hypotension have not been well studied. Most of the unbiased information, in the form of randomized trials, are only deriving from few and very small studies while large trials are lacking. Currently, there are three ongoing randomized clinical trials analyzing the efficacy of various extracorporeal techniques with focus on hard end points and their results will shed more light on this topic. Until then, since both convective and diffusive therapies have not been found to be different with respect to major patient-level outcomes but only some surrogates of uncertain clinical importance, the choice of renal replacement therapy should be based on other factors such as patients' preference, availability of dialysis centers and cost. PMID- 17912222 TI - Advances in peritoneal dialysis. AB - New peritoneal dialysis (PD) patients have a better survival than new haemodialysis (HD) patients in the first years on dialysis. During long-term treatment, this changes into a survival advantage for HD. The superior initial survival on PD is related to a better preservation of residual renal function of PD patients compared to HD. The importance of residual renal function is probably due to additional properties of native kidneys, such as a better removal of organic acids and low molecular weight proteins than occurs during dialysis. The magnitude of the residual glomerular filtration rate (rGFR) in PD patients is not only associated with better survival, but also with less uraemic symptoms, such as loss of appetite, and also with higher scores on quality of life tests. These relationships are absent for peritoneal clearance. Consequently measures to preserve rGFR are extremely important. Studies on an effect of peritoneal transport status on survival have given variable RESULTS: A large meta-analysis showed that fast transport patients have a 15% increased risk of death, but this is only the case for continous ambulatory peritoneal dialysis (CAPD) with conventional dialysis solutions. This suggests that the development of overhydration may be the link between transport status and mortality. A fast transport status can be inherent or acquired. The inherent form can either be due to an inflammatory state or to a large mesothelial cell mass. In both situations vasoactive mediators may be locally released. A permanent acquired fast transport status, leading to severe ultrafiltration failure develops in about one third of the patients. It is conceivable that the excess mortality in fast transport patients is caused by the types associated with an inflammatory status and with the acquired type of long-term PD. The latter, linked to morphological peritoneal alterations, is mainly caused by exposure to conventional dialysis solutions. An icodextrin based solution is especially indicated to treat ultrafiltration failure. The aim of the ''biocompatible'' solutions is to prevent the peritoneal changes. The results of animal and clinical studies are promising so far. The objective of modern PD is to extent its initial survival advantage to long-term treatment. Recent advances in knowledge of mechanisms and new dialysis solutions are likely to make this possible. PMID- 17912224 TI - New facts about encapsulating peritoneal sclerosis as a sequel of long-term peritoneal dialysis - what can we do? AB - A long lasting peritoneal dialysis (PD) leads to a special disease, so-called encapsulating peritoneal sclerosis (EPS). The hallmarks of the latter stages of the disease are intestinal obstructions and, as a consequence, malnourishment. For the precise diagnosis radiology and pathology are essential. (Triad ''typical clinical picture- typical radiology- typical pathology''.) In the middle of the pathological process of EPS is proliferative fibrosis and sclerosis of the peritoneum that subsequently leads to the assembly of the typical ''cocoon'' and obstruction. In EPS we found in the peritoneum increased amounts of vascular endothelial growth factor (VEGF) fitting the hallmark of increased neoangiogenesis and blood exudates with fibrinous matrix on the peritoneum as a feeding ground for proliferation of fibroblasts. Additionally, the number of mast cells in EPS is decreased and therefore the chymase and other fibrinolytic enzymes. The ''plasma-leak'' hypothesis focuses on fibrin and our findings help to explain most of the pathophysiology. Since the mortality of EPS is still high, emphasis should be laid on preventive treatment. Since glucose and advanced glycation endproducts (AGEs), including glucose degradation products (GDPs), are responsible for fibrosis and sclerosis of the peritoneum, biocompatible peritoneal dialysis solutions with reduced amounts of AGEs and GDPs are recommended. Additionally, a careful monitoring of patients, especially after 5-8 years of PD is very important. In case of the first signs of EPS, cessation of the modality is necessary. Thanks to this approach, most end-stage EPS pictures can be avoided. PMID- 17912225 TI - Cardiovascular disease in patients with chronic kidney disease. A clinical review. AB - Cardiovascular disease (CVD) remains the most common cause of premature death in the chronic kidney disease (CKD) population. Individuals with CKD are at 10-20 times greater risk of cardiac death than controls without CKD, despite stratification for age, race, sex and diabetes. Heightened CVD mortality begins with mild kidney disease and rises further with more advanced kidney disease. Traditional risk factors account for up to 50% of cardiovascular disease in CKD, whilst renal specific markers, including anemia, disordered bone mineral metabolism and oxidative stress, also likely contribute to the total cardiovascular burden in CKD. Despite the increased mortality, there has been a dearth of interventional cardiovascular randomized controlled trials (RCTs) in the CKD population. Furthermore, many patients with kidney disease have been excluded from the majority of mainstream cardiovascular interventional trials. While recently published RCTs on traditional and non-traditional risk factors including dyslipidemia (PPP, 4D and ALERT, VA-HIT), cardiomyopathy (FOSIDIAL, telmisartan, carvedilol), anemia (US Normal Hematocrit, CHOIR and CREATE trials), hyperhomocystenemia (ASFAST, US folic acid trial, HOST), disordered bone mineral metabolism (Cunningham meta-analysis, DCOR), oxidative stress therapy (SPACE, HOPE and ATIC, N-acetylcysteine) and multidisciplinary multiple cardiovascular risk factor intervention clinics (LANDMARK) have added to the available pool of clinical data, level 1 clinical evidence remains significantly lacking. The negative findings in many of these trials highlight the potential dangers of extrapolating findings from non kidney disease patients to those with CKD. Further large, well-designed trials are urgently required to address this issue. PMID- 17912226 TI - Dyslipidaemia in chronic kidney disease. AB - Chronic kidney disease (CKD) is associated with a highly atherogenic lipid profile, characterized by elevated triglycerides, low high-density lipoprotein (HDL) cholesterol and accumulation of small dense low-density lipoprotein (LDL) particles. Diverse mechanisms are responsible: uraemia, dialysis, immunosuppressive drugs and concomitant diseases exert their effect on the activity of key enzymes, transfer proteins and receptors involved in lipid metabolism. Post hoc analyses from large scale randomized controlled trials suggest a benefit of statin therapy with respect to cardiovascular and renal endpoints in patients with early CKD comparable to the effect in people without renal disease. Observational studies found a reduction in the risk of contrast media induced nephropathy and a reduction in the risk of hospitalization for sepsis in patients who had CKD and were treated with statins. In contrast, prospective, randomized, controlled statin trials in patients with diabetes on haemodialysis and in renal transplant recipients have not conclusively shown improvements in hard cardiovascular endpoints. This review will focus on lipid disturbances in renal disease, their impact on cardiovascular disease, existing endpoint studies and current treatment guidelines. PMID- 17912227 TI - Update on nephrolithiasis management. AB - Urolithiasis affects 10%-15% of the population in their lifetime. After spontaneous passage or surgical treatment, a subset of these patients will have recurrent calculi. These recurrent stone events are significantly morbid and can potentially lead to serious chronic renal disease, thus prevention is a very important treatment goal. Fortunately, a reversible metabolic abnormality can be identified in over 90% of recurrent stone formers. Thus, a detailed metabolic evaluation using 24 hour urine collections and serum tests is indicated in patients at high risk for stone recurrence. Once the patient's underlying urinary physicochemical and physiologic derangements are defined, targeted medical therapy can be initiated in order to prevent growth of pre-existing stones and recurrent stone formation. In this paper, we provide a review of the currently available selective and nonselective pharmacologic treatments for urolithiasis. Furthermore, we discuss a number of investigational agents for kidney stone prevention. Although many of these agents are effective, there remain numerous clinical scenarios where currently available therapies are inadequate. PMID- 17912228 TI - Acute renal failure: outcomes and risk of chronic kidney disease. AB - Acute renal failure (ARF) is a common condition, especially among the critically ill, and confers a high mortality. The incidence of ARF is increasing. Efforts such as the Acute Dialysis Quality Initiative (ADQI) are being undertaken to establish a consensus definition of ARF, and to distinguish between varying degrees of acute kidney injury that might confer a different prognosis. Data are emerging to allow comparison of the epidemiology of ARF across institutions internationally. There is ongoing recognition of the important interaction between ARF and chronic kidney disease and more information regarding recovery from ARF is available. Controversy exists regarding the optimal management of ARF. Recent publications emphasize the importance of timing and dose of renal replacement therapy rather than the modality of treatment (intermittent hemodialysis vs continuous therapies). These issues are explored in this review. PMID- 17912229 TI - A practical approach to immune monitoring in kidney transplantation. AB - In the last years, there has been a growing interest in the development of new assays available to monitor kidney transplantation. These assays will permit strategies for the minimization of immunosuppression, for induction of operational tolerance, for an early and noninvasive diagnosis of acute rejection, for prevention of adverse effects of immunosuppressive drugs and for measurement of the net immunosuppressive state. Today, there is not one test that helps us monitor the outcome of kidney recipients safely. The best strategy will be the sequential utilization of a combination of several of these tests. Further studies and further developed assays are needed to obtain an adequate immune monitoring strategy in kidney transplantation. Nowadays, pretransplant and post transplant IFN-gamma ELISPOT assay, donor-specific antibody determination, sCD30 test and Cylex(R) ImmuKnowTM assay provide valuable information about immune status that can help us guide transplant therapy in vivo. Transvivo DTH assay, tetramer staining, quantification of cell proliferation by CFSE labelling, human leukocyte antigen-G determination, phenotyping of recipient immune cells, detection of tolerogeneic dendritic cell precursors, T-cell receptor landscaping and quantification of gene and protein expression need more studies to know their exact role as monitoring markers in kidney transplant patients. PMID- 17912230 TI - Herpesviruses and polyomaviruses in renal transplantation. AB - Viral infections remain a significant cause of morbidity and mortality among transplant patients despite recent advances in early detection and treatment. Herpesviruses and polyomaviruses are the most relevant viruses post-transplant as they establish latency in immunocompetent individuals and frequently reactivate in the immunosuppressed transplant recipient. Although we have made significant strides in the early diagnosis and treatment of viral infections in renal transplant recipients over the past five years, many questions remain. Optimization of screening and prophylactic/preemptive protocols, as well as standardization of viral diagnostic testing are still needed. Understanding how viruses modify the host's immune responses, and conversely how variations between hosts' ability to mount an immune response against viruses are important areas of research that might allow for more individualization of immunosuppressive regimens. Other exciting areas of ongoing study include the associations between various HLA loci/mismatches and viral replication/infection, the mechanisms by which certain viruses (i.e., Epstein-Barr virus, human herpes virus 8) are oncogenic, and the development of new therapeutic strategies such as adoptive transfer of antigen-specific T cells to restore immunity and control viral infections. PMID- 17912231 TI - Immune monitoring after kidney transplantation. AB - During the last decennium, research in the field of organ transplantation changed from studying mechanisms of acute rejection shortly after transplantation to understanding why patients benefit from their grafts only for a limited time, because of irreversible chronic rejection and serious side effects of immunosuppression. To avoid side effects, tapering or even withdrawal of immunosuppression in transplant recipients is warranted, provided this is not accompanied with graft loss. Noninvasive cell mediated immune tests could be helpful to identify transplant recipients in whom the immunosuppressive load can be safely reduced, and to identify patients at risk for chronic rejection. In the present report, we describe cellular assays to determine donor-specific responses in peripheral blood mononuclear cells from transplant recipients taken before transplantation and during tapering or withdrawal of immunosuppression. PMID- 17912232 TI - Simultaneous kidney-pancreas transplantation. AB - Simultaneous kidney-pancreas transplantation is the most commonly performed type of pancreas transplant. Recipients with functioning pancreas transplants have normal glycemic control without the need for exogenous insulin, and are free of hypoglycemic events. While pancreas transplantation has a beneficial impact on a number of diabetic complications, and kidney-pancreas transplant prolongs survival compared to remaining on the transplant waiting list, the contribution of the pancreas to survival beyond that achieved by kidney transplant alone is controversial. Candidates generally have type 1 diabetes refractory to intensive insulin therapy; selection criteria are more stringent that for kidney transplant alone. Most pancreas transplants are performed with enteric exocrine drainage and systemic venous drainage, although portal venous drainage is also employed. Complications are more frequent and more severe than for kidney transplant alone, which is a consideration when selecting appropriate candidates. Immunosuppression usually includes induction therapy and triple-drug maintenance therapy, but early outcomes using steroid-free regimens are encouraging. Rejection is difficult to accurately detect noninvasively, but the use of percutaneous biopsy in diagnosis is increasing. Outcomes are generally good; the kidney and pancreas graft survival rates are 92% and 85%, respectively at one year. Patient survival exceeds 85% after five years. Although the benefit of the pancreas transplant on mortality is uncertain, most studies demonstrate a significant improvement in quality of life. PMID- 17912233 TI - [Uretero-vescical anastomosis during kidney transplantation. Preliminary results of two surgical techniques]. AB - AIM: The most frequent urologic complications after renal transplantation involve the uretero-vescical anastomosis (leakage, stenosis, and reflux), with a frequency of 1% to 30% in different series. METHODS: We present our results in a prospective randomized trial performed from October 2004 to September 2005, in a cohort of 36 patients, who underwent renal transplantation from cadaveric donor at our institution. A uretero-vescical anastomosis according to Lich-Gregoir was used in 18 cases (group A), whereas an anastomosis according to Knechtle was performed in other 18 patients (group B), respectively. The groups were comparable for donors and recipients characteristics. The mean donor age was 46.3 years vs 44.9 years, and the mean duration of cold ischemia was 1 086+/-296 min vs 1 100+/-381 min for group A and for group B respectively. The mean recipient age was 47.5 years vs 46.1 for group A and group B, respectively. RESULTS: No differences were evidenced between the two uretero-vescical anastomosis in term of surgical complications, infections or patient and graft survival at one year of follow-up. Stenosis and leakage involved 2 patients for each group respectively. Numbers of infections, days of antibiotic therapy were similar between the two groups. CONCLUSION: Our early experience does not evidence differences between the two types of uretero-vescical anastomosis. PMID- 17912234 TI - Modification of adenovirus gene transfer vectors with synthetic polymers: a scientific review and technical guide. AB - The chemical modification of adenovirus (Ad) gene transfer vectors with synthetic polymers is a promising strategy for overcoming typical in vivo hurdles associated with Ad-mediated gene delivery. Polymer-modified Ad vectors induce significantly reduced innate immune responses, can evade pre-existing anti-Ad antibodies, allow for repeated vector delivery, and have been used for developing novel retargeting strategies. The most widely used polymers for covalent chemical capsid surface modification are poly-N-(2-hydroxypropyl)methacrylamide (poly HPMA) and polyethylene glycol (PEG), and the latter is in wide clinical use for modifying protein biopharmaceuticals. In this review, we critically compare the properties of various polymers with respect to Ad vector shielding and retargeting, and identify areas for future research on polymer-modified viral vectors. We describe the potential technical pitfalls of polymer modification of Ad vectors and provide a technical guide for avoiding these while establishing polymer modification techniques in the laboratory. PMID- 17912235 TI - RNA aptamer-targeted inhibition of NF-kappa B suppresses non-small cell lung cancer resistance to doxorubicin. AB - Due to the prevalence of tumor chemoresistance, the clinical response of advanced non-small cell lung cancer (NSCLC) to chemotherapy is poor. We suppressed tumor resistance to doxorubicin (Dox) in A549 cells, a human NSCLC cell line, both in vitro and in vivo in a lung tumor xenograft model, using a novel adenoviral expression system to deliver an RNA aptamer (A-p50) that specifically inhibits nuclear factor-kappaB (NF-kappaB) activation. By achieving selective, targeted, and early inhibition of NF-kappaB activity, we demonstrate that NF-kappaB plays a critical role in Dox-induced chemoresistance by regulating genes involved in proliferation (Ki-67), response to DNA damage (GADD153), antiapoptosis (Bcl-XL), and pH regulation (CA9). This Dox-induced NF-kappaB activation and subsequent chemoresistance is dependent on expression of p53. We also demonstrate that NF kappaB promotes angiogenesis in the presence of Dox via the hypoxia-inducible factor-1alpha/vascular endothelial growth factor (HIF-1alpha/VEGF) pathway, revealing a previously unknown mechanism of NSCLC resistance to Dox. These studies provide important insights into the mechanisms of Dox-induced chemoresistance, and they demonstrate a novel, effective, and clinically practical strategy for interfering with these processes. PMID- 17912236 TI - Durable, safe, multi-gene lentiviral vector expression in feline trabecular meshwork. AB - Multiple disease-specific considerations have led to interest in the potential of gene therapy to permanently correct elevated intraocular pressure (IOP), the main causal risk factor for primary open angle glaucoma (POAG). Since IOP elevation results from abnormal resistance to aqueous humor outflow from the eye through the trabecular meshwork (TM), a means to genetically modify this specialized outflow organ permanently and safely is a prioritized goal. Here we tested different lentiviral vector designs and doses for long-term transgene expression in a large animal model, and investigated whether exogenously introduced myocilin proteins influenced IOP. The anterior chambers of 18 domestic cats (36 eyes) were injected with dual-gene feline immunodeficiency virus (FIV) vectors. Substantial, well-tolerated green fluorescent protein (GFP) expression was achieved in TM and monitored non-invasively in vivo for 1.2-2.3 years, using both 5' cap-translation and internal ribosome entry site (IRES)-translation. In all 36 eyes, post-mortem examination revealed substantial TM transgene expression (which often greatly exceeded that observable non-invasively during life). However, co-expression with enhanced GFP of myocilin or a juvenile glaucoma-associated mutant myocilin did not elevate IOP. These results demonstrate a safe, long-term single and dual gene expression in TM and establish an experimental system for testing candidate therapeutic transgenes for POAG. PMID- 17912237 TI - Hydrodynamic gene delivery: its principles and applications. AB - Efficient and safe methods for delivering genetic materials into cells must be developed before the clinical potential of gene therapy can be fully realized. Recently, hydrodynamic gene delivery using a rapid injection of a relatively large volume of DNA solution has opened up a new avenue for gene therapy studies in vivo. This method is superior to the existing delivery systems because of its simplicity, efficiency, and versatility. Wide success in applying hydrodynamic principles to delivery of DNA, RNA, proteins, and synthetic compounds, into the cells in various tissues of small animals, has inspired the recent attempts at establishing a hydrodynamic procedure for clinical use. In this review, we provide an overview of the theory and practice of hydrodynamic gene delivery so as to aid researchers for the use of this method in their pre-clinical and translational gene therapy studies. PMID- 17912238 TI - X-ray screening seems to reduce gastric cancer mortality by half in a community controlled trial in Costa Rica. AB - X-ray screening of gastric cancer is broadly used in Japan, although no controlled trial has proved its effectiveness. This study evaluates the impact of an X-ray screening demonstrative intervention to reduce gastric cancer mortality in a Costa Rican region. The evaluation follows a quasi-experimental, community controlled design, with measures before and after. About 7000 individuals participated by invitation in the two-wave screening programme. X-ray screening was followed by videoendoscopy and gastric biopsies. Treatment included resection with or without lymph node dissection. Comparisons with two control groups estimate that gastric cancer mortality was halved in the period from 2 to 7 years after the first screening visit. Validity of X-rays as used in this intervention had 88% sensitivity, 80% specificity, and 3% predictive value for individuals with two screening visits. Incidence in the screened group increased up to four times. Case survival was 85% in the intervention group after 5 years, compared to 12% among the controls before the intervention and 35% among the controls in the same region after the intervention. Although X-ray mass screening seems able to reduce stomach cancer mortality, its high cost may be an obstacle for scaling up this intervention in a non-rich country like Costa Rica. PMID- 17912239 TI - Mucin impedes cytotoxic effect of 5-FU against growth of human pancreatic cancer cells: overcoming cellular barriers for therapeutic gain. AB - Mucins are high molecular weight glycoproteins expressed on the apical surface of normal epithelial cells. In cancer disease mucins are overexpressed on the entire cellular surface. Overexpression of MUC1 mucin in pancreatic tumours has been correlated with poor patient survival. Current chemotherapeutic approaches such as 5-fluorouracil (5-FU) has produced limited clinical success. In this study we investigated the role of mucin in cytotoxic drug treatment to determine whether the extracellular domain of mucin impedes cytotoxic drug action of 5-FU. Human pancreatic cancer cells revealed high and relatively moderate MUC1 levels for Capan-1 and HPAF-II, respectively, compared to MUC1 negative control (U-87 MG glioblastoma) that showed relatively non-specific anti-MUC1 uptake. Benzyl-alpha GalNAc (O-glycosylation inhibitor) was used to reduce mucin on cell surfaces, and neuraminidase was used to hydrolyse sialic acid at the distal end of carbohydrate chains. Benzyl-alpha-GalNAc had no effect on cell morphology or proliferation at the concentrations employed. The inhibition of O-glycosylation resulted in significant 5-FU antiproliferative activity against Capan-1 and HPAF-II, but not against U-87 MG. However, the exposure of cells to neuraminidase failed to improve the cytotoxic action of 5-FU. Our experimental findings suggest that the overexpression of mucin produced by human pancreatic tumours might limit the effectiveness of chemotherapy. PMID- 17912240 TI - ZD6474 reverses multidrug resistance by directly inhibiting the function of P glycoprotein. AB - P-glycoprotein (P-gp) pumps multiple types of drugs out of the cell, using energy generated from ATP, and confers multidrug resistance (MDR) on cancer cells. ZD6474 is an orally active, selective inhibitor of the vascular endothelial growth factor receptor, epidermal growth factor receptor, and rearranged during transfection tyrosine kinases. This study was designed to examine whether ZD6474 reverses P-gp-mediated MDR in cancer cells. Here, we show that clinically achievable levels of ZD6474 reverse P-gp-mediated MDR of the P-gp-overexpressing cell lines derived from breast cancer, MCF-7/adriamycin (ADR), and human oral epidermoid carcinoma, KBV200 to ADR, docetaxel, and vinorelbine. This ability to reverse the P-gp-mediated resistance is comparable to that of another frequently used reversal agent known as verapamil. ZD6474 itself moderately inhibits the proliferation of both MCF-7 and MCF-7/ADR cells with almost equal activity, but its inhibitory effect is not altered by co-incubation with verapamil, suggesting that ZD6474 may not be a substrate of P-gp. In addition, ZD6474 increases the intracellular accumulation of the P-gp substrate, rhodamine-123, and ADR, by enhancing the uptake and/or decreasing the efflux of these compounds in resistant cells. Further studies show that ZD6474 stimulates ATPase activity in a dose dependent manner, which is required for the proper function of P-gp. In contrast, ZD6474 does not inhibit the expression level of P-gp. Our results suggest that ZD6474 is capable of reversing MDR in cancer cells by directly inhibiting the function of P-gp, a finding that may have clinical implications for ZD6474. PMID- 17912241 TI - Elevated serum matrix metalloproteinase 9 (MMP-9) concentration predicts the presence of colorectal neoplasia in symptomatic patients. AB - Early detection of polyps or colorectal carcinoma can reduce colorectal carcinoma associated deaths. Previous studies have demonstrated raised serum levels of matrix metalloproteinase 9 (sMMP-9) in a range of cancers. The aim of this study was to investigate the role of sMMP-9 levels in identifying colorectal neoplasia. Consenting patients donated a blood sample and were assessed by proforma-led history and physical examination. Samples were analysed for sMMP-9 concentration (enzyme-linked immuno-sorbant assay) and compared to final diagnoses. Logistic regression modelling determined independent factors associated with neoplasia. A total of 365 patients were recruited of whom 300 were analysed, including 46 normal controls. A total of 27 significant adenomas and 63 malignancies were identified. The median sMMP-9 concentration was 443 ng ml(-1) (IQR: 219-782; mean: 546). Patients with neoplasia had significantly elevated sMMP-9 levels (P<0.001). Logistic regression modelling identified elevated log(sMMP-9) as the most significant predictor of neoplasia (chi(2)=38.33, P<0.001). Other significant factors were age, sex, smoking history, abdominal pain and weight loss. The model accurately predicted neoplasia in 77.3% of cases. Sensitivity and specificity were 77.9 and 77.1%. sMMP-9 estimation can accurately stratify patient to low- or high-risk cohorts. Serum sampling is a potential means of avoiding unnecessary colonoscopy and reducing patient anxiety, iatrogenic morbidity and mortality, and cost. PMID- 17912242 TI - Where is VEGF in the body? A meta-analysis of VEGF distribution in cancer. AB - Vascular endothelial growth factor (VEGF) is a major target for the inhibition of tumour vascularisation and the treatment of human cancer. Many tumours produce large quantities of VEGF, and as a result, diagnosis and prognosis of cancer may be predicted by measuring changes in VEGF concentrations in blood. In blood, the VEGF may be located in the plasma, or in the blood-borne cells and formed elements, in particular, platelets and leukocytes. In this study, we collate the measurements of VEGF in platelets, leukocytes, plasma and serum for breast, prostate, colorectal and other cancers. In addition, we analysed the concentration of VEGF in tumour tissue itself, as well as for other tissues in the human body. Although the concentration of VEGF in tumours is high, the size of tumours is small compared to other tissues, in particular, skeletal muscle. Thus, the total quantity of VEGF in tumours and in blood is small compared to the quantity in muscles. This large reservoir of VEGF may have important implications for the treatment of cancer. PMID- 17912243 TI - Diabetes and endometrial cancer: effect modification by body weight, physical activity and hypertension. AB - Among 777 endometrial cancer cases and 1550 controls from Italy and Switzerland, odds ratio was 1.7 (95% confidence interval: 1.2-2.5) for diabetes, and 5.1 for obese diabetic women as compared with non-obese non-diabetic ones. Diabetes shows a supramultiplicative effect with body mass index, but not with physical activity or hypertension. PMID- 17912244 TI - C4.4A as a candidate marker in the diagnosis of colorectal cancer. AB - C4.4A is a member of the Ly-6 family with restricted expression in non transformed tissues. C4.4A expression in human cancer has rarely been evaluated. Thus, it became important to explore C4.4A protein expression in human tumour tissue to obtain an estimate on the frequency of expression and the correlation with tumour progression, the study focusing on colorectal cancer. The analysis of C4.4A in human tumour lines by western blot and immunoprecipitation using polyclonal rabbit antibodies that recognize different C4.4A epitopes revealed C4.4A oligomer and heavily glycosylated C4.4A isoform expression that, in some instances, inhibited antibody binding and interaction with the C4.4A ligand galectin-3. In addition, tumour cell lines released C4.4A by vesicle shedding and proteolytic cleavage. C4.4A was expressed in over 80% of primary colorectal cancer and liver metastasis with negligible expression in adjacent colonic mucosa, inflamed colonic tissue and liver. This compares well with EpCAM and CO 029 expression in over 90% of colorectal cancer. C4.4A expression was only observed in about 50% of pancreatic cancer and renal cell carcinoma. By de novo expression in colonic cancer tissue, we consider C4.4A as a candidate diagnostic marker in colorectal cancer, which possibly can be detected in body fluids. PMID- 17912245 TI - Cigarette smoking and breast cancer risk: a population-based study in Sweden. AB - In a Swedish population-based case-control study, smoking showed no convincing association with risk of postmenopausal breast cancer - regardless of timing or level of smoking exposure - either overall or among subgroups. PMID- 17912246 TI - Pemetrexed pharmacokinetics and pharmacodynamics in a phase I/II study of doublet chemotherapy with vinorelbine: implications for further optimisation of pemetrexed schedules. AB - The purpose of this study was to investigate the utility of plasma pharmacokinetic and pharmacodynamic measures including plasma deoxynucleosides, homocysteine and methylmalonic acid concentrations in understanding the time course and extent of the inhibition of thymidylate synthase (TS) by pemetrexed in the context of a phase I/II combination study with vinorelbine. Eighteen patients received supplementation with folic acid and Vitamin B(12) 1 week before beginning treatment with pemetrexed and vinorelbine administered in a dose escalating manner on a 21-day cycle. Heparinised blood samples were collected from consenting patients in the first cycle for pharmacokinetic analyses and in the first two cycles for determination of plasma thymidine, deoxyuridine, homocysteine and methylmalonic acid concentrations. These values were correlated with response and toxicity. Plasma deoxyuridine was used as a measure of TS inhibition, and concentrations of deoxyuridine were significantly elevated relative to baseline on days 1 (P<0.01), 2 (P<0.001) and 3 (P<0.05) after treatment at all pemetrexed dose levels (400-700 mg m(-2)). The magnitude of deoxyuridine elevation correlated with pemetrexed area under the plasma concentration-time curve (AUC) (r(2)=0.23, P<0.05). However, deoxyuridine concentrations returned to baseline between 8 and 15 days after treatment with pemetrexed, suggesting that inhibition of TS was not durable. Pemetrexed AUC correlated with the percentage decline (relative to baseline) in both platelets (r(2)=0.58, P<0.001) and leucocytes (r(2)=0.26, P<0.05) at day 8. Baseline homocysteine was also significantly correlated with these measures of haematological toxicity (r(2)=0.37, P<0.01 and r(2)=0.39, P<0.01, respectively). In addition, there was a significant reduction of plasma homocysteine on days 8 (P<0.005) and 15 (P<0.05) in cycle 1 compared to baseline values. The results suggest that the TS inhibitory effects of pemetrexed are short-lived and make the case for a more frequent schedule of administration such as every 2 weeks. The lack of protracted TS inhibition may be due to concomitant vitamin administration, and this may be the mechanism by which vitamins prevent life threatening toxicity from pemetrexed. Baseline homocysteine concentration remains a predictive marker for haematological toxicity even following folate supplementation. PMID- 17912247 TI - Migration-promoting role of VEGF-C and VEGF-C binding receptors in human breast cancer cells. AB - Vascular endothelial growth factor C (VEGF-C) is a lymphangiogenic factor over expressed in highly metastatic, cyclooxygenase (COX)-2 expressing breast cancer cells. We tested the hypothesis that tumour-derived VEGF-C may play an autocrine role in metastasis by promoting cellular motility through one or more VEGF-C binding receptors VEGFR-2, VEGFR-3, neuropilin (NRP)-1, NRP-2, and integrin alpha9beta1. We investigated the expression of these receptors in several breast cancer cell lines (MDA-MB-231, Hs578T, SK-BR-3, T-47D, and MCF7) and their possible requirement in migration of two VEGF-C-secreting, highly metastatic lines MDA-MB-231 and Hs578T. While cell lines varied significantly in their expression of above VEGF-C receptors, migratory activity of MDA-MB-231 and Hs578T cells was linked to one or more of these receptors. Depletion of endogenous VEGF C by treatments with a neutralising antibody, VEGF-C siRNA or inhibitors of Src, EGFR/Her2/neu and p38 MAP kinases which inhibited VEGF-C production, inhibited cellular migration, indicating the requirement of VEGF-C for migratory function. Migration was differentially attenuated by blocking or downregulation of different VEGF-C receptors, for example treatment with a VEGFR-2 tyrosine kinase inhibitor, NRP-1 and NRP-2 siRNA or alpha9beta1 integrin antibody, indicating the participation of one or more of the receptors in cell motility. This novel role of tumour-derived VEGF-C indicates that breast cancer metastasis can be promoted by coordinated stimulation of lymphangiogenesis and enhanced migratory activity of breast cancer cells. PMID- 17912248 TI - The DISC locus in psychiatric illness. AB - The DISC locus is located at the breakpoint of a balanced t(1;11) chromosomal translocation in a large and unique Scottish family. This translocation segregates in a highly statistically significant manner with a broad diagnosis of psychiatric illness, including schizophrenia, bipolar disorder and major depression, as well as with a narrow diagnosis of schizophrenia alone. Two novel genes were identified at this locus and due to the high prevalence of schizophrenia in this family, they were named Disrupted-in-Schizophrenia-1 (DISC1) and Disrupted-in-Schizophrenia-2 (DISC2). DISC1 encodes a novel multifunctional scaffold protein, whereas DISC2 is a putative noncoding RNA gene antisense to DISC1. A number of independent genetic linkage and association studies in diverse populations support the original linkage findings in the Scottish family and genetic evidence now implicates the DISC locus in susceptibility to schizophrenia, schizoaffective disorder, bipolar disorder and major depression as well as various cognitive traits. Despite this, with the exception of the t(1;11) translocation, robust evidence for a functional variant(s) is still lacking and genetic heterogeneity is likely. Of the two genes identified at this locus, DISC1 has been prioritized as the most probable candidate susceptibility gene for psychiatric illness, as its protein sequence is directly disrupted by the translocation. Much research has been undertaken in recent years to elucidate the biological functions of the DISC1 protein and to further our understanding of how it contributes to the pathogenesis of schizophrenia. These data are the main subject of this review; however, the potential involvement of DISC2 in the pathogenesis of psychiatric illness is also discussed. A detailed picture of DISC1 function is now emerging, which encompasses roles in neurodevelopment, cytoskeletal function and cAMP signalling, and several DISC1 interactors have also been defined as independent genetic susceptibility factors for psychiatric illness. DISC1 is a hub protein in a multidimensional risk pathway for major mental illness, and studies of this pathway are opening up opportunities for a better understanding of causality and possible mechanisms of intervention. PMID- 17912249 TI - The amyloidogenic potential and behavioral correlates of stress. AB - Observations of elevated basal cortisol levels in Alzheimer's disease (AD) patients prompted the hypothesis that stress and glucocorticoids (GC) may contribute to the development and/or maintenance of AD. Consistent with that hypothesis, we show that stress and GC provoke misprocessing of amyloid precursor peptide in the rat hippocampus and prefrontal cortex, resulting in increased levels of the peptide C-terminal fragment 99 (C99), whose further proteolytic cleavage results in the generation of amyloid-beta (Abeta). We also show that exogenous Abeta can reproduce the effects of stress and GC on C99 production and that a history of stress strikingly potentiates the C99-inducing effects of Abeta and GC. Previous work has indicated a role for Abeta in disruption of synaptic function and cognitive behaviors, and AD patients reportedly show signs of heightened anxiety. Here, behavioral analysis revealed that like stress and GC, Abeta administration causes spatial memory deficits that are exacerbated by stress and GC; additionally, Abeta, stress and GC induced a state of hyperanxiety. Given that the intrinsic properties of C99 and Abeta include neuroendangerment and behavioral impairment, our findings suggest a causal role for stress and GC in the etiopathogenesis of AD, and demonstrate that stressful life events and GC therapy can have a cumulative impact on the course of AD development and progression. PMID- 17912250 TI - Behavioral and pharmacological validation of the gerbil forced-swim test: effects of neurokinin-1 receptor antagonists. AB - Several studies have suggested that neurokinin-1 (NK1) receptor antagonists may have therapeutic potential as novel antidepressant drugs. To test these compounds preclinically, gerbils have become one of the preferred species in that they demonstrate close NK1 receptor homology with humans and bind NK1 antagonists with higher affinity than rats and mice. The intent of the present study was to determine whether the forced-swim test (FST), one of the most commonly used animal tests of antidepressant-like activity, could be adapted for use with the gerbil. Critical factors in the establishment of this assay included swim tank diameter, weight, and sex of the animals tested. Pharmacological validation of the FST using standard antidepressant compounds (eg fluoxetine, paroxetine, desipramine) resulted in decreased immobility time during the test, indicative of an antidepressant-like effect. Similar to results reported for the rat and mouse FST, the antipsychotic drug haloperidol increased immobility, whereas the psychostimulant, amphetamine decreased immobility, and anxiolytic drugs (eg buspirone) had no effect. Investigation into the locomotor effects of all compounds tested was consistent with previous reports in other species, with the exception of paroxetine, which produced hyperactivity at therapeutically effective doses in gerbils. In addition to standard antidepressants, NK1 antagonists (L-733060, MK-869, and CP-122721) all reduced immobility in the gerbil FST without affecting locomotor activity. Overall, these results suggest that the gerbil is an ideal species for use in the FST, and that this paradigm may have predictive validity for identifying novel antidepressant compounds. PMID- 17912251 TI - Cellular plasticity cascades in the pathophysiology and treatment of bipolar disorder. AB - Bipolar disorder (BPD) is characterized by recurrent episodes of disturbed affect including mania and depression as well as changes in psychovegetative function, cognitive performance, and general health. A growing body of data suggests that BPD arises from abnormalities in synaptic and neuronal plasticity cascades, leading to aberrant information processing in critical synapses and circuits. Thus, these illnesses can best be conceptualized as genetically influenced disorders of synapses and circuits rather than simply as deficits or excesses in individual neurotransmitters. In addition, commonly used mood-stabilizing drugs that are effective in treating BPD have been shown to target intracellular signaling pathways that control synaptic plasticity and cellular resilience. In this article we draw on clinical, preclinical, neuroimaging, and post-mortem data to discuss the neurobiology of BPD within a conceptual framework while highlighting the role of neuroplasticity in the pathophysiology and treatment of this disorder. PMID- 17912253 TI - Allele-specific regulation of primary cilia function by the von Hippel-Lindau tumor suppressor. AB - Patients with von Hippel-Lindau (VHL) disease often develop VHL-/- kidney cysts, which possibly progress into clear-cell renal carcinomas (ccRCCs). Recent data link the VHL gene product to formation of the primary cilium, an organelle that extends apically into the renal lumen. Exactly how VHL induces ciliogenesis or function is unknown. Here, we demonstrate that ciliary assembly and mechanotransduction is rapidly restored in VHL-/- ccRCC cells upon ectopic reconstitution of wild-type - but not variant alleles of - VHL. These data support and expand recent studies implicating a role for VHL in the initiation of ciliogenesis. Furthermore, reduction of cellular levels of VHL in this cell system was associated with fewer ciliated cells, suggesting a role for VHL in ciliary maintenance. PMID- 17912254 TI - AH receptor agonist activity in human blood measured with a cell-based bioassay: evidence for naturally occurring AH receptor ligands in vivo. AB - In the present study, an aryl hydrocarbon receptor (AHR)-driven reporter gene bioassay was used to measure the activity, measured as an induction equivalent (IEQ) as compared to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), or IEQ concentration in human blood samples from 10 volunteers under different dietary regimens. Blood concentrations of polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs) and polychlorinated biphenyls (PCBs), as determined by analytical chemistry (HR-GC/MS), and expressed as toxic equivalents (TEQs) with the use of TCDD equivalency factors (TEFs), were within a range that has been reported in the general US population, ranging from 0.022 to 0.119 ppt (whole blood basis). However, the human blood IEQ measured directly via bioassay ranged from 13.4 to 218 ppt (whole blood basis). These order of magnitude greater IEQs compared to the TEQs for dioxins, furans, and certain PCBs suggests that human blood contains a relatively high level of AHR agonists able to activate the CYP1A1 dioxin response element (DRE)-linked reporter gene bioassay and that this AHR activity is not accounted for by PCDDs/Fs and dioxin-like PCBs based on standard HR-GC/MS and TEF analysis. When study participants switched from a "baseline" to a high vegetable diet, increases in bioassay IEQ were observed that were statistically significant (P<0.05). In addition, IEQ activity was elevated above levels observed following dietary intervention in two subjects given indole-3-carbinol (I3C) supplements. We conclude that a substantial portion of the IEQ activity occurred as a result of the increased intake of natural AHR agonists (NAHRAs) present in many fruits, vegetables. and herbs. Our findings also suggest that dietary NAHRAs constitute a substantial daily dietary intake of AHR-active compounds, and these NAHRAs could influence AHR status in humans and play a role in a basal level of AHR activation. PMID- 17912255 TI - New partners for BRCA1. PMID- 17912252 TI - Divergent plasticity of prefrontal cortex networks. AB - The 'executive' regions of the prefrontal cortex (PFC) such as the dorsolateral PFC (dlPFC) and its rodent equivalent medial PFC (mPFC) are thought to respond in concert with the 'limbic' regions of the PFC such as the orbitofrontal (OFC) cortex to orchestrate behavior that is consistent with context and expected outcome. Both groups of regions have been implicated in behavioral abnormalities associated with addiction and psychiatric disorders, in particular, schizophrenia and mood disorders. Theories about the pathophysiology of these disorders, however, incorporate abnormalities in discrete PFC regions independently of each other or assume they are one and the same and, thus, bunch them under umbrella of 'PFC dysfunction.' Emerging data from animal studies suggest that mPFC and OFC neurons display opposing patterns of plasticity during associative learning and in response to repeated exposure to psychostimulants. These data corroborate clinical studies reporting different patterns of activation in OFC versus dlPFC in individuals with schizophrenia or addictive disorders. These suggest that concomitant but divergent engagement of discrete PFC regions is critical for learning stimulus-outcome associations, and the execution of goal-directed behavior that is based on these associations. An atypical interplay between these regions may lead to abnormally high or low salience assigned to stimuli, resulting in symptoms that are fundamental to many psychiatric and addictive disorders, including attentional deficits, improper affective response to stimuli, and inflexible or impulsive behavior. PMID- 17912256 TI - How much can SNAREs flex their muscles? PMID- 17912257 TI - Prion strains under the magnifying glass. PMID- 17912258 TI - Multienzyme assembly of a p53 transcription complex. PMID- 17912259 TI - Sensing of DNA damage by XPC/Rad4: one protein for many lesions. PMID- 17912263 TI - Finishing mitosis, one step at a time. AB - The final stages of mitosis begin in anaphase, when the mitotic spindle segregates the duplicated chromosomes. Mitotic exit is then completed by disassembly of the spindle and packaging of chromosomes into daughter nuclei. The successful completion of mitosis requires that these events occur in a strict order. Two main mechanisms govern progression through late mitosis: dephosphorylation of cyclin-dependent kinase (Cdk) substrates and destruction of the substrates of the anaphase-promoting complex (APC). Here, we discuss the hypothesis that the order of late mitotic events depends, at least in part, on the order in which different Cdk and APC substrates are dephosphorylated or destroyed, respectively. PMID- 17912264 TI - Vacuolar ATPases: rotary proton pumps in physiology and pathophysiology. AB - The acidity of intracellular compartments and the extracellular environment is crucial to various cellular processes, including membrane trafficking, protein degradation, bone resorption and sperm maturation. At the heart of regulating acidity are the vacuolar (V-)ATPases--large, multisubunit complexes that function as ATP-driven proton pumps. Their activity is controlled by regulating the assembly of the V-ATPase complex or by the dynamic regulation of V-ATPase expression on membrane surfaces. The V-ATPases have been implicated in a number of diseases and, coupled with their complex isoform composition, represent attractive and potentially highly specific drug targets. PMID- 17912265 TI - Neuroblastoma, childhood cancer survivorship, and reducing the consequences of cure. PMID- 17912266 TI - Mexican anthropometry percentiles and the LMS method. PMID- 17912267 TI - A prospective analysis of dietary energy density at age 5 and 7 years and fatness at 9 years among UK children. AB - OBJECTIVE: To analyse whether high dietary energy density (DED) is associated with increased fat mass and risk of excess adiposity in free-living children. DESIGN: Longitudinal, observational cohort study. SUBJECTS: Six hundred and eighty-two healthy children from the Avon Longitudinal Study of Parents and Children. MEASUREMENTS: Diet was assessed at age 5 and 7 years using 3-day diet diaries, and DED (kJ g(-1)) was calculated excluding drinks. Fat mass was estimated at age 9 years using Dual-Energy X-ray Absorptiometry. To adjust for body size, fat mass index (FMI) was calculated by dividing fat mass (kg) by height (m(5.8)). Excess adiposity was defined as the top quintile of logFMI. RESULTS: Mean DED at age 5 years was higher among children with excess adiposity at age 9 years compared to the remaining sample (8.8+/-0.16 vs 8.5+/-0.07 kJ g( 1)), but there was no evidence of an association with excess adiposity at age 9 years (odds ratio (OR)=1.14, 95% confidence interval (CI) 0.90-1.44) after controlling for potential confounders. Mean DED at age 7 years was higher among children with excess adiposity compared to the remaining sample (9.1+/-0.12 vs 8.8+/-0.06 kJ g(-1)) and a 1 kJ g(-1) rise in DED increased the odds of excess adiposity at 9 years by 36% (OR=1.36, 95% CI 1.09-1.69) after controlling for potential confounders. CONCLUSION: Higher DED at age 7 years, but not age 5 years, is a risk factor for excess adiposity at age 9 years, perhaps reflecting deterioration in the ability to compensate for extra calories in an energy-dense diet. DED tracks strongly from age 5 to 7 years suggesting intervention to alter dietary habits need to commence at younger ages to prevent the formation of preferences for energy dense foods. PMID- 17912268 TI - Search for genetic variants of the SYNTAXIN 1A (STX1A) gene: the -352 A>T variant in the STX1A promoter associates with impaired glucose metabolism in an Italian obese population. AB - OBJECTIVE: To test if sequence variations of the SYNTAXIN 1A (STX1A) gene contribute to the susceptibility to type 2 diabetes in a cohort of overweight/obese subjects. METHODS: A total of 717 overweight/obese individuals underwent oral glucose tolerance test and were stratified in four groups according to fasting and 2 h glucose levels (NGT, IGT, CGI, T2DM), representing the natural history of diabetes from normal glucose tolerance to overt disease. These subjects were analysed by a two-step genetic study. Functional analysis was performed by electrophoretic mobility shift assay (EMSA) and by supershift with CCAAT/enhancer-binding protein (C/EBP)beta antibody. RESULTS: Among the several sequence variations detected in the STX1A gene, the T allele of the -352 A>T single nucleotide polymorphism in the promoter was found in a lower frequency in the subset of individuals with greater impairment of insulin secretion (CGI). To confirm that a lower frequency of the T allele was associated with this condition, we genotyped a second group of 202 overweight/obese individuals with type 2 diabetes, and the frequency of the T allele was reduced in this group also (P<0.01). Logistic regression confirmed a protective odds ratio (0.49, P<0.01) for the T allele. The EMSA showed that the PRM -352 A allele binds transcription factors with lower affinity compared to the T allele, and incubation with C/EBPbeta antibody 'supershifted' the complex, indicating that C/EBPbeta had a different binding with the PRM -352T allele. CONCLUSION: A lower frequency of the PRM -352T allele of the STX1A gene was observed in overweight/obese subjects with impaired glucose regulation, particularly among individuals with combined glucose intolerance and overt diabetes. Both these groups have a greater defect in beta cell function compared to normal and glucose intolerant subjects, and this association together with the functional study suggests a possible role of the PRM -352 A>T variant in insulin secretion. PMID- 17912269 TI - Peanut digestion and energy balance. AB - OBJECTIVE: To explore the effects of peanut consumption on fecal energy excretion with a balanced, non-vegetarian diet. DESIGN: Four arm parallel group design (that is, whole peanut (P), peanut butter (PB), peanut oil (PO) or peanut flour (PF) consumption) with one crossover (control and intervention). SUBJECTS: In total 63 healthy men and women from Ghana, Brazil and USA (N=15-16 per group) with an average body mass index of 21.8 kg m(-2). MEASUREMENTS: Percent fat of fecal wet weight daily energy excretion during the control and the treatment periods. RESULTS: Compared to control, the percentage of fat in the feces increased significantly for the P group (5.22+/-0.29%) relative to the other three groups ((PO=3.07+/-0.36%, PB=3.11+/-0.31% (P=0.001), and PF=3.75+/-0.40% (P=0.019)). The same findings held for kJ g(-1) of feces excreted. During the P supplementation period, the energy excretion was 21.4+/-1.0 kJ g(-1) versus 18.7+/-1.0 kJ g(-1) for PO (P=0.034), 18.8+/-0.7 kJ g(-1) for PB (P=0.042) and 18.5+/-0.8 kJ g(-1) for PF (P=0.028). CONCLUSION: Fecal fat and energy loss is greater with consumption of whole peanuts compared to peanut butter, oil or flour. This may contribute to the less than predicted change of body weight observed with peanut consumption. There were no cultural differences. PMID- 17912270 TI - Total cerebral blood flow, white matter lesions and brain atrophy: the SMART-MR study. AB - We investigated whether total cerebral blood flow (CBF) was associated with brain atrophy, and whether this relation was modified by white matter lesions (WML). Within the Second Manifestations of ARTerial disease-magnetic resonance (SMART MR) study, a prospective cohort study among patients with arterial disease, cross sectional analyses were performed in 828 patients (mean age 58+/-10 years, 81% male) with quantitative flow, atrophy, and WML measurements on magnetic resonance imaging (MRI). Total CBF was measured with MR angiography and was expressed per 100 mL brain volume. Total brain volume and ventricular volume were divided by intracranial volume to obtain brain parenchymal fraction (BPF) and ventricular fraction (VF). Lower BPF indicates more global brain atrophy, whereas higher VF indicates more subcortical brain atrophy. Mean CBF was 52.0+/-10.2 mL/min per 100 mL, mean BPF was 79.2+/-2.9%, and mean VF was 2.03+/-0.96%. Linear regression analyses showed that lower CBF was associated with more subcortical brain atrophy, after adjusting for age, sex, vascular risk factors, intima-media thickness, and lacunar infarcts, but only in patients with moderate to severe WML (upper quartile of WML): Change in VF per s.d. decrease in CBF 0.18%, 95% CI: 0.02 to 0.34%. Our findings suggest that cerebral hypoperfusion in the presence of WML may be associated with subcortical brain atrophy. PMID- 17912271 TI - A concerted role of Na+ -K+ -Cl- cotransporter and Na+/Ca2+ exchanger in ischemic damage. AB - Na+-K+-Cl(-) cotransporter isoform 1 (NKCC1) and Na+/Ca2+ exchanger isoform 1 (NCX1) were expressed in cortical neurons. Three hours of oxygen and glucose deprivation (OGD) significantly increased expression of full-length NCX1 protein ( approximately 116 kDa), which remained elevated during 1 to 21 h reoxygenation (REOX) and was accompanied with concurrent cleavage of NCX1. Na+/Ca2+ exchanger isoform 1 heterozygous (NCX1+/-) neurons with approximately 50% less of NCX1 protein exhibited approximately 64% reduction in NCX-mediated Ca2+ influx. Expression of NCX1 and NKCC1 proteins was reduced in double heterozygous (NCX1+/ /NKCC1+/-) neurons. NCX-mediated Ca2+ influx was nearly abolished in these neurons. Three-hour OGD and 21-h REOX caused approximately 80% mortality rate in NCX1+/+ neurons and in NCX1+/- neurons. In contrast, NKCC1+/- neurons exhibited approximately 45% less cell death. The lowest mortality rate was found in NCX1+/ /NKCC1+/- neurons ( approximately 65% less neuronal death). The increased tolerance to ischemic damage was also observed in NCX1+/-/NKCC1+/- brains after transient cerebral ischemia. NCX1+/-/NKCC1+/- mice had a significantly reduced infarct volume at 24 and 72 h reperfusion. In conclusion, these data suggest that NKCC1 in conjunction with NCX1 plays a role in reperfusion-induced brain injury after ischemia. PMID- 17912272 TI - The mechanisms of acute ischemic injury in the cell processes of developing white matter astrocytes. AB - Astrocytes are fundamentally important to the maintenance and proper functioning of the central nervous system. During the period of development when myelination is occurring, white matter astrocytes are particularly sensitive to ischemic injury and their failure to regulate glutamate during ischemic conditions may be an important factor in excitotoxic injury. Here, we have identified key mechanisms of injury that operate on the processes of immature white matter astrocytes during oxygen-glucose deprivation (OGD) using GFAP-GFP mice. Oxygen glucose deprivation produced a parallel loss of astrocyte processes and somata, assessed by both the retention of GFP fluorescence within these structures and by quantitative electron microscopy. Oxygen-glucose deprivation-induced process loss was Ca(2+) independent and had two distinct mechanisms. Substituting either extracellular Na(+) or Cl(-), or perfusion with the Na-K-Cl co-transport blocker bumetanide, provided protection up to 40 mins of OGD but not beyond that point. HCO(3)(-) substitution or perfusion with 4,4'-diisothiocyanostilbene-2,2' disulphonic acid provided complete protection of the processes up to 60 mins of OGD. Zero-Na(+)/zero-K(+) conditions provided complete protection from OGD induced injury of processes and somata at all time points. We conclude that acute ischemic-type injury of immature astrocytes follows a cytotoxic ion influx mediated in part by Na-K-Cl co-transport and in part by Na(+)- and K(+)-dependent HCO(3)(-) transport, a mechanism that is common to both cell processes and somata. This work provides a basis on which preventative strategies may be developed to protect white matter astrocytes from ischemic injury in susceptible individuals. PMID- 17912273 TI - [Inequalities in public health]. PMID- 17912274 TI - [Prevalence of Staphylococcus aureus nasal colonization in injection drug users]. AB - A prevalence study was conducted to evaluate Staphylococcus aureus nasal colonization in two groups of injection drug users recruited at two drug addiction treatment centres in Rome and Frosinone (Italy). Prevalence of Staphylococcus aureus nasal colonization in the 181 subjects included in the study was 8.83%. Significantly increased carriage rates were found among drug users in Frosinone with respect to those in Rome. No other significant determinants of colonization were identified through multiple logistic regression analysis. Only 2 of 16 Staphylococcus aureus strains (12.5%) were found to be methicillin resistant. Genomic DNA restriction pattern analysis, through SmaI digestion and pulsed field gel electrophoresis, demonstrated wide genetic heterogeneity among the isolates. Although the great majority of PFGE patterns were found in single isolates, in some cases common patterns were identified in isolates obtained from different drug users, clearly indicating the possibility of interhuman transmission of Staphylococcus aureus in this population. PMID- 17912275 TI - [Clinical risk prevention by coordinator nurses, knowledge and organizational tools]. AB - Coordinator nurses have a pivotal role to play in clinical risk prevention. They can and must influence the design of work processes to promote a culture of safety through the use of checklists, protocols, decision support systems, and by improving access to information, standardising tasks, and reducing the number of "hand-overs" and tasks performed relying only on one's memory. In order to evaluate knowledge and use of organisational tools for clinical risk prevention by coordinator nurses, an exploratory questionnaire survey was performed at a university teaching hospital in central Italy. Approximately 100 coordinators were asked to participate in the study. The aims of the survey were to assess the extent to which certain clinical governance tools are being used in individual hospital departments, evaluate knowledge of organisational processes that lead to errors and of measures, pertaining to nursing, that are considered to be effective in reducing adverse events. Wide differences were found among departments in the use of risk management strategies. Results also show that many nurse coordinators are not clearly aware of their organisational and professional responsibilities in clinical risk management. Though many professionals are well qualified and knowledgeable, only few are prepared educationally to manage clinical risk effectively when delivering patient care. For this reason, leadership should be encouraged and sustained, through educational courses and the promotion of a shift towards a culture of safety substantiated by scientific evidence. PMID- 17912276 TI - [Food hygiene training of members of corporate public catering committees]. AB - A food hygiene training course was offered to 25 members of the public catering committees of seven corporate restaurants. In order to evaluate the effectiveness of the course and identify critical topics, participants were asked to complete a questionnaire before and after completing the training course. Results are presented in this article and underscore the importance of training members of public catering committees in addition to food handlers. PMID- 17912277 TI - Which child health and non child health indicators are appropriate for programming health services at a district level? AB - The aim of this study was to identify which indicators measuring social and health status of children are most suitable for planning district health services in European countries. A search was performed for documentation regarding child health indicators currently used in Italy, the European community, in the United Kingdom and Canada. Nine indicators were selected and for each indicator, a reference value was identified by consulting data bases and reports, information regarding usefulness of the indicator is specified, and intervention strategies for improving quality of health care in the specific area are described. PMID- 17912278 TI - Techniques for the recovery of enteric viruses from artificially contaminated marine sediments. AB - Viruses are an important component of aquatic microbial communities and marine sediments may represent an optimal means for their survival. The aim of this study was to evaluate different methods for virus recovery from marine sediments. Three methods were used for virus recovery from artificially contaminated sediments: elution and centrifugation technique, sonication technique, and mechanical disgregation followed by elution and centrifuge technique. The sonication technique obtained the highest virus recovery percentages (94,25%). Eluent 2 provided more efficient recovery of enteric viruses than eluent 1 presumably due to the presence, in eluent 2, of NANO3, a chaotropic agent that enhances the solubilization of hydrophobic compounds in water. Finally, the authors confirm the importance of searching for viruses in sediments, which protect them from inactivation by biological, chemical and physical factors and allow them to survive longer than in the overlaying water column. PMID- 17912279 TI - [Guidelines for public health interventions]. PMID- 17912280 TI - [Metropolitan areas and human health]. PMID- 17912281 TI - [Screening and vaccination: two synergic tools for the prevention of cervical cancer]. AB - Cervical cancer is the second common cancer in women, caused by HPV infection in 99,7% of cases. Screening programs have strongly reduced the mortality. Anti-HPV vaccination against types 16 and 18, responsible for 70% of cases, could contribute to reduce the burden of disease. In Italy the mass immunisation strategy, planned for twelve years old girls, must be supported by correct information of both population and health care workers. PMID- 17912282 TI - Enhanced effect of local fields in subwavelength metallic series nanocavities from surface plasmon polaritons. AB - The enhancement and confinement characteristics of the local field in the two dimensional (2D) subwavelength-size series cavities structure are investigated numerically by using the boundary integral method. The series cavities are built of two pieces of finite silver thin slabs with subwavelength corrugations on their inner boundaries, set in a face-to-face arrangement with a separating space, and the central part is a narrow channel (NC). We calculate the average amplitude of the local field in the NC as a function of the wavelength for exploring the influence of the structural parameters and demonstrate the amplitude distribution of the magnetic field in the structure and the cross section distributions of the local field in the NC region along both the longitudinal axis direction and the transverse directions. The simulations show that the local field in the NC has significant enhancement, up to 2 orders of magnitude, of the incident light field, and the local light field is confined to a small region less than one fifth of the resonant wavelength in the longitudinal direction and one twentieth of the resonant wavelength in the lateral direction. Replacing the metallic material of the cavity walls with the semiconductor germanium leads to the complete disappearance of the enhancement of the local field. It is clearly shown that surface plasmon polaritons on the metal play a critical role for this enhancement phenomenon. The influences of various geometric parameters on the resonant wavelength and the peak value of the average amplitude of the local field are extensively investigated. PMID- 17912283 TI - Generalized scattering-matrix method for the analysis of two-dimensional photonic bandgap devices. AB - Development of accurate numerical methods for the analysis of photonic-bandgap based devices is a relevant issue in optimizing existing devices and/or developing new design solutions. Within this framework, we present an innovative and general approach for the evaluation of the electromagnetic behavior of two dimensional finite-extent photonic crystals made of a finite set of parallel rods. The proposed approach is a generalization of the scattering-matrix method introduced by Maystre and co-workers and of its improved version proposed by the present authors, which exploits a suitable aggregation into "macrocells" to achieve a reduction of the number of unknowns. As a matter of fact, both of these approaches can be exploited only in those cases in which particular modal expansions for the fields hold true. In order to overcome this limitation, we propose a suitable exploitation of the method of auxiliary sources to provide a general and reliable method for the numerical computation of the scattering matrix of an object of arbitrary shape. By taking advantage of this, we can then generalize our improved scattering matrix method to further increase its computational effectiveness. A numerical analysis of some square-lattice configurations is reported to confirm the accuracy of the proposed method and the remarkable computational benefit. PMID- 17912284 TI - Observation of Anderson localization of light in three dimensions. AB - Using time-resolved transmission measurements, we have found indications of Anderson localization of light in bulk three-dimensional systems. The observed deviation from classical diffusion is in good accord with theoretical predictions of localization and cannot be explained by absorption or experimental artifacts such as stratification, fluorescence, or background illumination. Moreover, we show that in our samples the control parameter is given by the mean free path times the wavenumber as required by the Ioffe-Regel criterion. This is in contrast to quasi-one-dimensional systems that were studied with microwaves. There, the control parameter is related to the number of modes inside a waveguide, and deviations from classical diffusion are possible due to a small number of modes. PMID- 17912285 TI - Omnidirectional and multiple-channeled high-quality filters of photonic heterostructures containing single-negative materials. AB - The design and application of omnidirectional and multiple-channeled filters with high-quality (high-Q) factors applied by using photonic heterostructures containing single-negative materials is demonstrated. Through adjusting the period number of the heterostructures, the number of the resonance modes can be controlled, and the resonance modes are then insensitive to the incident angle. With perfect transmission, controllable mode, and omnidirectional channel, this structure opens a promising way to fabricate omnidirectional and multiple channeled filters for future dense wavelength division multiplexing applications. The effect of the losses of single-negative materials on the filters is also considered. PMID- 17912286 TI - Observation of singularities in multiply scattered microwave fields. AB - Speckle patterns of arbitrary resolution are obtained by applying the sampling theorem to measurements of two orthogonal components of the microwave field transmitted through multiply scattering samples. Core structures of phase singularities, phase critical points, and polarization singularities are explored. We find that equiphase lines connect phase singularities with opposite topological signs except for the bifurcation lines, which run through a phase saddle point, in agreement with predictions by Freund [Phys. Rev. E25, 2348 (1995)]. We observe hyperbolic equiphase lines near phase saddle points and elliptical equiphase lines around phase extrema. Polarization singularities of the vector field with the three morphologies predicted are observed. PMID- 17912287 TI - Metamaterials with gradient negative index of refraction. AB - We propose a new metamaterial with a gradient negative index of refraction, which can focus a collimated beam of light coming from a distant object. A slab of the negative refractive index metamaterial has a focal length that can be tuned by changing the gradient of the negative refractive index. A thin metal film pierced with holes of appropriate size or spacing between them can be used as a metamaterial with the gradient negative index of refraction. We use finite difference time-domain calculations to show the focusing of a plane electromagnetic wave passing through a system of equidistantly spaced holes in a metal slab with decreasing diameters toward the edges of the slab. PMID- 17912288 TI - Image inversion and magnification by negative index prisms. AB - A Dove prism and an anamorphic prism pair are investigated in negative index imaging systems. An equilateral triangular prism with refractive index of -1 operates as a negative index Dove prism that inverts as well as images the incident field. A negative index anamorphic prism pair acts as a negative index imaging system with magnification. The relationship between achievable magnification and aberrations is discussed. Both prism systems can be implemented by using negative index photonic crystals, and their performance is demonstrated numerically by the finite-difference time-domain method. These negative index prism structures enhance the functionalities of negative index flat lenses and broaden the applications of negative index materials. PMID- 17912289 TI - Semiclassical theory of the hyperlens. AB - We study ray dynamics inside the hyperlens, a device capable of sub-diffraction limited far-field imaging. An analytical result for the ray trajectories inside the hyperlens is obtained using Hamiltonian optics, which offers an alternative description of the device. It is also found that the ray trajectories can exhibit a unique spiraling nature inside the device. Numerical simulations of plane wave and Gaussian beam scattering from the hyperlens confirm the semiclassical description. PMID- 17912290 TI - A periodic structure mimics a metamaterial. AB - We show that properties attributed to planar metamaterials that support resonances due to an appropriately shaped unit cell can also be identified in a medium that exhibits a resonance evoked by its period only. By choosing a subwavelength period, the effective material parameters of such a medium can be retrieved from the complex reflected and transmitted amplitude. The parameters exhibit Lorentzian line shapes in the spectral vicinity of the resonances associated with the period. If this material is stacked to form a three dimensional medium, a stop gap is observed in transmission in the frequency range where the real part of one effective material parameter becomes negative. The resonance at the origin of the response is related to the excitation of a higher order Bloch mode. Because their negligible excitation is a prerequisite for deriving effective material parameters, the analyzed structure mimics only the response of a metamaterial but cannot be regarded as a metamaterial. PMID- 17912291 TI - Channel drop filter in two-dimensional triangular lattice photonic crystals. AB - Based on two-dimensional photonic crystals with a triangular lattice, a channel drop filter with a wavelength-selective reflection microcavity is designed. In the structure, two microcavities are used. One is used for a resonant tunneling based channel drop operation. The other is used to realize wavelength-selective reflection feedback in the bus waveguide. The phase term, which is derived by means of coupled-mode theory to achieve close to 100% drop efficiency, is satisfied by modifying the sizes of the border air holes next to the bus waveguide section between the two cavities. Using the finite-difference time domain method, the simulation results show complete power transfer between the bus and drop waveguides via the system. PMID- 17912294 TI - Color modeling of stratified pictorial layers using the radiative transfer equation solved by the auxiliary function method. AB - The diffuse reflectance spectra and the trichromatic coordinates of diffusing stratified paints are modeled. Each layer contains its own pigments, and their optical properties are first determined from experiments. The radiative transfer equation is then solved by the auxiliary function method for modeling the total light scattered by the stratified systems. The results are in good agreement with experimental spectra and validate the modeling. The calculations are then applied on the same stratified systems to study the influence of the observation angle in a bidirectional configuration and to study the influence of the thickness of the layers in a given configuration. In both cases, the reflectance spectra and the trichromatic coordinates are calculated and compared. PMID- 17912293 TI - Performance of the holographic multivergence target in the subjective measurement of spherical refractive error and amplitude of accommodation of the human eye. AB - We recently suggested the use of a holographic multivergence target to measure the spherical refractive error and the amplitude of accommodation of the human eye [K. V. Avudainayagam and C. S. Avudainayagam, Opt. Lett.28, 123 (2003)]. In this paper we report the performance of the holographic target in measuring real eyes. The holographic technique compared well with subjective refraction and autorefraction in the measurement of spherical refractive error. The performance of the holographic technique in measuring the amplitude of accommodation was similar to that of the minus lens to blur method and that of the push-up method. These results promote holography as a promising technique for testing human vision. PMID- 17912296 TI - Polarization changes of partially coherent pulses propagating in optical fibers. AB - We consider polarization changes of partially coherent pulses propagating through birefringent dispersive fibers in the linear regime. We show that the evolution of the degree of polarization across such pulses is determined not only by the coherence properties of the pulse in the source plane, but also by the spatial walk-off introduced by the group-velocity mismatch between the two polarization components. The interplay between these two factors determines the asymptotic value of the degree of polarization of an initially unpolarized statistical pulse. We compare our results with previously discussed coherence-induced polarization changes of partially coherent beams propagating in free space. PMID- 17912295 TI - Focus detection from digital in-line holograms based on spectral l1 norms. AB - A rapid focus-detection technique based directly on the spectral content of digital holograms is developed. It differs from previous approaches in that it does not need a full reconstruction of the image. The technique uses l(1) norms of object spectral components associated with the real and imaginary parts of the reconstruction kernel. Further, the l(1) norms can be computed efficiently in the spatial frequency domain using a polar coordinate system, yielding a drastic speedup of approximately 2 orders of magnitude compared with image-based focus detection. Significant computational savings are achieved when subsequent image reconstructions are done selectively over the detected focus distances. Focus detection results from holograms of plankton are demonstrated that show the technique is both accurate and robust. PMID- 17912297 TI - Performance of diffractive optical elements for homogenizing partially coherent light. AB - The effectiveness of different types of diffractive optical element (DOE) for homogenizing partially coherent beams is analyzed, both analytically and numerically. The effectiveness is described by the homogenizing parameter, defined as the inverse of the normalized variance of the dose distribution. For an important class of DOEs designed with common discrete-Fourier-transform methods, it is found that the homogenizing parameter is only of the order of the number of coherence cells in the illuminating beam. However, for a different type of DOE that produces distinct beams under coherent illumination, the homogenizing parameter can be an order of magnitude higher. The inherent dehomogenizing effect caused by the limited temporal duration of the beam, a phenomenon sometimes referred to as dynamic speckle, is also considered. PMID- 17912299 TI - Direct reconstruction of complex refractive index distribution from boundary measurement of intensity and normal derivative of intensity. AB - We present an optical tomographic reconstruction method to recover the complex refractive index distribution from boundary measurements based on intensity, which are the logarithm of intensity and normal derivative of intensity. The method, which is iterative, repeatedly implements the forward propagation equation for light amplitude, the Helmholtz equation, and computes appropriate sensitivity matrices for these measurements. The sensitivity matrices are computed by solving the forward propagation equation for light and its adjoint. The results of numerical experiments show that the data types ln(I) and partial differential I/ partial differential n reconstructed, respectively, the imaginary and the real part of the object refractive index distribution. Moreover, the imaginary part of the refractive index reconstructed from partial differential I/ partial differential n and the real part from ln(I) failed to show the object's inhomogeneity. The value of the propagation constant, k, used in our simulations was 50, and this value resulted in smoothing of the reconstructed inhomogeneities. Thus we have shown that it is possible to reconstruct the complex refractive index distribution directly from the measured intensity without having to first find the light amplitude, as is done in most of the currently available reconstruction algorithms of diffraction tomography. PMID- 17912298 TI - Corneal minimal visible lesion thresholds for 2.0 microm laser radiation. AB - To support refinement of the ANSI Maximum Permissible Exposure safety limits, a series of experiments were conducted in vivo on Dutch Belted rabbit corneas to determine corneal minimum visible lesion thresholds for 2.0 microm continuous wave laser irradiation. Single pulse radiant exposures were made at specified pulse durations of 0.1, 0.25, 0.5, 1.0, 2.0, and 4.0 s for spot 1/e(2) diameters of 1.17 mm and 4.02 mm. Threshold lesions were defined as the presence of a superficial surface whitening one hour after irradiation. Temperature measurements indicated that threshold peak temperatures were dependent on spot size and exposure duration. The exposure duration dependence of threshold average radiant exposure was described by an empirical power law equation: threshold radiant exposure[J/cm(2)]=a x exposure duration[s](b). PMID- 17912300 TI - Modal method for conical diffraction on a rectangular slit metallic grating in a multilayer structure. AB - The modal method is applied to the problem of conical diffraction on a rectangular slit metallic grating lying on an arbitrary multilayer medium. In the approximation of the surface impedance boundary condition on the grating walls, a single matrix equation is obtained, whose coefficients are expressed simply by the reflectivities on the different layers. A simple and comprehensive treatment is thus obtained for virtually any multilayer system. The method is illustrated for the case of a cavity formed by a planar metallic mirror and a grating, as well as the system formed by a doped layer with Drude susceptibility in a substrate below the grating. The method could be useful for the design of near- and far-infrared devices. PMID- 17912301 TI - Monochromatic electromagnetic fields with maximum focal energy density. AB - The monochromatic electromagnetic fields that achieve maximum focal energy density for a given input power and directional spread are found through a variational approach. It is found that the polarization of the fields at the focal point must be perpendicular to the main direction of propagation. Parametric expressions relating the directional spread and focal energy density of these fields (and others that achieve stationary focal irradiance) are found, both in the optical (electric field only) and in the electromagnetic cases. PMID- 17912302 TI - Rigorous electromagnetic analysis of volumetrically complex media using the slice absorption method. AB - There is tremendous demand for numerical methods to perform rigorous analysis of devices that are both large scale and complex throughout their volume. This can arise when devices must be considered with realistic geometry or when they contain artificial materials such as photonic crystals, left-handed materials, nanoparticles, or other metamaterials. The slice absorption method (SAM) was developed to address this need. The method is fully numerical and able to break large problems down into small pieces, or slices, using matrix division or Gaussian elimination instead of eigensystem computations and scattering matrix manipulations. In these regards, the SAM is an attractive alternative to popular techniques like the finite-difference time domain method, rigorous coupled-wave analysis, and the transfer matrix method. To demonstrate the utility of the SAM and benchmark its accuracy, reflection was simulated for a photonic crystal fabricated in SU-8 by multiphoton direct laser writing. Realistic geometry was incorporated into the model by simulating the microfabrication process, which yielded simulation results that matched experimental measurements remarkably well. PMID- 17912303 TI - Experimental implementation of the gyrator transform. AB - The gyrator transform (GT) promises to be a useful tool in image processing, holography, beam characterization, mode transformation, and quantum information. We introduce what we believe to be the first flexible optical experimental setup that performs the GT for a wide range of transformation parameters. The feasibility of the proposed scheme is demonstrated on the gyrator transformation of Hermite-Gaussian modes. For certain parameters the output mode corresponds to the Laguerre-Gaussian one. PMID- 17912304 TI - Sidelobe suppression in chiral optical filters by apodization of the local form birefringence. AB - Chiral optical filters are characterized by circular Bragg effects, including the preferential reflection and transmission of circular polarization states. The selective response to circularly polarized light is caused by stratified birefringent plates twisted into a helical arrangement, as seen in cholesteric liquid crystals and columnar thin films produced by oblique-angle physical vapor deposition. A refinement of the latter, glancing angle deposition employs substrate rotation to control the optical anisotropy of columnar thin films, and was used in this study to suppress the reflection sidelobes of chiral optical filters by modulating the local birefringence of helically structured thin films using an apodization function. Both theoretical simulations based on Berreman formalism, and experimental results involving evaporated TiO(2) thin films are presented and compared. PMID- 17912305 TI - Intensity-ratio error compensation for triangular-pattern phase-shifting profilometry. AB - We present an intensity-ratio error-compensation method to decrease the measurement error caused by projector gamma nonlinearity and image defocus in triangular-pattern phase-shifting profilometry. The intensity-ratio measurement error is first determined by simulating the measurement with the triangular pattern phase-shifting method with ideal and real captured triangular-pattern images based on the ideal and real gamma nonlinearity functions. A lookup table that stores the intensity-ratio measurement error corresponding to the measured intensity ratio is constructed and used for intensity-ratio error compensation. Experiments demonstrated that the intensity-ratio error compensation method significantly reduced the measurement error in the triangular-pattern phase shifting method by 28.5%. PMID- 17912306 TI - Scattering of a Gaussian beam by an impedance half-plane. AB - The diffraction of a Gaussian beam by an impedance half-plane is studied through the method of the modified theory of physical optics. An electric line source, which is defined in the complex space, is used to represent the Gaussian beam. The uniform evaluation of the diffraction integral is performed and the scattering patterns of the field are investigated for various numerical parameters of the incident wave. PMID- 17912307 TI - Perceived shifts in saturation and hue of chromatic stimuli in the near peripheral retina. AB - Using an asymmetric color matching technique, we measured the perceived changes that occur in the saturation and hue of colored stimuli at different eccentricities within the central 25 degrees of the human retina in nine color normal subjects. A cone-opponent-based vector model was used to compute the activity of the L-M and S-(L+M) channels. The results show that a large proportion of the shifts in saturation and hue that occur with increasing retinal eccentricity are mirrored by decreased activity of the L-M channel. In comparison, the contribution of the S cone-opponent system undergoes relatively little change within the central 20 degrees . In addition, we also found that changes in saturation and hue are different from each other in terms of their variation across color space and their variation with stimulus size. Our findings suggest that perceived shifts in saturation and hue are mediated largely via the reduction in activation of the L-M cone-opponent channel but that saturation and hue might be subject to different retinal and/or cortical influences that contribute to their differing size dependencies in the peripheral retina. PMID- 17912308 TI - Effects of Fourier-plane amplitude and phase errors on image reconstruction. I. Small amplitude errors. AB - The effects of Fourier-plane, or spectral, amplitude and phase errors on reconstructed images are studied in terms of the expected mean square error in the image. The relationship between the variance of amplitude and phase errors and the expected mean square error is derived for small amplitude errors and arbitrarily large phase errors. This allows "equivalent" amplitude and phase errors to be defined. The effects of large amplitude errors are discussed in general terms. Simulations are used to verify these relationships, and the effects of spectral amplitude and phase errors on reconstructed images are compared. PMID- 17912309 TI - Noninterferometric characterization of partially coherent scalar wave fields and application to scattered light. AB - We report on the application of a simple propagation-based phase-space tomographic technique to the determination of characteristic projections through the mutual optical intensity and the generalized radiance of a scalar, quasi monochromatic partially coherent wave field. This method is applied to the reconstruction of the coherence functions of an initially spatially coherent optical wave field that has propagated through a suspension of polystyrene microspheres. As anticipated, we see that the field separates into a ballistic, or unscattered, component and a scattered component with a much shorter coherence length. Good agreement is obtained between experimental results and the results of a model based on a wave-transport equation. PMID- 17912310 TI - Inferring path average Cn2 values in the marine environment. AB - Current mathematical scintillation theory describing laser propagation through the atmosphere has been developed for terrestrial environments. Scintillation expressions valid in all regimes of optical turbulence for propagation in the maritime environment, based on what we believe to be a newly developed marine atmospheric spectrum, have been developed for spherical waves. Path average values of the structure parameter, C(n)(2), were inferred from optical scintillation measurements of a diverged laser beam propagating in a marine environment, using scintillation expressions based on both terrestrial and marine refractive index spectra. In the moderate-to-strong fluctuation regime, the inferred marine C(n)(2) values were about 20% smaller than inferred terrestrial C(n)(2) values, but a minimal difference was observed in the weak fluctuation regime. Measurements of angle-of-arrival fluctuations were used to infer C(n)(2) values in the moderate-to-strong fluctuation regime, resulting in values of the structure parameter that were at least an order of magnitude larger than the two scintillation-inferred C(n)(2) values. PMID- 17912311 TI - Observation of light transmission through randomly rough glass surfaces beyond the critical angle. AB - In this series of experiments we study the transmission of laser light through a randomly rough interface between air and ground-glass diffusers. We verify the refractive index suppression predicted by Dogariu and Boreman [Opt. Lett.21, 701 (1996)]. We also observe and quantify transmission beyond the critical angle for total internal reflection defined by Snell's law. In particular, these results may be applied to the ANITA experiment, which detects astrophysical neutrino interactions via radio waves produced under the ice. These radio waves must pass through the rough surfaces of the Antarctic ice sheets and shelves. PMID- 17912312 TI - Recovery of spectral reflectances of objects being imaged by multispectral cameras. AB - Acquisition of spectral information of objects being imaged through the use of sensor responses is important to reproduce color images under various illuminations. In the past several models have been proposed to recover the spectral reflectances from sensor responses. The accuracy of the spectral reflectances recovered by five different models is compared by using multispectral cameras. It is shown that the Wiener estimation that uses the noise variance estimated as proposed in IEEE Trans. Image Process.15, 1848 (2006) recovers the spectral reflectances more accurately than the others when the test samples are different from learning samples. PMID- 17912313 TI - Transversal superresolution with noncontact axial movement of periodic structures. AB - We present an innovative approach that allows superresolved images to be obtained by axial moving of two gratings and time integrating in the detector plane. The two gratings do not have to be in contact with either the object or the detector, and both are positioned between the object and the image planes. One of the main applications for the proposed approach in contrast to previously discussed time multiplexing superresolving methods is that it may fit well to superresolved imaging of remote objects, since both gratings are not in contact with either the object or the detector planes. PMID- 17912314 TI - Diffraction of evanescent plane waves by a resistive half-plane. AB - Diffraction of evanescent plane waves by a resistive half-plane is examined. The scattering integrals are constructed with the modified theory of physical optics. These integrals are evaluated uniformly by using an unusual method. The scattered fields of evanescent waves are obtained by giving the angle of incidence a complex value. The diffracted waves are plotted numerically for different parameters of the incident field. PMID- 17912315 TI - Some observations on the masking effects of Mach bands. AB - There are 8 cycle/deg ripples or oscillations in performance as a function of location near Mach bands in experiments measuring Mach bands' masking effects on random polarity signal bars. The oscillations with increments are 180 degrees out of phase with those for decrements. The oscillations, much larger than the measurement error, appear to relate to the weighting function of the spatial frequency-tuned channel detecting the broadband signals. The ripples disappear with step maskers and become much smaller at durations below 25 ms, implying either that the site of masking has changed or that the weighting function and hence spatial-frequency tuning is slow to develop. PMID- 17912316 TI - Role of optics in the accuracy of depth-from-defocus systems: comment. AB - In their paper "Role of optics in the accuracy of depth-from-defocus systems" [J. Opt. Soc. Am. A24, 967 (2007)] the authors Blayvas, Kimmel, and Rivlin discuss the effect of optics on the depth reconstruction accuracy. To this end they applied an approach in Fourier space. An alternative derivation of their result in the spatial domain, based on geometrical optics, is presented and compared with their outcome. A better agreement with experimental data is achieved if some unclarities are refined. PMID- 17912317 TI - Wind profiling based on the optical beam intensity statistics in a turbulent atmosphere. AB - Reconstruction of the wind profile from the statistics of intensity fluctuations of an optical beam propagating in a turbulent atmosphere is considered. The equations for the spatiotemporal correlation function and the spectrum of weak intensity fluctuations of a Gaussian beam are obtained. The algorithms of wind profile retrieval from the spatiotemporal intensity spectrum are described and the results of end-to-end computer experiments on wind profiling based on the developed algorithms are presented. It is shown that the developed algorithms allow retrieval of the wind profile from the turbulent optical beam intensity fluctuations with acceptable accuracy in many practically feasible laser measurements set up in the atmosphere. PMID- 17912318 TI - Embedded centrosymmetric multilayer stacks as complete-transmission quarter-wave and half-wave retarders under conditions of frustrated total internal reflection. AB - A centrosymmetric multilayer stack of two transparent materials, which is embedded in a high-index prism, can function as a complete-transmission quarter wave or half-wave retarder (QWR or HWR) under conditions of frustrated total internal reflection. The multilayer consists of a high-index center layer sandwiched between two identical low-index films with high-index-low-index bilayers repeated on both sides of the central trilayer, maintaining the symmetry of the entire stack and constituting a QWR (Delta(t)=90 degrees or 270 degrees ) or HWR (Delta(t)=180 degrees ) in transmission. A QWR design at wavelength lambda=1.55 microm is presented that employs an 11-layer stack of Si and SiO(2) thin films, which is embedded in a GaP cube prism. The intensity transmittances for the p and s polarizations remain >99% and Delta(t) deviates from 90 degrees by <+/-3 degrees over a 100 nm spectral bandwidth (1.5< or =lambda< or =1.6 microm), and by < or =+/-7 degrees over an internal field view of +/-1 degrees (incidence angle 44 degrees < or = phi(0)< or =46 degrees inside the prism). An HWR design at lambda=1.55 microm employs seven layers of Si and SiO(2) thin films embedded in a Si cube, has an average transmittance >93%, and Delta(t) that differs from 180 degrees by <+/-0.3 degrees over a 100 nm bandwidth (1.5< or =lambda< or =1.6 microm) and by <+/-17 degrees over an internal field view of +/ 1 degree . The sensitivity of these devices to film-thickness errors is also considered. PMID- 17912319 TI - Optimizing frequency-domain fluorescence lifetime sensing for high-throughput applications: photon economy and acquisition speed. AB - The signal-to-noise ratio of a measurement is determined by the photon economy of the detection technique and the available photons that are emitted by the sample. We investigate the efficiency of various frequency-domain lifetime detection techniques also in relation to time-domain detection. Nonlinear effects are discussed that are introduced by the use of image intensifiers and by fluorophore saturation. The efficiency of fluorescence lifetime imaging microscopy setups is connected to the speed of acquisition and thus to the imaging throughput. We report on the optimal conditions for balancing signal-to-noise ratio and acquisition speed in fluorescence lifetime sensing. PMID- 17912320 TI - Mechanism of compensation of aberrations in the human eye. AB - We studied the mechanism of compensation of aberrations within the young human eye by using experimental data and advanced ray-tracing modeling. Corneal and ocular aberrations along with the alignment properties (angle kappa, lens tilt, and decentration) were measured in eyes with different refractive errors. Predictions from individualized ray-tracing optical models were compared with the actual measurements. Ocular spherical aberration was, in general, smaller than corneal spherical aberration without relation to refractive error. However, horizontal coma compensation was found to be significantly larger for hyperopic eyes where angle kappa tended to also be larger. We propose a simple analytical model of the relationship between the corneal coma compensation effect with the field angle and corneal and crystalline shape factors. The actual shape factors corresponded approximately to the optimum shapes that automatically provide this coma compensation. We showed that the eye behaves as an aplanatic optical system, an optimized design solution rendering stable retinal image quality for different ocular geometries. PMID- 17912321 TI - Experimental reconstruction of a highly reflecting fiber Bragg grating by using spectral regularization and inverse scattering. AB - We demonstrate experimentally, for the first time to our knowledge, a reconstruction of a highly reflecting fiber Bragg grating from its complex reflection spectrum by using a regularization algorithm. The regularization method is based on correcting the measured reflection spectrum at the Bragg zone frequencies and enables the reconstruction of the grating profile using the integral-layer-peeling algorithm. A grating with an approximately uniform profile and with a maximum reflectivity of 99.98% was accurately reconstructed by measuring only its complex reflection spectrum. PMID- 17912322 TI - Phase retrieval and saddle-point optimization. AB - Iterative algorithms with feedback are among the most powerful and versatile optimization methods for phase retrieval. Among these, the hybrid input-output algorithm has demonstrated practical solutions to giga-element nonlinear phase retrieval problems, escaping local minima and producing images at resolutions beyond the capabilities of lens-based optical methods. Here the input-output iteration is improved by a lower-dimensional subspace saddle-point optimization. PMID- 17912323 TI - Propagating free-space nonparaxial beams. AB - We use a method based on the simultaneous combination of the propagation operator and the Fourier transform with arbitrary index in propagating the transverse component of a nonparaxial beam in free space from an arbitrary initial transverse field structure. Being an iterative method, this approach can easily be implemented computationally. As an example of its efficiency, we derive the closed-form nonparaxial corrections to a Bessel-Gaussian beam, showing that our results differ strongly from those reported previously. The validity of our approach is supported by an analysis of the paraxiality estimator recently introduced in the literature. PMID- 17912324 TI - Integrated liquid-crystal switch for both TE and TM modes: proposal and design. AB - An integrated optical switch is proposed and designed based on a weak-anchoring liquid-crystal (LC) cell with a substrate integrating planar lightwave circuits. It consists of a polarization splitter, two switchable polarization converters and a polarization combiner. The polarization splitter/combiner is a directional coupler with an etched slot and filled-in LC covering layer. The switchable polarization converters are straight waveguides with a designed length and covering LC. The proposed configuration is superior to the existing integrated LC switches, as it works for both the TE and TM modes. PMID- 17912325 TI - Application of the multiple-scattering method to analysis of systems with semi infinite photonic waveguides. AB - We propose a technique of compensating the spurious reflections implied by the multiple-scattering (MS) method, commonly used for analyzing finite photonic crystal (PC) systems, to obtain exact values of characteristic parameters, such as reflection and transmission coefficients, of PC functional elements. Rather than a modification of the MS computational algorithm, our approach involves postprocessing of results obtained by the MS method. We derive analytical formulas for the fields excited in a finite system, taking explicitly into account the spurious reflections occurring at the artificial system boundaries. The intrinsic parameters of the investigated functional element are found by fitting the results of MS simulations to those obtained from the formulas derived. Devices linked with one and two semi-infinite waveguides are analyzed explicitly; possible extensions of the formalism to more complex circuits are discussed as well. The accuracy of the proposed method is tested in a number of systems. The results of our calculations prove to be in good agreement with those obtained independently by other authors. PMID- 17912326 TI - Application of the multiscale singular perturbation method to nonparaxial beam propagations in free space. AB - Starting from the vector Maxwell's equations and applying the multiscale singular perturbation method, the nonparaxial beam propagation is studied in free space. Two new equations have been derived for transverse and longitudinal electric fields of an arbitrary polarized electromagnetic wave even in the case of tightly focused nonparaxial laser beams. By using the analogy of the optical beam in the space domain and the optical pulse in the time domain, the higher-order diffraction term is introduced. For strongly nonparaxial beams that are characterized by large values of the perturbative parameter, our correction solutions yield an accurate description of the field in the near-field region and are consistent with all other correction results obtained by others in the far field region. For weakly nonparaxial beams, our correction solutions can be expressed in a very simple form that is proved to be exactly consistent with the solutions obtained by Cao and Deng [J. Opt. Soc. Am. A15, 1144 (1998)]. In addition, the lowest-order correction to the paraxial approximation can be found to be in good agreement with the results of Lax et al. [Phys. Rev. A11, 1365 (1975)] and Seshadri [J. Opt. Soc. Am. A19, 2134 (2002)]. PMID- 17912327 TI - Analysis of photometric factors based on photometric linearization. AB - We propose a method for analyzing photometric factors, such as diffuse reflection, specular reflection, attached shadow, and cast shadow. For analyzing real images, we utilize the photometric linearization method, which was originally proposed for image synthesis. First, we show that each pixel can be photometrically classified by a simple comparison of the pixel intensity. Our classification algorithm requires neither 3D shape information nor color information of the scene. Then, we show that the accuracy of the photometric linearization can be improved by introducing a new classification-based criterion to the linearization process. Experimental results show that photometric factors can be correctly classified without any special devices. A further experiment shows that the proposed method is effective for photometric stereo. PMID- 17912329 TI - Communication modes with partially coherent fields. AB - We develop a theory for the description of partially coherent wave fields in linear optical systems in terms of the so-called communication modes. The communication modes are the singular functions and singular values of the appropriate propagation kernels. In particular, we show that optical fields of any state of coherence may be readily propagated through deterministic systems using the modal representation based on the system properties. The relation of the communication modes to the conventional coherent-mode representation is discussed, and expressions for the effective degree of coherence in the optical system are derived. The results are illustrated by numerical examples in optical near-field geometry. PMID- 17912331 TI - Radiation pattern of azimuthally varying scalar Laguerre-Gauss waves. AB - The exact full-wave generalization of the azimuthally varying scalar real argument Laguerre-Gauss beam is obtained. The radiation intensity of the resulting azimuthally varying real-argument Laguerre-Gauss wave is determined. The main characteristics of the radiation intensity pattern are discussed. The total power P(n,m) is determined, where n is the radial mode number and m is the azimuthal mode number. By the use of 1/P(n,m), the characteristics of the quality of the paraxial beam approximation to the real-argument Laguerre-Gauss wave are investigated. PMID- 17912330 TI - Absorption of ultrashort optical pulses in water. AB - We investigate the linear propagation of 800 and 1530 nm ultrashort optical pulses in water. For all pulse repetition rates studied, we observe pure exponential decay down to a transmission of 2.5 x 10(-5). We further demonstrate that previous observations of nonmonoexponential decay and pulse splitup in broadband pulses are consistent with Beer's law in the purely linear regime. PMID- 17912332 TI - Phase retrieval from speckle images. AB - In ground-based astronomy, the inverse problem of phase retrieval from speckle images is a means to calibrate static aberrations for correction by active optics. It can also be used to sense turbulent wavefronts. However, the number of local minima drastically increases with the turbulence strength, mainly because of phase wrapping ambiguities. Multifocal phase diversity has been considered to overcome some ambiguities of the phase retrieval problem. We propose an effective algorithm for phase retrieval from a single focused image. Our algorithm makes use of a global optimization strategy and an automatically tuned smoothness prior to overcome local minima and phase degeneracies. We push the limit of D/r(0)=4 achieved by Irwan and Lane [J. Opt. Soc. Am. A.15, 2302 (1998)] up to D/r(0)=11, which is a major improvement owing to the drastic increase in the problem complexity. We estimate the performances of our approach from consistent simulations for different turbulence strengths and noise levels (down to 1500 photons per image). We also investigate the benefit of temporal correlation. PMID- 17912333 TI - Effect of beam size parameters on internal fields in an infinite cylinder irradiated by an elliptical Gaussian beam. AB - The scattered and internal fields of an infinite, homogeneous cylinder illuminated by a linearly polarized beam depend on the following parameters: the object size parameter of the cylinder (ka, where k=2pi/lambda, lambda is the wavelength of the incident beam in the surrounding medium, and a is the radius of cylinder), the complex relative refractive index of the object, the beam size parameters (komega(1) and komega(2), where omega(1), omega(2) are the representative beam dimensions), the angle between the cylinder axis and the Poynting vector of the incident wave, and the angle between the plane of polarization and the plane of incidence. Only when the dimensions of the beam are much greater than the cylinder diameter, and hence the portion of the beam interacting with the cylinder is essentially uniform, can the plane-wave solution be used in computing the scattered and internal fields. Hence a rigorous electromagnetic approach like the generalized Lorenz-Mie theory for spheres is used to study the effect of beam size parameters on the internal fields in an infinite cylinder irradiated by elliptical Gaussian beams. The significant effects of beam size parameters on the internal fields in an infinite cylinder are presented using specific cases of (1) resonance effects in a glass cylinder (ka=45.726, transverse-electric mode 53,3) and (2) a cylindrical microchannel (ka approximately 760) irradiated by a 632.8 nm laser beam. PMID- 17912334 TI - Unique-hue stimulus selection using Munsell color chips. AB - Presented are intra- and inter-observer variability data comparing the unique-hue (UH) selections of sets of males and females, using two different visual experimental procedures incorporating Munsell color chips of varying hue but identical chroma and value. Although 34 of the 40 Munsell hue chips were selected by at least one observer as a UH, selections were generally repeatable. In addition, intra-observer variability represented approximately 15% of inter observer variability. Also, when only three consecutive Munsell chips were viewed at a time, females showed significantly larger intra-observer variability than males, especially when making unique green selections. However, variability in UH selections was statistically insignificant between males and females when all Munsell chips were viewed simultaneously. No correlation was found between UH selections or intra-observer variability and hue ordering ability. PMID- 17912335 TI - Nonparaxial analysis of continuous self-imaging gratings in oblique illumination. AB - Tolerance in angles of continuously self-imaging gratings (CSIGs) is explored. The degradation in angle of the shape of the point-spread function is theoretically investigated and illustrated by simulations and experiments. The formalism presented is inspired by the one used for classical lenses and can be easily generalized to diffraction gratings. It turns out that well-designed CSIGs could be used for scanning optical systems requiring a large field of view. PMID- 17912336 TI - Stokes-vector evolution in a weakly anisotropic inhomogeneous medium. AB - The equation for evolution of the four-component Stokes vector in weakly anisotropic and smoothly inhomogeneous media is derived on the basis of a quasi isotropic approximation of the geometrical optics method, which provides the consequent asymptotic solution of Maxwell's equations. Our equation generalizes previous results obtained for the normal propagation of electromagnetic waves in stratified media. It is valid for curvilinear rays with torsion and is capable of describing normal mode conversion in inhomogeneous media. Remarkably, evolution of the four-component Stokes vector is described by the Bargmann-Michel-Telegdi equation for relativistic spin precession, whereas the equation for the three component Stokes vector resembles the Landau-Lifshitz equation describing spin precession in ferromagnetic systems. The general theory is applied for analysis of polarization evolution in a magnetized plasma. We also emphasize fundamental features of the non-Abelian polarization evolution in anisotropic inhomogeneous media and illustrate them by simple examples. PMID- 17912337 TI - Comparison of a physical model and principal component analysis for the diagnosis of epithelial neoplasias in vivo using diffuse reflectance spectroscopy. AB - We explored the use of diffuse reflectance spectroscopy in the ultraviolet visible (UV-VIS) spectrum for the diagnosis of epithelial precancers and cancers in vivo. A physical model (Monte Carlo inverse model) and an empirical model (principal component analysis, (PCA)) based approach were compared for extracting diagnostic features from diffuse reflectance spectra measured in vivo from the dimethylbenz[alpha]anthracene-treated hamster cheek pouch model of oral carcinogenesis. These diagnostic features were input into a support vector machine algorithm to classify each tissue sample as normal (n=10) or neoplastic (dysplasia to carcinoma, n=10) and cross-validated using a leave one out method. There was a statistically significant decrease in the absorption and reduced scattering coefficient at 460 nm in neoplastic compared to normal tissues, and these two features provided 90% classification accuracy. The first two principal components extracted from PCA provided a classification accuracy of 95%. The first principal component was highly correlated with the wavelength-averaged reduced scattering coefficient. Although both methods show similar classification accuracy, the physical model provides insight into the physiological and structural features that discriminate between normal and neoplastic tissues and does not require a priori, a representative set of spectral data from which to derive the principal components. PMID- 17912339 TI - Membrane type-1 matrix metalloproteinases and tissue inhibitor of metalloproteinases-2 RNA levels mimic each other during Xenopus laevis metamorphosis. AB - Matrix metalloproteinases (MMPs) and their endogenous inhibitors TIMPs (tissue inhibitors of MMPs), are two protein families that work together to remodel the extracellular matrix (ECM). TIMPs serve not only to inhibit MMP activity, but also aid in the activation of MMPs that are secreted as inactive zymogens. Xenopus laevis metamorphosis is an ideal model for studying MMP and TIMP expression levels because all tissues are remodeled under the control of one molecule, thyroid hormone. Here, using RT-PCR analysis, we examine the metamorphic RNA levels of two membrane-type MMPs (MT1-MMP, MT3-MMP), two TIMPs (TIMP-2, TIMP-3) and a potent gelatinase (Gel-A) that can be activated by the combinatory activity of a MT-MMP and a TIMP. In the metamorphic tail and intestine the RNA levels of TIMP-2 and MT1-MMP mirror each other, and closely resemble that of Gel-A as all three are elevated during periods of cell death and proliferation. Conversely, MT3-MMP and TIMP-3 do not have similar RNA level patterns nor do they mimic the RNA levels of the other genes examined. Intriguingly, TIMP-3, which has been shown to have anti-apoptotic activity, is found at low levels in tissues during periods of apoptosis. PMID- 17912341 TI - Optimality driven nearest centroid classification from genomic data. AB - Nearest-centroid classifiers have recently been successfully employed in high dimensional applications, such as in genomics. A necessary step when building a classifier for high-dimensional data is feature selection. Feature selection is frequently carried out by computing univariate scores for each feature individually, without consideration for how a subset of features performs as a whole. We introduce a new feature selection approach for high-dimensional nearest centroid classifiers that instead is based on the theoretically optimal choice of a given number of features, which we determine directly here. This allows us to develop a new greedy algorithm to estimate this optimal nearest-centroid classifier with a given number of features. In addition, whereas the centroids are usually formed from maximum likelihood estimates, we investigate the applicability of high-dimensional shrinkage estimates of centroids. We apply the proposed method to clinical classification based on gene-expression microarrays, demonstrating that the proposed method can outperform existing nearest centroid classifiers. PMID- 17912342 TI - Heparin-binding-hemagglutinin-induced IFN-gamma release as a diagnostic tool for latent tuberculosis. AB - BACKGROUND: The detection of latent tuberculosis infection (LTBI) is a major component of tuberculosis (TB) control strategies. In addition to the tuberculosis skin test (TST), novel blood tests, based on in vitro release of IFN gamma in response to Mycobacterium tuberculosis-specific antigens ESAT-6 and CFP 10 (IGRAs), are used for TB diagnosis. However, neither IGRAs nor the TST can separate acute TB from LTBI, and there is concern that responses in IGRAs may decline with time after infection. We have therefore evaluated the potential of the novel antigen heparin-binding hemagglutinin (HBHA) for in vitro detection of LTBI. METHODOLOGY AND PRINCIPAL FINDINGS: HBHA was compared to purified protein derivative (PPD) and ESAT-6 in IGRAs on lymphocytes drawn from 205 individuals living in Belgium, a country with low TB prevalence, where BCG vaccination is not routinely used. Among these subjects, 89 had active TB, 65 had LTBI, based on well-standardized TST reactions and 51 were negative controls. HBHA was significantly more sensitive than ESAT-6 and more specific than PPD for the detection of LTBI. PPD-based tests yielded 90.00% sensitivity and 70.00% specificity for the detection of LTBI, whereas the sensitivity and specificity for the ESAT-6-based tests were 40.74% and 90.91%, and those for the HBHA-based tests were 92.06% and 93.88%, respectively. The QuantiFERON-TB Gold In-Tube (QFT IT) test applied on 20 LTBI subjects yielded 50% sensitivity. The HBHA IGRA was not influenced by prior BCG vaccination, and, in contrast to the QFT-IT test, remote (>2 years) infections were detected as well as recent (<2 years) infections by the HBHA-specific test. CONCLUSIONS: The use of ESAT-6- and CFP-10 based IGRAs may underestimate the incidence of LTBI, whereas the use of HBHA may combine the operational advantages of IGRAs with high sensitivity and specificity for latent infection. PMID- 17912343 TI - Novel decapeptides that bind avidly and deliver radioisotope to colon cancer cells. AB - BACKGROUND: The rapidly growing field of targeted tumor therapy often utilizes an antibody, sometimes tagged with a tumor-ablating material such as radioisotope, directed against a specific molecule. METHODOLOGY/PRINCIPAL FINDINGS: This report describes the discovery of nine novel decapeptides which can be radioactively labeled, bind to, and deliver (32)P to colon cancer cells. The decapeptides vary from one another by one to three amino acids and demonstrate vastly different binding abilities. The most avidly binding decapeptide can permanently deliver very high levels of radioisotope to the adenocarcinoma cancer cell lines at an efficiency 35 to 150 times greater than to a variety of other cell types, including cell lines derived from other types of cancer or from normal tissue. CONCLUSIONS/SIGNIFICANCE: This experimental approach represents a new example of a strategy, termed peptide binding therapy, for the potential treatment of colorectal and other adenocarcinomas. PMID- 17912340 TI - Sexual risk factors for HIV infection in early and advanced HIV epidemics in sub Saharan Africa: systematic overview of 68 epidemiological studies. AB - BACKGROUND: It is commonly assumed that sexual risk factors for heterosexual HIV transmission in sub-Saharan Africa, such as multi-partner sex, paid sex and co infections, become less important as HIV epidemics mature and prevalence increases. METHODS AND FINDINGS: We conducted a systematic review of 68 African epidemiological studies from 1986 to 2006 involving 17,000 HIV positive adults and 73,000 controls. We used random-effects methods and stratified results by gender, time, background HIV prevalence rates and other variables. The number of sex partners, history of paid sex, and infection with herpes simplex virus (HSV 2) or other sexually-transmitted infections (STIs) each showed significant associations with HIV infection. Among the general population, the odds ratio (OR) of HIV infection for women reporting 3+ sex partners versus 0-2 was 3.64 (95%CI [2.87-4.62]), with similar risks for men. About 9% of infected women reported ever having been paid for sex, versus 4% of control women (OR = 2.29, [1.45-3.62]). About 31% of infected men reported ever paying for sex versus 18% of uninfected men (OR = 1.75, [1.30-2.36]). HSV-2 infection carried the largest risk of HIV infection: OR = 4.62, [2.85-7.47] in women, and OR = 6.97, [4.68 10.38] in men. These risks changed little over time and stratification by lower and higher HIV background prevalence showed that risk ratios for most variables were larger in high prevalence settings. Among uninfected controls, the male female differences in the number of sex partners and in paid sex were more extreme in the higher HIV prevalence settings than in the lower prevalence settings. SIGNIFICANCE: Multi-partner sex, paid sex, STIs and HSV-2 infection are as important to HIV transmission in advanced as in early HIV epidemics. Even in high prevalence settings, prevention among people with high rates of partner change, such as female sex workers and their male clients, is likely to reduce transmission overall. PMID- 17912344 TI - Transposon excision from an atypical site: a mechanism of evolution of novel transposable elements. AB - The role of transposable elements in sculpting the genome is well appreciated but remains poorly understood. Some organisms, such as humans, do not have active transposons; however, transposable elements were presumably active in their ancestral genomes. Of specific interest is whether the DNA surrounding the sites of transposon excision become recombinogenic, thus bringing about homologous recombination. Previous studies in maize and Drosophila have provided conflicting evidence on whether transposon excision is correlated with homologous recombination. Here we take advantage of an atypical Dissociation (Ds) element, a maize transposon that can be mobilized by the Ac transposase gene in Arabidopsis thaliana, to address questions on the mechanism of Ds excision. This atypical Ds element contains an adjacent 598 base pairs (bp) inverted repeat; the element was allowed to excise by the introduction of an unlinked Ac transposase source through mating. Footprints at the excision site suggest a micro-homology mediated non-homologous end joining reminiscent of V(D)J recombination involving the formation of intra-helix 3' to 5' trans-esterification as an intermediate, a mechanism consistent with previous observations in maize, Antirrhinum and in certain insects. The proposed mechanism suggests that the broken chromosome at the excision site should not allow recombinational interaction with the homologous chromosome, and that the linked inverted repeat should also be mobilizable. To test the first prediction, we measured recombination of flanking chromosomal arms selected for the excision of Ds. In congruence with the model, Ds excision did not influence crossover recombination. Furthermore, evidence for correlated movement of the adjacent inverted repeat sequence is presented; its origin and movement suggest a novel mechanism for the evolution of repeated elements. Taken together these results suggest that the movement of transposable elements themselves may not directly influence linkage. Possibility remains, however, for novel repeated DNA sequences produced as a consequence of transposon movement to influence crossover in subsequent generations. PMID- 17912346 TI - SLiMFinder: a probabilistic method for identifying over-represented, convergently evolved, short linear motifs in proteins. AB - BACKGROUND: Short linear motifs (SLiMs) in proteins are functional microdomains of fundamental importance in many biological systems. SLiMs typically consist of a 3 to 10 amino acid stretch of the primary protein sequence, of which as few as two sites may be important for activity, making identification of novel SLiMs extremely difficult. In particular, it can be very difficult to distinguish a randomly recurring "motif" from a truly over-represented one. Incorporating ambiguous amino acid positions and/or variable-length wildcard spacers between defined residues further complicates the matter. METHODOLOGY/PRINCIPAL FINDINGS: In this paper we present two algorithms. SLiMBuild identifies convergently evolved, short motifs in a dataset of proteins. Motifs are built by combining dimers into longer patterns, retaining only those motifs occurring in a sufficient number of unrelated proteins. Motifs with fixed amino acid positions are identified and then combined to incorporate amino acid ambiguity and variable length wildcard spacers. The algorithm is computationally efficient compared to alternatives, particularly when datasets include homologous proteins, and provides great flexibility in the nature of motifs returned. The SLiMChance algorithm estimates the probability of returned motifs arising by chance, correcting for the size and composition of the dataset, and assigns a significance value to each motif. These algorithms are implemented in a software package, SLiMFinder. SLiMFinder default settings identify known SLiMs with 100% specificity, and have a low false discovery rate on random test data. CONCLUSIONS/SIGNIFICANCE: The efficiency of SLiMBuild and low false discovery rate of SLiMChance make SLiMFinder highly suited to high throughput motif discovery and individual high quality analyses alike. Examples of such analyses on real biological data, and how SLiMFinder results can help direct future discoveries, are provided. SLiMFinder is freely available for download under a GNU license from http://bioinformatics.ucd.ie/shields/software/slimfinder/. PMID- 17912345 TI - A diverse group of previously unrecognized human rhinoviruses are common causes of respiratory illnesses in infants. AB - BACKGROUND: Human rhinoviruses (HRVs) are the most prevalent human pathogens, and consist of 101 serotypes that are classified into groups A and B according to sequence variations. HRV infections cause a wide spectrum of clinical outcomes ranging from asymptomatic infection to severe lower respiratory symptoms. Defining the role of specific strains in various HRV illnesses has been difficult because traditional serology, which requires viral culture and neutralization tests using 101 serotype-specific antisera, is insensitive and laborious. METHODS AND FINDINGS: To directly type HRVs in nasal secretions of infants with frequent respiratory illnesses, we developed a sensitive molecular typing assay based on phylogenetic comparisons of a 260-bp variable sequence in the 5' noncoding region with homologous sequences of the 101 known serotypes. Nasal samples from 26 infants were first tested with a multiplex PCR assay for respiratory viruses, and HRV was the most common virus found (108 of 181 samples). Typing was completed for 101 samples and 103 HRVs were identified. Surprisingly, 54 (52.4%) HRVs did not match any of the known serotypes and had 12-35% nucleotide divergence from the nearest reference HRVs. Of these novel viruses, 9 strains (17 HRVs) segregated from HRVA, HRVB and human enterovirus into a distinct genetic group ("C"). None of these new strains could be cultured in traditional cell lines. CONCLUSIONS: By molecular analysis, over 50% of HRV detected in sick infants were previously unrecognized strains, including 9 strains that may represent a new HRV group. These findings indicate that the number of HRV strains is considerably larger than the 101 serotypes identified with traditional diagnostic techniques, and provide evidence of a new HRV group. PMID- 17912347 TI - Molecular analysis of a leprosy immunotherapeutic bacillus provides insights into Mycobacterium evolution. AB - BACKGROUND: Evolutionary dynamics plays a central role in facilitating the mechanisms of species divergence among pathogenic and saprophytic mycobacteria. The ability of mycobacteria to colonize hosts, to proliferate and to cause diseases has evolved due to its predisposition to various evolutionary forces acting over a period of time. Mycobacterium indicus pranii (MIP), a taxonomically unknown 'generalist' mycobacterium, acts as an immunotherapeutic against leprosy and is approved for use as a vaccine against it. The large-scale field trials of this MIP based leprosy vaccine coupled with its demonstrated immunomodulatory and adjuvant property has led to human clinical evaluations of MIP in interventions against HIV-AIDS, psoriasis and bladder cancer. MIP, commercially available as 'Immuvac', is currently the focus of advanced phase III clinical trials for its antituberculosis efficacy. Thus a comprehensive analysis of MIP vis-a-vis evolutionary path, underpinning its immanent immunomodulating properties is of the highest desiderata. PRINCIPAL FINDINGS: Genome wide comparisons together with molecular phylogenetic analyses by fluorescent amplified fragment length polymorphism (FAFLP), enterobacterial repetitive intergenic consensus (ERIC) based genotyping and candidate orthologues sequencing revealed that MIP has been the predecessor of highly pathogenic Mycobacterium avium intracellulare complex (MAIC) that did not resort to parasitic adaptation by reductional gene evolution and therefore, preferred a free living life-style. Further analysis suggested a shared aquatic phase of MAIC bacilli with the early pathogenic forms of Mycobacterium, well before the latter diverged as 'specialists'. CONCLUSIONS/SIGNIFICANCE: This evolutionary paradigm possibly affirms to marshall our understanding about the acquisition and optimization of virulence in mycobacteria and determinants of boundaries therein. PMID- 17912348 TI - The mucosae-associated epithelial chemokine (MEC/CCL28) modulates immunity in HIV infection. AB - BACKGROUND: CCL28 (MEC) binds to CCR3 and CCR10 and recruits IgA-secreting plasma cells (IgA-ASC) in the mucosal lamina propria (MLP). Mucosal HIV-specific IgA are detected in HIV-infection and exposure. The CCL28 circuit was analyzed in HIV infected and-exposed individuals and in HIV-unexposed controls; the effect of CCL28 administration on gastrointestinal MLP IgA-ASC was verified in a mouse model. METHODOLOGY/FINDINGS: CCL28 was augmented in breast milk (BM) plasma and saliva of HIV-infected and -exposed individuals; CCR3+ and CCR10+ B lymphocytes were increased in these same individuals. Additionally: 1) CCL28 concentration in BM was associated with longer survival in HIV vertically-infected children; and 2) gastro-intestinal mucosal IgA-ASC were significantly increased in VSV immunized mice receiving CCL28. CONCLUSIONS: CCL28 mediates mucosal immunity in HIV exposure and infection. CCL28-including constructs should be considered in mucosal vaccines to prevent HIV infection of the gastro-intestinal MLP via modulation of IgA-ASC. PMID- 17912349 TI - A novel regulator couples sporogenesis and trehalose biogenesis in Aspergillus nidulans. AB - Trehalose is a compatible osmolyte produced by bacteria, fungi, insects and plants to protect the integrity of cells against various environmental stresses. Spores, the reproductive, survival and infection bodies of fungi require high amounts of trehalose for long-term survival. Here, via a gain-of-function genetic screen, we identify the novel regulator VosA that couples the formation of spores and focal trehalose biogenesis in the model fungus Aspergillus nidulans. The vosA gene is expressed specifically during the formation of both sexual and asexual spores (conidia). Levels of vosA mRNA and protein are high in both types of spore. The deletion of vosA results in the lack of trehalose in spores, a rapid loss of the cytoplasm, organelles and viability of spores, and a dramatic reduction in tolerance of conidia to heat and oxidative stress. Moreover, the absence of vosA causes uncontrolled activation of asexual development, whereas the enhanced expression of vosA blocks sporulation, suggesting that VosA also functions in negative-feedback regulation of sporogenesis. VosA localizes in the nucleus of mature conidia and its C-terminal region contains a potential transcription activation domain, indicating that it may function as a transcription factor primarily controlling the late process of sporulation including trehalose biogenesis. VosA is conserved in most fungi and may define a new fungus-specific transcription factor family. PMID- 17912350 TI - Modulation of motor cortex excitability by physical similarity with an observed hand action. AB - The passive observation of hand actions is associated with increased motor cortex excitability, presumably reflecting activity within the human mirror neuron system (MNS). Recent data show that in-group ethnic membership increases motor cortex excitability during observation of culturally relevant hand gestures, suggesting that physical similarity with an observed body part may modulate MNS responses. Here, we ask whether the MNS is preferentially activated by passive observation of hand actions that are similar or dissimilar to self in terms of sex and skin color. Transcranial magnetic stimulation-induced motor evoked potentials were recorded from the first dorsal interosseus muscle while participants viewed videos depicting index finger movements made by female or male participants with black or white skin color. Forty-eight participants equally distributed in terms of sex and skin color participated in the study. Results show an interaction between self-attributes and physical attributes of the observed hand in the right motor cortex of female participants, where corticospinal excitability is increased during observation of hand actions in a different skin color than that of the observer. Our data show that specific physical properties of an observed action modulate motor cortex excitability and we hypothesize that in-group/out-group membership and self-related processes underlie these effects. PMID- 17912351 TI - An expanded set of amino acid analogs for the ribosomal translation of unnatural peptides. AB - BACKGROUND: The application of in vitro translation to the synthesis of unnatural peptides may allow the production of extremely large libraries of highly modified peptides, which are a potential source of lead compounds in the search for new pharmaceutical agents. The specificity of the translation apparatus, however, limits the diversity of unnatural amino acids that can be incorporated into peptides by ribosomal translation. We have previously shown that over 90 unnatural amino acids can be enzymatically loaded onto tRNA. METHODOLOGY/PRINCIPAL FINDINGS: We have now used a competition assay to assess the efficiency of tRNA-aminoacylation of these analogs. We have also used a series of peptide translation assays to measure the efficiency with which these analogs are incorporated into peptides. The translation apparatus tolerates most side chain derivatives, a few alpha,alpha disubstituted, N-methyl and alpha hydroxy derivatives, but no beta-amino acids. We show that over 50 unnatural amino acids can be incorporated into peptides by ribosomal translation. Using a set of analogs that are efficiently charged and translated we were able to prepare individual peptides containing up to 13 different unnatural amino acids. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that a diverse array of unnatural building blocks can be translationally incorporated into peptides. These building blocks provide new opportunities for in vitro selections with highly modified drug-like peptides. PMID- 17912352 TI - The genetic signature of sex-biased migration in patrilocal chimpanzees and humans. AB - A large body of theoretical work suggests that analyses of variation at the maternally inherited mitochondrial (mt)DNA and the paternally inherited non recombining portion of the Y chromosome (NRY) are a potentially powerful way to reveal the differing migratory histories of men and women across human societies. However, the few empirical studies comparing mtDNA and NRY variation and known patterns of sex-biased migration have produced conflicting results. Here we review some methodological reasons for these inconsistencies, and take them into account to provide an unbiased characterization of mtDNA and NRY variation in chimpanzees, one of the few mammalian taxa where males routinely remain in and females typically disperse from their natal groups. We show that patterns of mtDNA and NRY variation are more strongly contrasting in patrilocal chimpanzees compared with patrilocal human societies. The chimpanzee data we present here thus provide a valuable comparative benchmark of the patterns of mtDNA and NRY variation to be expected in a society with extremely female-biased dispersal. PMID- 17912353 TI - Non-invasive in vivo imaging of calcium signaling in mice. AB - Rapid and transient elevations of Ca(2+) within cellular microdomains play a critical role in the regulation of many signal transduction pathways. Described here is a genetic approach for non-invasive detection of localized Ca(2+) concentration ([Ca(2+)]) rises in live animals using bioluminescence imaging (BLI). Transgenic mice conditionally expressing the Ca(2+)-sensitive bioluminescent reporter GFP-aequorin targeted to the mitochondrial matrix were studied in several experimental paradigms. Rapid [Ca(2+)] rises inside the mitochondrial matrix could be readily detected during single-twitch muscle contractions. Whole body patterns of [Ca(2+)] were monitored in freely moving mice and during epileptic seizures. Furthermore, variations in mitochondrial [Ca(2+)] correlated to behavioral components of the sleep/wake cycle were observed during prolonged whole body recordings of newborn mice. This non invasive imaging technique opens new avenues for the analysis of Ca(2+) signaling whenever whole body information in freely moving animals is desired, in particular during behavioral and developmental studies. PMID- 17912354 TI - Membrane recruitment of scaffold proteins drives specific signaling. AB - Cells must give the right response to each stimulus they receive. Scaffolding, a signaling process mediated by scaffold proteins, participates in the decoding of the cues by specifically directing signal transduction. The aim of this paper is to describe the molecular mechanisms of scaffolding, i.e. the principles by which scaffold proteins drive a specific response of the cell. Since similar scaffold proteins are found in many species, they evolved according to the purpose of each organism. This means they require adaptability. In the usual description of the mechanisms of scaffolding, scaffold proteins are considered as reactors where molecules involved in a cascade of reactions are simultaneously bound with the right orientation to meet and interact. This description is not realistic: (i) it is not verified by experiments and (ii) timing and orientation constraints make it complex which seems to contradict the required adaptability. A scaffold protein, Ste5, is used in the MAPK pathway of Saccharomyces cerevisiae for the cell to provide a specific response to stimuli. The massive amount of data available for this pathway makes it ideal to investigate the actual mechanisms of scaffolding. Here, a complete treatment of the chemical reactions allows the computation of the distributions of all the proteins involved in the MAPK pathway when the cell receives various cues. These distributions are compared to several experimental results. It turns out that the molecular mechanisms of scaffolding are much simpler and more adaptable than previously thought in the reactor model. Scaffold proteins bind only one molecule at a time. Then, their membrane recruitment automatically drives specific, amplified and localized signal transductions. The mechanisms presented here, which explain how the membrane recruitment of a protein can produce a drastic change in the activity of cells, are generic and may be commonly used in many biological processes. PMID- 17912355 TI - Haemoglobin C and S role in acquired immunity against Plasmodium falciparum malaria. AB - A recently proposed mechanism of protection for haemoglobin C (HbC; beta6Glu- >Lys) links an abnormal display of PfEMP1, an antigen involved in malaria pathogenesis, on the surface of HbC infected erythrocytes together with the observation of reduced cytoadhesion of parasitized erythrocytes and impaired rosetting in vitro. We investigated the impact of this hypothesis on the development of acquired immunity against Plasmodium falciparum variant surface antigens (VSA) encoding PfEMP1 in HbC in comparison with HbA and HbS carriers of Burkina Faso. We measured: i) total IgG against a single VSA, A4U, and against a panel of VSA from severe malaria cases in human sera from urban and rural areas of Burkina Faso of different haemoglobin genotypes (CC, AC, AS, SC, SS); ii) total IgG against recombinant proteins of P. falciparum asexual sporozoite, blood stage antigens, and parasite schizont extract; iii) total IgG against tetanus toxoid. Results showed that the reported abnormal cell-surface display of PfEMP1 on HbC infected erythrocytes observed in vitro is not associated to lower anti- PfEMP1 response in vivo. Higher immune response against the VSA panel and malaria antigens were observed in all adaptive genotypes containing at least one allelic variant HbC or HbS in the low transmission urban area whereas no differences were detected in the high transmission rural area. In both contexts the response against tetanus toxoid was not influenced by the beta-globin genotype. These findings suggest that both HbC and HbS affect the early development of naturally acquired immunity against malaria. The enhanced immune reactivity in both HbC and HbS carriers supports the hypothesis that the protection against malaria of these adaptive genotypes might be at least partially mediated by acquired immunity against malaria. PMID- 17912356 TI - Ewing sarcoma protein ewsr1 maintains mitotic integrity and proneural cell survival in the zebrafish embryo. AB - BACKGROUND: The Ewing sarcoma breakpoint region 1 gene (EWSR1), also known as EWS, is fused to a number of different partner genes as a result of chromosomal translocation in diverse sarcomas. Despite the involvement of EWSR1 in these diverse sarcomas, the in vivo function of wild type EWSR1 remains unclear. PRINCIPAL FINDINGS: We identified two zebrafish EWSR1 orthologues, ewsr1a and ewsr1b, and demonstrate that both genes are expressed maternally, and are expressed ubiquitously throughout zebrafish embryonic development. Morpholino induced knockdown of both zebrafish ewsr1 genes led to mitotic defects with multipolar or otherwise abnormal mitotic spindles starting from the bud stage (10 hour post-fertilization (hpf)). The abnormalities in mitotic spindles were followed by p53-mediated apoptosis in the developing central nervous system (CNS) leading to a reduction in the number of proneural cells, disorganization of neuronal networks, and embryonic lethality by 5 days post-fertilization. siRNA silencing of EWSR1 in Hela cells resulted in mitotic defects accompanied by apoptotic cell death, indicating that the role of EWSR1 is conserved between zebrafish and human. CONCLUSIONS: Ewsr1 maintains mitotic integrity and proneural cell survival in early zebrafish development. PMID- 17912357 TI - Effects of insemination quantity on honey bee queen physiology. AB - Mating has profound effects on the physiology and behavior of female insects, and in honey bee (Apis mellifera) queens, these changes are permanent. Queens mate with multiple males during a brief period in their early adult lives, and shortly thereafter they initiate egg-laying. Furthermore, the pheromone profiles of mated queens differ from those of virgins, and these pheromones regulate many different aspects of worker behavior and colony organization. While it is clear that mating causes dramatic changes in queens, it is unclear if mating number has more subtle effects on queen physiology or queen-worker interactions; indeed, the effect of multiple matings on female insect physiology has not been broadly addressed. Because it is not possible to control the natural mating behavior of queens, we used instrumental insemination and compared queens inseminated with semen from either a single drone (single-drone inseminated, or SDI) or 10 drones (multi drone inseminated, or MDI). We used observation hives to monitor attraction of workers to SDI or MDI queens in colonies, and cage studies to monitor the attraction of workers to virgin, SDI, and MDI queen mandibular gland extracts (the main source of queen pheromone). The chemical profiles of the mandibular glands of virgin, SDI, and MDI queens were characterized using GC-MS. Finally, we measured brain expression levels in SDI and MDI queens of a gene associated with phototaxis in worker honey bees (Amfor). Here, we demonstrate for the first time that insemination quantity significantly affects mandibular gland chemical profiles, queen-worker interactions, and brain gene expression. Further research will be necessary to elucidate the mechanistic bases for these effects: insemination volume, sperm and seminal protein quantity, and genetic diversity of the sperm may all be important factors contributing to this profound change in honey bee queen physiology, queen behavior, and social interactions in the colony. PMID- 17912358 TI - Germ line origin and somatic mutations determine the target tissues in systemic AL-amyloidosis. AB - BACKGROUND: Amyloid is insoluble aggregated proteins deposited in the extra cellular space. About 25 different proteins are known to form amyloid in vivo and are associated with severe diseases such as Alzheimer's disease, prion diseases and type-2 diabetes. Light chain (AL) -amyloidosis is unique among amyloid diseases in that the fibril protein, a monoclonal immunoglobulin light chain, varies between individuals and that no two AL-proteins with identical primary structures have been described to date. The variability in tissue distribution of amyloid deposits is considerably larger in systemic AL-amyloidosis than in any other form of amyloidosis. The reason for this variation is believed to be based on the differences in properties of the amyloidogenic immunoglobulin light chain. However, there is presently no known relationship between the structure of an AL protein and tissue distribution. METHODOLOGY/PRINCIPAL FINDINGS: We compared the pattern of amyloid deposition in four individuals with amyloid protein derived from variable light chain gene O18-O8, the source of a high proportion of amyloidogenic light chains, and in whom all or most of the fibril protein had been determined by amino acid sequencing. In spite of great similarities between the structures of the proteins, there was a pronounced variability in deposition pattern. We also compared the tissue distribution in these four individuals with that of four other patients with AL-amyloid derived from the L2-L16 gene. Although the interindividual variations were pronounced, liver and kidney involvement was much more evident in the latter four. CONCLUSIONS/SIGNIFICANCE: We conclude that although the use of a specific gene influences the tissue distribution of amyloid, each light chain exhibits one or more determinants of organ-specificity, which originate from somatic mutations and post-translational modifications. Eventual identification of such determinants could lead to improved treatment of patients with AL amyloidosis. PMID- 17912359 TI - Conservation, variability and the modeling of active protein kinases. AB - The human proteome is rich with protein kinases, and this richness has made the kinase of crucial importance in initiating and maintaining cell behavior. Elucidating cell signaling networks and manipulating their components to understand and alter behavior require well designed inhibitors. These inhibitors are needed in culture to cause and study network perturbations, and the same compounds can be used as drugs to treat disease. Understanding the structural biology of protein kinases in detail, including their commonalities, differences and modes of substrate interaction, is necessary for designing high quality inhibitors that will be of true use for cell biology and disease therapy. To this end, we here report on a structural analysis of all available active-conformation protein kinases, discussing residue conservation, the novel features of such conservation, unique properties of atypical kinases and variability in the context of substrate binding. We also demonstrate how this information can be used for structure prediction. Our findings will be of use not only in understanding protein kinase function and evolution, but they highlight the flaws inherent in kinase drug design as commonly practiced and dictate an appropriate strategy for the sophisticated design of specific inhibitors for use in the laboratory and disease therapy. PMID- 17912360 TI - Investigating the role of islet cytoarchitecture in its oscillation using a new beta-cell cluster model. AB - The oscillatory insulin release is fundamental to normal glycemic control. The basis of the oscillation is the intercellular coupling and bursting synchronization of beta cells in each islet. The functional role of islet beta cell mass organization with respect to its oscillatory bursting is not well understood. This is of special interest in view of the recent finding of islet cytoarchitectural differences between human and animal models. In this study we developed a new hexagonal closest packing (HCP) cell cluster model. The model captures more accurately the real islet cell organization than the simple cubic packing (SCP) cluster that is conventionally used. Using our new model we investigated the functional characteristics of beta-cell clusters, including the fraction of cells able to burst f(b), the synchronization index lambda of the bursting beta cells, the bursting period T(b), the plateau fraction p(f), and the amplitude of intracellular calcium oscillation [Ca]. We determined their dependence on cluster architectural parameters including number of cells n(beta), number of inter-beta cell couplings of each beta cell n(c), and the coupling strength g(c). We found that at low values of n(beta), n(c) and g(c), the oscillation regularity improves with their increasing values. This functional gain plateaus around their physiological values in real islets, at n(beta) approximately 100, n(c) approximately 6 and g(c) approximately 200 pS. In addition, normal beta-cell clusters are robust against significant perturbation to their architecture, including the presence of non-beta cells or dead beta cells. In clusters with n(beta)> approximately 100, coordinated beta-cell bursting can be maintained at up to 70% of beta-cell loss, which is consistent with laboratory and clinical findings of islets. Our results suggest that the bursting characteristics of a beta-cell cluster depend quantitatively on its architecture in a non-linear fashion. These findings are important to understand the islet bursting phenomenon and the regulation of insulin secretion, under both physiological and pathological conditions. PMID- 17912361 TI - Design and pre-clinical evaluation of a universal HIV-1 vaccine. AB - BACKGROUND: One of the big roadblocks in development of HIV-1/AIDS vaccines is the enormous diversity of HIV-1, which could limit the value of any HIV-1 vaccine candidate currently under test. METHODOLOGY AND FINDINGS: To address the HIV-1 variation, we designed a novel T cell immunogen, designated HIV(CONSV), by assembling the 14 most conserved regions of the HIV-1 proteome into one chimaeric protein. Each segment is a consensus sequence from one of the four major HIV-1 clades A, B, C and D, which alternate to ensure equal clade coverage. The gene coding for the HIV(CONSV) protein was inserted into the three most studied vaccine vectors, plasmid DNA, human adenovirus serotype 5 and modified vaccine virus Ankara (MVA), and induced HIV-1-specific T cell responses in mice. We also demonstrated that these conserved regions prime CD8(+) and CD4(+) T cell to highly conserved epitopes in humans and that these epitopes, although usually subdominant, generate memory T cells in patients during natural HIV-1 infection. SIGNIFICANCE: Therefore, this vaccine approach provides an attractive and testable alternative for overcoming the HIV-1 variability, while focusing T cell responses on regions of the virus that are less likely to mutate and escape. Furthermore, this approach has merit in the simplicity of design and delivery, requiring only a single immunogen to provide extensive coverage of global HIV-1 population diversity. PMID- 17912362 TI - A multidirectional non-cell autonomous control and a genetic interaction restricting tobacco etch virus susceptibility in Arabidopsis. AB - BACKGROUND: Viruses constitute a major class of pathogens that infect a variety of hosts. Understanding the intricacies of signaling during host-virus interactions should aid in designing disease prevention strategies and in understanding mechanistic aspects of host and pathogen signaling machinery. METHODOLOGY/PRINCIPAL FINDINGS: An Arabidopsis mutant, B149, impaired in susceptibility to Tobacco etch virus (TEV), a positive strand RNA virus of picoRNA family, was identified using a high-throughput genetic screen and a counterselection scheme. The defects include initiation of infection foci, rate of cell-to-cell movement and long distance movement. CONCLUSIONS/SIGNIFICANCE: The defect in infectivity is conferred by a recessive locus. Molecular genetic analysis and complementation analysis with three alleles of a previously published mutant lsp1 (loss of susceptibility to potyviruses) indicate a genetic interaction conferring haploinsufficiency between the B149 locus and certain alleles of lsp1 resulting in impaired host susceptibility. The pattern of restriction of TEV foci on leaves at or near the boundaries of certain cell types and leaf boundaries suggest dysregulation of a multidirectional non-cell autonomous regulatory mechanism. Understanding the nature of this multidirectional signal and the molecular genetic mechanism conferring it should potentially reveal a novel arsenal in the cellular machinery. PMID- 17912363 TI - Antiviral oseltamivir is not removed or degraded in normal sewage water treatment: implications for development of resistance by influenza A virus. AB - Oseltamivir is the main antiviral for treatment and prevention of pandemic influenza. The increase in oseltamivir resistance reported recently has therefore sparked a debate on how to use oseltamivir in non pandemic influenza and the risks associated with wide spread use during a pandemic. Several questions have been asked about the fate of oseltamivir in the sewage treatment plants and in the environment. We have assessed the fate of oseltamivir and discuss the implications of environmental residues of oseltamivir regarding the occurrence of resistance. A series of batch experiments that simulated normal sewage treatment with oseltamivir present was conducted and the UV-spectra of oseltamivir were recorded. FINDINGS: Our experiments show that the active moiety of oseltamivir is not removed in normal sewage water treatments and is not degraded substantially by UV light radiation, and that the active substance is released in waste water leaving the plant. Our conclusion is that a ubiquitous use of oseltamivir may result in selection pressures in the environment that favor development of drug resistance. PMID- 17912364 TI - Carbon dioxide inhalation induces dose-dependent and age-related negative affectivity. AB - BACKGROUND: Carbon dioxide inhalation is known to induce an emotion similar to spontaneous panic in Panic Disorder patients. The affective response to carbon dioxide in healthy subjects was not clearly characterized yet. METHODOLOGY/PRINCIPAL FINDINGS: Sixty-four healthy subjects underwent a double inhalation of four mixtures containing respectively 0, 9, 17.5 and 35% CO(2) in compressed air, following a double blind, cross-over, randomized design. Affective responses were assessed according to DSM IV criteria for panic, using an Electronic Visual Analogue Scale and the Panic Symptom List. It was demonstrated that carbon dioxide challenges induced a dose dependent negative affect (p<0.0001). This affect was semantically identical to the DSM IV definition of panic. Older individuals were subjectively less sensitive to Carbon Dioxide (p<0.05). CONCLUSIONS/SIGNIFICANCE: CO(2) induced affectivity may lay on a continuum with pathological panic attacks. Consistent with earlier suggestions that panic is a false biological alarm, the affective response to CO(2) may be part of a protective system triggered by suffocation and acute metabolic distress. PMID- 17912365 TI - An improved, bias-reduced probabilistic functional gene network of baker's yeast, Saccharomyces cerevisiae. AB - BACKGROUND: Probabilistic functional gene networks are powerful theoretical frameworks for integrating heterogeneous functional genomics and proteomics data into objective models of cellular systems. Such networks provide syntheses of millions of discrete experimental observations, spanning DNA microarray experiments, physical protein interactions, genetic interactions, and comparative genomics; the resulting networks can then be easily applied to generate testable hypotheses regarding specific gene functions and associations. METHODOLOGY/PRINCIPAL FINDINGS: We report a significantly improved version (v. 2) of a probabilistic functional gene network of the baker's yeast, Saccharomyces cerevisiae. We describe our optimization methods and illustrate their effects in three major areas: the reduction of functional bias in network training reference sets, the application of a probabilistic model for calculating confidences in pair-wise protein physical or genetic interactions, and the introduction of simple thresholds that eliminate many false positive mRNA co-expression relationships. Using the network, we predict and experimentally verify the function of the yeast RNA binding protein Puf6 in 60S ribosomal subunit biogenesis. CONCLUSIONS/SIGNIFICANCE: YeastNet v. 2, constructed using these optimizations together with additional data, shows significant reduction in bias and improvements in precision and recall, in total covering 102,803 linkages among 5,483 yeast proteins (95% of the validated proteome). YeastNet is available from http://www.yeastnet.org. PMID- 17912366 TI - Deletion of genes implicated in protecting the integrity of male germ cells has differential effects on the incidence of DNA breaks and germ cell loss. AB - BACKGROUND: Infertility affects approximately 20% of couples in Europe and in 50% of cases the problem lies with the male partner. The impact of damaged DNA originating in the male germ line on infertility is poorly understood but may increase miscarriage. Mouse models allow us to investigate how deficiencies in DNA repair/damage response pathways impact on formation and function of male germ cells. We have investigated mice with deletions of ERCC1 (excision repair cross complementing gene 1), MSH2 (MutS homolog 2, involved in mismatch repair pathway), and p53 (tumour suppressor gene implicated in elimination of germ cells with DNA damage). PRINCIPAL FINDINGS: We demonstrate for the first time that depletion of ERCC1 or p53 from germ cells results in an increased incidence of unrepaired DNA breaks in pachytene spermatocytes and increased numbers of caspase 3 positive (apoptotic) germ cells. Sertoli cell-only tubules were detected in testes from mice lacking expression of ERCC1 or MSH2 but not p53. The number of sperm recovered from epididymes was significantly reduced in mice lacking testicular ERCC1 and 40% of sperm contained DNA breaks whereas the numbers of sperm were not different to controls in adult Msh2 -/- or p53 -/- mice nor did they have significantly compromised DNA. CONCLUSIONS: These data have demonstrated that deletion of Ercc1, Msh2 and p53 can have differential but overlapping affects on germ cell function and sperm production. These findings increase our understanding of the ways in which gene mutations can have an impact on male fertility. PMID- 17912367 TI - Effect of cellular quiescence on the success of targeted CML therapy. AB - BACKGROUND: Similar to tissue stem cells, primitive tumor cells in chronic myelogenous leukemia have been observed to undergo quiescence; that is, the cells can temporarily stop dividing. Using mathematical models, we investigate the effect of cellular quiescence on the outcome of therapy with targeted small molecule inhibitors. METHODS AND RESULTS: According to the models, the initiation of treatment can result in different patterns of tumor cell decline: a biphasic decline, a one-phase decline, and a reverse biphasic decline. A biphasic decline involves a fast initial phase (which roughly corresponds to the eradication of cycling cells by the drug), followed by a second and slower phase of exponential decline (corresponding to awakening and death of quiescent cells), which helps explain clinical data. We define the time when the switch to the second phase occurs, and identify parameters that determine whether therapy can drive the tumor extinct in a reasonable period of time or not. We further ask how cellular quiescence affects the evolution of drug resistance. We find that it has no effect on the probability that resistant mutants exist before therapy if treatment occurs with a single drug, but that quiescence increases the probability of having resistant mutants if patients are treated with a combination of two or more drugs with different targets. Interestingly, while quiescence prolongs the time until therapy reduces the number of cells to low levels or extinction, the therapy phase is irrelevant for the evolution of drug resistant mutants. If treatment fails as a result of resistance, the mutants will have evolved during the tumor growth phase, before the start of therapy. Thus, prevention of resistance is not promoted by reducing the quiescent cell population during therapy (e.g., by a combination of cell activation and drug mediated killing). CONCLUSIONS: The mathematical models provide insights into the effect of quiescence on the basic kinetics of the response to targeted treatment of CML. They identify determinants of success in the absence of drug resistant mutants, and elucidate how quiescence influences the emergence of drug resistant mutants. PMID- 17912369 TI - Deletion of PKBalpha/Akt1 affects thymic development. AB - BACKGROUND: The thymus constitutes the primary lymphoid organ for the majority of T cells. The phosphatidyl-inositol 3 kinase (PI3K) signaling pathway is involved in lymphoid development. Defects in single components of this pathway prevent thymocytes from progressing beyond early T cell developmental stages. Protein kinase B (PKB) is the main effector of the PI3K pathway. METHODOLOGY/PRINCIPAL FINDINGS: To determine whether PKB mediates PI3K signaling in the thymus, we characterized PKB knockout thymi. Our results reveal a significant thymic hypocellularity in PKBalpha(-/-) neonates and an accumulation of early thymocyte subsets in PKBalpha(-/-) adult mice. Using thymic grafting and fetal liver cell transfer experiments, the latter finding was specifically attributed to the lack of PKBalpha within the lymphoid component of the thymus. Microarray analyses show that the absence of PKBalpha in early thymocyte subsets modifies the expression of genes known to be involved in pre-TCR signaling, in T cell activation, and in the transduction of interferon-mediated signals. CONCLUSIONS/SIGNIFICANCE: This report highlights the specific requirements of PKBalpha for thymic development and opens up new prospects as to the mechanism downstream of PKBalpha in early thymocytes. PMID- 17912368 TI - Creatine monohydrate and conjugated linoleic acid improve strength and body composition following resistance exercise in older adults. AB - Aging is associated with lower muscle mass and an increase in body fat. We examined whether creatine monohydrate (CrM) and conjugated linoleic acid (CLA) could enhance strength gains and improve body composition (i.e., increase fat free mass (FFM); decrease body fat) following resistance exercise training in older adults (>65 y). Men (N = 19) and women (N = 20) completed six months of resistance exercise training with CrM (5g/d)+CLA (6g/d) or placebo with randomized, double blind, allocation. Outcomes included: strength and muscular endurance, functional tasks, body composition (DEXA scan), blood tests (lipids, liver function, CK, glucose, systemic inflammation markers (IL-6, C-reactive protein)), urinary markers of compliance (creatine/creatinine), oxidative stress (8-OH-2dG, 8-isoP) and bone resorption (Nu-telopeptides). Exercise training improved all measurements of functional capacity (P<0.05) and strength (P<0.001), with greater improvement for the CrM+CLA group in most measurements of muscular endurance, isokinetic knee extension strength, FFM, and lower fat mass (P<0.05). Plasma creatinine (P<0.05), but not creatinine clearance, increased for CrM+CLA, with no changes in serum CK activity or liver function tests. Together, this data confirms that supervised resistance exercise training is safe and effective for increasing strength in older adults and that a combination of CrM and CLA can enhance some of the beneficial effects of training over a six-month period. Trial Registration. ClinicalTrials.gov NCT00473902. PMID- 17912370 TI - Structure and reaction mechanism of basil eugenol synthase. AB - Phenylpropenes, a large group of plant volatile compounds that serve in multiple roles in defense and pollinator attraction, contain a propenyl side chain. Eugenol synthase (EGS) catalyzes the reductive displacement of acetate from the propenyl side chain of the substrate coniferyl acetate to produce the allyl phenylpropene eugenol. We report here the structure determination of EGS from basil (Ocimum basilicum) by protein x-ray crystallography. EGS is structurally related to the short-chain dehydrogenase/reductases (SDRs), and in particular, enzymes in the isoflavone-reductase-like subfamily. The structure of a ternary complex of EGS bound to the cofactor NADP(H) and a mixed competitive inhibitor EMDF ((7S,8S)-ethyl (7,8-methylene)-dihydroferulate) provides a detailed view of the binding interactions within the EGS active site and a starting point for mutagenic examination of the unusual reductive mechanism of EGS. The key interactions between EMDF and the EGS-holoenzyme include stacking of the phenyl ring of EMDF against the cofactor's nicotinamide ring and a water-mediated hydrogen-bonding interaction between the EMDF 4-hydroxy group and the side-chain amino moiety of a conserved lysine residue, Lys132. The C4 carbon of nicotinamide resides immediately adjacent to the site of hydride addition, the C7 carbon of cinnamyl acetate substrates. The inhibitor-bound EGS structure suggests a two step reaction mechanism involving the formation of a quinone-methide prior to reduction. The formation of this intermediate is promoted by a hydrogen-bonding network that favors deprotonation of the substrate's 4-hydroxyl group and disfavors binding of the acetate moiety, akin to a push-pull catalytic mechanism. Notably, the catalytic involvement in EGS of the conserved Lys132 in preparing the phenolic substrate for quinone methide formation through the proton-relay network appears to be an adaptation of the analogous role in hydrogen bonding played by the equivalent lysine residue in other enzymes of the SDR family. PMID- 17912372 TI - Phylogeny, diet, and cranial integration in australodelphian marsupials. AB - Studies of morphological integration provide valuable information on the correlated evolution of traits and its relationship to long-term patterns of morphological evolution. Thus far, studies of morphological integration in mammals have focused on placentals and have demonstrated that similarity in integration is broadly correlated with phylogenetic distance and dietary similarity. Detailed studies have also demonstrated a significant correlation between developmental relationships among structures and adult morphological integration. However, these studies have not yet been applied to marsupial taxa, which differ greatly from placentals in reproductive strategy and cranial development and could provide the diversity necessary to assess the relationships among phylogeny, ecology, development, and cranial integration. This study presents analyses of morphological integration in 20 species of australodelphian marsupials, and shows that phylogeny is significantly correlated with similarity of morphological integration in most clades. Size-related correlations have a significant affect on results, particularly in Peramelia, which shows a striking decrease in similarity of integration among species when size is removed. Diet is not significantly correlated with similarity of integration in any marsupial clade. These results show that marsupials differ markedly from placental mammals in the relationships of cranial integration, phylogeny, and diet, which may be related to the accelerated development of the masticatory apparatus in marsupials. PMID- 17912371 TI - Ant species differences determined by epistasis between brood and worker genomes. AB - Epistasis arising from physiological interactions between gene products often contributes to species differences, particularly those involved in reproductive isolation. In social organisms, phenotypes are influenced by the genotypes of multiple interacting individuals. In theory, social interactions can give rise to an additional type of epistasis between the genomes of social partners that can contribute to species differences. Using a full-factorial cross-fostering design with three species of closely related Temnothorax ants, I found that adult worker size was determined by an interaction between the genotypes of developing brood and care-giving workers, i.e. intergenomic epistasis. Such intergenomic social epistasis provides a strong signature of coevolution between social partners. These results demonstrate that just as physiologically interacting genes coevolve, diverge, and contribute to species differences, so do socially interacting genes. Coevolution and conflict between social partners, especially relatives such as parents and offspring, has long been recognized as having widespread evolutionary effects. This coevolutionary process may often result in coevolved socially-interacting gene complexes that contribute to species differences. PMID- 17912373 TI - A genetic code alteration is a phenotype diversity generator in the human pathogen Candida albicans. AB - BACKGROUND: The discovery of genetic code alterations and expansions in both prokaryotes and eukaryotes abolished the hypothesis of a frozen and universal genetic code and exposed unanticipated flexibility in codon and amino acid assignments. It is now clear that codon identity alterations involve sense and non-sense codons and can occur in organisms with complex genomes and proteomes. However, the biological functions, the molecular mechanisms of evolution and the diversity of genetic code alterations remain largely unknown. In various species of the genus Candida, the leucine CUG codon is decoded as serine by a unique serine tRNA that contains a leucine 5'-CAG-3'anticodon (tRNA(CAG)(Ser)). We are using this codon identity redefinition as a model system to elucidate the evolution of genetic code alterations. METHODOLOGY/PRINCIPAL FINDINGS: We have reconstructed the early stages of the Candida genetic code alteration by engineering tRNAs that partially reverted the identity of serine CUG codons back to their standard leucine meaning. Such genetic code manipulation had profound cellular consequences as it exposed important morphological variation, altered gene expression, re-arranged the karyotype, increased cell-cell adhesion and secretion of hydrolytic enzymes. CONCLUSION/SIGNIFICANCE: Our study provides the first experimental evidence for an important role of genetic code alterations as generators of phenotypic diversity of high selective potential and supports the hypothesis that they speed up evolution of new phenotypes. PMID- 17912374 TI - Siglecg limits the size of B1a B cell lineage by down-regulating NFkappaB activation. AB - BACKGROUND: B1 B cells are believed to be a unique lineage with a distinct developmental pathway, function and activation requirement. How this lineage is genetically determined remained largely obscure. METHODS AND PRINCIPAL FINDINGS: Using the Siglecg-deficient mice with a knockin of green-fluorescent protein encoding sequence, we show here that, although the Siglecg gene is broadly expressed at high levels in all stages and/or lineages of B cells tested and at lower levels in other lineages, its deletion selectively expanded the B1a B cell lineages, including the frequency of the B1 cell progenitor in the bone marrow and the number of B1a cells in the peritoneal cavity, by postnatal expansion. The expansion of B1a B cells in the peritoneal correlated with enhanced activation of NFkappaB and was ablated by an IKK inhibitor. CONCLUSION AND SIGNIFICANCE: Our data revealed a critical role for Siglec G-NFkappaB pathway in regulating B1a B cell lineage. These data lead to a novel model of B1a lineage development that explains a large array of genetic data in this field. PMID- 17912376 TI - Inflated impact factors? The true impact of evolutionary papers in non evolutionary journals. AB - Amongst the numerous problems associated with the use of impact factors as a measure of quality are the systematic differences in impact factors that exist among scientific fields. While in theory this can be circumvented by limiting comparisons to journals within the same field, for a diverse and multidisciplinary field like evolutionary biology, in which the majority of papers are published in journals that publish both evolutionary and non evolutionary papers, this is impossible. However, a journal's overall impact factor may well be a poor predictor for the impact of its evolutionary papers. The extremely high impact factors of some multidisciplinary journals, for example, are by many believed to be driven mostly by publications from other fields. Despite plenty of speculation, however, we know as yet very little about the true impact of evolutionary papers in journals not specifically classified as evolutionary. Here I present, for a wide range of journals, an analysis of the number of evolutionary papers they publish and their average impact. I show that there are large differences in impact among evolutionary and non-evolutionary papers within journals; while the impact of evolutionary papers published in multidisciplinary journals is substantially overestimated by their overall impact factor, the impact of evolutionary papers in many of the more specialized, non evolutionary journals is significantly underestimated. This suggests that, for evolutionary biologists, publishing in high-impact multidisciplinary journals should not receive as much weight as it does now, while evolutionary papers in more narrowly defined journals are currently undervalued. Importantly, however, their ranking remains largely unaffected. While journal impact factors may thus indeed provide a meaningful qualitative measure of impact, a fair quantitative comparison requires a more sophisticated journal classification system, together with multiple field-specific impact statistics per journal. PMID- 17912377 TI - Molecularly imprinted polymers (MIPs): sensing, an explosive new opportunity? AB - Our group is currently developing in-field detection systems alongside the Australian Federal Police Forensic Services utilising molecularly imprinted polymers as the recognition elements. This review looks at MIP synthesis and our perceptions of future directions from an Australian and forensic perspective. PMID- 17912375 TI - Composition and hierarchical organisation of a spider silk. AB - Albeit silks are fairly well understood on a molecular level, their hierarchical organisation and the full complexity of constituents in the spun fibre remain poorly defined. Here we link morphological defined structural elements in dragline silk of Nephila clavipes to their biochemical composition and physicochemical properties. Five layers of different make-ups could be distinguished. Of these only the two core layers contained the known silk proteins, but all can vitally contribute to the mechanical performance or properties of the silk fibre. Understanding the composite nature of silk and its supra-molecular organisation will open avenues in the production of high performance fibres based on artificially spun silk material. PMID- 17912378 TI - Total synthesis approaches to natural product derivatives based on the combination of chemical synthesis and metabolic engineering. AB - Secondary metabolites are an extremely diverse and important group of natural products with industrial and biomedical implications. Advances in metabolic engineering of both native and heterologous secondary metabolite producing organisms have allowed the directed synthesis of desired novel products by exploiting their biosynthetic potentials. Metabolic engineering utilises knowledge of cellular metabolism to alter biosynthetic pathways. An important technique that combines chemical synthesis with metabolic engineering is mutasynthesis (mutational biosynthesis; MBS), which advanced from precursor directed biosynthesis (PDB). Both techniques are based on the cellular uptake of modified biosynthetic intermediates and their incorporation into complex secondary metabolites. Mutasynthesis utilises genetically engineered organisms in conjunction with feeding of chemically modified intermediates. From a synthetic chemist's point of view the concept of mutasynthesis is highly attractive, as the method combines chemical expertise with Nature's synthetic machinery and thus can be exploited to rapidly create small libraries of secondary metabolites. However, in each case, the method has to be critically compared with semi- and total synthesis in terms of practicability and efficiency. Recent developments in metabolic engineering promise to further broaden the scope of outsourcing chemically demanding steps to biological systems. PMID- 17912379 TI - Recent highlights in modified oligonucleotide chemistry. AB - The synthesis of modified nucleic acids has been the subject of much study ever since the structure of DNA was elucidated by Watson and Crick at Cambridge and Wilkins and Franklin at King's College over half a century ago. This review describes recent developments in the synthesis and application of these artificial nucleic acids, predominantly the phosphoramidites which allow for easy inclusion into oligonucleotides, and is divided into three separate sections. Firstly, modifications to the base portion will be discussed followed secondly by modifications to the sugar portion. Finally, changes in the type of nucleic acid linker will be discussed in the third section. Peptide Nucleic Acids (PNAs) are not discussed in this review as they represent a separate and large area of nucleic acid mimics. PMID- 17912380 TI - Refined models for the preferential interactions of tryptophan with phosphocholines. AB - A series of molecular models of the adducts formed between N-acetyl-l-tryptophan ethylamide and diacetyl-sn-glycero-3-phosphocholine have been generated. Using rOesy data that enabled us to place restrictions on the proximity of a number of key protons in the amino acid/phosphocholine pairs, a series of structures were generated following molecular dynamics and mechanics experiments using the CHARMM27 force field. These structures were then subjected to a series of clustering algorithms in order to classify the tight binding interactions between a single tryptophan and a phosphocholine. From these analyses, it is evident that: (i) binding is characterised by hydrogen bonding between the indole NH as donor and phosphate oxygen as acceptor, cation-carbonyl interactions between the choline ammonium and amide carbonyl groups and cation-pi interactions; (ii) cation-pi interactions are not always observed, particularly when their formation is at the expense of cation-carbonyl and hydrogen bonding interactions; (iii) on the basis of amino acid torsional parameters, it is possible to predict whether the phosphocholine headgroup will bind in a compact or elongated conformation. Extension of the procedures to characterise 2 : 1 Trp-PC binding revealed that the same intermolecular interactions are predominant; however, combinations of all three intermolecular interactions within the same adduct occur much more frequently due to the availability of donor/acceptor groups from both tryptophans in the 2 : 1 system. PMID- 17912381 TI - Competitive inhibition of aristolochene synthase by phenyl-substituted farnesyl diphosphates: evidence of active site plasticity. AB - Analogues of farnesyl diphosphate (FPP, ) containing phenyl substituents in place of methyl groups have been prepared in syntheses that feature use of a Suzuki Miyaura reaction as a key step. These analogues were found not to act as substrates of the sesquiterpene cyclase aristolochene synthase from Penicillium roqueforti (AS). However, they were potent competitive inhibitors of AS with K(I) values ranging from 0.8 to 1.2 microM. These results indicate that the diphosphate group contributes the largest part to the binding of the substrate to AS and that the active sites of terpene synthases are sufficiently flexible to accommodate even substrate analogues with large substituents suggesting a potential way for the generation of non-natural terpenoids. Molecular mechanics simulations of the enzyme bound inhibitors suggested that small changes in orientations of active site residues and subtle alterations of the conformation of the backbones of the inhibitors are sufficient to accommodate the phenyl farnesyl-diphosphates. PMID- 17912382 TI - Synthesis and biological evaluation of potential bisubstrate inhibitors of protein farnesyltransferase. Design and synthesis of functionalized imidazoles. AB - A novel series of compounds, derived from 2,5-functionalized imidazoles, have been synthesized as potential bisubstrate inhibitors of protein farnesyltransferase (FTase) using structure-based design. These compounds have a 1,4-diacid chain and a tripeptide connected by an imidazole ring. The synthetic strategy relies on the functionalization at the C-2 position of the heterocycle with the diacid side chain and peptide coupling at the C-5 position. Several new compounds were synthesized in good yields. Kinetic experiments on the most active compounds revealed different binding modes depending on the diacid chain length. PMID- 17912383 TI - Identification of a metagenome-derived esterase with high enantioselectivity in the kinetic resolution of arylaliphatic tertiary alcohols. AB - 35 metagenome-derived esterases bearing a GGG(A)X motif were screened for activity and enantioselectivity in the hydrolysis of a range of tertiary alcohol acetates. Most of the active esterases showed little or no enantioselectivity in the hydrolysis of the terpinyl acetate, linalyl acetate and 3-methylpent-1-yn-3 yl acetate. However, one esterase showed excellent enantioselectivity (E > 100) in the kinetic resolution of 1,1,1-trifluoro-2-phenylbut-3-yn-2-yl acetate as confirmed by a preparative scale reaction. PMID- 17912385 TI - An ab initio and DFT study of some halogen atom transfer reactions from alkyl groups to acyl radical. AB - Ab initio calculations using the 6-311G**, cc-pVDZ, and (valence) double-zeta pseudopotential (DZP) basis sets, with (MP2, QCISD, CCSD(T)) and without (HF) the inclusion of electron correlation, and density functional (BHandHLYP, B3LYP) calculations predict that the transition states for the reaction of acetyl radical with several alkyl halides adopt an almost collinear arrangement of attacking and leaving radicals at the halogen atom. Energy barriers (DeltaE(double dagger)) for these halogen transfer reactions of between 89.2 (chlorine transfer from methyl group) and 25.3 kJ mol(-1) (iodine transfer from tert-butyl group) are calculated at the BHandHLYP/DZP level of theory. While the difference in forward and reverse energy barriers for iodine transfer to acetyl radical is predicted to be 15.1 kJ mol(-1) for primary alkyl iodide, these values are calculated to be 6.7 and -4.2 kJ mol(-1) for secondary and tertiary alkyl iodide respectively. These data are in good agreement with available experimental data in that atom transfer radical carbonylation reactions are sluggish with primary alkyl iodides, but proceed smoothly with secondary and tertiary alkyl iodides. These calculations also predict that bromine transfer reactions involving acyl radical are also feasible at moderately high temperature. PMID- 17912384 TI - Synthesis of 6-formylpterin nucleoside analogs and their ROS generation activities in the presence of NADH in the dark. AB - We demonstrated previously that 3-position-modified 6-formylpterin (6FP) derivatives produce reactive oxygen species (ROS) such as hydrogen peroxide (H(2)O(2)) from oxygen in the presence of NADH in the dark. It has been shown that 6FP derivatives markedly generate ROS, which gives rise to their particular physiological activities, such as induction of apoptosis in cellular and living systems, suggesting that such compounds provide a hint for the design of a ROS controlling agent in vivo. However, it is not well understood why such unique activities appear on chemical modification. In the present study, in order to see the effect on ROS generation activity in the dark by the modification of the 1 position in 6FP, we have developed a new synthetic procedure for nucleoside analogs of 6FP and prepared 1-(beta-d-ribofuranosyl)-2-(N,N diethylaminomethyleneamino)-6-formylpteridin-4-one (RDEF) and 1-(beta-d ribofuranosyl)-2-(piperidine-1-ylmethyleneamino)-6-formylpteridin-4-one (RPIF) in which the 1-position of 6FP is glycosylated. At pH 7.4, NADH was spontaneously oxidized to NAD(+) in the presence of RDEF in the dark. Using electron paramagnetic resonance analysis coupled with the spin trapping technique, we show that O(2) was converted to H(2)O(2)via superoxide anion radical ( O(2)(-)) during this reaction. The modification of the 1-position of 6FP did not cancel ROS generation activities, which were demonstrated in 3-position-modified 6FPs. Since the 6FP derivatives developed in the present study have a ribose moiety, these compounds can be subjected to further derivatization, such as incorporation into oligonucleotides, oligosaccharides, proteins, or any other compounds that recognize and interact with specific biomolecules, and therefore would be useful in pharmaceutical investigations that need generation of appropriate and controllable amounts of ROS in vivo. PMID- 17912386 TI - Stereoselective synthesis of 2-dienyl-substituted piperidines using an eta4 dienetricarbonyliron complex as the stereocontrolling element in a double reductive amination cascade. AB - In the presence of NaBH(OAc)(3), a 1,5-keto-aldehyde, contained within a side chain of an eta(4)-dienetricarbonyliron complex, undergoes a double reductive amination sequence with a series of primary amines, to provide the corresponding piperidine products in good to excellent yield. The dienetricarbonyliron complex functions as a powerful chiral auxiliary in this cascade process, exerting complete control over the stereoselectivity of the reaction, with the formation of a single diastereoisomeric product. The sense of stereoinduction has been confirmed by X-ray crystallography. Removal of the tricarbonyliron moiety can be effected with CuCl(2) to afford the corresponding 2-dienyl-substituted piperidine in excellent yield. Attempted extension of this cyclisation strategy to the corresponding azepane ring system using a 1,6-keto-aldehyde as the cyclisation precursor was unsuccessful; in this case, the reaction stopped after a single reductive amination on the aldehyde to provide an acyclic keto-amine product. PMID- 17912387 TI - D-glucosamine trimethylene dithioacetal derivatives: formation of six- and seven membered ring amino carbasugars. Synthesis of (-)-calystegine B3. AB - By virtue of carefully chosen protecting groups, d-glucosamine trimethylene dithioacetal derivatives were successfully oxidized to the corresponding 6 aldehydes. This methodology reverses the donor and acceptor position on a normal open chain sugar and changes the relative position of the N-substituent. From the 6-aldehydes, heptose epoxide derivatives were prepared by a Corey-Chaykovsky reaction, and cyclized by the Corey-Seebach method. Depending on the designed protecting groups, the orthogonally protected six- and seven-membered ring amino carbasugars can be produced selectively and efficiently. (-)-Calystegine B(3) was synthesized from one of those products with high yield. This is the first anionic cyclization pathway to calystegine type structures. PMID- 17912388 TI - Efficient asymmetric hydrogenation of olefins with hydrazine-derived diphosphoramidites. AB - Enantiopure, BINOL-derived diphosphoramidites built upon an achiral hydrazine spacer are efficient ligands for the hydrogenation of 2-(acetylamino)-3-(aryl) propenoic methyl esters. The activity and enantioselectivity of the hydrazine derivatives were shown to be markedly influenced by the nature of the two NR substituents, symmetrical but bulky R groups leading to the best results. A diphosphosphoramidite obtained from (t)BuHNNH(t)Bu resulted in ee's as high as 95%. The present results contradict previous reports on "short" diphosphoramidites. PMID- 17912390 TI - Intramolecular energy transfer in a tetra-coumarin perylene system: influence of solvent and bridging unit on electronic properties. AB - The synthesis and characterisation of a novel coumarin donor-perylene bisimide acceptor light-harvesting system is reported, in which an energy-transfer efficiency of >99% is achieved. Comparison of the excited-state properties of the donor-acceptor system with model compounds revealed that although the photophysical properties of the perylene bisimide acceptor unit are affected considerably by the nature of the substituent at the imide positions and the solvent employed, through-bond interaction between the donor and acceptor units is negligible. Energy transfer in the present system can be described as occurring via a through-space energy-transfer mechanism. Careful consideration of the redox properties of the donor relative to the acceptor units allows for avoidance of potentially deleterious excited-state electron-transfer processes. PMID- 17912389 TI - Enhanced aqueous solubility of long wavelength voltage-sensitive dyes by covalent attachment of polyethylene glycol. AB - Long wavelength voltage-sensitive dyes (VSDs) called Pittsburgh (PGH) dyes were recently synthesized by coupling various heterocyclic groups to a styryl thiophene intermediate forming extended, partially rigid chromophores. Unlike most styryl VSDs, dyes with a sulfonic acid anchor directly attached to the chromophore showed no solvatochromic absorption shifts. The limited water solubility of many long wavelength VSDs requires the use of surfactants to transport the dye through physiological saline solutions and effectively label biological membranes. Here, we tested the chemical substitution of the sulfonic acid moiety with polyethyleneglycol (PEG) chains, ranging from MW 750 to 5000, to overcome the poor solubility of VSDs while retaining their properties as VSDs. The chemical synthesis of PGH dyes and their PEG derivatives are described. The PEG derivatives were soluble in aqueous solutions (>1 mM) and still reported membrane potential changes. In frog and mouse hearts, the voltage sensitivity (DeltaF/F per action potential) and spectral properties of PEG dyes were the same as the sulfonated analogues. Thus, the solubility of VSDs can be considerably improved with small polyethyleneglycol chains and can provide an effective approach to improve staining of excitable tissues and optical recordings of membrane potential. PMID- 17912392 TI - Assembly of nanosize metallic particles and molecular wires on electrode surfaces. AB - This article highlights recent developments in the assembly of nanosize materials on electrode surfaces. A brief historical background of the field is given, followed by a selection of topics of particular current interest. We focus especially on the assembly of nanosize metallic particles and molecular wires on gold and silicon electrode surfaces. The fabrication, properties, and characteristics of functional nanostructured biointerfaces on electrode surfaces are also described. PMID- 17912391 TI - Glutathione traps formaldehyde by formation of a bicyclo[4.4.1]undecane adduct. AB - Glutathione forms complex reaction products with formaldehyde, which can be further modified through enzymatic modification. We studied the non-enzymatic reaction between glutathione and formaldehyde and identified a bicyclic complex containing two equivalents of formaldehyde and one glutathione molecule by protein X-ray crystallography (PDB accession number 2PFG). We have also used (1)H, (13)C and 2D NMR spectroscopy to confirm the structure of this unusual adduct. The key feature of this adduct is the involvement of the gamma-glutamyl alpha-amine and the Cys thiol in the formation of the bicyclic ring structure. These findings suggest that the structure of GSH allows for bi-dentate masking of the reactivity of formaldehyde. As this species predominates at near physiological pH values, we suggest this adduct may have biological significance. PMID- 17912393 TI - Spoilt for choice: assessing phase behavior models for the evolution of homochirality. AB - The observation of single chirality in biological systems has intrigued scientists for more than one hundred years. Here we discuss several recent experimental studies showing amplification of enantiomeric excess based on amino acid phase behavior. Comparing observations of solution-solid and gas-solid phase transitions highlights the underlying fundamental physical chemistry principles that rationalize the observed enantioenrichment in both cases. PMID- 17912394 TI - A discrete conglomerate of a distorted Mo(v)-porphyrin with a directly coordinated Keggin-type polyoxometalate. AB - The reaction of a saddle-distorted Mo(v)-dodecaphenylporphyrin complex and a Keggin-type polyoxometalate gives a discrete and nanosized molecule, [{Mo(DPP)(O)}(2)(H(2)SiW(12)O(40))], which involves direct coordination between the Mo(v) centers and terminal oxo groups of the polyoxometalate and exhibits excellent stability in solution to show reversible multi-redox processes. PMID- 17912395 TI - A self-assembled receptor for the recognition of phosphate and acetate anions in neutral aqueous solution. AB - The reaction of [(benzene)RuCl(2)](2) with a piperazine-bridged bis(dihydroxypyridine) ligand and LiOH leads to the formation of an expanded helicate, which is able to bind phosphate and acetate in aqueous solution at neutral pH. PMID- 17912396 TI - Examination of intermolecular electronic interactions in the crystal structure of C60(CF3)12 by experimental electron density determination. AB - From a high resolution X-ray data set measured at 20 K the experimental electron density of the fullerene C(60)(CF(3))(12) was derived and topologically analyzed to yield, in addition to bond topological and atomic properties, information about the density distribution in the region where hexagons of adjacent molecules approach closely at only 3.3 A. PMID- 17912397 TI - Uranyl stabilized Schiff base complex. AB - Uranyl Schiff base complex [(UO(2))(2)(Salpro)(OH)(Solvent)(2)] (1) in the presence of excess of ethylenediamine (EDA) does not undergo nucleophilic addition (hydrolysis) and substitution (transamination) reactions due to an extended chelation [2N, 3O + OH] by the flexible backbone. PMID- 17912398 TI - Zwitterionic pi-radical involving EDT-TTF-imidazole and F4TCNQ: redox properties and self-assembled structure by hydrogen-bonds and multiple S...S interactions. AB - The reaction between an imidazole-functionalized EDT-TTF and F(4)TCNQ produced a zwitterionic pi-radical, which formed a self-assembled structure by the cooperation of hydrogen-bonds and multiple S...S interactions and exhibited three step oxidation processes and a high electrical conductivity as a single-component organic molecule. PMID- 17912399 TI - Enantioselective catalysis with tropos ligands in chiral ionic liquids. AB - Enantioselective homogeneous rhodium-catalysed hydrogenation using tropoisomeric biphenylphosphine ligands was accomplished in readily available chiral ionic liquids and the catalytic system could be reused after extraction with scCO(2). PMID- 17912400 TI - Sol-gel encapsulation of cells is not limited to silica: long-term viability of bacteria in alumina matrices. AB - A colloidal route to aqueous alumina gels is described, allowing the long-term viability of encapsulated bacteria. PMID- 17912401 TI - Area-selective growth of amorphous carbon nanofibers via catalytic decomposition of polyimide thin film. AB - A dense layer of amorphous carbon nanofibers was fabricated by pyrolyzing a thin film of polyimide using a monolayer of gold nanoparticles as a catalyst. PMID- 17912403 TI - Competitive O-H and C-H oxidative addition of CH3OH to rhodium(II) porphyrins. AB - Rhodium(II) porphyrins react with CH(3)OH in benzene by alternate mechanisms that give H-CH(2)OH and H-OCH(3) bond activation in different methanol concentration regimes which is a rare example of transition metal reactivity with methanol. PMID- 17912402 TI - Inorganic polymer photoresist for direct ceramic patterning by photolithography. AB - A novel negative, inorganic polymer photoresist was demonstrated to be suitable for simple and direct fabrication of tribological SiCN-based ceramic microstructures via UV photolithography and subsequent pyrolysis at 800 degrees C. PMID- 17912405 TI - Diastereoselective Simmons-Smith cyclopropanations of allylic amines and carbamates. AB - Cyclopropanation of 3-(N,N-dibenzylamino)cyclohexene with either Zn(CH(2)I)(2)(Wittig-Furukawa reagent) or CF(3)CO(2)ZnCH(2)I (Shi's reagent) gives the corresponding syn-cyclopropane as a single diastereoisomer, whilst cyclopropanation of 3-(N-tert-butoxycarbonylamino)cyclohexene with CF(3)CO(2)ZnCH(2)I gives the corresponding anti-cyclopropane exclusively; facile N-deprotection gives access to either diastereoisomer of the trifluoroacetic acid salt of 2-aminobicyclo[4.1.0]heptane. PMID- 17912404 TI - Evaluation of a carbohydrate-pi interaction in a peptide model system. AB - A carbohydrate-pi interaction contributes -0.8 kcal mol(-1) to the stabilization of a beta-hairpin peptide. PMID- 17912406 TI - Superacid promoted reactions of N-acyliminium salts and evidence for the involvement of superelectrophiles. AB - Experimental and theoretical studies suggest the involvement of dicationic, superelectrophilic N-acyliminium ions in conversions catalyzed by superacids. PMID- 17912407 TI - Beta-chlorovinylsilanes as masked alkynes in oligoyne assembly: synthesis of the first aryl-end-capped dodecayne. AB - An aryl-end-capped dodecayne has been prepared using a four-fold fluoride mediated dechlorosilylation of a masked dodecayne precursor containing four beta chlorovinylsilane residues that serve as masked alkynes; the unstable dodecayne product has been characterised by UV-vis absorption spectroscopy and 'matrix free' MALDI-TOF mass spectrometry. PMID- 17912408 TI - Highly efficient asymmetric reduction of arylpropionic aldehydes by horse liver alcohol dehydrogenase through dynamic kinetic resolution. AB - The enantioselective synthesis of (2S)-2-phenylpropanol and (2S)-2-(4-iso butylphenyl)propanol ((S)-Ibuprofenol) has been achieved by means of Horse Liver Alcohol Dehydrogenase (HLADH) in buffered aqueous solution or buffered organic solvent mixtures; under the reaction conditions, a dynamic kinetic resolution (DKR) process was realized with good reaction yields and enantiomeric ratios. PMID- 17912409 TI - Microwave-assisted preparations of dihydropyrroles from alkenone O-phenyl oximes. AB - Microwave irradiation of alkenone O-phenyl oximes produces iminyl radicals that ring close to yield dihydropyrrole derivatives; pyrroles and pyridines can be obtained from related precursors. PMID- 17912410 TI - An esterase-activated magnetic resonance contrast agent. AB - A Gd(III) complex bearing pendant acetoxymethyl esters is activated on exposure to porcine liver esterase; the 84% increase in relaxivity is a result of suppression of HCO(3)(-)/CO(3)(2-) binding by the resulting negative charge. PMID- 17912411 TI - Immobilization of nanofibrous metal oxides on microfibers: a macrostructured catalyst system functionalized with nanoscale fibrous metal oxides. AB - Nanofibrous LaMnO(3) can be immobilized on macrostructured materials using carbon nanofibers as templates; their application as macro-nanostructured catalysts are also presented. PMID- 17912412 TI - Control of ionic conductivity of ionic liquid/photoresponsive poly(amide acid) gels by photoirradiation. AB - 1,3-Dibutylimidazolium bromide was soaked into a polymer network prepared by condensation of 4-4'-diaminoazobenzene, pyromellitic dianhydride and 1,3,5-tri(4 aminophenyl)benzene to form photoresponsive ion conductive gels. PMID- 17912414 TI - Consecutive palladium-catalyzed Hiyama-Heck reactions in aqueous media under ligand-free conditions. AB - Symmetric and asymmetric (E)-1,2-diarylethenes are synthesized from aryl bromides by consecutive one-pot Hiyama-Heck reactions carried out in water and under air; the only additives required are sodium hydroxide, palladium acetate and poly(ethylene glycol), and the products are isolable in many cases by simple filtration of the water solution. PMID- 17912413 TI - Characterisation of 3-methylorcinaldehyde synthase (MOS) in Acremonium strictum: first observation of a reductive release mechanism during polyketide biosynthesis. AB - Isolation and sequencing of a PKS gene isolated from xenovulene-producing cultures of Acremonium strictum indicated the presence of NT-, KS-, AT-, PT-, C MeT- and R-domains; heterologous expression in Aspergillus oryzae resulted in the production of 3-methylorcinaldehyde, demonstrating the role of the terminal reductase domain in product release. PMID- 17912415 TI - Structural and dynamical properties of guest molecules confined in mesoporous silica materials revealed by NMR. AB - In the last fifteen years several novel porous silica materials, which are periodically structured on the mesoscopic length scale, have been synthesized. They are of broad interest for fundamental studies of surface-substrate interactions, for studies of the dynamics of guest molecules in confinement and for studies of the effect of confinement on the structural and thermophysical properties of fluids. Examples of such confinement effects include the change of the freezing and melting points or glass transitions of the confined liquids. These effects are studied by combinations of several NMR techniques, such as (15)N- and (2)H-solid-state NMR line shape analysis, MAS NMR and NMR diffusometry with physico-chemical characterization techniques such as nitrogen adsorption and small angle diffraction of neutrons or X-rays. This combination does not require crystalline samples or special clean and well defined surfaces such as conventional surface science techniques, but can work with typical ill-defined real world systems. The review discusses, after a short introduction, the salient features of these materials and the applied NMR experiments to give the reader a basic knowledge of the systems and the experiments. The rest of the review then focuses on the structural and dynamical properties of guest molecules confined in the mesoporous silica. It is shown that the confinement into the pores leads to fascinating new features of the guests, which are often not known for their bulk phases. These features depend strongly on the interplay of the their interactions with the silica surface and their mutual interactions. PMID- 17912416 TI - Reactivity of Ti(IV) species hosted in TS-1 towards H2O2-H2O solutions investigated by ab initio cluster and periodic approaches combined with experimental XANES and EXAFS data: a review and new highlights. AB - This work is intended to underline how the most-advanced experimental and theoretical physical chemistry tools must be used synergistically to understand the reactivity of Ti-silicalite-1 (TS-1) in an important number of low temperature oxidation reactions with aqueous H(2)O(2) as the oxidant. Literature results are carefully reviewed and accompanied with new, unpublished highlights of both experimental and computational origin. The first part of this work is devoted to a discussion of the defective nature of the material and to the synergic role played by Si vacancies and Ti insertion in the framework. A summary of the experimental Ti-K-edge EXAFS and XANES literature concerning the activated material in vacuo conditions is then presented and compared to the corresponding Ti geometries obtained by ab initio calculations. From such a comparison, the excellent agreement between experiment and theory is evident. A very good agreement is also obtained for the interaction with water and ammonia. For both H(2)O and NH(3), the agreement is due to the possibility to perform experiments in which the probe molecule is dosed from the gas phase, thus allowing to reach the 1 : 1 (or 1 : 2) ratio between the adsorbing Ti sites and the adsorbed molecule. Then, interaction with hydrogen peroxide is discussed, underlining the problems faced in reaching a common view between experimental and theoretical results, owing to the difficulties both in performing experiments where anhydrous H(2)O(2) is dosed on TS-1, and in taking into account the role played by the aqueous medium in the reactivity of Ti(iv) centres toward H(2)O(2) using ab initio calculations. Only once such difficulties have been overcome, by increasing the complexities of both experiments and ab initio models, is a joint view finally obtained. Where needed, comparison with other experimental results (X-ray and neutron diffraction, NMR, IR, Raman, UV-vis and resonant Raman) is made. PMID- 17912417 TI - Solid-state NMR of endohedral hydrogen-fullerene complexes. AB - We present an overview of solid-state NMR studies of endohedral H(2)-fullerene complexes, including (1)H and (13)C NMR spectra, (1)H and (13)C spin relaxation studies, and the results of (1)H dipole-dipole recoupling experiments. The available data involves three different endohedral H(2)-fullerene complexes, studied over a wide range of temperatures and applied magnetic fields. The symmetry of the cage influences strongly the motionally-averaged nuclear spin interactions of the endohedral H(2) species, as well as its spin relaxation behaviour. In addition, the non-bonding interactions between fullerene cages are influenced by the presence of endohedral hydrogen molecules. The review also presents several pieces of experimental data which are not yet understood, one example being the structured (1)H NMR lineshapes of endohedral H(2) molecules trapped in highly symmetric cages at cryogenic temperatures. This review demonstrates the richness of NMR phenomena displayed by H(2)-fullerene complexes, especially in the cryogenic regime. PMID- 17912418 TI - The pure rotational spectrum of the actinide-containing compound thorium monoxide. AB - The J = 1-0 pure rotational transition, together with hyperfine structure where appropriate, has been recorded for all three naturally occurring isotopomers of the actinide-containing compound thorium monoxide ((232)Th(16)O, (232)Th(17)O and (232)Th(18)O). PMID- 17912419 TI - Photoswitching electron transport properties of an azobenzene containing thiol SAM. AB - The influence of conformational and electrical properties of azobenzene molecules on the electron transfer barrier properties of their SAMs was studied by SECM and ellipsometry. PMID- 17912420 TI - Catalytic oxidation of methanol on Pd metal and oxide clusters at near-ambient temperatures. AB - Supported Pd clusters catalyze methanol oxidation to methyl formate with high turnover rates and >90% selectivity at near ambient temperatures (313 K). Metal clusters are much more reactive than PdO clusters and rates are inhibited by the reactant O(2). These data suggest that ensembles of Pd metal atoms on surfaces nearly saturated with chemisorbed oxygen are required for kinetically-relevant C H bond activation in chemisorbed methoxide intermediates. Pd metal surfaces become more reactive with increasing metal particle size. The higher coordination of surface atoms on larger clusters leads to more weakly-bound chemisorbed species and to a larger number of Pd metal ensembles available during steady state catalysis. Chemisorbed oxygen removes H-atoms formed in C-H bond activation steps and inhibits methoxide decomposition and CO(2) formation, two functions essential for the high turnover rates and methyl formate selectivities reported here. PMID- 17912421 TI - A Monte Carlo-quantum mechanics study of the lowest n-pi* and pi-pi* states of uracil in water. AB - The solvatochromic shifts of the n-pi(*) and pi-pi(*) states of uracil in water are analyzed using a combined and sequential Monte Carlo/quantum mechanics (MC/QM) approach. The role of the solute polarization and electronic delocalization into the solvent region are investigated. Electronic polarization of the solute is obtained using the HF/6-31G(d), the polarizable continuum model (PCM) and an iterative procedure using MP2/aug-cc-pVDZ in the MC/QM. The in-water dipole moment of uracil is obtained, respectively, as 5.12 D, 6.12 D and 7.01 +/- 0.05 D. This latter result, corresponding to an increase of 60% with respect to the gas phase value, is used in the classical potential of the MC simulation to obtain statistically uncorrelated configurations for subsequent QM calculations of the ultraviolet-visible absorption spectrum of uracil in water. QM calculations are performed at the time-dependent density-functional theory (TD DFT) combined with the B3LYP and B3PW91 functionals, multiconfigurational (CASSCF) and the semi-empirical all-valence electron INDO/CIS methods. Using 60 solute-solvent configurations with the explicit inclusion of 200 water molecules the solvatochromic shift is obtained as a blue shift of 0.50 eV for the n-pi(*) state and a red shift of 0.19 eV for the pi-pi(*) state, in good agreement with experimentally-inferred values. These results are compared with TD-DFT results in conjunction with PCM approaches and the importance of solute polarization and wave function delocalization over the solvent region is discussed. Our results suggest that the elusive n-pi(*) state of uracil in water lies around 255 nm hidden by the intense and broad pi-pi(*) transition with a maximum at 260 nm, inverting the relative locations of these states compared to the gas phase. This is further supported by considering the in-water dipole moment changes upon excitation, as obtained from CASSCF calculations. PMID- 17912423 TI - Enhancement factor averaging and the photostability of probes in SERS vibrational pumping. AB - The technique of temperature dependent vibrational pumping in surface enhanced Raman scattering (SERS) has been recently demonstrated as a promising new tool to estimate SERS cross-sections. In this paper we expand on the previous developments and study several details around the implementation and physics of the vibrational pumping technique in SERS. Here we concentrate on two specific aspects related to: (i) the different averaging properties (over the distribution of enhancements) of the Stokes and anti-Stokes signals in the pumping regime; and (ii) the role of the finite photostability of the probes. The fact that the anti Stokes signal is averaged differently from the Stokes counterpart leads to some unique phenomena in Raman spectroscopy that can only be observed under the conditions of vibrational pumping in SERS. PMID- 17912422 TI - The Gaussian Generalized Born model: application to small molecules. AB - This work presents a Generalized Born model for the computation of the electrostatic component of solvation energies which is based on volume integration. An analytic masking function is introduced to remove Coulombic singularities. This approach leads to analytic formulae for the computation of Born radii, which are differentiable to arbitrary order, and computationally straightforward to implement. PMID- 17912424 TI - Oxide thin films based on ordered arrays of 1D nanostructure. A possible approach toward bridging material gap in catalysis. AB - TiO(2) thin films based on ordered arrays of 1D nanostructures (nanorods, nanotubes) are proposed as suitable model materials in studies for bridging material and complexity gap in catalysis. The samples were prepared by anodic oxidation of Ti foils. By changing the preparation conditions (pH, procedure of application of the potential), different types of 1D nanostructure and different characteristics of the ordered array of these 1D nanostructures could be obtained. This allows studying the effect of nanodimension and 3D nanoarchitecture on the characteristics and reactivity of these catalysts. It is also shown that TiO(2) thin films characterized by a well-ordered array of titania nanorod behave as photonic materials, thus showing unique properties of light harvesting efficiency. PMID- 17912426 TI - Host mediated anti-tumor effect of oncolytic Newcastle disease virus after locoregional application. AB - Several strains of the Newcastle disease virus (NDV) have raised considerable interest in recent years for clinical application because of their oncolytic properties. In this study we characterized virological, immunological and anti tumor properties of some NDV strains. The oncolytic strain MTH-68/H was the most potent interferon-alpha inducer and, after UV light inactivation, it was the only tested NDV strain which induced in human PBMC anti-tumor activity in vitro. Upon systemic application to mice bearing a virus susceptible intradermal tumor, no significant anti-tumor effects were observed with the two oncolytic strains Italian and MTH-68/H while the treatment had significant side effects as seen by loss of body weight. In contrast, when using a locoregional application model for treatment of liver metastases of luciferase transfected CT26 colon carcinoma cells, MTH-68/H showed a significant delay in tumor growth, as well as prolonged survival but no effects on body weight. Surprisingly, this CT26 murine tumor cell transfectant was resistant in vitro to virus infection and oncolysis. These results suggest: i) that locoregional application of oncolytic NDV is more effective than systemic i.v. application; and ii) that oncolytic NDV can mediate effects even against a virus-resistant tumor line. The involvement of host anti tumor immune responses as an important mechanism in therapies based on oncolytic NDV will be discussed. PMID- 17912425 TI - mda-7/IL-24, novel anticancer cytokine: focus on bystander antitumor, radiosensitization and antiangiogenic properties and overview of the phase I clinical experience (Review). AB - Subtraction hybridization applied to a 'differentiation therapy' model of cancer employing human melanoma cells resulted in the cloning of melanoma differentiation associated gene-7/interleukin-24 (mda-7/IL-24). Initial studies confirm an inverse correlation between mda-7 expression and melanoma development and progression. Forced expression of mda-7 by means of a plasmid or via a replication incompetent adenovirus (Ad.mda-7) promotes growth suppression and induces apoptosis in a broad array of human cancers. In contrast, mda-7 does not induce growth suppressive or toxic effects in normal cells. Based on structure (containing an IL-10 signature motif), secretion by cells (including subsets of T cells) and location on chromosome 1q (in an area containing IL-10-family genes), mda-7 has now been renamed mda-7/IL-24. Studies by several laboratories have uncovered many of mda-7/IL-24's unique properties, including cancer-specific apoptosis-induction, cell cycle regulation, an ability to inhibit angiogenesis, potent 'bystander antitumor activity' and a capacity to enhance the sensitivity of tumor cells to radiation, chemotherapy and monoclonal antibody therapy. Moreover, based on its profound cancer tropism, substantiated by in vivo human xenograft studies in nude mice, mda-7/IL-24 (administered as Ad.mda-7) was evaluated in a phase I clinical trial in patients with melanomas and solid cancers. These studies document that mda-7/IL-24 is well tolerated and demonstrates evidence of significant clinical activity. In these contexts, mda 7/IL-24 represents a unique cytokine gene with potential for therapy of human cancers. The present review focuses on three unique properties of mda-7/IL-24, namely its potent 'bystander antitumor activity', ability to sensitize tumor cells to radiation, and its antiangiogenesis properties. Additionally, an overview of the phase I clinical trial is provided. These studies affirm that mda 7/IL-24 has promise for the management of diverse cancers. PMID- 17912427 TI - Overexpression of Septin1: possible contribution to the development of oral cancer. AB - Our previous study using proteomic profiling demonstrated significant up regulation of Septin1, a conserved family of GTPase proteins, in oral squamous cell carcinoma (OSCC)-derived cell lines. In the current study, to determine the potential involvement of Septin1 in oral carcinogenesis, we evaluated the state of septin1 protein/mRNA expression in OSCC-derived cell lines, oral premalignant lesions (OPLs), and primary OSCCs. A significant (P<0.05) increase in Septin1 expression was evident in all OSCC-derived cell lines examined compared to human normal oral keratinocytes (HNOKs) and OPLs. In immunohistochemistry, while the vast majority of the OSCCs (89%) were positive for Septin1, no immunoreaction was observed in corresponding normal tissues and OPLs. In addition, real-time quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) data were consistent with the protein expression status. These results suggest that Septin1 expression could contribute to cancer progression, proliferation, or both, and that Septin1 may be a potential diagnostic marker of highly active cancer and a therapeutic target for OSCCs. PMID- 17912428 TI - Expression profiling of fecal colonocytes for RNA-based screening of colorectal cancer. AB - The early detection of colorectal cancer originating from any part of the colorectum is desirable because this cancer can be cured surgically if diagnosed early. We searched for marker genes for a fecal RNA-based colorectal cancer screening method by comparison of genome-wide expression profiles among cancerous and non-cancerous tissues, and healthy volunteer- and cancer patient-derived colonocytes from the feces, and the peripheral blood. Of 14,564 genes, only 3 (PAP, REG1A, and DPEP1) were selectable as final candidates which were expressed frequently at any stage of this cancer and were suppressed in non-cancerous tissues and also in the peripheral blood and colonocytes of healthy volunteers. Next, we directly compared fecal RNA-expression profiles between colorectal cancer patients and healthy volunteers, and found that most of the genes (92%) expressed in the colonocytes of the cancer patients were not expressed in those of the healthy volunteers. Six genes (SEPP1, RPL27A, ATP1B1, EEF1A1, SFN, and RPS11) selected randomly from 85 cancer patient-derived colonocyte-specific genes were evaluated. In total, reverse transcription-polymerase chain reaction or focused microarray of all those 9 genes detected 18 (78%) of 23 curable colorectal cancers (Dukes stages A-C), 9 or 10 (64% or 71%) of 14 early cancers with no lymph node metastasis (Dukes stage A or B) and 4 (80%) of 5 right-sided cancers. Our extensive gene list provides other markers for fecal RNA-based colorectal cancer screening. PMID- 17912430 TI - Mutations and differential expression of the ras family genes in human nasal polyposis. AB - Although it is well established that ras genes contribute to tumourigenesis either through the accumulation of mutations or by aberrant expression in a wide range of human cancers, little is known regarding their involvement in human nasal polyps (NPs). In the present study, the occurrence of mutations in codons 11 and 12 of the ras family genes was examined by PCR/RFLP and direct sequencing in 23 human NPs and their adjacent turbinates, as well as in turbinates from 13 control subjects. Moreover, the expression pattern of ras mRNA levels was assessed in NP specimens and compared to adjacent and control tissues. K-ras codon 11 and 12 mutations were detected in 17 and 35% of NPs, respectively, and were found in the adjacent inferior turbinate (AIT) (22 and 16%, respectively) and adjacent middle turbinates (AMT) (16 and 26%, respectively). K- and H-ras expression levels were elevated, whereas N-ras mRNA levels were lower in NPs and adjacent turbinates as compared to the control tissues. K-ras mRNA levels were up regulated in advanced-stage polyps (P=0.037), while N-ras levels were found elevated in small polyps (P=0.046). Statistically significant negative correlations between K- and N-ras expression profiles arose in NPs and AITs (P=0.009 and 0.003, respectively). This, to our knowledge, is the first report on ras mutations and expression analysis in NPs. Our findings suggest a potential key role for activated members of ras family genes in terms of their contribution to the development of NPs as well as to the hypertrophy of adjacent turbinates. PMID- 17912431 TI - Mutation of the p16/CDKN2 gene and loss of heterozygosity in malignant mucosal melanoma and adenoid cystic carcinoma of the head and neck. AB - The purpose of this study was to investigate the molecular biological characteristics of malignant mucosal melanoma (MMM) and adenoid cystic carcinoma (ACC) of the head and neck. We analyzed the common genetic abnormalities that may help to identify the loci in the genes involved in the development of MMM and ACC of the head and neck by PCR-LOH on chromosomes 1p, 6q, 9p, 10q, 11q, 12q, 17p, and 19q. LOH was observed in 6 of 12 cases of MMM and in 12 of 15 cases of ACC informative for at least one of the loci analyzed. One distinct deleted region was identified at chromosome 9p21. In addition, to identify a possible involvement of p16/CDKN2 PCR-SSCP and auto-DNA sequence analysis were also performed to detect any mutation of the p16/CDKN2. Particularly, 2 missense mutations were detected in codon 225 and 226, both in MMM and ACC. There were mutational hot spots in the p16/CDKN2 gene. These results suggested that mutation of the p16/CDKN2 gene was a common factor in the development of human MMMs and ACCs, while this gene may be correlated with development and/or progression of a subtype and play a role in the oncogenesis of these cancers. PMID- 17912429 TI - RNAi-mediated abrogation of cathepsin B and MMP-9 gene expression in a malignant meningioma cell line leads to decreased tumor growth, invasion and angiogenesis. AB - Malignant meningiomas are highly aggressive and frequently recur after surgical resection of the tumor. Earlier studies have reported that the cysteine protease cathepsin B and the matrix metalloproteinase MMP-9 play important roles in tumor progression. In the present study, we made an attempt to evaluate the roles of these proteases in the malignant meningioma tumor microenvironment and determined the effectiveness of using single or bicistronic siRNA constructs for cathepsin B and MMP-9, in both in vitro and in vivo models. Transfection of a plasmid vector expressing double-stranded RNA for cathepsin B and MMP-9 significantly inhibited mRNA and protein levels of cathepsin B and MMP-9. The migration and invasion of meningioma cells were decreased after treatment with single or bicistronic siRNA constructs for cathepsin B and MMP-9 compared to controls and vector controls. Inhibition of angiogenesis was observed when the cells were transfected with single or bicistronic constructs for cathepsin B and MMP-9, when compared to controls or empty vector controls. Our study revealed that abrogation of cathepsin B and MMP-9 expression decreased the activation of major proteins involved in MAP kinase and PI3 kinase pathways indicating that targeting these proteases may hinder intracellular signaling and thus decrease cell survival and proliferation in malignant meningiomas. In addition to the in vitro evidence, we observed a significant regression of pre-established orthotopic tumors after treatment with RNAi plasmid vectors targeting cathepsin B and MMP-9. Furthermore, these observations demonstrate that the simultaneous RNAi-mediated targeting of cathepsin B and MMP-9 has potential application for the treatment of human meningiomas. PMID- 17912432 TI - Methylthioadenosine phosphorylase deficiency in Japanese osteosarcoma patients. AB - Methylthioadenosine phosphorylase (MTAP) is an important enzyme in the salvage pathway of adenosine and methionine synthesis. MTAP is ubiquitously present in all normal cells and tissues, but deficient in a variety of malignant tumors. The enzyme deficiency is caused by either MTAP gene deletion or promoter hypermethylation. We investigated MTAP expression, MTAP gene deletion and promoter abnormality in 40 primary tumor samples from Japanese osteosarcoma patients and determined the frequency of the enzyme deficiency. We also tested whether or not the enzyme deficiency can be exploited for tumor-specific chemotherapy using osteosarcoma cell lines. For MTAP expression, immunohistochemistry (IHC) and Western blotting were used. Real-time quantitative PCR assay was used for the analysis of MTAP gene deletion in fifteen osteosarcoma samples. MTAP promoter abnormality was analyzed by methylation-specific PCR. Then, the relationship between MTAP expression and sensitivity to the inhibitors of de novo AMP synthesis was confirmed in an MTAP-negative and -positive osteosarcoma cell line. The MTAP protein was negative in 11 of 40 samples (27.5%) by IHC and in 4 of 6 osteosarcoma cell lines (66.7%) by Western blot analysis. Among 40 samples, 15 were subjected to quantitative real-time PCR and promoter methylation analysis. Of 6 samples that were negative by IHC, the MTAP gene was deleted in 3 and the MTAP promoter was methylated in 2. These results indicated that MTAP deficiency was caused by MTAP gene deletion or promoter methylation in all MTAP-negative samples except one that was negative with IHC although no deletion or promoter methylation was detected. In in vitro experiments using transfectoma along with the MTAP-negative parental cell line, the MTAP-negative parental cell line was more chemosensitive to the inhibitors of de novo AMP synthesis than MTAP-positive transfectoma. MTAP deficiency frequently found in osteosarcoma can be exploited for selective chemotherapy in MTAP-negative osteosarcoma patients with the inhibitors of de novo purine synthesis. PMID- 17912433 TI - Prophylactic inguinal-femoral irradiation as an alternative to primary lymphadenectomy in treatment of vulvar carcinoma. AB - In a complete geographic series of 294 cases of primary vulvar carcinomas prophylactic inguinal-femoral irradiation was used as a standard postoperative therapy. Inguinal lymph node dissection was performed in only 27 cases (9%) and was not part of the standard surgery. The histology was squamous cell carcinoma in 269 cases (92%). The primary surgery was total vulvectomy, partial vulvectomy, or local resection of the tumor. The main type of radiotherapy was adjuvant inguinal irradiation. Two separate, symmetrical and rectangular inguinal fields were irradiated with combined photon and electron beams. In the complete series 127 recurrences (43%) were recorded. Local (24%) and regional recurrences (19%) were most frequent. Type of surgery was not associated with the risk of tumor recurrence. The 5-year overall survival rate was 53% and the relapse-free survival (RFS) rate was 55%. Tumor grade was significantly (P=0.007) associated with the RFS. The inguinal RFS rate was 75% both for patients treated with adjuvant inguinal irradiation without lymphadenectomy and patients treated with primary lymph adenectomy +/- inguinal irradiation. Postoperative complications were recorded in 22%. Postoperative complications occurred most frequently in the subgroup undergoing inguinal lymphadenectomy. Chronic lymph edemas were the most serious late tissue reactions. PMID- 17912434 TI - Combined effect of 2-5A-linked antisense against telomerase RNA and conventional therapies on human malignant glioma cells in vitro and in vivo. AB - We recently showed that therapy with 2'-5'-oligoadenylate (2-5A)-linked antisense against human telomerase RNA component (2-5A-anti-hTR) is a novel telomerase targeting strategy against malignant gliomas. In this study, we investigated conventional chemotherapeutic agents and gamma-irradiation (IR) to determine whether they could augment the efficacy of 2-5A-anti-hTR against these tumors in vitro and in vivo. Treatment with 2-5A-anti-hTR inhibited the viability of U373 MG and U87-MG malignant glioma cells in a dose-dependent manner; the antitumor effect resulted from induction of apoptosis. Also, telomerase-positive astrocytes with oncogenic Ras were more sensitive to 2-5A-anti-hTR than were those without oncogenic Ras. In addition, we sought to determine the combined effect of 2-5A anti-hTR with N, N'-bis (2-chloroethyl)-N-nitrosourea (BCNU), cisplatin (CDDP), paclitaxel (PTX), temozolomide (TMZ), or IR. When we administered the combination treatments on the same day, PTX and IR showed a greater combined effect with 2-5A anti-hTR on both tumor cell lines than did BCNU, CDDP and TMZ. However, all of the combination regimens were synergistic when we first treated tumor cells with 2-5A-anti-hTR for 24 h and then exposed them to the conventional treatments. Apoptosis-inducing agents (CDDP and PTX) but not autophagy-inducing therapies (TMZ and IR) enhanced the incidence of apoptosis caused by 2-5A-anti-hTR. Lastly, we observed a combinatorial effect of 2-5A-anti-hTR and TMZ in vivo in subcutaneous U87-MG tumors in nude mice. Interestingly, treatment with TMZ increased the incidence of apoptosis in subcutaneous tumor cells treated with 2 5A-anti-hTR. These results suggest that 2-5A-anti-hTR is preferable in combination with established cancer therapies. PMID- 17912435 TI - Clinical significance of growth differentiation factor 11 in colorectal cancer. AB - Growth differentiation factor 11 (GDF11), a member of the transforming growth factor-beta superfamily and bone morphogenetic protein (BMP) subfamily, plays a role in regulation of development and differentiation. Although some members of BMP subfamily have been reported to correlate with cancer, the significance of GDF11 has not been studied in a clinical oncology setting. The current study explored the clinicopathological significance of GDF11 expression in colorectal cancer. Quantitative real-time reverse transcription-PCR in colorectal cancer specimens obtained from 130 patients showed that GDF11 mRNA expression in cancer tissue was significantly higher than in normal tissue (p=0.001). Tumors were classified as high GDF11 expression (n=65) or low GDF11 expression (n=65). Patients whose tumors had high GDF11 expression showed a high frequency of lymph node metastasis (p=0.049) and had more cancer-related deaths (p=0.040). Furthermore, the patients with high GDF11 expression had significantly poorer overall survival than those with low expression (p=0.0334). Although multivariate analysis showed that GDF11 was not an independent prognostic factor, these findings suggest that GDF11 may be a novel diagnostic and prognostic biomarker in patients with colorectal cancer. PMID- 17912436 TI - Hypoxia-inducible expression of the mouse carbonic anhydrase IX demonstrated by new monoclonal antibodies. AB - CA IX is a transmembrane carbonic anhydrase isoenzyme predominantly expressed in human tumors in response to hypoxia and functionally implicated in adaptation of tumor cells to hypoxic stress via control of pH and cell adhesion. Intense investigations of the human CA IX as a hypoxic marker and a therapeutic target have been facilitated by specific monoclonal antibodies. However, no such reagents existed for the mouse CA IX ortholog. We generated five new anti-mouse CA IX monoclonal antibodies AM1-4, AM4-3, AM27-4, AM34-7 and AM35-1 produced using CA IX-deficient mice. The antibodies are suitable for various immunodetection methods including immunoblotting and immunohistochemistry. Using these reagents we show that the mouse CA IX is expressed in three out of nine tested mouse cell lines, namely in L929, MEF and TSA and is regulated by hypoxia and cell density similarly to human CA IX. We also demonstrate that the mouse CA IX exhibits hypoxia-related expression pattern in multicellular spheroids and in tumor xenografts. Our results indicate the use of the mouse model as suitable for further studies of CA IX role in tumor development and for its pre-clinical investigations. The new monoclonal antibodies represent potent tools for accomplishment of these future studies. PMID- 17912438 TI - CD44 crosslinking-mediated matrix metalloproteinase-9 relocation in breast tumor cells leads to enhanced metastasis. AB - CD44 plays a major role in multiple physiological processes, including cell-cell adhesion, cell-substrate interaction, lymphocyte homing, and tumor metastasis. It has been reported that highly expressed CD44 in certain types of tumors is associated with the hematogenic spread of tumor cells. The ability of CD44 to bind hyaluronan has been shown to correlate with tumor cell invasiveness, and it is likely that this ability may enhance tumor cell migration at several points during metastasis. However, the mechanism as to how CD44 stimulates metastasis remains unknown. The human breast tumor cell line, MDA-MB-435s, was used to investigate the effect of antibody-mediated CD44 crosslinking on the cellular level and localization of matrix metalloproteinase-9 (MMP-9). Confocal microscopy and immunocytochemical analyses were performed to demonstrate colocalization of CD44 and MMP-9 after CD44 crosslinking. Furthermore, the CD44-MMP-9 complex was purified by immunoprecipitation. G8 myoblast monolayers were employed to evaluate the invasiveness of human breast tumor cells after CD44 crosslinking in the presence or absence of protease inhibitors. CD44 crosslinking augmented the level of MMP-9 in the membrane of human breast tumor cells and clustering of CD44 serves as an MMP-9 docking molecule allowing MMP-9 to retain its concentrated proteolytic activity on the cell surface. Furthermore, crosslinking of CD44 enhances the ability of breast tumor cells to invade G8 myoblast monolayers and migrate through the basal membranes which was inhibited in the presence of anti MMP-9 antibody or the MMP inhibitors GM6001 or 1,10-phenanthroline. This study demonstrates for the first time that CD44 crosslinking leads to an enhanced level and relocation of MMP-9 in human breast tumor cells accompanied by increased tumor invasion and metastasis. PMID- 17912437 TI - Short hairpin RNA targeting survivin inhibits growth and angiogenesis of glioma U251 cells. AB - Survivin is a novel tumor-associated gene, its overexpression mostly associates with carcinogenesis and development. Nevertheless, the precise role of survivin in initiation and progression of gliomas is still not completely clear. We constructed here three short hairpin RNA (shRNA) targeting survivin plasmid vectors and introduced them into glioma U251 cells. The three shRNAs were efficiently and specifically able to knockdown the survivin expression in transiently transfected U251 cells. The stable transfectants expressing the shRNA having the strongest inhibitory effect against survivin exhibited decreased cell growth, increased spontaneous apoptosis, mitotic catastrophe and cell cycle arrest. Furthermore, in nude mice xenografts, the stable transfectants presented decreased de novo glioma formation and reduced development of angiogenesis. Results from this study indicate that survivin plays an important role in malignant proliferation, antiapoptosis and angiogenesis of gliomas, which may become an attractive target for gene therapy of gliomas, while RNA interference (RNAi) mediated by shRNA may become a new promising strategy for cancer gene therapy. PMID- 17912439 TI - Angiogenic characteristics of circulating and tumoural thrombospondin-1 in breast cancer. AB - In cancer models, thrombospondin-1 (TSP-1) has been shown to inhibit angiogenesis or promote metastasis by increasing adhesion of malignant cells to endothelium. To determine the role of TSP-1 in breast cancer and breast cancer angiogenesis, we have measured TSP-1 in plasma and tumour cytosols and compared levels to established clinicopathological prognostic parameters and intratumoural microvessel density. TSP-1 was measured, by radioimmunoassay, in plasma (pTSP-1) and tumour cytosols (cTSP-1) of women with early breast cancer (EBC) (n=71). pTSP 1 in EBC was compared to pTSP-1 levels in women with advanced breast cancer (ABC) (n=66), normal controls (n=77) and was correlated with prognostic features and microvessel density (MVD) (measured by CD31 immunostaining). cTSP-1 levels were compared to prognostic features and microvessel density. pTSP-1 in women with EBC (median 484, IQR 344-877 ng/ml) and ABC (median 588, IQR 430-952 ng/ml) were elevated when compared to normal controls (median 21, IQR 175-247) (p<0.001). Women with lymph node metastases (n=35) had higher levels of TSP-1 (median 799 ng/ml, IQR 455-943) than women who were node negative (median 343 ng/ml, IQR 267 514) (n=36) (p<0.05). Levels of pTSP-1 in EBC correlated with MVD (R=0.39, p<0.05). Levels of TSP-1 in tumour cytosols of women with EBC (median 1714, IQR 893-5283 ng/ml) correlated with microvessel density (R=0.46, p<0.01). Circulating levels of TSP-1 appear to be a marker of breast cancer aggressiveness and in breast cancer may have a pro-angiogenic rather than anti-angiogenic role. PMID- 17912440 TI - Imatinib impairs the proliferation and function of CD4+CD25+ regulatory T cells in a dose-dependent manner. AB - The tyrosine kinase inhibitor imatinib has been reported to inhibit CD8+ T lymphocytes. Little is known about its effects on CD4+CD25+ regulatory T cells (T(reg) cells) which might regulate the graft-vs.-leukemia (GVL) reaction after allogeneic stem cell transplantation (allo-SCT) and donor lymphocyte infusion (DLI). This is of particular interest in patients with relapse of chronic myeloid leukemia (CML) after allo-SCT, as the two therapeutical options DLI and imatinib might interact reversely. Here, we demonstrate that the proliferation of CD4+CD25+ T(reg) cells and their production of IL-10, TGF-beta1 and granzyme B as markers of activation were significantly down-regulated by imatinib in a dose dependent manner. In addition, the expression of surface CD69, both surface and intracellular GITR, FoxP3, CD152 (CTLA) of activated CD4+CD25+ T(reg) cells were inhibited by imatinib in a dose-dependent manner. In light of these findings, clinical administration of imatinib might not result in a reduction of the GVL effect on CML patients receiving imatinib after allo-SCT and/or DLI or other CD8+ T lymphocyte based immunotherapies as the function of CD8+ cytotoxic T lymphocytes and CD4+CD25(hi) Treg cells is hampered in a similar way by imatinib. PMID- 17912441 TI - Enhanced susceptibility to apoptosis of oral squamous cell carcinoma cells subjected to combined treatment with anticancer drugs and phosphatidylinositol 3 kinase inhibitors. AB - The purpose of this study was to determine whether phosphatidylinositol 3-kinase (PI 3-K) inhibitors could modulate the apoptotic activity of the anticancer drugs cisplatin, 5-fluorouracil or docetaxel in an oral squamous cell carcinoma (OSCC) cell line, HSC-2. In preliminary experiments, cisplatin, 5-fluorouracil and docetaxel inhibited the proliferation of OSCC cells in a dose-dependent manner. We found that two PI 3-K inhibitors, wortmannin and LY294002, markedly suppressed the phosphorylation of Akt in OSCC cells. Treatment of OSCC cells with PI 3-K inhibitors significantly enhanced cisplatin-, 5-fluorouracil- or docetaxel induced apoptosis. Caspase-3 and -9 inhibitors, but not a caspase-8 inhibitor, reduced anticancer drug-mediated apoptosis in PI 3-K inhibitor-treated OSCC cells, suggesting that the apoptotic pathway induced by the combination of anticancer drug therapy and PI 3-K inhibition may be functionally related to the intrinsic apoptotic pathway in OSCC cells. Expression of Bcl-2, cellular inhibitor of apoptosis protein-1 (cIAP-1), and X-linked IAP was down-regulated, and expression of Bax was up-regulated by PI 3-K inhibitors, while that of Bcl xL, Bak and cIAP-2 was not attenuated. We also found that Bad phosphorylation was down-regulated by PI 3-K inhibitors. These results suggested that inhibition of PI 3-K enhances the susceptibility of OSCC cells to anticancer drug-mediated apoptosis through regulation of expression and post-translational modification of both pro- and anti-apoptotic proteins. These findings could potentially lead to new strategies for improving the efficacy of anticancer drugs in OSCC cells. PMID- 17912442 TI - Chemopreventive effects of Panax notoginseng and its major constituents on SW480 human colorectal cancer cells. AB - In this study, we evaluated the effects of Panax notoginseng root extract (NGRE) and its major constituents on SW480 human colorectal cancer cells. We used high performance liquid chromatography to determine the contents of major saponins in NGRE. The anti-proliferative effects were evaluated by the cell counting method, and concentration-related anti-proliferative effects were observed. At 1.0 mg/ml, NGRE inhibited cell growth by 85.8% (P<0.01), probably linked to the higher concentration of ginsenosides Rb1 and Rg1. The pharmacologic activities of notoginsenoside R1 and ginsenosides Rg1 and Rb1 on the cells were antiproliferative. We tested the effects of NGRE on DNA synthesis by measuring [3H]-thymidine incorporation. NGRE induced cell apoptosis at 0.5 and 1 mg/ml. Two day treatment with 300 microM of notoginsenoside R1, ginsenosides Rg1 and Rb1 increased cell apoptosis significantly. Cell cycle and cyclin A assay showed that NGRE arrested cells in the synthesis phase and increased the expression of cyclin A remarkably. NGRE also enhanced the actions of two chemotherapeutic agents, 5 fluorouracil and irinotecan. Cell growth decreased more with the combined treatment of NGRE and 5-fluorouracil (or irinotecan) than with the chemotherapy agent applied alone, suggesting that notoginseng can reduce the dose of 5 fluorouracil (or irinotecan) needed to achieve desired effects. Further in vivo and human trials are warranted to test whether notoginseng is a valuable chemo adjuvant with clinical validity. PMID- 17912443 TI - PTEN inactivation in lung cancer cells and the effect of its recovery on treatment with epidermal growth factor receptor tyrosine kinase inhibitors. AB - To understand the mechanisms of PTEN inactivation, which is reported to be involved in tumor progression and drug resistance in lung cancer, we analyzed the expression levels of PTEN at mRNA and protein levels, along with the genetic and epigenetic status of the PTEN gene, in a panel of lung cancer cell lines. Western blot analysis showed that six out of 25 (24%) cell lines displayed low expression of PTEN protein. The level of PTEN mRNA correlated well with corresponding protein expression in each of these six cell lines. In two of the six cell lines genomic analysis revealed homozygous deletions of the PTEN gene. Another two of the six cell lines displayed hypermethylation of the PTEN gene promoter assessed by methylation-specific PCR. The levels of PTEN mRNA and protein expression in PC9/f9 and PC9/f14 cells, which are gefitinib-resistant derivatives of the gefitinib-sensitive cell line, PC9, were reduced compared to the parental line. After treatment with the demethylating agent 5-aza-2'deoxycytidine (5-AZA) and the histone deacetyltransferase (HDAC) inhibitor Trichostatin A (TSA), the expression levels of PTEN mRNA and protein in these four cell lines (PC9/f9, PC9/f14, PC10 and PC14) were actually restored. In summary, reduction in PTEN protein expression was regulated by histone deacetylation and hypermethylation of the gene promoter, as well as homozygous deletion. In addition, we demonstrated that the combination treatment of gefitinib and TSA induced significant growth inhibition in gefitinib-resistant PC9/f9 and PC9/f14 cells. These findings suggest that the combination of the epidermal growth factor receptor tyrosine kinase inhibitor gefitinib with the demethylating agent 5-AZA and the HDAC inhibitor TSA may be a useful strategy for the treatment of some lung cancers. PMID- 17912444 TI - The breast of parous women without cancer has a different genomic profile compared to those with cancer. AB - Our studies are aimed at determining whether pregnancy induces a specific genomic signature in the postmenopausal breast that is responsible for the protective effect elicited by this physiological process. For this purpose we designed a study to compare the gene expression profiles in normal breast tissue from parous postmenopausal women with (case) and without (control) breast cancer. We have used breast samples from 18 parous controls and 41 parous cases. The epithelium and the interlobular stroma were dissected using laser capture microdissection and the RNA of each compartment and each sample was isolated, amplified using PCR methodology, and hybridized to cDNA glass-microarrays containing 40,000 genes, placing the human reference RNA in the green channel (Cy3) and the breast tissue samples in the red channel (Cy5). The normalization and statistical analysis of the expression data were carried out by using the LIMMA software package for the R programming environment which provides functions to summarize the results using the linear model perform hypothesis tests and adjust the p-values for multiple testing. We were able to identify 126 genes that were upregulated and 103 that were downregulated in the parous control group. There were only 56 genes differentially expressed in the interlobular stroma in the parous control group in relation to the other group of women under study. The gene categories that were overrepresented in the breast epithelium of the parous control breast are related to apoptosis, DNA repair, response to exogenous agents and transcription regulation. In the present study we demonstrate that full-term pregnancy imprints a specific genomic signature in the breast epithelium of postmenopausal parous control women that is significantly different from women who have developed cancer. This genomic signature induced by pregnancy could help to predict in which women parity is protective. PMID- 17912445 TI - An oncolytic adenoviral vector carrying the tyrosinase promoter for glioma gene therapy. AB - Targeting gene expression to cancer cells remains a challenge for the development of gene and viral therapy for gliomas. Recent studies have highlighted transcriptional targeting as one of the possible solutions to overcome this limitation. In this context, melanoma associated antigens (MAAs) are usually over expressed in brain tumors in comparison to normal brain tissue. For this reason, we investigated the use of the tyrosinase promoter as a transcriptional element to target oncolytic therapy for gliomas. Tyrosinase mRNA expression was evaluated by qRT-PCR in normal human brain tissue as well as in human glioma specimens. We found that this gene was significantly over-expressed in glioma cell lines and in primary glioma samples. Tyrosinase expression correlated with the grade of the tumor (p-value range: 0.05-0.001). Furthermore, transfection of several cell cultures with human and mouse tyrosinase promoters driving a luciferase reporter gene confirmed the activity of this promoter in mouse and human cells. To evaluate whether tyrosinase-activated conditionally replicative adenoviruses (CRAds) could induce toxicity in glioma cells, two vectors (Ad h/m and Ad24TYR) were tested in a mouse glioma model. C57BL/6 mice underwent intracranial injection of tumor cell line GL261. Survival was used to evaluate efficacy of the tested vectors. Mice receiving 1 x 10(9) MOI of Ad h/m and Ad24TYR following intracranial tumor implants had a median survival of 46+/-3 days (p<0.05); in contrast, those treated with medium had a median survival of 31+/-2 days. These results suggest that injection of tyrosinase CRAds leads to prolongation of survival in mice with experimental brain tumors. The tyrosinase promoter stands as a proof of principle of the potential use of MAA over-expression patterns for targeting novel anti-glioma therapies. PMID- 17912446 TI - Effects of alpha-difluoromethylornithine on thrombospondin-1 production by human breast cancer cells. AB - We have previously observed that inhibition of polyamine biosynthesis with alpha difluoromethylornithine (DFMO) upregulates production of thrombospondin-1 (TSP 1), an extracellular matrix protein with potent anti-angiogenic and antimetastatic properties, by MDA-MB-435 human breast cancer cells in culture. The present experiments were designed to investigate the mechanisms by which DFMO regulates TSP-1 production in this system. 35S-methionine pulse chase experiments indicated that DFMO administration increased TSP-1 synthesis by approximately 6 fold, while it slightly but significantly decreased protein half-life from 35 to 28 min. DFMO treatment increased steady state TSP-1 mRNA levels by 2-fold in MDA MB-435 cells. TSP-1 promoter reporter studies indicated that this increase was largely due to activation of transcription. Analysis of distribution of TSP-1 mRNA levels between non-polysomal, subpolysomal and polysomal fractions in control and DFMO-treated cells suggested a major stimulatory effect of the drug on TSP-1 translation. A similar increase in TSP-1 transcription and translation in response to DFMO treatment was also observed in vivo in MDA-MB-435 breast cancer xenografts. Surprisingly however, we failed to detect an increase in TSP-1 protein as assessed by Western blot analysis. The reason for this unexpected finding is unknown but may be due to DFMO-induced stimulation of TSP-1 secretion into the systemic circulation, thus preventing its accumulation within the tumor. PMID- 17912447 TI - Glioblastoma multiforme cells: expression of erythropoietin receptor and response to erythropoietin. AB - Erythropoietin (EPO) is a glycoprotein hormone that is a primary regulator of erythropoiesis. In erythroid cells, EPO binds to its receptor (EPOR) to stimulate growth, prevent apoptosis, and promote differentiation. Both EPO and EPOR have been found in many normal and tumor nonerythroid cell types. EPO has been reported to stimulate proliferation and inhibit apoptosis of cancer cells. In this study, we found that EPOR is expressed in brain tumors, glioma cell lines and explants, as well as, normal brain. EPO slightly stimulated the growth of serum-starved glioma cells. Furthermore, EPO increased the phosphorylation of AKT through the PI3K pathway in the glioma cells. It also increased the phosphorylation of ERK, c-jun, JNK, as well as, the expression of BCL-2 and BCL xl in these cells. These results suggest that the EPO-EPOR pathway may promote glioma cell survival and could become a therapeutic target in brain tumors. PMID- 17912448 TI - Interaction of prostate specific membrane antigen with clathrin and the adaptor protein complex-2. AB - Prostate-specific membrane antigen (PSMA) is an integral membrane glycoprotein expressed in prostatic epithelia and is being evaluated as a therapeutic target in prostate cancer. It undergoes constitutive receptor-mediated endocytosis via clathrin-coated pits, which is enhanced in the presence of monoclonal antibodies directed against it. We describe distinct interactions of PSMA with clathrin and the clathrin adaptor protein-2 (AP-2) complex, two components of clathrin-coated pits. The intracellular N-terminal domain of PSMA interacts with the N-terminal globular domain of clathrin heavy chain. Deletion analysis revealed an important determinant of this interaction residing within the proximal portion of the clathrin heavy chain N-terminal domain (amino acids 1-85) distinct from the clathrin binding sites of other known clathrin-binding proteins. Furthermore, PSMA interacts with the ear domain of alpha-adaptin (an AP-2 subunit), and a glutamic acid residue at position 7 in the cytoplasmic tail of PSMA is essential for this interaction. These data indicate that PSMA exhibits a high affinity, specific association with the clathrin-based endocytic machinery by distinct interactions with both clathrin and AP-2. Thus, although PSMA is a new member of the dual AP and clathrin binding proteins, its alpha-adaptin and clathrin heavy chain binding determinants are distinct from those of other members. PMID- 17912449 TI - MAGED2: a novel p53-dissociator. AB - The tumor suppressor protein p53 is a transcription factor that is frequently mutated in human cancers. In response to DNA damage, unmutated or wild-type (wt) p53 protein is stabilized and activated by post-transcriptional modifications that enable it to induce either apoptosis or cell cycle arrest. Using a yeast p53 dissociator assay, we identified MAGED2 as a potential negative regulator of wt p53 activity. Subsequently, using co-immunoprecipitation and reporter gene assays in human cultured cells that are often adopted for functional analysis of p53 we demonstrated that MAGED2 interacted physically with p53 and modified its activity. Finally, we were able to illustrate expression of both p53 and MAGED2 within the same subcellular compartment, i.e. either nucleus or cytoplasm, in 2,682 human cancer tissue specimens using a common cancer tissue microarray and antibodies against MAGED2 and p53. The present results implicate MAGED2, a novel protein, as a p53-dissociator. PMID- 17912450 TI - Targeting of a conditionally replicative adenovirus agent to human squamous cell carcinomas of the head and neck. AB - Conventional cancer treatments are not adequate for the majority of most patients stricken with squamous cell carcinomas of the head and neck (SCCHN). Conditionally replicating adenoviruses (CRAds) represent a promising new modality for treating of neoplastic diseases, including SCCHN. Specifically, CRAd agents infect tumor cells and selectively replicate within them, thus causing their death while sparing surrounding normal cells in the host. Oncolysis results from the replicative life cycle of the virus, which lyses infected tumor cells and releases viral progeny for propagation of infection and resultant lysis of neighboring cancer cells, sparing normal host cells. However, to date there have been two main limitations to successful clinical application of these CRAd agents: poor infectivity and poor tumor specificity. Here we report the construction of a CRAd agent, CRAd-CXCR4.F5/3, in which the adenovirus E1 gene is driven by a tumor-specific CXCR4 promoter, and the viral infectivity is enhanced by a fiber modification, F5/3, containing an Ad3 knob chimeric fiber protein. As expected, this agent improved both of the viral infectivity and tumor specificity as evaluated in established SCCHN tumor cell lines and in primary tumor tissues from multiple patients. As an added benefit, the activity of the CXCR4 promoter was low in human liver as described previously. Based on these data, the CRAd CXCR4.F5/3 is a promising novel CRAd agent for SCCHN targeting with low host toxicity. PMID- 17912451 TI - Transactivation of HER2 by vasoactive intestinal peptide in experimental prostate cancer: Antagonistic action of an analog of growth-hormone-releasing hormone. AB - Receptors for vasoactive intestinal peptide (VIP) and the human epidermal growth factor family of tyrosine kinase receptors (HER) are potent promoters of cell proliferation, survival, migration, adhesion and differentiation in prostate cancer cell lines. In this study, we analyzed the cross-talk between both classes of receptors through the regulation of HER2 transactivation and expression by VIP. Three growth-hormone-releasing hormone analogs endowed with antagonistic activity for VIP receptors (JV-1-51, -52, and -53) abrogated the autocrine/paracrine stimuli of VIP on androgen-independent PC3 cells in the absence or the presence of 10% fetal bovine serum. Semiquantitative and real-time quantitative RT-PCR together with Western blotting showed increased expression levels of both mRNA and proteins for HER2 and HER3 in PC3 and androgen-dependent LNCaP prostate cancer cells as compared to non-neoplastic RWPE-1 cells. VIP (100 nM) stimulated the expression levels of both HER2 and HER3 in PC3 cells in a time dependent manner. Whereas these effects were relatively slow, VIP rapidly (0.5 min) increased HER2 tyrosine phosphorylation. This pattern of HER transactivation was blocked by H89, a protein kinase A (PKA) inhibitor, as well as by the specific VIP antagonist JV-1-53, indicating the involvement of VIP receptors and PKA activity in phosphorylated HER2 formation. These findings support the merit of further studies on the potential usefulness of VIP receptor antagonists and both HER2 antibodies and tyrosine kinase inhibitors for prostate cancer therapy. PMID- 17912452 TI - Novel substituted 1,4-anthracenediones with antitumor activity directly induce permeability transition in isolated mitochondria. AB - Synthetic analogs of 1,4-anthraquinone (AQ code number), which block nucleoside transport, decrease DNA, RNA and protein syntheses, trigger cytochrome c release without caspase activation, induce apoptotic DNA fragmentation and inhibit the proliferation of wild-type and multidrug resistant tumor cells in the nM range in vitro, rapidly cause the collapse of mitochondrial transmembrane potential in cell and cell-free systems. Because mitochondrial permeability transition (MPT) requires more than depolarization to occur, antitumor AQs were tested for their ability to directly trigger specific markers of MPT in isolated mitochondria. In contrast to a spectrum of conventional anticancer drugs that are inactive, various AQs interact with isolated mitochondria in a concentration- and time dependent manner to rapidly cause large amplitude swelling and Ca2+ release in relation with their effectiveness against L1210, HL-60 and LL/2 tumor cells in vitro. Indeed, the lead antitumor AQ8, AQ9 and AQ17 are also the most effective inducers of MPT in isolated mitochondria, whereas all AQ derivatives devoid of anti-proliferative activity also fail to trigger mitochondrial swelling and Ca2+ release. Moreover, the ability of 4 microM AQ17 to maximally induce mitochondrial swelling and Ca2+ release within 15 min is similar to that of classic MPT inducing agents, such as 5 microg/ml alamethicin, 200 microM atractyloside, 5 microM phenylarsine oxide, 100 microM arsenic trioxide and a 100 microM Ca2+ overload. Interestingly, AQ17 requires a priming concentration of 20 microM Ca2+ to trigger mitochondrial swelling and Ca2+ release and these 0.1 microM ruthenium red-sensitive MPT events are abolished by 1 microM cyclosporin A, 2 mM ADP and 20 microM bongkrekic acid, which block components of the permeability transition pore (PTP), and also inhibited by 50-100 microM of various ubiquinones, which interact with the quinone binding site of the PTP and raise the Ca2+ load required for PTP opening. Hence, antitumor AQs that target isolated mitochondria and trigger MPT might directly interact with components of the PTP to induce conformational changes that increase its Ca2+ sensitivity and transition from the closed to the open state. PMID- 17912453 TI - Liposome-incorporated Grb2 antisense oligodeoxynucleotide increases the survival of mice bearing bcr-abl-positive leukemia xenografts. AB - We previously demonstrated that liposome-incorporated antisense oligodeoxynucleotide specific for the grb2 mRNA (L-Grb2) inhibited Grb2 protein expression and the proliferation of bcr-abl-positive leukemia cell lines. To determine whether L-Grb2 has the potential of being a therapeutic modality against bcr-abl-positive leukemia, we studied the tissue distribution of L-Grb2 in normal mice before studying its effects in mice bearing bcr-abl-positive leukemia xenografts. L-Grb2 was widely distributed in the body. The highest tissue concentrations of L-Grb2 were found in the spleen and liver, which are the organs where the tumor mass of bcr-abl-positive leukemia is mainly found. At 4 h post-injection, the amount of L-Grb2 detected per g of tissue was 64 microg in spleen and 50 microg in liver. Intravenous injection of bcr-abl-positive 32D mouse leukemia cells into radiated NOD/scid mice caused a lethal leukemia syndrome; we determined whether L-Grb2 could prolong the survival of mice bearing such xenografts. One day after leukemia cell inoculation, mice received twice weekly intravenous injections of L-Grb2. At an injection dose of 15 mg of L-Grb2 per kg of mouse body weight, 80% of mice treated with L-Grb2 survived to 48 days (end of study) whereas 0% of mice treated with the same dose of liposomal control oligonucleotide survived; the mean survival duration of these groups was 44 and 20 days, respectively. Our data indicate that L-Grb2 prolonged the survival of mice bearing bcr-abl-positive leukemia xenografts. L-Grb2 may be used as a novel cancer therapeutic modality. PMID- 17912454 TI - The regulatory beta-subunit of protein kinase CK2 accelerates the degradation of CDC25A phosphatase through the checkpoint kinase Chk1. AB - Human CDC25 phosphatases play an important role in cell cycle regulation by removing inhibitory phosphate groups on cyclin-CDKs. Chk1 has been shown to phosphorylate CDC25 family members down-regulating their phosphatase activity through distinct mechanisms. The kinase activity of Chk1 is evident in unperturbed cells and becomes enhanced in response to DNA damage or stalled replication. We have previously shown that the activity of Chk1 is increased following interaction with the regulatory beta-subunit of protein kinase CK2. In the present study, ectopic expression of CK2beta during normal cell cycle progression is shown to enhance CDC25A degradation, and this occurs in a manner similar to that by which CDC25A is down-regulated upon activation of cellular checkpoint responses. By using RNA interference to specifically deplete cells of Chk1, we demonstrate that Chk1 mediates the down-regulation of endogenous CDC25A, which occurs upon induction of CK2beta expression. When degradation of CDC25A is induced by CK2beta during activation of the G2 checkpoint, it leads to partial dephosphorylation of Cdc2 at its inhibitory residue Tyr15. These results suggest that protein kinase CK2 is involved in cell cycle regulation and indicate the mechanism by which CDC25A turnover might be regulated by Chk1 in the absence of DNA damage. PMID- 17912455 TI - N-hydroxy-N'-(3,4,5-trimethoxyphenyl)-3,4,5-trimethoxy-benzamidine, a novel resveratrol analog, inhibits ribonucleotide reductase in HL-60 human promyelocytic leukemia cells: synergistic antitumor activity with arabinofuranosylcytosine. AB - Resveratrol (3,4',5,-trihydroxystilbene, RV), an ingredient of wine, exhibits a broad spectrum of antiproliferative effects against human cancer cells. In order to develop a derivative with comparable effects, we modified the molecule by introducing additional methoxyl groups. The resulting novel RV analog, N-hydroxy N'-(3,4,5-trimethoxyphenyl)-3,4,5-trimethoxybenzamidine (KITC), was investigated in HL-60 human promyelocytic leukemia cells. The induction of apoptosis was determined employing a specific Hoechst/propidium iodide double staining method and cell cycle distribution was evaluated by FACS. KITC's influence on the concentration of deoxyribonucleoside triphosphates, the products of ribonucleotide reductase (RR), was determined using the HPLC method. In addition, we analyzed the effects of KITC treatment on the incorporation of 14C-cytidine into the DNA of tumor cells in order to quantify the loss of RR in situ activity. To reveal a potential value of KITC for supporting conventional chemotherapy, we also examined whether a combination of KITC with arabinofuranosylcytosine (Ara-C) could yield synergistic growth inhibitory effects. KITC caused a dose-dependent induction of apoptosis, whereas no remarkable changes of the cell cycle distribution were observed. Incubation with KITC resulted in a significant depletion of intracellular dTTP and dATP pools and was also found to remarkably reduce the in situ activity of RR, the key enzyme of de novo DNA synthesis. In addition, KITC exhibited synergistic combination effects when applied sequentially with Ara-C. Due to these promising results, KITC deserves further preclinical and in vivo testing. PMID- 17912456 TI - Transferrin reverses the anti-invasive activity of human prostate cancer cells that overexpress sema3E. AB - In vitro invasion and adhesion of stably semaphorin (sema) 3E-transfected PC-3 prostate cancer cells were determined in the presence and absence of transferrin. Invasion and adhesion decreased compared to untransfected cells; however, transferrin reversed the effects. Transferrin differentially regulated E-cadherin and beta-catenin in these cells. Insulin growth factor 3 (IGFBP3) negated the invasive and adhesive effects of transferrin. Transferrin increased binding of insulin growth factor (IGF)-1 to the activated IGF-1 receptor, and IGF-1 mimicked the invasive and adhesive effects of transferrin. These data suggest that transferrin modulates sema3E-transfected cells through an IGFBP3/IGF-1-dependent pathway, in part, by regulation of adhesion proteins. PMID- 17912457 TI - Tetraspanin CD9 is involved in the migration of retinal microvascular endothelial cells. AB - Members of the tetraspanin protein family are modulators of several fundamental cellular processes in various cell types. However, expression and function of these proteins have not been studied in microvascular endothelial cells despite their (patho-)physiological importance. Western blotting, FACS or RT-PCR analyses confirmed that CD9 and other tetraspanins are expressed in immortalized microvascular endothelial cells of the bovine retina (iBREC). In subconfluent cultures, most of the detected CD9 was located intracellularly as well as in the plasma membrane at cell-cell contact sites and in long spike-like extensions, whereas cells in confluent cultures predominantly showed plasma membrane staining. In wound healing assays, CD9 delocalized from the plasma membrane to its intracellular compartment in cells located at the gap border, and the gap closure was retarded by the addition of an anti-CD9 antibody. Migration of iBREC towards fibronectin and their adhesion to fibronectin were also strongly inhibited in the presence of an anti-CD9 antibody whereas other anti-tetraspanin antibodies had no effect. In summary, iBREC express members of the tetraspanin family of which CD9 was demonstrated to have a function in migration and adhesion of these cells. PMID- 17912458 TI - Mutations in exons 2 and 3 of the FOLR1 gene in demented and non-demented elderly subjects. AB - We have previously reported six novel mutations in the 5'-UTR of the gene for folate receptor-alpha (FOLR1). In our search for additional mutations we screened patients, referred for investigation of suspected dementia (DGM subgroup) by SSCP and DNA sequencing from the end of exon 1 to the first bases of intron 3. We found 4 sequence variations, FOLR1 g.1314G>A, g.1816delC, g.1841G>A, and g.1928C>T. Pyrosequencing genotyping assays were developed for all of them, and 389 active seniors (AS subgroup) and the 202 DGM patients were genotyped for these mutations. The frequency q of the mutated allele was, among the AS subjects, 0.068, 0.0026, 0.0026, and 0.024 respectively, and among the DGM subjects, 0.067, 0.0076, 0.0078, and 0.023. The g.1816delC and g.1841G>A mutations thus were more frequent in the DGM than in the AS subgroup, but the difference did not reach statistical significance. The mutated alleles, FOLR1 1816(-) and 1841A, always occurred together in the same subjects, suggestive of a rare double-mutant haplotype. The two common polymorphisms, FOLR1 g. 1314G>A and g.1928C>T seemed not to raise tHcy plasma levels, whereas the double-mutated g.1816(-)-g.1841A haplotype may possibly have a slight tHcy-raising effect. Thus, so far 8 novel rare FOLR1 mutations with a combined prevalence of approximately 1.3% in Whites as well as two common polymorphisms with 5% and 13%, respectively, have been demonstrated. Only a few of the rare mutations may potentially be associated with raised plasma tHcy concentrations. No association with dementia was found. PMID- 17912459 TI - Induction of apoptosis underlies the Radix Rubiae-mediated anti-proliferative action on human epidermal keratinocytes: implications for psoriasis treatment. AB - Psoriasis is a chronic inflammatory skin disease characterized histologically by hyperproliferation and aberrant differentiation of epidermal keratinocytes. While screening 60 psoriasis-treating Chinese herbs for their anti-proliferative properties using a cultured human HaCaT keratinocyte model, we found Radix Rubiae to be highly effective. Evidence is now provided that induction of apoptosis is the underlying mechanism for the observed anti-proliferative action of Radix Rubiae. Analysis of cell cycle with PI staining showed that Radix Rubiae induced the appearance of a sub-G1 peak and cell arrest at the G1 phase. Radix Rubiae was also capable of inducing morphological changes as evidenced by nuclear condensation. DNA fragmentation was clearly demonstrated by gel electrophoresis and by the TUNEL method. Quantitative analyses by Annexin V-PI staining revealed that Radix Rubiae-induced apoptosis was dose- and time-dependent. Furthermore, Radix Rubiae was able to activate caspase-3 expression when examined by Western blot analysis. The cellular, morphological and molecular data unequivocally demonstrated that induction of cellular apoptosis was mainly responsible for the previously observed anti-proliferation induced Radix Rubiae on HaCaT keratinocytes. Our experimental results suggest that Radix Rubiae is a promising source from which a herb-based topical agent could be developed for psoriasis treatment. PMID- 17912460 TI - Therapeutic immune response induced by intratumoral expression of the fusogenic membrane protein of vesicular stomatitis virus and cytokines encoded by adenoviral vectors. AB - We assessed whether intratumoral expression of the fusogenic membrane protein of vesicular stomatitis virus (VSV-G), encoded by a replication-defective adenovirus vector (Ad.VSV-G), alone or in combination with local coexpression of cytokines induces tumor-specific immune responses in a syngeneic murine colon cancer model. We confirmed in vitro by dye colocalization that transduction of murine cells with Ad.VSV-G induces cell-cell fusion. In a bilateral syngeneic subcutaneous colon cancer model in C57BL/6 and BALB/c mice, we demonstrated that intratumoral injection of Ad.VSV-G leads to a significant growth reduction of the directly vector-treated tumor, but also of the contralateral not directly vector-treated tumor. When compared to monotherapy, the anti-neoplastic efficacy was significantly enhanced when intratumoral Ad.VSV-G administration was combined with adenovirus vectors encoding IL-2, IL-12, IL-18, IL-21, or GM-CSF. The anti tumor effects of the first three cytokines in combination with VSV-G expression were somewhat greater than those of the latter two. However, the differences did not reach statistical significance. The combination therapy resulted also in a significantly enhanced survival when compared to monotherapy. In addition, we demonstrated that intratumoral expression of VSV-G in combination with the tested cytokines induced a strong tumor-specific cytotoxic T lymphocyte (CTL) response and infiltration of tumors with macrophages. The effects of the combination therapy were clearly greater than those of the monotherapy. Our experimental data indicate that intratumoral expression of VSV-G, particularly in combination with cytokines, is a promising novel tool for the development of in situ tumor vaccination approaches. PMID- 17912461 TI - Highly stable plasminogen activator inhibitor type one (VLHL PAI-1) protects fibrin clots from tissue plasminogen activator-mediated fibrinolysis. AB - Plasminogen activator inhibitor-1 (PAI-1) is the major specific inhibitor of tissue-type plasminogen activator (tPA) which mediates fibrin clot lysis through activation of plasminogen. Wild-type-PAI-1 (wPAI-1) is rapidly converted to the latent form (half-life of approximately 2 h) and loses its ability to inhibit tPA. We developed a very long half-life PAI-1 (VLHL PAI-1), a recombinant protein with a half-life >700 h compared with wPAI-1. In this study, VLHL PAI-1 was assessed for its ability to inhibit clot lysis in vitro. Clot formation was initiated in normal plasma supplemented with tPA by the addition of either tissue factor or human recombinant FVIIa. Clot lysis time, monitored turbidimetrically in a microtiter plate reader, was determined at various concentrations of wPAI-1 and VLHL PAI-1. Both wPAI-1 and VLHL PAI-1 caused a significant increase in clot lysis time, although the latter was somewhat less effective at lower concentrations. The VLHL PAI-1, but not wPAI-1, maintained its anti-fibrinolytic activity after preincubation overnight at 37 degrees. These studies demonstrate that VLHL PAI-1 is an effective inhibitor of fibrin clot degradation. Due to the high stability of VLHL PAI-1 compared with wPAI-1, this novel inhibitor of tPA mediated fibrinolysis may have therapeutic applications for treating surgical and trauma patients when used directly or in conjunction with the procoagulant recombinant FVIIa. PMID- 17912462 TI - Genes regulated by interferon-gamma in human uterine microvascular endothelial cells. AB - Interferon (IFN)-gamma plays a critical role in murine uterine spiral artery remodeling for successful pregnancy. The effect of IFN-gamma on human uterine microvasculature, however, remains poorly understood. The aim of this study was to identify the genes regulated by IFN-gamma in human uterine microvascular endothelial cells. The effect of IFN-gamma on the gene expression profile in human uterine microvascular endothelial cells was evaluated by cDNA microarray analysis and quantitative real-time reverse transcriptase-polymerase chain reaction for the selected genes of interest. In vivo expression of the protein encoded by some of these genes in human uterine microvascular endothelial cells was evaluated by Western blotting and immunohistochemistry. Treatment with 10 ng/ml IFN-gamma for 4 h induced a significant > or =2-fold change in 29 genes in pooled human uterine microvascular endothelial cells; a total of 20 genes were up regulated, whereas nine genes were down-regulated. The genes significantly up regulated included chemokines (CXCL9, CXCL10, CCL8, IL15RA, and CCL5), enzymes (GBP5, TAP1, CYP27B1, SOD2, MX1, CASP1, and PTGES), and transcription factors (TFAP2C, IRF1, NFE2L3). The genes significantly down-regulated following IFN gamma treatment included cytokines/cytokine receptors (CSF2, IL1R2, and SPP1), and insulin-like growth factor binding proteins (WISP2 and IGFBP3). The results of the cDNA microarray analysis were confirmed by quantitative real-time reverse transcriptase-polymerase chain reaction for the selected 17 genes of interest. The immunoreactivity for the proteins encoded by IL15RA, IFI30, and MX1 was detected in human uterine microvascular endothelial cells in vivo, whereas the immunoreactivity for CCNA1 and NQO1 was not detectable. These results suggest that IFN-gamma regulates the gene expression involved in natural killer cell recruitment, embryo and trophoblast migration, endometrial decidualization, angiogenesis, angiostasis, and anti-viral infection in human uterine microvascular endothelial cells. PMID- 17912463 TI - Increased expression of erythropoietin receptor in human pterygial tissues. AB - Erythropoietin (Epo) induces physiological activities such as cell proliferation, migration, and angiogenesis in Epo receptor (EpoR)-expressing vascular endothelial and tumor cells. Recently, it has been demonstrated that growth factor-independent proliferation is frequently observed during the cell transformation process. Pterygium is a fibrovascular proliferating tissue that includes transformed cells. The aim of this study was to examine the localization of Epo and EpoR proteins in human pterygial tissues. Eleven samples including nine pterygia and two normal bulbar conjunctivas, which were surgically excised, were studied. Formalin-fixed, paraffin-embedded tissue sections were constructed and then were examined by immunohistochemistry with anti-Epo and EpoR antibodies. Cytoplasmic immunoreactivity for EpoR was heterogeneously detected in basal and suprabasal cells of the pterygium epithelium. In the pterygium stroma, a variety of endothelial cells forming vascular cavities showed cytolasmic immunoreactivity for EpoR. In normal conjunctival epithelium, a few basal cells showed a weak homogeneous immunoreactivity for EpoR in the cytoplasm. The number of EpoR expressing epithelial cells was much higher in the pterygium compared to the normal conjunctiva. EpoR expression was marginally detected in stromal microvessels of the normal conjunctiva. Immunoreactivity for Epo was not noted in pterygium epithelium and stroma, and in normal conjunctiva. These results suggest that the Epo-independent EpoR-signaling pathway plays a potential role in cell proliferation and angiogenesis in human pterygium. PMID- 17912464 TI - Association of a polymorphism of CYP3A4 with type 2 diabetes mellitus. AB - Although several environmental factors, including a high-calorie diet and physical inactivity, influence the development of type 2 diabetes mellitus, genetic factors have been shown to contribute to individual susceptibility to this condition. The purpose of the present study was to identify gene polymorphisms that confer susceptibility or resistance to type 2 diabetes mellitus, and thereby to contribute to assessment of the genetic risk for this condition. The study population comprised 5259 unrelated Japanese individuals (2980 men, 2279 women), including 1640 subjects with type 2 diabetes mellitus (1071 men, 569 women) and 3619 controls (1909 men, 1710 women). The genotypes for 94 polymorphisms of 67 genes were determined with a method that combines the polymerase chain reaction and sequence-specific oligonucleotide probes with suspension array technology. Evaluation of genotype distributions by the chi square test revealed that the 13989-->G (Ile118Val) polymorphism of the cytochrome P450, subfamily IIIA, polypeptide 4 gene (CYP3A4) was significantly (false discovery rate, 0.000009) associated with the prevalence of type 2 diabetes mellitus. Multivariable logistic regression analysis with adjustment for age and sex also revealed that the 13989-->G (Ile118Val) polymorphism of CYP3A4 was significantly (P=0.00002) associated with the prevalence of type 2 diabetes mellitus, with the AG genotype being protective against this condition. Genotyping for CYP3A4 may thus prove informative for assessment of the genetic risk for type 2 diabetes mellitus. PMID- 17912465 TI - Overexpression of regucalcin suppresses gene expression of insulin signaling related proteins in cloned rat hepatoma H4-II-E cells: involvement of insulin resistance. AB - Overexpression of regucalcin has been shown to enhance glucose utilization and lipid production in the cloned rat hepatoma H4-II-E cells in vitro, and it induces insulin resistance. The effect of regucalcin on the gene expression of insulin signaling-related proteins was investigated in the cloned rat hepatoma H4 II-E cells overexpressing regucalcin in vitro. The hepatoma cells (wild-type) and stable regucalcin/ pCXN2-transfected cells (transfectants) were cultured for 72 h in a medium containing 10% fetal bovine serum (FBS) to obtain subconfluent monolayers. Cells with subconfluency were cultured for 24, 48, or 72 h in a medium containing either vehicle or insulin (10(-9)-10(-7) M) with or without supplementation of glucose (10, 25, or 50 mg/ml of medium). The expression of rat insulin receptor (Insr), phosphatidylinositol 3-kinase (PI3K), glucose transporter 2 (GLUT 2), or glyceroaldehyde-3-phosphate dehydrogenase (G3PDH) mRNAs was examined using reverse transcription-polymerase chain reaction analysis with specific primers. GLUT 2 mRNA expression was significantly increased in the transfectants, while Insr, PI3K, and G3PDH mRNA levels were not significantly changed in the transfectants. Culture with insulin (10(-8) or 10(-7) M) caused a significant increase in PI3K mRNA levels in wild-type cells cultured for 24 or 48 h, while it had no effect on Insr mRNA levels. The supplementation of glucose (10, 25, or 50 mg/ml) caused a significant increase in Insr and PI3K mRNA levels in wild-type cells. The effect of insulin or glucose supplementation on these gene expression levels was not seen in the transfectants. The combination of insulin (10(-7) M) and glucose (50 mg/ml) caused a significant increase in Ins and PI3K mRNA levels in wild-type cells. Such an effect was not seen in the transfectants. Culture with insulin or glucose supplementation failed to have a significant effect on GLUT2 and G3PDH mRNA levels in the wild-type cells or transfectants. This study demonstrates that overexpression of regucalcin suppresses the enhancing effect of insulin or glucose on the gene expression of insulin signaling-related proteins in the cloned rat hepatoma H4-II-E cells. PMID- 17912466 TI - Genetic polymorphisms of molecules associated with inflammation and immune response in Japanese subjects with functional dyspepsia. AB - Inflammatory changes in the gastric mucosa are commonly observed in Japanese patients with functional dyspepsia (FD). However, detailed data regarding the relationship between the genetic regulatory factors of inflammation and FD are not available. We investigated the associations between FD and genetic polymorphisms of molecules associated with inflammation or immune response (IL 17A, -17F and MIF). The study was performed with 278 subjects (188 with no upper abdominal symptoms and 90 with FD according to the Roma III criteria). We employed the PCR-SSCP (multiplex PCR for IL-17A and -17F) method to detect the gene polymorphisms. Overall, the polymorphisms of the IL-17A, -17F and MIF genes were not correlated with the susceptibility to FD. However, the MIF -173C allele carrier had a significantly increased risk for the development of epigastric pain syndrome (EPS) of FD (OR, 2.12; 95% CI, 1.00-4.49; p=0.0497). In Helicobacter pylori (H. pylori)-infected cases, the number of IL-17F 7488T alleles was positively correlated with the development of EPS (OR, 11.3; 95% CI, 1.23-103.2; p=0.032), while the IL-17F T/T homozygote and the MIF -173C carrier had an increased risk for EPS (OR, 10.4; 95% CI, 1.17-92.3; p=0.036 and OR, 3.66; 95% CI, 1.19-11.3; p=0.024, respectively). In addition, a significant interaction between the IL-17F 7488 polymorphism and H. pylori infection was shown to increase the activity and inflammation scores (p=0.043 and 0.042, respectively). There were no significant associations between the IL-17A polymorphism and FD. Our results provide the first evidence that the IL-17F and MIF gene polymorphisms are significantly associated with the development of FD, particularly EPS, a subgroup of FD, in H. pylori-infected subjects. The genetic polymorphisms of inflammation or immune response-related molecules are involved in the development of one of the FD subgroups via H. pylori-induced gastric inflammation. PMID- 17912467 TI - Crosstalk between high-molecular-weight adiponectin and T-cadherin during liver fibrosis development in rats. AB - Adiponectin, a circulating adipocyte-derived secretory protein, reportedly plays an important role in liver fibrosis development, although the biological role of adiponectin in liver fibrogenesis is still controversial. Adiponectin is present in the serum as three oligometric complexes; namely, high-, middle-, and low molecular weight (HMW, MMW, and LMW, respectively). Adiponectin exerts different biological activities in an oligomerization-dependent manner. The aim of our current study was to examine the alteration of each isoform of adiponectin and its receptors (AdipoR1, AdipoR2, and T-cadherin) during the choline-deficient L amino acid-defined (CDAA) diet-induced rat liver fibrosis development. We also elucidated the methylation status of all receptors. The serum level of total adiponectin significantly increased during the liver fibrosis development. Among the three isoforms, only HMW adiponectin was significantly up-regulated whereas MMW and LMW were not. The expression of T-cadherin, which exclusively binds with HMW adiponectin, was significantly augmented as well. The AdipoR2 expression was markedly decreased and showed no marked difference from that of AdipoR1. No obvious methylation change was observed in all three receptors, suggesting that another mechanism is involved in the alteration of receptor gene expression. Collectively, since the specific ligand and receptor were augmented together, crosstalk between HMW adiponectin and T-cadherin may play an important role during liver fibrosis development in rats. PMID- 17912468 TI - Is the p53 codon 72 polymorphism a key biomarker for cervical cancer development? A meta-analysis review within European populations. AB - Human papillomavirus (HPV) is the necessary cause for cervical cancer development, and the interaction of HPV-E6 with p53 is known as the most important event in HPV-associated carcinogenesis. In vitro studies have suggested that HPV-E6 interacts more efficiently with the arginine (Arg) p53 variant at position 72 as it appears to be more susceptible to degradation through the ubiquitin proteasome pathway. However, few reports have corroborated this data, and the role of the p53 codon 72 polymorphism in the development of cervical cancer requires further elucidation. We performed a meta-analysis review of all studies published within European populations to summarize the overall risk of this polymorphism considering the influence of the geographical/ethnic location as an important factor in defining a genetic profile and the susceptibility for cervical cancer development. Our analysis revealed that the p53 Arg/Arg genotype does not seem to represent a risk marker for the development of cervical lesions in the majority of the European countries analysed. However, in countries with low incidence rates of cervical cancer, this polymorphism might represent a significant genetic marker. PMID- 17912469 TI - DNA repair gene XRCC1 and XPD polymorphisms and the risk of idiopathic azoospermia in a Chinese population. AB - Genetic polymorphisms in DNA repair genes may influence individual variation in DNA repair capacity and further influence the risk of developing cancer. However, little information is available on these polymorphisms in infertility. To investigate whether polymorphisms in DNA repair genes, X-ray repair cross complementing group 1 (XRCC1) and xeroderma pigmentosum group D (XPD), alone or in combination, are associated with the risk of developing idiopathic azoospermia, the genotype and allele frequencies of three observed polymorphisms (XRCC1 Arg194Trp and Arg399Gln, and XPD Lys751Gln) were examined by polymerase chain reaction-restriction fragment length polymorphism based on a Chinese population consisting of 171 idiopathic azoospermia patients and 247 normal spermatogenesis fertile controls. Associations between the polymorphisms and the idiopathic azoospermia risk were estimated by logistic regression, and the Statistical analysis system was used to test the gene-gene joint effects. All observed polymorphisms were in agreement with Hardy-Weinberg equilibrium. The XPD 751Gln allele seemed to be a risk allele for azoospermia, with a frequency of 11.40% in the cases and 5.67% in the controls (p=0.004). Compared with the Lys/Lys genotype, the XPD 751 Lys/ increased 5.100- or 3.064-fold, respectively, when combined with the XRCC1 194 Arg/Arg or 399 Arg/Arg genotype. In conclusion, our study provided the first evidence that the XPD and XRCC1 polymorphisms contributed to the risk of developing idiopathic azoospermia in a selected Chinese population. PMID- 17912470 TI - Oxidant-induced cardiomyocyte injury: identification of the cytoprotective effect of a dopamine 1 receptor agonist using a cell-based high-throughput assay. AB - Myocyte injury due to myocardial reperfusion injury plays a crucial role in the pathogenesis of acute myocardial infarction even after successful coronary revascularization. Identification of compounds that reduce reperfusion-associated myocyte death is important. Therefore, we developed an in vitro model of myocardial reperfusion injury in H9c2 rat cardiomyocytes and applied a cell-based high-throughput approach to screen a standard library of pharmacologically active compounds (LOPAC) in order to identify drugs with cardioprotective effects. Oxidative stress was induced with hydrogen peroxide (H2O2) treatment, which resulted in approximately 50% reduction in cell viability. Test compounds were added at a 3-microM final concentration as a pretreatment or in a delayed fashion (30 min after the peroxide challenge in order to imitate pharmacological treatment following angioplasty). Cells were cultured for 3 or 24 h. Viability was quantitated with the methylthiazolyldiphenyl-tetrazolium bromide method. Cytotoxicity and cytoprotection were also evaluated by measuring the lactate dehydrogenase activity in the cell culture supernatant. The screening identified a number of compounds with cytoprotective action, including molecules that are known to interfere with components of DNA repair and cell cycle progression, e.g. poly(ADP-ribose) polymerase (PARP) inhibitors, topoisomerase inhibitors, and cyclin dependent kinase inhibitors, or reduce energy consumption by interfering with cardiac myofilament function. A number of dopamine D1 receptor agonists also provided significant cytoprotection at 3 h, but only three of them showed a similar effect at 24 h: chloro- and bromo-APB and chloro-PB hydrobromide. Chloro APB hydrobromide significantly reduced peroxide-induced PARP activation in the myocytes independently of its action on dopamine D1 receptors, but lacked PARP inhibitor capacity in a cell-free PARP assay system. In conclusion, the pattern of cytoprotective drugs identified in the current assay supports the overall validity of our model system. The findings demonstrate that cytoprotective agents, including novel indirect inhibitors of cellular PARP activation can be identified with the method, chloro-APB hydrobromide being one such compound. The current experimental setting can be employed for cell-based high-throughput screening of various compound libraries. PMID- 17912471 TI - Effect of Angelica gigas extract on melanogenesis in B16 melanoma cells. AB - During the screening of herbs for inhibition of melanogenesis, it was observed that ethanolic extract of Angelicae Gigantis Radix (AGE) effectively inhibited isobutylmethylxanthine-induced melanogenesis in B16 melanoma cells. The melanin content was significantly decreased by AGE in a dose-dependent manner, and no cytotoxicity was observed at the effective concentrations. Decreased melanin content was accompanied by reduced enzyme activity as well as reduced expression of tyrosinase protein and mRNA. The level of tyrosinase-related protein 1 and 2 mRNAs was also decreased by AGE. Additionally, AGE effectively inhibited alpha melanocyte stimulating hormone- and forskolin-induced melanogenesis, and downregulated the mRNA expression of microphthalmia-associated transcription factor, a master transcriptional regulator of melanogenic genes. These results suggest that AGE acts as a putative hypopigmenting agent through downregulation of tyrosinase expression induced via a cAMP-dependent pathway. PMID- 17912472 TI - Radix asari extract protects pancreatic beta cells against cytokine-induced toxicity: implication of the NF-kappaB-iNOS signaling cascade. AB - In this study, we assessed the preventive effects of Radix asari extract (RAE) against cytokine-induced beta-cell destruction. Cytokines secreted by immune cells that have infiltrated pancreatic islets are crucial mediators of beta-cell destruction in insulin-dependent diabetes mellitus. Treatment of RINm5F (RIN) cells with interleukin (IL)-1beta and interferon (IFN)-gamma resulted in a reduction of cell viability and proliferation. However, treatment of RIN cells with RAE protected the IL-1beta and IFN-gamma- mediated viability and proliferation reduction in a concentration-dependent manner. Incubation with RAE also resulted in significant suppression of IL-1beta and IFN-gamma-induced nitric oxide (NO) production, and this reduction was correlated with reduced levels of mRNA and protein associated with the inducible form of NO synthase (iNOS). The molecular mechanism by which RAE inhibited iNOS gene expression appeared to involve the inhibition of NF-kappaB activation as a result of RAE's suppression of IL-1beta and IFN-gamma-induced IkappaBalpha degradation. The protective effects of RAE were verified via the observation of reduced NO generation and iNOS expression, as well as the observation of normal insulin-secretion responses to glucose in IL-1beta and IFN-gamma-treated rat islets. These results suggest that RAE protects beta cells from cytokine toxicity by suppression of NF-kappaB activation. PMID- 17912473 TI - The role of smooth muscle in the pathogenesis of pelvic organ prolapse--an immunohistochemical and morphometric analysis of the cervical third of the uterosacral ligament. AB - The uterosacral ligament (USL) is part of the pelvic floor connecting the uterus to the dorsal body wall and stabilizing it. In samples obtained from hysterectomy patients, the smooth muscle component of the cervical third of the USL was studied by smooth muscle actin (SMA) immunohistochemistry as indicator for structural abnormalities and by morphometric determination of nuclear size as measure for the functional state. From a total of 40 patients, 18 were matched according to age, parity, and premenopausal status. Immunohistochemistry showed a granular patchy distribution of SMA in the pelvic organ prolapse (POP) group. In patients with POP, the size of smooth muscle cell nuclei was significantly reduced in comparison to patients without POP (25.45+/-1.92 microm(2) vs 28.87+/ 2.92 microm(2), p<0.05). We conclude that the smooth muscle component of the USL is impaired in POP patients, indicating a possible role of smooth muscle in the pathogenesis of POP. PMID- 17912475 TI - Recurrent symptomatic chylopericardium after cardiac surgery in a child. AB - Chylopericardium is a rare complication after cardiac surgery in children. We report a case of a recurrent chylopericardium complicating postoperative course in a 4-month-old child. Chylopericardium occurred after two separate operations 6 months apart. On both occasions, symptomatic cardiac tamponade required pericardial drainage. Differentiation of chylopericardium from simple postoperative pericardial effusion can be difficult. When chylopericardium is strongly suspected from previous history and patient background, prompt treatment, including pericardial drainage and dietary treatment, is warranted. PMID- 17912474 TI - Evolution of suiform aromatases: ancestral duplication with conservation of tissue-specific expression in the collared peccary (Pecari tayassu). AB - Aromatase cytochrome P450 (P450arom), the enzyme that catalyzes estrogen synthesis, is required for successful reproduction and is encoded by a single copy gene (CYP19) in most mammals. However, pigs and their distant suiform relatives the peccaries experienced CYP19 duplication. Here, the evolutionary origin of CYP19 duplication, and the evolution of the gene paralogs, was explored further in collared peccaries (Pecari tayassu). Exons IV and V, and the intervening intron, representing duplicated CYP19 genes, were cloned and sequenced from collared peccary, pig, and hippopotamus. Sequence alignment and analysis identified a gene conversion in collared peccary with a breakpoint 102 base pairs (bp) upstream of exon V. Phylogenetic analyses of nucleotide and amino acid sequence upstream of the breakpoint supported a tree in which one peccary sequence was orthologous with the porcine gonadal gene. Cloning and sequencing of tissue transcripts, using reverse-transcriptase polymerase chain reaction techniques (RT-PCR), confirmed that the gonadal ortholog was expressed in collared peccary testis. Orthology of the other genomic sequence with the porcine placental gene was not resolved, but its placenta-specific expression in collared peccary was confirmed by similar transcript analysis. Immunoblot and enzyme activity in collared peccary testes demonstrated much lower levels of P450arom than in pig testis. Collared peccary placental P450arom expression also seemed much lower than pigs. Thus, suiform CYP19 genes arose from an ancestral duplication that has maintained gonad- and placenta-specific expression, but at lower levels in peccaries than pigs, perhaps facilitating the emergence of different reproductive strategies as Suiformes diverged and evolved. PMID- 17912476 TI - Neonatal left pulmonary artery occlusion and postinfarction cysts of the left lung: cause and effect? PMID- 17912477 TI - Double discordance in the hypoplastic left heart. AB - A neonate was transferred for an intracardiac mass. Initially, the mass was mistaken for a hypoplastic right ventricle, a thrombus, or a tumor. Only a thorough segmental and sequential analysis showed double discordance with a hypoplastic left ventricle. No such entity has yet been described. PMID- 17912478 TI - Transseptal left heart catheterization for a patient with a prosthetic mitral valve using live three-dimensional transesophageal echocardiography. AB - The images and videos presented in this article illustrate a diagnostic transseptal left heart catheterization for a patient with a prosthetic mitral valve guided by live three-dimensional (3D) transesophageal echocardiography. This method provided high-quality 3D imaging that was useful in guiding transseptal puncture and demonstrating prosthetic valve function during this evaluation. PMID- 17912479 TI - Cardiomyopathy in multiple Acyl-CoA dehydrogenase deficiency: a clinico pathological correlation and review of literature. AB - Multiple acyl-CoA dehydrogenase deficiency (MADD) is a rare autosomal recessive defect of the electron transfer flavoprotein or ubiquinone oxidoreductase, resulting in abnormal fatty acid, amino acid, and choline metabolism, leading to metabolic acidosis, hypoglycemia, "sweaty-feet" odor, and early neonatal deaths. This report presents a child diagnosed with this disease at birth by newborn screening using the mass spectrometer, who died suddenly at the age of 6 months. The echocardiogram revealed pericardial effusion, thickened ventricular musculature, and insufficiency of both the atrio-ventricular valves. The autopsy showed immense cardiomegaly, fatty infiltration, and hypertrophy of the ventricles. This is the first detailed case report of clinico-pathological correlation of MADD in an infant and brings into light a rare form of cardiomyopathy as a differential diagnosis in critically ill patients. PMID- 17912481 TI - Application of a complication screening method to congenital heart surgery admissions: a preliminary report. AB - There have been comprehensive screening methods developed to identify unwanted inpatient events. A comprehensive assessment of complication diagnoses during congenital heart surgery admissions has not been performed. We examined the frequency of complications identified by a complication screening method and their relationship to patient characteristics among congenital heart surgery admissions. Data were obtained from the Healthcare Cost and Utilization Project Kids' Inpatient Database 2000. Among congenital heart surgery admissions, age < or =20 years, we identified International Classification of Disease, 9th Revision, Clinical Modification (ICD-9-CM) codes indicating complication. Complication diagnoses were categorized as related to drug/biologic, procedures, devices, implants and grafts, and radiation. We used the Risk Adjustment for Congenital Heart Surgery risk categories (1-6) to examine the association between case complexity and complications. Multivariate analyses estimated the odds for a complication diagnosis by patient characteristics, including age, prematurity, chromosomal anomalies, noncardiac structural anomalies, and surgical risk category. Among 12,717 cases, 4014 (32%) had at least 1 complication diagnosis code. Procedure-related complication diagnoses represented 75% of complication diagnoses; device, implant, or graft represented 21%; drug/biologic represented 4% and radiation represented 0%. Multivariate analyses demonstrated that higher surgical case complexity and older age had a greater risk for a complication diagnosis: Risk Category 2, odds ratio (OR) 1.8; 3 OR 2.9; 4 OR 2.9; 5 OR 5.0; 6 OR 4.1, relative to category 1, all p < 0.01; age > or =12 years, OR 1.3, p < 0.001; <1 year OR 1.1, p = 0.31. Premature cases had decreased odds OR 0.4, p < 0.001. This complication screening method indicates that unwanted patient events occur frequently during congenital heart surgery admissions. Children undergoing complex congenital heart surgery are at greatest risk for these unwanted events. Further study of these events is needed to determine their preventability and severity. PMID- 17912482 TI - Measuring stress velocity index using mean blood pressure: simple yet accurate? AB - The stress velocity index, or the relationship of the rate-corrected mean velocity of circumferential shortening (VCFc) to the end systolic wall stress (ESWS), is a sensitive, load-independent measure of left ventricular contractility. ESWS is technically difficult to obtain and requires simultaneous blood pressure measurement, carotid artery tracing, and phonocardiogram. We report our comparison of two simpler methods of measuring ESWS and, therefore, stress velocity index. Patients with normal cardiac anatomy who had completed anthracycline chemotherapy were evaluated. ESWS as measured by the standard method using a carotid artery tracing (ESWScar) was compared to ESWS obtained using mean arterial pressure (ESWSmap) or systolic blood pressure (ESWSsbp). The cohort included 63 patients, with 37 (59%) males and a median age of 13.1 years. The mean (+/-SD) ESWScar was 53.3+/-15.3 g/cm(2) (range, 26.3-94 g/cm(2)); ESWSmap, 53 +/-13.4 g/cm(2) (range, 27.1-86.1 g/cm(2)); and ESWSsbp, 72.9 +/- 18.2 g/cm(2) (range, 40.8-117.2 g/cm(2)). ESWSmap and ESWSsbp closely correlated with ESWScar (coefficient correlation r = 0.88 and r = 0.87, respectively). Using ESWSmap, all patients were correctly classified as having normal or abnormal contractility as defined by stress velocity index, whereas ESWSsbp detected only two of the six patients with impaired contractility. We conclude that ESWSmap is a simple, highly sensitive and specific method for assessing left ventricular contractility. ESWSmap correlates closely with ESWScar and can be incorporated into the monitoring of cardiac dysfunction in the anthracycline-treated population. Further studies are needed to determine if this simplified measure accurately assesses the ESWS in other cardiac disease states. PMID- 17912483 TI - Spontaneous development and rupture of pulmonary artery aneurysm: a rare complication in an infant with peripheral pulmonary artery stenoses due to mutation of the elastin gene. AB - We present a 3-year-old boy with an elastin gene mutation and multiple peripheral pulmonary stenoses, who developed aneurysms of the pulmonary arteries spontaneously. We performed transcatheter occlusion of the aneurysms with detachable coils. While pulmonary arterial aneurysms may develop following pulmonary balloon angioplasty, spontaneous development is exceedingly rare. To the best of our knowledge, this is the first report describing spontaneous development of pulmonary artery aneurysms in a patient with peripheral pulmonary artery stenoses due to mutation of the elastin gene or Williams-Beuren syndrome. PMID- 17912484 TI - Fetal diagnosis of an "extra cardiac chamber". AB - Congenital right ventricular aneurysms and diverticula are rare congenital anomalies and prenatal detection is uncommon. Distinguishing between these two anomalies by imaging alone is difficult. Recently, we were asked to review a 21 week ultrasound that detected an "extra cardiac chamber." This represents the first report of an isolated out-pouching of the right ventricular free wall to be detected by prenatal screening ultrasound. Based on preoperative imaging, this out-pouching was diagnosed as a ventricular aneurysm, but intraoperative findings suggested it was a diverticulum. This case will review the differences between ventricular aneurysms and diverticula and illustrate the use and limitations of various imaging modalities used in their evaluation, both prenatally and postnatally. A plan of management is suggested. PMID- 17912485 TI - A unique case of reversible myocardial ischemia in a hyperthyroid neonate. AB - A case of an 8-day-old preterm neonate with heart failure, pulmonary hypertension, and myocardial ischemia due to hyperthyroidism is reported. Treatment of the disease initially with b-blockers and, upon establishment of hyperthyroidism, with propylthiouracil reversed all cardiac abnormalities. Contrary to the rule, diagnosis of hyperthyroidism in the mother was established following the diagnosis of the condition in her baby. On long-term follow-up (10 years), the child has developed normally, remains euthyroid with normal electrocardiogram. To our knowledge, diagnosis and reversal of the above mentioned ischemic abnormalities have not been previously reported in neonates. PMID- 17912486 TI - Photodynamic therapy following intravitreal bevacizumab in multifocal choroiditis. AB - PURPOSE: To report the use of intravitreal bevacizumab (avastin) followed by photodynamic therapy (PDT) in a case of choroidal neovascular membrane (CNVM) due to multifocal choroiditis. DESIGN: Interventional case report. METHODS: In the setting of institutional practice, a 40-year male patient with previously diagnosed multifocal choroiditis presented with reduced vision in his left eye. A CNVM was revealed through hazy vitreous in the left eye. Due to a prior episode of increased intraocular pressure with use of periocular steroid (triamcinolone), and marked vitreous haze precluding precise assessment of the neovascular membrane, intravitreal injection of bevacizumab was tried followed by PDT. RESULTS: There was clearing of the vitreous haze and reduction of leakage one week after intravitreal injection. PDT one month after injection resulted in stable vision at three-month follow-up without angiographic leakage. CONCLUSION: Intravitreal bevacizumab may be tried in cases of CNVM due to multifocal choroiditis. Timing of the injection may vary with the clinical scenario. PMID- 17912487 TI - Enhancement of the solubility and efficacy of poorly water-soluble drugs by hydrophobically-modified polysaccharide derivatives. AB - PURPOSE: This work was intended to develop and evaluate a new polymeric system based on amphiphilic carboxymethylpullulans (CMP(49)C(8) and CMP(12)C(8)) that can spontaneously self-assemble in aqueous solutions and efficiently solubilize hydrophobic drugs. METHODS: The self-assembling properties of CMP(49)C(8) and CMP(12)C(8) were characterized by fluorescence spectroscopy and surface tension measurements. The solubilization of benzophenone and docetaxel was assessed from surface tension measurements, UV spectrometry and HPLC assays. The in vitro cytoxicity of CMP(49)C(8) solutions and the docetaxel commercial vehicle (Tween 80/Ethanol-water) were evaluated in the absence and in the presence of docetaxel. RESULTS: Compared to CMP(12)C(8), CMP(49)C(8) in aqueous solutions appeared to self-organize into monomolecular aggregates containing hydrophobic nanodomains, and to significantly increase the apparent solubility of benzophenone. Docetaxel solubility could also be improved in the presence of CMP(49)C(8) but to a lower extent due to the surface properties of the drug. Nevertheless, in vitro, the cytotoxicity studies revealed that against cancer cells, the CMP(49)C(8) docetaxel formulation was equipotent to the commercial docetaxel one. Furthermore, in the absence of the drug, CMP(49)C(8) appeared less cytotoxic against macrophages than the Tween 80/Ethanol-water. CONCLUSIONS: CMP(49)C(8) is a good candidate for solubilizing hydrophobic drugs and could be applied to docetaxel formulations. PMID- 17912488 TI - Paclitaxel prodrugs with sustained release and high solubility in poly(ethylene glycol)-b-poly(epsilon-caprolactone) micelle nanocarriers: pharmacokinetic disposition, tolerability, and cytotoxicity. AB - PURPOSE: Develop a Cremophor and solvent free formulation of paclitaxel using amphiphilic block co-polymer micelles of poly(ethylene glycol)-b-poly(epsilon caprolactone) (PEG-b-PCL) and characterize their release, solubility, cytotoxicity, tolerability, and disposition. METHODS: Hydrophobic prodrugs of paclitaxel were synthesized via DCC/DMAP or anhydride chemistry to overcome the poor loading (<1% w/w) of paclitaxel in micelles of PEG-b-PCL. Micelles were prepared by a co-solvent extraction technique. A micellar formulation of paclitaxel prodrug (PAX7'C(6)) was dosed intravenously to rats (10 mg/kg) and compared to Taxol (paclitaxel in CrEL:EtOH) and PAX7'C(6) in CrEL:EtOH as controls at the same dose. Pharmacokinetic parameters and tissue distribution were assessed. RESULTS: Paclitaxel prodrugs had solubilities >5 mg/ml in PEG-b PCL micelles. Resulting PEG-b-PCL micelles contained 17-22% w/w prodrug and were less than 50 nm in diameter. PEG-b-PCL micelles released paclitaxel prodrugs over several days, t(1/2)>3 d. Only the 7'derivative of paclitaxel with the shortest acylchain 7'hexonoate (PAX7'C(6)) maintained cytotoxic activity similar to unmodified paclitaxel. PAX7'C(6) micelles demonstrated an increase in area under the curve, half-life, and mean residence time while total clearance and volume of distribution decreased. CONCLUSIONS: Paclitaxel prodrugs in PEG-b-PCL micelle nanocarriers augment the disposition and increase tolerability making further studies on tumor efficacy warranted. PMID- 17912489 TI - Delivery by cationic gelatin nanoparticles strongly increases the immunostimulatory effects of CpG oligonucleotides. AB - PURPOSE: Cationized gelatin nanoparticles (GNPs) were used as carrier to improve delivery of immunostimulatory CpG oligonucleotides (CpG ODN) both in vitro and in vivo. METHODS: Uptake of CpG ODN-loaded cationized gelatin nanoparticles (CpG GNPs) into murine myeloid dendritic cells (DCs) and their respective immunostimulatory activity was monitored. In vivo, induction of cytokine secretion by CpG-GNPs was measured. For experiments on primary human cells, prototypes of the three CpG ODN classes were adsorbed onto GNPs. Uptake and induction of proinflammatory cytokines were assessed in human plasmacytoid DCs and B cells, the only existing human target cells for CpG ODN. RESULTS: In the murine system, gelatin nanoparticle formulations enhanced the uptake and immunostimulatory activity of CpG ODN both in vitro and in vivo. Furthermore, delivery by cationized gelatin nanoparticles of CpG ODN of the classes B and C to primary human plasmacytoid DCs increased production of IFN-alpha, a key cytokine in the driving of both the innate and adaptive immune responses. CONCLUSION: GNPs can be used as a biodegradable and well tolerated carrier to deliver CpG ODN to their target cells and strongly increase activation of the immune system. This concept may be applied as novel adjuvant for antiviral and antitumoral vaccines. PMID- 17912490 TI - Increased serum levels of soluble CD30 in patients with common variable immunodeficiency and its clinical implications. AB - Common variable immunodeficiency (CVID) is a heterogeneous group of disorders, characterized by hypogammaglobulinemia and increased susceptibility to recurrent pyogenic infections, autoimmunity, and malignancies. Twenty-five cases with CVID (18 male and 7 female) and 25 healthy volunteers were investigate in this study. Soluble CD30 (sCD30) serum levels of the subjects were measured and compared. Serum levels of sCD30 in the patients with CVID were significantly increased in comparison with controls (36.93 +/- 32.38 vs 5.27 +/- 1.32 U/ml, P < 0.001). The group of patients with splenomegaly and reversed ratio of CD3+CD4+ T cells/CD3+CD8+ T cells had the highest serum levels of sCD30 (66.01 +/- 43.34 U/ml) in comparison with other patients (P = 0.010). High levels of sCD30 in the CVID patients with splenomegaly and the presence of lymphoma in a patient with the highest level of sCD30 may suggest a soluble form of this marker as a prognostic tool in such diseases. PMID- 17912491 TI - Intracerebral haemorrhage and recombinant factor VIIa: not so good news! PMID- 17912492 TI - Acute oppressive chest pain in a 66-year-old woman with a recent Amplatzer device closure of patent foramen ovale. PMID- 17912494 TI - Aggressions and size-related fecundity of queenless workers in the ant Cataglyphis cursor. AB - In social hymenoptera, the reproductive division of labor is often linked to differences in individual body size with the reproductive caste (the queen) being larger than the workers. Likewise, the reproductive potential may vary with size within the worker caste and could affect the evolution of worker size in social insects. Here, we tested the relationship between worker size and reproductive potential in the facultative parthenogenetic ant Cataglyphis cursor. Colonies are headed by a multiply mated queen, but workers can produce gynes (virgin queens) and workers by thelytokous parthenogenesis after the queen's death. We observed the behaviour of workers (n = 357) until the production of gynes (212 h over 3 months) in an orphaned colony (mated queen not present). The size of workers was measured, and their paternal lineage determined using six microsatellite markers, to control for an effect of patriline. Larger workers were more likely to reproduce and lay more eggs indicating that individual level selection could take place. However, paternal lineage had no effect on the reproductive potential and worker size. From the behavioural and genetic data, we also show for the first time in this species, evidence of aggressive interactions among workers and a potential for nepotism to occur in orphaned colonies, as the five gynes produced belonged to a single paternal lineage. PMID- 17912493 TI - Clockwork blue: on the evolution of non-image-forming retinal photoreceptors in marine and terrestrial vertebrates. AB - This paper presents a hypothesis that could explain why blue light appears to dominate non-image-forming (NIF) ocular photoreception in marine as well as terrestrial vertebrates. Indeed, there is more and more evidence suggesting that 'novel' retinal photoreceptors, which are sensitive to blue light and were only discovered in the 1990s, could be a feature shared by all vertebrates. In our view, blue light photoreception evolved and persisted as NIF photoreception because it has been useful in the colonisation of extensive photo-dependent oceanic habitats and facilitated the move of vertebrates from an aquatic to a terrestrial environment. Because the available scattered evidence is compatible with the validity of our hypothesis, we hope that our rationale will be followed up. Indeed, it (1) involves testable predictions, (2) provides plausible explanations for previous observations, (3) unites phenomena not previously considered related to one another and (4) suggests tests that have not been carried out before. Overall, our approach not only embraces cross-disciplinary links; it, moreover, serves as a reminder of an all-embracing evolutionary history, especially with regard to a ubiquitous photoreceptive 'clockwork-blue' in marine and terrestrial vertebrates. PMID- 17912496 TI - Identification of quantitative trait loci that affect endoreduplication in maize endosperm. AB - Endoreduplication in maize endosperm precedes the onset of starch and storage protein synthesis, and it is generally thought to influence grain filling. We created four backcross populations by reciprocally crossing the F(1) progeny of a cross between Sg18 and Mo17 to the parental inbreds, which differ in endoreduplication by two parameters--mean ploidy and percentage of endoreduplicated nuclei. This four-backcross design allowed us to estimate and test the additive and dominant genetic effects of quantitative trait loci (QTLs) affecting endoreduplication. An analysis of endosperm from the four backcross populations at 16 days after pollination using a modified triploid mapping approach identified three endosperm QTLs influencing mean ploidy and two endosperm QTLs affecting the percentage of endoreduplicated nuclei. Some of these QTLs may manifest their effects on endoreduplication via expression in the embryo. The QTLs detected display strong dominance or over-dominance and interacted epistatically with an embryo-expressed QTL. This helps to explain the genetic basis for transgressive segregation in the backcross progeny. Although the favorable alleles that increase mean ploidy and percentage of endoreduplicated nuclei can be contributed by both parents, the Mo17-derived alleles for endoreduplication were often dominant or over-dominant to the Sg18 derived allele. One QTL on chromosome 7 that may be expressed in both the embryo and endosperm exerted a pleiotropic effect on two different parameters of endoreduplication. The results from this study shed light on the regulation of endoreduplication in maize endosperm and provide a marker-assisted selection strategy for potentially improving grain yield. PMID- 17912497 TI - The effect of epinephrine by nebulization on measures of airway obstruction in patients with acute severe croup. AB - OBJECTIVES: To demonstrate that tests of pulmonary function applicable to sick infants and small children with acute severe viral croup would provide clear, objective evidence of responsiveness to therapy with nebulized epinephrine. STUDY DESIGN: Oesophageal pressure changes and airflows at the mouth were measured in 17 patients with acute severe croup, before and after nebulization with epinephrine. RESULTS: In 12 of the 17 patients there was a significant improvement in respiratory mechanics following epinephrine nebulization. Six of the 12 patients who responded to adrenaline also received 0.9% saline by nebulization, without improvement. No measures derived from combined flow and volume data showed any statistically significant change following epinephrine nebulization. Measures combining flow and pressure data, specifically inspiratory airway resistance, expiratory airway resistance, work of breathing, rate of work of breathing and volume for effort, showed changes of 26%, 33%, 16%, 16% and 46% respectively. The most statistically significant measures were pressure-rate product, pressure-time integral, oesophageal pressure alone and expiratory resistance. These changes persisted for at least 10 min after inhalation although there was some evidence of decline in pharmacologic effect at that time. CONCLUSIONS: Nebulized epinephrine results in a short-lived improvement in some but not all patients with croup. This reduction in respiratory effort occurs secondary to a decline in inspiratory and expiratory airway resistance. Oesophageal pressures measured via a feeding tube are satisfactory for quantification of the acute response and may be a useful continuous monitoring device. Flow measurements are unhelpful, and continuous administration of nebulized epinephrine should be investigated. PMID- 17912495 TI - [Urological problems in patients with meningomyelocele. Diagnostic studies and management]. AB - Since the 1980s the management of children and adolescents with meningomyelocele has undergone major changes. The introduction of pharmacotherapy with antimuscarinic agents, clean intermittent catheterization (CIC) and antibacterial prophylaxis has revolutionized the management of children with neurogenic bladder. The co-operation between neonatologists, neurosurgeons, paediatric neurologists, paediatricians, paediatric urologists, paediatric nephrologists, paediatric orthopaedists and paediatric surgeons is necessary to achieve an optimized therapy in each individual patient. In this interdisciplinary consensus paper we provide definitions and classifications as well as a timetable for the appropriate investigations. The conservative and surgical options are explained in detail. A short review is given concerning orthopaedic management, incidence of latex allergy, options for bowel management, diagnosis and treatment of urinary tract infections, problems with sexuality and fertility as well as the long-term compliance of these patients and their relatives. PMID- 17912498 TI - Sulfotransferase 1A1 and glutathione S-transferase P1 genetic polymorphisms modulate the levels of urinary 8-hydroxy-2'-deoxyguanosine in betel quid chewers. AB - Betel quid chewing has been associated with several human cancers. However, the role of betel quid in carcinogenesis remains uncertain. Piper betel contains high concentrations of safrole (an inducer of DNA oxidative damage). Safrole may be metabolized by hepatic sulfotransferase 1A1 (SULT1A1), or glutathione S transferases (GSTM1, GSTT1, and GSTP1). Thus, we investigated the association of genetic polymorphisms of SULT1A1, GSTM1, GSTT1, and GSTP1 with DNA oxidative damage among betel quid chewers. A biomarker for oxidative stress, urinary 8 hydroxy-2'-deoxyguanosine (8-OHdG) level, was analyzed using isotope-dilution LC MS/MS in 64 betel quid chewers and 129 non-betel quid chewers. Data on demographics and habits (smoking, alcohol drinking, and betel quid chewing) were obtained from questionnaires. Our results revealed that urinary 8-OHdG level was higher in chewers with SULT1A1 Arg-His genotype than in chewers with SULT1A1 Arg Arg genotype. Urinary 8-OHdG level was also higher in chewers with GSTP1 Ile-Ile genotype. Furthermore, the combined effect of SULT1A1 and GSTP1 genotypes on urinary 8-OHdG was evaluated. Non-chewers with both SULT1A1 Arg-Arg and GSTP1 Val Val/Ile-Val (reference group) had the lowest mean level (3.6 ng/mg creatinine), whereas chewers with either SULT1A1 Arg-His or GSTP1 Ile-Ile had the highest 8 OHdG mean level (6.2 ng/mg creatinine; vs. reference group, P = 0.04). Chewers with both of SULT1A1 Arg-Arg and GSTP1 Val-Val/Ile-Val (4.6 ng/mg creatinine), and non-chewers with either SULT1A1 Arg-His or GSTP1 Ile-Ile (4.7 ng/mg creatinine) had a moderately increased 8-OHdG level. Thus, the susceptible SULT1A1 and GSTP1 genotypes may modulate increased DNA oxidative stress elicited by betel-quid chewing. PMID- 17912499 TI - Galantamine and donepezil differently affect isolation rearing-induced deficits of prepulse inhibition in mice. AB - RATIONALE: Previous studies have shown that alterations in acetylcholine (ACh) receptor subtypes might contribute to cognitive impairment observed in schizophrenia and that choline acetyltransferase activity in the parietal cortex is negatively correlated with the severity of such cognitive impairment. However, clinical data suggest that the acetylcholinesterase (AChE) inhibitors galantamine and donepezil have different effects on negative and cognitive symptoms in schizophrenia. Prepulse inhibition (PPI) deficits--sensory information-processing deficits observed in schizophrenia--may be useful models for studying the efficacy of AChE inhibitors as cognitive enhancers. OBJECTIVES: The present study examined the effects of galantamine and donepezil on PPI deficits induced by an environmental factor and drugs. MATERIALS AND METHODS: In the isolation-rearing model, 3-week-old male ddY mice were housed either in groups of five or six per cage or isolated in cages of the same size for more than 6 weeks. In the drug induced model, apomorphine 1 mg/kg and MK-801 0.2 mg/kg were administered to 9- to 10-week-old male ddY mice. RESULTS: In isolation-reared mice, galantamine attenuated PPI deficits, while donepezil did not. Galantamine and donepezil both attenuated PPI deficits induced by apomorphine, but not by MK-801. The galantamine-induced improvements in PPI deficits were not prevented by the nicotinic ACh receptor antagonists mecamylamine and methyllycaconitine. CONCLUSIONS: These observations suggest that galantamine and donepezil have different effects on the environmentally induced PPI deficits and that these observations may be relevant to the different effects of these drugs observed clinically in schizophrenia. PMID- 17912502 TI - Chemically driven variable selection by focused multimodal genetic algorithms in mid-IR spectra. AB - Four genetic-algorithm-based approaches to variable selection in spectral data sets are presented. They range from a pure black-box approach to a chemically driven one. The latter uses a fitness function that takes into account not only typical parameters like the number of errors when classifying a training set but also the chemical interpretability of the selected variables. In order to cope with the fact that multiple solutions may be acceptable, a multimodal genetic algorithm (GA) is employed and the most satisfactory solution selected. The multimodal GA uses two populations (denominated "hybrid two populations" GA or HTP-GA): a classical population, from which potential solutions emerge, and a new population, which maintains diversity in the search space (as required by multimodal problems). Results show that the HTP-GA approach improves the chemical understanding of the selected solution (compared to other GA approaches) and that the classification capabilities of the approach are still good. All of the GA strategies for variable selection were compared with a classical parametric technique, Procrustes rotation, which does not consider interpretability. PMID- 17912501 TI - Dopamine D2 receptor occupancy levels of acute sulpiride challenges that produce working memory and learning impairments in healthy volunteers. AB - RATIONALE: In humans, the effects of dopaminergic agents administered systemically are less clear-cut than studies in experimental animals where agents can be applied locally in the brain. DA receptor occupancy could clearly contribute to the variance in findings, although this is typically not known. OBJECTIVES: The objective of the study was to measure the DA D2 receptor occupancy of sulpiride 200 and 400 mg and relate this to changes in task performance. MATERIALS AND METHODS: Positron emission tomography scans were acquired in ten healthy volunteers with [11C]-raclopride. Striatal drug occupancy was calculated as the percentage change in binding potential between placebo and drug scans. All volunteers received placebo and sulpiride 400 mg, with four receiving 200 mg on a third session. Immediate post-scan neuropsychological assessment included working memory and learning tasks. RESULTS: Striatal sulpiride occupancy was approximately 17% (200 mg) and approximately 28% (400 mg), with similar occupancy within the midbrain. Neuropsychological data analysis was restricted to the higher dose (n = 10). Accuracy on the spatial working memory and spatial learning tasks was impaired after the drug, and the former was inversely related to occupancy. CONCLUSION: Doses of sulpiride typically used in human cognitive studies produced low levels of DA D2 receptor occupancy compared to that considered efficacious in the treatment of schizophrenia. The levels of occupancy were sufficient to replicate impairments on a spatial working memory task and impair spatial learning. The relationship between occupancy and working memory was suggestive of presynaptic effects, although the precise mechanism underlying the impairment will require studies of wider ranges of occupancy within and outside of the striatum. PMID- 17912503 TI - Reduction of the interferences of biochemicals and hematocrit ratio on the determination of whole blood glucose using multiple screen-printed carbon electrode test strips. AB - A practical approach to reduce the interferences of biochemicals and hematocrit ratio (Hct%) in the determination of whole blood glucose using multiple screen printed carbon electrode (SPCE) test strips is described. SPCE test strips with and without glucose oxidase [i.e., GOD(+)-SPCEs and GOD(-)-SPCEs] were used and the chronoamperometric currents of test glucose solutions with various spiked uric acid concentrations and Hct% were measured. By establishing the interference relationships between glucose concentrations and uric acid concentrations as well as Hct% values and with appropriate corrections, the whole blood glucose determinations could be made to be more accurate and comparable to those determined by the reference YSI method. Specifically, the use of the DeltaI value, i.e., the current difference between GOD(+)-SPCE and GOD(-)-SPCE measurements, would reduce most of the uric acid/biochemical interferences. An interpolation method was also established to correct for the glucose determinations with Hct% interferences. The Hct% corrections using the interpolation method are especially important and necessary for those blood samples with glucose concentrations higher than 110 mg dL(-1) and Hct% values lower than 35%. This approach should also be applicable to other biochemical determinations using similar electrochemical techniques. PMID- 17912500 TI - Effect of transdermal nicotine replacement on alcohol responses and alcohol self administration. AB - RATIONALE: Nicotine replacement is commonly used to treat tobacco use in heavy drinking smokers. However, few studies have examined the effect of nicotine replacement on subjective and physiological responses to alcohol and alcohol drinking behavior. OBJECTIVE: The primary aim of this within-subject, double blind study was to examine whether transdermal nicotine replacement (0 mg vs 21 mg/day) altered response to a low-dose priming drink and subsequent ad libitum drinking behavior. MATERIALS AND METHODS: Subjects (n=19) were non-treatment seeking, non-dependent heavy drinkers who were daily smokers. Six hours after transdermal patch application, subjective and physiological responses to a priming drink [designed to raise blood alcohol levels (BALs) to 0.03 g/dl] were assessed. This was followed by a 2-h self-administration period where subjects could choose to consume up to eight additional drinks (each designed to raise BALs by 0.015 g/dl) or to receive monetary reinforcement for drinks not consumed. RESULTS: We found that 6 h after patch application, tobacco craving associated with withdrawal relief was decreased, and systolic blood pressure and heart rate were increased in the active patch condition compared to the placebo patch condition. Subjective intoxication in response to the priming drink was attenuated in the active nicotine patch condition compared to 6 h of nicotine deprivation (i.e., placebo patch). During the self-administration period, subjects had longer latencies to start drinking and consequently appeared to consume fewer drinks when administered the active patch compared to the placebo patch. CONCLUSIONS: In heavy drinkers, transdermal nicotine replacement compared to mild nicotine deprivation attenuated subjective and physiological alcohol responses and delayed the initiation of drinking. PMID- 17912505 TI - Constraints on visuo-motor adaptation depend on the type of visual feedback during practice. AB - Adaptation to a novel visuo-motor gain has been found to generalize across target directions, whereas simultaneous adaptation to different direction-related visuo motor gains turned out to be impossible. We ask whether this is a rigid constraint on human adaptability or a soft constraint that can be overcome by optimized conditions of practice. In particular, we compared practice with continuous visual feedback, as used in previous studies, to practice with terminal visual feedback. With terminal visual feedback only the final positions of the movements are shown. Both kinds of visual feedback in principle can serve the acquisition of an internal model of direction-related visuo-motor gains, but with continuous feedback on-line visual closed-loop control permits accurate movements without access to an internal model. Whereas we found no indication of visuo-motor adaptation after continuous-feedback practice, there was adaptation after terminal-feedback practice. This was revealed both by (direction-related) adaptive shifts of movement amplitudes in an open-loop test with cued visuo-motor transformation and by (direction-related) aftereffects in an open-loop test with absence of the visuo-motor transformation being cued. None of the two groups gave evidence of explicit knowledge of the direction-related visuo-motor gains. These findings show that constraints on human adaptability can depend on the kind of experience with visuo-motor transformations, in particular on the kind of feedback during practice. PMID- 17912504 TI - Vertical eye position responses to steady-state sinusoidal fore-aft head translation in monkeys. AB - A major function of the otolith organ is to detect linear acceleration generated by two different head conditions, dynamic linear translation and static tilt relative to gravity. To investigate these sensory functions of the otolith organ, we analyzed vertical eye position in response to steady-state sinusoidal fore-aft translation over a range of frequencies (0.5-4 Hz) and amplitudes (0.10-0.33 g) in three monkeys. Vertical vestibuloocular reflexes elicited by linear acceleration (LVORs) during sinusoidal fore-aft translation were divided into translational LVOR component and tilt LVOR component taking vertical gaze dependent properties into account. Based on geometrical considerations, the translational LVOR component, but not the tilt LVOR component, depended on vertical gaze eccentricity. To quantify these two components, we used a V-shaped function model, plotting vertical eye sensitivities (deg/cm) against vertical gaze eccentricities (deg). The slope (deg/cm per degree) and intercept (sensitivity at zero gaze eccentricity) of this function approximately reflected the translational and tilt LVOR components, respectively. Our data show that the tilt LVOR component is independent of the reciprocal of the fixation distance (MA), whereas the translational LVOR component is almost linearly related to MA. The gain of the tilt LVOR component, characterized by low-pass dynamics, was greatest (0.36) at 0.5 Hz. Visual information clearly reduced the gain of the tilt LVOR component, by approximately 50%. There was no difference between the effects of large-field and small-spot stimuli. These findings demonstrate that steady-state sinusoidal fore-aft translation at lower frequencies stimulates the otolith organs and produces a pseudo-pitch tilt in cooperation with the gravito inertial force and as a result elicits an ocular response equivalent to the tilt LVOR. PMID- 17912506 TI - The metabolism of serotonin in neuronal cells in culture and platelets. AB - The aim of this study is to find a relationship between serotonin (5-HT) and its metabolite 5-hydroxy indol acetic acid (5-HIAA) in hippocampus, frontal neocortex and platelets. Serotonin and 5-HIAA were measured in cultured neurons and compared with those produced by human platelets. The cortical neuronal 5 HIAA/serotonin ratio was 4.7 and for hippocampal neurons it was 3.2. In human platelets, this ratio was 1.35 suggesting that the highest serotonin metabolism occurs in the frontal neocortex followed by the hippocampus and platelets. In the presence of 0.3 microM of p-chlorophenylalanine both cultured neurons and platelets exhibited an approximately 50% decrease in serotonin and 5-HIAA concentration suggesting similarities in the metabolic profile in both preparations. In addition, we found that serotonin by itself does not play any role in platelet aggregation but potentiates this phenomenon in the presence of calcium ionophore A23187. This synergistic interaction between serotonin (2-5 microM) and A23187 (0.5-2 microM) was inhibited by serotonin receptor blockers [methysergide (IC50 = 18 microM) and cyproheptadine (IC50, 20 microM)] and calcium channel blockers (verapamil and diltiazem, IC50 = 20 and 40 microM, respectively) that indicate both mechanisms are receptor mediated. Similarly, U73122, an inhibitor of phospholipase C (PLC), blocked the synergistic effect of serotonin and ionophore at an IC50 value of 9.2 microM. Wortmannin, a phosphoinositide 3-kinase (PI 3-K) inhibitor, also blocked the response (IC50 = 2.6 microM) by inhibiting respiratory burst. However, neither genistein, a tyrosine-specific protein kinase inhibitor, nor chelerythrine, a protein kinase C (PKC) inhibitor, affected aggregation. Our results are strongly suggestive of a synergistic interaction between serotonin type-2 and Ca-ionophore via a PLC/Ca signalling pathway. PMID- 17912507 TI - Anticipatory postural adjustments in arm muscles associated with movements of the contralateral limb and their possible role in interlimb coordination. AB - While sitting on a turnable stool, with both shoulders flexed at 90 degrees or, alternatively, with arms parallel to the trunk and the elbows flexed at 90 degrees--the hands being semisupine--subjects performed unidirectional and cyclic movements on the horizontal plane of the right arm (adduction-abduction) or hand (flexion-extension). The left arm was still, in a position symmetrical to that of the right limb and with the hand contacting a fixed support by the palmar or dorsal surface. During both unidirectional and cyclic arm or hand movements, activation of the prime mover muscles (right Pectoralis Major for arm adduction and Infraspinatus for abduction; right Flexor Carpi Radialis and Extensor Carpi Radialis for the hand movements) was accompanied by activation of the homologous muscles of the contralateral arm and inhibition of antagonists. The contralateral activities (1) regularly preceded the burst in the movement prime movers and (2) were organised in fixation chains that, exerting forces on the hand fixed support, will counterbalance the rotatory action exerted on the trunk by the primary movement. Based on these features, these activities may be classified as anticipatory postural adjustments (APAs). The observed APAs distribution is such as to favour the preferential (mirror symmetrical) coupling of upper limb movements on the horizontal plane. The possible role of these APAs in determining the different constraints experienced when performing mirror symmetrical versus isodirectional coupling is discussed. PMID- 17912509 TI - Yersinia enterocolitica-associated generalized microinfarctions of bone and spleen in a child. AB - We report a case of unusual extraintestinal yersiniosis in a 16-year-old girl with generalized microinfarctions of the bone and spleen. For the past 2 years she had been repeatedly admitted to our hospital with reactive arthritis, erythema nodosum and iridocyclitis of unknown aetiology. Ultrasound showed multiple round hypoechoic lesions in the spleen that were shown to have low T2 signal on MRI. MRI also showed disseminated nodular lesions of the skeleton that were low T1 and high T2 signal and demonstrated inhomogeneous contrast enhancement. The patient is currently in good health on low-dose nonsteroidal immunosuppressive therapy. This is a unique case of microinfarctions of the skeleton and spleen caused by a severe postinfectious autoimmune reaction following extraintestinal Yersinia enterocolitica infection. PMID- 17912508 TI - Role of lateral muscles and body orientation in feedforward postural control. AB - The study investigates the role of lateral muscles and body orientation in anticipatory postural adjustments (APAs). Subjects stood in front of an aluminum pendulum and were required to stop it with their right or left hand. An experimenter released the pendulum inducing similar body perturbations in all experimental series. The perturbation directions were manipulated by having the subjects standing on the force platform with different body orientations in relation to the pendulum movements. Consequently, perturbations were induced in sagittal, oblique, and frontal planes. Ground reaction forces and bilateral EMG activity of dorsal, ventral, and lateral trunk and leg muscles were recorded and quantified within the time intervals typical of APAs. Anticipatory postural adjustments were seen in all experimental conditions; their magnitudes depended on the body orientation in relation to the direction of perturbation. When the perturbation was produced in the lateral and oblique planes, APAs in the gluteus medius muscles were greater on the side opposite to the side of perturbation. Conversely, simultaneous anticipatory activation of the external obliques, rectus abdominis, and erector spinae muscles was observed on the side of perturbation when it was induced in the lateral plane. The results of the present study provide additional information on the directional specificity of anticipatory activation of ventral and dorsal muscles. The findings provide new data on the role of lateral muscles in feedforward postural control and stress the importance of taking into consideration their role in the control of upright posture. PMID- 17912510 TI - Engineering the phenylacetaldehyde reductase mutant for improved substrate conversion in the presence of concentrated 2-propanol. AB - Phenylacetaldehyde reductase (PAR) from Rhodococcus sp. ST-10 is useful for chiral alcohol production because of its broad substrate specificity and high stereoselectivity. The conversion of ketones into alcohols by PAR requires the coenzyme NADH. PAR can regenerate NADH by oxidizing additional alcohols, especially 2-propanol. However, substrate conversion by wild-type PAR is suppressed in concentrated 2-propanol. Previously, we developed the Sar268 mutant of PAR, which can convert several substrates in the presence of concentrated 2 propanol. In this paper, further mutational engineering of Sar268 was performed to achieve higher process yield. Each of nine amino acid positions that had been examined for generating Sar268 was subjected to saturation mutagenesis. Two novel substitutions at the 42nd amino acid position increased m-chlorophenacyl chloride (m-CPC) conversion. Moreover, several nucleotide substitutions identified from libraries of random mutations around the start codon also improved the PAR activity. E. coli cells harboring plasmid pHAR1, which has the integrated sequence of the top clones from the above selections, provided greater conversion of m-CPC and ethyl 4-chloro-3-oxobutanoate than the Sar268 mutant, with very high optical purity of products. This mutant is a promising novel biocatalyst for efficient chiral alcohol production. PMID- 17912511 TI - An update on microbial carotenoid production: application of recent metabolic engineering tools. AB - Carotenoids are ubiquitous pigments synthesized by plants, fungi, algae, and bacteria. Industrially, carotenoids are used in pharmaceuticals, neutraceuticals, and animal feed additives, as well as colorants in cosmetics and foods. Scientific interest in dietary carotenoids has increased in recent years because of their beneficial effects on human health, such as lowering the risk of cancer and enhancement of immune system function, which are attributed to their antioxidant potential. The availability of carotenoid genes from carotenogenic microbes has made possible the synthesis of carotenoids in non-carotenogenic microbes. The increasing interest in microbial sources of carotenoid is related to consumer preferences for natural additives and the potential cost effectiveness of creating carotenoids via microbial biotechnology. In this review, we will describe the recent progress made in metabolic engineering of non carotenogenic microorganisms with particular focus on the potential of Escherichia coli for improved carotenoid productivity. PMID- 17912512 TI - MRI appearance of surgically proven abnormal accessory anterior-inferior tibiofibular ligament (Bassett's ligament). AB - OBJECTIVE: A thickened accessory anterior-inferior tibiofibular ligament (Bassett's ligament) of the ankle can be a cause of ankle impingement. Its imaging appearance is not well described. The purpose of this study was to determine if the ligament could be identified on magnetic resonance imaging (MRI), to determine associated abnormalities, and to determine if MRI could be used to differentiate normal from abnormal. MATERIALS AND METHODS: Eighteen patients with a preoperative ankle MRI and an abnormal Bassett's ligament reported at surgery were found retrospectively. A separate cohort of 18 patients was selected as a control population. The presence of Bassett's ligament and its thickness were noted. The integrity and appearance of the lateral ankle ligaments, talar dome cartilage, and anterolateral gutter were also noted. RESULTS: In 34 of the 36 cases (94%), Bassett's ligament was identified on MRI. The ligament was seen in all three imaging planes and most frequently in the axial plane. The mean thickness of the ligament in the surgically abnormal cases was 2.37 mm, compared with 1.87 mm in the control with a p value=0.015 (t test). Nine of the 18 abnormal cases (50%) had talar dome cartilage lesions as a result of contact with the ligament at surgery, with only 3 cases of high-grade defects seen on MRI. Fourteen of the 18 abnormal cases (78%) had of synovitis or scarring in the lateral gutter at surgery, with only 5 cases with scarring seen on MRI. The anterior-inferior tibiofibular ligament was abnormal or torn in 8 of the 18 abnormal cases (44%) by MRI and confirmed in only 3 cases at surgery. DISCUSSION: Bassett's ligament can be routinely identified on MRI and was significantly thicker in patients who had it resected at surgery. An abnormal Bassett's ligament is often present in the setting of a normal anterior-inferior tibiofibular ligament. The cartilage abnormalities and synovitis associated with an abnormal Bassett's ligament are poorly detected by conventional MRI. PMID- 17912513 TI - Spectrum of shoulder injuries in the baseball pitcher. AB - This review describes a range of shoulder injuries experienced by baseball pitchers. It is estimated that more than 57% of pitchers suffer some form of shoulder injury during a playing season. Knowledge of the overhead throwing cycle is crucial for our understanding of these shoulder injuries. Baseball pitchers are prone to rotator cuff tears from tensile overload and impingement. Glenoid labrum degeneration or tears are also common, due to overuse syndrome (micro instability), internal impingement and microtrauma. An understanding of the lesions involved in overhead throwing is crucial in baseball pitchers, as long term disability can result from these injuries, sometimes with severe financial consequences to the player. PMID- 17912514 TI - A prospective analysis of CT density measurements of bone metastases after treatment with zoledronic acid. AB - OBJECTIVE: The objective was to prospectively determine CT density changes in bone metastases, before and after intravenous zoledronic acid for a maximum period of 12 months. PATIENTS AND METHODS: Twenty-three consecutive patients presented with bone metastases and underwent therapy with zoledronic acid from December 2004. All patients underwent CT of the chest, abdomen, and pelvis. Bone density, measured in Hounsfield units (HU), was determined by segmenting lesions in the same anatomical area of the metastasis sites on the axial images of the sequential series of CT examinations. The effects of zoledronic acid were evaluated by calculating absolute and relative increases in bone density. RESULTS: The patients presented with multiple metastases in 65% of the cases. When compared with the baseline, all groups demonstrated a significant increase in bone density, which significantly (p < 0.01) correlated with the number of zoledronic acid administrations. There was increased bone density of at least 100% in 57%, and an increase of at least 50% in 87% of the patients. This increase was significant in both lytic and sclerotic metastases after 3 months of therapy. No significant bone density difference was found in normal-appearing bone. CONCLUSION: Bone density measured by CT increases at metastatic sites after zoledronic acid treatment, regardless of the type of metastasis, in contrast to apparently normal bone. PMID- 17912515 TI - Are radiographs needed when MR imaging is performed for non-acute knee symptoms in patients younger than 45 years of age? AB - OBJECTIVE: The objective was to determine the value of radiographs in young adults with non-acute knee symptoms who are scheduled for magnetic resonance imaging (MRI). MATERIALS AND METHODS: Nine hundred and sixty-one consecutive patients aged between 16 and 45 years with knee symptoms of at least 4 weeks' duration were prospectively included in three participating hospitals. After applying exclusion criteria, 798 patients remained. Exclusion criteria were previous knee surgery (including arthroscopy) or MRI, history of rheumatoid arthritis, clinical diagnosis of retropatellar chondromalacia, contra-indication for MRI and recent trauma. We identified two groups: group A with no history of trauma (n = 332), and group B with an old (>4 weeks) history of trauma (n = 466). Patients had a standardized history taken, and underwent a physical exam, antero posterior (AP) and lateral radiographs and MRI. We evaluated the radiographs and MRI for osseous lesions, articular surface lesions, fractures, osteoarthritis, loose bodies, bone marrow edema and incidental findings. Subsequently, patients with osseous abnormalities (Kellgren grade 1 and 2 excluded) on radiographs and a matched control group was evaluated again using MRI without radiographs. RESULTS: Median duration of symptoms was 20 weeks. In group A, radiographs showed 36 osseous abnormalities in 332 patients (10.8%). Only 13 of these, all Kellgren grade 1 osteoarthritis, were not confirmed on MRI. MRI showed 72 (21.7%) additional abnormalities not confirmed on radiographs. In group B, radiographs showed 40 osseous abnormalities (8.6%) in 466 patients. Only 15 of these, all Kellgren grade 1 osteoarthritis, were not confirmed on MRI. MRI showed 194 (41.6%) additional abnormalities not confirmed on radiographs. The second evaluation of MRI without radiographs in 34 patients was identical to the first MRI evaluation. Common lesions were significantly more often diagnosed with MRI than with radiographs. CONCLUSION: Radiographs should not be obtained routinely when MRI is being performed in young adults with non-acute knee complaints because the yield and added value to MRI are low. PMID- 17912516 TI - Disc space-targeted angled axial MR images of the lumbar spine: a potential source of diagnostic error. AB - OBJECTIVE: To compare the sensitivities of contiguous axial (CA) images and disc space-targeted angled axial (DSTAA) images of the lumbar spine for: (1) the detection of spondylolysis defects, and (2) the identification of disc material which has migrated away from the parent disc. MATERIALS AND METHODS: Prospective imaging of the lumbar spine was performed over a 22-month period in 103 patients. Imaging protocols included spin-echo T1- and fast spin-echo (FSE) T2-weighted images in the sagittal and axial planes. For each patient, axial images were obtained both contiguously throughout the lumbar spine and as angled images, targeted at the region of the disc space only. Two separate data sets were compiled: one that included contiguous axial images (CA data set) and another that included targeted angled images through the disc spaces only (DSTAA data set). Identical sagittal images were included with both sets. A single radiologist independently interpreted the two data sets for a given patient following an intervening time lapse. The radiologist was blinded to the initial interpretation. Results from the two independent interpretations were then compared. RESULTS: Spondylolysis defects were identified at 15 different levels in 14 patients (14%) using the contiguous axial imaging protocol compared with 8 (7%) identified using the disc space targeted angled axial imaging protocol (P = 0.12). Migrated disc material was identified in 12 patients (12%) using the CA protocol compared with 3 patients (3%) identified with the DSTAA protocol (P = 0.016). CONCLUSIONS: MR imaging of the lumbar spine using contiguous axial data obtained through both the level of the disc and vertebral body demonstrates migrated disc material and spondylolysis defects better than did disc space targeted angled data. PMID- 17912517 TI - Soft tissue mass of the thigh: presentation, diagnosis and discussion. PMID- 17912518 TI - Temporal progression of skeletal cystic angiomatosis. AB - Cystic angiomatosis is a rare, benign, multifocal disorder of bone and viscera, in which angiomatous deposits of both vascular and lymphatic elements result in bone lysis and organ dysfunction. We report on a case of late-onset cystic angiomatosis in a Caucasian woman who first presented at age 35 years with a lytic expansile lesion of the proximal humerus, initially diagnosed as low-grade hemangio-endothelioma. This was treated with injection of cement and prophylactic pinning. However, the lesion continued to grow, and, 5 years later, she was discovered to have disseminated bony involvement, initially thought to represent metastatic disease. However, further investigation revealed a diagnosis of cystic angiomatosis, and the patient was treated with bisphosphonates. Follow-up over a 15-year period since her initial presentation at age 35 years has shown osteosclerotic conversion of many of the lesions, with development of numerous pathologic stress fractures that have failed to heal, despite operative intervention. PMID- 17912519 TI - Bone graft substitutes. PMID- 17912520 TI - Irreducible anterior dislocation of the shoulder due to soft tissue interposition of subscapularis tendon. AB - The shoulder joint is the most unstable joint in the body and is easily dislocated. Anterior shoulder dislocation is the commonest and can be associated with glenoid and humeral fractures. Anterior shoulder dislocations are not infrequently associated with cuff tendon tears; however, anterior shoulder dislocation is easily reducible. Irreducible anterior dislocation of the glenohumeral joint is uncommon, and this could be due to bony as well as soft tissue causes. Persistent anterior dislocation due to torn subscapularis interposition in the glenohumeral joint is very rare, and only a few operative cases have been reported in the literature. We present MR features of one such case and a literature review. PMID- 17912521 TI - Elimination of hand-wrist radiographs for maturity assessment in children needing orthodontic therapy. AB - Our aim was to evaluate the validity of the cervical vertebra maturation (CVM) method as an indicator of skeletal age during the circumpubertal period by correlating the CVM method with the hand-wrist maturation (HWM) method in an attempt to eliminate the need for hand-wrist radiographs for maturity assessment. Hand-wrist and lateral cephalometric radiographs of 400 Chinese were randomly selected. The age for girls was between 10 years and 15 years and for boys it was between 12 years and 17 years, so that they were within the circumpubertal period. The CVM was assessed by a method developed by Baccetti and co-workers, whereas hand-wrist maturation was assessed by Fishman's method. The CVM was significantly correlated with HWM skeletal age. (Spearman's r boys = 0.9206, girls = 0.9363). All the patients in cervical vertebra stage 3 (CVS3) of CVM corresponded to skeletal maturation indicator 2 (SMI2) or SMI3 stages of HWM (around the peak of the growth spurt). The method error was insignificant. CVM is a valid indicator of skeletal growth during the circumpubertal period. This work will provide dental practitioners with information on jaw growth modification therapy. PMID- 17912522 TI - Three-dimensional spin-lock magnetic resonance imaging of the shoulder joint at 3 T: initial experience. AB - OBJECTIVES: This was a pilot study which aimed to assess the feasibility of 3D spin-lock (3D-T(1rho)) MRI of the shoulder joint and to establish baseline values of healthy humeral and glenoid cartilages in vivo. MATERIAL AND METHODS: Four asymptomatic volunteers [mean age 31 years (range 29-36 years)] were recruited. A 3.0 T scanner, employing a four-channel, phased-array, shoulder, radio-frequency (RF) coil was used. Three-dimensional T(1rho)-weighted images were acquired with a 3D gradient-echo (GRE) sequence with T(1rho) magnetization preparation. In order to a construct T(1rho) map, we acquired four 3D-T(1rho)-weighted images with spin-locking length (TSL) values of 2 ms, 10 ms, 20 ms, and 30 ms. The glenoid and humeral cartilage were segmented manually at each slice of the 3D images. We performed additional regional analysis by dividing the cartilage into anterior/posterior and superior/inferior regions. RESULTS: The global average T(1rho) value of the shoulder cartilages varied from 37.9 ms to 48.5 ms and from 32.4 ms to 36.9 ms for humeral and glenoid cartilages, respectively. In the humeral cartilage, the average regional T(1rho) values varied from 35.9 ms to 52.2 ms; 54.4 ms to 69.0 ms; 39.1 ms to 49.3 ms and 34.6 ms to 57.2 ms for the anterior-superior, anterior-inferior , posterior-superior and posterior-inferior regions, respectively. In the glenoid cartilage, the values varied from 31.3 ms to 40.8 ms; 34.1 ms to 35.3 ms; 26.7 ms to 37.2 ms and 32.8 ms to 35.7 ms for the same regions, respectively. CONCLUSION: We demonstrated that 3D-T(1rho) MRI of the shoulder can be performed on a 3 T clinical scanner within specific absorption rate (SAR) limits, and we present baseline values for healthy patients which may be useful for quantitative comparison with diseased shoulders. PMID- 17912523 TI - Determinants of the response of left ventricular ejection fraction to vasodilator stress in electrocardiographically gated (82)rubidium myocardial perfusion PET. AB - PURPOSE: Myocardial perfusion imaging with (82)Rb PET allows for ECG-gated studies to be obtained early after radiotracer injection, capturing ventricular function close to peak pharmacologic action of dipyridamole. This is different from gated SPECT and may potentially provide additional diagnostic information. We sought to identify potential correlates of the PET-derived ejection fraction response to vasodilator stress. METHODS: One hundred ten consecutive patients undergoing (82)Rb PET myocardial perfusion imaging during evaluation for coronary artery disease were included. Using a GE Discovery STRx PET-CT scanner, ECG-gated images (eight bins) were obtained at rest and 4 min after dipyridamole infusion, 90 s after infusion of 1,480-2,220 MBq of (82)Rb. Summed rest, stress, and difference scores (SRS, SSS, and SDS) were determined using a five-point scoring system and 20-segment model. Ejection fraction was calculated using automated QGS software. RESULTS: Significant reversibility (SDS > or = 4) was found in 23 patients (21%). Mean LVEF in all patients was 47 +/- 13% at rest and 53 +/- 13% during dipyridamole. LVEF increased in 89 patients, and decreased in 17 patients during vasodilation. The change in LVEF was inversely correlated with SDS (r = 0.26; p = 0.007). Additionally, it was inversely correlated with resting LVEF (r = -0.20; p = 0.03) and SSS (r = -0.25; p = 0.009). No significant correlations were observed with SRS, heart rate, blood pressure, age, hypertension, hypercholesterolemia, or pretest likelihood of disease. At multivariate regression analysis, SDS was an independent predictor of the change in LVEF. CONCLUSIONS: Gated (82)Rb PET during pharmacologic stress allows for assessment of the functional response to vasodilation. The magnitude of LVEF increase is determined by stress perfusion/reversible perfusion defects. Functional response to hyperemia may thus be incorporated in future evaluations of diagnostic and prognostic algorithms based on (82)Rb PET. PMID- 17912524 TI - PET/CT with Gluc-Lys-([(18)F]FP)-TOCA: correlation between uptake, size and arterial perfusion in somatostatin receptor positive lesions. AB - PURPOSE: Somatostatin receptor (sstr) positive tumours vary widely in uptake of radiolabelled somatostatin (sst) analogues. This study determinates variability in lesion uptake of the glycosylated sst analogon N(alpha)-(1-deoxy-D-fructosyl) N(epsilon)-(2-[(18)F]fluoropropionyl)-Lys(0)-Tyr(3)-octreotate (Gluc Lys([(18)F]FP)-TOCA) and correlates it with lesion size and arterial perfusion as measured on computed tomography (CT). METHODS: Ten patients with metastasized neuroendocrine carcinomas were investigated with positron emission tomography PET/CT (Biograph 16, Siemens, Germany). Lesion standardized uptake values (SUVs) were determined at approximately 50 min post tracer injection according to a 60% isocontour volume of interest around each lesion. Lesion size and enhancement in the arterial phase (hounsfield units, HUs) were derived from CT. RESULTS: 114 lesions in the upper abdomen had a correlate on both, PET and CT. Variability in lesion SUVs was high (SUV(mean) 22 +/- 13). Intraindividually, there was a sigmoid positive correlation between lesion SUV and lesion diameter indicating partial volume effects. Residual variability in lesions > or =3 cm (> or =2.5 cm) ranged down to about half (third) of the maximum lesion uptake and remained unexplained by partial volume effects. No correlation with measured HU in the arterial phase was found, neither intraindividually nor interindividually. CONCLUSION: Partial volume effects were a major source of intraindividual variability in tumour tracer uptake. Lesions below 2.5 to 3 cm should thus be used with caution when performing dose calculations. In larger lesions residual variability in uptake must be considered; it may be due to variable sstr2 expression on the tumours' cell surfaces. PMID- 17912525 TI - Multicentre assessment and monitored use of [(18)F]FDG-PET in oncology: the Spanish experience. AB - PURPOSE: The aim of this study was to evaluate the diagnostic effectiveness of [(18)F]FDG-PET in oncological diseases and to assess its clinical utility and impact (on the clinical and therapeutic management of these patients). This health technology assessment was performed in Spain, using the monitored use (MU) procedure. METHODS: A multicentre and prospective follow-up study was performed in a non-consecutive sample of oncological patients who were examined with PET and other conventional diagnostic tests. A protocol for this MU method (PET-MU protocol) was developed, including the three forms used to collect all the information. Enrollment of new patients began in June 2002 and continued until August 2004. A descriptive analysis and an evaluation of the diagnostic effectiveness of FDG-PET were performed. RESULTS: The study population comprised 2,824 oncological patients (the third form relating to follow-up was completed for only 967 of these patients) from 100 Spanish hospitals and 16 PET centres. Seventy-nine percent of cases met the clinical requirements of the PET-MU protocol. Global diagnostic parameters of PET performance and their 95% CI values were as follows: sensitivity 86% (82-89%), specificity 83% (79-86%), positive and negative predictive values 87% (83-90%) and 82% (77-85%) respectively, diagnostic accuracy 84% (82-87%) and diagnostic odds ratio 28.75 (19.75-41.84). PET detected unsuspected new lesions in 39% of patients and avoided other unnecessary diagnostic techniques and treatments in 69% of cases. In 88% of cases, PET was considered useful by the physicians who asked for the PET tests (it was deemed decisive in 30% and very useful in almost 37%). CONCLUSION: This PET-MU study has confirmed the high diagnostic effectiveness of FDG-PET for oncological indications and demonstrates that it has a great influence on the clinical and therapeutic management of patients. PMID- 17912526 TI - Antiproliferative effects of rapamycin as a single agent and in combination with carboplatin and paclitaxel in head and neck cancer cell lines. AB - PURPOSE: Recent data suggested that combining targeted therapies with chemotherapy may counteract drug resistance. Activation of the PI3K/AKT/mTOR pathway downstream to kinase receptors, such as EGFR, was found in 57-81% of head and neck squamous cell carcinoma (HNSCC), and was eventually associated with a loss of PTEN function. mTOR was shown to modulate cell proliferation, apoptosis, invasion, and angiogenesis. This study aimed to evaluate molecular and cellular effects of rapamycin in a panel of cell lines either as single agent or in combination with cytotoxics commonly used in HNSCC. METHODS: Antiproliferative effects of rapamycin, carboplatin, and paclitaxel were evaluated in a panel of three HNSCC cell lines (SCC61, SQ20B and HEP2). Cells were exposed to rapamycin for 48 h, to carboplatin for 48 h, or to paclitaxel for 24 h. Antiproliferative effects of simultaneous and sequential rapamycin-based combinations were studied using MTT assay and median effect plot analysis. Cell cycle effects were analysed using flow cytometry. RESULTS: Rapamycin induced concentration dependent antiproliferative effects in HNSCC cell lines with IC(50) of 5 +/- 1, 12 +/- 2 and 20 +/- 2 microM in SCC61, SQ20B, and HEP2 cells, respectively. Higher antiproliferative effects were observed in SCC61 cells overexpressing NOXA and cyclin D1 than in HEP2 that overexpressed MDR1 and BCL2. In our panel, antiproliferative effects of rapamycin were associated with G0/G1 cell cycle accumulation and apoptosis induction, at concentrations ranging 3-30 microM. Combinations of rapamycin with paclitaxel and carboplatin displayed synergistic and additive effects. Synergistic effects were observed with paclitaxel in SQ20B and HEP2 cells and with carboplatin in SQ20B cells, when cells were exposed to cytotoxics prior to rapamycin. CONCLUSION: Our results show that rapamycin displays antiproliferative effects and induces apoptosis in HNSCC cell lines, cellular effects being more potent in cells that do not express BCL2 and MDR1. Additive and synergistic effects were observed when rapamycin was combined with carboplatin and paclitaxel. PMID- 17912527 TI - Height gain of vertebral bodies and stabilization of vertebral geometry over one year after vertebroplasty of osteoporotic vertebral fractures. AB - The height gain of vertebral bodies after vertebroplasty and geometrical stability was evaluated over a one-year period. Osteoporotic fractures were treated with vertebroplasty. The vertebral geometry and disc spaces were analysed using reformatted computed tomography (CT) images: heights of the anterior, posterior, and lateral vertebral walls, disc spaces, endplate angles, and minimal endplate distances. Vertebrae were assigned to group I [severe compression (anterior height/posterior height) <0.75] and group II (moderate compression index >0.75). A total of 102 vertebral bodies in 40 patients (12 men, 28 women, age 70.3 +/- 9.5) were treated with vertebroplasty and prospectively followed for 12 months. Group I showed a greater benefit compared with group II with respect to anterior height gain (+2.1 +/- 1.9 vs +0.7 +/- 1.6 mm, P < 0.001), reduction of endplate angle (-3.6 +/- 4.2 vs -0.8 +/- 2.3 degrees , P < 0.001), and compression index (+0.09 +/- 0.11 vs +0.01 +/- 0.06, P < 0.001). At one-year follow-up, group I demonstrated preserved anterior height gain (+1.5 +/- 2.8 mm, P < 0.015) and improved endplate angle (-3.4 +/- 4.9 degrees , P < 0.001). In group II, the vertebral heights returned to and were fixed at the pre interventional levels. Vertebroplasty provided vertebral height gain over one year, particularly in cases with severe compression. Vertebrae with moderate compression were fixed and stabilized at the pre-treatment level over one year. PMID- 17912528 TI - A prospective randomized comparative study on the use of ventral subcutaneous flap to prevent fistulas in the Snodgrass repair for distal hypospadias. AB - We evaluated the importance of urethral coverage using vascularized subcutaneous ventral flaps for the prevention of fistulas in patients undergoing distal hypospadias repair. Our prospective study included 130 patients, aged 9 months to 12 years, who underwent distal hypospadias repair using tubularized incised plate urethroplasty (TIPU), from January 2001 through January 2006. Patients were assigned to one of two groups by a computer-generated random selection: 65 patients underwent non-covered urethroplasty (NCU group); another group of 65 patients underwent covered urethroplasty (CU group) with a vascularized subcutaneous ventral flap. The results were evaluated by two pediatric surgeons unaware of the type of treatment each patient had undergone. Successful results were achieved in 99/130 patients (76.2%). We recorded 31 (23.8%) post-operative complications: 20 patients presented with a urethrocutaneous fistula (15 patients in the NCU group and 5 in the CU group); five with urethral stenosis (3 in the NCU and 2 in the CU group); and six with skin dehiscence of the preputioplasty (3 patients in each group). We analyzed the results using the chi2 test and the only statistically significant difference between the two groups (p < 0.05) was in terms of incidence of fistulas. Urethrocutaneous fistulas seem to be the most frequent complication of distal hypospadias after TIPU repair. Urethral coverage should be part of the Snodgrass procedure because it significantly reduces the formation of fistulas. A well-vascularized subcutaneous ventral flap represents, in our experience, a simple and optimal choice for the prevention of fistulas. PMID- 17912529 TI - Role of FGFR3 in urothelial cell carcinoma: biomarker and potential therapeutic target. AB - Although non-invasive bladder tumours (pTa) are the most common group of bladder tumours at presentation, there has until recently been relatively little information on their molecular biology. Thus it was of great interest when mutations in the FGF receptor 3 (FGFR3) were identified in bladder tumours and it became apparent that these were most common in tumours of low grade and stage. Since the initial description of activating mutations of FGFR3, there have been numerous studies confirming the frequency and spectrum of these mutations in bladder cancers of all grades and stages. Mutation screening techniques have evolved and improved. FGFR3 mutation has been assessed as a predictive biomarker in tumour tissues and as a diagnostic biomarker in urine. Efforts have been made to understand the function of FGFR3 in urothelial and other cells. Although our understanding of FGFR3 function is incomplete, it is already apparent that this may represent an important therapeutic target not only in non-invasive bladder cancer but also in a significant number of invasive tumours. This review summarises the current state of knowledge of this interesting receptor in urothelial carcinoma (UC). PMID- 17912530 TI - Subcutaneous tunneling of the temporary testing electrode significantly improves the success rate of subchronic sacral nerve modulation (SNM). AB - Sacral nerve modulation (SNM) is an effective way to treat non-neurogenic dysfunctions of pelvic organs. For over 20 years, this technique has been used for the treatment of overactive bladder, urinary retention, pelvic pain and even more recently, fecal incontinence and constipation. The objective of the study is to improve the fixation of the temporary testing electrode (TTE) in order to obtain more reliable results in the testing phase which should lead to have a comparable success rate as the two-stage implant for a chronic implant. Twenty eight patients (ratio of sex women:men = 3:1; with overactive bladder, urinary retention, pelvic pain syndrome and fecal incontinence) were evaluated by the modified temporary test electrode (TTE) placement. With the subcutaneous tunneling technique (mean time of evaluation 8.3 days), it is possible to perform percutaneous nerve evaluation (PNE) more effectively with an objective, reliable and less expensive outcome prior to the implantation of the implantable sacral nerve stimulator in almost 80% of the evaluated patients. Because the costs of therapy are not covered by health insurance in all countries, there is a need for an effective and inexpensive way to test and select patients appropriately. The tunneled TTE maintains its place for consistent amplitude during the entire test duration. The modification of placing the TTE produces repayable results. This technique can be performed on an outpatient basis to evaluate sacral nerve modulation as an early treatment option for non-dysfunctions of pelvic organs before they are forwarded to a specialized center for a chronic SNM implantation. PMID- 17912531 TI - [Long-term results after pars plana vitrectomy with 25 gauge technique]. AB - BACKGROUND: The 25-gauge technique of pars plana vitrectomy appears to be a very suitable method, especially for patients with pathological epiretinal alterations of the macula. However, the procedure has been criticized for insufficient impermeability with an increased risk of endophthalmitis and that the flexibility of instruments is too high. METHOD: Between 2002 and 2006, 625 eyes from 620 patients were operated on using the 25-gauge technique. Epiretinal membranes in different stages had been diagnosed in all patients. The operations were performed by only one surgeon. RESULTS: The epiretinal membranes were successfully removed in all patients and 329 eyes were analyzed with long-term follow up over 3.1 years. The mean improvement in visual acuity before and after surgery was -0.41 in LogMAR. One week postoperatively normal IOP was observed in all cases. The mean preoperative IOP was 17 mmHg and 8 mmHg 1 day after surgery. In nine patients with postoperative hypotony and choroidal detachment an additional suture was required and seven patients developed a retinal detachment. Endophthalmitis was not observed in any of the patients during the follow-up period. CONCLUSIONS: The 25-gauge PPV technique appears to be effective and safe for the treatment of epiretinal membranes. The operation has low complication rates with respect to endophthalmitis or retinal detachment. The procedure has recently been further improved by using more stable instruments and better lighting. PMID- 17912532 TI - [Application of wavefront analysis in clinical and scientific settings. From irregular astigmatism to aberrations of a higher order--Part I: Basic principles]. AB - In recent years, wavefront analysis has emerged from a pure laboratory application to an ophthalmological diagnostic method. This development was promoted mainly by the widespread use of wavefont-guided LASIK. However, aberrometry is still not a common diagnostic technique and the results of measurements are difficult to understand and to interpret for many ophthalmologists. The first part of this 2-part series summarizes the basics of wavefront errors, aberrometry and wavefront analysis to give a comprehensive overview for the interested ophthalmologist and clinical scientist. The second part will review such findings that are relevant for the ophthalmological community and highlight current scientific applications. PMID- 17912533 TI - Acidity enhances the effectiveness of active chemical defensive secretions of sea hares, Aplysia californica, against spiny lobsters, Panulirus interruptus. AB - Sea hares such as Aplysia californica, gastropod molluscs lacking a protective shell, can release a purple cloud of chemicals when vigorously attacked by predators. This active chemical defense is composed of two glandular secretions, ink and opaline, both of which contain an array of compounds. This secretion defends sea hares against predators such as California spiny lobsters Panulirus interruptus via multiple mechanisms, one of which is phagomimicry, in which secretions containing feeding chemicals attract and distract predators toward the secretion and away from the sea hare. We show here that ink and opaline are highly acidic, both having a pH of approximately 5. We examined if the acidity of ink and opaline affects their phagomimetic properties. We tested behavioral and electrophysiological responses of chemoreceptor neurons in the olfactory and gustatory organs of P. interruptus, to ink and opaline of A. californica within their natural range of pH values, from approximately 5 to 8. Both behavioral and electrophysiological responses to ink and opaline were enhanced at low pH, and low pH alone accounted for most of this effect. Our data suggest that acidity enhances the phagomimetic chemical defense of sea hares. PMID- 17912534 TI - 3-Iodothyronamine: a novel hormone controlling the balance between glucose and lipid utilisation. AB - 3-Iodothyronamine is considered as a derivate of thyroid hormone as a result of enzymatic deiodination and decarboxylation. The physiological role of thyronamine (T1AM) is not known. The aim of this study was to analyze the metabolic response to T1AM in the Djungarian hamster Phodopus sungorus. We measured the influence of T1AM (50 mg/kg) on metabolic rate (VO(2)), body temperature (T (b)) and respiratory quotient (RQ) in this species and in BL/6 mice. T1AM treated hamsters as well as the mice showed a rapid decrease in VO(2) and T (b), accompanied by a reduction of RQ from normal values of about approximately 0.9 to approximately 0.70 for several hours. This indicates that carbohydrate utilisation is blocked by the injection of T1AM and that metabolic pathways are rerouted from carbohydrate to lipid utilisation in response to T1AM. This assumption was further supported by the observation that the treatment of T1AM caused ketonuria and a significant loss of body fat. Our results indicate that T1AM has the potential to control the balance between glucose and lipid utilisation in vivo. PMID- 17912535 TI - Unstable upper pediatric cervical spine injuries: report of 28 cases and review of the literature. AB - OBJECTIVE: Traumatic lesions of upper cervical spine are rare in children. To evaluate their experience with this lesions and factors affecting outcome, authors conducted a retrospective study of 28 cases of upper pediatric cervical spine injuries treated in the last 25 years. MATERIALS AND METHODS: To help in treatment of these lesions, we studied our series and reviewed the literature. Patients were divided into three groups: 0 to 2, 3 to 8, and 9 to 16 years, and managed according to status at presentation and type of injury. RESULTS: Seven patients were managed surgically and 21 nonsurgically (3 halo braces, 18 hard collars or molded braces). Patients in the younger age group sustained more neurological injuries than the others. In group 1, 33% present a fracture/luxation of C0/C1 or odontoid. In group 2, 80% had sustained fracture/luxations of C2. In group 3, 60% present odontoid fractures. At late follow-up review, solid fusions were demonstrated in all patients. Neurological deterioration occur in six patients. The mortality rate was 12%. Compared with other authors' report, incidence of this lesions increased but not the number of those managed surgically. CONCLUSIONS: Management must be tailored to the patient's age, neurological status, type, and level of injury. Compared with other author's experience, fusion and instrumentation procedures were used less frequently. Signs of medullary compression, significant spine deformation, dynamic instability, and age higher than 8 years are the criteria for surgery. The criteria for instability in children are different from those used in adults because the residual spinal growth is a major concern. The best treatment is therefore preventive. PMID- 17912536 TI - Health economic evaluation of ferucarbotran-enhanced MRI in the diagnosis of liver metastases in colorectal cancer patients. AB - PURPOSE: The objective of our study was to analyze the health economic impact of ferucarbotran-enhanced magnetic resonance imaging (MRI) in the diagnosis of hepatic colorectal cancer metastases based on observed changes in medical management. MATERIALS AND METHODS: A decision tree simulating a patient's medical management was designed, comparing two scenarios: contrast-enhanced spiral computed tomography-based vs ferucarbotran-enhanced MRI-based (Resovist, Bayer Schering Pharma AG, Germany) diagnosis. A clinical trial in patients with presumed liver metastases (n=36) provided data on clinical decisions regarding the medical management options in relation to diagnostic outcomes: resection, chemotherapy, or best supportive care. A "gold standard" was established afterward, combining all the available clinical, imaging, laboratory, and pathology findings. A multidisciplinary panel formed by a hepatologist, a liver surgeon, and an interventional radiologist decided on the recommended medical management for each patient. Costs of medical resources associated with each management option (all expressed in Euro) were obtained from the public health insurance (average European values). Life expectancies for the different options were obtained from literature. RESULTS: Despite an initial extra cost of 338 Euro, a significant net saving of 1,443 Euro was obtained with ferucarbotran enhanced MRI mainly because of avoiding unnecessary surgery. There was no significant difference in the predicted life expectancy between both arms, despite the large difference in medical decision. CONCLUSION: In this comparative medical decision analysis, it was shown that ferucarbotran-enhanced MRI has the potential to improve medical management and save health care costs. PMID- 17912537 TI - p73 expression in medulloblastoma: TAp73/DeltaNp73 transcript detection and possible association of p73alpha/DeltaNp73 immunoreactivity with survival. AB - The human p73 protein is essential for normal morphogenesis and maintenance of neural tissue. Recently, several TP73 transcripts have been revealed in medulloblastoma (MB), the most common malignant brain tumor in children. Here, we performed immunohistochemical analysis on 29 MB specimens using anti-p73alpha and anti-DeltaNp73 antibodies. Real-time PCR quantification was performed to assess TAp73 and DeltaNp73 transcripts in a subset of 13 MB samples. Normal cerebellar tissues and RNA were used for comparison. Pilot clinical-pathological correlations were also provided. We report significant differences for TAp73 and DeltaNp73 mRNA expression between tumor tissues and reference (P = 0.013, P = 0.028). Immunohistochemically, 52 and 29% MB samples were positive for p73alpha and DeltaNp73, respectively. p73alpha expression was found to be in both the nucleus and cytoplasm, whereas DeltaNp73 was localized predominantly in the cytoplasm. In normal cerebellum, positive staining for p73alpha and DeltaNp73 was observed in the Purkinje cells of newborns, not adult samples, which supports the developmental role of TP73 during organogenesis of the human cerebellum. Survival analysis has shown negative relationship of DeltaNp73-immunoreactivity with overall survival (OS) and event free survival (EFS) (P = 0.026 and P = 0.127, respectively). For p73alpha-positive cases, the negative trend in OS (P = 0.149) and EFS (P = 0.216) was also apparent. Our results indicate the involvement of p73 protein in MB tumorigenesis and define TP73 as a potential prognostic and therapeutic target for medulloblastoma. PMID- 17912539 TI - Chronic post-traumatic radial head dislocation associated with dissociation of distal radio-ulnar joint: a case report. AB - We present an unusual case of an isolated interosseous membrane disruption of the forearm without any fracture pattern. Dislocation of both radial head and distal radio-ulnar joint was presented. Open reduction of the radial head with radial neck shortening osteotomy was performed. PMID- 17912540 TI - Persistent subumbilical discharge associated with actinomycosis caused by intrauterine contraceptive device: a case report. AB - Intrauterine devices are a very effective form of contraception used worldwide for more than 2 centuries. One of the serious, but rare, complications of intrauterine contraception is perforation through the uterine wall into the pelvic or abdominal cavity. In this report we describe, persistent periumbilical discharge associated actinomycosis caused by migration of a copper intrauterine contraceptive device. To our knowledge, this is the first report of persistent periumbilical discharge caused by migration of a copper intrauterine contraceptive device. We recommend consideration of this minor possibility during evaluation of women with chronic abdominal discharge. PMID- 17912541 TI - Massive ovarian edema in pregnancy after ovulation induction using clomiphene citrate. AB - BACKGROUND: Massive ovarian edema is a benign enlargement of the ovary caused by accumulation of fluid occurring mainly in young women. Most cases are thought to result from venous and lymphatic obstruction. CASE REPORT: We treated a 40-year old multiparous pregnant woman with massive ovarian edema who had been received clomiphene citrate. She was admitted at 13th week of pregnancy for acute pelvic pain. Left oophorectomy was performed, and pathologic examination disclosed massive ovarian edema. Our report is the first case of massive ovarian edema with pregnancy after ovulation induction using clomiphene citrate. PMID- 17912538 TI - Gray matter injury associated with periventricular leukomalacia in the premature infant. AB - Neuroimaging studies indicate reduced volumes of certain gray matter regions in survivors of prematurity with periventricular leukomalacia (PVL). We hypothesized that subacute and/or chronic gray matter lesions are increased in incidence and severity in PVL cases compared to non-PVL cases at autopsy. Forty-one cases of premature infants were divided based on cerebral white matter histology: PVL (n = 17) with cerebral white matter gliosis and focal periventricular necrosis; diffuse white matter gliosis (DWMG) (n = 17) without necrosis; and "Negative" group (n = 7) with no abnormalities. Neuronal loss was found almost exclusively in PVL, with significantly increased incidence and severity in the thalamus (38%), globus pallidus (33%), and cerebellar dentate nucleus (29%) compared to DWMG cases. The incidence of gliosis was significantly increased in PVL compared to DWMG cases in the deep gray nuclei (thalamus/basal ganglia; 50-60% of PVL cases), and basis pontis (100% of PVL cases). Thalamic and basal ganglionic lesions occur almost exclusively in infants with PVL. Gray matter lesions occur in a third or more of PVL cases suggesting that white matter injury generally does not occur in isolation, and that the term "perinatal panencephalopathy" may better describe the scope of the neuropathology. PMID- 17912543 TI - Antifungal efficacy of voriconazole, itraconazole and amphotericin b in experimental fusarium solani keratitis. AB - PURPOSE: To evaluate the efficacy of topical voriconazole in an experimental rabbit model of Fusarium keratitis. METHODS: Fungal keratitis was induced in the right eyes of 24 New Zealand rabbits. 8.6 x 10(3) CFU/0.1 ml F.solani spore suspension was injected midstromally into the central cornea. Group 1 received topical amphotericin B 0.15%, group 2 received topical itraconazole 1% and group 3 received topical voriconazole 1% hourly between 08:00 to 22:00 on days 1 and 2; 4 times daily on days 3-5. Control group received topical balanced salt solution at identical intervals. The eyes were examined clinically with a scoring system before treatment (day 0), on day 3 and on day 5. Cultures were taken from the lesion by scraping at the end of the treatment. Clinical scores and microbiologic results were analyzed statistically. RESULTS: In the control group, keratitis progressed clinically and colony level was 2 x 10(3) CFU at day 5. In all treatment groups, progression of keratitis was inhibited clinically. Culture was sterile in the group receiving amphotericin B. Colony level was 0.3 x 10(2) CFU in the itraconazole group and 2 x 10(2) CFU in the voriconazole group at day 5. CONCLUSION: Progression of keratitis was inhibited clinically in all treatment groups. Colony level decreased significantly in all treatment groups. As a result, itraconazole 1% and voriconazole 1% were found to be effective in Fusarium keratitis clinically and microbiologically, although their activity was not as effective as amphotericin B 0.15%. PMID- 17912544 TI - Chained learning architectures in a simple closed-loop behavioural context. AB - OBJECTIVE: Living creatures can learn or improve their behaviour by temporally correlating sensor cues where near-senses (e.g., touch, taste) follow after far senses (vision, smell). Such type of learning is related to classical and/or operant conditioning. Algorithmically all these approaches are very simple and consist of single learning unit. The current study is trying to solve this problem focusing on chained learning architectures in a simple closed-loop behavioural context. METHODS: We applied temporal sequence learning (Porr B and Worgotter F 2006) in a closed-loop behavioural system where a driving robot learns to follow a line. Here for the first time we introduced two types of chained learning architectures named linear chain and honeycomb chain. We analyzed such architectures in an open and closed-loop context and compared them to the simple learning unit. CONCLUSIONS: By implementing two types of simple chained learning architectures we have demonstrated that stable behaviour can also be obtained in such architectures. Results also suggest that chained architectures can be employed and better behavioural performance can be obtained compared to simple architectures in cases where we have sparse inputs in time and learning normally fails because of weak correlations. PMID- 17912545 TI - Red and green aequorins for simultaneous monitoring of Ca2+ signals from two different organelles. AB - Simultaneous control of different functions by calcium signals is possible because of subcellular compartmentalization. Targeted chemiluminescent Ca2+ probes, such as aequorins (AEQs) are optimal for detecting signals originating in different subcellular domains, but imaging is difficult because of low photon yield causing poor spatiotemporal resolution. To overcome this problem, we have co-expressed two spectrally distinct AEQs in different subcellular locations within the same cells. Seven chimeric proteins containing either green- or red emitting AEQs, with different targeting sequences and Ca2+ affinities, have been designed and tested. We show here evidence for physical and functional independence of the different probes. Cytosolic Ca2+ signals were mirrored in the nucleus, but amplified inside mitochondria and endoplasmic reticulum, and had different time courses in the various locations. Our results demonstrate that these novel tools permit simultaneous and independent monitoring of [Ca2+] in different subcellular domains of the same cell. PMID- 17912546 TI - The Wilms' tumor suppressor WT1 is associated with melanoma proliferation. AB - Expression of the Wilms' tumor suppressor WT1 has been demonstrated in a variety of tumors and tumor cell lines, e.g., in breast cancer and melanoma cell lines. Its role is controversial, with evidence for both tumor-promoting and tumor suppressing activities. In this paper, we show that WT1 is expressed in malignant melanoma in >80% of the tumor cells, but not in normal skin or benign melanocytic nevi in vivo. We detected an unusual shift of WT1 isoforms towards WT1(+17AA/+KTS) in melanoma. WT1 shared an overlapping expression with proliferating nuclear cell antigen and with Nestin and Zyxin, which are involved in melanoma cell proliferation. To investigate whether WT1 is directly involved in melanoma cell proliferation, we made use of an RNAi approach in vitro. WT1 silencing significantly reduced the expression of Nestin and Zyxin and resulted in inhibition of melanoma cell proliferation as determined by a reduced BrdU incorporation. These findings suggest a direct role of WT1 in melanoma proliferation, which might be mediated via Nestin and Zyxin. Furthermore, expression of WT1 in vivo clearly discriminates between benign acquired nevi and malignant melanomas and appears to be correlated with melanocytic atypia and malignancy. PMID- 17912547 TI - A refined radio-telemetry technique to monitor right ventricle or pulmonary artery pressures in rats: a useful tool in pulmonary hypertension research. AB - Implantable radio-telemetry methodology, allowing for continuous recording of pulmonary haemodynamics, has previously been used to assess effects of therapy on development and treatment of pulmonary hypertension. In the original procedure, rats were subjected to invasive thoracic surgery, which imposes significant stress that may disturb critical aspects of the cardiovascular system and delay recovery. In the present study, we describe and compare the original trans thoracic approach with a new, simpler trans-diaphragm approach for catheter placement, which avoids the need for surgical invasion of the thorax. Satisfactory overall success rates up to 75% were achieved in both approaches, and right ventricular pressures and heart and respiratory rates normalised within 2 weeks. However, recovery was significantly faster in trans-diaphragm than in trans-thoracic operated animals (6.4+/-0.5 vs 9.5+/-1.1 days, respectively; p<0.05). Stable right ventricular pressures were recorded for more than 4 months, and pressure changes, induced by monocrotaline or pulmonary embolisms, were readily detected. The data demonstrate that right ventricular telemetry is a practicable procedure and a useful tool in pulmonary hypertension research in rats, especially when used in combination with echocardiography. We conclude that the described trans-diaphragm approach should be considered as the method of choice, for it is less invasive and simpler to perform. PMID- 17912548 TI - Isolated vasculitis of the female genital tract: a case series and review of literature. AB - To provide a clinicopathologic review of vasculitis confined to the female genital tract, we describe three cases, and we searched PubMed from 1965 to 2006 with analysis of all relevant articles. We identified 118 additional cases in the literature of whom 108 had isolated necrotizing vasculitis similar to classical polyarteritis nodosa (PAN-type), and 10 presented isolated giant cell arteritis (GCA-type) of the female genital tract. In most cases, arteritis was discovered surprisingly. The mean age of these patients was 48.6 years for the PAN-type and 64.1 for the GCA-type. Vasculitis affected a single organ in 88 (81.5%) cases of which 71 (65.7%) involved the cervix in the PAN-type, whereas it was limited in the myometrium in five (50%) cases in the GCA-type. There was no progression to a systemic vasculitis in 99.1% of the cases. A comparable favorable outcome was reported after surgery in all cases with a mean follow-up of 40.8 months. We conclude that isolated vasculitis of the female genital tract is a distinct condition, with two different patterns but a similar favorable outcome after resection. Its knowledge is needed to avoid aggressive evaluation and therapy. PMID- 17912549 TI - The effect of vestibular nerve section on the expression of the hyaluronan in the frog, Rana esculenta. AB - Following postganglionic lesion of the eighth cranial nerve, the changes in the expression of hyaluronan (HA), one of the extracellular matrix macromolecules, were examined in the medial (MVN) and lateral (LVN) vestibular nuclei and in the entry or transitional zone (TZ) of the nerve in the frog. HA was detected in different survival times by using a specific biotinylated hyaluronan-binding probe. HA expression was defined by the area-integrated optical density (AIOD), calculated from pixel intensities of digitally captured images. During the first postoperative days the perineuronal net (PN), a HA-rich area around the neurons, was not distinguishable from the surrounding neuropil in the MVN and LVN, characterized by a bilateral drop of AIOD specifically on the operated side. From postoperative day 14 onwards AIOD increased whilst the PN reorganized. In contrast, the AIOD wobbled up and down bilaterally without any trend in the TZ. Statistical analysis indicated that AIOD changes in the structures studied ran parallel bilaterally presumably because of the operation. Our results demonstrated for the first time that (1) the lesion of the eighth cranial nerve is accompanied by the modification of AIOD reflected HA expression in the MVN, LVN and TZ, (2) different tendencies exist in the time course of AIOD in the structures studied and (3) these tendencies are similar on the intact and operated sides. Our findings may suggest an area dependent molecular mechanism of HA in the restoration of vestibular function. PMID- 17912550 TI - What's new in metabolic and genetic hypoglycaemias: diagnosis and management. AB - Hypoglycaemia in children can be a life-threatening situation that needs to be assessed rigorously in order to treat efficiently and avoid relapse that can be responsible for cerebral damage. The diagnosis of impairment in glucose homeostasis requires the knowledge of the mechanisms regulating blood glucose concentration. The clinical history and presentation, when available, especially the timing of hypoglycaemia with respect to the last meal and some simple clinical and biological tests may allow diagnosing the vast majority of patients presenting with hypoglycaemia. Recently, new metabolic and endocrinologic genetic causes of hypoglycaemia have been identified that may give new insight to the complex mechanisms of glucose regulation and thus contribute to the discovery of new genes regulating glucose homeostasis. New diagnostic tests such as the 18 fluoro-Dopa PET-scan have also been recently developed. PMID- 17912551 TI - A case of herpes simplex encephalitis revealed by decompressive craniectomy. AB - A 15-year-old girl was referred to our hospital due to fever, headache, and vomiting of 7 days duration and focal motor convulsion at the day of referral. Her clinical signs and cerebral imaging findings were found to be compatible with herpes simplex encephalitis. In spite of prompt acyclovir administration, her consciousness deteriorated gradually. Emergent cranial magnetic resonance imaging demonstrated a shift of midline intracranial structures. Decompressive surgery resulted in partial improvement in the shift of midline intracranial structures and potentially saved the patient's life. This case report stresses the importance of proper management of increased intracranial pressure in patients with herpes simplex encephalitis. PMID- 17912552 TI - Cross-sectional surveys and subtype classification of human Blastocystis isolates from four epidemiological settings in China. AB - The prevalence and geographical distribution of the intestinal protozoa Blastocystis in humans across China is unknown, and the relative importance of different subtypes has yet to be investigated. We assessed the community prevalence and relative frequencies of different Blastocystis subtypes in four epidemiological settings in China, i.e., Shanghai municipality, Yongjia county (Zhejiang province), Eryuan county, and Menghai county (both Yunnan province). Blastocystis infection was detected with the culture method, and the subtype was identified with polymerase chain reaction using a set of subtype-specific primers. The prevalence at the four study settings was 1.9, 5.9, 18.4, and 32.6%, respectively. People aged greater than or equal to 60 years had a higher prevalence in the former two settings, Shanghai and Yongjia, whereas the highest infection rate was found among individuals aged 10-17 years in the latter two settings, Eryuan and Menghai. A higher prevalence was found in men in the former two settings but in women in the latter two settings. Five different Blastocystis subtypes were identified from the 192 isolates. Subtype 3 was the predominant type, followed by subtype 1. In conclusion, the epidemiology of Blastocystis varies across China. PMID- 17912553 TI - Retinal stem cells: promising candidates for retina transplantation. AB - Stem cell transplantation is widely considered as a promising therapeutic approach for photoreceptor degeneration, one of the major causes of blindness. In this review, we focus on the biology of retinal stem cells (RSCs) and progenitor cells (RPCs) isolated from fetal, postnatal, and adult animals, with emphasis on those from rodents and humans. We discuss the origin of RSCs/RPCs, the markers expressed by these cells and the conditions for the isolation, culture, and differentiation of these cells in vitro or in vivo by induction with exogenous stimulation. PMID- 17912554 TI - Familial hematuria. AB - Hematuria is a common presenting complaint in pediatric nephrology clinics and often has a familial basis. This teaching article provides an overview of causes, diagnosis, and management of the major forms of familial hematuria, Alport syndrome, and thin basement membrane nephropathy. PMID- 17912555 TI - Chronic recurrent multifocal osteomyelitis presenting as acute scoliosis: a case report and review of literature. AB - Chronic relapsing multifocal osteomyelitis (CRMO) is a rare condition. It commonly affects the clavicle and pelvis. Rarely it can affect the spine. Spinal deformity due to CRMO is rare. We report a case of acute scoliosis due to CRMO. A 10-year-old girl with CRMO presented with acute painful scoliosis of her spine. She was neurologically intact. Imaging suggested a neoplastic process involving T10, L2 and L3. Further imaging and subsequent biopsy was performed and a diagnosis of CRMO was established. Spinal involvement with deformity is uncommon. It is commonly misdiagnosed as infection or a neoplasm and unnecessary aggressive surgical and antibiotic therapy instituted. A high index of suspicion is needed to diagnose this disease and thus manage it appropriately. This patient with a previously normal spine had a long right sided thoracic scoliosis. We think that the particular pattern of scoliosis was a protective mechanism to offload the right sided T10 vertebral pedicle. Prognosis is generally good although the disease can relapse and remit over many years. At 9 months follow up, the lesions were resolving and the deformity had resolved. CRMO presenting as acute scoliosis is rare and to our knowledge this is the second recognised case in the reported world literature. PMID- 17912556 TI - Radiographic evaluation of selective anterior thoracolumbar or lumbar fusion for adolescent idiopathic scoliosis. AB - According to Lenke classification of adolescent idiopathic scoliosis (AIS), patients with type 5 curve in which the structural major curve is thoracolumbar or lumbar curve with nonstructural proximal thoracic and main thoracic curves, could be surgically treated with selective anterior thoracolumbar or lumbar (TL/L) fusion. This study retrospectively analyzed the radiographies of selective anterior TL/L fusion in 35 cases of AIS with Lenke type 5 curve. Segmental fixation with a single rigid rod through anterior thoracoabdominal approach was applied in all patients. Measurements of scoliosis curve in preoperative, immediate postoperative and follow-up radiographies were analyzed. The average follow up time was 36 months (24-42 months). The average preoperative Cobb angle of the TL/L curve was 45.6 degrees and improved into 9.7 degrees immediate postoperatively, with 79.7% curve correction. In addition, the minor thoracic curve decreased from 29.7 degrees preoperatively to 17.6 degrees postoperatively, with a spontaneous correction of 41.5%. During the follow-up, a loss of 4.6 degrees correction was found and the average Cobb angle of TL/L increased to 14.4 degrees . Also, the minor thoracic curve increased to average 20.1 degrees with a loss of 2.4 degrees correction. Trunk shift deteriorated slightly immediate postoperatively and improved at the follow-up. The lowest instrumented vertebra (LIV) tilt was improved significantly and maintained its results at the follow up. During the follow-up, the coronal disc angle immediately above the upper instrumented vertebra (UIVDA) and below the LIV (LIVDA) aggravated, while the sagittal contours of T5-T12 and T10-L2 were well maintained. The lumbar lordosis of L1-S1 and the sagittal Cobb angle of the instrumented segments were reduced slightly postoperatively and at the follow-up. There were no major complications or pseudarthrosis. The outcomes of this study show that selective anterior thoracolumbar or lumbar fusion with solid rod instrumentation is effective for surgical correction of AIS with Lenke type 5 curve. The TL/L curve, minor thoracic curve, and LIV title can be improved significantly, with good maintenance of sagittal contour. However, the UIVDA and LIVDA aggravate postoperatively when the trunk rebalances itself during follow-up. The degeneration of LIV disc warrants longer-term follow-up. PMID- 17912557 TI - The change in ratio of convex and concave lung volume in adolescent idiopathic scoliosis: a 3D CT scan based cross sectional study of effect of severity of curve on convex and concave lung volumes in 99 cases. AB - To study the effect of the degree of scoliosis, degree of hypokyphosis/lordosis and rotation of apical vertebra on individual lung volume (measured with CT scan) in asymptomatic adolescent idiopathic scoliosis (AIS) patients. Individual (right and left) lung volume, angle of kyphosis and rotation of apical vertebra, were measured in 77 asymptomatic AIS patients having right thoracic curve, using modern computed tomography (CT) scan. To compare, lung volumes were measured in 22 normal persons (control group). The ratio of "right to left lung volume (convex to concave side)" was obtained and compared among these groups. With increased Cobb's angle, ratio of convex to concave lung volume increased. For Cobb's angle more than 40 degrees, it was increased significantly (P = 0.0042). A significant degree of correlation was found between axial rotation angle of apical vertebra and right to left lung volume ratio (P = 0.0067, r = 0.271). A significant inverse correlation was found between the angle of kyphosis and right to left lung volume ratio, i.e., as the angle of kyphosis decreased the convex to concave lung volume ratio increased (P = 0.0109, r = -0.255). In asymptomatic, AIS patients, with increase in degree of curvature, and rotation of apical vertebra, the ratio of convex to concave side lung volume increases; indicating concave side lung volume is comparatively more affected (decreased) than convex side lung volume. On the other hand with decrease in the angle of kyphosis the convex to concave lung volume ratio increases indicating kyphotic angle has an inverse relation to convex to concave lung volume ratio. PMID- 17912558 TI - Radiographic measurement of the sagittal plane deformity in patients with osteoporotic spinal fractures evaluation of intrinsic error. AB - Cobb method has been shown to be the most reliable technique with a reasonable measurement error to determine the kyphosis in fresh fractures of young patients. However, measurement errors may be higher for elderly patients as it may be difficult to determine the landmarks due to osteopenia and the degenerative changes. The aim of this study is to investigate the intrinsic error for different techniques used in evaluation of local sagittal plane deformity caused by OVCF. Lateral X-rays of OVCF patients were randomly selected. Patient group was composed of 28 females and 7 males and the mean age was 62.7 (55-75) years. The kyphosis angle and the vertebral body height were analyzed to reveal the severity of sagittal plane deformity. Kyphotic deformity was measured by using four different techniques; and the vertebral body heights (VBH) were measured at three different points. The mean intra-observer agreement interval for kyphosis angle measurement techniques ranged from +/-7.1 to +/-9.3 degrees while it ranged from +/-4.5 to +/-6.5 mm for VBH measurement techniques. The mean interobserver agreement interval for kyphosis angle ranged from +/-8.2 to +/-11.1 degrees , while it was between +/-4.5 to +/-6.5 mm for vertebral body height measurement techniques. This study revealed that although the intra and interobserver agreement were similar for all techniques, they are still higher than expected. These high intervals for measurement errors should be taken into account when interpreting the results of correction in local sagittal plane deformities of OVCF patients after surgical procedures such as vertebral augmentation techniques. PMID- 17912559 TI - Multimodal intraoperative monitoring (MIOM) during 409 lumbosacral surgical procedures in 409 patients. AB - A prospective study on 409 patients who received multimodel intraoperative monitoring (MIOM) during lumbosacral surgical procedures between March 2000 and December 2005 was carried out. The objective of this study was to determine the sensitivity and specificity of MIOM techniques used to monitor conus medullaris, cauda equina and nerve root function during lumbosacral decompression surgery. MIOM has increasingly become important to monitor ascending and descending pathways, giving immediate feedback information regarding any neurological deficit during the decompression and stabilisation procedure in the lumbosacral region. Intraoperative spinal- and cortical-evoked potentials, combined with continuous EMG- and motor-evoked potentials of the muscles, were evaluated and compared with postoperative clinical neurological changes. A total of 409 consecutive patients with lumbosacral spinal stenosis with or without instability were monitored by MIOM during the entire surgical procedure. A total of 388 patients presented true-negative findings while two patients presented false negative and 1 patient false-positive findings. Eighteen patients presented true positive findings where neurological deficit after the operation was intraoperatively predicted. Of the 18 true-positive findings, 12 patients recovered completely; however, 6 patients recovered only partially. The sensitivity of MIOM applied during decompression and fusion surgery of the lumbosacral region was calculated as 90%, and the specificity was calculated as 99.7%. On the basis of the results of this study, MIOM is an effective method of monitoring the conus medullaris, cauda equina and nerve root function during surgery at the lumbosacral junctions and might reduce postoperative surgical related complications and therefore improve the long-term results. PMID- 17912560 TI - Novel flexible forceps for endoscopic transsphenoidal resection of pituitary tumors: technical report. AB - The endoscope has become an essential tool for transsphenoidal pituitary surgeries because of the panoramic view and the ability to visualize structures beyond the direct line of vision. However, the regular dedicated instruments for these surgeries sometimes do not reach the areas visualized by an angled endoscope, and instrument maneuverability is limited. To overcome or minimize these problems, we constructed a new flexible tumor forceps that can be manipulated to change shape after placing into the surgical fields via the endonasal or sublabial route, to reach any desirable location for tumor dissection and excision. We applied this newly developed instrument in endonasal endoscopic resection complementing microsurgical procedure in 20 cases of pituitary tumors. The flexible forceps was able to access sites where regular dedicated instruments for transsphenoidal pituitary surgeries could not readily reach despite endoscopic visualization. No complication was observed in the present series, confirming the safety of the newly designed flexible forceps. Our newly developed flexible forceps improves the instrument maneuverability of endoscopic transsphenoidal resection of pituitary tumors, especially those located laterally at the cavernous sinuses or with frontal extension. The use of this instrument makes the best use of endoscopic panoramic visualization for tumor removal. PMID- 17912561 TI - Retroperitoneal mesenchymal chondrosarcoma mimicking a large retroperitoneal sacral schwannoma. AB - We report a case of retroperitoneal chondrosarcoma in which preoperative radiological study induced to a possible diagnosis of sacral schwannoma. A 25 year-old woman was admitted to our neurosurgical institute for a progressive sciatic pain with gait difficulties with a sudden radicular deficit with right positive Lasegue's sign at 30 degrees, loss of Achilles reflex, sensory deficit on right S1 dermatome, and complete motor deficit of right plantar flexion. We performed an en bloc removal of the lesion via an anterior retroperitoneal approach. We believe that the treatment of choice is radical surgical excision of the tumor with complete en-bloc removal of the lesion; postoperative radiotherapy and chemotherapy should be valuated case by case. PMID- 17912562 TI - Treatment of hemodynamic insufficiency in chronic CCA occlusion using a short saphenous vein interposition graft: diagnostic and technical considerations. An illustrative case report. AB - Extracranial-intracranial bypass surgery has been shown to reverse hemodynamic insufficiency on the basis of steno-occlusive disease of the internal carotid artery (ICA) or middle cerebral artery. In contrast, chronic occlusion of the common carotid artery (CCA) without extracranial donor vessels affords alternative revascularization procedures as well as a more elaborate preoperative workup. This case is intended to illustrate the specific diagnostic approach and considerations as well as a beneficial treatment modality in the setting of pronounced hemodynamic insufficiency as a consequence of a CCA occlusion, in conjunction with contralateral CCA and ICA stenoses. A 61-year-old man complaining of new onset aphasia underwent vascular imaging that revealed a proximal occlusion of the left CCA with a concomitant patent proximal ICA on ultrasound. Functional cerebral blood flow measurement including Xenon-enhanced computer tomography showed corresponding chronic hemodynamic insufficiency of the left hemisphere. The patient received a modified revascularization procedure, where a saphenous vein was used as interposition graft between the subclavian artery and the left proximal ICA. Postoperatively, both clinical and morphological improvement were noted. Successful treatment of hemodynamic insufficiency because of chronic CCA occlusion necessitates a thorough preoperative workup and application of alternative revascularization strategies. PMID- 17912563 TI - Surgical treatment for choroid plexus tumors in the fourth ventricle: brain stem infiltration hinders total extirpation. AB - The significance of surgery for choroid plexus tumors is well established, but surgical resection of those in the fourth ventricle has not been evaluated. This study reviewed five consecutive patients with choroid plexus tumors in the fourth ventricle treated in our institute between 1996 and 2005, focusing on the factors that hindered total extirpation. Two cases were choroid plexus papillomas, and three cases were choroid plexus carcinomas. Preoperative T2-weighted magnetic resonance imaging showed a diffuse high-intensity lesion in the brain stem in four patients. Infiltration into the fourth ventricle floor was apparent in all five patients during surgery, which hindered total resection of the tumors without neurological deterioration. Intraoperative bleeding was well controlled in all five patients by cauterizing the feeding arteries at the early stage of surgery through the telovelar approach. Performance status was improved in all patients postoperatively. All patients with choroid plexus carcinomas underwent radiation therapy after the surgical removal. No patient suffered tumor progression within the follow-up of 24-129 months (mean 64 months). Total resection of choroid plexus tumors in the fourth ventricle is difficult because of invasion into the fourth ventricle floor. Adjuvant therapy for choroid plexus tumors with brain stem infiltration must be established. PMID- 17912564 TI - Expression of vascular endothelial growth factor in dura mater of patients with moyamoya disease. AB - Vascular endothelial growth factor (VEGF) has been found to be involved in vasculogenesis in different intracranial lesions. We investigated meningeal cellularity and VEGF expression in dura mater of patients with and without moyamoya disease. Nine dural specimens from nine cerebral hemispheres of seven patients with moyamoya disease and four control dural specimens from four non moyamoya patients were collected during surgery and investigated. Dural specimens were immunohistochemically stained with VEGF antibody, and then meningeal cellularity and VEGF expression in dural tissue were analyzed. The mean+/ standard error (SE) of total number of meningeal cells (meningeal cellularity) in dural tissue was 21.5+/-3.0 in the moyamoya disease patients, whereas it was 2.7+/-0.7 in control patients. The mean+/-SE of VEGF expression was 51.1+/-4.9% in the moyamoya disease patients, whereas it was 13.8+/-5.9% in control patients. The meningeal cellularity and VEGF expression were statistically significantly higher in the moyamoya group in comparison to control group (p<0.0001). Meningeal cellularity and VEGF expression are significantly increased in dura mater of the patients with moyamoya disease. PMID- 17912565 TI - Electronegative electroretinogram associated with topiramate toxicity and vitelliform maculopathy. AB - Topiramate is known to cause ocular side effects such as refractive changes and angle closure. We describe a patient with an electronegative electroretinogram (ERG) which may have been related to topiramate use. Electronegative ERG's have been associated with other drugs in humans as well as topiramate use in rabbits. However, this would be the first suggestion of causality in humans. PMID- 17912566 TI - Prediction of visual evoked potentials at any surface location from a set of three recording electrodes. AB - Purpose of this study was to introduce a mathematical model which allows the calculation of a source dipole as the origin of the evoked activity based on the data of three simultaneously recorded VEPs from different locations at the scalp surface to predict field potentials at any neighboring location and to validate this model by comparison with actual recordings. In 10 healthy subjects (25-38, mean 29 years) continuous VEPs were recorded via 96 channels. On the base of the recordings at the positions POz', O1' and O2', a source dipole vector was calculated for each time point of the recordings and VEP responses were back projected for any of the 96 electrode positions. Differences between the calculated and the actually recorded responses were quantified by coefficients of variation (CV). The prediction precision and response size depended on the distance between the electrode of the predicted response and the recording electrodes. After compensating this relationship using a polynomial function, the CV of the mean difference between calculated and recorded responses of the 10 subjects was 2.8 +/- 1.2%. In conclusion, the "Mini-Brainmapping" model can provide precise topographical information with minimal additional recording efforts with good reliability. The implementation of this method in a routine diagnostic setting as an "easy-to-do" procedure would allow to examine a large number of patients and normal subjects in a short time, and thus, a solid data base could be created to correlate well defined pathologies with topographical VEP changes. PMID- 17912569 TI - Vaccenic acid and cis-9,trans-11 CLA in the rumen and different tissues of pasture- and concentrate-fed beef cattle. AB - The objective of present study was the comparison of trans-11 18:1 (VA) and cis 9,trans-11 CLA concentrations in the rumen and different tissues in beef cattle, and to examine the diet and breed effects on the compound concentration and deposition. Sixty-four German Holstein and German Simmental bulls were randomly assigned to two dietary treatments, based on concentrate or pasture. The concentration of cis-9,trans-11 CLA and VA in rumen, duodenal digesta and different tissues was determined by gas chromatography. The results showed that pasture relative to concentrate feeding significantly increased the concentration of VA in duodenal digesta, plasma and erythrocyte phospholipids. Pasture-based feeding resulted in a significant enrichment of cis-9,trans-11 CLA in plasma lipids and erythrocyte phospholipids, but not in rumen and duodenal digesta, compared to concentrate-fed diet. Diet did not affect the cis-9,trans-11 CLA concentrations (mg/100 g fresh tissue) in semitendinosus muscle and subcutaneous fat. There was a breed effect on the deposition of cis-9,trans-11 CLA in longissimus muscle with lower concentration in pasture-fed German Simmental bulls compared to concentrate-fed bulls. However, pasture feeding significantly increased both, the VA and cis-9,trans-11 CLA concentrations in liver and heart tissues. Both diet and breed effects on Delta(9)-desaturase index was observed in muscle and subcutaneous fat tissues. There was a linear relationship between the concentration of VA and cis-9,trans-11 CLA and the coefficients of determination (R (2)) varied between 0.29 and 0.87 from rumen to the different tissues. PMID- 17912567 TI - Estradiol favors the formation of eicosapentaenoic acid (20:5n-3) and n-3 docosapentaenoic acid (22:5n-3) from alpha-linolenic acid (18:3n-3) in SH-SY5Y neuroblastoma cells. AB - Whether neurosteroids regulate the synthesis of long chain polyunsaturated fatty acids in brain cells is unknown. We examined the influence of 17-beta-estradiol (E2) on the capacity of SH-SY5Y cells supplemented with alpha-linolenic acid (ALA), to produce eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA). Cells were incubated for 24 or 72 h with ALA added alone or in combination with E2 (ALA + E2). Fatty acids were analyzed by gas chromatography of ethanolamine glycerophospholipids (EtnGpl) and phosphatidylcholine (PtdCho). Incubation for 24 h with ALA alone increased EPA and DPA in EtnGpl, by 330 and 430% compared to controls (P < 0.001) and DHA by only 10% (P < 0.05). Although DHA increased by 30% (P < 0.001) in ALA + E2 treated cells, the difference between the ALA and ALA + E2 treatments were not significant after 24 h (Anova-1, Fisher's test). After 72 h, EPA, DPA and DHA further increased in EtnGpl and PtdCho of cells supplemented with ALA or ALA + E2. Incubation for 72 h with ALA + E2 specifically increased EPA (+34% in EtnGpl, P < 0.001) and DPA (+15%, P < 0.001) compared to ALA alone. Thus, SH-SY5Y cells produced membrane EPA, DPA and DHA from supplemental ALA. The formation of DHA was limited, even in the presence of E2. E2 significantly favored EPA and DPA production in cells grown for 72 h. Enhanced synthesis of ALA-elongation products in neuroblastoma cells treated with E2 supports the hypothesis that neurosteroids could modulate the metabolism of PUFA. PMID- 17912568 TI - Soy-based infant formula supplemented with DHA and ARA supports growth and increases circulating levels of these fatty acids in infants. AB - Healthy term infants (n = 244) were randomized to receive: (1) control, soy-based formula without supplementation or (2) docosahexaenoic acid-arachidonic acid (DHA + ARA), soy-based formula supplemented with at least 17 mg DHA/100 kcal (from algal oil) and 34 mg ARA/100 kcal (from fungal oil) in a double-blind, parallel group trial to evaluate safety, benefits, and growth from 14 to 120 days of age. Anthropometric measurements were taken at 14, 30, 60, 90, and 120 days of age and 24-h dietary and tolerance recall were recorded at 30, 60, 90, and 120 days of age. Adverse events were recorded throughout the study. Blood samples were drawn from subsets of 25 infants in each group. Capillary column gas chromatography was used to analyze the percentages of fatty acids in red blood cell (RBC) lipids and plasma phospholipids. Compared with the control group, percentages of fatty acids such as DHA and ARA in total RBC and plasma phospholipids were significantly higher in infants in the DHA + ARA group at 120 days of age (P < 0.001). Growth rates did not differ significantly between feeding groups at any assessed time point. Supplementation did not affect the tolerance of formula or the incidence of adverse events. Feeding healthy term infants soy-based formula supplemented with DHA and ARA from single cell oil sources at concentrations similar to human milk significantly increased circulating levels of DHA and ARA when compared with the control group. Both formulas supported normal growth and were well tolerated. PMID- 17912571 TI - Transobturator crossover readjustable sling for severe female incontinence: technique and preliminary results. AB - We present a technique that provides circumferential coaptation of the urethra as a salvage procedure for severe subset of patients. We prospectively evaluated 16 patients who had a transobturator crossover sling. Adjustable hybrid slings were used, and silicone washers were used over the anchoring columns to keep them in place and to facilitate any posterior readjustment. Mean age was 58 years and mean follow-up was 12 months. At presentation, patients had undergone at least two anti-incontinence procedures and wore a mean of six pads daily. There was a mean 93.7% overall cure in symptoms. There was one intraoperative complication (urethra perforation) that was resolved by closing the urethral wall. De novo urge incontinence developed in 2/16 patients. All patients (3/16) who had preoperative urge incontinence achieved resolution after the procedure. The transobturator crossover sling is an effective salvage procedure that may be considered in a selected subset of female patients with a nonfunctional urethra. PMID- 17912570 TI - Changes in plasma LDL and HDL composition in patients undergoing cardiac surgery. AB - Changes of lipoprotein composition have been mainly reported in conditions of sepsis. This study characterized compositional changes in LDL and HDL during the acute phase response following cardiac surgery with cardiopulmonary bypass. Twenty-one patients undergoing cardiac surgery were included in this study. Blood samples were drawn before operation and on day 2 post-surgery. In parallel to plasma lipids and antioxidant status, lipoproteins were analyzed for lipid, apolipoprotein (apo), hydroperoxide and alpha-tocopherol content. Beyond decreases in lipid concentrations and antioxidant defenses, cardiac surgery induced substantial modifications in plasma lipoproteins. ApoB decrease in LDL fraction (-46%; P < 0.0001) reflected a marked reduction in the circulating particle number. LDL cholesteryl ester content relative to apoB concentration remained unchanged post-surgery while triglyceride (+113%; P < 0.001), free cholesterol (+22%; P < 0.05) and phospholipid (+23%; P < 0.025) were raised relative to apoB indicating increased particle size. In HDL, an abrupt rise of apoSAA (P < 0.05) was observed together with a decrease of apoA1 (-22%; P < 0.005). Cholesteryl ester content in HDL fraction decreased in parallel to apoA1 concentration while triglycerides, free cholesterol and phospholipids increased relative to apoA1. In contrast to unchanged alpha-tocopherol content, hydroperoxide content was increased in LDL and HDL. By comparison to sepsis, cardiac surgery induces a comparable reduction in circulating LDL but a more limited decrease in HDL particles. Furthermore, in contrast, cardiac surgery induces an increase in polar and non-polar lipids, as well as of particle size in both LDL and HDL. PMID- 17912572 TI - Prevalence of urinary incontinence in Korean women:an epidemiologic survey. AB - The purpose of this study was to evaluate the prevalence of urinary incontinence (UI) in a Korean national community sample of female adults aged 30-79 years. During April 2003, a national Korea telephone survey using quota sampling methods was conducted. The clinically validated computer-assisted telephone interview approach was used in the survey. Of the 1,500 subjects contacted, 1,303 were successfully interviewed (response rate 86.9%). Overall, UI was reported by 40.8%, and 22.9, 3.1, and 14.9% reported pure stress, urge, and mixed UI, respectively. The prevalence of stress, urge, and mixed UI generally did not increase with age. Urge and mixed UI had a greater impact than stress UI on daily tasks (P < 0.001), social life (P < 0.001), depression or anxiety due to UI (P < 0.001), worry about UI (P < 0.001), sex life (P < 0.001), wear protection due to UI (P = 0.011), and quality of life (P < 0.001). In subjects with pure stress UI, 28.3% reported impaired quality of life compared with 43.9% and 43.8% of subjects with urge and mixed UI. Of those individuals with stress, urge, and mixed UI, 19.1, 20.0, and 25.8% had the willingness to seek medical consultation. This study is the first to examine the prevalence of UI in Korean women. The present study revealed that there was a high prevalence of UI in this population. Our descriptive research provides a valuable insight into the need for tailored education to this population about UI. PMID- 17912573 TI - Risk factors for urinary incontinence in Taiwanese women aged 60 or over. AB - To determine risk factors of urinary incontinence (UI) in Taiwanese women aged 60 or over, face-to-face interviews with 1,517 women, selected by a multistage random method, were completed. The prevalence of UI in this age group was 29.8%. Factors and their prevalence associated with UI were age [odds ratio (OR)=1.04 per year], diabetes mellitus (39.8%, p = 0.002), hypertension (39.5%, p = 0.001), abdominal gynecological surgery (41.4%, p = 0.001), hysterectomy (42.4%, p = 0.003), history of drug allergy (41.3%, p = 0.001), smoking (45.5%, p = 0.010), hormone replacement therapy (41.5%, p = 0.026), and high body mass index (OR = 1.05 per unit). Alcohol consumption and marriage did not increase the risk of UI. UI is a common and costly problem in elderly women. It diminishes the quality of life of the affected women. Of the associated factors that are preventable, modifiable, or controllable, smoking, prior hysterectomy, and obesity may have the greatest impact on the prevalence of UI. PMID- 17912574 TI - Does thoracic or lumbar spine bone architecture predict vertebral failure strength more accurately than density? AB - Trabecular bone microstructure was studied in 6 mm bone biopsies taken from the 10th thoracic and 2nd lumbar vertebra of 165 human donors and shown to not differ significantly between these sites. Microstructural parameters at the locations examined provided only marginal additional information to quantitative computed tomography in predicting experimental failure strength. INTRODUCTION: It is unknown whether trabecular microstructure differs between thoracic and lumbar vertebrae and whether it adds significant information in predicting the mechanical strength of vertebrae in combination with QCT-based bone density. METHODS: Six mm cylindrical biopsies taken at mid-vertebral level, anterior to the center of the thoracic vertebra (T) 10 and the lumbar vertebra (L) 2 were studied with micro-computed tomography (microCT) in 165 donors (age 52 to 99 years). The segment T11-L1 was examined with QCT and tested to failure using a testing machine. RESULTS: The correlation of microstructural properties was moderate between T10 and L2 (r 5 cm) and pathologic tumor stage (stage I-II and stage III-IVa). We also compared disease-free survival and the pattern of recurrence among the three groups: complete necrosis, incomplete necrosis, non-TACE groups. RESULTS: The 1-, 3-, and 5-year disease-free survival rates were 76.0%, 57.7%, and 51.3%, respectively, in the TACE group and 70.9%, 53.8%, and 46.8%, respectively, in the non-TACE group. No significant difference was observed in disease-free survival or the pattern of recurrence between the TACE and non-TACE groups. Further analysis of disease-free survival and the pattern of recurrence between subgroups according to initial tumor size and tumor stage showed no significant differences. Complete necrosis of tumor was recognized in only 33 patients of the TACE group (p = 0.001). Among the three complete necrosis, incomplete necrosis, and non-TACE groups, no significant difference was observed in disease-free survival or the pattern of recurrence. CONCLUSIONS: Preoperative TACE did not significantly improve the disease-free survival or the pattern of recurrence after curative resection of HCC. Even though this study is a retrospective analysis, preoperative TACE cannot be recommended as a routine procedure before hepatectomy for a resectable HCC. PMID- 17912588 TI - The current state of cough research: the clinician's perspective. AB - Cough is one of the most common reasons for medical consultation and it is responsible for a large human and socioeconomic burden. Current guidelines provide a useful framework for cough management and summarize current knowledge of causes and optimal testing sequences and treatments of cough. However, research is needed on the role of noninvasive airway inflammation measurement in assessing etiology; optimal treatment of postinfectious cough or cough due to gastroesophageal reflux disease; protussive treatment; causes of chronic cough in immunocompromised hosts; and characteristics and management of psychogenic cough. The effects of the use or nonuse of evidence-based guidelines should be documented. An empiric, integrative approach to management of chronic cough also needs further validation. PMID- 17912590 TI - A critical appraisal of the role of neoadjuvant chemotherapy for colorectal liver metastases: a case-controlled study. AB - BACKGROUND: The aim of this study was to analyze the outcome of patients that received neoadjuvant chemotherapy prior to resection for colorectal liver metastases (CRLM) and compare them with a matched cohort of patients that underwent resection followed by adjuvant chemotherapy. METHODS: 687 patients have undergone curative resection between January 1993 and January 2006. In this period, 84 patients received neo-adjuvant chemotherapy and 71 of this group went on to resection. A control group was chosen, matched with these patients, made up of patients who underwent resection followed by adjuvant chemotherapy. RESULTS: There was no difference in clinico-pathological features between the neoadjuvant and the control group. However patients in the control group had more-extended resections and longer hospital stays than those in the neoadjuvant group (p = 0.015). Patients in the control group had an increased incidence of early recurrences (p < 0.001). Despite this, there was no significant difference in either the cancer-specific or the disease-free survival between the two groups of patients. CONCLUSION: Neoadjuvant chemotherapy has a role in the management of patients with disease that is considered initially unresectable as a down-sizing technique. In patients with resectable disease, the test-of-time approach that neoadjuvant therapy offers is yet to be proven. PMID- 17912589 TI - Is there any prognostically different subgroup among patients with stage IIIC (any TN3M0) breast carcinoma? AB - BACKGROUND: We investigated whether there are prognostically different subgroups among patients with stage IIIC (any TN3M0) breast carcinoma. METHODS: The file records of 348 female patients operated for stage IIIC breast carcinoma were reviewed. The endpoint was disease recurrence. RESULTS: Patients with a T1, T2 or T3 tumor had significantly better disease-free survival (DFS) compared to those with a T4 tumor. In the patient group with T1,2,3N3M0 disease, the DFS was significantly better in patients with between 10 and 15 metastatic axillary lymph nodes, compared to patients with 16 or more metastatic lymph nodes (p = 0.0360) and in patients with a nodal ratio ( number of metastatic lymph nodes divided by number of removed nodes) less than or equal to 0.80, compared to patients with a nodal ratio greater than 0.80 (p = 0.0003). In the patient subgroup with between 10 and 15 metastatic lymph nodes, those with a nodal ratio greater than 0.80 had significantly worse DFS, whereas in the patient subgroup with 16 or more metastatic lymph nodes the nodal ratio had no prognostic significance. The DFS of patients with 10 to 15 positive lymph nodes and a nodal ratio of up to 0.80 was significantly better than that of both the patients with 10 to 15 positive lymph nodes and a nodal ratio greater than 0.80 (p = 0.0002), and the patients with 16 or more positive lymph nodes (p = 0.0002); survival of the latter two patient groups was similar. CONCLUSIONS: Patients with T1,2,3N3M0 disease can be divided into prognostically different subgroups according to the number of metastatic lymph nodes in the axilla and the nodal ratio; in this way, different patient subgroups may be offered different treatment strategies. PMID- 17912591 TI - A simple unforced oscillatory growth model in the chemostat. AB - In a chemostat, transient oscillations in cell number density are often experimentally observed during cell growth. The aim of this paper is to propose a simple autonomous model which is able to generate these oscillations, and to investigate it analytically. Our point of view is based on a simplification of the cell cycle in which there are two states (mature and immature) with the transfer between the two dependent on the available resources. We use the mathematical global properties of competitive differential systems to prove the existence of a limit cycle. A comparison between our model and a more complex model consisting of partial differential equations is made with the help of numerical simulations, giving qualitatively similar results. PMID- 17912592 TI - Surgical management of gastro-gastric fistula after divided laparoscopic Roux-en Y gastric bypass for morbid obesity. AB - BACKGROUND: Gastro-gastric fistula (GGF) formation is uncommon after divided laparoscopic Roux-en-Y gastric bypass (LRYGB) for morbid obesity. Optimal surgical management remains controversial. METHODS: A retrospective review was performed of a prospectively maintained database of patients undergoing LRYGB from January 2001 to October 2006. RESULTS: Of 1,763 primary procedures, 27 patients (1.5%) developed a GGF and 10 (37%) resolved with medical management, whereas 17 (63%) required surgical intervention. An additional seven patients requiring surgical intervention for GGF after RYGB were referred from another institution. Indications for surgery included weight regain, recurrent, or non healing gastrojejunal anastomotic (GJA) ulceration with persistent abdominal pain and/or hemorrhage, and/or recurrent GJA stricture. Remnant gastrectomy with GGF excision or exclusion was performed in 23 patients (96%) with an average in hospital stay of 7.5 days (range, 3-27). Morbidity in six patients (25%) was caused by pneumonia, n=2; wound infection, n=2; staple-line bleed, n=1; and subcapsular splenic hematoma, n=1. There were no mortalities. Complete resolution of symptoms and associated ulceration was seen in the majority of patients. CONCLUSION: Although uncommon, GGF formation can complicate divided LRYGB. Laparoscopic remnant gastrectomy with fistula excision or exclusion can be used to effectively manage symptomatic patients who fail to respond to conservative measures. PMID- 17912593 TI - Racial and geographic disparities in the utilization of surgical therapy for hepatocellular carcinoma. AB - The incidence of hepatocellular carcinoma (HCC) continues to increase, a trend that will likely continue because of the rising prevalence of chronic hepatitis C infection. This study sought to determine the recent patterns of utilization of surgical therapy (hepatectomy, ablation, or liver transplantation) for HCC from the Surveillance, Epidemiology, and End Results national cancer registry. Data were extracted for 16,121 patients with HCC diagnosed between 1998 and 2004. Twenty-three percent of patients underwent surgical therapy (9.5% resection, 7.8% ablation, 6% transplant); the proportion of patients treated with surgical therapy increased approximately 9% over the study period. On multivariate analysis, female sex, younger age, and smaller solitary tumors were associated with increased utilization of surgical therapy. Blacks and Hispanics were 24-27% less likely to receive surgical therapy than white individuals (P<0.001). Racial and geographic disparities persisted despite the adjustment for Health Service Area and limitation of the cohort to small localized HCC. Blacks were especially disadvantaged in the utilization of liver transplant for small HCC (OR=0.42, P<0.001). Further investigation to understand the etiology of these profound racial and geographic disparities is essential to ensure equitable provision of surgical therapies, which provide the only potentially curative treatments for HCC. PMID- 17912594 TI - Glycated type 1 collagen induces endothelial dysfunction in culture. AB - Hyperglycemia-induced protein glycation is thought to be implicated in the diabetic vasculopathy. In this study, we cultured vascular endothelial cells on native or glycated collagen matrix and compared their growth and functional characteristics. At lower plating density, the cells grew equally well on both substrata; however, at higher planting density, the cells plated on glycated collagen grew slower and reached a lower confluent density compared to that of the native collagen-based cultures. Confluent cell layers formed on glycated collagen exhibited a lower diffusion barrier function and a less response to epidermal growth factor stimulated prostacyclin production, compared to their native collagen-cultured counterparts. PMID- 17912595 TI - Amperometric glucose biosensors based on layer-by-layer assembly of chitosan and glucose oxidase on the Prussian blue-modified gold electrode. AB - A glucose biosensor based on layer-by-layer (LBL) self-assembling of chitosan and glucose oxidase (GOD) on a Prussian blue film was developed. First, Prussian blue was deposited on a cleaned gold electrode then chitosan and GOD were assembled alternately to construct a multilayer film. The resulting amperometric glucose biosensor exhibited a fast response time (within 10 s) and a linear calibration range from 6 microM to 1.6 mM with a detection limit of 3.1 microM glucose (s/n = 3). With the low operating potential, the biosensor showed little interference to the possible interferents, including ascorbic acid, acetaminophen and uric acid, indicating an excellent selectivity. PMID- 17912597 TI - Evaluation of water quality index for drinking purposes for river Netravathi, Mangalore, South India. AB - An attempt has been made to develop water quality index (WQI), using six water quality parameters Dissolved oxygen (DO), Biochemical oxygen Demand (BOD), Most Probable Number (MPN), Turbidity, Total Dissolved Solids (TDS) and pH measured at eight different stations along the river basin. Rating curves were drawn based on the tolerance limits of inland waters and health point of view. Bhargava WQI method and Harmonic Mean WQI method were used to find overall WQI along the stretch of the river basin. Five point rating scale was used to classify water quality in each of the study areas. It was found that the water quality of Netravathi varied from Excellent to Marginal range by Bhargava WQI method and Excellent to Poor range by Harmonic Mean WQI method. It was observed that the impact of human activity was severe on most of the parameters. The MPN values exceeded the tolerable limits at almost all the stations. It was observed that the main cause of deterioration in water quality was due to the lack of proper sanitation, unprotected river sites and high anthropogenic activities. PMID- 17912596 TI - Site directed mutagenesis of Drosophila flightin disrupts phosphorylation and impairs flight muscle structure and mechanics. AB - Flightin is a myosin rod binding protein that in Drosophila melanogaster is expressed exclusively in the asynchronous indirect flight muscles (IFM). Hyperphosphorylation of flightin coincides with the completion of myofibril assembly and precedes the emergence of flight competency in young adults. To investigate the role of flightin phosphorylation in vivo we generated three flightin null (fln(0)) Drosophila strains that express a mutant flightin transgene with two (Thr158, Ser 162), three (Ser139, Ser141, Ser145) or all five potential phosphorylation sites mutated to alanines. These amino acid substitutions result in lower than normal levels of flightin accumulation and transgenic strains that are unable to beat their wings. On two dimensional gels of IFM proteins, the transgenic strain with five mutant sites (fln(5STA)) is devoid of all phosphovariants, the transgenic strain with two mutant sites (fln(2TSA)) expresses only the two least acidic of the nine phosphovariants, and the transgenic strain with three mutant sites (fln(3SA)) expresses all nine phosphovariants, as the wild-type strain. These results suggest that phosphorylation of Thr158 and/or Ser162 is necessary for subsequent phosphorylation of other sites. All three transgenic strains show normal, albeit long, IFM sarcomeres in newly eclosed adults. In contrast, sarcomeres in fully mature fln(5STA) and fln(2TSA) adults show extensive breakdown while those in fln(3SA) are not as disordered. The fiber hypercontraction phenotype that characterizes fln(0) is fully evident in fln(5STA) and fln(2TSA) but partially rescued in fln(3SA). Mechanics on skinned fibers from newly eclosed flies show alterations in viscous modulus for fln(5STA) and fln(2TSA) that result in a significant reduction in oscillatory power output. Expression of fln(5STA) and fln(2TSA), but not fln(3SA), in a wild-type (fln(+)/fln(+)) background resulted in a dominant negative effect manifested as flight impairments and hypercontracted IFM fibers. Our studies indicate that Thr158 and/or Ser162 are (is) indispensable for flightin function and suggest that phosphorylation of one or both residues fulfills an essential role in IFM structural stability and mechanics. PMID- 17912598 TI - Spatial distribution of major and trace elements in the water of Swarzedzkie Lake (Poland). AB - In order to characterize the contamination of the Swarzedzkie Lake and generate elemental distribution patterns 18 elements have been determined in samples of water, taken just above the bottom sediments. Analyses were carried out using two instrumental techniques - ICP-OES and ICP-MS. Two main sources of contamination were identified - diffuse pollution from agriculture and stormwater from the town. Three separate groups of elements were distinguished using cluster analysis and two groups using principal component analysis. These three groups are characterized as geogenic elements (Ag, As, Mg and Sb), elements supplied by the agricultural catchment (Ca, Mn, Sr, Na, K and Al) and elements originated from the urban contamination (Ni, Cr, Cu, Fe, V, Ba, Zn and Pb). PCA differentiated the above group of urban elements from the rest, originated from diffuse sources, with exception of alluminium, which was situated between two distinguished groups. PMID- 17912599 TI - Gas chromatographic fingerprinting of crude oil from Idu-Ekpeye oil spillage site in Niger-delta, Nigeria. AB - Samples were collected from an oil polluted site in Niger-delta, Nigeria. Gas chromatographic analyses carried out on the samples revealed an abundance of n alkanes within the n-C8-n-C23 region. The pristane/phytane ratio of 5.70 obtained for the samples depicted a plant/terrestrial source input and a possible oxic depositional environment. The n-C17/pristane and n-C18/phytane ratios of 2.80 and 2.77, respectively, suggested that the spilled oil was only slightly weathered, as corroborated by the presence of peaks in the aromatic hydrocarbon fingerprints. The polycyclic aromatic hydrocarbon (PAH) fractions showed that the hydrocarbon fractions might have undergone combustion and/or that there was bush burning at the site prior to the oil spill incidence. This is supported by the abundance of high-molecular-weight PAHs which are pyrogenic in nature. High molecular weight PAHs are products of the combustion of petroleum or its products. The phenanthrene/anthracene ratio of 0.95, fluorathene/pyrene ratio of 2.23 and the summation operator (other three to six ringed PAHs)/ summation operator (five alkylated PAHs) ratio far greater than unity (4.10) also affirm this. On the other hand, the benzo (a) anthracene to chrysene ratio of 0.24 confirms the petrogenic origin of the spilled oil because chrysene which is highly abundant is a fossil PAH. PMID- 17912600 TI - Resistance to Grapevine leafroll associated virus-2 is conferred by post transcriptional gene silencing in transgenic Nicotiana benthamiana. AB - Grapevine leafroll-associated virus-2 (GLRaV-2) is an important component of the leafroll disease complex in grapevine. We have previously sequenced the GLRaV-2 genome and identified the coat protein (CP) gene. The objective of this study is to test the concept of pathogen-derived resistance against a closterovirus associated with grapevine leafroll disease. Because GLRaV-2 is capable of infecting Nicotiana benthamiana, we decided to test the concept on this herbaceous host. Thirty-seven T(0) transgenic N. benthamiana plants expressing the GLRaV-2 CP gene were regenerated following Agrobacterium-mediated transformation. Disease resistance was evaluated in greenhouse-grown T(1) and T(2) plants by mechanical inoculation with GLRaV-2. Although all the inoculated non-transgenic plants showed symptoms 2-4 weeks post inoculation, various numbers of transgenic plants (16-100%) in 14 of 20 T(1) lines tested were not infected. In these resistant plants, GLRaV-2 was not detectable by enzyme linked immunosorbent assay. Although virus resistance was confirmed in T(2) progenies, the percentage of resistant plants was generally lower (0-63%) than that of the corresponding T(1) lines (0-100%). Northern blot and nuclear run-off results showed that virus resistance in the transgenic plants was consistently associated with the low level of transgene RNA transcript suggesting a post-transcriptional gene silencing. The success of pathogen-derived resistance to GLRaV-2 in transgenic N. benthamiana plants represents the first step towards eventual control of the leafroll disease in grapevines using this strategy. PMID- 17912602 TI - The fluorescence yield of the trimeric fucoxanthin-chlorophyll-protein FCPa in the diatom Cyclotella meneghiniana is dependent on the amount of bound diatoxanthin. AB - The fluorescence yield of isolated fucoxanthin chlorophyll proteins, serving as light harvesting proteins in diatoms, was compared to the amount of diatoxanthin bound. Diatoxanthin was earlier shown to be involved in the xanthophyll cycle in diatoms as a functional analogue of zeaxanthin in higher plants. By growing cells under different light conditions, the amount of diatoxanthin in both the trimeric FCPa as well as the oligomeric FCPb of the diatom Cyclotella meneghiniana was increased. In the trimeric FCPa, the fluorescence yield decreased with increasing diatoxanthin content, whereas in the oligomeric FCPb fluorescence was generally lower, albeit constant. No pH dependence of fluorescence yield could be demonstrated except for artificially aggregated FCPa. Thus, diatoxanthin is able to quench fluorescence in FCPa, but the yield is also influenced by pH when the protein becomes aggregated. PMID- 17912601 TI - Human cytomegalovirus immediate-early-gene expression disrupts embryogenesis in transgenic Drosophila. AB - Intrauterine infection with human cytomegalovirus (HCMV) is the leading viral cause of birth defects involving the central nervous system. Due to the highly species specific nature of the virus, its course of natural infection cannot be studied in animal models. Here we introduce a novel transgenic Drosophila model system for studying the effects of the major viral regulatory genes, the immediate-early genes, on normal embryonic development. We show that ectopic expression of the immediate-early genes in Drosophila led to increased embryonic lethality manifested in disintegration of the embryos. Further analysis suggested that immediate-early gene expression interfered with adherens junction maintenance, leading to the disruption of embryonic epithelial integrity. Owing to the evolutionary conservation of developmental mechanisms from invertebrates to mammals, we anticipate that the studies in Drosophila will be relevant also to humans and will ultimately provide a versatile system for studying different aspects of viral-host interactions. PMID- 17912603 TI - Probing binding site of bacteriochlorophyll a and carotenoid in the reconstituted LH1 complex from Rhodospirillum rubrum S1 by Stark spectroscopy. AB - Stark spectroscopy is a powerful technique to investigate the electrostatic interactions between pigments as well as between the pigments and the proteins in photosynthetic pigment-protein complexes. In this study, Stark spectroscopy has been used to determine two nonlinear optical parameters (polarizability change Tr(Deltaalpha) and static dipole-moment change |Deltamu| upon photoexcitation) of isolated and of reconstituted LH1 complexes from the purple photosynthetic bacterium, Rhodospirillum (Rs.) rubrum. The integral LH1 complex was prepared from Rs. rubrum S1, while the reconstituted complex was assembled by addition of purified carotenoid (all-trans-spirilloxanthin) to the monomeric subunit of LH1 from Rs. rubrum S1. The reconstituted LH1 complex has its Q(y) absorption maximum at 878 nm. This is shifted to the blue by 3 nm in comparison to the isolated LH1 complex. The energy transfer efficiency from carotenoid to bacteriochlorophyll a (BChl a), which was determined by fluorescence excitation spectroscopy of the reconstituted LH1 complex, is increased to 40%, while the efficiency in the isolated LH1 complex is only 28%. Based on the differences in the values of Tr(Deltaalpha) and |Deltamu|, between these two preparations, we can calculate the change in the electric field around the BChl a molecules in the two situations to be E (Delta) approximately 3.4 x 10(5) [V/cm]. This change can explain the 3 nm wavelength shift of the Q(y) absorption band in the reconstituted LH1 complex. PMID- 17912604 TI - Nomenclature for membrane-bound light-harvesting complexes of cyanobacteria. AB - Accessory chlorophyll-binding proteins (CBP) in cyanobacteria have six transmembrane helices and about 11 conserved His residues that might participate in chlorophyll binding. In various species of cyanobacteria, the CBP proteins bind different types of chlorophylls, including chlorophylls a, b, d and divinyl chlorophyll a, b. The CBP proteins do not belong to the light-harvesting complexes (LHC) superfamily of plant and algae. The proposed new name of CBP for this class of proteins, which is a unique accessory light-harvesting superfamily in cyanobacteria, clarifies the confusion of names of prochlorophytes chlorophyll binding protein (Pcb), PSII-like light-harvesting proteins and iron-stress induced protein A (IsiA). The CBP complexes are a member of a larger family that includes the chlorophyll a-binding proteins CP43 and CP47 that function as core antennas of photosystem II. PMID- 17912605 TI - Composition and localization of bacteriochlorophyll a intermediates in the purple photosynthetic bacterium Rhodopseudomonas sp. Rits. AB - Rhodopseudomonas sp. Rits is a recently isolated new species of photosynthetic bacteria and found to accumulate a significantly high amount of bacteriochlorophyll (BChl) a intermediates possessing non-, di- and tetra hydrogenated geranylgeranyl groups at the 17-propionate as well as normal phytylated BChl a (Mizoguchi T et al. (2006) FEBS Lett 580:137-143). A phylogenetic analysis showed that this bacterium was closely related to Rhodopseudomonas palustris. The strain Rits synthesizes light-harvesting complexes 2 and 4 (LH2/4), as peripheral antennas, as well as the reaction center and light-harvesting 1 core complex (RC-LH1 core). The amounts of these complexes were dependent upon the incident light intensities, which was also a typical behavior of Rhodopseudomonas palustris. HPLC analyses of extracted pigments indicated that all four BChls a were associated with the purified photosynthetic pigment-protein, as complexes described above. The results suggested that this bacterium could use these pigments as functional molecules within the LH2/4 and RC-LH1 core. Pigment compositional analyses in several purple photosynthetic bacteria showed that such BChl a intermediates were always detected and were more widely distributed than expected. Long chains in the propionate moiety of BChl a would be one of the important factors for assembly of LH systems in purple photosynthetic bacteria. PMID- 17912606 TI - Intermediate binding of phycocyanobilin to the lyase, CpeS1, and transfer to apoprotein. AB - The phycobilin: Cysteine-84-phycobiliprotein lyase, CpeS1, catalyzes phycocyanobilin (PCB) and phycoerythrobilin attachment to nearly all cysteine-84 (consensus sequence) binding sites of phycoerythrin, phycoerythrocyanin, phycocyanin and allophycocyanin (Zhao et al. (2007) Proc Natl Acad Sci 104:14300 14305). We now show that CpeS1 can bind PCB, as assayed by Ni(2+) chelating affinity chromatography. Binding is rapid, and the chromophore is bound in an extended conformation similar to that in phycobiliproteins but only poorly fluorescent. Upon addition of apo-biliproteins, the chromophore is transferred to the latter much slower ( approximately 1 h), indicating that chromophorylated CpeS1 is an intermediate in the enzymatic reaction. In addition, imidazole is bound to PCB, as shown by mass spectroscopy of tryptic digests of the intermediate CpeS1-PCB complex. PMID- 17912607 TI - Influence of detergent concentration on aggregation and spectroscopic properties of light-harvesting complex II. AB - Aggregation of photosynthetic light-harvesting complexes strongly influences their spectroscopic properties. Fluorescence yield and excited state lifetimes of the main light-harvesting complex (LHC II) of higher plants strongly depend on its aggregation state. Detergents are commonly used to solubilize membrane proteins and/or to circumvent their aggregation in aqueous environments. Nonlinear polarization spectroscopy in the frequency domain (NLPF) was performed with LHC II over a wide concentration range of the mild detergent n-dodecyl beta D: -maltoside (beta-DM). Additionally, conventional absorption-, fluorescence- and circular dichroism-spectra were measured.The results indicate that: (i) conventional spectroscopic techniques are not well suited to investigate aggregation effects. NLPF provides a novel approach to overcome this problem: NLPF spectra display dramatic alterations upon even minor beta-DM concentration changes. (ii) Commonly used detergent concentrations (around or slightly above the critical micellar concentration) apparently do not lead to complete trimerization of LHC II. A long-wavelength species in the NLPF spectra (peaking at about 685 nm), indicative of residual aggregation, persists up to DM concentrations of 0.06%. (iii) High-resolution NLPF spectra indicate the existence of a species with a considerably shortened excited state lifetime. (iv) No indication of denaturation was found even at the highest beta-DM concentrations used. (v) A specific change in interaction between certain chlorophyll(s) b and a xanthophyll molecule, probably neoxanthin, was detected upon aggregation as well as at higher beta-DM concentrations. The results are discussed with respect to the still elusive mechanism of nonradiative dissipation of excess excitation energy in the antenna system. PMID- 17912608 TI - Self-aggregation behavior of synthetic zinc 3-hydroxymethyl-13/15-carbonyl chlorins as models of main light-harvesting components in photosynthetic green bacteria. AB - Zinc complexes of 3-hydroxymethyl-13/15-carbonyl-chlorins having a six-membered lactone as the E-ring were prepared by modifying purpurin-18 as models of bacteriochlorophyll-d, one of the chlorophyllous pigments in the main light harvesting antenna systems (chlorosomes) of green photosynthetic bacteria. The synthetic 13-carbonylated compound self-aggregated in 1%(v/v) tetrahydrofuran and hexane to give large oligomers possessing red-shifted and broadened electronic absorption bands and intense circular dichroism bands at the shifted Q ( y ) region, indicating that the supramolecular structure of the resulting self aggregate was similar to those of natural and artificial chlorosomal aggregates. The red-shift value observed here was smaller than the reported values in chlorosomal pigments having a five-membered keto-ring, which was ascribable to a weaker intermolecular hydrogen-bonding of 13-C=O with 3(1)-OH in a supramolecule of the former self-aggregate and suppression of the pi-pi interaction among the composite chlorins. On the other hand, the isomeric 15-carbonylated molecule was monomeric even in the nonpolar organic solvent, confirming the reported proposal that the linear orientation of three interactive moieties, OH, C=O and Zn, in a molecule is requisite for its chlorosomal self-aggregation. PMID- 17912609 TI - Comparison of the fluorescence kinetics of detergent-solubilized and membrane reconstituted LH2 complexes from Rps. acidophila and Rb. sphaeroides. AB - Picosecond time-resolved fluorescence spectroscopy has been used in order to compare the fluorescence kinetics of detergent-solubilized and membrane reconstituted light-harvesting 2 (LH2) complexes from the purple bacteria Rhodopseudomonas (Rps.) acidophila and Rhodobacter (Rb.) sphaeroides. LH2 complexes were reconstituted in phospholipid model membranes at different lipid:protein-ratios and all samples were studied exciting with a wide range of excitation densities. While the detergent-solubilized LH2 complexes from Rps. acidophila showed monoexponential decay kinetics (tau(f )= 980 ps) for excitation densities of up to 3.10(13) photons/(pulse.cm(2)), the membrane-reconstituted LH2 complexes showed multiexponential kinetics even at low excitation densities and high lipid:protein-ratios. The latter finding indicates an efficient clustering of LH2 complexes in the phospholipid membranes. Similar results were obtained for the LH2 complexes from Rb. sphaeroides. PMID- 17912610 TI - Association of small CAB-like proteins (SCPs) of Synechocystis sp. PCC 6803 with Photosystem II. AB - The cyanobacterial small CAB-like proteins (SCPs) are one-helix proteins with compelling similarity to the first and third transmembrane helix of proteins belonging to the CAB family of light-harvesting complex proteins in plants. The SCP proteins are transiently expressed at high light intensity and other stress conditions but their exact function remains largely unknown. Recently we showed association of ScpD with light-stressed, monomeric Photosystem II in Synechocystis sp. PCC 6803 (Yao et al. J Biol Chem 282:267-276, 2007). Here we show that ScpB associates with Photosystem II at normal growth conditions. Moreover, upon introduction of a construct into Synechocystis so that ScpB is expressed continuously under normal growth conditions, ScpE was detected under non-stressed conditions as well, and was copurified with tagged ScpB and Photosystem II. We also report on a one-helix protein, Slr1544, that is somewhat similar to the SCPs and whose gene is cotranscribed with that of ScpD; Slr1544 is another member of the extended light-harvesting-like (Lil) protein family, and we propose to name it LilA. PMID- 17912612 TI - Hypermetabolism in Gaucher disease type I is not associated with altered thyroid hormone levels. AB - Type I Gaucher disease (OMIM 231000) is an inherited storage disorder in which deficiency of the enzyme glucocerebrosidase (EC 32145) leads to accumulation of glucocerebroside in lysosomes of macrophages. These storage cells are present in liver, spleen and bone marrow resulting in hepatosplenomegaly, cytopenia and bone complications. Metabolic abnormalities in Gaucher patients include hypermetabolism, possibly caused by elevated levels of pro-inflammatory cytokines. Nonthyroidal illness (NTI) is a combination of changes in circulating thyroid hormone levels (decreased T(3), elevated rT(3), normal or mildly depressed TSH) present in different illnesses and might be an adaptation to protect the organism from harmful catabolic effects of hypermetabolism. The hypermetabolism and the elevated cytokine levels in Gaucher disease led us to hypothesize that the alterations in thyroid hormone levels as seen in NTI might also occur in Gaucher patients. We studied thyroid hormone levels before and during treatment in 22 adult type I Gaucher patients and resting energy expenditure (REE) and correlations with thyroid hormone levels in 12 patients. Baseline thyroid hormone levels were normal in the majority (17) of patients. No cases of nonthyroidal illness were detected. Baseline REE (kcal/kg per 24 h) was not correlated with circulating levels of T(3), rT(3) or fT(4). Treatment of Gaucher disease with enzyme replacement therapy for several years resulted in a decrease in circulating fT(4) levels. After several months of treatment most patients showed a decrease in REE. There was no correlation between the changes in REE and changes in fT(4) and T(3). PMID- 17912611 TI - Evolution of early eukaryotic cells: genomes, proteomes, and compartments. AB - Eukaryotes arose from an endosymbiotic association of an alpha-proteobacterium like organism (the ancestor of mitochondria) with a host cell (lacking mitochondria or plastids). Plants arose by the addition of a cyanobacterium-like endosymbiont (the ancestor of plastids) to the two-member association. Each member of the association brought a unique internal environment and a unique genome. Analyses of recently acquired genomic sequences with newly developed algorithms have revealed (a) that the number of endosymbiont genes that remain in eukaryotic cells-principally in the nucleus-is surprisingly large, (b) that protein products of a large number of genes (or their descendents) that entered the association in the genome of the host are now directed to an organelle derived from an endosymbiont, and (c) that protein products of genes traceable to endosymbiont genomes are directed to the nucleo-cytoplasmic compartment. Consideration of these remarkable findings has led to the present suggestion that contemporary eukaryotic cells evolved through continual chance relocation and testing of genes as well as combinations of gene products and biochemical processes in each unique cell compartment derived from a member of the eukaryotic association. Most of these events occurred during about 300 million years, or so, before contemporary forms of eukaryotic cells appear in the fossil record; they continue today. PMID- 17912613 TI - Noninvasive measurement of phenylalanine by iontophoretic extraction in patients with phenylketonuria. AB - Phenylketonuria is an autosomal recessive disorder characterized by elevated concentrations of phenylalanine. Elevated phenylalanine concentrations can impair intellectual functions and the disease is treated with a lifelong diet and frequent monitoring of plasma phenylalanine concentrations. Previous in vitro studies have demonstrated the feasibility of iontophoretically enhanced transdermal transport of phenylalanine. Here we evaluate the feasibility of transdermal iontophoretic extraction of phenylalanine in vivo. Phenylalanine was iontophoretically extracted from the skin of healthy volunteers and of patients with phenylketonuria for up to 6 h and concentrations were compared with those measured in plasma. The amount of phenylalanine iontophoretically extracted from the skin declined over time, suggesting contribution of phenylalanine from the skin in the initial extraction. Phenylalanine iontophoretically extracted from skin correlated with plasma phenylalanine levels at plasma levels above 300 micromol/L. This correlation supports the feasibility of iontophoretic phenylalanine extraction for monitoring phenylketonuria. PMID- 17912615 TI - Redescriptions of Monopylidium exiguum (Dujardin, 1845) and M. albani (Mettrick, 1958) n. comb. (Cestoda: Dilepididae) from European passerine birds. AB - Two species of Monopylidium Fuhrmann, 1899 are redescribed on the basis of specimens from their type-hosts: M. exiguum (Dujardin, 1845) from Troglodytes troglodytes L. (Passeriformes: Troglodytidae) on the Curonian Spit, Kaliningradskaya Oblast', Russia, and M. albani (Mettrick, 1958) n. comb. (originally Paricterotaenia albani Mettrick, 1958) from Sturnus vulgaris L. (Passeriformes: Sturnidae) in the Canton of Jura, Switzerland. In contrast to the previously proposed synonymy of these two species (Spasskaya & Spasskii, 1977), they are recognised as distinct and M. albani is revalidated. PMID- 17912614 TI - Marker states and a health state prompt provide modest improvements in the reliability and validity of the standard gamble and rating scale in prostate cancer patients. AB - PURPOSE: To evaluate the combined effect of marker states and a prompt on the reliability and validity of direct utility assessment. METHODS: In a structured interview, 141 prostate cancer (PC) patients provided rating scale (RS) and standard gamble (SG) utilities for their "own health" (RS-/SG-). Following self completion of comparison instruments (two generic utility and one disease specific psychometric), they answered a checklist of PC-related items (a prompt to consider self-health) and provided utilities for self-health and mild and severe PC marker states (RS+/SG+). The interview was repeated 5 weeks later, but without comparison instruments. Using Bayesian modeling, we computed and compared correlation coefficients to assess RS and SG test-retest reliability and validity and the effects of the prompt and marker states. RESULTS: RS and SG had acceptable test-retest reliability (intraclass correlation coefficients = 0.57 0.63). The prompt and marker states decreased the reliability of the RS by 0.01 (from 0.58 to 0.57) but increased the reliability of the SG by 0.05 (from 0.58 to 0.63). The probability that the reliability of the SG+ was greater than that of the SG- was very high (0.96). Correlations with comparison instruments were higher by 0.01-0.06 for RS+ vs RS-, and higher by 0.03-0.06 for SG+ vs SG-. The probabilities that the prompt and marker states improved validity ranged from 0.55 to 0.74 (RS), and from 0.61 to 0.70 (SG). CONCLUSIONS: A self-health description prompt and marker states modestly improved the reliability and validity of direct utility elicitation. PMID- 17912616 TI - Naricolax hoi n. sp. (Cyclopoida: Bomolochidae) from Arius maculatus (Siluriformes: Ariidae) off Taiwan and a redescription of N. chrysophryenus (Roubal, Armitage & Rohde, 1983) from a new host, Seriola lalandi (Perciformes: Carangidae), in Australian waters. AB - We propose that Naricolax stocki (Roubal, 1981) (Cyclopoida: Bomolochidae) of Ho & Lin (2005), reported from the spotted catfish Arius maculatus (Thunburg) off Taiwan, represents a new species, N. hoi n. sp. N. hoi can be distinguished from six known congeners by the shape of the rostral area, the maxillary armature and the structural details of legs 3 and 4. N. chrysophryenus (Roubal, Armitage & Rohde, 1983) is redescribed on the basis of recently collected material from wild and farmed yellowtail kingfish Seriola lalandi Valenciennes in southern and eastern Australian waters, providing the first record of Naricolax Ho, Do & Kasahara, 1983 from a carangid host. A key to the species of Naricolax is provided. PMID- 17912617 TI - Allomermis solenopsi n. sp. (Nematoda: Mermithidae) parasitising the fire ant Solenopsis invicta Buren (Hymenoptera: Formicidae) in Argentina. AB - Allomermis solenopsi n. sp. (Mermithidae: Nematoda) is described from the fire ant Solenopsis invicta Buren (Hymenoptera: Formicidae) in Argentina. Diagnostic characters of the new species include stiff and erect processes on the surface of the mature egg, small female amphids, extension of the latero-medial rows of male genital papillae beyond the middle rows, an obliquely truncate spicule tip and a ventrally swollen male terminus. This is the first record of Allomermis Steiner, 1924 from South America and the first host record for members of this genus. Previous records of mermithids from Solenopsis spp. are summarised. The placement in Allomermis was confirmed by molecular analyses based on nuclear 18S ribosomal DNA sequences, the first such molecular framework for the Mermithidae. The possible life-cycle of the parasite is discussed, with the aim of using A. solenopsi as a biological control agent for fire ants in the United States. PMID- 17912618 TI - The morphology and systematic status of Khawia rossittensis (Szidat, 1937) and K. parva (Zmeev, 1936) (Cestoda: Caryophyllidea), parasites of cyprinid fishes. AB - The morphology of the two little-known fish cestodes of the genus Khawia Hsu, 1935, K. rossittensis (Szidat, 1937) and K. parva (Zmeev, 1936) from cyprinid fishes, were studied on the basis of newly collected specimens from goldfish Carassius auratus auratus (L.) and museum specimens, respectively. This paper provides the first detailed species diagnosis for K. rossittensis from Slovakia, which is compared with specimens from different geographical regions and K. parva, a somewhat similar Far Eastern species from the same host. The two species differ in scolex morphology, anterior extent of the vitelline follicles, shape of the ovary and size of the eggs. Based on these differences, K. rossittensis and K. parva are considered to be separate taxa. K. parva, listed among the "species incertae sedis" by Mackiewicz (1972) and even within Caryophyllaeus Gmelin, 1790 by Schmidt (1986), should be considered a valid species of Khawia. The results support the previous conclusions of Kulakovskaya (1961), Dubinina (1971) and Protasova et al. (1990). PMID- 17912619 TI - A description of the parthenogenetic metacercaria and cercaria of Cercaria falsicingulae I larva nov. (Digenea: Gymnophallidae) from the snails Falsicingula spp. (Gastropoda), with speculation on an unusual life-cycle. AB - Previously unknown gymnophallid parthenogenetic metacercariae (PM), referred to as Cercaria falsicingulae I larva nov., were found in the extrapallial cavities of the snails Falsicingula mundana (Yokoyama) and F. athera (Bartsch) on coasts of Sakhalin and the Kuril Islands. Unlike all other known PM, rather than producing metacercariae infective to the definitive host, their furcocercariae emerge into the environment. The developing cercariae and metacercariae of C. falsicingulae I are described and compared with other gymnophallid larvae from littoral molluscs in the region. Experimental evidence and analysis of metacercarial group composition in naturally infected molluscs indicate that some cercariae leave their molluscan hosts and penetrate other specimens of Falsicingula in which they develop into new cercariae-producing PM. Metacercariae with developing hermaphroditic reproductive organs were never observed in naturally infected molluscs. A probable life-cycle for C. falsicingulae I is presented in which cercarial/metacercarial production is switched seasonally in order to enable the infection of the definitive hosts by PM. Importantly, the 'cercaria - PM - cercaria' component of the life-cycle appears to be somewhat autonomous. PMID- 17912620 TI - Morphology, development and probable systematic position of Cercariaeum crassum Wesenberg-Lund, 1934 (Digenea), a parasite of Pisidium amnicum in eastern Finland. AB - Cercariaeum crassum Wesenberg-Lund, 1934 is redescribed at the cercariaeum stage and the daughter-rediae and cercaria are also described on the basis of new material from Pisidium amnicum collected in the Liikasepuro River (eastern Finland). The species is allocated to the family Allocreadiidae, although its generic affiliation remains unknown. The probable life-cycle (based on the developmental stages observed in daughter-redia) appears to eliminate the cercarial stage and, instead, a cercariaeum (a type of cercaria without a tail) may develop directly from germ balls or, rarely, through the stages of an ophthalmoxiphidiocercaria that transforms into a young caudate cercariaeum. Their morphology and development are shown to be consistent with the family Allocreadiidae. The probable lack of a second intermediate host in the life-cycle is discussed. PMID- 17912622 TI - Repeated low-dose of erythropoietin is associated with improved left ventricular function in rat acute myocardial infarction model. AB - OBJECTIVE: To evaluate the potential protective affects of Epo on left ventricular (LV) function and remodeling after acute myocardial infarction (MI). METHODS: Epo was injected into the peritoneum of male Wistar rats (250 g) during 6 weeks post induction of MI. Rats were divided into five groups: MI treated with single high dose (MT1, 5,000 U/kg, n=10), single high dose (5,000 U/kg) and repeated high doses (MTHi, 1,000 U/kg twice a week; n=8), or single high dose (5,000 U/kg) and repeated low doses (MTLo, 750 U/kg once a week, n=10), MI non treated (MNT, n=10), sham (S, n=5). Echocardiography was performed 3.6+/-1.5 days and 43.7+/-2.3 days post MI. Collagen deposition and infarct size were measured on histological sections using computerized image analysis. Apoptosis was assessed by ApopTag staining. RESULTS: Baseline fractional shortening (FS) was similar between groups. Six weeks after MI the FS of MTLo (26.9%) was significantly higher compared to MNT (17.8%), MT1 (19.5%) and MTH (22.3%) (p=0.01). However, remodeling indices (end diastolic and end systolic areas, LV circumference) did not improve in the Epo groups, and even worsened in the MTHi group. There was significantly less collagen staining in non-infarct areas in MT1 and MTHi groups compared to MNT and MTLo (0.38+/-0.3%, 0.49+/-0.34%, vs 0.89+/ 0.41%, 0.95+/-0.33%, respectively, p<0.001). The number of ApopTag positive nucleus was significantly higher in the MNT group compared to the MT1, MTHi, MTLo groups (14.4+/-8, 7.6+/-4, 5.8+/-7, 4.8+/-5, respectively, p=0.01 for trend). CONCLUSION: Repeated low doses of Epo after MI improved LV function, but the role of Epo on remodeling is not clear. It did not reduce left ventricular indices, but reduces fibrosis and apoptosis. High Epo doses reduced LV function and aggravated remodeling. PMID- 17912623 TI - Lost in summation: depression among African American female caregivers and noncaregivers. AB - Ethnic differences in mental health have been established using large between group research designs. Across ethnicity, studies have found that caregivers are at increased risk for depression, but little is known about within-group variability in depressive symptomatology. African American caregivers and noncaregivers were compared on different factors of depressive symptoms as measured by the Center for Epidemiologic Studies Depression subscales. Caregivers reported significantly less positive affect than noncaregivers. Rates were similar for negative affect, somatic complaints, and interpersonal relations. Depression may present itself in different ways among African Americans in the caregiving context, and results suggest information may be lost when global measures of depression are used. PMID- 17912624 TI - Testing structural models of DSM-IV symptoms of common forms of child and adolescent psychopathology. AB - Confirmatory factor analyses were conducted of Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) symptoms of common mental disorders derived from structured interviews of a representative sample of 4,049 twin children and adolescents and their adult caretakers. A dimensional model based on the assignment of symptoms to syndromes in DSM-IV fit better than alternative models, but some dimensions were highly correlated. Modest sex and age differences in factor loadings and correlations were found that suggest that the dimensions of psychopathology are stable across sex and age, but slightly more differentiated at older ages and in males. The dimensions of symptoms were found to be hierarchically organized within higher-order "externalizing" and "internalizing" dimensions, which accounted for much of their variance. Major depression and generalized anxiety disorder were substantially correlated with both the "externalizing" dimension and the "internalizing" dimension, however, suggesting the need to reconceptualize the nature of these higher-order dimensions. PMID- 17912625 TI - The neuroprotective potential of phase II enzyme inducer on motor neuron survival in traumatic spinal cord injury in vitro. AB - (1) Phase II enzyme inducer is a kind of compound which can promote the expression of antioxidative enzymes through nuclear factor erythroid 2-related factor 2 (Nrf2) activation. Recently, it has been reported that these compounds show neuroprotective effect via combating oxidative stress. The purpose of this study is to determine whether phase II enzyme inducers have neuroprotective effects on traumatic spinal cord injury. (2) An organotypic spinal cord culture system was used, Phase II enzyme inducers were added to culture medium for 1 week, motor neurons were counted by SMI-32 staining, glutamate, Nrf2, and Heme oxygenase-1(HO-1) mRNA were tested. (3) This study showed motor neuron loss within 1 week in culture. After 1 week in culture, the system was stable. Moreover, Glutamate was increased when in culture 48 h and decreased after 1 week in culture. There was no significant change between 1 and 4 weeks in culture. Necrotic motor neuron and damaged mitochondrial were observed in culture 48 h. Furthermore, phase II enzyme inducers: tert-butyhydroquinone (t-BHQ), 3H-1,2 dithiole-3-thione (D3T), and 5,6-dihydrocyclopenta-1,2-dithiole-3-thione (CPDT) were shown to promote motor neuron survival after dissection, it was due to increasing Nrf2 and HO-1 mRNA expression and protecting mitochondrial not due to decreasing glutamate level. (4) The loss of motor neuron due to dissection can mimic severe traumatic spinal cord injury. These results demonstrate that glutamate excitotoxicity and the damage of mitochondrial is possibly involve in motor neuron death after traumatic spinal cord injury and phase II enzyme inducers show neuroprotective potential on motor neuron survival in traumatic spinal cord injury in vitro. PMID- 17912626 TI - Construction of SH-EP1-alpha4beta2-hAPP695 cell line and effects of nicotinic agonists on beta-amyloid in the cells. AB - (1) Nicotinic acetylcholine receptors in central nervous system are thought to be new targets for Alzheimer's disease. However, the most involved nicotinic receptor subtype in Alzheimer's disease is unclear. alpha4beta2 receptor is the most widely spread subtype in brain, involving in several important aspects of cognitive and other functions. We constructed cell line by transfecting human amyloid precursor protein (695) gene into SH-EP1 cells which have been transfected with human nicotinic receptor alpha4 subunit and beta2 subunit gene, to observe effects of alpha4beta2 receptors activation on beta-amyloid, expecting to provide a new cell line for drug screening and research purpose. (2) Liposome transfection was used to express human amyloid precursor protein (695) gene in SH EP1-alpha4beta2 cells. Function of the transfected alpha4beta2 receptors was tested by patch clamp. Effects of nicotine and epibatidine (selective alpha4beta2 nicotinic receptor agonist) on beta-amyloid were detected by Western blot and ELISA. Effects of nicotine and epibatidine on amyloid precursor protein (695) mRNA level were measured using real-time PCR. (3) Human amyloid precursor protein (695) gene was stably expressed in SH-EP1-alpha4beta2 cells; Nicotine (1 muM) and epibatidine (0.1 muM) decreased intracellular and secreted beta-amyloid in the cells; and activation of alpha4beta2 receptors did not affect amyloid precursor protein (695) mRNA level. (4) These results suggest that the constructed cell line, expressing both amyloid precursor protein (695) gene and human nicotinic receptor alpha4 subunit and beta2 subunit gene, might be useful for screening specific nicotinic receptor agonists against Alzheimer's disease. Alteration of Abeta level induced by activation of alpha4beta2 nAChR in our study might occur at a post-translational level. PMID- 17912628 TI - Neuroscience in China. Introduction. AB - The edition of a special issue entitled "Neuroscience in China" provides a unique opportunity to introduce neuroscience research undertakings in China today. Among the biological sciences at large, neuroscience is one of the most advanced fields in China. To take two examples, more than 70% of the articles published in the Chinese Journal of Physiology were in the category of neuroscience; among the members of the Chinese Academy of Sciences (CAS), scientists in the field of neuroscience outnumber scientists in other disciplines of basic medical sciences. This assay tries to analyze the historical and contemporary background underlying the current status. PMID- 17912627 TI - Melatonin regulates the viability and differentiation of rat midbrain neural stem cells. AB - (1) Neurogenesis driven by neural stem cells (NSCs) is regulated by physiological and pathological factors. Melatonin (MT) has profound neurotrophic and neuroprotective effects. Hence, we studied the role of MT in regulating the viability and differentiation of NSCs derived from rat ventral midbrain. (2) NSCs were isolated from the rat ventral midbrain. The viability of NSCs was determined by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-ulfophenyl)-2H tetrazolium assay. The differentiation of NSCs was examined by analyzing the expression of the neural markers, MT receptors, brain derived neurotropic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) with semi quantitative RT-PCR, immunofluorescence cytochemistry, and Western blot. (3) Our results showed that MT could promote the viability of NSCs. In addition, MT could significantly elevate the mRNA and protein levels of tyroxine hydroxylase (TH), a marker of dopaminergic neurons, and decrease the expression of the astrocytes maker glial fibrillary acidic protein (GFAP). MT also increased the production of BDNF and GDNF in the cultured NSCs. Meanwhile, we first found that two subtypes of MT receptors, MT1 and MT2, were expressed in the ventral midbrain NSCs. (4) These results demonstrated that MT could induce NSCs to differentiate into dopaminergic neurons and decrease astrocyte production. These findings also suggest that MT could offer a beneficial tool in guiding directional differentiation of NSCs. PMID- 17912629 TI - Automated quantitative analysis of estrogen receptor expression in breast carcinoma does not differ from expert pathologist scoring: a tissue microarray study of 3,484 cases. AB - BACKGROUND: Estrogen receptor (ER) expression is routinely assessed by immunohistochemistry (IHC) in breast carcinoma. Our study compares visual scoring of ER in invasive breast cancer by histopathologists to quantitation of staining using a fully automated system. MATERIALS AND METHODS: A tissue microarray was constructed from 4,049 cases (3,484 included in analysis) of invasive breast carcinoma linked to treatment and outcome information. Slides were scored independently by two pathologists and scores were dichotomised, with ER positivity recognized at a cut-off of >1% positive nuclei. The slides were scanned and analyzed with an Ariol automated system. RESULTS: Using data dichotomised as ER positive or negative, both visual and automated scores were highly consistent: there was excellent concordance between two pathologists (kappa = 0.918 (95%CI: 0.903-0.932)) and between two Ariol machines (kappa = 0.913 (95%CI: 0.897-0.928)). The prognostic significance of ER positivity was similar whether determined by pathologist or automated scoring for both the entire patient cohort and subsets of patients treated with tamoxifen alone or receiving no systemic adjuvant therapy. The optimal cut point for the automated scores using breast cancer disease-specific survival as an endpoint was >0.4% positive nuclei. The concordance between dextran-coated charcoal ER biochemical assay data and automated scores (kappa = 0.728 (95%CI: 0.69-0.75); 0.74 (95%CI: 0.71-0.77)) was similar to the concordance between biochemical assay and pathologist scores (kappa = 0.72 (95%CI: 0.70-0.75; 0.70 (95%CI: 0.67-0.72)). CONCLUSION: Fully automated quantitation of ER immunostaining yields results that do not differ from human scoring against both biochemical assay and patient outcome gold standards. PMID- 17912630 TI - Distribution and significance of caveolin 2 expression in normal breast and invasive breast cancer: an immunofluorescence and immunohistochemical analysis. AB - BACKGROUND: The aims of this study were to define the distribution of caveolin 2 (CAV2) in frozen and formalin fixed, paraffin embedded (FFPE) normal breast samples and the significance of CAV2 expression in breast cancer. METHODS: Caveolin 2 distribution in frozen and paraffin-embedded whole tissue sections of normal breast was evaluated using immunohistochemistry and immunofluorescence, in conjunction with antibodies to define luminal epithelial cells (oestrogen receptor and cytokeratin 8/18) and myoepithelial/ basal cells (cytokeratins 14 and 5/6, p63 and smooth muscle actin). CAV2 expression was also immunohistochemically analysed in two independent cohorts of invasive breast carcinomas (n = 245 and n = 418). RESULTS: In normal breast, CAV2 was expressed in myoepithelial cells, endothelial cells, fibroblasts and adipocytes. Luminal epithelial cells showed no or only negligible staining. CAV2 expression was observed in 9.6% of all breast cancers and was strongly correlated with high histological grade, lack of oestrogen receptor, progesterone receptor and cyclin D1 expression, and positivity for epidermal growth factor receptor, basal markers, p53 expression, and high proliferation index. Furthermore, CAV2 expression was significantly associated with basal-like immunophenotype and proved to be a prognostic factor for breast cancer-specific survival on univariate analysis. CONCLUSION: Our results demonstrate that CAV2 is preferentially expressed in basal-like cancers and is associated with poor prognosis. Further in vitro studies are required to determine whether CAV2 has oncogenic properties or is only a surrogate marker of basal-like carcinomas. PMID- 17912632 TI - The early days of letrozole. PMID- 17912633 TI - The discovery and mechanism of action of letrozole. AB - Because estrogen contributes to the promotion and progression of breast cancer, a greater understanding of the role of estrogen in breast cancer has led to therapeutic strategies targeting estrogen synthesis, the estrogen receptor, and intracellular signaling pathways. The enzyme aromatase catalyses the final step in estrogen biosynthesis and was identified as an attractive target for selective inhibition. Modern third-generation aromatase inhibitors (AIs) effectively block the production of estrogen without exerting effects on other steroidogenic pathways. The discovery of letrozole (Femara) achieved the goal of discovering a highly potent and totally selective AI. Letrozole has greater potency than other AIs, including anastrozole, exemestane, formestane, and aminoglutethimide. Moreover, letrozole produces near complete inhibition of aromatase in peripheral tissues and is associated with greater suppression of estrogen than is achieved with other AIs. The potent anti-tumor effects of letrozole were demonstrated in several animal models. Studies with MCF-7Ca xenografts successfully predicted that letrozole would be clinically superior to the previous gold standard tamoxifen and also indicated that it may be more effective than other AIs. An extensive program of randomized clinical trials has demonstrated the clinical benefits of letrozole across the spectrum of hormone-responsive breast cancer in postmenopausal women. PMID- 17912634 TI - Letrozole in the neoadjuvant setting: the P024 trial. AB - Neoadjuvant chemotherapy trials have consistently reported lower response rates in hormone receptor-positive (HR+) breast cancer when compared with HR- cases. Preoperative endocrine therapy has therefore become a logical alternative and has gained considerable momentum from the finding that aromatase inhibitors (AIs) are more effective than tamoxifen for HR+ breast cancer in both the neoadjuvant and adjuvant settings. The most convincing neoadjuvant trial to demonstrate the superiority of an AI versus tamoxifen was the P024 study, a large multinational double-blind trial in postmenopausal women with HR+ breast cancer ineligible for breast-conserving surgery. The overall response rate (ORR) was 55% for letrozole and 36% for tamoxifen (P<0.001). Significantly more letrozole-treated patients underwent breast-conserving surgery (45 vs. 35%, respectively; P=0.022). In addition, ORR was significantly higher with letrozole than tamoxifen in the human epidermal growth factor receptor HER1/HER2+ subgroup (P=0.0004). The clinical efficacy of letrozole in HER2+ breast cancer was confirmed by fluorescent in situ hybridization analysis and was found to be comparable to that of HER2- cases (ORR 71% in both subsets). Biomarker studies confirmed the superiority of letrozole in centrally assessed estrogen receptor-positive (ER+) tumors and found a strong relationship with the degree of ER positivity for both agents. Interestingly, letrozole was effective even in marginally ER+ tumors and, unlike tamoxifen, consistently reduced the expression from estrogen-regulated genes (progesterone receptor and trefoil factor 1). Furthermore, when analyzed by Ki67 immunohistochemistry, letrozole was significantly more effective than tamoxifen in reducing tumor proliferation (P=0.0009). Thus, neoadjuvant letrozole is safe and superior to tamoxifen in the treatment of postmenopausal women with HR+ locally advanced breast cancer. PMID- 17912635 TI - Letrozole in the extended adjuvant setting: MA.17. AB - Relapse after completing adjuvant tamoxifen therapy is a persistent threat for women with hormone-responsive breast cancer. Third-generation aromatase inhibitors, such as letrozole, provide a new option for extended adjuvant hormonal therapy after 5 years of tamoxifen. MA.17 was conducted to determine whether letrozole improves outcome after discontinuation of tamoxifen. Postmenopausal women with hormone receptor-positive breast cancer (N=5,187) were randomized to letrozole 2.5 mg or placebo once daily for 5 years. At a median follow-up of 30 months, letrozole significantly improved disease-free survival (DFS; P<0.001), the primary end point, compared with placebo (hazard ratio [HR] for recurrence or contralateral breast cancer 0.58; 95% confidence interval [CI] 0.45, 0.76] P<0.001). Furthermore, letrozole significantly improved distant DFS (HR=0.60; 95% CI 0.43, 0.84; P=0.002) and, in women with node-positive tumors, overall survival (HR=0.61; 95% CI 0.38, 0.98; P=0.04). Clinical benefits, including an overall survival advantage, were also seen in women who crossed over from placebo to letrozole after unblinding, indicating that tumors remain sensitive to hormone therapy despite a prolonged period since discontinuation of tamoxifen. The efficacy and safety of letrozole therapy beyond 5 years is being assessed in a re-randomization study, following the emergence of new data suggesting that clinical benefit correlates with the duration of letrozole. MA.17 showed that letrozole is extremely well-tolerated relative to placebo. Letrozole should be considered for all women completing tamoxifen; new results from the post-unblinding analysis suggest that letrozole treatment should also be considered for all disease-free women for periods up to 5 years following completion of adjuvant tamoxifen. PMID- 17912636 TI - Letrozole as upfront endocrine therapy for postmenopausal women with hormone sensitive breast cancer: BIG 1-98. AB - The BIG 1-98 trial is a large, randomized, independently conducted clinical trial designed to compare the efficacy of upfront letrozole versus tamoxifen monotherapy and to compare sequential or up-front use of letrozole and/or tamoxifen as an early adjuvant therapy for patients with early breast cancer. We report on the results from the primary core analysis of the BIG 1-98 trial of 8,010 patients, which compares monotherapy with letrozole versus tamoxifen. This pre-planned core analysis allowed the use of patient data from the monotherapy arms of letrozole and tamoxifen and from the sequential arms prior to the drug switch point. Patients randomized to letrozole had a 19% improved disease-free survival (hazard ratio [HR]=0.81; P=0.003), due especially to reduced distant metastases (HR=0.73; P=0.001). A 14% risk reduction of fatal events in favor of letrozole was also observed (P=NS). The results from the monotherapy arms alone confirmed the findings from the primary core analysis. Based on the results from this trial, the aromatase inhibitor letrozole (Femara) is currently recommended as a part of standard adjuvant therapy for postmenopausal women with endocrine responsive breast cancer and has recently been approved in the early adjuvant setting in both Europe and the United States. A subsequent analysis after additional follow-up will address the question of monotherapy versus sequential therapy. PMID- 17912637 TI - A decade of letrozole: FACE. AB - Third-generation nonsteroidal aromatase inhibitors (AIs), letrozole and anastrozole, are superior to tamoxifen as initial therapy for early breast cancer but have not been directly compared in a head-to-head adjuvant trial. Cumulative evidence suggests that AIs are not equivalent in terms of potency of estrogen suppression and that there may be differences in clinical efficacy. Thus, with no data from head-to-head comparisons of the AIs as adjuvant therapy yet available, the question of whether there are efficacy differences between the AIs remains. To help answer this question, the Femara versus Anastrozole Clinical Evaluation (FACE) is a phase IIIb open-label, randomized, multicenter trial designed to test whether letrozole or anastrozole has superior efficacy as adjuvant treatment of postmenopausal women with hormone receptor (HR)- and lymph node-positive breast cancer. Eligible patients (target accrual, N=4,000) are randomized to receive either letrozole 2.5 mg or anastrozole 1 mg daily for up to 5 years. The primary objective is to compare disease-free survival at 5 years. Secondary end points include safety, overall survival, time to distant metastases, and time to contralateral breast cancer. The FACE trial will determine whether or not letrozole offers a greater clinical benefit to postmenopausal women with HR+ early breast cancer at increased risk of early recurrence compared with anastrozole. PMID- 17912638 TI - Safety of aromatase inhibitors in the adjuvant setting. AB - The third-generation aromatase inhibitors (AIs) letrozole, anastrozole, and exemestane are replacing tamoxifen as adjuvant therapy in most postmenopausal women with early breast cancer. Although AIs have demonstrated superior efficacy and better overall safety compared with tamoxifen in randomized controlled trials, they may not provide the cardioprotective effects of tamoxifen, and bone loss may be a concern with their long-term adjuvant use. Patients require regular bone mineral density monitoring, and prophylactic bisphosphonates are being evaluated to determine whether they may protect long-term bone health. AIs decrease the risks of thromboembolic and cerebrovascular events compared with tamoxifen, and the overall rate of cardiovascular events in patients treated with AIs is within the range seen in age-matched, non-breast-cancer populations. AIs are also associated with a lower incidence of endometrial cancer and fewer vaginal bleeding/discharge events than tamoxifen. Compared with tamoxifen, the incidence of hot flashes is lower with anastrozole and letrozole but may be higher with exemestane. Generally, adverse events with AIs are predictable and manageable, whereas tamoxifen may be associated with life-threatening events in a minority of patients. Overall, the benefits of AIs over tamoxifen are achieved without compromising overall quality of life. PMID- 17912641 TI - Absence of Bax switched MG132-induced apoptosis to non-apoptotic cell death that could be suppressed by transcriptional or translational inhibition. AB - Targeting to the ubiquitin proteasome degradation pathway has become a promising approach for treating cancer. Previous studies showed that inhibition of the proteasome can induce apoptosis in various cancer cells. However, whether and how the inhibition of the proteasome induces other forms of cell death is not quite known. We previously showed that proteasome inhibitors including MG132 and Bortezomib could induce apoptosis in a Bax- and caspase-dependent way. In the present study, we found that in the absence of Bax and caspase activation, inhibition of the proteasome could also kill cancer cells by an alternative, non apoptotic form of cell death. We further demonstrated that proteasome inhibitors, such as MG132, could induce intracellular accumulation of polyubiquitinated proteins and extensive cellular vacuolization likely due to ER stress. Translational or transcriptional inhibitors suppressed MG132-induced polyubiquitinated protein accumulation, and in turn inhibited MG132-induced ER stress, cellular vacuolization and cell death. These findings thus suggested that non-apoptotic cell death was resulted from misfolded protein accumulation and ER stress. Furthermore, our study indicated that proteasome inhibitors could be favorable chemotherapeutic agents because they could induce non-apoptotic cell death in addition to apoptosis, which could overcome resistance due to compromised apoptotic machinery. PMID- 17912639 TI - The patient experience. AB - The impact of improved treatments for the management of hormone-sensitive breast cancer extends beyond clinical responses. Thanks to appropriate literature and access to the internet, patient awareness of treatment options has grown and patients are now, in many cases, able to engage their oncologists in informed conversations regarding treatment and what to expect in terms of efficacy and safety. Indeed, patients realize that although there is no cure for metastatic disease, treatment can greatly reduce the risk of progression and in the adjuvant setting, where treatment is administered with a curative intent, current treatment options reduce the risk of relapse. The approval of letrozole throughout the breast cancer continuum has provided patients with many reassuring options. The improvement in outcome with letrozole is achieved without a detrimental effect on overall quality of life. Adverse events such as hot flushes, arthralgia, vaginal dryness, and potential osteoporosis are most significant from the patient's perspective, and it is important that caregivers pay attention to patients experiencing these events, as they can impact compliance unless effectively explained and managed. The major benefits of letrozole are to improve prospects for long-term survivorship in the adjuvant setting and to delay progression and the need for chemotherapy in the metastatic setting. PMID- 17912640 TI - Femara and the future: tailoring treatment and combination therapies with Femara. AB - Long-term estrogen deprivation treatment for breast cancer can, in some patients, lead to the activation of alternate cellular pathways, resulting in the re emergence of the disease. This is a distressing scenario for oncologists and patients, but recent intensive molecular and biochemical studies are beginning to unravel these pathways, revealing opportunities for new targeted treatments. Far from making present therapies redundant, these new discoveries open the door to novel combination therapies that promise to provide enhanced efficacy or overcome treatment resistance. Letrozole, one of the most potent aromatase inhibitors, is the ideal candidate for combination therapy; indeed, it is one of the most intensively studied aromatase inhibitors in the evolving combinatorial setting. Complementary to the use of combination therapy is the development of molecular tools to identify patients who will benefit the most from these new treatments. Microarray gene profiling studies, designed to detect letrozole-responsive targets, are currently under way to understand how the use of the drug can be tailored more efficiently to specific patient needs. PMID- 17912642 TI - Interleukin-1beta enhances the effect of serum deprivation on rat annular cell apoptosis. AB - Excessive apoptosis of disc cells is believed to play an important role in intervertebral disc (IVD) degeneration. It has been shown that interleukin-1beta (IL-1beta) is involved in the failure of disc matrix by suppressing the synthesis of matrix components and stimulating the expression of matrix metalloproteinases. However, whether IL-1beta induces disc cell apoptosis is still unclear. The objective of this study was to investigate the effect of IL-1beta on the apoptosis of rat annular cells cultured with or without serum supplement. First passage rat annular cells were cultured with 0% or 10% fetal bovine serum (FBS) supplement and stimulated with 0, 10, 20 or 50 ng/ml IL-1beta for 12, 24 or 48 h. Apoptotic incidences were quantified by flow cytometry, morphologic changes in apoptotic cells were visualized by Hoechst 33258 staining and phase-contrast microscopy, and caspase-3 activity was also determined. When rat annular cells were cultured with 10% FBS supplement, no significant changes in apoptotic incidences, apoptotic morphology and caspase-3 activity were observed even when cells were stimulated with 50 ng/ml IL-1beta for 48 h. In contrast, serum deprivation for 24 h led to an increase in apoptotic incidences, the number of apoptotic nuclei and caspase-3 activity, and IL-1beta significantly increased the effects of serum deprivation in a dose-dependent manner. Our results indicate that IL-1beta alone is not a sufficient stimulus to induce disc cell apoptosis and that in order to suppress disc cell apoptosis, improving the nutrient supply to the disc may be more effective than antagonizing the adverse effects of IL 1beta. PMID- 17912643 TI - In vitro antioxidant and anti-rhizopus activities of Lamiaceae herbal extracts. AB - Eighty-eight extracts of different polarity obtained from 18 Lamiaceae medicinal and aromatic plants were screened for their antioxidant and antifungal properties. A TLC analysis combined with two bioassays were used to detect these activities. In most cases the highest radical scavenger capacity was detected in methanolic and aqueous extracts and polyphenols may be responsible. The most interesting antioxidant activity was observed in polar extracts obtained from Lycopus europaeus, Melissa officinalis, Origanum vulgare subsp. virens and Lavandula latifolia. On the contrary, the best results for the antifungal test against Rhizopus stolonifer were produced by non-polar herbal extracts. Phlomis lychnitis, Salvia pratensis and Calamintha sylvatica caused the highest inhibition on Rhizopus growth. PMID- 17912645 TI - [The bone and joint decade--chances for orthopedics and traumatic surgery]. PMID- 17912644 TI - Influences on practitioner treatment selection: best research evidence and other considerations. AB - The recent movement toward evidence-based practice in mental health services has highlighted the importance of research evidence in treatment decisions. However, the fact that many treatments with strong research support are not widely used in clinical settings suggests that practitioners' decisions are not based on research alone but rather are influenced by other considerations. This study examines the relative importance of various considerations on practitioner treatment selection using a national survey of mental health practitioners including doctoral-level psychologists, master's-level psychologists, and master's-level clinical social workers (N = 206). Results indicate that practitioners are influenced by a range of considerations including empirical evidence from applied field studies, the perceived flexibility of a treatment, and the appeal of a treatment to colleagues and clients. These findings are discussed within the context of efforts to design, evaluate, and disseminate treatments with research support into clinical settings. PMID- 17912647 TI - [As a clinical fellow at Brigham and Women's Hospital]. PMID- 17912646 TI - [Body weight and the effect of stationary conservative therapy for back pain]. PMID- 17912649 TI - [Aging patients profit from intensive care]. PMID- 17912648 TI - [Tantalum-implant for femoral head necrosis--an alternative?]. PMID- 17912659 TI - [Socioeconomic relevance of osteoarthritis in Germany]. AB - AIM OF STUDY: The aim of this study was to summarise the impact of medical care and related costs due to the treatment of osteoarthritis in Germany. MATERIAL AND METHODS: Data from different health care institutions, government authorities and public health insurances for the year 2002 were analysed. Amount and cost of acute and rehabilitation treatments (in-patient as well as out-patient), sickness leave and early retirement related to osteoarthritis were estimated. RESULTS: Advanced hip and knee osteoarthritis have a high socioeconomic impact in Germany. The number of people affected is growing, as is the overall incidence of joint replacement, especially knee arthroplasties and the percentage of in-patient rehabilitation for osteoarthritis. CONCLUSION: Our study confirms that the direct and indirect costs attributable to osteoarthritis are substantial and the resulting socioeconomic burden is significant. Since age is a major risk factor for osteoarthritis, the demographic changes will lead to an increased need for medical treatment of osteoarthritis patients in the future. PMID- 17912660 TI - [Is there a correlation between the clinical, radiological and intrasurgical findings of osteoarthritis of the knee? A Prospective study on 103 patients]. AB - AIM: The aim of this study was to compare the different diagnostic features (symptoms, physical examination, radiographic features and intraoperative findings) of osteoarthritis of the knee before total joint replacement and consequently relate them with each other. METHOD: Patients with primary osteoarthritis of the knee, who where accepted for total joint replacement in our clinic, were integrated in our study. The medical conditions were recorded by using the Womac score, the knee subscore (KS) of the Knee Society Clinical Rating System and a visual analogue scale (VAS). A physical examination was also performed. The radiological evaluation was performed by using standardised radiographs. Joint space narrowing, subchondral sclerosis, osteophytes, knee alignment and the Kellgren score were recorded. During surgery an orthopaedic specialist documented the progression of cartilage lesions using the classification of chondromalacia described by Outerbridge. The analysis of correlation was performed by using the Spearman correlation (SpK) coefficient. RESULTS: 103 patients were integrated in the study (mean age: 68 years, 70 women). No significant correlation could be found between the Womac score or VAS and the results of the radiological examination (SpK [Womac - Kellgren score]: - 0.04; SpK [VAS - Kellgren score]: 0.08). There was a significant correlation between the KS and the results of the radiological examination as well as the progression of the cartilage lesions, caused by the results of the physical examination documented in the KS (SpK [KS - Kellgren score]: - 0.39). The radiographic features, with the exception of subchondral sclerosis, showed a significant correlation with the degree of chondromalacia (SpK [Kellgren score - chondromalacia]: 0.43). The amount of osteophytes correlated the most with the cartilage lesions: SpK: 0.43. CONCLUSION: The knee subscore appears to be an efficient method for staging the clinical progression of osteoarthritis of the knee for clinical practice as well as for clinical trials. The Womac score especially serves to record the level of pain. We were able to confirm that the presence of osteophytes is the most significant radiographic feature of osteoarthritis of the knee. The Kellgren score turned out to be a reliable method for monitoring the radiographic progression of osteoarthritis of the knee. PMID- 17912661 TI - [Osteochondral fractures of the distal femur]. AB - BACKGROUND: Chondral or osteochondral avulsions of the lateral distal femur edge after luxation of the patella are well known. Less common are impression fractures of the retropatellar joint or the lateral trochlea. Furthermore, on standard knee X-rays these injuries may not be seen and thus not properly be treated. By presenting our cases we show clinical symptoms, diagnostic pathways and the therapeutic approach for osteochondral fractures of the distal femur. MATERIALS AND METHODS: The first patient had a large osteochondral impression fracture of the lateral femur trochlea and retropatellar surface. The second patient had a chondral depression of the lateral trochlea femoris after kicking a football. The further examinations showed no other knee injuries in the first case, but a rupture of the outer meniscus plus ACL rupture and distortion of the medial collateral complex in the other one. RESULTS: In both cases the whole degree of the injury was not seen on the standard X-rays, but only on MRI or CT scans. The clinical signs were also not so overwhelming, as both patients could walk with crutches, but suffered from haemarthrosis. Both patients were operated by arthroscopy first with surgery on the meniscus injury and ACL rupture. The impression was revised by an open procedure with elevation of the subchondral spongiosa. In the follow-up examination we saw no dissection of chondral flakes or local necrosis. CONCLUSION: Osteochondral lesions of the distal femur can be neglected, as it is an uncommon diagnosis and the radiological signs may not be impressing. The more important is a subtle anamnesis and further diagnostic regime via CT or MRI as it is a prearthrotic injury. Intraoperatively the impression must be elevated or in the case of a dislocation refixed. Further knee injuries have to be detected. In the follow-up one should check for signs of flake dissection or necrosis. PMID- 17912663 TI - [Computer-assisted open-wedge osteotomy]. AB - AIM: The open-wedge high tibial osteotomy is an established procedure for the treatment of the unicompartimental gonarthrosis in young patients. An adequate correction of osseous malalignment is crucial for a sufficient reduction of stress in a diseased compartment. We have examined reliability and precision of an intraoperatively used computer-assisted navigation system for high tibial osteotomy. The aim of the study is to show the equivalent safety and effectiveness of high tibial osteotomies carried out with the computer-assisted navigation system. It is assumed that a good correlation between the mechanical tibio-femoral axis as measured by radiography and by the navigation system can be achieved. METHOD: 40 medial open-wedge osteotomies were performed with computer assisted navigation on 39 patients (27 males, 12 females) between 1/2004 and 8/2006. The average age was 46.3 years (range: 26 - 64 years), the average weight was 83.2 kg (range: 54 - 118 kg). RESULTS: A good correlation between radiographic data and the data acquired with the navigation system was found for the tibio-femoral axis: preoperative data (8.0 +/- 2.5 degrees, radiographic; 7.8 +/- 2.1 degrees navigated) for varus alignment. The postoperative correlation was lower than the preoperative one (postoperative data (0.6 +/- 3.2 degrees radiographic; - 0.7 +/- 1.0 degrees navigated) for valgus alignment. In 2 patients a loss of correction occurred and had to be treated operatively. CONCLUSION: High tibial osteotomy is an established therapy procedure for unicompartmental gonarthrosis. It can be improved in its precision and reliability by computer-assisted navigation. PMID- 17912664 TI - [Analysis of 10-year survival after flexion osteotomy for femoral head necrosis]. AB - AIM: We have investigated the value of flexion osteotomy for femoral head necrosis with regard to the survival rate. METHOD: We examined 40 patients who were treated by flexion osteotomy for femoral head necrosis at the Hannover Medical School in the years 1969 to 1995. The mean follow-up period was 14.1 years. RESULTS: The mean survival time of the flexion osteotomies was 9.6 years. In 23 of the 40 patients, it failed within 10 years after surgery, which implies a 10-year survival rate of 42.5 %. For these 23 patients, the implantation of a total hip arthroplasty (THA) became necessary after an average time of 6.6 years. CONCLUSION: Due to the poorly predictable success of the flexion osteotomy and the comparatively poor patient satisfaction, the indication for flexion osteotomy in such cases should be reconsidered. PMID- 17912665 TI - [Polyethylene abrasion: cause or consequence of an endoprosthesis loosening? Investigations of firm and loosened hip implants]. AB - AIM: Periprosthetic tissue was analysed by the combination of different investigation techniques without destruction. The localisation and geometry of polyethylene abrasion particles were determined quantitatively to differentiate between abrasion due to function and abrasion due to implant loosening. Non polyethylene particles from implant components which contaminate the tissue were micro-analytically measured. The results will help us to understand loosening mechanisms and thus lead to implant optimisations. METHOD: A non-destructive particle analysis using highly sensitive proton-induced X-ray emission (PIXE) was developed to achieve a better histological allocation. Five autopsy cases with firmly fitting hip endoprosthesis (2 x Endo-Modell Mark III, 1 x St. Georg Mark II, LINK, Germany; 2 x Spongiosa Metal II, ESKA, Germany) were prepared as ground tissue specimens. Wear investigations were accomplished with a combined application of different microscopic techniques and microanalysis. The abrasion due to implant loosening was histologically evaluated on 293 loosened cup implants (St. Georg Mark II, LINK, Germany). RESULTS: Wear particles are heterogeneously distributed in the soft tissue. In cases of cemented prostheses, cement particles are dominating whereas metal particles could rarely be detected. The concentration of the alloy constituent cobalt (Co) is increased in the mineralised bone tissue. The measured co-depositions depend on the localisation and/or lifetime of an implant. Functional polyethylene (PE) abrasion needs to be differentiated from PE abrasion of another genesis (loosening, impingement) morphologically and by different tissue reactions. CONCLUSION: In the past a reduction of abrasion was targeted primarily by the optimisation of the bearing surfaces and tribology. The interpretation of our findings indicates that different mechanisms of origin in terms of tissue contamination with wear debris and the alloy should be included in the improvement of implants or implantation techniques. PMID- 17912667 TI - [The stabilizing effect of atmospheric pressure (AP) on hip joint subject to traction force--an experimental study]. AB - AIM: We aimed to prove the stabilising effect of atmospheric pressure (AP) on the hip joint experimentally. METHOD: In the experiment, model joints of 28 mm, 32 mm und 36 mm diameter were subjected to increasing traction force. The acting force and the resulting dislocation distance were measured both with the capsule hermetically sealed, as well as with the capsula open. RESULTS: For the hermetically sealed capsule we measured maximum resistances of 7.6 kp for the 28 mm joint, 10.4 kp for the 32 mm joint and 12.4 kp for the 36 mm joint. With the capsule open we found resistances from 0.4 kp to 1 kp. Our experimental results exceeded the predicted resistances of 6 kp, 7.8 kp and 9.9 kp. Increased amounts of synovial fluid reduced the stability. CONCLUSION: Our measurements confirm the continual stabilising effect of AP on the hip joint, which can be quantified as the resting potential of stability (RPS) or luxation work (LW). The RPS is calculated by multiplying the difference of AP and saturated vapour pressure of synovial fluid with the cross-sectional area of the femoral head. It represents the force, necessary for luxation of the joint against the resistance of AP. The RPS is proportional to the square of the joint diameter. The LW, calculated by multiplying RPS with the luxation distance, is proportional to the joint diameter cubed. That is why a small increase of joint diameter leads to a significant increase of stability, while the rate of the increase of range-of-motion decreases. To achieve stability of a total hip arthroplasty the size of the joint components should depend on the size of the resected femoral head. Also the hermetically sealed capsule should be reconstructed carefully. PMID- 17912666 TI - [Durom hip resurfacing arthroplasty: first clinical experiences with a lateral approach]. AB - INTRODUCTION: We present the first clinical and radiographic results of the Durom Hip Resurfacing prosthesis with a lateral approach. MATERIALS AND METHODS: 52 patients (n = 59 protheses) with a mean follow-up of 25.4 +/- 10 month were evaluated. Clinical evaluation was done using the Harris hip score (HHS), a modified University of California at Los Angeles (UCLA) Activity Level Scale and the Merle d'Aubigne Score pre- and postoperatively. Radiographic evaluation included the preoperative femoral neck-shaft angle, the pre- and postoperative femoral offset, the postoperative stem-shaft angle as well as postoperative periprosthetic radiolucencies. RESULTS: Within the clinical follow-up the postoperative mean HHS (93 +/- 4.3 vs. 41.2 +/- 7.1 points), the modified UCLA (8.8 +/- 2.8 vs. 4.8 +/- 1.9) as well as the Merle d'Aubigne Score (17.1 +/- 1.7 vs. 7.5 +/- 2.1) indicated a significant improvement (p or = 50 mg kg(-1) of microencapsulated fenobucarb had a barrier effect within a single day, and could stop or retard the penetration of termites for 7 days. Microencapsulated fenobucarb did not act as a repellent, as indicated by a gradual increase in mortality at > or = 50 mg kg(-1) with slight penetration into the treated soil during the 7 day test. CONCLUSION: The results suggest that microencapsulated fenobucarb is a fast-acting termiticide, with a good barrier effect as a soil treatment, that also acts as a reduced repellent, retarding entry of termites into treated soil. PMID- 17912686 TI - Biological control of terrestrial molluscs using Phasmarhabditis hermaphrodita- progress and prospects. AB - Phasmarhabditis hermaphrodita Schneider (Nematoda: Rhabditidae) is a nematode that parasitises a wide range of slug and snail species. It has been formulated into a biological control agent (Nemaslug) and was commercialised in 1994. It is now available in fourteen European countries. A review is given of all research on P. hermaphrodita, including basic biology, mass cultivation, formulation, host range, application strategies, field efficacy and effects on non-target organisms. The many critical gaps in present knowledge are highlighted, and future research is proposed that will lead to greater understanding of this unusual parasite and may enable its more widespread use in the management of mollusc pests. PMID- 17912687 TI - The emerging field of chemical genetics: potential applications for pesticide discovery. AB - The use of small molecules to probe biological systems, generally described as 'chemical genetics', has grown considerably in the past 7 years, especially in areas related to human biology and therapeutics. This review describes some aspects of chemical genetics technologies that can be usefully applied to pesticide target discovery and lead generation. The chemical genetics approach (consisting of a phenotype screen, a chemical library and a robust target identification methodology) is compared with conventional and target-based screening. The outcomes of a chemical genetics approach are novel protein targets coupled with in vivo-active chemical ligands. The 'chemistry-first' paradigm of the chemical genetics approach can circumvent some of the obstacles that have emerged for the exploitation of novel but chemically unvalidated targets identified from genetic or genomic screens. Some of the advantages and challenges in using chemical genetics approaches are reviewed. PMID- 17912688 TI - The effect of dextromethorphan in preventing cholecalciferol-induced poison shyness and sickness-induced anorexia in the laboratory Norway rat. AB - BACKGROUND: Overcoming bait and poison shyness is critical to the success of pest control operations against rats and other rodents. The authors hypothesized that the N-methyl-D-aspartate receptor blocker, dextromethorphan, could prevent the acquired memory of sickness and sickness-induced anorexia resulting from rodents eating poisoned bait. RESULTS: Cholecalciferol (1/4 LD(50)) was mixed with dextromethorphan and fed to rats on two 2 day sessions, with an 18 day break in between. Dextromethorphan did not prevent poison shyness; during the second poisoning period, both the cholecalciferol only and the cholecalciferol plus dextromethorphan groups had lower intakes of the bait compared with the control and dextromethorphan only groups. In addition to the previously recorded symptoms of cholecalciferol poisoning, the rats in this trial were observed to have nose bleeds, weepy eyes, laboured breathing and, in the case of the cholecalciferol only treated group, a period of decreased water intake followed by a period of increased water intake. There was also a period of increased water intake in the cholecalciferol plus dextromethorphan group. CONCLUSION: Dextromethorphan failed to prevent poison shyness and the anorectic effect of cholecalciferol. However, it did reduce anorexia from 17 days in the cholecaliferol group to 8 days in the cholecalciferol plus dextromethorphan group. PMID- 17912689 TI - Antiphytoviral activity of bruceine-D from Brucea javanica seeds. AB - BACKGROUND: Brucea javanica (L.) Merr. is widely distributed throughout the southern parts of China and has been used in traditional medicine to treat a variety of diseases. The objective of the present study was to identify the active antiphytoviral compound in the seeds of B. javanica and evaluate the inhibitory activity of the compound against plant virus. RESULTS: Bioassay-guided fractionation of the most active extract from the seeds led to the isolation of an antiphytoviral compound which was identified as bruceine-D by conventional spectroscopy methods. The compound exhibited significant inhibitory activity against the infection and replication of tobacco mosaic virus (TMV), with IC(50) values of 13.98 and 7.13 mg L(-1) respectively. The compound also showed a strong inhibitory effect on the infectivity of potato virus Y (PVY) and cucumber mosaic virus (CMV). Furthermore, the compound could effectively inhibit systemic TMV infection in the host tobacco plant under glasshouse conditions. CONCLUSION: The results suggested that bruceine-D from Brucea javanica may have the potential to be used as a natural viricide, or a lead compound for new viricides. PMID- 17912690 TI - Expression of garlic leaf lectin under the control of the phloem-specific promoter Asus1 from Arabidopsis thaliana protects tobacco plants against the tobacco aphid (Myzus nicotianae). AB - BACKGROUND: To check for correlation between the insecticidal properties and the specificity of lectins, a comparative study was made of the insecticidal activities of two garlic lectins with different biological activities. RESULTS: The insecticidal activity of the garlic (Allium sativum L.) leaf lectin ASAL and bulb lectin ASAII towards the tobacco aphid Myzus nicotianae Blackman was studied using bioassays with transgenic tobacco (Nicotiana tabacum L. cv. Wisconsin 38). Bioassays were started with newborn nymphs of the tobacco aphid. Although during the first 7-8 days when nymphs developed to adults there were no apparent effects, part of the nymphal population was found to develop into winged (alate) forms. Later it became clear that transgenic plants expressing ASAL and ASAII had a significant effect on the reproduction capacity of the resulting adults, with a reduction of up to 40%. Different life table parameters such as prereproductive time, intrinsic rate of natural increase, generation time and doubling time were significantly affected (P < 0.05) in aphids grown on transgenic plant material expressing ASAL and ASAII. CONCLUSION: Bioassays with tobacco plants expressing ASAL and ASAII demonstrated a significant impact on the population growth of M. nicotianae. Therefore, both lectins can be considered as valuable candidate aphid control agents. PMID- 17912691 TI - Susceptibility of lesser mealworm (Coleoptera: Tenebrionidae) adults and larvae exposed to two commercial insecticides on unpainted plywood panels. AB - BACKGROUND: The susceptibilities of adult and larval lesser mealworms, Alphitobius diaperinus (Panzer), to two commercially formulated insecticides, cyfluthrin and tetrachlorvinphos, were examined through exposure on treated plywood panels. Lesser mealworms were collected from four caged-layer poultry farms, three in New York and one in Maine. An additional strain was obtained from an infestation occurring in a cricket colony. RESULTS: In all poultry farm derived strains, a portion of the population (1.8-16.2%) survived cyfluthrin exposure. The Maine and cricket colony strains were tolerant of tetrachlorvinphos exposure as both larvae and adults, with 55-74% mortality, whereas nearly 100% mortality was observed with New York strains. The cricket colony adult beetles were highly susceptible to cyfluthrin, with 100% mortality following exposure, but larvae were considerably less susceptible (87.7%). Pesticide use histories for the poultry farms and their impact on the results are discussed. CONCLUSION: The results document that tetrachlorvinphos, an active ingredient with a long use history, may be losing its effectiveness against lesser mealworms in some poultry operations; however, it is still effective in many others. PMID- 17912692 TI - Metabolism of imidacloprid and DDT by P450 CYP6G1 expressed in cell cultures of Nicotiana tabacum suggests detoxification of these insecticides in Cyp6g1 overexpressing strains of Drosophila melanogaster, leading to resistance. AB - BACKGROUND: With the worldwide use of insecticides, an increasing number of pest insect species have evolved target-site or metabolism-based resistance towards some of these compounds. The resulting decreased efficacy of pesticides threatens human welfare by its impact on crop safety and further disease transmission. Environmental concentrations of some insecticides are so high that even natural populations of non-target, non-pest organisms such as the fruit fly Drosophila melanogaster Meig. have been selected for resistance. Cyp6g1-overexpressing strains of D. melanogaster are resistant to a wide range of chemically diverse insecticides, including DDT and imidacloprid. However, up to now there has been no evidence that the CYP6G1 enzyme metabolises any of these compounds. RESULTS: Here it is shown, by heterologous expression in cell suspension cultures of Nicotiana tabacum L. (tobacco), that CYP6G1 is capable of converting DDT (20 microg per cell culture assay) by dechlorination to DDD (18% of applied amount in 48 h), and imidacloprid (400 microg) mainly by hydroxylation to 4 hydroxyimidacloprid and 5-hydroxyimidacloprid (58 and 19% respectively in 48 h). CONCLUSION: Thus, the gap between the supposed resistance gene Cyp6g1 and the observed resistance phenomenon was closed by the evidence that CYP6G1 is capable of metabolising at least two insecticides. PMID- 17912693 TI - Delaying weed adaptation to herbicide by environmental heterogeneity: a simulation approach. AB - BACKGROUND: Environmental heterogeneity in space or time can drive the evolutionary trajectory of an adaptive trait. This concept could be of practical significance in pesticide resistance management that aims to delay the evolution of a resistance allele. Using a population genetics model, the dynamics of herbicide resistance in a weed species was simulated in a heterogeneous environment with alternation of two unrelated herbicides in time, in space or in both time and space. The level of the environmental heterogeneity (habitat grain) was simulated by a variation in the size of the herbicide-treated areas. RESULTS: The model confirms that several strategies based on habitat heterogeneity efficiently slow down and even prevent resistance evolution. For a recessive resistant trait in outcrossing species, a medium level of environmental heterogeneity (intermediate habitat grain) was found to be the best for delaying herbicide resistance, as previously observed for insecticide resistance management. In selfing species or for a dominant resistant trait in outcrossing species, a low level of environmental heterogeneity (coarse-grained habitat) was more efficient in delaying resistance evolution when heterogeneity in both space and time were considered. CONCLUSION: This model suggests that the choice of optimal tactics for delaying herbicide resistance by enhanced heterogeneity in space or time firstly depends on the interactions between the breeding system and the dominance of the resistance allele in the presence of herbicide, then on the value of the fitness cost and lastly on the dominance of this fitness cost. PMID- 17912694 TI - The management of Cesarean scar ectopic pregnancy following treatment with methotrexate--a clinical challenge. AB - We present a case of Cesarean scar ectopic pregnancy, complicated by the persistence of clinical symptoms despite a rapid and complete biochemical response to a single systemic injection of methotrexate. A 34-year-old woman with three previous Cesarean sections was diagnosed with a Cesarean scar ectopic pregnancy following in-vitro fertilization treatment. The diagnosis was suggested by three-dimensional (3D) ultrasound scan and confirmed with magnetic resonance imaging (MRI). Management involved administration of a single systemic injection of methotrexate and follow-up with serial ultrasound assessments and serum beta human chorionic gonadotropin (beta-hCG) measurements. The main challenge was the persistence of clinical symptoms despite adequate medical treatment, as judged by complete resolution of biochemical trophoblastic activity, which resulted in repeated admissions to the hospital. Serial transvaginal ultrasound scans showed an initial increase in the size of the mass, which led to increasing anxiety in the couple. Eventually, 15 weeks after the administration of methotrexate, the couple requested surgical intervention. An uneventful surgical resection of the abnormal area, which showed appearances suggestive of trophoblastic tissue, was undertaken to good effect. In summary, despite a rapid normalization of serum beta-hCG following the administration of methotrexate, the patient remained symptomatic and had ultrasound appearances suggestive of incomplete resorption of trophoblast, necessitating surgical intervention. PMID- 17912695 TI - Evaluation of the antimicrobial activity of ebselen: role of the yeast plasma membrane H+-ATPase. AB - Ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one) is a selenium-containing antioxidant demonstrating anti-inflammatory and cytoprotective properties in mammalian cells and cytotoxicity in lower organisms. The mechanism underlying the antimicrobial activity of ebselen remains unclear. It has recently been proposed that, in lower organisms like yeast, the plasma membrane H+-ATPase (Pma1p) could serve as a potential target for this synthetic organoselenium compound. Using yeast and bacteria, the present study found ebselen to inhibit microbial growth in a concentration- and time-dependent manner, and yeast and Gram-positive bacteria to be more sensitive to this action (IC50 approximately 2-5 microM) than Gram-negative bacteria (IC50 < 80 microM). Washout experiments and scanning electron microscopic analysis revealed ebselen to possess fungicidal activity. In addition, ebselen was found to inhibit medium acidification by PMA1-proficient haploid yeast in a concentration-dependent manner. Additional studies comparing PMA1 (+/-) and PMA1 (+/+) diploid yeast cells revealed the mutant to be more sensitive to treatment with ebselen than the wild type. Ebselen also inhibited the ATPase activity of Pma1p from S. cerevisiae in a concentration-dependent manner. The interaction of ebselen with the sulfhydryl-containing compounds L cysteine and reduced glutathione resulted in the complete and partial prevention, respectively, of the inhibition of Pma1p ATPase activity by ebselen. Taken together, these results suggest that the fungicidal action of ebselen is due, at least in part, to interference with both the proton-translocating function and the ATPase activity of the plasma membrane H+-ATPase. PMID- 17912697 TI - Gene expression analysis of estrogenic compounds in the liver of common carp (Cyprinus carpio) using a custom cDNA microarray. AB - Exposure to a variety of compounds with estrogenic activity has been shown to interfere with normal developmental and reproductive processes in various vertebrate species. The aim of this study was to determine the transcriptional profile of the natural estrogen, 17 beta-estradiol, and three synthetic estrogenic compounds (4-nonylphenol, bisphenol A, ethinylestradiol) in the liver of common carp, using a custom cDNA microarray. For that purpose, fish were aqueously exposed to three concentrations of each chemical for 24 or 96 h. Microarray analysis revealed that a total of 185 different gene transcripts were differentially expressed following exposure to at least one of the estrogen( like) concentrations. We were able to identify a common set of 28 gene fragments, whose expression was significantly modified in the same way by the three xenoestrogens and 17 beta-estradiol. Although several of these gene expression effects corroborated past literature data, we also discovered some novel target genes of (xeno)estrogen exposure, providing interesting insights into the molecular basis of estrogenic effects. In addition, each of the four compounds induced gene expression changes that were not, or only partially, shared by the other chemicals, suggesting that not all chemicals with estrogenic activity act alike. These results demonstrate the potential of our custom Cyprinus carpio microarray to detect common estrogen-like activity as well as to identify unique compound-associated effects of (estrogenic) endocrine disruptors in fish. PMID- 17912696 TI - Modulation of indomethacin-induced gastric injury by spermine and taurine in rats. AB - This study investigated the involvement of neutrophil infiltration, nitric oxide (NO) generation, and oxidative stress in indomethacin-induced ulcer and the possible gastroprotective potentials of spermine and taurine, known for their tissue regenerating and antioxidant effects, respectively. Male Wistar albino rats (180-220 g) were allocated into a normal control group, ulcer control group (received a single dose of indomethacin 40 mg-kg p.o.), and two ulcer groups pretreated with spermine (150 mg-kg p.o. 1 h before ulcer induction) and taurine (250 mg-kg i.p. for three consecutive days before ulcer induction). The animals were killed 6 h after indomethacin administration, and the gastric juice, serum, and mucosal tissue were used for gastric injury evaluation. Both modulators significantly ameliorated the indomethacin-induced gastric lesions in glandular mucosa. Notably, spermine exhibited the most pronounced effect as manifested by great reduction in the gastric ulcer index, normalization of the elevated gastric acidity, and triggering of mucin production. Spermine and taurine were able to decrease the elevated levels of gastric myeloperoxidase, conjugated diene, and serum NO. However, the lowered tissue NO content was markedly elevated only by taurine. The antioxidant action of taurine was illustrated by restoration of the depressed content of glutathione, normalization of the inhibited activities of glutathione reductase, and superoxide dismutase. These results suggest that spermine and taurine confer significant gastroprotection against indomethacin induced gastric injury with the priority of spermine. PMID- 17912698 TI - Using a comparative in vivo DNase I footprinting technique to analyze changes in protein-DNA interactions following phthalate exposure. AB - Exposure to environmental chemicals often induces changes in gene expression leading to a variety of developmental and physiological problems. Understanding the underlying mechanism of these changes will aid in assessing human risk to these chemicals. Traditional methods for analyzing protein-DNA interactions include in vivo footprinting and chromatin immunoprecipitation (ChIP). However, ChIP does not provide binding location, and conventional footprinting is too subjective and time consuming for comparing protein binding in toxicological studies. Here, in vivo DNase I footprinting is adapted for use with the automated DNA sequencer to provide a semiquantitative map of changes in DNA-protein interactions in the promoter of steroidogenic acute regulatory (StAR) protein. StAR is the rate-limiting step in testosterone biosynthesis and is downregulated following in utero di-butyl phthalate (DBP) treatment in rats through an unknown mechanism. In vivo footprinting identified three regions of altered DNase digestibility following DBP treatment, and EMSA identified the corresponding transcription factors as SF-1, c/ebp beta, and GATA4. ChIP assays confirmed changes in protein-binding activity of SF-1 and c/ebp beta, but only c/ebp beta gesponds to only DBP. This suggests that c/ebp beta ginding is involved in DBP induced transcriptional changes. By tailoring in vivo footprinting for toxicological studies, it can provide a detailed and accurate map of protein-DNA interactions and is an excellent first step in determining the changes in the structure of transcriptional machinery following an exogenous chemical treatment. PMID- 17912700 TI - Endothelial cytoprotection from oxidized LDL by some crude Melanesian plant extracts is not related to their antioxidant capacity. AB - Habitual consumption of some Melanesian medicinal and food plants may influence atherosclerosis development via their antioxidant capacity at the endothelial level. Areca nut (AN; Areca catechu), piper inflorescence (PBI; Piper betle), betel quid (BQ), guava buds (GB; Psidium guajava), the leaves (NL), juice (NJ), fruit (NF), and root (NR) of noni (Morinda citrifolia), the propagules of raw (MBR), and cooked (MBC) mangrove (Bruguiera gymnorrhiza) were evaluated for their ability to scavenge the 1,1-diphenyl-2-picryl-hydrazyle (DPPH) radical, to protect human low-density lipoprotein (LDL) from Cu2+-catalyzed oxidation and to protect cultured bovine aortal endothelial cells (BAEC) from oxidized LDL (oxLDL) induced cytotoxicity. Polyphenol-rich extracts AN, PBI, and BQ were potent DPPH scavengers, having similar activity to quercetin and able to protect LDL from oxidation in a dose-dependent manner at concentrations higher than 10 microg/mL, but were pro-oxidants at lower concentrations. These extracts were cytotoxic to BAEC at concentrations above 10 microg/mL and were unable to prevent oxLDL endotheliopathy. GB and NR at 10 mug/mL displayed both the ability to delay LDL oxidation and prevent oxLDL cytotoxicity, although the latter lacked the ability to scavenge the DPPH radical. At higher concentrations, however, both were cytotoxic in themselves. The remaining noni extracts NF, NJ, NL, and both mangrove extracts MBC and MBR were unable to protect LDL from oxidation at all tested concentrations, but were effective cytoprotective agents at 50 microg/mL. All extracts were able to prevent an oxLDL-mediated increase in intracellular aldehyde generation but had little effect on extracellular peroxidation as measured by thiobarbituric acid reactive substances (TBARS). On the basis of this model system, we conclude that the antioxidant benefits of AN, PBI, and BQ may be offset by their enhancement of their cytotoxic effects of oxLDL toward BAEC, whereas GB and low concentrations of noni and mangrove may be considered antiatherogenic. The discrepancies between our in vitro and cellular culture experiments emphasize the importance of experimental conditions in evaluating the antioxidant potential of crude plant extracts. PMID- 17912702 TI - Dimethylsphingosine and FTY720 inhibit the SK1 form but activate the SK2 form of sphingosine kinase from rat heart. AB - Fractionation of cytosolic sphingosine kinase (SKase) activity by gel filtration chromatography gave rise to a 96-kDa peak that contained only the SK2 form of SKase (by Western analysis) and a broad ca. 46 kDa peak that contained only SK1 forms. SK2 appeared to have a bound accessory protein. When tested with the classic SKase inhibitor dimethylsphingosine (DMS), SK1 was extensively inhibited; however, SK2 was not inhibited but unexpectedly was activated. Activation of SK2 was the result of DMS enhancing the affinity of the enzyme for sphingosine, and, at low concentrations of ATP and sphingosine, activated by more than 100%. Activation of SK2 could be demonstrated in the cytosolic fraction indicating it was unrelated to the purification step. The immunomodulator FTY720 also activated SK2 (although to a lesser extent), but was a potent inhibitor of SK1. SK2 from rat liver and spleen was also not inhibited by DMS. L-Sphingosine and to a lesser extent dihydrosphingosine and phytosphingosine were effective inhibitors of both forms. PMID- 17912701 TI - Lead-induced alterations of apoptosis and neurotrophic factor mRNA in the developing rat cortex, hippocampus, and cerebellum. AB - Previous reports have recently shown the prototypic neurotoxicant, lead, to induce apoptosis in the brains of developing organisms. In the current study, timed-pregnant rats were exposed to lead acetate (0.2% in the drinking water) 24 h following birth at postnatal day 1 (PND 1). Dams and pups were continuously exposed to lead through the drinking water of the dam until PND 20. Postnatal exposure in the pups resulted in altered mRNA levels of the following apoptotic and neurotrophic factors: caspase 2 and 3, bax, bcl-x, brain-derived neurotrophic factor (BDNF). Ribonuclease protection assays were conducted to measure the factors simultaneously at the following postnatal time points: 9, 12, 15, 20, 25, days. Our results suggest a brain region- and time-specific response following lead acetate exposure. The region most vulnerable to alterations occurs in the hippocampus with alterations beginning at PND 12, in which caspase 3, bcl-x, BDNF increase with lead exposure. Significant treatment effects were not observed for both the cortex and cerebellum. PMID- 17912703 TI - Transcriptional responses of alpha- and rho-class glutathione S-transferase genes in the liver of three freshwater fishes intraperitoneally injected with microcystin-LR: relationship of inducible expression and tolerance. AB - Rho-class glutathione S-transferase (GST) is found only in teleost fish with no homologues in mammals. Silver carp (Hypophthalmichthys molitrix), grass carp (Ctenopharyngodon idellus), and Nile tilapia (Oreochromis nilotica) are three warm freshwater fishes with differential tolerance to microcystin-LR (MC-LR): Nile tilapia has a little higher tolerance than silver carp, but both have much higher tolerance than grass carp. Full-length cDNAs encoding the rho-class GST were cloned and sequenced from the livers of the three fishes. The silver carp, grass carp, and Nile tilapia rho-class GST cDNAs were 1078, 1104, and 904 bp in length, respectively, and all contained an open-reading frame (ORF) of 681 bp (encoding a polypeptide of 226 amino acids). Using beta-actin as an external control, semiquantitative RT-PCR was conducted to determine the constitutive and inducible expression level of alpha- and rho-class GST genes among the three fishes (6-12 g) intraperitoneally injected with MC-LR (50 mug kg(-1) body weight). Liver mRNA expression levels of alpha-class GST gene were found to be higher than those of rho-class GST gene in both exposed and control fish of silver carp and grass carp, whereas liver mRNA expression levels of rho-class GST gene were higher than those of alpha -class GST gene in both exposed and control fish of Nile tilapia. Increased liver transcription of GST isoforms was detected at 8-h postinjection of MC-LR in silver carp (alpha- and rho-class GST) and Nile tilapia (rho-class GST), and at 24-h postinjection in silver carp (alpha-class GST) and Nile tilapia (alpha-class GST), but an increase in liver transcription neither of alpha-class GST nor of rho-class GST was detected in grass carp at either 8-h or 24-h postinjection. The inducible expression of the liver GST gene showed a close relationship with their tolerance to MC-LR: high-resistant fish (phytoplanktivorous silver carp and Nile tilapia) had inducible liver expression of either alpha- or rho-class GST, and the high-sensitive fish (herbivorous grass carp) had no inducible liver expression of either one. We suggest that inducible expression (instead of constitutive expression) of the liver GST gene should play an important role in the tolerance to microcystin exposure, and that in addition to alpha-class GST, high-liver expression of rho-class GST gene might have facilitated the evolutionary radiation of teleost fish. PMID- 17912705 TI - Baby steps: determining the relationship between Crohn's activity and birth outcomes. PMID- 17912710 TI - The unwary purchaser: consumer psychology and the regulation of commerce in America. AB - Starting in the 1870s, American jurists deciding cases of trademark infringement began advancing arguments that the ordinary purchaser was an unwary one, easily deceived by imitations. Embedded within their legal decisions was a vision of the typical consumers' habitual behavior and cognitive ability. In response to legal critics who argued that the presumed psychology of the consumer was unevenly deployed, applied psychologists developed laboratory-based experiments and scales for determining the likelihood that the "average" purchaser would be confused. Although these psychologists failed in their goal of securing regular legal patronage, this commercial context and the resulting experiments were constitutive of the delineation of "recognition" as a distinct mental process. Furthermore, this case study complicates the scholarly consensus about the role of standardization and personal responsibility in the liberal administration of mass society. PMID- 17912704 TI - Cigarette smoke extract affects functional activity of MRP1 in bronchial epithelial cells. AB - Cigarette smoke is the principal risk factor for development of chronic obstructive pulmonary disease (COPD). Multidrug resistance-associated protein 1 (MRP1) is a member of the ATP-binding cassette (ABC) superfamily of transporters, which transport physiologic and toxic substrates across cell membranes. MRP1 is highly expressed in lung epithelium. This study aims to analyze the effect of cigarette smoke extract (CSE) on MRP1 activity. In the human bronchial epithelial cell line 16HBE14o-, MRP1 function was studied flow cytometrically by cellular retention of carboxyfluorescein (CF) after CSE incubation and MRP1 downregulation by RNA interference (siRNA). Cell survival was measured by the MTT assay. Immunocytochemically, it was shown that 16HBE14o(-) expressed MRP1 and breast cancer resistance protein. Coincubation of CSE IC50 (1.53% +/- 0.22%) with MK571 further decreased cell survival 31% (p, = 0.018). CSE increased cellular CF retention dose dependently from 1.7-fold at 5% CSE to 10.3-fold at 40% CSE (both p < 0.05). siRNA reduced MRP1 RNA expression with 49% and increased CF accumulation 67% versus control transfected cells. CSE exposure further increased CF retention 24% (p = 0.031). A linear positive relation between MRP1 function and CSE-modulating effects (r = 0.99, p =0.089) was shown in untransfected, control transfected, and MRP1 downregulated 16HBE14o- cells analogous to blocking effects with MRP1 inhibitor MK571 (r = 0.99, p = 0.034). In conclusion, cigarette smoke extract affects MRP1 activity probably competitively in bronchial epithelial cells. Inhibition of MRP1 in turn results in higher CSE toxicity. We propose that MRP1 may be a protective protein for COPD development. PMID- 17912712 TI - From craniology to serology: racial anthropology in interwar Hungary and Romania. PMID- 17912714 TI - Baring the soul: Paul Bindrim, Abraham Maslow and 'nude psychotherapy'. AB - Nude psychotherapy is one of the most flamboyant therapeutic techniques ever developed in American psychology. Largely forgotten today, the therapy was an academic and popular sensation upon its introduction in 1967. Developed by psychologist Paul Bindrim, the therapy promised to guide clients to their authentic selves through the systematic removal of clothing. This paper explores the intellectual, cultural and ethical context of nude therapy and its significance as a form of unchurched spirituality. Although nude therapy has an indisputable tabloid character, it is also rooted in a long-standing academic search for authenticity and ultimate meaning through science. Bindrim's career demonstrates the historically long-standing interweaving of spirituality and science within American psychology while simultaneously highlighting the field's extraordinary capacity for adaptive reinvention. PMID- 17912716 TI - Science in Dachau's shadow: Hebb, Beecher, and the development of CIA psychological torture and modern medical ethics. PMID- 17912721 TI - Vasculoprotective potential of olive oil components. AB - Epidemiological and clinical studies found that the traditional Mediterranean style diet is associated with significantly lower mortality from coronary artery disease. Although it is difficult to isolate individual dietary factors, cumulative evidence suggests that olive oil, used as primary source of fat by Mediterranean populations, may play a key role in the observed cardiovascular benefit. Olive oil is a priceless source of vitamins and polyphenolic antioxidants, and has a balanced ratio of monounsaturated and polyunsaturated fatty acids. There are multiple mechanisms by which olive oil might impact the development of atherosclerosis. Olive oil decreases LDL-cholesterol and increases HDL-cholesterol, and also reduces oxidative stress due to polyphenols, which are able to scavenge free radicals and protect LDL from oxidation. In addition, olive oil components may interfere with the inflammatory response within atherosclerotic lesion, by inhibiting endothelial activation involved in monocyte recruitment during early atherogenesis and macrophage production of inflammatory cytokines and matrix degrading enzymes, thus improving vascular stability. Other vasculoprotective mechanisms by olive oil components derive from anti-thrombotic and anti-hypertensive actions. The available data support the need to preserve certain dietary traditions, such as olive oil consumption, to counteract the burden of cardiovascular disease. PMID- 17912722 TI - The potential role of olive oil-derived MUFA in insulin sensitivity. AB - Dietary fatty acids play an important role in the development of insulin resistance, the prelude to type 2 diabetes mellitus. This review addresses the potential role of olive oil-derived MUFA in insulin sensitivity, particularly how dietary fat interacts with insulin resistance looking at whole body metabolic measures, as well as molecular effects. The review focuses on the role of non esterified fatty acids, fatty acid composition in vivo and dietary fat modification on insulin resistance in the metabolic syndrome. Particular emphasis is placed on the role of olive oil within the context of dietary modification to improve insulin sensitivity and for the prevention of the metabolic syndrome. PMID- 17912723 TI - Obesity and the metabolic syndrome in Mediterranean countries: a hypothesis related to olive oil. AB - In Mediterranean countries people would previously have consumed a diet with a high proportion of MUFA. Physical activity would have been intense with a low level of stress. The stearoyl-CoA desaturase (SCD1) system selected over thousands of years of this type of behavior must have adapted to a particular capacity of self regulation. Now, this pattern, called the "Mediterranean diet", has been broken and many people living by the Mediterranean consume a high quantity of calories, mainly from saturated or n-6-rich fats and the relative intake of MUFA has decreased. Simultaneously, physical activity has decreased and the pattern of stress has changed towards what is called a western lifestyle. In this new context, if people have a favorable, genetically conditioned SCD1 activity that will let them confront the new situation or else have some other compensatory mechanism, such as being keen on sport, etc, then they can prevent the appearance of some of the complications associated with the metabolic syndrome. If, on the other hand, the SCD1 pattern is genetically unfavorable for this new situation and they have a new cultural context, then they do not have the alternative compensatory mechanisms and the probability of developing the metabolic syndrome is high. PMID- 17912724 TI - Alternate-site enzyme promiscuity. PMID- 17912725 TI - From 2,4-diphospha-3-thia- and -3-selenapentadienes [(Me3Si)2C=P]2E to heteronorbornane cage compounds. PMID- 17912726 TI - Molecular cosensitization for efficient panchromatic dye-sensitized solar cells. PMID- 17912727 TI - Plasmon-driven synthesis of triangular core-shell nanoprisms from gold seeds. PMID- 17912728 TI - Bulky chiral carbene ligands and their application in the palladium-catalyzed asymmetric intramolecular alpha-arylation of amides. PMID- 17912729 TI - Tandem catalysis for a one-pot regioselective protection of carbohydrates: the example of glucose. PMID- 17912730 TI - Chiral symmetry breaking by chemically manipulating statistical fluctuation in crystallization. PMID- 17912732 TI - Selective catalytic oxidation of ethanol to acetic acid on dispersed Mo-V-Nb mixed oxides. AB - The direct oxidation of ethanol to acetic acid is catalyzed by multicomponent metal oxides (Mo-V-NbO(x)) prepared by precipitation in the presence of colloidal TiO(2) (Mo(0.61)V(0.31)Nb(0.08)O(x)/TiO(2)). Acetic acid synthesis rates and selectivities (~95 % even at 100 % ethanol conversion) were much higher than in previous reports. The presence of TiO(2) during synthesis led to more highly active surface areas without detectable changes in the reactivity or selectivity of exposed active oxide surfaces. Ethanol oxidation proceeds via acetaldehyde intermediates that are converted to acetic acid. Water increases acetic acid selectivity by inhibiting acetaldehyde synthesis more strongly than its oxidation to acetic acid, thus minimizing prevalent acetaldehyde concentrations and its intervening conversion to CO(x). Kinetic and isotopic effects indicate that C-H bond activation in chemisorbed ethoxide species limits acetaldehyde synthesis rates and overall rates of ethanol conversion to acetic acid. The VO(x) component in Mo-V-Nb is responsible for the high reactivity of these materials. Mo and Nb oxide components increase the accessibility and reducibility of VO(x) domains, while concurrently decreasing the number of unselective V-O-Ti linkages in VO(x) domains dispersed on TiO(2). PMID- 17912731 TI - Total synthesis and configurational assignment of pasteurestin A and B. PMID- 17912733 TI - Linking two distinct layered networks of nanosized {Ln18} and {Cu24} wheels through isonicotinate ligands. AB - A new series of heterolanthanide(III)-copper(I) wheel-cluster complexes [Ln6(micro3-O)2](IN)18-[Cu8(micro4-I)2(micro2-I)3].H3O (IN=isonicotinate; Ln=Y 1, Nd 2, Dy 3, Gd 4, Sm 5, Eu 6, Tb 7) were prepared by hydrothermal reaction at low pH. X-ray crystallographic studies reveal that two unusual trinuclear [Ln3(micro3 O)] and tetranuclear [Cu4(micro4-I)] cores are successfully used as secondary building units to make two different nanosized wheels [Ln18(micro3-O)6(CO2)48](6 ), {Ln18}, and [Cu24(micro4-I)6(micro2-I)12]6+, {Cu24}, with 12-rings and a diameter of 26.7 and 26.4 A, respectively. The wheels are further assembled into two-dimensional (2D) {Ln18} and {Cu24} networks, the linkages between two distinct layered networks of {Ln18} and {Cu24} wheels by IN pillars along the c axis giving a series of unprecedented three-dimensional (3D) sandwich frameworks. To our knowledge, compounds 1-7 are the first examples containing two different layered networks of nanosized Ln and transition metal (TM) wheels in wheel cluster chemistry. The IR, UV/Vis, thermogravimetric analysis (TGA), luminescent, and magnetic properties of these complexes were also studied. PMID- 17912734 TI - Learning from nature: beta-sheet-mimicking copolymers get organized. AB - The solution structures formed by coil-coil copolymers arise from the selective solvation of one of the two blocks and have been well described. In most cases in such relatively simple synthetic structures there are no specific attractive forces that can aid the aggregation process. Nature, however, provides plenty of inspiring polymeric architectures that are shaped and ordered hierarchically by noncovalent forces. The high level of structural definition displayed by proteins, for example, is unmatched by synthetic polymers. An emerging area of interest in polymer science tries to combine the best of both worlds, the natural and the synthetic, by conjugating synthetic polymers and beta-sheet-forming peptides. Understanding the supramolecular organization of the block copolymers driven exclusively by the intermolecular attractive forces of the peptide sequence is of particular interest. Not only do these peptide-polymer hybrid structures present an interesting new class of materials, they can also provide important insights into self-organization processes prevalent in nature. PMID- 17912735 TI - Encapsulated N-heterocyclic carbenes in silicones without reactivity modification. PMID- 17912736 TI - Functional G-quartet macroscopic membrane films. PMID- 17912737 TI - Cinchona-alkaloid-catalyzed enantioselective direct aldol-type reaction of oxindoles with ethyl trifluoropyruvate. PMID- 17912738 TI - Promising material for infrared nonlinear optics: NaI(3)O(8) salt containing an octaoxotriiodate(V) anion formed from condensation of [IO(3)](-) ions. PMID- 17912739 TI - Neural pathways for the processing of alarm pheromone in the ant brain. AB - Social insects like ants exhibit sophisticated communication by means of pheromones, one example of which is the use of alarm pheromones to alert nestmates for colony defense. In the ant Camponotus obscuripes, we have reported that information about formic acid and n-undecane, alarm pheromone components, is processed in a set of specific glomeruli in the antennal lobe (primary olfactory center). Alarm pheromone signals are then transmitted, mainly via uniglomerular projection neurons (uni-PNs), to the protocerebrum (PR), where sensory signals are integrated to form motor commands for behavioral responses. In this study, we physiologically and morphologically characterized 63 alarm pheromone-sensitive PR neurons in ants by using intracellular recording and staining techniques. Most of the pheromone-sensitive PR neurons had dendrites in the mushroom body (MB), the lateral horn, or the medial PR. Some neurons with dendrites in these areas responded specifically to formic acid or n-undecane and may participate in the control of specific behavioral responses to each pheromone component. Other neurons responded also to non-pheromonal odors, in contrast to uni-PNs, most of which responded specifically to alarm pheromones. Responses to non-pheromonal odors were most prominent in efferent neurons of the MB lobe, suggesting that they may participate in integration of pheromonal and non-pheromonal information. We found a class of PR neurons that receives input in all of these pheromone processing areas and terminates in a variety of premotor areas. These neurons may participate in the control of pheromone-sensitized aggressive behavior, which is triggered by non-pheromonal sensory stimuli associated with a potential enemy. PMID- 17912740 TI - Distribution of corticotropin-releasing hormone in the developing zebrafish brain. AB - Corticotropin-releasing hormone (CRH) plays a central role in the physiological regulation of the hypothalamus-pituitary-adrenal/interrenal axis mediating endocrine, behavioral, autonomic, and immune responses to stress. Despite the wealth of knowledge about the physiological roles of CRH, the genetic mechanisms by which CRH neurons arise during development are poorly understood. As a first step toward analyzing the molecular and genetic pathways involved in CRH lineage specification, we describe the developmental distribution of CRH neurons in the embryonic zebrafish, a model organism for functional genomics and developmental biology. We searched available zebrafish expressed sequence tag (EST) databases for CRH-like sequences and identified one EST that contained the complete zebrafish CRH open reading frame (ORF). The CRH precursor sequence contained a signal peptide, the CRH peptide, and a cryptic peptide with a conserved sequence motif. RT-PCR analysis showed crh expression in a wide range of adult tissues as well as during embryonic and larval stages. By whole-mount in situ hybridization histochemistry, discrete crh-expressing cell clusters were found in different parts of the embryonic zebrafish brain, including telencephalon, preoptic region, hypothalamus, posterior tuberculum, thalamus, epiphysis, midbrain tegmentum, and rostral hindbrain and in the neural retina. The localization of crh mRNA within the preoptic region is consistent with the central role of CRH in the teleost stress response through activation of the hypothalamic-pituitary-interrenal axis. The widespread distribution of CRH-synthesizing cells outside the preoptic region suggests additional functions of CRH in the embryonic zebrafish brain. PMID- 17912742 TI - The potassium channel KCNQ5/Kv7.5 is localized in synaptic endings of auditory brainstem nuclei of the rat. AB - KCNQ, also called Kv7, is a family of voltage-dependent potassium channels with important roles in excitability regulation. Of its five known subunits, KCNQ5/Kv7.5 is extensively expressed in the central nervous system and it contributes to the generation of M-currents. The distribution of KCNQ5 was analyzed in auditory nuclei of the rat brainstem by high-resolution immunocytochemistry. Double labeling with anti-KCNQ5 antibodies and anti synaptophysin or anti-syntaxin, which mark synaptic endings, or anti-microtubule associated protein 2 (MAP2) antibodies, which mark dendrites, were used to analyze the subcellular distribution of KCNQ5 in neurons in the cochlear nucleus, superior olivary complex, nuclei of the lateral lemniscus, and inferior colliculus. An abundance of KCNQ5 labeling in punctate structures throughout auditory brainstem nuclei along with colocalization with such synaptic markers suggests that a preferred localization of KCNQ5 is in synaptic endings in these auditory nuclei. Punctate KCNQ5 immunoreactivity virtually disappeared from the cochlear nucleus after cochlea removal, which strongly supports localization of this channel in excitatory endings of the auditory nerve. Actually, neither glycinergic endings, labeled with an anti-glycine transporter 2 (GlyT2) antibody, nor gamma-aminobutyric acid (GABA)ergic endings, labeled with an anti-glutamic acid decarboxylase (GAD65) antibody, contained KCNQ5 immunoreactivity, suggesting that KCNQ5 is mostly in excitatory endings throughout the auditory brainstem. Overlap of KCNQ5 and MAP2 labeling indicates that KCNQ5 is also targeted to dendritic compartments. These findings predict pre- and postsynaptic roles for KCNQ5 in excitability regulation in auditory brainstem nuclei, at the level of glutamatergic excitatory endings and in dendrites. PMID- 17912741 TI - Chemical and morphological alterations of spines within the hippocampus and entorhinal cortex precede the onset of Alzheimer's disease pathology in double knock-in mice. AB - Mice with knock-in of two mutations that affect beta amyloid processing and levels (2xKI) exhibit impaired spatial memory by 9-12 months of age, together with synaptic plasticity dysfunction in the hippocampus. The goal of this study was to identify changes in the molecular and structural characteristics of synapses that precede and thus could exert constraints upon cellular mechanisms underlying synaptic plasticity. Drebrin A is one protein reported to modulate spine sizes and trafficking of proteins to and from excitatory synapses. Thus, we examined levels of drebrin A within postsynaptic spines in the hippocampus and entorhinal cortex. Our electron microscopic immunocytochemical analyses reveal that, by 6 months, the proportion of hippocampal spines containing drebrin A is reduced and this change is accompanied by an increase in the mean size of spines and decreased density of spines. In the entorhinal cortex of 2xKI brains, we detected no decrement in the proportion of spines labeled for drebrin A and no significant change in spine density at 6 months, but rather a highly significant reduction in the level of drebrin A immunoreactivity within each spine. These changes are unlike those observed for the somatosensory cortex of 2xKI mice, in which synapse density and drebrin A immunoreactivity levels remain unchanged at 6 months and older. These results indicate that brains of 2xKI mice, like those of humans, exhibit regional differences of vulnerability, with the hippocampus exhibiting the first signatures of structural changes that, in turn, may underlie the emergent inability to update spatial memory in later months. PMID- 17912743 TI - A model of early molecular regionalization in the chicken embryonic pretectum. AB - The pretectal region of the brain is visualized as a dorsal region of prosomere 1 in the caudal diencephalon, including derivatives from both the roof and alar plates. Its neuronal derivatives in the adult brain are known as pretectal nuclei. The literature is inconsistent about the precise anteroposterior delimitation of this region and on the number of specific histogenetic domains and subdomains that it contains. We performed a cross-correlated gene-expression map of this brain area in chicken embryos, with the aim of identifying differently fated pretectal domains on the basis of combinatorial gene expression patterns. We examined in detail Pax3, Pax6, Pax7, Tcf4, Meis1, Meis2, Nkx2.2, Lim1, Dmbx1, Dbx1, Six3, FoxP2, Zic1, Ebf1, and Shh mRNA expression, as well as PAX3 and PAX7 immunoreaction, between stages HH11 and HH28. The patterns analyzed serve to fix the cephalic and caudal boundaries of the pretectum and to define three molecularly distinct anteroposterior pretectal domains (precommissural, juxtacommissural, and commissural) and several dorsoventral subdomains. These molecular specification patterns are established step by step between stages HH10 and HH18, largely before neurogenesis begins. This set of gene-architectonic data constitutes a useful scaffold for correlations with fate maps and other experimental embryologic results and may serve as well for inquiries on homologies in this part of the brain. PMID- 17912744 TI - Atypical embryonic synapses fail to regenerate in adulthood. AB - Functional recovery following central nervous system (CNS) injury in adult animals may depend on the reestablishment of the precise pattern of connections made during development. When the nervous system is injured during embryonic development, functional recovery may involve the formation of atypical connections. Can such atypical synapses regenerate in adults, particularly in a nervous system known for its capacity for repair? When the S interneuron in one segmental ganglion of the leech Hirudo is killed during development, two neighboring S cells extend their axons into the ganglion and restore function by making electrical synapses with the usual synaptic targets of the killed S cell. Although adult S-cell axons reliably regenerated their usual synaptic connections, the novel synapses induced following embryonic injury failed to regenerate in adults. In these preparations severed S-cell axons did not reach the denervated ganglion but grew close to it, independent of the distance required to grow. Thus, the developmental changes that permit aberrant but functional connections in embryos do not lead to a similar change in the capacity for axon growth and subsequent synapse regeneration in adults. PMID- 17912745 TI - Effects of inflammation on the ultrastructural localization of spinal cord dorsal horn group I metabotropic glutamate receptors. AB - Inflammatory pain is thought to induce functional plasticity of spinal dorsal horn neurons and may produce changes in glutamate receptor expression. Plasticity of group I metabotropic glutamate receptors (mGluR1 and mGluR5) is important in various neuronal systems, and these receptors are also known to modulate nociceptive neurotransmission in the spinal dorsal horn. The present study aimed at determining whether persistent inflammatory pain produces alterations in intracellular and plasma membrane-associated mGluR1alpha and mGluR5 in spinal cord dorsal horn. Persistent inflammation was induced in male Long Evans rats by a unilateral intraplantar injection of 100 muL of complete Freund's adjuvant (CFA). Three days after the CFA injection thermal withdrawal latencies were obtained prior to processing of transverse spinal cord sections for preembedding immunogold labeling after incubation in primary antibody for mGluR1alpha or mGluR5. Using electron microscopy, we quantified immunogold-labeled mGluR1alpha and mGluR5 profiles, located in lamina V and I-II, respectively, of both CFA treated rats and untreated control rats. Compared to untreated rats, CFA-treated rats had a significant increase in the number of plasma membrane-associated mGluR5 immunogold-labeled particles in lamina I-II neurons of the spinal cord. Although no changes to mGluR1alpha expression were found in CFA-treated rats, plasma membrane-associated mGluR1alpha was significantly closer to the synapse. Therefore, in CFA-treated rats there was a specific increase in the ratio of plasma membrane-associated versus intracellular immunogold-labeled particles for mGluR5, and lateral movement of mGluR1alpha toward the synapse, indicating that peripheral inflammation-induced trafficking of group I mGluRs in spinal dorsal horn neurons may be an important factor in the development of plastic changes associated with inflammation-induced chronic pain. PMID- 17912747 TI - Voluntary activation during knee extensions in severely deconditioned patients with chronic obstructive pulmonary disease: benefit of endurance training. AB - As a consequence of hypoxemia or detraining, a deficit in voluntary activation may contribute to the reduction of strength in chronic obstructive pulmonary disease (COPD). This was studied in 13 severely deconditioned COPD patients and eight healthy subjects. Exercise tolerance, fat-free mass, and quadriceps maximal voluntary contractions (MVC) were evaluated. Magnetic stimulation was used to measure maximal quadriceps twitch tension (TwQp) at rest and during a voluntary contraction (superimposed twitch: TwQs). A maximal recruitment fraction (MRF) was assessed by the ratio of the true MVC to the maximal achievable strength (using regression of TwQs vs. volitional strength at four submaximal intensities). MRF was significantly lower in COPD than in controls (89+/-20 vs. 109+/-6%, P<0.01). Seven COPD patients completed 3-month endurance training. After rehabilitation, patients had greater MVC (+24+/-26 N), associated with a slightly greater MRF (+10+/-11%). Changes in MRF were independently associated with changes in MVC (r=0.85, P=0.03). Deficit in voluntary activation of quadriceps muscle may therefore contribute to muscle weakness in severe COPD patients and is reversed by training. PMID- 17912746 TI - Spatial and temporal activation of brain regions in hibernation: c-fos expression during the hibernation bout in thirteen-lined ground squirrel. AB - Hibernation results in dramatic changes in body temperature and metabolism; however, the central nervous system remains active during deep torpor. By cloning c-fos cDNA from the 13-lined ground squirrel (Spermophilus tridecemlineatus) and using squirrel c-fos mRNA probe for in situ hybridization histochemistry, we systematically analyzed and identified specific brain regions that were activated during six different phases of the hibernation bout. During entrance into torpor, we detected activation of the ventrolateral subdivision of the medial preoptic area ('thermoregulatory center'), and the reticular thalamic nucleus, which is known to inhibit the somatomotor cortex. During torpor, c-fos expression in the cortex was suppressed while the reticular thalamic nucleus remained uniformly active. Throughout torpor the suprachiasmatic nucleus ('biological clock') showed increasing activity, likely participating in phase-change regulation of the hibernation bout. Interestingly, during torpor very strong c-fos activation was seen in the epithelial cells of the choroid plexus and in tanycytes at the third ventricle, both peaking near the beginning of arousal. In arousal, activity of the suprachiasmatic and reticular thalamic nuclei and choroid epithelial cells diminished, while ependymal cells in the lateral and fourth ventricles showed stronger activity. Increasing body temperature during arousal was driven by the activation of neurons in the medial part of the preoptic area. In interbout awake animals, we demonstrated the activation of hypothalamic neurons located in the arcuate nucleus and the dorsolateral hypothalamus, areas involved in food intake. Our observations indicate that the hibernation bout is closely regulated and orchestrated by specific regions of the central nervous system. J. Comp. Neurol. 505:443-458, 2007. (c) 2007 Wiley-Liss, Inc. PMID- 17912748 TI - Effect of innervation-zone distribution on estimates of average muscle-fiber conduction velocity. AB - Muscle-fiber conduction velocity (MFCV) may be a diagnostic variable, but the factors affecting its estimate from surface electromyographic (SEMG) signals are still not fully understood. We investigated the effect of innervation-zone distribution on estimates of average MFCV from simulated SEMG signals. Single fiber action potentials were simulated using a computer model that describes an idealized cylindrical volume conductor comprised of bone, muscle, subcutaneous, and skin tissues. A model describing the characteristics and control of a motor neuron pool was used to simulate 60 s of SEMG. Five innervation-zone distributions, inclined with respect to the fiber direction, were simulated within the muscle tissue, based on data from the literature and an experimental analysis. Two parameters were varied concurrently: (1) excitation level (5%-100%, 5% increments); and (2) subcutaneous tissue thickness (4 and 10 mm). The MFCV estimates were affected by the distribution of innervation zones with bias varying with excitation level and thickness of subcutaneous tissue. The MFCV estimates ranged from 3.95 to 11.67 m/s for single-differential and from 3.99 to 5.23 m/s for double-differential recordings when all simulated motor units were assigned the same conduction velocity of 4 m/s. Amplitude and characteristic spectral frequencies were also influenced by the distribution of innervation zones. The effect of innervation-zone distribution on MFCV estimates is thus substantial and may be a significant confounding factor in experimental and clinical studies on muscles with diffuse innervation zones. PMID- 17912749 TI - Effects of CRF2R agonist on tumor growth and cachexia in mice implanted with Lewis lung carcinoma cells. AB - Previous studies have demonstrated an effect of corticotropin-releasing factor 2 receptor (CRF2R) agonists in the maintenance of skeletal muscle mass. The aim of this study was to evaluate the effects of a CRF2R agonist in preserving skeletal muscle in a mouse cachexia model. Implantation of a fast-growing tumor to mice (Lewis lung carcinoma) resulted in a clear cachectic state characterized by a profound muscle wasting. We found that administration of a CRF2R agonist (PG 873637) at 100 microg/kg/day by means of osmotic minipumps to tumor-bearing mice resulted in beneficial effects on muscle weight loss. Thus, muscle loss was partially reversed by the CRF2R agonist at different stages of tumor growth (at day 14 after tumor inoculation: 12% in tibialis posterior; 9% in gastrocnemius; and 48% in soleus). Moreover, the CRF2R agonist significantly reduced both the number of metastases and their mass (at day 19 after tumor inoculation: 66% and 61%, respectively). These data suggest a potentially beneficial effect of the CRF2R agonist in preserving skeletal muscle during cancer cachexia and open a line of research for the development of new therapeutic approaches for the treatment of muscle wasting associated with cancer. PMID- 17912750 TI - Myogenesis in human denervated muscle biopsies. AB - Electron microscopic studies of long-term denervated rat muscles have identified very small, immature myofibers that are believed to arise from detached satellite cells that have fused to form new fibers within the interstitial space. At present, it is unknown whether and to what extent equivalent fibers exist in denervated human muscle. Serial sections of muscle biopsies from 66 patients diagnosed with polyneuropathy or amyotrophic lateral sclerosis were immunolabeled with anti-NCAM and anti-neonatal myosin heavy chain monoclonal antibodies that are both neurally and developmentally regulated. We evaluated 200 myofibers in each section. Of the biopsy specimens, 75% contained small myofibers that showed a thin perinuclear cytoplasmic rim. Small fibers expressing neonatal myosin heavy chain (MHCn+) were found in all of these biopsies (100%) and NCAM+ fibers in 98%. The percentage of MHCn+ small fibers averaged 82% and NCAM+ small myofibers averaged 40%. The percentage of NCAM+ small fibers was significantly lower than that of MHCn+ fibers. In contrast, the percentage of MHCn+ vs. NCAM+ angular atrophic fibers did not show a significant difference. A substantial subset of neurogenic biopsies showed small fibers that differ from angular atrophic fibers both in size and expression pattern of MHCn and NCAM. Myogenesis appears to be a frequent finding in neurogenic atrophy. PMID- 17912751 TI - Reliability of a novel neurostimulation method to study involuntary muscle phenomena. AB - Experimental methods involving painful electrical stimulation of a peripheral nerve showed the existence of a minimum stimulation frequency capable of inducing cramp, termed "threshold frequency" (TF). Our aim was to test an alternative method to induce fasciculations and cramps electrically. Two daily sessions of electrical stimulation of the abductor hallucis muscle were performed in 19 volunteers on 3 days: stimulation trains of 150 monophasic square pulses (duration 152 micros) of increasing frequency (current intensity 30% higher than maximal; frequency of the first trial, 4 pps; recovery between trials, 1 min) were delivered to the main muscle motor point until a cramp developed. Once a cramp was induced the protocol was repeated after 30 min. To verify by electromyography that cramp occurred, a surface electrode array was placed between the motor point and the distal tendon. Ambient and skin temperature were kept constant in all sessions. Fasciculations and cramps were elicited in all subjects. We observed the following median (interquartile range) values of TF: day 1 (session 1), 13 (6) pps; day 1 (session 2), 16 (4) pps; day 2 (session 1), 16 (6) pps; day 2 (session 2), 18 (6) pps; day 3 (session 1), 17 (4) pps; day 3 (session 2), 18 (8) pps. TF intersession intraclass correlation coefficients were 0.82, 0.92, and 0.90 for days 1, 2, and 3, respectively. TF interday intraclass correlation coefficient was 0.85. The absence of pain due to the stimulation and the demonstration of TF reliability support the use of our method for the study of involuntary muscle phenomena. PMID- 17912752 TI - Novel mutation in KCNA1 causes episodic ataxia with paroxysmal dyspnea. AB - Episodic ataxia type 1 (EA1) is an autosomal-dominant neurological disease caused by point mutations in the potassium channel-encoding gene KCNA1. It is characterized by attacks of ataxia and continuous myokymia. Respiratory muscle involvement has not been previously reported in EA1. We clinically evaluated a family with features of EA1 and paroxysmal shortness of breath. Coding and flanking intronic regions of KCNA1 were sequenced. We identified a novel 3 nucleotide deletion mutation in KCNA1 in the affected individuals. Our findings of a deletion mutation with unusual respiratory muscle involvement expand the genetic and clinical spectrum of EA1. PMID- 17912753 TI - Ulnar sensory nerve impairment at the wrist in carpal tunnel syndrome. AB - In previous studies, changes in impulse transmission of ulnar motor axons have been documented in patients with carpal tunnel syndrome (CTS). We examined ulnar sensory conduction in 144 CTS hands. In particular, conduction parameters of the dorsal ulnar cutaneous branch (DUC) running outside Guyon's canal were compared with those of the superficial sensory branches (U4 and U5) passing through the canal. U4 and U5 response amplitudes and U5 conduction velocity were significantly lower than in controls. Conduction parameters of the DUC were similar in both groups. Patients with more severely impaired median conduction had smaller ulnar sensory action potentials. We propose that the ulnar nerve may be subject to compression in Guyon's canal as a consequence of high pressure in the carpal tunnel of CTS patients. This may provide insights into the mechanisms underlying extra-median spread of sensory symptoms in CTS patients. PMID- 17912754 TI - From the Mediterranean coast to the shores of Lake Ontario: CAPRI's premiere on the American continent. PMID- 17912755 TI - Differential native state ruggedness of the two Ca2+-binding domains in a Ca2+ sensor protein. AB - Characterization of near-native excited states of a protein provides insights into various biological functions such as co-operativity, protein-ligand, and protein-protein interactions. In the present study, we investigated the ruggedness of the native state of EhCaBP using nonlinear temperature dependence of backbone amide-proton chemical shifts. EhCaBP is a two-domain EF-hand calcium sensor protein consisting of two EF-hands in each domain and binds four Ca2+ ions. It has been observed that approximately 30% of the residues in the protein access alternative conformations. Theoretical modeling suggested that these low energy excited states are within 2-3 kcal/mol from the native state. Further, it is interesting to note that the residues accessing alternative conformations are more dominated in the C-terminal domain compared with its N-terminal counterpart suggesting that the former is more rugged in its native state. These distinct characteristics of N- and C-terminal domains of a calcium sensor protein belonging to the super family of calmodulin would have implications for domain dependent Ca2+ signaling pathways. PMID- 17912756 TI - Screening of the active site from water by the incoming ligand triggers catalysis and inhibition in serine proteases. AB - The pKa of the catalytic His57 N(epsilon)H in the tetrahedral complex (TC) of chymotrypsin with trifluoromethyl ketone inhibitors is 4-5 units higher relative to the free enzyme (FE). Such stable TC's, formed with transition state (TS) analog inhibitors, are topologically similar to the catalytic TS. Thus, analysis of this pKa shift may shed light on the role of water solvation in the general base catalysis by histidine. We applied our QM/SCRF(VS) approach to study this shift. The method enables explicit quantum mechanical DFT calculations of large molecular clusters that simulate chemical reactions at the active site (AS) of water solvated enzymes. We derived an analytical expression for the pKa dependence on the degree of water exposure of the ionizable group, and on the total charge in the enzyme AS, Q(A) and Q(B), when the target ionizable functional group (His57 in this study) is in the acidic (A) and basic (B) forms, respectively. Q2(B) > Q2(A) both in the FE and in the TC of chymotrypsin. Therefore, water solvation decreases the relative stability of the protonated histidine in both. Ligand binding reduces the degree of water solvation of the imidazole ring, and consequently elevates the histidine pKa. Thus, the binding of the ligand plays a triggering role that switches on the cascade of catalytic reactions in serine proteases. PMID- 17912757 TI - Mn/Fe superoxide dismutase interaction fingerprints and prediction of oligomerization and metal cofactor from sequence. AB - Fe- and Mn-containing superoxide dismutase (sod) enzymes are closely related and similar in both amino acid sequence and structure, but differ in their mode of oligomerization and in their specificity for the Fe or Mn cofactor. The goal of the present work is to identify and analyze the sequence and structure characteristics that ensure the cofactor specificities and the oligomerization modes. For that purpose, 374 sod sequences and 17 sod crystal structures were collected and aligned. These alignments were searched for residues and inter residue interactions that are conserved within the whole sod family, or alternatively, that are specific to a given sod subfamily sharing common characteristics. This led us to define key residues and inter-residue interaction fingerprints in each subfamily. The comparison of these fingerprints allows, on a rational basis, the design of mutants likely to modulate the activity and/or specificity of the target sod, in good agreement with the available experimental results on known mutants. The key residues and interaction fingerprints are furthermore used to predict if a novel sequence corresponds to a sod enzyme, and if so, what type of sod it is. The predictions of this fingerprint method reach much higher scores and present much more discriminative power than the commonly used method that uses pairwise sequence comparisons. PMID- 17912758 TI - The HhH domain of the human DNA repair protein XPF forms stable homodimers. AB - The human XPF-ERCC1 protein complex plays an essential role in nucleotide excision repair by catalysing positioned nicking of a DNA strand at the 5' side of the damage. We have recently solved the structure of the heterodimeric complex of the C-terminal domains of XPF and ERCC1 (Tripsianes et al., Structure 2005;13:1849-1858). We found that this complex comprises a pseudo twofold symmetry axis and that the helix-hairpin-helix motif of ERCC1 is required for DNA binding, whereas the corresponding domain of XPF is functioning as a scaffold for complex formation with ERCC1. Despite the functional importance of heterodimerization, the C-terminal domain of XPF can also form homodimers in vitro. We here compare the stabilities of homodimeric and heterodimeric complexes of the C-terminal domains of XPF and ERCC1. The higher stability of the XPF HhH complexes under various experimental conditions, determined using CD and NMR spectroscopy and mass spectrometry, is well explained by the structural differences that exist between the HhH domains of the two complexes. The XPF HhH homodimer has a larger interaction interface, aromatic stacking interactions, and additional hydrogen bond contacts as compared to the XPF/ERCC1 HhH complex, which accounts for its higher stability. PMID- 17912759 TI - Intra-articular electrotransfer of plasmid encoding soluble TNF receptor variants in normal and arthritic mice. AB - BACKGROUND: Anti-inflammatory gene therapy is promising in inflammatory diseases such as rheumatoid arthritis (RA). We have previously demonstrated that intra muscular (i.m.) electrotransfer (ET) of plasmids encoding three different human tumor necrosis factor-alpha-soluble receptor I variants (hTNFR-Is) exert protective effects in an experimental RA model. However, such a systemic approach could be responsible for side effects. The present study aimed at performing an intra-articular (i.a.) gene therapy by electrotransfer using the hTNFR-Is plasmids. METHODS AND RESULTS: We evaluated targeting of mice joints by CCD optical imaging after i.a. ET of a luciferase-encoding plasmid and we showed that ET led to strongly increased transgene expression in a plasmid dose-dependent manner. Moreover, articular and seric hTNFR-Is was detectable for 2 weeks. As expected, systemic hTNFR-Is rates were lower after i.a. ET than after i.m. ET. A longer protein secretion could be achieved with several i.a. ETs. Also, we observed that hTNFR-Is expression within arthritic joints was slightly higher than in normal joints. CONCLUSIONS: In collagen-induced arthritis (CIA), a mouse model for RA, we demonstrated that hTNFR-Is/mIgG1-encoding plasmid i.a. ET decreased joint destruction in the ankles. In conclusion, our results suggest that local TNFR-Is gene therapy may play a role in decreasing joint destruction in CIA. PMID- 17912760 TI - Allergy shots or automobiles: which is more dangerous? PMID- 17912761 TI - Case report: Patient adherence to drug regimens vital to treatment. PMID- 17912762 TI - Deborah Charlesworth. PMID- 17912763 TI - A pipeline for Europe. PMID- 17912764 TI - Proceedings of the 8th International Symposium on Drug Analysis. May 16-19, 2006. Namur, Belgium. PMID- 17912765 TI - Music from the heart--in praise of auscultation. Interview by Keith Barnard. PMID- 17912766 TI - The founder of a key European association. Interview by Ingrid Torjesen. PMID- 17912767 TI - Uncovering the dark side of TNF. AB - In the early 1980s, in search of the cytokine that triggers disease- associated weight loss and lethal shock, Anthony Cerami and Bruce Beutler purified "cachectin." They soon found out that their malevolent cytokine already had another name-tumor necrosis factor (TNF)-and was being lauded as a potential antitumor agent. PMID- 17912769 TI - Doctors to pay for patients' medicine in Germany. PMID- 17912768 TI - Current awareness in phytochemical analysis. PMID- 17912770 TI - Community workers key to improving Africa's primary care. PMID- 17912771 TI - Current awareness on yeast. PMID- 17912772 TI - Abstracts of the 39th Annual Congress of the International Society of Paediatric Oncology (SIOP), November 1-3, 2007, Mumbai, India. PMID- 17912773 TI - Abstracts of the 62nd Annual Session of the Surgical Forum 2007 Clinical Congress, October 7-11, 2007, New Orleans, Louisiana, USA. PMID- 17912774 TI - Headache drug lacks evidence. PMID- 17912775 TI - High-Speed, swept-source optical coherence tomography: a 3-dimensional view of anterior chamber angle recession. PMID- 17912776 TI - Interprofessional wound caring and the IWC Model. PMID- 17912777 TI - An alternative solution for difficult-to-manage colostomies in the descending and sigmoid colon. PMID- 17912779 TI - How to catch more cases of celiac disease. PMID- 17912778 TI - Giving the gift of time. Interview by Barbara Zeiger. PMID- 17912780 TI - Tricyclics, capsaicin, and older anticonvulsants are best for neuropathy. PMID- 17912781 TI - Public initiatives to improve the prevention, detection, and management of hypertension. PMID- 17912782 TI - What are the best agents for preventing VTE in hospital patients? PMID- 17912783 TI - Do glucose monitors help type 2 patients with long-term control? PMID- 17912784 TI - Designation of oripavine as a basic class of controlled substance. Final rule. AB - This is a final rule issued by the Drug Enforcement Administration (DEA) designating oripavine (3-O-demethylthebaine or 6,7,8,14-tetradehydro-4,5-alpha epoxy-6-methoxy-17-methylmorphinan-3-ol) as a basic class in schedule II of the Controlled Substances Act (CSA). Although oripavine was not previously listed in schedule II of the CSA, it has been controlled in the United States as a derivative of thebaine and, as such, is controlled as a schedule II controlled substance which includes "Opium and opiate, and any salt, compound, derivative, or preparation of opium or opiate." Oripavine is a derivative of thebaine, a natural constituent of opium, hence oripavine has been and continues to be, by virtue of the definition of "narcotic drug", a schedule II controlled substance. International control of oripavine in schedule I of the 1961 Single Convention on Narcotic Drugs (1961 Convention) during the 50th session of the Commission on Narcotic Drugs (CND) in 2007 prompted the DEA to specifically designate oripavine as a basic class of controlled substance in schedule II of the CSA. PMID- 17912785 TI - [Abstracts of the International Symposium on Malaria, 16-19 July 2007, Caracas, Venezuela]. PMID- 17912786 TI - TRICARE; TRICARE retiree dental program (TRDP) basic benefit descriptions and administrative corrections. Final rule. AB - This final rule amends TRICARE Retiree Dental Program (TRDP) Basic benefit descriptions by replacing specific American Dental Association (ADA) dental procedure codes and nomenclature with general benefit categories and descriptions. This revision is necessary to keep the regulation current, since dental procedure codes are added, revised, and deleted on a regular basis. This final rule does not change or eliminate any benefits that are currently available under the TRDP program. This final rule also revises several incorrect, obsolete, or historical terms pertaining to the TRICARE program, and removes an inaccurate statement regarding appeals and grievances. PMID- 17912788 TI - Abstracts of the 1st MC-GARD (Marie Curie-Genome Architecture in Relation to Disease) Meeting, 3-5 May 2007, Amsterdam, The Netherlands. PMID- 17912787 TI - TRICARE; Changes included in the John Warner National Defense Authorization Act for Fiscal Year 2007; authorization of anesthesia and other costs for dental care for children and certain other patients. Final rule. AB - This final rule implements section 702 of the John Warner National Defense Authorization Act for Fiscal Year 2007, Public Law 109-364. The rule provides coverage of contracted medical care with respect to dental care beyond that care required as a necessary adjunct to medical or surgical treatment. The entitlement of institutional and anesthesia services is authorized in conjunction with non covered dental treatment for patients with developmental, mental, or physical disabilities or for pediatric patients age 5 or under. This final rule does not eliminate any contracted medical care that is currently covered for spouses and children. The entitlement of anesthesia services includes general anesthesia services only. Institutional services include institutional benefits associated with both hospital and in-out surgery settings. Patients with developmental, mental, or physical disabilities are those patients with conditions that prohibit dental treatment in a safe and effective manner. Therefore, it is medically or psychologically necessary for these patients to require general anesthesia for dental treatment. PMID- 17912790 TI - Abstracts of the 15th International Congress of the Hungarian Society for Microbiology, July 18-20, 2007, Budapest, Hungary. PMID- 17912789 TI - [Delinquency, crime, and feelings of insecurity: reflections on the case of Mexico]. PMID- 17912791 TI - Culinary innovation, love, and the social organization of learning in a West African city. PMID- 17912792 TI - [Acid pump inhibitor before before gastroscopy for upper gastrointestinal bleeding]. PMID- 17912793 TI - [Kittelbogen medicin.dk]. PMID- 17912795 TI - Bibliography. Current world literature. Oculoplastic and orbital surgery. PMID- 17912794 TI - Pharmacogenomics and warfarin therapy. AB - (1) Dosing algorithms tailored to individual genetic, demographic, and clinical factors may minimize the risk for bleeding during the initiation of warfarin therapy. (2) Pharmacogenomic testing should be used in addition to (rather than replacing) routine International Normalized Ratio (INR) monitoring. (3) Prospective studies are needed to determine whether pharmacogenomic testing improves patient outcomes, identify which subgroups of patients may benefit, and clarify the risks and costs associated with the use of these tests. Several randomized controlled trials are currently evaluating the impact of pharmacogenomics on dosing accuracy, time to achieve and maintain target INR, incidence of bleeding or thromboembolic events, and monitoring requirements. (4) In August 2007, the FDA updated the product label for warfarin to include genetic variations in CYP2C9 and VKORC1 as one of the factors to consider for more precise initial dosing. Guidelines for pharmacogenomics-based warfarin dosing are under development. PMID- 17912796 TI - Have some calcium with your chocolate. PMID- 17912798 TI - Social Security: when to start collecting. You can take benefits as early as age 62, but odds are you'll get more money overall if you can wait. PMID- 17912797 TI - Sweeteners: how the brands measure up. PMID- 17912799 TI - HPV vaccine: beyond the hype. PMID- 17912800 TI - [Cattle babesiosis]. AB - Babesia parasites are intraerytrocytic Protozoa that infect wide range of domestic and wild animals and occasionally man causing babesiosis (piroplasmosis). Babesiosis also known in cattle as tick fever or red water fever is most important arthropod-borne disease of bovinae ungulates worldwide with areas of Africa, Asia, South and Central America, Australia and finally Europe. Since Smith and Kilborne had first described potential role of ticks in spread of piroplasmosis within animals, only United States till end of 50 of XX century eradicated the disease from the continent. In other, especially African countries, the problem seems to be of great economic importance. In this review all species of Babesia known to date to be infective to cattle are described with emphasis on geographical distribution of piroplasmosis, tick vector and pathogenicity of particular species and strains. PMID- 17912801 TI - [Health risks connected with the avian flu virus]. AB - The first human cases of infections with entire avian influenza strain were caused by subtype A/H5N1/ and occurred in 1997 in Hong Kong. Since this year until 27 March 2007 World Health Organization confirmed over 370 human cases of infections with avian influenza and almost 160 persons died due to this pathogen. At present there is known that person to person transmission of avian influenza virus is limited. Nevertheless, it is possible that with time virus will adapt to human sufficiently to be able to cause next influenza pandemic. Despite intensive studies and global influenza surveillance there is no possible to predict when pandemic will begin and what kind of virus will cause this outbreak. PMID- 17912802 TI - [State of knowledge of helminth fauna of freshwater fishes of Poland]. AB - A total 89 fish and lamprey species has been recorded from Polish freshwater habitats. Twenty-seven of them (30.3%) have not been surveyed for parasitic helminthes. Some of the latter fishes are either rare or not easily accessible. Other live only in specific habitats in scattered localities. An important obstacle for studying parasite faunas of some fishes may be their status on an endangered species. Among the non-surveyed fishes, are those which have been relatively recently introduced to Poland or migrated there on their own. The present paper attempts to review all hitherto not studied helminthologically fish species, their habitats, localities and current protection status. PMID- 17912803 TI - Hydrolases of Hysterothylacium aduncum (Nematoda). AB - BACKGROUND: Enzymatic activity is an indicator of an organism's metabolic rate which depends on, i.e., environmental conditions, developmental stage, physiological stage, and sex. The API ZYM test was applied to compare activities of 19 hydrolases of female and male Hysterothylacium aduncum. MATERIAL AND METHODS: Sexually mature nematodes were isolated from eelpout individuals caught in the Gulf of Gdansk. Enzymatic activity of the hydrolases and the protein content was determined in nematode extracts using API ZYM and Bradford's method, respectively. RESULTS: The females and males tested showed a total of 13 enzymes to be active. The males showed additionally the presence of alpha-fucosidase. Acidic and alkaline phosphatases had very high activities in both sexes; short chain fatty acid esterases, leucine and valine aminopeptidases, alpha glucosidase, and N-acetylglucosaminidase were highly active. H. aduncum showed no trypsin- and chymotrypsin-specific activities; similarly, no activity of alpha galactosidase, alpha-mannosidase, and beta-glucuronidase was revealed. Except for lipase (C14), hydrolases were more active in females than in males, which is related to metabolic rate being higher in females due to their reproductive function. CONCLUSION: Comparison of the results obtained with earlier data produced with API ZYM allowed suggesting that the hydrolase pattern may be more affected by habitat in the host than by the taxonomic affiliation of nematode. PMID- 17912804 TI - [Effect in vitro of albendazole on the kinetics of cytosolic glutathione transferase from the rat liver]. AB - INTRODUCTION: Since the idea of multifunctional mode of action of anthelmintics is considered and in experimental trichinellosis in vivo albendazole seems to act as an allosteric activator of cytosolic GST from mice muscles, in this study a termosensitivity after in vitro incubation with albendazole of purified commercial cytosolic glutathione transferase (GST) from the rat liver was investigated. METHODS: Two extremal temperatures: -80 degrees C and +30 degrees C were used to destroy the dimer in quaternary structure of this enzyme. RESULTS: In control preparations both extremal temperatures destroy this structure, so the Michaelis-Menten kinetic curves of substrate saturation show the typical hyperbolic shape. After a long (15 h) freezing at -80 degrees C or heating (up to 14 h at +30 degrees C) the kinetics of substrate saturation of GST after incubation with albendazole show the sigmoidal or "double sigmoidal" shape, pointing out the quaternary GST structure as a complex of "frozen subunits". Drug inhibits about 6-times the total activity of GST after incubation at +30 degrees C. We conclude that albendazole in vitro influences the structure of cytosolic GST from the rat liver and inhibits its activity, but, in opposite to in vivo study in mouse muscles infected with Trichinella spiralis larvae, does not act as an activator of this enzyme. PMID- 17912805 TI - Wild game as a reservoir of Anaplasma phagocytophilum in north-western Poland. AB - BACKGROUND: The aim of the study was to determine the role of game animals as reservoirs of Anaplasma phagocytophilum, a bacteria species transmitted by Ixodes ricinus ticks, from north-western Poland (Zachodniopomorskie vovoidship). The area under question is endemic for A. phagocytophilum. MATERIAL AND METHODS: Blood and spleen samples were taken from 72 roe deer between April and December 2003. Animals culled during winter did not harbour ticks, on the other hand 155 individuals of Ixodes ricinus were collected from 35 of 43 animals taken during spring. We tested all samples for A. phagocytophilum by PCR amplification of the msp2 gene. An individual was considered infected if pathogens were detected in at least one isolate (blood or a tissue sample). RESULTS: DNA from A. phagocytophilum was not found in isolates from ticks collected from the animals. The general level of infection for the roe deer was 31.94% (23/72). DNA of A. phagocytophilum was most commonly detected in blood samples; only in three cases was anaplasma DNA detected in spleen and not in blood. Ruminants seem to be the most competent reservoir for A. phagocytophilum in north-western Poland. PMID- 17912806 TI - [Potentially pathogenic fungi in the waters of the Charzykowskie Lake in Zaborski Landscape Park]. AB - The occurrence of potentially pathogenic fungal strains in the Charzykowskie Lake and runnels flowing into and out of it was investigated. The study material was obtained in 2005 and in 2006, in the periods of spring intermix and summer stagnation, and in 2005 in the period of autumn intermix. The fungi found in the Charzykowski Lake belonged to 5 genera: Rhodotorula (R. minuta, R. rubra and R. glutinis), Cryptoccocus (C. neoformans, C. laurentii, C. terreus and C. laurentii), Candida (C. inconspicua, C. lusitaniae, C. tropicalis, C. pelliculosa, C. kefir, C. glabrata, C. inconspicua, C. parapsilosis, C. ciferrii and C. colliculosa), Trichosporon (T. cutaneum) and Klockera (K. apiculata). The fungi found in runnels flowing into and out of the Charzykowskie Lake belonged to 4 genera: Rhodotorula (R. rubra and R. glutinis), Cryptoccocus (C. laurentii, C. neoformans, C. albidus and C. terreus), Candida (C. colliculosa, C. lusitaniae, C. tropicalis, C. pelliculosa, C. cifferii, C. glabrata) and Trichosporon (T. cutaneum). PMID- 17912807 TI - [Intestinal digeneans of birds (superfamily Diplostomoidea) of the masurian lakes]. AB - BACKGROUND: The aim of this study was to complete the information on digenean fauna of birds on the basis of the collection of diplostomid digeneans gathered in 1994-1970 during the field investigations directed by prof. Wisniewski in the region of Masurian lakeland. MATERIAL AND METHODS: A total number of 742 birds from the lakes: Goldapiwo, Mamry Polnocne, Swiecajty and Dargin, belonging to 27 species (8 families) were examined. RESULTS AND DISCUSSION: 335 specimens (45.1%) of infected bird hosted 30 species of intestinal digeneans belonging to the family Diplostomidae (Codonocephalus urniger, Conodiplostomum spathula, Diplostomum baeri, D. commutatum, D. pseudospathaceum, D. spathaceum, Hysteromorpha triloba, Neodiplostomum spathoides, Posthodiplostomum brevicaudatum, P. cuticola, Posthodiplostomum sp., Tylodelphys clavata, T. excavata, T. podicipina), Cyathocotylidae (Cyathocotyle prussica, Holostephanus dubinini, Paracoenogonimus ovatus) and Strigeidae (Apatemon gracilis, Apharyngostrigea cornu, Australapatemon minor, Cotylurus brevis, C. cornutus, C. hebraicus, C. raabei, Ichthyocotylurus erraticus, I. pileatus, I. platycephalus, Ophiosoma patagiatum, Parastrigea flexilis, Strigea falconis). Their specificity and frequency (prevalence, mean intensity and abundance), as well as domination status of these species are also discussed. Cotylurus brevis Dubois et Rausch, 1950 is recorded for the first time in Poland. PMID- 17912808 TI - [Intestinal parasites of parrots]. AB - BACKGROUND: The aim of this study was to determine a parasitic species composition, prevalence and intensity of infection in selected parrots. MATERIAL AND METHODS: The studies were carried out on faecal samples of budgerigars Melopsittacus undulatus (n = 36), cockatiels Nymphicus hollandicus (n = 21), grey parrots Psittacus erithacus (n = 18), eastern rosella Platycercus eximius (n = 10) and senegal parrots Poicephalus senegalus (n = 10) using the Willis-Schlaff and McMaster's methods. RESULTS: Protozoans (Isosporidae and Eimeriidae) and nematodes (Ascarididae, Capillaridae and Heterakidae) were detected in the tested samples. Coccidian oocysts were detected in all examined parrots. Isospora and Eimeria oocysts were found in 52.9% Melopsittacus undulatus, 60% Poicephalus senegalus and 66.7% Psittacus erithacus. Mean number of oocysts per gram of feces (OPG) was high: from about 270 to 1500 depending on both parasite and host species. Three species of parrots were infected with the nematodes (Nymphicus hollandicus and Poicephalus senegalus were free from this infection), but only Ascaridia platycerci was present in these hosts, with the highest prevalence (20%) and highest mean number of eggs per gram of feces (EPG = 1242) in Platycercus eximinus. Heterakis gallinarum was observed only in Psittacus erithacus; 16.6% individuals were infected, and EPG was 212. The highest intensity of infection with nematodes of Ascarididae and Capillaridae was in Platycercus eximius. EPG in this species of parrots was 1242 and 2480, respectively. The obtained results show that introduction of parasitological prophylaxis programs is necessary, especially in the larger birds' farmings and zoological shops. PMID- 17912809 TI - [Occurence of the common tick Ixodes ricinus L. in environments of various degree and character of anthropogenic impact]. AB - MATERIAL AND METHODS: The study of the occurence and relative numbers of the common tick Ixodes ricinus in environments of various degree and character of anthropogenic impact was conducted in 3 forest complexes situated in the Lubelskie voivodeship: the Gulowskie Forests (district of Lukow), the Kozlowieckie Forests (district of Lubartow) and the Dabrowa Forest (within the municipal boundaries of Lublin). The adopted method was flagging in the period of peak activity of ticks in the springs of 2005-2006. 7 environments were distinguished, in which a total of 48 censuses were conducted. Relative density of ticks was assessed based on the number of specimens caught by one person in 60 minutes. RESULTS: In total, 2081 specimens were caught; on average 43.4 in a single sample. For comparisons of the size of the population of the parasites in the distinguished environments, the so-called "risk factor" (Z) was formulated. It was calculated by assigning the value of 1.00 to the average number of ticks caught during one census. Ticks were found in all the investigated environments. Their highest average numbers (Z = 1.91) were found in forest tracks, the lowest ones--along the shoreline of water reservoirs. A high density (Z = 1.00) was recorded in an environment of homogenous vegetation structure, defined as the "forest interior". PMID- 17912810 TI - The first record of Aspidogaster limacoides Diesing, 1834 (Aspidogastridae: Aspidogastrea) in Poland. AB - Aspidogaster limacoides Diesing, 1834 was identified in two specimens of roach (Rutilus rutilus L.) from middle Odra River (Lower Silesia, SW Poland). As the species is new to the Polish parasitofauna, a description, measurements and figure are presented. PMID- 17912811 TI - The new locality of Argas reflexus Fabricius, 1794 in Warsaw, Poland. AB - The new locality of Argas reflexus in Warsaw, Poland is recorded. The ticks occur in the lof of apartment blocks, inhabited by pigeons. When the birds were removed by cleaning services, the ticks began to look for a new host and penetrate the flats. Some of the lodgers had been bitten by tick larvae and nymphs. PMID- 17912812 TI - [Morphological and biochemical features of fungi isolated from patients with renal failure]. AB - Patients with renal failure are more frequently at risk of fungal infections than the healthy individuals. The aim of the study was: (1) Evaluation of the prevalence of fungi in biological materials obtained from different ontocenoses from patients with end-stage and chronic renal failure undergoing haemodialysis and conservative treatment, respectively. (2) Species determining of isolated fungal strains and evaluation their morphological and biochemical features with regard to biotyping. (3) Examining the connection between intraspecies features of fungal strains isolated from different ontocenoses of the same patient. The study group comprised 136 persons, including 56 patients with end-stage renal failure dialysed for the mean period of 36.2 (+/- 1.62) months--all patients were on chronic haemodialysis therapy (4 hours sessions, 3 times per week), 50 patients with chronic renal failure undergoing conservative treatment and 30 persons with the negative history of any renal disease--control group. At the moment of the evaluation and collection of samples all patients were in good condition, none of the patients revealed symptoms suggesting possible fungal infection. Material for mycological examinations included washings from the oral cavity and samples of urine and faeces. In order to evaluate morphological and biochemical features of fungi the following methods were applied: (1) macrocultures in solid Sabouraud medium, (2) direct microscopic slides, (3) API 20 C AUX test and (4) API ZYM test (bioMerieux). Totally 385 samples for mycological examinations were collected from different ontocenoses, from which 161 fungal strains were isolated and classified to 17 species from 5 genera. Most strains belonged to the genus Candida 96.9% and the most frequently occurring species was C. albicans (60.3%). Other species from this genus composed 39.7%, among which in 11.8% of cases C. parapsilosis was determined, and the following were: C. guilliermondii and C. humicola--both species isolated in 5.59%, C. glabrata (4.35%), C. rugosa and C. tropicalis (both species were isolated in 2.48%). The remaining isolated fungal strains (3.11 +/- 1.37%) belonged to genera Cryptococcus, Geotrichum, Saccharomyces and Trichosporon. The prevalence of fungi in collected biological materials from chronic dialysed patients, treated conservatively and persons from control group was determined at the level of 96.4%, 64.0% and 63.3%, respectively. In all examined groups fungi were found in high percentage in the ontocenoses of oral cavity and gastrointestinal tract; in dialysed patients (40.7%), treated conservatively (34.4%) and in persons with negative history of any renal disease (42.1%). It should be expressed that in 9.37% of patients with chronic renal failure undergoing conservative treatment fungi were determined at the same time in materials obtained from three ontocenoses--oral cavity, gastrointestinal tract and urinary tract. The majority of C. albicans strains (34.0%) was described by the use of code AUX 2 576 174; code 2 566 174 was found in 23.4% of species. Strains with code AUX 2 576 174 in 96,9% were isolated from chronic haemodialysed patients whereas code 2 566 174 characterized strains (68.2%) obtained from patients treated conservatively. Examined fungal strains from genus Candida obtained from all persons revealed the activity of 10 to 17 enzymes in the API ZYM test. Strains of genus Candida albicans obtained from patients with chronic renal failure treated conservatively and undergoing haemodialysis in majority belonged to biotype A (58.8%) and D3 (40.4%), respectively. PMID- 17912813 TI - [The usefulness of ELISA test for early serological detection of Trichinella spp. infection in pigs]. AB - Trichinellosis is a parasitic zoonosis transmitted to humans through consumption of raw or undercooked meat from animals infected with nematodes of the Trichinella genus. Every year seropositive cases are found among the human population and thus trichinellosis still remains an epidemiologically important disease. The first step of the study was the optimization of a new ELISA method enabling an early and specific serological diagnosis of trichinellosis in pigs and wild boars using excretory-secretory (ES) antigens obtained from in vitro cultures of L1 T. spiralis. Serum samples were assayed for anti-T. spiralis IgG antibodies using the new ELISA protocol and a reference test--Standard manufactured by Institut Pourquier. The optimization involved the selection of suitable plates for antigen coating, dilution of sera and antibodies and their time of incubation. On the basis of the optimization a new ELISA procedure for the detection of IgG and IgM against T. spiralis was elaborated. Conventional, Iberian pigs and SPF (Specific Pathogen Free) pigs were infected with 200, 1000 and 20,000 muscle larvae of T. spiralis. Serum samples were obtained at 5 and 1 dbi (day before infection), and 5, 10, 15, 20, 25, 30, 40, 50, 60 dpi (day post infection) and screened for specific IgG antibodies against excretory-secretory L1 T. spiralis antigens. Serum samples were obtained from the EU project Trichiporse: "Safe pork and horse meat on EU markets: early and unbiased diagnostic tests for Trichinella". Field samples of conventional pigs (1474) and wild boars (1784) were obtained from slaughter houses in different parts of Poland. Pigs were examined for the presence of Trichinella spp. using the artificial digestion method. Only four pigs were naturally infected with T. spiralis, the remaining were Trichinella larvae free. ELISA was used to examine IgG levels against L1 T. spiralis in pig and wild boar sera. The usefulness of ELISA for anti-IgG detection in pigs is usually limited by the nature of the antigen. The antigens were prepared in different laboratories: in Germany--Ag ES L1 T. spiralis (N), Italy--Ag ES L1 T. spiralis (W)) and in Poland--Ag ES L1 T. spiralis. Cut-off values for ELISA along with the estimated sensitivity and specificity were calculated using different methods: S/P%, M+3SD and ROC (Receiver Operating Characteristic). In SPF and Iberian pigs inoculated with 200, 1000 and 20,000 L1 T. spiralis, specific antibodies were detected 40, 30 and 25 dpi, respectively, with the use of the Standard (reference test). The analysis of the two ELISA procedures demonstrated a high sensitivity and specificity for the newly elaborated test utilizing the Ag ES L1 T. spiralis. In conventional pigs infected with 20,000 L1 T. spiralis specific antibodies were detected from 20 dpi when employing the new protocol. Similar results for the Standard and new ELISA test were obtained for serum samples of conventional pigs infected with 200 and 1000 larvae, which became positive from 40 dpi and 30 dpi, respectively. The results showed that both: the Standard and new protocols were comparable, and based on this, the new test was applied for further research. Results obtained adopting the new protocol with three antigens showed that two of them: Ag ES L1 T. spiralis (W) and Ag ES L1 T. spiralis are similar. The specific IgG antibodies for infective doses of 200 and 1000 larvae for these antigens were detectable 40 and 30 dpi respectively. In pigs infected with the highest dose of T. spiralis larvae IgG antibodies were detectable from 20 dpi when Ag ES L1 T. spiralis was used. These results strongly indicate that in examined pigs, the specific IgG response to T. spiralis infection is dose dependant. Of 1474 examined pig sera only 0.99% gave a positive signal against ES L1 T. spiralis antigen. Of 1784 examined wild boars sera only 0.68 % gave positive results using the new ELISA protocol. ELISA is a useful method for detecting specific IgG antibodies in pigs experimentally infected with different doses of T. spiralis and naturally infected pigs. In pigs the specific IgG response is dose dependant. The Ag ES L1 T. spiralis increases the specifity of the method and reduces false positive results. Simultaneous use of both methods: digestion and ELISA for the diagnosis of Trichinella in naturally infected pigs and wild boars may increase the chances of eliminating meat infected with T. spiralis larvae. PMID- 17912814 TI - [Embryonic development of the cestode Mosgovoyia ctenoides (Anoplocephalidae)]. AB - In this study the cleavage divisions and the ultrastructural analysis of early embryos as well as cellular organisation of infective oncosphere of the anoplocephalid cestode Mosgovoyia ctenoides are described. The early cleavage is unequal and results in the formation of three types of blastomeres: 2 large macromeres containing large electron dense granules, 3 medium-size mesomeres and several small micromeres. In the early stage of oncospheral morphogenesis, formation of three following primary embryonic envelopes takes place: (1) the capsule replaced by thick, rigid outer coat originated form the uterine material secretion, (2) the outer envelope and (3) the inner envelope. The capsule is formed from the vitellocyte material. Two macromeres contribute to the formation of the outer envelope and three mesomeres take part in the formation of the inner envelope. The inner envelope undergoes differentiation into three sublayers: (1) a thick extraembryophoral cytoplasmic layer, (2) an electron-dense embryophore, as a stiff pyriform apparatus, and (3) a thin intraembryophoral cytoplasmic layer containing mesomere nuclei. The oncosphere is located in the extended cupule-like part of the pyriform apparatus. Four egg envelopes surround the mature infective oncosphere of M. ctenoides: (1) a thick outer coat, (2) the outer envelope, (3) the inner envelope with a characteristic pyriform apparatus and (4) the oncospheral membrane. Hook morphogenesis takes place inside six symmetrically arranged oncoblasts, each of which shows a characteristic large nucleus of semi lunar shape. At the beginning the "hook-forming center" appears in the cytoplasmic part of each oncoblast. It consists of numerous free ribosomes, polyribosomes, mitochondria and Golgi complexes. The hook-forming center is involved in synthesis of a hook primordium, which undergoes differentiation and elongation into the fully developed hook. Mature hook consists of three parts: (1) blade, (2) shank, (3) base, and at the site of its protrusion from the oncosphere, is surrounded by a circular septate junction. Wide bands of hook muscles are attached to the basal and collar parts of the hook. The hook blades project outside the oncospheral body into a large cavity that is delimited by the hook region membrane. In the fully developed oncosphere of M. ctenoides three pairs of oncospheral hooks together with specialized hook muscles form a complex of "hook muscle system", responsible for coordinated hook action. The surface of the infective oncosphere is covered by a thin cytoplasmic layer of oncospheral tegument connected with the so-called "binucleate subtegumental cell", situated deeper in the oncospheral body. Below the cytoplasmic layer are situated wide bands of the somatic musculature responsible for oncospheral body movements. Five major types of oncospheral cells have been distinguished in the infective oncosphere: (1) a binucleate subtegumental cell, (2) a binucleate penetration gland, (3) two nerve cells, (4) numerous somatic cells, and (5) six germinative cells. During development of the oncosphere, changes in the concentration of glycogen and number of lipid droplets were observed. In the early embryos glycogen particles were most abundant in the macromere cytoplasm, whereas in micromeres concentration of glycogen was observed to be lower. In the course of the differentiation of the oncospheral envelopes glycogen was progressively distributed to other parts of the developing embryo. Simultaneously, a great increase in the number of lipid droplets was detected. However, during the preoncospheral phase of development a progressive reduction of lipid droplets was observed. This may indicate that lipids play a role of the energy source for developing oncosphere. PMID- 17912815 TI - [Regulation of the immune response in BALb/c mice infected with Heligmosomoides polygyrus]. AB - The aim of the studies was to identify the regulatory mechanisms that act at different levels of the ongoing immune response in BALB/c mice infected with intestinal nematode H. polygyrus. The role of TGF-beta during the course of H. polygyrus infection and an immunosuppressive action of the nematode against eosinophil response in allergic pulmonary inflammation has been studied. An attempt to identify the immunoregulatory proteins of the parasite has been performed as well. The obtained results proved: (1) for the first time the direct role of TGF-beta in the regulation of the immune response during helminth infections. Neutralization of TGF-beta in vivo increased concentration of IL-12, TNF-alpha and IL-10 in serum of infected mice and restored the control number of eosinophils in the intestinal mucosa. The mobilization of the immune response after neutralization of TGF-beta led to persistent decrease of nematode egg production and faster rejection of the worm from mouse intestine; (2) for the first time it was shown that the reduction of eosinophil number was due to the lower production of eotaxin and reduced expression of CCR3 receptor, playing an essential role in the chemotaxis of these leukocytes in Ova-related asthma; (3) significant decrease of T cell proliferation by one of the H. polygyrus protein fraction. With the use of mass spectrometry seven proteins have been identified: two heat shock proteins, disulfide isomerase, calreticulin, calumenin, fructose bisphosphate aldolase, glyceraldehyde-3-phosphate dehydrogenase. From the bibliographic data it may be supposed that calreticulin could mediate the downregulation of lymphocytes proliferation. The fraction with calreticulin stimulated also production of specific IgE. PMID- 17912816 TI - Enjoying the commute. PMID- 17912817 TI - ICD-9 changes: new codes of note. PMID- 17912818 TI - The ideal medical practice model: improving efficiency, quality and the doctor patient relationship. PMID- 17912819 TI - 7 strategies for creating a more efficient practice. PMID- 17912820 TI - Quick ways to maximize your office space. PMID- 17912821 TI - The medical home: an idea whose time has come ... again. PMID- 17912822 TI - Rewarding failure. PMID- 17912823 TI - [Adaptive properties of Trichnella spiralis isolate from Northern Ossetia]. PMID- 17912824 TI - [Experimental model of activated Lamblia (Giardia) muris infection in albino mice]. AB - Experimental L. muris infection was reproduced in 100% of the intact albino mice intragastrically given levomycin in an average total dose of 15.88-34.84 or 0.88 1.02 g/kg for 18-34 days. With levomycin administration, the intensity of giardiasis was 1121.6-8540.1 (mean 4830.9) thousand L. muris trophozoites per animal. The total number of trophozoites per animal decreased to 302.2-3481.4 (mean 1546.4) thousand and 28.1-324.0 (mean 109.4) thousand specimens 5-8 and 11 13 days after discontinuation of the antibiotic, respectively. The maximum number of L. muris trophozoites was observed in the proximal and middle portions of the murine small intestine during and after the administration oflevomycin. The highest isolation of cysts was seen 12-14 days after the initiation of administration of the antibiotic. Following 8-10 days of terminations of a course of levomycin therapy the native smear of animal feces showed no Lamblia cysts. In mice with activated infection, the isolation rate of Lamblia cysts was directly related to the intensity of intestinal infection with trophozoites of the parasite. PMID- 17912825 TI - [Typification of opisthorchiasis foci in the Orenburg Region]. AB - On making expedition and studying the functioning of opisthorchiasis foci in the Orenburg Region, the authors have districted the region and identified 3 types of districts according to the pattern of Opisthorchis transmission: 1) areas lacking the conditions for forming an invasion foci; 2) those having all links of the focus, but lacking pathogen circulation; and 3) those having all conditions for pathogen circulation and the human beings are infected from the local infested fish--Type C. By studying opisthorchiasis in the Orenburg Region, the authors have established the rate of morbidity growth and their nonuniform distribution in this region. The findings have formed the basis of dividing the Orenburg Region by the endemicity of opisthorchiasis into 4 types of areas: non-endemic, low, moderate, and high endemic. The results of districting the region by the endemicity of areas, the nature of foci, and the social and natural factors determining an epidemiological process have formed the basis of a differential package of preventive measures against opisthorchiasis in the Orenburg Region. PMID- 17912826 TI - [Rivers of the Orenburg Region: maintenance of the life cycle of Opisthorchis felineus]. AB - Specific hydrological and climatic features contribute to the functioning of the parasitic system egg--mollusk--fish--man parasitic in the Orenburg Region. Four types of areas, such as floodplain, suprafloodplain-terrace, valley-ravine, and near-lake ones, are adequate for the formation of a natural opisthorchiasis focus. The periods of active functioning of a natural opisthorchiasis focus have been established in the Orenburg Region. These include the emergence of the first cercariae and, therefore, the first (in this epidemic season) infection of the supplementary host (fish) occur in mid-June to early July and then 6 weeks later metacercariae achieve invasiveness in the fish muscles and subcutaneous fat, as a result the first infection of the final host with Opisthorchis occurs in early August. PMID- 17912828 TI - [Parasitocenosis as a biotic factor influencing oncogenesis and the epizootic situation]. PMID- 17912827 TI - [Effects of some species of hydrocoles on the development of ascarid eggs]. AB - The study established that aquatic plants differently affected the development of ascarid eggs. Some species of algae exerted a damaging effect on ascarid eggs only at early stages of embryogenesis while others did at its late stages, but some species did not produce any noticeable effect on the development of the eggs. A great difference was found in the development of ascarid eggs in different seasons. Gas exchange conditions were ascertained to affect the development of ascarid eggs. Analysis of the effect of egg lysozyme versus mollusk one on ascarid embryogenesis demonstrated that the latter had a more pronounced antiparasitic activity. PMID- 17912829 TI - [The infection rates of echinococcosis and cysticercosis in agricultural animals in the Orenburg Region]. AB - The infection rates of echinococcosis and cysticercosis in cattle and pigs were studied in the Orenburg Region. Those of echinococcosis remained high (the extensiveness of invasion increased from 6.7 to 23.8%, as shown by the 1996-2001 data) despite a considerable reduction in the animal numbers in the 1990s. The infection rate of cysticercosis in the cattle with cysticercosis was much lower (the extensiveness of invasion increased from 0.12 to 0.27% over this period). The lowest infection rate was established for porcine cysticercosis (the extensiveness of invasion was 0.1%). PMID- 17912830 TI - [Parasitic diseases in fishes habiting the water reservoirs in the Orenburg Region]. AB - The material to be analyzed has been provided by the Orenburg regional veterinary laboratory as work reports. The study material units (fish, autoptic data) totaled 11452 specimens over 2001-2005. Two major groups of diseases--protozoan diseases and helminthism--have been identified. Analysis of the studies suggests that the parasitological state of the water reservoirs is complicated in the Orenburg Region. PMID- 17912831 TI - [Role of various Ixodes ticks as vectors of the causative agents of tick-borne encephalitis and borrelioses in the Orenburg Region]. PMID- 17912832 TI - [Hematozoon cenoses of small rodents and insectivora in the Orenburg Region]. AB - A total of 2942 specimens of 15 species of ground rodents and insectova in the Orenburg Region were caught and examined during long-term studies. The investigators detected 7 taxonomic groups of hematozoons: rickettsia (Anaplasma sp., Grahamella sp., Haemobartonella sp.), protozoa (Trypanosoma sp., Plasmodium sp., Piroplasma sp.), and nematodes (Filariidae spp., larval stages). The authors give information on the species composition and infection extensiveness of individual systematic groups of small mammals, the most important morphometric and biological signs of blood parasites, and the specificity of parasite-host relations. The Eversmann hamster was found to have parasitic protozoa of the genera Trypanosoma and Piroplasma, which had not been earlier described in the scientific literature. PMID- 17912833 TI - [Fleas of small mammals in the steppe zone of the Southern Urals]. PMID- 17912834 TI - [The rate of infection with hematozoons in birds in relation to season]. PMID- 17912835 TI - [Animal helminthisms and their zonal prevalence in the Orenburg Region]. AB - In the 1980s, porcine ascariasis, animal strongyloidiasis, and ruminant moniesiases were commonest in the Orenburg Region. At present, among the nosological entities there is a preponderance of the same helminthisms, i.e. they have the pattern of steady-state invasions. The strongyloidiases are spread in all zones of the region, moniesiases of large and small cattle are in the central and eastern zones, respectively. PMID- 17912836 TI - [Short-horned flies (Diptera: Brachycera) are ectoparasites of warm-blooded animals]. AB - In the Orenburg Region, the fauna of blood-sucking short-horned flies comprises 23 species, including 12 obligate hematophagi and 11 facultative ones. The collected flies belong to the families Tabanidae and Muscidae. Dominant species, their spread, environmental characteristics, and phenological features have been established. The ratio of various types of blood-sucking flies has been analyzed. PMID- 17912837 TI - [The fauna of Ixodes ticks in the Saratov Region under the present conditions]. AB - A total of 10566 specimens of 10 species of Ixodes ticks, such as Dermacentor marginats, D. reticulatus, D. niveus, Rhipicephalus rossicus, Ixodes ricinus, I. persulcatus, I. crenulatus, Haemaphysalis punctata, Hyalomma scupense, and H. marginatum, were collected and identified in the Saratov Region in 2002-2006. Dermacentor marginats and D. reticulatus were found to be the most dominant species. Rhipicephalus rossicus, Ixodes ricinus, and Hyalomma scupense were also constantly detected in the region. I. persulcatus considered earlier untypical of the Saratov Region was detected. The paper presents data on the confined geographical and environmental features of individual species of Ixodes ticks in the Saratov Region. PMID- 17912838 TI - [Localization of larvae of Neotrombicula (N) monticola (Trombiculidae) ticks on the vertebrates]. AB - The specific features of distribution of Neotrombicula (N) monticola Schluger et Davidov, 1967 on small mammals were studied in the Tien Shan montains (Kirghiz ridge). N. (N ) monticola was found to occur in all the places under study. Nine species of mammals (pigmy white-toothed shrew, dwarf hamster, tamarisk gerbil, Turkestan rat, long-tailed mouse, Tien Shan, Kirghiz, and silver voles, and wood mouse) were established to be feeders of larvae of the ticks. N. (N) monticola larvae were detected in three topographic zones and seven portions of the body of vertebral hosts. The inner auricle is the major site of tick attachment to the host. Preference of N. (N) monticola in selecting the host is likely to be based on the morphophysiological features of partners. PMID- 17912839 TI - [Evaluation of the antihymenolepidous activity of cyclarin]. PMID- 17912840 TI - [Antibodies in trichinosis and duration of their detection in convalescents]. PMID- 17912841 TI - [Detection of coproantigens in experimental trichinosis]. PMID- 17912842 TI - [A case of human dirofilariasis in the Astrakhan Region]. PMID- 17912843 TI - [Current antimalarial drugs recommended by the World Health Organization for the treatment and prevention of malaria]. PMID- 17912844 TI - [Classic of physiology (on the 125th birthday anniversary of academician Leon Abgarovich Orbeli)]. PMID- 17912845 TI - [The contribution of academician Leon Abgarovich Orbeli and his school to the development of physiology of extreme conditions]. AB - The paper is devoted to the role of Academician L. A. Orbeli and his school in foundation and development of extreme physiology. The works of L. A. Orbeli, his pupils and collaborators contributed to the advancement of key theoretical and applied aspects of hyperbaric physiology, flights in stratosphere, aviation physiology and medicine, space physiology and radiobiology. PMID- 17912846 TI - [School of the school of Leon Abgarovich Orbeli]. AB - In 2007, L. A. Orbeli would have been 125. He was distinguished by extremely wide scientific interrests; he created one of the most numerous and fruitful scientific schools. He authored prominent achievements in physiology of autonomic nervous system, evolutionary physiology, sensory physiology, renal physiology, physiology of underwater labor. Orbeli paid much attention to the scientific organizational activity, he was academician-secretary of the Division of Biological Sciences of the USSR Academy of Sciences, President of the Society of Physiologists, Biochemists and Pharmacologists, Editor-in-Chief of the USSR Physiological Journal, etc. Principles of the scientific scholl founded by Orbeli are service to science and society, propity to scientific ethics, humanity. PMID- 17912847 TI - [The interaction between glycine and GABA postsynaptic effects in the spinal cord neurons of frog Rana temporaria]. AB - Patch-clamp study in the whole multipolar cell (presumably motoneuron) was performed, the cells having been mechanically isolated from the spinal cord of the frog Rana ridibunda. It was shown that GABA and glycine, when applied simultaneously, produced a transmembrane current. Its amplitude was lower than the summed amplitude of currents produced in the same neuron by separate applications of GABA and glycine. The investigation of this occlusion showed that the superfusion of the neurons with solution containing 0.2 mM of glycine totally blocked the responses to GABA (5 mM) application, and vice versa. The crossinhibition can lie in the basis of this phenomenon. It could be due to either the existence of a common receptor complex sensitive to both GABA and glycine or to interaction between GABA and glycine receptors. PMID- 17912848 TI - [Aldosterone: the mechanisms of molecular action]. AB - Aldosterone: a steroid hormone of adrenal cortex, has recently attracted much interest not only due to its great importance in regulation of salt and water balance, but also because of its key role in therapy of cardiovascular and renal pathology. The classical genomic mechanism of molecular action of aldosterone is mediated through interaction with mineral-corticoid receptors. Fast nongenomic pathway of cell signal transduction begins with interaction with hypothetic membrane receptors and includes activation of different kinase cascades. Interference of these two pathways of signal transduction assures abroad spectrum of aldosterone effects depending on the cell type, and also secures multycomponent regulation depending on the need of specific functional and stress situation. This review is dedicated to modern views of mechanisms of aldosterone molecular action, mostly of the level of aldosterone-sensitive segment of kidney nephron. PMID- 17912849 TI - [Prenatal ontogenesis of the human brain cortex temporal area]. AB - Prenatal ontogenesis of temporal areas of the human cortex was studied. In the fetal cortex at the gestational age of 16-18 weeks three zones can be distinguished: marginal zone (eI layer), cortex plate and subplate. At 20-26 weeks cortex plate is divided into following layers: eII, eIII, eIV, eV and eVI, with "efferent" complex of layers being wider than "associative" one. The subplate neurons are eliminated in the fetus at 27-33 weeks, when "associative" complex composes over 50 per cent of the cortex thickness. The subplate has been identified by positive correlation between layer eII and the upper subplate layer spu cell density. PMID- 17912850 TI - [Involvement of striatal serotonin in fluoxetine effects on adrenocortical function and behaviour]. AB - Treatment of the adult rats with selective serotonin (5-HT) reuptake inhibitor: fluoxetine and its complexes with glycyrrizhinic acid during 2 weeks (25 mg/kg/day) significantly increased plasma corticosterone levels that were measured after 5-min plus-maze. All the drugs decreased the content of 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) in the striatum as well as 5 HT in the hippocampus. There was a significant negative correlation between 5-HT in the striatum and corticosterone levels. These data suggest that fluoxetine induces serotoninergic changes in the striatum that might be related to neuroendocrine and behavioural effects of the drug. PMID- 17912851 TI - [Mismatch negativity in the auditory event-related potentials in response to abrupt and smooth displacements of virtual sound source]. AB - The work investigated event-related potentials, mismatch negativity (MMN), and P3a component under dichotic stimulation with deviant stimuli simulating abrupt or smooth displacement of auditory images to the left or to the right from the head midline by means of interaural time delay introduced into the deviant stimuli. Repetitive standard stimuli were localized near the head midline. All deviant stimuli elicited mismatch negativity and P3a component. It was shown the MMN for smooth deviant motion was lower than that for the abrupt deviant displacement. MMN amplitude for both deviant types obviously depended on interaural time delay, which confirms that MMN might be considered as a measure of the auditory system spatial discriminative ability. The P3a component demonstrated the same amplitude dependences as the MMN. The results obtained are discussed in respect to manifestation of the processes underlying the auditory motion detection in the event-related potentials. PMID- 17912852 TI - [Age-related characteristics of relationships between brain blood flow, liquor dynamics and biomechanical properties of human cranium]. AB - The peculiarities of relationships between changes of cerebral blood flow, intracranial liquor dynamics and skull biomechanics in humans were studied in an age aspect. For this aim, a non-invasive method was proposed based on concomitant registration of rheoencephalogram and transcranial dopplerogram and evaluation of relationships between intracranial volume and pulse pressure changes (P-V index). The data obtained were analyzed by pattern-phase computer processing and compared with the blood flow parameters. The investigation was carried out on healthy volunteers of 18-25, 40-50 and 65-75 years of age. It was shown that circulatory metabolic supplying of human brain was supported by such factors as volume brain blood flow, intracranial liquor dynamics in cooperation with skull biomechanics. The cerebral blood flow decrease at aging could be compensated by increase of the reserve-compensatory abilities of the system of cranial-spinal liquor dynamics. PMID- 17912853 TI - [Analysis of electrical activities of the junction region of the ureter with bladder and surrounding zones in rats]. AB - Spontaneous electrical activity of the rat ureters per bladder zone was studied. In area of ureter connection with bladder, slow-wave activity was revealed. In bladder bordering ureter zone, rapid spike activity was found that was fully correlated with the genesis of upper located fluctuation processes. PMID- 17912854 TI - [History of physiology in 18th and early 19th centuries]. PMID- 17912855 TI - Orchard fresh. Greenfield site provides home for newest Kaiser facility. PMID- 17912856 TI - Form follows function. Improving designs by understanding clinical operations. PMID- 17912857 TI - Rain out. Getting operating room humidity under control. PMID- 17912858 TI - Factors of sustainability. Gauging environmental impact when deciding whether to build or renovate. PMID- 17912859 TI - [The patient definitely wants a urinary catheter. Where from comes the marble pattern on the abdomen?]. PMID- 17912860 TI - [Conflicts in the treatment of terminal patients. Between physician's duty to help, patient's will and assisted death]. PMID- 17912861 TI - [Recurrent vomiting. In this case it's not stomach's fault]. PMID- 17912862 TI - [Benign prostatic hyperplasia: background and diagnosis]. AB - Lower UrinaryTracts Symptoms (LUTS) due to Benign Prostatic Hyperplasia (BPH) represent an increasing prevalent condition in ageing men. Patients often seek primarily consultation at their general practitioner. Aetiology and natural history of LUTS due to BPH have not been completely clarified. The development of symptomatic LUTS is age-dependent and determined to varying degrees by the presence of Benign Prostatic Hyperplasia (BPH), Benign Prostatic Enlargement (BPE) as well as Bladder Outlet Obstruction (BOO). A causal relationship does not always exist. Basis for a specific medical or surgical treatment in the individual patient with LUTS due to BPH is an exact diagnosis by the practising urologist. PMID- 17912863 TI - [Benign prostatic hyperplasia: medical therapy]. AB - Primary aims of the medical therapy for BPH are improvement of subjective symptoms and quality of life as well as the prevention of long-term complications such as acute urinary retention and renal failure. Secondary goal is inhibition of disease progression. The medical therapy should be tailored to each patient according to the individual complaints and risk of progression. Plant extracts, alpha-blockers and 5-alpha reductase inhibitors represent the most common prescribed substances. Recent data suggest beneficial effects for the use of antimuscarinic agents and phosphodiesterase type 5 inhibitors. PMID- 17912865 TI - [Fibrosarcoma of the female breast]. PMID- 17912864 TI - [Benign prostatic hyperplasia: surgical treatment]. AB - According to the current treatment algorithm, surgical treatment of benign prostatic syndrome (BPS) is the next step after a pharmacological treatment. In addition to conventional transurethral resection of the prostate (TURP), much interest is shifting towards alternative sugical techniques such as transurethral microwave therapy (TUMT),transurethral needle ablation of the prostate (TUNA) and several laser techniques. PMID- 17912866 TI - [New European guidelines--risk analysis inherent in diagnosis of hypertension]. PMID- 17912867 TI - [You should think it over on time. How much is your practice still worth today?]. PMID- 17912868 TI - [Decision of the Joint Federal Committee. Cancer prevention and health examinations remain voluntary]. PMID- 17912869 TI - [History of mumps]. PMID- 17912870 TI - [Disease from Work?]. PMID- 17912871 TI - [Renin inhibitors. Aliskiren for the treatment of high blood pressure]. PMID- 17912872 TI - [Thujone]. AB - Thujone is an ingredient of essential oils of different herbs and causes exciting effects in the central nervous system. Absinthe lkiquor with thujone was forbidden in most European countries because it was taken in abuse. Up to now there have been a lot of theories about the mechanism of the pharmacological effect on the central nervous system. New researches verify an interaction of thujone with GABA-receptors. Within the process of adapting the EU-law the absinthe liquor is allows again and is not classified as dangerous when taken as directed, as the content of thujone should not exceed the particular limits depending on content of alcohol in the liquor. PMID- 17912874 TI - [Head lice (pediculosis capitis). Advice on infectious diseases--a leaflet for the physician]. PMID- 17912873 TI - [Sialorrhea--causes and treatment options]. AB - Sialorrhea (drooling) is the involuntary spillage of saliva from the mouth. Drooling of saliva appears to be the consequence of a dysfunction in the coordination of the swallowing mechanism, resulting in excess pooling of saliva in the anterior portion of the oral cavity and the unintentional loss of saliva from the mouth. There are specific and symptomatic approaches to manage this condition. Treatment options range from conservative (i.e., observation, postural changes, biofeedback, motoric therapy of the mouth, acupuncture) to more aggressive measures such as medication, radiation, and surgical therapy. PMID- 17912875 TI - [Splitting of tablets: small pieces a risk]. AB - For economic reasons physicians prescribe more and more multiunit tablets. Splitting of multiunit tablets depends on the physical-chemical properties of the agents, the galenic of the dosage form, the size and contour of the tablet and the shape of the score. Tablets with one or more scores are prepared to be divided for a single/multiple dose. How easily and exact a tablet can be divided depends heavily on the physical shape, its size and the outfit of the score. The fragments have to fulfil the requirements according to the European Pharmacopoeia: Uniformity of multiunit tablets. Since exact dosing is guaranteed only if tablets are divided properly, information and guidance of the patients by the physician and pharmacist is of critical importance. PMID- 17912876 TI - [When does angina come from scarlet fever?]. PMID- 17912877 TI - [Analgesics. Self medication for headaches]. PMID- 17912878 TI - [Aspirin. Treatment of migraine and tension headaches]. PMID- 17912879 TI - [Acute migraine. Sumatriptan-naproxen combination tablets work better than a single substance]. PMID- 17912880 TI - [Juvenile rheumatoid arthritis. What is the optimal treatment?]. PMID- 17912881 TI - [Shark cartilage extract. No effect on bronchial carcinoma]. PMID- 17912882 TI - [Alcohol consumption. Moderate doses reduce total mortality]. PMID- 17912883 TI - [Warts. Occlusion therapy with duct tape brings no advantage]. PMID- 17912884 TI - [The metabolic syndrome]. AB - The "metabolic syndrome" consists of some common risk factors for cardiovascular diseases: central obesity, diabetes mellitus, hyperlipidaemia and hypertension. The metabolic syndrome (MTS) leads to increased morbidity and mortality and to higher direct and indirect healthcare costs. The MTS can be diagnosed using the NCEP/ATP III criteria. The prevalence of the MTS in Germany is estimated at 23.8% and is expected to rise further, due to increasing obesity among children and adolescents. Studies have shown that the MTS leads to increased cardiovascular and total mortality and thus to a decreased life expectancy. Studies estimating the total costs of MTS are missing, but after addition of the costs for all the risk factors it is assumed that MTS costs amount to 5% of total healthcare costs. This is the result of the more frequent demand of health services and longer hospitalisation of patients with MTS. Even moderate weight loss can decrease the rates of morbidity and healthcare costs. PMID- 17912885 TI - [Reasons for different loss experiences with men and women in private disability insurance]. AB - Women in general have a more unfavourable loss experience in private disability insurance due to biological and social reasons (enhanced rate of malignant and psychic diseases). Contrary to the enhanced rates of orthopaedic diseases and accidents with men, women's diseases are not work-related and therefore are not to be compensated by any extra amounts concerning occupational groups. In spite of the enhanced risk of occupational disability, women are more seldom insured against this risk than men. So--despite the rising female rate--there is still a clear backlog demand. PMID- 17912886 TI - [Integrated case fees in cardiosurgery--a pilot project for fast-track rehabilitation]. AB - BACKGROUND: Lengthy recovery and treatment times following cardiosurgical interventions were the motivation for introducing a pilot procedure to integrate acute and rehabilitative treatment structures. The advantage of such a pilot procedure is the medico-economic link between direct transition from acute care to rehabilitation treatment and cutting average case costs. With this in mind, shared case fees for patients following cardiosurgery are being agreed in a pilot project between health insurance companies, acute-care hospitals and rehabilitation clinics. The aim of this study was thus to investigate whether rehabilitation directly after cardiosurgery without prior transferral to an acute care hospital is comparable with the conventional procedure involving acute care. METHODS: A total of 221 patients were included in the investigation. The pilot project group comprised 159 patients (mean age 70 +/- 6 yrs, 117 men and 42 women) who were transferred directly to rehabilitation following cardiosurgery. The control group, comprising 62 patients (mean age = 71 +/- 6 yrs, 42 men and 20 women), was transferred to an acute-care hospital following cardiosurgery before commencing rehabilitation. Sociodemographic and clinical data were comparable between the two groups. RESULTS: At the end of rehabilitation, the mean maximum ergometric performance in the pilot group was 96 +/- 33 W, significantly higher than the control group's performance of 81 +/- 31 W. One difference between the two groups related to complications. During rehabilitation, complications occurred more frequently within the pilot group. In the pilot group, compared to the control group, postcardiotomy syndrome occurred in 45.3 versus 25.8% and impaired wound healing in 10.1 versus 4.8% of cases. Despite these results, the pilot group demonstrated a significantly shorter overall hospital stay of 39.5 +/ 7.5 days compared to the control group stay of 45.7 +/- 9.7 days. CONCLUSION: Compared to the control group, the pilot group was at no disadvantage with regard to clinical or performance data by the end of rehabilitation. Cardiac complications occur more often during rehabilitation taking place directly after cardiosurgery than with the conventional procedure. These can be viewed, however, as complications occurring directly in temporal conjunction with the operation and as to be expected. Complications attributed directly to fast-track rehabilitation can be excluded. In the pilot group the overall hospital stay was thus shortened. In an environment of legislative restructuring within the healthcare sector, this shows that adequate treatment of cardiosurgical patients is still guaranteed with fast-track rehabilitation. PMID- 17912887 TI - [Predictive genetic testing for hereditary breast cancer and hereditary nonpolyposis colorectal cancer]. AB - Genetic tests are no longer used exclusively for diagnostic purposes, but they serve in increasing numbers to assess the predisposition for illnesses. Predictive genetic testing of this kind includes screening for genetic predispositions towards tumours. Of special significance are familial breast and ovarian cancer caused by BRCA1 - and BRCA2- mutations and hereditary nonpolyposis colorectal cancer (HNPCC). The observation of such conditions constitutes a considerable burden for the individuals concerned as well as their families. So far it has not been sufficiently established to what extent methods of prevention and early diagnosis can offer effective protection against the risk of tumours. While prophylactic organ resections lower the morbidity and mortality associated with cancer, they are also responsible for a loss of physical and emotional well being. Therefore, an awareness of hereditary predispositions towards tumours causes considerable need for competent medical advice and clinical research. Interdisciplinary projects can offer the suitable organisational base for these tasks. Doctors, universities and health insurances are faced with the challenge of developing and financing corresponding structures of medical care and research. PMID- 17912888 TI - [Quo vadis DMP? The chronic-program in the private health insurance on the way to "Chronic Care Management"]. PMID- 17912889 TI - [Targeting medical insurance educational offerings]. PMID- 17912890 TI - [PKV-Ombudsman: resolving conflicts creatively ]. PMID- 17912891 TI - [Development of vaccines against the next pandemic influenza virus]. PMID- 17912892 TI - [Origin and evolution of the terms "german" and "German" (II)]. PMID- 17912893 TI - [Clarification of intermittent diarrhea using Positron-Emission-Tomography (PET) and CT-X-ray]. PMID- 17912894 TI - A decade of patient satisfaction survey results. Lessons learned in a large multispecialty group practice. PMID- 17912895 TI - Can't get no (physician) satisfaction? PMID- 17912896 TI - Physician relations: now more than ever. PMID- 17912897 TI - Competition heats up for health care organizations seeking prestigious award. PMID- 17912898 TI - Creating a perfect hospital. PMID- 17912899 TI - Can you recognize the winds of change? AB - If you find yourself between a rock and a hard place during your career, there are some tough choices you will have to make. Read how one physician executive made these choices. PMID- 17912900 TI - The future of hospital medicine. PMID- 17912901 TI - A roadmap for quality improvement in physician offices. PMID- 17912902 TI - Is ethically justified nepotism in hiring and admissions in academic health centers an oxymoron? PMID- 17912903 TI - Insights for running smooth residency and fellowship training programs. PMID- 17912904 TI - A new model: outpatient pediatric diabetes improves care, profits and protocols for managing other chronic illnesses. PMID- 17912905 TI - 3 keys to improving hospital/physician relations. PMID- 17912906 TI - Hospital surveillance. PMID- 17912907 TI - I want to be an entrepreneur... but I don't know what business to start. PMID- 17912908 TI - Assault on the professional ethic: will professionalism survive? PMID- 17912909 TI - Malpractice stress and coping--the role of the physician executive. PMID- 17912910 TI - Do your due diligence as you evaluate potential employers. PMID- 17912911 TI - Physician executive finds Poland's health care system shares many of the same problems as the U.S. system. PMID- 17912912 TI - Is I.T. the key to preventing hospital infections? PMID- 17912913 TI - Giving transcription a trim. PMID- 17912914 TI - A new battery of tests for lab systems. PMID- 17912915 TI - Quadramed tries to turn corner. PMID- 17912916 TI - Connecting for clinical trials. PMID- 17912917 TI - EHR project is something special. PMID- 17912918 TI - Identifying at-risk patients. PMID- 17912919 TI - Is all this technology really making patients safer? PMID- 17912920 TI - The IMPROVE_A temperature protocol for thermal/optical carbon analysis: maintaining consistency with a long-term database. AB - Thermally derived carbon fractions including organic carbon (OC) and elemental carbon (EC) have been reported for the U.S. Interagency Monitoring of PROtected Visual Environments (IMPROVE) network since 1987 and have been found useful in source apportionment studies and to evaluate quartz-fiber filter adsorption of organic vapors. The IMPROVE_A temperature protocol defines temperature plateaus for thermally derived carbon fractions of 140 degrees C for OC1, 280 degrees C for OC2, 480 degrees C for OC3, and 580 degrees C for OC4 in a helium (He) carrier gas and 580 degrees C for EC1, 740 degrees C for EC2, and 840 degrees C for EC3 in a 98% He/2% oxygen (O2) carrier gas. These temperatures differ from those used previously because new hardware used for the IMPROVE thermal/optical reflectance (IMPROVE_TOR) protocol better represents the sample temperature than did the old hardware. A newly developed temperature calibration method demonstrates that these temperatures better represent sample temperatures in the older units used to quantify IMPROVE carbon fractions from 1987 through 2004. Only the thermal fractions are affected by changes in temperature. The OC and EC by TOR are insensitive to the change in temperature protocol, and therefore the long-term consistency of the IMPROVE database is conserved. A method to detect small quantities of O2 in the pure He carrier gas shows that O2 levels above 100 ppmv also affect the comparability of thermal carbon fractions but have little effect on the IMPROVE_TOR split between OC and EC. PMID- 17912921 TI - Fate of polychlorinated dibenzo-p-dioxins and dibenzofurans in a fly ash treatment plant. AB - To understand the fate of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in a fly ash treatment plant that used the Waelz rotary kiln process (hereafter the Waelz process), the samples of input and output media were collected and analyzed. The most important PCDD/F source in input mass was electric arc furnace (EAF) fly ash, which had a mean PCDD/F content of 18.51 ng/g and contributed more than 99% of PCDD/F input mass, whereas the PCDD/F input mass fractions contributed by the coke, sand, and ambient air were only 0.04%, 0.02%, and 0.000002%, respectively. For the PCDD/F output mass in the Waelz process, the major total PCDD/F contents of 43.73 and 10.78 ng/g were in bag-filter and cyclone ashes, which accounted for approximately 69% and 17%, respectively, whereas those of stack flue gas and slag were 14% and 0.423%, respectively. The Waelz process has a dechlorination mechanism for higher chlorinated congeners, but it is difficult to decompose the aromatic rings of PCDD/Fs. Therefore, this resulted in the accumulation of lower chlorinated congeners. The output/input ratio of total PCDD/F mass and total PCDD/F international toxicity equivalence (I TEQ) was 0.62 and 1.19, respectively. Thus, the Waelz process for the depletion effect of total PCDD/F mass was positive but minor, whereas the effect for total PCDD/F I-TEQ was adverse overall. PMID- 17912922 TI - Development of a local carbon dioxide emissions inventory based on energy demand and waste production. AB - This paper describes the study that led to the development of a carbon dioxide emissions matrix for the Oeiras municipality, one of the largest Portuguese municipalities, located in the metropolitan area of Lisbon. This matrix takes into account the greenhouse gas (GHG) emissions due to an increase of electricity demand in buildings as well as solid and liquid wastes treatment from the domestic and services sectors. Using emission factors that were calculated from the relationship between the electricity produced and amount of treated wastes, the GHC emissions in the Oeiras municipality were estimated for a time series of 6 yr (1998-2003). The obtained results showed that the electricity sector accounts for approximately 75% of the municipal emissions in 2003. This study was developed to obtain tools to base options and actions to be undertaken by local authorities such as energy planning and also public information. PMID- 17912923 TI - Protecting off-site populations and site workers from vapor discharges during shallow soil mixing at the North Carolina State University National Priorities List Site. AB - Although vapor monitoring is generally a component of remedial action activities, most sites do not have routine gaseous releases or vapor clouds erupting from the soil during implementation of the cleanup process (or during cleanup of the site). At the North Carolina State University Lot 86 National Priorities List Site, over 8410 m3 (11,000 yd3) of chemical waste was disposed at the Site, including organic solvents and shock-sensitive and air- and water-reactive compounds. During the Remedial Action, it was imperative to protect site workers and off-site populations from potential inhalation exposures. Engineering controls were incorporated into the shallow soil mixing process to limit the release of gaseous compounds. To quantify potential exposures to on-site and off site receptors, modeling was conducted to evaluate potential exposure routes and migration pathways. To demonstrate acceptable levels of airborne constituents, a multifaceted air sampling and monitoring program was implemented. To ensure that potential exposures could be quantified, passive dosimeters, continuous real-time monitoring, time-weighted whole air sampling, and grab samples of vapor clouds were all critical components of the air monitoring program. After the successful completion of the Remedial Action, the pre-Resource Conservation and Recovery Act (RCRA) chemical waste generated from the University's educational and research laboratories was entirely encapsulated and neither on-site workers nor off-site populations were exposed to analyzed compounds above any health-based action level (i.e., 15-min short-term exposure limit [STEL], 8-hr threshold limit value, or time-weighted average permissible exposure limit). PMID- 17912925 TI - Derivation of new emission factors for quantification of mass emissions when using optical gas imaging for detecting leaks. AB - This paper describes the development of new "leak/no-leak" emission factors that are suitable for estimating facilities' fugitive emissions when using an alternative work practice (AWP) that is based on optical gas imaging technology for detecting leaking piping system components. These emission factors were derived for valves, pumps, and connectors/flanges for instrument leak detection thresholds ranging from 3 to 60 g/hr using a combination of field data and Monte Carlo statistical simulation techniques. These newly derived leak/no-leak emission factors are designed to replace the U.S. Environment Protection Agency (EPA) 1995 Protocol factors, which were based on Method 21 monitoring of leaks at "uncontrolled" facilities. The emission factors published in the 1995 Protocol have not been updated since the 1970s. This derivation is based on results where the authors document the use of a Monte Carlo simulation technique to quantify the required leak detection thresholds that provide equal--or better- environmental benefits for an AWP. The use of these newly derived emission factors is demonstrated for different methods of computing fugitive emissions from a hypothetical model refinery. The resulting facility emissions calculated by using these new emission factors is compared with the existing emission estimation methods provided in the EPA 1995 Protocol. The results demonstrate that the new emission factors provide an emission estimate that is the closest to that obtained from the direct determination of total emissions by Monte Carlo simulations. PMID- 17912924 TI - Equivalent leak definitions for Smart LDAR (leak detection and repair) when using optical imaging technology. AB - Controlling fugitive emissions from leaks in petrochemical industry process equipment now requires periodic monitoring of valves, flanges, pumps etc., typically on a quarterly basis. Previous studies have shown that over 90% of the reducible emissions come from approximately 0.1% of the components, i.e. the large leakers. A new, and more cost-effective approach for controlling these large leakers would entail more frequent monitoring of process equipment, allowing for the detection and repair of the highly leaking components that contribute the most to emissions. This approach has been called "Smart LDAR." New optical imaging instruments, which significantly reduce monitoring costs, are now available to implement such an alternative work practice. This work describes the determination of the leak detection sensitivity (equivalency threshold) that an optical imaging instrument must achieve to ensure that it will provide at least the equivalent emission control of the current leak detection and repair practice. Equivalency thresholds were developed for various monitoring intervals. The U.S. Environment Protection Agency's Monte Carlo simulation approach was used to perform the analysis and to demonstrate that optical imaging, which is capable of identifying all of the largest leakers, can provide better control of fugitive emissions. PMID- 17912926 TI - Characterization of in-use light-duty gasoline vehicle emissions by remote sensing in Beijing: impact of recent control measures. AB - China's national government and Beijing city authorities have adopted additional control measures to reduce the negative impact of vehicle emissions on Beijing's air quality. An evaluation of the effectiveness of these measures may provide guidance for future vehicle emission control strategy development. In-use emissions from light-duty gasoline vehicles (LDGVs) were investigated at five sites in Beijing with remote sensing instrumentation. Distance-based mass emission factors were derived with fuel consumption modeled on real world data. The results show that the recently implemented aggressive control strategies are significantly reducing the emissions of on-road vehicles. Older vehicles are contributing substantially to the total fleet emissions. An earlier program to retrofit pre-Euro cars with three-way catalysts produced little emission reduction. The impact of model year and driving conditions on the average mass emission factors indicates that the durability of vehicles emission controls may be inadequate in Beijing. PMID- 17912927 TI - Establishment of a Box-Jenkins multivariate time-series model to simulate ground level peak daily one-hour ozone concentrations at Ta-Liao in Taiwan. AB - Box-Jenkins univariate autoregressive integrated moving average (ARIMA) and regression with time-series error (RTSE) models were established to simulate historical peak daily 1-hr ozone concentrations at Ta-Liao, Taiwan, 1997-2001. During 1995-2003, the 600 days of Pollution Standard Index (PSI) more than 100 (peak daily 1-hr ozone concentrations detected by greater than 120 ppm) at Tao Liao showed the highest ozone exceedances among the six monitoring stations in Kaohsiung County. To improve the predictability of extremely high ozone, two different principal components, PC1 and PC(1 + 2), were introduced in the RTSE model. Four typical predictors (particular matter with an aerodynamic diameter less than or equal to 10 microm, temperature, wind speed, and wind direction) plus a PC trigger remained significant in the RTSE model. The model performance statistics concluded that the RTSE model with PC1 was optimal, compared with the univariate ARIMA, the RTSE model without PC, and RTSE model with PC(1 + 2). The contingency table shows that the successful predictions of the univariate model were only 12.9% of that of the RTSE model with PC1. Also, the POD value was improved approximately 5-fold when the univariate model was replaced by the RTSE model, and almost 8-fold when it was replaced by the RTSE model with PC1. Moreover, introducing the PC trigger indeed enhanced the ozone predictability. After the PC trigger was introduced in the RTSE model, the POD was increased 69.9%, and the FAR was reduced 8.3%. The overall correlation between the observed and simulated ozone was improved 9.6%. Also, the first principal component was more useful than the first two components in playing the "trigger" role, though it counted only for 58.62% of the environmental variance during the high ozone days. PMID- 17912928 TI - Volatile organic compound emissions from dry mill fuel ethanol production. AB - Ethanol fuel production is growing rapidly in the rural Midwest, and this growth presents potential environmental impacts. In 2002, the U.S. Environmental Protection Agency (EPA) and the Minnesota Pollution Control Agency (MPCA) entered into enforcement actions with 12 fuel ethanol plants in Minnesota. The enforcement actions uncovered underreported emissions and resulted in consent decrees that required pollution control equipment be installed. A key component of the consent decrees was a requirement to conduct emissions tests for volatile organic compounds (VOCs) with the goal of improving the characterization and control of emissions. The conventional VOC stack test method was thought to underquantify total VOC emissions from ethanol plants. A hybrid test method was also developed that involved quantification of individual VOC species. The resulting database of total and speciated VOC emissions from 10 fuel ethanol plants is relatively small, but it is the most extensive to date and has been used to develop and gauge compliance with permit limits and to estimate health risks in Minnesota. Emissions were highly variable among facilities and emissions units. In addition to the variability, the small number of samples and the presence of many values below detection limits complicate the analysis of the data. To account for these issues, a nested bootstrap procedure on the Kaplan Meier method was used to calculate means and upper confidence limits. In general, the fermentation scrubbers and fluid bed coolers emitted the largest mass of VOC emissions. Across most facilities and emissions units ethanol was the pollutant emitted at the highest rate. Acetaldehyde, acetic acid, and ethyl acetate were also important emissions from some units. Emissions of total VOCs, ethanol, and some other species appeared to be a function of the beer feed rate, although the relationship was not reliable enough to develop a production rate-based emissions factor. PMID- 17912929 TI - A laboratory study of sediment and contaminant release during gas ebullition. AB - Significant quantities of gas are generated from labile organic matter in contaminated sediments. The implications for the gas generation and subsequent release of contaminants from sediments are unknown but may include enhanced direct transport such as pore water advection and diffusion. The behavior of gas in sediments and the resulting migration of a polyaromatic hydrocarbon, viz phenanthrene, were investigated in an experimental system with methane injection at the base of a sediment column. Hexane above the overlying water layer was used to trap any phenanthrene migrating out of the sediment layer. The rate of suspension of solid particulate matter from the sediment bed into the overlying water layer was also monitored. The experiments indicated that significant amounts of both solid particulate matter and contaminant can be released from a sediment bed by gas movement with the amount of release related to the volume of gas released. The effective mass transfer coefficient of gas bubble-facilitated contaminant release was estimated under field conditions, being around three orders of magnitude smaller than that of bioturbation. A thin sand-capping layer (2 cm) was found to dramatically reduce the amount of contaminant or particles released with the gas because it could prevent or at least reduce sediment suspension. Based on the experimental observations, gas bubble-facilitated contaminant transport pathways for both uncapped and capped systems were proposed. Sediment cores were sliced to obtain phenanthrene concentration. X-ray computed tomography (CT) was used to investigate the void space distribution in the sediment penetrated by gas bubbles. The results showed that gas bubble migration could redistribute the sediment void spaces and may facilitate pore water circulation in the sediment. PMID- 17912930 TI - Mathematical model for photocatalytic destruction of organic contaminants in air. AB - Photocatalytic oxidation (PCO) was investigated in a bench-scale reactor for the abatement of two airborne organic contaminants: toluene and ethanol. A mathematical model that includes the impacts of light intensity, initial contaminant concentration, catalyst thickness, and relative humidity (RH) on the degradation of organic contaminants in a photocatalytic reactor was developed to describe this process. The commercially available catalyst Degussa-PtTiO2 was selected to compare with the MTU-PtTiO2-350 catalyst, which was synthesized by the sol-gel process, platinized, and calcined at 350 degrees C. For toluene removal using the MTU-PtTiO2-350 catalyst, the degradation rate increased with increases in light intensity from 0.2 to 2.2 mW/cm2 and in catalyst thickness from 0.00037 to 0.00361 cm. However, further increases in light intensity and catalyst thickness had only slight effect on the toluene degradation rate. Increasing the initial concentration from 6.29 to 127.9 microg/L and the RH from 10 to 85% resulted in decreases in the toluene degradation rate. For ethanol removal using the MTU-PtTiO2-350 catalyst, the degradation rate increased more rapidly with an increase in RH from 17 to 56%; the RH had little effect on the ethanol degradation rate while it further increased from 56% to 82%. We discuss applicability of the model to estimate the influence of process variables and to evaluate photocatalyst performance. PMID- 17912931 TI - Source apportionment of fine particulate matter in the southeastern United States. AB - Particulate matter (PM) less than 2.5 microm in size (PM2.5) source apportionment by chemical mass balance receptor modeling was performed to enhance regional characterization of source impacts in the southeastern United States. Secondary particles, such as NH4HSO4, (NH4)2SO4, NH4NO3, and secondary organic carbon (OC) (SOC), formed by atmospheric photochemical reactions, contribute the majority (>50%) of ambient PM2.5 with strong seasonality. Source apportionment results indicate that motor vehicle and biomass burning are the two main primary sources in the southeast, showing relatively more motor vehicle source impacts rather than biomass burning source impacts in populated urban areas and vice versa in less urbanized areas. Spatial distributions of primary source impacts show that each primary source has distinctively different spatial source impacts. Results also find impacts from shipping activities along the coast. Spatiotemporal correlations indicate that secondary particles are more regionally distributed, as are biomass burning and dust, whereas impacts of other primary sources are more local. PMID- 17912932 TI - Histopathological study of epithelial flaps in epi-LASIK. PMID- 17912933 TI - Prospective randomized study of clinical performance of 3 aspheric and 2 spherical intraocular lenses in 250 eyes. AB - PURPOSE: To compare the quality of vision with aspheric and spherical intraocular lenses (IOLs) in pseudophakic patients. METHODS: This prospective, comparative, randomized study included 250 eyes of 125 patients with bilateral cataracts. Patients were randomly assigned to receive either IOLs with a spherical biconvex optic (Acrysof SN6OAT [Alcon] or Sensar AR40e [Advanced Medical Optics, AMO]) or IOLs with an aspheric optic (Acrysof IQ SN6OWF [Alcon], Tecnis Z9000 [AMO], or Sofport L161AO [Bausch & Lomb]). Ophthalmologic examination including best spectacle-corrected visual acuity, pupil size, ocular dominance investigation, contrast sensitivity under mesopic and photopic conditions, and wavefront analysis was performed 2 months postoperatively. RESULTS: Aspheric IOLs showed better contrast sensitivity compared to spherical IOLs at spatial frequencies of 6, 12, and 18 cycles per degree (cpd) under photopic conditions and at all spatial frequencies under mesopic conditions. There was no significant difference among the three aspheric IOLs at all spatial frequencies under either photopic or mesopic conditions. Mean total spherical aberration was statistically lower in dominant eyes with aspheric IOLs (0.05 +/- 0.06, 0.11 +/- 0.1, and 0.19 +/- 0.08 pm for the Tecnis Z9000, Acrysof IQ SN6OWF, and Sofport L161AO, respectively) compared with eyes with spherical IOLs (0.62 +/- 0.24 and 0.46 +/- 0.19 microm for the Acrysof SN6OAT and Sensar AR40e, respectively) for a 5-mm pupil diameter. CONCLUSIONS: The aspheric IOLs had less wavefront aberrations and performed better under both photopic and mesopic contrast sensitivity compared to the spherical IOLs. These findings confirm it is possible to improve the optical performance of IOLs by modifying the surfaces. PMID- 17912934 TI - Standardized analyses of correction of astigmatism with the visian toric phakic implantable collamer lens. AB - PURPOSE: To demonstrate the methodology described by the American National Standards Institute (ANSI) Standard Subcommittee on Astigmatism Analysis reporting a standardized method of presenting astigmatism data by laser systems that reshape the cornea also applies to toric phakic intraocular lens (IOL) data. METHODS: The U.S. Food and Drug Administration trial of the Visian Toric Implantable Collamer Lens comprised 210 eyes of 124 study patients. The mean preoperative manifest refraction spherical equivalent for the study cohort was 9.36 +/- 2.66 diopters (D) (range: -19.50 to -2.38 D). The mean preoperative manifest refractive cylinder was 1.94 +/- 0.84 D (range: 1.00 to 4.00 D). RESULTS: Data were arrayed using the 11 tables and 4 figures suggested in the Subcommittee paper. Interpretation of the data from a toric phakic IOL study was similar to that of laser refractive procedures. The addition of a table of manifest refractive cylinder (in the spectacle plane) over time, which provides a direct clinical measure of clinical efficacy, is suggested. Mean postoperative manifest refractive cylinder was 0.51 +/- 0.48 D (range: 0 to 3.00 D) with 65.6% of eyes having < or = 0.50 D of refractive cylinder at 12 months. A table of observed stability of the toric phakic IOL orientation within the eye obtained by slit-lamp examination also is recommended, which is a useful table not applicable in laser refractive procedures. CONCLUSIONS: The data presented demonstrate that the methodology developed by the ANSI Standard Subcommittee on Astigmatism Analysis, with some minor additions, accurately characterizes the efficacy of toric phakic IOLs in treating astigmatism. PMID- 17912935 TI - Comparison of optical low coherence reflectometry and ultrasound pachymetry in the measurement of central corneal thickness before and after photorefractive keratectomy. AB - PURPOSE: To compare an ultrasound pachymeter with an optical low coherence reflectometry (OLCR) pachymeter for measuring pre- and postoperative central corneal thickness of patients undergoing photorefractive keratectomy (PRK). METHODS: In a prospective, noncomparative, interventional study, 48 myopic eyes (mean manifest refractive spherical equivalent: -4.93 +/- 2.93 diopters [D]) of 30 healthy patients underwent PRK. Pre- and postoperative central corneal thickness was measured by two examiners (E1, E2) with an ultrasound pachymeter and an OLCR pachymeter. Agreement and inter-rater repeatability were determined using the comparison method described by Bland and Altman. RESULTS: The limits of agreement between the two devices ranged from 17.8 microm (E1) to 20.5 microm (E2) preoperatively and from 22.4 microm (E1) to 16.9 microm (E2) postoperatively. The coefficient of inter-rater repeatability ranged from 9.1 microm (ultrasound pachymeter) to 5.4 microm (OLCR pachymeter) preoperatively and from 7.1 microm (ultrasound pachymeter) to 4.7 microm (OLCR pachymeter) postoperatively. CONCLUSIONS: The OLCR pachymeter seems to show better pre- and postoperative repeatability compared to the ultrasound pachymeter. The agreement between the two devices should be considered acceptable for clinical practice. Photorefractive keratectomy did not affect the postoperative agreement and repeatability of the pachymeters. PMID- 17912936 TI - Femtosecond laser and microkeratome corneal flaps: comparison of stromal wound healing and inflammation. AB - PURPOSE: To examine early postoperative wound healing in rabbit corneas that had LASIK flaps formed with three different models (15 KHz, 30 KhZ, and 60 KHz) of a femtosecond laser compared with flaps formed with a microkeratome. METHODS: Thirty-nine rabbit eyes were randomized to receive either no surgery or corneal flaps formed with one of the lasers or the microkeratome. Sixteen eyes also had lamellar cuts with no side cuts with the 30 KHz laser. Animals were sacrificed and corneas processed as frozen sections or fixed for transmission electron microscopy. Frozen sections were evaluated with the TUNEL assay to detect apoptosis, immunocytochemistry for Ki67 to detect cell mitosis, and immunocytochemistry for CD11b to detect mononuclear cells. RESULTS: Rabbit corneas that had flaps formed with the 15 KHz laser had significantly more stromal cell death, greater stromal cell proliferation, and greater monocyte influx in the central and peripheral comea at 24 hours after surgery than corneas that had flaps formed with the 30 KHz or 60 KHz laser or the microkeratome. Results of the 60 KHz laser and microkeratome were not significantly different for any of the parameters at 24 hours, except for mitotic stromal cells at the flap margin. Transmission electron microscopy revealed that the primary mode of stromal cell death at 24 hours after laser ablation was necrosis. CONCLUSIONS: Stromal cell necrosis associated with femtosecond laser flap formation likely contributes to greater inflammation after LASIK performed with the femtosecond laser, especially with higher energy levels that result in greater keratocyte cell death. PMID- 17912937 TI - Accuracy of Orbscan II in the assessment of posterior curvature in patients with myopic LASIK. AB - PURPOSE: To evaluate the accuracy of Orbscan II measurements in assessing posterior corneal curvature in patients undergoing myopic LASIK. METHODS: Using the Orbscan II, posterior corneal curvature was assessed pre- and postoperatively in 304 eyes that underwent myopic LASIK. The radius of curvature and corneal refractive power in diopters (D) were compared using the paired sample t test. RESULTS: The mean pre- and postoperative radius of posterior corneal curvature were 6.49 +/- 0.26 mm and 6.35 +/- 0.30 mm, respectively. Mean pre- and postoperative posterior corneal power were -6.17 +/- 0.25 D and -6.32 +/- 0.30 D, respectively, and the difference (0.14 +/- 0.14 D) was statistically significant (P < .01). CONCLUSIONS: Although the derived value for the power of the postoperative LASIK posterior corneal surface is overestimated using the Orbscan II, this small difference may not be clinically important. Orbscan II measurements can therefore be used (with caution) to measure posterior corneal curvature in patients with myopic LASIK for the assessment of intraocular lens power based on the Gaussian optics formula. PMID- 17912938 TI - Topographic customized photorefractive keratectomy for regular and irregular astigmatism after penetrating keratoplasty using the LIGI CIPTA/LaserSight platform. AB - PURPOSE: To evaluate the use of a software ablation program (Corneal Interactive Programmed Topographic Ablation [CIPTA]) that provides customized photorefractive keratectomy (PRK) to correct astigmatism after keratoplasty. METHODS: In this prospective, noncomparative, consecutive case series, 44 eyes underwent CIPTA for correction of astigmatism after penetrating keratoplasty. Eighteen eyes were treated for regular astigmatism and 26 eyes were treated for irregular astigmatism after penetrating keratoplasty. Orbscan II topography (Bausch & Lomb) and a flying-spot laser (LaserScan 2000; LaserSight) were used. Epithelial debridement with alcohol was performed before PRK in 16 eyes and transepithelial PRK was performed in 28 eyes. Mean target-induced astigmatism was 8.19 +/- 2.68 diopters (D) and 7.68 +/- 4.50 D in the regular and irregular astigmatism groups, respectively. RESULTS: Mean follow-up was 25.4 +/- 13 months. At last postoperative follow-up, 13 (72.2%) and 18 (69.2%) eyes in the regular and irregular astigmatism groups, respectively, had uncorrected visual acuity (UCVA) better than 20/40. Four (22.2%) and 8 (30.7%) eyes in the regular and irregular astigmatism groups, respectively, had UCVA of 20/20. Fourteen (77.7%) and 18 (69.2%) eyes in the regular and irregular astigmatism groups, respectively, were within 1.00 D of attempted correction in spherical equivalent manifest refraction. No eye lost Snellen lines of best spectacle-corrected visual acuity. Mean surgically induced astigmatism was 7.66 +/- 2.70 D and 6.99 +/- 3.80 D for the regular and irregular astigmatism groups, respectively. Index of success of astigmatic correction was 0.138 and 0.137 for the regular and irregular astigmatism groups, respectively. Haze developed in three eyes. CONCLUSIONS: Topography-driven PRK using CIPTA software is a suitable solution for correcting regular and irregular astigmatism after penetrating keratoplasty. PMID- 17912939 TI - LASEK and photorefractive keratectomy for myopia: clinical and confocal microscopy comparison. AB - PURPOSE: To compare postoperative visual acuity and corneal morphology after laser epithelial keratomileusis (LASEK) versus photorefractive keratectomy (PRK) in the correction of low to moderate myopia. METHODS: In a double-blind, randomized clinical trial, 50 myopic patients (mean: -4.5 +/- 1.35 diopters) were randomized to receive LASEK in one eye and PRK in the fellow eye. No mitomycin C eye drops were used in this study. Patients were observed daily for 4 days, then at 1 month and every 3 months up to 1 year. Uncorrected and best-corrected visual acuity (UCVA and BSCVA), manifest refraction, corneal epithelium healing time, postoperative pain, and corneal haze were evaluated. Corneal wound healing was quantified with corneal confocal microscopy. RESULTS: Refractive error, UCVA, and BSCVA were not statistically different between eyes treated with LASEK and PRK. Corneal epithelium healing time was 2.52 +/- 0.99 days in the eyes treated with PRK and 2.29 +/- 0.52 days in the eyes treated with LASEK (P=.22). The postoperative pain score was 2.17 +/- 0.87 in the eyes treated with PRK and 2.62 +/- 0.60 (P=.02) in the eyes treated with LASEK. Corneal confocal microscopy showed fewer stromal activated keratocytes and less extracellular matrix deposition in the eyes treated with LASEK than in the eyes treated with PRK at 1 month postoperatively (P=.003). CONCLUSIONS: LASEK is an effective and safe procedure for low to moderate myopia, but it seems more painful until full corneal reepithelization. In the early postoperative period, the corneal wound healing process is significantly less intense in eyes treated with LASEK than in eyes treated with PRK. The role of LASEK in corneal wound healing modulation remains controversial. PMID- 17912941 TI - Subbasal nerve regeneration after LASEK measured by confocal microscopy. AB - PURPOSE: To determine the relationship between the degree of subbasal nerve regeneration and corneal sensation and tear film functions after LASEK using corneal confocal microscopy. METHODS: This prospective, observational, and longitudinal study included 35 LASEK patients who underwent tear breakup time assessment, phenol red thread test, corneal esthesiometry, and confocal microscopy preoperatively and 1, 3, and 6 months postoperatively. Subbasal nerve images were analyzed to assess nerve regeneration. RESULTS: Postoperatively, tear breakup time decreased significantly and had not returned to the preoperative level by 6 months after surgery. There were no significant differences in the phenol red thread test results before and after LASEK. Central corneal sensation decreased significantly 1 month after LASEK and returned to normal levels 3 months after surgery. Nerve fiber density, nerve branch density, and nerve fiber length and width decreased significantly after LASEK and had not returned to preoperative levels 6 months after surgery. Subbasal nerve fibers retained their vertical orientation after LASEK. There were no significant correlations between the length and density of subbasal nerve fibers and central corneal sensitivity, tear volume, or breakup time after surgery. CONCLUSIONS: Subbasal nerves were injured by LASEK and had not returned to preoperative levels 6 months after surgery. Corneal sensitivity was reduced after LASEK and returned to normal levels 3 months after surgery. PMID- 17912940 TI - Phototherapeutic keratectomy in children. AB - PURPOSE: Children with dense superficial opacities of the cornea are at risk for developing amblyopia. This study evaluated the efficacy of phototherapeutic keratectomy (PTK) in severe cases of anterior stromal scarring. METHODS: Five eyes of five patients, aged 6 to 8 years, were included. Three children suffered from uveitis-associated band keratopathy, one child had anterior corneal stromal scarring due to viral infection, and one child had anterior basement membrane dystrophy. Phototherapeutic keratectomy was performed under general anesthesia using the 200 Hz ALLEGRETTO excimer laser. Optical treatment zone was 7.0 mm (one eye) and 8.0 mm (four eyes), and ablation depths were between 20 and 100 microm. Postoperative treatment consisted of bandage soft contact lens, topical preservative-free antibiotics, steroids, and artificial tears. Part-time occlusion therapy was continued, as prior to PTK. Because of the small cohort, statistical evaluation was not performed. RESULTS: Mean follow-up was 23.4 +/- 13.7 months (range: 10 to 41 months). Surgery and postoperative follow-up were uneventful, and no signs of infection or haze were noted. Preoperative best spectacle-corrected visual acuity was 0.4 to 3.0 logMAR (mean: 1.22 +/- 1.07 logMAR). Best spectacle-corrected visual acuity improved in four eyes and stabilized in one eye due to uveitis recurrence (mean 0.64 +/- 0.65 logMAR). Mean preoperative keratometric values changed from 45.60 +/- 0.90 to 44.90 +/- 3.00 diopters postoperatively. CONCLUSIONS: Dense superficial corneal opacity in children may be successfully treated with PTK. Phototherapeutic keratectomy was performed to increase corneal transparency and corneal surface smoothing thereby avoiding amblyopia. PMID- 17912942 TI - Corneal light shield as a delivery system for standardized application of mitomycin C in excimer surface ablation. AB - PURPOSE: To develop a simple, reproducible method of applying intraoperative mitomycin C (MMC) in excimer surface ablation surgery. METHODS: A two-part protocol was developed to study several properties of corneal light shields. Part A tested the amount of MMC (0.2 mg/mL) absorbed, expansion dimension attained, and amount released to filter paper. Part B examined in vitro release of MMC to a photorefractive keratectomy (PRK)-treated enucleated pig eye. RESULTS: Mean MMC absorbed by corneal light shields was 63.7 +/- 5.0 mg (range: 57.4 to 72.8 mg); coefficient of variability was 7.8%. Expansion diameter and thickness were constant at 9.0 mm and 1.1 mm, respectively. Mean solution released to filter paper was 55.4 +/- 3.9 mg (range: 51.3 to 63.5 mg); coefficient of variability was 7.1%. Mean solution transferred to the PRK-treated pig eye was 2.3 +/- 0.7 mg (range: 1.1 to 3.5 mg); coefficient of variability was 33%. CONCLUSIONS: Mitomycin C contact surface area, amount absorbed, and amount released by each corneal light shield were reproducible. Uniform dimensions theoretically provide uniform distribution of MMC. This method may allow standardization of intraoperative MMC application in excimer surface ablation. PMID- 17912943 TI - Idiopathic recurrence of diffuse lamellar keratitis after LASIK. AB - PURPOSE: To report a case of late recurrence of bilateral diffuse lamellar keratitis (DLK) after LASIK. METHODS: A 39-year-old woman presented in the early postoperative period with bilateral DLK after hyperopic LASIK and was treated with topical steroids. One year after and with no obvious cause (idiopathic), recurrence of the same stage (stage III) of disease was observed. RESULTS: Slit lamp examination revealed diffuse, multifocal, and granular haze in the interface. The microbiology culture was negative. Confocal microscopy demonstrated multiple activated keratocytes, debris, and inflammatory cells adjacent to the flap interface. After intense treatment with topical corticosteroids, DLK resolved and corneal transparency was achieved with complete restoration of visual acuity. CONCLUSIONS: Diffuse lamellar keratitis may recur in LASIK patients with previous episodes without an obvious cause (idiopathic). Early diagnosis and treatment with topical corticosteroids can achieve complete resolution without visual loss--even in advanced stages of DLK. PMID- 17912944 TI - Small spot phototherapeutic keratectomy for recurrent corneal erosion. AB - PURPOSE: To describe small spot phototherapeutic keratectomy (PTK) using the NIDEK EC-5000 excimer laser for the treatment of recurrent corneal erosion. METHODS: This retrospective study analyzed patients with recurrent comeal erosion who were treated using small spot PTK. An electronic medical records database was used to retrieve patient data. RESULTS: Ten consecutive eyes in nine patients were evaluated after small spot PTK. Eight (80%) recovered without further erosion. CONCLUSIONS: Small spot PTK is a safe and effective alternative to traditional PTK and to corneal micropuncture in the treatment of recurrent corneal erosion. PMID- 17912945 TI - Effect of mitomycin C on the corneal endothelium when used for corneal subepithelial haze prophylaxis following photorefractive keratectomy. AB - PURPOSE: To evaluate the potential effect of topical mitomycin C (MMC) on the corneal endothelium of myopic patients undergoing photorefractive keratectomy (PRK). METHODS: Sixteen eyes with a planned ablation depth >75 microm underwent PRK followed by 0.02% MMC applied for 12 seconds using a methylcellulose sponge. Endothelial specular microscopy was performed with the Keeler-Konan specular photomicroscope in 16 eyes before and at least 1 year after surgery. Mean follow up was 18 months (range: 12 to 24 months). Mean cell density, coefficient of variation of mean cell area, and percentage of hexagonal cells were measured and calculated using computerized morphometric analysis. RESULTS: Mean endothelial cell densities before and after surgery were 2882 +/- 783 cells/mm2 (range: 1511 to 4022 cells/mm2) and 2867 +/- 588 cells/mm2 (range: 1638 to 3881 cells/mm2), respectively (P > .05). Mean coefficient of variation before and after surgery was 0.30 +/- 0.07 (range: 0.23 to 0.49) and 0.26 +/- 0.04 (range: 0.22 to 0.33), respectively (P=.06). Mean percentage of hexagonal cells before and after surgery was 61% +/- 6.8% (range: 47% to 70%) and 66% +/- 6.7% (range: 54% to 75%), respectively. CONCLUSIONS: Administration of MMC for haze prophylaxis following PRK did not have a significant effect on quantitative endothelial cell density or qualitative morphometric parameters in this study. PMID- 17912946 TI - Aureobasidium pullulans fungal keratitis following LASEK. AB - PURPOSE: To describe a patient who developed Aureobasidium pullulans keratitis following refractive laser epithelial keratomileusis (LASEK). METHODS: A 52-year old woman was referred to a tertiary care center 1 month after LASEK for treatment of a corneal ulcer that was unresponsive to conventional therapy. Mycology culture and fungal stain identified Aureobasidium as the infectious organism. RESULTS: The infection responded well to treatment with topical natamycin and systemic itraconazole. CONCLUSIONS: Treatment with topical natamycin and systemic itraconazole is effective against Aureobasidium pullulans keratitis. PMID- 17912947 TI - Traumatic flap dislocation 4 years after LASIK due to air bag injury. AB - PURPOSE: To report a patient who developed corneal flap dislocation following air bag injury 48 months after LASIK. METHODS: Evaluation by slit-lamp microscopy and fluorescein angiography. RESULTS: A 29-year-old man was treated after air bag injury that occurred 48 months after LASIK. Examination revealed corneal flap dislocation, with severe folds and flap edema. Preoperative visual acuity was finger counting at 1 m. Visual acuity was 20/400 24 hours after repositioning the corneal flap. Retinal angiography revealed Berlin macular edema, which was injected with periocular steroids. Five days after injection, visual acuity remained 20/400, but improved to 20/40 1 month after injection. CONCLUSIONS: Significant trauma can dislocate a corneal flap many months after surgery. PMID- 17912948 TI - Autologous transplantation of muscle-derived CD133+ stem cells in Duchenne muscle patients. AB - Duchenne muscular dystrophy (DMD) is a lethal X-linked recessive muscle disease due to defect on the gene encoding dystrophin. The lack of a functional dystrophin in muscles results in the fragility of the muscle fiber membrane with progressive muscle weakness and premature death. There is no cure for DMD and current treatment options focus primarily on respiratory assistance, comfort care, and delaying the loss of ambulation. Recent works support the idea that stem cells can contribute to muscle repair as well as to replenishment of the satellite cell pool. Here we tested the safety of autologous transplantation of muscle-derived CD133+ cells in eight boys with Duchenne muscular dystrophy in a 7 month, double-blind phase I clinical trial. Stem cell safety was tested by measuring muscle strength and evaluating muscle structures with MRI and histological analysis. Timed cardiac and pulmonary function tests were secondary outcome measures. No local or systemic side effects were observed in all treated DMD patients. Treated patients had an increased ratio of capillary per muscle fibers with a switch from slow to fast myosin-positive myofibers. PMID- 17912949 TI - Ex vivo expansion and transplantation of hematopoietic stem/progenitor cells supported by mesenchymal stem cells from human umbilical cord blood. AB - Human mesenchymal stem cells (MSCs) are multipotential and are detected in bone marrow (BM), adipose tissue, placenta, and umbilical cord blood (UCB). In this study, we examined the ability of UCB-derived MSCs (UCB-MSCs) to support ex vivo expansion of hematopoietic stem/progenitor cells (HSPCs) from UCB and the engraftment of expanded HSPCs in NOD/SCID mice. The result showed that UCB-MSCs supported the proliferation and differentiation of CD34+ cells in vitro. The number of expanded total nucleated cells (TNCs) in MSC-based culture was twofold higher than cultures without MSC (control cultures). UCB-MSCs increased the expansion capabilities of CD34+ cells, long-term culture-initiating cells (LTC ICs), granulocyte-macrophage colony-forming cells (GM-CFCs), and high proliferative potential colony-forming cells (HPP-CFCs) compared to control cultures. The expanded HSPCs were transplanted into lethally irradiated NOD/SCID mice to assess the effects of expanded cells on hematopoietic recovery. The number of white blood cells (WBCs) in the peripheral blood of mice transplanted with expanded cells from both the MSC-based and control cultures returned to pretreatment levels at day 25 posttransplant and then decreased. The WBC levels returned to pretreatment levels again at days 45-55 posttransplant. The level of human CD45+ cell engraftment in primary recipients transplanted with expanded cells from the MSC-based cultures was significantly higher than recipients transplanted with cells from the control cultures. Serial transplantation demonstrated that the expanded cells could establish long-term engraftment of hematopoietic cells. UCB-MSCs similar to those derived from adult bone marrow may provide novel targets for cellular and gene therapy. PMID- 17912950 TI - A closed system for islet isolation and purification using the COBE2991 cell processor may reduce the need of clean room facilities. AB - During the isolation of human islets of Langerhans the digest has repeated direct contact with the ambient atmosphere. In order to fulfill GMP requirements in clinical applications, the entire cell preparation must be performed in clean room facilities. We hypothesized that the use of a closed system, which avoids the direct exposure of tissue to the atmosphere, would significantly ease the preparation procedure. To avoid the direct atmosphere exposure we tested a modification of the isolation and purification process by performing all islet preparation steps in a closed system. In this study we compared the isolation outcome of the traditional open preparation technique with the new closed system. Pancreata from 6-month-old hybrid pigs were procured in the local slaughterhouse. After digestion/filtration the digest was cooled, collected, and concentrated in centrifugation containers and purified thereafter in the COBE2991 by top loading (control). In the control group 502 +/- 253 IEQ per gram pancreas were purified. The total preparation time amounted to 12 h. In the closed system the digest was cooled and directly pumped into the COBE2991 for centrifugation followed by supernatant expelling. Bag filling, centrifugation, and expelling were repeated several times. Islets in pellet form were then purified by adding a gradient (bottom loading). Using this closed system 1098 +/- 489 IEQ per gram pancreas were purified with a total cell viability of 67 +/- 10% and a beta-cell viability of 41 +/- 13%. The total preparation time reduced to 6 h. After 24 h of cell culture the viability of beta-cells was still 56 +/- 10% and was only reduced after the addition of proapoptotic IL-1 and TNF-alpha to 40 +/- 4%, indicating that freshly isolated islets are not apoptotic. In conclusion, the closed system preparation is much faster, more effective, and less expensive than the traditional islet preparation. The closed system may be applicable for human islets preparations to restrict the need of clean room facilities for islet preparations to a minimum and may open the way for islet preparations without clean room demand. PMID- 17912952 TI - Encapsulation of porcine islets permits extended culture time and insulin independence in spontaneously diabetic BB rats. AB - The ability to culture porcine islets for extended times allows for both their functional assessment and the assurance of their microbiological safety prior to transplantation. We have previously shown that agarose-encapsulated porcine islets can be cultured for at least 24 weeks. In the current study, porcine islet agarose macrobeads cultured for up to 67 weeks were assessed for their ability to restore normoglycemia, respond to an intraperitoneal glucose challenge, maintain spontaneously diabetic BB rats free of insulin therapy for more than 6 months, and for their biocompatibility. Porcine islets were encapsulated in agarose macrobeads and subjected to weekly static perifusion assays for the assessment of insulin production. After in vitro culture for either 9, 40, or 67 weeks, 56-60 macrobeads were transplanted to each spontaneously diabetic BB rat. Transplanted rats were monitored daily for blood glucose levels. Glucose tolerance tests and assessments for porcine C-peptide were conducted at various intervals throughout the study. Normoglycemia (100-200 mg/dl) was initially restored in all islet transplanted rats. Moderate hyperglycemia (200-400 mg/dl) developed at around 30 days posttransplantation and continued throughout the study period of 201-202 days. Importantly, all rats that received encapsulated porcine islets continued to gain weight and were free of exogenous insulin therapy for the entire study. Porcine C-peptide (0.2-0.9 ng/ml) was detected in the serum of islet recipients throughout the study period. No differences were detected between recipient animals receiving islet macrobeads of various ages. These results demonstrate that the encapsulation of porcine islets in agarose macrobeads allows for extended culture periods and is an appropriate strategy for functional and microbiological assessment prior to clinical use. PMID- 17912951 TI - Toward maximizing the success rates of human islet isolation: influence of donor and isolation factors. AB - In order to make islet transplantation a therapeutic option for patients with diabetes there is an urgent need for more efficient islet cell processing to maximize islet recovery. Improved donor management, organ recovery techniques, implementation of more stringent donor criteria, and improved islet cell processing techniques may contribute to enhance organ utilization for transplantation. We have analyzed the effects of donor and islet processing factors on the success rate of human islet cell processing for transplantation performed at a single islet cell processing center. Islet isolation outcomes improved when vasopressors, and in particular pitressin, and steroids were used for the management of multiorgan donors. Higher islet yields were obtained from adult male donors, BMI >25 kg/m2, adequate glycemic control during hospital stay, and when the pancreas was retrieved by a local surgical team. Successful isolations were obtained in 58% of the cases when > or = 4 donor criteria were met, and even higher success rates (69%) were observed when considering > or = 5 criteria. Our data suggest that a sequential, integrated approach is highly desirable to improve the success rate of islet cell processing. PMID- 17912953 TI - Distribution of intraportally implanted microspheres and fluorescent islets in mice. AB - The aim of the study was to evaluate the distribution of intraportally transplanted islets in mice. We initially administered 2000 polystyrene microspheres with a diameter of 50 microm intraportally into normoglycemic C57BL/6 mice. In separate experiments other mice were injected similarly with 300 microspheres each with a diameter of 100 or 200 microm. One week later the animals were killed, and the lungs and livers were removed and divided into lobes. The number of microspheres in each individual liver lobe and in the lungs was counted using a stereomicroscope. In other experiments, athymic C57BL/6 mice were similarly implanted with 250 islets isolated from transgenic mice expressing the enhanced yellow fluorescent protein in the islet cells. The distribution of microspheres and islets was independent of size, and fairly homogenous within the liver, with the exception of the caudate lobe, which contained fewer microspheres and islets, respectively. Approximately one third of all microspheres and islets were present as aggregates. Eighty-five to 90% of the implanted microspheres were identified in the liver sections, whereas 60-65% of the implanted islets were recovered. Aggregates or single fluorescent cells were observed in the liver of islet-implanted mice. We conclude that islets and microspheres implanted into the liver distribute fairly homogenously and quite a few of them exist as aggregates or, with respect to islets, as fragments. PMID- 17912954 TI - Hepatocyte transplantation for glycogen storage disease type Ib. AB - Glycogen storage disease type I (GSD-I) is a group of autosomal recessive disorders with an incidence of 1 in 100,000. The two major subtypes are GSD-Ia, caused by a deficiency of glucose-6-phosphatase (G6Pase), and GSD-Ib, caused by a deficiency of glucose-6-phosphate transporter (G6PT). We report that a substantial improvement was achieved following several infusions of hepatocytes in a patient with GSD-Ib. Hepatocytes were isolated from the unused cadaveric whole livers of two donors. At the first transplantation, approximately 2 x 10(9) cells (2% of the estimated recipient's total hepatocytes) were infused. Seven days later 1 x 10(9) (1% of liver mass) cryopreserved hepatocytes from the same donor were infused, and an additional 3 x 10(9) (3% of liver mass) cells from the second donor were infused 1 month after the second transplantation. After the hepatocyte transplantation, the patient showed no hypoglycemic symptoms despite the discontinuation of cornstarch meals. Liver biopsies on posttransplantation days 20 and 250 showed a normal level of glucose-6-phosphatase activity in presolubilization assay that was very low before transplantation. This was the first and successful clinical hepatocyte transplantation in Korea. In this study, hepatocyte transplantation allowed a normal diet in a patient with GSD-Ib, with substantial improvement in their quality of life. Hepatocyte transplantation might be an alternative to liver transplantation and dietary therapy in GSD-Ib. PMID- 17912955 TI - Cryopreservation-induced nonattachment of human hepatocytes: role of adhesion molecules. AB - Good quality cryopreserved human hepatocytes are becoming an important source for clinical hepatocyte transplantation. However, the process of cryopreservation leads to both structural and functional impairment of hepatocytes. The aim of this study was to investigate the mechanisms of cryopreservation-induced nonattachment in human hepatocytes. Hepatocytes were cryopreserved after isolation from unused donor liver tissue. Cell attachment to collagen-coated plates was measured. A cDNA gene array system for 96 cell adhesion-related molecules was used to determine mRNA expression in fresh and cryopreserved hepatocytes. Two cell adhesion molecule proteins were investigated further: beta1 integrin, a cell-matrix adhesion molecule, and E-cadherin, a cell-cell adhesion molecule. Attachment efficiency was significantly decreased after cryopreservation of human hepatocytes. Twenty-two genes were downregulated after cryopreservation including integrins, cadherins, catenins, and matrix metalloproteinases (MMPs). Beta1-Integrin gene and protein expression were significantly decreased in cultured cryopreserved hepatocytes compared to fresh hepatocytes. There was a significant correlation between loss of beta1-integrin and attachment in cryopreserved cells. Degradation of E-cadherin was increased in cryopreserved hepatocytes. The process of cryopreservation leads to downregulation of cell adhesion molecules at the gene and the cellular level. New cryopreservation protocols are needed to prevent these effects on cell attachment. PMID- 17912956 TI - Culture of keratinocytes for transplantation without the need of feeder layer cells. AB - Patients with large burn wounds have a limited amount of healthy donor skin. An alternative for the autologous skin graft is transplantation with autologous keratinocytes. Conventionally, the keratinocytes are cultured with mouse feeder layer cells in medium containing fetal calf serum (FCS) to obtain sufficient numbers of cells. These xenobiotic materials can be a potential risk for the patient. The aim of the present study was to investigate if keratinocytes could be expanded in culture without the need of a feeder layer and FCS. Keratinocytes were cultured on tissue culture plastic with or without collagen type IV coating in medium containing Ultroser G (serum substitute) and keratinocyte growth factor (KGF). An in vitro skin equivalent model was used to examine the capacity of these cells to form an epidermis. Keratinocytes in different passages (P2, P4, and P6) and freshly isolated cells were studied. Keratinocytes grown on collagen type IV were able to form an epidermis at higher passage numbers than cells grown in the absence of collagen type IV (P4 and P2, respectively). In both cases the reconstructed epidermis showed an increased expression of Ki-67, SKALP, involucrin, and keratin 17 compared to normal skin. Only 50,000 keratinocytes grown on collagen type IV in P4 were needed to form 1 cm2 epidermis, whereas 150,000 of freshly isolated keratinocytes were necessary. Using this culture technique sufficient numbers of keratinocytes, isolated from 1 cm2 skin, were obtained to cover 400 cm2 of wound surface in 2 weeks. The results show that keratinocytes can be cultured without the need of a fibroblast feeder layer and FCS and that these cells are still able to create a fully differentiated epidermis. This culture technique can be a valuable tool for the treatment of burn wounds and further development of tissue engineered skin. PMID- 17912957 TI - Inhibition of apoptosis by expression of antiapoptotic proteins in recombinant human keratinocytes. AB - The Fas ligand/Fas interaction plays an important role in the regulation of immune responses. Allografted cells undergo Fas-mediated apoptosis induced by CD8+ T cells. Our objective was to prevent human keratinocytes from immunologically induced apoptosis. We focused on three proteins with inhibitory function on Fas-mediated apoptosis. Human keratinocytes were transfected with either Flip, Faim, or Lifeguard (LFG). The treatment proved to be practicable and efficient. The recombinant keratinocytes with expression of our target proteins were cocultured with CD8+ T cells and the apoptotic activity was then evaluated. Activation of caspase-8 was detectable in control but not in the recombinant cells. Quantitative analysis revealed significant induction of T-cell-induced apoptosis in nontransfected keratinocytes (p = 0.04, n = 12) but not in Flip (p = 0.66), Faim (p = 0.42), or LFG (p = 0.44) expressing cells. Our results suggest that heterotopic expression of antiapoptotic proteins can induce the resistance of keratinocytes to a major mechanism of rejection. PMID- 17912958 TI - Titanium dental implant surface micromorphology optimization. AB - The purpose of this investigation was to create an acid-etched implant surface that is similar to that created by sandblasting combined with acid etching and to compare it with the surfaces of various commercially available screw-type implants. Titanium grade 5 disks were machined in preparation for acid etching. Tests were carried out using different acids and combinations of them with varying time exposures. All etched surfaces were scanned with an electron microscope, and digital images were created for visual evaluation and description of the surfaces. The etched surfaces were evaluated for surface morphology (combination of microroughness and waviness); the surface most like the sandblasted/acid-etched surface was best obtained with a combination of sulfuric and hydrochloric acids. The etched titanium disks were fixed in acrylic resin (2 were cut and polished and 2 were scored and fractured) and the surface profile was examined. In the second part of the investigation, 28 screw-shaped implants that were manufactured from commercially available titanium grade 5 were selected and divided into 2 groups: 3 implants were used as controls (machined surface), and 25 implants were processed using the preferred etching method determined in the first part of the investigation. Magnifications of 27, 200, and 2000 were used to analyze the first 2 consecutive crests of threads, flanks, and root of threads of each implant with the treated surface. A 3-dimensional optical interferometer was used to characterize the surface roughness of both control and test groups. Three screws were selected from each group and measured at 9 sites: 3 measurements each on the crest, root, and flank of the threads. To describe the surface roughness in numbers, the following parameters were used: the average height deviation (Sa), the developed interfacial area ratio (Sdr), the fastest decay autocorrelation length (Sal), and the density of summits (Sds). In addition, in a third experiment, the surfaces of 5 commercially available screw type implants and the experimental ones were analyzed and compared. It was concluded that the new experimental acid-etched titanium surface had the features of a roughened titanium surface, with glossily microroughness and large waviness. In general, the experimental surface was significantly rougher than the selected commercially available implants and similar to a sandblasted/acid-etched surface (top Sa: 2.08 +/- 0.36 microm, Sdr: 1.34 +/- 0.3 microm, valleys: 1.16 +/- 0.1 microm and 0.68 +/- 0.1 microm, flanks: 2.24 +/- 0.8 microm and 1.27 +/- 0.1 microm, respectively). PMID- 17912959 TI - A survey of radiographic measurement estimation in assessment of dental implant length. AB - The aim of this study is to survey radiographic measurement estimation in the assessment of dental implant length according to dentists' confidence. A 19-point questionnaire with closed-ended questions was used by two graduate students to interview 69 dentists during a dental implant meeting. Included were 12 questions related to over- and underestimation in three radiographic modalities: panoramic (P), conventional tomography (T), and computerized tomography (CT). The database was analyzed by Epi-Info 6.04 software and the values from two radiographic modalities, P and T, were compared using a chi2 test. The results showed that 38.24% of the dentists' confidence was in the overestimation of measurements in P, 30.56% in T, and 0% in CT. On the other hand, considering the underestimated measurements, the percentages were 47.06% in P, 33.33% in T, and 1.92% in CT. The frequency of under- and overestimation were statistically significant (chi2 = 6.32; P = .0425) between P and T. CT was the radiographic modality with higher measurement precision according to dentists' confidence. In conclusion, the interviewed dentists felt that CT was the best radiographic modality when considering the measurement estimation precision in preoperative dental implant assessment. PMID- 17912960 TI - Dermal fillers for facial soft tissue augmentation. AB - Nowadays, patients are demanding not only enhancement to their dental (micro) esthetics, but also their overall facial (macro) esthetics. Soft tissue augmentation via dermal filling agents may be used to correct facial defects such as wrinkles caused by age, gravity, and trauma; thin lips; asymmetrical facial appearances; buccal fold depressions; and others. This article will review the pathogenesis of facial wrinkles, history, techniques, materials, complications, and clinical controversies regarding dermal fillers for soft tissue augmentation. PMID- 17912962 TI - Implant zones of the jaws: implant location and related success rate. AB - The article demonstrates the factors of importance in the early and late failures of dental implants based on literature review. An implant location is one of many factors that can influence a success or failure of dental implants. The author identifies and describe four alveolar jaw regions--functional implant zones--with unique characteristics of anatomy, blood supply, pattern of bone resorption, bone quality and quantity, need for bone grafting and other supplemental surgical procedures, and a location-related implant success rate. The article discusses predisposing factors that can lead to early implant failures in different jaw zones. An implant location is investigated as one of these factors. A prior history of trauma to premaxillary region is described in the context of implant success in anterior maxilla. This zone is being referred by the author as the "traumatic zone." The challenges of mandibular posterior implant reconstruction are presented in the context of blood supply to the mandible. A deficiency of vascularization in this region, especially in elderly and edentulous patients, lead the author to refer to this zone as the "ischemic zone." The concept of relative ischemia of the posterior mandible that can develop with age and tooth loss is discussed. A thorough understanding of specifics of each functional implant zone should help to improve successes and prevent failures of dental implants. PMID- 17912961 TI - Minimally invasive guided bone regeneration. AB - Guided bone regeneration (GBR) is indicated when there is a volume deficiency of the residual ridge that prohibits implantation or optimal implant installation for esthetic and functional needs. Our objective is to describe a new minimally invasive GBR and report its clinical application in several patients. A vertical incision is made mesial to the augmentation zone. The periosteum is initially elevated with a miniature chisel, and then through a series of sequential balloon inflations. This yields a tunnel with adequate space for membrane insertion, decortication, and grafting with substitute bone and platelet rich fibrin (PRF) filling. Primary closure is obtained by 2 or 3 simple interrupted sutures. Vertical and horizontal gains were measured on computerized tomography obtained before and 5 to 6 months after the procedure. Eleven patients were treated with this procedure. There were no significant adverse events. The range of vertical gain was 2.4 to 5.1 mm, while horizontal gain measured 1.3 to 3.9 mm. Implants were successfully placed in 6 patients. PMID- 17912963 TI - The prevalence and significance of anatomic variance in the mandibular symphysis: a retrospective study. AB - As computerized tomography (CT) becomes more widespread in implant treatment planning, dentists are bound to encounter more uncharacteristic anatomical conditions in the jaws. This retrospective study examined the prevalence of one such anatomic variance on 78 CT scans of human mandibles. Irregular heterogeneous circular radiopacities with internal radiolucencies were present in a majority (97.4%) of the CT scans examined; however, the phenomenon only appeared pronounced in 28.9% of this group. Bone radiodensity was higher in the regions with the phenomenon as compared with adjacent bone without the phenomenon, which suggested no adverse indications for implant placement. PMID- 17912964 TI - Prospective clinical evaluation of 835 multithreaded tapered screw-vent implants: results after two years of functional loading. AB - Multithreaded tapered screw implants have been used for several years, but lack of clinical documentation about marginal bone stability and survival rates have raised concerns about the design among some clinicians. This study prospectively evaluated the survival rates, success rates, and marginal bone stability of multithreaded tapered screw implants. A total of 835 implants in diameters of 3.7 mm (9%), 4.7 mm (76%), and 6.0 mm (15%) were placed in 328 patients using a single-stage, delayed-loading protocol. The implants were restored with a variety of prostheses and monitored over 2 years of functional loading. Five implants failed and were removed before loading. Cumulative implant survival was 99.4% (n = 835); differences between mandibular (99.0%, n = 408) and maxillary (99.8%, n = 427) implants were not statistically significant (P > .20). Mean marginal bone resorption was 1.66 mm (+/- 0.13 mm). Six implants failed to meet the success criteria by sustaining mesial and distal bone loss below the first implant thread; however, they remained stable and continued functioning without pain or inflammation. Cumulative implant success was 98.6% (n = 835); differences between maxillary (98.6%) and mandibular (98.8%) implants were not statistically significant (P > .20). Success rates by implant diameter were 98.6% (3.7 mm), 98.4% (4.7 mm), and 100% (6 mm). After 2 years of functional loading, survival and success rates for multithreaded tapered implants placed in a nonsubmerged protocol equaled or surpassed those of single-thread, straight-walled implant historical controls. PMID- 17912965 TI - Implant surface analysis and microbiologic evaluation of failed implants retrieved from smokers. AB - The aim of this study was to evaluate the microbiota and surface of failed titanium dental implants from 4 manufacturers. Twelve mobile dental implants were retrieved from 10 smokers after 3 to 10 years of functional loading. Before implant removal, microbial samples were taken and evaluated using polymerase chain reaction. After implant removal, analyses of the failed implant surfaces were performed using scanning electron microscopy and energy-dispersive spectrometer x-ray. Periodontal pathogens such as Aggregactibacter actinomycetemcomitans, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, and Treponema denticola were detected in all implants in different proportions. Surface analysis showed varying degrees of surface roughness between the samples and the presence of proteinaceous material, appearing mainly as dark stains. Foreign carbon, oxygen, sodium, calcium, aluminum, and silicon elements were also found. Although no material-related causes of implant failure were detected, several periodontal pathogens were identified independently of the surface topography or manufacturer. PMID- 17912966 TI - Simulating 3-D lung dynamics using a programmable graphics processing unit. AB - Medical simulations of lung dynamics promise to be effective tools for teaching and training clinical and surgical procedures related to lungs. Their effectiveness may be greatly enhanced when visualized in an augmented reality (AR) environment. However, the computational requirements of AR environments limit the availability of the central processing unit (CPU) for the lung dynamics simulation for different breathing conditions. In this paper, we present a method for computing lung deformations in real time by taking advantage of the programmable graphics processing unit (GPU). This will save the CPU time for other AR-associated tasks such as tracking, communication, and interaction management. An approach for the simulations of the three-dimensional (3-D) lung dynamics using Green's formulation in the case of upright position is taken into consideration. We extend this approach to other orientations as well as the subsequent changes in breathing. Specifically, the proposed extension presents a computational optimization and its implementation in a GPU. Results show that the computational requirements for simulating the deformation of a 3-D lung model are significantly reduced for point-based rendering. PMID- 17912967 TI - A mobile care system with alert mechanism. AB - Hypertension and arrhythmia are chronic diseases, which can be effectively prevented and controlled only if the physiological parameters of the patient are constantly monitored, along with the full support of the health education and professional medical care. In this paper, a role-based intelligent mobile care system with alert mechanism in chronic care environment is proposed and implemented. The roles in our system include patients, physicians, nurses, and healthcare providers. Each of the roles represents a person that uses a mobile device such as a mobile phone to communicate with the server setup in the care center such that he or she can go around without restrictions. For commercial mobile phones with Bluetooth communication capability attached to chronic patients, we have developed physiological signal recognition algorithms that were implemented and built-in in the mobile phone without affecting its original communication functions. It is thus possible to integrate several front-end mobile care devices with Bluetooth communication capability to extract patients' various physiological parameters [such as blood pressure, pulse, saturation of haemoglobin (SpO2), and electrocardiogram (ECG)], to monitor multiple physiological signals without space limit, and to upload important or abnormal physiological information to healthcare center for storage and analysis or transmit the information to physicians and healthcare providers for further processing. Thus, the physiological signal extraction devices only have to deal with signal extraction and wireless transmission. Since they do not have to do signal processing, their form factor can be further reduced to reach the goal of microminiaturization and power saving. An alert management mechanism has been included in back-end healthcare center to initiate various strategies for automatic emergency alerts after receiving emergency messages or after automatically recognizing emergency messages. Within the time intervals in system setting, according to the medical history of a specific patient, our prototype system can inform various healthcare providers in sequence to provide healthcare service with their reply to ensure the accuracy of alert information and the completeness of early warning notification to further improve the healthcare quality. In the end, with the testing results and performance evaluation of our implemented system prototype, we conclude that it is possible to set up a complete intelligent healt care chain with mobile monitoring and healthcare service via the assistance of our system. PMID- 17912968 TI - An interactive framework for personalized computer-assisted neurorehabilitation. AB - This paper presents the implementation of a framework for computer-assisted neurorehabilitation that intends to address the need for more personalized healthcare technologies. This framework called UniTherapy is applied to home neurorehabilitation for individuals with stroke-induced disability. It supports interactive upper limb assessment and therapy that makes use of mass-marketed force-reflecting joysticks and wheels, as well as some customized therapeutic devices. A novel service-oriented technical infrastructure is presented, which includes a rich menu of performance assessment capabilities and support features that include telerehabilitation links, protocol design, and data analysis tools. Results are presented that demonstrate its potential as a sensor-based assessment tool. User feedback is summarized. PMID- 17912969 TI - User-centered evaluation of Arizona BioPathway: an information extraction, integration, and visualization system. AB - Explosive growth in biomedical research has made automated information extraction, knowledge integration, and visualization increasingly important and critically needed. The Arizona BioPathway (ABP) system extracts and displays biological regulatory pathway information from the abstracts of journal articles. This study uses relations extracted from more than 200 PubMed abstracts presented in a tabular and graphical user interface with built-in search and aggregation functionality. This paper presents a task-centered assessment of the usefulness and usability of the ABP system focusing on its relation aggregation and visualization functionalities. Results suggest that our graph-based visualization is more efficient in supporting pathway analysis tasks and is perceived as more useful and easier to use as compared to a text-based literature-viewing method. Relation aggregation significantly contributes to knowledge-acquisition efficiency. Together, the graphic and tabular views in the ABP Visualizer provide a flexible and effective interface for pathway relation browsing and analysis. Our study contributes to pathway-related research and biological information extraction by assessing the value of a multiview, relation-based interface that supports user-controlled exploration of pathway information across multiple granularities. PMID- 17912970 TI - Variable background active contour model for computer-aided delineation of nodules in thyroid ultrasound images. AB - This paper presents a computer-aided approach for nodule delineation in thyroid ultrasound (US) images. The developed algorithm is based on a novel active contour model, named variable background active contour (VBAC), and incorporates the advantages of the level set region-based active contour without edges (ACWE) model, offering noise robustness and the ability to delineate multiple nodules. Unlike the classic active contour models that are sensitive in the presence of intensity inhomogeneities, the proposed VBAC model considers information of variable background regions. VBAC has been evaluated on synthetic images, as well as on real thyroid US images. From the quantification of the results, two major impacts have been derived: 1) higher average accuracy in the delineation of hypoechoic thyroid nodules, which exceeds 91%; and 2) faster convergence when compared with the ACWE model. PMID- 17912971 TI - Strong compound-risk factors: efficient discovery through emerging patterns and contrast sets. AB - Odds ratio (OR), relative risk (RR) (risk ratio), and absolute risk reduction (ARR) (risk difference) are biostatistics measurements that are widely used for identifying significant risk factors in dichotomous groups of subjects. In the past, they have often been used to assess simple risk factors. In this paper, we introduce the concept of compound-risk factors to broaden the applicability of these statistical tests for assessing factor interplays. We observe that compound risk factors with a high risk ratio or a big risk difference have an one-to-one correspondence to strong emerging patterns or strong contrast sets-two types of patterns that have been extensively studied in the data mining field. Such a relationship has been unknown to researchers in the past, and efficient algorithms for discovering strong compound-risk factors have been lacking. In this paper, we propose a theoretical framework and a new algorithm that unify the discovery of compound-risk factors that have a strong OR, risk ratio, or a risk difference. Our method guarantees that all patterns meeting a certain test threshold can be efficiently discovered. Our contribution thus represents the first of its kind in linking the risk ratios and ORs to pattern mining algorithms, making it possible to find compound-risk factors in large-scale data sets. In addition, we show that using compound-risk factors can improve classification accuracy in probabilistic learning algorithms on several disease data sets, because these compound-risk factors capture the interdependency between important data attributes. PMID- 17912972 TI - Incremental diagnosis method for intelligent wearable sensor systems. AB - This paper presents an incremental diagnosis method (IDM) to detect a medical condition with the minimum wearable sensor usage by dynamically adjusting the sensor set based on the patient's state in his/her natural environment. The IDM, comprised of a naive Bayes classifier generated by supervised training with Gaussian clustering, is developed to classify patient motion in-context (due to a medical condition) and in real-time using a wearable sensor system. The IDM also incorporates a utility function, which is a simple form of expert knowledge and user preferences in sensor selection. Upon initial in-context detection, the utility function decides which sensor is to be activated next. High-resolution in context detection with minimum sensor usage is possible because the necessary sensor can be activated or requested at the appropriate time. As a case study, the IDM is demonstrated in detecting different severity levels of a limp with minimum usage of high diagnostic resolution sensors. PMID- 17912973 TI - Analysis of dynamic susceptibility contrast MRI time series based on unsupervised clustering methods. AB - We compare five different unsupervised clustering techniques as tools for the analysis of dynamic susceptibility contrast MRI time series. The study included four subjects: two subjects with stroke and two subjects without focal neurological deficit. The goal was to determine the robustness and reliability of clustering methods in providing a self-organized segmentation of perfusion MRI data sharing common properties of signal dynamics. For this purpose, the relative signal reduction time series was computed for each pixel. Clustering of the resulting high-dimensional feature vectors was performed by minimal free-energy deterministic annealing, self-organizing maps, two variants of fuzzy c-means clustering (FVQ and FSM), and the neural gas algorithm. Clustering results were evaluated by visual assessment of cluster assignment maps and corresponding signal time curves as well as by quantitative comparison of cluster assignment maps with conventional pixel-specific perfusion parameter maps based on quantitative receiver operating characteristic (ROC) curve analysis. Clustering methods provided a functional segmentation with respect to vessel size, detected side asymmetries of contrast-agent first pass, and identified regions of perfusion deficits in subjects with stroke. As confirmed by quantitative ROC analysis, the clustering approach can detect regions of reduced brain perfusion with high accuracy when compared to conventional analysis by pixel-specific cerebral blood volume and mean transit time maps. We conclude that by unveiling differences of signal dynamics and amplitude, clustering is a useful tool to analyze and visualize regional properties of brain perfusion. Thus, it may contribute to the computer-aided diagnosis of cerebral circulation deficits by noninvasive neuroimaging. PMID- 17912974 TI - ELB-Q: a new method for improving the robustness in DNA microarray image quantification. AB - Reliable and robust quantification of signal intensities is a critical step in microarray-based biomedical studies. However, traditional techniques for microarray image processing would face significant challenges if the number of pixels used for the quantification of the local background and the foreground decreases dramatically. We have developed a new method, ELB-Q, which, by design, is well suited for the image quantification of microarrays with very high density of spot layout (large number of spots arranged in unit area). In ELB-Q, a large extended local background (ELB) interspot region excluding those "noise of the background" pixels is used for estimating the local background, and the quantification of spot intensities (mean and median) in the putative target spot regions is performed after further excluding background pixels in these areas based on the cutoff values established during the ELB calculation. ELB-Q takes advantage of the abundant spatial information around each spot of interest, makes no assumption of the shape and size of the spots, and needs no sophisticated adjustment. We show results of image processing using ELB-Q on both the simulated data and real DNA microarrays, which compare favorably in robustness and accuracy against those obtained with GenePix Pro 6.0 (Axon Instruments, 1999) and the Markov random field (MRF) modeling approach. The ELB-Q software is developed in Matlab, and is available upon request. PMID- 17912975 TI - A contextual medical image viewer. AB - The conventional visualization of medical images is not enough for a rich and comprehensive electronic healthcare record. We believe that it is necessary to provide a viewer with more advanced capabilities than those of regular medical image viewers. In this paper, we propose an architecture that allows the use of contextual information to assist the healthcare professional in his regular tasks. The proposed architecture for the context is composed of an ontology describing the hospital and an inference engine. The result is a Java-based implementation of the architecture, named Dynamic N-dimensional Image Viewer, (ODIN in Portuguese) that is able to adapt its behavior according to the contextual information. PMID- 17912976 TI - A probabilistic peptide machine for predicting hepatitis C virus protease cleavage sites. AB - Although various machine learning approaches have been used for predicting protease cleavage sites, constructing a probabilistic model for these tasks is still challenging. This paper proposes a novel algorithm termed as a probabilistic peptide machine where estimating probability density functions and constructing a classifier for predicting protease cleavage sites are combined into one process. The simulation based on experimentally determined Hepatitis C virus (HCV) protease cleavage data has demonstrated the success of this new algorithm. PMID- 17912977 TI - Visualization of back pain data--a 3-D solution. AB - Traditional approaches to gathering and visualizing pain data rely on two dimensional (2-D) human body models, where different types of sensation are recorded with various monochrome symbols. We propose an alternative that uses a three-dimensional (3-D) representation of the human body, which can be marked in color to visualize and record pain data. PMID- 17912978 TI - Collaborative work during interventional radiological procedures based on a multicast satellite-terrestrial network. AB - Collaboration is a key requirement in several contemporary interventional radiology procedures (IRPs). This work proposes a multicast hybrid satellite system capable of supporting advanced IRP collaboration, and evaluates its feasibility and applicability. Following a detailed IRP requirements study, we have developed a system which supports IRP collaboration through the employment of a hybrid satellite-terrestrial network, a prototype multicast version of wavelet based interactive communication system (WinVicos) application, and a partition aggregation and conditional coding (PACC) wavelet codec. A semistructured questionnaire was also used to receive evaluative feedback from collaborating participants. The departments of interventional radiology of University Hospital of Patras, Greece and of Charite Hospital of Berlin, Germany have been connected on the system. Eight interventional radiologists and a vascular surgeon participated periodically in three satellite-terrestrial "fully collaborative" IRPs (average time 90 min) of high complexity and in four terrestrial educational sessions with great success, evidenced by considerable improving the IRP outcomes (clinical and educational). In case of high complexity, where the simultaneous presence of remote interventional expert and/or surgeon is required, advanced collaboration among staff of geographically dispersed international centers is feasible via integration of existing networking and other technologies. PMID- 17912979 TI - Acute hepatitis A in children. PMID- 17912980 TI - Umbilical cord blood transplantation: application in pediatric patients. AB - Although allogeneic stem cell transplantation can cure patients with hematologic malignancies and nonmalignant disorders, limiting factors such as lack of suitable donors and graft-versus-host disease (GVHD) toxicity have led to the exploration of umbilical cord blood (UCB) as an alternative source of hematopoietic stem cells. The unique immunologic properties of UCB likely contribute to a decreased risk of GVHD. Thus, UCB represents a highly convenient hematopoietic stem cells (HSC) source that may significantly expand the HSC donor pool. This overview examines the advantages and disadvantages of UCB as a donor source and outlines the current status of UCB transplantation (UCBT), with emphasis on pediatric practice. Strategies to improve kinetics of hematopoietic recovery after UCB grafting in children and adults are warranted. PMID- 17912981 TI - Childhood occipital epileptic syndromes in Taiwan: clinical characteristics and outcomes. AB - BACKGROUND: The present study aimed to elucidate the clinical characteristics and outcomes of three types of occipital epileptic syndromes: early-onset and late onset childhood epilepsy with occipital paroxysm (ECEOP and LCEOP) and symptomatic occipital epilepsy (SOLE). METHODS: We retrospectively reviewed the medical records of 54 children (18 ECEOP, 10 LCEOP, and 26 SOLE) and compared the clinical features, EEG findings, treatments, and outcomes among these patients. RESULTS: Nocturnal seizures occurred in 55% of the patients with ECEOP, whereas of those with LCEOP and SOLE, 80% and 61% had diurnal seizures, respectively (P = 0.04). Status epilepticus was more common in the ECEOP group, less in LCEOP. Autonomic auras were more common in the SOLE group than in the other two groups. Secondary generalized seizure was often found in the SOLE group (P = 0.03). EEG findings in 72% of the ECEOP group, 60% of the LCEOP group, and 19.2% in the SOLE group were changeable and became to be normal after 5 years of follow-up. Slow EEG background activity was more evident in the SOLE group than in the other two groups. The prognoses of these groups of patients were different. The ECEOP group had the best, and the SOLE group had the worst. CONCLUSIONS: These findings suggest that some clinical features of these three syndromes differ from each other, which may provide clinicians a basis for determining the appropriate diagnosis in children with one of these childhood occipital epileptic syndromes. PMID- 17912982 TI - Clinical and laboratory characteristics of type 1 diabetes in children and adolescents: experience from a medical center. AB - BACKGROUND: The incidence of type 1 diabetes (T1D) is increasing rapidly worldwide, predominantly in younger individuals. We developed a checklist of all symptoms of T1D reported in the literature and compared the completeness of the recording of symptoms at initial presentation before and after the checklist was adopted. METHODS: We retrospectively reviewed the records of patients newly diagnosed with T1D from January 1, 1979 through September 30, 2006 to assess the presenting features and test the usefulness of a symptom checklist in evaluating the history on presentation. The checklist was incorporated into the records as of October 1, 1994. RESULTS: Of the 304 patients identified, 130 (43%) had checklists in the charts. There were 146 (48%) boys, 98 (32%) who were diagnosed under the age of 6 years, and 198 (65%) presented with diabetic ketoacidosis (DKA). Records with a checklist noted diabetic symptoms that were subtle and easily ignored more often than records without the checklist. As compared with those diagnosed at an older age, patients diagnosed at < or = 6 years were more likely to be male, have DKA and a shorter symptom duration, and report more episodes of preceding viral infection and dyspnea. Patients with DKA also had a shorter symptom duration. CONCLUSIONS: A diabetic symptom checklist was helpful in identifying clinical diabetic symptoms and signs which were otherwise easily ignored. Younger children were more likely to have a shorter symptom duration and a higher incidence of DKA. PMID- 17912983 TI - Discriminant analysis of serum inflammatory biomarkers which differentiate pediatric appendicitis from other acute abdominal diseases. AB - BACKGROUND: Other acute abdominal diseases in children can confound the signs and symptoms of appendicitis, resulting in misdiagnosis and unnecessary appendectomy. In this study, we used discriminant analysis of serum inflammatory biomarkers to determine which acute abdominal diseases could mimic appendicitis, and further analyzed these diseases based on different age groups. METHODS: We prospectively collected 417 patients aged from 4 to 18 years with clinically suspected acute appendicitis in the pediatric emergency department. We selected the significantly higher serum biomarkers for appendicitis as the discriminating variables. Furthermore, we analyzed the definitive diagnoses of patients with normal appendices who could not be predicted by discriminant analysis. RESULTS: Patients with acute appendicitis had significantly higher leukocyte counts (p < 0.01), neutrophil counts, (p < 0.01) and C-reactive protein concentrations (p < 0.01 ) than those with normal appendices. The discriminant power of these three serum biomarkers in acute appendicitis was 76 percent. Acute abdominal diseases which mimicked appendicitis included acute gastroenteritis, nonspecific abdominal pain, urinary tract infection, and upper respiratory infection with gastrointestinal upset. CONCLUSIONS: Serum biomarkers may serve as helpful discriminators to predict the presence of pediatric appendicitis. But, some acute abdominal diseases mimicking appendicitis should be considered during differential diagnosis of acute appendicitis to avoid making misdiagnosis and performing unnecessary appendectomy. PMID- 17912984 TI - Acute hepatitis A infection in children: a 20-year experience of a medical center in Southern Taiwan. AB - BACKGROUND: Taiwan is currently nonendemic for hepatitis A infection, but sporadic cases are reported occasionally. The presenting symptoms of acute hepatitis A (AHA) infection in children are often nonspecific and may be easily misdiagnosed as general gastrointestinal illness. METHODS: We conducted this study to identify the relevant presenting symptoms and laboratory findings of AHA in children. The medical records of all patients younger than 18 years old admitted to Kaohsiung Chang Gang Memorial Hospital from July 1986 to June 2004 under the diagnosis of AHA were reviewed. There were 29 pediatric patients admitted with the diagnosis of AHA, including 16 male patients and 13 female patients. RESULTS: Poor appetite, malaise, abdominal pain, vomiting and fever were the leading constitutional symptoms. Jaundice and tea-colored urine were the most relevant hepatobiliary presentations. Elevation of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, direct and indirect bilirubin was common, although other hematology and biochemistry examinations were within normal limits on most occasions. There was no mortality in our series. A slightly increasing prevalence of AHA was noted over the two decades of the study period, although it was not statistically significant. CONCLUSIONS: Our results found that children with AHA tended to present with non specific gastrointestinal symptoms and acute liver function deterioration. Elevation of hepatic aminotransferase and alkaline phosphatase with cholestasis was common. PMID- 17912985 TI - Perinatal and infant health outcomes among neonates born to aboriginal parents in Taiwan. AB - BACKGROUND: Poor health outcomes among neonates born to aborigines has been reported in many countries. This study was aimed to examine the nationwide characteristics of live births, adverse birth outcomes, and age-specific mortality among neonates born to non-aboriginal and aboriginal parents in Taiwan. METHODS: All neonates born alive during the period of 2000 to 2003 in Taiwan were included. The adverse birth outcomes including low birth weight, preterm, and small-for-gestational-age births, and age-specific mortality were obtained. Logistic regression analysis was used to estimate odds ratios for parental ethnicity in relation to birth outcomes, while Cox's proportional hazards regression models were used to estimate hazard ratios for parental ethnicity in relation to age-specific infant deaths. RESULTS: A total of 947,317 live births were included that consisted of 9,381 born to aboriginal mothers, 6,429 born to aboriginal fathers, and 15,354 born to aboriginal parents. There was a gradual increase in the risk of having a baby with low birth weight, preterm, or small for gestational age born to the four parental aboriginal ethnicity groups: non aboriginal parents, aboriginal mother only, aboriginal father only, and aboriginal parents. Similar trends were also found for early neonatal, neonatal, and infant mortalities after stratification of residential areas. The neonates born to both aboriginal parents with residence in rural or mountain areas were at highest risk of adverse birth outcomes and age-specific mortality. CONCLUSIONS: Our results demonstrated that aboriginality and residential area are important risk factors for adverse perinatal and infant outcomes. PMID- 17912986 TI - Outcome of myasthenia gravis mothers and their infants. AB - BACKGROUND: The aim of this retrospective study was to assess neonatal outcomes of pregnant women with myasthenia gravis (MG). METHODS: Pregnant women with MG who were treated in our hospital over an 8-year period were enrolled. Data relating to the course of the mother's MG (including the anti-acetylcholine receptor antibody (anti-AchR) titer and drug dosage), delivery mode, delivery course, puerperium period, and neonatal outcomes were obtained from the medical records. RESULTS: Twelve women with MG had 13 pregnancies in our hospital from January 1997 to December 2005. None of the mothers needed intensive care. Two patients delivered vaginally, and ten delivered by cesarean section. Fourteen infants were born at an average gestational age of 37.2 +/- 2.0 weeks; their average birth weight was 2838.6 +/- 724.2 g. Two neonates (14.2%) had a congenital anomaly. Transitory neonatal myasthenia gravis (NMG) was diagnosed in one infant (7.1%). CONCLUSIONS: In our study, MG exacerbations occurred in 38.5% of the patients. Postpartum, all MG patients experienced deterioration except one patient who deteriorated in the first trimester with a concomitant upper airway tract infection. Only 1 of the 14 neonates developed transient NMG; the incidence of transient NMG was lower than that previously reported. There was no correlation between the occurrence of NMG and the maternal anti-AChR titer. The cesarean section rate was approximately 33%; this high rate of elective cesarean sections in these MG patients could have prevented the occurrence of some of the complications related to vaginal delivery. PMID- 17912987 TI - Steroid use in sepsis-induced conjugated hyperbilirubinemia: report of one case. AB - We report a girl with acute lymphoblastic leukemia who developed severe conjugated hyperbilirubinemia (total bilirubin 34.32 mg/dl, direct bilirubin 26.94 mg/dl) following gram-negative (Escherichia coli ) sepsis. Despite broad spectrum antibiotics, her hyperbilirubinemia progressed. Her mental status deteriorated and steroid therapy (hydrocortisone 8 mg/kg/d) was initiated. Her condition then recovered rapidly. Based on our findings, we suggest that steroid may help to treat progressive conjugated hyperbilirubinemia due to gram-negative sepsis. PMID- 17912988 TI - Recurrent Kimura's disease: report of one case. AB - Kimura's disease (KD) is a rare, chronic inflammatory disease, and the etiology is not yet to be determined. The disease is usually found in young Asian men but rarely in Western population. Here, we report an 8-year-old boy with the initial presentation of right upper eyelid swelling. Laboratory tests disclosed peripheral eosinophilia and elevated IgE level. KD with lacrimal gland involvement was diagnosed. Surgical excision was performed several times and steroid was also used, however the disease recurred. Thereafter, steroid, cyclosporine-A and azathioprine were prescribed. The tumor mass resolved and the laboratory data returned to normal level. PMID- 17912989 TI - Accelerated baccalaureate programs: what we know and what we need to know- setting a research agenda. PMID- 17912990 TI - Second-career baccalaureate nursing students: a lived experience. AB - Second-career baccalaureate nursing programs and the students enrolled in them have been a topic of interest since these programs were first introduced into the academic setting in the 1970s. The purpose of this phenomenological inquiry was to develop an understanding of the meaning of the lived experience of being a second-career baccalaureate nursing student. Five second-career baccalaureate nursing students participated in unstructured, in-depth, face-to-face interviews. Data analysis was guided by the phenomenological method of van Manen. The themes identified were Questioning One's Place in the World; Seeing One's Place in the World in Another Way; Preparing for the Plunge; Trying Transitions; A Bundle of Emotions; Faculty Control, Student Imbalance; and Almost There and Scared. Implications of this research related to curriculum revision are presented. PMID- 17912991 TI - Accelerated second-degree baccalaureate nursing programs: what is the significance of clinical instructors? AB - Accelerated second-degree nursing programs are a popular method of entry into nursing for many individuals beginning a second career in nursing. Using a hermeneutic phenomenological approach, this study analyzed the experiences of 19 accelerated second-degree baccalaureate nursing graduates to understand how their student experiences best prepared them for nursing practice after graduation. The significance of clinical instructors to students' development as nurses was revealed in the data analysis and identified in the theme, Blending Practice and Pedagogy. This article describes the implications of this theme for both accelerated second-degree nursing programs and faculty recruitment and retention. PMID- 17912992 TI - Use of self-regulating learning strategies by students in the second and third trimesters of an accelerated second-degree baccalaureate nursing program. AB - Accelerated nursing programs have proliferated in recent years, attracting adult learners with varied backgrounds and experience into the profession of nursing. Learner adaptation to and successful completion of nursing programs delivered in an accelerated format require that students use self-regulatory learning strategies to focus, organize, integrate, and retain knowledge derived from classroom and clinical sources. The purpose of this descriptive, exploratory study was to examine the presence and extent of and differences in self-reported regulatory learning strategy use in a convenience sample of two groups of non nursing college graduates. Determination of strategy use was delineated by student responses to the learning strategy subscales of the Motivated Strategies for Learning Questionnaire. Results of this study demonstrate that although both groups of students used self-regulated learning strategies, students who have completed more of the accelerated program use more self-regulatory learning strategies than their less experienced counterparts. PMID- 17912993 TI - The value of admission interviews in selecting accelerated second-degree baccalaureate nursing students. AB - Accelerated undergraduate nursing programs have attracted more qualified applicants than can be accommodated, yet higher-than-desired attrition rates continue for a variety of reasons (e.g., pace of program, poor lifestyle choices, role concerns). The financial effects of the loss of students from these programs has gone largely undocumented and is intensified by the knowledge that a better admission decision could have resulted in another nurse in the workforce. In this article, the inclusion of structured personal interviews into the admission decision-making process is discussed. Specifically, do interviews provide relevant data in making admission decisions? Attempts to reduce subjectivity through the development of a structured interview guide, the training of paired interview teams, and the institution of a "post-mortem" technique to guide revisions are also described. Interviews do result in denial of applicants who seem appropriate based on paper portfolio criteria, allowing for inclusion of other potentially successful applicants. PMID- 17912995 TI - Avoiding socialization pitfalls in accelerated second-degree nursing education: the returning-to-school syndrome model. AB - Second-degree students are highly motivated and tend to excel academically. However, nurse educators in accelerated programs face challenges in socializing these students to the nursing role. One pitfall is the hostility that may develop if students perceive a mismatch between their expectations and their new role as baccalaureate nursing (BSN) students in a fast-paced and intense program. This article discusses the applicability of the returning-to-school syndrome model in helping second-degree nursing students maneuver successfully through an accelerated BSN program. This 3-stage model has been previously applied in RN-to BSN education. Using the model in an accelerated BSN curriculum to identify transition points and offer student support through specific stages can better prepare students to meet the challenges of accelerated education, as well as help nurse educators become more adept at providing resources and implementing supportive strategies at the appropriate time. PMID- 17912994 TI - Designing hybrid web-based courses for accelerated nursing students. AB - Due to a heightened interest in accelerated nursing programs, enrollment in schools of nursing is rapidly increasing. Nursing programs need to find more creative ways to reach and teach older, technologically savvy adult learners, who have different learning needs than traditional undergraduate nursing students. These students present new challenges to nursing faculty. Larger class sizes, prior academic accomplishments, and a shorter time frame for assimilation of the nursing role provide opportunities for creative course delivery. This article discusses how a change in course design, specifically a hybrid Web-based course, can address these challenges. PMID- 17912996 TI - Service-learning across an accelerated curriculum. AB - In support of the community-based education philosophy, nursing faculty at a small health careers college in the midwestern United States incorporated a service-learning experience involving Sudanese refugee families and Latino community members into its accelerated community-based nursing program (ACE) curriculum. The purpose of the service-learning experience is to allow students the opportunity develop relationships through which they may gain an understanding of unique needs within a community. This article describes the development and benefits of the service-learning experience across the ACE curriculum. Through reflection, students broaden and deepen their understanding about the health of individuals in the community and acquire a more sophisticated understanding of risk PMID- 17912997 TI - Comparative analysis between hard- and soft-filament toothbrushes related to plaque removal and gingival abrasion. AB - OBJECTIVE: The aim of this split-mouth, examiner-blind study was to compare the dental plaque removal and incidence of gingival abrasion associated with the use of hard- and soft-filament toothbrushes. METHODOLOGY: The test group consisted of 20 non-dental students, mean age 25 years. After a three-day period of plaque accumulation following the use of a disclosing solution, the Quigley-Hein Plaque Index was recorded, while the presence of gingival abrasion was measured from photographs. Pairs of quadrants 1-3 and 2-4 were allocated to supervised brushing with hard- or soft-filament toothbrushes for 30 seconds, limited to the buccal aspects of the teeth. Plaque levels and gingival abrasion were again assessed. Initial and final values of the plaque index and the mean number of abrasions were compared with the Friedman and Wilcoxon tests (p < or = 0.05). RESULTS: Plaque indices were reduced significantly from a baseline of 4.12 in both groups to 1.21 after the use of hard-filament toothbrushes, and to 1.67 after the use of soft-filament toothbrushes. The use of hard-filament toothbrushes resulted in a significantly higher mean number of lesions when compared to the soft-filament toothbrushes; 11.6 and 7.9, respectively (p = 0.018). CONCLUSION: Hard-filament toothbrushes remove more plaque than soft filament brushes, but also cause a higher number of gingival abrasions. PMID- 17912998 TI - Efficacy and safety of a new power toothbrush in a population with mild to moderate gingivitis. AB - OBJECTIVE: To evaluate the efficacy and safety of a novel ultrasound power toothbrush over a 30-day period in a population with mild to moderate gingivitis. METHODOLOGY: A 30-day, randomized, examiner-blind, parallel-arm study was conducted to evaluate the safety and efficacy of a ultrasound novel power toothbrush (Ultreo - ULT) as compared to a control manual toothbrush (Oral-B 35 MTB). Fifty-three subjects (n = 26 ULT, n = 27 MTB) with a minimum of 18 natural teeth and a Loe and Silness Gingival Index of > or = 1.5 were enrolled. An intraoral examination (oral soft and hard tissues, restorations) was performed, and Loe and Silness Gingival Index scores were recorded at baseline and 30 days. Subjects were instructed to brush at home twice per day with their assigned toothbrush and study toothpaste. A product evaluation questionnaire was also completed at the 30-day study visit. RESULTS: Oral examinations indicated normal findings at all time points for both groups, and no adverse events were reported during the study. There were no significant differences in gingivitis scores at baseline between the toothbrush groups (p > 0.05). From baseline, each treatment group demonstrated a significant reduction in gingivitis over the 30-day period (p < 0.001). However, subjects in the ULT group demonstrated a significantly greater reduction in gingivitis compared to those using the manual toothbrush (p = 0.010). Results from the questionnaire, on average, indicated subjects in the ULT group scored their device higher with respect to the clean feeling after brushing and perceived improvements in gingival health. CONCLUSION: The novel Ultreo ultrasound power toothbrush significantly reduced gingivitis over the 30 day study, and resulted in a significantly greater reduction in gingivitis than a manual toothbrush. No adverse events or safety concerns were identified with either the power or manual toothbrush. PMID- 17912999 TI - Evaluation of the antibacterial activity of new root canal sealers. AB - OBJECTIVE: The purpose of this study was to compare the antimicrobial effect of AH-26, RealSeal (RS), and GuttaFlow after 24 hours and after one week. METHODOLOGY: All sealers were prepared according to the manufacturers' directions immediately before testing. The microorganisms used in this assay included Staphylococcus aureus and Streptococcus mutans, which were prepared from isolated species in the microbiology laboratory. The antimicrobial effects of each sealer were determined by measuring the diameter of the zone of inhibition in millimeters after incubation at 37 degrees C for 24 hours and for seven days in a humid atmosphere. Each test was repeated three times. RESULTS: In the 24-hour samples, as well as in seven-day samples, the antibacterial activity of AH-26 was significantly greater than two other materials tested against both bacteria (p < 0.05). The antibacterial activities of RS and GuttaFlow against Staphylococcus aureus were not significantly different from each other (p > 0.05) in 24-hour as well as seven-day samples, but RS had significantly greater effect against Streptococcus mutans than GuttaFlow (p < 0.05). CONCLUSION: The products studied exhibited some variable antibacterial properties. In 24 hours, as well as in seven days, the AH-26 exerted the greatest activity against both organisms tested. PMID- 17913000 TI - Manual toothbrush wear and consequences on plaque removal. AB - OBJECTIVE: The aim of this study was to determine the mean time interval before needing to discard different types of manual toothbrushes based on wear, and on the impact of progressive wear on plaque removal efficacy. METHODOLOGY: Two cross over randomized clinical trials involving the same 12 volunteers tested four toothbrushes of different designs. In Study 1, the brushes were the Butler 211 (BTB) and Fluocaril Sensia (FSTB). In Study 2, the brushes were Elmex InterX (ETB) and Elgydium Interactive (ELTB). In both trials, the volunteers randomly used one of the two toothbrushes for two minutes twice a day during the first week, and the other brush during the following week, and continued this alternating routine for two months. An image acquisition system and the Visilog 5.2 image analysis program were used to calculate the wear index (WI) after one and two months of use for each toothbrush. The OHI-S plaque index (PI) was recorded after one and three minutes of tooth brushing at one and two months. The Friedman or Kruskal-Wallis test was used to compare data. RESULTS: Except for the ELTB, the wear of the toothbrushes was greater during the first month of brushing than during the second. The total WI was FSTB 35.46 (range = 62.11), ELTB 30.14 (range = 67.92), ETB 48.77 (range = 123.87), and BTB 98.20 (range = 134.75). The differences were significant at p = 0.008. The post one-minute PI scores significantly increased with wear, except in the case of ELTB (p = 0.244). When comparing all toothbrushes' PI at the one and three-minute tooth brushing times, there were significant differences at one month among them at p < 0.0001. CONCLUSION: The amount of wear depended on the toothbrush design. It was greater for rectangular tooth brushing areas or when bristles had smaller diameters and different heights. With two-month-old toothbrushes, plaque removal was better when there was less measurable wear. PMID- 17913001 TI - Anticalculus effect of a cetylpyridinium chloride/zinc gluconate mucoadhesive gel: results of a randomized, double-blind, controlled clinical trial. AB - OBJECTIVE: This randomized, double-blind, controlled clinical trial investigated the anticalculus effect of a cetylpyridinium chloride/ zinc gluconate (CPC/ZG) mucoadhesive gel. METHODOLOGY: The 80 adults from the San Francisco area who fulfilled the enrollment criteria were stratified based on total Volpe-Manhold Index (VMI) scores (low, medium, or high), gender, and other demographic data. Within these strata, they were randomly assigned to the CPC/ZG gel group or the placebo gel group, and underwent a baseline oral soft tissue (OST) examination. Subjects were dispensed Oral-B Indicator 35 Compact Head soft toothbrushes and Colgate Cavity Protection MFP toothpaste, and instructed to brush twice daily using these products. Every night for three months, following the nighttime brushing and just before retiring, subjects applied their assigned treatment gel to the lingual surfaces of the six mandibular anterior teeth. They were instructed not to eat or drink until morning and to avoid using other methods of interdental cleaning between the treated teeth, except to remove impacted food. Total VMI scores and OST examination findings obtained at three months were compared with baseline findings. RESULTS: Seventy-eight (78) of the 80 subjects complied with the protocol and completed the study. After three months of treatment, the CPC/ZG group showed a 30% decrease in mean VMI score compared with a 0% decrease in the placebo group. OST examination at three months revealed no serious adverse events in either group. CONCLUSION: Results of this clinical trial indicate that regular use of the CPC/ZG gel, with or without the use of floss or other interdental cleaning products, yields a statistically significant reduction in calculus. PMID- 17913003 TI - Effects of hydrogen peroxide bleaching strips on root surfaces in vitro. AB - OBJECTIVE: This study examined the effects of hydrogen peroxide tooth whiteners on the physical hardness and morphology of human root dentin cycled through bleaching treatments in vitro. METHODOLOGY: Human root dentin was ground and polished to prepare a uniform substrate for bleaching treatments. Half the root dentin specimens were acid (EDTA) etched to remove the smear layer, while the other half were treated with an intact smear layer. A cycling treatment methodology was employed which alternated ex vivo human salivary exposures with bleaching treatments under conditions of controlled temperature and durations of treatment. Bleaching treatments included Crest Whitestrips bleaching gels, which utilize hydrogen peroxide as the in situ bleaching source at 5.3 and 6.5% concentrations of hydrogen peroxide, placebo gels containing no hydrogen peroxide, and an untreated control group. Surface color measurements were taken prior to and following bleaching. Effects of bleach on physical properties of dentin were assessed with microhardness, and surface morphology effects were studied with scanning electron microscopy (SEM). RESULTS: Color assessments confirmed significant ex vivo tooth bleaching by treatments. Surface microhardness values remained the same or increased slightly with bleaching treatments for both smear layer and etched dentin surfaces. SEM measures of non etched dentin surfaces cycled with bleach showed preservation of the dentin smear layer. SEM of etched dentin treated with bleach showed maintenance of standard tubule diameters. CONCLUSION: These results confirm that tooth bleaching with hydrogen peroxide bleaching gels at 5.3 and 6.5% concentrations do not produce solubilization reactions on dentin surfaces in vitro, and that dentin smear layers remain intact following bleaching exposures. PMID- 17913002 TI - A new mouthrinse combining zinc and chlorhexidine in low concentrations provides superior efficacy against halitosis compared to existing formulations: a double blind clinical study. AB - OBJECTIVE: Volatile sulfur compounds (VSC), mainly derived from bacteria located in deep crypts at the back of the tongue and from periodontal pockets, are responsible for approximately 90% of halitosis (bad breath, malodor). The objective of this double blind clinical study was to assess the clinical efficacy of a new formulation for halitosis containing a combination of zinc (0.3% Zn) and chlorhexidine (0.025% CHX) in low concentrations. The new formulation was compared to some widely used and commercially available formulations containing various enzymes and antibacterial agents in a clinical setting under controlled conditions. METHODOLOGY: Ten healthy volunteers participated in this study (5 female, 5 male, mean age: 46.6, range: 26-79). Each participant served as their own control, and neither the investigator nor the ten test subjects knew which formulation they were testing at any given time (double-blind design). Baseline H2S data were obtained by cysteine rinsing for 30 seconds, 90 seconds mouth closure, and gas chromatographic (GC) analysis of mouth air. On separate days, each participant then rinsed for 60 seconds with 10 ml of each of the eight various formulations. Cysteine rinses were repeated at 1 hour, 2 hours, and 3 hours, and GC measurements of oral H2S levels were again recorded. RESULTS: The test rinse (0.3% Zn + 0.025% CHX) reduced the intraoral H2S levels to 0.16% of control (range: 0.01-0.54%) after 1 hour, 0.4% after 2 hours, and 0.75% after 3 hours, providing superior efficacy in reducing H2S compared to the other formulations tested (p < 0.05). CONCLUSION: A combination of Zn and CHX in low concentrations seems to be the most efficient way to remove the VSC that causes bad breath at present. Studies are underway to further explore the extraordinary efficacy of this combination (close to 100%), suggesting a specific mode of action and a synergistic effect of these two components. PMID- 17913004 TI - Development of the Evolve toothbrush: laboratory evaluations of gingival margin cleaning and subgingival access. AB - OBJECTIVE: The Evolve toothbrush has been developed with three heads that adjust to the tooth shape with brushing. With the exertion of gentle pressure on the center head against the tooth cusps, the two outer heads bend and conform to the tooth anatomy at the gingival areas around each tooth. In preliminary clinical observations, decreased plaque and improved gingival appearance was observed after one and two weeks of unsupervised product use. The purpose of this laboratory research was to evaluate the ability of the Evolve toothbrush bristles to remove artificial plaque deposits at and around the gingival margin (Gingival Margin Cleaning, GMC), and to gain access into and remove artificial plaque deposits from under the gingival margin (Subgingival Access, SA) using standardized, published methods. METHODOLOGY: In the first two laboratory studies conducted, two toothbrush products, the new Evolve and the standard American Dental Association toothbrush (ADA) were evaluated for GMC and SA. In the third assay, three toothbrushes, the Evolve, Oral-B CrossAction, and the Sonicare Elite, were evaluated for SA. Each toothbrush product group was evaluated for 24 assessments using clinical tooth brushing motions on posterior tooth shapes under wet brushing conditions. RESULTS: In all laboratory tests conducted, the Evolve toothbrush had significantly (p < 0.001) greater efficacy compared to the other toothbrushes tested. CONCLUSION: The Evolve toothbrush warrants further clinical evaluations as it is believed these future findings will further substantiate those of the initial testing, and demonstrate the Evolve's ability to help improve oral and overall health. PMID- 17913005 TI - Paradox found (again): infant mortality among the Mexican-origin population in the United States. AB - Recent research suggests that the favorable mortality outcomes for the Mexican immigrant population in the United States may largely be attributable to selective out-migration among Mexican immigrants, resulting in artificially low recorded death rates for the Mexican-origin population. In this paper we calculate detailed age-specific infant mortality rates by maternal race/ethnicity and nativity for two important reasons: (1) it is extremely unlikely that women of Mexican origin would migrate to Mexico with newborn babies, especially if the infants were only afew hours or afew days old; and (2) more than 50% of all infant deaths in the United States occur during the first week of life, when the chances of out-migration are very small. We use concatenated data from the U.S. linked birth and infant death cohort files from 1995 to 2000, which provides us with over 20 million births and more than 150,000 infant deaths to analyze. Our results clearly show that first-hour, first-day, and first-week mortality rates among infants born in the United States to Mexican immigrant women are about 10% lower than those experienced by infants of non-Hispanic, white U.S.-born women. It is extremely unlikely that such favorable rates are artificially caused by the out-migration of Mexican-origin women and infants, as we demonstrate with a simulation exercise. Further, infants born to U.S.-born Mexican American women exhibit rates of mortality that are statistically equal to those of non-Hispanic white women during the first weeks of life and fare considerably better than infants born to non-Hispanic black women, with whom they share similar socioeconomic profiles. These patterns are all consistent with the definition of the epidemiologic paradox as originally proposed by Markides and Coreil (1986). PMID- 17913006 TI - Chronic conditions and the decline in late-life disability. AB - Using data from the 1997-2004 National Health Interview Survey (NHIS), we examine the role of chronic conditions in recent declines in late-life disability prevalence. Building upon prior studies, we decompose disability declines into changes in the prevalence of chronic conditions and in the risk of disability given a condition. In doing so, we extend Kitigawa's (1955) classical decomposition technique to take advantage of the annual data points in the NHIS. Then we use respondents' reports of conditions causing their disability to repartition these traditional decomposition components. We find a general pattern of increasing prevalence of chronic conditions accompanied by declines in the percentage reporting disability among those with a given condition. We also find declines in heart and circulatory conditions, vision impairments, and possibly arthritis and increases in obesity as reported causes of disability. Based on decomposition analyses, we conclude that heart and circulatory conditions as well as vision limitations played a major role in recent declines in late-life disability prevalence and that arthritis may also be a contributing factor. We discuss these findings in light of improvements in treatments and changes in the environments of older adults. PMID- 17913007 TI - Was there compression of disability for older Americans from 1992 to 2003? AB - Medical advances and the growth of the elderly population have focused interest on trends in the health of the elderly. Three theories have been advanced to describe these trends: compression of morbidity, expansion of morbidity, and dynamic equilibrium. We applied multistate life table methods to the Medicare Current Beneficiary Survey to estimate active and disabled life expectancy from 1992 to 2003, defining disability as having difficulty with instrumental activities of daily living or activities of daily living. We found increases in active life expectancy past age 65 and decreases in life expectancy with severe disability. These trends are consistent with elements of both the theory of compression of morbidity and the theory of dynamic equilibrium. PMID- 17913008 TI - The association of childhood socioeconomic conditions with healthy longevity at the oldest-old ages in China. AB - Based on unique data from the largest-ever sample of the Chinese oldest-old aged 80 and older, our multivariate logistic regression analyses show that either receiving adequate medical service during sickness in childhood or never/rarely suffering from serious illness during childhood significantly reduces the risk of being ADL (activities of daily living) impaired, being cognitively impaired, and self-reporting poor health by 18%-33% at the oldest-old ages. Estimates of effects for five other indicators of childhood conditions are similarly positive but mostly not statistically significant. Multivariate survival analysis shows that better childhood socioeconomic conditions in general tend to reduce the four year period mortality risk among the oldest-old. But after additional controls for 14 covariates are put into the model, the effects are not statistically significant, thus suggesting that most of the effects of childhood conditions on oldest-old mortality are indirect-at least to the point of affecting current health status at the oldest-old ages, which itself is strongly associated with mortality. While acknowledging limitations of the present analyses due to a lack of information on childhood illness, the oldest-olds'recollection errors, and other data problems, we conclude, based on this and other studies, that policies that enhance childhood health care and children's socioeconomic well-being can have large and long-lasting benefits up to the oldest-old ages. PMID- 17913010 TI - Race and the spatial segregation of jobless men in urban America. AB - Changes in U.S. metropolitan areas over the past 30 years are thought to have concentrated jobless men in low-income, predominantly minority neighborhoods clustered near the center of the city. Using tract-level data from the Neighborhood Change Database for 1970-2000, I examine how the residential segregation ofjobless from employed men has changed over the past three decades. I find that jobless men in U.S. metropolitan areas have become less uniformly distributed throughout the metropolis and more isolated, concentrated, and clustered since 1970; but they have also become less centralized. Racial and ethnic group differences in the spatial segregation of jobless men are large. Jobless black men occupy a uniquely disadvantaged ecological position in the metropolis: in comparison with other jobless men, they are much less uniformly distributed throughout the metropolis and much more isolated from employed men, they are concentrated in a smaller amount of physical space, and their neighborhoods are more clustered and are located closer to the center of the city. The dimensions of segregation strongly overlap for black jobless men, producing a multidimensional layering of segregation not encountered by other jobless men. Multivariate models reveal that the uniquely disadvantaged ecological position of jobless black men is less a reflection of different patterns of regional concentration and metropolitan settlement or of differences in group-status characteristics than it is an inevitable consequence of extreme levels of racial residential segregation in the United States. PMID- 17913009 TI - A fixed-effects multilevel analysis of how community family structure affects individual mortality in Norway. AB - By using register data for the entire Norwegian population aged 50-89 in the period 1980-1999, during which there were about 720,000 deaths, I estimate how the proportions of persons who were divorced or never married in the municipality affected all-cause mortality, net of individual marital status. The data include individual histories of changes in marital status and places of residence, providing a rare opportunity to enter municipality fixed effects into the model, thereby capturing the time-invariant unobserved factors at that level. The positive health externality of marriage that is suggested in the literature is supported by some of the estimates for women. Other estimates--especially those for men--point in the opposite direction. One possible interpretation of these findings is that social cohesion is perhaps not as beneficial for people's health as often claimed, at least not for both sexes. Alternatively, the results may reflect that marriage perhaps undermines rather than strengthens social cohesion, or that other mechanisms are involved-for example, those that are related to people 's perceptions of their health relative to the health of others. Estimates from models without such municipality fixed effects are markedly different, but these also shed doubt on the notion that a high proportion of unmarried persons generally increases individual mortality. PMID- 17913011 TI - National estimates of racial segregation in rural and small-town America. AB - The objective of this paper is to provide, for the first time, comparative estimates of racial residential segregation of blacks, Hispanics, and Native Americans in nonmetropolitan and metropolitan places in 1990 and 2000. Analyses are based on block data from the 1990 and 2000 U.S. decennial censuses. The results reveal a singularly important and perhaps surprising central conclusion: levels and trends in recent patterns of racial segregation in America's small towns are remarkably similar to patterns observed in larger metropolitan cities. Like their big-city counterparts, nonmetropolitan blacks are America's most highly segregated racial minority--roughly 30% to 40% higher than the indices observed for Hispanics and Native Americans. Finally, baseline ecological models of spatial patterns of rural segregation reveal estimates that largely support the conclusions reached in previous metropolitan studies. Racial residential segregation in rural places increases with growing minority percentage shares and is typically lower in "new" places (as measured by growth in the housing stock), while racially selective annexation and the implied "racial threat" at the periphery exacerbate racial segregation in rural places. Our study reinforces the need to broaden the spatial scale of segregation beyond its traditional focus on metropolitan cities or suburban places, especially as America's population shifts down the urban hierarchy into exurban places and small towns. PMID- 17913012 TI - Multipartnered fertility among American men. AB - Using the 2002 (Cycle 6) National Survey of Family Growth (NSFG), which was the first NSFG to interview men, we document the prevalence and correlates of sequential parenthood with different partners (multipartnered fertility) among a representative sample of American men. Nearly 8% of American men aged 15-44 report having had children with more than one partner, with sharp differences by age, race/ethnicity, and income-over one-third of poor black men aged 35-44 report having had children with two or more mothers, and 16% report children with three or more mothers. Fathers of two or more children by multiple partners appear to be more disadvantaged than fathers with two or more children by the same partner. Multipartnered fertility is strongly related to prior birth characteristics; men not in a coresidential union at the preceding birth are more likely to have their next birth with a new partner and controlling for prior birth characteristics accounts for the elevated risk of Hispanics and blacks in baseline models. Results also suggest that multipartnered fertility is becoming more prevalent as younger cohorts transition to a new-partner birth more quickly and at a higher rate than older cohorts. PMID- 17913013 TI - Contraceptive use patterns across teens' sexual relationships: the role of relationships, partners, and sexual histories. AB - By using data from the National Longitudinal Study of Adolescent Health, we examine how adolescent relationship characteristics, partner attributes, and sexual relationship histories are associated with contraceptive use and consistency, incorporating random effects to control for respondent-level unobserved heterogeneity. Analyses show that teens' contraceptive use patterns vary across relationships. Teens with more-homogamous partners, with more intimate relationships, and who communicate about contraception before sex have greater odds of contraceptive use and/or consistency. Teens in romantic relationships, and who are older when engaging in sex for the first time, have greater odds of ever using contraceptives but reduced odds of always using contraceptives. Teens continue habits from previous relationships: teens with experience practicing contraceptive consistency and females who previously have used hormonal contraceptive methods are better able to maintain consistency in subsequent relationships. Also, relationship and partner characteristics are less important for females who previously used hormonal methods. PMID- 17913014 TI - How do marital status, work effort, and wage rates interact? AB - How marital status interacts with men's earnings is an important analytic and policy issue, especially in the context of debates in the United States over programs that encourage healthy marriage. This paper generates new findings about the earnings-marriage relationship by estimating the linkages among flows into and out of marriage, work effort, and wage rates. The estimates are based on National Longitudinal Survey of Youth panel data, covering 23 years of marital and labor market outcomes, and control for unobserved heterogeneity. We estimate marriage effects on hours worked (our proxy for work effort) and on wage rates for all men and for black and low-skilled men separately. The estimates reveal that entering marriage raises hours worked quickly and substantially but that marriage's effect on wage rates takes place more slowly while men continue in marriage. Together; the stimulus to hours worked and wage rates generates an 18% 19% increase in earnings, with about one-third to one-half of the marriage earnings premium attributable to higher work effort. At the same time, higher wage rates and hours worked encourage men to marry and to stay married. Thus, being married and having high earnings reinforce each other over time. PMID- 17913015 TI - Self-selection and earnings assimilation: immigrants from the former Soviet Union in Israel and the United States. AB - Drawing on U.S. decennial census data and on Israeli census and longitudinal data, we compare the educational levels and earnings assimilation of Jewish immigrants from the former Soviet Union (FSU) in the United States and Israel during 1968-2000. Because the doors to both countries were practically open to FSU immigrants between 1968 and 1989, when FSU immigrants were entitled to refugee visas in the United States, the comparison can be viewed as a natural experiment in immigrants' destination choices. The results suggest that FSU immigrants to the United States are of significantly higher educational level and experience significantly faster rates of earnings assimilation in their new destination than their counterparts who immigrated to Israel. We present evidence that patterns of self-selection in immigration to Israel and the United States- on both measured and unmeasured productivity-related traits--is the main reason for these results. When the immigration regulations in the United States changed in 1989, and FSU Jewish immigrants to the United States had to rely on family reunification for obtaining immigrant visas, the adverse effects of the policy change on the type of FSU immigrants coming to the United States were minor and short-lived As early as 1992, the gaps in the educational levels between FSU immigrants coming to Israel and to the United States returned to their pre-1989 levels, and the differences in earnings assimilation of post-1989 immigrants in the United States and Israel are similar to the differences detected in the 1980s. PMID- 17913016 TI - The political ecology of disaster: an analysis of factors influencing U.S. tornado fatalities and injuries, 1998-2000. AB - This study examines casualties from tornadoes in the United States between the years 1998 and 2000. A political model of human ecology (POET) was used to explore how the environment, technology, and social inequality influence rates of fatalities and injuries in two models. Data were drawn from four sources: John Hart's Severe Plot v2.0, National Weather Service (NWS) Warning Verification data, Storm Prediction Center (SPC) watch data, and tract-level census data. Negative binomial regression was used to analyze the causes of tornado fatalities and injuries. Independent variables (following POET) are classified in the following manner: population, organization, environment, and technology. Rural population, population density, and household size correspond to population; racial minorities and deprivation represent social organization; tornado area represents environment; and tornado watches and warnings, as well as mobile homes, correspond to technology. Findings suggest a strong relationship between the size of a tornado path and both fatalities and injuries, whereas other measures related to technology, population, and organization produce significant yet mixed results. Census tracts having larger populations of rural residents was, of the nonenvironmental factors, the most conclusive regarding its effects across the two models. The outcomes of analysis, although not entirely supportive of the model presented in this study, suggest to some degree that demographic and social factors play a role in vulnerability to tornadoes. PMID- 17913017 TI - Beauty in chaos. PMID- 17913018 TI - Machine learning to boost the next generation of visualization technology. PMID- 17913019 TI - Action- and workflow-driven augmented reality for computer-aided medical procedures. PMID- 17913020 TI - Discovering the unexpected. PMID- 17913021 TI - Visual discovery in computer network defense. PMID- 17913022 TI - Visual insights into high-resolution earthquake Simulations. PMID- 17913023 TI - Visualizing diversity and depth over a set of objects. PMID- 17913024 TI - nSpace and GeoTime: a VAST 2006 case study. PMID- 17913025 TI - Bridging the semantic gap: visualizing transition graphs with user-defined diagrams. PMID- 17913026 TI - Occlusion-resistant camera design for acquiring active environments. PMID- 17913027 TI - Retargeting images and video for preserving information saliency. PMID- 17913028 TI - Envisioning the wind: meteorology graphics at weather underground. PMID- 17913029 TI - Digital art: when artistic and cultural muse merges with computer technology. PMID- 17913030 TI - Let's hear it for the good news. PMID- 17913031 TI - Turning a blind eye. PMID- 17913032 TI - NICE judgement leaves behind a nasty taste. PMID- 17913033 TI - Managing risks. PMID- 17913034 TI - How well are admission avoidance schemes working and do they increase the risk of denying older people access to acute hospital services if they need them? PMID- 17913035 TI - Letting services flourish. PMID- 17913036 TI - Are you at risk? PMID- 17913037 TI - Cafe society. PMID- 17913038 TI - Otherwise engaged. PMID- 17913039 TI - Helping patients to cope with faecal incontinence. PMID- 17913040 TI - Community matrons: promoting self management (part four). AB - This is the fourth of six articles which draw on the content of a series of masterclasses organised and funded by the Department of Health for community matrons during summer 2006. It draws on input to the community matrons masterclasses from Keith Strahan and Alf Collins. It opens by reviewing the concepts of self care and self management. The significance of understnading patients' and their carers' health beliefs in person-centred care planning is then highlighted. The importance of capitalising on readiness to change the role of motivational interviewing in successful behaviour change is then presented. Finally, the consequent issues for professionals and services in terms of managing risk are discussed. PMID- 17913041 TI - Assessing falls in older people. AB - For an older person in hospital a fall can have devastating consequences, prolonging hospital stay and leading to loss of confidence. Falls cost the NHS between pounds 5340 and pounds 12,500 per patient in delayed discharge, and can cause nurses to question their own practice and blame themselves. Further, the efforts of clinicians and researchers implementing falls prevention interventions are hampered because of inconsistencies in assessing risk. The use of falls risk assessment tools varies throughout the U.K. In Scotland some hospitals use them while others do not. Unlike England and Wales, Scotland has no national guidelines. To deal with this problem, the authors conclude that it is important to develop an evidence-based guideline that covers the whole of the UK. PMID- 17913042 TI - Elizabeth Atere-Roberts. PMID- 17913043 TI - TAT peptide-modified liposomes provide enhanced gene delivery to intracranial human brain tumor xenografts in nude mice. AB - In this study, we have investigated the potential of trans-activating transcriptional activator peptide (TATp)-modified liposomes to enhance the delivery of the model gene, plasmid encoding for the green fluorescent protein (pEGFP-N1), to human brain tumor U-87 MG cells in vitro and in an intracranial model in nude mice. The TATp-lipoplexes were characterized at lipid/DNA (+/-) charge ratios of 0.2, 5, 10, and 20 for size analysis and DNA complexation. The size distribution of DNA-loaded TATp-liposomes was narrow and the DNA complexation was firm at lipid/DNA (+/-) charge ratios of 5 and higher. TATp lipoplexes had demonstrated an enhanced delivery of pEGFP-N1 to U-87 MG tumor cells in vitro at lipid/DNA (+/-) charge ratios of 5 and 10. In vivo transfection of intracranial brain tumors by intratumoral injections of TATp-lipoplexes showed an enhanced delivery of pEGFP-N1 selectively to tumor cells and subsequent effective transfection compared to plain plasmid-loaded lipoplexes. No transfection (green fluorescence of the GFP) was noted in the normal brain adjacent to tumor. PMID- 17913044 TI - Increased preclinical efficacy of irinotecan and floxuridine coencapsulated inside liposomes is associated with tumor delivery of synergistic drug ratios. AB - Whether anticancer drug combinations act synergistically or antagonistically often depends on the ratio of the agents being combined. We show here that combinations of irinotecan and floxuridine exhibit drug ratio-dependent cytotoxicity in a broad panel of tumor cell lines in vitro where a 1:1 molar ratio consistently provided synergy and avoided antagonism. In vivo delivery of irinotecan and floxuridine coencapsulated inside liposomes at the synergistic 1:1 molar ratio (referred to as CPX-1) lead to greatly enhanced efficacy compared to the two drugs administered as a saline-based cocktail in a number of human xenograft and murine tumor models. When compared to liposomal irinotecan or liposomal floxuridine, the therapeutic activity of CPX-1 in vivo was not only superior to the individual liposomal agents, but the extent of tumor growth inhibition was greater than that predicted for combining the activities of the individual agents. In contrast, liposome delivery of irinotecan:floxuridine ratios shown to be antagonistic in vitro provided antitumor activity that was actually less than that achieved with liposomal irinotecan alone, indicative of in vivo antagonism. Synergistic antitumor activity observed for CPX-1 was associated with maintenance of the 1:1 irinotecan:floxuridine molar ratio in plasma and tumor tissue over 16-24 h. In contrast, injection of the drugs combined in saline resulted in irinotecan:floxuridine ratios that changed 10-fold within 1 h in plasma and sevenfold within 4 h in tumor tissue. These results indicate that substantial improvements in the efficacy of drug combinations may be achieved by maintaining in vitro-identified synergistic drug ratios after systemic administration using drug delivery vehicles. PMID- 17913045 TI - IL-2, TNF-alpha, and leptin: local versus systemic concentrations in NSCLC patients. AB - One recent line of cancer research shows increasing interest for biological factor such as IL-2, TNF-alpha, and leptin, which have been found to participate in the development and progression of non-small cell lung cancer (NSCLC). The aim of this study was to measure IL-2, TNF-alpha, and leptin concentrations in the airways and in the systemic circle of patients with NSCLC, investigating the role of these factors in the lung tumors. We enrolled 32 patients (17 men, 71 +/- 7 years) with a histological diagnosis of NSCLC and 20 healthy ex-smoker controls, negative for computed tomography of the chest (14 men, 69 +/- 8 years). IL-2, TNF alpha, and leptin levels were measured in the serum, the urine, the bronchoalveolar lavage, the induced sputum, and exhaled breath condensate (EBC) of patients enrolled by means of a specific enzyme immunoassay kit. Higher concentrations of IL-2, TNF-alpha and leptin were found in NSCLC patients than in controls (p < 0.0001). A statistically significant increase of IL-2, TNF-alpha, and leptin concentrations was observed in patients from stage I to stage III of NSCLC. These findings suggest that IL-2, TNF-alpha, and the leptin play an important role in the cancerogenesis of NSCLC. Their measure in the EBC could be proposed as noninvasive markers for an early detection of NSCLC and in the follow up of this tumor. PMID- 17913046 TI - Prenatal and perinatal risk factors and testicular cancer: a hospital-based case control study. AB - Some evidence exists to support the hypothesis that elevated levels of circulating maternal estrogens during early pregnancy may increase risk of testicular germ cell cancer. However, the results from studies evaluating maternal factors have been mixed. We evaluated maternal factors, particularly those associated with excess estrogen levels, as risk factors for testicular cancer. We conducted a hospital-based case-control study at The University of Texas M. D. Anderson Cancer Center in Houston, Texas of 144 testicular cancer patients diagnosed between 1990 and 1996 and 86 friend controls matched to cases on age, race, and state of residence. Risk factor data about the mother, the son, and the pregnancy were obtained from the mothers by telephone interviews and from the sons by self-administered questionnaires. Extreme nausea during the first trimester of pregnancy was associated with an elevated risk of testicular cancer [odds ratio (OR) = 2.0; 95% confidence interval (CI) = 1.0-3.9]. Adjustment for potential confounders slightly lowered this risk (OR = 1.8; 95% CI = 0.9-3.8). Risks were modestly increased for other factors that are proxy measures for maternal estrogens, including preterm delivery (OR = 2.2; 95% CI = 0.4-12.9), birth weight <3000 g (OR = 2.4: 95% CI = 0.7-8.1), and birth weight >4000 g (OR = 1.7; 95% CI = 0.9-3.2), albeit nonsignificantly so. Our finding that severe nausea was associated with increased testicular cancer risk adds evidence to support the in utero estrogen exposure hypothesis because nausea early in pregnancy is related to rising levels of circulating estrogens. For other factors, which are less direct measures of maternal estrogens, the modest associations found indicate a suggestive pattern in support of the excess estrogen hypothesis. PMID- 17913047 TI - Radiation-induced antitumor properties of vitamin B5 (pantothenic acid) and its effect on mitomycin C activity: experiments in vitro. AB - Vitamin B5 (pantothenic acid) shows a strongly pronounced antitumor effect under the influence of ionizing radiation. In the frame of experiments in vitro (model: Escherichia coli bacteria, AB1157) performed under the exact knowledge of concentration and kind of the free radicals acting in the various aqueous media (pH 7.4) the following was established: (i) vitamin B5 possesses a very intense antitumor property, (ii) it exerts a strong synergistic effect on mitomycin C (MMC), (iii) the oxidizing species (OH*, O2*-) appears to be most important in the initiation of the observed effect. The generated radiolytic products from vitamin B5 very likely also play an important role in this respect. PMID- 17913048 TI - Expression of MDR1 in epithelial ovarian cancer and its association with disease progression. AB - The purposes of this study were to analyze MDR1 expression in ovarian tumors prior to chemotherapy, to correlate the expression with p16, IGFs, ERalpha, and BRCA1, and to examine the association of MDR1 expression with ovarian cancer prognosis. A primary ovarian cancer cohort of 206 patients after surgery was followed up. MDR1, IGFs, ERalpha, p16, and BRCA1 expressions were analyzed in ovarian tumor samples using quantitative real-time PCR. MDR1 was detected in 177 of 206 specimens. MDR1 expression was positively correlated with IGFBP3, ERalpha, p16, and BRCA1, but not correlated with IGF-II, age, and other clinicopathological parameters. MDR1 expression significantly elevated the risk for disease progression (p = 0.02), and this association remained statistically significant after controlling for patient age and clinicopathological parameters or other correlated genes. No association was found between MDR1 expression and overall survival. MDR1 expression may be an independent marker for ovarian cancer progression and combination of different agents targeting different molecules may improve the outcome of ovarian cancer treatment and prevent drug resistance. PMID- 17913049 TI - [Foot and mouth disease is coming back]. PMID- 17913050 TI - [Bat research on rabies virus in the Netherlands from 2002 to 2006]. PMID- 17913051 TI - [Part 5. Single cell parasites (ciliates and flagellates) and multiple cell parasites (Trematoda)]. PMID- 17913052 TI - [Treatment relations]. PMID- 17913054 TI - [Argos, number 36, spring 2007]. PMID- 17913053 TI - [New research in preventive medicine]. PMID- 17913055 TI - [Mammae health panel spring 2007]. PMID- 17913056 TI - [Wiser in animal welfare]. PMID- 17913057 TI - [Work meetings nationally coordinated. Interview by Henry Dijkman]. PMID- 17913058 TI - Pyrrolizidine alkaloid profiles of the Senecio cineraria group (Asteraceae). AB - Alkaloid profiles of five Senecio species (Asteraceae), including S. ambiguus subsp. ambiguus, S. ambiguus subsp. nebrodensis, S. gibbosus subsp. bicolor, S. gibbosus subsp. gibbosus, and S. gibbosus subsp. cineraria, were studied. Eleven pyrrolizidine alkaloids were identified and their content was evaluated by GLC-MS and GLC analysis. Otosenine and florosenine were found to be the major alkaloids in all studied species. It is interesting that only S. ambiguus subsp. nebrodensis was characterized by a high content of the alkaloids jacobine, jacoline, jaconine, and jacozine. PMID- 17913059 TI - Variabilin, a chemotaxonomic marker for the family Irciniidae. AB - The furanosesterterpene variabilin was isolated from the sponge Sarcotragus. From a chemical point of view, the family Irciniidae has been the source of furanosesterterpenes, and especially variabilin is an important chemotaxonomic marker for the family Irciniidae. PMID- 17913060 TI - Preliminary evaluation of antimicrobial activity of diastereomeric cis/trans-3 aryl(heteroaryl)-3,4-dihydroisocoumarin-4-carboxylic acids. AB - Preliminary differentiating screening of the antibacterial and antifungal activity of a series of diastereomeric cis/trans-3-aryl(heteroaryl)-3,4 dihydroisocoumarin-4-carboxylic acids (3a-i) was performed by the agar diffusion method against twelve microorganism strains of different taxonomic groups. S. aureus and A. niger were the most sensitive strains to the antibiotic effect of the tested compounds, both inhibited by 10 of 12 compounds. The most potent antibacterial agent was cis-3-phenyl-3,4-dihydroisocoumarin-4-carboxylic acid (cis-3a), exhibiting activity against all seven bacterial test strains. PMID- 17913061 TI - Carbohydrazones of substituted salicylaldehydes as potential lead compounds for the development of narrow-spectrum antimicrobials. AB - Certain substituted salicylaldehydes are known to have highly potent antimicrobial activity against bacteria and fungi, but the mechanism underlying this remarkable activity is not known, and almost nothing has been reported on the effects of further modification of the structures, such as the formation of hydrazone-type derivatives. We report now a study on the antimicrobial properties of the carbohydrazone derivatives of several substituted salicylaldehydes. The compounds studied were synthesized from ring-substituted salicylaldehydes and carbohydrazide in the mole ratio 2:1. They were tested against Aspergillus niger, Bacillus cereus, Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Saccharomyces cerevisiae and Staphylococcus epidermidis using the agar diffusion method. The carbohydrazone derived from 2,3,4-trihydroxybenzaldehyde had distinctly higher activity than the parent aldehyde in the same molar concentration. This activity was limited to one test organism (S. epidermidis), while the free aldehyde had at least some (in some cases even high) activity against all of the microbes studied. All other ones of the effective carbohydrazone compounds were distinctly less active than the parent salicylaldehydes as such. The hydrazones studied had in general a narrower antimicrobial spectrum than the free aldehydes and are thus of interest as potential lead compounds for the development of narrow-spectrum anti-microbial drugs. The mechanism of action of the aldehydes as well as that of the carbohydrazones is discussed. PMID- 17913062 TI - Antimicrobial properties of substituted salicylaldehydes and related compounds. AB - A systematic survey of the antimicrobial properties of substituted salicylaldehydes and some related aromatic aldehydes is reported. A total of 23 different compounds, each at four different concentrations, were studied using a panel of seven microbes (Aspergillus niger, Bacillus cereus, Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Saccharomyces cerevisiae and Staphylococcus epidermidis) and employing the paper disc agar diffusion method. Several aldehydes, most notably halogenated, nitro-substituted and hydroxylated salicylaldehydes, displayed highly potent activity against the microbes studied, giving inhibitory zones up to 49 mm in diameter (paper disc diameter 6 mm), while unsubstituted benzaldehyde and salicylaldehyde had minimal activity. Further, 4,6 dimethoxysalicylaldehyde had considerable activity against C albicans and slight activity against S. cerevisiae, while displaying minimal activity against bacteria. Also two aromatic dialdehydes had interesting activity. In general, P. aeruginosa was the least sensitive microbe, a result that is in line with observations from a large screening project, in which this microbe was the one against which the least number of active substances was found. Interestingly, the structure-activity relationships of the aldehydes studied were clearly different for different microbes. Many of the aldehydes tested had such high antimicrobial activity that they are noteworthy candidates for practical applications as well as interesting lead compounds for the development of novel antimicrobial drugs and disinfectants. The structure-activity relationships are discussed in detail. For high activity, substituents are required in benzaldehyde as well as in its 2 hydroxy derivative salicylaldehyde. One hydroxy group alone (at the 2-position) is not enough, but further hydroxylation may produce high activity. The effects of substituents are in some cases dramatic. Halogenation, hydroxylation and nitro substitution may produce highly active compounds, but the effects are not easily predicted nor can they be extrapolated from one microbe to another. PMID- 17913063 TI - Antimicrobial activity of two antitumour agents and ribonucleotide reductase inhibitors, pyridine-2-carboxaldehyde thiosemicarbazone and the acetate form of its copper(II) chelate. AB - Some copper chelates have potent antitumour activity, and in some cases also the free ligands have activity in vivo. Yet, little is known about their antimicrobial properties. Copper(II) chelates of the thiosemicarbazones of a-N heterocyclic carboxaldehydes constitute one important group of such agents, also their ligands having marked antitumour activity. Both the ligands and chelates inhibit ribonucleotide reductase. Some ligands have been or are under clinical trials as antineoplastic agents. I report here a study on the antimicrobial properties of the prototype compounds of this group, pyridine-2-carboxaldehyde thiosemicarbazone and its copper(II) chelate. They were tested against nine microbes, including bacteria (Bacillus cereus, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus lactis), yeasts (Candida albicans and Saccharomyces cerevisiae) and one mold (Aspergillus niger). Two clinical isolates of Bacillus sp. and one reference strain were also studied. Both the ligand and the chelate had marked activity. The ligand displayed considerable activity against all bacteria except for S. lactis, and its activity against E. coli and P. aeruginosa was that high that practical applications might be considued. It was highly active against A. niger and moderately active against C. albicans. The chelate was highly active against S. epidermidis and S. cerevisiae. Both compounds inhibited the clinical isolates markedly. Since some related ligands have been or are in clinical trials on humans or are entering them, their route to clinical use, also as antimicrobials, might be much more straightforward than that of substances, whose toxicity in humans is wholly unexplored. PMID- 17913064 TI - Screening of antibacterial activities of twenty-one oxygenated monoterpenes. AB - Plant essential oils are widely used as fragrances and flavours in the cosmetic, perfume, drug and food industries. Oxygenated monoterpenes are widespread components of the essential oils, usually occurring in high amount. In this paper, the antibacterial activities of twenty-one oxygenated monoterpenes (borneol, borneol acetate, camphor, carvone, 1,8-cineole, citronellal, beta citronellol, dihydrocarvone, fenchol, fenchone, geraniol acetate, isomenthol, limonene oxide, linalool, linalool acetate, nerol, nerol acetate, terpinen-4-ol, alpha-terpineol, menthol and menthone) and penicillin (standard antibiotic) were determined using a disc diffusion method (in vitro) against 63 bacterial strains, belonging to 37 different genera and 54 species (plant, food and clinic origins). The results showed that the oxygenated monoterpenes exhibited a variable degree of antibacterial activities. These compounds also inhibited the growth of bacterial strains by producing a weak zone of inhibition from 7 to 11 mm in diameter, depending on the susceptibility of the tested bacteria. Among the tested compounds, nerol, linalool alpha-terpineol, fenchol and terpinen-4-ol showed antibacterial activity at a broad spectrum. However, their antibacterial activities were lower than those of penicillin. In contrast to these compounds, camphor and 1,8-cineole exhibited no inhibition effects on the growth of all tested bacteria. PMID- 17913065 TI - Antimicrobial activity of some Salvia species essential oils from Iran. AB - The aerial parts of Salvia multicaulis, S. sclarea and S. verticillata were collected at full flowering stage. The essential oils were isolated by hydrodistillation and analyzed by combination of capillary GC and GC-MS. The in vitro antimicrobial activity of the essential oils were studied against eight Gram-positive and Gram-negative bacteria (Bacillus subtilis, Bacillus pumulis, Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae) and three fungi (Candida albicans, Saccharomyces cerevisiae and Aspergillus niger). The results of antibacterial activity tests of the essential oils according to the disc diffusion method and MIC values indicated that all the samples have moderate to high inhibitory activity against the tested bacteria except for P. aeruginosa which was totally resistant. In contrast to antibacterial activity, the oils exhibited no or slight antifungal property, in which only the oil of S. multicaulis showed weak activity against two tested yeasts, C. albicans and S. cerevisiae. PMID- 17913066 TI - In vivo anti-inflammatory and antinociceptive activity evaluation of phenolic compounds from Sideritis stricta. AB - An acetone extract obtained from aerial parts of S. stricta Boiss. & Heldr. apud Bentham, its fractions and phenolic compounds were investigated for their in vivo anti-inflammatory and antinociceptive activities. For the anti-inflammatory activity and for the antinociceptive activity assessment, carrageenan-induced hind paw edema and p-benzoquinone-induced abdominal constriction tests were used, respectively. The acetone extract of the plant and its phenolic fraction exhibited potent inhibitory activity against both bioassay models in mice. From the active phenolic fraction a well-known phenylethanoid glycoside, verbascoside (acteoside) (1), and two flavonoid glycosides, isoscutellarein 7-O-[6"'-O-acetyl beta-D-allopyranosyl-(1-->2)]-beta-D-glucopyranoside (2) and isoscutellarein 7-O [6"'-O-acetyl-beta-D-allopyranosyl-(1-->2)]-6"-O-acetyl-beta-D-glucopyranoside (3), were isolated. During phytochemical studies we also isolated a methoxyflavone, xanthomicrol (4), from the non-polar fraction. The structures of the isolated compounds were established by spectroscopic evidence (UV, IR, 1D- and 2D-NMR, MS). Although antinociceptive and anti-inflammatory activities of the phenolic components were found not significant in the statistical analysis, compounds 1 to 3 showed a notable activity without inducing any apparent acute toxicity as well as gastric damage. Furthermore, a mixture of flavonoid glycosides (2 + 3) exhibited a significant inhibitory effect in both models at a higher dose. PMID- 17913067 TI - Anticancer and antioxidant tannins from Pimenta dioica leaves. AB - Two galloylglucosides, 6-hydroxy-eugenol 4-O-(6'-O-galloyl)-beta-D-4C1 glucopyranoside (4) and 3-(4-hydroxy-3-methoxyphenyl)-propane-1,2-diol-2-O-(2',6' di-O-galloyl)-beta-D -4C1-glucopyranoside (7), and two C-glycosidic tannins, vascalaginone (10) and grandininol (14), together with fourteen known metabolites, gallic acid (1), methyl gallate (2), nilocitin (3), 1-O-galloyl-4,6 (S)-hexahydroxydiphenoyl-(alpha/beta)-D-glucopyranose (5), 4,6-(S) hexahydroxydiphenoyl-(alpha/beta)-D-glucopyranose (6), 3,4,6-valoneoyl (alpha/beta)-D-glucopyranose (8), pedunculagin (9), casuariin (11), castalagin (12), vascalagin (13), casuarinin (15), grandinin (16), methyl-flavogallonate (17) and ellagic acid (18), were identified from the leaves of Pimenta dioica (Merr.) L. (Myrtaceae) on the basis of their chemical and physicochemical analysis (UV, HRESI-MS, 1D and 2D NMR). It was found that 9 is the most cytotoxic compound against solid tumour cancer cells, the most potent scavenger against the artificial radical DPPH and physiological radicals including ROO*, OH*, and O2-*, and strongly inhibited the NO generation and induced the proliferation of T lymphocytes and macrophages. On the other hand, 3 was the strongest NO inhibitor and 16 the highest stimulator for the proliferation of T-lymphocytes, while 10 was the most active inducer of macrophage proliferation. PMID- 17913068 TI - Nerolidol, an antiulcer constituent from the essential oil of Baccharis dracunculifolia DC (Asteraceae). AB - In this study, the antiulcerogenic effect of essential oil from Baccharis dracunculifolia was evaluated using the model of acute gastric lesions induced by ethanol. The ulcerative lesion index (ULI) was significantly reduced by oral administration of the essential oil of B. dracunculifolia at doses of 50, 250 and 500 mg/kg which reduced the lesions by 42.79, 45.70 and 61.61%, respectively. The analysis of the chemical composition of the essential oil from B. dracunculifolia by GC showed that this was composed mainly of mono- and sesquiterpenes and the majority compound was nerolidol. Therefore, antiulcerogenic activity of nerolidol (50, 250 and 500 mg/kg) was investigated using ethanol-, indomethacin- and stress induced ulcer models in rat. In the stress-induced ulcer model, a significant reduction of the ULI in animals treated with nerolidol (50, 250 and 500 mg/kg) and cimetidine (100 mg/kg) was observed, compared to the control group (p < 0.05). The percentage of inhibition of ulcer was 41.22, 51.31, 56.57 and 53.50% in groups treated with 50, 250, 500 mg/kg of nerolidol and 100 mg/kg of cimetidine (positive control), respectively. Regarding ethanol- and indomethacin induced ulcer models, it was observed that the treatment with nerolidol (250 and 500 mg/ kg) significantly reduced the ULI in comparison with the control group (p < 0.05). The dose of 50 mg/kg reduced the parameters analyzed but this was not statistically significant. In the ethanol-induced model percentage of inhibition of ulcer was 34.20, 52.63, 87.63 and 50.87% in groups treated with 50, 250, 500 mg/kg of nerolidol and 30 mg/kg of omeprazol (positive control), respectively. In indomethacin-ulcer the percentage of inhibition of ulcer was 34.69, 40.80, 51.02 and 46.93% in groups treated with 50, 250, 500 mg/kg of nerolidol and 100 mg/ kg of cimetidine (positive control), respectively. The results of this study show that nerolidol displays antiulcer activity, as it significantly inhibited the formation of ulcers induced in different animal models. However, further pharmacological and toxicological investigations, to delineate the mechanism(s) of action and the toxic effects, are required to allow the use of nerolidol for the treatment of gastric ulcer. PMID- 17913069 TI - Lichen secondary metabolites from the cultured lichen mycobionts of Teloschistes chrysophthalmus and Ramalina celastri and their antiviral activities. AB - Lichens and spore-derived cultured mycobionts of Teloschistes chrysophthalmus and Ramalina celastri were studied chemically, and results indicated that they produced, respectively, parietin and usnic acid as major secondary metabolites, which were purified and identified. Identification of the compounds was performed by high performance liquid chromatography and structural elucidation by nuclear magnetic resonance (1H) and electron impact mass spectrometry. Usnic acid exhibited antiviral activity whereas parietin had a virucidal effect against the arenaviruses Junin and Tacaribe. PMID- 17913070 TI - Phytochemical and antinociceptive properties of Matayba elaeagnoides Radlk. barks. AB - A mixture of triterpenes named lupeol (1), alpha-amyrin (2), beta-amyrin (3), and beta-sitosterol (4) has been isolated from the hexane fraction of Matayba elaeagnoides. In addition, scopoletin (5), umbelliferone (6), 3beta-O-D glycopyranosyl-sitosterol (7) and betulin (8) were isolated from the chloroform fraction. All the structures were identified by spectroscopic techniques in accordance with literature data. The extracts (hydroalcoholic and methanolic) and some fractions (hexane, chloroform, ethyl acetate and butanol) exerted promising antinociceptive effects in mice. In addition, we have tested the pure compound betulin (8). When analyzed against induced pain using the writhing test (3-10 mg kg(-1), i.p.), betulin showed a dose-dependent effect with a calculated ID50 value of 7.74 (6.53-9.17) mg kg(-1) [17.5 (14.7-20.7) micromol kg(-1)] and a maximal inhibition (MI) of 58.3% in relation to the control group. When evaluated in the formalin test (3-10 mg kg(-1), i.p.), this compound inhibited both phases of pain (neurogenic and inflammatory pain), with calculated ID50 values of 18.3 (17.7-18.9) and 8.3 (7.7-8.9) mg kg(-1) [41.5 (38.4-42.7) and 18.8 (17.6-19.9) micromol kg(-1)] and maximal inhibition of 40.8 and 64.39% for the first and second phases, respectively. Using the same models, this compound was several times more active than two clinically used drugs, namely aspirin and paracetamol, suggesting that its main active principle is related to the antinociceptive effect found for the chloroform fraction of M. elaeagnoids barks. PMID- 17913071 TI - Free radical scavenging activity and secondary metabolites from in vitro cultures of Sanicula graveolens. AB - An in vitro propagation system was developed to obtain shoot and root cultures from the Andean spice Sanicula graveolens (Apiaceae). Propagation of shoots, roots and plantlets was achieved by the temporary immersion system. The free radical scavenging effect of the methanol/water (7:3 v/v) extracts was determined by the discoloration of the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH). Total phenolic, flavonoid, chlorogenic acid (CA) and quercetin 3-O-glucoside content in the samples was assessed by spectrophotometry and DAD-HPLC analysis, respectively. On a dry weight basis, the crude extracts showed total phenolic values ranging from 3.57 to 6.93%, with highest content for the root culture sample. Total flavonoid content ranged from 1.23 to 2.23% and was lower for the root culture. Chlorogenic acid and neochlorogenic acid were identified by TLC in all samples. Highest free radical scavenging effect was observed for the root culture which also presented the highest CA content. Two of the shoot culture samples, with similar IC50 values in the DPPH discoloration assay, also presented close quercetin-3-O-glucoside content. PMID- 17913072 TI - Sedative effect of monoterpene alcohols in mice: a preliminary screening. AB - Many essential oils and monoterpenes are used therapeutically as relaxing drugs and tranquilizers. In this study, ten structurally related monoterpene alcohols, present in many essential oils, were evaluated in mice to investigate their pharmacological potential in the central nervous system. Isopulegol (1), neoisopulegol (2), (+/-)-isopinocampheol (3), (-)-myrtenol (4), (-)-cis-myrtanol (5), (+)-p-menth-1-en-9-ol (6) and (+/-)-neomenthol (8) exhibited a depressant effect in the pentobarbital-induced sleep test, indicating a sedative property. ( )-Menthol (7), (+)-dihydrocarveol (9), and (+/-)-isoborneol (10) were ineffective in this test. The results show that these psychoactive monoterpenes have the profile of sedative drugs, and this pharmacological effect is influenced by the structural characteristics of the molecules. PMID- 17913073 TI - Mollisianitrile, a new antibiotic from Mollisia sp. A59-96. AB - Mollisianitrile (1), a new antibiotic was isolated from the fermentation broth of Mollisa sp. A59-96 together with the two known isocoumarins 2 and 3. 1 exhibited antimicrobial, cytotoxic, and phytotoxic activities. 1 contains a reactive propiolonitrile moiety which is believed to be responsible for its antibiotic activities. Upon incubation with L-cysteine the biological activity was lost. PMID- 17913074 TI - Physiological versatility of the genus Rhodocista. AB - A new purple bacterium (strain T4), capable of heterotrophic aerobic and phototrophic anaerobic growth, was isolated from waste water of a noodle factory near Hanoi, Vietnam. A comparison of 16S rDNA sequences revealed its association with the genus Rhodocista. The isolate, tentatively named "Rhodocista hanoiensis", forms cysts after growth on butyrate-containing plates at 42 degrees C. The vegetative cells form short (under aerobic conditions) or long curve shaped rods. In contrast to other species of this genus T4 does not need any supplines (growth factors, not synthesized by the organisms). Comparative studies of T4 with Rhodocista centenaria (Rhodospirillum centenum) and Rhodocista pekingensis revealed a remarkable physiological versatility regarding nutrient spectra and survival properties of this genus. PMID- 17913075 TI - Production of exopolysaccharides by a submerged culture of an entomopathogenic fungus, Paecilomyces sp. AB - Exopolysaccharide basic was obtained from a submerged culture of a native Paecilomyces sp. strain isolated from Chilean soil. PMID- 17913076 TI - Periploca sepium Bunge as a model plant for rubber biosynthesis study. AB - Periploca sepium Bunge (Chinese silk vine) is a woody climbing vine belonging to the family Asclepiadaceae. It originally comes from Northwest China. Periploca resembles the Para-rubber tree, Hevea brasiliensis, regarding a similar body plan to produce a milky exudate containing rubber latex. The Periploca plant was assessed as a rubber-producing plant by rubber structure elucidation and its molecular weight distribution. A rubber fraction purified from the milky exudate was subjected to 1H NMR analysis, and a characteristic signal derived from cis polyisoprene was observed. In addition, when the molecular weight distribution of rubber components in the exudate was measured (using size-exclusion chromatography), the number-average molecular weight (Mn), weight-average molecular weight (Mw), and polydispersity (Mw/Mn) were estimated to be Mn = 1.3 x 10(5), Mw = 4.1 x 10(5), and Mw/Mn = 3.1, respectively. Furthermore, the presence of polyisoprene, with Mn = 4.0 x 10(4), Mw = 7.6 x 10(4), and Mw/Mn = 2.5, was also confirmed in plantlets obtained from shoots as a result of tissue culture. PMID- 17913077 TI - Transgenic rice plants expressing a novel antifreeze glycopeptide possess resistance to cold and disease. AB - Freezing injury and disease are both restrictive factors in crop production. In order to improve the tolerance ability to these stresses, a better way is to carry out genetic engineering by transferring dualfunctional genes. A predicted rice antifreeze glycopeptide gene was purposefully selected from rice blast induced cDNA library. Northern blot demonstrated that the gene is expressed not only in blast-infected rice leaves, but also in low temperature-treated rice. In addition, the expressed protein in Escherichia coli exhibits strong antifreeze activities. The gene was overexpressed in rice plants transformed via Agrobacterium tumefacient EHA105. Overall 112 T0 transformants were obtained in this research. Cold tolerance and disease resistance of T1 transformants were, respectively, investigated. The results showed that plants containing overexpressed transgene can withstand -1 degrees C for 24 h without severe chilling injury after thawed, and that disease symptoms of the parallel transformants are highly reduced in response to blast infection, when compared with controls. The relationship of the gene and several pathogenesis-related protein genes to be chosen was analyzed and discussed. All these results confirmed the dual role of the cloned gene, and implied that genetic engineering using this kind of gene is a promising method to reduce biotic and abiotic stresses. PMID- 17913078 TI - Ischemia-responsive protein (irp94) gene expression in neurons. AB - An increased expression of the ischemia-responsive protein gene (irp94) was detected in a Mongolian gerbil brain after an ischemic injury, particularly in the cerebral cortex and hippocampus. In a rat phaeochromocytoma tumour cell line (PC12 cells), actinomycin D blocked the irp94 gene expression but cycloheximide did not. This indicates that irp94 gene expression is transcriptionally controlled. The half-life of irp94 mRNA was estimated to be approx. 5 h using 5,6 dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB). In addition, irp94 expression was enhanced by either endoplasmic reticulum (ER)-stress-inducible drugs or protease inhibitors. This suggests that irp94 gene expression is strongly associated with the unfolded protein response (UPR) in neurons. PMID- 17913079 TI - Iridoid glucosides with insecticidal activity from Galium melanantherum. AB - The insecticidal activity of the endemic species Galium melanantherum was evaluated against Crematogaster scutellaris ants and Kalotermes flavicollis termites. Iridoid glucosides 1-7 were isolated for the first time as metabolites of the investigated plant, along with the coumarin scopolin. The main components of the extract were found to be the non-acetylated iridoids: geniposidic acid (1), 10-hydroxyloganin (2), deacetyldaphylloside (3), monotropein (4), deacetylasperulosidic acid (5) and scandoside (6), while asperulosidic acid (7) was present only in minute quantities. All isolated metabolites were identified on the basis of their spectral data. Laboratory bioassays revealed significant levels of toxicity for 1-4 against Kalotermes flavicollis termites and Crematogaster scutellaris ants. PMID- 17913080 TI - A new oviposition deterrent to the leafminer, Liriomyza trifolii: cucurbitane glucoside from Momordica charantia. AB - A new cucurbitane glucoside, 23-O-beta-D-glucopyranosyl-7-hydroxy-3-O malonylcucurbita-5,24-dien-19-al, named momordicine V, has been isolated from Momordica charantia leaves, along with the previously reported compounds, momordicines I, II, IV and 3-O-malonylmomordicine I. The structure of the new compound was established on the basis of spectral analysis, as well as by its conversion to momordicine II by alkaline catalyzed hydrolysis. Momordicine V deterred significantly the oviposition by L. trifolii on host plant leaves treated at 26.16 microg/cm2 leaf surface. PMID- 17913081 TI - Origin of renal proximal tubular injuries by Fe(III)-nta chelate. AB - Interaction between apo-transferrin and several iron(III) chelates has been investigated in terms of the capillary electrophoresis method. Based on the results, it has been clarified that (i) a binuclear iron(III) unit with an oxo bridge is necessary for the facile transfer of an iron atom from the iron(III) chelate to apo-transferrin, and (ii) the renal proximal tubular injuries by Fe(III)-nitrilotriacetate (Fe-nta) should be due to the unique binuclear structure of this complex, which gives a peroxide adduct of the binuclear Fe-nta in the presence of glutathione cycle and oxygen. PMID- 17913082 TI - Development of a new immunosensor for the detection of dopamine. AB - Graphite immunoelectrodes as immunosensors using indirect immobilization of a hapten were investigated for their applicability to detect dopamine hydrochloride at low levels. Conditions were optimized to achieve the highest sensitivity using the indirect immobilization of dopamine hydrochloride through a polymerized glutaraldehyde network on microtiter plates using ELISA technique. The conditions were later transferred to the graphite rods (phi 0.8 m x 20 mm) and a comparison between the two different sensitivities (IC50 midpoint of test) was carried out. Graphite electrodes showed higher sensitivity towards dopamine than ELISA, since they were able to detect dopamine with a midpoint of test of 0.2 mmol/l while using ELISA they were able to detect dopamine hydrochloride at 2 mmol/l. PMID- 17913083 TI - Antimicrobial activity of extracts of the lichen Parmelia sulcata and its salazinic acid constituent. AB - The antimicrobial activity of the acetone, chloroform, diethyl ether, methanol, and petroleum ether extracts of the lichen Parmelia sulcata and its salazinic acid constituent have been screened against twenty eight food-borne bacteria and fungi. All of the extracts with the exception of the petroleum ether extract showed antimicrobial activity against Aeromonas hydrophila, Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Proteus vulgaris, Yersinia enterocolitica, Staphylococcus aureus, Streptococcus faecalis, Candida albicans, Candida glabrata, Aspergillus niger, Aspergillus fumigatus, and Penicillium notatum. Salazinic acid did not show antimicrobial activity against L. monocytogenes, P. vulgaris, Y. enterocolitica, and S. faecalis but showed activity against Pseudomonas aeruginosa and Salmonella typhimurium as well. The MIC values of the extracts and the acid for the bacteria and fungi have also been determined. PMID- 17913084 TI - Mimeomal. PMID- 17913085 TI - Do I report an impaired driver? PMID- 17913086 TI - Physicians must unite to change current reimbursement situation. PMID- 17913087 TI - Celebrating women in medicine. PMID- 17913088 TI - The woman physician: drive & determination. PMID- 17913089 TI - Woman in medicine: making it work. PMID- 17913091 TI - PHP: rescuing the wounded healer--pt. I. PMID- 17913090 TI - Persistent headache: danger! PMID- 17913092 TI - 2007 workers' comp survey shows significant reduction in ortho, neurosurgery payments. PMID- 17913093 TI - Supply cost reduction in the physician's practice--part III. PMID- 17913094 TI - Ascaris lumbricoides: a travel joke? AB - Intestinal nematodes affect more than a billion people worldwide. They are commonly found in regions with poor fecal sanitation, such as developing countries. Although most of the nematode infections are non-fatal diseases, they contribute to significant morbidities such as loss of work capacity and malnutrition. We are presenting an 80-year-old male who was diagnosed with Ascaris Lumbricoides after a return from recent travel to Greece, with some clinical endoscopic images. PMID- 17913095 TI - Electrocardiographic abnormalities and endocrine diseases--part b: hypothyroidism and other thyroid diseases. PMID- 17913096 TI - The school as drug mart. PMID- 17913097 TI - Emergency Preparedness: a quick guide for school staff. PMID- 17913098 TI - Soy and corn allergies--rising in incidence and linked with other allergies. PMID- 17913099 TI - The need to prevent nicotine addiction and diabetes in our youth. The role of school health programs. PMID- 17913100 TI - Oral health education: innovative approaches across the nation help safeguard students' teeth. PMID- 17913101 TI - Nitty gritty numbers: Career and salary survey 2007. PMID- 17913103 TI - Healing magic words. PMID- 17913102 TI - Asthma care at its peak. PMID- 17913104 TI - Health + fitness-based curriculum for schools: are we ready? PMID- 17913105 TI - Making an "A" in playground injury prevention. PMID- 17913106 TI - The North Star of NMA stewardship. PMID- 17913107 TI - Why aren't there more African-American physicians? A qualitative study and exploratory inquiry of African-American students' perspectives on careers in medicine. AB - PURPOSE: African Americans comprise 13% of Americans but only 4% of U.S. physicians. The reasons for this disparity are unclear. The purpose of this study was to identify African-American high-school student perspectives on barriers to African Americans pursuing careers in medicine. METHOD: Focus group interviews (consisting of 15 questions) were conducted of African-American high-school juniors attending a Milwaukee public high school in which 89% of students are African Americans. The two focus groups were conducted in 2006, transcribed and analyzed using grounded theory. RESULTS: The 12 students interviewed in two focus groups had a mean age of 17 years; 41% of students' parents were high-school graduates. Major barriers to becoming a physician cited by students included financial constraints, lack of knowledge about medicine, little/no encouragement at home or in school, negative peer views on excelling academically, lack of African-American role models in the community and on TV, racism in medicine, and easier and more appealing alternatives for making money. Students stated that increasing the number of African-American physicians would enhance patient physician communication and relationships, and more African Americans would become physicians if there were greater exposure to medicine in schools, more guidance at a younger age and more role models. CONCLUSION: Financial constraints, insufficient exposure to medicine as a career, little encouragement at home and in schools, lack of role models, and negative peer pressure may contribute to racial disparities in the physician workforce for African Americans. Exposure at a young age to role models and to medicine as a profession might increase the number of African American physicians. PMID- 17913108 TI - "Rocking the match": applying and getting into residency. AB - Success in life requires talent and skill. But in addition, preparation is critical because it allows a person to use talent and skill to their maximal advantage, putting the person in the best position possible for whatever is planned. Success in the residency application process is more likely if the student keeps this in mind. If a student begins to prepare long before the deadlines and due dates, the process will be easier and things will be more likely to work to his or her advantage. This article is written from the perspective of someone who has evaluated many residency applications and many residency applicants. It is "an insider's view" into the process and provides some tips to medical students as they contemplate the next step after medical school. The points in the article can be remembered with the mnemonic "MRS. POPE," which should allow the student to recall the importance of Mentoring; how to prepare a Resume; the value of good Scores; the skills involved in Promoting oneself; the role of careful Observation during the interview process; and how to Present oneself and Express interest during and after the visit. Even if the mnemonic cannot be recalled, the student should remember above all that the process of preparing oneself for applying for residency begins the first day of medical school, if not sooner. Hopefully, students can use the information in this manuscript as a recipe for "rocking the match." PMID- 17913109 TI - Recruitment of underrepresented minority students to medical school: minority medical student organizations, an untapped resource. AB - Recruitment of more underrepresented minority students (black, Hispanic and native American) to increase racial diversity in the physician workforce is on the agenda for medical schools around the nation. The benefits of having a racially diverse class are indisputable. Minority physicians are more likely to provide care to minority, underserved, disadvantaged and low-income populations. Therefore, medical schools would benefit from diversity through utilizing strategies for recruitment of underrepresented minority (URM) students. Numerous recruitment strategies have been employed to increase the number of underrepresented minority students. However, formal collaboration with minority medical student organizations is an underutilized tool in the recruitment process. Many medical schools have informally used minority medical students and members of various minority organizations on campus in the recruitment process, but a formal collaboration which entails a strategic approach on using minority medical student organizations has yet to be included in the literature. This paper discusses the innovative collaboration between the University of Toledo College of Medicine (UTCOM) chapter of the Student National Medical Association (SNMA) and the college of medicine's admissions office to strategize a recruitment plan to increase the number of underrepresented minority students at the UTCOM. This paper suggests that minority medical student organizations, particularly the SNMA, can be used as a recruiting tool; hence, admissions offices cannot negate the usefulness of having formal involvement of minority medical student organizations as a recruiting tool. This approach may also be applicable to residency programs and other graduate professional fields with a severe shortage of URM students. PMID- 17913110 TI - Colorectal cancer screening of African Americans by internal medicine resident physicians can be improved with focused educational efforts. AB - Colorectal cancer causes significant morbidity and mortality in the United States. African Americans are disproportionately affected by this malignancy. There is evidence to suggest that resident physicians inconsistently screen for colorectal cancer in African Americans, perhaps because of a deficiency in knowledge and limited resources. This study evaluated internal medicine resident physicians' colorectal screening practices in African Americans prior to and following a focused educational intervention. A medical record review of internal medicine resident physicians' adherence to colorectal cancer screening recommendations was conducted. Physicians' performance of rectal exams, fecal occult blood testing, flexible sigmoidoscopy and colonoscopy was evaluated for six months prior to and six months following an educational intervention that focused upon issues related to racial disparities in colorectal cancer. Statistical significance was assessed using Fischer's exact test. There were 116 patients included in the preintervention assessment and 132 patients included in the postintervention assessment. There was no statistical significance in the rate at which rectal exams (p=0.6605) and fecal occult blood testing (p=0.7748) were performed prior to and following the educational initiative. However, there was a statistically significant difference in the rate at which endoscopic assessments (p<0.0001) were performed. Educational interventions that are focused upon racial disparity in colorectal cancer may improve resident physicians' performance of endoscopic exams in African Americans. Continued effort to enhance resident physicians' colorectal cancer screening practices in African Americans is important. PMID- 17913111 TI - Measurement of socioeconomic status in health disparities research. AB - Socioeconomic status (SES) is frequently implicated as a contributor to the disparate health observed among racial/ ethnic minorities, women and elderly populations. Findings from studies that examine the role of SES and health disparities, however, have provided inconsistent results. This is due in part to the: 1) lack of precision and reliability of measures; 2) difficulty with the collection of individual SES data; 3) the dynamic nature of SES over a lifetime; 4) the classification of women, children, retired and unemployed persons; 5) lack of or poor correlation between individual SES measures; and 6) and inaccurate or misleading interpretation of study results. Choosing the best variable or approach for measuring SES is dependent in part on its relevance to the population and outcomes under study. Many of the commonly used compositional and contextual SES measures are limited in terms of their usefulness for examining the effect of SES on outcomes in analyses of data that include population subgroups known to experience health disparities. This article describes SES measures, strengths and limitations of specific approaches and methodological issues related to the analysis and interpretation of studies that examine SES and health disparities. PMID- 17913112 TI - Use of calcaneal ultrasound and biochemical markers to assess the density and metabolic state of the bones of adults with hepatic cirrhosis. AB - Bone loss has been shown to be associated with chronic liver disease (CLD) caused by ethanol consumption or viral infection, and trabecular bone is affected more than cortical bone. We therefore used calcaneal ultrasound to compare the bone status of 54 males and 20 females with CLD in northern Nigeria with 88 age- and gender-matched healthy controls. Serum levels of bone-specific alkaline phosphatase (BSAP) and the N-terminal telopeptide of type-1 collagen (NTx) were also measured to estimate relative rates of bone synthesis and turnover, respectively. The mean stiffness index (SI) of the males with CLD and the male controls were not different; however, the mean SI of the female subjects with CLD was lower than for the female controls (101 vs. 86, p=0.003). The levels of NTx and BSAP were markedly elevated in the males, but not in the females, with CLD. Liver function tests did not correlate with ultrasound parameters or biochemical markers of bone metabolism. These results show that Nigerian women, but not males, with CLD have decreased bone density as assessed by calcaneal ultrasound; however, the high rate of bone turnover in Nigerian males with CLD indicates that they are at risk for bone loss. PMID- 17913113 TI - Hospital characteristics and racial disparities in hospital mortality from common medical conditions. AB - OBJECTIVES: Less is known about racial disparities in mortality from medical conditions than for procedures. We determined whether black-white disparities in risk-adjusted hospital mortality exist for five common conditions (myocardial infarction, congestive heart failure, cerebral vascular accident, gastrointestinal hemorrhage and pneumonia), and to determine the role of hospital characteristics. METHODS: We used the 2003 Nationwide Inpatient Sample. Where a mortality disadvantage for black patients was demonstrated, additional analyses assessed whether the degree of disparity varied by hospital characteristics. RESULTS: Mortality for black patients was equivalent to or lower than that for white patients for four of the five conditions. Black patients were more likely than white patients to die from gastrointestinal hemorrhage (1.5% vs. 1.1%, p<0.001). In multivariate analysis, hospital racial composition was the only characteristic associated with degree of disparity for gastrointestinal hemorrhage, with hospitals discharging fewer black patients demonstrating greater disparity. CONCLUSIONS: In a large, multistate sample, there was no evidence of disparities in mortality for four of five common conditions. Black-white racial disparities in mortality from gastrointestinal hemorrhage, however, may be associated with hospital racial composition. PMID- 17913114 TI - Insurance reimbursement in a university-based pediatric weight management clinic. AB - OBJECTIVES: To compare third-party payor reimbursement for patients evaluated in a university-based pediatric weight management clinic in central Kentucky. STUDY DESIGN: Demographic and reimbursement data were reviewed for 120 patients evaluated January to December 2004. Statistical analysis included Kruskal-Wallis test and Friedman's test. RESULTS: Overall, median reimbursement was 60%. For new appointments, contracted (56%) and capitated (60%) reimbursements were higher than Medicaid (55%). For established appointments, Medicaid reimbursement (100%) was higher than contracted (37%) and capitated (58%). CONCLUSION: Our data suggest that reimbursement is influenced by regional factors and is improving in central Kentucky. PMID- 17913115 TI - Prognostic factors in typhoid ileal perforation: a prospective study of 53 cases. AB - BACKGROUND: Typhoid ileal perforation remains a serious complication of typhoid enteritis with high morbidity and mortality in many tropical countries. AIMS AND OBJECTIVES: To determine the prognostic factors in typhoid perforation in Kano, Nigeria. METHODOLOGY: Fifty-three consecutive patients with typhoid perforation managed surgically were prospectively studied at the general surgical unit of Aminu Kano Teaching Hospital Kano, from March 2004 to February 2006. RESULTS: There were 26 (49.1%) males and 27 (50.9%) females, with age range of 2-55 years and a mean +/- SD of 12.2 +/- 10.2 years. The morbidity was 49.1% and the most common postoperative complications included wound infection, wound dehiscence, burst abdomen, residual intra-abdominal abscesses and enterocutaneous fistula. Mortality was 15.1% and was significantly affected by multiple perforations, severe peritoneal contamination and burst abdomen (p value <0.05, odds ratio >1). The mean duration of hospital stay for survivors was 16.1 days with a range of 8 57 days. CONCLUSION: This study has attempted to determine the factors that statistically influence mortality in typhoid perforation in our environment. PMID- 17913117 TI - Willingness of minorities to participate in biomedical studies: confirmatory findings from a follow-up study using the Tuskegee Legacy Project Questionnaire. AB - OBJECTIVES: The purposes of this analysis were to compare the self-reported willingness of blacks, Puerto-Rican Hispanics and whites to participate as research subjects in biomedical studies, and to determine the reliability of the Tuskegee Legacy Project Questionnaire (TLP). METHODS: The TLP Questionnaire, initially used in a four-city study in 1999-2000, was administered in a follow-up study within a random-digit-dial telephone survey to a stratified random sample of adults in three different U.S. cities: Baltimore, MD; New York City; and San Juan, PR. The questionnaire, a 60-item instrument, contains two validated scales: the Likelihood of Participation (LOP) Scale and the Guinea Pig Fear Factor (GPFF) Scale. RESULTS: Adjusting for age, sex, education, income and city, the LOP Scale was not statistically significantly different for the racial/ethnic groups (ANCOVA, p=87). The GPFF Scale was statistically significantly higher for blacks and Hispanics as compared to whites (adjusted ANCOVA, p<0.001). CONCLUSIONS: The of the findings from the current three-city study, as well as from our prior four city study, are remarkably similar and reinforce the conclusion that blacks and Hispanics self-report that, despite having a higher fear of participation, they are just as likely as whites to participate in biomedical research. PMID- 17913116 TI - Differences of self-reported osteoarthritis disability and race. AB - BACKGROUND: Differences in self-reported disability may be found for older black and white adults with knee osteoarthritis (OA). METHODS: This secondary analysis of data from a randomized single-blind clinical trial examined race differences in the relationship between self-reports and timed performance tests of walking. Study participants were 518 older adults (131 blacks, 387 whites), including 363 women and 155 men, with an average age of 68.6 years. RESULTS: Older black and white adults with radiographically documented knee OA reported equivalent functional ability and pain severity. However, both blacks' OA severity rating and tested performance were significantly worse than those of whites. Self-report and tested walking performance were significantly less correlated among black older adults than among white older adults. Analyses of potential confounding variables documented that the difference was not due to marital status, gender, education, income, body mass index, comorbidity, pain level, OA severity or general health. CONCLUSIONS: Self-reports of OA disability were less related to tested performance for walking among black older adults. Clinicians' knowledge of black patients' underestimation of their disability has compelling potential for improving clinical treatment and enhancing diagnostic approaches to care of older adults. PMID- 17913118 TI - The association of how time is spent during outpatient visits and patient satisfaction: are there racial differences? AB - Both satisfaction with the physician and how time is spent in the patient physician outpatient visit have been shown to differ between African-American and Caucasian patients. This study uses structural equation modeling to examine racial differences in the association between time use during the outpatient visit and patient satisfaction. This cross-sectional study employed direct observation of outpatient visits and surveys of 2,502 adult African-American and Caucasian outpatients visiting 138 primary care physicians in 84 family practices in Northeast Ohio. Patient satisfaction was measured using the Medical Outcome Study (MOS) nine-item Visit Rating Scale. Time use was assessed with the Davis Observation Code, which was used to classify every 20 seconds of a visit into 20 behavioral categories. No difference was found between African-American and Caucasian patients in the association between patient satisfaction with a physician and the time the physician spent chatting, planning treatment, providing health education, structuring the interaction, assessing health knowledge or answering patient questions. Patients were generally satisfied with their physicians, and no racial differences between Caucasians and African Americans were observed. Despite racial differences in how physicians spend time in the outpatient visit encounter, these differences are not associated with racial differences in patient satisfaction. Efforts to understand disparities in satisfaction should address areas other than how physicians allocate time in the physician-patient encounter. PMID- 17913119 TI - Cefuroxime-induced lupus. AB - Drug-induced lupus erythematosus (DILE) is a syndrome that shares symptoms and laboratory characteristics with idiopathic systemic lupus erythematosus. Recognition of DILE is important because it usually reverts within a few weeks after stopping the offending drug. Antibiotics are uncommonly associated with DILE, and cefuroxime has never been incriminated as a cause. We present herein the first case of DILE induced by cefuroxime. Although this is the first report of cefuroxime-induced DILE, we should be aware of this occurrence. PMID- 17913120 TI - Pneumomediastinum and retroperitoneal air after removal of papillomas with the microdebrider and jet ventilation. AB - OBJECTIVE: To discuss the complication of pneumothorax from alveolar rupture after transtracheal high-frequency jet ventilation and to present a case of pneumothorax, pneumomediastinum and pneumoperitoneum after jet ventilation coupled with use of the microdebrider. METHOD: Detailed case report. RESULTS: Unilateral pnuemothorax, subcutaneous emphysema, pneumomediastinum and retroperitoneal air discovered after jet ventilation for removal of airway papillomas resolved with conservative management. DISCUSSION: We discuss the difference between the respective patterns of air seepage in a peripheral alveolar injury versus a probable microperforation in the trachea. We also review the epidemiology of this rare disorder and its incidence in the African-American community. CONCLUSION: The recurrent nature of this disorder mandates multiple surgical procedures. Great care must be taken to eradicate disease and avoid complications. Pneumomediastinum in this setting can be managed conservatively. PMID- 17913121 TI - The Montreal Neurological Institute: training of the first African-American neurosurgeons. AB - BACKGROUND: Since its inception in 1934 by the legendary Dr. Wilder Penfield, the Montreal Neurological Institute (MNI) has provided world-renowned instruction in neurosurgery and related neurosciences, training many of the most prominent figures in the history of neurosurgery. Less well known is the role of the MNI in training the first African-American board-certified neurosurgeons. METHODS: A comprehensive review of pertinent modern and historical records spanning the past century was performed. RESULTS: From 1947-1965, the MNI trained the first African American board-certified neurosurgeon, and three of the first four. The first, Dr. Clarence Greene, Sr., trained at MNI from 1947-1949. The next, Dr. Jesse Barber, Jr., trained at MNI from 1958-1961. Like Greene, Barber received his MD from the Howard University College of Medicine, was on the general surgery faculty at Howard before training at MNI under Penfield and returned to Howard following his training. The third, Dr. Lloyd Dayes, matriculated at MNI in 1960 after receiving his MD from the Loma Linda University School of Medicine and trained from 1961-1965 under Dr. Theodore Rasmussen, after which he returned to Loma Linda. Greene, Barber and Dayes were certified by the American Board of Neurological Surgery in 1953, 1963 and 1967, respectively, as the first, third and fourth African-American neurosurgeons. CONCLUSION: The willingness of the world-renowned MNI to train the first African-American neurosurgeons during a time of intense racial segregation in the United States played a major role in enabling subsequent African Americans to enter and enhance the field of neurosurgery. PMID- 17913122 TI - Hydroelectricity production and forest conservation in watersheds. AB - Globally, particularly in developing countries, hydroelectricity production and economic growth occur together with ecosystem/biodiversity conservation in watersheds. There is a relationship between hydroelectricity production and ecosystem/biodiversity conservation in watersheds, centering on the supply and demand for ecosystem services of river water flow regulation and sediment retention. Here we show that, in the upper reach of the Yangtze River, hydroelectricity production of Three Gorges Hydroelectric Power Plant can form a beneficial relationship with forest conservation through the paid use (compensating residents for their cooperation in the conservation) of ecosystem services launched by the National Natural Forest Protection Project. This interaction can provide additional incentives to encourage local communities' long-term cooperation in conserving and protecting the restored forest ecosystems. Hydroelectricity plants also obtain benefits from this interaction. The industrialization of ecosystem services supply provides an operational framework for this beneficial interaction. Sustainable forest ecosystem conservation will require developing new institutions and policies and must involve local communities in the conservation and protection of their local forests. PMID- 17913123 TI - Willow on Yellowstone's northern range: evidence for a trophic cascade? AB - Reintroduction of wolves (Canis lupus) to Yellowstone National Park in 1995-1996 has been argued to promote a trophic cascade by altering elk (Cervus elaphus) density, habitat-selection patterns, and behavior that, in turn, could lead to changes within the plant communities used by elk. We sampled two species of willow (Salix boothii and S. geyeriana) on the northern winter range to determine whether (1) there was quantitative evidence of increased willow growth following wolf reintroduction, (2) browsing by elk affected willow growth, and (3) any increase in growth observed was greater than that expected by climatic and hydrological factors alone, thereby indicating a trophic cascade caused by wolves. Using stem sectioning techniques to quantify historical growth patterns we found an approximately twofold increase in stem growth-ring area following wolf reintroduction for both species of willow. This increase could not be explained by climate and hydrological factors alone; the presence of wolves on the landscape was a significant predictor of stem growth above and beyond these abiotic factors. Growth-ring area was positively correlated with the previous year's ring area and negatively correlated with the percentage of twigs browsed from the stem during the winter preceding growth, indicating that elk browse impeded stem growth. Our results are consistent with the hypothesis of a behaviorally mediated trophic cascade on Yellowstone's northern winter range following wolf reintroduction. We suggest that the community-altering effects of wolf restoration are an endorsement of ecological-process management in Yellowstone National Park. PMID- 17913124 TI - Hydrologic regime and herbivory stabilize an alternative state in Yellowstone National Park. AB - A decline in the stature and abundance of willows during the 20th century occurred throughout the northern range of Yellowstone National Park, where riparian woody-plant communities are key components in multiple-trophic-level interactions. The potential causes of willow decline include climate change, increased elk browsing coincident with the loss of an apex predator, the gray wolf, and an absence of habitat engineering by beavers. The goal of this study was to determine the spatial and temporal patterns of willow establishment through the 20th century and to identify causal processes. Sampled willows established from 1917 to 1999 and contained far fewer young individuals than was predicted from a modeled stable willow population, indicating reduced establishment during recent decades. Two hydrologically distinct willow establishment environments were identified: fine-grained beaver pond sediments and coarse-grained alluvium. Willows established on beaver pond sediment earlier in time, higher on floodplain surfaces, and farther from the current stream channel than did willows on alluvial sediment. Significant linear declines from the 1940s to the 1990s in alluvial willow establishment elevation and lateral distance from the stream channel resulted in a much reduced area of alluvial willow establishment. Willow establishment was not well correlated with climate driven hydrologic variables, but the trends were consistent with the effects of stream channel incision initiated in ca. 1950, 20-30 years after beaver dam abandonment. Radiocarbon dates and floodplain stratigraphy indicate that stream incision of the present magnitude may be unprecedented in the past two millennia. We propose that hydrologic changes, stemming from competitive exclusion of beaver by elk overbrowsing, caused the landscape to transition from a historical beaver pond and willow-mosaic state to its current alternative stable state where active beaver dams and many willow stands are absent. Because of hydrologic changes in streams, a rapid return to the historical state may not occur by reduction of elk browsing alone. Management intervention to restore the historical hydrologic regime may be necessary to recover willows and beavers across the landscape. PMID- 17913125 TI - Evaluating prey switching in wolf-ungulate systems. AB - Wolf restoration has become a widely accepted conservation and management practice throughout North America and Europe, though the ecosystem effects of returning top carnivores remain both scientific and societal controversies. Mathematical models predicting and describing wolf-ungulate interactions are typically limited to the wolves' primary prey, with the potential for prey switching in wolf-multiple-ungulate systems only suggested or assumed by a number of investigators. We used insights gained from experiments on small taxa and field data from ongoing wolf-ungulate studies to construct a model of predator diet composition for a wolf-elk-bison system in Yellowstone National Park, Wyoming, USA. The model explicitly incorporates differential vulnerability of the ungulate prey types to predation, predator preference, differences in prey biomass, and the possibility of prey switching. Our model demonstrates wolf diet shifts with changes in relative abundance of the two prey, with the dynamics of this shift dependent on the combined influences of preference, differential vulnerability, relative abundances of prey, and whether or not switching occurs. Differences in vulnerability between elk and bison, and strong wolf preference for elk, result in an abrupt dietary shift occurring only when elk are very rare relative to bison, whereas incorporating switching initiates the dietary shift more gradually and at higher bison-elk ratios. We demonstrate how researchers can apply these equations in newly restored wolf-two-prey systems to empirically evaluate whether prey switching is occurring. Each coefficient in the model has a biological interpretation, and most can be directly estimated from empirical data collected from field studies. Given the potential for switching to dramatically influence predator-prey dynamics and the wide range of expected prey types and abundances in some systems where wolves are present and/or being restored, we suggest that this is an important and productive line of research that should be pursued by ecologists working in wolf-ungulate systems. PMID- 17913126 TI - Irruptive population dynamics in Yellowstone pronghorn. AB - Irruptive population dynamics appear to be widespread in large herbivore populations, but there are few empirical examples from long time series with small measurement error and minimal harvests. We analyzed an 89-year time series of counts and known removals for pronghorn (Antilocapra americana) in Yellowstone National Park of the western United States during 1918-2006 using a suite of density-dependent, density-independent, and irruptive models to determine if the population exhibited irruptive dynamics. Information-theoretic model comparison techniques strongly supported irruptive population dynamics (Leopold model) and density dependence during 1918-1946, with the growth rate slowing after counts exceeded 600 animals. Concerns about sagebrush (Artemisia spp.) degradation led to removals of >1100 pronghorn during 1947-1966, and counts decreased from approximately 700 to 150. The best models for this period (Gompertz, Ricker) suggested that culls replaced intrinsic density-dependent mechanisms. Contrary to expectations, the population did not exhibit enhanced demographic vigor soon after the termination of the harvest program, with counts remaining between 100 and 190 animals during 1967 1981. However, the population irrupted (Caughley model with a one-year lag) to a peak abundance of approximately 600 pronghorn during 1982-1991, with a slowing in growth rate as counts exceeded 500. Numbers crashed to 235 pronghorn during 1992-1995, perhaps because important food resources (e.g., sagebrush) on the winter range were severely diminished by high densities of browsing elk, mule deer, and pronghorn. Pronghorn numbers remained relatively constant during 1996-2006, at a level (196-235) lower than peak abundance, but higher than numbers following the release from culling. The dynamics of this population supported the paradigm that irruption is a fundamental pattern of growth in many populations of large herbivores with high fecundity and delayed density-dependent effects on recruitment when forage and weather conditions become favorable after range expansion or release from harvesting. Incorporating known removals into population models that can describe a wide range of dynamics can greatly improve our interpretation of observed dynamics in intensively managed populations. PMID- 17913127 TI - Selective harvesting and habitat loss produce long-term life history changes in a mouflon population. AB - We examined the long-term effects (28 years) of habitat loss and phenotype-based selective harvest on body mass, horn size, and horn shape of mouflon (Ovis gmelini musimon) in southern France. This population has experienced habitat deterioration (loss of 50.8% of open area) since its introduction in 1956 and unrestricted selective hunting of the largest horned males since 1973. Both processes are predicted to lead to a decrease in phenotype quality by decreasing habitat quality and by reducing the reproductive contribution of individuals carrying traits that are targeted by hunters. Body mass and body size of both sexes and horn measurements of males markedly decreased (by 3.4-38.3%) in all age classes from the 1970s. Lamb body mass varied in relation to the spatiotemporal variation of habitat closure within the hunting-free reserve, suggesting that habitat closure explains part of these changes. However, the fact that there was no significant spatial variation in body mass in the early part of the study, when a decline in phenotypic quality already had occurred, provided support for the influence of selective harvesting. We also found that the allometric relationship between horn breadth and horn length changed over the study period. For a given horn length, horn breadth was lower during the second part of the study. This result, as well as changes in horn curve diameter, supports the interpretation that selective harvesting of males based on their horn configuration had evolutionary consequences for horn shape, since this phenotypic trait is less likely to be affected by changes in habitat characteristics. Moreover, males required more time (approximately four years) to develop a desirable trophy, suggesting that trophy hunting favors the reproductive contribution of animals with slow-growing horns. Managers should exploit hunters' desire for trophy males to finance management strategies which ensure a balance between the population and its environment. However, for long-term sustainable exploitation, harvest strategy should also ensure that selectively targeted males are allowed to contribute genetically to the next generations. PMID- 17913128 TI - Forest productivity decline caused by successional paludification of boreal soils. AB - Long-term forest productivity decline in boreal forests has been extensively studied in the last decades, yet its causes are still unclear. Soil conditions associated with soil organic matter accumulation are thought to be responsible for site productivity decline. The objectives of this study were to determine if paludification of boreal soils resulted in reduced forest productivity, and to identify changes in the physical and chemical properties of soils associated with reduction in productivity. We used a chronosequence of 23 black spruce stands ranging in postfire age from 50 to 2350 years and calculated three different stand productivity indices, including site index. We assessed changes in forest productivity with time using two complementary approaches: (1) by comparing productivity among the chronosequence stands and (2) by comparing the productivity of successive cohorts of trees within the same stands to determine the influence of time independently of other site factors. Charcoal stratigraphy indicates that the forest stands differ in their fire history and originated either from high- or low-severity soil burns. Both chronosequence and cohort approaches demonstrate declines in black spruce productivity of 50-80% with increased paludification, particularly during the first centuries after fire. Paludification alters bryophyte abundance and succession, increases soil moisture, reduces soil temperature and nutrient availability, and alters the vertical distribution of roots. Low-severity soil burns significantly accelerate rates of paludification and productivity decline compared with high-severity fires and ultimately reduce nutrient content in black spruce needles. The two combined approaches indicate that paludification can be driven by forest succession only, independently of site factors such as position on slope. This successional paludification contrasts with edaphic paludification, where topography and drainage primarily control the extent and rate of paludification. At the landscape scale, the fire regime (frequency and severity) controls paludification and forest productivity through its effect on soil organic layers. Implications for global carbon budgets and sustainable forestry are discussed. PMID- 17913129 TI - Influences of secondary disturbances on lodgepole pine stand development in Rocky Mountain National Park. AB - Although high-severity fire is the primary type of disturbance shaping the structure of lodgepole pine (Pinus contorta) stands in the southern Rocky Mountains, many post-fire stands are also affected by blowdown, low-severity surface fires, and/or outbreaks of mountain pine beetle (MPB; Dendroctonus ponderosae). The ecological effects of these secondary disturbances are poorly understood but are potentially important in the context of managing for ecological restoration and fire hazard mitigation. We investigated the effects of blowdown, surface fires, and MPB outbreaks on demographic processes in post-fire lodgepole pine stands in Rocky Mountain National Park, Colorado, USA. We used dendroecological methods to reconstruct stand characteristics prior to and following secondary disturbances for paired stands with and without secondary disturbances. Surface fire events do not kill canopy trees or trigger pulses of recruitment and as such do not have detectable influences on stand development. In contrast, both MPB and blowdown kill canopy trees and trigger pulses of tree regeneration of lodgepole pine and subalpine fir (Abies lasiocarpa). The amount and species composition of post-disturbance regeneration is dependent on the severity of the disturbance and on the time since stand initiation. Secondary disturbances of higher severity (i.e., killing >50% of the canopy trees) that occur in younger post-fire stands favor new establishment of lodgepole pine. In contrast, secondary disturbances of lower severity in older stands (>250 years) trigger a pulse of establishment of subalpine fir. The results of this study demonstrate that the high tree densities characteristic of lodgepole pine stands in the southern Rockies (southern Wyoming to northern New Mexico) are the result of dense regeneration following stand-replacing fires and that surface fires had little or no thinning effect on tree densities. Thus, current high stand densities in the study area are not the result of suppression of surface fires. Moreover, the strong pulses of regeneration following forest thinning by MPB and blowdowns imply that, depending on the degree of thinning, thinning prescriptions to reduce fuels in the lodgepole pine forest type may have the unintended consequence of increasing ladder fuels 15-20 years following treatments. PMID- 17913130 TI - Rapid assessment of postfire plant invasions in coniferous forests of the western United States. AB - Fire is a natural part of most forest ecosystems in the western United States, but its effects on nonnative plant invasion have only recently been studied. Also, forest managers are engaging in fuel reduction projects to lessen fire severity, often without considering potential negative ecological consequences such as nonnative plant species introductions. Increased availability of light, nutrients, and bare ground have all been associated with high-severity fires and fuel treatments and are known to aid in the establishment of nonnative plant species. We use vegetation and environmental data collected after wildfires at seven sites in coniferous forests in the western United States to study responses of nonnative plants to wildfire. We compared burned vs. unburned plots and plots treated with mechanical thinning and/or prescribed burning vs. untreated plots for nonnative plant species richness and cover and used correlation analyses to infer the effect of abiotic site conditions on invasibility. Wildfire was responsible for significant increases in nonnative species richness and cover, and a significant decrease in native cover. Mechanical thinning and prescribed fire fuel treatments were associated with significant changes in plant species composition at some sites. Treatment effects across sites were minimal and inconclusive due to significant site and site x treatment interaction effects caused by variation between sites including differences in treatment and fire severities and initial conditions (e.g., nonnative species sources). We used canonical correspondence analysis (CCA) to determine what combinations of environmental variables best explained patterns of nonnative plant species richness and cover. Variables related to fire severity, soil nutrients, and elevation explained most of the variation in species composition. Nonnative species were generally associated with sites with higher fire severity, elevation, percentage of bare ground, and lower soil nutrient levels and lower canopy cover. Early assessments of postfire stand conditions can guide rapid responses to nonnative plant invasions. PMID- 17913131 TI - Regional extent of an ecosystem engineer: earthworm invasion in northern hardwood forests. AB - The invasion of exotic earthworms into northern temperate and boreal forests previously devoid of earthworms is an important driver of ecosystem change. Earthworm invasion can cause significant changes in soil structure and communities, nutrient cycles, and the diversity and abundance of herbaceous plants. However, the regional extent and patterns of this invasion are poorly known. We conducted a regional survey in the Chippewa and Chequamegon National Forests, in Minnesota and Wisconsin, U.S.A., respectively, to measure the extent and patterns of earthworm invasion and their relationship to potential earthworm introduction sites. We sampled earthworms, soils, and vegetation in 20 mature, sugar maple-dominated forest stands in each national forest and analyzed the relationship between the presence of five earthworm taxonomic groups, habitat variables, and distance to the nearest potential introduction site. Earthworm invasion was extensive but incomplete in the two national forests. Four of the six earthworm taxonomic groups occurred in 55-95% of transects; however 20% of all transects were invaded by only one taxonomic group that has relatively minor ecological effects. Earthworm taxonomic groups exhibited a similar sequence of invasion found in other studies: Dendrobaena > Aporrectodea = Lumbricus juveniles > L. rubellus > L. terrestris. Distance to the nearest road was the best predictor of earthworm invasion in Wisconsin while distance to the nearest cabin was the best predictor in Minnesota. These data allow us to make preliminary assessments of landscape patterns of earthworm invasion. As an example, we estimate that 82% of upland mesic hardwood stands in the Wisconsin region are likely invaded by most taxonomic groups while only 3% are unlikely to be invaded at present. Distance to roads and cabins provides a coarse-scale predictor of earthworm invasion to focus stand-level assessments that will help forest managers better understand current and potential forest conditions and identify uninvaded areas that could serve as important refugia for plant species threatened by earthworm invasion. PMID- 17913132 TI - Mangrove recruitment after forest disturbance is facilitated by herbaceous species in the Caribbean. AB - Plant communities along tropical coastlines are often affected by natural and human disturbances, but little is known about factors influencing recovery. We focused on mangrove forests, which are among the most threatened ecosystems globally, to examine how facilitation by herbaceous vegetation might improve forest restoration after disturbance. We specifically investigated whether recovery of mangrove forests in harsh environments is accelerated by nurse plants and whether the beneficial effects are species-specific. Quantification of standardized effects allowed comparisons across performance parameters and over time for: (1) net effect of each herbaceous species on mangrove survival and growth, (2) effects of pre- and post-establishment factors associated with each herbaceous species, and (3) need for artificial planting to enhance growth or survival of mangrove seedlings. Mangrove recruitment in a clear-cut forest in Belize was accelerated by the presence of Sesuvium portulacastrum (succulent forb) and Distichlis spicata (grass), two coastal species common throughout the Caribbean region. The net effect of herbaceous vegetation was positive, but the magnitude of effects on mangrove survival and growth differed by species. Because of differences in their vegetative structure and other features, species effects on mangroves also varied by mechanism: (1) trapping of dispersing propagules (both species), (2) structural support of the seedling (Distichlis), and/or (3) promotion of survival (Sesuviumn) or growth (Distichlis) through amelioration of soil conditions (temperature, aeration). Artificial planting had a stronger positive effect on mangrove survival than did edaphic conditions, but planting enhanced mangrove growth more in Sesuvium than in Distichlis patches. Our study indicates that beneficial species might be selected based on features that provide multiple positive effects and that species comparisons may be improved using standardized effects. Our findings are not only relevant to the coastal environments found in the Caribbean region, but our assessment methods may be useful for developing site-specific information to restore disturbed mangrove forests worldwide, especially given the large pool of mangrove associates (>45 genera) available for screening. PMID- 17913133 TI - Endangered light-footed clapper rail affects parasite community structure in coastal wetlands. AB - An extinction necessarily affects community members that have obligate relationships with the extinct species. Indirect or cascading effects can lead to even broader changes at the community or ecosystem level. However, it is not clear whether generalist parasites should be affected by the extinction of one of their hosts. We tested the prediction that loss of a host species could affect the structure of a generalist parasite community by investigating the role of endangered Light-footed Clapper Rails (Rallus longirostris levipes) in structuring trematode communities in four tidal wetlands in southern California, U.S.A. (Carpinteria Salt Marsh, Mugu Lagoon) and Mexico (Estero de Punta Banda, Bahia Falsa-San Quintin). We used larval trematode parasites in first intermediate host snails (Cerithidea californica) as windows into the adult trematodes that parasitize Clapper Rails. Within and among wetlands, we found positive associations between Clapper Rails and four trematode species, particularly in the vegetated marsh habitat where Clapper Rails typically occur. This suggests that further loss of Clapper Rails is likely to affect the abundance of several competitively dominant trematode species in wetlands with California horn snails, with possible indirect effects on the trematode community and changes in the impacts of these parasites on fishes and invertebrates. PMID- 17913134 TI - Effects of anthropogenic developments on common raven nesting biology in the West Mojave Desert. AB - Subsidized predators may affect prey abundance, distribution, and demography. Common Ravens (Corvus corax) are anthropogenically subsidized throughout their range and, in the Mojave Desert, have increased in number dramatically over the last 3-4 decades. Human-provided food resources are thought to be important drivers of raven population growth, but human developments add other features as well, such as nesting platforms. From 1996 to 2000, we examined the nesting ecology of ravens in the Mojave Desert, relative to anthropogenic developrhent. Ravens nested disproportionately near point sources of food and water subsidies (such as towns, landfills, and ponds) but not near roads (sources of road-killed carrion), even though both sources of subsidy enhanced fledging success. Initiation of breeding activity was more likely when a nest from the previous year was present at the start of a breeding season but was not affected by access to food. The relative effect of environmental modifications on fledging success varied from year to year, but the effect of access to human-provided resources was comparatively consistent, suggesting that humans provide consistently high quality breeding habitat for ravens. Anthropogenic land cover types in the desert are expected to promote raven population growth and to allow ravens to occupy parts of the desert that otherwise would not support them. Predatory impacts of ravens in the Mojave Desert can therefore be considered indirect effects of anthropogenic development. PMID- 17913135 TI - Habitat classification modeling with incomplete data: pushing the habitat envelope. AB - Habitat classification models (HCMs) are invaluable tools for species conservation, land-use planning, reserve design, and metapopulation assessments, particularly at broad spatial scales. However, species occurrence data are often lacking and typically limited to presence points at broad scales. This lack of absence data precludes the use of many statistical techniques for HCMs. One option is to generate pseudo-absence points so that the many available statistical modeling tools can bb used. Traditional techniques generate pseudo absence points at random across broadly defined species ranges, often failing to include biological knowledge concerning the species-habitat relationship. We incorporated biological knowledge of the species-habitat relationship into pseudo absence points by creating habitat envelopes that constrain the region from which points were randomly selected. We define a habitat envelope as an ecological representation of a species, or species feature's (e.g., nest) observed distribution (i.e., realized niche) based on a single attribute, or the spatial intersection of multiple attributes. We created HCMs for Northern Goshawk (Accipiter gentilis atricapillus) nest habitat during the breeding season across Utah forests with extant nest presence points and ecologically based pseudo absence points using logistic regression. Predictor variables were derived from 30-m USDA Landfire and 250-m Forest Inventory and Analysis (FIA) map products. These habitat-envelope-based models were then compared to null envelope models which use traditional practices for generating pseudo-absences. Models were assessed for fit and predictive capability using metrics such as kappa, threshold independent receiver operating characteristic (ROC) plots, adjusted deviance (D(adj)2), and cross-validation, and were also assessed for ecological relevance. For all cases, habitat envelope-based models outperformed null envelope models and were more ecologically relevant, suggesting that incorporating biological knowledge into pseudo-absence point generation is a powerful tool for species habitat assessments. Furthermore, given some a priori knowledge of the species habitat relationship, ecologically based pseudo-absence points can be applied to any species, ecosystem, data resolution, and spatial extent. PMID- 17913136 TI - The importance of protected areas for the forest and endemic avifauna of Sulawesi (Indonesia). AB - Protected areas are critical for the conservation of residual tropical forest biodiversity, yet many of these are being deforested by humans both within and outside of their administrative boundaries. Therefore, it is critical to document the significance of protected areas for conserving tropical biodiversity, particularly in mega-diverse Southeast Asia. We evaluated the importance of protected areas (national parks [NP], nature reserves [NR], and wildlife reserves [WR]) in preserving avifaunal diversity, particularly the endemic and forest species, on the island of Sulawesi. This island has one of the highest numbers of endemic avifauna genera (12) globally and is also experiencing heavy deforestation. Rarefaction analyses and species estimators showed that parks and reserves consistently recorded higher number of forest, endemic, and endemic forest bird species, in addition to larger population densities, than in their surrounding human-modified areas across eight protected areas (Gunung Manembo nembo WR, Tangkoko-Batu Angus and Dua Saudara NR, Gunung Ambang NR, Bogani Nani Wartabone NP, Gunung Tinombala NR, Gunung Sojol NR, Lore Lindu NP, and Rawa Aopa Watumohai NP). This implies that protecting natural forests must remain as one of the fundamental conservation strategies in Sulawesi. Two small reserves (Gunung Manembo-nembo WR and Tangkoko-Batu Angus and Dua Saudara NR), however, had high number of forest and endemic bird species both within and outside their boundaries, suggesting the importance of buffer areas for augmenting small reserves so as to improve their conservation value. Ordination analyses revealed the differential response of bird species to different environmental factors (e.g., native tree cover), highlighting the significance of forested habitats with dense native vegetation cover for effective conservation of forest dependent and endemic avifauna. In addition, the distinctiveness in bird species composition among protected areas highlights the importance of establishing a reserve network across major altitudinal zones so as to achieve maximum complementarity for the conservation of Sulawesi's unique avifauna. PMID- 17913137 TI - A cross-regional assessment of the factors affecting ecoliteracy: implications for policy and practice. AB - The value of accumulated ecological knowledge, termed ecoliteracy, is vital to both human and ecosystem health. Maintenance of this knowledge is essential for continued support of local conservation efforts and the capacity of communities to self- or co-manage their local resources sustainably. Most previous studies have been qualitative and small scale, documenting ecoliteracy in geographically isolated locations. In this study, we take a different approach, focusing on (1) the primary factors affecting individual levels of ecoliteracy, (2) whether these factors shift with economic development, and (3) if different knowledge protection strategies are required for the future. We compared non-resource dependent communities in the United Kingdom with resource-dependent communities in India and Indonesia (n=1250 interviews). We found that UK residents with the highest levels of ecoliteracy visited the countryside frequently, lived and grew up in rural areas, and acquired their knowledge from informal word-of-mouth sources, such as parents and friends, rather than television and schooling. The ecoliteracy of resource-dependent community members, however, varied with wealth status and gender. The least wealthy families depended most on local resources for their livelihoods and had the highest levels of ecoliteracy. Gender roles affected both the level and content of an individual's ecoliteracy. The importance of reciprocal oral transfer of this knowledge in addition to direct experience to the maintenance of ecoliteracy was apparent at all sites. Lessons learned may contribute to new local resource management strategies for combined ecoliteracy conservation. Without novel policies, local community management capacity is likely to be depleted in the future. PMID- 17913138 TI - Serial depletion of marine invertebrates leads to the decline of a strongly interacting grazer. AB - We investigated the relative roles of natural factors and shoreline harvest leading to recent declines of the black leather chiton (Katharina tunicata) on the outer Kenai Peninsula, Alaska (U.S.A.). This intertidal mollusk is a strongly interacting grazer and a culturally important subsistence fishery for Sugpiaq (Chugach Alutiiq) natives. We took multiple approaches to determine causes of decline. Field surveys examined the significant predictors of Katharina density and biomass across 11 sites varying in harvest pressure, and an integrated analysis of archaeological faunal remains, historical records, traditional ecological knowledge, and contemporary subsistence invertebrate landings examined changes in subsistence practices through time. Strong evidence suggests that current spatial variation in Katharina density and biomass is driven by both human exploitation and sea otter (Enhydra lutris) predation. Traditional knowledge, calibrated by subsistence harvest data, further revealed that several benthic marine invertebrates (sea urchin, crab, clams, and cockles) have declined serially beginning in the 1960s, with reduced densities and sizes of Katharina being the most recent. The timing of these declines was coincident with changes in human behavior (from semi-nomadic to increasingly permanent settlement patterns, improved extractive technologies, regional commercial crustacean exploitation, the erosion of culturally based season and size restrictions) and with the reestablishment of sea otters. We propose that a spatial concentration in shoreline collection pressure through time, increased harvest efficiency, and the serial depletion of alternative marine invertebrate prey have led to intensified per capita predator impacts on Katharina and thus its recent localized decline. PMID- 17913139 TI - Graph theory as a proxy for spatially explicit population models in conservation planning. AB - Spatially explicit population models (SEPMs) are often considered the best way to predict and manage species distributions in spatially heterogeneous landscapes. However, they are computationally intensive and require extensive knowledge of species' biology and behavior, limiting their application in many cases. An alternative to SEPMs is graph theory, which has minimal data requirements and efficient algorithms. Although only recently introduced to landscape ecology, graph theory is well suited to ecological applications concerned with connectivity or movement. This paper compares the performance of graph theory to a SEPM in selecting important habitat patches for Wood Thrush (Hylocichla mustelina) conservation. We use both models to identify habitat patches that act as population sources and persistent patches and also use graph theory to identify patches that act as stepping stones for dispersal. Correlations of patch rankings were very high between the two models. In addition, graph theory offers the ability to identify patches that are very important to habitat connectivity and thus long-term population persistence across the landscape. We show that graph theory makes very similar predictions in most cases and in other cases offers insight not available from the SEPM, and we conclude that graph theory is a suitable and possibly preferable alternative to SEPMs for species conservation in heterogeneous landscapes. PMID- 17913140 TI - Understanding uncertainty in the effect of low-head dams on fishes of Great Lakes tributaries. AB - Small dams represent one of the most widespread human influences on riverscapes. Greater understanding of how these structures affect aquatic organisms is needed to ensure that decisions regarding their construction and removal strike an appropriate balance between components of human and ecosystem services. Within the basin of the Laurentian Great Lakes, the effects that in-stream barriers (dams) used to control the non-native, parasitic sea lamprey (Petromyzon marinus) on the diversity of non-target fishes is a significant concern for fishery managers. A previous study indicated that upstream changes in the species richness of non-target fishes observed in 24 streams with a sea lamprey barrier relative to paired reference streams (a measure of effect size) was variable across the basin. We examined the degree to which the variance in effect size could be attributed to imprecision in the field sampling protocol used to estimate effect sizes, differences in catchment-scale landscape attributes between barrier and reference streams within pairs, and differences in landscape attributes at different spatial scales among barrier streams. Simulation modeling and analyses of repeated field measurements made for a subset of streams demonstrated that a large variance in effect size is expected for the field sampling design and that estimates of effect size measured for individual barrier streams are imprecise. Regression models and multimodel inference methods based on Akaike's Information Criterion provided less support for hypotheses linking effect size to landscape attributes. Mean effect size, adjusted for the influences of landscape characteristics within and across stream pairs, provides the most reliable and least biased estimate of the effect of sea lamprey barriers on the richness of nontarget fish species. With the information currently available, landscape characteristics of catchments cannot be used to help decision makers anticipate effects sizes for candidate streams being considered for future barrier construction. Our findings will help fishery managers in the Laurentian Great Lakes make more informed decisions regarding the use and placement of sea lamprey barriers and achieve their objective of delivering an integrated pest management plan for sea lamprey control that is environmentally and economically sound and socially acceptable. PMID- 17913141 TI - Stream ecosystem functioning under reduced flow conditions. AB - Assessments of flow reduction in streams often focus on changes to biological communities and in-stream physical characteristics, with little consideration for changes in ecosystem functioning. It is unclear whether functional indicators of ecosystem condition may be useful for assessing the impacts of reduced discharge on small streams. Using weirs and diversions to reduce stream discharge during summer baseflow conditions, we tested the response of leaf breakdown, coarse particulate organic matter (CPOM) retention, and primary production to one month of water abstraction in before-after, control-impact (BACI) designed experiments. Discharge at impact (downstream) reaches decreased by over 85% in each of three small New Zealand streams compared to controls (upstream). There also were decreases in velocity, depth, and wetted width. Sediment cover increased at impact reaches, but there were only small changes to conductivity, pH, and surface water temperature. We installed mesh bags filled with willow leaves in stream for one month to measure leaf breakdown. Reduced discharge had little influence on leaf breakdown rate in these streams. Travel distances and retention structures for CPOM were evaluated using releases of paper strips and wooden dowelling over a range of discharges. The distance traveled by released CPOM increased with increasing discharge, and the importance of riffles as retention structures increased at lower discharges. We measured the accumulation of chlorophyll a after one month on artificial substrates as an estimate of the relative primary production of control and impact reaches. The differences in chlorophyll a concentrations between control and impact reaches were inconsistent among streams. These ecosystem functions have responded inconsistently to water removal in these streams. However, the strong response of CPOM retention to reduced discharge could complement measures of biological community structure when the influence of reduced discharge is assessed. We recommend further investigation in a wide range of streams to assess the utility of these processes as functional indicators of reduced discharge. PMID- 17913142 TI - Interrelationships among shrub encroachment, land management, and litter decomposition in a semidesert grassland. AB - Encroachment of woody plants into grasslands, and subsequent brush management, are among the most prominent changes to occur in arid and semiarid systems over the past century. Despite the resulting widespread changes in landcover, substantial uncertainty about the biogeochemical impacts of woody proliferation and brush management exists. We explored the role of shrub encroachment and brush management on leaf litter decomposition in a semidesert grassland where velvet mesquite (Prosopis velutina) abundance has increased over the past 100 years. This change in physiognomy may affect decomposition directly, through altered litter quality or quantity, and indirectly through altered canopy structure. To assess the direct and indirect impacts of shrubs on decomposition, we quantified changes in mass, nitrogen, and carbon in litterbags deployed under mesquite canopies and in intercanopy zones. Litterbags contained foliage from mesquite and Lehmann lovegrass (Eragrostis lehmanniana), a widespread, nonnative grass in southern Arizona. To explore short- and long-term influences of brush management on the initial stages of decomposition, litterbags were deployed at sites where mesquite canopies were removed three weeks, 45 years, or 70 years prior to study initiation. Mesquite litter decomposed more rapidly than lovegrass, but negative indirect influences of mesquite canopies counteracted positive direct effects. Decomposition was positively correlated with soil infiltration into litterbags, which varied with microsite placement, and was lowest under canopies. Low under canopy decomposition was ostensibly due to decreased soil movement associated with high under-canopy herbaceous biomass. Decomposition rates where canopies were removed three weeks prior to study initiation were comparable to those beneath intact canopies, suggesting that decomposition was driven by mesquite legacy effects on herbaceous cover-soil movement linkages. Decomposition rates where shrubs were removed 45 and 70 years prior to study initiation were comparable to intercanopy rates, suggesting that legacy effects persist less than 45 years. Accurate decomposition modeling has proved challenging in arid and semiarid systems but is critical to understanding biogeochemical responses to woody encroachment and brush management. Predicting brush-management effects on decomposition will require information on shrub-grass interactions and herbaceous biomass influences on soil movement at decadal timescales. Inclusion of microsite factors controlling soil accumulation on litter would improve the predictive capability of decomposition models. PMID- 17913143 TI - Grassland invader responses to realistic changes in native species richness. AB - The importance of species richness for repelling exotic plant invasions varies from ecosystem to ecosystem. Thus, in order to prioritize conservation objectives, it is critical to identify those ecosystems where decreasing richness will most greatly magnify invasion risks. Our goal was to determine if invasion risks greatly increase in response to common reductions in grassland species richness. We imposed treatments that mimic management-induced reductions in grassland species richness (i.e., removal of shallow- and/or deep-rooted forbs and/or grasses and/or cryptogam layers). Then we introduced and monitored the performance of a notorious invasive species (i.e., Centaurea maculosa). We found that, on a per-gram-of-biomass basis, each resident plant group similarly suppressed invader growth. Hence, with respect to preventing C. maculosa invasions, maintaining overall productivity is probably more important than maintaining the productivity of particular plant groups or species. But at the sites we studied, all plant groups may be needed to maintain overall productivity because removing forbs decreased overall productivity in two of three years. Alternatively, removing forbs increased productivity in another year, and this led us to posit that removing forbs may inflate the temporal productivity variance as opposed to greatly affecting time-averaged productivity. In either case, overall productivity responses to single plant group removals were inconsistent and fairly modest, and only when all plant groups were removed did C. maculosa growth increase substantially over a no-removal treatment. As such, it seems that intense disturbances (e.g., prolonged drought, overgrazing) that deplete multiple plant groups may often be a prerequisite for C. maculosa invasion. PMID- 17913144 TI - Predicting the economic impact of an invasive species on an ecosystem service. AB - Quantifying the impact of alien invasive species on ecosystem services is an essential step in developing effective practices and policy for invasive species management. Here we develop a stochastic bioeconomic model that enables the economic impact of an invasive pest to be estimated before its arrival, based on relatively poorly specified ecological and economic parameters. We developed the model by using a hypothetical invasion of the varroa bee mite (Varroa destructor) into Australia and the negative flow-on effects that it would have on pollination by reducing honey bee populations, giving rise to a loss of pollination services, reduced crop yields, and additional production costs. If the mite were to continue to be prevented from entering the country over the next 30 years, we estimate that the economic costs avoided would be U.S. $16.4-38.8 million (Aus $21.3-50.5 million) per year. We suggest that current invasion response funding arrangements in Australia, which do not acknowledge these avoided damages, require amendment. PMID- 17913145 TI - Caveats to quantifying ecosystem services: fruit abortion blurs benefits from crop pollination. AB - The recent trend to place monetary values on ecosystem services has led to studies on the economic importance of pollinators for agricultural crops. Several recent studies indicate regional, long-term pollinator declines, and economic consequences have been derived from declining pollination efficiencies. However, use of pollinator services as economic incentives for conservation must consider environmental factors such as drought, pests, and diseases, which can also limit yields. Moreover, "flower excess" is a well-known reproductive strategy of plants as insurance against unpredictable, external factors that limit reproduction. With three case studies on the importance of pollination levels for amounts of harvested fruits of three tropical crops (passion fruit in Brazil, coffee in Ecuador, and cacao in Indonesia) we illustrate how reproductive strategies and environmental stress can obscure initial benefits from improved pollination. By interpreting these results with findings from evolutionary sciences, agronomy, and studies on wild-plant populations, we argue that studies on economic benefits from pollinators should include the total of ecosystem processes that (1) lead to successful pollination and (2) mobilize nutrients and improve plant quality to the extent that crop yields indeed benefit from enhanced pollinator services. Conservation incentives that use quantifications of nature's services to human welfare will benefit from approaches at the ecosystem level that take into account the broad spectrum of biological processes that limit or deliver the service. PMID- 17913146 TI - Development of analytical methods for polycyclic aromatic hydrocarbons (PAHs) in airborne particulates: a review. AB - In the present work, the different sample collection, pretreatment and analytical methods for polycyclic aromatic hydrocarbons (PAHs) in airborne particulates is systematacially reviewed, and the applications of these pretreatment and analytical methods for PAHs are compared in detail. Some comments on the future expectation are also presented. PMID- 17913147 TI - Inorganic nitrogen removal of toilet wastewater with an airlift external circulation membrane bioreactor. AB - Removal of inorganic nitrogen (inorganic-N) from toilet wastewater, using a pilot scale airlift external circulation membrane bioreactor (AEC-MBR) was studied. The results showed that the use of AEC-MBR with limited addition of alkaline reagents and volumetric loading rates of inorganic-N of 0.19-0.40 kg inorganic-N/(m3 x d) helped achieve the desired nitrification and denitrification. Furthermore, the effects of pH and dissolved oxygen (DO) on inorganic-N removal were examined. Under the condition of MLSS at 1.56-2.35 g/L, BOD5/ammonia nitrogen (NH4+ -N) at 1.0, pH at 7.0-7.5, and DO at 1.0-2.0 mg/L, the removal efficiencies of NH4+ -N and inorganic-N were 91.5% and 70.0%, respectively, in the AEC-MBR. The cost of addition of alkaline reagent was approximately 0.5-1.5 RMB yuan/m3, and the energy consumption was approximately 0.72 kWh/m3 at the flux of 8 L/(m2 x h). PMID- 17913148 TI - Aerobic degradation of methyl tert-butyl ether by a Proteobacteria strain in a closed culture system. AB - The contamination of methyl tert-butyl ether (MTBE) in underground waters has become a widely concerned problem all over the world. In this study, a novel closed culture system with oxygen supplied by H2O2 was introduced for MTBE aerobic biodegradation. After 7 d, almost all MTBE was degraded by a pure culture, a member of beta-Proteobacteria named as PM1, in a closed system with oxygen supply, while only 40% MTBE was degraded in one without oxygen supply. Dissolved oxygen (DO) levels of the broth in closed systems respectively with and without H2O2 were about 5-6 and 4 mg/L. Higher DO may improve the activity of monooxygemase, which is the key enzyme of metabolic pathway from MTBE to tert butyl alcohol and finally to CO2, and may result in the increase of the degrading activity of PM1 cell. The purge and trap GC-MS result of the broth in closed systems showed that tert-butyl alcohol, isopronol and acetone were the main intermediate products. PMID- 17913149 TI - Optimization of process parameters for the bioconversion of activated sludge by Penicillium corylophilum, using response surface methodology. AB - The optimization of process parameters for the bioconversion of activated sludge by Penicillium corylophilum was investigated using response surface methodology (RSM). The three parameters namely temperature of 33 degrees C, agitation of 150 r/min, and pH of 5 were chosen as center point from the previous study of fungal treatment. The experimental data on chemical oxygen demand (COD) removal (%) were fitted into a quadratic polynomial model using multiple regression analysis. The optimum process conditions were determined by analyzing response surface three dimensional surface plot and contour plot and by solving the regression model equation with Design Expert software. Box-Behnken design technique under RSM was used to optimize their interactions, which showed that an incubation temperature of 32.5 degrees C, agitation of 105 r/min, and pH of 5.5 were the best conditions. Under these conditions, the maximum predicted yield of COD removal was 98.43%. These optimum conditions were used to evaluate the trail experiment, and the maximum yield of COD removal was recorded as 98.5%. PMID- 17913150 TI - Distribution of platinum group elements in road dust in the Beijing metropolitan area, China. AB - Dust samples collected from the Beijing metropolitan area (China) were evaluated to determine the distribution and the concentration of platinum group elements (PGEs). The dust particles that were smaller than 100 mesh size fraction (150 microm) were analyzed after aqua regia digestion. Concentrations of Pt, Rh, and Pd were found to be between 3.96 and 356.3 ng/g, 2.76 and 97.11 ng/g, and 0.1 and 124.9 ng/g, respectively, in the urban areas of Beijing, whereas for the background samples collected from the suburbs of Beijing, the concentrations of Pt, Pd, and Rh were very low and ranged from 0.1 to 0.9 ng/g, 0.5 to 1.4 ng/g, and 0.8 to 2.2 ng/g, respectively. The distributions of PGEs in road dust were an accurate reflection of the levels of pollution and were found to match with the local traffic conditions. A strong positive correlation was established among all the elements found in road dust. This suggests that emissions of abraded fragments from vehicle exhausts may be the source of the high concentration of Pt, Rh, and Pd in road dust along the main roads of Beijing. PMID- 17913151 TI - Characteristics of indoor/outdoor PM2.5 and elemental components in generic urban, roadside and industrial plant areas of Guangzhou City, China. AB - Quantitative information on mass concentrations and other characteristics, such as spatial distribution, seasonal variation, indoor/outdoor (I/O) ratio, correlations and sources, of indoor and outdoor PM2.5 and elemental components in Guangzhou City were provided. Mass concentration of PM2.5 and elemental components were determined by standard weight method and proton-induced X-ray emission (PIXE) method. 18 elements were detected, the results showed positive results. Average indoor and outdoor PM2.5 concentrations in nine sites were in the range of 67.7-74.5 microg/m3 for summer period, and 109.9-123.7 microg/m3 for winter period, respectively. The sum of 18 elements average concentrations were 5362.6-5533.4 ng/m3 for summer period, and 8416.8-8900.6 ng/m3 for winter period, respectively. Average concentrations of PM2.5 and element components showed obvious spatial characteristic, that the concentrations in roadside area and in industrial plant area were higher than those in generic urban area. An obvious seasonal variation characteristic was found for PM2.5 and elemental components, that the concentrations in winter were higher than that in summer. The I/O ratio of PM2.5 and some elemental components presented larger than 1 sometimes. According to indoor/outdoor correlation of PM2.5 and element concentrations, it was found that there were often good relationships between indoor and outdoor concentrations. Enrichment factors were calculated to evaluate anthropogenic versus natural elements sources. PMID- 17913152 TI - Effects of exogenous salicylic acid on growth and H2O2-metabolizing enzymes in rice seedlings under lead stress. AB - Salicylic acid (SA) was an essential component of the plant resistance to pathogens and also plays an important role in mediating plant response to some abiotic stress. The possible effects of SA on the growth and H2O2-metabolizing enzymes in rice seedlings under lead stress were studied. When rice seedlings grown in nutrient solution containing Pb2+ (0, 0.05, 0.15, 0.25 mmol/L) for 18 d, the plant biomass as well as the chlorophyll content of leaves decreased with increasing Pb concentration. The pre-treatment with SA (treated with 0.1 mmol/L SA for 48 h before Pb stress) partially protected seedlings from Pb toxicity. The chlorophyll contents were significant higher in leaves of Pb-exposed with SA pre treatment seedlings than in Pb-exposed plants at the same Pb intensity. SA pre treated alone could significantly increase the length of shoot and root of seedlings but the vigour difference was not marked under long-term exposure to Pb toxicity. SA pre-treated influence the H202 level in leaves of seedlings by up regulating the activity of superoxide dismutase (SOD), repressing the activity of catalase (CAT) and ascorbate peroxidase (APX) depending on the concentrations of Pb2+ in the growth medium. The results supported the conclusion that SA played a positive role in rice seedlings against Pb toxicity. PMID- 17913153 TI - Assessment and mapping of environmental quality in agricultural soils of Zhejiang Province, China. AB - Heavy metal concentrations in agricultural soils of Zhejiang Province were monitored to indicate the status of heavy metal contamination and assess environmental quality of agricultural soils. A total of 908 soil samples were collected from 38 counties in Zhejiang Province and eight heavy metal (Cd, Cr, Pb, Hg, Cu, Zn, Ni and As) concentrations had been evaluated in agricultural soil. It was found 775 samples were unpolluted and 133 samples were slightly polluted and more respectively, that is approximately 14.65% agricultural soil samples had the heavy metal concentration above the threshold level in this province by means of Nemerow's synthetical pollution index method according to the second grade of Standards for Soil Environmental Quality of China (GB15618 1995). Contamination of Cd was the highest, followed by Ni, As and Zn were lower correspondingly. Moreover, Inverse Distance Weighted (IDW) interpolation method was used to make an assessment map of soil environmental quality based on the Nemerow's pollution index and the soil environmental quality was categorized into five grades. Moreover, ten indices were calculated as input parameters for principal component analysis (PCA) and the principal components (PCs) were created to compare environmental quality of different soils and regions. The results revealed that environmental quality of tea soils was better than that of paddy soils, vegetable soils and fruit soils. This study indicated that GIS combined with multivariate statistical approaches proved to be effective and powerful tool in the mapping of soil contamination distribution and the assessment of soil environmental quality on provincial scale, which is beneficial to environmental protection and management decision-making by local government. PMID- 17913154 TI - Phospholipid fatty acid patterns of microbial communities in paddy soil under different fertilizer treatments. AB - The microbial communities under irrigated rice cropping with different fertilizer treatments, including control (CK), PK, NK, NP, NPK fertilization, were investigated using phospholipid fatty acid (PLFA) profile method. The results of this study revealed that the fertilizer practice had an impact on the community structure of specific microbial groups. The principal components analysis (PCA) showed that proportion of the actinomycete PLFAs (10Me 18:0 and 10Me 16:0) were the lowest in the PK treatment and the highest in the NPK treatment, which means that soil nitrogen status affected the diversity of actinomycetes, whereas nitrogen cycling was related to the actinomycets. Under CK treatment, the ratio of Gram-positive to Gram-negative bacteria was lower compared with that in fertilizer addition treatments, indicating that fertilizer application stimulated Gram-positive bacterial population in paddy soil. The fatty acid 18:2omega6,9, which is considered to be predominantly of fungal origin, was at low level in all the treatments. The ratio of cyl9:0 to 18: 1omega7, which has been proposed as an indicator of stress conditions, decreased in PK treatment. Changes of soil microbial community under different fertilizer treatments of paddy soil were detected in this study; however, the causes that lead to changes in the microbial community still needs further study. PMID- 17913155 TI - Analysis of bacterial community structures in two sewage treatment plants with different sludge properties and treatment performance by nested PCR-DGGE method. AB - The bacterial community structures in two sewage treatment plants with different processes and performance were investigated by denaturing gradient gel electrophoresis (DGGE) of nested polymerase chain reaction (nested PCR) amplified 16S rRNA gene fragments with group-specific primers. Samples of raw sewage and treated effluents were amplified using the whole-cell PCR method, and the activated sludge samples were amplified using the extracted genomic DNA before the PCR products were loaded on the same DGGE gel for bacterial community analysis. Ammonia-oxidizing bacterial and actinomycetic community analysis were also carried out to investigate the relationship between specific population structures and system or sludge performance. The two plants demonstrated a similarity in bacterial community structures of raw sewage and activated sludge, but they had different effluent populations. Many dominant bacterial populations of raw sewage did not appear in the activated sludge samples, suggesting that the dominant bacterial populations in raw sewage might not play an important role during wastewater treatment. Although the two plants had different sludge properties in terms of settleability and foam forming ability, they demonstrated similar actinomycetic community structures. For activated sludge with bad settling performance, the treated water presented a similar DGGE pattern with that of activated sludge, indicating the nonselective washout of bacteria from the system. The plant with better ammonium removal efficiency showed higher ammonia-oxidizing bacteria species richness. Analysis of sequencing results showed that the major populations in raw sewage were uncultured bacterium, while in activated sludge the predominant populations were beta proteobacteria. PMID- 17913156 TI - Effects of cultivation conditions on the diversity of microbes involved in the conversion of rice straw to fodder. AB - To confirm the optimum cultivation conditions for analyzing lactic acid bacterial communities and to provide the cultivation foundation for lactic acid bacterial communities that were used to convert straw into fodder, fermented rice straw was inoculated into 13 different broths. After 48 h of cultivation, pH values, volatile products, and microbial diversity were analyzed. Except for LAB broth, the pH values of the other broths could decrease to approximately 4.5. GC/MS analysis showed that lactic acid in Tomato MRS broth, MRS broth, LAB broth, and Tomato juice broth was higher than that in the other broths. DNA concentration analysis showed that the counts of microbes in Tomato MRS broth were 2.5 times of those in other broths and that tomato juice favored the reproduction of the microbes. Denaturing gradient gel electrophoresis (DGGE) analysis showed that the number of lactic acid bacterial species in HYA broth, Tomato juice broth, and Tomato MRS broth were higher than those in the other broths. PMID- 17913157 TI - Bacterial diversity in soils around a lead and zinc mine. AB - Five samples of soil collected from a lead and zinc mine were used to assess the effect of combined contamination of heavy metals on soil bacterial communities using a polyphasic approach including characterization of isolates by culture method, community level catabolic profiling in BIOLOG GN microplates, and genetic community fingerprinting by denaturing gradient gel electrophoresis of 16S rDNA fragments amplified by PCR from community DNA (PCR-DGGE). The structure of the bacterial community was affected to a certain extent by heavy metals. The PCR DGGE analysis of 16S rRNA genes showed that there were significant differences in the structure of the microbial community among the soil samples, which were related to the contamination levels. The number of bacteria and the number of denaturing gradient gel electrophoresis (DGGE) bands in the soils increased with increasing distance from the lead and zinc mine tailing, whereas the concentration of lead (Pb) and cadmium (Cd) was decreased. Heavily polluted soils could be characterized by a community that differs from those of lightly polluted soils in richness and structure of dominating bacterial populations. The clustering analysis of the DGGE profiles showed that the bacteria in all the five samples of soil belonged to three clusters. The data from the BIOLOG analysis also showed the same result. This study showed that heavy metal contamination decreased both the biomass and diversity of the bacterial community in soil. PMID- 17913159 TI - Enhanced photocatalytic activity of TiO2 by surface fluorination in degradation of organic cationic compound. AB - Experiments were carried out to investigate the influence of TiO2 surface fluorination on the photodegradation of a representative organic cationic compound, Methylene Blue (MB). The electropositive MB shows poor adsorption on TiO2 surface; its degradation performs a HO-radical-mediated mechanism. In the F modified system, the kinetic reaction rate enlarged more than 2.5 fold that was attributed mainly to the accumulating adsorption of MB and the increased photogenerated hole available on the F-modified TiO2 surface. PMID- 17913158 TI - Preparations of TiO2 film coated on foam nickel substrate by sol-gel processes and its photocatalytic activity for degradation of acetaldehyde. AB - Anatase TiO2 films were successfully prepared on foam nickel substrates by sol gel technique using tetrabutyl titanate as precursor. The characteristics of the TiO2 films were investigated by XPS, XRD, FE-SEM, TEM and UV-Vis absorption spectra. The photocatalytic activities of TiO2 films were investigated by photocatalytic degradation reactions of gaseous acetaldehyde, an indoor pollutant, under ultraviolet light irradiation. It was found that Ni2+ doping into Ti02 films due to the foam nickel substrates resulted in the extension of absorption edges of TiO2 films from UV region to visible light region. The pre heating for foam nickel substrates resulted in the formation of NiO layer, which prevented effectively the injection of photogenerated electrons from TiO2 films to metal nickel. The TiO2 films displayed high photocatalytic activity for the degradation of acetaldehyde, and were enhanced by calcining the substrates and coating TiO2 films repeatedly. The high activity was mainly attributed to the improvement of the characteristics of substrate surface and the increase of active sites on photocatalyst. PMID- 17913160 TI - Preparation, characterization and photocatalytic activities of boron- and cerium codoped TiO2. AB - Boron- and cerium-codoped TiO2 photocatalysts were synthesized using modified sol gel reaction process and characterized by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), particle size distribution (PSD), diffuse reflectance spectra (DRS), and Brunauer-Emmett-Teller (BET). The photocatalytic activities were evaluated by monitoring the degradation of dye Acid Red B (ARB). The results showed that the prepared photocatalysts were mixed oxides mainly consisting of titania, ceria, and boron oxide. The structure of TiO2 could be transformed from amorphous to anatase and then to rutile by increasing calcination temperature; the transformation being accompanied by the growth of particle size without any obvious change in phase structure of CeO2. The XPS of B(1.6)Ce(1.0)-TiO2 prepared at 500 degrees C showed that a few boron atoms were incorporated into titania and ceria lattice, whereas others existed as B2O3. Cerium ions existed in two states, Ce3+ and Ce4+, and the atomic ratio of Ce3+/Ce4+ was 1.86. When boron and cerium were doped, the UV-Vis adsorption band wavelength showed an obvious shift toward the visible range (< or =526 nm). As the atomic ratio of Ce/Ti increased to 1.0, the absorbance edge wavelength increased to 481 nm. The absorbance edge wavelength decreased for higher cerium doping levels (Ce/Ti = 2.0). The particles size ranged from 122 to 255 nm with a domain at 168 nm (39.4%). The degradation of ARB dye indicated that the photocatalytic activities of boron- and cerium-codoped TiO2 were much higher than those of P25 (a standard TiO2 powder). The activities increased as the boron doping increased, whereas decreased when the Ce/Ti atomic ratio was greater than 0.5. The optimum atomic ratio of B/Ti and Ce/Ti was 1.6 and 0.5, respectively. PMID- 17913161 TI - Photocatalytic degradation of the dye sulforhodamine-B: a comparative study of different light sources. AB - The photocatalytic degradation of dye pollutant sulforhodamine-B (SRB) in aqueous titanium dioxide (TiO2) dispersions was examined under three lighting regimes: UV light (330 nm450 nm), all investigated at pH=2.5. Total organic carbon (TOC) and chemical oxygen demand (COD(Cr)) assays show that the degradation rate of SRB is much higher when irradiated with UV and sunlight compared with visible light. The temporal concentration changes of SRB illustrated a first-order reaction and the rate constant, k, is 0.197 min(-1), 0.152 min(-1), 0.027 min(-1), respectively, under the three lighting conditions. The final mineralized products were amine compounds identified by infrared spectrophotometry. When irradiated with visible light, the photocatalytic degradation rate could be improved by lowering the H202 concentration and inhibited by increasing the H2O2 concentration, but results contrary to the above were obtained when UV light was used for irradiation. PMID- 17913163 TI - Emissions of SO2, NO and N2O in a circulating fluidized bed combustor during co firing coal and biomass. AB - This paper presents the experimental investigations of the emissions of SO2, NO and N20 in a bench scale circulating fluidized bed combustor for coal combustion and co-firing coal and biomass. The thermal capacity of the combustor is 30 kW. The setup is electrically heated during startup. The influence of the excess air, the degree of the air staging, the biomass share and the feeding position of the fuels on the emissions of SO2, NO and N2O were studied. The results showed that an increase in the biomass shares resulted in an increase of the CO concentration in the flue gas, probably due to the high volatile content of the biomass. In co firing, the emission of SO2 increased with increasing biomass share slightly, however, non-linear increase relationship between SO2 emission and fuel sulfur content was observed. Air staging significantly decreased the NO emission without raising the SO2 level. Although the change of the fuel feeding position from riser to downer resulted in a decrease in the NO emission level, no obvious change was observed for the SO2 level. Taking the coal feeding position R as a reference, the relative NO emission could significantly decrease during co-firing coal and biomass when feeding fuel at position D and keeping the first stage stoichiometry greater than 0.95. The possible mechanisms of the sulfur and nitrogen chemistry at these conditions were discussed and the ways of simultaneous reduction of SO2, NO and N20 were proposed. PMID- 17913162 TI - Activated carbons derived from oil palm empty-fruit bunches: application to environmental problems. AB - Activated carbons derived from oil palm empty fruit bunches (EFB) were investigated to find the suitability of its application for removal of phenol in aqueous solution through adsorption process. Two types of activation namely; thermal activation at 300, 500 and 800 degrees C and physical activation at 150 degrees C (boiling treatment) were used for the production of the activated carbons. A control (untreated EFB) was used to compare the adsorption capacity of the activated carbons produced from these processes. The results indicated that the activated carbon derived at the temperature of 800 degrees C showed maximum absorption capacity in the aqueous solution of phenol. Batch adsorption studies showed an equilibrium time of 6 h for the activated carbon at 800 degrees C. It was observed that the adsorption capacity was higher at lower values of pH (2-3) and higher value of initial concentration of phenol (200-300 mg/L). The equilibrium data fitted better with the Freundlich adsorption isotherm compared to the Langmuir. Kinetic studies of phenol adsorption onto activated carbons were also studied to evaluate the adsorption rate. The estimated cost for production of activated carbon from EFB was shown in lower price (USD 0.50/kg of activated carbon) compared the activated carbon from other sources and processes. PMID- 17913164 TI - Experimental and modeling study of de novo formation of PCDD/PCDF on MSW fly ash. AB - The effect of sulfur dioxide (SO2) on the formation of polychlorinated dibenzo-p dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) has been studied in an entrained-flow reactor (EFR) under simulated waste combustion conditions. A chlorination model based on conditional probability was employed to evaluate the homologue patterns of PCDDs and PCDFs. Results revealed that the presence of SO2 did not alter the formation pathway although SO2 suppressed PCDD/F formation. The prediction model of PCDF showed good agreement with the experimental data (R = 0.95), whereas the prediction for PCDDs did not correlate well with the experimental data. This may be explained because potential chlorination pathways play a significant role in PCDF formation, whereas PCDDs are mainly formed through condensation reactions. Furthermore, the result indicated that the steric hindrance during formation has more effects on PCDD than on PCDF due to the symmetric molecular structures of PCDDs. PMID- 17913165 TI - Sediment-copper distributions in hyper-concentrated turbulent solid-liquid system. AB - This study presents a special problem on vertical distribution for sediment and copper in hyper-concentrated turbulent solid-liquid system that is essentially different from the ordinary low-concentrated turbulent system. A resonance type turbulent simulation equipment is used for the experimental study in which a vertically uniform turbulent field of the mixture of loess and water is produced in a testing cylinder with a grille stirrer that moves up and down harmoniously with varying vibration frequencies. In order to compare the variations of the vertical profiles of sediment and copper in low- and hyper-concentrated solid liquid system, different scenarios for input sediment content ranging from 5 to 800 kg/m3 was considered in the experimental studies. It was found that solids copper content increases with input sediment content, S0, and reaches its peak as S0 goes to 10 kg/m3 and then decreases rapidly with increasing input sediment content. Such a behavior is possibly resulted from the joint effect of the specific adsorption of copper on loess, precipitation of carbonate and hydroxide of copper due to high carbonate content in the loess and the so-called "particulate concentration effect" due to the present of the sediment variation in water. The vertical sediment concentration distribution resulted from the uniform turbulence is generally uniform, but slight non-uniformity does occur as sediment concentration exceeds certain value. However, the vertical concentration distributions of soluble copper seem not to be affected much by the variation of sediment concentrations. PMID- 17913166 TI - Orbital myositis in a child with linear scleroderma en coup de sabre. PMID- 17913167 TI - Acute comitant esotropia. PMID- 17913168 TI - What's your diagnosis? Acquired and progressive myelinated nerve fibers. PMID- 17913169 TI - Malignant melanoma of the conjunctiva in children: a review of the international literature 1965-2006. AB - PURPOSE: Conjunctival melanoma is rare in children. This article presents a comprehensive review and discusses the clinical significance of the published cases of malignant melanoma of the conjunctiva in children younger than 15 years. MATERIALS AND METHODS: To obtain prior reports of conjunctival melanoma in children, a systematic search of the world literature was conducted. Additional studies were identified from the bibliographies of the retrieved articles, as well as from major ophthalmic textbooks. RESULTS: A review of the literature resulted in 28 reported cases of conjunctival melanoma in children younger than 15 years, and only 8 had adequate case details. CONCLUSION: Malignant melanoma of the conjunctiva is a rare condition in children. Limited knowledge for management and prognosis of this condition in children is largely due to its low occurrence. PMID- 17913170 TI - Effectiveness of disinsertion-resection and tucking of the inferior oblique muscle in patients with unilateral long-standing superior oblique muscle palsy. AB - PURPOSE: To investigate the effectiveness and safety of disinsertion-resection and tucking of the inferior oblique muscle in patients with unilateral long standing superior oblique muscle palsy and secondary inferior oblique muscle overaction. METHODS: Between April 2000 and January 2005, the records of 31 patients who underwent disinsertion-resection and tucking of the inferior oblique muscle for treatment of unilateral long-standing (> 6 months) superior oblique muscle palsy were retrospectively reviewed. A comprehensive ocular examination including best-corrected visual acuity measurements, ductions, versions, and deviations at near and distance in the diagnostic positions of gaze, head tilt test, abnormal head position, dilated fundus, field of binocular fixation, and Lee screen test was performed prior to and after surgery. RESULTS: All patients had Knapp's class I unilateral superior oblique muscle palsy. The mean preoperative score of inferior oblique muscle overaction was +3.03 and the mean vertical deviation was 15.9 PD in primary position. The follow-up period ranged from 4 to 82 months. Inferior oblique muscle overaction diminished in 29 patients, and 2 patients had +1.0 overaction in adduction of the affected eye. The vertical deviation in these patients had some residual but smaller hypertropia. CONCLUSIONS: Disinsertion-resection and tucking of the inferior oblique muscle was safe, simple, and effective in eliminating inferior oblique muscle overaction and abnormal head posture, and in reducing the hyperdeviation in patients with unilateral long-standing superior oblique muscle palsy. PMID- 17913171 TI - Pars planitis in children: epidemiologic, clinical, and therapeutic characteristics. AB - PURPOSE: To report the demographics and clinical characteristics, therapy logarithm, and prognosis of children with pars planitis. PATIENTS AND METHODS: The medical records were reviewed of all patients diagnosed with pars planitis between June 1995 and December 2005 in the Department of Pediatric Ophthalmology at Hospital Universitario La Paz, Madrid, Spain. A retrospective, descriptive, and longitudinal study of 30 eyes in 16 children was performed. RESULTS: Pars planitis was bilateral in 87.5% and more frequent in males (68.8%). Average age at onset was 9.2 years. The main ophthalmologic findings recorded were snowballs (96.7%) and vitritis (93.3%). Cataract formation was the most prevalent complication (36.7%). Mean initial and final best-corrected visual acuities were 0.640 and 0.840, respectively. Periocular corticosteroids were used in 33.3% of cases and cryotherapy or laser photocoagulation in 16.7%. Complications requiring surgical management occurred in 4 eyes (13.3%). CONCLUSION: Pars planitis treated with adequate medical and surgical procedures has a good prognosis in most cases. PMID- 17913172 TI - Prevalence of refractive error and other eye diseases in schoolchildren in the Greater Accra region of Ghana. AB - PURPOSE: To determine the prevalence of refractive error and other eye diseases in schoolchildren in the Greater Accra region of Ghana. METHODS: Research assistants tested visual acuity (VA) using a Snellen chart in students randomly selected from 24 schools (i.e., 8 metropolitan, 8 urban, and 8 rural). An ophthalmologist examined students with VA of 6/12 or worse in either eye and students with other eye problems. RESULTS: Nine hundred fifty-seven students aged 6 to 22 years were screened: 67 had myopia with VA of 6/12 or worse in the better eye, 3 were unilaterally blind (2 from squint or amblyopia, 1 from a hypoplastic disk), and 4 had unilateral low vision from macular scars. Other eye diseases observed were congenital glaucoma (1), vernal conjunctivitis (2), and infective conjunctivitis (6). The prevalence of refractive error reducing vision to at least 6/12 was 7.0%. The prevalence of squint and amblyopia was 0.2%. CONCLUSION: The most common eye defect in the students was refractive error. Squint and amblyopia were rare. PMID- 17913173 TI - Conjunctival melanoma 3 years after radiation and chemotherapy for retinoblastoma. AB - An 8-month-old infant with advanced bilateral retinoblastoma was managed with enucleation of the right eye and chemotherapy and radiotherapy of the left eye. Three years after treatment, an ill-defined, nonpigmented mass was noted in the anophthalmic right socket. Excisional biopsy revealed an amelanotic melanoma of the conjunctiva with superficial orbital invasion. There has been no tumor recurrence during 20 months of follow-up. PMID- 17913174 TI - Sudden unilateral blindness in a girl with Kawasaki disease. AB - This article describes a 9-year-old girl with clinical and laboratory features of Kawasaki disease. On day 7 of her febrile illness, she developed sudden loss of vision in the right eye. Visual acuity was no light perception. Dilated funduscopy showed diffuse intense retinal whitening, narrowing retinal arterioles, and a pale swollen disk-but no cherry-red spot. These findings suggested ophthalmic artery obstruction. Kawasaki disease can be sight threatening. PMID- 17913175 TI - Fundus albipunctatus in a patient with achondroplasia. AB - We describe a patient with achondroplasia who presented with defective night vision due to fundus albipunctatus. We found no previous report of the association in a search of the literature. PMID- 17913176 TI - Nonsurgical management of an intraorbital pencil without cranial or globe penetration. AB - A 3-year-old boy suffered a penetrating orbital pencil injury in the right upper eyelid. Examination showed no signs of globe perforation, fractures, or neurologic deficits. The pencil was removed with controlled and slow motion. Visual acuity was 20/20 and anterior and posterior segment findings were normal 2 months after the injury. PMID- 17913177 TI - Neisseria meningitidis endogenous endophthalmitis presenting as anterior uveitis. AB - This article describes an unusual case of meningococcal endogenous endophthalmitis presenting as anterior uveitis. This case re-emphasizes the importance of early diagnosis of endogenous endophthalmitis, which can lead to good visual recovery. PMID- 17913178 TI - Atypical synophthalmos with arhinencephaly unilateralis. AB - This article describes a rare case of a 4-month-old infant with atypical synophthalmos with ipsilateral arhinencephaly unilateralis, ethmoid sinus, and lacrimal apparatus and brain abnormalities--diagnosed on the basis of the clinical picture and imaging findings. PMID- 17913180 TI - Siderosis bulbi in a child. PMID- 17913179 TI - Traumatic and spontaneous scleral rupture and uveal prolapse in osteogenesis imperfecta. AB - Three cases of severe globe injuries due to scleral fragility in osteogenesis imperfecta patients between the ages of 4 and 15 years are reported. Patient 1 had complete loss of vision. Patients 2 and 3 suffered non-sight-threatening scleral perforation. All 3 patients had no previous knowledge of recommendation for eyewear protection. PMID- 17913181 TI - Study of hydrodynamic behavior of a laboratory scale upflow anaerobic fixed film fixed bed reactor. AB - Application of anaerobic fixed film fixed bed reactor (AFFFB), containing mixed population of bacteria, helps in bacterial immobilization as biofilm on a support medium in the reactor overcoming the problem of biomass washout. Knowledge of hydrodynamics characteristics helps in better understanding and evaluating the performance of any reactor. This paper presents residence time distribution (RTD) studies carried out to investigate the hydrodynamic behavior of AFFFB reactor under the influence of varied hydraulic loading resulting in HRTs in range 24 hrs to 6 hrs. Results have demonstrated that the flow pattern is more close to a plug flow type and vary with varying intensity of dispersion and the extent of dispersion increases with decrease in HRTs. PMID- 17913182 TI - Impacts of mining activities on water and soil. AB - Seven coal mines are situated in Wardha River Valley. These mines are located at Wani (Dist. Yavatmal of Maharashtra). Out of these, 5 open cast coal mines are run by Western Coal Field Ltd. India. The present study has been undertaken to assess the impacts of mining activities in the adjacent areas. Total 25 samples of water and 19 samples of soil from Nilapur, Bramhani, Kolera, Gowari, Pimpari and Aheri were analyzed for pH, TDS, hardness, alkalinity, fluoride, chloride, nitrite, nitrate, phosphate, sulfate, cadmium, lead, zinc, copper, nickel, arsenic, manganese, sodium and potassium, and the results were compared with the limits of Indian Standards: 10500. PMID- 17913183 TI - Assessment, management and spatial distribution of ground water for irrigation in Rupnagar district of Punjab (India). AB - Ground water samples for geo-referenced sites were collected and analyzed for various chemical constituents for assessing their quality for irrigation. These waters were categorized as good and marginal sodic based upon the values of EC and RSC. The ground water quality map of the area was prepared and digitized using GIS software and blockwise area under different categories was commuted, which indicates that Kharar block (46.7 percent of the TGA) has maximum area under marginally sodic water as compared to the Anandpur Sahib block with 8.5 percent of its TGA under marginally sodic waters. PMID- 17913184 TI - Studies on exhaust emissions of catalytic coated spark ignition engine with adulterated gasoline. AB - Adulteration of automotive fuels, especially, gasoline with cheaper fuels is widespread throughout south Asia. Some adulterants decrease the performance and life of the engine and increase the emission of harmful pollutants causing environmental and health problems. The present investigation is carried out to study the exhaust emissions from a single cylinder spark ignition (SI) engine with kerosene blended gasoline with different versions of the engine, such as conventional engine and catalytic coated engine with different proportions of the kerosene ranging from 0% to 40% by volume in steps of 10% in the kerosene gasoline blend. The catalytic coated engine used in the study has copper coating of thickness 400 microns on piston and inner surface of the cylinder head. The pollutants in the exhaust, carbon monoxide (CO) and unburnt hydrocarbons (UBHC) are measured with Netel Chromatograph CO and HC analyzer at peak load operation of the engine. The engine is provided with catalytic converter with sponge iron as a catalyst to control the pollutants from the exhaust of the engine. An air injection is also provided to the catalytic converter to further reduce the pollutants. The pollutants found to increase drastically with adulterated gasoline. Copper-coated engine with catalytic converter significantly reduced pollutants, when compared to conventional engine. PMID- 17913185 TI - Analytical study of heavy metals of industrial effluents at Jaipur, Rajasthan (India). AB - Heavy metals contamination has been recognized as a major environmental concern due to their pervasiveness and persistence. These heavy metals are not biodegradable, hence there is a need to develop such a remediation technique, which should be efficient, economical and rapidly deployable in a wide range of physical settings. For the characterization of heavy metals of various industrial effluents, some heavy metals, like Arsenic, Cadmium, Chromium, Copper, Iron, Manganese, Nickel, Lead and Zinc were analyzed. The results exhibited that As, Cd, Cr and Pb were not found in any studied wastewater samples, while some of the following heavy metals ranged from: Cu (0.0 - 1.0 mg/L), Fe (0.1 - 0.4 mg/L), Mn (0.0 - 0.4 mg/L), Ni (0.01 - 0.07 mg/L) and Zn (0.68 - 60.84 mg/L). Copper, Iron, Manganese and Zinc were found above the standard limit recommended by IS: 3307 (1977). However, Nickel was found below the regulated safety values for all studied samples. PMID- 17913186 TI - A study on sludge minimization during the treatment of pickling effluent. AB - In pickling industries, a lot of sludge is generated during the treatment of pickling effluent and there is severe problem of its disposal. Disposal of this sludge as per the Hazardous Waste (Management & Handling) Rules, 1989 is not easy. Its transportation and construction of lined disposal sites pose very severe problems. In the normal practice, the sludge is being disposed of at the sides of roads and railway tracks to fill low lying areas. This may cause serious health hazards. Considering these problems, a study has been undertaken to minimize the sludge generation during the treatment of pickling effluent by neutralizing it with lime, sodium hydroxide and combination of both. An attempt has been made to do an economic evaluation of the above process. PMID- 17913187 TI - Evaluation of pollution susceptibility of Karst aquifers of Rewa Town (Madhya Pradesh) using "DRASTIC" approach. AB - Pollution susceptibility of groundwater of Rewa town situated on karstified Bhander limestones of the Bhander group is discussed in this paper. Pollution potential of selected localities in the town has been determined using the DRASTIC INDEX methodology. Pollution potential for these localities varied between 162 to 217. Shallow aquifers in karstified limestones having direct access to surface water were found more susceptible to pollution. Accordingly, remedial measures were suggested for minimising pollution. PMID- 17913188 TI - Assessment of coastal pollution through bioassay and transplantation of intertidal clams. AB - In the present study, a survey of Mahim creek and Bay area was undertaken that indicated absence of fauna, particularly molluscs from the area, which was a repository in the past. During bioassay experiments of Mahim creek water, the clams G. divaricatum and C. antiquata could not open their valves in 100% creek water and died within 12 hrs of exposure. The 96 hrs LC50 values of Mahim creek water for G. divaricatum and C. antiquata were found to be 20% and 40% respectively during summer and 38% and 57% respectively during rainy season. When two sets of the clams were transplanted at Mahim creek, they died within 12 hrs. These experiments suggest the extreme level of pollution in the area. This level of pollution is responsible for transforming the area into barren locality in terms of fauna, specially the rich molluscan diversity. However, Gorai creek was found to be comparatively very less polluted and it still serves as breeding and nursery ground for various fishes and prawn species. Hence it must be protected and conserved. PMID- 17913189 TI - Dynamics of lead (Pb) in different soil conditions. AB - Four soil samples were collected from various locations having wider range of pH and EC for incubation experiment. These soil samples were added with Pb (NO3)2 salts, and samples were taken on 1st, 35th, 45thand 60th day. Soils samples were fractioned by sequential extraction to estimate the concentration of lead in different fractions viz, water soluble, exchangeable +adsorbed, organic, carbonate and residual fraction. The result reveals that concentration of organic and carbonate bound-Pb was high in waterlogged condition and soil pH has been comprehensively identified as the single most important soil factor controlling the availability of lead (Pb) in soil. Low content of Pb in exchangeable + adsorbed (KNO3) and water soluble (H2O) fraction in all soils (except in S1) could signify low availability of Pb to plants. Bioavailable fractions, viz. water soluble and exchangeable + adsorbed, were low in all soils (except S1) well below critical limits, which may not pose any toxicity in the food chain. PMID- 17913190 TI - Monitoring of fluoride concentration in ground water of Prakasham District in India: correlation with physico-chemical parameters. AB - Fluoride concentrations in surface and ground water samples were determined in eight villages of Prakasham district in India. Thirty-eight samples were collected and analysed for fluoride content along with pH, electrical conductivity, total dissolved solids (TDS), total hardness, total alkalinity, chlorides (C1), sulfates (SO2-) and nitrates (NO ). Fluoride concentrations in surface and ground water samples of these villages varied between 0.5 and 9.0 mg/L. Groundwater samples contained high concentrations of fluorides compared to open well and pond water samples, which could be a major source of fluoride in water since the geological formation of this area consists of fluorite and fluoropatite. From the data, it is evident that the population in the study area is severely affected by fluorosis. Dental and skeletal fluorosis and deformation of bones in children as well as adults were observed in the study area indicating the consequences of excess fluoride concentration. Fluoride concentrations showed good correlation with TDS concentrations (R2 of 0.61) compared to other physico chemical parameters [EC (R2 - 0.36), nitrate (R - 0.24), total hardness (R - 0.12), chloride (R2 - 0.06) and sulfate (R2 - 4 x 10(-6))]. PMID- 17913191 TI - Removal of fluoride from water with powdered corn cobs. AB - The adsorption of fluoride on corn cobs powder was investigated in the present study. Neat powdered corn cobs did not show remarkable adsorption but aluminium treated corn cobs had good adsorption capacity. The parameters studied include the contact time, concentration, temperature and pH. Near neutral pH was identified as the optimum condition of the medium, and 90 to 120 minutes was the best contact time for maximum fluoride adsorption. The adsorption process was found to follow Freundlich isotherm. The adsorption process was found to be exothermic as adsorption decreased with increasing temperature. PMID- 17913192 TI - Noise pollution levels in Visakhapatnam City (India). AB - Visakhapatnam is an industrial and sea port city located on the east coast of India. A hospital (RCD hospital), residential area (Lawson's Bay Colony), traffic zone (Jagadamba junction, Andhra Pradesh State Road Transport Corporation Complex junction and Seethammadhara junction) and industrial zone (Sea port) were chosen to monitor the noise levels. The observed noise level at RCD hospital was more than 10 dBA in any time. The background noise at Santhi Ashram was approximately 3dBA less at night time and 2dBA less at day time compared to ambient air quality noise standards (AAQNS) for silent zone. The ambient air quality noise levels (AAQNL) at traffic junctions were 5 dBA or more than those prescribed by AAQNS for commercial zone and most of the values were found in the range of 80 +/- 10 dBA, among which 75% values were found in the range of 110 +/- 10 dBA. AAQNL near port were found in the range of 5 to 10 dBA positive shifts on AAQNS due to conveyor operation. The AAQNL were alarming even in the absence of conveyor system, indicating the impact of vehicular traffic. Remedial measures were suggested separately for each situation. PMID- 17913193 TI - Environmental management of industrial hazardous wastes in India. AB - Hazardous wastes are considered highly toxic and therefore disposal of such wastes needs proper attention so as to reduce possible environmental hazards. Industrial growth has resulted in generation of huge volume of hazardous wastes in the country. In addition to this, hazardous wastes sometimes get imported mainly from the western countries for re-processing or recycling. Inventorisation of hazardous wastes generating units in the country is not yet completed. Scientific disposal of hazardous wastes has become a major environmental issue in India. Hazardous Wastes (Management and Handling) Rules, 1989 have been framed by the Central Government and amended in 2000 and 2003 to deal with the hazardous wastes related environmental problems that may arise in the near future. This paper gives details about the hazardous wastes management in India. Health effects of the selected hazardous substances are also discussed in the paper. PMID- 17913194 TI - Modelling of secondary clarifier using regression analysis and artificial neural networks. AB - Mathematical models for the surface area of secondary clarifier were developed for wastewater generated from a dairy industry and from domestic sources, by correlating the parameters namely, surface area per unit flow rate (A/Q), influent concentration (C(O)), underflow concentration (C(U)), recycling ratio (r) and Mean Cell Residence Time (theta C) using multiple regression analysis. There was found a good correlation between the measured data and the model results with regression coefficients of 0.9. Thickener area requirement of combined wastewater was comparehat obtained for dairy wastewater. Thickener area was found to decrease with increase in Mean Cell Residence Time and the area required for treating the combined wastewater was less, when compared with the requirement for dairy wastewater treatment. Neural network was trained with experimental data to 'acquire' knowledge about it. The Back Propagation Network technique was used in which the error was back propagated through the network. The results evolved from the neural network training were compared with the results of regression model and experimental data. Greater deviation was observed between the observed and predicted values of A/Q at high underflow concentrations, indicating that the limiting solids flux was reached. The output from Neural Network approach had greater consistency with the experimental data than the output from conventional regression analysis. Hence, Artificial Neural Network technique is highly adaptive and efficient in investigating input - output relationships. PMID- 17913195 TI - A novel eco-friendly approach for textile sludge. AB - Solid waste management is a very serious problem in both human settlements and industrial establishments. In such cirumstances, management of sludge in textile industry has now become a burning issue due to its surplus volume and undesired characteristics. In such grim scenario, an attempt has been made to effectively dispose of the sludge generated in the treatment units of the textile sectors of Tirupur (India). The main objective of this research work was to study the suitability of utilizing sludge in the manufacture of bricks. It thus serves two important purposes, namely - efficient management of textile sludge and production of useful by-product, which fetches cash to the industry and a low cost construction material to the public. The sludge was collected from the effluent treatment plant and it was analyzed to explore its characteristics. Then the bricks, made up of this sludge, were tested for water absorption, compressive strength, presence of soluble salts and drop test, and the results were compared with those of ordinary bricks. PMID- 17913196 TI - Trace metal levels in the organs of finfish Oreochromis mossambicus (Peter) and relevant water of Jannapura lake, India. AB - Trace metal levels in the body organs of finfish, Oreochromis mossambicus (Peter) and relevant water was studied in Jannapura lake, located five kilometers from Bhadravathi town, Karnataka, India, during 2004-05. Lead, Copper and Cadmium accumulation was higher in muscles than in gills while, Zinc, Nickel and Cobalt accumulation was maximum in gills followed by muscles. The metals present in the highest concentration were in the order of Pb > Cu > Zn > Cd > Ni > Co in the water samples. The higher concentration of various metals in water and Pb, Ni in fish muscle and Ni in gill samples was probably due to the addition of untreated sewage, geological weathering and agricultural runoff from the surrounding areas. On the basis of results obtained, it is predicted that if the present situation of water pollution continues in future, survival of fish population and other aquatic animals will be extremely difficult. PMID- 17913197 TI - Modelling of gaseous effluents by implementing Gaussian model under GIS environment. AB - In the present work, Gaussian based air quality model has been implemented under GIS environment for the air quality emission data obtained from IFFCO power plant at Phulpur, Allahabad, India. The application of corrections for the non-point source, obstacles on ground, surface roughness and observation time has made the Gaussian based air pollution model more accurate and flexible for its use and in its adaptation for any other location. The Gaussian based air pollution model for the IFFCO plant, implemented under GIS environment, has been tested for various validation points to check its efficacy and has been found to be suitable for prediction of air pollutants in future. Further, a user interactive modelling interface has been developed using Visual Basic as a front-end, implementing the ArcObjects of the ArcGIS 8.3 to make it more user-friendly and increase its acceptability among environmentalists, planners and decision makers. PMID- 17913198 TI - Major ion and heavy metal chemistry of Pachin river (Itanagar) - levels and sources. AB - The Pachin river is an upland tributary of the Brahmaputra river, originating in the foot-hills of the lesser Himalayas. A systematic study of major ions and heavy metals in the surface water of the river was carried out at peak, intermediate and low flow conditions during an one-year interval to assess the relative contributions from weathering and pollution. The major ion chemistry indicates that silicate weathering and precipitation are the major contributing factors to the river's chemical composition. The Pachin river is characterized by a low overall conductivity, even during times of evaporative concentrations during low flow. This corresponds to the fact that the catchment area is still relatively pristine, i.e., well forested and without significant industry. However, heavy metal patterns clearly reflect source input from agricultural activity and urban development. The levels of Fe and Co were amongst the highest of any rivers of the Indian sub-continent, while these and Cr, Mn, Cu and Se each exceeded the world average value. These results clearly indicate that rivers draining pristine areas can be significantly modified by human activities, and consequently, these must be monitored to ensure that useable water supplies meet the prescribed safety standards. PMID- 17913199 TI - A study of noise around an educational institutional area. AB - Road transportation has played a major role in the economic and social development of our country. Vehicles such as two and four wheelers are preferred as they provide quick means of door to door service to the community. With increase in the vehicular population, the noise levels are also increasing. The present paper discusses the results of a study undertaken to assess the noise levels at the major traffic junctions and community area near an educational institution of an urban city. Noise equivalent level Leq was measured in the neighborhood community areas as well as at the traffic junctions. The statistical levels L10, L50, L90 were also measured to determine the Traffic Noise Index at the junctions. A holistic approach was adopted to study the impact of noise on the community. The study indicates a need for proper land-use planning when traffic corridors are built in the silence zone areas. PMID- 17913200 TI - Monthly variation in night time noise levels at residential areas of Asansol city (India). AB - Intense vehicular movement and industrial activities are the major noise producing sources in the Asansol city. Ambient night time noise levels were monitored in ten residential areas from August 2002 to July 2003, to study the existing quality of noise and their impact in the local community. Based on the monitored noise levels in the area, traffic-social survey was also conducted. About 39% of individuals surveyed were highly annoyed with noise pollution from different origins. The study revealed that night time noise levels (10.00 pm - 6.00 am) in all the locations exceeded the limit prescribed by Central Pollution Control Board. The day time noise level was much higher at all locations in respect of the night time noise level. The Day-Night equivalent noise level (Ldn) was determined and ranged between 67.16 dB(A) and 89.44 dB(A). The percentile noise level (L90) exceeded the CPCB standard in all cases and varied from 72.36 dB(A) to 102.45 dB(A). Based on the results of the study, it can be concluded that the local community is exposed to high noise environment. PMID- 17913201 TI - Gamma radiation process for destruction of toxic polychlorinated biphenyls (Pcbs) in transformer oils. AB - Polychlorinated biphenyls (PCBs) are synthetic organic chemicals commercially used worldwide in many applications. PCBs were used in oils because of their excellent properties such as good thermal stability, flame resistance, dielectric constant, high break down voltage, high boiling point and low volatility. However, because of their adverse affects on environment and human health, the use of PCBs has been banned now. PCBs are today considered among the widespread pollutants in the global system. PCBs sources still exist in various industrial products and in waste streams such as capacitor oils, lubricating oils, transformer oils, hydraulic oils, paints, rubbers, cables, etc. Several such materials containing PCBs emanating from various sources need to be detoxified before their reuse or before going to landfill for final disposal. Various remedial technologies have been developed in the world to destroy toxic PCBs. The radiolysis has been investigated as an environment-friendly process for waste oil treatment contaminated with PCBs, which may be a better alternative to the globally most widely accepted incineration method. A study was undertaken to detoxify PCBs in transformer oil by gamma radiation using Cobalt 60 source. Analysis of PCBs in transformer oils before and after radiation was carried out by GC-MS instrument. The effect of radiation dose and destruction of PCBs in transformer oils are discussed in details in the present paper. The method used was found to be highly effective and destruction was as high as 79 %. Further, the transformer oil samples were also evaluated before and after radiation to check their quality. The properties of oils were not significantly altered by gamma radiation treatment as evident from the results given in the paper. PMID- 17913202 TI - A study of electro-mechanical and infrastructure instrumentation facilities in environmental laboratory. AB - Environmental laboratories carry out measurement and analysis of a number of physical, chemical and biological parameters. Each parameter requires some sort of instrument for its determination. Providing efficient instrumentation services to various departments of the Institute is an stupendous task. Instrumentation services in the form of installation, operation, repair and maintenanace of electro-mechanical equipment requires an in-depth experience and knowledge of the working, fabrication, design and repair of similar type of instruments so that the need of space, installation pre-requisites, budget constraints, availability of essential spares parts could be assessed. The paper discusses the operation of an environmental instrument repairs and maintenance, and audio-visual facilities. Suggestions for drafting of the proper specifications for procurement of laboratory equipments, such as ovens, furnaces, refrigerators, blowers, audio visual aids, and spares and accessories are given in this paper. The paper also gives the detailed information on various aspects that are needed for checking and testing of the equipment against specification before putting it in operational use. Development of a preventive maintenance program involving QC checks and keeping an inventory of essential spares required are also discussed in this paper. It is felt that such services are essential in providing smooth support to carry out research and development activities of the Institute. PMID- 17913203 TI - Improvement of water quality through biological denitrification. AB - The present study was undertaken to observe the effect of biological denitrification process on water quality parameters. The results obtained in the present investigations show that the value of pH and alkalinity was increased due to generation of alkalinity during biological denitrification process. The obtained value of the DO in the treated water was found lower than the supplied water, which indicates, DO was consumed by the bacterium. The COD of the treated water was nil, which shows that the organics have been consumed by the bacterium during the denitrification process. The biological reduction of nitrate-nitrogen (from 50.79 mg/L to 0.57 mg/L) was found to be lower than the tolerance limit prescribed by WHO without changing the water quality. PMID- 17913204 TI - Control of aldehyde emissions in the diesel engines with alcoholic fuels. AB - The major pollutants emitted from compression ignition (CI) engine with diesel as fuel are smoke and nitrogen oxides (NOx). When the diesel engine is run with alternate fuels, there is need to check alcohols (methanol or ethanol) and aldehydes also. Alcohols cannot be used directly in diesel engine and hence engine modification is essential as alcohols have low cetane number and high latent hear of vaporization. Hence, for use of alcohol in diesel engine, it needs hot combustion chamber, which is provided by low heat rejection (LHR) diesel engine with an air gap insulated piston with superni crown and air gap insulated liner with superni insert. In the present study, the pollution levels of aldehydes are reported with the use of methanol and ethanol as alternate fuels in LHR diesel engine with varying injection pressure, injection timings with different percentage of alcohol induction. The aldehydes (formaldehyde and acetaldehyde) in the exhaust were estimated by wet chemical technique with high performance liquid chromatograph (HPLC). Aldehyde emissions increased with an increase in alcohol induction. The LHR engine showed a decrease in aldehyde emissions when compared to conventional engine. However, the variation of injection pressure showed a marginal effect in reducing aldehydes, while advancing the injection timing reduced aldehyde emissions. PMID- 17913205 TI - Study of fluoride in ground water around the BALCO, Korba area (India). AB - The present study was undertaken for the determination of fluoride ions in drinking water at the BALCO, Korba region by the ion selective electrode method. The ground water samples of thirty four villages of this region were collected. The fluoride concentration values varied from 1.07 ppm to 3.10 ppm. In this area, villagers were found suffering with dental and skeletal fluorosis. The villagers were suggested to use alum, lime and Nalgonda technique to avoid all type of fluorosis. It was found that fluoride was present within the permissible limit (1.5 ppm) in most of the villages studied but the fluoride level was unacceptable in drinking water samples taken from some villages of BALCO, Korba area. PMID- 17913206 TI - Adsorption of anionic dyes on to waste Fe (III)/Cr (III). AB - Waste Fe (III)/Cr (III) hydroxide was investigated for the removal of anionic dyes, namely acid brilliant blue (acidic dye) and procion red (reactive azo dye) from aqueous solution. In batch experiments, parameters studied include contact time, adsorbate concentration, pH, adsorbent dose and temperature. Adsorption followed Langmuir isotherm with adsorption capacity of 10.37 and 3.28 mg/g for acid brilliant blue and procion red, respectively. Adsorption kinetic studies showed second order with respect to acid brilliant blue and first order with respect to procion red. Thermodynamic parameters such as free energy, enthalpy and entropy of adsorption were also evaluated. PMID- 17913207 TI - Prompt laboratory diagnosis in timely containment of a plague outbreak in India. AB - A focal outbreak of pneumonic plague occurred in a hamlet of village Hatkoti, district Shimla, Himachal Pradesh in the first fortnight of February, 2002. A total of 16 cases with 4 deaths were reported. Diagnosis of plague was confirmed by the laboratory in 10 (63%) cases. Y. pestis was isolated from clinical samples of 3 cases and confirmed by bacteriophage lysis. Molecular tests confirmed the presence of Y. pestis specific pla and F1 genes in 4 cases; DNA fingerprinting had identity with the known sequence of plague bacilli. Paired samples from 5 cases showed more than 4 fold rise and 1 case showed more than 4 fold fall in antibodies against F1 antigen of Y. pestis. The present communication emphasises that timely and systematic laboratory investigations give confirmatory diagnosis in shortest possible time which forms the backbone of the outbreak control in a timely fashion and prevents confusion and controversy. PMID- 17913208 TI - Detection of M. tuberculosis in clinical samples of diversified nature by IS6110 based PCR. AB - Performance of the polymerase chain reaction technique based on IS6110 sequence was evaluated in clinical samples obtained from pulmonary and extrapulmonary cases of tuberculosis. One hundred and seventy two samples were processed for detection of M. tuberculosis by ZN stained smear examination, LJ medium culture, BACTEC radiometric culture and PCR tests amplifying 123bp region of IS6110 sequence. A significant difference was seen in the sensitivities of different tests, the figures being 83% for PCR test, 35.2% for smear examination, 47.16% for LJ culture and 53.45% for BACTEC culture (p < 0.05). However, no significant difference was found as far as specificity was concerned. PCR test sensitivity in. pulmonary and extrapulmonary clinical samples were 90.14% and 77.27% respectively and found to be significantly higher (p < 0.05) when compared with those of other tests. The mean detection time for M. tuberculosis was 24.03 days by LJ medium culture, 12.89 days by BACTEC culture and less than one day by PCR test. PCR based on IS6100 sequence is highly sensitive method for the early diagnosis of pulmonary and extrapulmonary tuberculosis. PMID- 17913209 TI - Mass drug administration against lymphatic filariasis: experiences from Kozhikode district of Kerala State. AB - The mass DEC drug administration to eliminate lymphatic filariasis in Kozhikode district was monitored from 2001 to 2003 to assess the drug distribution coverage, compliance, reasons for non-compliance, side reactions, mf prevalence and intensity, infection and infectivity rates in the vector. The drug distribution coverage and compliance were much below the required level. "No disease so not necessary" (42.5%) and "fear of side reactions" (25.2%) were the two major reasons for non-compliance. The adverse reactions were minimal. No appreciable changes were found in the mf prevalence and intensity. For the successful implementation of the MDA programme, proper planning, intense and timely efforts to motivate the community and innovative drug delivery strategies are required. PMID- 17913210 TI - Risk factors of herpes simplex virus type 2 among STD clinic attenders in Delhi, India. AB - The present study was conducted to determine the seroprevalence and risk factors associated with HSV-2 infection among sexually transmitted diseases (STD) clinic attenders of Delhi in India. Out of 128 patients included, 76 were males and 52 were females. Antibodies to HSV 1 and 2 and HIV infection were determined by ELISA. Syphilis seropositivity was determined by VDRL test and confirm by TPHA test. Ulcer scrapping were stained by Giemsa for Herpes progenitalis and Donovan bodies and Grams for Haemophilus decreyi infection. The HSV-2 and HSV-I seroprevalence was found to be 85.2% and 77.3% respectively. 87.3% of HSV-2 seropositive patients were asymptomic. 10.7% of patients had coinfection of HSV-2 and HIV. STDs like syphilis, chancroid, gonococcal and non-gonococcal urethritis were significantly associated in HSV-2 infection. Thus the study demonstrates high prevalence of HSV-2 infection in Delhi city. Significant association of HSV 2 infection with previous history of STD (p < 0.02) and multiple sexual partners in males was found (p < 0.002). PMID- 17913211 TI - Perception of risk and potential occupational exposure to HIV/AIDS among medical interns in Delhi. AB - A cross sectional study was conducted among 129 medical interns of Maulana Azad Medical College, New Delhi for assessing the perceived levels of risk of acquiring HIV infection in the health care settings among medical interns, reasons for the same and their exposure to situations having potential of HIV transmission. Majority of the interns (68.3%) perceived themselves to be at a very high/high risk of acquiring HIV infection during their medical career. The common reasons for perceived risk of acquiring HIV infection were getting injuries due to needle pricks/cuts during surgical procedures (32.4%), frequent exposure to the blood/secretions of patients (28.5%) and insufficient availability of gloves (17.6%). Some (23.2%) were of the opinion that students in future might lose interest in the medical profession due to increasing risk of HIV infection and few (3.1%) were even considering to leave the medical profession for the same reason. Majority of the interns (72.9%) had experienced needle pricks and more than half (53.7%) of them even had had blood splashes in their eyes/nose/mouth during surgical procedures. The findings of the study call for efforts for bringing a reduction in the risk perception of the interns through awareness campaigns and reorientation trainings, ensuring availability of gloves and other items necessary for observing universal work precautions and proper disposal of potentially contaminated articles. PMID- 17913212 TI - Trichrome staining for detection of intestinal protozoa a better screening method. AB - Intestinal protozoal infections are common in our country because of poor hygiene and tropical conditions. The efficacy of trichrome staining to screen stool smear was compared with commonly used methods i.e. concentrated iodine mount and direct wet mount to test its better effectiveness. All Stool samples were first examined by routine methods i.e. direct wet mount and iodine staining. A portion of stool sample was also inoculated in vial containing polyvinyl alcohol (PVA) fixative. From PVA preserved samples, slides were prepared and stained by modified wheately's trichrome method. The results of both methods were compared and relative accuracy was calculated. 1054 stool specimens were examined and 259 parasites detected, of which 20.7% were protozoa and 3.7% helminthde. Trichrome staining detected 19.1% protozoa while routine methods detected 12.9% protozoa. For identification of protozoa, accuracy was 91.8% in favor trichrome staining and 61.8% by wet mount and iodine staining. Trichrome stained smear alone can be used as screening method in those geographic areas where protozoa infections are common. PMID- 17913213 TI - Knowledge, attitude and practices about animal bites and rabies in general community--a multi-centric study. AB - Rabies a disease as old as our civilization, continues to be the most feared of all communicable diseases. Despite the availability the state-of-the-art tools which ensure near cent percent protection against rabies, India is the largest contributant to rabies mortality in the world. A multicentric study was carried out from April 2001 to September 2002 with the objective of assessing the knowledge, attitudes, beliefs and practices (KAP) about animal bites and rabies in the general community. The proforma for interviewing the general community was developed and used after field testing. The study was carried out at six selected centres across the country viz. Delhi, Hyderabad, Raipur, Jamnagar, Coonoor and Rajahmundry and was co-ordinated by National Institute of Communicable Diseases (NICD), Delhi, after thorough briefing of designated nodal officers. A total of 1129 (male: female :: 48.5: 51.5) persons in the age group of 18 to 80 years were interviewed in this study. Of these about 751% of the individuals had attended school at some level and rest were illiterates. 68.7% people had heard about rabies. In 60.7% of cases the community associates rabies with dog bite only. Knowledge about appropriate wound toilet was found to be inadequate. Only 360 (31.9%0/) people felt that washing the wound with soap and water was the best option. Application of indigenous products like chillies (11.4%), turmeric (5.6%), lime (6.8%), kerosene oil (2.3%), herbal paste (4.2%) etc was suggested along with visit to occult medicine practitioner (1.5%) as part of the bite wound management. People were not aware of number of injections needed for treatment of animal bites. Multiple reasons viz negligence and ignorance 354 (31.4%), fear of multiple painful injections 365 (32.3%), expensive treatment 169 (15%) and long course requiring daily visits to anti-rabies clinics 73 (6.5%) were cited as reasons for non-compliance of treatment. KAP study suggests that there is need to create awareness amongst the masses regarding epidemiology of the disease and merits of prompt and appropriate post exposure treatment through enhanced IEC activities. PMID- 17913214 TI - Profile of bacterial isolates from suspected cases of infective endocarditis (IE): a retrospective study in a north Indian tertiary care hospital. AB - Infective Endocarditis (IE) is an emerging infection of the twenty-first century. This chronic Infection is mainly caused by bacteria, although fungi can also be associated with it. It is Important to know the profile of bacteria causing IE in a given region so as to suggest the empirical therapy for this serious illness. Blood culture isolates of clinically diagnosed or suspected cases of IE admitted to various wards of the All India Institute of Medical Sciences were analyzed retrospectively from January 2000 to June 2004. Standard techniques were used for the isolation and identification of the bacteria. Our study has demonstrated the predominance of Gram-negative bacilli, especially, Acinetobacter species and Pseudomonas aeruginosa, which are notorious for antimicrobial resistance, as the aetiological agents of IE. Amongst Gram-positive cocci, Enterococci exhibiting HLAR comprised the predominant species. Methicillin resistance among staphylococcal strains in this Tertiary care hospital is adding to the therapeutic challenge in the management of this serious illness. Although antimicrobial treatment should not be delayed in such cases, we cannot undermine the importance of isolation and identification of the etiological agents and the determination of the antimicrobial susceptibility for the management of these life-threatening conditions as well as for the formulation of guidelines for empirical therapy of these cases. PMID- 17913216 TI - Post tsunami--a survey of mosquito fauna and breeding habitats in Teressa Island, Andaman and Nicobar Island, India. PMID- 17913215 TI - A note on the susceptibility status of An. culicifacies and An. fluviatilis to malathion in Nainital and Udham Singh Nagar districts, Uttarakhand. PMID- 17913217 TI - Influence of larval diet on the autogenous egg production in Phlebotomus papatasi (Diptera: Phlebotomidae) the vector of leishmanisis. PMID- 17913218 TI - Antibiotic resistance pattern in Acinetobacter spp.--a retrospective anaysis. PMID- 17913219 TI - Bacteriological quality of potable water in Davangere City. PMID- 17913220 TI - Relevance of the Widal test today. PMID- 17913221 TI - Risk assessment of dietary supplements. AB - Risk assessment of dietary supplements shares many of the requirements of that for other chemicals, although there are some important differences. Amongst these is the essential nature of some nutrients so that it may be necessary to balance the need to minimize toxicological risk with the need to avoid deficiency. There may also be limitations on experimental design, in that high doses may not be achievable for nutritional reasons and available human data on toxicological hazard is likely to be very limited. Prior to embarking on a risk assessment the problem needs to be formulated. This involves risk assessors, risk managers and relevant stakeholders. A key decision is whether a risk assessment is necessary and, if so, what is required of the assessment. This will shape the nature and output of the assessment. Risk assessment itself is a scientific process comprising four steps, hazard identification, hazard characterization, exposure assessment and risk characterization. Hazard identification involves determining the range of toxicological effects that might be caused by the substance, whilst hazard characterization establishes dose-response relationships, toxicological and species relevance of the findings and establishes health based guidance values. Exposure assessment involves predicting or measuring the level, pattern and duration of intake of the substance by exposed individuals. This may require dietary consumption data. Finally, risk characterization is the process whereby all of the prior information is integrated to reach conclusions in a form appropriate to the question posed. The nature of the output can take several different forms, and may be qualitative or quantitative. There are some cross cutting issues in risk assessment, primarily on uncertainty and variability. The sources of uncertainty at each step of the risk assessment should be clearly identified and quantified to the extent possible. Variability requires that the risk assessment should take into account all relevant subpopulations and groups, where there might be differences on the basis of intake or sensitivity. This would very often include different life stages, but may also include gender, ethnic or genetic differences. The report of the risk assessment should be systematic and transparent, identifying all key assumptions and defaults used. PMID- 17913222 TI - Setting dietary intake levels: problems and pitfalls. AB - Recommended dietary intake levels are the nutrient standards used in designing food assistance programmes, institutional feeding programmes, counselling and teaching. In the USA, the recommended dietary allowances (RDAs) are the basis for setting the poverty threshold and food stamp allotments. In the 1990s, a new paradigm was put forth for estimating nutrient requirements and recommended intake levels. This considered the level of nutrient needed for normal body functioning (versus the amount needed to prevent a deficiency state from occurring). An estimated average requirement (EAR), an RDA and a tolerable upper intake level (UL) were determined for most nutrients. In setting forth these nutrient intake levels (dietary reference intakes, DRIs), a number of data challenges were encountered. For example, it was recognized that for most nutrients there was an absence of dose-response data, and few chronic human or animal studies had been undertaken. In considering how to revise nutrient intake recommendations for populations in the future, the following pitfalls must be overcome: (1) invalid assumption that a threshold level for a requirement will hold for all nutrients; (2) lack of uniform criteria for the selection of the endpoints used (need for evidence-based review, consideration of comparative risk); (3) invalid extrapolations to children for many nutrients; (4) lack of information on variability of responses, and interactions with other nutrients; and (5) lack of understanding in the community of how to use the various DRI numbers. PMID- 17913223 TI - Criteria for substantiating claims. AB - Claims are used to support public health advocacy and marketing. Their evidence base is variable. Claims are made on (i) nutrient content, (ii) comparative merits, (iii) health benefits, and (iv) medical benefits. Experience with therapeutic agents has aided the development of recommendations for the substantiation of health claims for foods and food components, with which dietary supplements would be included. An EU Concerted Activity, Functional Food Science in Europe, suggested that such claims should be based on the general outcomes of 'enhanced function' and 'reduced risk of disease'. A further EU Concerted Activity, The Process for the Assessment of Scientific Support for Claims on Foods, proposed that the evidence base should provide: a characterization of the food or food component to which the claimed effect is attributed; human data, primarily from intervention studies that represent the target populations for the claim; a dose-response relationship: evidence of allowing for confounders including lifestyle, consumption patterns, background diet and food matrix; an appropriate duration for the study; a measure of compliance; and have adequate statistical power to test the hypothesis. When ideal endpoints are not easily accessible for measurement, validated and quality assured markers of the intermediate or final outcomes could be used, as long as their relationship is well characterized. Overall, the totality and coherence of published and unpublished evidence should be considered. Assessments for substantiation need expert judgement, weighting of the strength of the claim, and intelligent use of the criteria applied on an individual basis with respect both to gaps in knowledge and to any need for new knowledge and data. PMID- 17913224 TI - Science in the regulatory setting: a challenging but incompatible mix? AB - Regulatory decisions informed by sound science have an important role in many regulatory applications involving drugs and foods, including applications related to dietary supplements. However, science is only one of many factors that must be taken into account in the regulatory decision-making process. In many cases, the scientific input to a regulatory decision must compete with other factors (e.g. economics, legal requirements, stakeholder interests) for impact on the resultant policy decision. Therefore, timely and effective articulation of the available science to support a regulatory decision can significantly affect the relative weight given to science. However, the incorporation of science into the regulatory process for dietary supplements is often fraught with challenges. The available scientific evidence has rarely been designed for the purpose of addressing regulatory questions and is often preliminary and of widely varying scientific quality. To add to the confusion, the same scientific evidence may result in what appears to be different regulatory decisions because the context in which the science is used differs. The underlying assumption is that scientists who have a basic understanding of the interface between science and policy decisions can more effectively provide scientific input into these decisions. PMID- 17913225 TI - Vitamin E. AB - Vitamin E refers to a family of tocopherol and tocotrienol isomers discovered in 1922 as anti-infertility factor. Vitamin E deficiency causes infertility and delayed-onset ataxia in experimental animals, and it leads to neuronal dysfunctions in humans. However, based largely on its radical-scavenging antioxidant activity in vitro, vitamin E supplements are commonly thought to provide health benefits against diseases associated with oxidative damage, most notably cardiovascular diseases. Contrary to this belief, the outcome of recent large, prospective, randomized and placebo-controlled clinical studies does not encourage the use of vitamin E supplements. These overall disappointing results can be explained and substantiated by scientific data critically testing the strengths of evidence for many of the underlying assumptions and examining the possibility that in vivo vitamin E may have function(s) other than, or in addition to, acting as an antioxidant. PMID- 17913226 TI - Flavonoids: a re-run of the carotenoids story? AB - Flavonoids have powerful antioxidant activities in vitro, but the evidence that they act as antioxidants in vivo in humans is equivocal at best. However, they may be able to help protect the gastro-intestinal tract against reactive oxygen species. PMID- 17913227 TI - Reduced folate status is common and increases disease risk. It can be corrected by daily ingestion of supplements or fortification. AB - The natural folates are chemically unstable and poorly bioavailable in contrast to the chemical form, folic acid. Consequently most people, even those on good diets, have less than optimal nutrition with respect to this vitamin. Increased risks associated with deficiency include neural tube defects (NTDs) (proven), ischaemic heart disease and stroke (probable), certain cancers and decline in cognitive function (possible). Supplements of folic acid at the population reference intake (400 microg/d) completely normalizes all of these risks. Such levels are safe as judged by decades of use in wide sectors of the population. The main drawback of supplements is with respect to their effectiveness in preventing NTDs. They must be taken periconceptionally and before most women realise that they are pregnant. No more than one fifth of women take supplements effectively, largely due to the fact that over half of pregnancies are unplanned. This has led to the alternative of mandatory fortification of flour with folic acid in the USA and Canada. The level for such fortification is suboptimal for NTD prevention, because of fear of overexposure in the elderly. Thus even fortified communities require advice to take supplements for optimum NTD prevention. PMID- 17913228 TI - Calcium and vitamin D. AB - Calcium is required for normal growth and development as well as maintenance of the skeleton. Vitamin D is also essential for intestinal calcium absorption and plays a central role in maintaining calcium homeostasis and skeletal integrity. In addition, both micronutrients have important roles in non-skeletal-related physiological processes. Of concern, significant proportions of some population groups fail to achieve the recommended calcium intakes in a number of western countries. Furthermore, while cutaneous biosynthesis upon exposure of skin to ultraviolet B light is the major source of vitamin D for most people, this does not occur during winter time. Thus, there is an increased reliance on dietary sources during winter months to help maintain adequate vitamin D status. Since vitamin D is found naturally only in a limited number of foods, the usual dietary vitamin D intake by many European populations is not sufficient to maintain adequate vitamin D status. This paper will briefly review these important issues together with consideration of the potential role for supplementation with calcium and vitamin D in terms of improving their intakes in the population. It will also focus on the issues of efficacy as well as safety considerations of supplements. PMID- 17913230 TI - Herbal medicines: balancing benefits and risks. AB - Herbal medicines are preparations containing exclusively plant material. Their efficacy can be tested in clinical trials much like synthetic drugs but numerous methodological and logistical problems exist. For several herbal medicines, efficacy has been established; for many others, this is not the case mostly because the research has not been done. Many consumers believe that herbal medicines are natural and therefore safe. This is a dangerous simplification. Some herbal medicines are associated with toxicity others interact with synthetic drugs. The often under-regulated quality of herbal medicines amounts to another safety issue. Contamination or adulteration of herbal medicines are possible and can cause harm. In order to conduct a risk-benefit analysis of a specific herbal medicine for a specific indication, we require definitive efficacy and safety data. This is currently the case for only very few such preparations. It follows that, in order to advise consumers responsibly, the gaps in our present knowledge require filling. PMID- 17913229 TI - Selenium. AB - Selenium occurs as inorganic selenite or selenate and in organic forms in plants and other organisms used for food. The human selenoproteome consists of 25 selenoproteins. The main groups are glutathione peroxidases 1-5, iodothyronine deiodinases 1-3, thioredoxin reductases, selenoprotein P (SelP), and other proteins mostly with unknown function. In selenoproteins selenium occurs as selenocysteine. SelP works as a transporter of selenium between the liver and other organs. Selenium in the form of selenomethionine can also unspecifically substitute for methionine in other proteins. No specific deficiency condition has been described in humans. The aetiology of Keshan disease, a cardiomyopathy, is a combination of coxsackie virus and low selenium. Selenium status has been linked to the incidence of cancer and other diseases. Excess selenium can produce selenosis in humans affecting liver, skin, nails and hair. Recommended intake and upper tolerable level are 40-55 and 300 microg/day. A better chemical characterization of selenium compounds in foods and in particular supplements as well as knowledge on the apparent differences in biological activity between selenium compounds, both with respect to nutrition, disease protection and adverse effects, are needed. Supplementation studies should in addition to possible beneficial effects also focus on the possibility of possible adverse effects. PMID- 17913231 TI - Standardization and evaluation of botanical mixtures: lessons from a traditional Chinese herb, Epimedium, with oestrogenic properties. AB - Botanical extracts differ from conventional supplements in that they are complicated mixtures of many bioactive compounds. Here we describe our experience with a traditional Chinese medicinal plant Epimedium sp. to illustrate the scientific challenges of firstly, obtaining a standardized product from a complex mixture and secondly, evaluating that product for preclinical and clinical efficacy. In contrast, to its colloquial name 'Horny goat weed' and Internet advertisements as a herbal 'Viagra' for men, extracts of Epimedium are strongly oestrogenic due to the presence of novel potent phytoestrogens of the prenyl flavone family. Since Epimedium is not cultivated, it was necessary to source for taxonomically identified samples and to authenticate their species by phylogenetic, chemical and bioresponse profiling. The feasibility of using a panel of oestrogen-responsive cell-based bioassays to measure summated oestrogenic effects at close time points for pharmacokinetic/pharmacodynamic (PK/PD) modelling was evaluated. We document proportionate oestrogenic responses in sera of animals fed oestrogenic drugs and botanical extracts, indicating that these target molecule responsive cell-based bioassays may have utility to capture the global effects of the myriad bioactive compounds in botanical extracts, informing the design of rigorous clinical trials for safety and efficacy. PMID- 17913232 TI - Communication between science and management. AB - Four methods of communication between science and management are conceivable: (1) authoritative science, which provides results without explanation or justification; (2) authoritative management with imposition of a preconceived management decision on science, which destroys independence and credibility of the scientist; (3) usurpation or mutual invasion of both science and management into each other's territory, which is detrimental to the integrity of both; and (4) interaction between scientist and manager, in which the different tasks of both are recognized and respected. For the latter it is important to accept that the commitment of the scientist is to science only and that managers are committed to other considerations besides science. The role of the scientist is easier because of their commitment, and the results of their work are less likely to be questioned, when they succeed in communicating their working methods, treatment of (missing) data, deductions, and results in a comprehensible and logical form. The manager, however, will be in a difficult position, if these results take the form of advice which, for whatever reasons, they cannot follow. Managers may be tempted to avoid advice or, if unavoidable, to doubt its correctness or to modify its meaning, instead of justifying their decision. The major problem in communication between science and management is probably in semantics: the wording of the task of the scientist and of the scientific result in unambiguous language which is understood by the assessor and the manager, respectively. PMID- 17913233 TI - Dietary supplements and health: the research agenda. AB - Research needs to evaluate the role of dietary supplements in human health abound, yet funds to support all of the possible opportunities do not. Government agencies, such as the National Institutes of Health (NIH) in the USA, remain the chief sponsors of research in this area. They face the challenge of competing priorities, such as critical disease-oriented research, basic biomedical and technological development, and prevention-related research. Dietary supplements are widely used for health promotion and disease prevention, sometimes with minimal science to support their use. There is a need for focused research efforts to better address issues of efficacy, safety and quality of dietary supplements. At the same time, fundamental studies of their mechanisms of action are needed. In addition, resources to support research in this area are required: on the one hand, basic tools (analytical methods, characterization of ingredients) need to be developed and validated, and on the other, tools to understand patterns of supplement use in populations and study designs to assess their efficacy and safety need refining. These efforts benefit greatly from partnerships among government agencies and with the academic and private sectors. PMID- 17913234 TI - Fatty acid composition of triacylglycerols from Wharton's jelly determined by high-performance liquid chromatography. AB - The improved method for HPLC determination of fatty acids was proposed. The chromatographic separation of p-bromophenacyl derivatives of fatty acids under a gradient elution was achieved at 40 degrees C with an RP-18 LiChroCART 5 column and organic mobile phase containing methanol, acetonitrile, water and TEAP buffer pH 5.6. The quantitative determination of those derivatives was performed at 254 nm. Preeclampsia, the most common pregnancy complication, did not affect triacylglycerol content in the umbilical cord Wharton's jelly in comparison to the control material. However, it changed the composition of fatty acids, bound to that lipid class. The method allows the determination of almost all fatty acids forming the investigated neutral lipid class, contained in a solid tissue sample. The use of TEAP buffer excluded precipitation and flow stoppage in the HPLC system. The method reduced time and costs and might be useful for all other lipid classes and different tissues. PMID- 17913235 TI - Analytical approach for selecting normalizing genes from a cDNA microarray platform to be used in q-RT-PCR assays: a cnidarian case study. AB - Research in gene function using Quantitative Reverse Transcription PCR (q-RT-PCR) and microarray approaches are emerging and just about to explode in the field of coral and cnidarian biology. These approaches are showing the great potential to significantly advance our understanding of how corals respond to abiotic and biotic stresses, and how host cnidarians/dinoflagellates symbioses are maintained and regulated. With these genomic advances, however, new analytical challenges are also emerging, such as the normalization of gene expression data derived from q-RT-PCR. In this study, an effective analytical method is introduced to identify candidate housekeeping genes (HKG) from a sea anemone (Anthopleura elegantissima) cDNA microarray platform that can be used as internal control genes to normalize q-RT-PCR gene expression data. It is shown that the identified HKGs were stable among the experimental conditions tested in this study. The three most stables genes identified, in term of gene expression, were beta-actin, ribosomal protein L12, and a Poly(a) binding protein. The applications of these HKGs in other cnidarian systems are further discussed. PMID- 17913236 TI - A method for investigating the temporal dynamics of local neuroretinal responses. AB - Visual sensitivity improves with prolonged exposure to light. Global neuroretinal responses increase, but little is known about the dynamics of local retinal responses over brief time intervals after changes in light level. This study applies the time-slice multifocal electroretinogram (TS mfERG) paradigm for the measurement of local electrical responses of the human eye over brief time intervals. Sixty-one, localised retinal areas were assessed over 25 degrees of the visual field. Cone-mediated contributions to the time-slice waveform were established. The time-slice mfERG waveforms were similar in shape and timing for pre- and post-photopigment bleach conditions after saturation of rod-mediated responses, suggesting there was no rod-mediated intrusion in the waveform. The temporal dynamics of the mfERG components show that N1P1 amplitudes decrease with each successive time-slice probe, with larger amplitude responses in the central retina compared to nasal and temporal retina. The time-slice mfERG waveform is a technique for assessing the temporal dynamics of cone-generated neural responses over time. The data are interpreted in terms of the vascular supplies and lower level visual adaptation mechanisms. PMID- 17913237 TI - A novel group analysis for functional MRI of the human brain based on a two threshold correlation (TTC) method. AB - This work presents a new group analysis for functional MRI of human brain activation. The two-threshold correlation (TTC) method determines two statistical thresholds by estimating the noise distribution underlying the summed histogram of correlation coefficients (CC) from all sections and subjects. The probabilistic CC thresholds (p<0.0001 for the identification of highly significant activation centers and p<0.05 for limiting the iterative addition of directly neighboring voxels to these centers) are applied to the group CC maps for each section. These maps may be reconstructed by taking the maximum (MAX) or mean (MEAN) CC value of all subjects for a particular voxel. Experimental analyses involved functional echo-planar imaging of sequential finger-to-thumb opposition and silent word generation at 3T (eight subjects). Preprocessing included motion correction, spatial filtering, and normalization to MNI space. While the results for the TTC MAX approach were very similar to those obtained for a standard SPM analysis, the TTC MEAN approach turned out to be more conservative emphasizing voxels that are activated in most rather than in only a few subjects. The new method is simple, fast, and robust by linking two thresholds in a physiologically meaningful manner. PMID- 17913238 TI - Mental healthcare status and psychiatric diagnoses of train suicides. AB - BACKGROUND: The objective of this study is to investigate mental healthcare status and psychiatric disorders in train suicides. METHODS: Data of 4 published train suicide studies were combined with a study of 57 train suicides in The Netherlands. RESULTS: 53% of all train suicides received psychiatric care at the time of suicide, with 49% of them being inpatients. These values are higher than those found in general suicides. When compared to general suicides, functional non-affective psychoses are overrepresented by 25% vs 14%. The percentage of affective disorders approximates that of general suicides namely: 39% vs 42%. "Other diagnoses" are underrepresented by 23% vs 40%. LIMITATIONS: All psychiatric diagnoses were based on clinical data, which may not accurately reflect the patient's psychiatric condition at the time of suicide. CONCLUSIONS: Train suicides receive mental healthcare more often than general suicides and are more often characterized by severe psychopathology. The study indicates that patients with affective and psychotic disorders in particular should be targeted in order to prevent train suicide. PMID- 17913239 TI - Ex vivo cytokine production by whole mid-trimester amniotic fluid. AB - We hypothesized that ex vivo measurement of intraamniotic production of immune mediators differed from analysis of these mediators within unincubated amniotic fluid. Mid-trimester amniotic fluid from 72 women were incubated ex vivo with or without 50 ng/ml lipopolysaccharide (LPS). Supernatants and the corresponding unincubated amniotic fluids were tested for interleukin (IL)-6, IL-1 receptor antagonist (IL-1ra), IL-10 and nitric oxide. Ex vivo culture resulted in increased release of IL-6, IL-10 and nitric oxide; IL-1ra levels were decreased following the incubation. A spontaneous preterm birth (SPTB) occurred in 12 (16.7%) of the subjects. Women with a subsequent SPTB had decreased IL-6 and increased IL-10 production following ex vivo culture compared to women with a term delivery. This association was not evident with unincubated amniotic fluids. Conversely, IL-1ra concentrations were elevated in women with subsequent SPTB only in unincubated amniotic fluids. Immune mediator production by ex vivo amniotic fluid culture differs from that present in amniotic fluid supernatants and may provide a more accurate indication of the immune potential of the intraamniotic environment. PMID- 17913240 TI - An experimental pilot study of response to invalidation in young women with features of borderline personality disorder. AB - One of the leading biosocial theories of borderline personality disorder (BPD) suggests that individuals with BPD have biologically based abnormalities in emotion regulation contributing to more intense and rapid responses to emotional stimuli, in particular, invalidation [Linehan, M.M., 1993. Cognitive-Behavioral Treatment of Borderline Personality Disorder. Guilford, New York.]. This study used a 2 by 2 experimental design to test whether young women with features of BPD actually show increased physiological arousal in response to invalidation. Twenty-three women ages 18 to 29 who endorsed high levels of BPD symptoms and 18 healthy controls were randomly assigned to hear either a validating or invalidating comment during a frustrating task. Although we found preliminary support for differential response to these stimuli in self-report of valence, we found neither self-report nor physiological evidence of hyperarousal in the BPD features group, either at baseline or in response to invalidation. Interestingly, the BPD features group reported significantly lower comfort with emotion, and comfort was significantly associated with affective valence but not arousal. We discuss implications for understanding and responding to the affective intensity of this population. PMID- 17913241 TI - Temporal patterns of self-injurious behavior correlate with stress hormone levels in the developmentally disabled. AB - While the origins and developmental course of self-injurious behavior (SIB) remain relatively unknown, recent studies suggest a biological imbalance may potentiate or provoke the contagious recurrence of SIB patterns in individuals with severe developmental disabilities (DD). Evidence from several laboratories indicates that functioning, relations, and processing of a stress-related molecule, proopiomelanocortin (POMC) may be perturbed among certain subgroups of individuals exhibiting SIB. The current investigation employed a unique time pattern analysis program (THEME) to examine whether recurrent temporal patterns (T-patterns) of SIB were related to morning levels of two POMC-derived hormones: beta-endorphin (betaE) and adrenocorticotropic hormone (ACTH). THEME was used to quantify highly significant (non-random) T-patterns that included SIB within a dataset of in situ observational recordings spanning 8 days ( approximately 40 h) in 25 subjects with DD. Pearson's product-moment analyses revealed highly significant correlations between the percentage of T-patterns containing SIB and basal levels of both betaE and ACTH, which were not found with any other "control" T-patterns. These findings support the hypothesis that the recurrent temporal patterning of SIB represents a unique behavioral phenotype directly related to perturbed levels of POMC-derived stress hormones in certain individuals with severe DD. PMID- 17913242 TI - Synergistic effects of IL-2, IL-12 and IL-18 on cytolytic activity, perforin expression and IFN-gamma production of porcine natural killer cells. AB - Natural killer (NK) cells are one of the main cellular components of the innate immune system. They play an important role in the immune response against infections as well as tumour cells and therefore have two major properties: production of immune regulatory cytokines and chemokines as well as cytolytic destruction of particular target cells. The existence of NK cells in swine is well known as well as the phenotype of resting NK cells, but their response following activation by cytokines is still poorly understood. Therefore, we tested the influence of the immune regulatory cytokines IL-2, IL-12 and IL-18 on cytolytic activity, phenotype, IFN-gamma production and the accumulation of perforin in cytoplasm of peripheral blood mononuclear cells (PBMC) as well as purified NK cells. NK cells were enriched from PBMC using a magnetic cell separation (MACS) strategy with monoclonal antibodies against CD3, CD21 and SWC3, thereby removing T-, B- and myeloid cells. Respective fractions were used in flow cytometry (FCM) based cytolytic assays with the human tumour cell line K562 as target. After stimulation with the cytokines described above, the NK cell enriched CD3(-)CD21(-)SWC3(-) fraction showed an evident increase in the cytolytic activity compared to PBMC. This enhanced cytolytic activity was accompanied by a strong enrichment of IFN-gamma producing cells when a combination of all three cytokines (IL-2/IL-12/IL-18) was used; as determined in ELISPOT assays and intracellular staining of IFN-gamma in FCM. Also, the combination of these three cytokines led to an accumulation of perforin in the cytoplasm and an up-regulation of CD25 compared to control cultures incubated in medium without cytokines. The experiments performed clearly indicate a stimulatory role and strong synergistic effects of the investigated cytokines in the activation of porcine NK cells in vitro, inducing IFN-gamma, perforin production and cytotoxicity against target cells. PMID- 17913243 TI - Adenosine A2A receptor agonists inhibit lipopolysaccharide-induced production of tumor necrosis factor-alpha by equine monocytes. AB - Adenosine is an endogenous nucleoside that regulates many physiological processes by activating one or more adenosine receptor subtypes, namely A1, A2A, A2B and A3. The results of previous studies indicate that adenosine analogues inhibit lipopolysaccharide (LPS)-induced production of reactive oxygen species (ROS) by equine neutrophils primarily through activation of A2A receptors. Because peripheral blood monocytes produce cytokines that are responsible for many of the deleterious effects of LPS, the current study was performed to evaluate the effects of an array of novel adenosine receptor agonists on LPS-induced production of tumor necrosis factor-alpha (TNF-alpha), and to assess the selectively of these agonists for equine adenosine A2A over the A1 receptor. Radioligand binding studies performed with equine tissues expressing adenosine A1 and A2A receptor subtypes yielded a rank order of affinity for the equine A2A receptor of ATL307>ATL309 approximately ATL310 approximately ATL313>ATL202 approximately ATL361 approximately ATL376>ATL372>CGS21680>NECA. Co-incubation of equine peripheral blood monocytes with LPS and these agonists resulted in inhibition of TNF-alpha production with a rank order of potency that strongly correlated with their binding affinities for equine adenosine A2A receptors. Results of experiments performed with one of the adenosine receptor agonists (ATL313) and selective adenosine receptor antagonists confirmed that inhibition of LPS-induced production of TNF-alpha occurred via stimulation of A2A receptors. Although incubation of monocytes with IB-MECA, a compound purported to act as an adenosine A3 receptor agonist, reduced LPS-induced TNF-alpha production, this effect of IB-MECA was inhibited by the A2A selective antagonist ZM241385 but not by the A3 receptor antagonist MRS1220. These results indicate that the adenosine receptor subtype responsible for regulation of LPS-induced cytokine production by equine monocytes is the A2A receptor. To address the signal transduction mechanism responsible for the anti-inflammatory effects of ATL313 in equine monocytes, production of cAMP was compared in the presence and absence of either the adenosine A2A receptor antagonist ZM241385 or the adenosine A2B receptor antagonist MRS1706. In the absence of the antagonists, ATL313 increased production of cAMP; ZM241385 inhibited this effect of ATL313, whereas MRS1706 did not. Furthermore, incubation of monocytes with either the stable analogue of cAMP, dibutyryl cAMP, or forskolin, an activator of adenylyl cyclase, also inhibited LPS-induced production of TNF-alpha production by equine monocytes. Collectively, the results of the current study indicate that adenosine analogues inhibit LPS-induced production of TNF-alpha by equine monocytes primarily via activation of adenosine A2A receptors and do so in a cAMP-dependent manner. The results of this study indicate that stable adenosine analogues that are selective for adenosine A2A receptors may be suitable for development as anti-inflammatory drugs in horses. PMID- 17913244 TI - Cloning of canine Toll-like receptor 7 gene and its expression in dog tissues. AB - Toll-like receptor 7 (TLR7) is activated by single strand RNA and imidazoquinoline compounds, and induces interferon production. In this study, canine TLR7 cDNA was cloned and sequenced. The full-length cDNA of canine TLR7 gene was 3419bp, encoding 1032 amino acids. The similarities of canine TLR7 with human and mouse TLR7 were 84 and 80% at the nucleotide sequence level, and 86 and 79% at amino acid sequence level, respectively. Further, the expression of TLR7 mRNA was investigated in canine normal tissues by semiquantitative RT-PCR analysis. The common expression level of TLR7 mRNA in tissues from three dogs examined was in large intestine, lung, pancreas, small intestine and skin, though the expression level in each tissue was varied among these healthy dogs. In other tissues (kidney, liver, lymph node, spleen, adrenal gland, and PBMCs), the level of TLR7 mRNA expression was different in individuals. PMID- 17913245 TI - Synergistic antitumor effect of chemotactic-prostate tumor-associated antigen gene-modified tumor cell vaccine and anti-CTLA-4 mAb in murine tumor model. AB - In this study, we demonstrate that an effective immune response against prostate tumors in mouse tumor model can be elicited using a strategy that combines CTLA-4 blockade and pSLC-3P-Fc-modified tumor cell vaccine (named B16F10-SLC-3P-Fc). Treatment of B16F10-3P-bearing mice resulted in a significant reduction in tumor incidence as assessed 2 months after treatment. In vivo Ab depletion confirmed that the antitumor effect was primarily CD8+ T cells and CD4+ T lymphocytes were required for the induction of CD8+ CTL response in B16F10-SLC-3P-Fc+anti-CTLA-4 mAb-immunized mice. Moreover, mice that were cured of an established tumor were protected against a rechallenge with the same tumor for at least 4 months, suggesting the generation of memory responses. Adoptive transfer experiments further indicate that antitumor reactivity can be transferred to naive mice by splenocytes. These findings demonstrate that this combinatorial treatment can elicit a potent anti-tumor immune response and suggest potential of this approach for treatment of prostate cancer. PMID- 17913246 TI - Genetic association analysis of the interleukin 7 gene (IL7) in multiple sclerosis. AB - IL7 is a nonredundant cytokine, essential for T cell survival and development in humans. We genotyped nine tagging single nucleotide polymorphisms (SNP), representing all parts of the IL7 gene, in 1,210 Swedish multiple sclerosis (MS) patients and 1,234 healthy controls. None of the SNPs showed a significantly different distribution in MS, and haplotype analysis also failed to reveal differences between patients and controls. We conclude that the IL7 gene is very unlikely to influence the genetic susceptibility to MS in this population. PMID- 17913247 TI - Protective effect of pulmonary surfactant on cilia of Eustachian tube in otitis media with effusion. AB - OBJECTIVE: The aim of this study was to evaluate the effect of pulmonary surfactant (PS) on cilia of Eustachian tube (ET) in guinea pigs suffering from otitis media with effusion (OME). METHODS: A nonviable heat-killed pneumococci (HKP) solution was injected into the tympanum of guinea pig via a transtympanic approach therefore establishing models of OME (effusion in tympanum and hearing loss). After being injected with physiologic saline solution or pulmonary surfactant transtympanically, the response threshold of each guinea pig was taken by ABR. Changes of effusion and ciliary ultrastructure were observed. RESULTS: Normal group (A), the response threshold is 14.00+/-3.08 dBHL. Abundant and intact cilia were observed, ciliary morphology was normal, cilia regularly arranged in concord, no goblet cells was observed. Model-control group (B0), 5 days following the inoculation of HKP, serous effusion was present in tympanum. The response threshold is 45.00+/-5.67 dBHL. Cilia lost and arranged irregularly, hyperplasia of the goblet cells was observed. Saline group, the previously observed serous effusion of tympanum was increased or unchanged. The response threshold is 65.50+/-6.85 dBHL. Cilia were coalescing and swelling. A lot of cilia lost and arranged disorderly. A number of compound cilia of the bulging type were formed by cytoplasmic protuberances of ciliated cells. Massive mucoid secretions stuck on the surface of the ET. PS group, the previously observed serous effusion of tympanum was reduced in amount or disappeared completely. The response threshold is 23.53+/-6.32 dBHL. Ciliary morphology was generally normal. Cilia comparatively arrange in concord. A mucus deposits was adhering to the tips of cilia. CONCLUSION: The findings of the study suggest that pulmonary surfactant plays an important role in protecting of cilia of ET of guinea pigs suffering from OME. PMID- 17913248 TI - Cystic lymphangioma: trauma may be a significant cause? AB - Cystic lymphangioma is a rare congenital anomaly of the lymphatic system. Its embryopathogenesis is still controversial. Most lesions present in infancy or early childhood with a swelling found in the head and neck region. We report the case of a woman who presented with a cystic swelling in the left supraclavicular region that appeared after left side thyroidectomy. Surgical excision achieved complete removal. Histology showed the lesion to be a cystic lymphangioma. It can be hypothesized that an embryogenetic origin of the adult lymphangioma is associated with some precipitating trauma such as surgery. PMID- 17913249 TI - Validation of two commercial real-time RT-PCR kits for rapid and specific diagnosis of classical swine fever virus. AB - Two real-time RT-PCR kits, developed by LSI (TaqVet CSF) and ADIAGENE (Adiavet CSF), obtained an agreement to be commercialised in France, subject to conditions, defined by the French Classical Swine Fever (CSF) National Reference Laboratory. The producers were asked to introduce an internal control to check the RNA extraction efficacy. The different criteria assessed were sensitivity, "pestivirus specificity", reproducibility and ease of handling, using 189 different samples. These samples were either CSFV inactivated strains or blood/serum/organs collected from CSFV experimentally infected pigs or naturally infected wild boars. The reproducibility of the assays was confirmed by the analysis of a batch-to-batch panel control that was used for inter-laboratory tests involving nine laboratories. The two kits were also tested for the use in mass diagnostics and the results proved the kits to be suited using pools of blood, serum and tonsils. Moreover, a field evaluation, carried out on spleen samples collected from the CSF surveillance of wild boars in an area known to be infected and from domestic pigs at a slaughterhouse, confirmed the high sensitivity and specificity of the two kits. This step-by-step evaluation procedure confirmed that the two commercial CSF real-time RT-PCR kits have a higher predictive value than the current diagnostic standard, Virus Isolation. PMID- 17913250 TI - Purification of the major envelop protein GP5 of porcine reproductive and respiratory syndrome virus (PRRSV) from native virions. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) is the cause of an economically important swine disease that has been devastating the global swine industry since the early 1990s. The current PRRSV vaccines are not very effective largely due to heterogeneic nature of the virus. The major envelope protein, GP5, exposes outside the virion, induces neutralizing antibodies, and thus is a primary target for developing a subunit vaccine. In this study, we report a process for purification of GP5 protein from native virions of PRRSV propagated in MARC-145 cells. PRRSV virions were first purified and concentrated through sucrose cushion ultracentrifugation. GP5 protein was subsequently solubilized with Triton X-100 detergent for further processing. Cation exchange chromatography (CEX) was utilized for partial fractionation of GP5, although the viral nucleocapsid protein (N) was a major impurity in CEX elution fractions. During a second chromatographic step, hydrophobic interaction chromatography (HIC) further purified GP5 protein by means of a two-stage elution scheme. Pure GP5 protein was eluted from the HIC resin in the second HIC elution stage by Triton X-100 displacement; however the protein is present as a homodimeric/tetrameric aggregate. This process may be useful in PRRSV subunit vaccine development. PMID- 17913251 TI - Assessment of the diagnostic potential of immuno-RCA in 96-well ELISA plates for foot-and-mouth disease virus. AB - The need for fast and very early detection of foot-and-mouth disease virus (FMDV) infection has yielded different types of diagnostic tools over the past decades: whereas very sensitive techniques such as virus isolation (VI) and more recently also real-time RT-PCR can provide evidence for the presence of low virus quantities, VI requires additional confirmation of the nature of the virus strain and both techniques (currently) lack the ability for direct serotyping. The latter usually depends on ELISA, which is a far less sensitive method and may require virus culturing. This paper elaborates on experimental efforts towards the development of an 'immuno-rolling circle amplification (RCA)' assay in 96 well plates, the aim being to increase the sensitivity of immunological FMDV detection and serotyping by means of RCA. The study attempts to explain the encountered hurdles and the complexity of the different setups tested. Conclusively, immuno-RCA in 96-well plates as a reliable diagnostic assay for FMDV seems very difficult to achieve. PMID- 17913252 TI - Hydroxyurea enhances the activity of acyclovir and cidofovir against herpes simplex virus type 1 resistant strains harboring mutations in the thymidine kinase and/or the DNA polymerase genes. AB - Drug-resistant herpes simplex virus type 1 (HSV-1) recombinant strains harboring mutations in the thymidine kinase and/or the DNA polymerase genes were evaluated for their susceptibility to various antivirals in the presence of 25 microg/ml of hydroxyurea (HyU). The latter compound decreased the 50% inhibitory concentrations of acyclovir by 1.5-3.8-fold and that of cidofovir by 2.7-14.4 fold. However, HyU did not affect the susceptibilities of the various recombinant mutants to foscarnet. Hydroxyurea, a ribonucleotide reductase inhibitor, can increase the activity of nucleoside/nucleotide analogues against drug-resistant viruses. PMID- 17913253 TI - Mentally represented motor actions in normal aging II. The influence of the gravito-inertial context on the duration of overt and covert arm movements. AB - Here, we address the question of whether normal aging influences action representation by comparing the ability of 14 young (age: 23.6 +/- 2.1 years) and 14 older (age: 70.1 +/- 4.5 years) adults to mentally simulate arm movements under a varying dynamic context. We conducted two experiments in which we experimentally manipulated the gravity and inertial components of arm dynamics: (i) unloaded and loaded vertical arm movements, rotation around the shoulder joint, (ii) unloaded and loaded horizontal arm movements, rotations around the shoulder and elbow joints, in two directions (inertial anisotropy phenomenon). The main findings indicated that imagery ability was equivalent between the two groups of age for the unloaded arm movements, but better for the young than the older group, for the loaded arm movements. For the horizontal movements, we found better imagery ability for the young than the older adults for both movement directions and loads. Finally, young and old adults showed low (<8%)-temporal variability for both overt and covert arm movements in all conditions. Our findings showed a specific decline of action representation in the aging brain and suggest that internal models of action become imprecise with advance in age. This is not exact to say that there is a severe impairment of motor prediction in old adults as they can mentally represent their arm movements with high-temporal consistency. Finally, we propose that motor imagery could be used as a therapeutic tool for motor rehabilitation in aged adults. PMID- 17913254 TI - Estradiol and corticosterone independently impair spatial navigation in the Morris water maze in adult female rats. AB - The independent effects of ovarian and adrenal hormones on spatial place learning were examined in male and female Long-Evans hooded rats. Experimental groups received bilateral ovariectomy (females only) and adrenalectomy (both sexes), followed by hormone administration according to a predetermined schedule. Spatial and reversal training in the Morris water maze were used to measure behavioural performance in locating a hidden platform. General proficiency and strategies use were assessed using search times and time spent in the periphery, respectively. The number of direct and circle swims to the platform was used to assess memory for the location of the hidden platform. Experiment 1 investigated the roles of estradiol and progesterone in spatial navigation in the absence of high levels of adrenal steroids. The female group that received estradiol alone showed longer search times, greater periphery swimming, and fewer direct and circle swims to the target than all other female groups. Experiment 2 investigated the role of corticosterone (CORT) in spatial navigation in the absence of ovarian hormones. Male and female rats that received acute matched doses of exogenous CORT were equally impaired during spatial training. During reversal training, the impairment in search time, periphery swimming, and direct and circle swims persisted in the female CORT group only. Analysis of serum CORT levels in the male and female experimental groups revealed no significant differences. These data suggest that estradiol and CORT can independently impair acquisition of spatial navigation skills and strategies use in adult female rats. PMID- 17913256 TI - Neural mechanisms of voluntary and involuntary recall: a PET study. AB - Neuropsychological and neuroimaging studies on episodic memory retrieval have primarily focused on volitional memory tasks. However, some conscious memories arise involuntarily, i.e. without a strategic retrieval attempt, yet little is known about the neural network underlying involuntary episodic memory. The aim of this study was to determine whether voluntary and involuntary recall are mediated by separate cortical networks. We used positron emission tomography (PET) to measure changes in regional cerebral blood flow (rCBF) in 12 healthy subjects during voluntary and involuntary cued recall of pictures and a control condition with no episodic memory requirements. Involuntary recall was elicited by using an incidental memory task. Compared to the control condition, voluntary and involuntary recall were both associated with significant regional cerebral blood flow (rCBF) increases in posterior cingulate gyrus (PCG; BA 23), left precuneus (BA 7), and right parahippocampal gyrus (BA 35/36). In addition, rCBF in right dorsolateral prefrontal cortex (PFC; BA 8/9) and left precuneus (BA 7) was significantly larger during voluntary compared to involuntary recall, while rCBF was enhanced in left dorsolateral PFC (BA 9) during involuntary recall. The findings corroborate an association of the right PFC with a strategic component of episodic memory retrieval. Moreover, they show for the first time that it is possible to activate the medial temporal lobe, the PCG, and the precuneus, regions normally associated with retrieval success, without this strategic element. The relatively higher activity in precuneus during voluntary compared to involuntary recall suggests that activity in this region co-varies not only with retrieval success but also with retrieval intentionality. PMID- 17913255 TI - Pup exposure elicits hippocampal cell proliferation in the prairie vole. AB - The onset of parental behavior has profound and enduring effects on behavior and neurobiology across a variety of species. In some cases, mere exposure to a foster neonate (and a subsequent parental response) can have similar effects. In the present experiment, we exposed adult male and female prairie voles (Microtus ochrogaster) to two foster pups for 20 min and quantified cell proliferation in the dentate gyrus of the hippocampus (DG), medial amygdala (MeA) and cortical amygdala (CorA). Prairie voles are highly social rodents that typically display biparental care and spontaneous parental care when exposed to foster pups. Comparisons were made between the animals that responded parentally or non parentally towards the pups, as well as control conditions. Cell proliferation was assessed using injections of 5-bromo-2'-deoxyuridine (BrdU) and immunocytochemical localization of this marker. The phenotype of the cells was determined using double label immunofluoresence for BrdU and TuJ1 (a neuronal marker). An increase in cell proliferation in the DG was seen in animals exposed to pups. However, animals that responded non-parentally had a greater number of BrdU labeled cells in the DG compared to those that responded parentally. The majority of BrdU labeled cells co-expressed TuJ1 across all groups. These results demonstrate that exposure to a foster pup and the behavioral reaction to it (parental or non-parental) are associated with site-specific changes in cell proliferation. PMID- 17913257 TI - The Schistosoma mansoni protein Sm16/SmSLP/SmSPO-1 is a membrane-binding protein that lacks the proposed microtubule-regulatory activity. AB - Sm16/SmSLP/SPO-1 (Sm16) has been identified as a developmentally regulated protein that is released from specific glands of the Schistosoma mansoni parasite during skin penetration. Sm16 has been ascribed both anti-inflammatory activities and a functional similarity with the conserved cytosolic tubulin-binding protein stathmin/Op18. Here we used a cell line to confirm signal peptide-dependent secretion and to define the secreted form of Sm16 for production in E. coli. We present evidence from both in vitro experiments and studies on transfected human cells that refute any functional similarity with stathmin/Op18. Instead of an Op18-like activity, we found that targeting of Sm16 to the cytosol of human cells, which was achieved by ectopic expression of Sm16 lacking the signal peptide, results in a caspase-dependent apoptotic response. Interestingly, by analysis of recombinant preparations we found that the secreted form of Sm16 is a lipid bilayer-binding protein that efficiently binds to the surface of diverse cell types by a polyanion-independent mechanism, which results in uptake by endocytosis. While the significance of the pro-apoptotic activity exerted by cytosolic Sm16 remains unclear, the present findings on cell-surface-binding properties of Sm16 seems likely to be of functional relevance during skin penetration of the parasite. PMID- 17913258 TI - Ghrelin accelerates gastric emptying via early manifestation of antro-pyloric coordination in conscious rats. AB - Ghrelin is known to enhance gastric motility and accelerate gastric emptying of liquid and solid food in rats. As solid gastric emptying is regulated by the coordinated motor pattern between the antrum and pylorus (antro-pyloric coordination), we studied the correlation between solid gastric emptying and antro-pyloric coordination in response to ghrelin. Rats were given 1.5 g of solid food after a 24-h fasting. Immediately after the ingestion, ghrelin (0.4-8.0 microg/kg) or saline was administered by intraperitoneal (i.p.) injection. Ninety minutes after the feeding, rats were euthanized and gastric content was removed to calculate gastric emptying. To evaluate the antro-pyloric coordination, strain gauge transducers were sutured on the antrum and pylorus. The incidence of postprandial antro-pyloric coordination was compared between ghrelin-and saline injected rats. In saline-injected rats, gastric emptying was 58.3+/-3.7% (n=6). Ghrelin (4.0-8.0 microg/kg), accelerated gastric emptying. Maximum effect was obtained by ghrelin (4.0 microg/kg), which significantly accelerated gastric emptying to 77.4+/-3.7% (n=6, p<0.05). The number of antro-pyloric coordination 20-40 min after feeding was significantly increased in ghrelin-injected rats, compared to that of saline-injected rats (n=4, p<0.05). It is suggested that enhanced antro-pyloric coordination play an important role in accelerated solid gastric emptying induced by ghrelin. PMID- 17913259 TI - Effects of ghrelin on glucose-sensing and gastric distension sensitive neurons in rat dorsal vagal complex. AB - Ghrelin has been identified as the endogenous ligand of the growth hormone secretagogue receptor (GHS-R). Recent studies have shown that site-specific injection of ghrelin directly into the dorsal vagal complex (DVC) of rats is equally as sensitive in its orexigenic response to ghrelin as the arcuate nucleus of the hypothalamus (ARC). It is as yet unclear how circulating ghrelin would gain access to and influence the activity of the neurons in the DVC in which GHS receptors are expressed. In the present study, neuronal activity was recorded extracellularly in the DVC of anesthetized rats in order to examine the effects of ghrelin on the glucosensing neurons and the gastric distension (GD) sensitive neurons. The 82 neurons were tested with glucose, of which 26 were depressed by glucose and identified as glucose-inhibited (glucose-INH) neurons; 11 were activated and identified as glucose-excited (glucose-EXC) neurons. Of 26 glucose inhibited neurons examined for response to ghrelin, 23 were depressed, 1 was activated, and 2 failed to respond to ghrelin. Nine of 11 glucose-excited neurons were suppressed by ghrelin application, and the responses are abolished by the pretreatment with the GHS-R antagonist, [D-Lys-3]-GHRP-6. In addition, of 47 DVC neurons examined for responses to gastric distension (GD), 25 were excited (GD EXC), 18 were inhibited (GD-INH). 18 out of the 25 GD-EXC neurons were excited, whereas 15 out of 18 GD-INH neurons were suppressed by ghrelin. In conclusion, the activity of the glucosensing neurons in the DVC can be modulated by ghrelin, the primary effect of ghrelin on the glucose-INH and glucose-EXC neurons was inhibitory. Two distinct population of GD-sensitive neurons exist in the rat DVC: GD-EXC neurons are activated by ghrelin; the GD-INH neurons are suppressed by ghrelin. There is a diversity of effects of ghrelin on neuronal activity within the DVC, it is as yet unclear how this diversity in ghrelin's effects on cellular excitability contributes to ghrelin biological actions to influence food intake and gastric motility. PMID- 17913260 TI - Ghrelin deficiency does not influence feeding performance. AB - Ghrelin is an endogenous ligand for the growth hormone secretagogue receptor that is synthesized predominantly in the stomach. Previous studies demonstrated that ghrelin stimulates growth hormone release and food intake. These data suggested that antagonism of ghrelin could serve as a useful treatment for eating disorders and obesity. To study the role of endogenous ghrelin in feeding performance further, we generated ghrelin-deficient (ghrl(-/-)) mice. Unexpectedly, ghrl(-/-) mice exhibited normal growth, cumulative food intake, reproduction, histological characters, and serum parameters. There were no differences in feeding patterns between ghrl(+/+) and ghrl(-/-) mice. Ghrl(-/-) mice displayed normal responses to scheduled feedings as seen for ghrl(+/+) mice. Memory-related feeding performances of ghrl(-/-) mice were indistinguishable from ghrl(+/+) littermates. These data indicate that ghrelin is not critical for feeding performance. PMID- 17913261 TI - Expression of the human CMP-NeuAc:GM3 alpha2,8-sialyltransferase (GD3 synthase) gene through the NF-kappaB activation in human melanoma SK-MEL-2 cells. AB - To elucidate the mechanism underlying the regulation of human GD3 synthase gene expression in human melanoma SK-MEL-2 cells, we identified the promoter region of the human GD3 synthase gene. The 5'-rapid amplification of cDNA end (5'-RACE) using mRNA prepared from SK-MEL-2 cells revealed the presence of multiple transcription start sites of human GD3 synthase gene. Promoter analyses of the 5' flanking region of the human GD3 synthase gene using luciferase gene reporter system showed the strong promoter activity in SK-MEL-2 cells. Deletion study revealed that the region as the core promoter from -1146 to -646 (A of the translational start ATG as position +1) was indispensable for endogenous expression of human GD3 synthase gene. This region lacks apparent TATA and CAAT boxes but contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1 and NF-kappaB. Electrophoretic mobility shift assays using specific competitors, chromatin immunoprecipitation assay and site-directed mutagenesis demonstrated that only NF-kappaB element in this region is required for the promoter activity in SK-MEL-2 cells. These results indicate that NF-kappaB plays an essential role in the transcriptional activity of human GD3 synthase gene essential for GD3 synthesis in SK-MEL-2 cells. PMID- 17913262 TI - A review on the actual trends of insulin treatment in elderly with diabetes. AB - The diabetes of the elderly subjects has two forms: diabetes of long duration, manifesting itself in younger or medium ages, and senile diabetes, appearing above the age of 65 years. The diabetes of the elderly has usually only modest symptoms: it is not ketosic, but in spite of this, in order to avoid the chronic degenerative complications, it is important to maintain a good, even if not an optimal compensation. The therapeutic intervention cannot neglect a correct alimentary regime and a programmed physical activity in correlation with the clinical conditions of the patient. If the compensation is not achieved only with these tools, one can add oral antidiabetic treatments. In the elderly patients we usually observe primary or secondary failure of the oral antidiabetic treatments, and in such context we have to apply insulin treatment, even in cases of moderate glycometabolic decompensations. While we are waiting for the gene-therapy or the inhalatory insulin preparations, actually there are at disposal only the insulin analogs in rapid, slow and mixed forms. We propose two treatment schemes: (i) The first one consists of three administrations of rapid insulin with the meals, and on dose of slow insulin 2h after the last meal in the evening. (ii) The second scheme consists of one administration of rapid insulin at lunch, one administration of mixed insulin at dinner, with the addition of oral antidiabetics of peripheric action, in the morning and the evening. A better compliance can be obtained, being a fundamental aspect in the elderly diabetics, and a reduction of the number and severity of the hypoglycemia, which are the most important aspects in the elderly diabetes. PMID- 17913263 TI - Tongue-coating as risk indicator for aspiration pneumonia in edentate elderly. AB - Silent aspiration of oral microorganisms is a major cause of aspiration pneumonia. To establish oral hygiene criteria for the prevention of aspiration pneumonia in edentulous elderly persons, we investigated the relationship between presence of tongue-coating and number of oral bacteria in saliva and episodes of pneumonia. A total of 71 edentulous Japanese people aged 65 years or older living in nursing homes were enrolled in the study. A tongue plaque index (TPI) was used to evaluate quantity of tongue-coating, with TPI0 signifying no tongue-coating and TPI1 signifying presence of tongue-coating. Edentate elderly with TPI1 demonstrated significantly higher salivary bacterial counts than those with TPI0 (p<0.05). The number of elderly patients developing aspiration pneumonia was larger (p<0.005) in patients with TPI-based poor scores (average TPI>0.5) than in those with TPI-based good scores. The relative risk of developing pneumonia in the good tongue hygiene group compared with in the poor tongue hygiene group was 0.12, 95% confidence interval (CI): 0.02-0.9. The results demonstrate that tongue coating is associated with number of viable salivary bacterial cells and development of aspiration pneumonia, suggesting that tongue-coating is a risk indicator of aspiration pneumonia in edentate subjects. PMID- 17913264 TI - Predictors of elevated NT-pro-BNP in cardiovascular patients without acute heart failure. AB - AIM: B-type natriuretic peptides are recommended for evaluation of acute heart failure. Aim of this study was to identify predictors of pathologically elevated NT-pro-BNP in patients without acute heart failure. METHOD: NT-pro-BNP was measured in 486 inpatients with cardiovascular disease and the association with clinical and laboratory parameters was examined. Elevated NT-pro-BNP was defined according to rule-in cut-off values for acute heart failure: 450 pg/ml for age <50 years, 900 pg/ml for 50-75 years and 1800 pg/ml for >75 years. RESULTS: Multivariate analysis of variables assessable without echocardiography revealed fibrinogen, albumin, atrial fibrillation, history of heart failure and diuretic use as independent predictors of pathological NT-pro-BNP. The odds for pathological NT-pro-BNP were 2.5, 8.8, 27 and 41 in the presence of 1, 2, 3 and 4/5 predictors, respectively. After adjusting for cardiac dysfunction, fibrinogen, albumin, history of heart failure and atrial fibrillation still remained significant. CONCLUSION: Assessment of these parameters could help avoid wrong positive interpretation of NT-pro-BNP results. PMID- 17913265 TI - Catastrophic early drug eluting stents thrombosis and aspirin hypersensitivity. AB - Stent thrombosis is a feared complication of percutaneous coronary intervention. Promises and problems, late complications and early stent thrombosis have been reported after drug eluting stents implantation too. Moreover some patients with imperative cardiologic indications for combination therapy with aspirin and clopidogrel (stent placement and/or acute coronary syndrome) have a history of allergy to aspirin. We present a case of catastrophic early drug eluting stents thrombosis in a 79-year-old Italian woman with aspirin hypersensitivity. PMID- 17913266 TI - Active myocarditis in a patient with chronic active Epstein-Barr virus infection. AB - Chronic active Epstein-Barr virus (CAEBV) infection is characterized by chronic or recurrent infectious mononucleosis-like symptoms and the prognosis of CAEBV infection is quite poor. The incidence of myocarditis as a complication of EBV infection is not so high and it is unusual that heart failure appears as the initial symptom. However, it is very important to detect and treat chronic active myocarditis in the early phase of CAEBV infection because chronic active myocarditis disorganizes and decreases cardiomyocytes, resulting in the progression to heart failure. We report a case of a 45-year-old man with CAEBV infection for 5 years. Echocardiography revealed moderate left ventricular systolic dysfunction with mild pericardial effusion. Endomyocardial biopsies demonstrated massive lymphocytic infiltration with adjacent myocytolysis and necrosis of cardiomyocytes suggesting active myocarditis. Immunohistological analysis of biopsies revealed that the infiltrating cells were mainly T lymphocytes. And some of the infiltrating cells showed a positive signal for the EBV-encoded small nuclear RNA by in situ hybridization. Positron emission tomography using (18)F-fluoro-2-deoxyglucose ((18)F-FDG) performed revealed increased uptake of (18)F-FDG of whole left ventricular wall with mild heterogeneity. PMID- 17913267 TI - FFR guided PCI in a patient with dextrocardia and multivessel coronary artery disease. AB - Dextrocardia is a relatively rare congenital malposition in which the association with coronary artery disease is at the same frequency as in the general population. Previous reports have shown successful attempts at treating percutaneously patients with dextrocardia and coronary artery disease. We report the case of a patient with dextrocardia and multivessel coronary artery disease. In this case we highlight the use of physiologic measurements with a pressure wire, in order to select the significant coronary lesions to treat. PMID- 17913268 TI - Simultaneous subacute stent thrombosis of two sirolimus-eluting stents in a patient treated by ReoPro, thrombus aspiration and triple anti-platelet agents. AB - Effective anti-platelet treatment has a key role in the prevention of stent thrombosis in the drug-eluting stent (DES) era. However, despite of the use of anti-platelet agents, stent thrombosis occurs in approximately 1% of patients, with an increased likelihood of occurrence in high-risk patients or complex lesion subset of patients [1-4]. According to the previous report, triple anti platelet therapy (aspirin+clopidogrel+cilostazol) seemed to be more effective in preventing thrombotic complications after stenting than dual anti-platelet agent [5]. But, in this report, we present a patient with subacute stent thrombosis (ST) involving two different arteries simultaneously under the use of triple anti platelet regimen. PMID- 17913269 TI - Acute myocardial infarction in a patient with anomalous origin of the right coronary artery: depiction at whole-heart coronary magnetic resonance angiography and delayed-enhanced imaging. AB - A 71-year-old man was admitted to our hospital because of anterior chest pain. His electrocardiogram showed ST-segment depression and cardiac enzymes were normal. Non-ST-elevation acute myocardial infarction was suspected and whole heart magnetic resonance imaging was performed. Whole-heart coronary magnetic resonance angiography (MRA) showed an anomalous origin of the right coronary artery from the left sinus of Valsalva and delayed-enhanced imaging showed transmural hyperenhancement of the inferior wall. Coronary angiography revealed the anomalous origin of the right coronary artery (RCA) from the left sinus of Valsalva and occlusion in the proximal portion of the RCA. Coronary revascularization was achieved by intracoronary thrombolysis followed by stent implantation. Whole-heart coronary MRA and delayed-enhanced imaging allows simultaneous assessment of coronary artery anomaly and extent of myocardial infarction. PMID- 17913271 TI - Spontaneous coronary artery dissection in a professional body builder. AB - A 23-year-old male, a professional body builder, developed squeezing retrosternal chest pain following weight lifting. On examination, heart rate was 42 bpm and he had blood pressure of 70/50 mm Hg. An electrocardiogram was taken and showed inferior and right ventricular ST elevation myocardial infarction. Therefore, emergency coronary angiography was done and showed normal left coronaries. Right coronary artery angiogram revealed a dissection in the mid-part of the right coronary artery which was extended to the right ventricular branch. PMID- 17913272 TI - Spoilage of value-added, high-oxygen modified-atmosphere packaged raw beef steaks by Leuconostoc gasicomitatum and Leuconostoc gelidum. AB - Moisture-enhancing and marinating of meats are commonly used by the meat industry to add value to raw, retail products. Recently in Finland, certain value-added beef steak products have proven to be unusually susceptible to microbial spoilage leading to untoward quality deteriorations during producer-defined shelf-life. This study was conducted to evaluate the role of lactic acid bacteria (LAB) in the premature spoilage of value-added beef packaged under high-oxygen modified atmospheres. Spoilage was characterised by green discolouration and a buttery off odour. The predominant LAB in eight packages of spoiled, marinated or moisture enhanced beef steaks were identified by reference to a 16 and 23S rRNA gene restriction fragment length polymorphism pattern (ribotype) database. Leuconostoc gasicomitatum, Leuconostoc gelidum, Lactobacillus algidus, Lactobacillus sakei and Carnobacterium divergens were found to predominate in the LAB populations at numbers above 10(8) CFU/g. Inoculation of moisture-enhanced steaks with LAB strains and strain mixtures originating from the spoiled products demonstrated the spoilage potential of L. gasicomitatum and L. gelidum isolates. These two species produced green surface discolouration and buttery off-odours similar to these found in the spoiled, commercial products. PMID- 17913273 TI - Mycotoxins in rice. AB - Mycotoxin contamination in rice is usually lower as in wheat or corn. However, there are some reports that rice has been contaminated with mycotoxins such as aflatoxin B1, B2, G1, G2 (AFS), citrinin, deoxynivalenol (DON), fumonisin B1, B2, B3 (FMS), fusarenon-X (Fus.-X), nivalenol (NIV), ochratoxin A (OTA), sterigmatocystin (STE), and zearalenone. Rice in Japan is preserved in warehouses where moisture content and temperature are regulated. Therefore, mycotoxin contamination from post harvest fungal growth occurs very seldom. Trichothecenes, aflatoxins, and STE in rice were recently analyzed in our laboratory. In 1998, a typhoon struck before rice harvesting in Japan, and the unpolished rice was found to be stained brown. Samples were collected and analyzed for the presence of trichothecenes. Mycotoxins DON, Fus.-X, and NIV were detected and confirmed with GC-MS. The quantity of trichothecenes was determined using GC-ECD. STE is a carcinogenic mycotoxin produced by Aspergillus versicolor and some other fungi. STE contamination of rice was studied in our laboratory since 1973. GC-MS, LC-MS, LC-MS/MS, and LC-UV methods for STE determination were examined, giving good results for the LC-UV method using a photo diode array detector. Different techniques for the extraction of STE from rice were also studied. Finally, brown rice was ground, and the ground rice was extracted with acetonitrile-water. An Autoprep MF-A 1000 column was used to clean up AFS and STE. The cleaned-up extract was analyzed with HPLC-UV. Forty-eight brown rice samples were analyzed, and none of them were contaminated with STE. These rice samples were also analyzed for AFS and FMS, and none of the samples were contaminated. The Ministry of Agriculture, Forestry and Fisheries in Japan is making the appropriate Institutes develop analytical methods for mycotoxins and survey mycotoxin contamination on rice as well as wheat, corn, and some other cereals. PMID- 17913274 TI - Scaffold-free cartilage by rotational culture for tissue engineering. AB - Our objective was to investigate the hypothesis that tissue-engineered cartilage with promising biochemical, mechanical properties can be formed by loading mechanical stress under existing cell-cell interactions analogous to those that occur in condensation during embryonic development. By loading dedifferentiated chondrocytes with mechanical stress under existing cell-cell interactions, we could first form a scaffold-free cartilage tissue with arbitrary shapes and a large size with promising biological, mechanical properties. The cartilage tissue which constituted of chondrocytes and ECM produced by inoculated dedifferentiated chondrocytes to a high porous simple mold has arbitrary shapes, and did not need any biodegradable scaffold to control the shape. In contrast, scaffold-free cartilage tissue cultured under static conditions could not keep their shapes; it was fragile tissue. The possibility of scaffold-free organ design was suggested because the cartilage tissue increases steadily in size with culture time; indeed, the growth of cartilage tissue starting from an arbitrary shape might be predictable by mathematical expression. For tissue-engineered cartilage formation with arbitrary shapes, biochemical and mechanical properties, loading dedifferentiated chondrocytes with mechanical stress under existing cell-cell interactions has prominent effects. Therefore, our scaffold-free cartilage model loaded mechanical stress based on a simple mold system may be applicable for tissue-engineered cartilage. PMID- 17913275 TI - Topology of the global regulatory network of carbon limitation in Escherichia coli. AB - One fundamental shortcoming of biotechnological processes operating under carbon limiting conditions is the high-energy demand (maintenance) of the cells. Although the function of the central carbon metabolism in supplying precursors and energy for biosynthesis has been thoroughly characterized, its regulation and dynamic behaviour during carbon-limited growth has not yet been revealed. The current work demonstrates a time series of metabolic flux distributions during fed-batch cultivation of Escherichia coli K-12 W3110 applying a constant feed rate. The fluxes in glycolysis, pentose phosphate pathway and biosynthesis fell significantly, whereas TCA cycle fluxes remained constant. The flux redistribution resulted in an enhanced energy generation in the TCA cycle and consequently, in a 20% lower biomass yield. The intracellular alarmones ppGpp and cAMP accumulated in large quantities after the onset of nutrient limitation, subsequently declining to basal levels. The network topology of the regulation of the central metabolic pathways was identified so that the observed metabolic and regulatory behaviour can be described. This provides novel aspects of global regulation of the metabolism by the cra, crp and relA/spoT modulons. The work constitutes an important step towards dynamic mathematical modelling of regulation and metabolism, which is needed for the rational optimization of biotechnological processes. PMID- 17913276 TI - Lipoplex morphologies and their influences on transfection efficiency in gene delivery. AB - Cationic lipid-mediated gene transfer is widely used for their advantages over viral gene transfer because it is non-immunogenic, easy to produce and not oncogenic. The main drawback of the application of cationic lipids is their low transfection efficiency. Many reports about transfection efficiency of cationic lipids have been published in recent years. In this review, the current status and prospects for transfection efficiency of different morphologies of lipoplexes are discussed. High transfection activity will be acquired for H(C)(II) structure when membrane fusion is dominant, but when serum is present L(C)(alpha) lipoplexes show great superiority for their inhibition dissociation by serum during lipoplexes transporting. Increasing DOPE often gains high activity for the H(C)(II) structure promoted by DOPE. High lipofection will be gained from large lipoplexes when endocytosis is dominant, because large particles facilitate membrane contact and fusion. We suggest morphologies of lipoplex should be characterized at two levels, lipoplex size and self-assemble structures of lipoplexes, and understanding these would be very important for scientists to prepare novel cationic lipids and design novel formulations with high transfection efficiency. PMID- 17913277 TI - Reducing waist circumference by at least 3 cm is recommended for improving metabolic syndrome in obese Japanese men. AB - OBJECTIVE: We investigated the link between a reduction in waist circumference and metabolic syndrome. METHODS: 105 obese Japanese men were enrolled in this study with a 1-year follow-up. Anthropometric and body composition parameters, i.e. height, body weight, body fat percentage, waist circumference and hip circumference, blood pressure, triglyceride, HDL cholesterol and blood sugar, were evaluated. Metabolic syndrome was diagnosed using criteria developed in Japan. RESULTS: After a 1-year follow-up, the parameters of metabolic syndrome were significantly improved. The prevalence of metabolic syndrome was significantly reduced in subjects with at least 3 cm of waist circumference reduction (Group R). However, in subjects without such reduction (Group C), the prevalence of metabolic syndrome was similar to baseline levels. The prevalence of abdominal obesity, hypertension and dyslipidemia was also significantly reduced in Group R. In addition, there were remarkable differences of delta triglyceride (delta represents positive changes in parameters) and delta HDL cholesterol between Group R and Group C. CONCLUSION: At least 3 cm of waist circumference reduction may be beneficial for improving metabolic syndrome in obese Japanese men. PMID- 17913278 TI - Association of polymorphisms in the insulin-degrading enzyme gene with type 2 diabetes in the Korean population. AB - Insulin-degrading enzyme (IDE) is a metalloproteinase which degrades insulin and terminates its action. Homologous deletion of IDE gene resulted in hyperinsulinemia and glucose intolerance in a rat model of type 2 diabetes mellitus. Several genetic association studies examined IDE as a susceptibility gene for type 2 diabetes in European descents. Here we investigated the genetic association of IDE polymorphisms with the risk of type 2 diabetes and its related phenotypes in the Korean population. Among six single nucleotide polymorphisms analyzed, g.-179T>C (OR=1.73, P=0.04), and g.IVS18+99G>A (OR=1.23, P=0.02) revealed borderline association with increased risk of type 2 diabetes. Combining our results with previous data obtained from the European population, g.-179T>C (OR=1.11, P=0.03), and g.IVS24-64A>T (OR=1.18, P=0.005) showed significant association with type 2 diabetes. Haplotype consisting of common alleles of the six polymorphisms was associated with decreased risk of type 2 diabetes (OR=0.82, P=0.02). However, none of the polymorphisms was significantly associated with metabolic phenotypes. We can conclude that variations in IDE might contribute to diabetes susceptibility in the Korean population. PMID- 17913279 TI - The cost of in-patient care in Western Australia in the last years of life: a population-based data linkage study. AB - OBJECTIVES: This study aimed to explore patterns of health expenditure for in patient care in the last 3 years of life so as to understand how age and time to death contribute to health-care expenditure. METHOD: Records of all deaths occurring in Western Australia from 1997 to 2000 inclusive were extracted from the WA mortality register and linked to records from the hospital morbidity data system (HMDS) via the WA Data Linkage System. Inflation adjusted hospital costs were assigned to all in-patient events occurring within 3 years of death from five major causes of death using DRG costing information. RESULTS: Prior to the last 5 months of life the mean cost of hospitalisation was positively associated with age; however, the magnitude of the cost increase in the last 5 months of life was inversely related to age such that the cost in the last month of life was similar across age groups. CONCLUSION: The finding that increased costs are associated with proximity to death, but that the magnitude of the increase is inversely associated with age, has implications for the ongoing debate about whether proximity to death or age is the dominant driver of health-care costs. The results of this study suggest that models forecasting future health-care expenditure should take into account the interaction of age, time to death and cause of death. In addition, we propose that due to the differences observed across causes of death it may be that a single general population model may not be capable of fully capturing the relationship and that this may be why the debate regarding age and time to death has yet to be resolved in the literature. PMID- 17913280 TI - Polymorphisms in nucleotide excision repair genes, smoking and intake of fruit and vegetables in relation to lung cancer. AB - Polymorphisms in nucleotide excision repair genes have been associated with risk for lung cancer. We examined gene-environment interactions in relation to lung cancer in 430 cases and 790 comparison persons identified within a prospective cohort of 57,053 persons. We included polymorphisms in the XPC, XPA and XPD genes involved in the nucleotide excision DNA repair pathway and analysed possible interactions with smoking and dietary intake of fruit and vegetables in relation to risk for lung cancer. We found that intake of fruit was associated with lower risk for lung cancer only among carriers of the XPA A23G variant genotype. The incidence rate ratio for lung cancer was 0.60 (95% confidence interval: 0.43 0.84; p=0.003) per 50% increase in fruit intake. No convincing interactions were detected between the polymorphisms and smoking. PMID- 17913281 TI - Non-small cell lung carcinoma of the superior sulcus: favourable outcomes of combined modality treatment in carefully selected patients. AB - The combination of radiotherapy and concurrent chemotherapy followed by surgery (trimodality treatment) is currently regarded as optimal treatment for non-small cell lung cancer of the superior sulcus (SST) or Pancoast tumour. The possibility to administer intensive combined modality treatment is influenced by tumour stage, comorbidity and performance status of these patients, and therefore a strict patient selection is necessary. This study focuses on patient selection and its results. We retrospectively evaluated choices of treatment and outcome of all patients with SST treated in the Netherlands Cancer Institute from 1994 to 2004. After identification of patients with SST in registration databases, the following characteristics were analyzed: symptoms, comorbidity, tumour stage, treatment characteristics, toxicity, local control, disease-free and overall survival. Fifty-two patients, 37 men and 15 women, were identified. They were diagnosed with stage IIB (27%), stage IIIA (8%), stage IIIB (42%) and stage IV (23%). Twelve patients after induction (chemo)radiotherapy underwent surgical resection. In eight patients a pathologic complete response was found. The 2- and 5-year survival after induction treatment and surgery was 75 and 39%, respectively. Other patients did not receive surgical treatment because of stage IV disease (n=12), comorbidity (n=8), irresectability (extensive tumour growth and/or persisting N2-3 status; n=14) or insufficient response to induction treatment (n=6). Eleven patients were treated with concurrent chemoradiotherapy (5-year survival 20%) and 17 patients with (sequential) radiotherapy and/or chemotherapy (5-year survival 6%). Local recurrence rates were 0% after induction treatment and surgical resection, 32% after concurrent chemoradiotherapy and 72% after (sequential) radiotherapy and/or chemotherapy. In conclusion, only 30% of M0 patients with SST were eligible for combined modality treatment followed by surgery. In this subgroup, concurrent chemoradiotherapy followed by surgery was associated with excellent local control and acceptable survival. PMID- 17913282 TI - Usefulness of FDG-PET for early prediction of the response to gefitinib in non small cell lung cancer. AB - Increased tumor uptake of (18)F-fluorodeoxyglucose (FDG) measured by positron emission tomography (PET) reflects glucose metabolism and proliferative activity of tumor cells. We conducted a study to assess the usefulness of FDG-PET for early prediction of the response to gefitinib, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), in advanced non-small cell lung cancer (NSCLC). Five NSCLC patients underwent FDG-PET to evaluate changes in FDG uptake at day 2 and 4 weeks after the initiation of gefitinib therapy compared with FDG-PET prior to therapy. FDG uptake was evaluated as the maximum standardized uptake value (SUVmax) of the target lesions, which were assessable by conventional CT. Based on the CT evaluation, two patients exhibited a partial response (PR), two patients had stable disease (SD) with a minor response, and one patient had progressive disease (PD). In patients with PR and SD, SUVmax decreased by 61+/-18% (standard deviation) and 59+/-12%, respectively, on day 2, and by 26+/-6 and 43+/-10%, respectively, at 4 weeks after the initiation of gefitinib. Two patients with SD had decreased FDG uptake within 2 days of initiation of therapy, and achieved progression-free survival (PFS) of more than 12 months. In contrast, SUVmax increased up to 153+/-21% at 2 days and 232+/-73% at 4 weeks in a patient with PD. The present preliminary study suggests that FDG PET may be able to predict response to gefitinib in the early stage of therapy in patients with advanced NSCLC and may have a potential prognostic role. PMID- 17913283 TI - Performance evaluation of granular iron for removing hexavalent chromium under different geochemical conditions. AB - Long-term column experiments were conducted under different geochemical conditions to estimate the longevity of Fe 0 permeable reactive barriers (PRBs) treating hexavalent chromium (Cr(VI)). Secondary carbonate minerals were precipitated, and their effects on the performance, such as differences in the mechanism for Cr removal and the changes in system hydraulics, were assessed. Sequestration of Cr(VI) occurred primarily by precipitation of Fe(III)-Cr(III) (oxy)hydroxides. Trace amounts of Cr were observed in iron hydroxy carbonate presumably due to substitution of Cr3+ for Fe3+. The formation of Fe(III)-Cr(III) (oxy)hydroxide greatly decreased the reactivity of the Fe 0 and thus resulted in migration of the Cr removal front. Carbonate minerals did not appear to contribute to further passivation with regard to reactivity toward Cr removal; rather, the column receiving high contents of dissolved calcium carbonate showed slightly enhanced Cr removal by means of a higher corrosion rate of Fe 0 and because of sequestration by an iron hydroxy carbonate. Precipitation of carbonates, however, governed other geochemical parameters. The porosity and hydraulic conductivity in the column receiving high contents of dissolved calcium carbonate did not indicate a great loss in system permeability because the accumulation of carbonates declined as the Fe 0 was passivated over time. However, the accumulated carbonates and associated Fe(III)-Cr(III) (oxy)hydroxide could cause problems because the presence of these solids resulted in a decline in flow rate after about 1400 pore volumes of operation. PMID- 17913284 TI - A calcite permeable reactive barrier for the remediation of Fluoride from spent potliner (SPL) contaminated groundwater. AB - The use of calcite (CaCO3) as a substrate for a permeable reactive barrier (PRB) for removing fluoride from contaminated groundwater is proposed and is illustrated by application to groundwater contaminated by spent potliner leachate (SPL), a waste derived from the aluminium smelting process. The paper focuses on two issues in the implementation of calcite permeable reactive barriers for remediating fluoride contaminated water: the impact of the groundwater chemical matrix and CO2 addition on fluoride removal. Column tests comparing pure NaF solutions, synthetic SPL solutions, and actual SPL leachate indicate that the complex chemical matrix of the SPL leachate can impact fluoride removal significantly. For SPL contaminant mixtures, fluoride removal is initially less than expected from idealized, pure, solutions. However, with time, the effect of other contaminants on fluoride removal diminishes. Column tests also show that pH control is important for optimizing fluoride removal with the mass removed increasing with decreasing pH. Barrier pH can be regulated by CO2 addition with the point of injection being critical for optimising the remediation performance. Experimental and model results show that approximately 99% of 2300 mg/L fluoride can be removed when CO2 is injected directly into the barrier. This can be compared to approximately 30-50% removal when the influent solution is equilibrated with atmospheric CO2 before contact with calcite. PMID- 17913285 TI - Treatment with salicylic acid decreases the effect of cadmium on photosynthesis in maize plants. AB - The present study investigated the possible mediatory role of salicylic acid (SA) in protecting photosynthesis from cadmium (Cd) toxicity. Seeds of maize (Zea mays L., hybrid Norma) were sterilized and divided into two groups. Half of the seeds were presoaked in 500 microM SA solution for only 6h, after which both groups were allowed to germinate for 3d and were then grown for 14d in Hoagland solution at 22/18 degrees C in a 16/8-h light/dark period and 120 micromolm(-2)s(-1) PAR. All seedlings (without H(2)O and SA controls) were transferred to Cd-containing solutions (10, 15, and 25 microM) and grown for 14d. The rate of CO(2) fixation and the activity of ribulose 1,5-bisphosphate carboxylase (RuBPC, EC 4.1.1.39) and phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) were measured. Changes in the levels of several important parameters associated with oxidative stress, namely H(2)O(2) and proline production, lipid peroxidation, electrolyte leakage, and the activities of antioxidative enzymes (superoxide dismutase (SOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), catalase (CAT, EC 1.11.1.6), and guaiacol peroxidase (POD, EC 1.11.1.7)) were measured. Exposure of the plants to Cd caused a gradual decrease in the shoot and root dry weight accumulation, with the effect being most pronounced at 25 microM Cd. Seed pretreatment with SA alleviated the negative effect of Cd on plant growth parameters. The same tendency was observed for the chlorophyll level. The rate of CO(2) fixation was lower in Cd-treated plants, and the inhibition was partially overcome in SA pretreated plants. A drop in the activities of RuBPC and PEPC was observed for Cd treated plants. Pretreatment with SA alleviated the inhibitory effect of Cd on enzyme activity. Proline production and the rates of lipid peroxidation and electrolyte leakage increased in Cd-treated plants, whereas the values of these parameters were much lower in SA-pretreated plants. Treatment of plants with Cd decreased APX activity, but more than doubled SOD activity. Pretreatment with SA caused an increase in both APX and SOD activity, but caused a strong reduction in CAT activity. The data suggest that SA may protect cells against oxidative damage and photosynthesis against Cd toxicity. PMID- 17913286 TI - Involvement of cyclic GMP in phytochrome-controlled flowering of Pharbitis nil. AB - Light is one of the most important environmental factors influencing the induction of flowering in plants. Light is absorbed by specific photoreceptors- the phytochromes and cryptochromes system--which fulfil a sensory and a regulatory function in the process. The absorption of light by phytochromes initiates a cascade of related biochemical events in responsive cells, and subsequently changes plant growth and development. Induction of flowering is controlled by several paths. One is triggered by the guanosine-3':5'-cyclic monophosphate (cGMP) level. Thus, the aim of our study was to investigate the role of cGMP in phytochrome-controlled flowering. It is best to conduct such research on short-day plants because the photoperiodic reactions of only these plants are totally unequivocal. The most commonly used plant is the model short day plant Pharbitis nil. The seedlings of P. nil were cultivated under special photoperiodic conditions: 72-h-long darkness, 24-h-long white light with low intensity and 24-h-long inductive night. Such light conditions cause a degradation of the light-labile phytochrome. Far red (FR) treatment before night causes inactivation of the remaining light-stable phytochrome. During the 24-h long inductive darkness period, the total amount of cGMP in cotyledons underwent fluctuations, with maxima at the 4th, 8th and 14th hours. When plants were treated with FR before the long night, fluctuations were not observed. A red light pulse given after FR treatment could reverse the effect induced by FR, and the oscillation in the cGMP level was observed again. Because the intracellular level of cGMP is controlled by the opposite action of guanylyl cyclases (GCs) and phosphodiesterases (PDEs), we first tested whether accumulation of the nucleotide in P. nil tissue may be changed after treatment with a GC stimulator or PDE inhibitor. Accumulation of the nucleotide in P. nil cotyledons treated with a stimulator of cGMP synthesis (sodium nitroprusside) was markedly (approximately 80%) higher. It was highest in the presence of dipyridamole, whereas 3-isobutyl-1 methylxanthine did not significantly affect cGMP level. These results show that the analysed compounds were able to penetrate the cotyledons' tissue, and that they influenced enzyme activity and cGMP accumulation. FR light applied at the end of the 24-h-long white light period inhibited flowering. Exogenous cGMP added on cotyledons could reverse the effect of FR, especially when the compound was applied in the first half of the long night. Flowering was also promoted by exogenous application of guanylyl cyclase activator and phosphodiesterase inhibitors, and in particular dipyridamole. The results obtained suggest that an endogenous cGMP system could participate in the mechanism of a phytochrome controlled flowering in P. nil. PMID- 17913287 TI - Determination of phenolic content and antioxidant activity of extracts obtained from Rosmarinus officinalis' calli. AB - Rosmarinus officinalis is widely found in the lands of Aegean and Mediterranean regions of Turkey. Stem explants of very young shoots were cultured in both woody plant medium (WPM) and Murashige and Skoog (MS) media supplemented with 7g/L agar, 30g/L sucrose, and 1 and 3mg/L naphthaleneacetic acid (NAA) for callus initiation. Induced calli were subcultured 4 times with intervals of 7-10 days. MS medium supplemented with 1mg/L NAA proved to be the best medium for the production of callus (65.0%) among the samples tested. The lyophilized calli were subjected to solvent extraction. Active constituents of 8 calli extracts were analyzed by HPLC, and rosmarinic acid (RA) was determined to be the primary compound. Calli cultivated in WPM supplemented with 1mg/L NAA and extracted at 50 degrees C, yielded the highest amount of RA (34.4mg/g dry weight). Moreover, antioxidant activity of calli extracts was determined using a number of in vitro assays, including total phenol assay, DPPH radical scavenging activity (RSA), and trolox equivalent antioxidant capacity (TEAC). On the basis of the current findings, we conclude that WPM supplemented with 1mg/L NAA yields higher phenolic content as well as higher antioxidant activity. PMID- 17913288 TI - The protective role of selenium in rape seedlings subjected to cadmium stress. AB - The effect of selenium (Se) on rape (Brassica napus) seedlings subjected to cadmium (Cd) stress was studied in vitro by investigating plant growth and changes in fatty acid composition, activity of antioxidative enzymes and DNA methylation pattern. Physiological experiments were carried out on seedlings cultured for 2 weeks on Murashige-Scoog (MS) media with Cd concentrations of 0, 400 and 600 microM, and on corresponding media supplied with Se (2 microM). Exposure to increasing Cd concentrations reduced the fresh weight of the upper part (hypocotyls+cotyledons) of the seedlings more strongly than that of the root system, which was accompanied by higher Cd accumulation in these tissues. In the upper part, Cd exposure led to significant changes in the biochemical parameters: fatty acid unsaturation of plasmalemma decreased, the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPOX) diminished and that of ascorbate peroxidase (APX) increased. In contrast, the roots showed an increase in fatty acid unsaturation and in the activity of antioxidative enzymes. In both parts of rape seedlings H(2)O(2) level and lipid peroxidation increased. Se addition to medium considerably reversed the Cd-induced decrease in fresh mass as well as the changes in lipid unsaturation and peroxidation. Se applied separately or in combination with Cd did not significantly affect the activity of antioxidative enzymes in the roots, but diminished it in the upper part. Moreover, the presence of Se in medium prevented changes in the DNA methylation pattern triggered in rape seedlings by high Cd concentrations. Two possible mechanisms for the action of Se were considered: (1) removal of Cd from metabolically active cellular sites, and (2) reduction of oxygen radicals. PMID- 17913289 TI - CO2 fixation and chlorophyll a fluorescence in leaves of Ramonda serbica during a dehydration-rehydration cycle. AB - Changes in CO(2) photo-assimilation and PSII photochemical efficiency in Ramonda serbica leaves during a dehydration-rehydration cycle were examined. The rate of CO(2) photo-assimilation was greatly reduced during dehydration, but recovery was complete with rehydration when the relative water content of leaves reached values similar to those of well-hydrated, control leaves. The results showed that the response of R. serbica leaves to severe water stress involves two different mechanisms. In the first, CO(2) assimilation is limited by stomata closure that creates an excess proton concentration in the lumen and activates non photochemical quenching. This plays an important role in the mechanism of photoprotection by dissipation of excitation energy. When dehydration became severe and leaf RWC reached very low values, the electron transport rate (ETR) decreased markedly, while the capacity for regulatory mechanisms such as q(NP) (non-photochemical quenching) was greatly reduced. For severely dehydrated leaves of R. serbica, it appears that reactive oxygen species (ROS) formation is better prevented by mechanisms that quench chlorophyll triplet formation via lutein. PMID- 17913290 TI - Apoplastic antioxidative system responses to ozone stress in strawberry leaves. AB - Cell wall polysaccharides, pectin composition, as well as apoplastic superoxide dismutase and peroxidase activities were investigated in strawberry (Fragaria x ananassa) cultivars (cvs) Korona and Elsanta differing in their ozone sensitivity. Plants were exposed to 140-170 microg m(-3) ozone either short-term for 7 days or long-term for 2 months in order to investigate whether differences in ozone sensitivity were due to differences in the apoplastic antioxidative systems. Cell wall polysaccharides were increased after 7 days and 2 months of ozone stress. While water-soluble pectins, low methoxy pectinates and NaOH soluble pectinates were elevated after 7 days, their contents were unaffected (water-soluble pectins) or lower (low methoxy pectinates, NaOH-soluble pectinates) after 2 months. In cv. Elsanta, ozone treatment resulted in a significant reduction of superoxide dismutase activity after 7 days and 2 months, while it remained similar in cv. Korona. After 7 days, peroxidase activity was significantly higher in ozone-exposed leaves of cv. Korona, whereas after 2 months it was similar to or higher than in controls. Superoxide dismutase in cv. Korona detoxified ozone and its products in the apoplast, and the resulting elevated levels of H(2)O(2) were balanced within 7 days by an increase in peroxidase activity. Long-term peroxidase activity may not play a comparably significant role in ozone defence, but the increase in cell wall polysaccharides and cell wall thickness measured after 2 months, resulting in a decrease in specific leaf area, reflected structural modifications that limited activities of reactive oxygen species efficiently. In contrast, the reduction of superoxide dismutase activity in cv. Elsanta indicated a less efficient apoplastic radical scavenging system, at least during the first 7 days of ozone stress, which was accompanied by membrane leakage and contributed to accelerated leaf senescence. Long-term, the reduction of intercellular air space volume in leaves contributed to ozone tolerance of cv. Elsanta as in cv. Korona. PMID- 17913291 TI - Superoxide dismutase and total peroxidase activities in relation to drought recovery performance of mycorrhizal shrub seedlings grown in an amended semiarid soil. AB - We studied the effect of inoculation with a mixture of three arbuscular mycorrhizal (AM) fungi (Glomus intraradices Schenck & Smith, Glomus deserticola (Trappe, Bloss. & Menge) and Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe) and addition of a composted organic residue on plant growth, nutrient uptake, mycorrhizal colonisation and superoxide dismutase (SOD, EC 1.15.1.1) and total peroxidase (POX, EC 1.11.1.7) activities in shoots of Juniperus oxycedrus seedlings after well-watered, drought and recovery periods. The mycorrhizal inoculation and composted residue addition significantly increased the growth, foliar nutrients (N, P, K) and shoot water content of the plants, independent of the water regime. POX activity in control plants increased during drought (about 250% higher than under well-watered conditions) and returned to initial levels after re-watering. The seedlings inoculated with AM fungi showed the highest values of POX activity, followed by the plants grown in the amended soil, which varied little during the drought and recovery periods. Drought decreased the SOD activity in shoots of both J. oxycedrus seedlings inoculated with AM fungi and those grown with composted residue, but did not affect that of control plants. After re-watering, the SOD activity in mycorrhizal or residue-amended plants increased, showing values similar to control plants. PMID- 17913292 TI - Ethylene is required for elicitin-induced oxidative burst but not for cell death induction in tobacco cell suspension cultures. AB - The signal compound ethylene and its relationships with oxidative burst and cell death were analyzed in cultured tobacco cells treated with the proteinaceous elicitor quercinin. Quercinin belongs to the protein family of elicitins and was isolated from the soil-born oak pathogen Phytophthora quercina. It was shown to induce a dose-dependent oxidative burst in tobacco cell culture in concentrations from 0.05 to 0.5 nM, and subsequently, cell death. The characteristics of quercinin-induced cell death included both membrane damage and DNA fragmentation in tobacco cell culture. At higher quercinin concentrations (2 nM), H(2)O(2) formation and ethylene biosynthesis were inhibited. Ethylene at low concentrations proved to be necessary for induction and maintenance of H(2)O(2) production in tobacco cells treated with quercinin. It was demonstrated that external addition of inhibitors of ethylene biosynthesis such as alpha-amino-oxy acetic acid (AOA) and CoCl(2) also decreased or even inhibited the quercinin induced oxidative burst, but did not influence cell death induction. These results demonstrate evidence for a requirement of the plant hormone ethylene for the onset of the quercinin-induced oxidative burst. PMID- 17913293 TI - Fusarium oxysporum-induced oxidative stress and antioxidative defenses of yellow lupine embryo axes with different sugar levels. AB - This study was designed to investigate whether and to what extent oxidative stress is induced in embryo axes of Lupinus luteus L. cv. Polo inoculated with a necrotrophic fungus, Fusarium oxysporum and cultured on Heller medium for 96h. Four variants were compared: inoculated embryo axes cultured with 60mM sucrose (+Si) or without it (-Si), and non-inoculated embryo axes cultured with 60mM sucrose (+Sn) or without it (-Sn). After inoculation, an accumulation of stable free radicals and Mn2+ ions in +Si and -Si were detected by electron paramagnetic resonance. Concentrations of the radicals with g-values of 2.0052+/-0.0004 and 2.0029+/-0.0003 were generally higher in -Si than in +Si. Beginning at 24h after inoculation, in both +Si and -Si the concentrations of these ions decreased, but more strongly in -Si than in +Si. After inoculation, the activities of superoxide dismutase (SOD, EC 1.15.1.1) and catalase (CAT, EC 1.11.1.6) were higher in -Si than in +Si. SOD and CAT zymograms showed that the synthesis of new isoforms was induced after inoculation. Simultaneously, superoxide anions were assayed in embryo axes by using their specific indicator dihydroethidium (DHE). The DHE derived fluorescence was stronger and covered a much larger tissue area in +Si than in -Si. The respiration rate was generally much higher in +Si than in -Si. Electron micrographs revealed that, in contrast to -Si cells, +Si cells had numerous mitochondria with less reduced numbers of cristae and long sections of rough endoplasmic reticulum and Golgi bodies. These results indicate that different defensive strategies against F. oxysporum were induced depending on soluble sugar levels in yellow lupine embryo axes. PMID- 17913294 TI - Involvement of non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase in response to oxidative stress. AB - Glyceraldehyde-3-phosphate dehydrogenases catalyze key steps in energy and reducing power partitioning in cells of higher plants. Because non phosphorylating glyceraldehyde-3-phosphate dehydrogenase (NP-Ga3PDHase) is involved in the production of reductive power (NADPH) in the cytosol, its behavior under oxidative stress conditions was analyzed. The specific activity of the enzyme was found to increase up to 2-fold after oxidative conditions imposed by methylviologen in wheat and maize seedlings. Under moderate oxidant concentration, lack of mRNA induction was observed. The increase in specific activity would thus be a consequence of a significant stability of NP-Ga3PDHase. Our results suggest that the enzyme could be modified by oxidation of cysteine residues, but formation of disulfide bridges is dependent on levels of divalent cations and 14-3-3 proteins. The latter differential effect could be critical to relatively maintain energy and reductant levels in the cytoplasm of plant cells under oxidative stress. PMID- 17913295 TI - Overexpression of transcription factor OsLFL1 delays flowering time in Oryza sativa. AB - Flowering time is regulated by genetic programs and environment signals in plants. Genetic analysis of flowering time mutants is instrumental in dissecting the regulatory pathways of flower induction. Genotype W378 is a rice (Oryza sativa) late-flowering mutant selected from our collections of T-DNA insertion line. The T-DNA flanking gene in mutant W378 codes OsLFL1 (O. sativa LEC2 and FUSCA3 Like 1), a putative B3 DNA-binding domain-containing transcription factor. In wild-type rice OsLFL1 is expressed exclusively in spikes and young embryos, while in mutant W378 it is ectopically expressed. Introduction of OsLFL1-RNAi into mutant W378 successfully down-regulated OsLFL1 expression and restored flowering to almost normal time, indicating that overexpression of OsLFL1 confers late flowering for mutant W378. The flowering-promoting gene Ehd1 and its downstream genes are all down-regulated in W378. Thus, overexpression of OsLFL1 might delay the flowering of W378 by repressing the expression of Ehd1. PMID- 17913296 TI - Determinants of food rejection amongst school children. AB - This study investigates the role of food preferences in food rejection amongst primary school children. A total of 17 children (9- and 10-year-old) took part in group discussions from which elicited their preferences from 35 photographs of different foods and dishes. They also discussed their reasons for rejecting a specific meal that had been offered to them during a trip to France. The results showed that their food preferences played a limited role in their rejection. They reported to be disgusted by the meal, and this apparently stemmed from stereotype of French food associated with imagination and miscommunication. They also described aspects of the food as unfamiliar. PMID- 17913297 TI - The impact of personal resources and their goal relevance on satisfaction with food-related life among the elderly. AB - The study explored how actual resources, perceived levels of different types of resources and goal relevance of these resources affect older people's satisfaction with food-related life using a survey in eight European countries, where 3291 participants above 65 years of age and living in their own homes took part. Satisfaction with food-related life was measured using Satisfaction With Food-related Life (SWFL) scale developed by Grunert, Raats, Dean, Nielsen, Lumbers and The Food in Later Life Team. [(2007). A measure of satisfaction with food-related life. Appetite, 49, 486-493]. Results showed that older people rated the resources that they believed to have plentiful of as being highly relevant to achieve their goals. The individuals who rated the relevance and their level of different resources as high were also more satisfied with their food-related quality of life. Further, satisfaction with food-related life, as was expected, was predicted by income, health measures and living circumstances. However, the study also showed that perceived levels of other resources such as support of family and friends, food knowledge, storage facilities also added to the individuals' satisfaction with food-related life. In addition, the congruence between perceived level and relevance of a resource was also shown to add to people's satisfaction with food-related life, implying that older people's satisfaction with food-related life depends not only on the level of resources they think they have but also on their goals and how important they think these resources are to achieving their goals. PMID- 17913298 TI - The impact of emergency department crowding measures on time to antibiotics for patients with community-acquired pneumonia. AB - STUDY OBJECTIVE: We seek to determine the impact of emergency department (ED) crowding on delays in antibiotic administration for patients with community acquired pneumonia. METHODS: We performed a retrospective cohort study of adult patients admitted with community-acquired pneumonia from January 1, 2003, to April 31, 2005, at a single, urban academic ED. The main outcome was a delay (>4 hours from arrival) or nonreceipt of antibiotics in the ED. Eight ED crowding measures were assigned at triage. Multivariable regression and bootstrapping were used to test the adjusted impact of ED crowding measures of delayed (or no) antibiotics. Predicted probabilities were then calculated to assess the magnitude of the impact of ED crowding on the probability of delayed (or no) antibiotics. RESULTS: In 694 patients, 44% (95% confidence interval [CI] 40% to 48%) received antibiotics within 4 hours and 92% (95% CI 90% to 94%) received antibiotics in the ED. Increasing levels of ED crowding were associated with delayed (or no) antibiotics, including waiting room number (odds ratio [OR] 1.05 for each additional waiting room patient [95% CI 1.01 to 1.10]) and recent ED length of stay for admitted patients (OR 1.14 for each additional hour [95% CI 1.04 to 1.25]). When the waiting room and recent length of stay were both at the lowest quartiles (ie, not crowded), the predicted probability of delayed (or no) antibiotics within 4 hours was 31% (95% CI 21% to 42%); when both were at the highest quartiles, the predicted probability was 72% (95% CI 61% to 81%). CONCLUSION: ED crowding is associated with delayed and nonreceipt of antibiotics in the ED for patients admitted with community-acquired pneumonia. PMID- 17913299 TI - Emergency department crowding is associated with poor care for patients with severe pain. AB - STUDY OBJECTIVE: We study the impact of emergency department (ED) crowding on delays in treatment and nontreatment for patients with severe pain. METHODS: We performed a retrospective cohort study of all patients presenting with severe pain to an inner-city, teaching ED during 17 months. Poor care was defined by 3 outcomes: not receiving treatment with pain medication while in the ED, a delay (>1 hour) from triage to first pain medication, and a delay (>1 hour) from room placement to first pain medication. Three validated crowding measures were assigned to each patient at triage. Logistic regression was used to test the association between crowding and outcomes. RESULTS: In 13,758 patients with severe pain, the mean age was 39 years (SD 16 years), 73% were black, and 64% were female patients. Half (49%) of the patients received pain medication. Of those treated, 3,965 (59%) experienced delays in treatment from triage and 1,319 (20%) experienced delays from time of room placement. After controlling for factors associated with the ED treatment of pain (race, sex, severity, and older age), nontreatment was independently associated with waiting room number (odds ratio [OR] 1.03 for each additional waiting patient; 95% confidence interval [CI] 1.02 to 1.03) and occupancy rate (OR 1.01 for each 10% increase in occupancy; 95% CI 0.99 to 1.04). Increasing waiting room number and occupancy rate also independently predicted delays in pain medication from triage (OR 1.05 for each waiting patient, 95% CI 1.04 to 1.06; OR 1.18 for each 10% increase in occupancy; 95% CI 1.15 to 1.21) and delay in pain medication from room placement (OR 1.02 for each waiting patient, 95% CI 1.01 to 1.03; OR 1.06 for each 10% increase in occupancy, 95% CI 1.04 to 1.08). CONCLUSION: ED crowding is associated with poor quality of care in patients with severe pain, with respect to total lack of treatment and delay until treatment. PMID- 17913300 TI - Effect of emergency department crowding on time to antibiotics in patients admitted with community-acquired pneumonia. AB - STUDY OBJECTIVE: We hypothesize that emergency department (ED) volume and increased patient complexity are associated with lower quality of care, as measured by time to antibiotics for patients being admitted with community acquired pneumonia. METHODS: This was a cross-sectional study at a university tertiary care hospital ED. Community-acquired pneumonia patients admitted from the ED and discharged between January 2004 and June 2005 were reviewed by our institution for The Joint Commission's antibiotic timing core measure. Medical records were abstracted for patient age, sex, race, mode of transport, arrival time, triage acuity, inpatient level of care, and arrival-to-antibiotic administration times. Controlling for patient characteristics, multivariate logistic regression determined association of antibiotic administration within 4 hours of arrival, with total ED volume at the time of the community-acquired pneumonia patient's arrival, and with number of ED patients requiring admission at the time of arrival. RESULTS: Four hundred eighty-six patients were eligible for the study; antibiotic administration time was available for 405. Sixty-one percent of patients received antibiotics within 4 hours. Antibiotic administration within 4 hours was less likely with a greater number of patients (odds ratio 0.96 per additional patient; 95% confidence interval 0.93 to 0.99) and a greater number of patients ultimately admitted (odds ratio 0.93 per patient; 95% confidence interval 0.88 to 0.99) in the ED. The effect of additional patients was present below total ED capacity. CONCLUSION: As ED volume increases, ED patients with community-acquired pneumonia are less likely to receive timely antibiotic therapy. The effect of additional patients appears to occur even at volumes below the maximum bed capacity. Measures to ensure that quality targets are met in the ED should consider the impact of ED volume. PMID- 17913301 TI - Increased circulating urotensin II in cirrhosis: potential implications in liver disease. AB - Urotensin II (UII) is a potent vasoactive mediator which, through interaction with a specific G-protein coupled receptor, can result in either a vasoconstrictive or vasodilatory response. In addition to its effect upon vascular tone, UII possess mitogenic and fibrogenic potential. The influence of UII on vascular tone is to some degree both species-specific and disease specific. Increased circulating UII levels have been documented in subjects with liver cirrhosis although the significance of this finding with regards to the development of chronic liver disease and portal hypertension has yet to be fully elucidated. In this review we focus on the potential relevance of UII as a vasoactive mediator in the chronic liver disease population and postulate as to the site of overproduction of UII. PMID- 17913302 TI - 9-Methyl-beta-carboline up-regulates the appearance of differentiated dopaminergic neurones in primary mesencephalic culture. AB - beta-Carbolines (BCs) derive from tryptophan and its derivatives. They are formed endogenously in humans and mammals and occur inter alia in cooked meat and tobacco smoke. They have been detected in human brain, cerebrospinal fluid, and plasma. Up to now they were predominantly identified as compounds exhibiting neurotoxic actions. Since significantly higher amounts are present in parkinsonian patients, they are regarded as potential pathogenetic factors in Parkinson's disease. We identified for the first time a BC (9-methyl-BC; 9-me-BC) exerting neuroprotective and neuron-differentiating effects. Treatment of primary mesencephalic dopaminergic cultures with 9-me-BC inhibited the basal release of lactate dehydrogenase and reduced the number of cells stained with propidium iodide. Caspase-3 activity was decreased, the total protein content was unchanged and ATP content was increased. Furthermore, the expression of inflammation related genes was reduced. The number of differentiated dopaminergic neurones was significantly increased and a wide array of neurotrophic/transcription factors (Shh, Wnt1, Wnt5a, En1, En2, Nurr1, Pitx3) and marker genes (Th, Dat, Aldh1a1) decisive for dopaminergic differentiation was stimulated. Consistently, the dopamine content was slightly, although non-significantly, increased and the dopamine uptake capacity was elevated. An anti-proliferative effect was observed in human neuroblastoma SH-SY5Y cells which is consistent with a reduced incorporation of bromodesoxyuridine into the DNA of primary mesencephalic cells. Whether the additional dopaminergic neurones in primary culture derive from dopaminergic precursor cells, previously tyrosine hydroxylase negative dopaminergic neurones or are the result of a transdifferentiation process remains to be established. PMID- 17913303 TI - Genetic priming of a proinflammatory profile predicts low IQ in octogenarians. AB - The purpose of the study was to test the hypothesis that single nucleotide polymorphisms (SNPs) within interleukin (IL)-18, TNF-alpha, IL-6 and IL-10 gene promoter regions are risk factors for cognitive decline in healthy octogenarians, and to isolate the strongest inflammatory biomarkers of cognitive function in the peripheral blood. The Wechsler Adult Intelligence Scale was administered to 112 individuals at ages 80 and 85. An IL-18 haplotype was an independent risk factor of poor Performance IQ. The TNF-308GA genotype was related to individual declines in Verbal IQ, and the IL-10-592 CC genotype was related to better Verbal IQ at the age of 80. Circulating levels of TNF-alpha, sTNFRs, and IL-6 were negatively correlated with IQ at age 85 and less strongly to IQ at age 80 with activation of the TNF system as the strongest biomarker. In conclusion, SNPs related to high proinflammatory or low anti-inflammatory activity are independent risk factors of reduced cognitive function in octogenarians. Only the IL-18 haplotype was associated with inflammation in the peripheral blood and only with regard to circulating TNF-alpha. PMID- 17913305 TI - Oral vaccination with liposome-encapsulated recombinant fusion peptide of urease B epitope and cholera toxin B subunit affords prophylactic and therapeutic effects against H. pylori infection in BALB/c mice. AB - A new fusion peptide CtUBE of cholera toxin B subunit and Helicobacter pylori urease B subunit epitope was expressed in Escherichia coli. With this fusion peptide, an oral liposome vaccine against H. pylori infection was prepared and evaluated in BALB/c mice. Based on the results of urease tests, quantitation of culturable bacteria colonies in mice stomachs and histological identification of gastritis, the mice were protected significantly after intragastric vaccination with this CtUBE liposome vaccine, which increased the content levels of specific anti-urease serum IgG and mucosal IgA for both prophylactic and therapeutic vaccination protocols. These results showed that the fusion peptide CtUBE retained immunogenicity and could be used as antigen in the development of an oral vaccine against H. pylori infection. PMID- 17913306 TI - DNA vaccination with VP2 gene of very virulent infectious bursal disease virus (vvIBDV) delivered by transgenic E. coli DH5alpha given orally confers protective immune responses in chickens. AB - The efficacy of different doses of oral DNA vaccines carrying VP2 gene of vvIBDV delivered by E. coli DH5alpha was studied and compared with purified VP2 recombinant expression plasmid DNA vaccine injected intradermally and whole virus vaccine either from homologous virus or from commercial source. The recombinant plasmid pRc-VP2 was transformed in a non-pathogenic strain of E. coli, the DH5alpha and designated as EC/pRC-VP2. Oral immunization of maternal antibody free broiler chickens at 7 and 14-day-old with different dosages of EC/pRc-VP2 elicited specific humoral immune response as measured by ELISA. Protection in different groups was calculated through clinical signs, gross and histopathological lesions, bursa of Fabricius to body weight ratio, humoral and cellular immune responses and mortality in the chickens. Vaccination with EC/pRc VP2 at the dose rate of 10(9)CFU per chicken conferred 95.4% protection of the chickens against the challenge with homologous virulent field strain of vvIBDV. Protection afforded by attenuated vero cell adapted UAF-06 strain of vvIBDV was comparable (94%) to that by EC/pRc-VP2 and pRc-VP2 vaccines, which was significantly higher (P<0.05) than the protection provided by a commercial attenuated IBDV stain D-78 vaccine (D-78 vaccine was used as positive control due to its frequent use in the field for vaccination of poultry chickens) and other control groups in the study. The results revealed that DNA vaccines against IBDV may be successfully done by adopting bacterial-vectored oral delivery system and vaccination with homologous vvIBDV (UAF-06) conferred significantly higher protection as compared with imported non-homologous commercial IBDV vaccine. PMID- 17913307 TI - Evaluating the vaccine potential of an influenza A viral hemagglutinin and matrix double insertion DNA plasmid. AB - A DNA plasmid expressing both the influenza viral matrix protein (M1) and hemagglutinin (HA) (pHA/M1) as a potential vaccine candidate was investigated. Vaccination with pHA/M1 double insertion plasmids not only induced HA-specific protective antibodies, but also elicited HA and M1-specific CD8 T cell responses. Mice immunized with pHA/M1 dual expressing plasmid showed enhanced HA inhibition titer and increased CD69(+) CD8alpha(+) T cell response compared to groups that received either the vector or a mixture of both pHA and pM1 (pHA+pM1). Furthermore, pHA/M1 immunization resulted in improved protection against both homologous and heterologous challenges. PMID- 17913308 TI - A Yersinia pestis lpxM-mutant live vaccine induces enhanced immunity against bubonic plague in mice and guinea pigs. AB - The lpxM mutant of the live vaccine Yersinia pestis EV NIIEG strain synthesising a less toxic penta-acylated lipopolysaccharide was found to be avirulent in mice and guinea pigs, notably showing no measurable virulence in Balb/c mice which do retain some susceptibility to the parental strain itself. Twenty-one days after a single injection of the lpxM-mutant, 85-100% protection was achieved in outbred mice and guinea pigs, whereas a 43% protection rate was achieved in Balb/c mice given single low doses (10(3) to 2.5 x 10(4) CFU) of this vaccine. A subcutaneous challenge with 2000 median lethal doses (equal to 20,000 CFU) of fully virulent Y. pestis 231 strain, is a 6-10-fold higher dose than that which the EV NIIEG itself can protect against. PMID- 17913309 TI - Further evaluation of higher potency vaccines for early protection of cattle against FMDV direct contact challenge. AB - The effect of administering higher payload FMD vaccines 10 days prior to severe direct contact challenge on protection from clinical disease and sub-clinical infection was investigated in cattle using two antigen payloads (single strength and 10-fold). Regardless of antigen payload, vaccination was shown to significantly reduce the number of clinically infected animals, and significantly reduce virus excretion shortly after challenge, when compared with the unvaccinated group (P<0.05). Although FMDV transmission occurred from single strength vaccinated infected cattle to similarly vaccinated cattle held in indirect contact, no disease was induced in these animals. These studies further confirm that emergency vaccination does significantly reduce clinical disease and sub-clinical virus replication and excretion, particularly early post exposure, thereby reducing the possibility of transmission between animals and herds. To be most effective, however, the results also substantiate that time of vaccination prior to challenge significantly influences the number of animals becoming infected, so the decision to vaccinate should be made swiftly, to allow maximum opportunity for protective immunity to develop. PMID- 17913311 TI - Immunogenicity of a killed Leishmania vaccine with saponin adjuvant in dogs. AB - Cellular and humoral immune responses of dogs to a candidate vaccine, composed of Leishmania braziliensis promastigote protein plus saponin as adjuvant, have been investigated as a pre-requisite to understanding the mechanisms of immunogenicity against canine visceral leishmaniasis (CVL). The candidate vaccine elicited strong antigenicity related to the increases of anti-Leishmania IgG isotypes, together with higher levels of lymphocytes, particularly of circulating CD8(+) T lymphocytes and Leishmania chagasi antigen-specific CD8(+) T-lymphocytes. As indicated by the intense cell proliferation and increased nitric oxide production during in vitro stimulation by L. chagasi soluble antigens, the candidate vaccine elicited an immune activation status potentially compatible with effective control of the etiological agent of CVL. PMID- 17913310 TI - Safety and immunogenicity of a high dosage trivalent influenza vaccine among elderly subjects. AB - To improve immune responses to influenza vaccine, a trivalent inactivated vaccine containing 60 microg of the HA of each component (A/H3N2, A/H1N1, B) was compared to a licensed vaccine containing 15 microg of the HA of each. More local and systemic reactions were reported by subjects given the high dosage but only local pain and myalgias were significantly increased. The high dosage vaccine induced a higher frequency of serum antibody increases (> or =4-fold) in both hemagglutination-inhibiting (HAI) and neutralization tests for all three vaccine viruses in the total group as well as subjects vaccinated and those not vaccinated the previous year. Mean titers of antibody attained, the magnitude of antibody increases and the frequencies of persons with final HAI antibody titers > or =1:32, > or =1:64, and > or =1:128 were all greater for the high dosage group in both serologic tests, for all groups, and for all vaccine viruses. These increased immune responses should provide increased protection against influenza in the elderly. PMID- 17913312 TI - Access to vaccine technologies in developing countries: Brazil and India. AB - This study, conducted by visits, interviews, and literature search, analyzes how vaccine manufacturers in Brazil and India access technologies for innovative vaccines: through collaborations with academia and research institutions, technology transfer agreements with multinational corporations, public sector, or developing country organizations, or by importation and finishing of bulk products. Each has advantages and disadvantages in terms of speed, market, and ability to independently produce the product. Most manufacturers visited are very concerned about avoiding patent infringement, which might result in undeveloped or delayed products because of a lack of mastery of the patent landscape. Disregarding the patent picture could also threaten the market of a potential product. Although it is too soon to assess the effects of TRIPS on vaccine technology access in Brazil and India, a good understanding of intellectual property management will be useful. A case study on development of a new combination vaccine illustrates these findings. PMID- 17913313 TI - Anthrax vaccination in a military population before the war in Iraq: side effects and informed choice. AB - BACKGROUND: To assess any health consequences of the anthrax vaccination programme in UK Armed Forces deployed to Iraq. METHODS: Data were collected from two samples simultaneously. The first was 5302 randomly selected UK service personnel. The second was 607 service personnel involved in a longitudinal study, where pre-vaccination health had previously been collected. Both samples were offered the anthrax vaccination before they deployed to Iraq in 2003 and subsequently following their service in Iraq. Participants completed a detailed questionnaire, including a range of health outcomes, receipt of the anthrax vaccination and quality of choice. RESULTS: Seventy-two percent of the combined sample had the anthrax vaccination. Being a member of the Army, a commissioned officer or a reservist was associated with higher uptake. No differences in self reported health were observed between those who did and did not receive the vaccination. For participants who accepted the vaccination, we found an association between making an uninformed choice and adverse health. After adjustment for baseline health in the longitudinal cohort these associations remained significant. CONCLUSIONS: Anthrax vaccination used by the UK Armed Forces in preparation for the Iraq War has not resulted in adverse health outcomes. However, of those who did accept the vaccination, reported side effects were related to whether acceptance of vaccination was perceived to be informed. Improving the quality of choice may improve self-reported ill health. PMID- 17913314 TI - Cattle immune response to botulinum type D toxoid: results of a vaccination study. AB - Cattle botulism is a food-borne intoxication caused by the ingestion of preformed botulinum neurotoxins (BoNT) of serotypes B, C, or D. Protection in cattle against botulinum intoxication is based on the presence of specific serum neutralizing antibodies upon exposure. Outbreaks in vaccinated cattle have raised concerns about vaccine quality and efficacy. To this end, three different immunization protocols and the effect of maternal anti-BoNT/D antibodies, at the priming dose, were analyzed in 2-month-old dairy calves. Based on previously determined protective anti-BoNT/D antibody levels analyzed in field outbreaks, the immune response to type D toxoids was analyzed using an in-house ELISA system. Here we show that using the current vaccination strategy of using a priming dose in 2-month-old calves followed by booster doses after 4 weeks and annually thereafter, did not result in continuous protective levels of anti BoNT/D antibodies. As a result of this vaccination protocol, only 15-31% of cattle in parities 1-3 were protected at the time of the annual booster. Vaccination study in calves indicated that adding a 6-month booster dose to the current protocol resulted in continuous protective levels of anti-BoNT/D antibodies well above the cut-off protective levels. The presence of maternally derived anti-BoNT/D antibodies did not interfere with the immune response to toxoids that can be administered to 2-month-old calves. PMID- 17913315 TI - Determination of specific activity of (226)Ra, (232)Th and (40)K for assessment of radiation hazards from Turkish pumice samples. AB - The specific activity of (226)Ra, (232)Th and (40)K in 52 Turkish pumice samples collected from 11 geographical areas located in Central Anatolia, Eastern Anatolia, Mediterranean and Aegean regions was determined by gamma-ray spectrometry with a high-purity germanium (HPGe) detector. The specific activity of (226)Ra, (232)Th and (40)K ranged from 12.7+/-0.5 to 256.2+/-9.1Bqkg(-1) with a mean of 89.1+/-65.2Bqkg(-1), 12.3+/-1.0 to 237.9+/-12.2Bqkg(-1) with a mean of 87.0+/-61.4Bqkg(-1) and 300.1+/-5.5 to 1899.0+/-30.8Bqkg(-1) with a mean of 1211.9+/-419.8Bqkg(-1), respectively. Elemental concentrations were determined for U (from 1.0 to 20.7ppm with a mean of 7.2+/-5.3ppm), Th (from 3.0 to 58.6ppm with a mean of 21.4+/-15.1ppm) and K (from 1.0 to 6.1% with a mean of 3.9+/ 1.3%). The radium equivalent activity (Ra(eq)), the activity index, the emanation coefficient, the (222)Rn mass exhalation rate, the indoor absorbed dose rate and the effective dose rate were estimated for the radiation hazard of the natural radioactivity in all samples. The calculated mean Ra(eq) value was 306.6+/ 177.7Bqkg(-1) (54.6+/-5.5 to 737.6+/-49.0Bqkg(-1)) for all pumice samples. This value is lower than the recommended limit value of 370Bqkg(-1) for building raws and products. The emanation coefficient and the (222)Rn mass exhalation rate of all samples ranged from 29.4 to 42.9% with a mean of 36.2% and from 11.0 to 196.4microBqkg(-1)s(-1) with a mean of 73.5microBqkg(-1)s(-1), respectively. The mean indoor absorbed dose rate and the corresponding mean effective dose rate were 274.6+/-153.6nGyh(-1) (50.4-644.6nGyh(-1)) and 1.35+/-0.75mSvy(-1) (0.24 3.16mSvy(-1)), respectively. For all pumice samples the mean indoor absorbed dose rate is about three times higher than the population-weighted average of 84nGyh( 1), while the mean effective dose rate values except for PUM 05, PUM 06, PUM 10 and PUM 15 exceed the dose criterion of 1mSvy(-1). PMID- 17913316 TI - Radon releases from Australian uranium mining and milling projects: assessing the UNSCEAR approach. AB - The release of radon gas and progeny from the mining and milling of uranium bearing ores has long been recognised as a potential radiological health hazard. The standards for exposure to radon and progeny have decreased over time as the understanding of their health risk has improved. In recent years there has been debate on the long-term releases (10,000 years) of radon from uranium mining and milling sites, focusing on abandoned, operational and rehabilitated sites. The primary purpose has been estimates of the radiation exposure of both local and global populations. Although there has been an increasing number of radon release studies over recent years in the USA, Australia, Canada and elsewhere, a systematic evaluation of this work has yet to be published in the international literature. This paper presents a detailed compilation and analysis of Australian studies. In order to quantify radon sources, a review of data on uranium mining and milling wastes in Australia, as they influence radon releases, is presented. An extensive compilation of the available radon release data is then assembled for the various projects, including a comparison to predictions of radon behaviour where available. An analysis of cumulative radon releases is then developed and compared to the UNSCEAR approach. The implications for the various assessments of long-term releases of radon are discussed, including aspects such as the need for ongoing monitoring of rehabilitation at uranium mining and milling sites and life-cycle accounting. PMID- 17913317 TI - Propofol alone and combined with dexamethasone for the prevention of postoperative nausea and vomiting in adult Japanese patients having third molars extracted. AB - We did a prospective, randomised, double-blind study to evaluate the efficacy and safety of a small dose of propofol alone, and propofol combined with dexamethasone, for the prevention of postoperative nausea and vomiting in adult Japanese patients listed for third molars extractions. One hundred and twenty patients, 55 men and 65 women aged 17-48 years, were given placebo, propofol 0.5mg/kg, or propofol 0.5mg/kg plus dexamethasone 8 mg intravenously at the end of the operation. A standard general anaesthestic was used, including sevoflurane and nitrous oxide in oxygen. Patients' characteristics were comparable in all three groups. The numbers of patients who developed postoperative nausea and vomiting during the 24h after anaesthesia were 8 with propofol (p=0.04), 2 with propofol plus dexamethasone (p=0.001), and 16 with placebo. The antiemetic efficacy of propofol combined with dexamethasone was superior to that of propofol alone (p=0.04). There were no clinically important adverse events. We conclude that a small dose (0.5mg/kg) of propofol combined with dexamethasone 8 mg was more effective than propofol alone for the prevention of postoperative nausea and vomiting in adult Japanese patients having general anaesthesia for extractions of third molars. PMID- 17913318 TI - Toxaphene in minke whales (Balaenoptera acutorostrata) from the North Atlantic. AB - Toxaphene contamination of minke whales (Balaenoptera acutorostrata) from North Atlantic waters was examined for the first time. Total toxaphene and SigmaCHB (sum of 11 chlorobornanes) concentrations in blubber samples ranged from 170+/ 110 and 41+/-39 ng/g lipid weight (l.w.) for female minke whales from southeastern Greenland to 5800+/-4100 and 1100+/-780 ng/g l.w. for males from the North Sea, respectively. Very large variations in toxaphene concentrations among sampling areas were observed suggesting a spatial segregation of minke whales. However, much of the apparent geographical discrimination was explained by the seasonal fluctuation of animal fat mass. Patterns of CHBs in males revealed that recalcitrant CHBs were in higher proportions in animals from the more easterly areas than in animals from the more westerly areas. This trend may be influenced by the predominance of the US, over the European, input of toxaphene to North Atlantic waters. PMID- 17913319 TI - The predictive, convergent, and discriminant validity of perceived criticism: a review. AB - People's perceptions of criticism from another individual, known as perceived criticism (PC), have been found to predict symptom course, treatment outcome, and future relapse across a number of disorders, including depression, anxiety disorders, substance abuse, and schizophrenia. Some recent studies, however, have yielded conflicting results, making the overall pattern of findings difficult to interpret. This article reviews the findings on PC and presents a framework for understanding them, focused on predictive, convergent, and discriminant validity. The following questions are addressed: (1) What conclusions can be drawn about the prospective relationship of PC with symptom fluctuation and treatment response across disorders? (2) Does it matter whom respondents rate in terms of PC? (3) Is PC a valid construct for individuals from non-Western cultures? (4) Does the fact that the typical measure of PC consists of only 1 item limit its utility? (5) Is PC best viewed as an accurate reflection of another person's critical behavior, another index of overall satisfaction in an interpersonal relationship, some intraindividual variable (e.g., general sensitivity to criticism), or some combination of these factors? Based on the answers to these and other questions, recommendations for clinical practice and future research related to PC are made. PMID- 17913321 TI - Social capital, income inequality, and self-rated health in 45 countries. AB - There has been growing interest in the relationship between the social environment and health. Among the concepts that have emerged over the past decade to examine this relationship are socio-economic inequality and social capital. Using data from the World Values Survey and the World Bank, we tested the hypothesis that self-rated health is affected by social capital and income inequality cross-nationally. The merit of our approach was that we used multilevel methods in a larger and more diverse sample of countries than used previously. Our results indicated that, for a large number of diverse countries, commonly used measures of social capital and income inequality had strong compositional effects on self-rated health, but inconsistent contextual effects, depending on the countries included. Cross-level interactions suggested that contextual measures can moderate the effect of compositional measures on self rated health. Sensitivity tests indicated that effects varied in different subsets of countries. Future research should examine country-specific characteristics, such as differences in cultural values or norms, which may influence the relationships between social capital, income inequality, and health. PMID- 17913320 TI - Poverty, wealth inequality and health among older adults in rural Cambodia. AB - Little research exists on health determinants among adults living in economically deprived regions despite the fact that these areas comprise a good part of the world. This paper examines the distribution of wealth then tests associations between wealth inequality and a variety of health outcomes, among older adults, in one of the world's poorest regions--rural Cambodia. Data from the 2004 Survey of the Elderly in Cambodia are employed. Using a disablement framework to conceptualize health, associations between four health components and a wealth inequality measure are tested. The wealth inequality measure is based on an index that operationalizes wealth as ownership of household assets and household structural components. Results confirm difficult economic conditions in rural Cambodia. The lowest wealth quintile lives in households that own nothing, while the next quintiles are only slightly better off. Nevertheless, logistic regressions that adjust for other covariates indicate heterogeneity in health across quintiles that appear qualitatively similar, with the bottom quintiles reporting the most health problems. An exception is disability, which presents a U-shaped association. It is difficult to determine mechanisms behind the relationship using cross-sectional data, but the paper speculates on possible causal directions, both from wealth to health and vice-versa. The analysis suggests the ability to generalize the relationship between wealth inequality and health to extremely poor populations as a very small difference in wealth makes a relatively large difference with respect to health associations among those in meager surroundings. PMID- 17913322 TI - Lithium effects on brain glutamatergic and GABAergic systems of healthy volunteers as measured by proton magnetic resonance spectroscopy. AB - Lithium is a first-line medicinal treatment for acute bipolar disorder and is also used prophylactically in manic depressive illnesses; however, its mechanism of action is still largely unknown. Animal and human studies have suggested that lithium modulates glutamatergic and GABAergic neurotransmissions. The aim of this study is to investigate the effects of lithium on brain glutamate (Glu), glutamine (Gln), and gamma-aminobutyric acid (GABA) levels in healthy individuals using proton magnetic resonance spectroscopy (1H-MRS). In vivo 3 Tesla 1H-MRS was performed on the anterior cingulate cortex and bilateral basal ganglia initially and after two weeks of lithium administration on 8 healthy male subjects who had a mean age of 34.9 years. After two weeks of lithium administration, Gln significantly decreased in the left basal ganglia and showed a decreasing trend in the right basal ganglia. Additionally, Glu+Gln (Glx) significantly decreased in the right basal ganglia and showed a decreasing trend in the left basal ganglia. Glu did not significantly change in any of the three tested areas, and GABA exhibited no significant change after the lithium administration when measured in the anterior cingulate cortex and left basal ganglia. This study is the first to demonstrate that subchronic lithium treatment decreases Gln and Glx levels in the bilateral basal ganglia of healthy individuals. Our finding might suggest that the decrease of Glx levels is associated with the pharmacological actions of subchronic lithium treatment. PMID- 17913323 TI - ABCB1 (MDR1) gene polymorphisms are associated with the clinical response to paroxetine in patients with major depressive disorder. AB - Variability in antidepressant response is due to genetic and environmental factors. Among genetic factors, the ones controlling for availability of the drug at the target site are interesting candidates. Multidrug resistance 1 (ABCB1, MDR1) gene encodes a blood-brain barrier transporter P-glycoprotein that plays an important role in controlling the passage of substances between the blood and brain. In the present study, we therefore examined the possible association of 3 functional ABCB1 polymorphisms (C3435T: rs1045642, G2677T/A: rs2032582 and C1236T: rs1128503) with response to paroxetine in a Japanese major depression sample followed for 6 weeks. Analysis of covariance at week 6 with baseline scores included in the model as covariate showed significant association of the non-synonymous SNP G2677T/A with treatment response to paroxetine (p=0.011). Furthermore, the wild variants haplotype (3435C-2677G-1236T) resulted associated with poor response (p=0.006). To our best knowledge, this study is the first suggestion of a possible association of ABCB1 variants with SSRIs response. PMID- 17913324 TI - Hypothalamic-pituitary-adrenal axis activity, dehydroepiandrosterone sulphate and their relationships with aggression in early and late alcohol withdrawal. AB - The study aims at investigating the relationship between hypothalamic-pituitary adrenal (HPA) axis alterations and aggression level in alcoholic patients during early and late alcohol withdrawal. Serum levels of basal cortisol and dehydroepiandrosterone sulphate (DHEAS) were measured three times, and cortisol and DHEAS response to dexamethasone twice during the early and late withdrawal periods in alcohol dependent males (n=30) and once in healthy control males (n=20). Abnormal cortisol non-suppression response to dexamethasone in dexamethasone suppression test (DST) was observed in some proportion of the patients in early withdrawal, which normalized in late withdrawal. The study revealed reduced basal DHEAS levels and reduced DHEAS response to dexamethasone in late withdrawal. When the patients were assessed in two separate groups as high- and low-aggressives, in the high-aggression group abnormality in DST was observed during both early and late withdrawal periods, in the low-aggression group it was observed only in early withdrawal. While basal DHEAS levels were low in the high-aggression group only in early withdrawal, it was reduced in the low aggression group during late withdrawal period. Some alterations of the HPA axis during alcohol withdrawal might be associated not only with alcohol use per se but also with aggressivity tendency of alcoholic patients. PMID- 17913326 TI - Mathematical modeling of molar tooth preparations for complete crowns. AB - OBJECTIVES: This study used both cone frustum and right truncated pyramids mathematical methods to evaluate and characterize molar tooth surface area. Previous mathematical modeling studies evaluated only the total surface area. We attempted to analyze both the top surface and lateral surface areas. METHODS: The computations were performed according to the published formulas. The vertical heights of both models were assigned to be 3 and 4mm and the horizontal base width chosen to be 9 and 10mm for both models. Three total occlusal convergences: 10 degrees , 15 degrees , and 20 degrees were selected to be the tested parameters. RESULTS: The calculated data revealed that the top surface area represented a relatively large proportion of the total surface area in both models. We also found that the effect of increasing the total occlusal convergence has a much greater effect on the overall top surface area than the lateral surface and total surface areas. CONCLUSIONS: The models previously used to report and compare surface areas of prepared natural teeth were found to be overestimating the overall surface area. More complex configurations including two-plane reductions, anatomy of cusps and retentive features such as slots and grooves need to be included in future modeling protocols. PMID- 17913325 TI - Influence of Er:YAG laser irradiation distance on the bond strength of a restorative system to enamel by D.T. Chimello-Sousa et al. [J. Dentist. 34 (2006) 245-251]. PMID- 17913327 TI - Effect of various calcium/phosphates ratios of carboxymethylcellulose-based saliva substitutes on mineral loss of bovine enamel in vitro. AB - OBJECTIVES: The present study evaluated the effects of various calcium and phosphate concentrations and ratios of carboxymethylcellulose (CMC)-based solutions on the mineral loss of predemineralised bovine enamel in vitro. METHODS: Bovine enamel specimens were prepared, polished and partly covered with nail varnish, thus serving as control of sound enamel. After demineralisation (37 degrees C; pH 5.0; 14 days) the specimens were exposed to CMC-based solutions (20g/l) with various saturations with respect to apatites containing 0.1mM NaF, CaCl2 (0-32 mM) and KH2PO4 (0-52 mM) at two different pH values (5.5 or 6.5). A fluoride-free solution served as control, and four commercially available products were tested as well. The differences in mineral loss (DeltaDeltaZ) between the values prior to (DeltaZ Demin) and after storage (DeltaZ Effect) in the various solutions were evaluated from microradiographs of thin sections (100microm). RESULTS: The general linear model revealed a significant dependency for DeltaDeltaZ on 'calcium' (p<0.001), 'phosphate' (p=0.023), 'fluoride' (p=0.002) and 'pH' (p<0.001). With increasing calcium and phosphate concentrations an increase in DeltaDeltaZ could be observed up to the solution containing the third highest saturation with respect to octacalciumphosphate (3.2), showing a significant remineralisation (p<0.05; t-test). The commercially available products as well as the control groups revealed significantly reduced DeltaDeltaZ values compared to this group (p<0.01; Bonferroni). CONCLUSIONS: A saturation with respect to octacalciumphosphate of 3.2 and a pH of 6.5 enables CMC-based solutions to remineralise bovine enamel in vitro. PMID- 17913329 TI - Diurnal blood pressure variation in the evaluation of early onset severe pre eclampsia. AB - OBJECTIVE: To study the association between diurnal variation in blood pressure, the mean daily blood pressure and various complications of pregnancy in patients presenting with severe pre-eclampsia before 34 weeks' gestation. STUDY DESIGN: Forty-four women presenting to a tertiary hospital in South Africa with severe pre-eclampsia between 28 and 34 weeks' gestation were managed expectantly for at least 8 days. We measured maternal blood pressure every 30 min with the pregnancy validated Spacelabs 90209 automated blood pressure monitor for 24h periods on alternative days. The mean 24-h diastolic blood pressure measurement, the mean diastolic blood pressure for daytime and nighttime, the day-night blood pressure difference and the night-day ratio were compared with the occurrence of abruptio placentae, gestational age at delivery, neonatal intensive care unit admission, birth weight, abnormal umbilical artery Doppler FVW and reason for delivery. RESULTS: One hundred and seventy-six 24-h studies were analyzed. The day-night blood pressure difference decreased with increasing mean diastolic blood pressure (r=-0.323, p<0.0001). A combination of normal mean diastolic blood pressure and normal day-night blood pressure difference was associated with increased gestational age and lower caesarean section rates. CONCLUSION: The combination of mean diastolic blood pressure and day-night blood pressure difference may be a supplementary measurement of disease severity in early onset severe pre-eclampsia and seems to be of prognostic value. PMID- 17913330 TI - Long-term outcome of hysteroscopic endometrial ablation without endometrial preparation. AB - OBJECTIVE: To describe the three-step hysteroscopic endometrial ablation (EA) technique without endometrial preparation, and its long-term outcomes. STUDY DESIGN: Four hundred and thirty-eight premenopausal women with menorrhagia or menometrorrhagia underwent three-step hysteroscopic EA, which consists of rollerball ablation of the fundus and cornual regions, a cutting loop endomyometrial resection of the rest of the cavity, and rollerball redessication of the whole pre-ablated uterine cavity. The main outcome measures were menstrual status, level of satisfaction with the procedure, and the need for repeat ablation or hysterectomy. Questionnaires were completed for 385 women (87.9%) with a mean follow-up of 48.2 months. RESULTS: One hundred and eighty-four responders (47.8%) reported amenorrhea; 177 (46%) had light to normal flow. One patient (0.3%) underwent repeat ablation and 20 (5.2%) underwent hysterectomy: 15 (3.9%) because of endometrial ablation failure and 5 (1.3%) because of indications unrelated to the ablation (three cases of atypical endometrial hyperplasia and two cases of fibroids). Two hundred and ninety-two patients (75.8%) were very satisfied, and 78 (20.3%) satisfied with the results. No major complications occurred and three women (0.8%) became pregnant during the follow up period. CONCLUSIONS: EA is safe and effective means of treating of menorrhagia and menometrorrhagia in premenopausal women, and helps avoid hysterectomy in 95% of patients suffering from heavy bleeding, with or without uterine fibroids. Women should be informed that the procedure is not contraceptive and that pregnancy is possible after treatment. PMID- 17913328 TI - Tumor necrosis factor and stroke: role of the blood-brain barrier. AB - The progression and outcome of stroke is affected by the intricate relationship between the blood-brain barrier (BBB) and tumor necrosis factor alpha (TNFalpha). TNFalpha crosses the intact BBB by a receptor-mediated transport system that is upregulated by CNS trauma and inflammation. In this review, we discuss intracellular trafficking and transcytosis of TNFalpha, regulation of TNFalpha transport after stroke, and the effects of TNFalpha on stroke preconditioning. TNFalpha can activate cytoprotective pathways by pretreatment or persistent exposure to low doses. This explains the paradoxical observation that transport of this proinflammatory cytokine improves the survival and function of hypoxic cells and of mice with stroke. The dual effects of TNFalpha may be related to differential regulation of TNFalpha trafficking downstream to TNFR1 and TNFR2 receptors. As we better understand how peripheral TNFalpha affects its own transport and modulates neuroregeneration, we may be in a better position to pharmacologically manipulate its regulatory transport system to treat stroke. PMID- 17913332 TI - Olive production systems on sloping land: prospects and scenarios. AB - The ultimate objective of the EU Olivero project was to improve the quality of life of the rural population and to assure the sustainable use of the natural resources of land and water in the sloping and mountainous olive production systems (SMOPS) areas in Southern Europe. One specific objective was to develop, with end-users, alternative future scenarios for olive orchards in the five Olivero target areas. This paper discusses the development of these scenarios, and their socio-economic and environmental effects. After presenting the different production systems (SMOPS) and their strengths, weaknesses, opportunities and threats, a general overview is given of the medium- and long term prospects. These have been validated by experts from the olive sector and foresee changes towards abandonment, intensification and organic production. On balance, the changes could lead to lower production of some target areas in future. An analysis of major external factors affecting the future development of SMOPS indicates there will be labour shortages and increased wage rates, reduced subsidies and constant or rising olive oil prices. On the basis of these assumptions, four future scenarios are developed for the five target areas, with the help of a Linear Programming simulation model. The results are presented for two target areas. For the Tras-os-Montes target area in Portugal, three of the four tested scenarios point to a high level of abandonment, while in the most positive scenario the areas under semi-intensive low input and organic SMOPS increase. In the Granada and Jaen target area in Spain, all scenarios hint at intensification, and only the orchards on the steepest slopes are likely to be abandoned. The direction and extent of environmental effects (erosion, fire risk, pollution, water use and biodiversity) differ per scenario, as do the extent of cross-compliance and agri-environmental measures. PMID- 17913331 TI - Myosin dynamics on the millisecond time scale. AB - Myosin is a motor protein associating with actin and ATP. It translates along actin filaments against a force by transduction of free energy liberated with ATP hydrolysis. Various myosin crystal structures define time points during ATPase showing the protein undergoes large conformation change during transduction over a cycle with approximately 10 ms periodicity. The protein conformation trajectory between two intermediates in the cycle is surmised by non-equilibrium Monte Carlo simulation utilizing free-energy minimization. The trajectory shows myosin transduction of free energy to mechanical work giving evidence for: (i) a causal relationship between product release and work production in the native isoform that is correctly disrupted in a chemically modified protein, (ii) the molecular basis of ATP-sensitive tryptophan fluorescence enhancement and acrylamide quenching, (iii) an actin-binding site peptide containing the free-energy barrier to ATPase product release defining the rate limiting step and, (iv) a scenario for actin-activation of myosin ATPase. PMID- 17913333 TI - The feasibility of decontamination of reduced saline sediments from copper using the electrokinetic process. AB - The remediation of metal contamination in estuarine and reduced sediments is another challenge for the electrokinetic process. Problems result from the complex chemical composition of saline reduced sediments. In the experiments described here copper was added into saline reduced sediments to simulate the natural situation. Two constant voltages were applied across the electrokinetic cell to investigate the effect of an increase in potential difference on the electrokinetic process. Acetic acid at pH 6 and 4 was also added to the cathode in separate experiments to enhance copper removal. The results from this study showed that in the unenhanced experiments most of copper remained in the soil and was unaffected by the electrokinetic process. At the end of unenhanced electrokinetic experiments, the copper concentration was found only to be slightly changed in the anode region. Up to 21% of the copper was found precipitated near the anode when the applied voltage was 10 V and this increased to 25% when the applied voltage was 7 V. In the enhanced electrokinetic experiments up to 98% of copper was removed from section 1 and precipitated again in sections 2 and 3. This indicates that a significant amount copper removal from the saline sediments may be achieved by the enhanced electrokinetic process. PMID- 17913335 TI - Shock waves activate in vitro cultured progenitors and precursors of cardiac cell lineages from the human heart. AB - Postischemic cardiomyopathy remains one of the disorders in urgent need of effective noninvasive therapy. It is currently accepted that the isolation, expansion and application of resident cardiac stem cells may hold therapeutic promise for the future. Recently, it has been demonstrated that shock waves (SW) could enhance the expression of vascular endothelial growth factor (VEGF) and its receptor, Flt-1. As the development of angiogenic noninvasive therapy is very important for future therapeutic strategies in cardiovascular diseases, we examined in vitro, the effects of SW treatment on adult resident cardiac primitive cells isolated from bioptic fragments of normal human hearts and from explanted pathologic hearts with postischemic cardiomyopathy. This study demonstrates that SW have positive influence on both the proliferation and the differentiation of cardiomyocytes, smooth muscle and endothelial cells precursors, with a more obvious effect being evident in the cells from normal heart than in those taken from pathologic hearts. Our results suggest that SW treatment could inhibit or retard the pathologic remodeling and functional degradation of the heart if applied during the early stages of heart failure. PMID- 17913338 TI - How much are patients interested in erectile dysfunction treatment after radical prostatectomy? PMID- 17913334 TI - Robust principal component analysis and clustering methods for automated classification of tissue response to ARFI excitation. AB - We introduce a new method for automatic classification of acoustic radiation force impulse (ARFI) displacement profiles using what have been termed "robust" methods for principal component analysis (PCA) and clustering. Unlike classical approaches, the robust methods are less sensitive to high variance outlier profiles and require no a priori information regarding expected tissue response to ARFI excitation. We first validate our methods using synthetic data with additive noise and/or outlier curves. Second, the robust techniques are applied to classifying ARFI displacement profiles acquired in an atherosclerotic familial hypercholesterolemic (FH) pig iliac artery in vivo. The in-vivo classification results are compared with parametric ARFI images showing peak induced displacement and time to 67% recovery and to spatially correlated immunohistochemistry. Our results support that robust techniques outperform conventional PCA and clustering approaches to classification when ARFI data are inclusive of low to relatively high noise levels (up to 5 dB average signal-to noise [SNR] to amplitude) but no outliers: for example, 99.53% correct for robust techniques vs. 97.75% correct for the classical approach. The robust techniques also perform better than conventional approaches when ARFI data are inclusive of moderately high noise levels (10 dB average SNR to amplitude) in addition to a high concentration of outlier displacement profiles (10% outlier content): for example, 99.87% correct for robust techniques vs. 33.33% correct for the classical approach. This work suggests that automatic identification of tissue structures exhibiting similar displacement responses to ARFI excitation is possible, even in the context of outlier profiles. Moreover, this work represents an important first step toward automatic correlation of ARFI data to spatially matched immunohistochemistry. PMID- 17913339 TI - Knee joint injury and repair modeled by membrane systems. AB - Tissue reparative processes following tissue injury are modeled by a basic membrane system, dealing only with objects, non-active membranes, and non deterministic evolution rules. At the biological level, tissue repair is regulated by multiple interactions between cells and macromolecules, the latter acting as signals. Such signals modify cell behavior including proliferation, migration, differentiation, and phagocytosis. The signaling components themselves are produced and removed by the resident cell population, and this set of events may provide additional stimuli for altering cell activities. In this paper we have focused on modeling the biology of events following an injury to the knee joint, and have used hyaluronan (a polymer produced by cartilage and synovial cells) as an example for a signaling component in the healing process. The intrinsic non-determinism of the model is a key feature, which allows a mathematical description of the repair responses as well as a possibility for either functional restoration or chronic degeneration, leading to arthritis. PMID- 17913340 TI - LHR splicing variants and gene expression in the marmoset monkey. AB - In the marmoset monkey, the LHR type II, lacking exon 10, is the native receptor type. We characterised the LHR splicing pattern in marmoset testes and the adrenals during puberty and in pre- and postpubertal ovaries and quantified mRNA LHR expression in the testis. We detected 11 LHR splicing variants expressed at similar levels and generated by exon skipping and/or usage of cryptic splice sites. No preferred splicing variant during pubertal maturation was observed in both sexes. Testicular and adrenal LHR expression levels did not significantly change with age. However, a significant increase during pubertal maturation for the serum testosterone/LHR ratio indicated that testosterone secretion increases in the presence of constant LHR mRNA expression levels. We conclude that LHR splicing in the marmoset displays a homogenous pattern and that the main function of the LHR is established in the testis, reaching its highest efficiency during pubertal maturation. PMID- 17913342 TI - Somatostatin/somatostatin receptor signalling: phosphotyrosine phosphatases. AB - Activation of phosphotyrosine phosphatases (PTPs) by somatostatin receptor (SSTR) represents one of the main intracellular mechanisms involved in the antiproliferative effect of somatostatin (SST) and analogues. Since their molecular cloning, the role of PTPs is emerging as a major regulator of different cell functions including cell proliferation, differentiation, cell to cell interactions, cell matrix adhesion and cell migration. It was demonstrated that PTPs possess high substrate specificity and their activity is tightly regulated. Importantly, different G protein-coupled receptors transduce their biological activities through PTPs. PTPs were identified as down-stream effectors of SSTRs to transduce antiproliferative signals, and so far, three family members (SHP-1, SHP-2 and DEP-1/PTPeta) have been identified as selective SSTR intracellular effectors. Here, the molecular mechanisms leading SSTRs to regulate PTP activity are discussed, focusing on recent data showing a close interplay between PTPs and tyrosine kinases to transduce tumoral cell growth arrest following SST analogs administration. PMID- 17913341 TI - Genetic abnormalities of somatostatin receptors in pituitary tumors. AB - Somatostatin exerts antisecretive and antiproliferative effects on different endocrine cells by acting through a family of G protein-coupled receptors that includes five subtypes (SST1-5). Normal human pituitary and pituitary adenomas have been shown to express almost all SST subtypes, with the exception of SST4. Consistent with the observation that octreotide and other somatostatin analogs bind to SST2 and SST5 with high affinity, these genes have been screened for quantitative/qualitative abnormalities in tumors removed from patients with poor responsiveness to somatostatin analogs treatment. Data obtained in GH-secreting adenomas suggested that resistance to octreotide was frequently associated with low expression of SST2 mRNA, although other authors failed to confirm this finding. To date, the only mutational change involving SST2 and SST5 is the Arg to Trp substitution in codon 240 of the SST5 gene that was found in one acromegalic patient resistant to octreotide. Similarly, loss of heterozygosis at SST5 gene locus in pituitary adenomas has been described in individual tumors. In recent years, molecular studies investigated the possible association of gene polymorphisms and susceptibility to diseases and/or resistance to drugs. As far as polymorphic variants of SST genes are concerned, a possible role of SST5 C1004T and T-461C alleles in influencing GH and IGF-I levels in patients with acromegaly has been proposed. Nevertheless, polymorphic variants in SST2 and SST5 genes seem to have a minor, if any, role in determining the different responsiveness to somatostatin analogs in patients with acromegaly. PMID- 17913343 TI - Delirium following abrupt discontinuation of fluoxetine. AB - Sudden discontinuation of serotonin reuptake inhibitors (SRI) can lead to a number of psychological (e.g., nervousness, anxiety, crying spells, psychomotor agitation, irritability, depersonalization, decreased mood, memory disturbances, confusion, decreased concentration, and/or slowed thinking) and somatic (e.g., nausea, dizziness, headache) symptoms. Recent studies have shown that withdrawal symptoms are common with paroxetine, venlafaxine and fluvoxamine, but relatively rare and mild with fluoxetine cessation, likely as a result of its longer half life. We report an unusual case of a patient who developed delirium after abrupt discontinuation of fluoxetine. PMID- 17913345 TI - Intracranial falx chondroma: literature review and a case report. AB - Intracranial chondromas are benign tumors that represent approximately 0.5% of all intracranial tumors. They usually occur at the base of the skull but on rare occasions might present as intra-parenchymal or intra-ventricular space-occupying lesions, most likely originating from heterotopic chondrocytes or metaplastic fibroblasts of the falx, the convexity dura, or the ventricular ependyma. Chondromas are slow-growing tumors, which remain clinically silent for a prolonged period of time in the majority of cases. Their clinical presentation is non-specific and their radiographic appearance usually does not differentiate them from their more common counterparts, such as meningiomas and glial tumors. As a result, their diagnosis has remained a histopathologic one. In our current study, we present a case of a falcine intracranial chondroma. This case provided the opportunity to extensively review the pertinent literature. PMID- 17913344 TI - Neuromyelitis optica in a patient with an early onset demyelinating episode: clinical and autoantibody findings. AB - Recent clinical and laboratory findings have substantially advanced our understanding of neuromyelitis optica (NMO) as a humorally mediated, autoimmune disorder. We report on a patient who suffered a first episode of transverse myelitis at the age of 6 months following diphtheria-pertussis-tetanus (DPT) vaccination which had therefore been considered suggestive of acute disseminated encephalomyelitis (ADEM). Fifteen years later, the further disease course revealed typical NMO meeting all diagnostic criteria. This development points to a broad clinical and temporal heterogeneity of NMO, with ADEM probably occurring in the context of a shared autoimmune diathesis. Despite therapy response following B-cell depletion by rituximab, positive NMO-IgG autoantibody status remained unchanged, whereas direct testing for anti-aquaporin-4 (AQP-4) antibodies was negative throughout. Our findings challenge the pathogenic relevance of NMO-IgG and indicate a varying diagnostic value of testing for NMO IgG and AQP-4-autoantibodies. PMID- 17913346 TI - Relationship between visual hallucinations and REM sleep behavior disorder in patients with Parkinson's disease. AB - OBJECTIVES: REM sleep behavior disorder (RBD) has been documented to precede or to co-occur with Parkinson's disease (PD). Parkinson's disease is one of the most common neurological conditions associated with visual hallucinations. Cognitive dysfunction is present in PD, even at the early stages of these diseases. In this study we aimed to investigate the relationship between visual hallucinations and RBD in patients with idiopathic Parkinson's disease (IPD). Additionally, we evaluated the association of the cognition and the pattern of cognitive impairment with VHs and RBD, effects of factors like duration and severity of the disease and duration of levodopa usage. PATIENTS AND METHODS: Seventy-nine patients, diagnosed as PD, were included the study and then, patients were divided into four groups; with RBD and VHs (group 1), with RBD but no VHs (group 2), with VHs but no RBD (group 3), without RBD and VHs (group 4). We compared each group with the others according to demographic characteristics and neuropsychological test scores. RESULTS: Of all patients, in 46% (n=36) RBD and in 48% (n=38) VHs were observed. Our study established VHs in 58% of patients with RBD, and RBD in 55% of patients with VHs. However, due to a 40% incidence of VHs in patients without RBD, RBD and VHs were not found to be correlated. All of the neuropsychometric test scores did not reveal significant difference between groups. CONCLUSION: Although it seems like there is a small association between RBD and VHs in our patients, it was not significant. Group 1 presented with significantly worse scores in UPDRS total scores and I, II subscores. PMID- 17913347 TI - Bilateral cerebellar infarction in the medial branches of posterior inferior cerebellar arterial territory--using endoscopic third ventriculostomy to relieve acute hydrocephalus. AB - Simultaneous bilateral cerebellar infarctions in posterior inferior cerebellar arterial (PICA) territory, without brain stem involvement are rare. We herein report a 51-year-old man developed sudden dizziness, nausea and vomiting. CT revealed hypodense bilateral lesions over the cerebellum corresponding to the medial PICA (mPICA) branch territory. His mental state deteriorated 2 days after onset; repeated CT showed severe third and lateral ventricular dilation. Endoscopic third ventriculostomy (ETV) was done to relieve the acute obstructive hydrocephalus. The patient was later discharged with only mild residual ataxia. Compared with conventional surgical treatments (external ventricular drainage, craniectomy and cerebellectomy), ETV has several advantages, including less risk and minimal invasiveness. However, further study is needed on its safety and efficacy under such circumstances. PMID- 17913348 TI - The removal of the indigo carmine dye from aqueous solutions using cross-linked chitosan: evaluation of adsorption thermodynamics using a full factorial design. AB - A 2(3) factorial design was employed to evaluate the quantitative removal of the indigo carmine (IC) dye from aqueous solutions on glutaraldehyde cross-linked chitosan. The variables were chitosan masses of 100 and 300 mg, IC concentrations of 2.0 and 5.0 x 10(-5) mol L(-1) and temperatures of 25 and 35 degrees C. The quantitative and energetic adsorption parameters were analyzed statistically using modeling with bilinear equations. The results indicated that increasing the chitosan mass from 100 to 300 mg decreases the IC adsorption/mass ratio (mol g( 1)) whereas a temperature increase of 25-35 degrees C increases it. The principal effect of the IC concentration did not show statistical significance. The factorial experiments demonstrate the existence of a significant antagonistic interaction effect between the chitosan mass and temperature. The adsorption thermodynamic parameters, namely Delta(ads)H, Delta(ads)G, and Delta(ads)S, were determined for all the factorial design results. Endothermic values were found in relation to the Delta(ads)H. The positive Delta(ads)S values indicate that entropy is a driving force for adsorption. The Delta(ads)G values are also significantly affected by important antagonistic and synergistic effects involving all principal and interactive factors. It is concluded that the thermodynamical spontaneity of the IC adsorption parameters are greatly influenced by the interactive factors and not by the temperature changes alone. PMID- 17913349 TI - Development of empirical models for performance evaluation of UASB reactors treating poultry manure wastewater under different operational conditions. AB - A nonlinear modeling study was carried out to evaluate the performance of UASB reactors treating poultry manure wastewater under different organic and hydraulic loading conditions. Two identical pilot scale up-flow anaerobic sludge blanket (UASB) reactors (15.7 L) were run at mesophilic conditions (30-35 degrees C) in a temperature-controlled environment with three hydraulic retention times (theta) of 15.7, 12 and 8.0 days. Imposed volumetric organic loading rates (L(V)) ranged from 0.65 to 4.257 kg COD/(m(3) day). The pH of the feed varied between 6.68 and 7.82. The hydraulic loading rates (L(H)) were controlled between 0.105 and 0.21 m(3)/(m(2)day). The daily biogas production rates ranged between 4.2 and 29.4 L/day. High volumetric COD removal rates (R(V)) ranging from 0.546 to 3.779 kg COD(removed)/(m(3)day) were achieved. On the basis of experimental results, two empirical models having a satisfactory correlation coefficient of about 0.9954 and 0.9416 were developed to predict daily biogas production (Q(g)) and effluent COD concentration (S(e)), respectively. Findings of this modeling study showed that optimal COD removals ranging from 86.3% to 90.6% were predicted with HRTs of 7.9, 9.5, 11.2, 12.6, 13.7 and 14.3 days, and L(V) of 1.27, 1.58, 1.78, 1.99, 2.20 and 2.45 kg COD/(m(3)day) for the corresponding influent substrate concentrations (S(i)) of 10,000, 15,000, 20,000, 25,000, 30,000 and 35,000 mg/L, respectively. PMID- 17913350 TI - Sulfur mustard destruction using ozone, UV, hydrogen peroxide and their combination. AB - Numerous methods are used for destruction of sulfur mustard. Oxidation is one of those methods. There have been only limited data concerning application of the advanced oxidation technologies (AOTs) for mustard destruction available before. In this study sulfur mustard oxidation rate depending on kind of the oxidative system and process parameters used was assessed using selected AOT. The following were selected for mustard oxidation: ozone (O(3)), UV light (UV), hydrogen peroxide (H(2)O(2)); double systems: UV/O(3), UV/H(2)O(2), and O(3)/H(2)O(2); a triple system: O(3)/H(2)O(2)/UV and Fenton reaction. Effectiveness of the selected AOT methods has been evaluated and the most suitable one for mustard destruction was chosen. Using ozone in various combinations with hydrogen peroxide and UV radiation mustard can be destroyed much quicker comparing to the classical oxidizers. Fast mustard oxidation (a few minutes) occurred in those systems where ozone alone was used, or in the following combinations: O(3)/H(2)O(2), O(3)/UV and O(3)/H(2)O(2)/UV. When those advanced oxidation technologies are used, mustard becomes destroyed mainly in course of the direct oxidation with ozone, and reactions of mustard with radicals formed due to ozone action play a secondary role. Rate of sulfur mustard oxidation in the above mentioned ozone-containing oxidative systems decreases with pH value increasing from 2 to 12. Only when pH value of reaction solutions is close to pH 5, mustard oxidation rate is minimal, probably due to "disappearance" of radicals participating in oxidation in this pH. Sulfur mustard can be most effectively destroyed using just ozone in pH 7. In that case mustard destruction rate is only slightly lower than the rate achieved in optimal conditions, and the system is the simplest. PMID- 17913351 TI - Exploring multi-metal biosorption by indigenous metal-hyperresistant Enterobacter sp. J1 using experimental design methodologies. AB - A novel experimental design, combining mixture design and response surface methodology (RSM), was developed to investigate the competitive adsorption behavior of lead, copper and cadmium by an indigenous isolate Enterobacter sp. J1 able to tolerate high concentrations of a variety of heavy metals. Using the proposed combinative experimental design, two different experiment designs in a ternary metal biosorption system can be integrated to a succinct experiment and the number of experimental trials was markedly reduced from 38 to 26 by reusing the mutual experimental data. Triangular contour diagrams and triangular three dimensional surface plots were generated to describe the ternary metal biosorption equilibrium data in mixture design systems. The results show that the preference of metal sorption of Enterobacter sp. J1 decreased in the order of Pb(2+)>Cu(2+)>Cd(2+). The presence of other metals resulted in a competitive effect. The influence of the other two metals in ternary metal biosorption system can be easily determined by comparing the stray distance from the single metal biosorption. The behavior of competitive biosorption was successfully described and predicted using a combined Langmuir-Freundlich model along with new three dimensional contour-surface plots. PMID- 17913352 TI - Immobilization and leaching characteristics of arsenic from cement and/or lime solidified/stabilized spent adsorbent containing arsenic. AB - Solidification/stabilization (S/S) of hazardous iron oxide coated cement (IOCC) spent adsorbent containing arsenic (As(III)) was investigated in the present study. Cement and lime-based S/S effectiveness was evaluated by performing semi dynamic leach tests. The S/S effectiveness was evaluated by measuring effective diffusion coefficients (D(e)) and leachability indices (LX). It was found that though cement or lime alone were efficient in preventing arsenic leaching (D(e) being in range of 10(-10) to 10(-12) for all the matrices) from the solidified matrices, the best combination for arsenic containment in the matrix was obtained when a mixture of cement and lime was used. The LX values for all the matrices were higher than 10, suggesting that the S/S treated arsenic sludge are acceptable for "controlled utilization". Calcite formation along with precipitation and conversion into non-soluble forms (calcium arsenite, calcium hydrogen arsenate hydrates, calcium hydrogen arsenates, etc.) were found to be the responsible mechanism for low leaching of arsenic from the solidified/stabilized samples. A linear relationship between cumulative fraction (CFR) of arsenic leached and square root of leach time (R(2) ranging from 0.90 to 0.94) suggested that the diffusion is the responsible mechanism for arsenic leaching. Thus, cement and lime show effective containment of the As(III) within the matrix thus indicating S/S by cement and lime, which is also a low-cost option, as a suitable management option for the toxic As(III) sludge. PMID- 17913353 TI - Sulfide removal in petroleum refinery wastewater by chemical precipitation. AB - Sulfide removal by chemical precipitation from petroleum refinery wastewater was investigated. The wastewater samples were taken from the flocculation pond influent of TUPRAS Kirikkale Middle Anatolia Petroleum Refinery Wastewater Treatment Plant (WWTP) and physicochemical treatments using conventional coagulants which were partial precipitant [FeCl(3) . 6H(2)O and FeSO(4) . 7H(2)O] and coagulant-aids [Ca(OH)(2) and CaCO(3)] were applied to both raw and sulfide added wastewater. Sulfide and chemical oxygen demand (COD) removal efficiencies of Fe(3+) ions alone for sulfide added wastewaters having different pH values varied between 62-95 and 45-75%, respectively. In addition, removal efficiencies of sulfide (96-99%) and COD (50-80%) were obtained by using Fe(2+) ions together with Ca(OH)(2) as precipitant-aid under the same conditions. In experiments performed with raw wastewater which had different pH values, COD removal efficiencies of Fe(3+) and Fe(2+) ions together with Ca(OH)(2), were 50-80 and 32 50%, respectively. PMID- 17913354 TI - Competition of Reactive red 4, Reactive orange 16 and Basic blue 3 during biosorption of Reactive blue 4 by polysulfone-immobilized Corynebacterium glutamicum. AB - Competition of Reactive red 4 (RR4), Reactive orange 16 (RO16) and Basic blue 3 (BB3) during biosorption of Reactive blue 4 (RB4) by polysulfone-immobilized protonated Corynebacterium glutamicum (PIPC) was investigated in batch and column mode of operations. Through potentiometric titrations, and with the aid of proton binding model, carboxyl, phosphonate and amine were identified as functional groups of PIPC, with apparent pK(a) values of 3.47+/-0.05, 7.08+/-0.07 and 9.90+/ 0.05 mmol/g, respectively. Since reactive dyes release dye anions (ROSO(3)(-)) in solutions, the positively charged amine groups were responsible for biosorption. PIPC favored biosorption at pH 3 when RB4 was studied/used as single-solute; while the presence of RR4 and RO16 severely affected the RB4 biosorption. When present as a single-solute, PIPC recorded 184.5mg RB4/g; while PIPC exhibited 126.9, 120.9 and 169.6 mg RB4/g in the presence of RR4, RO16 and BB3, respectively. In general, the accessibility of amine group depends on the molecular size, number of sulfonate groups and reactivity of each reactive dye. Single and multicomponent Freundlich equations successfully described the biosorption isotherms. With 0.1M NaOH, it is possible to reuse PIPC for RB4 biosorption in 10 repeated cycles. Column experiments in an up-flow packed column coincided with batch results, that is PIPC showed strong preference towards highly reactive and relatively small RB4 anions; however, the presence of competing dyes hinder the RB4 column biosorption performance. PMID- 17913355 TI - Photodegradation of 2-mercaptobenzothiazole in the gamma-Fe(2)O(3)/oxalate suspension under UVA light irradiation. AB - The aim of this study is to investigate the effect of various factors on the photodegradation of organic pollutants in natural environment with co-existence of iron oxides and oxalic acid. 2-Mercaptobenzothiazole (MBT) was selected as a model pollutant, while gamma-Fe(2)O(3) was selected as iron oxide. The crystal structure and morphology of the prepared gamma-Fe(2)O(3) was determined by X-ray diffractograms (XRD) and scanning electron microscopy (SEM), respectively. The specific surface area was 14.36 m(2)/g by Brunauer-Emmett-Teller (BET) method. The adsorption behavior of gamma-Fe(2)O(3) was evaluated by Langmuir model. The effect of the dosage of iron oxide, initial concentration of oxalic acid (C(ox)(0)), initial pH value, the light intensity and additional transition metal cations on MBT photodegradation was investigated in the gamma-Fe(2)O(3)/oxalate suspension under UVA light irradiation. The optimal gamma-Fe(2)O(3) dosage was 0.4 g/L and the optimal C(ox)(0) was 0.8 mM with the UVA light intensity of 1800 mW/cm(2). And the optimal dosage of gamma-Fe(2)O(3) and C(ox)(0) for MBT degradation also depended strongly on the light intensity. The optimal gamma Fe(2)O(3) dosage was 0.1, 0.25 and 0.4 g/L, and the optimal C(ox)(0) was 1.0, 0.8, and 0.8 mM with the light intensity of 600, 1200 and 1800 mW/cm(2), respectively. The optimal initial pH value was at 3.0. The additional transition metal cations including Cu(2+), Ni(2+) or Mn(2+) could significantly accelerate MBT degradation. This investigation will give a new insight to understanding the MBT photodegradation in natural environment. PMID- 17913356 TI - Modeling small-scale spills of aqueous solutions in the indoor environment. AB - A mass transfer model is proposed to estimate the rates of chemical emissions from aqueous solutions spilled on hard surfaces inside buildings. The model is presented in two forms: a set of four ordinary differential equations and a simplified exact solution. The latter can be implemented in a spreadsheet. User input includes ten parameters, which represent either the properties of the source or those of the building. All of them can be readily obtained. The proposed model is tested against and in good agreement with the measurements of simulated spill events in a room-sized environmental chamber. This model can be used by emergency response planners to estimate the time history of contaminant concentrations in indoor air. PMID- 17913357 TI - Effective utilization of waste ash from MSW and coal co-combustion power plant: Zeolite synthesis. AB - The solid by-product from power plant fueled with municipal solid waste and coal was used as a raw material to synthesize zeolite by fusion-hydrothermal process in order to effectively use this type of waste material. The effects of treatment conditions, including NaOH/ash ratio, operating temperature and hydrothermal reaction time, were investigated, and the product was applied to simulated wastewater treatment. The optimal conditions for zeolite X synthesis were: NaOH/ash ratio=1.2:1, fusion temperature=550 degrees C, crystallization time=6-10 h and crystallization temperature=90 degrees C. In the synthesis process, it was found that zeolite X tended to transform into zeolite HS when NaOH/ash ratio was 1.8 or higher, crystallization time was 14-18 h, operating temperature was 130 degrees C or higher. The CEC value, BET surface area and pore volume for the synthesized product at optimal conditions were 250 cmol kg(-1), 249 m(2) g(-1) and 0.46 cm(3) g(-1) respectively, higher than coal fly ash based zeolite. Furthermore, when applied to Zn(2+) contaminated wastewater treatment, the synthesized product presented larger adsorption capacity and bond energy than coal fly ash based zeolite, and the adsorption isotherm data could be well described by Langmuir and Freundlich isotherm models. These results demonstrated that the special type of co-combustion ash from power plant is suitable for synthesizing high quality zeolite, and the products are suitable for heavy metal removal from wastewater. PMID- 17913359 TI - Comparison of the properties of glass, glass-ceramic and ceramic materials produced from coal fly ash. AB - Glass, glass-ceramic and ceramic materials were produced from thermal power plant fly ash without any additives. X-ray diffraction (XRD) analysis revealed the amorphous phase of the glass sample. Augite phase was detected in the glass ceramic sample, while the enstatite and mullite phases occurred in the ceramic samples. Scanning electron microscopy (SEM) investigations showed that tiny crystallites homogeneously dispersed in the microstructure of the glass-ceramic sample and elongated crystals formed in the ceramic samples. Density values of the obtained samples are comparable to those of the commercially produced glass, glass-ceramic and ceramic samples. Toxicity characteristic leaching procedure (TCLP) results indicated that the produced samples could be taken as non hazardous materials. Produced samples showed high resistance to alkali solutions in contrast to acidic solutions. Microstructural, physical, chemical and mechanical properties of the produced glass-ceramic samples are better than those of the produced glass and ceramic samples. PMID- 17913358 TI - Preparation, characterization, non-isothermal reaction kinetics, thermodynamic properties, and safety performances of high nitrogen compound: hydrazine 3-nitro 1,2,4-triazol-5-one complex. AB - A new high nitrogen compound hydrazine 3-nitro-1,2,4-triazol-5-one complex (HNTO) was prepared by the reaction of 3-nitro-1,2,4-triazol-5-one with hydrazine hydrate, and its structure was characterized by means of organic elemental analyzer, FT-IR, XRD, (13)C NMR and (15)N NMR. The non-isothermal reaction kinetics of the main exothermic decomposition reaction of HNTO was investigated by means of DSC. The thermodynamic properties of HNTO were calculated. The results showed that the formation of HNTO is achieved by proton transfer of N(4) atom, and it makes a higher nitrogen content and lower acidity. The reaction mechanism of HNTO is classified as nucleation and growth, and the mechanism function is Avramo-Erofeev equation with n=2/5. The kinetic parameters of the reaction are E(a)=195.29 kJ mol(-1), lg(A (s(-1)))=19.37, respectively. The kinetic equation can be expressed as: d(alpha)/d(t) = 10(18.97)(1 - alpha)[-ln(1 alpha)](3/5) e(-2.35 x 10(4)/T). The safety performances of HNTO were carried out. The critical temperature of thermal explosion are 464.26 and 474.37 K, the adiabatic time-to-explosion is 262s, the impact sensitivity H(50)=45.7 cm, the friction sensitivity P=20% and the electrostatic spark sensitivity E(50)>5.4J (no ignition). It shows that HNTO has an insensitive nature as RDX and NTO, etc. PMID- 17913360 TI - Coeruleospinal inhibition of visceral nociceptive processing in the rat spinal cord. AB - Visceral nociceptive information is transmitted in two different areas of the spinal cord gray matter, the dorsal horn and the area near the central canal. The present study was designed to examine whether visceral nociceptive transmission in the two different areas is under the control of the centrifugal pathways from the locus coeruleus/subcoeruleus (LC/SC). Extracellular recordings were made from the L(6)-S(2) segmental level using a carbon filament glass microelectrode (4-6 MOmega). Colorectal distentions (80 mmHg) were produced by inflating a balloon inside the descending colon and rectum. In both dorsal horn and deep area neurons, responses to colorectal distention were inhibited during electrical stimulation (30, 50 and 70 microA, 100 Hz, 0.1 ms pulses) of the LC/SC. It is well known that spinothalamic tract (STT) neurons excited by visceral nociceptive stimuli are located in the dorsal horn and that postsynaptic dorsal column (PSDC) neurons which conduct visceral nociceptive signals in the dorsal column (DC) are located near the central canal of the spinal cord. The present study, therefore, suggests that the descending LC/SC system can inhibit visceral nociceptive signals ascending through the STT and the DC pathways. PMID- 17913361 TI - Cell imaging by coherent backscattering microscopy using frequency-shifted optical feedback in a microchip laser. AB - In this work, we present a new development of the laser optical feedback imaging technique for imaging biological structures with a high resolution. The first results obtained on human red blood cells and mice cerebral and muscular tissues slices are shown. The performances of the system and its future developments are also discussed. PMID- 17913362 TI - Scanning tunneling microscopy of Fe- and O-sublattices on Fe3O4(100). AB - We present scanning tunneling microscopy of an octahedral (B) plane terminated (square root of 2 x square root of 2) R45 degrees-reconstructed surface of a natural magnetite (100) crystal. Implementing a W-tip we achieve the same resolution on Fe rows as was reported in the past either with the use of antiferromagnetic tips or on magnetite (Fe3O4) films. We show images of Fe or O sublattices of Fe3O4 with atomic resolution. PMID- 17913363 TI - Changes to plasminogen activators and matrix metalloproteinase-9 in dogs with toxocarosis. AB - The relationship between proteolytic enzymes and hematological response to infection was studied in five 1-month-old dogs inoculated experimentally with 2000 eggs of Toxocara canis. Moderate leukocytosis and marked eosinophilia occurred 14 days post-infection with T. canis. Plasminogen activators (PAs) and matrix metalloproteinase-9 (MMP-9) activity in serum was significantly different in dogs infected with T. canis, compared with controls. Urokinase-type PA (uPA) activity was positively correlated with eosinophilia, and tissue-type PA (tPA) and MMP-9 activity was negatively correlated with eosinophilia. However, there was no correlation between inflammation and MMP-2. The use of uPA, tPA or MMP-9 proteolytic enzymes as laboratory reference markers for toxocarosis requires further study. PMID- 17913364 TI - Pharmacokinetics of metaflumizone in the plasma and hair of cats following topical application. AB - Controlled laboratory studies have shown that a novel spot-on formulation containing 20% (w/v) metaflumizone (ProMeris for Cats, Fort Dodge Animal Health, Overland Park, KS) is effective for the treatment and control of fleas on cats. Two studies were conducted to determine the distribution of metaflumizone in the plasma and hair of cats following treatment at the minimum recommended dose of 40mg/kg. Six purpose-bred cats, three males and three females, were used in each study. Plasma or hair samples were collected from each cat just prior to dosing and periodically through 56 days after treatment. Samples were analyzed by HPLC methods validated for the determination of metaflumizone. Metaflumizone concentrations in plasma were below the method limit of quantification (<50ng/ml) in all samples but one, and were frequently not detectable (<1.1ng/ml). Plasma collected 3 days post-treatment from one cat had a metaflumizone concentration of 57.8ng/ml. The frequency of measurable levels of metaflumizone in the plasma was too low to allow the calculation of pharmacokinetic parameters. Analysis of hair samples indicated that metaflumizone was widely distributed in the hair coat of the cat within 1 day after administration, reaching maximum concentrations within 1 or 2 days post-treatment. Low but quantifiable levels were still present at the end of the 56-day study. Data from the present studies indicate that the ectoparasitic activity is due to exposure of the parasites to metaflumizone on the surface of the host (skin and hair), not to exposure via the circulatory system of the host. PMID- 17913365 TI - The molecular characterisation of Giardia from dogs in southern Germany. AB - Faecal samples were obtained randomly from asymptomatic dogs visiting veterinary clinics (kept individually) and sanctuaries (kept in groups) in southern Germany. Of 60 Giardia positive samples, 55 samples successfully amplified at the 18S rDNA locus. In both dogs kept individually and dogs kept in groups, assemblage A was most prevalent (overall, n=33, 60%), followed by mixed infections with assemblages A&C (overall, n=15, 27.3%), while assemblages C&D alone were less often detected (overall, n=5, 9.1% and n=2, 3.6%). The occurrence of zoonotic genotypes was more prevalent in individual than in group dogs. Genotyping at the gdh locus confirmed the 18S results and clustered the assemblage A isolates into A-I. This is the first large-scale urban survey in Central Europe, which has not only confirmed the high prevalence of Giardia in asymptomatic domestic dogs, but has also shown that the zoonotic assemblage A occurs commonly in domestic dogs living in urban environments, and more frequently than the dog-specific assemblage. Although it was not possible to obtain samples from pet owners in this study, the results demonstrate that a significant proportion of dogs in urban areas of southern Germany harbour zoonotic Giardia, and should thus be considered a potential reservoir for infection in humans. PMID- 17913366 TI - Lipid remodeling of GPI-anchored proteins and its function. AB - Many proteins are attached to the cell surface via a conserved post-translational modification, the glycosylphosphatidylinositol (GPI) anchor. GPI-anchored proteins are functionally diverse, but one of their most striking features is their association with lipid microdomains, which consist mainly of sphingolipids and sterols. GPI-anchored proteins modulate various biological functions when they are incorporated into these specialized domains. The biosynthesis of GPI and its attachment to proteins occurs in the endoplasmic reticulum. The lipid moieties of GPI-anchored proteins are further modified during their transport to the cell surface, and these remodeling processes are essential for the association of proteins with lipid microdomains. Recently, several genes required for GPI lipid remodeling have been identified in yeast and mammalian cells. In this review, we describe the pathways for lipid remodeling of GPI-anchored proteins in yeast and mammalian cells, and discuss how lipid remodeling affects the association of GPI-anchored proteins with microdomains in cellular events. PMID- 17913367 TI - Using giant unilamellar lipid vesicle micro-patterns as ultrasmall reaction containers to observe reversible ATP synthesis/hydrolysis of F0F1-ATPase directly. AB - F(0)F(1)-ATPase within chromatophores, which was labeled with pH-sensitive quantum dots, was encapsulated in large unilamellar lipid vesicles (LUVs) through reverse-phase evaporation. Then a microarray of chromatophore-containing LUVs was created using a micro-contact printing (mu-CP) technique. Through controlled dehydration-rehydration of the lipid patterns, a microarray of single chromatophore-containing giant unilamellar lipid vesicles (GUVs) was formed with desired size and uniform shape. The reversible ATP synthesis/hydrolysis of F(0)F(1)-ATPase in GUVs was directly observed by fluorescence microscopy through the fluorescence intensity increase/decrease in the pH-sensitive quantum dots labeled on the outer surface of the chromatophore. To the best of our knowledge, this is the first direct observation of the reversible behavior of F(0)F(1) ATPase at the bulk scale. PMID- 17913368 TI - Burns. PMID- 17913369 TI - Persistent vegetative state in electrical injuries: a 10-year review. AB - We reviewed 148 cases of electrical injury admitted to our burn centre. The incidence of persistent vegetative state was 3% (n=5), higher in the low-voltage (6.7%) than in high-voltage group (1.2%). At the time of trauma, 44% (n=65) lost consciousness and 50% of these (n=32) received cardiopulmonary resuscitation on arrival at hospital. Of these, 50% recovered (n=16), 22% became comatose (n=7) and 28% (n=9) died. Of the seven comatose patients, five did not show brain oedema but remained in a persistent vegetative state; this state was more common with low-voltage electrical injuries. The public should be warned of this effect of low-voltage trauma. PMID- 17913370 TI - Impairment of the host's antibacterial resistance by norepinephrine activated neutrophils. AB - The susceptibility of mice to infectious complications is dramatically increased in an accompaniment with systemic inflammatory response syndrome (SIRS). Polymorphonuclear neutrophils with immunosuppressive ability (PMN-II) that appear in response to SIRS have been classified as one of the cells responsible for the increased susceptibility of mice with SIRS (SIRS mice) to sepsis induced by cecal ligation and puncture (CLP). Since a high level of norepinephrine (NE) is demonstrated in the plasma of SIRS mice, in the present study, the role of NE on the appearance of PMN-II in SIRS mice was studied. Similar to SIRS mice, normal mice became susceptible to CLP-induced infectious complications after inoculation with NE-treated PMN. CCL2 and IL-10 (biomarkers for PMN-II) were equally produced by PMN-II prepared from SIRS mice and NE-treated PMN. However, CCL3 and IL-12 (biomarkers for immunostimulatory PMN, PMN-I) were not detected in culture fluids from either PMN preparation. These results indicate that NE mass-produced in association with SIRS development plays a role on the generation of PMN-II and the appearing PMN-II are responsible, in part, for increased susceptibility of SIRS mice to CLP-induced infectious complications. PMID- 17913371 TI - Dermabrasion is not a treatment for burns. PMID- 17913372 TI - A three-decade review of thermal injuries among the elderly at a regional burn centre. AB - AIM: To review trends in incidence and treatment of thermal injuries among the elderly. METHOD: A 3-year retrospective review of medical records of people aged 65 years and older admitted to our burn centre over July 2003-June 2006. RESULTS: Elderly people with burns continued to have significant comorbidities. They were often burned because they were inappropriately trying to live alone. Thus discharge was often complicated. CONCLUSIONS: At our burn centre, survival among elderly people with burns has increased, probably as a result of more sophisticated medical, surgical and nursing care, as well as more extensive rehabilitation. PMID- 17913373 TI - Nine-flap z-plasty: a new modification of five-flap z-plasty. PMID- 17913374 TI - Visceral hyperalgesia induced by neonatal maternal separation is associated with nerve growth factor-mediated central neuronal plasticity in rat spinal cord. AB - Neonatal maternal separation (NMS) has been shown to trigger alterations in neuroendocrine, neurochemical and sensory response to nociceptive stimuli along the brain-gut axis. These alterations may be the result of a cascade of events that are regulated by neurotrophic factors. Nerve growth factor (NGF), a member of the neurotrophin family, is essential for the development and maintenance of sensory neurons and for the formation of central pain circuitry. The present study aimed to investigate whether NMS causes changes in neuronal plasticity and the relationship of these changes in plasticity with the expression of NGF and its high affinity tyrosine kinase receptor A (TrkA) in the lumbosacral spinal cord in adult rats. Male Wistar rat pups were either subjected to 180 min daily of NMS or not handled (NH) for 13 consecutive days. The expression of NGF and TrkA was examined in NH and NMS rats with or without colorectal distention (CRD) as determined by Western blot analysis and immunohistochemistry. The present results of Western blot analysis indicated NMS and CRD have a significant effect on NGF protein level in the lumbosacral spinal cord of rats. Assessments of optical densities revealed that NMS enhanced TrkA-ir fiber densities in laminae I III and laminae V-VI of rats in both conditions with or without CRD. Double immunofluorescence revealed that TrkA co-expressed with calcitonin gene-related peptide (CGRP) in afferent fibers, while no significant difference in terms of the intensity of TrkA-ir in these fibers was found among groups. Quantitative analysis of TrkA-ir neurons indicated a significant interactive effect of NMS and CRD on the mean number of TrkA-ir neurons in laminae V-VI of rats, in which significant difference was found between NMS+CRD and NH+CRD. Double immunofluorescence of TrkA and Fos showed that CRD has a significant effect on TrkA expression in Fos-positive neurons in laminae V-VI and lamina X of rats, while no significant difference was found between NMS+CRD and NH+CRD. These results demonstrate that NMS induced alterations in NGF protein level and TrkA expression in adult rat spinal cord and indicate that NGF is a crucial mediator for the changes in neuronal plasticity that occur in NMS-induced visceral hyperalgesia. PMID- 17913375 TI - L-type calcium channel blockade on haloperidol-induced c-Fos expression in the striatum. AB - Haloperidol-induced c-Fos expression in the lateral part of the neostriatum has been correlated with motor side effects while c-Fos induction in the medial part of the neostriatum and the nucleus accumbens is thought to be associated with the therapeutic effects of the drug. Induction of c-Fos in the striatum by haloperidol involves dopamine D(2) (DA D(2)) receptor antagonism and is dependent on activation of N-methyl-d-aspartate (NMDA) receptors and L-type Ca(2+) channels. In the current study, pretreatment with L-type Ca(2+) channel blockers suppressed haloperidol-induced c-Fos throughout the neostriatum and the nucleus accumbens at 2 h postinjection. However, elevated c-Fos protein expression was observed only in the lateral part of the neostriatum at 5 h postinjection of haloperidol following pretreatment of L-type Ca(2+) channel blocker compared with rats pretreated with vehicle alone. In addition, pretreatment prolonged the duration of haloperidol-induced catalepsy in rats. Infusions of L-type Ca(2+) channel blockers directly into the neostriatum mimicked similar patterns of changes in haloperidol-induced c-Fos expression. Prolonged expression of c-Fos was not observed following coadministration of nifedipine and a dopamine D(1) (DA D(1)) receptor agonist, SKF 81297, but could be mimicked by the DA D(2/3) receptor antagonist raclopride, suggesting that the phenomenon is likely related to DA D(2) receptor antagonism. Moreover, the expression levels of haloperidol induced zif 268 and haloperidol-induced phosphorylated CREB and phosphorylated Elk-1 were also substantially elevated for a prolonged period of time in the lateral, but not the medial part of the neostriatum, following blockade of L-type Ca(2+) channels. Collectively, the results suggest that coadministration of L type Ca(2+) channel blockers affects haloperidol signaling in the lateral part of the neostriatum and may exacerbate the development of acute motor side effects. PMID- 17913377 TI - Factors associated with resilience in healthy adults. AB - Mature defenses comprise one well-validated indicator of resilience. We investigated the relationships of resilience to trauma, attachment, temperament, cortisol, and cognitive performance in adult healthy volunteers. Participants were administered the Defense Style Questionnaire; the Relationship Questionnaire; the Childhood Trauma Questionnaire, and the Tridimensional Personality Questionnaire. Cortisol determinations included 24-h urinary, mean hourly plasma, response to low-dose dexamethasone suppression, and reactivity to the Trier social stress test (TSST). Mathematical performance during the TSST was quantified. Twenty-five women and 29 men participated. Resilience was significantly negatively correlated with childhood interpersonal trauma and with harm avoidance. Resilience was significantly positively correlated with urinary cortisol, secure attachment, reward dependence, and superior performance. In a linear regression analysis, the strongest predictor of resilience was childhood trauma, followed by math performance under stress and harm avoidance. We conclude that in young adults without manifest psychiatric disorder, resilience was associated with developmental, biological, and cognitive measures which merit further investigation. PMID- 17913376 TI - Quantitative ultrastructural differences between local and medial septal GABAergic axon terminals in the rat hippocampus. AB - Functionally distinct subsets of hippocampal inhibitory neurons exhibit large differences in the frequency, pattern and short-term plasticity of GABA release from their terminals. Heterogeneity is also evident in the ultrastructural features of GABAergic axon terminals examined in the electron microscope, but it is not known if or how this corresponds to interneuron subtypes. We investigated the feasibility of separating morphologically distinct clusters of terminal types, using the approach of measuring several ultrastructural parameters of GABAergic terminals in the CA1 area of the rat hippocampus. Septo-hippocampal axon terminals were anterogradely labeled by biotinylated dextran amine and visualized by pre-embedding immunogold staining to delineate one homogeneous terminal population. Long series (100-150) of ultrathin sections were cut from stratum oriens and stratum radiatum of the CA1 area, and GABAergic terminals were identified by post-embedding immunogold staining. Stereologically unbiased samples of the total GABAergic axon terminal population and a random sample of the septal axon terminals were reconstructed in 3D, and several of their parameters were measured (e.g. bouton volume, synapse surface, volume occupied by vesicles, mitochondria volume). Septal terminals demonstrated significantly larger mean values for most parameters than the total population of local GABAergic terminals. There was no significant difference between terminals reconstructed in the basal and apical dendritic regions of pyramidal cells, neither for the septal nor for the local population. Importantly, almost all parameters were highly correlated, precluding the possibility of clustering the local terminals into non-overlapping subsets. Factor and cluster analysis confirmed these findings. Our results suggest that similarly to excitatory terminals, inhibitory terminals follow an "ultrastructural size principle," and that the terminals of different interneuron subtypes cannot be distinguished by ultrastructure alone. PMID- 17913378 TI - Characterizing early cigarette use episodes in novice smokers. AB - Retrospective self-report data indicate that early cigarette use episodes may be important predictors of smoking. Unfortunately, recall of early experiences are confounded with current smoking. The current study is the first to examine early cigarette use episodes (EUEs) prospectively in novice smokers (less than 15 lifetime cigarettes). Smoking amount, context and subjective experiences for up to five of the first cigarette episodes during their first year of college were collected using weekly internet-based questionnaires and structured interviews. Data were obtained on 538 EUEs from 163 students. EUEs generally occurred within a social/party context; over 90% of EUEs occurred when participants were with other people who were smoking and over 65% occurred while participants were drinking alcohol. Subjective effects across episodes were reported as generally mild and factor analysis yielded Positive, Negative and Sensory/Peripheral effects scales. Subjective effects were related to the amount smoked and inhalation, whereas EUE context, including alcohol use and social context, was not. This study demonstrates that it is possible to study EUEs in college students within days or weeks of their occurrence and that most of these occur in social settings with the concurrent use of alcohol. PMID- 17913379 TI - The differential impact of relational health on alcohol consumption and consequences in first year college women. AB - The Relational Health Indices (RHI) is a relatively new measure that assesses the strength of relationships. It has been found that relational health has a protective factor for women, such that it enhances positive experiences and limits negative ones. The current study is the first to use the RHI to examine the effect of relational health on alcohol consumption and alcohol consequences. First year college women were given questionnaires assessing relational health, drinking motives, and alcohol use in their first few months at a mid-sized, private university. Due to the social nature of college settings, it was predicted that relational health would moderate the relationship between motives and alcohol consumption. Further, due to the protective factor of relational health, it was predicted that relational health would attenuate the relationship between drinking and negative consequences. These hypotheses were supported. Relational health, moderated the relationship between both social and coping drinking motives and drinking, such that women with strong relational health towards their peers and community who also had high social and coping motives, drank more than those with weaker relationships. Paradoxically, relational health also moderated the relationship between drinking and consequences such that heavy drinking women with strong relational health experienced fewer negative consequences than women with weaker relational health. Results indicate that although relational health is associated with an increase in alcohol consumption, it may also serve as a protective factor for alcohol-related negative consequences. Future research and interventions may seek to de-link the relational health-drinking connection in the college student environment. PMID- 17913381 TI - Effects of repeated days of smoking cue exposure on urge to smoke and physiological reactivity. AB - The present study investigated the effects of repeated days of laboratory-based smoking cue exposure on subjective and physiologic cue reactivity. Twenty non treatment seeking moderate/heavy smokers completed three laboratory sessions approximately 7 days apart, each following a 10-hour nicotine deprivation period. Cue reactivity procedures consisted of a relaxation trial followed by two trials of in vivo cue exposure. Dependent measures included urge to smoke, a withdrawal questionnaire, mean arterial pressure (MAP), and heart rate (HR). A Condition (relaxation vs. cue exposure) by Day (1, 2, or 3) analysis of variance revealed a significant main effect of Condition (greater urge to smoke after cue exposure) but no significant main or interaction effect for Day. Similarly, MAP and HR change scores following cue exposure did not differ across test days. Cue elicited changes in withdrawal symptoms were only observed on Day 1, but not when the interday interval was covaried. Results suggest that laboratory-based cue elicited changes in urge to smoke, MAP, and HR are stable over three separate days. PMID- 17913380 TI - Does gender moderate associations among impulsivity and health-risk behaviors? AB - The present study explores the relations among gender, impulsivity and three health-risk behaviors relevant to young adults (tobacco use, alcohol problems and gambling problems) in a sample of 197 college-age individuals. We sought to determine whether impulsivity is associated with health-risk behaviors in the same ways for men and women. For tobacco use and gambling problems, men were at higher risk than women, and impulsivity was not significantly associated with higher risk. Higher levels of motor impulsivity in men accounted for a significant amount of the gender difference in risk for alcohol problems. That is, impulsivity as measured by the Barratt Impulsiveness Scale (version 11), mediated the association between gender and risk for alcohol problems. For impulsivity as measured by Stop Signal Reaction Time (i.e. response inhibition), gender moderated the association between impulsivity and alcohol problems. Specifically, lower levels of impulsivity were associated with greater risk for alcohol problems in both men and women, but the effect was stronger in men. We speculate that this seemingly paradoxical result might be the result of coping drinking to deal with negative affect associated with behavioral overcontrol. These findings suggest that prevention efforts might well focus on identifying individuals at high risk for alcohol problems, especially males, by assessing response inhibition. PMID- 17913382 TI - Activation of signaling molecules and matrix metalloproteinases in right ventricular myocardium of rats with pulmonary hypertension. AB - Pulmonary hypertension induces right ventricular (RV) overload, which is transmitted to cardiomyocytes via integrins that activate intracellular messengers, including focal adhesion kinase (FAK) and neuronal nitric oxide synthase (NOS1). We investigated whether RV hypertrophy (RVH) and RV failure (RVF) were associated with activation of FAK, NOS1, and matrix metalloproteinases (MMPs). Rats were treated without (RVC) or with a low dose of monocrotaline (30mg/kg) to induce RVH, and with a high dose (80mg/kg) to induce RVF. After approximately 30 days, RV function was determined using a combined pressure conductance catheter. After sacrifice, FAK, NOS1, their phosphorylated forms (FAK P and NOS1-P), MMP-2, and MMP-9 were quantified in RV myocardium by immunohistochemistry. In RVH and RVF, RV weight/ body weight increased by 36% and 109%, whereas RV ejection fraction decreased by 23% and 57% compared to RVC, respectively. FAK-P and FAK-P/FAK were highest in RVH (2.87+/-0.12 and 2.52+/ 0.23 fold compared to RVC, respectively) and slightly elevated in RVF (1.76+/ 0.17 and 1.15+/-0.13 fold compared to RVC, respectively). NOS1-P and NOS1-P/NOS1 were increased in RVH (1.63+/-0.12 and 3.06+/-0.80 fold compared to RVC, respectively) and RVF (2.16+/-0.03 and 3.30+/-0.38 fold compared to RVC, respectively). MMP-2 was highest in RVH and intermediate in RVF (3.50+/-0.12 and 1.84+/-0.22 fold compared to RVC, respectively). MMP-9 was elevated in RVH and RVF (2.39+/-0.35 and 2.92+/-0.68 fold compared to RVC, respectively). Activation of FAK in RVH points to an integrin-dependent hypertrophic response of the myocardium. Activation of NOS1 in failing RV suggests a role of excessive NO in the development of failure and activation of MMPs leading to ventricular remodeling. PMID- 17913383 TI - Stromal tumors of the lower female genital tract: histogenetic, morphological and immunohistochemical similarities with the "benign spindle cell tumors of the mammary stroma". PMID- 17913384 TI - Centrosome abnormalities in non-small cell lung cancer: correlations with DNA aneuploidy and expression of cell cycle regulatory proteins. AB - The aims of this study were to investigate centrosome abnormalities in non-small cell lung cancer (NSCLC), and to assess their relationship with DNA aneuploidy, the expression of the cell cycle-associated proteins, and clinicopathological profiles. Tissue microarrays were constructed from 175 NSCLCs. We analyzed centrosome abnormalities and the expression of p16(INK4a), p53, and pRb using immunohistochemistry. Centrosome abnormalities were noted in 29% of the tumors and were even observed in the normal cells adjacent to the tumor. The frequency of DNA aneuploidy was significantly higher in the tumors containing centrosome abnormalities than in the tumors with a normal centrosome. p16(INK4a) expression and loss of pRb expression, but not p53 expression, were significantly associated with centrosome abnormalities. Clinically, centrosome abnormalities were not found to have any prognostic value for NSCLCs. These results suggest that centrosome abnormalities may be associated with inactive pRb-pathway and contribute to pulmonary carcinogenesis by the level of increasing chromosome instability. PMID- 17913385 TI - Floral leukemic cells transformed from marginal zone lymphoma. AB - There are three clinicopathological entities of marginal zone lymphoma (MZL), including extranodal or mucosa-associated lymphoid tissue (MALT) lymphoma and MZL of nodal (NMZL) or splenic (SMZL) type. Of these, leukemic presentation, usually as small or villous lymphocytes, is more common in SMZL, while leukemic change in NMZL is rare, and the morphology has not been characterized. We present a stage 4 MZL involving lymph node, spleen, and bone marrow with two relapses after chemotherapy. The leukemic cells at the second relapse revealed irregular nuclear contours with multilobated nuclei (so-called flower cells or floral cells) mimicking the neoplastic cells in adult T-cell leukemia/lymphoma (ATLL). The absence of leukemic change and splenic hilar lymphadenopathy at initial presentation, expression of IgD by tumor cells, and cytogenetic changes of +7 suggested that this tumor might be a NMZL. Although the cytomorphologic features of floral leukemic cells might suggest ATLL, thorough clinical and laboratory workup helped to reach a correct diagnosis. Our findings broaden the cytological spectra of leukemic cells in MZL and illustrate the importance of immunophenotyping. PMID- 17913386 TI - Primary pleural thymoma with coexistent incidental small hepatocellular carcinoma - an autopsy case report with brief review of literature. AB - Primary pleural thymomas are rare tumors often mistaken for malignant mesothelioma clinically and radiologically. An autopsy case report of primary pleural thymoma associated with a coincidental small hepatocellular carcinoma is presented. This case is reported because of the rarity of pleural thymoma and the coincidental finding of a small hepatocellular carcinoma in a non-cirrhotic background. The literature on these two tumors has been reviewed. PMID- 17913387 TI - Chemical constituents of Roldana lineolata. AB - The new eremophilanolide 1, the known eremophilane derivatives 2 and 3, already described as part of mixtures, the known compounds hyperin, 2''-acetylhyperin and two calenduladiol esters were isolated from Roldana lineolata. Their structures were determined by spectroscopic methods. Compounds 1 and 2 as well as their acetylated and oxidized derivatives were tested against several fungi strains. Eremophilanolide 1 showed a mild activity against Trichophyton mentagrophytes. PMID- 17913388 TI - [Molecular fingerprint of bacterial communities and 16S rDNA intra-species heterogeneity: a pitfall that should be considered]. AB - Molecular fingerprinting methods are currently used to study microbial communities by culture independent approaches. They are proposed as identification tool owing to the availability of rapid automated methods. The 16S rRNA gene (16S rDNA) is an efficient marker for bacterial identification and microbial communities analysis. However, the 16S rDNA polymorphism among strains of the same species is an underestimated pitfall of the fingerprinting approaches. AIM OF THE STUDY: We studied the 16S rDNA variability among strains of three bacterial species of medical interest. MATERIAL AND METHODS: Total DNA was extracted from clinical isolates of Pseudomonas aeruginosa (N=20), Clostridium difficile (N=20) and Enterobacter cloacae (N=14). The Polymerase Chain Reaction (PCR) products obtained with consensus primers flanking the 16S rDNA variable regions V3 and V6-V7-V8 were separated by Temporal Temperature Gradient gel Electrophoresis (TTGE). DNA extracted from TTGE bands were sequenced and analysed. RESULTS: All the isolates of P. aeruginosa and of C. difficile displayed one single TTGE band with constant migration distances suggesting that there was no 16S rDNA polymorphism among strains in these two species. Oppositely, the isolates of E. cloacae gave complex TTGE patterns formed by multiple bands with variable migration distances. These patterns corresponded to 16S rRNA genes variable in a single genome as well as among strains of the species. CONCLUSION: Intra-species and/or intragenomic variability of 16S rDNA should be taken into account for pertinent interpretation of molecular fingerprint. For this purpose, a comprehensive description of the polymorphism of this marker is necessary. PMID- 17913389 TI - [Serotonin modulates HIV replication in primary culture of human macrophages: involvement of 5-HT(1A) sub-type receptors]. AB - The neurotransmitter 5-hydroxytryptamine (5-HT), commonly known as serotonin, is released at peripheral sites from activated platelets. At inflammatory sites, macrophages and lymphocytes could be exposed to 5-HT concentrations up to 100 microM. Moreover, 5-HT could modulate cytokine secretion by monocytes/macrophages and immune functions through the uptake of 5-HT at these inflammatory sites from T cells and dendritic cells. HIV infection is also under the control of inflammatory processes (including T cell proliferation and cytokines secretion). On this basis, we studied explored herein the effects of 5-HT on HIV-1/Ba-L (macrophage-tropic virus) replication in primary cultures of human macrophages. This pharmacological study with isotype-selective receptor agonists and antagonist allowed us to show that the 100 microM 5-HT concentration via 5-HT(1A) subtype receptors could decrease HIV replication. This observation was associated with an increase of MIP-1alpha secretion such as an increase of MIP-1alpha mRNA production and with a decrease of HIV-coreceptor CCR5 cell surface expression. Our results point out for the first time the inhibitory effects of 5-HT on HIV replication in primary culture of human macrophages via activation of 5-HT(1A) subtype receptors. PMID- 17913390 TI - [Chagas disease: screening tests evaluation in a blood military center, prevalence in the French Army]. AB - Chagas disease is a major public health problem in Latin and Central America, 15 to 20 million people are affected and some 100 million is at risk of acquiring Chagas disease. Chagas disease starts to appear in amazonian area and french Guyana. Three kits: Elisa Novagnost (Dade Behring), BioElisa Chagas (Orgentec) et Elisa Cruzi (BioMerieux) were compared using performance panel. Sensibility, reproductibility and specificity (using 40 serum of blood donors who never went to an endemic area) were evaluated. Orgentec assay (recombinant antigens) and BioMerieux assay (whole-epimastigote antigens) performed better than Dade Behring assay. The latter was discarded from the study at this stage. Lack of sensibility seems due to the antigenic composition. Reproductibility and specificity are good for the other two tests. Mixtures of recombinants antigens increased specificity, but sensibility is better using mixtures of whole-epimastigote antigens. For routine blood donor screening both tests must be performed. A prevalence study was done during 11 months on 1570 serum of military blood donors. Despite of a low prevalence (less than 0.7 per thousand), the entire donation from donors who were in the endemic area (7.95% from our whole population) are screened for antibody against Trypanosoma cruzi, with these two assays. PMID- 17913391 TI - [Evaluation of IDI-MRDA assay on a collection of community-acquired methicillin resistant Staphylococcus aureus isolates and on carriage specimens]. AB - The efficacy of infection control measures against MRSA is linked to the rapid detection of MRSA. With the conventional diagnosis by culture the response delays vary from 48 to 72 hours. In contrast molecular techniques give results within hours. OBJECTIVE: The objective of the present study is to perform the IDI-MRSA PCR test (BD Diagnostic GeneOhm) on a collection of characterized community acquired MRSA (CA-MRSA) isolates and on carriage specimens. COLLECTION OF ISOLATES: Fifty-two isolates of CA-MRSA previously characterised by their toxinotype and SCCmec type cassette were analysed. All of them were identified as MRSA by the IDI-MRSA test. SPECIMENS: Seventy screening specimens from 35 different patients were tested in comparison with the culture on specific media (MRSA ID, BioMerieux). Among those 70 specimens, 24 were from nose, 25 from cutaneous sites (axillar; groin) and 21 from other sites. Sensitivity and specificity were 86.4 and 91.3% respectively; positive and negative predictive values were 93.3 and 82.6% respectively. RESULTS: Three of four false-positive results came from specimens collected during a decolonisation treatment. Without taking account those specimens, specificity and positive predictive reach 97.9 and 95% respectively. This study shows that IDI-MRSA is an interesting additional test for the diagnosis of MRSA carriage. PMID- 17913392 TI - [Microelectrophoresis and atomic force microscopy: new tools to explore mechanism of action of antibacterial compounds]. AB - Microbial cell surface properties play a central role in controlling phenomena such as bacterial adhesion and biofilm formation (on stent or on prosthesis for example). The quantification of these properties and the understanding of interactions with antibacterial compounds remain difficult, in view of the complex and dynamic nature of the cell wall constituents. Various approaches, macroscopic, microscopic or molecular, have been developed. Two of them interest us today: (i) microelectrophoresis, which permits to evaluate surface modifications by measuring eletrophoretic mobility; and (ii) atomic force microscopy (AFM), a high resolution imaging device, which allows investigations at nanometric scale. After brief presentation of principles and instrumentations, the aim of this article is to present the different applications of these techniques in Microbiology, and to discuss interest of these tools in order to investigate mechanism of action of antibacterial compounds. PMID- 17913393 TI - Mediterranean-spotted fever: clinical and laboratory characteristics of 34 children in Oran (Algeria). AB - INTRODUCTION: Mediterranean-spotted fever (MSF) is endemic in the Mediterranean basin. The aim of our study is to describe clinical and laboratory characteristics of MSF in Algeria. METHODS: Retrospective study of 34 children suffering from MSF hospitalized from 2002 to 2005. Diagnosis and outcome data were abstracted from patient medical reports. The serological confirmation was obtained by immunofluorescent antibody. RESULTS: All the children were hospitalized during summer time. The median age was 4.5 years. The sex ratio in the children was 0.76. Clinical features were high fever, maculopapulous rash and black eschar. Among analytical findings, leucocyte count was decreased in 50% of children during the first week of illness. The indirect immunofluorescence for Rickettsia conorii was positive in 70% of cases in the second determination. All received antimicrobial therapy. All children cured without complications. CONCLUSION: MSF is a mild rickettsial disease in Algeria. PMID- 17913394 TI - Rapid identification of bacteria, mecA and van genes from blood cultures. AB - The Genotype technology, a quick molecular genetic assay based on DNA multiplex amplification with biotinylated primers followed by hybridization to membrane bound probes, complies with the requirements for a fast diagnosis of sepsis. We evaluated the new Genotype BC Gram-negative and Gram-positive test kits (Hain Life Science, Germany) which respectively allow for the identification of 15 species of Gram-negative (GN) rods, and the identification of 17 Gram-positive (GP) bacteria species together with the determination of methicillin and vancomycin resistance (mecA and van genes). The study was performed on 60 positive blood cultures from BacT/ALERT bottles (aerobic, anaerobic and pediatric bottles). First, a Gram stain was carried out to select between Genotype BC GP or GN test, then identification were performed by the Genotype BC tests and by biochemical conventional tests after subculture and phenotypic susceptibility determination. The operating procedure was very easy to carry out and required a small amount of starting material (5 to 10 microL of blood culture). The results were available within 4.5 hours. For all the blood cultures, the Genotype BC results correlated with the biochemical identification and phenotypic antibiotics susceptibility. According to our results, this DNA strip technology based assay can easily be incorporated into routine diagnosis. PMID- 17913395 TI - Prescription opioid use, misuse, and diversion among street drug users in New York City. AB - OBJECTIVE: The use of heroin, cocaine, and other drugs is well researched in New York City, but prescription opioids (POs) have been overlooked. This study documents patterns of PO use, misuse, and diversion among street drug users, and begins to indicate how drug culture practices interact with the legitimate therapeutic goals of PO prescriptions (e.g. pain management). METHODS: Staff completed interviews inquiring about the reasons for use of POs and illicit drugs with 586 street drug users. Ethnographers wrote extensive field notes about subjects' complex patterns of PO use. RESULTS: Methadone was used (71.9%) and sold (64.7%) at a higher level than OxyContin, Vicodin, and Percocet, used by between 34% and 38% of the users and sold by between 28% and 41% of the sellers. Recent PO use is associated with the recency of using heroin and cocaine (p<.001). Half of the heroin/cocaine sellers sold POs, and one quarter of the PO sellers only sold POs. Subjects were classified into four groups by whether they diverted POs or used POs to relieve pain or withdrawal rather than for euphoria. This classification was associated with frequency of PO use, whether POs were obtained from doctors/pharmacies or from drug dealers and family members, and those mostly likely to use POs for pain and withdrawal. CONCLUSIONS: POs are an important component of street drug users' drug-taking regimes, especially those who are Physically Ill Chemical Abusers (PICA). Future research is needed to model PO use, misuse, and diversion among this population. PMID- 17913396 TI - The association of self-reported neighborhood disorganization and social capital with adolescent alcohol and drug use, dependence, and access to treatment. AB - AIMS: This research examines adolescent perceptions of neighborhood disorganization and social capital to determine if they are associated with adolescent alcohol or drug (AOD) use, AOD dependence, and access to AOD treatment. DESIGN: This is a secondary analysis of data from the 1999 and 2000 National Survey on Drug Use and Health (NSDUH). The NSDUH is a cross-sectional survey of a random sample of the non-institutionalized United States population and is conducted in respondents' homes. PARTICIPANTS: Youth between the ages of 12 and 17, yielding a sample size of 38,115 respondents. MEASUREMENTS: Neighborhood disorganization was self-reported by youth in response to eight items; 10 items measured social capital. AOD use was also self-reported. AOD dependence was assessed by a series of questions regarding symptoms and impairment that is consistent with the criteria specified in the DSM-IV. RESULTS: A little more than half of the youth reported never using alcohol or drugs (54.3%), 41.1% reported lifetime AOD use, and 4.6% were AOD dependent. Two percent reported receiving AOD treatment. Medium and high levels of social capital were negatively associated with AOD use and dependence. Social capital was unrelated to access to AOD treatment. Neighborhood disorganization was positively associated with AOD use, dependence, and access to treatment. CONCLUSIONS: After controlling for individual- and family-level characteristics, neighborhood disorganization and social capital were associated with AOD use and dependence. The findings suggest that subjective measures of social context may be an important component of the complex biopsychosocial model of adolescent AOD addiction and treatment utilization. PMID- 17913397 TI - Identification of a novel Bcl-2 promoter region that counteracts in a p53 dependent manner the inhibitory P2 region. AB - Expression of the anti-apoptotic proto-oncogene bcl-2 is negatively affected by the pro-apoptotic p53. To understand the regulation of bcl-2 expression by p53, we studied the bcl-2 promoter regions individually and in the context of the full length promoter. While the P1 promoter displayed the highest p53-independent activity, the P2 promoter activity was suppressed in p53-sufficient cancer cell lines. In addition, P2 activity was higher in primary airway epithelial cells from p53(-/-) mice compared to those from p53(+/+) mice. Chromatin immunoprecipitation assays confirmed that p53 interacts within a 140 bp sequence of P2 that contained the CCAAT- and TATA-elements. However, when P1 and P2 are linked in one construct, P2 suppressed P1 activity independent of p53. A potential novel promoter with a p53-dependent activity was identified located between P1 and P2, and was designated M. In the context of the full-length bcl-2 promoter, M counteracted in a p53-dependent manner the suppressive activity of P2 on P1. Collectively, these data suggest that P1 promoter is the main driving force for transcribing the bcl-2 gene and P1 activity is modulated by M and P2 in a p53-dependent and -independent manner. These findings may have implications for therapies that are geared towards inhibiting bcl-2 gene expression and inducing cell death. PMID- 17913398 TI - Cyclic equibiaxial tensile strain induces both anabolic and catabolic responses in articular chondrocytes. AB - Mechanical disturbance is directly implicated in the development of osteoarthritis (OA) but the precise mode for degenerative changes is still largely unknown because of the complexity of the biomechanical and biochemical milieu in the articular joint. To investigate the effects of tensile strain on articular cartilage, cyclic equibiaxial tensile strain (CTS, 0.5 Hz, 10% strain) was applied to monolayer cultures of porcine articular chondrocytes by using a Flexercell strain unit. Overproduction of proinflammatory mediators and imbalanced expression of anabolic and catabolic genes were induced. The cellular secretion of nitric oxide (NO) and prostaglandin E(2) (PGE(2)), as well as the mRNA level of cyclooxygenase-2 (COX-2) were up-regulated in response to mechanical stimuli. Additionally, CTS resulted in an initial peak of anabolic response at 3 h of stretch with respect to the expression of type II collagen and aggrecan. After 12 h of CTS, the expression for these two cartilage-specific matrix proteins fell to control levels. A distinct catabolic response developed after 24 h of stretch with an increase in matrix metalloproteinase-1 (MMP-1). Interestingly, a parallel increase in transforming growth factor (TGF) beta3 was associated with the anabolic changes while an increase in expression of TGF beta1, the predominant isoform of the TGF family, appeared at 24 h. The expression at 24 h of MMP-1, an enzyme that degrades interstitial collagens as well as other cartilage matrix proteins and TGF beta1, may signify a shift towards matrix remodeling and potentially a change in matrix composition as a consequence of continuous CTS. PMID- 17913399 TI - Fusobacterium equinum possesses a leukotoxin gene and exhibits leukotoxin activity. AB - Fusobacterium equinum, a gram negative, rod-shaped and an obligate anaerobic bacterium is a newly described species. The organism is associated with necrotic infections of the respiratory tract in horses that include necrotizing pneumonia, pleuritis and paraoral infections. The species is closely related to F. necrophorum that causes liver abscesses in cattle and sheep, calf-diphtheria in cattle, and foot-rot in sheep and cattle. Leukotoxin, an exotoxin, is an important virulence factor in bovine strains of F. necrophorum. Our objective was to examine strains (n=10) of F. equinum for leukotoxin (lktA) gene and its toxic effects on equine leukocytes. Southern hybridization and partial DNA sequencing revealed that all the 10 strains had the lktA gene with greater similarities to F. necrophorum subsp. necrophorum. The secreted leukotoxin was detected in the culture supernatant and its biological activity was determined by viability assays with equine polymorphonuclear cells (PMNs) using flow cytometry. While culture supernatants of four strains (E1, E7, E9, and E10) were highly toxic to equine PMNs; strain E5 was moderately toxic and the remaining strains (E2, E3, E4, E6, and E8) were only mildly toxic. Our data indicated that F. equinum isolates had lktA gene and its product was toxic to equine leukocytes. Therefore, leukotoxin may be an important virulence factor in F. equinum infections. PMID- 17913400 TI - Detection and differentiation of re-emerging fowlpox virus (FWPV) strains carrying integrated reticuloendotheliosis virus (FWPV-REV) by real-time PCR. AB - Current strains of fowlpox virus (FWPV) carrying circulating reticuloendotheliosis virus (FWPV-REV) sequence are becoming more pathogenic to poultry. This is evidenced by the fact that vaccination with current available FWPV vaccines provides limited protection against them. To characterize REV insertions in a collection of both older and more recent field isolates, we developed three different types of adjacent oligoprobes and primer sets from specific genomic locations of FWPV and REV: REV-ENV (accession no. K02537, 1382 2260), FWPV-REV integration site (accession no. AF006064, 86-1328), FWPV (accession no. AF198100, 232461-232670), and REV-LTR (accession no. V01204, 305 496). The data indicated that the primers from the REV-ENV region and the TaqMan probes specifically targeted REV-ENV sequences of FWPV-REV strains. Furthermore, the strains were differentiated based on quantitative melting temperature (T(m)) of their amplified products using FRET-based probes. The amplified products were further characterized by sequencing and multiple sequence alignment analysis. The results suggest that integrated REV-ENV sequences are both common and mostly conserved in field isolates. However, the minor variations found within the short targeted ENV sequence from FWPV-REV strains suggest that these strains could have either undergone periodic point mutational changes or integration with different REV-ENV subtypes. PMID- 17913401 TI - Mycobacterium pinnipedii: transmission from South American sea lion (Otaria byronia) to Bactrian camel (Camelus bactrianus bactrianus) and Malayan tapirs (Tapirus indicus). AB - Tuberculosis infections caused by Mycobacterium (M.) pinnipedii in a South American sea lion, Bactrian camel, and Malayan tapirs kept in two zoological gardens spanning a time period of 5 years are reported. The zoos were linked by the transfer of one tapir. Conventional bacteriological and molecular methods were applied to detect the pathogen. Spoligotyping and MIRU/VNTR-typing performed to assess the genetic similarity revealed identical molecular characteristics of the isolates from all animals involved. Anti-tuberculosis antibodies were detected using ELISA and a recently developed serological rapid test. The study shows that: (i) using molecular methods, the assessment of the genetic relationship of infectious agents helps to confirm the routes of infection, and that (ii) immunological tests may help to detect tuberculosis infections ante mortem more reliably and early. This would prevent the transfer of tuberculosis by asymptomatic animals. PMID- 17913402 TI - Best practice guidelines for the operation of a donor human milk bank in an Australian NICU. AB - Until the establishment of the PREM Bank (Perron Rotary Express Milk Bank) donor human milk banking had not occurred in Australia for the past 20 years. In re establishing donor human milk banking in Australia, the focus of the PREM Bank has been to develop a formal and consistent approach to safety and quality in processing during the operation of the human milk bank. There is currently no existing legislation in Australia that specifically regulates the operation of donor human milk banks. For this reason the PREM Bank has utilised existing and internationally recognised management practices for managing hazards during food production. These tools (specifically HACCP) have been used to guide the development of Standard Operating Procedures and Good Manufacturing Practice for the screening of donors and processing of donor human milk. Donor screening procedures are consistent with those recommended by other human milk banks operating internationally, and also consistent with the requirements for blood and tissue donation in Australia. Controlled documentation and record keep requirements have also been developed that allow complete traceability from individual donation to individual feed dispensed to recipient and maintain a record of all processing and storage conditions. These operational requirements have been developed to reduce any risk associated with feeding pasteurised donor human milk to hospitalised preterm or ill infants to acceptable levels. PMID- 17913403 TI - Both postnatal and postmenstrual age contribute to the interindividual variability in tramadol glucuronidation in neonates. AB - INTRODUCTION: Although of pharmacokinetic and -dynamic relevance, data on ontogeny of UDP-glucuronosyltransferase (UGT) activity in neonates are scant. We therefore wanted to assess the impact of both postnatal and postmenstrual age (PNA/PMA) on the interindividual variability of glucuronidation to overall tramadol urinary elimination in neonates. METHODS: O-demethyl tramadol (M1) and M1-glucuronide (M1G) were determined in 24 hour urine collections during continuous intravenous tramadol administration in neonates. Glucuronidation fraction (%) was calculated by the ratio of M1G to the sum of M1G and M1 free (M1total). Fractions (%) in early (0.05). After 10 months, there was a significant reduction in frequency of hot flushes among isoflavone users when compared to those on placebo (3.1 +/- 2.3 and 5.9 +/- 4.3, respectively) (p<0.001). Kupperman index mean values showed a significant reduction in both groups. However, soy isoflavone was significantly superior to placebo, in reducing hot flush severity (69.9% and 33.7%, respectively) (p<0.001). Endometrial thickness, mammography, vaginal cytology, lipids and hormonal profile did not change in both groups. No serious adverse event related to isoflavone treatment was reported. CONCLUSIONS: The soy isoflavone extract exerted favorable effects on vasomotor symptoms and good compliance, providing a safe and effective alternative therapeutic for postmenopausal women. PMID- 17913409 TI - Effect of surface charge and density of distearylphosphatidylethanolamine-mPEG 2000 (DSPE-mPEG-2000) on the cytotoxicity of liposome-entrapped ricin: effect of lysosomotropic agents. AB - Ricin was encapsulated in various liposomes having neutral, negatively and positively charged and different density of DSPE-mPEG-2000 on the surface and cytotoxicity of ricin entrapped in these different charged liposomal formulations was studied in CHO pro(-) cells and compared with free ricin with a view to develop an optimum delivery system for ricin in vivo. It was observed that the cytotoxicity of ricin entrapped in various charged liposomes was significantly dependent on the charge on the surface of liposomes. The maximum cytotoxicity of ricin was observed when it was delivered through negatively charged liposomes. Monensin enhances the cytotoxicity of ricin entrapped in various charged liposomes and the extent of enhancement of the cytotoxicity is significantly dependent on the charge on the surface of liposomes. Maximum potentiation (213.14 fold) of cytotoxicity of ricin was observed when it was delivered through positively charged liposomes followed by negatively charged (83.36-fold) and neutral (71.30-fold) liposomes, respectively. Studies on the kinetics of inhibition of protein synthesis by ricin entrapped in various charged liposomes revealed that lag period of inhibition of protein synthesis is significantly lengthened following delivery through various charged liposomes. However, in the presence of monensin, the lag period was reduced. There is a marginal variation in the cytotoxicity of ricin entrapped in various charged liposomes after incorporation of 5mol% of DSPE-mPEG-2000 on the surface. However, there is a significant variation in the enhancing potency of monensin on the cytotoxicity of ricin entrapped in various charged liposomes in CHO pro(-) cells following incorporation of 5mol% DSPE-mPEG-2000 on the surface. Studies on the effect of variation of density of DSPE-mPEG-2000 on the surface of various charged liposomes on the enhancement of cytotoxicity of entrapped ricin by monensin in CHO pro(-) cells showed that the enhancing potency of monensin on the cytotoxicity of ricin entrapped in various charged liposomes is significantly dependent on the density of DSPE-mPEG-2000 on their surface. It was also observed that the efficacies of monensin on the enhancement of cytotoxicity of ricin entrapped in various charged PEG-liposomes in CHO pro(-) cells was highly related to their amount of cell-association. The present study has clearly shown that by suitable alteration of liposomal lipid composition, charge and density of hydrophilicity it would be possible to direct liposomal ricin to specific cells for their selective elimination in combination with monensin. PMID- 17913412 TI - Sulfonamides as a subject to study molecular interactions in crystals and solutions: sublimation, solubility, solvation, distribution and crystal structure. AB - Crystal structures of 4-amino-N-(4-chlorophenyl)-benzene-sulfonamide (IV), 4 amino-N-(2,3-dichlorophenyl)-benzene-sulfonamide (V), 4-amino-N-(3,4 dichlorophenyl)-benzene-sulfonamide (VI) and 4-amino-N-(2,5-dichlorophenyl) benzene-sulfonamide (VII) were solved by X-ray diffraction method. Temperature dependencies of saturated vapour pressure and thermodynamic functions of sublimation process were calculated (IV: delta Gsub298=74.0 kJ mol(-1), delta Hsub298=134.1+/-1.2 kJ mol(-1), delta Ssub298=202+/-3 J mol(-1)K(-1); V: delta Gsub298=61.7 kJ mol(-1), delta Hsub298=141.1+/-0.7 kJ mol(-1), delta Ssub298=266+/-2 J mol(-1)K(-1); VI: delta Gsub298=85.8 kJ mol(-1), delta Hsub298=167.5+/-3.6 kJ mol(-1), delta Ssub298=274+/-8 J mol(-1)K(-1); VII: delta Gsub298=75.7 kJ mol(-1), delta Hsub298=155.4+/-1.6 kJ mol(-1), delta Ssub298=268+/-4 J mol(-1)K(-1)). Thermochemical parameters of fusion and evaporation processes for the compounds were obtained. Temperature dependencies of the solubility in water, n-octanol were measured. The thermodynamic functions of solubility and solvation processes were deduced. The transfer processes of the molecules from water to n-octanol were analysed by diagram method and main driven forces were established. PMID- 17913411 TI - PLGA nanoparticles simultaneously loaded with vincristine sulfate and verapamil hydrochloride: systematic study of particle size and drug entrapment efficiency. AB - PLGA nanoparticles simultaneously loaded with vincristine sulfate (VCR) and verapamil hydrochloride (VRP) were prepared via combining O/W emulsion solvent evaporation and salting-out method. Ten independent processing parameters and two materials characteristics were assessed systematically to enhance the incorporation of the two hydrophilic low molecular weight drugs into PLGA nanoparticles and minimize nanoparticles size. Approaches investigated for the enhancement of drug entrapment efficiencies and the minimization of particle size included the influence of the molecular weight (MW) of PLGA and the lactide to glycolide (L:G) ratio of PLGA, PLGA concentration, the degrees of hydrolyzation and polymerization of PVA, PVA concentration, initial VCR and VRP content, acetone to dichloromethane volume ratio, aqueous phase pH, salt concentration of aqueous phase, aqueous to organic phase volume ratio, sonication time, sonication energy and removal rate of organic solvents. The nanoparticles produced by optimal formulation were submicron size (111.4+/-2.35nm, n=3) and of low polydispersity (0.062+/-0.023, n=3). Nanoparticles observed by transmission electron microscopy (TEM) showed extremely spherical shape. The entrapment efficiencies determined with high performance liquid chromatogram (HPLC) by ultracentrifuge method were 55.35+/-4.22% for VCR and 69.47+/-5.34% for VRP, respectively (n=3). PMID- 17913410 TI - Permeability of surface-modified polyamidoamine (PAMAM) dendrimers across Caco-2 cell monolayers. AB - Aim of this study was to prepare polyamine-conjugated PAMAM dendrimers and study their permeability across Caco-2 cell monolayers. Polyamines, namely, arginine and ornithine were conjugated to the amine terminals of the G4 PAMAM dendrimers by Fmoc synthesis. The apical-to-basolateral (AB) and basolateral-to-apical (BA) apparent permeability coefficients (P(app)) for the PAMAM dendrimers increased by conjugating the dendrimers with both of the polyamines. The enhancement in permeability was dependent on the dendrimer concentration and duration of incubation. The correlation between monolayer permeability and the decrease in transepithelial electrical resistance (TEER) with both the PAMAM dendrimers and the polyamine-conjugated dendrimers suggests that paracellular transport is one of the mechanisms of transport across the epithelial cells. Cytotoxicity of the polyamine-conjugated dendrimers was evaluated in Caco-2 cells by MTT (methylthiazoletetrazolium) assay. Arginine-conjugated dendrimers were slightly more toxic than PAMAM dendrimer as well as ornithine-conjugated dendrimers. Though investigations on the possible involvement of other transport mechanisms are in progress, results of the present study suggest the potential of dendrimer polyamine conjugates as drug carriers to increase the oral absorption of drugs. PMID- 17913413 TI - Assessment of the phytochemical and antitrypanosomal properties of some extracts of leaves, stem and root bark of Landolphia sp., P. Beauv. AB - INTRODUCTION/JUSTIFICATION: There is urgent need to source for alternative chemotherapy against trypanosmosis, a disease of major importance in human and economic animals. MATERIALS AND METHODS: Therefore, petroleum ether, chloroform, methanol and aqueous extracts sequentially obtained from the leaves, stem and root barks of Landolphia uniflora were evaluated for their in vitro and in vivo antitrypanosomal activities against Trypanosoma brucei brucei, as well as their phytochemical constituents. RESULTS: Steroids and triterpenes, resins, tannins, saponins and flavonosides were detected in almost all the extracts, but alkaloid was absent in methanol extract of the stem and stem, as well as the chloroform extract of the root bark. In vitro, all extracts of the roots displayed significant antitrypanosomal activity, while only the chloroform extracts of the leaves and stem bark showed activity at both test concentrations (4 and 2 mg/ml). However, under in vivo condition, the methanol extracts showed the greatest activity, eliminating parasitaemia within the 10 days treatment period and prolonging survival period at 200 and 300 mg/kg body weight intramuscular doses. CONCLUSION: These results suggest that Landolphia uniflora could be useful in the management of trypanosomiasis. PMID- 17913414 TI - Inhibition of vascular smooth muscle cell proliferation by serum from rats treated orally with Gastrodia and Uncaria decoction, a traditional Chinese formulation. AB - AIM: This study was to investigate the effect of serum from rats treated orally with GUD on vascular smooth muscle cells (VSMCs) proliferation in vitro. METHODS: Cell proliferation was measured by Methyl thiazolyl tetrazolium (MTT) assay. Expression of proliferating cell nuclear antigen (PCNA) and proto-oncogene c-myc were measured by immunochemical staining and image analysis. Griess reagent were used to detect nitric oxide (NO) level. Endothelin-1 (ET-1) level was measured by radioimmunoassay. RESULTS: GUD serum (2.5%-10%) inhibited VSMCs proliferation in a dose and time-dependent manner. GUD serum inhibited the expression of PCNA and c-myc. Moreover, GUD serum increased nitric oxide (NO), and decreased Endothelin 1 (ET-1) level in culture medium. CONCLUSION: GUD serum exhibited directly inhibitory effect in VSMCs proliferation. Inhibiting the expression of PCNA and c myc, increasing NO level and decreasing ET-1 level might be associated with the antiproliferative effect. PMID- 17913415 TI - Spasmolytic action of Lepechinia caulescens is through calcium channel blockade and NO release. AB - The present study was undertaken to elucidate the mode of action of methanol extract from aerial parts of L. caulescens (TC-MELc) as spasmolytic agent on in vitro rat ileum test, and investigate the possible antibacterial activity of different extracts from the plant. TC-MELc induced a concentration-dependent (0.001 to 100microg/mL) antispasmodic effect on spontaneous contractions. TC-MELc also (IC50 11.2microg/mL) induced a marked depression on cumulative concentration response curve for carbachol (Emax=2.3+/-0.3g vs. 0.66+/-0.1g) and serotonin (Emax=1.1+/-0.3g vs. -0.01+/-0.09g). Besides, extract decreased and displaced to the right KCl and CaCl2 concentration-response curves. Moreover, TC-MELc (11.2microg/mL) provoked a total relaxation when ileum strips were contracted with carbachol (1microM) in calcium-free Krebs solution. Pre-treatment with l NAME (10microM) produced a significant change of the relaxant response and activity was markedly inhibited. Additionally, hexanic (HELc), dichloromethanic (DELc) and methanolic (MELc) extracts from aerial parts were studied to determine their antibacterial activity. DELc showed antibacterial activity on all bacterial strains assayed (40% height loss) vertebral fractures between T1 and L1 was determined using an established semi-quantitative method and confirmed by morphological measurement. Results were compared with the formal CT report. RESULTS: Scans of 192 patients were analysed (95 female; 97 male); mean age 70.1 years. Thirty eight (19.8%) patients had one or more moderate to severe vertebral fractures. Only 5 (13%) were correctly reported as having osteoporotic fractures in the official report. The sensitivity of axial CT images to vertebral fracture was 0.35. CONCLUSION: Incidental osteoporotic vertebral fractures are under-reported on CT. The sensitivity of axial images in detecting these fractures is poor. Sagittal reformations are strongly recommended to improve the detection rate. PMID- 17913430 TI - Chromatographic analysis and decomposition product characterization of compound SR16157 (NSC 732011). AB - SR16157 (21-(2-N,N-diethylaminoethyl)oxy-7alpha-methyl-19-norpregna-1,3,5(10) triene-3-O-sulfamate) is a novel, dual-acting estrone sulfatase inhibitor currently in preclinical development for use in breast cancer therapy. The compound has a dual mechanism of action: the sulfamate-containing parent compound SR16157 inhibits estrogen biosynthesis by irreversibly inhibiting the enzyme estrone sulfatase. The phenolic metabolite, SR16137, generated by the sulfatase enzyme is a potent antiestrogen in breast tissues and has beneficial effects in bone and the cardiovascular system. As part of the ongoing preclinical studies, an HPLC assay method has been developed and validated for SR16157. The assay method is specific, accurate (recovery=99.4-101.1), linear (r(2)> or =0.9999), precise (intraday R.S.D.< or =1.1%, intermediate R.S.D.< or =0.8%), and sensitive (limit of detection=1.0 microg/ml). It separates SR16157 from its impurities and forced decomposition products, which have been characterized by LC coupled with mass and UV spectral data. Major decomposition pathways are hydrolysis, hydroxylation, and oxidation. PMID- 17913431 TI - Detection limit of measurement of pharmaceuticals labeled with short-lived isotopes in HPLC with flow-through gamma-counter. AB - This paper proposes a method for estimating the detection limit, which is defined as 3.3 times the standard deviation (S.D.) of blank measurements under the situations where the repetition of measurement is difficult or impossible because of a short half-life of radioactivity. The FUMI theory, which can estimate an S.D. value without repetition in various instrumental analyses, is adopted and proved here to be available in a radio-HPLC system as well. (99m)Tc-ECD (T(1/2)=360.6 min) that is a lipophilic compound for the diagnosis of regional brain perfusion is taken as an example. PMID- 17913432 TI - Fast gas chromatography and negative-ion chemical ionization tandem mass spectrometry for forensic analysis of cannabinoids in whole blood. AB - The present work describes a fast gas chromatography/negative-ion chemical ionization tandem mass spectrometric assay (Fast GC/NICI-MS/MS) for analysis of tetrahydrocannabinol (THC), 11-hydroxy-tetrahydrocannabinol (THC-OH) and 11-nor-9 carboxy-tetrahydrocannabinol (THC-COOH) in whole blood. The cannabinoids were extracted from 500 microL of whole blood by a simple liquid-liquid extraction (LLE) and then derivatized by using trifluoroacetic anhydride (TFAA) and hexafluoro-2-propanol (HFIP) as fluorinated agents. Mass spectrometric detection of the analytes was performed in the selected reaction-monitoring mode on a triple quadrupole instrument after negative-ion chemical ionization. The assay was found to be linear in the concentration range of 0.5-20 ng/mL for THC and THC OH, and of 2.5-100 ng/mL for THC-COOH. Repeatability and intermediate precision were found less than 12% for all concentrations tested. Under standard chromatographic conditions, the run cycle time would have been 15 min. By using fast conditions of separation, the assay analysis time has been reduced to 5 min, without compromising the chromatographic resolution. Finally, a simple approach for estimating the uncertainty measurement is presented. PMID- 17913433 TI - Detecting Blastocystis using parasitologic and DNA-based methods: a comparative study. AB - Few studies have targeted the relative performance of diagnostic methods used for the detection of Blastocystis, a unicellular organism often present in fecal specimens from individuals with and without gastrointestinal symptoms. Aims of this study included a comparison of the formol ethyl acetate concentration technique (FECT), permanent trichrome staining of feces fixed in sodium acetate acetic acid-formalin (SAF-PST), xenic in vitro culture (XIVC), and polymerase chain reaction (PCR) regarding Blastocystis screening of 107 samples from 93 patients with suspected enteroparasitic disease. Compared with PCR, the sensitivity/specificity of XIVC, SAF-PST, and FECT was 89%/100%, 82%/100%, and 50%/100%, respectively. False-negative results generated by the FECT and SAF-PST appeared to be associated with Blastocystis sp. subtype 3. A comparison of results obtained by dideoxy sequencing of positive PCR products amplified from DNA extracted directly from feces and DNA extracted from 5- and 28-day-old XIVC of 10 randomly chosen Blastocystis isolates showed no disparities, indicating that XIVC has very little or no impact on subtype distribution or variation within a given specimen. It is recommended that short-term XIVC be used for cost effective screening of fresh fecal specimens for Blastocystis infection to generate valid prevalence estimates and to identify isolates for molecular characterization in studies aiming to illuminate the molecular epidemiology of Blastocystis. PMID- 17913435 TI - Prevalence of clinical isolates of Escherichia coli and Klebsiella spp. producing multiple extended-spectrum beta-lactamases. AB - Eleven thousand two hundred seventy-two Escherichia coli, 1109 Klebsiella pneumoniae, 1124 Salmonella enterica, and 602 Klebsiella oxytoca unrelated clinical isolates were obtained between 2001 and 2004 in a university hospital in Salamanca, Spain. One hundred thirteen E. coli (1%), 32 K. pneumoniae (2.9%), 4 K. oxytoca (0.66%), and 5 S. enterica (0.44%) isolates produced extended-spectrum beta-lactamases (ESBLs). We obtained 42.2% of the ESBL-producing isolates from outpatients and 57.8% from inpatients. The most commonly detected ESBLs were CTX M 14 (43.5% of ESBL-producing isolates), TEM-116 (22.1%), and SHV-2 (15.6%). A CTX-M 27-producing E. coli is 1st reported in Spain in this study. Two (20 isolates, 13%) or 3 (7 isolates, 4.5%) ESBLs were produced by 17.5% of ESBL producing isolates (27 isolates). The most frequent combinations were CTX-M 14 + TEM-116 (5.7%), SHV-12 + TEM-116 (2.6%), and SHV-2 + CTX-M 14 + TEM-116 (2.6%). Clonal diversity was high even between isolates producing the same combinations of 2 or 3 beta-lactamases. PMID- 17913434 TI - Prevalence of extended-spectrum beta-lactamases in Proteus mirabilis in a Taiwanese university hospital, 1999 to 2005: identification of a novel CTX-M enzyme (CTX-M-66). AB - A total of 1574 nonduplicate Proteus mirabilis isolates collected at a Taiwanese hospital during 1999 to 2005 were analyzed for production of extended-spectrum beta-lactamases (ESBLs). Forty-four ESBL-producing isolates including 22 CTX-M 14, 18 CTX-M-3, 2 CTX-M-24, and 2 CTX-M-66 producers were detected, and the proportion of ESBL producers increased from 0.7% in 1999 to approximately 6% after 2002. CTX-M-66 is a novel variant of CTX-M ESBLs that differs from CTX-M-3 by a Ser to Asn change at amino acid position 23. Coresistances to aminoglycosides and ciprofloxacin were very common in the CTX-M-3 producers. The presence of ArmA-type or RmtB-type 16S rRNA methylase that confers high-level aminoglycoside resistance was detected in 12 CTX-M-3 producers and 4 CTX-M-14 producers. Twenty-four clones including an endemic CTX-M-14-producing clone were observed among the 44 ESBL producers by pulsed-field gel electrophoresis, suggesting that both horizontal transfer and clonal spread contributed to the increased prevalence of bla(CTX-M) in P. mirabilis. PMID- 17913436 TI - Performance of MRSA ID chromogenic medium for detection of methicillin-resistant Staphylococcus aureus directly from blood cultures and clinical specimens. AB - MRSA ID was evaluated to see its performance in identifying methicillin-resistant Staphylococcus aureus (MRSA) directly from blood culture bottles (n = 837), wound swabs (n = 112), and abscesses (n = 18). Each positive blood culture and clinical specimen was directly inoculated on MRSA ID and the culture media routinely used. The sensitivity of MRSA ID was 97.8% after 24 h and 100% after 48 h for blood cultures, and 88.9% after 24 h and 100% after 48 h for wound samples. The specificity was 99.7% after 24 h and 99.6% after 48 h for blood cultures, and 100% after 24 and 48 h for wound samples. Four strains with green colonies indicating MRSA on MRSA ID were identified as methicillin-susceptible S. aureus (MSSA) by conventional methods. Three of these MSSA strains showed negative results with the mecA polymerase chain reaction, and 1 strain harbored the mecA gene. Using MRSA ID with primary culture media should decrease the time (18-24 h) to report a positive result compared with conventional methods. PMID- 17913437 TI - Endoplasmic reticulum stress during the embryonic development of the central nervous system in the mouse. AB - In the present study, we have found evidence for ER stress occurring during development of the central nervous system in the mouse. Several ER-resident stress-regulated chaperones, such as calreticulin, glucose regulated protein 78, glucose regulated protein 94, ER protein 57 and protein disulfide isomerase, were expressed at higher levels in embryonic brain and retina, compared with adult tissues. In contrast, calnexin, a chaperone that is not regulated by stress was equally abundant in embryonic and adult tissues. We also detected unfolded protein response during embryonic development. Both eukaryotic translation initiation factor 2 alpha and its phosphorylated form were more abundant in embryonic brain and retina than in adult tissues. Spliced X-box binding protein-1 mRNA was detected in embryonic brain and retina, while it was absent in adult counterparts. Partially glycosylated form of activating transcription factor 6 alpha, another ER stress indicator, was detected predominantly in embryonic brain. Finally, apoptotic pathway components, caspase-7 and -12, were more abundant in embryonic brain than in adult. The pattern of expression of chaperones together with activation of the unfolded protein response factors suggests the presence of ER stress during development of brain and retina. Furthermore, our data suggest that ER stress-like mechanism may induce apoptosis via activation of the caspases during embryonic development of the central nervous system. PMID- 17913438 TI - Physical activity behavior after total hip arthroplasty (THA): a prediction based on patient characteristics. AB - OBJECTIVE: To determine to what extent aspects of patient characteristics (age, gender, family status, education and comorbidity) are predictive for the level of physical activity of persons with a total hip arthroplasty (THA). METHODS: A cross-sectional study including 372 patients. Demographics, comorbidity and physical activity behavior were assessed by means of a questionnaire and from medical records. Linear regression analysis was used to determine to what extent patient characteristics are predictive of level of physical activity. Binary logistic regression modeling was used to determine the extent to which patient characteristics are predictive in meeting international guidelines on health enhancing physical activity. RESULTS: Age, education and family status significantly predict level of physical activity (R(2)=0.19). Only gender significantly predicts meeting international guidelines on health-enhancing physical activity (OR=2.06, 95% CI 1.20-3.54). CONCLUSION: Patients at risk can be identified by means of patient characteristics. Increasing age, lower education and living alone are associated with a physically inactive lifestyle. PRACTICE IMPLICATIONS: Health care workers involved in the treatment of THA patients should lay an emphasis on the beneficial aspects of physical activity. PMID- 17913439 TI - Beliefs about medicines and self-reported adherence among pharmacy clients. AB - OBJECTIVES: To analyse any association between general beliefs about medicines and self-reported adherence among pharmacy clients. Further, to examine general beliefs about medicines by background variables. METHODS: The data were collected by questionnaires including the general section of the Beliefs about Medicines Questionnaire (BMQ), the self-reporting Medication Adherence Report Scale (MARS) and the following background variables: gender, age, education, country of birth and medicine use. The General BMQ measures beliefs about medicines as something harmful (General-Harm), beneficial (General-Benefit) and beliefs about how doctors prescribe medicines (General-Overuse). RESULTS: Of the 324 participating pharmacy clients, 54% were considered non-adherent. An association was found between General-Harm and adherence. Adherent behaviour and higher level of education were associated respectively with more beneficial and less harmful beliefs about medicines. Those born in the Nordic countries regarded medicines as more beneficial. Current users of herbal medicines and non-users of medicines were more likely to believe that doctors overprescribed medicines. CONCLUSIONS: General-Harm was associated with adherence to medication among Swedish pharmacy clients. Country of birth, education and medicine use influenced beliefs about medicines. PRACTICE IMPLICATIONS: Increased awareness of the patient's beliefs about medicines is needed among healthcare providers. We should encourage patients to express their views about medicines in order to optimize and personalize the information process. This can stimulate concordance and adherence to medication. PMID- 17913440 TI - Randomized study of the effect of video education on heart failure healthcare utilization, symptoms, and self-care behaviors. AB - OBJECTIVE: Adherence to self-care behaviors improves heart failure (HF) morbidity and life quality. We examined short-term impact of video education (VE) in addition to standard education (SE) on HF healthcare utilization and self-care behavior adherence. METHODS: One hundred and twelve hospitalized patients were randomly assigned to SE (n=53) or SE plus VE (n=59). Differences between groups were analyzed in patients who underwent 3-month follow-up (39 SE and 37 VE patients). Mean age was 60+/-14 years; mean HF length was 57 months. RESULTS: Three-month healthcare utilization was similar between groups but VE patients needed less extra diuretic dosing (P<0.02), received more HF literature (P<0.03), and had less healthcare team telephone communication (P<0.04). VE patients had greater sign/symptom reduction (P<0.04); especially related to edema (P<0.01) and fatigue (P<0.01) and initiated more actions for edema (P<0.05) and dyspnea (with exercise or rest, both P<0.01). Overall, VE patients had a higher mean self-care behavior score (P<0.01), reflecting greater self-care adherence. CONCLUSION: Video education prompts self-care behavior adherence to control worsening signs/symptoms of volume overload. During 3-month follow-up, utilization of most healthcare resources was unchanged. PRACTICE IMPLICATIONS: VE is a useful adjunct to in-person education. PMID- 17913441 TI - Association of Harris lines and shorter stature with ethanol consumption during growth. AB - Ethanol consumption may impair bone growth. Transverse radiopaque lines (Harris lines) have been interpreted as manifestations of bone growth arrest due to nutritional stress. It is possible that ethanol consumption during growth leads to Harris lines formation and to a shorter stature. Plain X-ray film of the right tibia was performed to 175 individuals, who were inquired about ethanol consumption, periods of perceived hunger, and protracted illness during growth period (from birth to 18 years of age). Stature was also recorded. Men who drank during growth showed a shorter stature than those who did not (t=3.65, P<.001). Differences were not statistically significant among women (t=0.95). Neither periods of perceived hunger nor illness were associated to differences in stature. Ethanol consumption during growth showed a significant association with the presence of Harris lines (chi(2)=15, P<.001, Odds Ratio [OR]=3.39, confidence interval [CI]=1.81-6.33), an association which was more marked between having two or more Harris lines and drinking during growth (chi(2)=23.19, P<.001, OR=6.04, CI=2.79-13.11) or having three or more lines and drinking during growth (chi(2)=15.93, P<.001, OR=7.41, CI=2.47-22.21). Periods of perceived hunger during growth were also related to the presence of two or more Harris lines (chi(2)=4.66, P=.031, OR=2.055, CI=1.065-3.965), but no association was observed between illness and Harris lines, two or more Harris lines, and three or more Harris lines. Multivariate analysis showed that only ethanol consumption during growth period was associated with Harris lines. PMID- 17913442 TI - Deep proteome profiling of sera from never-smoked lung cancer patients. AB - Previous studies on the serum proteome are hampered by the huge dynamic range of concentration of different protein species. The use of Equalizer Beads coupled with a combinatorial library of ligands has been shown to allow access to many low-abundance proteins or polypeptides undetectable by classical analytical methods. This study focused on never-smoked lung cancer, which is considered to be more homogeneous and distinct from smoking-related cases both clinically and biologically. Serum samples obtained from 42 never-smoked lung cancer patients (28 patients with active untreated disease and 14 patients with tumor resected) were compared with those from 30 normal control subjects using the pioneering Equalizer Beads technology followed by subsequent analysis by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Eighty-five biomarkers were significantly different between lung cancer and normal control. The application of classification algorithms based on significant biomarkers achieved good accuracy of 91.7%, 80% and 87.5% in class-prediction with respect to presence or absence of disease, subsequent development of metastasis and length of survival (longer or shorter than median) respectively. Support vector machine (SVM) performed best overall. We have proved the feasibility and convenience of using the Equalizer Beads technology to study the deep proteome of the sera of lung cancer patients in a rapid and high-throughput fashion, and which enables detection of low abundance polypeptides/proteins biomarkers. Coupling with classification algorithms, the technologies will be clinically useful for diagnosis and prediction of prognosis in lung cancer. PMID- 17913443 TI - Paclitaxel albumin-bound particles (abraxane) in combination with bevacizumab with or without gemcitabine: early experience at the University of Miami/Braman Family Breast Cancer Institute. AB - BACKGROUND: Paclitaxel albumin-bound particles (nab-paclitaxel, ABRAXANE) (nab-P) improve outcomes when compared against single agent cremophor-based paclitaxel, as do the addition of bevacizumab (B) or gemcitabine (G) to the same agent. There are no available data regarding combinations of nab-P with B and/or G. Ongoing investigational efforts are evaluating various doublets with these agents, but, to the best of our knowledge, not all 3 of them in the same regimen. All drugs are currently FDA-approved in the treatment of cancer. METHODS: Review of single institution experience, evaluating safety and preliminary evidence of activity with the use of nab-P and B with and without G in heavily pretreated her2neu negative metastatic breast cancer patients. Assessment of response was undertaken by the investigators independently of treating physician. RECIST criteria were used. RESULTS: Six women have been evaluated. Three patients received nab-P and B at the following doses: nab-P 100mg/m2, B 10mg/kg and 3 patients also received G at 1000 mg/m2; all 3 drugs were given every 2 weeks. Median age was 51 (range, 34 69). Two patients had hormone-receptor positive disease and 3 had ER/PR/her2neu negative cancer. Median prior number of regimens was 3 (range, 2-7). Five patients had been previously treated with a taxane. One received both paclitaxel and docetaxel, and 4 received docetaxel only. A median of 16 weeks of treatment has been administered (range 8+-32+). First-cycle grade 3/4 toxicity was seen in only one patient who had a baseline grade 2 thrombocytopenia that progressed to grade 3. The thrombocytopenia resolved without transfusion or hemorrhagic complication. Other treatment related toxicities were as follows: grade 2 peripheral neuropathy, 1 patient; grade 2 nausea, 1 patient. One patient had a blood pressure of 210/140 mmHg while non-compliant with her prior anti hypertensive therapy. Two patients had confirmed partial responses and 4 patients had stable disease. CONCLUSION: These very preliminary data suggest that nab-P in combination with B with and without G is a safe regimen and a formal phase II trial has been developed at the University of Miami to confirm its safety and clinical activity. PMID- 17913444 TI - Potentially useful biomarkers for the diagnosis, treatment and prognosis of lung cancer. AB - Lung cancer ranks top in both incidence and mortality in most part of the world. Scientists strive to explore biomarkers and their possible role in the diagnosis, treatment and prognosis of lung cancer. The ultimate goal is to discover biomarkers that can be tested in clinical trials and finally applied to patient care. Highly elevated concentrations of cytokeratin 19 fragment, tissue polypeptide antigen and squamous cell carcinoma antigen in non-small cell lung cancer particularly for squamous cell carcinoma, carcinoembryonic antigen and cancer antigen 125 in adenocarcinoma or non-small cell lung cancer, as well as progastrin-releasing peptide and neuron specific enolase in small cell lung cancer are suggestive biomarkers for the malignancy. Despite extensive studies, most results still remain controversial. Even with the report of high percent sensitivity and specificity, validation by clinical trials in large cohorts of patients is necessary before the cancer-related phenotypes can be translated into the clinic as reliable biomarkers. Nevertheless, identifications of biomarkers are leading to more understanding of the molecular pathways involved in lung cancer. It is hoped that understanding the connections between cellular pathways will help to reduce the suffering and loss of life caused by the lethal disease. This article summarizes the pre-clinical and translational researches against lung cancer in relation to biomarker discovery and validation. It is intended for policy makers, researchers, clinicians and other health professionals, offering a variety of useful biomarkers and updated data of clinical trials for lung cancer. PMID- 17913445 TI - Induction of matrix metalloproteinases by Epstein-Barr virus latent membrane protein 1 isolated from nasopharyngeal carcinoma. AB - Epstein--Barr virus latent infection is associated with human malignancies including Burkitt's lymphoma, gastric carcinoma and the highly invasive nasopharyngeal carcinoma (NPC). Increased expression of EBV latent membrane protein 1, LMP1, is correlated with tumor progression and metastasis in NPC. LMP1 induces cellular proteins including cytokines and matrix metalloproteinases (e.g., MMP1, MMP2 and MMP9). MMPs are endopeptidases involved in the degradation of extracellular matrix proteins; and their upregulation in cancer implicates their potential role in tumor metastasis. In light of the role of LMP1 in cytokine dysregulation and the fact that MMPs are regulated by cytokines, we examined whether LMP1 promotes NPC metastasis via the induction of MMPs. To delineate the oncogenic role of LMP1 in NPC, we first investigated the induction of MMP1, MMP2, MMP3 and MMP9 in LMP1-positive NPC tumor samples (n=15) by quantitative RT-PCR. We showed a significant induction of MMP1 and MMP3 transcripts in the EBV LMP1-positive NPC tissues, compared with biopsies obtained from the adjacent non-tumor tissues. To investigate the role of LMP1 in MMP expression in NPC, we cloned the LMP1 gene from NPC samples and transiently expressed it in MRC5 cells (human lung fibroblasts). Following transfection, a time-dependent elevation of endogenous MMP3 expression was found in the LMP1 transfectants by quantitative RT-PCR and Western analysis. Taken together, we observed that MMP3 is upregulated in LMP1-positive NPC tumors and LMP1-expression in fibroblasts is associated with MMP3 and cytokine expression. Our results suggest that LMP1 may contribute to invasiveness of NPC cells via the expression of MMP3 in fibroblasts. PMID- 17913446 TI - Effects of Shugansanjie Tang on matrix metalloproteinases 1, 3 and 9 and telomerase reverse transcriptase expression in human breast cells in vitro. AB - BACKGROUND: The traditional Chinese medication Shugansanjie Tang (SGT), whose active component is Akebia Trifoliate Koidz, possesses potential anti-tumor and immunostimulatory effects especially for breast cancer. The invasive processes of such cancers have been attributed to matrix metalloproteinases (MMPs) which may be the key factor mediating this process. Telomerase reverse transcriptase (TERT) catalyze the lengthening of telomeres, which prolongs cell life and interrupts natural cell death. The aim of this study is to determine the effects of Shugansanjie Tang on MMP levels and TERT activity using breast cancer cell lines. MATERIALS AND METHODS: We used the breast cancer cell lines MDA-MB-231 and BT 483. Cell inhibition rate was measured by WST-1 reagent, cell apoptosis by Apoptotic DNA Ladder Kit, mRNA expression of MMP-1, MMP-3 and MMP-9 by PCR and TERT by immunohistochemistry stain. RESULTS: Comparing to the control group, the test group showed lower cell growth rate, decreased mRNA expression of MMP-1, MMP 3 and MMP-9 production and less intense staining of MMPs with diaminobenzidine tetrahydrochloride. CONCLUSION: This study demonstrated that Shugansanjie Tang inhibits the growth of breast cancer cells by apoptosis and lowers the level of certain matrix metalloproteinases and activity of telomerase reverse transcriptase in breast cancer in vitro. PMID- 17913447 TI - Peripheral blood mononuclear cell NF-kappaB p105 mRNA decreases during asthmatic attacks. AB - BACKGROUND: NF-kappaB is a transcription factor involved in expression of many inflammatory cytokines, chemical transmitters, and adhesion molecules. It has been reported to play a major role in the pathogenesis of asthma. NF-kappaB p50, which is the actual subunit that results from the cleavage of p105, is required for the induction of eosinophilia via IL-5 and chemokines. METHODS: The subjects were 10 patients with a mean age of 59.3 years (14-82 years). NF-kappaB p105 mRNA in peripheral blood mononuclear cells during the presence or absence of asthmatic attacks was investigated. Total RNA was extracted from peripheral blood mononuclear cells. After cDNA was synthesized using random primers, NF-kappaB p105 mRNA level was measured by real-time polymerase chain reaction. RESULTS: The NF-kappaB p105 mRNA level in peripheral blood mononuclear cells was lower during asthmatic attacks than in the absence of attacks, showing a significant difference (Wilcoxon's signed rank test: p<0.01). CONCLUSIONS: A drop in NF kappaB p105 during an asthma attack could result in increased NF-kappaB activity. There is a possibility that a change in the NF-kappaB p105 mRNA level might indicate some pathogenetic state in bronchial asthma attacks. PMID- 17913448 TI - Neoadjuvant chemotherapy for breast cancer determined by chemosensitivity assay achieves better tumor response. AB - BACKGROUND: Neoadjuvant chemotherapy can potentially reduce tumor size and help downstage the tumor before definitive operation was performed. However, it was not possible to tell whether the patient would respond to the regimen until given. This difficulty can be overcome by testing the susceptibility of a sample of cancer cells in vitro: a "patient-tailored approach". In this pilot study, we attempt to demonstrate an improved response by this "patient-tailored" approach over standard regimen. MATERIALS AND METHODS: The study included 36 women with moderately advanced local breast cancer larger than 2 cm in diameter. Twelve were allocated to receive a standard regimen of 5-fluorouracil, epirubicin and cyclophosphamide (FEC) preoperatively as controls, and 24 were given the most suitable regimen according to testing; the options were FEC, cyclophosphamide, methotrexate and 5-fluorouracil (CMF), 5-fluorouracil, adriamycin and mitomycin C (FAM) and paclitaxel alone. The cell activities of drug-treated solid tumors were compared to controls with a highly sensitive ATP bioluminescence assay. Patients received chemotherapy according to sensitivity results and the tumor area clinically measured before and after chemotherapy. RESULTS: Sensitivity-directed treatment helped patients achieve a higher rate of complete clinical response (10/24 vs. 0/12), larger mean reduction in tumor area (75% vs. 26%), and 25% pathological complete response (pCR). The paclitaxel subgroup achieved 80% (pCR). CONCLUSION: It is a useful in vitro assay to provide a reference of the particular patient who received treatment according to her sensitivity result. It may improve pathologic complete response, clinical tumor response and lead to less extensive surgery. PMID- 17913449 TI - Fructus Schisandrae (Wuweizi) containing compound in modulating human lymphatic system - a Phase I minimization clinical trial. AB - BACKGROUND: Hepatitis B virus (HBV) infection afflicts Asia population and, in Hong Kong, about 10% was Hepatitis B surface antigen carrier. It is still one of the major issues under investigation. Herbal medicine KY88 composed of Fructus Schisandrae possessing immunomodulatory property was adopted by Chinese medicine practitioner for treatment of acute and chronic HBV infection. However, the underlying impact on host immune system is not fully understood. MATERIALS AND METHODS: Twenty-three healthy volunteers infected with HBV were taken peripheral venous blood from which the blood cells involved in simple host immunity was obtained. RESULTS: It was found that the circulating monocyte count significantly drop after 2weeks of KY88 therapy whereas the fall did not return back to baseline. Circulating white blood cell, neutrophil and lymphocyte, however, did not show obvious change upon commencement of KY88 therapy. CONCLUSION: It was postulated that reduction in circulating monocyte count may reduce the self inflicted host immune injury to hepatocyte which may testify the hepatoprotective ability of the herb. But, the exact mechanism on how immunomodulatory properties of the herbal medicine protect chronic HBV carriers from liver injury remains a myth. PMID- 17913450 TI - Balamuthia amoebic encephalitis: an emerging disease with fatal consequences. AB - Balamuthia amoebic encephalitis (BAE), caused by the protozoan pathogen, Balamuthia mandrillaris, is a serious human disease with fatal consequences and a mortality rate of more than 95%. A key factor that contributes to the high mortality is the incomplete understanding of its pathogenesis and pathophysiology. The most distressing aspect is that the high level of mortality is due to lack of awareness combined with the lack of effective drugs. Early diagnosis followed by aggressive treatment may lead to cure. Several lines of evidence suggest that BAE develops as a result of haematogenous spread, but it is unclear how circulating amoebae enter the central nervous system and cause inflammation, blood-brain barrier disruption and neuronal injury. Recent studies have identified several parasite-host determinants for B. mandrillaris translocation of the blood-brain barrier, and host inflammatory markers that may be associated with neuronal injury. These determinants may provide important targets for the prevention and treatment of BAE. Here, we present a brief overview of the current understanding of the pathogenesis and pathophysiology of BAE, available diagnostic methods, possible therapeutic interventions and biology of the causative agent. PMID- 17913451 TI - Hand skill assessment with a reduced version of the Peg Moving Task (PMT-5) in children: normative data and application in children with cerebral palsy. AB - This paper proposes a simple method of hand skill assessment in children that can be useful in clinical practice. A reduced 5-hole version of Annett's Peg Moving Task was used to quantify hand skill bilaterally in 435 normally developing preschool and school-children, and adolescents aged 3-18 years from Brazil. The cross-cultural validity of the normative data obtained in Brazil was verified in 157 school-children aged 6-11 years from France. An application in 76 children with cerebral palsy (hemiplegia 21, diplegia 34, triplegia 6, mixed type 15) showed very important variability of the deficits in hand function within each subtype of cerebral palsy (CP). Hand deficits were more severe in children in special schools than in children in regular schools within each CP subtype. A qualitative analysis showed which difficulties during the execution of the task were specific to children with CP and which were also observed in normally developing children. PMID- 17913452 TI - Confabulations in remembering past and planning future are associated with psychiatric symptoms in Alzheimer's disease. AB - Psychiatric symptoms such as delusions and aggression are frequently observed in patients with Alzheimer's disease (AD), but few studies examined the association of these symptoms with confabulations. We studied 32 AD patients and 10 age- and education-matched healthy older adults. The AD patients were divided into delusion/aggression and non-delusion/non-aggression groups based on their behavioral pathology in AD frequency-weighted severity scale score. Confabulations were assessed using questions about temporality (personal past, orientation, and future planning), and cognitive functions were determined using the mini-mental state examination and the cognitive abilities screening instrument. The AD patients showed confabulations on all types of questions, and their confabulation scores for the past and future were strongly correlated. Cognitive functions were not significantly correlated with confabulation scores for any type of questions. The delusion/aggression group had significantly more confabulations on past and future questions compared to the non-delusion/non aggression group. These findings suggested that confabulations in remembering the past and planning the future were affected by psychiatric symptoms such as delusion and aggression. PMID- 17913453 TI - Discriminative delay Pavlovian eyeblink conditioning in veterans with and without posttraumatic stress disorder. AB - BACKGROUND: Impaired eyeblink (EB) classical conditioning using a delay paradigm has previously been shown in combat veterans, as well as in a group of depressed adults, compared to normal individuals. Significant deficits in immediate memory (IM) in combat PTSD+ veterans, compared to normal controls, have also been previously shown, but these differences became non-significant after controlling for level of self-reported depression. Furthermore, EB conditioning has been shown to be significantly correlated with heart rate variability (HRV) in normal adults. The present study examined how depression (self-reported), IM, and resting HRV are related to discriminative delay classical EB conditioning in veterans with and without PTSD. METHOD: Three groups of subjects (combat PTSD+, combat PTSD-, and non-combat PTSD-) were assessed for self-report of depression and anxiety, as well as IM and HRV. Subjects received a single session of discriminative EB classical conditioning in which the conditioned stimulus (CS) was a light signal (either red or green) compounded with a tone. On CS+ trials, the light-tone compound stimulus co-terminated with a corneal airpuff (unconditioned stimulus, US), thus producing a delay paradigm. On CS- trials the appropriate light-tone stimulus was presented but not followed by the airpuff US. EB amplitude and frequency were recorded. RESULTS: PTSD+ subjects had greater self-reported depression and anxiety scores than the two control groups, as well as lower scores on a measure of IM. However, the IM difference was not significant after the effects of self-reported depression and anxiety were controlled. EB CR amplitude was significantly greater to CS+ than CS- for all three groups. EB amplitude to both the US (airpuff) and the CS+ declined over trials, but was significantly lower in the combat PTSD+ group compared to the combined PTSD- groups. Subjects who reached an EB CR acquisition criterion had significantly greater scores on IM than those who did not reach criterion. Factor analysis of the entire data set revealed four factors corresponding to (1) self reported depression and anxiety, (2) IM, (3) HRV, and (4) EB amplitude. EB frequency was significantly predicted by IM and HRV. CONCLUSIONS: These data extend our previous results by showing deficits in EB conditioning among combat PTSD+ veterans that were associated with lower IM and resting HRV, but were not associated with self-report of depression. PMID- 17913454 TI - The Obsessive-Compulsive Inventory-Revised (OCI-R): validation of the German version in a sample of patients with OCD, anxiety disorders, and depressive disorders. AB - The OCI-R is a psychometrically sound and valid self-report scale measuring the major symptoms of OCD on six dimensions: Checking, Washing, Ordering, Hoarding, Obsessing, and Neutralizing. Information is needed on its ability to discriminate OCD from depression. In this study, reliability and convergent, divergent, and known-groups validity of an authorized German version were examined in 381 patients with OCD, other anxiety and depressive disorders. Confirmatory factor analyses replicated the original six-factor structure in each sample. Moreover, results indicated good convergent, divergent, and known-groups validity for the full scale and the subscales in each sample, only a slight construct overlap between OCD and depression, anxiety, pathological worry, and perfectionism, and the relationships of the subscales with obsessive-compulsive personality features supported its construct validity. Previous findings for the original scale were replicated and extended in a different cultural context. However, the domains Neutralizing and Obsessions need further development. PMID- 17913456 TI - Rapid detection of Atopobium vaginae and association with organisms implicated in bacterial vaginosis. AB - Atopobium vaginae, a fastidious, anaerobic, Gram-positive cocci-shaped bacterium that generates large quantities of lactic acid, is associated with bacterial vaginosis (BV). Published nucleic acid amplification tests for identifying A. vaginae are directed toward the 16S ribosomal DNA with suboptimal specificity and require isolation of the organism. Here, sequencing of an A. vaginae genomic library has led to the development of a highly specific and sensitive real-time PCR test for detection of A. vaginae directly from gynecological cervicovaginal swab samples. The real-time PCR did not cross-react with DNA extracted from other members of the Atopobium genus, species with closely related 16S ribosomal DNA, and a panel of 51 other human pathogens. The DNA extraction and PCR assembly were amenable to automation using Corbett Robotics X-tractor Gene and CAS-4200N liquid handling systems. The real-time PCR was used to analyze 96 cervicovaginal swab samples submitted to our clinical laboratory for detection of organisms associated with BV. Of those samples, 28 were positive for A. vaginae. Of the 28 positive samples, 23 were concomitant with Gardnerella vaginalis detection. These results suggest that further clinical study of the relationship of A. vaginae with G. vaginalis and the development of BV should be performed. PMID- 17913455 TI - Atypical structure and phylogenomic evolution of the new eutherian oocyte- and embryo-expressed KHDC1/DPPA5/ECAT1/OOEP gene family. AB - Several recent studies have shown that genes specifically expressed by the oocyte are subject to rapid evolution, in particular via gene duplication mechanisms. In the present work, we have focused our attention on a family of genes, specific to eutherian mammals, that are located in unstable genomic regions. We have identified two genes specifically expressed in the mouse oocyte: Khdc1a (KH homology domain containing 1a, also named Ndg1 for Nur 77 downstream gene 1, a target gene of the Nur77 orphan receptor), and another gene structurally related to Khdc1a that we have renamed Khdc1b. In this paper, we show that Khdc1a and Khdc1b belong to a family of several members including the so-called developmental pluripotency A5 (Dppa5) genes, the cat/dog oocyte expressed protein (cat OOEP and dog OOEP) genes, and the ES cell-associated transcript 1 (Ecat1) genes. These genes encode structurally related proteins that are characterized by an atypical RNA-binding KH domain and are specifically expressed in oocytes and/or embryonic stem cells. They are absent in fish, bird, and marsupial genomes and thus seem to have first appeared in eutherian mammals, in which they have evolved rapidly. They are located in a single syntenic region in all mammalian genomes studied, except in rodents, in which a synteny rupture due to a paracentric inversion has separated this gene family into two genomic regions and seems to be associated with increased instability in these regions. Overall, we have identified and characterized a novel family of oocyte and/or embryonic stem cell-specific genes encoding proteins that share an atypical KH RNA-binding domain and that have evolved rapidly since their emergence in eutherian mammalian genomes. PMID- 17913457 TI - RPW: a hybrid reverse prediction method for level of detail. AB - In this paper, we propose a way of using multiple domain analysis methodology to speed up the image rendering process. We first apply wavelet transform to the original image, and then compress the wavelets in the frequency domain using histogram equalization techniques. When rendering the image, we uncompress the wavelets and reverse predict the upper level images. This process continues until it reaches a certain criteria. We use two terms-total image size (TIS) and total loading time (TLT) to measure the performance of level of detail (LOD) in a network environment. We compare traditional image-based LOD methods with the new method we are proposing. Experiments show that the proposed method can reduce both TIS and TLT. The image rendering speed on a remote client is approximately 2.5 times faster than the common image compression methods. Applications such as remote diagnostic systems and online museums can use this technique to achieve better real-time animation effects. PMID- 17913458 TI - Fractal research of pathological tissue images. AB - A novel texture detecting analysis for medical pathological tissue images was developed by fractal Brown model. According to fractal Brown random field model, a discrete fractal random field for image texture detection on a definite scale could be derived from the Brown model. Using fractal dimensions in partial region of the image and gray difference between adjacent pixels and relevant Hurst coefficient, a new medical image texture detecting analysis that could reflect the image feature was brought forward. The detecting results had been acquired. The results indicate that the texture detecting method by the Brown model is remarkable. PMID- 17913459 TI - Structural domains determining signalling characteristics of the CRH-receptor type 1 variant R1beta and response to PKC phosphorylation. AB - Mammalian adaptive mechanisms to stressful stimuli involve release of corticotropin-releasing hormone (CRH) and downstream activation of specific G protein-coupled 7 transmembrane domain receptors. These CRH receptors (CRH-R) are expressed as multiple mRNA spliced variants. In contrast to other mammals, the human type 1 CRH-R gene contains an additional exon (exon 6) that needs to be spliced out in order to generate the fully active CRH-R1alpha. Transcription of all 14 exons results in a CRH-R1 variant (CRH-R1beta) with an extended 1st intracellular loop (IC1); this sequence modification impairs signalling activity and alters receptor responsiveness to PKC-induced phosphorylation that leads to signalling desensitization and receptor endocytosis. To elucidate structure function relationships and delineate sequences involved in CRH-R1beta properties, site directed mutagenesis was used to introduce a number of specific mutations into IC1 of CRH-R1beta as well as replace specific phospho-acceptor residues within the aminoacid sequence of CRH-R1alpha and CRH-R1beta. Mutant receptors were transiently expressed in human embryonic kidney (HEK293) cells and tested for their abilities to increase intracellular cAMP and their response to PKC induced phosphorylation. Results identified a penta-aminoacid cassette within the 29-aminoacid insert of CRH-R1beta, which contains multiple positive charged aminoacids (F170-R174), as an important structural determinant for the impaired cAMP response. Furthermore, serine at position 408 in the carboxy-terminus appears to be important for mediating CRH-R1alpha resistance, but not CRH-R1beta susceptibility, to PKC-induced desensitization and internalization. These findings provide new insights about the structural determinants of CRH-R1 coupling to Gs proteins and response to protein kinase phosphorylation. PMID- 17913460 TI - A case of tumoural calcinosis in the temporomandibular joint associated with systemic sclerosis. AB - Systemic sclerosis (SSc) is a relatively rare condition characterized by the excessive production and deposition of collagen within tissue. This condition is thought to be immunologically mediated and, in addition to its notorious cutaneous manifestations, often involves multiple organs. A case is presented of systemic sclerosis associated with extensive tumoural calcinosis in the temporomandibular joint. There has been no evidence of recurrence or complications during approximately 2 years of follow up, but long-term follow up is essential. PMID- 17913461 TI - Factors influencing the prophylactic removal of asymptomatic impacted lower third molars. AB - The aim of this study was to analyse factors indicating prophylactic removal of impacted lower third molars, and how they, and possibly surgeon experience, influence the therapeutic decision-making process. A descriptive observational study was made of 40 asymptomatic impacted lower third molars. Orthopantomographs were scanned and presented to four professionals with different degrees of surgical experience. The examiners received information relating to patient age and sex, molar inclination and degree of impaction, and expressed their opinion on the necessity for teeth removal. There was a statistically significant relationship between examiner decision and the estimated probability of pathology if the molars were not removed (P<0.05). The degree of influence on the decision to extract was in decreasing order: estimated risk of complications, inclination of molar, age, degree of impaction and patient sex. No statistically significant differences (P>0.05) were observed between residents and trainers in terms of the decision to remove or estimated risk of complications. The management approach adopted by oral surgeons regarding the removal of asymptomatic impacted lower third molars depends upon the perceived risk of complications if such teeth are not removed, other factors being secondary. The surgical experience of the professional does not seem to influence treatment decision. PMID- 17913462 TI - The controversy continues!! PMID- 17913463 TI - Parotid metastasis from carcinoma of the thyroid. PMID- 17913464 TI - Effects of nociceptin on the spread and seizure activity in the rat amygdala kindling model: their correlations with 3H-leucyl-nociceptin binding. AB - The effects with pretreatment with nociceptin (0.03-30nmol, i.c.v.) were evaluated on the threshold for eliciting afterdischarge (ADT), generation and spread of seizure activity and postictal depression in rats with kindling stimulation. Nociceptin produced a decrease in ADT (32-45%) in rats with partial seizures (PS, stage II-III), and an increase (61-92%) in rats with generalized seizures (GS, kindled state). Nociceptin did not modify the behavioral changes, spike frequency and duration of afterdischarge elicited at ADT in both experimental groups. In rats with GS, nociceptin enhanced postictal depression (34-44%) evaluated with a recycling paradigm. Autoradiography experiments revealed enhanced nociceptin opioid receptor (NOP) binding in medial amygdala (22 26%), frontal (21-23%) and entorhinal (27-32%) cortices, and reduced binding in the substantia nigra pars compacta (28%) and medial central gray (29%) of rats with PS. The GS group displayed significant decreased NOP binding (40-70%) in most of the brain areas evaluated. These results suggest that nociceptin facilitates ictal activity in rats with PS, whereas in animals with GS, it induces inhibitory effects on ADT and enhances the postictal period. These effects correlate with significant changes in NOP binding. PMID- 17913465 TI - Volumetric abnormalities in connectivity-based subregions of the thalamus in patients with chronic schizophrenia. AB - OBJECTIVE: The thalamus, which consists of multiple subnuclei, has been of particular interest in the study of schizophrenia. This study aimed to identify abnormalities in the connectivity-based subregions of the thalamus in patients with schizophrenia. METHODS: Thalamic volume was measured by a manual tracing on superimposed images of T1-weighted and diffusion tensor images in 30 patients with schizophrenia and 22 normal volunteers. Cortical regional volumes automatically measured by a surface-based approach and thalamic subregional volumes measured by a connectivity-based technique were compared between the two groups and their correlations between the connected regions were calculated in each group. RESULTS: Volume reduction was observed in the bilateral orbitofrontal cortices and the left cingulate gyrus on the cortical side, whereas in subregions connected to the right orbitofrontal cortex and bilateral parietal cortices on the thalamic side. Significant volumetric correlations were identified between the right dorsal prefrontal cortex and its related thalamic subregion and between the left parietal cortex and its related thalamic subregion only in the normal group. CONCLUSIONS: Our results suggest that patients with schizophrenia have a structural deficit in the corticothalamic systems, especially in the orbitofrontal-thalamic system. Our findings may present evidence of corticothalamic connection problems in schizophrenia. PMID- 17913466 TI - Heart rate variability response to mental arithmetic stress in patients with schizophrenia: autonomic response to stress in schizophrenia. AB - BACKGROUND: The vulnerability-stress hypothesis is an established model of schizophrenia symptom formation. We sought to characterise the pattern of the cardiac autonomic response to mental arithmetic stress in patients with stable schizophrenia. METHODS: We performed heart rate variability (HRV) analysis on recordings obtained before, during, and after a standard test of autonomic function involving mental stress in 25 patients with DSM-IV schizophrenia (S) and 25 healthy individuals (C). RESULTS: Patients with schizophrenia had a normal response to the mental arithmetic stress test. Relative contributions of low frequency (LF) HRV and high-frequency (HF) HRV influences on heart rate in patients were similar to controls both at rest (LF 64+/-19% (S) vs. 56+/-16% (C); HF 36+/-19% (S) vs. 44+/-16% (C), t=1.52, p=0.136) and during mental stress, with increased LF (S: 76+/-12%, C: 74+/-11%) and decreased HF (S: 24+/-12%, C: 26+/ 11%) in the latter study condition. Whilst healthy persons recovered the resting pattern of HRV immediately after stress termination (LF 60+/-15%, HF 40+/-15%, F=18.5, p<0.001), in patients HRV remained unchanged throughout the observed recovery period, with larger LF (71+/-17%) and lower HF (29+/-17%) compared with baseline (F=7.3, p=0.013). CONCLUSIONS: Patients with schizophrenia exhibit a normal response to the mental arithmetic stress test as a standard test of autonomic function but in contrast with healthy individuals, they maintain stress related changes of cardiac autonomic function beyond stimulus cessation. PMID- 17913467 TI - The ancestral role of the phosphoenolpyruvate-carbohydrate phosphotransferase system (PTS) as exposed by comparative genomics. AB - The normal role of the phosphoenolpyruvate-carbohydrate phosphotransferase system (PTS) is phosphorylation and subsequent uptake of specific sugars. However, analysis of the distribution of PTS proteins in 206 genomes covering major bacterial groups indicates that the conventional function of PTS proteins as devices for carbohydrate phosphorylation and transport is an exception found in Enterobacteriacea, Vibrionales and Firmicutes, rather than a rule for all bacteria. Instead, available evidence suggests that a core set of C-responsive phosphotransferases have been evolutionarily drafted towards diversity of regulatory functions in response inter alia to the global economy of the C and N pools. PMID- 17913468 TI - Current challenges in autotransport and two-partner protein secretion pathways. AB - The two-partner and autotransport pathways are widely represented in Gram negative bacteria, essentially among pathogens. Both mediate the translocation of large proteins across the outer membrane. Deciphering the molecular mechanisms of secretion machineries as well as folding of exoproteins in the course of translocation represents a current challenge. PMID- 17913469 TI - Characterisation of plasmids encoding extended-spectrum beta-lactamase and CMY-2 in Escherichia coli isolated from animal farms. PMID- 17913470 TI - Tuberculosis in patients with systemic rheumatic or pulmonary diseases treated with glucocorticosteroids and the preventive role of isoniazid: a review of the available evidence. AB - Development of tuberculosis (TB) is a concern in patients who receive glucocorticosteroids for the treatment of chronic rheumatic or pulmonary diseases. However, the incidence of development of TB in such patients and the prophylactic role of isoniazid (INH) are unclear. We evaluated the available evidence from 20 relevant prospective and retrospective cohort and case-control studies identified in the PubMed and Cochrane databases. The frequency of development of TB in the populations studied varied from 0% to 13.8%. This figure was low in studies performed in countries with a low incidence of TB (0% in the USA and Greece, 0.6% in France and 1.35% in Spain). In contrast, the frequency of development of TB in the studied cohorts was high in studies performed in countries with a moderate to high incidence of TB (from 2.5% in South Korea to 13.8% in The Philippines). Isoniazid prophylaxis (INHP) was found to decrease the incidence of development of TB in two of four studies that examined this intervention. The available evidence suggests that patients who receive steroids for the treatment of chronic rheumatic or pulmonary diseases and who live in countries with a high incidence of TB have a high risk of development of TB in contrast to patients who live in countries with a low incidence of the infection. However, the role of INHP even for patients living in countries where TB is endemic is unclear because the effectiveness of INH in preventing TB development in such patients is not well established and there are cost-effectiveness and safety issues. PMID- 17913471 TI - Immunostimulation with Escherichia coli extract: prevention of recurrent urinary tract infections. AB - Antibiotic treatment is the most effective method for managing recurrent uncomplicated urinary tract infections (UTIs) in women. However, there is a limitation to long-term antibiotic use, therefore alternative approaches are required. Escherichia coli is the commonest cause of recurrent uncomplicated UTIs and accounts for more than 60% of recurrent cystitis. It has therefore been suggested that immunostimulation with a bacterial extract consisting of immunostimulating components derived from 18 E. coli strains may protect against UTIs. Many in vitro studies, animal experiments and clinical trials in patients with recurrent UTIs have been carried out to assess the effectiveness and safety of E. coli extract for prophylaxis against recurrent UTIs. In this paper, we review the scientific evidence showing the anti-inflammatory effect of E. coli extract as well as the clinical results of placebo-controlled, randomised, double blind studies that demonstrated a positive effect for E. coli extract in patients with recurrent UTIs. Evidence from available studies suggests that E. coli extract can be beneficial and safe for preventing recurrent UTIs in women. PMID- 17913472 TI - Metallo-beta-lactamase IMP-18 is located in a class 1 integron (In96) in a clinical isolate of Pseudomonas aeruginosa from Mexico. PMID- 17913473 TI - Phosphodiesterase 10A inhibition is associated with locomotor and cognitive deficits and increased anxiety in mice. AB - Phosphodiesterase 10A (PDE10A) mRNA and protein levels decline in the striatum of R6/1 and R6/2 Huntington's disease (HD) mice prior to motor symptom development. In human HD, PDE10A protein levels are significantly decreased in the caudate putamen of patients with grade 3 HD compared to age-matched controls. To test whether the loss of PDE10A activity in the striatum was detrimental to normal brain function, we treated wild-type (WT) mice with chronic administration of papaverine, which is a specific inhibitor of PDE10A. At 7 weeks of age, mice were introduced to a weekly battery of motor tests, including assessment of weight, locomotion, gait, and coordination. Beginning at 8 weeks of age, mice received 0, 5, 10 or 20 mg/kg papaverine once daily until the completion of behavioral testing. Following 14 days of papaverine injections, mice were assessed for deficits in cognitive performance as measured in the Morris water maze (MWM). All behavioral tests occurred either immediately prior to or 30 min following a subcutaneous papaverine challenge dose. Twenty-four hours following completion of the 2-3 week MWM protocol, mice were given a dose of papaverine and 30 min later psychological function assessed in the Light-Dark (LD) Test. Chronic administration of papaverine for 42 days was associated with distinct motor perturbations, mild cognitive disturbance and anxiety-like behaviors. Subsequently, we assessed the effect of 14 days papaverine (i.e. sub-chronic) treatment on psychological function of WT and R6/1 HD mice. While sub-chronic papaverine induced anxiety-like behavior in WT mice, it appeared to have little effect on the behavior of R6/1 HD mice. Finally, a separate group of 6-week old WT and R6/2 HD mice were treated for 21 days with saline or 10 mg/kg fluoxetine, an agent with anxiolytic and anti-depressant effects, in order to compare the effects of papaverine and fluoxetine on anxiety-like behavior in the LD test. CREB and PDE10A protein levels in striatum and hippocampus were determined by western blot. While papaverine treatment reduced CREB protein levels in the hippocampus and striatum, fluoxetine increased CREB in the hippocampus. These data suggest that papaverine and fluoxetine may produce quite different effects on behavior; these behaviors may be linked to CREB expression in brain regions associated with motor and cognitive functions. PDE10A protein levels were decreased by both papaverine and fluoxetine. Chronic PDE10A inhibition produced a variety of behavioral and central neurochemical deficits and these effects were exacerbated by stress. The unique localization of PDE10A and its apparent role in basal ganglia function may underlie its role in psychiatric and neurological disorders involving the basal ganglia. PMID- 17913474 TI - Atypical involvement of frontostriatal systems during sensorimotor control in autism. AB - Autism is a neurodevelopmental disorder involving dysmaturation of widely distributed brain systems. Accordingly, behaviors that depend on distributed systems, such as higher level cognition and sensorimotor control, are compromised in the disorder. The current study investigated alterations in neural systems underlying sensorimotor disturbances in autism. An fMRI investigation was conducted using saccadic and pursuit eye movement paradigms with 13 high functioning individuals with autism and 14 age- and IQ-matched typically developing individuals. Individuals with autism had reduced activation in cortical eye fields and cerebellar hemispheres during both eye movement tasks. When executing visually guided saccades, individuals with autism had greater activation bilaterally in a frontostriatal circuit including dorsolateral prefrontal cortex, caudate nucleus, medial thalamus, anterior and posterior cingulate cortex, and right dentate nucleus. The increased activation in prefrontal-striatal-thalamocortical circuitry during visually guided saccades indicates that systems typically dedicated to cognitive control may need to compensate for disturbances in lower-level sensorimotor systems. Reduced activation throughout visual sensorimotor systems may contribute to saccadic and pursuit disturbances that have been reported in autism. These findings document that neurodevelopmental disturbances in autism affect widely distributed brain systems beyond those mediating language and social cognition. PMID- 17913476 TI - Adsorption of concanavalin A and lentil lectin on platinum electrodes followed by electrochemical impedance spectroscopy: effect of protein state. AB - Adsorption of concanavalin A and lentil lectin on platinum electrode was investigated through electrochemical impedance spectroscopy and cyclic voltammetry. By using ferro/ferricyanide system to probe the electrochemical interface it was possible to model the EIS data with a simple equivalent circuit. The blocking effect for electron transfer reactions observed with these proteins, indicated that they readily adsorb on platinum surface and that the degree of adsorption is related to the state of the proteins. When the proteins are in the presence of divalent cations (Ca(2+) and Mn(2+)) they adsorb less strongly than in their absence. There is also evidence that at least convanavalin A retains its biological activity in the adsorbed state. PMID- 17913475 TI - Estimation of the specific surface area of apatites in human mineralized tissues using 31P MAS NMR. AB - Specific surface areas of apatites in whole human mineralized tissues were estimated from (31)P MAS NMR linewidths: 77 m(2)g(-1) for enamel and 94 m(2)g(-1) for dentin, dental cementum and cortical bone. PMID- 17913477 TI - Hepatocytes are protected by herb Phyllanthus niruri protein isolate against thioacetamide toxicity. AB - The herb, Phyllanthus niruri has been known to possess protective activity against various drugs and toxins induced hepatic disorders. Present study was conducted to evaluate the role of the protein isolate of the herb against thioacetamide (TAA)-induced cytotoxicity in mice hepatocytes. In vitro cell viability, lactate dehydrogenase (LDH) and alanine amino transferase (ALT) leakage were measured as the indicators of cell damage. In addition, measurement of the level of non-protein thiol, glutathione (GSH); activities of the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione S transferase (GST) as well as the extent of lipid peroxidation were carried out to evaluate the prooxidant-antioxidant status of the cell. 2,2-Diphenyl-1 picrylhydrazyl (DPPH) radical scavenging assay was performed to determine the radical scavenging activity of the protein isolate. Results showed that the administration of the protein isolate prior to TAA exposure significantly reduced the release of LDH and ALT leakage and enhanced the cell viability in a dose dependent manner in hepatocytes. Besides, the protein isolate appeared to prevent the alterations in GSH levels and activities of the anti-oxidant enzymes related to prooxidant-antioxidant status of hepatocytes. It also reduced the TAA-induced lipid peroxidation significantly as demonstrated by the reduction of malondialdehyde (MDA) production. DPPH radical scavenging assay showed that the protein isolate possessed radical scavenging activity. Combining, the data suggest that the protein isolate could protect hepatocytes from TAA-induced cellular injury probably by its antioxidative and radical scavenging properties. PMID- 17913478 TI - Daily running promotes spatial learning and memory in rats. AB - Previous studies on exercise have shown that physical activity improves learning and memory. Present study was performed to determine the effects of acute, chronic and continuous exercise with different periods on spatial learning and memory recorded as the latency and length of swim path in the Morris water maze testing in subsequent 8 days. Four rat groups were included as follows: (1) group C, controls which did not exercise; (2) group A, 30 days treadmill running before and 8 days during the Morris water maze testing period; (3) group B, 30 days exercise before the Morris water maze testing period only; (4) group D, 8 days exercise only during the Morris water maze testing period. The results showed that chronic (30 days) and continuous (during 8 days of Morris water maze testing days) treadmill training produced a significant enhancement in spatial learning and memory which was indicated by decreases in path length and latency to reach the platform in the Morris water maze test (p<0.05). The benefits in these tests were lost in 3 days, if the daily running session was abandoned. In group D with acute treadmill running (8 days exercise only) the difference between the group A disappeared in 1 week and benefit seemed to be obtained in comparison with the controls without running program. In conclusion the chronic and daily running exercises promoted learning and memory in Morris water maze, but the benefits were lost in few days without daily running sessions in adult rats. PMID- 17913480 TI - Gene Ontology analysis in multiple gene clusters under multiple hypothesis testing framework. AB - OBJECTIVE: Gene Ontology (GO) has become a routine resource for functional analysis of gene lists. Although a number of tools have been provided to identify enriched GO terms in one or two gene lists, two technical challenges remain. First, how to handle multiple hypothesis testing in the analysis given that the tests are heavily correlated; second, how to identify GO terms that are enriched in a gene cluster, as compared to multiple other gene clusters. We provide a statistical procedure to rigorously treat these problems and offer a software tool for applying GO to the analysis of gene clusters. METHODS: We previously introduced a statistical procedure that handles hypothesis testing in a two-group comparison scenario. In this paper we extend the two-group comparison procedure into a general procedure that enables the analysis of any number of gene lists/clusters. This new procedure enables identification of GO terms enriched in any gene cluster, while it controls for multiple hypothesis testing. This procedure is implemented into a user-friendly analysis tool: GoSurfer. The current version of GoSurfer takes one or several gene lists as input, and it identifies the GO terms that are enriched in any of the input gene lists. GoSurfer estimates a conservative false discovery rate (FDR) for every GO term. The FDR estimation procedure in GoSurfer has two advantages: it does not rely on independence assumption, and it does not assume all the hypotheses are null hypothesis (complete null). Thus GoSurfer's FDR estimates are mildly conservative rather than overly conservative. RESULTS: We implemented the new procedure for GO analysis in multiple gene clusters into the GoSurfer software. We provide three examples on using GoSurfer to analyze time course gene expression data sets on the differentiation of embryonic stem cells. In the example of analysis of multiple gene clusters, we first used a typical clustering algorithm and identified five gene clusters, representing up-regulation, down-regulation and other patterns in the differentiation time course. Taking all the five gene clusters as input data, GoSurfer reports "cell adhesion" and "muscle contraction" as significant GO terms for the up-regulated cluster, "amino acids metabolism" as a significant GO term for the down-regulated gene cluster, and GoSurfer reports a number of GO terms related to RNA processing and RNA transport as significant terms to a cluster that is up-regulated in both early and late time points. This may suggest that genes for RNA processing and genes for RNA transport are coregulated in the differentiation process of embryonic stem cells. CONCLUSION: The GoSurfer software is provided to analyze multiple gene clusters and identify GO terms that are enriched in any gene cluster. Gosurfer is available at: www.gosurfer.org. PMID- 17913479 TI - Robust effects of Tsr-CheBp and CheA-CheYp affinity in bacterial chemotaxis. AB - OBJECTIVE: Cell motility and chemotaxis play a role in the virulence of pathogenic bacteria, such as escape from host immune responses. Escherichia coli chemotaxis provides a well-characterized model system for the bacterial chemotaxis network. Two features of E. coli chemotaxis include signal amplification and robustly accurate adaptation. Recent simulation studies with models considering the effects of other receptors have suggested possible mechanisms for signal amplification. Although precise adaptation to aspartate has been explained by conventional kinetic models, the adaptation behavior of models incorporating the effects of other receptors remains unclear. METHODS: We concentrated on how receptor crosstalk affects minimization of adaptation error and compared models in which the contribution of other receptors varied. RESULTS: We demonstrated that the model is adaptable to attractant concentrations ranging from 0.1microM to 10mM with a decreased error rate (from 8% to 2%) when the kinetic constant of CheA and phosphorylated CheY dissociation is increased. CONCLUSION: The results suggest that accurate adaptation is maintained through control of both the interaction of cytoplasmic Che proteins and the activity of the receptor complex. PMID- 17913481 TI - Ultrastructure and development of the rhabdomeric eyes in Lineus viridis (Heteronemertea, Nemertea). AB - Nemerteans are undoubtedly members of the Spiralia, although their phylogenetic relationships are still a matter of debate. The apparently acoelomate organization suggests a relationship with the platyhelminths, whereas the blood vascular system has been interpreted as an equivalent to coelomic cavities of annelids, indicating a close relation between annelids and nemerteans. Like other spiralians, most nemertean species are known to have one or several pairs of rhabdomeric and subepidermally situated eyes when adult. The development of these eyes as well as the mode in which the eyes are multiplied is as yet unknown. This is the first attempt to investigate eye formation in a nemertean. In the heteronemertean Lineus viridis (Muller, 1774) the everse rhabdomeric eyes are located deeply underneath the epidermis and consist of a few pigment cells that form a cup-like structure with interdigitating processes that contain numerous pigment granules. In hatchlings, the optical cavity contains processes of 12 sensory cells, each bearing a single cilium and various microvilli. The perikarya of these cells are located distally from the pigment cup. During further development the number of cells increases. Eye development starts with a small anlage situated underneath the epidermis, irrespective of whether this is the first eye or any additional one. The anlage consists of five unpigmented cells and three dendritic processes, each bearing apical microvilli and a single cilium. There is no evidence for an epidermal origin of the eyes. In L. viridis eye formation resembles that described in platyhelminths in which eyes also develop as cerebral derivatives. Although this result has the potential to influence the discussion on the position of Nemertea, the data have to be interpreted with care, since development of L. viridis is derived within the Nemertea. PMID- 17913482 TI - Antimalarial activity of anthothecol derived from Khaya anthotheca (Meliaceae). AB - Antimalarial activity of anthothecol, a limonoid of Khaya anthotheca (Meliaceae) against Plasmodium falciparum was tested using a [(3)H]-hypoxanthine and 48h culture assay in vitro. Anthotechol showed potent antimalarial activity against malaria parasites with IC(50) values of 1.4 and 0.17microM using two different assays. Also, gedunin had antimalarial activity with IC(50) values of 3.1 and 0.14microM. However, the citrus limonoids, limonin and obacunone did not show any antimalarial activity. The antimalarial activities were compared with the three currently used antimalarial medicines quinine, chloroquinine and artemisinin. PMID- 17913483 TI - Inhibitory effect of compounds from Zingiberaceae species on human platelet aggregation. AB - Twelve compounds isolated from Alpinia mutica Roxb., Kaempferia rotunda Linn., Curcuma xanthorhiza Roxb., Curcuma aromatica Valeton and Zingiber zerumbet Smith (Family: Zingiberaceae) and three synthesized derivatives of xanthorrhizol were evaluated for their ability to inhibit arachidonic acid- (AA), collagen- and ADP induced platelet aggregation in human whole blood. Antiplatelet activity of the compounds was measured in vitro by the Chrono Log whole blood aggregometer using an electrical impedance method. Among the compounds tested, curcumin from C. aromatica, cardamonin, pinocembrine and 5,6-dehydrokawain from A. mutica and 3 deacetylcrotepoxide from K. rotunda showed strong inhibition on platelet aggregation induced by AA with IC(50) values of less than 84 microM. Curcumin was the most effective antiplatelet compound as it inhibited AA-, collagen- and ADP induced platelet aggregation with IC(50) values of 37.5, 60.9 and 45.7 microM, respectively. PMID- 17913484 TI - Synergic effect of grape seed extract with amphotericin B against disseminated candidiasis due to Candida albicans. AB - Amphotericin B (Amp B) is considered as a drug of choice for treatment of fungal infections, but it causes severe side effects such as renal damage. To lessen the severity, it is often combined with the azole, but data reporting resistance of Candida albicans to the azole have been recently increasing. Thus, finding a new product that can reduce Amp B dose by combination seems to be important. In the present study, we investigated a synergic effect of grape seed extract (GSE) combined with Amp B against the fungus. Our results showed that the GSE alone inhibited growth of C. albicans yeast cells, and that in a murine model of disseminated candidiasis mice groups given GSE before intravenous inoculation with the yeast cells survived longer than diluent-received (control) mice groups (P<0.05). This GSE antifungal effect was dose-dependent. Upon combination of GSE plus Amp B, the combination therapy strikingly retarded the yeast growth as determined by the broth susceptibility method. Against the disseminated disease, mice given diluent (negative control), Amp B (0.5mg/kg of body weight), or GSE (2mg/kg of body weight) had mean survival times (MSTs) of approximately 11.4, 14.4, and 17.6 days, respectively. However, mice treated with the combination of the doses of Amp B and GSE had a MST value of 38.4 days, surviving an average of 24 days longer than Amp B alone-treated mice groups. This MST value from the combination-received mice group was greater than the MST value from the mice group given four times the Amp B dose (2mg/kg of body weight). All these data indicate that the combination therapy can reduce more than 75% of Amp B dose, implying that GSE has a synergic effect with Amp B. PMID- 17913485 TI - Antiproliferative activity of Pterodon pubescens Benth. seed oil and its active principle on human melanoma cells. AB - On a preliminary screening, relevant in vitro antiproliferative activity was observed to the crude ethanolic extract of Pterodon pubescens seed oil against the human melanoma cell line SK MEL 37. The diethyl ether fraction from crude ethanolic extract which exhibited stronger activity was submitted to fractionation by gradient elution with hexane/ethyl acetate. Subfraction A, eluted by hexane/ethyl acetate (80:20), was essentially the most active between all the assayed subfractions with an IC(50) of 37microg/ml calculated by the MTT colorimetric method. At this concentration, subfraction A caused morphological features and internucleosomal DNA fragmentation pattern of apoptosis. Through chromatographic separation, the furane diterpene 1 was isolated from this active subfraction and identified by spectral techniques. Compound 1 showed an IC(50) value of 32microM and fluorescence staining with DAPI revealed some typical nuclear changes which are characteristic of apoptosis. These findings support a role for diterpenoids vouacapan-type skeleton as a model to develop new anticancer agents. PMID- 17913486 TI - Dispensing of antibiotics, antitussives and mucolytics to asthma patients: a pharmacy-based observational survey. AB - BACKGROUND: Antibiotics, antitussives and mucolytics are commonly used in asthma, despite limited evidence for their effectiveness. The correlates of use for these medication classes in asthma were identified. METHODS: Asthma patients aged 18-50 who were regular customers of pharmacies were included in an observational study. Patients completed a questionnaire, which was complemented by computerised pharmacy records of previously dispensed medications. Users of each drug class were compared to non-users in terms of demographics, asthma characteristics and management. RESULTS: Among 886 patients (mean age: 37; 55% females), 63.2%, 55.8% and 27.2%, respectively, were users of antibiotics, mucolytics and antitussives during the previous 12 months. In multivariate analysis, dispensing of >2 units of oral corticosteroids was the major correlate of receiving antibiotics (OR=5.47; 95% CI=[3.00-9.97]), mucolytics (OR=3.93; 95% CI=[2.38-6.50]) and antitussives (OR=1.86; 95% CI=[1.18-2.94]). Compared to well-controlled patients, the probability of receiving antibiotics was significantly higher for poorly controlled patients (OR=2.01; 95% CI=[1.28-3.15]). CONCLUSIONS: Our results suggest that these drugs are mainly used during asthma exacerbations. A better understanding of the use of co-medication in asthma is required. PMID- 17913487 TI - Real-time monitoring of adhesion and aggregation of platelets using thickness shear mode (TSM) sensor. AB - Hemostasis is required to maintain vascular system integrity, but thrombosis, formation of a clot in a blood vessel, is one of the largest causes of morbidity and mortality in the industrialized world. Novel clinical and research tools for characterizing the hemostatic system are of continued interest, and the object of this research is to test the hypothesis that clinically relevant platelet function can be monitored using an electromechanical sensor. A piezoelectric thickness shear mode (TSM) biosensor coated with collagen-I fibers to promote platelet activation and adhesion was developed and tested for sensitivity to detect these primary events. Magnitude and frequency response of the sensor were monitored under static conditions at 37 degrees C, using platelet-rich plasma (PRP), and PRP with adenosine diphosphate (ADP), a clinical aggregation inhibitor (abciximab), or a collagen binding inhibitor. Sensors loaded with PRP exhibited a 3-stage response; no significant change in response for the first 20 min (Stage 1), followed by a larger drop in response (Stage-2) and subsequently, response gradually increased (Stage-3). Exogenous ADP stimulated an immediate Stage-2 response, while abciximab delayed and reduced the magnitude change of Stage-2. In the presence of collagen inhibitor, Stage-2 response was similar to that of control but was delayed by an additional 20 min. The obtained results, supported by epifluorescence and complementary SEM studies, demonstrated the selective sensitivity of TSM electromechanical biosensors to monitor platelet function and inhibition, particularly aggregation. PMID- 17913489 TI - Side effects of genome structural changes. AB - The first extensive catalog of structural human variation was recently released. It showed that large stretches of genomic DNA that vary considerably in copy number were extremely abundant. Thus it is conceivable that they play a major role in functional variation. Consistently, genomic insertions and deletions were shown to contribute to phenotypic differences by modifying not only the expression levels of genes within the aneuploid segments but also of normal copy number neighboring genes. In this report, we review the possible mechanisms behind this latter effect. PMID- 17913488 TI - We gather together: insulators and genome organization. AB - When placed between an enhancer and promoter, certain DNA sequence elements inhibit enhancer-stimulated gene expression. The best characterized of these enhancer-blocking insulators, gypsy in Drosophila and the CTCF-binding element in vertebrates and flies, stabilize contacts between distant genomic regulatory sites leading to the formation of loop domains. Current results show that CTCF mediates long-range contacts in the mouse beta-globin locus and at the Igf2/H19 imprinted locus. Recently described active chromatin hubs and transcription factories also involve long-range interactions; it is likely that CTCF interferes with their formation when acting as an insulator. The properties of CTCF, and its newly described genomic distribution, suggest that it may play an important role in large-scale nuclear architecture, perhaps mediated by the co-factors with which it interacts in vivo. PMID- 17913490 TI - The third dimension of gene regulation: organization of dynamic chromatin loopscape by SATB1. AB - Compartmentalized distribution of functional components is a hallmark of the eukaryotic nucleus. Technological advances in recent years have provided unprecedented insights into the role of chromatin organization and interactions of various structural-functional components toward gene regulation. SATB1, the global chromatin organizer and transcription factor, has emerged as a key factor integrating higher-order chromatin architecture with gene regulation. Studies in recent years have unraveled the role of SATB1 in organization of chromatin 'loopscape' and its dynamic nature in response to physiological stimuli. SATB1 organizes the MHC class-I locus into distinct chromatin loops by tethering MARs to nuclear matrix at fixed distances. Silencing of SATB1 mimics the effects of IFNgamma treatment on chromatin loop architecture of the MHC class-I locus and altered expression of genes within the locus. At genome-wide level, SATB1 seems to play a role in organization of the transcriptionally poised chromatin. PMID- 17913491 TI - Dynamic organization of the cell nucleus. AB - The dynamic organization of the cell nucleus into subcompartments with distinct biological activities represents an important regulatory layer for cell function. Recent studies provide new insights into the principles, by which nuclear organelles form. This process frequently occurs in a self-organizing manner leading to the assembly of stable but plastic structures from multiple relatively weak interaction forces. These can rearrange into different functional states in response to specific modifications of the constituting components or changes in the nuclear environment. PMID- 17913492 TI - Sensor complexes regulating two-component signal transduction. AB - Two-component signal transduction systems consisting of a sensor histidine kinase and a response regulator/transcription factor interpret a multitude of environmental and cellular signals and coordinate the expression of a wide array of genes in bacteria. Signal recognition by sensor histidine kinases is the province of a sensor complex consisting of several protein domains that together serve to augment or attenuate the activity of the histidine kinase and thereby of gene expression. Recent investigations have shown the diverse strategies bacteria use to assemble protein domains into the sensor complexes to accomplish signaling. Structural studies of such domains are leading to an understanding of the mechanisms by which sensor complexes recognize signals and regulate kinase activity. PMID- 17913494 TI - Electronic detectors for electron microscopy. AB - Due to the increasing popularity of electron cryo-microscopy (cryoEM) in the structural analysis of large biological molecules and macro-molecular complexes and the need for simple, rapid and efficient readout, there is a persuasive need for improved detectors. Commercial detectors, based on phosphor/fibre optics coupled CCDs, provide adequate performance for many applications, including electron diffraction. However, due to intrinsic light scattering within the phosphor, spatial resolution is limited. Careful measurements suggest that CCDs have superior performance at lower resolution while all agree that film is still superior at higher resolution. Consequently, new detectors are needed based on more direct detection, thus avoiding the intermediate light conversion step required for CCDs. Two types of direct detectors are discussed in this review. First, there are detectors based on hybrid technology employing a separate pixellated sensor and readout electronics connected with bump bonds-hybrid pixel detectors (HPDs). Second, there are detectors, which are monolithic in that sensor and readout are all in one plane (monolithic active pixel sensor, MAPS). Our discussion is centred on the main parameters of interest to cryoEM users, viz. detective quantum efficiency (DQE), resolution or modulation transfer function (MTF), robustness against radiation damage, speed of readout, signal-to noise ratio (SNR) and the number of independent pixels available for a given detector. PMID- 17913495 TI - Palliation of colorectal cancer. AB - Patients with advanced incurable colorectal cancer (CRC) face a grim prognosis. The goal of palliative intervention is directed at alleviating disease-related symptoms and improving quality of life. The provision of optimal palliative care for these patients is a compound and demanding process. This dilemma becomes more challenging when patients with advanced metastatic colorectal disease present with an incurable and asymptomatic primary lesion. Treatment options are numerous and include a variety of surgical and nonsurgical interventions. Most data regarding the role of surgery in palliation of CRC are from retrospective, nonrandomized case series. Surgical resection may provide good palliation of symptoms and prevent future tumor-related complications. Metal stents are also able to provide good palliative relief of obstruction and should be used when appropriate. The best palliative care will often require a multidisciplinary approach that involves input from surgical and nonsurgical teams, where treatment plans will be made in accordance with the wishes of the patient and family with a goal of decreasing morbidity and a focus on quality of life. PMID- 17913493 TI - Elucidating transient macromolecular interactions using paramagnetic relaxation enhancement. AB - Recent advances in the use of paramagnetic relaxation enhancement (PRE) in structure refinement and in the analysis of transient dynamic processes involved in macromolecular complex formation are presented. In the slow exchange regime, we show, using the SRY/DNA complex as an example, that the PRE provides a powerful tool that can lead to significant increases in the reliability and accuracy of NMR structure determinations. Refinement necessitates the use of an ensemble representation of the paramagnetic center and a model-free extension of the Solomon-Bloembergen equations. In the fast exchange regime, the PRE provides insight into dynamic processes and the existence of transient, low population intermediate species. The PRE allows one to characterize dynamic nonspecific binding of a protein to DNA; to directly demonstrate that the search process whereby a transcription factor locates its specific DNA target site involves both intramolecular (sliding) and intermolecular (hopping and intersegment transfer) translocation; and to detect and visualize the distribution of an ensemble of transient encounter complexes in protein-protein association. PMID- 17913496 TI - Immunoreceptor-like signaling by beta 2 and beta 3 integrins. AB - Although adhesion to extracellular structures is one of the most fundamental cell biological processes, the intracellular signals triggered by integrins, the most important receptors involved, are incompletely understood. Several recent reports indicate that signaling by beta(2) and beta(3) integrins in various cell types (neutrophils, macrophages, osteoclasts and platelets) use components of the signal transduction machinery of lymphocyte antigen receptors. Central to this immunoreceptor-like signaling is the phosphorylation of immunoreceptor tyrosine based activation motif (ITAM)-containing adapters (such as DAP12 and the Fc receptor gamma-chain) by Src-family kinases and the concomitant recruitment of the Syk tyrosine kinase through its dual SH2 domains. These and other reports reveal an unexpected similarity between the signal-transduction mechanisms used by integrins and immune recognition receptors. PMID- 17913497 TI - Peroxisomal dynamics. AB - Peroxisomes are a dynamic compartment in almost all eukaryotic cells and have diverse metabolic roles in response to environmental changes and cellular demands. The accompanying changes in enzyme content or abundance of peroxisomes are accomplished by dynamically operating membrane- and matrix-protein transport machineries. This review discusses recent progress in understanding peroxisomal proliferation and maintenance, insertion of peroxisomal membrane proteins, compartmentalization of peroxisomal matrix proteins and selective degradation of peroxisomes via pexophagy. PMID- 17913498 TI - The impact of offering a relatives' clinic on the satisfaction of the next-of-kin of critical care patients-a prospective time-interrupted trial. AB - BACKGROUND: Families have been shown to be dissatisfied with the frequency of doctor communication in the Critical Care setting. Discussions with families are often conducted in an ad-hoc fashion. We hypothesised that the offer of a formal daily weekday clinic for the relatives of Critical Care patients would increase the relatives' satisfaction. METHODS: All patients admitted to a 12-bedded Critical Care Unit over a 6-month period in a 480-bed hospital in the East of England were included in the study. The design was a time-interrupted prospective trial assessing the impact of an offered relatives' clinic on the satisfaction of the next-of-kin. Satisfaction was assessed 4 weeks following discharge from the Critical Care Unit by mail survey using the validated Critical Care Family Satisfaction Survey (1-5 scoring scale). RESULTS: Three hundred twenty-one patients met inclusion criteria. Survey return rate was 46%. The control and intervention groups were similar in size and demographics. Mean satisfaction of the control group was 4.50, and that of the intervention group was 4.55 (p=0.35). CONCLUSION: Offering a relatives' clinic does not significantly improve the satisfaction of the next-of-kin in this setting. Other means of improving satisfaction have been highlighted as a result of the study. PMID- 17913499 TI - The symmetry angle: a novel, robust method of quantifying asymmetry. AB - Quantification of asymmetry is a common objective in both research and clinical settings. The most common method for quantification of asymmetry of discrete variables is calculation of the symmetry index. Essentially a measure of the percent difference between sides, the symmetry index requires the choice of a reference value. This is a limitation as the choice of value is not always clear, and can lead to inconsistent results and artificially inflated values. Therefore, the purposes of the current study were to examine the limitations of the symmetry index in depth, define a new method of quantifying symmetry that is robust to those limitations (the symmetry angle), and compute the correlations between the two measures. The results showed that, when using the symmetry index, the interpretation of asymmetry can be highly affected by the choice of reference value. The symmetry angle does not require the choice of a reference value. Therefore, it is not prone to the same limitations. While symmetry angle values tend to be smaller than symmetry index values, the measures are very highly correlated. This suggests that the symmetry angle is a good substitute for the symmetry index. Future studies of asymmetry may benefit from the use of the symmetry angle, as it is equally effective for identifying intra-limb differences as the symmetry index, but is not prone to problems due to normalization and provides a standard scale (+/-100%) to interpret results. PMID- 17913500 TI - Reliability and validity of the Visual Gait Assessment Scale for children with hemiplegic cerebral palsy when used by experienced and inexperienced observers. AB - This study investigated the reliability and validity of the Visual Gait Assessment Scale when used by experienced and inexperienced observers. Four experienced and six inexperienced observers viewed videotaped footage of four children with hemiplegic cerebral palsy on two separate occasions. Validity of the Scale was obtained by comparison with three-dimensional gait analysis (3DGA). The experienced observers generally had higher inter-observer and intra-observer reliability than the inexperienced observers. Both groups showed higher agreement for assessments made at the ankle and foot than at the knee and hip. The experienced observers had slightly higher agreement with 3DGA than the inexperienced observers. The inexperienced observers showed a learning effect and had higher inter-observer agreement and higher agreement with 3DGA in the second assessment of the videotapes. This scale can be used by inexperienced observers but is limited to observations in the sagittal plane and by poor reliability at the knee and hip for experienced and inexperienced observers. PMID- 17913501 TI - Biological investigation of 131I-labeled new water soluble Ru(II) polypyridyl complex. AB - New [Ru(L1)(dcbpy)(NCS)2] complex was synthesized in a one-pot reaction starting from [RuCl2(p-cymene)]2, where the ligands (dcbpy=4,4'-dicarboxy-2,2'-bipyridine, L1=dipyrido[3,2-a:2',3'-c]phenazine-11-ylcarbonyl)-sodium) are introduced sequentially. The resulting complex was characterized by IR, NMR, and elemental analysis. The complex was labeled with I-131. Biodistribution study of the complex was carried out using 131I-labeled [Ru(L1)(dcbpy)(NCS)2] complex. The biodistribution study performed with albino Wistar male rats has shown that the complex has high uptake in the lung, small intestine, fat, and spleen. PMID- 17913502 TI - Determination of biological vector characteristics and nanoparticle dimensions for radioimmunotherapy with radioactive nanoparticles. AB - Radioimmunotherapy with biological vector labeled with radioactive nanoparticles is investigated from a dosimetric point of view. Beta (32P, 90Y) and low-energy X ray radionuclides (103Pd) are considered. Dose distributions inside solid tumors have been calculated using MCNPX 2.5.0. Nanoparticle dimensions and biological vector characteristics are also determined in order to reach the 50 Gy prescribed dose inside the entire tumor volume. The worst case of an avascular tumor is considered. Results show that for beta-emitting nanoparticles, a set of data (covering fraction, biological half-life, and nanoparticle radius) can be found within acceptable ranges (those of classical radioimmunotherapy). These sources (with Emax approximately few MeV) can be used for the treatment of tumors with a maximum diameter of about 1 cm. Low-energy X-rays (E<25 keV) can be used to extend the range of tumor diameter to 4-5 cm but require very tight biological vector characteristics. PMID- 17913503 TI - Significant improvement of cardiac sympathetic function following cardiac support device implantation: illustration by 123I-MIBG scintigraphy. AB - Cardiac support device implantation is a new and promising therapy for refractory heart failure. If functional and echocardiographic modifications have largely been reported following this procedure, there is a paucity of information in the clinical setting about cellular abnormalities modifications during the 'reverse remodeling' process. We report for the first time a clinical measurement of cardiac sympathetic function using 123I-MIBG scintigraphy following cardiac support device implantation. PMID- 17913504 TI - Thoracoscopic resection of bulky intrathoracic benign lesions. AB - BACKGROUND: Video-assisted thoracic surgery (VATS) is used for the diagnosis and treatment of some mediastinal lesions. However, large-size tumours are usually approached by thoracotomy or sternotomy. We report our experience of a full thoracoscopic approach for bulky intrathoracic lesions. METHODS: From November 2002 to March 2007, 14 patients with a bulky intrathoracic mass were referred for resection. The study group consisted of eight females and six males with a mean age of 44 years (range: 13-74). We defined as bulky a mass with a minimal cross sectional diameter equal to or larger than 50 mm, as measured on the specimen by the pathologist. RESULTS: Thoracoscopic resection was completed in all patients. In 4 cases, the mass originated from the pleura, and in 10 cases from the mediastinum. The larger diameter of the lesion ranged from 50 mm to 160 mm, with a median of 90.2 mm. Operative time, calculated from insertion of the first trocar to skin closure, ranged from 40 to 190 min (mean: 102). Mean chest drain duration was 2.1 days (range: 1-4 days) and the mean hospital stay was 4.3 days (range: 3-11 days). There were no major postoperative complications. The final pathological diagnoses were the following: solitary fibrous tumours of the pleura (4), benign thymic cysts (2), teratomas (2), bronchogenic cyst (1), benign thymoma (1), pleuropericardial cyst (1) and benign neurogenic tumours (3). CONCLUSIONS: With experience and use of appropriate instrumentation, resection of bulky intrathoracic lesions by thoracoscopy is feasible and safe. It should be considered as a reliable alternate for tumours that are benign and most often asymptomatic. PMID- 17913505 TI - Thoracoscopic localization techniques for patients with solitary pulmonary nodule: hookwire versus radio-guided surgery. AB - OBJECTIVE: Our aim was to evaluate the best intrathoracoscopic localization technique between hookwire and radio-guided surgery, in patients with pulmonary nodule. METHODS: From January 2000 to January 2005 we enrolled in this study 50 patients with a solitary pulmonary nodule, prospective randomized in two groups, well matched for diameter and depth of the pulmonary lesion. In 25 patients we performed the hookwire technique (Group A), whereas in the other 25 patients radio-guided localization was adopted (Group B). In both groups the localization technique was compared with finger palpation. In Group A, 9 lesions were in the left and 16 in the right lung; in Group B, 14 nodules were in the left lung and 11 in the right one. In both groups, the distance of the nodule from the pleural surface with lung inflated was 2.5 cm (1.5-2.5 cm in 12 patients, and >2.5 cm for the remaining 13). The mean size of the nodules in both groups was 1.1, range 0.6 1.9 (1 cm n=7 patients). RESULTS: All patients underwent thoracoscopic wedge resection, and 23 patients with a primary pulmonary lesion underwent thoracotomy for lobectomy and radical mediastinal lymphadenectomy. In Group A the hookwire technique localized the nodule in 21 of 25 patients (84%) whereas finger palpation localized it in 7 of 25 patients (28%). In Group B, radio-guided surgery localized the nodule in 24 of 25 patients (96%) whereas finger palpation localized it in 6 of 25 (24%). In Group A we registered 6 cases of pneumothorax compared to 1 case observed in the radio guided group. Postoperative hospital stay required an average of 4 days in both groups. CONCLUSIONS: In our experience radio-guided surgery has therefore been proven efficacious in the diagnosis of solitary pulmonary nodule and video assisted thoracoscopic surgery allows the removal of pulmonary nodules without complications. Hookwire was also shown to be efficacious but demonstrated complications linked primarily to external technical factors. PMID- 17913506 TI - Long-term experiences on cardiac retransplantation in adults. AB - BACKGROUND: It remains disputed whether cardiac retransplantation should be performed. This study aimed to evaluate our long-term experiences on cardiac retransplantation in adults. PATIENTS AND METHODS: Between March 1989 and December 2004, 2% (28/1290) of cardiac retransplantations were performed. RESULTS: The reasons for cardiac retransplantation were cardiac allograft vasculopathy (n=13; 47%), primary graft failure (n=11; 39%), and refractory acute rejection (n=4; 14%). The 30-day mortality risk was 29% (acute rejection: 50%; primary graft failure: 36%; cardiac allograft vasculopathy: 15%, p=0.324), compared to 8.5% for primary cardiac transplantation (p<0.001). The causes of early death were acute rejection (n=3; 37%), multiorgan failure (n=3; 37%), primary graft failure (n=1; 13%), and right ventricular failure (n=1; 13%). The late mortality rate was 96/1000 patient-years. The causes of late death were acute rejection (n=4; 50%), cardiac allograft vasculopathy (n=2; 25%), multiorgan failure (n=1; 13%), and infection (n=1; 13%). The 1-, 5-, 10-, and 15-year survival was respectively 78, 68, 54, and 38% (primary cardiac transplantation), and 46, 41, 32, and 32% (cardiac retransplantation) (p=0.003). The short-term survival for cardiac retransplantation due to cardiac allograft vasculopathy was likely better than primary graft failure and refractory acute rejection (p=0.09). CONCLUSION: The overall outcomes of cardiac retransplantation are significantly inferior to primary cardiac transplantation. Cardiac retransplantation should be only performed for selected patients. PMID- 17913507 TI - Visualization of entry and re-entry tears in a complex type A aortic dissection by 64-slice dual-source computer tomography. PMID- 17913509 TI - Basic interrupted versus continuous suturing techniques in bronchial anastomosis following sleeve lobectomy in dogs. AB - OBJECTIVE: Sleeve resection with or without lung resection is a valid conservative operation for patients with benign or malignant tumors; it enables the preservation of lung parenchyma. The aim of this prospective randomized study was to compare complications, operating time, and bronchial healing between the techniques of interrupted and continuous suturing for bronchial anastomosis in dogs. METHODS: Twenty adult mongrel dogs each weighing 18-22 kg (average: 20 kg) were divided into two groups according to the anastomosis technique performed: group A, interrupted suturing and group B, continuous suturing. Each group comprised of 10 dogs. Following right thoracotomy, sleeve resection of the right cranial lobe was performed in all dogs. Basic interrupted sutures using 4/0 Vicryl (Ethicon, USA) were used in group A, and continuous sutures were used in group B. RESULTS: The median anastomosis time was 15.2 min (range: 13-21 min) in group A and 9.6 min (range: 8-13 min) in group B. In all dogs, the anastomosis line was resected via right pneumonectomy for histopathological investigation 1 month after sleeve resection. Histopathological examination revealed that the healing of the anastomosis was not affected by the suturing technique applied. One dog from each group died on the fourth postoperative day; Fisher's exact test, p=0.763. CONCLUSIONS: Our research revealed that the healing of the anastomosis was not affected by the suturing technique performed. PMID- 17913510 TI - The inflammatory micro-environment in tumor progression: the role of tumor associated macrophages. AB - The link between inflammation and cancer proposed more than a century ago by Rudolf Virchow, who noticed the infiltration of leukocytes in malignant tissues, has recently found a number of genetic and molecular confirmations. Experimental, clinical and epidemiological studies have revealed that chronic inflammation contributes to cancer progression and even predisposes to different types of cancer. Cancer-associated inflammation includes: the presence of leukocyte infiltration; the expression of cytokines such as tumor necrosis factor (TNF) or interleukin (IL)-1; chemokines such as CCL2 and CXCL8; active tissue remodelling and neo-angiogenesis. Tumor-associated macrophages (TAM) are key regulators of the link between inflammation and cancer. Many observations indicate that, in the tumor micro-environment, TAM have several protumoral functions, including expression of growth factors, matrix proteases, promotion of angiogenesis and suppression of adaptive immunity. In this review we will discuss the role of TAM in the inflammatory micro-environment of solid tumors and will try to identify potential target for future therapeutic approaches. PMID- 17913511 TI - Benzoylnitrene radical anion: a new reagent for the generation of M-2H anions. AB - Benzoylnitrene radical anion, formed in high abundance by electron ionization of benzoylazide, is found to be a useful reagent for the formation of ionized reactive intermediates, such as diradicals and carbenes. The reactivity of the ion is similar to what is observed with atomic oxygen anion, occurring in many instances by H(2)(+) transfer. However, because benzoylnitrene radical anion is less basic than oxygen anion, it undergoes H(2)(+) transfer with substrates that react with oxygen anion by only proton transfer and therefore can be used to generate reactive ions not easily prepared by other methodologies. PMID- 17913512 TI - Differentiation of 1-O-alk-1'-enyl-2-acyl and 1-O-alkyl-2-acyl glycerophospholipids by multiple-stage linear ion-trap mass spectrometry with electrospray ionization. AB - We described linear ion-trap mass spectrometric approaches applying MS(3) and MS(4) toward to the structural characterization of 1-O-alk-1'-enyl-2-acyl-, 1-O alkyl-2-acyl-, and diacyl-glycerophospholipids (GPL) as the [M - H](-) ions desorbed by ESI in negative-ion mode. Further dissociation of the [lM - H - R(2)CO(2)H - polar head group](-) ions from the [M - H](-) ions of GPL that have undergone the consecutive losses of the fatty acid substituent at sn-2 and the polar head group readily gives the structural information of the radyl group at sn-1, resulting in structural differentiation among the 1-O-alk-1'-enyl-2-acyl-, 1-O-alkyl-2-acyl, and diacyl-glycerolphospholipid molecules. The distinction between a 1-O-alk-1'-enyl-2-acyl- and a 1-O-alkyl-2-acyl-GPL is based on the findings that the MS(3) (or MS(4)) spectrum of the [M - H - R(2)CO(2)H - polar head group](-) ion from the former compound is dominated by the alkenoxide anion that represents the radyl moiety at sn-1, while the spectrum from the latter compound is dominated by the ion at m/z 135 arising from further loss of the 1-O alkyl group as an alcohol. Another important notion is that the optimal collision energy required for acquiring the former spectrum is significantly lower than that required for obtaining the latter spectrum. Using the approaches, we are able to reveal the structures of several isobaric isomers in GPL mixtures of biological origin. Because the [M - H](-) ions are readily formed by various GPL classes (except glycerophosphocholine) in the negative-ion mode, these mass spectrometric approaches should have broad application in the structural identification of GPLs. PMID- 17913513 TI - Global familiarity of visual stimuli affects repetition-related neural plasticity but not repetition priming. AB - In this study, we tested the prediction of the component process model of priming [Henson, R.N. (2003). Neuroimaging studies of priming. Prog Neurobiol, 70 (1), 53 81] that repetition priming of familiar and unfamiliar objects produces qualitatively different neural repetition effects. In an fMRI study, subjects viewed four repetitions of familiar objects and globally unfamiliar objects with familiar components. Reliable behavioral priming occurred for both item types across the four presentations and was of a similar magnitude for both stimulus types. The imaging data were analyzed using multivariate linear modeling, which permits explicit testing of the hypothesis that the repetition effects for familiar and unfamiliar objects are qualitatively different (i.e., non-scaled versions of one another). The results showed the presence of two qualitatively different latent spatial patterns of repetition effects from presentation 1 to presentation 4 for familiar and unfamiliar objects, indicating that familiarity with an object's global structural, semantic, or lexical features is an important factor in priming-related neural plasticity. The first latent spatial pattern strongly weighted regions with a similar repetition effect for both item types. The second pattern strongly weighted regions contributing a repetition suppression effect for the familiar objects and repetition enhancement for the unfamiliar objects, particularly the posterior insula, superior temporal gyrus, precentral gyrus, and cingulate cortex. This differential repetition effect might reflect the formation of novel memory representations for the unfamiliar items, which already exist for the familiar objects, consistent with the component process model of priming. PMID- 17913514 TI - Magnetic resonance elastography of the brain. AB - The purpose of this study was to obtain normative data using magnetic resonance elastography (MRE) (a) to obtain estimates of the shear modulus of human cerebral tissue in vivo and (b) to assess a possible age dependence of the shear modulus of cerebral tissue in healthy adult volunteers. MR elastography studies were performed on tissue-simulating gelatin phantoms and 25 healthy adult volunteers. The data were analyzed using spatiotemporal filters and a local frequency estimating algorithm. Statistical analysis was performed using a paired t-test. The mean shear stiffness of cerebral white matter was 13.6 kPa (95% CI 12.3 to 14.8 kPa); while that of gray matter was lower at 5.22 kPa (95% CI 4.76 to 5.66 kPa). The difference was statistically significant (p<0.0001). PMID- 17913515 TI - Imaging brain and immune association accompanying cognitive appraisal of an acute stressor. AB - Acute stress elicits multiple responses in autonomic, endocrine, and immune systems. Cognitive appraisal is believed to be one important modulator of such stress responses. To investigate brain substrates of crosstalks between the homeostasis-maintaining systems accompanying appraisal of stressor controllability, we simultaneously recorded regional cerebral blood flow (rCBF) using 15O-water positron emission tomography, cardiovascular indices (heart rate (HR) and blood pressure (BP)), neuroendocrine indices (concentrations of epinephrine, norepinephrine, and adrenocorticotropic hormone (ACTH) in blood), and immune indices (proportions of subsets of lymphocytes (NK cells, helper T cells, cytotoxic T cells, and B cells) in blood), in 11 male subjects who performed a mental arithmetic task with either high controllability (HC) and low controllability (LC). The LC task resulted in less sense of control in subjects than the HC task. Significant increases of rCBF in the medial and lateral orbitofrontal cortices (OFC), and in the medial and lateral prefrontal cortices (MPFC, LPFC) were observed by subtracting the HC task from the LC task. More importantly, significant positive correlations between rCBF and HR, BP, and NK cells were commonly found in the OFC and MPFC during the LC tasks, but not during the HC tasks. The present results showed for the first time that the prefrontal neural network including the OFC and MPFC might be one pivotal region for bi directional functional association between the brain and peripheral autonomic and immune activities accompanying appraisal of an acute stressor. PMID- 17913517 TI - The impact of a citywide audit with educational intervention on the care of patients with epilepsy. AB - The care of patients with epilepsy historically has been well documented to be poor. Previous attempts to improve care through education have been unsuccessful. The New GP Contract in the UK introduced epilepsy as a core quality indicator from April 2004. This prospective audit assesses the impact of an audit with educational intervention on the process of care of patients with epilepsy. The case notes of 610 patients, of all ages, with epilepsy on treatment, in 13 general practices serving Chester and surrounding area were reviewed before and 2 years after an intervention, comprising (a) the provision of a comprehensive template, (b) individualised categorisation for each patient and (c) an educational session led by a Neurologist. The overall review rate increased in the first year from 41 to 49% (p<0.0001) and by 2 years to 63% (p<0.0001). Documented remission rate increased from 29 to 43% (p<0.0001). Admissions to accident and emergency fell significantly (p=0.0026). There was no fall in the non-compliance rate. Forty five percent of patients with documented poor control were not under shared care. Issues highlighted in the audit generated 77 referrals. There were clear health gains in 62 (13%) individuals from referrals and practice interventions related to the audit. This original audit identified significant improvements in review rate, documented remission rate and beneficial outcomes in individual patients. The changes were attributable to both the educational intervention and the coincidental acceptance of the New GP Contract. Remaining problems include lack of shared care for patients with active epilepsy. PMID- 17913516 TI - Seizure-freedom with combination therapy in localization-related epilepsy. AB - We analyzed the effect of combination therapy on seizure frequency in all adult patients (N=193) with focal epilepsy followed at a single institution in a cross sectional study. One hundred and thirty-five patients were on two AEDs, of them, 37 (27%) were seizure-free, 50 patients were on three AEDs including 5 (10%) seizure-free patients (p<0.01 for seizure-freedom with two AEDs versus three AEDs). Thirty-five different combinations were used in patients on two AEDs and 40 combinations on patients on three drugs emphasizing the difficulties involved in evaluation of the efficacy and tolerability of specific combinations. The significant proportion of seizure-free cases (27%) on duotherapy is suggesting the usefulness of combination therapy in achieving seizure-freedom in epilepsies refractory to single drug treatment. The material in the study was not from a randomized trial and therefore the comparability of patients on different AEDs is uncertain, but on the other hand the clinical practice followed provides a natural experiment suitable for comparative, non-randomized assessment of treatment outcomes. PMID- 17913518 TI - Detection of venous emboli using Doppler ultrasound. AB - OBJECTIVES: To detect emboli in the venous system using a Doppler ultrasound device with embolism detecting software. METHODS: Pulmonary embolism (PE) was induced by injecting thrombus through the iliac vein of castrated swine under general anaesthesia. Data recorded from the Doppler system were analysed for high intensity transient signals (HITS) using receiver operating characteristic curves. Four different thrombi (5 or 10mm long and 3 or 5mm in diameter) were then injected to assess the quantitative analysis. RESULTS: Thrombus could be detected in the venous system by the Doppler ultrasound device with an embolism detecting function. Appropriate confidence level was 60%. If thrombus were assumed to travel at the maximum flow rate (30 cm/s) in the inferior vena cava, the estimated embolism size was 10.4 S.D. 2.8mm for 3mm and 10.8 S.D. 4.9 mm for 5mm, both of which were close to 10mm. CONCLUSION: Thrombi could be detected as high intensity transient signals in the venous system. The appropriate confidence level was 60%. The size of emboli can be estimated if they are more than 3mm in diameter when the venous flow rate is 30 cm/s or less. Our results suggest that it may be possible to detect emboli in the subclavian vein, axillary vein or inferior vena cava in clinical cases. PMID- 17913519 TI - Endovascular stent-graft repair for spontaneous rupture of the nonaneurysmal thoracic aorta in a patient with Takayasu's arteritis. AB - Takayasu's arteritis is a disease of unknown etiology with a constellation of clinical findings primarily resulting from stenotic lesions on the aorta and its branches. Although aneurysmal degeneration is observed frequently in patients with Takayasu's arteritis, non-aneurysmal spontaneous aortic rupture is extremely rare. We report a case of endovascular stent grafting for spontaneous rupture of a non-aneurysmal thoracic aorta in Takayasu's arteritis. PMID- 17913520 TI - Five day antibiotic prophylaxis for major lower limb amputation reduces wound infection rates and the length of in-hospital stay. AB - OBJECTIVE: To compare wound infection, revision rates and hospital stay after major lower limb amputation between patients receiving 24 hours versus 5 days of prophylactic antibiotics. METHODS: The outcomes of a consecutive series of 40 major lower limb amputations in patients receiving a short 24-hour course of combined prophylactic antibiotics (flucloxacillin/vancomycin + gentamicin/ciproxin + metronidazole) were retrospectively analysed. Following this a further consecutive group of 40 major lower limb amputations were studied prospectively following the institution of a 5-day combined regime using the same antibiotics. RESULTS: The 2 groups of patients were similar in terms of demographics, vascular risk factors and level of amputation. The 5-day antibiotic regime led to a significant reduction in wound infection rates (5% vs. 22.5%, P=0.023) and a reduced length of hospital stay (22 vs. 34 days, P=0.001). Revision rates were lower (2.5% vs. 10%) but did not reach statistical significance (P=0.36). More patients in the prospective 5-day antibiotic series were operated on by the vascular trainee. (77.5% vs. 55% P=0.033). CONCLUSIONS: This data supports the use of a prolonged 5-day course of combined antibiotics after major lower limb amputation. This appears to reduce stump infection rates leading to shorter in-hospital stay. PMID- 17913522 TI - Inferior mesenteric artery aneurysm with occlusion of the superior mesenteric artery, coeliac trunk and right renal artery. AB - Inferior mesenteric artery aneurysms are amongst the rarest of visceral aneurysms. We present here a case associated with occlusion of the superior mesenteric artery, coeliac trunk and right renal artery. Operative treatment was resection of the aneurysm, with end-to-end anastomosis. This is the first description of this condition from the UK, with only nine other reports worldwide. Such pathology may be caused by a "jet disorder" phenomenon, with increased flow through the inferior mesenteric artery due to chronic mesenteric occlusive disease. PMID- 17913523 TI - Significant increase of self-renewal in hematopoietic cells after forced expression of EVI1. AB - EVI1 was first identified as a preferential integration site of ecotropic retroviruses in the MDS1/EVI1 genomic locus leading to myeloid tumors in susceptible mice. Later studies showed that retroviral integration in the MDS1/EVI1 locus results in the emergence of a non-malignant dominant hematopoietic stem cell clone in non-susceptible mice strains, in non-human primates, and in patients, suggesting that a gene encoded by the locus could affect the self-renewal potential of a cell. The locus encodes two genes. One of them, EVI1, has long been associated with myeloid leukemia. To understand whether EVI1 has a role in self-renewal control, we forcibly expressed EVI1 in the bone marrow progenitors of two mice strains that differ in their proliferation and self-renewal potential. By comparing the response of the hematopoietic cells to EVI1, we conclude that EVI1 has a role in prolonging the self-renewal potential of the cells and that this ability of EVI1 is limited and modulated by inherent characteristics of the cells. PMID- 17913524 TI - Subfoveal choroidal neovascularization in a 3-year-old child with North Carolina macular dystrophy. AB - North Carolina macular dystrophy is characterized by nonprogressive atrophy of the choroid, choriocapillaris, and the retinal pigment epithelial layer. The characteristic retinal findings, ranging from scattered drusen to a posterior staphyloma, have been reported as early as infancy and usually reach their greatest magnitude in the second decade of life. Herein, we describe a case of a 3-year-old boy with a documented family history of North Carolina macular dystrophy who presented with a subfoveal choroidal neovascular membrane in addition to the macular dystrophic changes. PMID- 17913525 TI - Localization and visualization of excess chemical potential in statistical mechanical integral equation theory 3D-HNC-RISM. AB - In this study the excess chemical potential of the integral equation theory, 3D RISM-HNC [Q. Du, Q. Wei, J. Phys. Chem. B 107 (2003) 13463-13470], is visualized in three-dimensional form and localized at interaction sites of solute molecule. Taking the advantage of reference interaction site model (RISM), the calculation equations of chemical excess potential are reformulized according to the solute interaction sites s in molecular space. Consequently the solvation free energy is localized at every interaction site of solute molecule. For visualization of the 3D-RISM-HNC calculation results, the excess chemical potentials are described using radial and three-dimensional diagrams. It is found that the radial diagrams of the excess chemical potentials are more sensitive to the bridge functions than the radial diagrams of solvent site density distributions. The diagrams of average excess chemical potential provide useful information of solute-solvent electrostatic and van der Waals interactions. The local description of solvation free energy at active sites of solute in 3D-RISM-HNC may broaden the application scope of statistical mechanical integral equation theory in solution chemistry and life science. PMID- 17913526 TI - Inhibition of iNOS expression and NO production by anti-inflammatory steroids. Reversal by histone deacetylase inhibitors. AB - In inflammation, nitric oxide (NO) is produced by inducible nitric oxide synthase (iNOS) induced by bacterial products and cytokines, and NO acts as a regulatory and pro-inflammatory mediator. Glucocorticoids are powerful anti-inflammatory agents that inhibit the expression of iNOS and various other inflammatory factors. Histone deacetylation has been recently described as a novel mechanism how glucocorticoids down-regulate transcriptional activation of some inflammatory genes. The aim of the present study was to investigate the effects of inhibitors of histone deacetylation on the suppressive effects of glucocorticoids on NO production and iNOS expression. Dexamethasone and a dissociated glucocorticoid RU24858 inhibited NO production, and iNOS protein and mRNA expression in macrophages exposed to bacterial lipopolysaccharide (LPS). In the presence of a glucocorticoid receptor (GR) antagonist mifepristone, dexamethasone and RU24858 had no effect on NO production. The role of histone deacetylation in the glucocorticoid effect was studied by using three structurally different inhibitors of histone deacetylases (HDACs): trichostatin A, apicidin and MC1293. HDAC inhibitors reversed the effects of dexamethasone and RU24858 on iNOS expression and NO production. Stably transfected A549/8 cells containing luciferase gene under the control of human iNOS promoter were used in promoter activity studies. iNOS promoter activity induced by IL-1beta was inhibited by dexamethasone and the inhibitory effect was reversed by HDAC inhibitor trichostatin A. The results suggest that glucocorticoids inhibit iNOS expression and NO production by a GR-mediated and GRE-independent manner through histone deacetylation and transcriptional silencing. PMID- 17913527 TI - Effects of meal size, clutch, and metabolism on the energy efficiencies of juvenile Burmese pythons, Python molurus. AB - We explored meal size and clutch (i.e., genetic) effects on the relative proportion of ingested energy that is absorbed by the gut (apparent digestive efficiency), becomes available for metabolism and growth (apparent assimilation efficiency), and is used for growth (production efficiency) for juvenile Burmese pythons (Python molurus). Sibling pythons were fed rodent meals equaling 15%, 25%, and 35% of their body mass and individuals from five different clutches were fed rodent meals equaling 25% of their body mass. For each of 11-12 consecutive feeding trials, python body mass was recorded and feces and urate of each snake was collected, dried, and weighed. Energy contents of meals (mice and rats), feces, urate, and pythons were determined using bomb calorimetry. For siblings fed three different meal sizes, growth rate increased with larger meals, but there was no significant variation among the meal sizes for any of the calculated energy efficiencies. Among the three meal sizes, apparent digestive efficiency, apparent assimilation efficiency, and production efficiency averaged 91.0%, 84.7%, and 40.7%, respectively. In contrast, each of these energy efficiencies varied significantly among the five different clutches. Among these clutches production efficiency was negatively correlated with standard metabolic rate (SMR). Clutches containing individuals with low SMR were therefore able to allocate more of ingested energy into growth. PMID- 17913528 TI - Characterization and expression of Peroxiredoxin 1 in the neonatal tammar wallaby (Macropus eugenii). AB - Peroxiredoxin 1 (PRDX1) is a ubiquitously expressed antioxidant with vital roles in basal metabolic functions. In addition PRDX1 is involved in cell differentiation and proliferation, apoptosis and innate immunity. In this study, we have characterized PRDX1 from the tammar wallaby (Macropus eugenii). Tammar PRDX1 has high conservation of functional residues and motifs, and demonstrates a close homology with eutherian and vertebrate orthologues. Stimulation of adult tammar leukocytes with lipopolysaccharide and lipoteichoic acid suggests a role for PRDX1 in innate immune defences. PRDX1 expression in the organs of tammar pouch young was mildly elevated early in life possibly reflecting its role in basal metabolic processes. Later increases in PRDX1 expression correlated with functional maturation of several immune organs or with preparation for increased oxidative stress of emergence. The findings of the study are reflections of the complex integrated roles that PRDX1 has in regulation of oxidative stress, apoptosis, cell differentiation and proliferation, and innate immunity. PMID- 17913530 TI - Identification and tissue expression analysis of C-type lectin and galectin in the Pacific oyster, Crassostrea gigas. AB - As an initial step in the functional analysis of lectins in the Pacific oyster, Crassostrea gigas, we attempted to obtain the full coding sequences of C. gigas lectins and conduct tissue expression analyses. To obtain lectin genes quickly, we identified C. gigas expressed sequence tags that coded for lectins in GenBank, and selected three encoding partial sequences of C-type lectin 1 (CgCLec-1), galectin (CgGal) and fucolectin. We obtained full open reading frames of CgCLec-1 and CgGal cDNAs by RACE-PCR. CgCLec-1 is a typical C-type lectin with a signal peptide and C-type lectin domain. CgCLec-1 mRNA was expressed only in specialized basophilic cells involved with digestive enzyme secretion in the digestive gland, suggesting that CgCLec-1 is secreted into the lumen of the digestive diverticula. CgGal is a prototype galectin with a single galactose-binding domain that was expressed in all of the tissues examined. As suggested for vertebrate galectin-1 (prototype galectin), CgGal may function in general cell activities such as cell adhesion. Fucolectin in C. gigas was expressed specifically in the gonads, indicating a possible function in gonadal development. CgCLec-1 and CgGal expression in hemocytes was not upregulated after injecting Vibrio tubiashii into adductor muscle, suggesting that bacterial infection does not induce synthesis of these lectins. Of the three lectins examined, CgCLec-1 is an interesting target for future investigations of innate immunity in the digestive system of C. gigas. PMID- 17913531 TI - A review of feral cat control. AB - Animal overpopulation including feral cats is an important global problem. There are many stakeholders involved in the feral cat debate over 'what to do about the problem', including those who consider them a nuisance, the public at risk from zoonotic disease, people who are concerned about the welfare of feral cats, those concerned with wildlife impacts, and the cats themselves. How best to control this population is controversial and has ranged from culling, relocation, and more recently 'trap neuter return' (TNR) methods. Data support the success of TNR in reducing cat populations, but to have a large impact it will have to be adopted on a far greater scale than it is currently practised. Non-surgical contraception is a realistic future goal. Because the feral cat problem was created by humans, concerted educational efforts on responsible pet ownership and the intrinsic value of animals is an integral part of a solution. PMID- 17913532 TI - Use of latissimus dorsi and abdominal external oblique muscle for reconstruction of a thoracic wall defect in a cat with feline osteochondromatosis. AB - A 4-year-old, male castrated European shorthair cat was presented with a firm mass palpable on the right caudal rib cage. Lateral and ventrodorsal radiographs of the thorax revealed a 4x3x2cm large, expansile and radiodense mass originating from the distal part of the 13th rib. After removal of the tumour, which was histopathologically confirmed as feline osteochondromatosis, the diaphragm, omentum, external abdominal oblique and latissimus dorsi muscles were used to reconstruct the defect. Feline osteochondromatosis is induced by retroviruses, eg, feline leukaemia virus, for which the cat tested positive. The tumour was removed for palliative reasons, because such tumours have the tendency to transform into osteosarcomas. Six months after the surgical excision the cat showed no clinical signs of reoccurrence. PMID- 17913529 TI - Ethanol teratogenesis in Japanese medaka: effects at the cellular level. AB - The adverse effects of alcohol on the developing humans represent a spectrum of structural and neurobehavioral abnormalities, most appropriately termed as fetal alcohol spectrum disorder (FASD). The mechanism by which ethanol induces FASD is unknown. Human studies of FASD are very limited due to ethical constraints; however, several animal models from nematodes to mammals are utilized to understand the molecular mechanism of this disorder. We have used Japanese medaka (Oryzias latipes) embryo-larval development as a unique non-mammalian model to study the molecular mechanism of FASD. Fertilized medaka eggs were exposed to ethanol (0-400 mM) for 48 h post fertilization (hpf) and then maintained in regular embryo rearing medium without ethanol. Viable embryos were harvested on 0, 2, 4 and 6 day post fertilization (dpf) and analyzed for DNA, RNA and protein contents of the embryos. By applying semi-quantitative RT-PCR (rRT-PCR) and quantitative real-time RT-PCR (qRT-PCR), RNA samples were further analyzed for seven transcription factors, emx2, en2, iro3, otx2, shh, wnt1 and zic5 which are expressed in the neural tube of medaka embryo during early phase of development. RNA and protein contents of the embryos were significantly reduced by ethanol at 400 mM dose on 4 and 6 dpf compared to the control (no ethanol), and 100 mM ethanol treated embryos. However, significant reduction of DNA was observed only in 4 dpf embryos. Total protein contents of yolk remained unaltered after ethanol treatment. Expression pattern of emx2, en2, iro3, otx2, shh, wnt1, and zic5 mRNAs were found to be developmentally regulated, however, remained unaltered after ethanol treatment. It is therefore concluded that alteration of nucleic acid and protein contents of medaka embryo by ethanol could be used as an indicator of embryonic growth retardation which might be the result of disruption of specific gene function during development. PMID- 17913533 TI - Influenza viruses and the evolution of avian influenza virus H5N1. AB - Although small in size and simple in structure, influenza viruses are sophisticated organisms with highly mutagenic genomes and wide antigenic diversity. They are species-specific organisms. Mutation and reassortment have resulted in newer viruses such as H5N1, with new resistance against anti-viral medications, and this might lead to the emergence of a fully transmissible strain, as occurred in the 1957 and 1968 pandemics. Influenza viruses are no longer just a cause of self-limited upper respiratory tract infections; the H5N1 avian influenza virus can cause severe human infection with a mortality rate exceeding 50%. The case death rate of H5N1 avian influenza infection is 20 times higher than that of the 1918 infection (50% versus 2.5%), which killed 675000 people in the USA and almost 40 million people worldwide. While the clock is still ticking towards what seems to be inevitable pandemic influenza, on April 17, 2007 the U.S. Food and Drug Administration (FDA) approved the first vaccine against the avian influenza virus H5N1 for humans at high risk. However, more research is needed to develop a more effective and affordable vaccine that can be given at lower doses. PMID- 17913534 TI - Epstein-Barr virus infection with acute pancreatitis. PMID- 17913535 TI - Eight years of hepatitis B vaccination in Colombia with a recombinant vaccine: factors influencing hepatitis B virus infection and effectiveness. AB - OBJECTIVE: To evaluate the effectiveness of a recombinant hepatitis B vaccine used in endemic areas of Colombia, as well as risk factors associated with hepatitis B virus (HBV) infection and carriage after vaccine introduction. METHODS: A cross-sectional study was carried out in urban and rural areas of the Colombian Amazon, a highly endemic area for hepatitis B infection. Children under 12 years of age and their mothers were selected for the study using one-stage cluster sampling (N=2145) and were examined for HBV serological markers and antibodies against surface antigen (anti-HBs). RESULTS: There has been a reduction of 60-75% in the prevalence of HBV infection and hepatitis B surface antigen (HBsAg) carriage since HBV vaccination was introduced. Receiving the first dose of HBV vaccine at more than two months after birth was one of the factors associated with HBV carrier status. Maternal HBV infection was also associated with infection in the child. CONCLUSIONS: The recombinant Cuban hepatitis B vaccine has contributed to the reduction of the infection in this highly endemic area, though further efforts are required to improve timely vaccination for children at high risk. PMID- 17913536 TI - Presence and characterization of Mycobacterium avium subspecies paratuberculosis from clinical and suspected cases of Crohn's disease and in the healthy human population in India. AB - OBJECTIVES: To investigate and characterize Mycobacterium avium subspecies paratuberculosis (MAP) in patients with Crohn's disease, attendants of animals with suspected infection, and healthy humans, using multiple diagnostic tests. METHODS: A total of 119 samples (35 stool, 76 serum, three blood clots, and five biopsies) were collected from five patients with Crohn's disease, eight attendants of animals with Johne's disease, and 93 apparently normal control subjects (Agra region) from North India. Samples were screened for the presence of MAP by smear examination, culture of stool, blood clot and biopsies, and ELISA. Colonies obtained by culture were further characterized using polymerase chain reaction (PCR) with IS900 MAP-specific primers. RESULTS: Using all diagnostic modalities, MAP and/or MAP antibodies were identified in 100% (5/5) of subjects with Crohn's disease; 75.0% (6/8) of attendants of MAP infected animals were positive and 38.0% (27/71) of apparently normal controls were also positive. Most sensitive test was ELISA (100%, 5/5), followed by culture (80.0%, 4/5), and acid-fast staining. Ziehl-Neelsen staining was positive in 37.5% (3/8) of subjects with active animal husbandry practices. In 71 serum samples from control subjects, seroprevalence of MAP was 38.0% using indigenous protoplasmic antigens (PPA) and 36.6% using commercial PPA. Of the serum samples from the Crohn's disease patients, 100% (5/5) were positive by ELISA using indigenous PPA and 40.0% (2/5) were positive by ELISA using commercial PPA. IS900 PCR was used to characterize tiny colonies of MAP that grew extremely slowly on Herrold's egg yolk medium, and of 15 (42.8%) cultures, 14 (93.3%) were typed as MAP. CONCLUSIONS: Paper documented the presence of MAP in all patients with Crohn's disease, in some animal attendants who had the history of working with goat herds infected with Johne's disease and in few normal healthy individuals. Presence of Ziehl Neelsen positive MAP. In the stool of attendants working with MAP-infected animals was unique to humans. ELISA based on antigens derived from indigenous MAP 'bison type' genotype of goat origin was most sensitive modality for screening Crohn's disease patients. PMID- 17913537 TI - Antibody response against saliva antigens of Anopheles gambiae and Aedes aegypti in travellers in tropical Africa. AB - Exposure to vectors of infectious diseases has been associated with antibody responses against salivary antigens of arthropods among people living in endemic areas. This immune response has been proposed as a surrogate marker of exposure to vectors appropriate for evaluating the protective efficacy of antivectorial devices. The existence and potential use of such antibody responses in travellers transiently exposed to Plasmodium or arbovirus vectors in tropical areas has never been investigated. The IgM and IgG antibody responses of 88 French soldiers against the saliva of Anopheles gambiae and Aedes aegypti were evaluated before and after a 5-month journey in tropical Africa. Antibody responses against Anopheles and Aedes saliva increased significantly in 41% and 15% of the individuals, respectively, and appeared to be specific to the mosquito genus. A proteomic and immunoproteomic analysis of anopheles and Aedes saliva allowed for the identification of some antigens that were recognized by most of the exposed individuals. These results suggest that antibody responses to the saliva of mosquitoes could be considered as specific surrogate markers of exposure of travellers to mosquito vectors that transmit arthropod borne infections. PMID- 17913538 TI - Arginine-specific gingipain A from Porphyromonas gingivalis induces Weibel-Palade body exocytosis and enhanced activation of vascular endothelial cells through protease-activated receptors. AB - Gingipains, cysteine proteases derived from Porphyromonas gingivalis, are important virulence factors in periodontal diseases. We found that arginine specific gingipain A (RgpA) increased the responsiveness of vascular endothelial cells to P. gingivalis lipopolysaccharides (LPS) and P. gingivalis whole cells to induce enhanced IL-8 production through protease-activated receptors (PARs) and phospholipase C (PLC) gamma. We therefore investigated whether RgpA-induced enhanced cell activation is mediated through exocytosis of Weibel-Palade bodies (WPBs) because they store vasoactive substances. RgpA rapidly activated PAR- and PLCgamma-dependent WPB exocytosis. In addition, angiopoietin (Ang)-2, a substance of WPB, enhanced IL-8 production by P. gingivalis LPS, suggesting that Ang-2 mediates the RgpA-induced enhanced cell responses. Thus, we propose a novel role for RgpA in induction of a proinflammatory event through PAR-mediated WPB exocytosis, which may be an important step for enhanced endothelial responses to P. gingivalis. PMID- 17913539 TI - Chlamydophilal antigens induce foam cell formation via c-Jun NH2-terminal kinase. AB - Chlamydophila pneumoniae is known to be associated with atherosclerosis. Recent studies have reported that components of Chlamydophila pneumoniae (chlamydophilal antigens) induce foam cell formation in macrophages. However, the mechanism of foam cell formation induced by chlamydophilal antigens has yet to be elucidated. In this paper, we first found that mitogen-activated protein kinases including extracellular signal-regulated kinase, p38 and c-Jun NH2 terminal kinase are phosphorylated after stimulation by chlamydophilal antigens. We then showed that chlamydophilal antigens induce foam cell formation mainly via c-Jun NH2 terminal kinase. Finally, we demonstrated that foam cell formation and phosphorylation of mitogen-activated protein kinases induced by chlamydophilal antigens are mainly recognized through Toll-like receptor 2. These results collectively indicated that chlamydophilal antigens induce foam cell formation mainly via Toll-like receptor 2 and c-Jun NH2 terminal kinase. PMID- 17913540 TI - Extended immunization intervals enhance the immunogenicity and protective efficacy of plasmid DNA vaccines. AB - Effective vaccines against infectious diseases and biological warfare agents remain an urgent public health priority. Studies have characterized the differentiation of effector and memory T cells and identified a subset of T cells capable of conferring enhanced protective immunity against pathogen challenge. We hypothesized that the kinetics of T cell differentiation influences the immunogenicity and protective efficacy of plasmid DNA vaccines, and tested this hypothesis in the Plasmodium yoelii murine model of malaria. We found that increasing the interval between immunizations significantly enhanced the frequency and magnitude of CD8+ and CD4+ T cell responses as well as protective immunity against sporozoite challenge. Moreover, the interval between immunizations was more important than the total number of immunizations. Immunization interval had a significantly greater impact on T cell responses and protective immunity than on antibody responses. With prolonged immunization intervals, T cell responses induced by homologous DNA only regimens achieved levels similar to those induced by heterologous DNA prime/ virus boost immunization at standard intervals. Our studies establish that the dosing interval significantly impacts the immunogenicity and protective efficacy of plasmid DNA vaccines. PMID- 17913541 TI - Identification of proteins directly phosphorylated by UL13 protein kinase from herpes simplex virus 1. AB - Herpes simplex virus 1 (HSV-1) UL13 is a viral protein kinase that regulates optimal viral replication in cell cultures. Identification of substrates of protein kinases is a crucial step to elucidate the mechanism by which they function. Using our developed system to analyze the specific protein kinase activity of UL13, we have shown that UL13 protein kinase directly phosphorylates the viral proteins ICP22 and UL49 previously reported to be putative substrates. We also identified UL41 as a previously unreported and novel substrate of UL13. These data will serve as a basis to clarify the mechanism by which UL13 influences viral replication. PMID- 17913542 TI - CD44, a signal receptor for the inhibition of the cytoadhesion of CD36-binding Plasmodium falciparum-infected erythrocytes by CSA-binding infected erythrocytes. AB - The cytoadhesion of Plasmodium falciparum-infected erythrocytes (IEs) in organ microvessels is a key event in the pathogenesis of cerebral malaria and pulmonary edema. Identification of the molecules involved in the interaction between IEs and endothelial cells has been a major goal of research into severe forms of malaria. In contrast, the consequences of cytoadhesion for endothelial cells have been largely ignored. By combining phenotypic selection, cytoadhesion assays and flow cytometry, we demonstrated that the cytoadhesion of CSA-binding IEs inhibited the cytoadhesion of CD36-binding IEs. We identified CD44 as a signal receptor for CSA-binding IEs cytoadhesion, and demonstrated that the signal was transduced to CD36 through a pathway involving the Src-kinase family and MEK. CD36-mediated cytoadhesion was modulated independently of changes in CD36 expression. These results provide the first evidence that some IEs can downregulate the cytoadhesion of IEs of another phenotype, by modifying endothelial cells via a signaling pathway relating CD44 to CD36. Mimicking this phenomenon may constitute an interesting therapeutic strategy for inhibiting the adhesion of CD36-binding IEs -- the most abundant phenotype among field isolates - and promoting their degradation in the spleen. PMID- 17913544 TI - Pre-existing anti-Salmonella vector immunity prevents the development of protective antigen-specific CD8 T-cell frequencies against murine listeriosis. AB - Our laboratory has focused its research on the use of the type III secretion system of Salmonella enterica serovar Typhimurium to translocate heterologous antigens directly into the cytosol of antigen-presenting cells. We have previously reported that the single oral immunization of mice with a recombinant Salmonella aroA/sptP mutant strain expressing the translocated Yersinia outer protein E fused to the immunodominant antigen p60 from Listeria monocytogenes in a type III-mediated fashion results in the efficient induction of p60-specific CD8 T cells and confers protection against a lethal Listeria challenge infection. In the present study, we determined whether pre-existing anti-Salmonella vector immunity influences the induction of p60-specific CD8 T cells and modulates protective immunity against listeriosis after oral vaccination with recombinant Salmonella. After single oral immunization, the Salmonella aroA/sptP double mutant strain was found to colonize spleens of mice for 21days. In contrast, the period of colonization was significantly shortened to 6days due to anti Salmonella vector immunity after second oral immunization. The latter scenario led to the induction of low-level frequencies of antigen-specific CD8 T cells. Compared to the significantly higher numbers of p60-specific T lymphocytes elicited after single oral immunization, the low amount of Listeria-specific CD8 T cells did not confer protection against listeriosis. PMID- 17913543 TI - A surface 75-kDa protein with acid phosphatase activity recognized by monoclonal antibodies that inhibit Paracoccidioides brasiliensis growth. AB - Paracoccidioides brasiliensis is a thermo-dimorphic fungus responsible for paracoccidioidomycosis (PCM), a systemic granulomatous mycosis prevalent in Latin America. The fungus releases many antigens which may be transiently bound to its cell surface. Some of them may show enzymatic functions essential for maintaining many cell processes and survival of the microorganism at different conditions. In this study, we report the characterization of a secreted 75kDa protein from P. brasiliensis with phosphatase activity. Biologic function of the molecule was demonstrated using two specific mAbs produced and characterized as IgM and IgG isotypes. Confocal microscopy and flow cytometry analysis demonstrated that both mAbs recognized the protein on the fungus surface, mainly in its budding sites. In vitro experiments showed that fungal growth was inhibited by blocking the protein with mAbs. In addition, opsonized yeast cells with both mAbs facilitated phagocytosis by murine peritoneal macrophages. Passive immunization using mAbs before P. brasiliensis mice infection reduced colony-forming units (CFU) in the lungs as compared with controls. Histopathology showed smaller inflammation, absence of yeast cells and no granuloma formation. PMID- 17913546 TI - The role of human innate immune factors in nasal colonization by Staphylococcus aureus. AB - Staphylococcus aureus colonization of the human nares predisposes to sometimes severe auto-infection. To investigate whether genetic polymorphism affects the S. aureus carriage status, sequence variation in alpha-defensin and beta-defensin, and mannose-binding lectin (MBL) genes were determined for a group of volunteers (n=109) with known S. aureus nasal carriage status. DEFA1/3 expression was measured in a subset of the volunteers (n=32). None of the single nucleotide polymorphisms studied could clearly distinguish the (non) carriage groups. S. aureus carriers differed from non-carriers in baseline level of HNP1-3 peptide production (median: 218 versus 89mug/ml, P=0.016). No association between HNP1-3 levels and the individual sequence polymorphisms was documented. The combined copy numbers of DEFA1/A3 genes ranged from 5 to 23 per diploid genome. A linear correlation between combined copy numbers and HNP1-3 peptide concentrations in nasal secretions of non-carriers was noted (r(2)=0.8991). DEFA3 gene was absent in 25% of the individuals. MBL haplotype A was overrepresented in persistent S. aureus carriers (87% vs. 67%; P=0.038). In conclusion, defensin gene polymorphism, both in sequence and in gene copy numbers, does not seem to be involved in S. aureus carriage predisposition. However, MBL haplotypes do so significantly. Baseline HNP1-3 production is more the consequence of S. aureus colonization than a reason for the (non) carrier status. PMID- 17913545 TI - Systemic but not mucosal immunity induced by AVA prevents inhalational anthrax. AB - Improved vaccines and adjuvants are being developed to reduce the threat posed by a terrorist attack involving aerosolized anthrax spores. Nevertheless, uncertainty persists concerning the relative benefits of inducing mucosal vs systemic immunity to host survival following inhalational exposure to anthrax spores. This work examines the effect of delivering the licensed human vaccine (anthrax vaccine adsorbed, AVA) combined with a CpG oligodeoxynucleotide (ODN) adjuvant intraperitoneally or intranasally to A/J mice. Results indicate that protection from inhalational anthrax correlates with the induction of a strong systemic rather than mucosal immune response, and demonstrate that protection is significantly improved and accelerated by the addition of CpG ODN. PMID- 17913548 TI - The bacterial metabolite 2,3-butanediol ameliorates endotoxin-induced acute lung injury in rats. AB - Widely identified in bacteria, yeasts and human beings, 2,3-butanediol has been studied for decades. This chemical reportedly functions as a neutralization agent to counteract lethal acidification by bacterial growth and as a signaling molecule involved in interactions among insects, and between bacteria and the plant host. While 2,3-butanediol is produced by many pathogenic bacterial species, its significance and effect on mammals remains basically uncharacterized. Herein, we show that gastric intubation of 2,3-butanediol in rats significantly ameliorates acute lung injury (ALI) and the inflammatory responses induced by the bacterial endotoxin lipopolysaccharide (LPS), with an efficacy comparable to that of the polyphenol compound resveratrol. Such effect was further demonstrated to occur via modulation of the NF-kappaB signaling pathway. These results indicate that bacterial metabolite, 2,3-butanediol has a negative regulatory effect on host innate immunity response, suggesting bacteria may use some metabolites for host immune evasion. PMID- 17913547 TI - In vitro cultured peripheral blood mononuclear cells from patients with chronic schistosomiasis mansoni show immunomodulation of cyclin D1,2,3 in the presence of soluble egg antigens. AB - Infection with Schistosoma mansoni induces a wide range of effects on the immune responses of the host. In the present study we investigated the influence of soluble egg antigens (SEA) on the cell cycle of peripheral blood mononuclear cells (PBMC) from infected and non-infected individuals with S. mansoni resident in an endemic area and blood donors from non-endemic area. The cell cycle, the expression of activation markers and cyclin D(+)(1,2,3) CD3(+) frequency was assessed by flow cytometry. Stimulation of PBMC from infected patients with SEA resulted in a lower frequency of CD3(+) T cells in S phase when compared with the non-infected group. In addition, infected patients presented a decrease of activation marker expression (CD69(+), HLA-DR(+) and CD28(-) on CD4(+) cells and CD25(+), HLA-DR(+) on CD8(+) cells). A reduced frequency was observed of cyclin D(1,2,3) expression in SEA-stimulated T cells from infected individuals when compared with those from the non-infected group. The decreased expression of activation markers and frequency of cyclin D(1,2,3) in T cells may result in arrest of T cells in the G(0)/G(1) phase of the cell cycle, thus explaining the down-regulation observed in chronic schistosomiasis. PMID- 17913549 TI - Cardiovascular disease in patients with spondyloarthropathies. AB - Spondyloarthropathies are associated with a greater cardiovascular risk than expected based on the cardiac lesions known to occur in these diseases. The prevalence of several conventional risk factors is high in spondyloarthropathy patients, and chronic inflammation also contributes to premature plaque formation. In addition, susceptibility genes for spondyloarthropathies may be associated with an increased risk of cardiovascular disease. Finally, several drugs used to treat spondyloarthropathies may contribute to the occurrence of cardiovascular events. A careful evaluation of the cardiovascular risk profile is a key component of the management of patients with spondyloarthropathies. PMID- 17913550 TI - Aromatase inhibitor-associated arthralgias: Pathogenesis, frequency and management. PMID- 17913551 TI - Is dynamic exercise beneficial in patients with rheumatoid arthritis? AB - INTRODUCTION: Dynamic exercise therapy as defined by the American College of Sports Medicine for healthy individuals is of unclear relevance to patients with rheumatoid arthritis (RA). No recommendations on this issue are available. Few studies have evaluated the optimal program, frequency, or target population; furthermore, there is no consensus about the best assessment tools for monitoring clinical, functional, and structural parameters during dynamic exercise therapy in patients with RA. METHODS: We conducted an extensive review of the literature published between 1964 and 2005. We identified nine randomized controlled studies that provided a high level of proof regarding the effects of dynamic exercise therapy in RA patients older than 18 years of age. RESULTS: Dynamic exercise programs improve aerobic capacity and muscle strength in patients with RA. Their effects on functional capacity are unclear, and many sources of bias influenced the study results. The clinical and laboratory safety profiles were good. The structural impact of dynamic exercise remains to be determined. PMID- 17913552 TI - Get moving! Dynamic exercise therapy for rheumatoid arthritis. PMID- 17913553 TI - Efficacy of pamidronate in erosive degenerative disk disease: A pilot study. PMID- 17913554 TI - Impact of etanercept on the costs of rheumatoid arthritis (RA): results from a French observational study. AB - INTRODUCTION: Economical impact of rheumatoid arthritis (RA) has been widely modified thanks to TNF inhibitors. Our study aims to estimate the impact etanercept prescription, in term of health resources consumption, within a regional cohort of French RA patients. METHODS: The study included 148 RA patients, with a mean follow-up duration of 343 days before and after etanercept initiation. Data were anonymously collected from ERASME database of French Health Insurance in Midi-Pyrenees region. A patient-by-patient microcosting approach was performed. RESULTS: The average annual cost per patient, attributable to RA, was 2.8 times higher after treatment by etanercept than before (15,148.57euro versus 5248.95euro). We observed a rise in pharmaceutical costs, from 11.7% of direct medical costs before to 69.7% after etanercept initiation (120.12euro versus 9995.23euro). We observed a small decrease particularly for NSAIDs (142.14euro versus 102.21euro) and physiotherapy (286.40euro versus 138.77euro). Attributable act costs and indirect costs did not differ before and after etanercept initiation. DISCUSSION: In this short-term study, initiation of etanercept in RA patients did not come along with a decrease of consumption of health resources. Long-term studies are needed to reveal a potential economical advantage as a consequence of the clinical, structural and functional efficacy of anti-TNF. PMID- 17913555 TI - Burkholderia pseudomallei - a rare cause for septic arthritis. PMID- 17913556 TI - Prolonged inflammatory syndrome revealing asymptomatic Staphylococcus cohnii infection of spinal fixation material. PMID- 17913557 TI - [Carpal dislocation combined with a lunate fracture. Report of a case ]. AB - A fracture of the semi-lunar in the frontal plan seems to be a rare and still exceptional hurt when it accompanies a retro-lunar dislocation of carp. Our case allows to see again the lesionnel mechanism of the dislocations of carp particularly in its retro-lunar variety and so to classify our case or rather to individualize it and to put it there "except series". PMID- 17913558 TI - [The return of the GAS Mask]. PMID- 17913559 TI - A treadmill control protocol combining nonlinear, equally smooth increases in speed and gradient: exercise testing for subjects with gait and exercise limitations. AB - Incremental exercise testing with a linear increase in work rate is the recommended method for clinical exercise testing. A recent protocol (A), incorporating a linear increase in speed and a nonlinear increase in gradient, has been developed which addresses some limitations of traditional testing methods. It does not account for those with an impaired gait pattern. We propose and assess a novel protocol (B) incorporating nonlinear increases in both speed and gradient. We theoretically develop a new treadmill control protocol (B), determine oxygen uptake response linearity, initial metabolic rate and cardiopulmonary response parameters (peak oxygen uptake, lactate threshold, dynamic O2 cost) and compare the outcome measures with two previously verified IET protocols (A and C (constant speed with linear increase in gradient)). Feasibility and outcomes were explored with a subject with incomplete spinal cord injury. The average initial metabolic rate (VO2) was substantially lower during protocol A (0.49 (+/-0.12) l min(-1)) and protocol B (0.52 (+/-0.05) l min(-1)) than during protocol C (1.35 (+/-0.04) l min(-1)). The average linearity of the VO2 response during protocols A and B (correlation co-efficients 0.97 (+/-0.00) and 0.95 (+/-0.02), and co-efficients of determination 0.94 (+/-0.01) and 0.91 (+/-0.02), respectively) were higher than during protocol C (correlation co efficient 0.91 (+/-0.02) and co-efficient of determination 0.84 (+/-0.02)). The average dynamic O2 cost for protocol C (6.53 (+/-0.46) ml min(-1)W(-1)) was lower than that of protocol A (10.02 (+/-1.16) ml min(-1) W(-1)) and protocol B (10.03 (+/-0.91) ml min(-1) W(-1)). No differences were found in these parameters between protocols A and B. The new protocol B performs better than protocol C and is comparable with protocol A. When testing subjects with an impaired gait pattern, it may be advantageous to use the new protocol B due to the gradual increases in both speed and gradient throughout the test. PMID- 17913560 TI - Impairment of complex upper limb motor function in de novo Parkinson's disease. AB - The aim of the present study was to evaluate complex upper limb motor function in newly diagnosed, untreated Parkinson's disease (PD) patients. Four different unimanual upper limb motor tasks were applied to 13 newly diagnosed, untreated PD patients and 13 age- and sex-matched controls. In a handwriting task, PD patients had significantly reduced sentence length and writing velocity, and decreasing letter height in the course of writing. Furthermore, PD patients performed an aiming task slower with than without target, and showed increased transposition in a pointing task. The results of this study extend previous observations of impaired complex upper limb movements to newly diagnosed, untreated PD patients. PMID- 17913561 TI - 1H-MRS experiences after bilateral DBS of the STN in Parkinson's disease. AB - The objective of this study was to evaluate the changes in the concentrations of certain brain metabolites in 13 patients with Parkinson's disease before and after bilateral subthalamic nucleus (STN DBS). The N-acetylaspartate (NAA)/choline (Chol), NAA/creatine (Cr), Chol/Cr ratios were determined by single voxel Proton magnetic resonance spectroscopy ((1)H-MRS) studies on 1.0T unit using short TE stimulated echo acquisition mode (STEAM) sequence. Spectra were obtained from the right and left globus pallidus, and left fronto-basal cortex. The patients were also assessed according to the UPDRS part III, in the "medication-on and off" conditions. CONCLUSIONS: after STN DBS cortical NAA/Cho, NAA/Cr ratios increased significantly, which were highly correlated with the significant improvements of the UPDRS scores. PMID- 17913562 TI - Association between depression and socio-economic status among community-dwelling elderly in Japan: the Aichi Gerontological Evaluation Study (AGES). AB - This study investigated the prevalence of depression by area and socio-economic status (SES). Study participants were 32,891 Japanese elderly aged 65 and over who responded to a postal survey conducted in 15 municipalities in 2003. Depression was assessed using a short version of the Geriatric Depression Scale (GDS-15). SES was measured using years of education and income adjusted for family size. After adjustment for age, illness, higher level of activities of daily living, sex, marital status, and self-rated health, depression was still significantly associated with lower SES and residential area. Further research should also examine the area effect on health. PMID- 17913563 TI - Longitudinal sliding of the median nerve in patients with non-specific arm pain. AB - In patients with non-specific arm pain (NSAP; also known as repetitive strain injury), there are clinical signs of altered median nerve sliding. It is possible that a restriction along the nerve course will lead to abnormal increases in local strain during limb movements, possibly contributing to symptoms. The present study uses ultrasound imaging to examine median nerve sliding through the proximal and distal nerve segments in 18 NSAP patients. Longitudinal nerve sliding was measured during metacarpophalangeal, wrist and elbow movements. During elbow movements, the angle of elbow extension at which the nerve begins to move was determined, since this was expected to decrease with a restriction through the shoulder. The results from this study were compared with previously reported data. Nerve movements ranged from 1.26 to 4.73 mm in patients compared with 1.43-5.57 mm in controls. There was no significant difference in nerve sliding (p>0.05) or in the angle of elbow extension at which the nerve began to move (mean=53.4 degrees in patients, 52.0 degrees in controls; p>0.05). In summary, restriction of median nerve sliding is unlikely to play a major role in NSAP. Therefore, painful responses during limb movements which tension the nerve are unlikely to result from abnormal increases in nerve strain. PMID- 17913564 TI - Shoulder kinematic features using arm elevation and rotation tests for classifying patients with frozen shoulder syndrome who respond to physical therapy. AB - Physical therapy is an intervention commonly used in the treatment of subjects with frozen shoulder symptoms, with limited proven effect. The purpose of this study was to identify the kinematic features of patients with frozen shoulder who are more likely to respond to physical therapy. Thirty-four subjects presenting frozen shoulder syndrome were studied to determine altered shoulder kinematics and functional disability. Subjects received the same standardized treatment with passive mobilization/stretching techniques, physical modalities (i.e. ultrasound, shortwave diathermy and/or electrotherapy) and active exercises twice a week for 3 months. Initially, subjects were asked to perform full active motion in 3 tests: abduction in the scapular plane, hand-to-neck and hand-to-scapula. During the test, shoulder kinematics were measured using a 3-D electromagnetic motion capturing system. In the initial and follow-up sessions, the self-reported Flexilevel Scale of Shoulder Function (FLEX-SF) was used to determine functional disability from symptoms. Improvement with treatment was determined using percent change in FLEX-SF scores over three months of treatment [(final score-initial score)/initial score x 100, >20% improvement and < = 20% nonimprovement]. Shoulder kinematics were first analysed for univariate accuracy in predicting improvement and then combined into a multivariate prediction method. A prediction method with two variables (scapular tipping >8.4 degrees during arm elevation, and external rotation >38.9 degrees during hand to neck) were identified. The presence of these two variables (positive likelihood ratio=15.71) increased the probability of improvement with treatment from 41% to 92%. It appears that shoulder kinematics may predict improvement in subjects with frozen shoulder syndrome. Prospective validation of the proposed prediction method is warranted. PMID- 17913565 TI - Response to: Brandt, C., Sole, G., Krause, M.W., Nel, M., 2007. An evidence-based review on the validity of the Kaltenborn rule as applied to the glenohumeral joint. Manual Therapy 12 (1), 3-11. PMID- 17913566 TI - Social neuroeconomics: the neural circuitry of social preferences. AB - Combining the methods of neuroscience and economics generates powerful tools for studying the brain processes behind human social interaction. We argue that hedonic interpretations of theories of social preferences provide a useful framework that generates interesting predictions and helps interpret brain activations involved in altruistic, fair and trusting behaviors. These behaviors are consistently associated with activation in reward-related brain areas, such as the striatum, and with prefrontal activity implicated in cognitive control, the processing of emotions, and integration of benefits and costs, consistent with resolution of a conflict between self-interest and other-regarding motives. PMID- 17913567 TI - Anatomy of deductive reasoning. AB - Much of cognitive research on deductive reasoning has been preoccupied with advocating for or against visuospatial (mental model theory) or linguistic/syntactic (mental logic theory) models of logical reasoning. Neuroimaging studies bear on this issue by pointing to both language-based and visuospatial systems being engaged during logical reasoning, and by raising additional issues not anticipated by these cognitive theories. Here, the literature on the neural basis of deductive reasoning from the past decade is reviewed. Although these results might seem chaotic and inconsistent, we identify several interesting patterns and articulate their implications for cognitive theories of reasoning. Cognitive neuroscience data point away from a unitary system for logical reasoning and towards a fractionated system dynamically reconfigured in response to specific task and environmental cues. PMID- 17913568 TI - Morphologic changes in cat epithelium following continuous wear of orthokeratology lenses: a pilot study. AB - PURPOSE: To investigate the sequence of morphologic changes over time in cat epithelium during continuous wear of orthokeratology lenses. METHODS: Four 2-year old female cats were used; one served as a no lens wear control and three wore custom designed Paragon CRT lenses for myopic and hyperopic correction in the right and left eyes, respectively. Lenses were worn continuously and animals were euthanased after 4h, 8h and 14 days. Corneal tissue was fixed then stained with hematoxylin and eosin for histologic evaluation and measurement of epithelial thickness. RESULTS: Average epithelial thickness of control eyes in the centre (38+/-1 microm) and mid-periphery (3.0 mm from the centre, 38+/-2 microm) of the cornea was similar. Epithelial thickness in myopic corrected eyes showed progressive thinning in the centre and progressive thickening in the mid periphery with increased lens wearing time. Hyperopic corrected eyes showed the opposite pattern of progressive epithelial thickening in the centre and thinning in the mid-periphery with lens wearing time. CONCLUSIONS: The epithelium appears to play a major role in the changes induced by orthokeratology lenses. The epithelial effects were dependent on time and lens design. Further studies are needed to determine the mechanisms responsible for these changes. PMID- 17913569 TI - Plant organelle proteomics. AB - It is important for cell biologists to know the subcellular localization of proteins to understand fully the functions of organelles and the compartmentation of plant metabolism. The accurate description of an organelle proteome requires the ability to identify genuine protein residents. Such accurate assignment is difficult in situations where a pure homogeneous preparation of the organelle cannot be achieved. Practical limitations in both organelle isolation and also analysis of low abundance proteins have resulted in limited datasets from high throughput proteomics approaches. Here, we discuss some examples of quantitative proteomic methods and their use to study plant organelle proteomes, with particular reference to methods designed to give unequivocal assignments to organelles. PMID- 17913571 TI - Study of preferential solvation of 2,6-diaminoanthraquinone in binary mixtures by absorption and fluorescence studies. AB - The role of solute-solvent and solvent-solvent interaction on the preferential solvation characteristics of 2,6-diaminoanthraquinone (DAAQ) has been analysed by monitoring the optical absorption and fluorescence emission spectra. Binary mixtures consist of dimethylformamide (DMF)-ethanol (EtOH), DMF dimelthylsulfoxide (DMSO), benzene (BZ)-DMF and acetonitrile (ACN)-DMF. The optical absorption spectra maximum and emission spectra maximum of DAAQ show the changes with varying the solvents and change in the composition in the case of binary mixtures. Non-ideal solvation characteristics are observed in all binary mixtures. It is found that at certain concentrations two mixed solvents interact to form a common structure with a nu(12) (wave number in cm(-1)) value not always intermediate (nu(1) and nu(2)) between the values of the solvents mixed. Synergistic effect is observed in the case of DMF-EtOH mixtures. The preferential solvation parameters local mole fraction X(2)(L), solvation index delta(S2), exchange constant K(12) are calculated in all binary mixtures expect in the case of DMF-BZ mixture and DMF-EtOH mixture in the ground state. We have also monitored excitation wavelength effect on the probe molecule in aprotic polar and protic polar solvents. PMID- 17913570 TI - Genotoxic effects of environmental estrogen-like compounds in CHO-K1 cells. AB - Some environmental estrogen-like compounds, such as bisphenol A (BPA), 4 nonylphenol (NP), 4-octylphenol (OP), propyl p-hydroxybenzoate (P-PHBA), and butyl p-hydroxybenzoate (B-PHBA), synthetic estrogen, diethylstilbestrol (DES), and natural estrogen, 17beta-estradiol (E2), were studied for their genotoxicity in CHO-K1 cells using sister-chromatid exchange (SCE), chromosome aberration (CA), and DNA strand break (comet) assays. Six of the chemicals, excluding E2, caused DNA migration in the comet assay and induced SCEs at one or more of the highest doses. Among the chemicals, OP produced an especially high incidence of SCEs. Structural CA was induced by five of the chemicals, excluding OP and NP, and BPA, E2, and DES also induced aneuploid cells. E2 and DES particularly increased the rate of polyploidy at high doses. The incidence of colchicine mitosis-like (c-mitotic) figures suggesting spindle disrupting effects was also detected with five of the chemicals, excluding OP and NP, and six of the chemicals, excluding E2, caused endoreduplication (ERD), a form of nuclear polyploidization induced by block of cell cycle at G2 phase, at one or more high doses. Our present results suggest that OP and NP cause repairable DNA damage, including SCEs, and do not result in CA, while the damage caused by DES, BPA, P PHBA, and B-PHBA results in the induction of CAs together with SCEs probably because of imperfect repair. We are unable to explain the observation that the DNA damage caused by E2 resulted in CA induction but not DNA migration or SCE induction, except for speculating that the DNA damage is different from that caused by DES and the estrogen-like chemicals. Our findings also suggest that E2, DES and BPA have aneuploidogenic properties, and that the former two of chemicals also are polyploidy-inducing agents. PMID- 17913572 TI - Synthesis, potentiometric and antimicrobial studies on metal complexes of isoxazole Schiff bases. AB - The metal complexes of Cu(II), Ni(II) and Co(II) with Schiff bases of 3-(2 hydroxy-3-ethoxybenzylideneamino)-5-methyl isoxazole [HEBMI] and 3-(2-hydroxy-5 nitrobenzylidene amino)-5-methyl isoxazole [HNBMI] which were obtained by the condensation of 3-amino-5-methyl isoxazole with substituted salicylaldehydes have been synthesized. Schiff bases and their complexes have been characterized on the basis of elemental analysis, magnetic moments, molar conductivity, thermal analysis and spectral (IR, UV, NMR and Mass) studies. The spectral data show that these ligands act in a monovalent bidentate fashion, co-ordinating through phenolic oxygen and azomethine nitrogen atoms. Chelates of Co(II), Ni(II) appear to be octahedral and Cu(II) appears to be distorted octahedral. To investigate the relationship between formation constants of binary complexes and antimicrobial activity, the dissociation constants of Schiff bases and stability constants of their binary metal complexes have been determined potentiometrically in aqueous solution at 30+/-1 degrees C and at 0.1 M KNO3 ionic strength and discussed. Antimicrobial activities of the Schiff bases and their complexes were screened. The structure-activity correlation in Schiff bases and their metal(II) complexes are discussed, based on the effect of their stability constants. It is observed that the activity enhances upon complexation and the order of activity is in accordance with stability order of metal ions. PMID- 17913573 TI - Fast quality control of Herba Epimedii by using Fourier transform infrared spectroscopy. AB - Herba Epimedii is a well-known traditional Chinese medicine (TCM) having the effect of nourishing the kidney and strengthening the 'Yang'. Its primary effective constituents are considered to be the 8-prenyl flavonols, which can be assorted into 4'-methoxyl-prenylflavonols (MPFs) and 4'-hydroxyl-prenylflavonols (HPFs), according to the group (methoxyl or hydroxyl) located at 4' in their structures. The Fourier transform infrared spectroscopy (FT-IR) has been widely used in the researches of TCMs. In the present study, the FT-IR was attempted to be applied in the quality control of Herba Epimedii. We compared the IR spectra of 17 pure flavonoids, of which eight were derived from Herba Epimedii, and found a characteristic absorption peak at 1259+/-1 cm(-1), corresponding to the MPFs, the major 8-prenyl flavonols in the aerial parts of the Epimedium species. This peak could also be found in the IR spectra of both the herbal samples and their 70% ethanol extracts. Moreover, the intensity of this peak was in the direct correlation with the total content of MPFs. The correlation values, representing the semblance of two spectra, of the IR spectrum of herbal sample and icariin, in the range of 1280-1200 cm(-1), had been established to be a good index for the quality control of the herbs. Accordingly, a correlation value of not less than 0.50 could be used as the essential screening criteria for the herbs. The FT-IR could be used for the fast and effective quality control of Herba Epimedii. PMID- 17913574 TI - Spectroscopic study of phase transitions in natural calcite mineral. AB - The process and the formation of new minerals upon heating the carbonate rocks containing clay minerals, together with calcite are determined with thermal analysis, X-ray diffraction, infrared and Raman spectroscopy. The calcite-calcium oxide phase transition sequence was followed up to 947 degrees C in naturally occurring limestone samples. The spectral variations of the internal modes of the carbonate trigonal (nu(1), nu(2), nu(3) and nu(4)) were used to probe the structural phase transitions. The calcium oxide phase (which on reaction with atmospheric water forms portlandite) with an onset temperature of around 950 degrees C was also characterized by the appearance of the infrared mode around 450 cm(-1). The minerals, which were formed upon heating the calcite, were calcium oxide and wollastonite. PMID- 17913575 TI - Mossbauer analysis of substituted diiron(II) tetraiminediphenolate macrocyclic complexes. AB - A new series of substituted diiron(II) complexes [Fe(2)(tidf)(L)(2)(MeOH)(2)](n+) (tidf=a two compartment tetraiminediphenolate macrocycle; n=0 or 2+; L=NCS(-), CN(-), N(3)(-), pyrazine (pz), 4-cyanopyridine (4-cnpy) and 4-mercaptopyridine (4 shpy)) and one tetranuclear complex, {[Fe(2)(tidf)(CH(3)OH)(2)](2)(mu-4 cnpy)(2)}(ClO(4))(4) were isolated and characterized by elemental analysis, conductivity measurements, Mossbauer and FTIR. PMID- 17913576 TI - The study on the effect and mechanism of the second ligands on the luminescence properties of terbium complexes. AB - The binary complex of Tb(III) with N-phenylanthranilic acid (N-HPA) was synthesized, and the ternary complexes were synthesized by introducing 1,10 phenanthroline (Phen), 2,2'-dipyridyl (Bipy), trioctylphosphine oxide (TPPO) as the second ligand, respectively. These complexes were characterized by infrared spectra, UV spectra and fluorescence spectra. The effect and mechanism of different second ligands on the fluorescent intensity of the terbium N phenylanthranilic acid complexes was discussed. It showed that all the complexes exhibited ligand-sensitized green emission. The luminescence intensity increased in the sequence of Tb(N-PA)(3)Phenmacrophages (LC50 approximately 29 micromol L(-1), 24 h; 39 micromol L(-1), 48 h)>skin cells (LC50 approximately 91 micromol L(-1), 24 h; approximately 66 micromol L(-1), 48 h). Paradoxically, a high concentration of resveratrol (50 micromol L(-1)) inhibited the proliferation of all three cell types, and a low concentration (5 micromol L( 1)) stimulated the proliferation of macrophages. The viability of macrophages was also decreased by piceatannol in a concentration-dependent manner. The stimulation of macrophages with zymosan lowered the cytotoxicity of both resveratrol and piceatannol. Scanning electron microscopy of cells treated with resveratrol revealed changes in cellular morphology that were consistent with toxicity. In macrophages and skin cells, resveratrol (50 micromol L(-1)) induced a time-dependent increase in reduced glutathione levels but did not alter the background levels of thiobarbituric acid-reactive substances. Taken together, the present data indicate that resveratrol is toxic to cultured macrophages, T cells and skin cells at concentrations>or=25 micromol L(-1), and that the cytotoxicity occurs via a mechanism that does not involve oxidative stress. Furthermore, the degree of toxicity of both resveratrol and piceatannol towards macrophages depends on the activation status of these cells, with zymosan-activated cells appearing more resistant than nonstimulated cells. PMID- 17913684 TI - Effect of aluminium on the levels of some essential elements in occupationally exposed workers. AB - The mechanism of aluminium-induced cytotoxicity has not yet been defined. This study investigated possible changes in essential elements in workers occupationally exposed to Al fumes. It included 60 exposed workers and a matching control group of 60 employees not occupationally exposed to Al. Mean serum copper, calcium, zinc and iron were significantly lower in the exposed group than in controls. In addition, mean plasma and urine levels of Al were significantly higher in the exposed employees than in the controls. A statistically significant negative correlation was found between plasma and urinary Al and the studied essential elements. These findings corroborate the hypothesis that Al exposure has an adverse effect on essential elements in humans, with subsequent impact on the cellular enzymatic and metabolic processes. PMID- 17913685 TI - Drug analysis in necrophagous flies and human tissues. AB - Necrophagous insects may provide useful information about the time, place and cause of death. In addition, they can serve as reliable alternative specimens for toxicological analysis in cases where human tissue and fluids, normally taken during autopsies, are not available, due to decomposition of the corpse. This paper reports the results of drug analysis of the larvae of two fly families, Calliphoridae and Sarcophagidae, collected from the body of a middle-aged man who had committed suicide approximately three weeks before his corpse was found. Multiple samples of decomposed human tissue, of the blowfly, and of the larval flesh were analysed using gas chromatography/mass spectrometry (GC/MS), and amphetamine was detected in all samples. While the screening results were beyond doubt, the quantitative analysis was less clear, and further research is needed in this area. PMID- 17913686 TI - Suicide by fentanyl. AB - Fentanyl is a potent, short-acting narcotic analgesic widely used as surgical anaesthetic. This article presents a case in which fentanyl was self-injected by a 41-year old nurse, an employee at the hospital emergency department, who was found dead at home. She had no known history of drug and alcohol abuse. Two syringes, one empty and one filled with a clear liquid, were found near the body, while a needle was stuck into her hand. Toxicological analysis showed fentanyl poisoning. Fentanyl overdose was declared the cause of death and the manner of death was classified as suicide. To our knowledge, death due to the intravenous injection of fentanyl has not previously been reported in Croatia. PMID- 17913687 TI - Fatal poisoning by alcohol and heroin. AB - Drug abuse with alcohol consumption have been on the rise in Split-Dalmatian County for a while now. This article reports two separate cases with three deaths due to fatal combinations of heroin and alcohol. The first case of poisoning is related to a young couple, a 30-year-old man and a 28-year-old woman, who were found dead in a car, surrounded by cans of a variety alcoholic drinks. Two needles were found beside the bodies as well. The victims were registered drug abusers who had been in withdrawal programs. The second case was a 29-year-old man who was found dead in a house. Three fresh injection marks were visible on his right arm, and two needles were near his body. He was not known as a drug addict, but he had tried to commit suicide recently. Carboxyhaemoglobin was found in blood samples of both victims from the first case. The concentration was 25% and that could contribute to their death. In both described cases blood alcohol concentration was higher then 1.60 g kg(-1). Toxicology tests were positive for heroin, meconin, acetaminophen, 6-acetylmorphine, codeine, noscapine and papaverine. Ethanol, being a respiratory depressant, combined with morphine drastically increases the risk of rapid death due to respiration failure. PMID- 17913688 TI - Mechanisms of organophosphate toxicity and detoxication with emphasis on studies in Croatia. AB - This review comprises studies on the mechanisms of toxicity and detoxication of organophosphorus (OP) compounds done in Croatia in different research areas. One area is the synthesis of antidotes against OP poisoning and their in vivo testing in experimental animals. In vitro studies included in this review focus on the mechanisms of reversible inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), protection of cholinesterases from inhibition by OPs, and reactivation of phosphylated cholinesterases. The third area comprises distribution profiles of BChE and paraoxonase (PON) phenotypes in selected population groups and the detection of OPs and metabolites in humans. Finally, methods are described for the detection of OP compounds in human blood and other media by means of cholinesterase inhibition. PMID- 17913689 TI - Application of recombinant DNA methods for production of cholinesterases as organophosphate antidotes and detectors. AB - To develop new avenues for synthesizing novel antidotes for organophosphate poisoning and for detection of the organophosphates, we have turned to recombinant DNA methods to synthesize cholinesterases with unusual properties. For antidotal therapy we describe mutations of the native mouse and human enzymes that allow for enhanced rates of oxime reactivation. Such enzymes, when localized in the circulation, would enable the circulating cholinesterase to become a catalytic rather than simply a stoichiometric scavenger. Hence, "oxime-assisted catalysis" provides a means for scavenging the organophosphates in the circulation thereby minimizing their tissue penetration and toxicity. Accordingly, the oxime antidote or prophylactic agent has a dual action within the circulation and at the tissue level. Second, through a novel chemistry, termed freeze-frame, click chemistry, we have used organophosphate conjugates of acetylcholinesterase as templates for the synthesis of novel nucleophilic reactivating agents. Finally, acetylcholinesterase can be modified through cysteine substitution mutagenesis and attachment of fluorophores at the substitution positions. When linked at certain locations in the molecule, the attached fluorophore is sensitive to organophosphate conjugation with acetylcholinesterase, and thus the very target of insecticide or nerve agent action becomes a detection molecule for organophosphate exposure. PMID- 17913690 TI - 3-D structure of serum paraoxonase 1 sheds light on its activity, stability, solubility and crystallizability. AB - Serum paraoxonases (PONs) exhibit a wide range of physiologically important hydrolytic activities, including drug metabolism and detoxification of nerve gases. PON1 and PON3 reside on high-density lipoprotein (HDL) (the "good cholesterol"), and are involved in the alleviation of atherosclerosis. Members of the PON family have been identified not only in mammals and other vertebrates, but also in invertebrates. We earlier described the first crystal structure of a PON family member, a directly-evolved variant of PON1, at 2.2 A resolution. PON1 is a 6-bladed beta-propeller with a unique active-site lid which is also involved in binding to HDL. The 3-D structure, taken together with directed evolution studies, permitted analysis of mutations which enhanced the stability, solubility and crystallizability of this PON1 variant. The structure permits a detailed description of PON1's active site and suggests possible mechanisms for its catalytic activity on certain substrates. PMID- 17913691 TI - Red blood cell acetylcholinesterase and plasma butyrylcholinesterase status: important indicators for the treatment of patients poisoned by organophosphorus compounds. AB - Inhibition of acetylcholinesterase (AChE) is regarded as the primary toxic mechanism of organophosphorus compounds (OP). Therapeutic strategies are directed to antagonise overstimulation of muscarinic receptors with atropine and to reactivate inhibited AChE with oximes. Reactivation is crucial within the neuromuscular synapse, where atropine is ineffective, since peripheral neuromuscular block eventually leads to respiratory failure. Patients with OP intoxication have to be identified as early as possible. During an international NBC-defence exercise anesthetised pigs were poisoned with sarin, followed by treatment with atropine and oxime. Blood samples were drawn and red blood cell (RBC)-AChE activity determined with a fielded test system on-site. Within a few minutes the poisoning was verified. After administration of HI-6, RBC-AChE activity increased rapidly. Blood samples were reanalysed in our laboratory in Munich. Almost identical course of the AChE activities was recorded by both systems.The more comprehensive cholinesterase status was determined in Munich. Oxime administration can be stopped when AChE is aged completely, but has to be continued as long as poison is present in the body and reactivation is possible. To aid the on-site physician in optimising diagnosis and treatment, a fielded test system should be available to allow rapid determination of the complete cholinesterase status. PMID- 17913692 TI - Contaminants in fish: risk-benefit considerations. AB - Fish provide a healthful source of dietary protein and are high in nutrients such as omega-3 fatty acids. There is evidence of beneficial effects of fish consumption in coronary heart disease, stroke, age-related macular degeneration, and growth and development. Yet, benefits may be offset by the presence of contaminants, such as methylmercury (MeHg), dioxins, polychlorinated biphenyls (PCBs) and several other halogenated persistent organic pollutants. MeHg is a known developmental neurotoxicant, as evidenced by several animal studies and episodes of human intoxication in Japan and Iraq. Fish represent the main source of exposure to MeHg for the general population, and large predatory fish (swordfish, tuna) have the highest levels of MeHg contamination. Provisional tolerable weekly intakes of 0.7 microg kg(-1) to 1.6 microg kg(-1) have been set by regulatory agencies. Concern for contamination of fish with dioxins and dioxin like PCBs stems from their reported carcinogenicity, immunotoxicity, and reproductive and developmental toxicities. Farmed and wild-caught fish appear to have similar levels of contaminants. Advisories are in place that recommend limited consumption of certain fish in children, pregnant women and women of childbearing age. Careful risk-benefit considerations should foster fish consumption while minimizing exposure to toxic contaminants. PMID- 17913693 TI - Female-biased dispersal and patrilocal kin groups in a mammal with resource defence polygyny. AB - In most mammals, dispersal rates are higher in males than in females. Using behavioural and genetic data of individually marked bats, we show that this general pattern is reversed in the greater sac-winged bat (Saccopteryx bilineata). Dispersal is significantly female biased and male philopatry in combination with rare male immigration causes a patrilineal colony structure. Female dispersal helps avoid father-daughter inbreeding, as male tenure exceeds female age at first breeding in this bat species. Furthermore, our data suggest that females may engage in extra-harem copulations to mate with genetically dissimilar males, and thus avoid their male descendants as mating partners. Acquaintance with the natal colony might facilitate territory takeover since male sac-winged bats queue for harem access. Given the virtual absence of male immigration and the possible lower reproductive success of dispersing males, we argue that enhancing the likelihood of settlement of male descendants could be adaptive despite local mate competition. We conclude that resource defence by males is important in promoting male philopatry, and argue that the potential overlap of male tenure and female first conception is the driving force for females to disperse. PMID- 17913694 TI - Non-determinable defibrillation threshold and inefficacy of implantable cardioverter/defibrillator shocks due to defective connections of the defibrillator lead terminals in the device header port. AB - Inefficacy of implantable cardioverter/defibrillator (ICD) shocks or inappropriate shocks in ICD patients may occur due to mechanical or technical failure of the device. In our unusual case of ICD defibrillation failure, faulty insertion and configuration of the defibrillation lead in the header port resulted in ineffective shock therapies and a non-determinable defibrillation threshold. In cases of successless defibrillations during or after ICD implantation, this problem should be considered. PMID- 17913695 TI - The quick-implantable-defibrillator trial. AB - AIMS: Earlier ICD therapy included an electrophysiological study (EPS), an extensive defibrillation threshold test (DFT), and a pre-discharge test. Now that ICD-therapy is widely accepted, an EPS is no longer performed in most patients, extensive DFT-tests have been reduced to a minimum of two effective shocks and discharge tests have been discarded in most centres. However, it has never been demonstrated prospectively that this simplification is safe. METHODS AND RESULTS: The Quick-Implantable-Defibrillator (Quick-ICD) Trial was a prospective multi centre trial, which randomized patients, who had survived a cardiac arrest (SCD) or an unstable ventricular tachycardia (VT), to two different clinical strategies: (a) The extensive strategy included an EPS, an extensive DFT-test, and a pre-discharge test; (b) In the simplified approach (quick strategy) the ICD was implanted without an EPS and a pre-discharge test. Two effective shocks during implantation at 21 J were sufficient. The primary endpoint of this trial was a cluster of adverse events related to the diagnostic approach and to ICD therapy. One hundred and ninety patients were included, 97 randomized to the extensive-, 93 to the quick strategy. Mean follow-up was 12 +/- 7 months. Twenty seven patients reached the endpoint in the quick group and 32 in the extensive group. During follow-up, the event-free survival was equal in the two study arms (test for equivalence, P = 0.0044). The initial hospital stay was significantly shorter in the quick population (8.4 +/- 4.7 vs. 11.2 +/- 7.4 days, P = 0.004) CONCLUSION: It is safe and cost-effective to implant an ICD without an EPS, an extensive DFT-, and a pre-discharge test in carefully selected patients after survived SCD or unstable VTs. PMID- 17913696 TI - What is the role of the pacing rate in the prevention of atrial tachyarrhythmias? PMID- 17913697 TI - Increased base rate of atrial pacing for prevention of atrial fibrillation after implantation of a dual-chamber pacemaker: insights from the Atrial Overdrive Pacing Study. AB - AIMS: Different pacing sites and various algorithms have been utilized to prevent atrial fibrillation (AF) in pacemaker recipients. However, the optimal pacing rate settings have not yet been established. In this randomized, prospective, multicentre, single-blinded, cross over study, rate-adaptive pacing at a high base rate (BR) in patients, age 60 years or above, or a history of paroxysmal AF, who underwent dual-chamber (DDD) pacemaker implantation for standard pacing indications, was evaluated for prevention of AF. METHODS AND RESULTS: In the study cohort of 145 patients implanted with DDD pacemakers with a programmable rest rate (RR) feature, the BR/RR settings were sequentially but randomly adjusted as follows: 60 bpm/Off for the baseline quarter (initial 3 months) and then to either 'A-B-C' or 'C-B-A' settings (A = 70/65 bpm, B = 70/Off, C = 80/65 bpm) for the subsequent quarters each of 3 months duration. Data on automatic mode switch episodes, device diagnostics, and a questionnaire evaluating pacemaker awareness and palpitations were collected. Ninety-nine patients, mean age 77 +/- 10 years, who completed the study protocol and followed for 12 months did not show significant differences in the number of mode switch episodes between any settings used. The percentage of atrial pacing was lower during baseline pacing compared to settings A, B, and C (P < 0.0001). Setting C produced a higher percentage of atrial pacing than A and B (P < 0.01). Although a higher percentage of atrial pacing correlated with a lower incidence of mode switch episodes, there was no statistically significant difference in the number of mode switch episodes between settings A, B, and C. There were no significant differences in the questionnaire scores relating to pacemaker awareness or palpitation. CONCLUSION: Overdrive single-site pacing in the right atrium achieved by programming analysed settings in the present study did not reduce AF as assessed by mode switch episodes. Additionally, no change in the symptoms of arrhythmia or awareness of pacing was seen. PMID- 17913698 TI - Catheter ablation of atrial fibrillation: do we know what we are doing? PMID- 17913699 TI - Extensive contribution of embryonic stem cells to the development of an evolutionarily divergent host. AB - The full potential of embryonic stem (ES) cells to generate precise cell lineages and complex tissues can be best realized when they are differentiated in vivo i.e. in developing blastocysts. Owing to various practical and ethical constraints, however, it is impossible to introduce ES cells of certain species into blastocysts of the same species. One solution is to introduce ES cells into blastocysts of a different species. However, it is not known whether ES cells can contribute extensively to chimerism when placed into blastocysts of a distantly related species. Here, we address this question using two divergent species, Apodemus sylvaticus and Mus musculus, whose genome sequence differs by approximately 18% from each other. Despite this considerable evolutionary distance, injection of Apodemus ES cells into Mus blastocysts led to viable chimeras bearing extensive Apodemus contributions to all major organs, including the germline, with Apodemus contribution reaching approximately 40% in some tissues. Immunostaining showed that Apodemus ES cells have differentiated into a wide range of cell types in the chimeras. Our results thus provide a proof of principle for the feasibility of differentiating ES cells into a wide range of cell types and perhaps even complex tissues by allowing them to develop in vivo in an evolutionarily divergent host-a strategy that may have important applications in research and therapy. Furthermore, our study demonstrates that mammalian evolution can proceed at two starkly contrasting levels: significant divergence in genome and proteome sequence, yet striking conservation in developmental programs. PMID- 17913700 TI - Protein phosphatase 1 binds to the RNA recognition motif of several splicing factors and regulates alternative pre-mRNA processing. AB - Alternative splicing emerges as one of the most important mechanisms to generate transcript diversity. It is regulated by the formation of protein complexes on pre-mRNA. We demonstrate that protein phosphatase 1 (PP1) binds to the splicing factor transformer2-beta1 (tra2-beta1) via a phylogenetically conserved RVDF sequence located on the RNA recognition motif (RRM) of tra2-beta1. PP1 binds directly to tra2-beta1 and dephosphorylates it, which regulates the interaction between tra2-beta1 and other proteins. Eight other proteins, including SF2/ASF and SRp30c, contain an evolutionary conserved PP1 docking motif in the beta-4 strand of their RRMs indicating that binding to PP1 is a new function of some RRMs. Reducing PP1 activity promotes usage of numerous alternative exons, demonstrating a role of PP1 activity in splice site selection. PP1 inhibition promotes inclusion of the survival of motoneuron 2 exon 7 in a mouse model expressing the human gene. This suggests that reducing PP1 activity could be a new therapeutic principle to treat spinal muscular atrophy and other diseases caused by missplicing events. Our data indicate that the binding of PP1 to evolutionary conserved motifs in several RRMs is the link between known signal transduction pathways regulating PP1 activity and pre-mRNA processing. PMID- 17913701 TI - A block of autophagy in lysosomal storage disorders. AB - Most lysosomal storage disorders (LSDs) are caused by deficiencies of lysosomal hydrolases. While LSDs were among the first inherited diseases for which the underlying biochemical defects were identified, the mechanisms from enzyme deficiency to cell death are poorly understood. Here we show that lysosomal storage impairs autophagic delivery of bulk cytosolic contents to lysosomes. By studying the mouse models of two LSDs associated with severe neurodegeneration, multiple sulfatase deficiency (MSD) and mucopolysaccharidosis type IIIA (MPSIIIA), we observed an accumulation of autophagosomes resulting from defective autophagosome-lysosome fusion. An impairment of the autophagic pathway was demonstrated by the inefficient degradation of exogenous aggregate-prone proteins (i.e. expanded huntingtin and mutated alpha-synuclein) in cells from LSD mice. This impairment resulted in massive accumulation of polyubiquitinated proteins and of dysfunctional mitochondria which are the putative mediators of cell death. These data identify LSDs as 'autophagy disorders' and suggest the presence of common mechanisms in the pathogenesis of these and other neurodegenerative diseases. PMID- 17913702 TI - Genetic determinants of emotionality and stress response in AcB/BcA recombinant congenic mice and in silico evidence of convergence with cardiovascular candidate genes. AB - Genomic loci bearing stress-related phenotypes were dissected in recombinant congenic strains (RCS) of mice with C57BL/6J (B6) and A/J progenitors. Adult male mice from 14 A/J and 22 B6 background lines were evaluated for emotional reactivity in open-field (OF) and elevated plus-maze tests. Core temperature was monitored by radio telemetry during immobilization and on standard as well as salt-enriched diets. In addition, urinary electrolytes were measured. Genome-wide linkage analysis of the parameters revealed over 20 significant quantitative trait loci (QTL). The highest logarithm of odds (LOD) scores were within the previously-reported OF emotionality locus on Chr 1 (LOD = 4.6), in the dopa decarboxylase region on Chr 11 for the plus-maze (LOD = 4.7), and within a novel region of calmodulin 1 on Chr 12 for Ca++ excretion after a 24-h salt load (LOD = 4.6). RCS stress QTL overlapped with several candidate loci for cardiovascular (CV) disease. In silico evidence of functional polymorphisms by comparative sequence analysis of progenitor strains assisted to ascertain this convergence. The anxious BcA70 strain showed down regulation of Atp1a2 gene expression in the heart (P < 0.001) and brain (P < 0.05) compared with its parental B6 strain, compatible with the enhanced emotionality described in knock out animals for this gene, also involved in the salt-sensitive component of hypertension. Functional polymorphisms in regulatory elements of candidate genes of the CV/inflammatory/immune systems support the hypothesis of genetically-altered environmental susceptibility in CV disease development. PMID- 17913703 TI - Mitochondrial thymidine kinase and the enzymatic network regulating thymidine triphosphate pools in cultured human cells. AB - In non-proliferating cells mitochondrial (mt) thymidine kinase (TK2) salvages thymidine derived from the extracellular milieu for the synthesis of mt dTTP. TK2 is a synthetic enzyme in a network of cytosolic and mt proteins with either synthetic or catabolic functions regulating the dTTP pool. In proliferating cultured cells the canonical cytosolic ribonucleotide reductase (R1-R2) is the prominent synthetic enzyme that by de novo synthesis provides most of dTTP for mt DNA replication. In non-proliferating cells p53R2 substitutes for R2. Catabolic enzymes safeguard the size of the dTTP pool: thymidine phosphorylase by degradation of thymidine and deoxyribonucleotidases by degradation of dTMP. Genetic deficiencies in three of the participants in the network, TK2, p53R2, or thymidine phosphorylase, result in severe mt DNA pathologies. Here we demonstrate the interdependence of the different enzymes of the network. We quantify changes in the size and turnover of the dTTP pool after inhibition of TK2 by RNA interference, of p53R2 with hydroxyurea, and of thymidine phosphorylase with 5 bromouracil. In proliferating cells the de novo pathway dominates, supporting large cytosolic and mt dTTP pools, whereas TK2 is dispensable, even in cells lacking the cytosolic thymidine kinase. In non-proliferating cells the small dTTP pools depend on the activities of both R1-p53R2 and TK2. The activity of TK2 is curbed by thymidine phosphorylase, which degrades thymidine in the cytoplasm, thus limiting the availability of thymidine for phosphorylation by TK2 in mitochondria. The dTTP pool shows an exquisite sensitivity to variations of thymidine concentrations at the nanomolar level. PMID- 17913704 TI - TNFR1 and TNFR2 signaling interplay in cardiac myocytes. AB - Tumor necrosis factor alpha (TNFalpha) plays a major role in chronic heart failure, signaling through two different receptor subtypes, TNFR1 and TNFR2. Our aim was to further delineate the functional role and signaling pathways related to TNFR1 and TNFR2 in cardiac myocytes. In cardiac myocytes isolated from control rats, TNFalpha induced ROS production, exerted a dual positive and negative action on [Ca(2+)] transient and cell fractional shortening, and altered cell survival. Neutralizing anti-TNFR2 antibodies exacerbated TNFalpha responses on ROS production and cell death, arguing for a major protective role of the TNFR2 pathway. Treatment with either neutralizing anti-TNFR1 antibodies or the glutathione precursor, N-acetylcysteine (NAC), favored the emergence of TNFR2 signaling that mediated a positive effect of TNFalpha on [Ca(2+)] transient and cell fractional shortening. The positive effect of TNFalpha relied on TNFR2 dependent activation of the cPLA(2) activity, independently of serine 505 phosphorylation of the enzyme. Together with cPLA(2) redistribution and AA release, TNFalpha induced a time-dependent phosphorylation of ERK, MSK1, PKCzeta, CaMKII, and phospholamban on the threonine 17 residue. Taken together, our results characterized a TNFR2-dependent signaling and illustrated the close interplay between TNFR1 and TNFR2 pathways in cardiac myocytes. Although apparently predominant, TNFR1-dependent responses were under the yoke of TNFR2, acting as a critical limiting factor. In vivo NAC treatment proved to be a unique tool to selectively neutralize TNFR1-mediated effects of TNFalpha while releasing TNFR2 pathways. PMID- 17913705 TI - Molecular proximity of cystic fibrosis transmembrane conductance regulator and epithelial sodium channel assessed by fluorescence resonance energy transfer. AB - We present the evidence for a direct physical association of cystic fibrosis transmembrane conductance regulator (CFTR) and epithelial sodium channel (ENaC), two major ion channels implicated in the pathophysiology of cystic fibrosis, a devastating inherited disease. We employed fluorescence resonance energy transfer, a distance-dependent imaging technique with capability to detect molecular complexes with near angstrom resolution, to estimate the proximity of CFTR and ENaC, an essential variable for possible physical interaction to occur. Fluorescence resonance energy transfer studies were complemented with a classic biochemical approach: coimmunoprecipitation. Our results place CFTR and ENaC within reach of each other, suggestive of a direct interaction between these two proteins. PMID- 17913706 TI - Myc down-regulation as a mechanism to activate the Rb pathway in STAT5A-induced senescence. AB - Senescence is a general antiproliferative program that avoids the expansion of cells bearing oncogenic mutations. We found that constitutively active STAT5A (ca STAT5A) can induce a p53- and Rb-dependent cellular senescence response. However, ca-STAT5A did not induce p21 and p16(INK4a), which are responsible for inhibiting cyclin-dependent protein kinases and engaging the Rb pathway during the senescence response to oncogenic ras. Intriguingly, ca-STAT5A led to a down regulation of Myc and Myc targets, including CDK4, a negative regulator of Rb. The down-regulation of Myc was in part proteasome-dependent and correlated with its localization to promyelocytic leukemia bodies, which were found to be highly abundant during STAT5-induced senescence. Introduction of CDK4 or Myc bypassed STAT5A-induced senescence in cells in which p53 was also inactivated. These results uncover a novel mechanism to engage the Rb pathway in oncogene-induced senescence and indicate the existence of oncogene-specific pathways that regulate senescence. PMID- 17913707 TI - Identification of in vivo phosphorylation sites and their functional significance in the sodium iodide symporter. AB - The Na+/I- symporter (NIS)-mediated iodide uptake activity is the basis for targeted radioiodide ablation of thyroid cancers. Although it has been shown that NIS protein is phosphorylated, neither the in vivo phosphorylation sites nor their functional significance has been reported. In this study, Ser-43, Thr-49, Ser-227, Thr-577, and Ser-581 were identified as in vivo NIS phosphorylation sites by mass spectrometry. Kinetic analysis of NIS mutants of the corresponding phosphorylated amino acid residue indicated that the velocity of iodide transport of NIS is modulated by the phosphorylation status of Ser-43 and Ser-581. We also found that the phosphorylation status of Thr-577 may be important for NIS protein stability and that the phosphorylation status of Ser-227 is functionally silent. Thr-49 appears to be critical for proper local structure/conformation of NIS because mutation of Thr-49 to alanine, aspartic acid, or serine results in reduced NIS activity without alterations in total or cell surface NIS protein levels. Taken together, we showed that NIS protein levels and functional activity could be modulated by phosphorylation through distinct mechanisms. PMID- 17913708 TI - Generation of an analog-sensitive Syk tyrosine kinase for the study of signaling dynamics from the B cell antigen receptor. AB - The Syk protein-tyrosine kinase is an essential component of the signaling machinery that couples the B cell receptor for antigen to multiple downstream signal transduction pathways. Syk is phosphorylated and activated rapidly and transiently following receptor engagement, but many signaling events, such as the activation of transcription factors occur over the course of several minutes or hours. To investigate a role for the continued activation of Syk in these processes, we generated an analog-sensitive mutant with an engineered ATP-binding pocket to render the kinase uniquely sensitive to an orthogonal inhibitor. Mutation of the gatekeeper residue in Syk yielded an enzyme with very low activity. Second-site mutations, selected based on structural comparisons between Syk and Src, were introduced that restored catalytic activity to the mutant Syk. Syk-deficient DT40 B cells were prepared expressing the analog-sensitive Syk (Syk AQL). Inhibition of the activity of Syk prior to, concomitant with or shortly following receptor engagement led to the rapid inhibition of receptor-mediated tyrosine phosphorylation and blocked the activation of extracellular signal regulated kinase, NF-kappaB, and NFAT. The receptor-mediated activation of NF kappaB required active Syk for a relatively short period of time, whereas the activation of NFAT required active kinase for a prolonged (>1 h) period. Receptor cross-linking led to the recruitment of Syk to the clustered receptor. Retention of these receptor-kinase complexes on the cell surface was dependent on the continued activity of Syk. Thus, despite the apparent transient nature of the activation of Syk, the catalytic activity of the Syk was required for sustained signaling from ligated receptors. PMID- 17913709 TI - Regulation of Nox1 activity via protein kinase A-mediated phosphorylation of NoxA1 and 14-3-3 binding. AB - Nox activator 1 (NoxA1) is a homologue of p67(phox) that acts in conjunction with Nox organizer 1 (NoxO1) to regulate reactive oxygen species (ROS) production by the NADPH oxidase Nox1. The phosphorylation of cytosolic regulatory components by multiple kinases plays important roles in assembly and activity of the phagocyte NADPH oxidase (Nox2) system, but little is known about regulation by phosphorylation in the Nox1 system. Here we identify Ser(172) and Ser(461) of NoxA1 as phosphorylation sites for protein kinase A (PKA). A consequence of this phosphorylation was the enhancement of NoxA1 complex formation with 14-3-3 proteins. Using both a transfected human embryonic kidney 293 cell Nox1 model system and endogenous Nox1 in colon cell lines, we showed that the elevation of cAMP inhibits, whereas the inhibition of PKA enhances, Nox1-dependent ROS production through effects on NoxA1. Inhibition of Nox1 activity was intensified by the availability of 14-3-3zeta protein, and this regulatory interaction was dependent on PKA-phosphorylatable sites at Ser(172) and Ser(461) in NoxA1. We showed that phosphorylation and 14-3-3 binding induce the dissociation of NoxA1 from the Nox1 complex at the plasma membrane, suggesting a mechanism for the inhibitory effect on Nox1 activity. Our data establish that PKA-phosphorylated NoxA1 is a new binding partner of 14-3-3 protein(s) and that this forms the basis of a novel mechanism regulating the formation of ROS by Nox1 and, potentially, other NoxA1-regulated Nox family members. PMID- 17913710 TI - Oxidative modification to cysteine sulfonic acid of Cys111 in human copper-zinc superoxide dismutase. AB - Copper-zinc superoxide dismutase (SOD1) plays a protective role against oxidative stress. On the other hand, recent studies suggest that SOD1 itself is a major target of oxidative damage and has its own pathogenicity in various neurodegenerative diseases, including familial amyotrophic lateral sclerosis. Only human and great ape SOD1s among mammals have the highly reactive free cysteine residue, Cys(111), at the surface of the SOD1 molecule. The purpose of this study was to investigate the role of Cys(111) in the oxidative damage of the SOD1 protein, by comparing the oxidative susceptibility of recombinant human SOD1 modified with 2-mercaptoethanol at Cys(111) (2-ME-SOD1) to wild-type SOD1. Wild type SOD1 was more sensitive to oxidation by hydrogen peroxide-generating fragments, oligomers, and charge isomers compared with 2-ME-SOD1. Moreover, wild type SOD1, but not 2-ME-SOD1, generated an upper shifted band in reducing SDS PAGE even by air oxidation. Using mass spectrometry and limited proteolysis, this upper band was identified as an oxidized subunit of SOD1; the sulfhydryl group (Cys-SH) of Cys(111) was selectively oxidized to cysteine sulfinic acid (Cys SO(2)H) and to cysteine sulfonic acid (Cys-SO(3)H). The antibody raised against a synthesized peptide containing Cys(111)-SO(3)H reacted with only the Cys(111) peroxidized SOD1 by Western blot analysis and labeled Lewy body-like hyaline inclusions and vacuole rims in the spinal cord of human SOD1-mutated amyotrophic lateral sclerosis mice by immunohistochemical analysis. These results suggest that Cys(111) is a primary target for oxidative modification and plays an important role in oxidative damage to human SOD1, including familial amyotrophic lateral sclerosis mutants. PMID- 17913711 TI - Structural basis for the regulation of N-acetylglutamate kinase by PII in Arabidopsis thaliana. AB - PII is a highly conserved regulatory protein found in organisms across the three domains of life. In cyanobacteria and plants, PII relieves the feedback inhibition of the rate-limiting step in arginine biosynthesis catalyzed by N acetylglutamate kinase (NAGK). To understand the molecular structural basis of enzyme regulation by PII, we have determined a 2.5-A resolution crystal structure of a complex formed between two homotrimers of PII and a single hexamer of NAGK from Arabidopsis thaliana bound to the metabolites N-acetylglutamate, ADP, ATP, and arginine. In PII, the T-loop and Trp(22) at the start of the alpha1-helix, which are both adjacent to the ATP-binding site of PII, contact two beta-strands as well as the ends of two central helices (alphaE and alphaG) in NAGK, the opposing ends of which form major portions of the ATP and N-acetylglutamate substrate-binding sites. The binding of Mg(2+).ATP to PII stabilizes a conformation of the T-loop that favors interactions with both open and closed conformations of NAGK. Interactions between PII and NAGK appear to limit the degree of opening and closing of the active-site cleft in opposition to a domain separating inhibitory effect exerted by arginine, thus explaining the stimulatory effect of PII on the kinetics of arginine-inhibited NAGK. PMID- 17913712 TI - Crystal structure of an unusual thioredoxin protein with a zinc finger domain. AB - Many Gram-negative bacteria have two cytoplasmic thioredoxins, thioredoxin-1 and 2, encoded by the trxA and trxC genes, respectively. Both thioredoxins have the highly conserved WCGPC motif and function as disulfide-bond reductases. However, thioredoxin-2 has unique features: it has an N-terminal motif that binds a zinc ion, and its transcription is under the control of OxyR, which allows it to be up regulated under oxidative stress. Here, we report the crystal structure of thioredoxin-2 from Rhodobacter capsulatus. The C-terminal region of thioredoxin-2 forms a canonical thioredoxin fold with a central beta-sheet consisting of five strands and four flanking alpha-helices on either side. The N-terminal zinc finger is composed of four short beta-strands (S1-S4) connected by three short loops (L1-L3). The four cysteines are at loops L1 and L3 and form a tetragonal binding site for a zinc ion. The zinc finger is close to the first beta-strand and first alpha-helix of the thioredoxin fold. Nevertheless, the zinc finger may not directly affect the oxidoreductase activity of thioredoxin-2 because the zinc finger is not near the active site of a protomer and because thioredoxin-2 is a monomer in solution. On the basis of structural similarity to the zinc fingers in Npl4 and Vps36, we propose that the N-terminal zinc finger of thioredoxin-2 mediates protein-protein interactions, possibly with its substrates or chaperones. PMID- 17913713 TI - Expression of bisecting type and Lewisx/Lewisy terminated N-glycans on human sperm. AB - Human sperm lack major histocompatibility class I molecules, making them susceptible to lysis by natural killer (NK) cells. Major histocompatibility class I negative tumor cells block NK cell lysis by expressing sufficient amounts of bisecting type N-glycans on their surfaces. Therefore, sperm could employ the same strategy to evade NK cell lysis. The total N-glycans derived from sperm were sequenced using ultrasensitive mass spectrometric and conventional approaches. Three major classes of N-glycans were detected, (i) high mannose, (ii) biantennary bisecting type, and (iii) biantennary, triantennary, and tetraantennary oligosaccharides terminated with Lewisx and Lewisy sequences. Immunostaining of normal sperm showed that glycoproteins bearing Lewisy sequences are localized to the acrosome and not the plasma membrane. In contrast, defective sperm showed distinct surface labeling with anti-Lewisy antibody. The substantial expression of high mannose and complex type N-glycans terminated with Lewisx and Lewisy sequences suggests that sperm glycoproteins are highly decorated with ligands for DC-SIGN. Based on previous studies, the addition of such carbohydrate signals should inhibit antigen-specific responses directed against sperm glycoproteins in both the male and female reproductive systems. Thus, the major N glycans of human sperm are associated with the inhibition of both innate and adaptive immune responses. These results provide more support for the eutherian fetoembryonic defense system hypothesis that links the expression of carbohydrate functional groups to the protection of gametes and the developing human in utero. This study also highlights the usefulness of glycomic profiling for revealing potential physiological functions of glycans expressed in specific cell types. PMID- 17913714 TI - Different roles of alpha- and beta-branch xanthophylls in photosystem assembly and photoprotection. AB - Xanthophylls (oxygenated carotenoids) are essential components of the plant photosynthetic apparatus, where they act in photosystem assembly, light harvesting, and photoprotection. Nevertheless, the specific function of individual xanthophyll species awaits complete elucidation. In this work, we analyze the photosynthetic phenotypes of two newly isolated Arabidopsis mutants in carotenoid biosynthesis containing exclusively alpha-branch (chy1chy2lut5) or beta-branch (chy1chy2lut2) xanthophylls. Both mutants show complete lack of qE, the rapidly reversible component of nonphotochemical quenching, and high levels of photoinhibition and lipid peroxidation under photooxidative stress. Both mutants are much more photosensitive than npq1lut2, which contains high levels of viola- and neoxanthin and a higher stoichiometry of light-harvesting proteins with respect to photosystem II core complexes, suggesting that the content in light-harvesting complexes plays an important role in photoprotection. In addition, chy1chy2lut5, which has lutein as the only xanthophyll, shows unprecedented photosensitivity even in low light conditions, reduced electron transport rate, enhanced photobleaching of isolated LHCII complexes, and a selective loss of CP26 with respect to chy1chy2lut2, highlighting a specific role of beta-branch xanthophylls in photoprotection and in qE mechanism. The stronger photosystem II photoinhibition of both mutants correlates with the higher rate of singlet oxygen production from thylakoids and isolated light-harvesting complexes, whereas carotenoid composition of photosystem II core complex was not influential. In depth analysis of the mutant phenotypes suggests that alpha branch (lutein) and beta-branch (zeaxanthin, violaxanthin, and neoxanthin) xanthophylls have distinct and complementary roles in antenna protein assembly and in the mechanisms of photoprotection. PMID- 17913715 TI - Prevalence and characterization of integrons from bacteria isolated from a slaughterhouse wastewater treatment plant. AB - OBJECTIVES: To investigate the presence and distribution of integron-carrying bacteria from a slaughterhouse wastewater treatment plant (WWTP). METHODS: Enterobacteriaceae and aeromonads were isolated at different stages of the wastewater treatment process and screened for the presence of integrase genes by dot-blot hybridization. Integrase-positive strains were characterized in terms of phylogenetic affiliation, genetic content of integrons and antimicrobial resistance profiles. Plasmid location of some integrons was established by Southern-blot hybridization. Strains containing integron-carrying plasmids were selected for mating experiments. RESULTS: Integrase genes were present in all samples, including the final effluent. The global prevalence was determined to be 35%, higher than in other aquatic environments. Forty-two integrase-positive isolates were further characterized. Nine distinct cassette arrays were found, containing genes encoding resistance to beta-lactams (bla(OXA-30)), aminoglycosides (aadA1, aadA2, aadA13, aadB), streptothricin (sat1, sat2), trimethoprim (dfrA1, dfrA12), a putative esterase (estX) and a protein with unknown function (orfF). Gene cassette arrays aadA1, dfrAI-aadA1 and estX-sat2 aadA1 were common to aeromonads and Enterobacteriaceae. The class 2 integron containing an estX-sat2-aadA1 cassette array was detected for the first time in Aeromonas sp. Nearly 12% (5 out of 43) of intI genes were located in plasmids. intI genes from isolates MM.1.3 and MM.1.5 were successfully conjugated into Escherichia coli at frequencies of 3.79 x 10(-5) and 5.46 x 10(-5) per recipient cell, respectively. CONCLUSIONS: Our data support the hypothesis that WWTPs constitute a potential hot spot for horizontal gene transfer and for selection of antimicrobial resistance genes among aquatic bacteria. Moreover, water discharges represent a possible risk for dissemination of undesirable genetic traits. PMID- 17913716 TI - The Candida glabrata putative sterol transporter gene CgAUS1 protects cells against azoles in the presence of serum. AB - OBJECTIVES: The uptake of endogenous sterol from serum may allow Candida glabrata to survive azole treatment. This study aims to determine the contribution of a sterol transporter that alters fluconazole sensitivity in the presence of serum. METHODS: Bioinformatic analysis predicted CgAUS1 as the C. glabrata orthologue of the Saccharomyces cerevisiae transporters AUS1 and PDR11. To investigate whether the CgAUS1 gene has sterol transporter activity, we investigated the effects of an AUS1 deletion on the growth of a tetracycline-regulatable ERG9 strain (tet ERG9aus1), wherein ERG9 expression is turned off giving rise to a sterol requirement. Tetracycline-dependent repression of CgAUS1 in the tet-AUS1 strain was used to determine the fluconazole susceptibility of CgAUS1 in the presence and absence of serum. RESULTS: The tetracycline-treated tet-ERG9aus1 strain failed to grow in the presence of serum, whereas the parental tet-ERG9AUS1 strain grew by incorporating sterol from exogenously supplied serum. Serum cholesterol protected cells against the antifungal effects of fluconazole and this protection was lost by repressing CgAUS1 gene expression. Furthermore, such protection was also observed during itraconazole treatment, but not observed in cells treated with non-azole antifungals. CONCLUSIONS: CgAUS1 appears to function as a sterol transporter that may contribute to lower azole susceptibility in the presence of serum and to protect C. glabrata against azole toxicity in vivo. PMID- 17913717 TI - High diversity of extended-spectrum beta-lactamases among clinical isolates of Enterobacteriaceae from Portugal. AB - OBJECTIVES: To investigate the occurrence and the diversity of Ambler class A ESBLs among Enterobacteriaceae from different Portuguese clinical settings over a 2 year period (2002-04). METHODS: One hundred and nine extended-spectrum beta lactamase (ESBL)-producing Enterobacteriaceae isolates from five geographically distant health institutions in Portugal were studied. ESBLs were characterized by isoelectric focussing, PCR and further sequencing. Antibiotic susceptibility testing, transfer of resistance genes and clonal diversity were determined by standard procedures. Plasmid relatedness was established by comparison of random amplified polymorphic DNA (RAPD) patterns. RESULTS: ESBLs were identified as TEM (46%), SHV (30%), CTX-M (22%) and GES (2%) types; TEM-24, TEM-52, SHV-12 and CTX M-15 enzymes being the most frequently found. Inter-hospital dissemination of epidemic strains harbouring the most prevalent ESBLs was detected, including the TEM-24-producing Enterobacter aerogenes European epidemic clone. Conjugative transfer of ESBLs was achieved for 67% of isolates and epidemic plasmids containing specific bla genes were detected (bla(CTX-M-15) and bla(TEM-24)). We describe two new ESBLs, SHV-90 (A187T, G238S and E240K) and SHV-91 (P20S and E240K), and a new TEM-type enzyme conferring a phenotype resembling that of a complex mutant TEM beta-lactamase, designated as TEM-154 (M69L and R164S). The broad-spectrum beta-lactamases SHV-26, SHV-36 and TEM-110 were first observed in our country. CONCLUSIONS: We describe a complex ESBL epidemiology in Portugal, including widespread dissemination of known strains and plasmids coding for TEM 24 and CTX-M-15 enzymes as observed in other European countries. PMID- 17913718 TI - Comment on: Impact of antibiotic resistance and of adequate empirical antibiotic treatment in the prognosis of patients with Escherichia coli bacteraemia. PMID- 17913719 TI - Evaluation of the chromogenic Cica-beta-Test for detecting extended-spectrum, AmpC and metallo-beta-lactamases. AB - BACKGROUND: Extended-spectrum, metallo- and AmpC beta-lactamases usually are sought subsequently to susceptibility testing, meaning that producers are not identified until 72 h after a clinical specimen is taken. Chromogenic tests might usefully shorten this delay, and we investigated the Cica-beta-Test for this purpose. METHODS: Reference and clinical strains with known beta-lactamases, or controls, were grown with a cefpodoxime disc to promote conservation of resistance. The cultures were then tested with nitrocefin and with the Cica-beta Test, which examines for hydrolysis of the chromogenic oxyimino-cephalosporin HMRZ-86 with and without specific inhibitors of extended-spectrum, metallo- and AmpC beta-lactamases. RESULTS: were scored, as colour changes from yellow to red, with the tester blinded to the strain identity and the mechanism(s) present. Results Proportions of extended-spectrum, metallo- and AmpC beta-lactamase producers correctly identified by the Cica-beta-Test were 85%, 77% and 72%, respectively. Such performance should be achievable if testing colonies from a primary culture plate, 24 h after a specimen was taken. Greater precision, albeit at more delay, would be achieved if results were read in conjunction with antibiogram data available 48 h after the specimen was taken. Limitations were frequent confusion of Klebsiella oxytoca hyperproducing K1 enzyme with AmpC hyperproducers, and that isolates with NMC-A or KPC carbapenemases were wrongly inferred to have AmpC enzymes. CONCLUSIONS: The Cica-beta-Test has the potential to provide useful therapeutic guidance, identifying isolates with potent beta lactamases and informing early therapy; it will also help to monitor beta lactamase epidemiology among multiresistant strains. PMID- 17913721 TI - Treatment of children persistently infected with hepatitis B virus: seroconversion or suppression. AB - We have reviewed the current strategies regarding the treatment of persistent hepatitis B virus (HBV) in children and compared these with adult strategies. The options for achieving suppression of viral DNA replication versus hepatitis B e antigen to antibody seroconversion have been evaluated. The results of studies in different geographical locations have been confounded by HBV genotypes, as it is now clear that some genotypes respond better to treatment than others. Consideration needs to be given as to whether optimal treatment strategies developed for adults are directly applicable to children. In children, early seroconversion to allow improved long-term outcomes should be considered rather than embarking on the long-term complexities of managing patients on a combination of antiviral drugs to achieve viral suppression. PMID- 17913720 TI - Linezolid versus vancomycin for the treatment of infections caused by methicillin resistant Staphylococcus aureus in Japan. AB - OBJECTIVES: To compare the efficacy and safety of linezolid and vancomycin for the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections in Japan. METHODS: Patients with nosocomial pneumonia, complicated skin and soft tissue infections or sepsis caused by MRSA were randomized to receive linezolid (600 mg every 12 h) or vancomycin (1 g every 12 h). RESULTS: One hundred patients received linezolid and 51 received vancomycin with outcomes evaluated at the end of therapy (EOT) and at the follow-up (FU), 7-14 days later. At EOT, clinical success rates in the MRSA microbiologically evaluable population were 62.9% and 50.0% for the linezolid and vancomycin groups, respectively; and microbiological eradication rates were 79.0% and 30.0% in the two groups, respectively (P < 0.0001). At FU, the clinical success rates were 36.7% for both groups and the microbiological eradication rates were 46.8% and 36.7%, respectively. Reversible anaemia (13%) and thrombocytopenia (19%) were reported more frequently in linezolid patients; laboratory analysis showed mild decrease in platelet counts with full recovery by FU. The mean platelet count in linezolid patients with thrombocytopenia was 101,000/mm(3). Significantly low platelet counts (<50,000/mm(3)) were observed more frequently in patients receiving vancomycin than in linezolid patients (6% versus 3%). Mean changes in haemoglobin levels between the two groups were not different. CONCLUSIONS: Linezolid is as effective as vancomycin for the treatment of MRSA infections and may be more effective than vancomycin in achieving microbiological eradication. Haematological adverse events were reported more frequently in linezolid-treated patients; analysis of laboratory data showed a mild reversible trend towards lower platelet counts. PMID- 17913722 TI - Mathematical modelling response of Pseudomonas aeruginosa to meropenem. AB - OBJECTIVES: Widespread emergence of resistance to antimicrobial agents is a serious problem. The rate at which new agents are made available clinically is unlikely to keep up with these resistant pathogens, and there is an urgent need to accelerate antimicrobial agent development. We explored the use of mathematical modelling to guide selection of dosing regimens. METHODS: Using time kill studies data of Pseudomonas aeruginosa over 24 h, we developed a mathematical model to capture the dynamic relationship between a heterogeneous microbial population and meropenem concentrations. The microbial behaviour in response to meropenem over 5 days was predicted via computer simulation and subsequently validated using an in vitro hollow fibre infection model. Three parallel differential equations were used, each characterizing the rate of change of drug concentration, microbial susceptibility and microbial burden of the surviving population over time, respectively. Several model structures were explored; they differed in the adaptation of the microbial population over time. Various fluctuating concentration-time profiles of meropenem were experimentally examined, mimicking human elimination and repeated dosing. RESULTS: Using limited experimental data as inputs, the mathematical model was reasonable in qualitatively predicting microbial response (sustained suppression or regrowth due to resistance emergence) to various pharmacokinetic profiles of meropenem. CONCLUSIONS: Our results suggest that mathematical modelling may be used to predict microbial response to a large number of antimicrobial agent dosing regimens efficiently, and have the potential to be used to guide highly targeted investigation of dosing regimens in pre-clinical studies and clinical trials. The in vivo relevance of the modelling approach warrants further investigations. PMID- 17913723 TI - Prevalence of etravirine (TMC-125) resistance mutations in HIV-infected patients with prior experience of non-nucleoside reverse transcriptase inhibitors. PMID- 17913724 TI - Prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in invasive isolates from southern and eastern Mediterranean countries. AB - OBJECTIVES: Efforts aimed at curtailing the ever increasing spread of methicillin resistant Staphylococcus aureus (MRSA) require effective information of its epidemiology. However, knowledge about the situation in southern and eastern countries of the Mediterranean is incomplete since reports have been sporadic and difficult to compare. METHODS: Over a 36 month period from 2003 to 2005, the ARMed project collected more than 5000 susceptibility test results of invasive isolates of S. aureus from blood cultures routinely processed within participating laboratories servicing 62 hospitals situated in Algeria, Cyprus, Egypt, Jordan, Lebanon, Malta, Morocco, Tunisia and Turkey. RESULTS: Overall, the median MRSA proportion was 39% (interquartile range: 27.1% to 51.1%). The highest proportions of MRSA were reported by Jordan, Egypt and Cyprus, where more than 50% of the invasive isolates were methicillin-resistant. Considerable variation was identified in the proportion of MRSA in hospitals within the same country. CONCLUSIONS: It appears that most of the countries in the Mediterranean region are experiencing a surge in MRSA infections. This requires a greater focus to identify relevant drivers of resistance and implement effective practices in order to address them, especially improved infection control and antibiotic consumption practices. PMID- 17913725 TI - Interactive effects of nutrient and mechanical stresses on plant morphology. AB - BACKGROUND AND AIMS: Plant species frequently encounter multiple stresses under natural conditions, and the way they cope with these stresses is a major determinant of their ecological breadth. The way mechanical (e.g. wind, current) and resource stresses act simultaneously on plant morphological traits has been poorly addressed, even if both stresses often interact. This paper aims to assess whether hydraulic stress affects plant morphology in the same way at different nutrient levels. METHODS: An examination was made of morphological variations of an aquatic plant species growing under four hydraulic stress (flow velocity) gradients located in four habitats distributed along a nutrient gradient. Morphological traits covering plant size, dry mass allocation, organ water content and foliage architecture were measured. KEY RESULTS: Significant interactive effects of flow velocity and nutrient level were observed for all morphological traits. In particular, increased flow velocity resulted in size reductions under low nutrient conditions, suggesting an adaptive response to flow stress (escape strategy). On the other hand, moderate increases in flow velocity resulted in increased size under high nutrient conditions, possibly related to an inevitable growth response to a higher nutrient supply induced by water renewal at the plant surface. For some traits (e.g. dry mass allocation), a consistent sense of variation as a result of increasing flow velocity was observed, but the amount of variation was either reduced or amplified under nutrient-rich compared with nutrient-poor conditions, depending on the traits considered. CONCLUSIONS: These results suggest that, for a given species, a stress factor may result, in contrasting patterns and hence strategies, depending on a second stress factor. Such results emphasize the relevance of studies on plant responses to multiple stresses for understanding the actual ecological breadth of species. PMID- 17913726 TI - Vegetative storage protein in Litchi chinensis, a subtropical evergreen fruit tree, possesses trypsin inhibitor activity. AB - BACKGROUND AND AIMS: Vegetative storage proteins (VSPs) are commonly bioactive in herbaceous plants but few VSPs with bioactivity have been identified in trees. In addition, information on the characterization of VSPs in evergreen trees is limited. The objective of this study was to characterize the VSPs with bioactivity in evergreen trees. Methods The VSP in lychee (Litchi chinensis), an evergreen fruit tree, was characterized by a combination of cytological, biochemical and molecular biological techniques. KEY RESULTS: The VSP in lychee was a 22-kDa protein. It accumulated in the large central vacuoles of protein storing cells (PSCs) in two distinguishable forms, granular and floccular. The PSCs were of a novel type. The 22-kDa protein is distributed in mature leaves, bark tissues of branches, trunk and large roots, paralleling the distribution of PSCs. Its homologues were present in mature seed. During young shoot development and fruiting, the 22-kDa protein decreased apparently, suggesting a nitrogen storage function. The 22-kDa protein had several isoforms encoded by a small multigene family. One gene member, LcVSP1, was cloned. The LcVSP1 had no intron and contained a 675 bp open reading frame encoding a putative protein of 225 amino acids. LcVSP1 was homologous to Kunitz trypsin inhibitors. The 22-kDa protein inhibited trypsin and chymotrypsin, but had no inhibitory effect on subtilisin. CONCLUSIONS: Lychee is rich in a 22-kDa VSP with trypsin inhibitor activity. The VSP plays an important role in nitrogen storage while its possible defensive function remains to be elucidated. PMID- 17913727 TI - Cryptic speciation in the Caesalpinia hintonii complex (Leguminosae: Caesalpinioideae) in a seasonally dry Mexican forest. AB - BACKGROUND AND AIMS: [corrected] The Caesalpinia hintonii group comprises six species of endemic shrubs or trees, C. epifanioi, C. hintonii, C. laxa, C. macvaughii, C. melanadenia and C. oyamae, found in scattered patches of seasonally dry forest in the Rio Balsas depression and the neighbouring Tehuacan Cuicatlan valley, which are part of the Mexican morphotectonic province of Sierra Madre del Sur. An evaluation is made of phylogeographic patterns and genetic diversity with a phylogenetic analysis of the C. hintonii complex in order to study the dynamics of speciation in this endemic group of legumes. METHODS: A phylogeographic study based on four highly variable non-coding plastid regions (trnL intron, trnL-F intergenic spacer, trnH-psbA intergenic spacer, and accD psaI intergenic spacer) was carried out for the Caesalpinia hintonii complex. Five of the six taxa of the C. hintonii complex were included. KEY RESULTS AND CONCLUSIONS: The plastid analyses involving multiple accessions of each taxon from throughout their ranges resolved C. epifanioi and C. hintonii as well supported clusters, but C. oyamae has two unexpectedly divergent lineages. Two well-supported geographic clades: eastern (C. epifanioi, C. melanadenia and C. oyamae) and western (C. hintonii and C. macvaughii) were established. The analyses performed provide evidence of recent morphostatic radiation in C. oyamae resulting from isolation and local adaptation. This pattern of genetic differentiation without morphological divergence may be a model that fits many groups of tropical woody taxa inhabiting similarly dry forests subjected to shifting selection. PMID- 17913728 TI - Unexpected basis for impaired Glc3Man9GlcNAc2-P-P-dolichol biosynthesis by elevated expression of GlcNAc-1-P transferase. AB - GlcNAc-1-P transferase (GPT) transfers GlcNAc-1-P from UDP-GlcNAc to dolichol-P (Dol-P), forming GlcNAc-P-PDol to initiate synthesis of the lipid-linked oligosaccharide Glc3Man9GlcNAc2-P-P-dolichol (G3M9Gn2-P-P-Dol). Elevated expression of GPT in CHO-K1 cells is known to cause accumulation of the intermediate M5Gn2-P-P-Dol, presumably by excessively consuming Dol-P and thereby hindering Dol-P-dependent synthesis of Man-P-Dol (MPD) and Glc-P-Dol (GPD), which provide the residues for extending M5Gn2-P-P-Dol to G3M9Gn2-P-P-Dol. If so, elevated GPT expression should increase oligosaccharide-P-P-Dol quantities and reduce monosaccharide-P-Dol quantities, while requiring GPT enzymatic activity. Here we report that elevated GPT expression failed to appreciably alter the quantities of the two classes of dolichol-linked saccharide, and that neither a GPT inhibitor nor introduction of an inactivating mutation into GPT prevented M5Gn2-P-P-Dol accumulation,arguing against excessive Dol-P consumption. Unexpectedly,we noticed similarities between the phenotypes of GPT overexpressers and of CHO-K1 cells lacking Lec35p (encoded by MPDU1, the congenital disorder of glycosylation(CDG)-If locus), which is required for utilization of MPD and GPD. By compensatory overexpression of Lec35p, G3M9Gn2-P-P-Dol synthesis in GPT overexpressers could be restored. However, GPT overexpression did not affect the levels of Lec35 mRNA or protein. These results suggest that GPT may impair Lec35p function, and imply that upper as well as lower limits on GPT expression exist in normal cells. Since the mammalian GPT gene can undergo spontaneous amplification, the data also indicate a potential basis for forms of pseudo-CDG-If. PMID- 17913729 TI - Reduced alpha4beta1 integrin/VCAM-1 interactions lead to impaired pre-B cell repopulation in alpha 1,6-fucosyltransferase deficient mice. AB - Mice with a targeted gene disruption of Fut8 (Fut8(-/-)) showed an abnormality in the transition from pro-B cell to pre-B cell, reduced peripheral B cells, and a decreased immunoglobulin production. Alpha 1,6-fucosyltransferase (FUT8) is responsible for the alpha 1,6 core fucosylation of N-glycans, which could modify the functions of glycoproteins. The loss of a core fucose in both very late antigen 4 (VLA-4, alpha4beta1 integrin) and vascular cell adhesion molecule 1 (VCAM-1) led to a decreased binding between pre-B cells and stromal cells, which impaired pre-B cells generation in Fut8(-/-) mice. Moreover, the B lineage genes, such as CD79a, CD79b, Ebf1, and Tcfe2a, were downregulated in Fut8(-/-) pre-B cells. Indeed, the frequency of preBCR(+)CD79b(low) cells in bone marrow pre-B cells in Fut8(-/-) was much lower than that in Fut8(+/+) cells. These results reveal a new role of core fucosylated N-glycans in mediating early B cell development and functions. PMID- 17913731 TI - Aristolochic acid: the common culprit of Chinese herbs nephropathy and Balkan endemic nephropathy. PMID- 17913730 TI - Alpha-interferon therapy for chronic hepatitis C may induce acute allograft rejection in kidney transplant patients with failed allografts. AB - BACKGROUND: In hepatitis C virus (HCV) positive kidney transplant (KT) patients, the use of alpha-interferon (alphaIFN) is contraindicated due to the risk of acute rejection (AR). Conversely, if these HCV(+) KT patients lose their allograft, re-transplantation might be contemplated provided alphaIFN therapy has been attempted. METHODS: Between 01/01/1989 and 31/12/1994, 261 kidney transplantations were performed; of these 174 were HCV(-) (group I) and 87 were HCV(+) (group II). RESULTS: At last follow-up (2006), in group I, the number of patients with a functioning graft, the number of patients who died with a functioning graft, and the number of patients who lost their graft before or after month (M) 12 were 92 (52.8%), 14 (8%), 20 (11.5%) and 48 (27.7%), respectively. In group II, the corresponding figures were 22 (25.3%; P < 0.0001), 8 (9.1%; ns), 9 (10.3%; ns) and 48 (55.3%; P < 0.0001). In group I, 19 of 48 (39.5%) patients with failed allografts after M12 underwent transplantectomy (TX) compared to 14 of 48 (29%; ns) in group II. In group II, 11 of 48 (23%) patients were offered alphaIFN therapy after their allograft failed: of these, four (36.3%) developed AR during alphaIFN therapy leading to TX. Histology, in addition to chronic allograft lesions, showed acute cellular and vascular lesions. In patients who were not offered alphaIFN therapy, TX was performed less frequently, i.e. in only six cases (16.2%). CONCLUSIONS: We conclude that even alphaIFN-treated KT patients with a failed allograft can experience acute allograft rejection that requires transplantectomy during therapy. PMID- 17913732 TI - Sodium citrate anticoagulation during sustained low efficiency dialysis (SLED) in patients with acute renal failure and severely impaired liver function. PMID- 17913733 TI - Hyponatremia associated with sodium valproate in a 22-year-old male. PMID- 17913734 TI - Circulating endothelial cells in renal disease: markers and mediators of vascular damage. PMID- 17913735 TI - Renal cyst infection in autosomal dominant polycystic kidney disease. PMID- 17913736 TI - Fluoridation of drinking water and chronic kidney disease: absence of evidence is not evidence of absence. PMID- 17913737 TI - Glomerular filtration rate--what is the rationale and justification of normalizing GFR for body surface area? PMID- 17913738 TI - Influenza vaccination does not result in an increase in relapses in patients with ANCA-associated vasculitis. AB - BACKGROUND: Vaccination against influenza has been suggested to induce relapses of ANCA-associated vasculitis but evidence is lacking. In this study, we assessed whether vaccination against influenza increases the occurrence of relapses in patients with ANCA-associated vasculitis. METHODS: Two hundred and thirty consecutive patients with ANCA-associated vasculitis from our out-patient clinics of a tertiary referral center, with at least 1 year of follow-up, were included. Retrospectively, the relapse rate per 100 patients at risk in patients who had been vaccinated against influenza within the preceding year and in patients who not had been vaccinated within that time period were calculated. RESULTS: The relapse rate per 100 patients at risk was lower in patients who had been vaccinated against influenza (3.4) than in patients who had not been vaccinated (6.3), when analyzed for the entire year and for every quarter of the year. Also, the disease-free survival per separate year according to the vaccination status was lower in all 5 years in patients who had been vaccinated, being statistically significant in 2 years. CONCLUSION: Vaccination against influenza does not increase the relapse rate in patients with ANCA-associated vasculitis. PMID- 17913739 TI - NORINE: a database of nonribosomal peptides. AB - Norine is the first database entirely dedicated to nonribosomal peptides (NRPs). In bacteria and fungi, in addition to the traditional ribosomal proteic biosynthesis, an alternative ribosome-independent pathway called NRP synthesis allows peptide production. It is performed by huge protein complexes called nonribosomal peptide synthetases (NRPSs). The molecules synthesized by NRPS contain a high proportion of nonproteogenic amino acids. The primary structure of these peptides is not always linear but often more complex and may contain cycles and branchings. In recent years, NRPs attracted a lot of attention because of their biological activities and pharmacological properties (antibiotic, immunosuppressor, antitumor, etc.). However, few computational resources and tools dedicated to those peptides have been available so far. Norine is focused on NRPs and contains more than 700 entries. The database is freely accessible at http://bioinfo.lifl.fr/norine/. It provides a complete computational tool for systematic study of NRPs in numerous species, and as such, should permit to obtain a better knowledge of these metabolic products and underlying biological mechanisms, and ultimately to contribute to the redesigning of natural products in order to obtain new bioactive compounds for drug discovery. PMID- 17913740 TI - LATERAL ORGAN BOUNDARIES defines a new family of DNA-binding transcription factors and can interact with specific bHLH proteins. AB - Conserved in a variety of evolutionarily divergent plant species, LOB DOMAIN (LBD) genes define a large, plant-specific family of largely unknown function. LBD genes have been implicated in a variety of developmental processes in plants, although to date, relatively few members have been assigned functions. LBD proteins have previously been predicted to be transcription factors, however supporting evidence has only been circumstantial. To address the biochemical function of LBD proteins, we identified a 6-bp consensus motif recognized by a wide cross-section of LBD proteins, and showed that LATERAL ORGAN BOUNDARIES (LOB), the founding member of the family, is a transcriptional activator in yeast. Thus, the LBD genes encode a novel class of DNA-binding transcription factors. Post-translational regulation of transcription factors is often crucial for control of gene expression. In our study, we demonstrate that members of the basic helix-loop-helix (bHLH) family of transcription factors are capable of interacting with LOB. The expression patterns of bHLH048 and LOB overlap at lateral organ boundaries. Interestingly, the interaction of bHLH048 with LOB results in reduced affinity of LOB for the consensus DNA motif. Thus, our studies suggest that bHLH048 post-translationally regulates the function of LOB at lateral organ boundaries. PMID- 17913741 TI - DOMINE: a database of protein domain interactions. AB - DOMINE is a database of known and predicted protein domain interactions compiled from a variety of sources. The database contains domain-domain interactions observed in PDB entries, and those that were predicted by eight different computational approaches. DOMINE contains a total of 20 513 unique domain-domain interactions among 4036 Pfam domains, out of which 4349 are inferred from PDB entries and 17 781 were predicted by at least one computational approach. This database will serve as a valuable resource to those working in the field of protein and domain interactions. DOMINE may not only serve as a reference to experimentalists who test for new protein and domain interactions, but also offers a consolidated dataset for analysis by bioinformaticians who seek to test ideas regarding the underlying factors that control the topological structure of interaction networks. DOMINE is freely available at http://domine.utdallas.edu. PMID- 17913742 TI - Changes in DNA repair during aging. AB - DNA is a precious molecule. It encodes vital information about cellular content and function. There are only two copies of each chromosome in the cell, and once the sequence is lost no replacement is possible. The irreplaceable nature of the DNA sets it apart from other cellular molecules, and makes it a critical target for age-related deterioration. To prevent DNA damage cells have evolved elaborate DNA repair machinery. Paradoxically, DNA repair can itself be subject to age related changes and deterioration. In this review we will discuss the changes in efficiency of mismatch repair (MMR), base excision repair (BER), nucleotide excision repair (NER) and double-strand break (DSB) repair systems during aging, and potential changes in DSB repair pathway usage that occur with age. Mutations in DNA repair genes and premature aging phenotypes they cause have been reviewed extensively elsewhere, therefore the focus of this review is on the comparison of DNA repair mechanisms in young versus old. PMID- 17913743 TI - CoVDB: a comprehensive database for comparative analysis of coronavirus genes and genomes. AB - The recent SARS epidemic has boosted interest in the discovery of novel human and animal coronaviruses. By July 2007, more than 3000 coronavirus sequence records, including 264 complete genomes, are available in GenBank. The number of coronavirus species with complete genomes available has increased from 9 in 2003 to 25 in 2007, of which six, including coronavirus HKU1, bat SARS coronavirus, group 1 bat coronavirus HKU2, groups 2c and 2d coronaviruses, were sequenced by our laboratory. To overcome the problems we encountered in the existing databases during comparative sequence analysis, we built a comprehensive database, CoVDB (http://covdb.microbiology.hku.hk), of annotated coronavirus genes and genomes. CoVDB provides a convenient platform for rapid and accurate batch sequence retrieval, the cornerstone and bottleneck for comparative gene or genome analysis. Sequences can be directly downloaded from the website in FASTA format. CoVDB also provides detailed annotation of all coronavirus sequences using a standardized nomenclature system, and overcomes the problems of duplicated and identical sequences in other databases. For complete genomes, a single representative sequence for each species is available for comparative analysis such as phylogenetic studies. With the annotated sequences in CoVDB, more specific blast search results can be generated for efficient downstream analysis. PMID- 17913744 TI - Involvement of phage phi29 DNA polymerase and terminal protein subdomains in conferring specificity during initiation of protein-primed DNA replication. AB - To initiate phi29 DNA replication, the DNA polymerase has to form a complex with the homologous primer terminal protein (TP) that further recognizes the replication origins of the homologous TP-DNA placed at both ends of the linear genome. By means of chimerical proteins, constructed by swapping the priming domain of the related phi29 and GA-1 TPs, we show that DNA polymerase can form catalytically active heterodimers exclusively with that chimerical TP containing the N-terminal part of the homologous TP, suggesting that the interaction between the polymerase TPR-1 subdomain and the TP N-terminal part is the one mainly responsible for the specificity between both proteins. We also show that the TP N terminal part assists the proper binding of the priming domain at the polymerase active site. Additionally, a chimerical 29 DNA polymerase containing the GA-1 TPR 1 subdomain could use GA-1 TP, but only in the presence of phi29 TP-DNA as template, indicating that parental TP recognition is mainly accomplished by the DNA polymerase. The sequential events occurring during initiation of bacteriophage protein-primed DNA replication are proposed. PMID- 17913745 TI - Escherichia coli RNA polymerase-associated SWI/SNF protein RapA: evidence for RNA directed binding and remodeling activity. AB - Helicase-like SWI/SNF proteins are present in organisms belonging to distant kingdoms from bacteria to humans, indicating that they perform a very basic and ubiquitous form of nucleic acid management; current studies associate the activity of SWI/SNF proteins with remodeling of DNA and DNA-protein complexes. The bacterial SWI/SNF homolog RapA-an integral part of the Escherichia coli RNA polymerase complex-has been implicated in remodeling post-termination DNA-RNA polymerase-RNA ternary complexes (PTC), however its explicit nucleic acid substrates and mechanism remain elusive. Our work presents evidence indicating that RNA is a key substrate of RapA. Specifically, the formation of stable RapA RNA intermediates in transcription and other, independent lines of evidence presented herein indicate that RapA binds and remodels RNA during transcription. Our results are consistent with RapA promoting RNA release from DNA-RNA polymerase-RNA ternary complexes; this process may be accompanied by the destabilization of non-canonical DNA-RNA complexes (putative DNA-RNA triplexes). Taken together, our data indicate a novel RNA remodeling activity for RapA, a representative of the SWI/SNF protein superfamily. PMID- 17913746 TI - The ePHD protein SPBP interacts with TopBP1 and together they co-operate to stimulate Ets1-mediated transcription. AB - SPBP (Stromelysin-1 PDGF responsive element binding protein) is a ubiquitously expressed 220 kDa nuclear protein shown to enhance or repress the transcriptional activity of various transcription factors. A yeast two-hybrid screen, with the extended plant homeodomain (ePHD) of SPBP as bait, identified TopBP1 (topoisomerase II beta-binding protein 1) as a candidate interaction partner of SPBP. TopBP1 has eight BRCA1 carboxy-terminal (BRCT) domains and is involved in DNA replication, DNA damage responses and in the regulation of gene expression. The interaction between SPBP and TopBP1 was confirmed in vitro and in vivo, and was found to be mediated by the ePHD domain of SPBP and the BRCT6 domain of TopBP1. Both SPBP and TopBP1 enhanced the transcriptional activity of Ets1 on the c-myc P1P2- and matrix metalloproteinase-3 (MMP3) promoters. Together they displayed a more than additive effect. Both proteins were associated with these promoters. The involvement of TopBP1 was dependent on the serine 1159 phosphorylation site, known to be important for transcriptional activation. Depletion of endogenous SPBP by siRNA treatment reduced MMP3 secretion by 50% in phorbol ester-stimulated human fibroblasts. Taken together, our results show that TopBP1 and SPBP interact physically and functionally to co-operate as co activators of Ets1. PMID- 17913747 TI - Phosphorylated serine 28 of histone H3 is associated with destabilized nucleosomes in transcribed chromatin. AB - Histone modifications and variants have key roles in the activation and silencing of genes. Phosphorylation of histone H3 at serine 10 and serine 28 is involved in transcriptional activation of genes responding to stress or mitogen-stimulated signaling pathways. The distribution of H3-modified isoforms in G0 phase chicken erythrocyte chromatin was investigated. H3 phosphorylated at serine 28 was found highly enriched in the active/competent gene fractions, as was H3 di- and trimethylated at lysine 4. The H3 variant H3.3 in this chromatin fraction was preferentially phosphorylated at serine 28. Conversely, H3 phosphorylated at serine 10 was present in all chromatin fractions, while H3 dimethylated at lysine 9 was associated with the chromatin-containing repressed genes. H3 phosphorylated at serine 28 was located at the promoter region of the transcriptionally active, but not competent, histone H5 and beta-globin genes. We provide evidence that H3.3 phosphorylated at serine 28 was present in labile nucleosomes. We propose that destabilized nucleosomes containing H3.3 phosphorylated at serine 28 aid in the dynamic disassembly-assembly of nucleosomes in active promoters. PMID- 17913749 TI - The Ser176 of T4 endonuclease IV is crucial for the restricted and polarized dC specific cleavage of single-stranded DNA implicated in restriction of dC containing DNA in host Escherichia coli. AB - Endonuclease (Endo) IV encoded by denB of bacteriophage T4 is an enzyme that cleaves single-stranded (ss) DNA in a dC-specific manner. Also the growth of dC substituted T4 phage and host Escherichia coli cells is inhibited by denB expression presumably because of the inhibitory effect on replication of dC containing DNA. Recently, we have demonstrated that an efficient cleavage by Endo IV occurs exclusively at the 5'-proximal dC (dC1) within a hexameric or an extended sequence consisting of dC residues at the 5'-proximal and the 3' proximal positions (dCs tract), in which a third dC residue within the tract affects the polarized cleavage and cleavage rate. Here we isolate and characterize two denB mutants, denB(W88R) and denB(S176N). Both mutant alleles have lost the detrimental effect on the host cell. Endo IV(W88R) shows no enzymatic activity (<0.4% of that of wild-type Endo IV). On the other hand, Endo IV(S176N) retains cleavage activity (17.5% of that of wild-type Endo IV), but has lost the polarized and restricted cleavage of a dCs tract, indicating that the Ser176 residue of Endo IV is implicated in the polarized cleavage of a dCs tract which brings about a detrimental effect on the replication of dC-containing DNA. PMID- 17913748 TI - Poly(ADP-ribosyl)ation in mammalian ageing. AB - Poly(ADP-ribose) polymerases (PARPs) catalyze the post-translational modification of proteins with poly(ADP-ribose). Two PARP isoforms, PARP-1 and PARP-2, display catalytic activity by contact with DNA-strand breaks and are involved in DNA base excision repair and other repair pathways. A body of correlative data suggests a link between DNA damage-induced poly(ADP-ribosyl)ation and mammalian longevity. Recent research on PARPs and poly(ADP-ribose) yielded several candidate mechanisms through which poly(ADP-ribosyl)ation might act as a factor that limits the rate of ageing. PMID- 17913750 TI - Human telomere, oncogenic promoter and 5'-UTR G-quadruplexes: diverse higher order DNA and RNA targets for cancer therapeutics. AB - Guanine-rich DNA sequences can form G-quadruplexes stabilized by stacked G-G-G-G tetrads in monovalent cation-containing solution. The length and number of individual G-tracts and the length and sequence context of linker residues define the diverse topologies adopted by G-quadruplexes. The review highlights recent solution NMR-based G-quadruplex structures formed by the four-repeat human telomere in K(+) solution and the guanine-rich strands of c-myc, c-kit and variant bcl-2 oncogenic promoters, as well as a bimolecular G-quadruplex that targets HIV-1 integrase. Such structure determinations have helped to identify unanticipated scaffolds such as interlocked G-quadruplexes, as well as novel topologies represented by double-chain-reversal and V-shaped loops, triads, mixed tetrads, adenine-mediated pentads and hexads and snap-back G-tetrad alignments. The review also highlights the recent identification of guanine-rich sequences positioned adjacent to translation start sites in 5'-untranslated regions (5' UTRs) of RNA oncogenic sequences. The activity of the enzyme telomerase, which maintains telomere length, can be negatively regulated through G-quadruplex formation at telomeric ends. The review evaluates progress related to ongoing efforts to identify small molecule drugs that bind and stabilize distinct G quadruplex scaffolds associated with telomeric and oncogenic sequences, and outlines progress towards identifying recognition principles based on several X ray-based structures of ligand-G-quadruplex complexes. PMID- 17913752 TI - Human diseases of telomerase dysfunction: insights into tissue aging. AB - There are at least three human diseases that are associated with germ-line mutations of the genes encoding the two essential components of telomerase, TERT and TERC. Heterozygous mutations of these genes have been described for patients with dyskeratosis congenita, bone marrow failure and idiopathic pulmonary fibrosis. In this review, we will detail the clinical similarities and difference of these diseases and review the molecular phenotypes observed. The spectrum of mutations in TERT and TERC varies for these diseases and may in part explain the clinical differences observed. Environmental insults and genetic modifiers that accelerate telomere shortening and increase cell turnover may exaggerate the effects of telomerase haploinsufficiency, contributing to the variability of age of onset as well as tissue-specific organ pathology. A central still unanswered question is whether telomerase dysfunction and short telomeres are a much more prominent factor than previously suspected in other adult-onset, age-related diseases. Understanding the biological effects of these mutations may ultimately lead to novel treatments for these patients. PMID- 17913751 TI - DNA damage, cellular senescence and organismal ageing: causal or correlative? AB - Cellular senescence has long been used as a cellular model for understanding mechanisms underlying the ageing process. Compelling evidence obtained in recent years demonstrate that DNA damage is a common mediator for both replicative senescence, which is triggered by telomere shortening, and premature cellular senescence induced by various stressors such as oncogenic stress and oxidative stress. Extensive observations suggest that DNA damage accumulates with age and that this may be due to an increase in production of reactive oxygen species (ROS) and a decline in DNA repair capacity with age. Mutation or disrupted expression of genes that increase DNA damage often result in premature ageing. In contrast, interventions that enhance resistance to oxidative stress and attenuate DNA damage contribute towards longevity. This evidence suggests that genomic instability plays a causative role in the ageing process. However, conflicting findings exist which indicate that ROS production and oxidative damage levels of macromolecules including DNA do not always correlate with lifespan in model animals. Here we review the recent advances in addressing the role of DNA damage in cellular senescence and organismal ageing. PMID- 17913753 TI - The ageing mitochondrial genome. AB - The population of elderly individuals has increased significantly over the past century and is predicted to rise even more rapidly in the future. Ageing is a major risk factor for many diseases such as neurodegenerative disease, diabetes and cancer. This highlights the importance of understanding the mechanisms involved in the ageing process. One plausible mechanism for ageing is accumulation of mutations in the mitochondrial genome. In this review, we discuss some of the most convincing data surrounding age-related mtDNA mutations and the evidence that these mutations contribute to the ageing process. PMID- 17913754 TI - Sevoflurane preconditioning at 1 MAC only provides limited protection in patients undergoing coronary artery bypass surgery: a randomized bi-centre trial. AB - BACKGROUND: Volatile agents can mimic ischaemic preconditioning leading to a decrease in myocardial infarct size. The present study investigated if a 15 min sevoflurane administration before cardiopulmonary bypass (CPB) has a cardioprotective effect in patients undergoing coronary surgery. METHODS: Seventy two patients were randomized in two centres. The intervention group (S) received 1 MAC sevoflurane administrated via the ventilator for 15 min followed by a 15 min washout before CPB, the control group did not. The primary outcome was the postoperative troponin Ic peak. A biopsy of the atrium was taken during canulation for enzyme dosages. Results are expressed as mean (SD). RESULTS: Neither troponin Ic nor tissular enzyme measurement exhibited any difference between the groups: peak of troponin Ic was 4.4 (5.6) in S group vs 5.2 (6.6) ng ml(-1) in control group (ns). Intratissular ecto-5'-nucleotidase activity was 7.1 (4.3) vs 8.5 (11.9), protein kinase C activity was 27.1 (15.7) vs 29.2 (28.7), tyrosine kinase activity was 101 (54.1) vs 98.5 (63.3), and P38 MAPKinase activity was 131.1 (76.1) vs 127.1 (86.8) nmol mg protein(-1) min(-1) in S group and control group, respectively (ns). However there were fewer patients with low postoperative cardiac index in S group (11% in S vs 35% in control group, P < 0.05) when considering the per protocol population. In S group, 25% of patients required an inotropic support during the postoperative period, vs 36% of patients in control group (ns). CONCLUSIONS: This study did not show a significant preconditioning signal after 15 min of sevoflurane administration. The 15 min duration might be too short or the concentration of sevoflurane too low to induce cardioprotection detected by troponin I levels. PMID- 17913755 TI - Why involve older people in research? PMID- 17913756 TI - HDL-cholesterol and physical performance: results from the ageing and longevity study in the sirente geographic area (ilSIRENTE Study). AB - BACKGROUND: High-density lipoprotein (HDL) cholesterol has been hypothesised to be a reliable marker of frailty and poor prognosis among the oldest elderly. We evaluate the relationship of HDL-cholesterol with measures of physical performance, muscle strength, and functional status in older persons aged 80years or older. METHODS: Data are from baseline evaluation of the ageing and longevity study in the Sirente geographic area (ilSIRENTE study) (n = 364). Physical performance was assessed using the physical performance battery score [short physical performance battery (SPPB)], which is based on three-timed tests: 4-m walking-speed, balance, and chair-stand tests. Muscle strength was measured by hand-grip strength. Analyses of covariance were performed to evaluate the relationship of different HDL-cholesterol levels with physical function. RESULTS: In the unadjusted analyses, physical function (as measured by the 4-m walking speed, theSPPB score, the basic and instrumental activities of daily living scales scores), but not hand-grip strength, improved significantly as HDL cholesterol tertiles increased. After adjustment for potential confounders, which included age, gender, living alone, alcohol abuse, physical activity, congestive heart failure, diabetes, cerebrovascular diseases, osteoarthritis, albumin, urea, C-reactive protein and LDL cholesterol, the association of HDL-cholesterol tertiles with the 4-m walking-speed and the SPPB score was still consistent. CONCLUSION: The present study suggests that among very old subjects living in the community the higher levels of HDL-cholesterol are associated with better functional performance. PMID- 17913758 TI - Is the degree of cognitive impairment in patients with Alzheimer's disease related to their capacity to appoint an enduring power of attorney? AB - BACKGROUND: Clinicians are often asked to retrospectively assess a patient's capacity to create an Enduring Power of Attorney (EPA). This study will investigate if capacity to create an EPA is significantly related to the degree of cognitive impairment in Alzheimer patients, and whether Mini Mental State Examination score (MMSE) is a good predictor of a patient's capacity. It also considers if socio-demographic factors are related to a patient's capacity to create an EPA. METHODS: Participants with a DSM-IV diagnosis of Alzheimer's disease were recruited from the Old Age Psychiatric service at the Queen Elizabeth Psychiatric Hospital, Birmingham, UK. A cognitive assessment of each patient was performed using the MMSE, followed by two independent assessments of their capacity to create an EPA made using a semi-structured interview. RESULTS: There was a significant association between level of cognitive impairment and capacity to create an EPA: chi(2) = 35.15 (P<0.0001). MMSE score significantly predicted capacity status (OR=1.6, 95% CI 0.863-0.979). Optimal sensitivity (86.6%, CI 67.4-95.5%) and specificity (82.2% CI 67.4-91.5%) were obtained using a cutoff MMSE score of 18. Positive predictive value (PPV): 75.8% (95% CI 57 88%), negative predictive value (PNV): 90.2% (CI 76-97%). No socio-demographic factors were significantly associated with capacity to create an EPA. CONCLUSIONS: The MMSE could be used as a screening tool to help inform a clinical capacity assessment in patients with Alzheimer's disease. It is important that patients always undergo individual clinical assessments where possible, but in situations where direct assessment is not possible MMSE score could be used to aid retrospective assessments of capacity to create an EPA. PMID- 17913757 TI - Age-related increases in DNA repair and antioxidant protection: a comparison of the Boyd Orr Cohort of elderly subjects with a younger population sample. AB - BACKGROUND: One commonly held theory of ageing is that it is caused by oxidative damage to critical molecules in the body, including proteins, lipids and nucleic acids. Accumulation of oxidative DNA damage with age will occur if there is an increase in reactive oxygen species in the body, or a decline in antioxidant defences, or a reduced efficiency of DNA repair. SUBJECTS AND METHODS: Using the comet assay, we have measured DNA breaks and oxidised purines in lymphocytes from subjects of different age groups: 20-35 (n = 40), 63-70 (n = 35), and 75-82 (n = 22). We also measured the resistance of lymphocyte DNA to H(2)O(2)-induced oxidative damage, and the repair activity of cell-free lymphocyte extracts on a substrate containing 8-oxoguanine. RESULTS: We found an increase in oxidative base damage in old age, but this apparently does not result from deterioration of either antioxidant defence or DNA repair. In fact, both of these tend to increase with age. There were few age-related differences in plasma levels of dietary antioxidants: tocopherols and retinol were higher in the older subjects, while lycopene was highest in the youngest age group. CONCLUSIONS: It is possible, that in old age, antioxidant defences and DNA repair are induced, in response to a higher level of oxidative damage, as mitochondria become more leaky and release more reactive oxygen. It is equally possible that older people, as survivors, had relatively high levels of antioxidant defences and DNA repair earlier in their lives, compared with those who did not survive to such an age. PMID- 17913759 TI - Prevalence and symptomatology of depression in older people living in institutions in England and Wales. AB - BACKGROUND: Epidemiological studies have shown that depression is common in institutional settings. However, the symptomatology of depression in this group has not been compared to those living in the community. AIMS: To estimate the prevalence of depression and depressive symptomatology in participants living in institutions and compare these to people living in other settings. METHOD: The Medical Research Council Cognitive Function and Ageing Study (MRC CFAS) is a population-based cohort comprising 13,004 individuals aged 65 and above, from five sites across England and Wales. Following screening, a stratified random sub sample of 2,640 participants received the Geriatric Mental State (GMS) examination of whom 340 resided in institutions. Diagnoses of depression were made using the Automated Geriatric Examination for Computer-assisted Taxonomy system (AGECAT). RESULTS: The prevalence of depression in those living in institutions was 27.1% (95% CI 17.8-36.3) compared to 9.3% (95% CI 7.8-10.9) in those living at home. Symptoms relating to depressed mood, severity of illness (e.g. wishing to be dead, future looking bleak) and some non-specific symptoms were more common in those living in residential homes. Depression was significantly associated with younger age (P = 0.002) and high functional disability (P = 0.009) in those living in institutions. CONCLUSIONS: Consistent with previous estimates, depression was highly prevalent in institutions, particularly in younger individuals with severe functional impairment. Those in institutions report considerably more symptoms of depression. Finding interventions which address these symptoms might improve quality of life for people in institutions, irrespective of formal diagnoses. PMID- 17913760 TI - Crohn's disease initially diagnosed after age 60 years. PMID- 17913761 TI - A cautionary tale: delayed onset rhabdomyolysis due to erythromycin/simvastatin interaction. PMID- 17913762 TI - Disequilibrium due to a vitamin B6 megadose supplement. PMID- 17913763 TI - Standards in dementia care. PMID- 17913765 TI - Percutaneous cardioplegia delivery using the miniport in minimally invasive mitral valve surgery. AB - Minimally invasive cardiac surgery involves limited exposure of cardiac structures. Extracorporeal circulation is usually conducted by peripheral cannulation. Cross-clamp can be achieved by remote ways of either balloon endoclamp or transthoracic clamp. Effective delivery of cardioplegic solution is somewhat more difficult than those above mentioned tasks. In order to prevent additional expenses, we sought to deliver cardioplegic solution in a simple, reproducible, and cost-effective way. The miniport is used for this application. The procedures are reported in detail. PMID- 17913766 TI - Spinal cord malperfusion caused by using the segmental clamp technique during descending aortic repair for chronic type B aortic dissection. AB - Several effective strategies for spinal cord protection have been advocated in descending and thoracoabdominal aortic repairs. The segmental clamp technique has been known as a useful adjunct to shorten the duration of spinal cord ischemia. However, we experienced two cases of spinal cord malperfusion during segmental aortic clamping in descending aortic repair for chronic type B aortic dissection. In these patients, the intercostal arteries including the Adamkiewicz artery had originated from the false lumen. In one patient, spinal cord ischemia was initially detected as decreased motor-evoked potentials. Transesophageal echocardiography simultaneously revealed blood flow congestion in the false lumen during segmental aortic clamping and spinal cord ischemia had developed due to malperfusion of the intercostal arteries branching from the false lumen. Segmental clamping in patients with aortic dissection may not always be useful for shortening the duration of spinal cord ischemia. Transesophageal echocardiography as well as motor-evoked potentials is a useful modality for obtaining the details of intraoperative blood flow in dissecting lumens and malperfusion of the intercostal arteries related to spinal cord injury. PMID- 17913767 TI - A balanced PGR5 level is required for chloroplast development and optimum operation of cyclic electron transport around photosystem I. AB - PSI cyclic electron transport contributes markedly to photosynthesis and photoprotection in flowering plants. Although the thylakoid protein PGR5 (Proton Gradient Regulation 5) has been shown to be essential for the main route of PSI cyclic electron transport, its exact function remains unclear. In transgenic Arabidopsis plants overaccumulating PGR5 in the thylakoid membrane, chloroplast development was delayed, especially in the cotyledons. Although photosynthetic electron transport was not affected during steady-state photosynthesis, a high level of non-photochemical quenching (NPQ) was transiently induced after a shift of light conditions. This phenotype was explained by elevated activity of PSI cyclic electron transport, which was monitored in an in vitro system using ruptured chloroplasts, and also in leaves. The effect of overaccumulation of PGR5 was specific to the antimycin A-sensitive pathway of PSI cyclic electron transport but not to the NAD(P)H dehydrogenase (NDH) pathway. We propose that a balanced PGR5 level is required for efficient regulation of the rate of antimycin A-sensitive PSI cyclic electron transport, although the rate of PSI cyclic electron transport is probably also regulated by other factors during steady state photosynthesis. PMID- 17913768 TI - Cortical sensory map rearrangement after spinal cord injury: fMRI responses linked to Nogo signalling. AB - Cortical sensory maps can reorganize in the adult brain in an experience dependent manner. We monitored somatosensory cortical reorganization after sensory deafferentation using functional magnetic resonance imaging (fMRI) in rats subjected to complete transection of the mid-thoracic spinal cord. Cortical representation in response to spared forelimb stimulation was observed to enlarge and invade adjacent sensory-deprived hind limb territory in the primary somatosensory cortex as early as 3 days after injury. Functional MRI also demonstrated long-term cortical plasticity accompanied by increased thalamic activation. To support the notion that alterations of cortical neuronal circuitry after spinal cord injury may underlie the fMRI changes, we quantified transcriptional activities of several genes related to cortical plasticity including the Nogo receptor (NgR), its co-receptor LINGO-1 and brain derived neurotrophic factor (BDNF), using in situ hybridization. We demonstrate that NgR and LINGO-1 are down-regulated specifically in cortical areas deprived of sensory input and in adjacent cortex from 1 day after injury, while BDNF is up-regulated. Our results demonstrate that cortical neurons react to sensory deprivation by decreasing transcriptional activities of genes encoding the Nogo receptor components in the sensory deprived and the anatomically adjacent non-deprived area. Combined with the BDNF up-regulation, these changes presumably allow structural changes in the neuropil. Our observations therefore suggest an involvement of Nogo signalling in cortical activity-dependent plasticity in the somatosensory system. In spinal cord injury, cortical reorganization as shown here can become a disadvantage, much like the situation in amblyopia or phantom sensation. Successful strategies to repair sensory pathways at the spinal cord level may not lead to proper reestablishment of cortical connections, once deprived hind limb cortical areas have been reallocated to forelimb use. In such situations, methods to control cortical plasticity, possibly by targeting Nogo signalling, may become helpful. PMID- 17913770 TI - Diversifying the options for interacting with patients. PMID- 17913771 TI - Analysing low-risk patient populations allows better discrimination between high performing and low-performing hospitals: a case study using inhospital mortality from acute myocardial infarction. AB - OBJECTIVE: To assess whether performance indicators based on administrative hospital data can be rendered more useful by stratifying them according to risk status of the patient. DESIGN: Retrospective analysis of 10 years of administrative hospital data for patients with acute myocardial infarction (AMI). Four risk groups defined by cross-classifying patient age (<75 years, 75+ years) against the presence or otherwise of at least one risk condition that predicted short-term AMI mortality. SETTING: 17 public hospitals in Queensland, Australia, with more than 50 AMI admissions annually. PARTICIPANTS: 21,537 patients admitted through the emergency department and subsequently diagnosed as having AMI. MAIN OUTCOME MEASURE: Systematic variation in standardised case fatality ratios. Systematic variation is the variation across hospitals after accounting for the Poisson variation in the number of deaths at each hospital. It was obtained from an empirical-Bayes model. Case fatality ratios were standardised according to the age, sex and risk factor profile of the patient. RESULTS: Systematic variation decreased monotonically across the four risk groups as case fatality increased (likelihood ratio test: chi(2) = 8.08, df = 1, p = 0.004). Systematic variation was largest and statistically significant (0.375; 95% CI 0.144 to 0.606) for low risk patients (<75 years with no risk conditions; case fatality rate = 2.0%) but was smallest (0.126; 0.039 to 0.212) for high-risk patients (75+ years with at least one risk condition; case fatality rate = 24.3%). CONCLUSION: Analysis of data from high-risk patients with AMI provides little opportunity to identify better-performing hospitals because there is relatively little variation across hospitals. In such patients, older age and comorbid illness are probably more important than quality of care in determining outcomes. In contrast, for low-risk patients the systematic variation was large suggesting that outcomes for such patients are more sensitive to clinical error. Analysing data for low-risk patients maximises our ability to identify best-performing hospitals and learn from their processes and structures to effect system-wide changes that will benefit all patients. PMID- 17913772 TI - Intensivist physician staffing and the process of care in academic medical centres. AB - BACKGROUND: Although intensivist physician staffing is associated with improved outcomes in critical care, little is known about the mechanism leading to this observation. OBJECTIVE: To determine the relationship between intensivist staffing and select process-based quality indicators in the intensive care unit. RESEARCH DESIGN: Retrospective cohort study in 29 academic hospitals participating in the University HealthSystem Consortium Mechanically Ventilated Patient Bundle Benchmarking Project. PATIENTS: 861 adult patients receiving prolonged mechanical ventilation in an intensive care unit. RESULTS: Patient level information on physician staffing and process-of-care quality indicators were collected on day 4 of mechanical ventilation. By day 4, 668 patients received care under a high intensity staffing model (primary intensivist care or mandatory consult) and 193 patients received care under a low intensity staffing model (optional consultation or no intensivist). Among eligible patients, those receiving care under a high intensity staffing model were more likely to receive prophylaxis for deep vein thrombosis (risk ratio 1.08, 95% CI 1.00 to 1.17), stress ulcer prophylaxis (risk ratio 1.10, 95% CI 1.03 to 1.18), a spontaneous breathing trial (risk ratio 1.37, 95% CI 0.97 to 1.94), interruption of sedation (risk ratio 1.64, 95% CI 1.13 to 2.38) and intensive insulin treatment (risk ratio 1.40, 95% CI 1.18 to 1.79) on day 4 of mechanical ventilation. Models accounting for clustering by hospital produced similar estimates of the staffing effect, except for prophylaxis against thrombosis and stress ulcers. CONCLUSIONS: High intensity physician staffing is associated with increased use of evidence based quality indicators in patients receiving mechanical ventilation. PMID- 17913773 TI - Using a Malcolm Baldrige framework to understand high-performing clinical microsystems. AB - BACKGROUND, OBJECTIVES AND METHOD: The Malcolm Baldrige National Quality Award (MBNQA) provides a set of criteria for organisational quality assessment and improvement that has been used by thousands of business, healthcare and educational organisations for more than a decade. The criteria can be used as a tool for self-evaluation, and are widely recognised as a robust framework for design and evaluation of healthcare systems. The clinical microsystem, as an organisational construct, is a systems approach for providing clinical care based on theories from organisational development, leadership and improvement. This study compared the MBNQA criteria for healthcare and the success factors of high performing clinical microsystems to (1) determine whether microsystem success characteristics cover the same range of issues addressed by the Baldrige criteria and (2) examine whether this comparison might better inform our understanding of either framework. RESULTS AND CONCLUSIONS: Both Baldrige criteria and microsystem success characteristics cover a wide range of areas crucial to high performance. Those particularly called out by this analysis are organisational leadership, work systems and service processes from a Baldrige standpoint, and leadership, performance results, process improvement, and information and information technology from the microsystem success characteristics view. Although in many cases the relationship between Baldrige criteria and microsystem success characteristics are obvious, in others the analysis points to ways in which the Baldrige criteria might be better understood and worked with by a microsystem through the design of work systems and a deep understanding of processes. Several tools are available for those who wish to engage in self-assessment based on MBNQA criteria and microsystem characteristics. PMID- 17913774 TI - Using a survey of incident reporting and learning practices to improve organisational learning at a cancer care centre. AB - OBJECTIVES: To motivate improvements in an organisational system by measuring staff perceptions of the organisation's ability to learn from incidents and by analysing their personal experience of incidents. METHODS: Respondents were questioned on the components of the incident learning system from both a personal and an organisational perspective. The respondents (n = 125) were radiotherapists, nurses, dosimetrists, doctors, and other staff at a major academic cancer centre. Responses were analysed in terms of per cent positive responses and response rate, differences between "frontline" and "support" staff, and the respondent's experience with incidents. RESULTS: Respondents were more familiar with and more positive about incident identification and reporting--the first two stages of incident learning. Their overall perception of incident learning was most influenced by the investigation and learning components of the system. Respondents in frontline positions were more positive than those in support positions about responding to, identifying and reporting incidents. Respondents reported having experienced a mean of three incidents per year, of which two were reported and two out of three of the reported incidents were investigated, and a median of two incidents being experienced and reported, but none investigated. Most incidents experienced were not captured by the organisation's existing incident reporting system. CONCLUSION: The survey tool was effective in measuring the ability of the organisation to learn from incidents. Implications of the survey results for improving organisational learning are discussed. PMID- 17913775 TI - Shared medical appointments based on the chronic care model: a quality improvement project to address the challenges of patients with diabetes with high cardiovascular risk. AB - OBJECTIVE: The epidemic proportions and management complexity of diabetes have prompted efforts to improve clinic throughput and efficiency. One method of system redesign based on the chronic care model is the Shared Medical Appointment (SMA) in which groups of patients (8-20) are seen by a multi-disciplinary team in a 1-2 h appointment. Evaluation of the impact of SMAs on quality of care has been limited. The purpose of this quality improvement project was to improve intermediate outcome measures for diabetes (A1c, SBP, LDL-cholesterol) focusing on those patients at highest cardiovascular risk. SETTING: Primary care clinic at a tertiary care academic medical center. SUBJECTS: Patients with diabetes with one or more of the following: A1c >9%, SBP blood pressure >160 mm Hg and LDL-c >130 mg/dl were targeted for potential participation; other patients were referred by their primary care providers. Patients participated in at least one SMA from 4/05 to 9/05. STUDY DESIGN: Quasi-experimental with concurrent, but non randomised controls (patients who participated in SMAs from 5/06 through 8/06; a retrospective period of observation prior to their SMA participation was used). INTERVENTION: SMA system redesign. ANALYTICAL METHODS: Paired and independent t tests, chi(2) tests and Fisher Exact tests. RESULTS: Each group had up to 8 patients. Patients participated in 1-7 visits. At the initial visit, 83.3% had A1c levels >9%, 30.6% had LDL-cholesterol levels >130 mg/dl, and 34.1% had SBP >or=160 mm Hg. Levels of A1c, LDL-c and SBP all fell significantly postintervention with a mean (95% CI) decrease of A1c 1.4 (0.8, 2.1) (p<0.001), LDL-c 14.8 (2.3, 27.4) (p = 0.022) and SBP 16.0 (9.7, 22.3) (p<0.001). There were no significant differences at baseline between control and intervention groups in terms of age, baseline intermediate outcomes, or medication use. The reductions in A1c in % and SBP were greater in the intervention group relative to the control group: 1.44 vs -0.30 (p = 0.002) for A1c and 14.83 vs 2.54 mm Hg (p = 0.04) for SBP. LDL-c reduction was also greater in the intervention group, 16.0 vs 5.37 mg/dl, but the difference was not statistically significant (p = 0.29). CONCLUSIONS: We were able to initiate a programme of group visits in which participants achieved benefits in terms of cardiovascular risk reduction. Some barriers needed to be addressed, and the operations of SMAs evolved over time. Shared medical appointments for diabetes constitute a practical system redesign that may help to improve quality of care. PMID- 17913776 TI - Monitoring quality standards in the provision of non-prescription medicines from Australian Community Pharmacies: results of a national programme. AB - INTRODUCTION: Standards of practice have been developed by the pharmacy profession to address the provision of non-prescription medicines, using a consumer-focused and risk management approach. The application of these standards in Australian community pharmacies has been monitored since 2002 by the Quality Care Pharmacy Support Centre (QCPSC). METHODS: Between September 2002 and September 2005, 7785 standards maintenance assessment visits were conducted in 4282 pharmacies, using pseudo-patient methodology. 1909 were symptom-based requests (SBRs) and 5876 were direct product-based requests (DPRs), of which 2864 were for pharmacist-only medicines (POMs) and 3012 were for pharmacy medicines (PMs). 2756 pharmacies received two visits, and 747 received three visits. A pharmacy's performance was scored out of 10 at each visit (scores 0-3: "unsatisfactory"; 4-6: "satisfactory"; and 7-10: "excellent"). RESULTS: There was wide variation in performance at baseline, with 1453 (34%) of pharmacies scoring infinity) values of the conjugated silymarin flavonolignans were 4- to 30-fold higher than those of their free fractions, with SB(B) (mean AUC(0-->infinity) = 51 and 597 microg x h/l for free and conjugated, respectively) and ISB(A) (mean AUC(0-->infinity) = 30 and 734 microg x h/l for free and conjugated, respectively) exhibiting higher AUC(0- >infinity) values in comparison with other flavonolignans. Near the plasma peak times (1-3 h), the free, sulfated, and glucuronidated flavonolignans represented approximately 17, 28, and 55% of the total silymarin, respectively. In addition, the individual silymarin flavonolignans exhibited quite different plasma profiles for both the free and conjugated fractions. These data suggest that, after oral administration, silymarin flavonolignans are quickly metabolized to their conjugates, primarily forming glucuronides, and the conjugates are primary components present in human plasma. PMID- 17913796 TI - Multidrug resistance-associated protein 2 is primarily responsible for the biliary excretion of fexofenadine in mice. AB - Previous studies implicated P-glycoprotein (P-gp) as the major transport protein responsible for the biliary excretion of fexofenadine (FEX). However, FEX biliary excretion was not impaired in P-gp- or breast cancer resistance protein (Bcrp) knockout mice or multidrug resistance-associated protein 2 (Mrp2)-deficient rats. The present study tested the hypothesis that species differences exist in the transport protein primarily responsible for FEX biliary excretion between mice and rats. Livers from Mrp2-knockout (Mrp2KO) mice and Mrp2-deficient (TR(-)) rats were perfused in a single-pass manner with 0.5 muM FEX. N-(4-[2-(1,2,3,4 Tetrahydro-6,7-dimethoxy-2-isoquinolinyl)ethyl]-phenyl)-9,10-dihydro-5-methoxy-9 oxo-4-acridine carboxamide (GF120918) (10 muM) was employed to inhibit P-gp and Bcrp. The biliary excretion rate of FEX was decreased 85% in Mrp2KO relative to wild-type mice (18.4 +/- 2.2 versus 122 +/- 34 pmol/min/g liver). In mice, more than 50% of FEX unbound intrinsic biliary clearance (CL(bile, int)(') = 3.0 ml/h/g liver) could be attributed to Mrp2 (Mrp2-dependent CL(bile, int)(') approximately 1.7 ml/h/g liver), with P-gp and Bcrp playing a minor role (P-gp- and Bcrp-dependent CL(bile, int)(') approximately 0.3 ml/h/g liver). Approximately one third of FEX CL(bile, int)(') was attributed to unidentified mechanisms in mice. In contrast to mice, FEX biliary excretion rate (245 +/- 38 and 250 +/- 25 pmol/min/g liver) and CL(bile, int)(') (9.72 +/- 2.47 and 6.49 +/- 0.68 ml/h/g liver) were comparable between TR(-) and control Wistar rats, respectively, suggesting that unidentified transport mechanism(s) can completely compensate for the loss of Mrp2 function in rats. Mrp2 clearly plays a major role in FEX biliary excretion in mice. In conclusion, remarkable species differences exist in FEX hepatobiliary transport mechanisms. PMID- 17913798 TI - Intracellular processing, glycosylation, and cell surface expression of human metapneumovirus attachment glycoprotein. AB - The biosynthesis and posttranslational processing of human metapneumovirus attachment G glycoprotein were investigated. After pulse-labeling, the G protein accumulated as three species with molecular weights of 45,000, 50,000, and 53,000 (45K, 50K, and 53K, respectively). N-Glycosidase digestion indicated that these forms represent the unglycosylated precursor and N-glycosylated intermediate products, respectively. After an appropriate chase, these three naive forms were further processed to a mature 97K form. The presence of O-linked sugars in mature G protein was confirmed by O-glycanase digestion and lectin-binding assay using Arachis hypogaea (peanut agglutinin), an O-glycan-specific lectin. In addition, in the O-glycosylation-deficient cell line (CHO ldlD cell), the G protein could not be processed to the mature form unless the exogenous Gal and GalNAc were supplemented, which provided added evidence supporting the O-linked glycosylation of G protein. The maturation of G was completely blocked by monensin but was partially sensitive to brefeldin A (BFA), suggesting the O-linked glycosylation of G initiated in the trans-Golgi compartment and terminated in the trans-Golgi network. Enzymatic deglycosylation analysis confirmed that the BFA-G was a partial mature form containing N-linked oligosaccharides and various amounts of O linked carbohydrate side chains. The expression of G protein at the cell surface could be detected by indirect immunofluorescence staining assay. Furthermore, cell surface immunoprecipitation displayed an efficient intracellular transport of G protein. PMID- 17913797 TI - Dynamics of T-cell responses and memory T cells during primary simian immunodeficiency virus infection in cynomolgus macaques. AB - Cellular immune responses make an important contribution to both the control of human immunodeficiency virus (HIV) replication and disease progression. We used a pathogenic model of SIVmac251 infection of cynomolgus macaques to longitudinally evaluate cellular immune responses in association with various rates of disease progression. We found an inverse relationship between plasma viral load and the simian immunodeficiency virus (SIV)-specific T cells responses in peripheral blood and lymph nodes. SIV-specific T-cell responses in peripheral blood were transient during primary infection, with the highest responses detected around 3 months after infection. There was also a transient increase of central memory CD8(+) T cells in peripheral blood during primary infection, and effector memory T-cell counts in peripheral lymph nodes were increased. This study emphasizes the importance of the early virus-specific immune responses in the outcome of HIV/SIV disease and provides details about the changes of virus-specific immune responses over time. PMID- 17913799 TI - Two-way antigenic cross-reactivity between severe acute respiratory syndrome coronavirus (SARS-CoV) and group 1 animal CoVs is mediated through an antigenic site in the N-terminal region of the SARS-CoV nucleoprotein. AB - In 2002, severe acute respiratory syndrome-associated coronavirus (SARS-CoV) emerged in humans, causing a global epidemic. By phylogenetic analysis, SARS-CoV is distinct from known CoVs and most closely related to group 2 CoVs. However, no antigenic cross-reactivity between SARS-CoV and known CoVs was conclusively and consistently demonstrated except for group 1 animal CoVs. We analyzed this cross reactivity by an enzyme-linked immunosorbent assay (ELISA) and Western blot analysis using specific antisera to animal CoVs and SARS-CoV and SARS patient convalescent-phase or negative sera. Moderate two-way cross-reactivity between SARS-CoV and porcine CoVs (transmissible gastroenteritis CoV [TGEV] and porcine respiratory CoV [PRCV]) was mediated through the N but not the spike protein, whereas weaker cross-reactivity occurred with feline (feline infectious peritonitis virus) and canine CoVs. Using Escherichia coli-expressed recombinant SARS-CoV N protein and fragments, the cross-reactive region was localized between amino acids (aa) 120 to 208. The N-protein fragments comprising aa 360 to 412 and aa 1 to 213 reacted specifically with SARS convalescent-phase sera but not with negative human sera in ELISA; the fragment comprising aa 1 to 213 cross-reacted with antisera to animal CoVs, whereas the fragment comprising aa 360 to 412 did not cross-react and could be a potential candidate for SARS diagnosis. Particularly noteworthy, a single substitution at aa 120 of PRCV N protein diminished the cross-reactivity. We also demonstrated that the cross-reactivity is not universal for all group 1 CoVs, because HCoV-NL63 did not cross-react with SARS-CoV. One-way cross-reactivity of HCoV-NL63 with group 1 CoVs was localized to aa 1 to 39 and at least one other antigenic site in the N-protein C terminus, differing from the cross-reactive region identified in SARS-CoV N protein. The observed cross-reactivity is not a consequence of a higher level of amino acid identity between SARS-CoV and porcine CoV nucleoproteins, because sequence comparisons indicated that SARS-CoV N protein has amino acid identity similar to that of infectious bronchitis virus N protein and shares a higher level of identity with bovine CoV N protein within the cross-reactive region. The TGEV and SARS-CoV N proteins are RNA chaperons with long disordered regions. We speculate that during natural infection, antibodies target similar short antigenic sites within the N proteins of SARS-CoV and porcine group 1 CoVs that are exposed to an immune response. Identification of the cross-reactive and non-cross-reactive N protein regions allows development of SARS-CoV-specific antibody assays for screening animal and human sera. PMID- 17913800 TI - Contribution of endocytic motifs in the cytoplasmic tail of herpes simplex virus type 1 glycoprotein B to virus replication and cell-cell fusion. AB - The use of endocytic pathways by viral glycoproteins is thought to play various functions during viral infection. We previously showed in transfection assays that herpes simplex virus type 1 (HSV-1) glycoprotein B (gB) is transported from the cell surface back to the trans-Golgi network (TGN) and that two motifs of gB cytoplasmic tail, YTQV and LL, function distinctly in this process. To investigate the role of each of these gB trafficking signals in HSV-1 infection, we constructed recombinant viruses in which each motif was rendered nonfunctional by alanine mutagenesis. In infected cells, wild-type gB was internalized from the cell surface and concentrated in the TGN. Disruption of YTQV abolished internalization of gB during infection, whereas disruption of LL induced accumulation of internalized gB in early recycling endosomes and impaired its return to the TGN. The growth of both recombinants was moderately diminished. Moreover, the fusion phenotype of cells infected with the gB recombinants differed from that of cells infected with the wild-type virus. Cells infected with the YTQV-mutated virus displayed reduced cell-cell fusion, whereas giant syncytia were observed in cells infected with the LL-mutated virus. Furthermore, blocking gB internalization or impairing gB recycling to the cell surface, using drugs or a transdominant negative form of Rab11, significantly reduced cell-cell fusion. These results favor a role for endocytosis in virus replication and suggest that gB intracellular trafficking is involved in the regulation of cell cell fusion. PMID- 17913802 TI - Antiviral antibodies are necessary to prevent cytotoxic T-lymphocyte escape in mice infected with a coronavirus. AB - Mutation within virus-derived CD8 T-cell epitopes can effectively abrogate cytotoxic T-lymphocyte (CTL) recognition and impede virus clearance in infected hosts. These so-called "CTL escape variant viruses" are commonly selected during persistent infections and are associated with rapid disease progression and increased disease severity. Herein, we tested whether antiviral antibody-mediated suppression of virus replication and subsequent virus clearance were necessary for preventing CTL escape in coronavirus-infected mice. We found that compared to wild-type mice, B-cell-deficient mice did not efficiently clear infectious virus, uniformly developed clinical disease, and harbored CTL escape variant viruses. These data directly demonstrate a critical role for antiviral antibody in protecting from the selective outgrowth of CTL escape variant viruses. PMID- 17913801 TI - Promoter- and cell-specific transcriptional transactivation by the Kaposi's sarcoma-associated herpesvirus ORF57/Mta protein. AB - The Kaposi's sarcoma-associated herpesvirus (KSHV) Mta protein, encoded by open reading frame 57, is a transactivator of gene expression that is essential for productive viral replication. Previous studies have suggested both transcriptional and posttranscriptional roles for Mta, but little is known regarding Mta's transcriptional function. In this study, we demonstrate that Mta cooperates with the KSHV lytic switch protein, Rta, to reactivate KSHV from latency, but Mta has little effect on reactivation when expressed alone. We demonstrate that the Mta and Rta proteins are expressed with similar but distinct kinetics during KSHV reactivation. In single-cell analyses, Mta expression coincides tightly with progression to full viral reactivation. We demonstrate with promoter reporter assays that while Rta activates transcription in all cell lines tested, Mta's ability to transactivate promoters, either alone or synergistically with Rta, is cell and promoter specific. In particular, Mta robustly transactivates the nut-1/PAN promoter independently of Rta in 293 and Akata-31 cells. Using nuclear run-on assays, we demonstrate that Mta stimulates transcriptional initiation in 293 cells. Rta and Mta physically interact in infected cell extracts, and this interaction requires the intact leucine repeat and central region of Rta in vitro. We demonstrate that Mta also binds to the nut 1/PAN promoter DNA in vitro and in infected cells. An Mta mutant with a lesion in a putative A/T hook domain is altered in DNA binding and debilitated in transactivation. We propose that one molecular mechanism of Mta-mediated transactivation is a direct effect on transcription by direct and indirect promoter association. PMID- 17913803 TI - Overexpression of the kaposi's sarcoma-associated herpesvirus transactivator K Rta can complement a K-bZIP deletion BACmid and yields an enhanced growth phenotype. AB - Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 (HHV8) ORF50 encodes a transactivator, K-Rta, which functions as the switch from latent to lytic virus replication. K-bZIP interacts with K-Rta and can repress its transactivation activity for some viral promoters. Both K-Rta and K-bZIP are required for origin dependent DNA replication. To determine the role of K-bZIP in the context of the viral genome, we generated a recombinant HHV8 bacterial artificial chromosome (BAC) with a deletion in the K-bZIP open reading frame. This BACmid, BAC36DeltaK8, displayed an enhanced growth phenotype with respect to virus production and accumulation of virus-encoded mRNAs measured by real-time PCR when K-Rta was used to induce the virus lytic cycle. Conversely, induction of the virus lytic cycle using tetradecanoyl phorbol acetate/n-butyrate resulted in no virus production and an aberrant gene expression pattern from BAC36DeltaK8 containing cells compared to wild-type (wt) BAC. This null virus phenotype was efficiently complemented by the expression of K-bZIP in trans, restoring virus production to wt BAC levels. Immunofluorescence staining revealed that subcellular localization of K-Rta was unchanged; however, a disruption of LANA subcellular localization was observed in cells harboring BAC36DeltaK8, suggesting that K-bZIP influences LANA localization. Coimmunoprecipitation experiments confirmed that K-bZIP interacts with LANA in BCBL-1 cells and in cotransfection assays. Lastly, the chromatin immunoprecipitation assay revealed that, in an environment where K-Rta is overexpressed and in the absence of K-bZIP, K-Rta binds to CAAT enhancer binding protein alpha sites within oriLyt, suggesting that it is K-Rta that supplies an essential replication function and that K-bZIP may serve to augment or facilitate the interaction of K-Rta with oriLyt. PMID- 17913804 TI - Tioman virus, a paramyxovirus of bat origin, causes mild disease in pigs and has a predilection for lymphoid tissues. AB - Disease manifestation, pathology, and tissue tropism following infection with Tioman virus (TioPV), a newly isolated, bat-derived paramyxovirus, was investigated in subcutaneously (n = 12) and oronasally (n = 4) inoculated pigs. Pigs were either asymptomatic or developed pyrexia, but all of the animals produced neutralizing antibodies. The virus (viral antigen and/or genome) was detected in lymphocytes of the thymus, tonsils, spleen, lymph nodes and Peyer's patches (ileum), tonsillar epithelium, and thymic epithelioreticular cells. Virus was isolated from oral swabs but not from urine. Our findings suggest that the pig could act as an intermediate or amplifying host for TioPV and that oral secretion is a possible means of viral transmission. PMID- 17913805 TI - Human herpesvirus 6A (HHV-6A) and HHV-6B alter E2F1/Rb pathways and E2F1 localization and cause cell cycle arrest in infected T cells. AB - E2F transcription factors play pivotal roles in controlling the expression of genes involved in cell viability as well as genes involved in cell death. E2F1 is an important constituent of this protein family, which thus far contains eight members. The interaction of E2F1 with its major regulator, retinoblastoma protein (Rb), has been studied extensively in the past two decades, concentrating on the role of E2F1 in transcriptional regulation and the role of Rb in cell replication and cancer formation. Additionally, the effect of viral infections on E2F1/Rb interactions has been analyzed for different viruses, concentrating on cell division, which is essential for viral replication. In the present study, we monitored E2F1-Rb interactions during human herpesvirus 6A (HHV-6A) and HHV-6B infections of SupT1 T cells. The results have shown the following dramatic alterations in E2F1-Rb pathways compared to the pathways of parallel mock infected control cultures. (i) The E2F1 levels were elevated during viral infections. (ii) The cellular localization of E2F1 was dramatically altered, and it was found to accumulate both in the cytoplasmic and nuclear fractions, as opposed to the strict nuclear localization seen in the mock-infected cells. (iii) Although E2F1 expression was elevated, two exemplary target genes, cyclin E and MCM5, were not upregulated. (iv) The Rb protein was dephosphorylated early postinfection, a trait that also occurred with UV-inactivated virus. (v) Infection was associated with significant reduction of E2F1/Rb complexing. (vi) HHV-6 infections were accompanied by cell cycle arrest. The altered E2F1-Rb interactions and functions might contribute to the observed cell cycle arrest. PMID- 17913806 TI - Mapping protease inhibitor resistance to human immunodeficiency virus type 1 sequence polymorphisms within patients. AB - Resistance genotyping provides an important resource for the clinical management of patients infected with human immunodeficiency virus type 1 (HIV-1). However, resistance to protease (PR) inhibitors (PIs) is a complex phenotype shaped by interactions among nearly half of the residues in HIV-1 PR. Previous studies of the genetic basis of PI resistance focused on fixed substitutions among populations of HIV-1, i.e., host-specific adaptations. Consequently, they are susceptible to a high false discovery rate due to founder effects. Here, we employ sequencing "mixtures" (i.e., ambiguous base calls) as a site-specific marker of genetic variation within patients that is independent of the phylogeny. We demonstrate that the transient response to selection by PIs is manifested as an excess of nonsynonymous mixtures. Using a sample of 5,651 PR sequences isolated from both PI-naive and -treated patients, we analyze the joint distribution of mixtures and eight PIs as a Bayesian network, which distinguishes residue-residue interactions from direct associations with PIs. We find that selection for resistance is associated with the emergence of nonsynonymous mixtures in two distinct groups of codon sites clustered along the substrate cleft and distal regions of PR, respectively. Within-patient evolution at several positions is independent of PIs, including those formerly postulated to be involved in resistance. These positions are under strong positive selection in the PI-naive patient population, implying that other factors can produce spurious associations with resistance, e.g., mutational escape from the immune response. PMID- 17913807 TI - Adhesion molecule interactions facilitate human immunodeficiency virus type 1 induced virological synapse formation between T cells. AB - Human immunodeficiency virus type 1 (HIV-1) can spread between CD4+ T cells by using a virological synapse (VS). The VS assembly is a cytoskeleton-driven process dependent on HIV-1 envelope glycoprotein (Env)-receptor engagement and is hypothesized to require adhesion molecule interactions. Here we demonstrate that leukocyte function-associated antigen 1 (LFA-1), intercellular adhesion molecule 1 (ICAM-1), and ICAM-3 are enriched at the VS and that inhibition of these interactions influences conjugate formation and reduces VS assembly. Moreover, CD4+ T cells deficient in LFA-1 or with modified LFA-1 function were less able to support VS assembly and cell-cell transfer of HIV-1. Thus, cognate adhesion molecule interactions at the VS are important for HIV-1 spread between T cells. PMID- 17913808 TI - Ubiquitin depletion and dominant-negative VPS4 inhibit rhabdovirus budding without affecting alphavirus budding. AB - The budding reactions of a number of enveloped viruses use the cellular machinery involved in the formation of the luminal vesicles of endosomal multivesicular bodies (MVB). Budding of these viruses is dependent on the presence of specific late-domain motifs in membrane-associated viral proteins. Such budding reactions usually involve ubiquitin and are blocked by expression of an ATPase-deficient form of VPS4, a cellular AAA+ ATPase believed to be required late in the MVB pathway for the disassembly/release of the MVB machinery. Here we examined the role of the MVB pathway in the budding of the late-domain-containing rhabdovirus vesicular stomatitis virus (VSV) and the alphavirus Semliki Forest virus (SFV). We tested early and late steps in the MVB pathway by depleting ubiquitin with the proteasome inhibitor MG-132 and by using cell lines inducibly expressing VPS4A or VPS4B protein. As previously shown, VSV budding was strongly dependent on ubiquitin. In contrast to the findings of previous studies with VPS4A, expression of ATPase-deficient mutants of either VPS4A or VPS4B inhibited VSV budding. Inhibition by VPS4 required the presence of the PPPY late domain on the VSV matrix protein and resulted in the accumulation of nonreleased VSV particles at the plasma membrane. In contrast, SFV budding was independent of both ubiquitin and the activity of VPS4, perhaps reflecting the important role of the highly organized envelope protein lattice during alphavirus budding. PMID- 17913809 TI - Human seminal plasma abrogates the capture and transmission of human immunodeficiency virus type 1 to CD4+ T cells mediated by DC-SIGN. AB - Dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) is expressed by dendritic cells (DCs) at mucosal surfaces and appears to play an important role in the dissemination of human immunodeficiency virus type 1 (HIV-1) infection. DC-SIGN binds HIV-1 gp120 and efficiently transmits the virus to T CD4(+) cells, which become the center of viral replication. Semen represents the main vector for HIV-1 dissemination worldwide. In the present study we show that human seminal plasma (SP), even when used at very high dilutions (1:10(4) to 1:10(5)), markedly inhibits the capture and transmission of HIV-1 to T CD4(+) cells mediated by both DCs and B-THP-1-DC-SIGN cells. In contrast, SP does not inhibit the capture of HIV-1 by DC-SIGN-negative target cells, such as the T-cell line SupT-1, monocytes, and activated peripheral blood mononuclear cells. The SP inhibitor has a high molecular mass (>100 kDa) and directly interacts with DC-SIGN-positive target cells but not with HIV-1. Moreover, the inhibitor binds to concanavalin A, suggesting that it contains high mannose N-linked carbohydrates. Of note, using biotin-labeled SP we found that the binding of SP components to DCs was abrogated by mannan, while their interaction with B-THP-1 cells was almost completely dependent on the expression of DC-SIGN. Since epithelium integrity is often compromised after vaginal or anal intercourse, as well as in the presence of ulcerative-sexually transmitted diseases, our results support the notion that components of the SP might be able to access to the subepithelium, inhibiting the recognition of HIV-1 gp120 by DC SIGN-positive DCs. PMID- 17913810 TI - Chaperones activate hepadnavirus reverse transcriptase by transiently exposing a C-proximal region in the terminal protein domain that contributes to epsilon RNA binding. AB - All hepatitis B viruses replicate by protein-primed reverse transcription, employing a specialized reverse transcriptase, P protein, that carries a unique terminal protein (TP) domain. To initiate reverse transcription, P protein must bind to a stem-loop, epsilon, on the pregenomic RNA template. TP then provides a Y residue for covalent attachment of the first nucleotide of an epsilon-templated DNA oligonucleotide (priming reaction) that serves to initiate full-length minus strand DNA synthesis. epsilon binding requires the chaperone-dependent conversion of inactive P protein into an activated, metastable form designated P*. However, how P* differs structurally from P protein is not known. Here we used an in vitro reconstitution system for active duck hepatitis B virus P combined with limited proteolysis, site-specific antibodies, and defined P mutants to structurally compare nonactivated versus chaperone-activated versus primed P protein. The data show that Hsp70 action, under conditions identical to those required for functional activation, transiently exposes the C proximal TP region which is, probably directly, involved in epsilon RNA binding. Notably, after priming and epsilon RNA removal, a very similar new conformation appears stable without further chaperone activity; hence, the activation of P protein is triggered by energy-consuming chaperone action but may be completed by template RNA binding. PMID- 17913811 TI - ERK- and JNK-dependent signaling pathways contribute to Bombyx mori nucleopolyhedrovirus infection. AB - Mitogen-activated protein kinases (MAPKs) often play important roles in virus infection. To explore intracellular signaling pathways induced by baculovirus infection, we examined the involvement of MAPKs in Bombyx mori nucleopolyhedrovirus (BmNPV) infection of BmN cells. We found that specific inhibitors of extracellular signal-regulated kinase (ERK) kinase and c-Jun NH(2) terminal kinase (JNK) significantly reduced occlusion body (OB) formation and budded virus (BV) production. Next, we quantified OB and BV production after applying the inhibitors at different times postinfection (p.i.). The inhibitors significantly reduced OB and BV production to various extents when applied at 12 h p.i., indicating that the reduction of BmNPV infectivity by these inhibitors occurs at the late stage of infection. Also, we observed that these inhibitors markedly repressed or deregulated the expression of delayed early, late, and very late gene products. Western blot analysis using phospho-MAPK-specific antibodies showed that ERK and JNK were activated at the late stage of BmNPV infection. In addition, the magnitude and pattern of MAPK activation were dependent on the multiplicity of infection. To verify the effects of the inhibitors on BmNPV infection, we also attempted to knock down the B. mori genes BmErk and BmJnk, which encode ERK and JNK, respectively. Knockdown of BmErk and BmJnk resulted in the reduced production of OBs and BVs, confirming that BmERK and BmJNK are involved in the BmNPV infection process. Taken together, these results indicate that the activation of MAPK signaling pathways is required for efficient infection by BmNPV. PMID- 17913812 TI - Prion strain- and species-dependent effects of antiprion molecules in primary neuronal cultures. AB - Transmissible spongiform encephalopathies (TSE) arise as a consequence of infection of the central nervous system by prions and are incurable. To date, most antiprion compounds identified by in vitro screening failed to exhibit therapeutic activity in animals, thus calling for new assays that could more accurately predict their in vivo potency. Primary nerve cell cultures are routinely used to assess neurotoxicity of chemical compounds. Here, we report that prion strains from different species can propagate in primary neuronal cultures derived from transgenic mouse lines overexpressing ovine, murine, hamster, or human prion protein. Using this newly developed cell system, the activity of three generic compounds known to cure prion-infected cell lines was evaluated. We show that the antiprion activity observed in neuronal cultures is species or strain dependent and recapitulates to some extent the activity reported in vivo in rodent models. Therefore, infected primary neuronal cultures may be a relevant system in which to investigate the efficacy and mode of action of antiprion drugs, including toward human transmissible spongiform encephalopathy agents. PMID- 17913813 TI - Temperature-sensitive mutations in the putative herpes simplex virus type 1 terminase subunits pUL15 and pUL33 preclude viral DNA cleavage/packaging and interaction with pUL28 at the nonpermissive temperature. AB - Terminases comprise essential components of molecular motors required to package viral DNA into capsids in a variety of DNA virus systems. Previous studies indicated that the herpes simplex virus type 1 U(L)15 protein (pU(L)15) interacts with the pU(L)28 moiety of a pU(L)28-pU(L)33 complex to form the likely viral terminase. In the current study, a novel temperature-sensitive mutant virus was shown to contain a mutation in U(L)33 codon 61 predicted to change threonine to proline. At the nonpermissive temperature, this virus, designated ts8-22, replicated viral DNA and produced capsids that became enveloped at the inner nuclear membrane but failed to form plaques or to cleave or package viral DNA. Incubation at the nonpermissive temperature also precluded coimmunoprecipitation of U(L)33 protein with its normal interaction partners encoded by U(L)28 and U(L)15 in ts8-22-infected cells and with pU(L)28 in transient-expression assays. Moreover, a temperature-sensitive mutation in U(L)15 precluded coimmunoprecipitation of pU(L)15 with the U(L)28 and U(L)33 proteins at the nonpermissive temperature. We conclude that interactions between putative terminase components are tightly linked to successful viral DNA cleavage and packaging. PMID- 17913814 TI - Allogeneic differences in the dependence on CD4+ T-cell help for virus-specific CD8+ T-cell differentiation. AB - CD4(+) T-cell help enables antiviral CD8(+) T cells to differentiate into fully competent memory cells and sustains CD8(+) T-cell-mediated immunity during persistent virus infection. We recently reported that mice of C57BL/6 and C3H strains differ in their dependence on CD28 and CD40L costimulation for long-term control of infection by polyoma virus, a persistent mouse pathogen. In this study, we asked whether mice of these inbred strains also vary in their requirement for CD4(+) T-cell help for generating and maintaining polyoma virus specific CD8(+) T cells. CD4(+) T-cell-depleted C57BL/6 mice mounted a robust antiviral CD8(+) T-cell response during acute infection, whereas unhelped CD8(+) T-cell effectors in C3H mice were functionally impaired during acute infection and failed to expand upon antigenic challenge during persistent infection. Using (C57BL/6 x C3H)F(1) mice, we found that the dispensability for CD4(+) T-cell help for the H-2(b)-restricted polyoma virus-specific CD8(+) T-cell response during acute infection extends to the H-2(k)-restricted antiviral CD8(+) T cells. Our findings demonstrate that dependence on CD4(+) T-cell help for antiviral CD8(+) T cell effector differentiation can vary among allogeneic strains of inbred mice. PMID- 17913815 TI - Interaction of vesicular stomatitis virus P and N proteins: identification of two overlapping domains at the N terminus of P that are involved in N0-P complex formation and encapsidation of viral genome RNA. AB - The nucleocapsid (N) protein of nonsegmented negative-strand (NNS) RNA viruses, when expressed in eukaryotic cells, aggregates and forms nucleocapsid-like complexes with cellular RNAs. The phosphoprotein (P) has been shown to prevent such aggregation by forming a soluble complex with the N protein free from cellular RNAs (designated N(0)). The N(0)-P complex presumably mediates specific encapsidation of the viral genome RNA. The precise mechanism by which the P protein carries out this function remains unclear. Here, by using a series of deleted and truncated mutant forms of the P protein of vesicular stomatitis virus (VSV), Indiana serotype, we present evidence that the N-terminal 11 to 30 amino acids (aa) of the P protein are essential in keeping the N protein soluble. Furthermore, glutathione S-transferase fused to the N-terminal 40 aa by itself is able to form the N(0)-P complex. Interestingly, the N-terminal 40-aa stretch failed to interact with the viral genome N-RNA template whereas the C-terminal 72 aa of the P protein interacted specifically with the latter. With an in vivo VSV minigenome transcription system, we further show that a deletion mutant form of P (PDelta1-10) lacking the N-terminal 10 aa which is capable of forming the N(0)-P complex was unable to support VSV minigenome transcription, although it efficiently supported transcription in vitro in a transcription-reconstitution reaction when used as purified protein. However, the same mutant protein complemented minigenome transcription when expressed together with a transcription-defective P deletion mutant protein containing N-terminal aa 1 to 210 (PDeltaII+III). Since the minigenome RNA needs to be encapsidated before transcription ensues, it seems that the entire N-terminal 210 aa are required for efficient genome RNA encapsidation. Taking these results together, we conclude that the N-terminal 11 to 30 aa are required for N(0)-P complex formation but the N-terminal 210 aa are required for genome RNA encapsidation. PMID- 17913816 TI - NSm protein of Rift Valley fever virus suppresses virus-induced apoptosis. AB - Rift Valley fever virus (RVFV) is a member of the genus Phlebovirus within the family Bunyaviridae. It can cause severe epidemics among ruminants and fever, myalgia, a hemorrhagic syndrome, and/or encephalitis in humans. The RVFV M segment encodes the NSm and 78-kDa proteins and two major envelope proteins, Gn and Gc. The biological functions of the NSm and 78-kDa proteins are unknown; both proteins are dispensable for viral replication in cell cultures. To determine the biological functions of the NSm and 78-kDa proteins, we generated the mutant virus arMP-12-del21/384, carrying a large deletion in the pre-Gn region of the M segment. Neither NSm nor the 78-kDa protein was synthesized in arMP-12-del21/384 infected cells. Although arMP-12-del21/384 and its parental virus, arMP-12, showed similar growth kinetics and viral RNA and protein accumulation in infected cells, arMP-12-del21/384-infected cells induced extensive cell death and produced larger plaques than did arMP-12-infected cells. arMP-12-del21/384 replication triggered apoptosis, including the cleavage of caspase-3, the cleavage of its downstream substrate, poly(ADP-ribose) polymerase, and activation of the initiator caspases, caspase-8 and -9, earlier in infection than arMP-12. NSm expression in arMP-12-del21/384-infected cells suppressed the severity of caspase 3 activation. Further, NSm protein expression inhibited the staurosporine-induced activation of caspase-8 and -9, demonstrating that other viral proteins were dispensable for NSm's function in inhibiting apoptosis. RVFV NSm protein is the first identified Phlebovirus protein that has an antiapoptotic function. PMID- 17913817 TI - Increased immunogenicity of a DNA-launched Venezuelan equine encephalitis virus based replicon DNA vaccine. AB - A novel genetic vaccine that is based on a Venezuelan equine encephalitis virus (VEE) replicon launched from plasmid DNA is described. The plasmid encodes a VEE replicon under the transcriptional control of the cytomegalovirus immediate-early promoter (VEE DNA). The VEE DNA consistently expressed 3- to 15-fold more green fluorescent protein in vitro than did a conventional DNA vaccine. Furthermore, transfection with the DNA-launched VEE replicon induced apoptosis and type I interferon production. Inoculation of mice with VEE DNA encoding human immunodeficiency virus type 1 gp160 significantly increased humoral responses by several orders of magnitude compared to an equal dose of a conventional DNA vaccine. These increases were also observed at 10- and 100-fold-lower doses of the VEE DNA. Cellular immune responses measured by gamma interferon and interleukin 2 enzyme-linked immunospot assay were significantly higher in mice immunized with the VEE DNA at decreased doses. The immune responses induced by the VEE DNA-encoded antigen, however, were independent of an intact type I interferon signaling pathway. Moreover, the DNA-launched VEE replicon induced an efficient prime to a VEE replicon particle (VRP) boost, increasing humoral and cellular immunity by at least 1 order of magnitude compared to VEE DNA only. Importantly, immunization with VEE DNA, as opposed to VRP, did not induce any anti-VRP neutralizing antibodies. Increased potency of DNA vaccines and reduced vector immunity may ultimately have an impact on the design of vaccination strategies in humans. PMID- 17913818 TI - Identification of regions and residues in feline junctional adhesion molecule required for feline calicivirus binding and infection. AB - The feline junctional adhesion molecule A (fJAM-A) is a functional receptor for feline calicivirus (FCV). fJAM-A is a member of the immunoglobulin superfamily (IgSF) and consists of two Ig-like extracellular domains (D1 and D2), a membrane spanning domain, and a short cytoplasmic tail. To identify regions of fJAM-A that interact with FCV, we purified recombinant fJAM-A ectodomain and D1 and D2 domains. We found that preincubation of FCV with the ectodomain or D1 was sufficient to inhibit FCV infection in plaque reduction assays. In enzyme-linked immunosorbent assays, FCV binding to fJAM-A ectodomain was concentration dependent and saturable; however, FCV bound D1 alone weakly and was unable to bind D2. To characterize FCV binding to surface-expressed fJAM-A, we transfected truncated and chimeric forms of fJAM-A into a nonpermissive cell line and assayed binding by flow cytometry. Only D1 was necessary for FCV binding to cells; all other domains could be replaced. Using a structure-guided mutational approach, we identified three mutants of fJAM-A within D1 (D42N, K43N, and S97A) that exhibited significantly decreased capacities to bind FCV. In contrast to our finding that D1 mediated FCV binding, we found that all domains of fJAM-A were necessary to confer susceptibility to FCV infection. Furthermore, surface expression of fJAM-A was not sufficient to permit FCV infection by all of the isolates we investigated. This indicates that (i) other cellular factors are required to permit productive FCV infection and (ii) individual FCV isolates differ in the factors they require. PMID- 17913819 TI - Analysis of Venezuelan equine encephalitis virus capsid protein function in the inhibition of cellular transcription. AB - The encephalitogenic New World alphaviruses, including Venezuelan (VEEV), eastern (EEEV), and western equine encephalitis viruses, constitute a continuing public health threat in the United States. They circulate in Central, South, and North America and have the ability to cause fatal disease in humans and in horses and other domestic animals. We recently demonstrated that these viruses have developed the ability to interfere with cellular transcription and use it as a means of downregulating a cellular antiviral response. The results of the present study suggest that the N-terminal, approximately 35-amino-acid-long peptide of VEEV and EEEV capsid proteins plays the most critical role in the downregulation of cellular transcription and development of a cytopathic effect. The identified VEEV-specific peptide C(VEE)33-68 includes two domains with distinct functions: the alpha-helix domain, helix I, which is critically involved in supporting the balance between the presence of the protein in the cytoplasm and nucleus, and the downstream peptide, which might contain a functional nuclear localization signal(s). The integrity of both domains not only determines the intracellular distribution of the VEEV capsid but is also essential for direct capsid protein functioning in the inhibition of transcription. Our results suggest that the VEEV capsid protein interacts with the nuclear pore complex, and this interaction correlates with the protein's ability to cause transcriptional shutoff and, ultimately, cell death. The replacement of the N-terminal fragment of the VEEV capsid by its Sindbis virus-specific counterpart in the VEEV TC-83 genome does not affect virus replication in vitro but reduces cytopathogenicity and results in attenuation in vivo. These findings can be used in designing a new generation of live, attenuated, recombinant vaccines against the New World alphaviruses. PMID- 17913820 TI - Type I interferon production during herpes simplex virus infection is controlled by cell-type-specific viral recognition through Toll-like receptor 9, the mitochondrial antiviral signaling protein pathway, and novel recognition systems. AB - Recognition of viruses by germ line-encoded pattern recognition receptors of the innate immune system is essential for rapid production of type I interferon (IFN) and early antiviral defense. We investigated the mechanisms of viral recognition governing production of type I IFN during herpes simplex virus (HSV) infection. We show that early production of IFN in vivo is mediated through Toll-like receptor 9 (TLR9) and plasmacytoid dendritic cells, whereas the subsequent alpha/beta IFN (IFN-alpha/beta) response is derived from several cell types and induced independently of TLR9. In conventional DCs, the IFN response occurred independently of viral replication but was dependent on viral entry. Moreover, using a HSV-1 UL15 mutant, which fails to package viral DNA into the virion, we found that entry-dependent IFN induction also required the presence of viral genomic DNA. In macrophages and fibroblasts, where the virus was able to replicate, HSV-induced IFN-alpha/beta production was dependent on both viral entry and replication, and ablated in cells unable to signal through the mitochondrial antiviral signaling protein pathway. Thus, during an HSV infection in vivo, multiple mechanisms of pathogen recognition are active, which operate in cell-type- and time-dependent manners to trigger expression of type I IFN and coordinate the antiviral response. PMID- 17913821 TI - Dendritic cells are less susceptible to human immunodeficiency virus type 2 (HIV 2) infection than to HIV-1 infection. AB - Human immunodeficiency virus type 1 (HIV-1) infection of dendritic cells (DCs) has been documented in vivo and may be an important contributor to HIV-1 transmission and pathogenesis. HIV-1-specific CD4(+) T cells respond to HIV antigens presented by HIV-1-infected DCs and in this process become infected, thereby providing a mechanism through which HIV-1-specific CD4(+) T cells could become preferentially infected in vivo. HIV-2 disease is attenuated with respect to HIV-1 disease, and host immune responses are thought to be contributory. Here we investigated the susceptibility of primary myeloid DCs (mDCs) and plasmacytoid DCs (pDCs) to infection by HIV-2. We found that neither CCR5-tropic primary HIV-2 isolates nor a lab-adapted CXCR4-tropic HIV-2 strain could efficiently infect mDCs or pDCs, though these viruses could infect primary CD4(+) T cells in vitro. HIV-2-exposed mDCs were also incapable of transferring virus to autologous CD4(+) T cells. Despite this, we found that HIV-2-specific CD4(+) T cells contained more viral DNA than memory CD4(+) T cells of other specificities in vivo. These data suggest that either infection of DCs is not an important contributor to infection of HIV-2-specific CD4(+) T cells in vivo or that infection of DCs by HIV-2 occurs at a level that is undetectable in vitro. The frequent carriage of HIV-2 DNA within HIV-2-specific CD4(+) T cells, however, does not appear to be incompatible with preserved numbers and functionality of HIV-2-specific CD4(+) T cells in vivo, suggesting that additional mechanisms contribute to maintenance of HIV-2 specific CD4(+) T-cell help in vivo. PMID- 17913822 TI - A self-excisable infectious bacterial artificial chromosome clone of varicella zoster virus allows analysis of the essential tegument protein encoded by ORF9. AB - In order to facilitate the generation of mutant viruses of varicella-zoster virus (VZV), the agent causing varicella (chicken pox) and herpes zoster (shingles), we generated a full-length infectious bacterial artificial chromosome (BAC) clone of the P-Oka strain. First, mini-F sequences were inserted into a preexisting VZV cosmid, and the SuperCos replicon was removed. Subsequently, mini-F-containing recombinant virus was generated from overlapping cosmid clones, and full-length VZV DNA recovered from the recombinant virus was established in Escherichia coli as an infectious BAC. An inverted duplication of VZV genomic sequences within the mini-F replicon resulted in markerless excision of vector sequences upon virus reconstitution in eukaryotic cells. Using the novel tool, the role in VZV replication of the major tegument protein encoded by ORF9 was investigated. A markerless point mutation introduced in the start codon by two-step en passant Red mutagenesis abrogated ORF9 expression and resulted in a dramatic growth defect that was not observed in a revertant virus. The essential nature of ORF9 for VZV replication was ultimately confirmed by restoration of the growth of the ORF9-deficient mutant virus using trans-complementation via baculovirus-mediated gene transfer. PMID- 17913823 TI - Differential activation of human monocyte-derived and plasmacytoid dendritic cells by West Nile virus generated in different host cells. AB - Dendritic cells (DCs) play a central role in innate immunity and antiviral responses. In this study, we investigated the production of alpha interferon (IFN alpha) and inducible chemokines by human monocyte-derived dendritic cells (mDCs) and plasmacytoid dendritic cells (pDCs) infected with West Nile virus (WNV), an emergent pathogen whose infection can lead to severe cases of encephalitis in the elderly, children, and immunocompromised individuals. Our experiments demonstrated that WNV grown in mammalian cells (WNV(Vero)) was a potent inducer of IFN-alpha secretion in pDCs and, to a lesser degree, in mDCs. The ability of WNV(Vero) to induce IFN-alpha in pDCs did not require viral replication and was prevented by the treatment of cells with bafilomycin A1 and chloroquine, suggesting that it was dependent on endosomal Toll-like receptor recognition. On the other hand, IFN-alpha production in mDCs required viral replication and was associated with the nuclear translocation of IRF3 and viral antigen expression. Strikingly, pDCs failed to produce IFN-alpha when stimulated with WNV grown in mosquito cells (WNV(C7/10)), while mDCs responded similarly to WNV(Vero) or WNV(C7/10). Moreover, the IFN-dependent chemokine IP-10 was produced in substantial amounts by pDCs in response to WNV(Vero) but not WNV(C7/10), while interleukin-8 was produced in greater amounts by mDCs infected with WNV(C7/10) than in those infected with WNV(Vero). These findings suggest that cell-specific mechanisms of WNV recognition leading to the production of type I IFN and inflammatory chemokines by DCs may contribute to both the innate immune response and disease pathogenesis in human infections. PMID- 17913825 TI - Human apolipoprotein e is required for infectivity and production of hepatitis C virus in cell culture. AB - Recent advances in reverse genetics of hepatitis C virus (HCV) made it possible to determine the properties and biochemical compositions of HCV virions. Sedimentation analysis and characterization of HCV RNA-containing particles produced in the cultured cells revealed that HCV virions cover a large range of heterogeneous densities in sucrose gradient. The fractions of low densities are infectious, while the higher-density fractions containing the majority of HCV virion RNA are not. HCV core protein and apolipoprotein B and apolipoprotein E (apoE) were detected in the infectious HCV virions. The level of apoE correlates very well with HCV infectivity. Both apoE- and HCV E2-specific monoclonal antibodies precipitated HCV, demonstrating that HCV virions contain apoE and E2 proteins. apoE-specific monoclonal antibodies efficiently neutralized HCV infectivity in a dose-dependent manner, resulting in a reduction of infectious HCV by nearly 4 orders of magnitude. The knockdown of apoE expression by specific small interfering RNAs (siRNAs) remarkably reduced the levels of intracellular as well as secreted HCV virions. The apoE siRNA suppressed HCV production by more than 100-fold at 50 nM. These findings demonstrate that apoE is required for HCV virion infectivity and production, suggesting that HCV virions are assembled as apoE-enriched lipoprotein particles. Our findings also identified apoE as a novel target for discovery and development of antiviral drugs and monoclonal antibodies to suppress HCV virion formation and infection. PMID- 17913824 TI - Targeted deletion of regions rich in immune-evasive genes from the cytomegalovirus genome as a novel vaccine strategy. AB - Human cytomegalovirus (CMV), a ubiquitous human pathogen, is a leading cause of congenital infections and represents a serious health risk for the immunosuppressed patient. A vaccine against CMV is currently not available. CMV is characterized by its large genome and by multiple genes modulating the immunity of the host, which cluster predominantly at genome termini. Here, we tested whether the deletion of gene blocks rich in immunomodulatory genes could be used as a novel concept in the generation of immunogenic but avirulent, herpesvirus vaccines. To generate an experimental CMV vaccine, we selectively deleted 32 genes from the mouse cytomegalovirus (MCMV) genome. The resulting mutant grew to titers similar to that of wild-type MCMV in vitro. In vivo, the mutant was 10,000-fold attenuated and well tolerated, even by highly susceptible mice deficient for B, T, and NK cells or for the interferon type I receptor. Equally relevant for safety concerns, immune suppression did not lead to the mutant's reactivation from latency. Immunization with the replication-competent mutant, but not with inactivated virus, resulted in protective immunity, which increased over time. Vaccination induced MCMV-specific antibodies and a strong T cell response. We propose that a targeted and rational approach can improve future herpesvirus vaccines and vaccine vectors. PMID- 17913826 TI - RNA binding domain of Jamestown Canyon virus S segment RNAs. AB - Jamestown Canyon virus (JCV) is a member of the Bunyaviridae family, Orthobunyavirus genus, California serogroup. Replication and, ultimately, assembly and packaging rely on the process of encapsidation. Therefore, the ability of viral RNAs (vRNAs) (genomic and antigenomic) to interact with the nucleocapsid protein (N protein) and the location of this binding domain on the RNAs are of interest. The questions to be addressed are the following. Where is the binding domain located on both the vRNA and cRNA strands, is this RNA bound when double or single stranded, and does this identified region have the ability to transform the binding potential of nonviral RNA? Full-length viral and complementary S segment RNA, as well as 3' deletion mutants of both vRNA and cRNA, nonviral RNA, and hybrid viral/nonviral RNA, were analyzed for their ability to interact with bacterially expressed JCV N protein. RNA-nucleocapsid interactions were examined by UV cross-linking, filter binding assays, and the generation of hybrid RNA to help define the area responsible for RNA-protein binding. The assays identified the region responsible for binding to the nucleocapsid as being contained within the 5' half of both the genomic and antigenomic RNAs. This region, if placed within nonviral RNA, is capable of altering the binding potential of nonviral RNA to levels seen with wild-type vRNAs. PMID- 17913827 TI - TRAIL is a novel antiviral protein against dengue virus. AB - Dengue fever is an important tropical illness for which there is currently no virus-specific treatment. To shed light on mechanisms involved in the cellular response to dengue virus (DV), we assessed gene expression changes, using Affymetrix GeneChips (HG-U133A), of infected primary human cells and identified changes common to all cells. The common response genes included a set of 23 genes significantly induced upon DV infection of human umbilical vein endothelial cells (HUVECs), dendritic cells (DCs), monocytes, and B cells (analysis of variance, P < 0.05). Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), one of the common response genes, was identified as a key link between type I and type II interferon response genes. We found that DV induces TRAIL expression in immune cells and HUVECs at the mRNA and protein levels. The induction of TRAIL expression by DV was found to be dependent on an intact type I interferon signaling pathway. A significant increase in DV RNA accumulation was observed in anti-TRAIL antibody-treated monocytes, B cells, and HUVECs, and, conversely, a decrease in DV RNA was seen in recombinant TRAIL-treated monocytes. Furthermore, recombinant TRAIL inhibited DV titers in DV-infected DCs by an apoptosis independent mechanism. These data suggest that TRAIL plays an important role in the antiviral response to DV infection and is a candidate for antiviral interventions against DV. PMID- 17913829 TI - Human papillomavirus type 16 E7 oncoprotein associates with the centrosomal component gamma-tubulin. AB - Expression of a high-risk human papillomavirus (HPV) E7 oncoprotein is sufficient to induce aberrant centrosome duplication in primary human cells. The resulting centrosome-associated mitotic abnormalities have been linked to the development of aneuploidy. HPV type 16 (HPV16) E7 induces supernumerary centrosomes through a mechanism that is at least in part independent of the inactivation of the retinoblastoma tumor suppressor pRb and is dependent on cyclin-dependent kinase 2 activity. Here, we show that HPV16 E7 can concentrate around mitotic spindle poles and that a small pool of HPV16 E7 is associated with centrosome fractions isolated by sucrose density gradient centrifugation. The targeting of HPV16 E7 to the centrosome, however, was not sufficient for centrosome overduplication. Nonetheless, we found that HPV16 E7 can associate with the centrosomal regulator gamma-tubulin and that the recruitment of gamma-tubulin to the centrosome is altered in HPV16 E7-expressing cells. Since the association of HPV16 E7 with gamma-tubulin is independent of pRb, p107, and p130, our results suggest that the association with gamma-tubulin contributes to the pRb/p107/p130-independent ability of HPV16 E7 to subvert centrosome homeostasis. PMID- 17913828 TI - RNA editing of the human herpesvirus 8 kaposin transcript eliminates its transforming activity and is induced during lytic replication. AB - Human herpesvirus 8 is the etiologic agent associated with Kaposi's sarcoma and primary effusion lymphoma (PEL). The K12 RNA, which produces as many as three variants of the kaposin protein, as well as a microRNA, is the most abundant transcript expressed in latent Kaposi's sarcoma-associated herpesvirus infection, and yet it is also induced during lytic replication. The portion of the transcript that includes the microRNA and the kaposin A sequence has been shown to have tumorigenic potential. Genome coordinate 117990, which is within this transcript, has been found to be heterogeneous, primarily in RNAs but also among viral DNA sequences. This sequence heterogeneity affects an amino acid in kaposins A and C and the microRNA. The functional effects of this sequence heterogeneity have not been studied, and its origin has not been definitively settled; both RNA editing and heterogeneity at the level of the viral genome have been proposed. Here, we show that transcripts containing A at position 117990 are tumorigenic, while those with G at this position are not. Using a highly sensitive quantitative assay, we observed that, in PEL cells under conditions where more than 60% of cDNAs derived from K12 RNA transcripts have G at coordinate 117990, there is no detectable G in the viral DNA sequence at this position, only A. This result is consistent with RNA editing by one of the host RNA adenosine deaminases (ADARs). Indeed, we observed that purified human ADAR1 efficiently edits K12 RNA in vitro. Remarkably, the amount of editing correlated with the replicative state of the virus; editing levels were nearly 10-fold higher in cells treated to induce lytic viral replication. These results suggest that RNA editing controls the function of one segment of the kaposin transcript, such that it has transforming activity during latent replication and possibly another, as-yet-undetermined, function during lytic replication. PMID- 17913831 TI - Histological diagnosis of prostate cancer in Korean men aged 70-79 years. AB - BACKGROUND: The objective of this study was to evaluate the value of the prostate specific antigen (PSA) in the diagnosis of prostate cancer in elderly Korean men, aged 70-79 years. METHODS: Patients with an abnormal digital rectal examination (DRE) and/or a serum PSA level greater than 2.0 ng/ml underwent a biopsy. A total of 344 men (median age 73 years) constituted the study cohort. RESULTS: Of 344 men, 163 (47.4%) were diagnosed with prostate cancer upon initial biopsy. The positive predictive value (PPV) for cancer was 48.4% for a PSA cutoff of 4 ng/ml, 65.3% for a cutoff of 10 ng/ml, and 87.0% for a cutoff of 20 ng/ml. When combined with an abnormal DRE, the predictive values for these PSA cutoffs increased to 79.3, 87.3 and 100%, respectively. When 10 ng/ml was chosen as a PSA cutoff level, about 50% of patients were found to have a Gleason score of 7 or higher. When 4 ng/ml was chosen as a PSA cutoff level, more than 50% of patients with an abnormal DRE were found to have a Gleason score of 7 or higher. CONCLUSIONS: In elderly men, more than 50% of patients are found to have cancers with a Gleason score of 7 or higher when their PSA level is greater than 10 ng/ml. This threshold may be lowered to 4 ng/ml in the presence of an abnormal DRE. Our findings provide a rationale for recommending a prostate biopsy in elderly patients with an abnormal DRE and/or an elevated serum PSA level. However, at present, it is not clear whether elderly men have better outcomes when they undergo cancer screening. PMID- 17913830 TI - Inhibition of alpharetrovirus replication by a range of human APOBEC3 proteins. AB - The mammalian APOBEC3 family of cytidine deaminases includes members that can act as potent inhibitors of retroviral infectivity and retrotransposon mobility. Here, we have examined whether the alpharetrovirus Rous sarcoma virus (RSV) is susceptible to inhibition by a range of human APOBEC3 proteins. We report that RSV is highly susceptible to inhibition by human APOBEC3G, APOBEC3F, and APOBEC3B and moderately susceptible to inhibition by human APOBEC3C and APOBEC3A. For all five proteins, inhibition of RSV infectivity was associated with selective virion incorporation and with C-to-T editing of the proviral DNA minus strand. In the case of APOBEC3G, editing appeared to be critical for effective inhibition. These data represent the first report of inhibition of retroviral infectivity and induction of proviral DNA editing by human APOBEC3A and reveal that alpharetroviruses, which do not normally encounter APOBEC3 proteins in their avian hosts, are susceptible to inhibition by all human APOBEC3 proteins tested. These data further suggest that the resistance of mammalian retroviruses to inhibition by the APOBEC3 proteins expressed in their normal host species is likely to have evolved subsequent to the appearance of this family of mammalian antiretroviral proteins some 35 million years ago; i.e., the base state of a naive retrovirus is susceptibility to inhibition. PMID- 17913832 TI - Role of prostaglandins in collecting duct-derived endothelin-1 regulation of blood pressure and water excretion. AB - Collecting duct (CD)-derived endothelin-1 (ET-1) exerts natriuretic, diuretic, and hypotensive effects. In vitro studies have implicated cyclooxygenase (COX) metabolites, and particularly PGE(2), as important mediators of CD ET-1 effects. However, it is unknown whether PGE(2) mediates CD-derived ET-1 actions in vivo. To test this, CD ET-1 knockout (KO) and control mice were studied. During normal salt and water intake, urinary PGE(2) excretion was unexpectedly increased in CD ET-1 KO mice compared with controls. Salt loading markedly increased urinary PGE(2) excretion in both groups of mice; however, the levels remained relatively higher in KO animals. Acutely isolated inner medullary collecting duct (IMCD) from KO mice also had increased PGE(2) production. The increased IMCD PGE(2) was COX-2 dependent, since NS-398 blocked all PGE(2) production. However, increased CD ET-1 KO COX-2 protein or mRNA could not be detected in inner medulla or IMCD, respectively. Inner medullary COX-1 mRNA and protein levels and IMCD COX-1 mRNA levels were unaffected by Na intake or CD ET-1 KO. KO mice on a normal or high-Na diet had elevated blood pressure compared with controls; this difference was not altered by indomethacin or NS-398 treatment. However, indomethacin or NS-398 did increase urine osmolality and reduce urine volume in KO, but not control, animals. In summary, IMCD COX-2-dependent PGE(2) production is increased in CD ET 1 KO mice, indicating that CD-derived ET-1 is not a primary regulator of IMCD PGE(2). Furthermore, the increased PGE(2) in CD ET-1 KO mice partly compensates for loss of ET-1 with respect to maintaining urinary water excretion, but not in blood pressure control. PMID- 17913833 TI - Purinergic control of apical plasma membrane PI(4,5)P2 levels sets ENaC activity in principal cells. AB - Activity of the epithelial sodium channel (ENaC) is limiting for Na(+) reabsorption at the distal nephron. Phosphoinositides, such as phosphatidylinositol 4,5-biphosphate [PI(4,5)P(2)] modulate the activity of this channel. Activation of purinergic receptors triggers multiple events, including activation of PKC and PLC, with the latter depleting plasma membrane PI(4,5)P(2). Here, we investigate regulation of ENaC in renal principal cells by purinergic receptors via PLC and PI(4,5)P(2). Purinergic signaling rapidly decreases ENaC open probability and apical membrane PI(4,5)P(2) levels with similar time courses. Moreover, inhibiting purinergic signaling with suramin rescues ENaC activity. The PLC inhibitor U73122, but not U73343, its inactive analog, recapitulates the action of suramin. In contrast, modulating PKC signaling failed to affect purinergic regulation of ENaC. Unexpectedly, inhibiting either purinergic receptors or PLC in resting cells dramatically increased ENaC activity above basal levels, indicating tonic activation of purinergic signaling in these polarized renal epithelial cells. Increased ENaC activity was associated with elevation of apical membrane PI(4,5)P(2) levels. Subsequent treatment with ATP in the presence of inhibited purinergic signaling failed to decrease ENaC activity and apical membrane PI(4,5)P(2) levels. Dwell-time analysis reveals that depletion of PI(4,5)P(2) forces ENaC toward a closed state. In contrast, increasing PI(4,5)P(2) levels above basal values locks the channel in an open state interrupted by brief closings. Thus our results suggest that purinergic control of apical membrane PI(4,5)P(2) levels is a major regulator of ENaC activity in renal epithelial cells. PMID- 17913834 TI - Association of VASP with TRPC4 in PKG-mediated inhibition of the store-operated calcium response in mesangial cells. AB - We tested the hypotheses that the NO-cGMP-PKG pathway mediates inhibition of the store-operated cation channel (SOC) in human glomerular mesangial cells (HMC) and that TRPC4, a molecular component of SOC in HMC, is associated with PKG phosphorylated vasodilator-stimulated phosphoprotein (VASP). Using fura 2 ratiometry, we measured intracellular Ca(2+) concentration [Ca(2+)](i) to determine whether sodium nitroprusside (SNP), an NO donor, and 8-Br-cGMP affected SOC-TRPC4 via PKG. We found that the SOC response in HMC was attenuated in the presence of 100 microM SNP, an NO donor, or 100 microM 8-Br-cGMP. Addition of DT 3 (2.5 microM), a specific PKG-1alpha inhibitor, reversed the effects of 8-Br cGMP on the SOC response. Application of 100 microM cAMP did not significantly inhibit the SOC response. RT-PCR and Western blotting revealed PKG-1alpha transcript and protein in HMC. Immunocytochemical analysis localized PKG-1alpha to the cytoplasm and plasma membrane of HMC. Previous studies have shown that PKG mediated phosphorylation of VASP attenuates cellular Ca(2+) entry, resulting in altered growth and proliferation. Therefore, we used Western blotting and immunocytochemistry to determine whether PKG-phosphorylated VASP associates with TRPC4. Western blot analysis revealed that 8-Br-cGMP enhanced the phosphorylation of VASP at serine 239 (Ser239), a known PKG phosphorylation site, in HMC within 5 min. Coimmunoprecipitation and coimmunostaining showed that P-Ser239-VASP associated with TRPC4. However, VASP that was unphosphorylated at Ser239 was not associated with TRPC4. These results indicate that VASP has a role in the NO/PKG 1alpha-mediated inhibition of the TRPC4-SOC response in HMC. PMID- 17913835 TI - Functional effects of nonsynonymous polymorphisms in the human TRPV1 gene. AB - The prototypical member of the vanilloid-responsive-like subfamily of transient receptor potential (TRP) channels is TRPV1. TRPV1 mediates aspects of nociception and neurogenic inflammation; however, new roles are emerging in sensation of both luminal stretch and systemic tonicity. Although at least six nonsynonymous polymorphisms in the human TRPV1 gene have been identified, there has been no systematic investigation into their functional consequences. When heterologously expressed in HEK293 cells, all variants exhibited equivalent EC(50) for the classic agonist capsaicin. This agonist elicited a greater maximal response in TRPV1(I315M) and TRPV1(P91S) variants (relative to TRPV1(WT)), as did a second agonist, anandamide. Expression of these two variants in whole-cell lysates and at the cell surface was markedly greater than that of wild-type TRPV1, whereas expression at the mRNA level was either unchanged (TRPV1(P91S)) or only very modestly increased (TRPV1(I315M)). Incorporation of multiple nonsynonymous SNPs, informed by the population-specific haplotype block structure of the TRPV1 gene, did not lead to variant channels with unique features vis-a-vis capsaicin responsiveness. Recently, polymorphisms/mutations were identified in two highly conserved TRPV1 residues in the nonobese diabetic (NOD) murine model. Incorporation of these changes into human TRPV1 gave rise to a channel with a normal EC(50) for capsaicin, but with a markedly elevated Hill slope such that the variant channel was hyporesponsive to capsaicin at low doses (<10 nM) and hyperresponsive at high doses (>10 nM). In aggregate, these data underscore expression-level and functional differences among naturally occurring TRPV1 variants; the implications with respect to human physiology are considered. PMID- 17913836 TI - Role of endogenously secreted angiotensin II in the CO2-induced stimulation of HCO3 reabsorption by renal proximal tubules. AB - Previous studies demonstrated that the proximal tubule (PT) responds to isolated increases in basolateral ([CO(2)](BL)) or "bath" CO(2) concentration by increasing the HCO(3)(-) reabsorption rate (J(HCO(3))). Blockade of the rabbit apical AT(1) receptor or knockout of the mouse AT(1A) receptor eliminates these effects, demonstrating a requirement for luminal ANG II that the PT itself synthesizes. In the present study, we examined the effects of the ACE inhibitor lisinopril on J(HCO(3)) in isolated perfused rabbit PTs (S2 segment), using out of-equilibrium solutions to make isolated changes in [CO(2)](BL) at a fixed baseline HCO(3)(-) concentration of 22 mM and fixed baseline pH of 7.4. Adding 60 or 240 nM lisinopril (in vitro K(i): 0.5 or 1.2 nM) to the lumen had no effect. These results are not consistent with the hypothesis that the PT secretes either angiotensinogen or ANG I. However, adding 60 nM basolateral lisinopril significantly decreased J(HCO(3)) at a [CO(2)](BL) of 20%. Moreover, 240 nM basolateral lisinopril decreased baseline (i.e., at 5% CO(2)) J(HCO(3)) by one half and completely eliminated the response to altering [CO(2)](BL) from 0 to 20%, but left intact the stimulatory effect of 10(-11) M basolateral ANG II. At extremely high concentrations (i.e., 100 microM), luminal lisinopril replicated the effects of 240 nM basolateral lisinopril. Our data are consistent with the hypothesis that lisinopril readily crosses the basolateral (but not apical) membrane to block ACE in a vesicular compartment. We conclude that the isolated PT predominantly secretes preformed ANG II, rather than angiotensinogen or ANG I. PMID- 17913837 TI - Vasopressin receptor subtype 2 activation increases cell proliferation in the renal medulla of AQP1 null mice. AB - Aquaporin (AQP) 1 null mice have a defect in the renal concentrating gradient because of their inability to generate a hyperosmotic medullary interstitium. To determine the effect of vasopressin on renal medullary gene expression, in the absence of high local osmolarity, we infused 1-deamino-8-d-arginine vasopressin (dDAVP), a V(2) receptor (V(2)R)-specific agonist, in AQP1 null mice for 7 days. cDNA microarray analysis was performed on the renal medullary tissue, and 5,140 genes of the possible 12,000 genes on the array were included in the analysis. In the renal medulla of AQP1 null mice, 245 transcripts were identified as increased by dDAVP infusion and 200 transcripts as decreased (1.5-fold or more). Quantitative real-time PCR measurements confirmed the increases seen for cyclin D1, early growth response gene 1, and activating transcription factor 3, genes associated with changes in cell cycle/growth. Changes in mRNA expression were correlated with changes in protein expression by semiquantitative immunoblotting; cyclin D1 and ATF3 were increased significantly in abundance following dDAVP infusion in the renal medulla of AQP1 null mice (161 and 461%, respectively). A significant increase in proliferation of medullary collecting ducts cells, following V(2)R activation, was identified by proliferating cell nuclear antigen immunohistochemistry; colocalization studies with AQP2 indicated that the increase in proliferation was primarily observed in principal cells of the inner medullary collecting duct (IMCD). V(2)R activation, via dDAVP, increased AQP2 and AQP3 protein abundance in the cortical collecting ducts of AQP1 null mice. However, V(2)R activation did not increase AQP2 protein abundance in the IMCD of AQP1 null mice. PMID- 17913838 TI - Stable RNA interference of synaptotagmin I in PC12 cells results in differential regulation of transmitter release. AB - In sympathetic neurons, it is well-established that the neurotransmitters, norepinephrine (NE), neuropeptide Y (NPY), and ATP are differentially coreleased from the same neurons. In this study, we determined whether synaptotagmin (syt) I, the primary Ca(2+) sensor for regulated release, could function as the protein that differentially regulates release of these neurotransmitters. Plasmid-based RNA interference was used to specifically and stably silence expression of syt I in a model secretory cell line. Whereas stimulated release of NPY and purines was abolished, stimulated catecholamine (CA) release was only reduced by approximately 50%. Although expression levels of tyrosine hydroxylase, the rate limiting enzyme in the dopamine synthesis pathway, was unaffected, expression of the vesicular monoamine transporter 1 was reduced by 50%. To evaluate whether NPY and CAs are found within the same vesicles and whether syt I is found localized to each of these NPY- and CA-containing vesicles, we used immunocytochemistry to determine that syt I colocalized with large dense core vesicles, with NPY, and with CAs. Furthermore, both CAs and NPY colocalized with one another and with large dense core vesicles. Electron micrographs show that large dense core vesicles are synthesized and available for release in cells that lack syt I. These results are consistent with syt I regulating differential release of transmitters. PMID- 17913839 TI - Akt-interacting proteins: attractive opposites. focus on "Carboxy-terminal modulator protein induces Akt phosphorylation and activation, thereby enhancing antiapoptotic, glycogen synthetic, and glucose uptake pathways". PMID- 17913840 TI - Opposing roles of EGF in IFN-alpha-induced epithelial barrier destabilization and tissue repair. AB - Balance between damaging influences and repair mechanisms determines the degree of tissue deterioration by inflammatory and other injury processes. Destabilization of the proximal tubular barrier has been previously shown to be induced by IFN-alpha, a cytokine crucial for linking innate and adaptive immune responses. EGF was implicated in rescue mechanisms from renal injury. To study the interplay between the two processes, we determined if EGF can prevent IFN alpha-induced barrier permeabilization. EGF did not counteract but even exacerbated the IFN-alpha-induced decrease of transepithelial electrical resistance in LLC-PK1 monolayers. For this effect Erk1/2 activation was necessary, linking barrier regulation to EGF-induced cell cycle progression. In contrast to its damage-intensifying effect, EGF also facilitated the regeneration of epithelial barrier function after the termination of IFN-alpha treatment. This effect was not mediated by Erk1/2 activation or cell proliferation since U0126, an Erk1/2 inhibitor, did not prevent but ameliorated recovery. However, EGF accelerated the downregulation of caspase-3 in recovering cells. Similarly, a pan caspase inhibitor was able to block caspase activity and, concomitantly, promote restoration of barrier function. Thus, barrier repair might be linked to an EGF mediated antiapoptotic mechanism. EGF appears to sensitize epithelial cells to the detrimental effects of IFN-alpha but also helps to restore barrier function in the healing phase. The observed dual effect of EGF might be explained by the different impact of proproliferative and antiapoptotic signaling pathways during and after cytokine treatment. The timing of epithelial exposure to damaging agents and repair factors was identified as a crucial parameter determining tissue fate. PMID- 17913841 TI - Gene expression of Na+/H+ exchanger in zebrafish H+ -ATPase-rich cells during acclimation to low-Na+ and acidic environments. AB - In mammalian nephrons, most of the Na(+) and HCO(3)(-) is reabsorbed by proximal tubular cells in which the Na(+)/H(+) exchanger 3 (NHE3) is the major player. The roles of NHEs in Na(+) uptake/acid-base regulation in freshwater (FW) fish gills are still being debated. In the present study, functional genomic approaches were used to clone and sequence the full-length cDNAs of the nhe family from zebrafish (Danio rerio). A phylogenetic tree analysis of the deduced amino acid sequences showed that zNHE1-8 are homologous to their mammalian counterparts. By RT-PCR analysis and double/triple in situ hybridization/immunocytochemistry, only zebrafish NHE3b was expressed in zebrafish gills and was colocalized with V-H(+) ATPase but not with Na(+)-K(+)-ATPase, indicating that H(+)-ATPase-rich (HR) cells specifically express NHE3b. A subsequent quantitative RT-PCR analysis demonstrated that acclimation to low-Na(+) FW caused upregulation and downregulation of the expressions of znhe3b and zatp6v0c (H(+)-ATPase C-subunit), respectively, in gill HR cells, whereas acclimation to acidic FW showed reversed effects on the expressions of these two genes. In conclusion, both NHE3b and H(+) ATPase are probably involved in Na(+) uptake/acid-base regulation in zebrafish gills, like mammalian kidneys, but the partitioning of these two transporters may be differentially regulated depending on the environmental situation in which fish are acclimatized. PMID- 17913842 TI - Lin-7 targets the Kir 2.3 channel on the basolateral membrane via a L27 domain interaction with CASK. AB - Polarized expression of the Kir 2.3 channel in renal epithelial cells is influenced by the opposing activities of two different PDZ proteins. Mammalian Lin-7 (mLin-7) directly interacts with Kir 2.3 to coordinate basolateral membrane expression, whereas the tax interacting protein 1 (TIP-1), composed of a single PDZ domain, competes for interaction with mLin-7 and drives Kir 2.3 into the endocytic pathway. Here we show that the basolateral targeting function of mLin-7 depends on its L27 domain, which directs interaction with a cognate L27 domain in the basolateral membrane-anchoring protein, calcium/calmodulin-dependent serine protein kinase (CASK). In MDCK cells, the expression of an mLin-7 mutant that lacks the L27 domain displaced Kir 2.3 from the mLin-7/CASK complex and caused the channel to accumulate into large intracellular vesicles that partially colocalized with Rab-11. Conversely, transplantation of the mLin-7 L27 domain to TIP-1 conferred CASK interaction and basolateral targeting of Kir 2.3. Expression of the CASK L27 domain redistributed endogenous mLin-7 to an intracellular compartment and caused Kir 2.3 to accumulate in subapical endosomes. Taken together, these data support a model whereby mLin-7 acts as a PDZ-to-L27 adapter, mediating indirect association of Kir 2.3 with a basolateral membrane scaffold and thereby stabilizing Kir 2.3 at the basolateral membrane. PMID- 17913843 TI - A dibenzoylmethane derivative protects dopaminergic neurons against both oxidative stress and endoplasmic reticulum stress. AB - The enhancement of intracellular stresses such as oxidative stress and endoplasmic reticulum (ER) stress has been implicated in several neurodegenerative disorders including Parkinson's disease (PD). During a search for compounds that regulate ER stress, a dibenzoylmethane (DBM) derivative 14-26 (2,2'-dimethoxydibenzoylmethane) was identified as a novel neuroprotective agent. Analysis in SH-SY5Y cells and in PC12 cells revealed that the regulation of ER stress by 14-26 was associated with its anti-oxidative property. 14-26 prevented the production of reactive oxygen species (ROS) when the cells were exposed to oxidants such as hydrogen peroxide and 6-hydroxydopamine (6-OHDA) or an ER stressor brefeldin A (BFA). 14-26 also prevented ROS-induced damage in both the ER and the mitochondria, including the protein carbonylation in the microsome and the reduction of the mitochondrial membrane potential. Further examination disclosed the presence of the iron-chelating activity in 14-26. In vivo, 14-26 suppressed both oxidative stress and ER stress and prevented neuronal death in the substantia nigra pars compacta (SNpc) after injection of 6-OHDA in mice. These results suggest that 14-26 is an antioxidant that protects dopaminergic neurons against both oxidative stress and ER stress and could be a therapeutic candidate for the treatment of PD. PMID- 17913844 TI - Activation of ADP-ribosylation factor regulates biogenesis of the ATP7A containing trans-Golgi network compartment and its Cu-induced trafficking. AB - ATP7A (MNK) regulates copper homeostasis by translocating from a compartment localized within the trans-Golgi network to the plasma membrane (PM) in response to increased copper load. The mechanisms that regulate the biogenesis of the MNK compartment and the trafficking of MNK are unclear. Here we show that the architecture of the MNK compartment is linked to the structure of the Golgi ribbon. Depletion of p115 tethering factor, which causes fragmentation of the Golgi ribbon, also disrupts the MNK compartment. In p115-depleted cells, MNK localizes to punctate structures that pattern on Golgi ministacks dispersed throughout the cell. Despite altered localization MNK trafficking still occurs, and MNK relocates from and returns to the fragmented compartment in response to copper. We further show that the biogenesis of the MNK compartment requires activation of ADP-ribosylation factor (Arf)1 GTPase, shown previously to facilitate the biogenesis of the Golgi ribbon. Activation of cellular Arf1 is prevented by 1) expressing an inactive "empty" form of Arf (Arf1/N126I), 2) expressing an inactive form of GBF1 (GBF1/E794K), guanine nucleotide exchange factor for Arf1, or 3) treating cells with brefeldin A, an inhibitor of GBF1 that disrupts MNK into a diffuse pattern. Importantly, preventing Arf activation inhibits copper-responsive trafficking of MNK to the PM. Our findings support a model in which active Arf is essential for the generation of the MNK compartment and for copper-responsive trafficking of MNK from there to the PM. Our findings provide an exciting foundation for identifying Arf1 effectors that facilitate the biogenesis of the MNK compartment and MNK traffic. PMID- 17913845 TI - Proliferation capacity of the renal proximal tubule involves the bulk of differentiated epithelial cells. AB - We investigated the proliferative capacity of renal proximal tubular cells in healthy rats. Previously, we observed that tubular cells originate from differentiated cells. We now found 1) by application of bromo-deoxyuridine (BrdU) for 14 days and costaining for BrdU, and the G(1)-phase marker cyclin D1 that the bulk of cells in the S3 segment of juvenile rats were involved in proliferation; 2) that although the proliferation rate was about 10-fold higher in juvenile rats compared with adult rats, roughly 40% of S3 cells were in G(1) in both groups; 3) that after a strong mitotic stimulus (lead acetate), proliferation was similar in juveniles and adults; 4) that there was a high incidence of cyclin D1-positive cells also in the healthy human kidney; and 5) by labeling dividing cells with BrdU for 2 days before the application of lead acetate and subsequent costaining for BrdU and cell cycle markers, that, although a strong mitotic stimulus does not abolish the period of quiescence following division, it shortens it markedly. Thus the capacity of the proximal tubule to rapidly recruit cells into division relies on a large reserve pool of cells in G(1) and on the shortening of the obligatory period of quiescence that follows division. PMID- 17913846 TI - Dura mater-derived FGF-2 mediates mitogenic signaling in calvarial osteoblasts. AB - Although dura mater tissue is believed to have an important role in calvarial reossification in many in vivo studies, few studies have shown the direct effect of dura mater cells on osteoblasts. In addition, no reports have yet identified the potential factor(s) responsible for various biological activities exerted by dura mater on calvarial reossification (e.g., cell proliferation). In this study, we tested the effect of dura mater on calvarial-derived osteoblasts by performing both heterotypic coculture and by culturing osteoblast cells with conditioned media harvested from dura mater cells of juvenile (3-day-old) and adult (30-day old) mice. The results presented here demonstrate that cellular proliferation of juvenile osteoblast cells was significantly increased by juvenile dura mater either in the coculture system or when dura mater cell-conditioned medium was applied to the osteoblast cells. Moreover, high levels of FGF-2 protein were detected in juvenile dura mater cells and their conditioned medium. In contrast, low levels of FGF-2 protein were detected in adult dura mater cells, whereas FGF 2 protein was not detectable in their conditioned medium. Abrogation of the mitogenic effect induced by juvenile dura mater cell-conditioned medium was achieved by introducing a neutralizing anti-FGF-2 antibody, thus indicating that FGF-2 may be responsible for the mitogenic effect of the juvenile dura mater. Moreover, data obtained by exploring the three major FGF-2 signaling pathways further reinforced the idea that FGF-2 might be an important paracrine signaling factor in vivo supplied by the underlying dura mater to the overlying calvarial osteoblasts. PMID- 17913847 TI - Phenotypic properties of adult mouse intrinsic cardiac neurons maintained in culture. AB - Intrinsic cardiac neurons are core elements of a complex neural network that serves as an important integrative center for regulation of cardiac function. Although mouse models are used frequently in cardiovascular research, very little is known about mouse intrinsic cardiac neurons. Accordingly, we have dissociated neurons from adult mouse heart, maintained these cells in culture, and defined their basic phenotypic properties. Neurons in culture were primarily unipolar, and 89% had prominent neurite outgrowth after 3 days (longest neurite length of 258 +/- 20 microm, n = 140). Many neurites formed close appositions with other neurons and nonneuronal cells. Neurite outgrowth was drastically reduced when neurons were kept in culture with a majority of nonneural cells eliminated. This finding suggests that nonneuronal cells release molecules that support neurite outgrowth. All neurons in coculture showed immunoreactivity for a full complement of cholinergic markers, but about 21% also stained for tyrosine hydroxylase, as observed previously in sections of intrinsic cardiac ganglia from mice and humans. Whole cell patch-clamp recordings demonstrated that these neurons have voltage-activated sodium current that is blocked by tetrodotoxin and that neurons exhibit phasic or accommodating patterns of action potential firing during a depolarizing current pulse. Several neurons exhibited a fast inward current mediated by nicotinic ACh receptors. Collectively, this work shows that neurons from adult mouse heart can be maintained in culture and exhibit appropriate phenotypic properties. Accordingly, these cultures provide a viable model for evaluating the physiology, pharmacology, and trophic factor sensitivity of adult mouse cardiac parasympathetic neurons. PMID- 17913848 TI - Atherosclerosis-prone hemodynamics differentially regulates endothelial and smooth muscle cell phenotypes and promotes pro-inflammatory priming. AB - Atherosclerosis is an inflammatory disease that preferentially forms at hemodynamically compromised regions of altered shear stress patterns. Endothelial cells (EC) and smooth muscle cells (SMC) undergo phenotypic modulation during atherosclerosis. An in vitro coculture model was developed to determine the role of hemodynamic regulation of EC and SMC phenotypes in coculture. Human ECs and SMCs were plated on a synthetic elastic lamina and human-derived atheroprone, and atheroprotective shear stresses were imposed on ECs. Atheroprone flow decreased genes associated with differentiated ECs (endothelial nitric oxide synthase, Tie2, and Kruppel-like factor 2) and SMCs (smooth muscle alpha-actin and myocardin) and induced a proinflammatory phenotype in ECs and SMCs (VCAM-1, IL-8, and monocyte chemoattractant protein-1). Atheroprone flow-induced changes in SMC differentiation markers were regulated at the chromatin level, as indicated by decreased serum response factor (SRF) binding to the smooth muscle alpha-actin CC(a/T)(6)GG (CArG) promoter region and decreased histone H(4) acetylation. Conversely, SRF and histone H(4) acetylation were enriched at the c-fos promoter in SMCs. In the presence of atheroprotective shear stresses, ECs aligned with the direction of flow and SMCs aligned more perpendicular to flow, similar to in vivo vessel organization. These results provide a novel mechanism whereby modulation of the EC phenotype by hemodynamic shear stresses, atheroprone or atheroprotective, play a critical role in mechanical-transcriptional coupling and regulation of the SMC phenotype. PMID- 17913849 TI - A web-based tool for in silico biomarker discovery based on tissue-specific protein profiles in normal and cancer tissues. AB - Here we report the development of a publicly available Web-based analysis tool for exploring proteins expressed in a tissue- or cancer-specific manner. The search queries are based on the human tissue profiles in normal and cancer cells in the Human Protein Atlas portal and rely on the individual annotation performed by pathologists of images representing immunohistochemically stained tissue sections. Approximately 1.8 million images representing more than 3000 antibodies directed toward human proteins were used in the study. The search tool allows for the systematic exploration of the protein atlas to discover potential protein biomarkers. Such biomarkers include tissue-specific markers, cell type-specific markers, tumor type-specific markers, markers of malignancy, and prognostic or predictive markers of cancers. Here we show examples of database queries to generate sets of candidate biomarker proteins for several of these different categories. Expression profiles of candidate proteins can then subsequently be validated by examination of the underlying high resolution images. The present study shows examples of search strategies revealing several potential protein biomarkers, including proteins specifically expressed in normal cells and in cancer cells from specified tumor types. The lists of candidate proteins can be used as a starting point for further validation in larger patient cohorts using both immunological approaches and technologies utilizing more classical proteomics tools. PMID- 17913850 TI - What has proteomics accomplished? PMID- 17913851 TI - Breakpoint characterization: a new approach for segregation analysis of paracentric inversion in human sperm. AB - Paracentric inversions (PAI) are structural chromosomal rearrangements generally considered to be harmless. Nevertheless, cases of viable recombinants have been reported, indicating the interest of studying the meiotic behaviour of these chromosomal abnormalities. To date, the few studies reported have been performed using either the human-hamster fertilization system or fluorescence in situ hybridization with centromeric or telomeric DNA probes. In order to improve the assessment of meiotic segregation in PAI, we present a new strategy based on the use of bacterial artificial chromosome (BAC) probes which allow a precise localization of chromosome breakpoints and the identification of all meiotic products in human sperm. Sperm samples from carriers of an inv(5) and an inv(14) were used to test this new high-resolution procedure. PMID- 17913852 TI - Use of emergency transport by patients with cardiopathies: a focus group study. AB - BACKGROUND: The importance of emergency systems accessible by telephone for the early attention of patients with ischemic cardiopathy is well known. In Andalusia, this service is provided by calling 061. However, studies show an insufficient use of the existing emergency system in this type of patient. OBJECTIVE: To identify explanations related to the decision regarding the method of transport to health-care systems, by private means of transport or 061 services, for people with ischemic cardiopathy from the onset of symptoms until their arrival at the Emergency Department. METHODS: Eleven focal groups were held with subjects diagnosed with ischemic cardiopathy. The discussions were related to the method of transport (using 061 or their own means of transport to a major hospital or to a nearby health facility for onward transfer), depending on the existence of previous experience and distance to the hospital. RESULTS: The method of transport is related to the degree of ignorance about what is happening, perceptions regarding the fastest way to reach the hospital, people available around the patient when the event takes place, vehicle availability and possible stressful situations. CONCLUSIONS: This study provides information about the reasons for using or not using the emergency transport systems for these patients and understanding how decisions were made. The study's importance lies in the possibility of improving individuals' access to health care systems through education-based actions and a strategic information and training plan that targets patients, families and health professionals. PMID- 17913853 TI - Synergistic cytotoxicity between tamoxifen and the plant toxin persin in human breast cancer cells is dependent on Bim expression and mediated by modulation of ceramide metabolism. AB - Phytochemicals have provided an abundant source of novel therapeutics for the treatment of human cancers. We have previously described a novel plant toxin, persin, derived from avocado leaves, which has unique in vivo actions in the mammary epithelium and Bim-dependent, cytotoxic effects in human breast cancer cells in vitro. Compounds structurally similar to persin, such as the polyunsaturated fatty acid, conjugated linoleic acid, can attenuate steroid hormone receptor signaling and modulate the response of breast cancer cells to antiestrogens. Here, we provide evidence that persin may have similar effects by showing its potent proapoptotic synergy with the antiestrogen 4-hydroxytamoxifen. However, although persin transcriptionally down-regulates estrogen receptor (ER) expression, unlike conjugated linoleic acid, it also shows efficacy in ER negative breast cancer cells, both alone and in combination with 4 hydroxytamoxifen, whereas normal breast epithelial cells are unaffected, suggesting it may act via a distinct, ER-independent mechanism. These proapoptotic synergistic interactions are associated with increased de novo ceramide synthesis and are dependent on expression of the proapoptotic protein Bim. These data show that persin should be further investigated as a potential novel cancer therapeutic agent because it significantly enhances the sensitivity of breast cancer cells to the cytotoxic effects of tamoxifen, regardless of their ER status, while displaying apparent specificity for the malignant phenotype. PMID- 17913854 TI - Inactivation of NF-kappaB by 3,3'-diindolylmethane contributes to increased apoptosis induced by chemotherapeutic agent in breast cancer cells. AB - Constitutive activation of Akt or nuclear factor-kappaB (NF-kappaB) has been reported to play a role in de novo resistance of cancer cells to chemotherapeutic agents, which is a major cause of treatment failure in cancer chemotherapy. Previous studies have shown that 3,3'-diindolylmethane (DIM), a major in vivo acid-catalyzed condensation product of indole-3-carbinol, is a potent inducer of apoptosis, inhibitor of tumor angiogenesis, and inactivator of Akt/NF-kappaB signaling in breast cancer cells. However, little is known regarding the inactivation of Akt/NF-kappaB that leads to chemosensitization of breast cancer cells to chemotherapeutic agents, such as Taxotere. Therefore, we examined whether the inactivation Akt/NF-kappaB signaling caused by B-DIM could sensitize breast cancer cells to chemotherapeutic agents both in vitro and in vivo. MDA-MB 231 cells were simultaneously treated with 15 to 45 micromol/L B-DIM and 0.5 to 1.0 nmol/L Taxotere for 24 to 72 h. Cell growth inhibition assay, apoptosis assay, electrophoretic mobility shift assay, and Western blotting were done. The combination treatment of 30 micromol/L B-DIM with 1.0 nmol/L Taxotere elicited significantly greater inhibition of cell growth compared with either agent alone. The combination treatment induced greater apoptosis in MDA-MB-231 cells compared with single agents. Moreover, we found that NF-kappaB activity was significantly decreased in cells treated with B-DIM and Taxotere. We also have tested our hypothesis using transfection studies, followed by combination treatment with B DIM/Taxotere, and found that combination treatment significantly inhibited cell growth and induced apoptosis in MDA-MB-231 breast cancer cells mediated by the inactivation of NF-kappaB, a specific target in vitro and in vivo. These results were also supported by animal experiments, which clearly showed that B-DIM sensitized the breast tumors to Taxotere, which resulted in greater antitumor activity mediated by the inhibition of Akt and NF-kappaB. Collectively, our results clearly suggest that inhibition of Akt/NF-kappaB signaling by B-DIM leads to chemosensitization of breast cancer cells to Taxotere, which may contribute to increased growth inhibition and apoptosis in breast cancer cells. The data obtained from our studies could be a novel breakthrough in cancer therapeutics by using nontoxic agents, such as B-DIM, in combination with other conventional therapeutic agents, such as Taxotere. PMID- 17913855 TI - The relevance of estrogen receptor-beta expression to the antiproliferative effects observed with histone deacetylase inhibitors and phytoestrogens in prostate cancer treatment. AB - In the prostate, estrogen receptor beta (ERbeta), the preferred receptor for phytoestrogens, has features of a tumor suppressor. To investigate the mechanisms underlying the beneficial effects on prostate cancer of histone deacetylase inhibitor valproic acid (VPA) and phytoestrogen tectorigenin, we analyzed the expression of ERbeta after tectorigenin or VPA treatment. For further functional analysis, we knocked down ERbeta expression by RNA interference. LNCaP prostate cancer cells were treated with 5 mmol/L VPA or 100 micromol/L tectorigenin and transfected with small interfering RNA (siRNA) against ERbeta. Control transfections were done with luciferase (LUC) siRNA. Expression of ERbeta was assessed by Western blot. mRNA expression was quantitated by real-time reverse transcription-PCR. Expression of ERbeta mRNA and protein markedly increased after VPA or tectorigenin treatment. When ERbeta was knocked down by siRNA, the expression of prostate-derived Ets factor, prostate-specific antigen, prostate cancer-specific indicator gene DD3(PCA3), insulin-like growth factor-1 receptor, the catalytic subunit of the telomerase, and ERalpha was up-regulated and the tectorigenin effects were abrogated. ERbeta levels were diminished in prostate cancer and loss of ERbeta was associated with proliferation. Here, we show that siRNA-mediated knockdown of ERbeta increases the expression of genes highly relevant to tumor cell proliferation. In addition, we show that one prominent result of treatment with VPA or tectorigenin is the up-regulation of ERbeta resulting in antiproliferative effects. Thus, these drugs, by restoring the regulatory function of ERbeta in tumor cells, could become useful in the intervention of prostate cancer. PMID- 17913856 TI - Expression of epidermal growth factor (EGF)/transforming growth factor-alpha by human lung cancer cells determines their response to EGF receptor tyrosine kinase inhibition in the lungs of mice. AB - Epidermal growth factor receptor (EGFR) has been extensively targeted in the treatment of non-small cell lung cancer, producing responses in a small number of patients. To study the role of ligand expression in mediating response to EGFR antagonism, we injected NCI-H441 [EGFR and EGF/transforming growth factor-alpha (TGF-alpha) positive] or PC14-PE6 (EGFR positive and EGF/TGF-alpha negative) human lung adenocarcinoma cells into the lungs of nude mice. We randomized the mice to receive treatment with the EGFR tyrosine kinase inhibitors gefitinib or AEE788 or vehicle. Treatment of mice bearing NCI-H441 but not PC14-PE6 lung tumors resulted in a significant reduction in primary tumor growth, pleural effusion, and lymph node metastasis. Immunohistochemical analyses revealed that NCI-H441 and PC14-PE6 cells expressed EGFR but that the expression of EGF/TGF alpha was high in NCI-H441 cells and very low in PC14-PE6 cells. Consequently, EGFR was activated in both tumor and tumor-associated endothelial cells in the NCI-H441 tumors but not in the PC14-PE6 tumors. Antagonism of EGFR signaling by treatment of mice with AEE788 decreased proliferation and increased apoptosis of both tumor cells and tumor-associated endothelial cells in NCI-H441 tumors but not in PC14-PE6 tumors. However, after transfection of PC14-PE6 cells with TGF alpha, lung tumors derived from the transfected cells expressed and activated EGFR in both tumor and tumor-associated endothelial cells and tumors responded to treatment with AEE788. Collectively, these results strongly suggest that the response of human lung cancers growing orthotopically in mice to the inhibition of EGFR signaling is determined by ligand (EGF/TGF-alpha) expression by tumor cells. Our findings provide an additional explanation for the susceptibility of lung cancers to treatment with EGFR tyrosine kinase inhibitors. PMID- 17913858 TI - High prevalence of the CD14-159CC genotype in patients infected with severe acute respiratory syndrome-associated coronavirus. AB - To investigate whether genetic factors of innate immunity might influence susceptibility and/or progression in individuals infected with SARS, we evaluated the CD14 gene polymorphism in 198 Hong Kong blood donors and 152 Hong Kong severe acute respiratory syndrome (SARS) patients who were previously genotyped for FcgammaRIIA polymorphisms. The prevalence of the CD14-159CC polymorphism was significantly higher in the patients with severe SARS than in the those with mild SARS or controls (31% versus 15% [mild SARS] or 20% [controls]; mild SARS: P = 0.029; odds ratio, 2.74; 95% confidence interval, 1.15 to 6.57; controls, P = 0.04; odds ratio, 2.41; 95% confidence interval, 1.05 to 5.54), and both CD14 159CC and FcgammaRIIA-RR131 are risk genotypes for severe SARS-CoV infection. PMID- 17913857 TI - Inhibitory activity of cetuximab on epidermal growth factor receptor mutations in non small cell lung cancers. AB - Mutations in the kinase domain of the epidermal growth factor receptor (EGFR) were identified in approximately 15% of all patients with non-small cell lung cancer (NSCLC). These mutations have been established as an indicator of superior response to gefitinib and erlotinib, small molecule inhibitors of the EGFR kinase domain. Whether these mutations would also render patients more susceptible to treatment with cetuximab (Erbitux), an EGFR-neutralizing antibody, is yet to be determined. In this study, we attempted to evaluate the effect of cetuximab on several NSCLC lines harboring some of the more common EGFR mutations (L858R and delL747-T753insS), as well as the recently identified kinase inhibitor-resistant mutation, T790M. We could show that the kinase activity of the abovementioned EGFR mutants was hindered by cetuximab, as detected by both cell-based phosphorylation and proliferation assays. Interestingly, cetuximab also induced enhanced degradation of the EGFR mutants as compared with the wild-type receptor. Most importantly, cetuximab successfully inhibited the growth of NSCLC lines in xenograft models. These results indicate the promising potential of cetuximab as a regimen for patients with NSCLC bearing these mutations. PMID- 17913859 TI - Serological survey of Rickettsia japonica infection in dogs and cats in Japan. AB - Antibodies against Rickettsia japonica in 20 of 1,207 dogs and 5 of 584 cats in Japan were detected using immunofluorescence. Some antibody-positive animals were detected in Niigata and Kagawa Prefectures, areas in which Japanese spotted fever in human patients has never been identified. Some animals were positive for antibodies against other new Rickettsia species. PMID- 17913860 TI - Development and evaluation of an Influenza virus subtype H7N2 vaccine candidate for pandemic preparedness. AB - Influenza virus of the H7N2 subtype has been introduced into noncommercial poultry in the United States, and this probably resulted in incidents of transmission of H7N2 virus to humans, documented in 2002 and 2003. This virus could be considered a potential threat to public health if it acquired person-to person transmissibility. A favored approach for global pandemic preparedness includes development of prepandemic vaccines for any potential pandemic virus. To this end, we created a high-growth reassortant virus (H7N2-PR8) containing the genes for the hemagglutinin and the neuraminidase from a low-pathogenicity (H7N2) virus strain and the remaining six genes from a human vaccine strain (H1N1). The reassortant strain was evaluated to assess its antigenicity, safety, and protective efficacy using a mouse model. Antigenicity studies using ferret antibodies raised against H7N2-PR8 indicated that this virus confers broad cross reactivity with divergent H7 viruses of different years and lineages. Mice and chickens inoculated with high doses of H7N2-PR8 supported virus replication but survived, indicating that this virus is comparable to other avian viruses of low pathogenicity. To assess the protective efficacy of H7N2-PR8, mice were immunized with two doses of formalin-inactivated H7N2-PR8, alone or with alum. Vaccinated mice subsequently challenged with highly pathogenic viruses from homologous and heterologous lineages A/Canada/444/04 (H7N3) and A/Netherlands/219/03 (H7N7) showed pronounced reduction of wild-type virus replication. These studies indicate that H7N2-PR8 is immunogenic, safe, and protective in animal models; these are the essential attributes to qualify for phase I human clinical trials as a prepandemic vaccine. PMID- 17913862 TI - Development of a rapid and convenient method for determination of rubella virus specific immunoglobulin G avidity. AB - We describe here a rapid and semiautomated method for the determination of rubella virus immunoglobulin G (IgG) avidity with the VIDAS instrument. A total of 153 serum samples from persons with naturally acquired rubella virus infections (n = 98), from vaccinated persons (n = 44), and from patients with autoantibodies (n = 11) were included in this study. The rubella virus-specific IgG avidity assay we developed for the VIDAS instrument was evaluated by comparison with an in-house method. Results obtained with the VIDAS instrument allow considering this method valuable to help confirm or exclude acute primary infection or recent vaccination. PMID- 17913861 TI - Use of a standardized bovine serum panel to evaluate a multiplexed nonstructural protein antibody assay for serological surveillance of foot-and-mouth disease. AB - Liquid array technology has previously been used to show proof of principle of a multiplexed nonstructural protein serological assay to differentiate foot-and mouth disease virus-infected and vaccinated animals. The current multiplexed assay consists of synthetically produced peptide signatures 3A, 3B, and 3D and the recombinant protein signature 3ABC in combination with four controls. To determine the diagnostic specificity of each signature in the multiplex, the assay was evaluated against a naive population (n = 104) and a vaccinated population (n = 94). Subsequently, the multiplexed assay was assessed by using a panel of bovine sera generated by the World Reference Laboratory for foot-and mouth disease in Pirbright, United Kingdom. This serum panel has been used to assess the performance of other singleplex enzyme-linked immunosorbent assay (ELISA)-based nonstructural protein antibody assays. The 3ABC signature in the multiplexed assay showed performance comparable to that of a commercially available nonstructural protein 3ABC ELISA (Cedi test), and additional information pertaining to the relative diagnostic sensitivity of each signature in the multiplex was acquired in one experiment. The encouraging results of the evaluation of the multiplexed assay against a panel of diagnostically relevant samples promote further assay development and optimization to generate an assay for routine use in foot-and-mouth disease serological surveillance. PMID- 17913863 TI - Detection of histoplasma antigen by a quantitative enzyme immunoassay. AB - The second-generation Histoplasma antigen immunoassay is semiquantitative, expressing results as a comparison to a negative control, which requires repeat testing of the prior specimen with the current specimen to accurately determine a change in antigen. Reporting results in this manner often is confusing to the ordering physician and laboratory. Development of a quantitative assay could improve accuracy, reduce interassay variability, and eliminate the need to test the prior sample with the current sample in the same assay. Calibrators with known concentrations of Histoplasma antigen were used to quantitate antigen in specimens from patients with histoplasmosis and from controls. Samples from cases of disseminated histoplasmosis or other mycoses and controls were tested to evaluate the performance characteristics of the quantitative assay. Paired specimens were evaluated to determine if quantitation eliminated the need to test the current and prior specimens in the same assay to assess a change in antigen. The sensitivity in samples from patients with AIDS and disseminated histoplasmosis was 100% in urine and 92.3% in serum. Cross-reactions occurred in 70% of other endemic mycoses, but not in aspergillosis. Specificity was 99% in controls with community-acquired pneumonia, medical conditions in which histoplasmosis was excluded, or healthy subjects. A change in antigen level categorized as an increase, no change, or decrease based on antigen units determined in the same assay agreed closely with the category of change in nanograms/milliliter determined from testing current and prior specimens in different assays. Sensitivity, specificity, and interassay precision are excellent in the new third-generation quantitative Histoplasma antigen immunoassay. PMID- 17913864 TI - Use of Saccharomyces cerevisiae-expressed recombinant nucleocapsid protein to detect Hantaan virus-specific immunoglobulin G (IgG) and IgM in oral fluid. AB - Hantaan virus is the causative agent of severe hemorrhagic fever with renal syndrome. Clinical surveillance for Hantaan virus infection is unreliable, and laboratory verification is essential. The detection of virus-specific immunoglobulin M (IgM) and IgG in serum is most commonly used for the diagnosis of hantavirus infection. Testing of oral fluid samples instead of serum offers many advantages for surveillance. However, commercial tests for hantavirus specific antibodies are unavailable. For the detection of Hantaan virus in the oral fluid of humans, we have developed a monoclonal antibody-based capture enzyme-linked immunosorbent IgM assay (IgM capture ELISA) and indirect enzyme linked immunosorbent IgG and IgM assays (indirect IgG and IgM ELISAs) for paired serum and oral fluid samples using the Saccharomyces cerevisiae yeast-expressed nucleocapsid protein of the Hantaan-Fojnica virus. The sensitivity and specificity of the oral fluid IgM capture ELISA in comparison with the results of the serum Hantaan virus IgM assay were 96.7% and of 94.9%, respectively. Thus, data on the overall performance of the oral fluid IgM capture ELISA are in close agreement with those of the serum IgM assay, and the method exhibits the potential to serve as an easily transferable tool for large-scale epidemiological studies. Data on the indirect IgM ELISA also showed close agreement with the serum IgM assay data; however, the indirect IgG ELISA displayed a lower sensitivity and a lower specificity. In conclusion, the IgM capture ELISA can be used with oral fluid instead of serum samples for the diagnosis of Hantaan virus infection. PMID- 17913865 TI - Immunity to Neisseria meningitidis group B in adults despite lack of serum bactericidal antibody. AB - Serum-complement-mediated bactericidal antibody (SBA) remains the serologic hallmark of protection against meningococcal disease, despite experimental and epidemiologic data that SBA may underestimate immunity. We measured bactericidal activity against three strains of Neisseria meningitidis group B in sera from 48 healthy adults and in whole blood from 15 subjects. Blood was anticoagulated with lepirudin, a specific thrombin inhibitor not known to activate complement. Depending on the test strain, protective SBA titers of >/=1:4 were present in only 8 to 15% of the subjects, whereas bactericidal activity was present in 40 to 87% of subjects according to the blood assay. Among SBA-negative subjects, blood from 23 to 42% gave a decrease of >/=2 log(10) CFU/ml after 1 h of incubation, and blood from 36 to 83% gave a decrease of >/=1 log(10) after 2 h. For most blood samples, bactericidal antibodies primarily were directed against noncapsular antigens, since activity was not inhibited by group B polysaccharide. For some SBA-negative subjects, white cells were not needed, since similar respective bactericidal activities were observed in blood and plasma. Bactericidal activity by whole blood of SBA-negative subjects can be rapid (<1 h) and effective (>/=2 log(10)) and, among all subjects, was four- to sixfold more prevalent than a positive SBA. Thus, while an SBA titer of >/=1:4 predicts protection against meningococcal disease, a titer of <1:4 is poorly predictive of susceptibility. More sensitive assays than SBA are needed to assess protective meningococcal immunity, or we risk underestimating the extent of immunity in the population and the effectiveness of new meningococcal vaccines. PMID- 17913866 TI - Effect of sexually transmitted disease (STD) coinfections on performance of three commercially available immunosorbent assays used for detection of herpes simplex virus type 2-specific antibody in men attending Baltimore, Maryland, STD clinics. AB - Two hundred seventy-nine serum samples from men attending sexually transmitted disease (STD) clinics in Baltimore, Maryland, were tested for herpes simplex virus type 2 (HSV-2)-specific antibody by three immunosorbent glycoprotein G-2 based assays (the Kalon, Focus, and Biokit assays). The results for all samples with positive results were confirmed by Western blotting (91/279; 32.6% HSV-2 seroprevalence). All patients were also tested for Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Mycoplasma genitalium, human immunodeficiency virus type 1, and hepatitis C virus. The Kalon assay performed very well with samples from this population (90.8% sensitive, 99.4% specific), whereas the Focus assay had a sensitivity (82.6%) much lower than that shown previously. For 19.7% of the samples, the Biokit assay gave an indeterminate result. It was found that the odds of a sample having a Biokit assay indeterminate result compared to that of having a definitive positive or negative results were 3.88 times greater for subjects concurrently infected with N. gonorrhoeae, after the effects of other STDs were controlled for (P = 0.001; 95% confidence interval, 1.78, 8.45). Unfortunately, we were unable to control for HSV-1 infection status in the regression model, which, on the basis of chi(2) analysis, might also affect the clarity of the Biokit test. The recommended index cutoff value of 1.1 for the Focus and Kalon assays was found to be optimal for this population. PMID- 17913867 TI - IMCL area density, but not IMCL utilization, is higher in women during moderate intensity endurance exercise, compared with men. AB - Women use more fat during endurance exercise as evidenced by a lower respiratory exchange ratio (RER). The contribution of intramyocellular lipid (IMCL) to lipid oxidation during endurance exercise is controversial, and studies investigating sex differences in IMCL utilization have found conflicting results. We determined the effect of sex on net IMCL use during an endurance exercise bout using an ultrastructural evaluation. Men (n = 17) and women (n = 19) completed 90-min cycling at 63% Vo(2peak). Biopsies were taken before and after exercise and fixed for electron microscopy to determine IMCL size, # IMCL/area, IMCL area density, and the % IMCL touching mitochondria. Women had a lower RER and carbohydrate oxidation rate and a higher lipid oxidation rate during exercise (P < 0.05), compared with men. Women had a higher # IMCL/area and IMCL area density (P < 0.05), compared with men. Women, but not men, had a higher % IMCL touching mitochondria postexercise (P = 0.03). Exercise decreased IMCL area density (P = 0.01), due to a decrease in the # IMCL/area (P = 0.02). There was no sex difference in IMCL size or net use. In conclusion, women have higher IMCL area density compared with men, due to an increased # IMCL and not an increased IMCL size, as well as an increased % IMCL touching mitochondria postexercise. Endurance exercise resulted in a net decrease in IMCL density due to decreased number of IMCL, not decreased IMCL size, in both sexes. PMID- 17913868 TI - Temperature and state dependence of dynamic phrenic oscillations in the decerebrate juvenile rat. AB - The aim of the present study was to determine characteristics of fast oscillations in the juvenile rat phrenic nerve (Ph) and to establish their temperature and state dependence. Two different age-matched decerebrate, baro- and chemodenervated rat preparations, in vivo and in situ arterially perfused models, were used to examine three systemic properties: 1) generation and dynamics of fast oscillations in Ph activity (both preparations), 2) responses to anoxia (both preparations), and 3) the effects of temperature on fast oscillations (in situ only). Both juvenile preparations generated power and coherence in two major bands analogous to adult medium- and high-frequency oscillations (HFO) at frequencies that increased with temperature but were lower than in adults. At < 28 degrees C, however, Ph oscillations were confined primarily to one low-frequency band (20-45 Hz). During sustained anoxia, both preparations produced stereotypical state changes from eupnea to hyperpnea to transition bursting (a behavior present only in vivo during incomplete ischemia) to gasping. Thus the juvenile rat produces a sequential pattern of responses to anoxia that are intermediate forms between those produced by neonates and those produced by adults. Time-frequency analysis determined that fast oscillations demonstrated dynamics over the course of the inspiratory burst and a state dependence similar to that of adults in vivo in which hyperpnea (and transition) bursts are associated with increases in HFO, while gasping contains no HFO. Our results confirm that both the fast oscillations in Ph activity and the coherence between Ph pairs produced by the juvenile rat are profoundly state- and temperature-dependent. PMID- 17913869 TI - Lipolysis and glycerol gluconeogenesis in simultaneously fasting and lactating northern elephant seals. AB - Adult female elephant seals (Mirounga angustirostris) combine long-term fasting with lactation and molting. Glycerol gluconeogenesis has been hypothesized as potentially meeting all of the glucose requirements of the seals during these fasts. To test this hypothesis, a primed constant infusion of [2-(14)C]glycerol was administered to 10 ten adult female elephant seals at 5 and 21-22 days postpartum and to 10 additional adult females immediately after the molt. Glycerol kinetics, rates of lipolysis, and the contribution of glycerol to glucose production were determined for each period. Plasma metabolite levels as well as insulin, glucagon, and cortisol were also measured. Glycerol rate of appearance was not significantly correlated with mass (P = 0.14, r2 = 0.33) but was significantly related to the percentage of glucose derived from glycerol (P < 0.01, r2 = 0.81) during late lactation. The contribution of glycerol to glucose production was <3% during each fasting period, suggesting a lower contribution to gluconeogenesis than is observed in other long-term fasting mammals. Because of a high rate of endogenous glucose production in fasting elephant seals, it is likely that glycerol gluconeogenesis still makes a substantial contribution to the substrate needs of glucose-dependent tissues. The lack of a relationship between glucoregulatory hormones and glycerol kinetics, glycerol gluconeogenesis, and metabolites supports the proposition that fasting elephant seals do not conform to the traditional insulin-glucagon model of substrate metabolism. PMID- 17913870 TI - Epidermal growth factor and sphingosine-1-phosphate stimulate Na+/H+ exchanger activity in the human placental syncytiotrophoblast. AB - The Na+/H+ exchanger (NHE) has a key role in intracellular pH ([pH]i) regulation of the syncytiotrophoblast in the human placenta and may have a role in the life cycle of this cell. In other cells the NHE (actually a family of up to 9 isoforms) is regulated by a variety of factors, but its regulation in the syncytiotrophoblast has not been studied. Here, we tested the hypotheses that EGF and sphingosine-1-phosphate (S1P), both of which affect trophoblast apoptosis and, in other cell types, NHE activity, stimulate syncytiotrophoblast NHE activity. Villous fragments from term human placentas were loaded with the pH sensitive dye, BCECF. NHE activity was measured by following the recovery of syncytiotrophoblast [pH]i following an imposed acid load, in the presence and absence of EGF, S1P, and specific inhibitors of NHE activity. Both EGF and S1P caused a dose-dependent upregulation of NHE activity in the syncytiotrophoblast. These effects were blocked by amiloride 500 microM (a nonspecific NHE blocker) and HOE694 100 microM (NHE blocker with NHE1 and 2 isoform selectivity). Effects of EGF were also reduced by the NHE3 selective blocker S3226 (used at 1 microM). These data provide the first evidence that both EGF and S1P stimulate NHE activity in the syncytiotrophoblast; they appear to do so predominantly by activating the NHE1 isoform. PMID- 17913871 TI - Baroreflexes of the rat. V. Tetanus-induced potentiation of ADN A-fiber responses at the NTS. AB - In a long-term neuromuscular blocked (NMB) rat preparation, tetanic stimulation of the aortic depressor nerve (ADN) enhanced the A-fiber evoked responses (ERs) in the cardiovascular region, the nucleus of the solitary tract (dmNTS). The potentiation persisted for at least several hours and may be a mechanism for adaptive adjustment of the gain of the baroreflex, with functional implications for blood pressure regulation. Using a capacitance electrode, we selectively stimulated A-fibers and acquired a stable 10-h "A-fiber only" ER baseline at the dmNTS. Following baseline, an A+C-fiber activating tetanus was applied to the ADN. The tetanus consisted of 1,000 "high current" pulses (10 trains; 300 mus, 100 Hz, 1 s), with intertrain interval of 9 s. A 10-h A-fiber only posttetanic test phase repeated the stimulus pattern of the baseline. Fourteen tetanus experiments were done in 12 rats. Compared with the baseline before tetanus, the A-fiber ER magnitudes of posttetanus hours were larger [F(13, 247) = 3.407, P < .001]; additionally, the 10-h posttetanus magnitude slopes were more positive than during 10 h before tetanus (df = 13; t = -3.47; P < 0.005); thus, an ADN A+C fiber-activating tetanus produced increases in the magnitude of the A-fiber ERs in the dmNTS that persisted for several hours. In an additional rat, application of an NMDA receptor antagonist, prior to the tetanus, blocked the potentiation effect. The stimulus protocols, magnitude and duration of the effect, and pharmacology resemble associative long-term potentiation (LTP). PMID- 17913874 TI - Collective cell migration patterns: follow the leader. PMID- 17913873 TI - Microtubule-based nuclear movement occurs independently of centrosome positioning in migrating neurons. AB - During neuronal migration in the developing brain, it is thought that the centrosome precedes the nucleus and provides a cue for nuclear migration along the microtubules. In time-lapse imaging studies of radially migrating granule cells in mouse cerebellar slices, we observed that the movements of the nucleus and centrosome appeared to occur independently of each other. The nucleus often migrated ahead of the centrosome during its saltatory movement, negating the supposed role of the centrosome in pulling the nucleus. The nucleus was associated with dynamic microtubules enveloping the entire nucleus and stable microtubules extending from the leading process to the anterior part of the nucleus. Neither of these perinuclear microtubules converged at the centrosome. Disruption or excess formation of stable microtubules attenuated nuclear migration, indicating that the configuration of stable microtubules is crucial for nuclear migration. The inhibition of LIS1 function, a regulator of a microtubule motor dynein, specifically blocks nuclear migration without affecting the coupling of the centrosome and microtubules in the leading process, suggesting that movements of the nucleus and centrosome are differentially regulated by dynein motor function. Thus, the nucleus moves along the microtubules independently of the position of the centrosome in migrating neurons. PMID- 17913875 TI - Tracing the effects of the Little Ice Age in the tropical lowlands of eastern Mesoamerica. AB - The causes of late-Holocene centennial to millennial scale climatic variability and the impact that such variability had on tropical ecosystems are still poorly understood. Here, we present a high-resolution, multiproxy record from lowland eastern Mesoamerica, studied to reconstruct climate and vegetation history during the last 2,000 years, in particular to evaluate the response of tropical vegetation to the cooling event of the Little Ice Age (LIA). Our data provide evidence that the densest tropical forest cover and the deepest lake of the last two millennia were coeval with the LIA, with two deep lake phases that follow the Sporer and Maunder minima in solar activity. The high tropical pollen accumulation rates limit LIA's winter cooling to a maximum of 2 degrees C. Tropical vegetation expansion during the LIA is best explained by a reduction in the extent of the dry season as a consequence of increased meridional flow leading to higher winter precipitation. These results highlight the importance of seasonal responses to climatic variability, a factor that could be of relevance when evaluating the impact of recent climate change. PMID- 17913876 TI - Modeling a layer 4-to-layer 2/3 module of a single column in rat neocortex: interweaving in vitro and in vivo experimental observations. AB - We report a step in constructing an in silico model of a neocortical column, focusing on the synaptic connection between layer 4 (L4) spiny neurons and L2/3 pyramidal cells in rat barrel cortex. It is based first on a detailed morphological and functional characterization of synaptically connected pairs of L4-L2/3 neurons from in vitro recordings and second, on in vivo recordings of voltage responses of L2/3 pyramidal cells to current pulses and to whisker deflection. In vitro data and a detailed compartmental model of L2/3 pyramidal cells enabled us to extract their specific membrane resistivity ( approximately 16,000 ohms x cm(2)) and capacitance ( approximately 0.8 microF/cm(2)) and the spatial distribution of L4-L2/3 synaptic contacts. The average peak conductance per L4 synaptic contact is 0.26 nS for the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid and 0.2 nS for NMDA receptors. The in vivo voltage response for current steps was then used to calibrate the model for in vivo conditions in the Down state. Consequently, the effect of a single whisker deflection was modeled by converging, on average, 350 +/- 20 L4 axons onto the modeled L2/3 pyramidal cell. Based on values of synaptic conductance, the spatial distribution of L4 synapses on L2/3 dendrites, and the average in vivo spiking probability of L4 spiny neurons, the model predicts that the feed-forward L4-L2/3 connection on its own does not fire the L2/3 neuron. With a broader distribution in the number of L4 neurons or with slight synchrony among them, the L2/3 model does spike with low probability. PMID- 17913877 TI - Sticky overhangs enhance siRNA-mediated gene silencing. AB - siRNA delivery to cells offers a convenient and powerful means of gene silencing that bypasses several barriers met by gene delivery. However, nonviral vectors, and especially polymers, form looser complexes with siRNA than with plasmid DNA. As a consequence, exchange of siRNA for larger polymeric anions such as proteoglycans found outside cells and at their surface may occur and lower delivery. We show here that making siRNAs "gene-like," via short complementary A(5-8)/T(5-8) 3' overhangs, increases complex stability, and hence RNase protection and gene silencing in vitro up to 10-fold. After decomplexation in the cytoplasm, sticky siRNA (ssiRNA) concatemers fall apart. ssiRNAs are therefore not inducing antiviral responses, as shown by the absence of IFN-beta production. Finally, transfection experiments in the mouse lung show that ssiRNA should be particularly suited to silencing with linear polyethylenimine in vivo. PMID- 17913878 TI - Transcription factor expression in lipopolysaccharide-activated peripheral-blood derived mononuclear cells. AB - Transcription factors play a key role in integrating and modulating biological information. In this study, we comprehensively measured the changing abundances of mRNAs over a time course of activation of human peripheral-blood-derived mononuclear cells ("macrophages") with lipopolysaccharide. Global and dynamic analysis of transcription factors in response to a physiological stimulus has yet to be achieved in a human system, and our efforts significantly advanced this goal. We used multiple global high-throughput technologies for measuring mRNA levels, including massively parallel signature sequencing and GeneChip microarrays. We identified 92 of 1,288 known human transcription factors as having significantly measurable changes during our 24-h time course. At least 42 of these changes were previously unidentified in this system. Our data demonstrate that some transcription factors operate in a functional range below 10 transcripts per cell, whereas others operate in a range three orders of magnitude greater. The highly reproducible response of many mRNAs indicates feedback control. A broad range of activation kinetics was observed; thus, combinatorial regulation by small subsets of transcription factors would permit almost any timing input to cis-regulatory elements controlling gene transcription. PMID- 17913879 TI - Genetic triple dissociation reveals multiple roles for dopamine in reinforcement learning. AB - What are the genetic and neural components that support adaptive learning from positive and negative outcomes? Here, we show with genetic analyses that three independent dopaminergic mechanisms contribute to reward and avoidance learning in humans. A polymorphism in the DARPP-32 gene, associated with striatal dopamine function, predicted relatively better probabilistic reward learning. Conversely, the C957T polymorphism of the DRD2 gene, associated with striatal D2 receptor function, predicted the degree to which participants learned to avoid choices that had been probabilistically associated with negative outcomes. The Val/Met polymorphism of the COMT gene, associated with prefrontal cortical dopamine function, predicted participants' ability to rapidly adapt behavior on a trial-to trial basis. These findings support a neurocomputational dissociation between striatal and prefrontal dopaminergic mechanisms in reinforcement learning. Computational maximum likelihood analyses reveal independent gene effects on three reinforcement learning parameters that can explain the observed dissociations. PMID- 17913880 TI - Beta-nicotinamide adenine dinucleotide is an inhibitory neurotransmitter in visceral smooth muscle. AB - Peripheral inhibitory nerves are physiological regulators of the contractile behavior of visceral smooth muscles. One of the transmitters responsible for inhibitory neurotransmission has been reputed to be a purine, possibly ATP. However, the exact identity of this substance has never been verified. Here we show that beta-nicotinamide adenine dinucleotide (beta-NAD), an inhibitory neurotransmitter candidate, is released by stimulation of enteric nerves in gastrointestinal muscles, and the pharmacological profile of beta-NAD mimics the endogenous neurotransmitter better than ATP. Levels of beta-NAD in superfusates of muscles after nerve stimulation exceed ATP by at least 30-fold; unlike ATP, the release of beta-NAD depends on the frequency of nerve stimulation. beta-NAD is released from enteric neurons, and release was blocked by tetrodotoxin or omega-conotoxin GVIA. beta-NAD is an agonist for P2Y1 receptors, as demonstrated by receptor-mediated responses in HEK293 cells expressing P2Y1 receptors. Exogenous beta-NAD mimics the effects of the enteric inhibitory neurotransmitter. Responses to beta-NAD and inhibitory junction potentials are blocked by the P2Y1 selective antagonist, MRS2179, and the nonselective P2 receptor antagonists, pyridoxal phosphate 6-azophenyl-2',4'-disulfonic acid and suramin. Responses to ATP are not blocked by these P2Y receptor inhibitors. The expression of CD38 in gastrointestinal muscles, and specifically in interstitial cells of Cajal, provides a means of transmitter disposal after stimulation. beta-NAD meets the traditional criteria for a neurotransmitter that contributes to enteric inhibitory regulation of visceral smooth muscles. PMID- 17913881 TI - Protein stability imposes limits on organism complexity and speed of molecular evolution. AB - Classical population genetics a priori assigns fitness to alleles without considering molecular or functional properties of proteins that these alleles encode. Here we study population dynamics in a model where fitness can be inferred from physical properties of proteins under a physiological assumption that loss of stability of any protein encoded by an essential gene confers a lethal phenotype. Accumulation of mutations in organisms containing Gamma genes can then be represented as diffusion within the Gamma-dimensional hypercube with adsorbing boundaries determined, in each dimension, by loss of a protein's stability and, at higher stability, by lack of protein sequences. Solving the diffusion equation whose parameters are derived from the data on point mutations in proteins, we determine a universal distribution of protein stabilities, in agreement with existing data. The theory provides a fundamental relation between mutation rate, maximal genome size, and thermodynamic response of proteins to point mutations. It establishes a universal speed limit on rate of molecular evolution by predicting that populations go extinct (via lethal mutagenesis) when mutation rate exceeds approximately six mutations per essential part of genome per replication for mesophilic organisms and one to two mutations per genome per replication for thermophilic ones. Several RNA viruses function close to the evolutionary speed limit, whereas error correction mechanisms used by DNA viruses and nonmutant strains of bacteria featuring various genome lengths and mutation rates have brought these organisms universally approximately 1,000-fold below the natural speed limit. PMID- 17913882 TI - Deep-water kelp refugia as potential hotspots of tropical marine diversity and productivity. AB - Classic marine ecological paradigms view kelp forests as inherently temperate boreal phenomena replaced by coral reefs in tropical waters. These paradigms hinge on the notion that tropical surface waters are too warm and nutrient depleted to support kelp productivity and survival. We present a synthetic oceanographic and ecophysiological model that accurately identifies all known kelp populations and, by using the same criteria, predicts the existence of >23,500 km(2) unexplored submerged (30- to 200-m depth) tropical kelp habitats. Predicted tropical kelp habitats were most probable in regions where bathymetry and upwelling resulted in mixed-layer shoaling above the depth of minimum annual irradiance dose for kelp survival. Using model predictions, we discovered extensive new deep-water Eisenia galapagensis populations in the Galapagos that increased in abundance with increasing depth to >60 m, complete with cold-water flora and fauna of temperate affinities. The predictability of deep-water kelp habitat and the discovery of expansive deep-water Galapagos kelp forests validate the extent of deep-water tropical kelp refugia, with potential implications for regional productivity and biodiversity, tropical food web ecology, and understanding of the resilience of tropical marine systems to climate change. PMID- 17913883 TI - Sustainable health: a new dimension of sustainability science. PMID- 17913884 TI - The transcription elongation factor TFIIS is a component of RNA polymerase II preinitiation complexes. AB - In this article, we provide direct evidence that the evolutionarily conserved transcription elongation factor TFIIS functions during preinitiation complex assembly. First, we identified TFIIS in a mass spectrometric screen of RNA polymerase II (Pol II) preinitiation complexes (PICs). Second, we show that the association of TFIIS with a promoter depends on functional PIC components including Mediator and the SAGA complex. Third, we demonstrate that TFIIS is required for efficient formation of active PICs. Using truncation mutants of TFIIS, we find that the Pol II-binding domain is the minimal domain necessary to stimulate PIC assembly. However, efficient formation of active PICs requires both the Pol II-binding domain and the poorly understood N-terminal domain. Importantly, Domain III, which is required for the elongation function of TFIIS, is dispensable during PIC assembly. The results demonstrate that TFIIS is a PIC component that is required for efficient formation and/or stability of the complex. PMID- 17913885 TI - Coevolution of languages and genes on the island of Sumba, eastern Indonesia. AB - Numerous studies indicate strong associations between languages and genes among human populations at the global scale, but all broader scale genetic and linguistic patterns must arise from processes originating at the community level. We examine linguistic and genetic variation in a contact zone on the eastern Indonesian island of Sumba, where Neolithic Austronesian farming communities settled and began interacting with aboriginal foraging societies approximately 3,500 years ago. Phylogenetic reconstruction based on a 200-word Swadesh list sampled from 29 localities supports the hypothesis that Sumbanese languages derive from a single ancestral Austronesian language. However, the proportion of cognates (words with a common origin) traceable to Proto-Austronesian (PAn) varies among language subgroups distributed across the island. Interestingly, a positive correlation was found between the percentage of Y chromosome lineages that derive from Austronesian (as opposed to aboriginal) ancestors and the retention of PAn cognates. We also find a striking correlation between the percentage of PAn cognates and geographic distance from the site where many Sumbanese believe their ancestors arrived on the island. These language-gene geography correlations, unprecedented at such a fine scale, imply that historical patterns of social interaction between expanding farmers and resident hunter gatherers largely explain community-level language evolution on Sumba. We propose a model to explain linguistic and demographic coevolution at fine spatial and temporal scales. PMID- 17913886 TI - Kinetic insulation as an effective mechanism for achieving pathway specificity in intracellular signaling networks. AB - Intracellular signaling pathways that share common components often elicit distinct physiological responses. In most cases, the biochemical mechanisms responsible for this signal specificity remain poorly understood. Protein scaffolds and cross-inhibition have been proposed as strategies to prevent unwanted cross-talk. Here, we report a mechanism for signal specificity termed "kinetic insulation." In this approach signals are selectively transmitted through the appropriate pathway based on their temporal profile. In particular, we demonstrate how pathway architectures downstream of a common component can be designed to efficiently separate transient signals from signals that increase slowly over time. Furthermore, we demonstrate that upstream signaling proteins can generate the appropriate input to the common pathway component regardless of the temporal profile of the external stimulus. Our results suggest that multilevel signaling cascades may have evolved to modulate the temporal profile of pathway activity so that stimulus information can be efficiently encoded and transmitted while ensuring signal specificity. PMID- 17913887 TI - Protein kinase C delta is essential for optimal macrophage-mediated phagosomal containment of Listeria monocytogenes. AB - Activation of macrophages and subsequent "killing" effector functions against infectious pathogens are essential for the establishment of protective immunity. NF-IL6 is a transcription factor downstream of IFN-gamma and TNF in the macrophage activation pathway required for bacterial killing. Comparison of microarray expression profiles of Listeria monocytogenes (LM)-infected macrophages from WT and NF-IL6-deficient mice enabled us to identify candidate genes downstream of NF-IL6 involved in the unknown pathways of LM killing independent of reactive oxygen intermediates and reactive nitrogen intermediates. One differentially expressed gene, PKCdelta, had higher mRNA levels in the LM infected NF-IL6-deficient macrophages as compared with WT. To define the role of PKCdelta during listeriosis, we infected PKCdelta-deficient mice with LM. PKCdelta-deficient mice were highly susceptible to LM infection with increased bacterial burden and enhanced histopathology despite enhanced NF-IL6 mRNA expression. Subsequent studies in PKCdelta-deficient macrophages demonstrated that, despite elevated levels of proinflammatory cytokines and NO production, increased escape of LM from the phagosome into the cytoplasm and uncontrolled bacterial growth occurred. Taken together these data identified PKCdelta as a critical factor for confinement of LM within macrophage phagosomes. PMID- 17913888 TI - Dentate gyrus-specific manipulation of beta-Ca2+/calmodulin-dependent kinase II disrupts memory consolidation. AB - Although the functions of alpha-Ca(2+)/calmodulin-dependent kinase II (CaMKII) have been studied extensively, the role of betaCaMKII, a coconstituent of the CaMKII holoenzyme in synaptic plasticity, learning, and memory has not been examined in vivo. Here we produce a transgenic mouse line in which the inducible and reversible manipulation of betaCaMKII activity is restricted to the hippocampal dentate gyrus, the region where long-term potentiation was originally discovered. We demonstrate that betaCaMKII activity in the dentate gyrus selectively impaired long-term potentiation in the dentate perforant path, but not in the CA1 Schaffer collateral pathway. Although the transgenic mice showed normal 1-day memories, they were severely impaired in 10-day contextual fear memory. Systematic manipulations of dentate betaCaMKII activity during various distinct memory stages further reveal the initial day within the postlearning consolidation period as a critical time window that is highly sensitive to changes in betaCaMKII activity. This study provides evidence not only for the functional role of betaCaMKII in the dentate gyrus plasticity and hippocampal memory, but also for the notion that the mismatch between the actual learning pattern and reactivation patterns in the dentate gyrus circuit can underlie long term memory consolidation. PMID- 17913889 TI - Z-ring force and cell shape during division in rod-like bacteria. AB - The life cycle of bacterial cells consists of repeated elongation, septum formation, and division. Before septum formation, a division ring called the Z ring, which is made of a filamentous tubulin analog, FtsZ, is seen at the mid cell. Together with several other proteins, FtsZ is essential for cell division. Visualization of strains with GFP-labeled FtsZ shows that the Z-ring contracts before septum formation and pinches the cell into two equal halves. Thus, the Z ring has been postulated to act as a force generator, although the magnitude of the contraction force is unknown. In this article, we develop a mathematical model to describe the process of growth and Z-ring contraction in rod-like bacteria. The elasticity and growth of the cell wall is incorporated in the model to predict the contraction speed, the cell shape, and the contraction force. With reasonable parameters, the model shows that a small force from the Z-ring (8 pN in Escherichia coli) is sufficient to accomplish division. PMID- 17913890 TI - Mortality traps and the dynamics of health transitions. AB - An examination of life expectancy in 1963 reveals twin peaks in the empirical distribution across countries: one group of countries clustered around a life expectancy of 40 years and a second group clustered around a life expectancy of 65 years. By 2003, the mode of each cluster had moved up by approximately 10 years. Although the two groups are similar in that within each of them, there is progress toward higher life expectancy, a number of countries appear to have made the jump from the high-mortality cluster to the low-mortality cluster. We reject the hypothesis that these changes reflect a simple convergence process. The data instead suggest continuous advances among many countries within clusters, with advances in life expectancy in some nations resulting in a jump from one cluster to the other. PMID- 17913891 TI - Chimeric constructs endow the human CFTR Cl- channel with the gating behavior of murine CFTR. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is a Cl(-) channel gated by ATP-driven nucleotide-binding domain (NBD) dimerization. Here we exploit species differences between human and murine CFTR to investigate CFTR channel gating. Using homologous recombination, we constructed human-murine CFTR (hmCFTR) chimeras with sequences from NBD1, NBD2, or the regulatory domain (RD) of human CFTR replaced by the equivalent regions of murine CFTR. The gating behavior of hmRD and human CFTR were indistinguishable, whereas hmNBD1 and hmNBD2 had subtle effects on channel gating, prolonging both burst duration and interburst interval. By contrast, hmNBD1+2, containing both NBDs of murine CFTR, reproduced the gating behavior of the subconductance state of murine CFTR, which has dramatically prolonged channel openings. The CFTR potentiator pyrophosphate (PP(i)) enhanced human, hmRD, and hmNBD1 CFTR Cl(-) currents, but not those of hmNBD2, hmNBD1+2, and murine CFTR. By analyzing the rate-equilibrium free-energy relationships of chimeric channels, we obtained snapshots of the conformation of the NBDs during ATP-driven dimerization. Our data demonstrate that the conformation of NBD1 changes before that of NBD2 during channel opening. This finding suggests that NBD dimerization does not proceed by a symmetric tweezer like motion, but instead in an asymmetric fashion led by NBD1. We conclude that the NBDs of murine CFTR determine the unique gating behavior of its subconductance state, whereas NBD2 controls channel potentiation by PP(i). PMID- 17913892 TI - Anorexia induced by activation of serotonin 5-HT4 receptors is mediated by increases in CART in the nucleus accumbens. AB - Anorexia nervosa is a growing concern in mental health, often inducing death. The potential neuronal deficits that may underlie abnormal inhibitions of food intake, however, remain largely unexplored. We hypothesized that anorexia may involve altered signaling events within the nucleus accumbens (NAc), a brain structure involved in reward. We show here that direct stimulation of serotonin (5-hydroxytryptamine, 5-HT) 4 receptors (5-HT(4)R) in the NAc reduces the physiological drive to eat and increases CART (cocaine- and amphetamine-regulated transcript) mRNA levels in fed and food-deprived mice. It further shows that injecting 5-HT(4)R antagonist or siRNA-mediated 5-HT(4)R knockdown into the NAc induced hyperphagia only in fed mice. This hyperphagia was not associated with changes in CART mRNA expression in the NAc in fed and food-deprived mice. Results include that 5-HT(4)R control CART mRNA expression into the NAc via a cAMP/PKA signaling pathway. Considering that CART may interfere with food- and drug related rewards, we tested whether the appetite suppressant properties of 3,4-N methylenedioxymethamphetamine (MDMA, ecstasy) involve the 5-HT(4)R. Using 5 HT(4)R knockout mice, we demonstrate that 5-HT(4)R are required for the anorectic effect of MDMA as well as for the MDMA-induced enhancement of CART mRNA expression in the NAc. Directly injecting CART peptide or CART siRNA into the NAc reduces or increases food consumption, respectively. Finally, stimulating 5 HT(4)R- and MDMA-induced anorexia were both reduced by injecting CART siRNA into the NAc. Collectively, these results demonstrate that 5-HT(4)R-mediated up regulation of CART in the NAc triggers the appetite-suppressant effects of ecstasy. PMID- 17913893 TI - TRPM7 channels in hippocampal neurons detect levels of extracellular divalent cations. AB - Exposure to low Ca(2+) and/or Mg(2+) is tolerated by cardiac myocytes, astrocytes, and neurons, but restoration to normal divalent cation levels paradoxically causes Ca(2+) overload and cell death. This phenomenon has been called the "Ca(2+) paradox" of ischemia-reperfusion. The mechanism by which a decrease in extracellular Ca(2+) and Mg(2+) is "detected" and triggers subsequent cell death is unknown. Transient periods of brain ischemia are characterized by substantial decreases in extracellular Ca(2+) and Mg(2+) that mimic the initial condition of the Ca(2+) paradox. In CA1 hippocampal neurons, lowering extracellular divalents stimulates a nonselective cation current. We show that this current resembles TRPM7 currents in several ways. Both (i) respond to transient decreases in extracellular divalents with inward currents and cell excitation, (ii) demonstrate outward rectification that depends on the presence of extracellular divalents, (iii) are inhibited by physiological concentrations of intracellular Mg(2+), (iv) are enhanced by intracellular phosphatidylinositol 4,5-bisphosphate (PIP(2)), and (v) can be inhibited by Galphaq-linked G protein coupled receptors linked to phospholipase C beta1-induced hydrolysis of PIP(2). Furthermore, suppression of TRPM7 expression in hippocampal neurons strongly depressed the inward currents evoked by lowering extracellular divalents. Finally, we show that activation of TRPM7 channels by lowering divalents significantly contributes to cell death. Together, the results demonstrate that TRPM7 contributes to the mechanism by which hippocampal neurons "detect" reductions in extracellular divalents and provide a means by which TRPM7 contributes to neuronal death during transient brain ischemia. PMID- 17913894 TI - Bridges to sustainable tropical health. AB - Ensuring sustainable health in the tropics will require bridge building between communities that currently have a limited track record of interaction. It will also require new organizational innovation if much of the negative health consequences of large-scale economic development projects are to be equitably mitigated, if not prevented. We focus attention on three specific contexts: (i) forging linkages between the engineering and health communities to implement clean water and sanitation on a broad scale to prevent reworming, after the current deworming-only programs, of people by diverse intestinal parasites; (ii) building integrated human and animal disease surveillance infrastructure and technical capacity in tropical countries on the reporting and scientific evidence requirements of the sanitary and phytosanitary agreement under the World Trade Organization; and (iii) developing an independent and equitable organizational structure for health impact assessments as well as monitoring and mitigation of health consequences of economic development projects. Effective global disease surveillance and timely early warning of new outbreaks will require a far closer integration of veterinary and human medicine than heretofore. Many of the necessary surveillance components exist within separate animal- and human oriented organizations. The challenge is to build the necessary bridges between them. PMID- 17913895 TI - Sustainable vector control and management of Chagas disease in the Gran Chaco, Argentina. AB - Chagas disease remains a serious obstacle to health and economic development in Latin America, especially for the rural poor. We report the long-term effects of interventions in rural villages in northern Argentina during 1984-2006. Two community-wide campaigns of residual insecticide spraying immediately and strongly reduced domestic infestation and infection with Trypanosoma cruzi in Triatoma infestans bugs and dogs and more gradually reduced the seroprevalence of children <15 years of age. Because no effective surveillance and control actions followed the first campaign in 1985, transmission resurged in 2-3 years. Renewed interventions in 1992 followed by sustained, supervised, community-based vector control largely suppressed the reestablishment of domestic bug colonies and finally led to the interruption of local human T. cruzi transmission. Human incidence of infection was nearly an order of magnitude higher in peripheral rural areas under pulsed, unsupervised, community-based interventions, where human transmission became apparent in 2000. The sustained, supervised, community based strategy nearly interrupted domestic transmission to dogs but did not eliminate T. infestans despite the absence of pyrethroid-insecticide resistance. T. infestans persisted in part because of the lack of major changes in housing construction and quality. Sustained community participation grew out of establishing a trusted relationship with the affected communities and the local schools. The process included health promotion and community mobilization, motivation, and supervision in close cooperation with locally nominated leaders. PMID- 17913897 TI - The neural substrate of disappointment revealed? PMID- 17913896 TI - Pharmacokinetic profile of temsirolimus with concomitant administration of cytochrome p450-inducing medications. AB - Temsirolimus is a novel inhibitor of the mammalian target of rapamycin, with antitumor activity in advanced tumors. Because temsirolimus and its metabolite, sirolimus, are cytochrome P450 (CYP) 3A4/5 substrates, the potential exists for interaction with drugs that induce CYP3A activity, including enzyme inducers and rifampin. Cancer patients received once-weekly intravenous (IV) 220 mg/m(2) temsirolimus with or without enzyme inducers. Coadministration with enzyme inducers decreased temsirolimus maximum plasma concentration (C(max)) by 36% and increased volume of distribution by 99%. Sirolimus C(max) and area under the concentration-time curve (AUC) were decreased by 67% and 43%, respectively. In healthy adult subjects, coadministration of 25-mg intravenous temsirolimus with rifampin had no significant effect on temsirolimus C(max) and AUC but decreased sirolimus C(max) and AUC by 65% and 56%, respectively. Rifampin decreased AUC(sum) by 41%. Temsirolimus was well tolerated in both studies. If concomitant agents with CYP3A induction potential are used, higher temsirolimus doses may be needed to achieve adequate tumor tissue drug levels. PMID- 17913898 TI - The influence of GABA on dendritic development in vivo. PMID- 17913899 TI - Modulation of the receptive fields of midbrain neurons elicited by thalamic electrical stimulation through corticofugal feedback. AB - The ascending and descending projections of the central auditory system form multiple tonotopic loops. This study specifically examines the tonotopic pathway from the auditory thalamus to the auditory cortex and then to the auditory midbrain in mice. We observed the changes of receptive fields in the central nucleus of the inferior colliculus of the midbrain evoked by focal electrical stimulation of the ventral division of the medial geniculate body of the thalamus. The receptive field of an auditory neuron was characterized by five parameters: the best frequency, minimum threshold, bandwidth, size of receptive field, and average spike number. We found that focal thalamic stimulation changed the parametric values characterizing the recorded collicular receptive fields toward those characterizing the stimulated thalamic receptive fields. Cortical inactivation with muscimol prevented the development of the collicular plasticity induced by focal thalamic stimulation. Our data suggest that the intact colliculo thalamo-cortico-collicular loops are important for the coordination of sound guided plasticity in the central auditory system. PMID- 17913900 TI - Supplementary motor area and presupplementary motor area: targets of basal ganglia and cerebellar output. AB - We used retrograde transneuronal transport of neurotropic viruses in Cebus monkeys to examine the organization of basal ganglia and cerebellar projections to two cortical areas on the medial wall of the hemisphere, the supplementary motor area (SMA) and the pre-SMA. We found that both of these cortical areas are the targets of disynaptic projections from the dentate nucleus of the cerebellum and from the internal segment of the globus pallidus (GPi). On average, the number of pallidal neurons that project to the SMA and pre-SMA is approximately three to four times greater than the number of dentate neurons that project to these cortical areas. GPi neurons that project to the pre-SMA are located in a rostral, "associative" territory of the nucleus, whereas GPi neurons that project to the SMA are located in a more caudal and ventral "sensorimotor" territory. Similarly, dentate neurons that project to the pre-SMA are located in a ventral, "nonmotor" domain of the nucleus, whereas dentate neurons that project to the SMA are located in a more dorsal, "motor" domain. The differential origin of subcortical projections to the SMA and pre-SMA suggests that these cortical areas are nodes in distinct neural systems. Although both systems are the target of outputs from the basal ganglia and the cerebellum, these two cortical areas seem to be dominated by basal ganglia input. PMID- 17913901 TI - Enhanced astrocytic Ca2+ signals contribute to neuronal excitotoxicity after status epilepticus. AB - Status epilepticus (SE), an unremitting seizure, is known to cause a variety of traumatic responses including delayed neuronal death and later cognitive decline. Although excitotoxicity has been implicated in this delayed process, the cellular mechanisms are unclear. Because our previous brain slice studies have shown that chemically induced epileptiform activity can lead to elevated astrocytic Ca2+ signaling and because these signals are able to induce the release of the excitotoxic transmitter glutamate from these glia, we asked whether astrocytes are activated during status epilepticus and whether they contribute to delayed neuronal death in vivo. Using two-photon microscopy in vivo, we show that status epilepticus enhances astrocytic Ca2+ signals for 3 d and that the period of elevated glial Ca2+ signaling is correlated with the period of delayed neuronal death. To ask whether astrocytes contribute to delayed neuronal death, we first administered antagonists which inhibit gliotransmission: MPEP [2-methyl-6 (phenylethynyl)pyridine], a metabotropic glutamate receptor 5 antagonist that blocks astrocytic Ca2+ signals in vivo, and ifenprodil, an NMDA receptor antagonist that reduces the actions of glial-derived glutamate. Administration of these antagonists after SE provided significant neuronal protection raising the potential for a glial contribution to neuronal death. To test this glial hypothesis directly, we loaded Ca2+ chelators selectively into astrocytes after status epilepticus. We demonstrate that the selective attenuation of glial Ca2+ signals leads to neuronal protection. These observations support neurotoxic roles for astrocytic gliotransmission in pathological conditions and identify this process as a novel therapeutic target. PMID- 17913902 TI - Brain-derived neurotrophic factor rescues synaptic plasticity in a mouse model of fragile X syndrome. AB - Mice lacking expression of the fragile X mental retardation 1 (Fmr1) gene have deficits in types of learning that are dependent on the hippocampus. Here, we report that long-term potentiation (LTP) elicited by threshold levels of theta burst afferent stimulation (TBS) is severely impaired in hippocampal field CA1 of young adult Fmr1 knock-out mice. The deficit was not associated with changes in postsynaptic responses to TBS, NMDA receptor activation, or levels of punctate glutamic acid decarboxylase-65/67 immunoreactivity. TBS-induced actin polymerization within dendritic spines was also normal. The LTP impairment was evident within 5 min of induction and, thus, may not be secondary to defects in activity-initiated protein synthesis. Protein levels for both brain-derived neurotrophic factor (BDNF), a neurotrophin that activates pathways involved in spine cytoskeletal reorganization, and its TrkB receptor were comparable between genotypes. BDNF infusion had no effect on baseline transmission or on postsynaptic responses to theta burst stimulation, but nonetheless fully restored LTP in slices from fragile X mice. These results indicate that the fragile X mutation produces a highly selective impairment to LTP, possibly at a step downstream of actin filament assembly, and suggest a means for overcoming this deficit. The possibility of a pharmacological therapy based on these results is discussed. PMID- 17913903 TI - Maternal immune activation alters fetal brain development through interleukin-6. AB - Schizophrenia and autism are thought to result from the interaction between a susceptibility genotype and environmental risk factors. The offspring of women who experience infection while pregnant have an increased risk for these disorders. Maternal immune activation (MIA) in pregnant rodents produces offspring with abnormalities in behavior, histology, and gene expression that are reminiscent of schizophrenia and autism, making MIA a useful model of the disorders. However, the mechanism by which MIA causes long-term behavioral deficits in the offspring is unknown. Here we show that the cytokine interleukin 6 (IL-6) is critical for mediating the behavioral and transcriptional changes in the offspring. A single maternal injection of IL-6 on day 12.5 of mouse pregnancy causes prepulse inhibition (PPI) and latent inhibition (LI) deficits in the adult offspring. Moreover, coadministration of an anti-IL-6 antibody in the poly(I:C) model of MIA prevents the PPI, LI, and exploratory and social deficits caused by poly(I:C) and normalizes the associated changes in gene expression in the brains of adult offspring. Finally, MIA in IL-6 knock-out mice does not result in several of the behavioral changes seen in the offspring of wild-type mice after MIA. The identification of IL-6 as a key intermediary should aid in the molecular dissection of the pathways whereby MIA alters fetal brain development, which can shed new light on the pathophysiological mechanisms that predispose to schizophrenia and autism. PMID- 17913906 TI - Interactions between circadian neurons control temperature synchronization of Drosophila behavior. AB - Most animals rely on circadian clocks to synchronize their physiology and behavior with the day/night cycle. Light and temperature are the major physical variables that can synchronize circadian rhythms. Although the effects of light on circadian behavior have been studied in detail in Drosophila, the neuronal mechanisms underlying temperature synchronization of circadian behavior have received less attention. Here, we show that temperature cycles synchronize and durably affect circadian behavior in Drosophila in the absence of light input. This synchronization depends on the well characterized and functionally coupled circadian neurons controlling the morning and evening activity under light/dark cycles: the M cells and E cells. However, circadian neurons distinct from the M and E cells are implicated in the control of rhythmic behavior specifically under temperature cycles. These additional neurons play a dual role: they promote evening activity and negatively regulate E cell function in the middle of the day. We also demonstrate that, although temperature synchronizes circadian behavior more slowly than light, this synchronization is considerably accelerated when the M cell oscillator is absent or genetically altered. Thus, whereas the E cells show great responsiveness to temperature input, the M cells and their robust self-sustained pacemaker act as a resistance to behavioral synchronization by temperature cycles. In conclusion, the behavioral responses to temperature input are determined by both the individual properties of specific groups of circadian neurons and their organization in a neural network. PMID- 17913904 TI - Inflammation activates the interferon signaling pathways in taste bud cells. AB - Patients with viral and bacterial infections or other inflammatory illnesses often experience taste dysfunctions. The agents responsible for these taste disorders are thought to be related to infection-induced inflammation, but the mechanisms are not known. As a first step in characterizing the possible role of inflammation in taste disorders, we report here evidence for the presence of interferon (IFN)-mediated signaling pathways in taste bud cells. IFN receptors, particularly the IFN-gamma receptor IFNGR1, are coexpressed with the taste cell type markers neuronal cell adhesion molecule and alpha-gustducin, suggesting that both the taste receptor cells and synapse-forming cells in the taste bud can be stimulated by IFN. Incubation of taste bud-containing lingual epithelia with recombinant IFN-alpha and IFN-gamma triggered the IFN-mediated signaling cascades, resulting in the phosphorylation of the downstream STAT1 (signal transducer and activator of transcription protein 1) transcription factor. Intraperitoneal injection of lipopolysaccharide or polyinosinic:polycytidylic acid into mice, mimicking bacterial and viral infections, respectively, altered gene expression patterns in taste bud cells. Furthermore, the systemic administration of either IFN-alpha or IFN-gamma significantly increased the number of taste bud cells undergoing programmed cell death. These findings suggest that bacterial and viral infection-induced IFNs can act directly on taste bud cells, affecting their cellular function in taste transduction, and that IFN induced apoptosis in taste buds may cause abnormal cell turnover and skew the representation of different taste bud cell types, leading to the development of taste disorders. To our knowledge, this is the first study providing direct evidence that inflammation can affect taste buds through cytokine signaling pathways. PMID- 17913905 TI - CNS-derived interleukin-4 is essential for the regulation of autoimmune inflammation and induces a state of alternative activation in microglial cells. AB - Regulation of inflammation in the CNS is essential to prevent irreversible cellular damage that can occur in neurodegenerative diseases such as multiple sclerosis (MS). We investigated the role of interleukin-4 (IL-4) in regulating CNS inflammation using the animal model of MS, experimental autoimmune encephalomyelitis (EAE). We found that CNS-derived IL-4 was a critical regulator because mice with a deficiency in IL-4 production in the CNS, but not the periphery, had exacerbated EAE associated with a significant increase in the absolute number of infiltrating inflammatory cells. We also found that CNS resident microglial cells in both the resting and activated state produced the protein Ym1, which is a marker of alternatively activated macrophages (aaMphis), in an IL-4-dependent manner. This aaMphi phenotype extended to the lack of nitric oxide (NO) production by activated microglial cells, which is a marker of classically activated macrophages. We also show that IL-4 induced the expression of Ym1 in peripheral infiltrating macrophages, which also produce NO. Thus, macrophages that migrate into the CNS exhibit a dual phenotype. These data indicate that IL-4 production in the CNS is essential for controlling autoimmune inflammation by inducing a microglial cell aaMphi phenotype. Macrophages that have undergone alternative activation have been shown to be important in tissue repair; thus, our results suggest a new role for microglial cells in the regulation of inflammation in the CNS. PMID- 17913907 TI - Preparatory activity in occipital cortex in early blind humans predicts auditory perceptual performance. AB - Early onset blindness leads to a dramatic alteration in the way the world is perceived, a change that is detectable in the organization of the brain. Several studies have confirmed that blindness leads to functional alterations in occipital cortices that normally serve visual functions. These reorganized brain regions respond to a variety of tasks and stimuli, but their specific functions are unclear. In sighted individuals, several studies have reported preparatory activity in retinotopic areas, which enhances perceptual sensitivity. "Baseline shifts," changes in activity associated with a cue predicting an upcoming event, provides a marker for attentional modulation. Here we demonstrate that, in early blind subjects, medial occipital areas produced significant blood oxygenation level-dependent (BOLD) responses to a cue signaling an auditory discrimination trial but not to a cue indicating a no-trial period. Furthermore, the amplitude of the BOLD response in the anterior calcarine sulcus of early blind subjects correlated with their discrimination performance on the auditory backward masking task. Preparatory BOLD responses also were present in auditory cortices, although they were more robust in blind than sighted control subjects. The pattern of response in visual areas is similar to preparatory effects observed during visual selective attention in sighted subjects and consistent with the hypothesis that the mechanisms implicated in visual attention continue to modulate occipital cortex in the early blind. A possible source of this top-down modulation may be the frontoparietal circuits that retain their connectivity with the reorganized occipital cortex and as a result influence processing of nonvisual stimuli in the blind. PMID- 17913908 TI - Single-neuron stability during repeated reaching in macaque premotor cortex. AB - Some movements that animals and humans make are highly stereotyped, repeated with little variation. The patterns of neural activity associated with repeats of a movement may be highly similar, or the same movement may arise from different patterns of neural activity, if the brain exploits redundancies in the neural projections to muscles. We examined the stability of the relationship between neural activity and behavior. We asked whether the variability in neural activity that we observed during repeated reaching was consistent with a noisy but stable relationship, or with a changing relationship, between neural activity and behavior. Monkeys performed highly similar reaches under tight behavioral control, while many neurons in the dorsal aspect of premotor cortex and the primary motor cortex were simultaneously monitored for several hours. Neural activity was predominantly stable over time in all measured properties: firing rate, directional tuning, and contribution to a decoding model that predicted kinematics from neural activity. The small changes in neural activity that we did observe could be accounted for primarily by subtle changes in behavior. We conclude that the relationship between neural activity and practiced behavior is reasonably stable, at least on timescales of minutes up to 48 h. This finding has significant implications for the design of neural prosthetic systems because it suggests that device recalibration need not be overly frequent, It also has implications for studies of neural plasticity because a stable baseline permits identification of nonstationary shifts. PMID- 17913910 TI - Extracellular signal-regulated kinase 2 (ERK2) knockdown mice show deficits in long-term memory; ERK2 has a specific function in learning and memory. AB - The extracellular signal-regulated kinase (ERK) 1 and 2 are important signaling components implicated in learning and memory. These isoforms display a high degree of sequence homology and share a similar substrate profile. However, recent findings suggest that these isoforms may have distinct roles: whereas ERK1 seems to be not so important for associative learning, ERK2 might be critically involved in learning and memory. Thus, the individual role of ERK2 has received considerable attention, although it is yet to be understood. Here, we have generated a series of mice in which ERK2 expression decreased in an allele dose dependent manner. Null ERK2 knock-out mice were embryonic lethal, and the heterozygous mice were anatomically impaired. To gain a better understanding of the influence of ERK2 on learning and memory, we also generated knockdown mice in which ERK2 expression was partially (20-40%) reduced. These mutant mice were viable and fertile with normal appearance. The mutant mice showed a deficit in long-term memory in classical fear conditioning, whereas short-term memory was normal. The mice also showed learning deficit in the water maze and the eight-arm radial maze. The ERK1 expression level of the knockdown mice was comparable with the wild-type control. Together, our results indicate a noncompensable role of ERK2-dependent signal transduction in learning and memory. PMID- 17913909 TI - Neural correlates of tactile detection: a combined magnetoencephalography and biophysically based computational modeling study. AB - Previous reports conflict as to the role of primary somatosensory neocortex (SI) in tactile detection. We addressed this question in normal human subjects using whole-head magnetoencephalography (MEG) recording. We found that the evoked signal (0-175 ms) showed a prominent equivalent current dipole that localized to the anterior bank of the postcentral gyrus, area 3b of SI. The magnitude and timing of peaks in the SI waveform were stimulus amplitude dependent and predicted perception beginning at approximately 70 ms after stimulus. To make a direct and principled connection between the SI waveform and underlying neural dynamics, we developed a biophysically realistic computational SI model that contained excitatory and inhibitory neurons in supragranular and infragranular layers. The SI evoked response was successfully reproduced from the intracellular currents in pyramidal neurons driven by a sequence of lamina-specific excitatory input, consisting of output from the granular layer (approximately 25 ms), exogenous input to the supragranular layers (approximately 70 ms), and a second wave of granular output (approximately 135 ms). The model also predicted that SI correlates of perception reflect stronger and shorter-latency supragranular and late granular drive during perceived trials. These findings strongly support the view that signatures of tactile detection are present in human SI and are mediated by local neural dynamics induced by lamina-specific synaptic drive. Furthermore, our model provides a biophysically realistic solution to the MEG signal and can predict the electrophysiological correlates of human perception. PMID- 17913912 TI - State-dependent enhancement of subthreshold A-type potassium current by 4 aminopyridine in tuberomammillary nucleus neurons. AB - A-type potassium current (I(A)) both activates and inactivates at subthreshold voltages. We asked whether there is steady-state I(A) at subthreshold voltages, using dissociated mouse tuberomammillary nucleus neurons, pacemaking neurons with large I(A) currents in which subthreshold I(A) might regulate firing frequency. With slow depolarizing voltage ramps (20 mV/s), there was no discernible component of steady-state outward current in the range of -70 to -40 mV. However, faster ramps of 50-100 mV/s, similar to the rate of spontaneous depolarization during pacemaking, did evoke subthreshold outward currents. Ramp-evoked current at subthreshold voltages was unaffected by 10 mM tetraethylammonium and likely represents I(A), because its voltage dependence overlaps that of I(A) activation (midpoint near -44 mV) and inactivation (midpoint near -85 mV). However, although 4-aminopyridine (4-AP) inhibited peak I(A) activated by step depolarizations as expected (IC50, approximately 1 mM), ramp-evoked current was instead dramatically enhanced (current at -40 mV evoked by 50 mV/s ramp enhanced >15-fold by 10 mM 4 AP). In cell-attached recordings of spontaneous pacemaking, 10 mM 4-AP slowed rather than speeded firing, consistent with enhancement of subthreshold I(A). Also consistent with such enhancement, 4-AP also greatly increased the latency to first spike after long hyperpolarizations. The striking enhancement of I(A) during depolarizing ramps can be explained by a model in which 4-AP binds tightly to closed channels but must unbind before channels can inactivate. Thus, the state dependence of 4-AP binding to the channels underlying I(A) can result in effects on firing patterns opposite to those expected from simple block of I(A). PMID- 17913911 TI - Navigating neocortical neurogenesis and neuronal specification: a positional information system encoded by neurogenetic gradients. AB - The projection neurons of the neocortex are produced in the pseudostratified ventricular epithelium (PVE) lining the embryonic lateral ventricles. Over a 7 d period in mouse, these neurons arise in an overlapping layer VI-to-II sequence and in an anterolateral to posteromedial gradient [the transverse neurogenetic gradient (TNG)]. At any time in the 7 d neurogenetic interval, a given PVE cell must know what class of precursor cell or neuron to form next. How this information is encoded in the PVE is not known. With comparative experiments in wild-type and double-transgenic mice, overexpressing the cell cycle inhibitor p27(Kip1), we show that a gradient of expression of Lhx2 (inferred from its mRNA levels), a LIM homeodomain transcription factor, together with a gradient in duration of the G1 phase of the cell cycle (T(G1)), are sufficient to specify a positional mapping system that informs the PVE cell what class of neuron to produce next. Lhx2 likely is representative of an entire class of transcription factors expressed along the TNG. This mapping system consisting of a combination of signals from two different sources is a novel perspective on the source of positional information for neuronal specification in the developing CNS. PMID- 17913914 TI - Paradoxical upregulation of glutamatergic presynaptic boutons during mild cognitive impairment. AB - Synaptic integrity is now recognized as a central component of Alzheimer's disease. Surprisingly, however, the structural status of glutamatergic synapses in Alzheimer's disease is unclear, despite the fact that glutamate is the major excitatory transmitter of the CNS and has key roles in excitotoxicity and long term potentiation. The identification of specific markers of glutamatergic neurons now allows an assessment of the structural involvement of the glutamatergic system across progressive stages of the Alzheimer's pathology, an opportunity not afforded by previously used neurochemical approaches. Glutamatergic presynaptic bouton density and dystrophic neurite abundance were quantified in midfrontal gyrus brain tissue from subjects with no cognitive impairment, mild cognitive impairment, or mild- or severe-stage Alzheimer's disease. Our study demonstrates a striking pathology-dependent pattern of glutamatergic synaptic remodeling with disease progression. Subjects with mild cognitive impairment display a paradoxical elevation in glutamatergic presynaptic bouton density, a situation akin to that observed in the cholinergic system, which then depletes and drops with disease progression. This pattern of synaptic remodeling mirrors our previous findings in transgenic animal models and is of major relevance to current transmitter-based therapeutics. PMID- 17913913 TI - NMDA receptor contribution to the climbing fiber response in the adult mouse Purkinje cell. AB - Among integrative neurons displaying long-term synaptic plasticity, adult Purkinje cells seemed to be an exception by lacking functional NMDA receptors (NMDA-Rs). Although numerous anatomical studies have shown both NR1 and NR2 NMDA R subunits in adult Purkinje cells, patch-clamp studies failed to detect any NMDA currents. Using more recent pharmacological and immunodetection tools, we demonstrate here that Purkinje cells from adult mice respond to exogenous NMDA application and that postsynaptic NMDA-Rs carry part of the climbing fiber mediated EPSC (CF-EPSC), with undetectable contribution from presynaptic or polysynaptic NMDA currents. We also detect NR2-A/B subunits in adult Purkinje cells by immunohistochemistry. The NMDA-mediated CF-EPSC is barely detectable before 3 weeks postnatal. From the end of the third week, the number of cells displaying the NMDA-mediated CF-EPSC rapidly increases. Soon, this EPSC becomes detectable in all the Purkinje cells but is still very small. Its amplitude continues to increase until 12 weeks after birth. In mature Purkinje cells, we show that the NMDA-Rs contribute to the depolarizing plateau of complex spikes and increase their number of spikelets. Together, these observations demonstrate that mature Purkinje cells express functional NMDA receptors that become detectable in CF-EPSCs at approximately 21 d after birth and control the complex spike waveform. PMID- 17913915 TI - State dependence of spike timing and neuronal function in a motor pattern generating network. AB - When sustained firing of a neuron is similar in different types of motor programs, its role in the generation of these programs is often similar. We investigated whether this is also the case for neurons involved in phase transition. In the Aplysia feeding central pattern generator (CPG), identified interneuron B64 starts firing at the transition between the protraction and the retraction phases of all types of motor programs, and its firing is sustained during the retraction phase. It was thought that B64 functions as a protraction terminator as it provides strong inhibitory input to protraction interneurons and motoneurons. Furthermore, premature activation of B64 can lead to premature termination of the protraction phase. Indeed, as we show here, B64 can terminate the protraction phase regardless of the type of motor program. However, B64 actually only functions as a protraction terminator in ingestive-like but not in egestive-like programs. This differential role of B64 results from a differential timing of the initiation of B64 spiking in the two types of programs. In turn, this differential timing of the initiation of B64 firing is determined by the internal state of the CPG. Thus, this study indicates the importance of the timing of initiation of firing in determining the functional role of a neuron and demonstrates that this role depends on the activity-dependent state of the network. PMID- 17913916 TI - Estradiol improves cerebellar memory formation by activating estrogen receptor beta. AB - Learning motor skills is critical for motor abilities such as driving a car or playing piano. The speed at which we learn those skills is subject to many factors. Yet, it is not known to what extent gonadal hormones can affect the achievement of accurate movements in time and space. Here we demonstrate via different lines of evidence that estradiol promotes plasticity in the cerebellar cortex underlying motor learning. First, we show that estradiol enhances induction of long-term potentiation at the parallel fiber to Purkinje cell synapse, whereas it does not affect long-term depression; second, we show that estradiol activation of estrogen receptor beta receptors in Purkinje cells significantly improves gain-decrease adaptation of the vestibulo-ocular reflex, whereas it does not affect general eye movement performance; and third, we show that estradiol increases the density of parallel fiber to Purkinje cell synapses, whereas it does not affect the density of climbing fiber synapses. We conclude that estradiol can improve motor skills by potentiating cerebellar plasticity and synapse formation. These processes may be advantageous during periods of high estradiol levels of the estrous cycle or pregnancy. PMID- 17913917 TI - Breadth of tuning and taste coding in mammalian taste buds. AB - A longstanding question in taste research concerns taste coding and, in particular, how broadly are individual taste bud cells tuned to taste qualities (sweet, bitter, umami, salty, and sour). Taste bud cells express G-protein coupled receptors for sweet, bitter, or umami tastes but not in combination. However, responses to multiple taste qualities have been recorded in individual taste cells. We and others have shown previously there are two classes of taste bud cells directly involved in gustatory signaling: "receptor" (type II) cells that detect and transduce sweet, bitter, and umami compounds, and "presynaptic" (type III) cells. We hypothesize that receptor cells transmit their signals to presynaptic cells. This communication between taste cells could represent a potential convergence of taste information in the taste bud, resulting in taste cells that would respond broadly to multiple taste stimuli. We tested this hypothesis using calcium imaging in a lingual slice preparation. Here, we show that receptor cells are indeed narrowly tuned: 82% responded to only one taste stimulus. In contrast, presynaptic cells are broadly tuned: 83% responded to two or more different taste qualities. Receptor cells responded to bitter, sweet, or umami stimuli but rarely to sour or salty stimuli. Presynaptic cells responded to all taste qualities, including sour and salty. These data further elaborate functional differences between receptor cells and presynaptic cells, provide strong evidence for communication within the taste bud, and resolve the paradox of broad taste cell tuning despite mutually exclusive receptor expression. PMID- 17913918 TI - Moderate reduction of gamma-secretase attenuates amyloid burden and limits mechanism-based liabilities. AB - Although gamma-secretase is recognized as a therapeutic target for Alzheimer's disease, side effects associated with strong inhibition of this aspartyl protease raised serious concerns regarding this therapeutic strategy. However, it is not known whether moderate inhibition of this enzyme will allow dissociation of beneficial effects in the CNS from mechanism-based toxicities in the periphery. We tested this possibility by using a series of mice with genetic reduction of gamma-secretase (levels ranging from 25 to 64% of control mice). Here, we document that even 30% reduction of gamma-secretase can effectively ameliorate amyloid burden in the CNS. However, global reduction of this enzyme below a threshold level increased the risk of developing squamous cell carcinoma as well as abnormal proliferation of granulocytes in a gamma-secretase dosage-dependent manner. Importantly, we demonstrate that there exists a critical gamma-secretase level that reduces the risk of amyloidosis in the CNS and limits tumorigenesis in epithelia. Our findings suggest that moderate inhibition of gamma-secretase represents an attractive anti-amyloid therapy for Alzheimer's disease. PMID- 17913919 TI - Sequential postsynaptic maturation governs the temporal order of GABAergic and glutamatergic synaptogenesis in rat embryonic cultures. AB - Sequential formation of GABAergic and glutamatergic synapses is thought to be crucial for constructing the stereotypic neural networks during brain development. However, why GABAergic synapses are formed earlier than glutamatergic synapses in the developing brain is not well understood. We used electrophysiology and fluorescence imaging to study GABAergic and glutamatergic synaptogenesis in embryonic hypothalamic cultures, which contain approximately 40% GABAergic and approximately 60% glutamatergic neurons. The newly dissociated embryonic hypothalamic neurons contained a significant pool of functional GABA(A) receptors but a very low level of glutamate receptors. Within the first week of culture, the time course of GABAergic synaptogenesis in embryonic neurons coincided with that of presynaptic vesicle cycling, but both measurements lagged behind the detection of functional GABA(A) receptors. Remarkably, the GABA(A) receptors of newly dissociated embryonic neurons can be rapidly clustered into postsynaptic apparatus and generate functional synaptic currents within 4-6 h when cocultured with mature neurons. Consistent with earlier expression of GABA(A) receptors in immature neurons, synaptic GABAergic events were always detected before the onset of glutamatergic events in both purely embryonic and heterochronic cultures. Interestingly, overexpression of glutamate receptors in embryonic neurons not only increased whole-cell glutamate currents but also significantly increased the frequency of excitatory synaptic events. We conclude that the sequential formation of GABAergic and glutamatergic synapses in immature neurons is likely governed by a sequential expression of GABA(A) and glutamate receptors during neuronal development. PMID- 17913921 TI - Serotonin transporter phosphorylation by cGMP-dependent protein kinase is altered by a mutation associated with obsessive compulsive disorder. AB - Human serotonin transporter (hSERT) activity expressed in HeLa cells was stimulated by agents that release nitric oxide, stimulate soluble guanylyl cyclase, or activate cGMP-dependent protein kinase (PKG). This stimulation was blocked by a PKG inhibitor. A naturally occurring mutation, I425V, associated with obsessive-compulsive disorder and other neuropsychiatric disorders, activated hSERT and eliminated stimulation via the PKG pathway. Inhibitors of soluble guanylyl cyclase or PKG decreased activity of the I425V mutant, but not wild type, indicating that both wild-type and mutant transporters could exist in both high and low activity forms. Mutation of Thr-276 in the fifth transmembrane domain (TM5) to alanine or aspartate prevented activation of wild-type hSERT through the PKG pathway and also blocked the inhibition of I425V activity by inhibitors of the pathway. The accessibility of positions in TM5 near Thr-276 was modified in T276D, but not in I425V. These results are consistent with the hypothesis that PKG phosphorylates hSERT at Thr-276 and increases its activity by modifying the substrate permeation pathway formed, in part, by TM5. The effect of the I425V mutation may shift the balance of hSERT toward the phosphorylated form, possibly by interfering with the action of a phosphatase. However, association of hSERT with protein phosphatase 2A was not decreased in the I425V mutant. PMID- 17913920 TI - Hippocampal alpha4beta2 nicotinic acetylcholine receptor involvement in the enhancing effect of acute nicotine on contextual fear conditioning. AB - Nicotine is known to enhance learning and memory in hippocampus-dependent tasks such as contextual fear conditioning. The present study was designed to directly examine whether the hippocampus plays a role in mediating this enhancement and which nicotinic acetylcholine receptor (nAChR) subtypes localized to the hippocampus are critical for enhanced learning. Contextual fear conditioning consisted of two white noise conditioned stimuli presentations, each coterminating with a 2 s, 0.57 mA footshock separated by a 120 s intertrial interval. Nicotine (0.09, 0.18, and 0.35 microg per side) was bilaterally infused into the dorsal hippocampus before training and testing. Infusions of nicotine into the dorsal hippocampus produced a dose-dependent enhancement of contextual fear conditioning. To determine which nAChRs are critical to the enhancing effect of nicotine, the preferential alpha4beta2 nAChR antagonist, dihydro-beta erythroidine (DHbetaE) (6.00 and 18.00 microg per side), or the preferential alpha7 nAChR antagonist, methyllycaconitine (MLA) (13.50 and 27.00 microg per side), was bilaterally infused into the dorsal hippocampus before systemic injections of nicotine (0.09 mg/kg). DHbetaE infusions dose-dependently blocked the enhancement of contextual fear conditioning by nicotine, whereas MLA infusions yielded an intermediate effect. In addition, neither DHbetaE nor MLA had an effect on contextual fear conditioning in the absence of systemic nicotine. The present results suggest a critical role for alpha4beta2 nAChRs in the dorsal hippocampus for mediating the enhancing effect of nicotine on contextual fear conditioning. PMID- 17913922 TI - Bim expression indicates the pathway to retinal cell death in development and degeneration. AB - Programmed cell death (PCD) during development of the mouse retina involves activation of the mitochondrial pathway. Previous work has shown that the multidomain Bcl-2 family proteins Bax and Bak are fundamentally involved in this process. To induce mitochondrial membrane permeabilization, Bax and Bak require that prosurvival members of the family be inactivated by binding of "BH3-only" members. We showed previously that the BH3-only protein BimEL is highly expressed during postnatal retinal development but decreases dramatically thereafter. The purpose of this study was to investigate a possible role for Bim, in retinal development and degeneration, upstream of Bax and Bak. Bim-/- mice analyzed for defective retinal development exhibit an increase in retinal thickness and a delay in PCD, thereby confirming a role for Bim. We also demonstrate that in response to certain death stimuli, bim+/+ retinal explants upregulate BimEL leading to caspase activation and cell death, whereas bim-/- explants are resistant to apoptosis. Finally, we analyzed Bim expression in the retinal degeneration (rd) mouse, an in vivo model of retinal degeneration. Bim isoforms, which decrease during development, are not reexpressed during retinal degeneration and ultimately photoreceptor cells die by a caspase-independent mechanism. Thus, we conclude that in cases in which BimEL is reexpressed during pathological cell death, developmental cell death pathways are reactivated. However, the absence of BimEL expression correlates with caspase-independent death in the rd model. PMID- 17913923 TI - The metalloprotease inhibitor TIMP-3 regulates amyloid precursor protein and apolipoprotein E receptor proteolysis. AB - Cellular cholesterol levels alter the processing of the amyloid precursor protein (APP) to produce Abeta. Activation of liver X receptors (LXRs), one cellular mechanism to regulate cholesterol homeostasis, has been found to alter Abeta levels in vitro and in vivo. To identify genes regulated by LXR, we treated human neuroblastoma cells with an LXR agonist (TO-901317) and examined gene expression by microarray. As expected, TO-901317 upregulated several cholesterol metabolism genes, but it also decreased expression of a metalloprotease inhibitor, TIMP-3. We confirmed this finding using real-time PCR and by measuring TIMP-3 protein in glia, SY5Y cells, and COS7 cells. TIMP-3 is a member of a family of metalloproteinase inhibitors and blocks A disintegrin and metalloproteinase-10 (ADAM-10) and ADAM-17, two APP alpha-secretases. We found that TIMP-3 inhibited alpha-secretase cleavage of APP and an apolipoprotein E (apoE) receptor, ApoER2. TIMP-3 decreased surface levels of ADAM-10, APP, and ApoER2. These changes were accompanied by increased APP beta-C-terminal fragment and Abeta production. These data suggest that TIMP-3 preferentially routes APP and ApoER2 away from the cell surface and alpha-secretase cleavage and encourages endocytosis and beta secretase cleavage. In vivo, TO-901317 decreased brain TIMP-3 levels. TIMP-3 protein levels were increased in human Alzheimer's disease (AD) brain and in APP transgenic mice, suggesting that increased levels of TIMP-3 in AD may contribute to higher levels of Abeta. PMID- 17913924 TI - Distinct modes of neuron addition in adult mouse neurogenesis. AB - Adult neurogenesis is restricted to two distinct areas of the mammalian brain: the olfactory bulb (OB) and the dentate gyrus (DG). Despite its spatial restriction, adult neurogenesis is of crucial importance for sensory processing and learning and memory. Although it has been shown that tens of thousands of new neurons arrive in the OB and DG every day with about half of them surviving after integration, the total contribution of adult neurogenesis to the pre-existing network remains mostly unknown. This is because of previous approaches labeling only a small proportion of adult-generated neurons. Here, we used genetic fate mapping to follow the majority of adult-generated neurons over long periods. Our data demonstrate two distinct modes of neuron addition to the pre-existing network. In the glomerular layer of the OB, there is a constant net addition of adult-generated neurons reaching a third of the total neuronal population within 9 months. In contrast, adult neurogenesis contributes to only a minor fraction of the entire neuronal network in the granular cell layer of the OB and the DG. Although the fraction of adult generated neurons can be further increased by an enriched environment, it still remains a minority of the neuronal network in the DG. Thus, neuron addition is distinct and tightly regulated in the neuronal networks that incorporate new neurons life long. PMID- 17913925 TI - Brain-derived neurotrophic factor expression and respiratory function improve after ampakine treatment in a mouse model of Rett syndrome. AB - Rett syndrome (RTT) is caused by loss-of-function mutations in the gene encoding methyl-CpG-binding protein 2 (MeCP2). Although MeCP2 is thought to act as a transcriptional repressor of brain-derived neurotrophic factor (BDNF), Mecp2 null mice, which develop an RTT-like phenotype, exhibit progressive deficits in BDNF expression. These deficits are particularly significant in the brainstem and nodose cranial sensory ganglia (NGs), structures critical for cardiorespiratory homeostasis, and may be linked to the severe respiratory abnormalities characteristic of RTT. Therefore, the present study used Mecp2 null mice to further define the role of MeCP2 in regulation of BDNF expression and neural function, focusing on NG neurons and respiratory control. We find that mutant neurons express significantly lower levels of BDNF than wild-type cells in vitro, as in vivo, under both depolarizing and nondepolarizing conditions. However, BDNF levels in mutant NG cells can be increased by chronic depolarization in vitro or by treatment of Mecp2 null mice with CX546, an ampakine drug that facilitates activation of glutamatergic AMPA receptors. Ampakine-treated Mecp2 null mice also exhibit marked functional improvement, characterized by restoration of normal breathing frequency and minute volume. These data demonstrate that BDNF expression remains plastic in Mecp2 null mice and raise the possibility that ampakine compounds could be of therapeutic value in the treatment of RTT. PMID- 17913926 TI - Evaluation of the inoculation procedure using a 0.25 McFarland standard for the BD Phoenix automated microbiology system. PMID- 17913927 TI - Multicenter comparison of the VITEK 2 antifungal susceptibility test with the CLSI broth microdilution reference method for testing amphotericin B, flucytosine, and voriconazole against Candida spp. AB - A fully automated commercial antifungal susceptibility test system (VITEK 2; bioMerieux, Inc., Hazelwood, MO) was compared in three different laboratories with the Clinical and Laboratory Standards Institute (formerly the NCCLS) reference broth microdilution method (BMD) by testing 2 quality control strains, 10 reproducibility strains, and 426 isolates of Candida spp. against amphotericin B, flucytosine, and voriconazole. Reference BMD MIC endpoints were established after 24 and 48 h of incubation. VITEK 2 system MIC endpoints were determined spectrophotometrically after 9.1 to 27.1 h of incubation (mean, 12 to 14 h). Excellent essential agreement (within 2 dilutions) between the VITEK 2 system and the 24- and 48-h BMD MICs was observed for all three antifungal agents: amphotericin B, 99.1% and 97%, respectively; flucytosine, 99.1% and 98.8%, respectively; and voriconazole, 96.7% and 96%, respectively. Both intra- and interlaboratory agreements were >98% for all three drugs. The overall categorical agreements between the VITEK 2 system and BMD for flucytosine and voriconazole were 98.1 to 98.6% at the 24-h BMD time point and 96.9 to 97.4% at the 48-h BMD time point. The VITEK 2 system reliably detected flucytosine and voriconazole resistance among Candida spp. and demonstrated excellent quantitative and qualitative agreement with the reference BMD method. PMID- 17913928 TI - Rapid, standardized method for determination of Mycobacterium tuberculosis drug susceptibility by use of mycolic acid analysis. AB - Multidrug-resistant (MDR) Mycobacterium tuberculosis and extrensively drug resistant (XDR) M. tuberculosis are emerging public health threats whose threats are compounded by the fact that current techniques for testing the susceptibility of M. tuberculosis require several days to weeks to complete. We investigated the use of high-performance liquid chromatography (HPLC)-based quantitation of mycolic acids as a means of rapidly determining drug resistance and susceptibility in M. tuberculosis. Standard susceptibility testing and determination of the MICs of drug-susceptible (n = 26) and drug-resistant M. tuberculosis strains, including MDR M. tuberculosis strains (n = 34), were performed by using the Bactec radiometric growth system as the reference method. The HPLC-based susceptibilities of the current first-line drugs, isoniazid (INH), rifampin (RIF), ethambutol (EMB), and pyrazinamide (PZA), were determined. The vials were incubated for 72 h, and aliquots were removed for HPLC analysis by using the Sherlock mycobacterial identification system. HPLC quantitation of total mycolic acid peaks (TMAPs) was performed with treated and untreated cultures. At 72 h, the levels of agreement of the HPLC method with the reference method were 99.5% for INH, EMB, and PZA and 98.7% for RIF. The inter- and intra assay reproducibilities varied by drug, with an average precision of 13.4%. In summary, quantitation of TMAPs is a rapid, sensitive, and accurate method for antibiotic susceptibility testing of all first-line drugs currently used against M. tuberculosis and offers the potential of providing susceptibility testing results within hours, rather than days or weeks, for clinical M. tuberculosis isolates. PMID- 17913929 TI - Emergence of Usutu virus in Hungary. AB - In 2001, Usutu virus (USUV), a mosquito-borne flavivirus of the Japanese encephalitis virus serogroup related to West Nile virus and previously restricted to sub-Saharan Africa, emerged in wild and zoo birds in and around Vienna, Austria. In order to monitor the spread of the infection, a dead bird surveillance program was established in Austria and in neighboring Hungary. In Hungary, 332 dead birds belonging to 52 species were tested for USUV infection between 2003 and 2006. In the first 2 years, all birds investigated were negative. In August 2005, however, USUV was detected in organ samples of a blackbird (Turdus merula), which was found dead in Budapest, Hungary, by reverse transcription-PCR, immunohistochemistry, and in situ hybridization. In July and August 2006, a further six dead blackbirds tested positive for USUV, and the virus was isolated from organ samples of one bird. These birds were also found in urban areas of Budapest. The nearly complete genomic sequence of one Hungarian USUV strain was determined; it was found to share 99.9% identity with the strain that has been circulating in Austria since 2001. This result indicates that the USUV strain responsible for the blackbird die-off in Budapest most likely spread from Austria to Hungary instead of being independently introduced from Africa. PMID- 17913931 TI - Burkholderia pseudomallei endophthalmitis. AB - Melioidosis is an infectious disease caused by Burkholderia pseudomallei, a gram negative bacillus. We report a case of endogenous endophthalmitis caused by B. pseudomallei that was treated with systemic and intravitreal ceftazidime. Finally, the patient achieved a final visual acuity of 20/60 without complication. PMID- 17913930 TI - Mycobacterium avium subspecies paratuberculosis infection in cases of irritable bowel syndrome and comparison with Crohn's disease and Johne's disease: common neural and immune pathogenicities. AB - Mycobacterium avium subsp. paratuberculosis causes Johne's disease, a systemic infection and chronic inflammation of the intestine that affects many species, including primates. Infection is widespread in livestock, and human populations are exposed. Johne's disease is associated with immune dysregulation, with involvement of the enteric nervous system overlapping with features of irritable bowel syndrome in humans. The present study was designed to look for an association between Mycobacterium avium subsp. paratuberculosis infection and irritable bowel syndrome. Mucosal biopsy specimens from the ileum and the ascending and descending colon were obtained from patients with irritable bowel syndrome attending the University of Sassari, Sassari, Sardinia, Italy. Crohn's disease and healthy control groups were also included. Mycobacterium avium subsp. paratuberculosis was detected by IS900 PCR with amplicon sequencing. Data on the potential risk factors for human exposure to these pathogens and on isolates from Sardinian dairy sheep were also obtained. Mycobacterium avium subsp. paratuberculosis was detected in 15 of 20 (75%) patients with irritable bowel syndrome, 3 of 20 (15%) healthy controls, and 20 of 23 (87%) people with Crohn's disease (P = 0.0003 for irritable bowel syndrome patients versus healthy controls and P = 0.0000 for Crohn's disease patients versus healthy controls). One subject in each group had a conserved single-nucleotide polymorphism at position 247 of IS900 that was also found in isolates from seven of eight dairy sheep. There was a significant association (P = 0.0018) between Mycobacterium avium subsp. paratuberculosis infection and the consumption of hand-made cheese. Mycobacterium avium subsp. paratuberculosis is a candidate pathogen in the causation of a proportion of cases of irritable bowel syndrome as well as in Crohn's disease. PMID- 17913932 TI - Molecular typing of a suspected cluster of Nocardia farcinica infections by use of randomly amplified polymorphic DNA, pulsed-field gel electrophoresis, and amplified fragment length polymorphism analyses. AB - Randomly amplified polymorphic DNA (RAPD), pulsed-field gelelectrophoresis (PFGE), and amplified fragment length polymorphism (AFLP) analyses were used to investigate a possible outbreak of Nocardia farcinica. RAPD and PFGE analyses yielded irreproducible and unsatisfactory results, respectively. AFLP analysis seem to be a promising and welcome addition for molecular analysis of Nocardia isolates. PMID- 17913933 TI - Evolution of primary and compensatory lamivudine resistance mutations in chronic hepatitis B virus-infected patients during long-term lamivudine treatment, assessed by a line probe assay. AB - With the availability of more potent nucleotide/nucleoside analogues, the early detection of drug-resistant mutants of hepatitis B virus (HBV) is important for the strategic treatment of chronic hepatitis B. We studied 336 serum samples from 80 patients chronically infected with HBV who were receiving lamivudine treatment for the presence of lamivudine resistance mutations at codons 80, 173, 180, and 204 of the HBV polymerase. The sequencing data were compared with the results generated with the INNO-LiPA HBV DR (drug resistance) v2 strip, a line probe assay (LiPA) covering wild-type and mutant motifs, for resistance mutations to lamivudine and adefovir dipivoxil. This method provided at least the same information as sequencing for 99.1% of all codons analyzed. On the basis of the LiPA results, 20 of 80 patients developed a lamivudine resistance mutation after 1 year. In all 20 patients, the mutation occurred in the YMDD motif at reverse transcriptase position 204 (rt204; M204V/I) either with or without the compensatory mutation at position rt180 (L180M). A compensatory mutation at position rt80 (L80V/I) was detected in half of these patients. After 36 months, a compensatory mutation was seen at position rt173 (V173L) in 3/15 patients. Time to-event survival analysis indicated a 2.8 times greater chance for LiPA to detect a given mutation than sequencing at any moment in time (hazard ratio, 2.8, 95% confidence interval, 1.79, 4.41; P < 0.0001). These results demonstrate that a highly sensitive and specific assay such as the INNO-LiPA HBV DR v2 can precociously detect and monitor the emergence of primary and compensatory lamivudine resistance mutations in patients chronically infected with HBV and is more sensitive than sequencing. PMID- 17913934 TI - Multicenter trials need to use the same assay for hepatitis C virus viral load determination. AB - This study, involving 20 laboratories and using currently available assays for hepatitis C virus RNA quantification, demonstrated that differences in viral load values are due not to interlaboratory variations but rather to the nature of the assay itself. This underlines the importance of using the same assay in multicenter studies or when monitoring antiviral therapy. PMID- 17913935 TI - Suture-related keratitis caused by Corynebacterium macginleyi. AB - We report two cases of suture-related keratitis following penetrating keratoplasty. In both cases, Corynebacterium macginleyi was isolated from corneal specimens. Scanning electron microscopy revealed that corynebacteria could aggregate and form a biofilm. The MICs of sulbenicillin and fluoroquinolones were high for both isolates. Our findings show that C. macginleyi can cause keratitis with biofilm formation. PMID- 17913936 TI - Are oropharyngeal swabs suitable as samples for Legionella-specific PCR testing? PMID- 17913939 TI - p53 and mdm2 expression in colorectal carcinoma: a correlative analysis with clinical staging and histological parameters. AB - Colorectal carcinoma (CRC) is rare in the Indian subcontinent. The two-hit carcinogenic theory has been well established from the turn of this century, in which p53 is described as a late marker in colorectal carcinogenesis, and murine double minute 2 (mdm2) has not yet been correlated with colorectal carcinoma. With the recent introduction of anti-mdm2 oligonucleotides, it is apparent that if its role can be established in colorectal carcinogenesis, the DNA conservation of the wild p53 strain would be achieved therapeutically because mdm2 conserves the wild p53 strain. We studied 32 cases with adequate number of controls and found a positive correlation (P = .001) between these proteins. These proteins were expressed in tumor adjacent to the hyperplastic mucosa, dysplastic mucosa, and aberrant crypt foci, unlike in studies in the West. The expression of these proteins was correlated with aggressive tumor behavior. All these indicate significant diagnostic, therapeutic, and prognostic roles for these proteins. PMID- 17913940 TI - Association of NET-1 gene expression with human hepatocellular carcinoma. AB - NET-1 is a member of the NET-x family. To explore the potential role of NET-1 in hepatocellular carcinoma (HCC), the expression of NET-1 and the relationship with HCC were examined for the first time. We found that NET-1 was frequently expressed in HCC and the peritumor tissue. The relative amounts of NET-1 mRNA in HCC and peritumor tissue were 0.645 +/- 0.37 and 0.466 +/- 0.30, respectively, indicating a higher expression level in HCC than in the peritumor (P < .05). NET 1 protein is usually located on the cell membrane and in the cytoplasm of HCC cells. NET-1 immunoreactivity was found in 126 out of 130 samples of HCC tissue (96.92%). An association of NET-1 expression with cytological variants, histopathological grading, and clinical stages of HCC was also found (P < .05). Detection of NET-1 gene expression in liver biopsy may provide useful information about the biological behavior of HCC. PMID- 17913942 TI - Anatomopathological analysis of sentinel and nonsentinel lymph nodes in breast cancer: hematoxylin-eosin versus immunohistochemistry. AB - The authors compare the detection of metastases in sentinel lymph nodes (SLNs) and nonsentinel lymph nodes (NSLNs) using hematoxylin-eosin (HE) staining versus immunohistochemistry (IHC). Thirty-six patients with breast carcinoma undergo exeresis of the primary tumor and of 50 SLNs and 491 NSLNs. Sentinel lymph nodes are sectioned into transverse slices of 2- to 3-mm thickness, and a cytologic smear and a frozen section were obtained from each slice. The slices are completely cut into serial sections at 100-microm intervals. Two consecutive 4 microm-thick sections are then obtained from each level and were prepared for HE staining and IHC. Nonsentinel lymph nodes are evaluated similarly to SLNs. The authors obtain 4076 SLN sections and 32 012 NSLN sections, for a total of 36 088 sections. A comparison of HE staining versus IHC based on the total number of sections shows a sensitivity of 93.8%, a negative predictive value of 98.9%, and an accuracy of 99.1%. The values obtained by HE staining are similar to those obtained by IHC. PMID- 17913941 TI - Histopathologic diagnosis of celiac disease in children without clinical evidence of malabsorption. AB - Diverse clinical and pathologic experiences seem to have led to the idea that celiac disease is a spectrum in both categories. Conflicting results emerging from different reports have produced a large amount of confusion on the subject. This article discussed histopathology findings in 10 children with positive autoantibodies for celiac disease but without clinical evidence of malabsorption. The patients were evaluated following a detailed video-endoscopic study sampling the proximal (first and second) and distal (third and fourth) duodenal parts separately and processed apart. The procedure consistently revealed advanced villous atrophy in the proximal duodenal mucosa associated with mild to absent involvement of the distal segments. The data here presented favor the interpretation that (1) the presence of autoantibodies for celiac disease is always associated with mucosal damage, (2) mucosal damage in the absence of malabsorption is always evident in the proximal duodenum, and (3) the mucosal biopsy in search for the telltale damage needs to be done in separate samples of proximal and distal duodenal mucosa. This procedure may result in a better understanding of the dynamics of mucosal damage in celiac disease. PMID- 17913943 TI - The theory of the sick breast lobe and the possible consequences. AB - We hypothesize that carcinoma in situ, and consequently breast carcinoma in general, is a lobar disease because the simultaneously or asynchronously appearing, often multiple tumor foci develop within a single lobe. The sick lobe carries some kind of genetic instability already from its initialization during the early embryonic life and is more sensitive to noxious influences than the other lobes within the same breast. Decades of postnatal life with accumulation of additional genetic alterations are needed for malignant transformation of the cells within the sick lobe. The transformation is often multifocal (involving separate distant lobules of this lobe) or diffuse (involving the larger ducts). This hypothesis offers new perspectives in cancer prevention, because selective visualization, excision, or destruction of the sick lobe before development of malignant lesions would substantially reduce the incidence of breast carcinoma. PMID- 17913944 TI - Correlation of clinical, histological, and cytokeratin profiles of squamous cell carcinoma of the oral tongue with prognosis. AB - The aim of the present study was to clinically, morphologically, and immunohistochemically correlate the expression of cytokeratins (CKs) 7, 10, 13, 14, 16, and 19 in 30 cases of tongue squamous cell carcinoma (SCC) with disease outcome, metastases, clinical stage (tumor, node, metastasis [TNM]), and histological grade of malignancy proposed by Bryne. Statistical analysis (chi2 test) showed that only histological grading was not significantly correlated with the clinical variables. CK expression varied in the samples analyzed. CK 10 expression was significantly correlated with the presence of metastases, and the expression of CK 16 was related to disease outcome and also to TNM stages III and IV. These results indicate that metastases and TNM are effective prognostic indicators. The histological grading proposed by Bryne did not reflect the biological behavior of the tongue SCC cases studied. Analysis of some intermediate CK filaments can reflect the biological behavior and aggressiveness of some tongue SCCs. PMID- 17913945 TI - Missed it by that much. PMID- 17913946 TI - Chondroid syringoma with extensive ossification. PMID- 17913947 TI - Signet ring cells in a poorly differentiated Hurthle cell carcinoma of the thyroid combined with two papillary microcarcinomas. PMID- 17913948 TI - Strongyloides hyperinfection in rheumatoid arthritis. PMID- 17913949 TI - Esophageal GIST: case report of surgical enucleation and update on current diagnostic and therapeutic options. AB - Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract, but they have been rarely reported in the esophagus. The authors present the case of an esophageal GIST and discuss the diagnostic course and therapeutic options, as currently reported in the literature. PMID- 17913950 TI - Synchronous gastric gastrointestinal stromal tumor and signet-ring cell adenocarcinoma: a case report. AB - Synchronous tumors of the stomach are uncommon. We present a unique case of gastric synchronous tumors composed of signet-ring cell adenocarcinoma and gastrointestinal stromal tumor (GIST). The two tumors arose at the same site and were sharply juxtaposed without intermingling of morphologically distinct elements. Coincidence probably accounts for this occurrence, even if a common carcinogenic agent had been hypothesized. Preoperative imaging and endoscopic biopsy could lead to the suspicion of synchronous tumors, and an accurate histological identification of both tumors could be achieved by multiple deep endoscopic biopsies. The presence in our case of diffuse carcinomatosis indicates that the signet-ring cell adenocarcinoma had a greater adverse effect on the prognosis than the GIST. PMID- 17913951 TI - Primary malignant melanoma of the transverse colon: report of a case and review of the literature. AB - Gastrointestinal involvement by malignant melanoma is predominantly a metastatic phenomenon. Although primary malignant melanoma of the gastrointestinal tract has been documented in the esophagus, stomach, small bowel, and anorectum, the incidence of primary melanoma of the colon is rare and remains controversial in most cases. We present a case of solitary malignant melanoma of the transverse colon occurring in a 64-year-old African American male patient. Complete dermatologic and ophthalmologic examinations revealed no evidence of a cutaneous or an ocular primary lesion. Microscopic examination of the resection specimen revealed malignant melanoma, which was confirmed by immunohistochemical positivity for S100 and melan-A. We believe that this tumor represents a primary colonic malignant melanoma. PMID- 17913952 TI - Multiple lymphomatous diverticulosis and comorbid chronic lymphocytic leukemia: novel manifestations of ileocolic mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) has tropism for the gastrointestinal tract (GIT) identifiable as multiple polyps and mass lesions throughout the GIT. We describe 2 novel manifestations of MCL. A 60-year-old woman with known chronic lymphocytic leukemia (CLL) had an exophytic mass of the appendiceal orifice. Multiple polypoid masses of the distal ileum were identified in the right hemicolectomy specimen (multiple lymphomatous polyposis). Ancillary studies confirmed the coexistence of the 2 independent lymphoproliferative disorders. A 69-year-old man had recurrent urinary tract infections and pneumatouria caused by a colovesicular fistula complicating diverticulosis coli. Segmental resections of the sigmoid and ileocecum confirmed diverticulosis of the left and right colon. Histology identified infiltrates of MCL confined to the penetrating aspects of colonic diverticula. MCL has not been documented to coexist with CLL. An invaginating morphology of lymphoma, multiple lymphomatous diverticulosis is also a novel presentation. These 2 scenarios expand MCL's known manifestations within the GIT. PMID- 17913953 TI - Cecal adenocarcinoma with prominent rhabdoid feature: report of a case with immunohistochemical, ultrastructural, and molecular analyses. AB - Colorectal adenocarcinoma with rhabdoid phenotype is extremely rare, and only 1 case of adenocarcinoma showing rhabdoid dedifferentiation has been reported. The authors present another case of cecal adenocarcinoma with prominent rhabdoid feature in a 66-year-old man. The 13-cm sized tumor consisted mainly of rhabdoid cells and partly of adenocarcinoma, and transition from adenocarcinoma to rhabdoid areas was noted. Ultrastructural analysis revealed intracytoplasmic aggregates of intermediate filaments in the rhabdoid cells. Adenocarcinoma cells were diffusely immunoreactive to cytokeratin 7 and AE1/3, but occasionally positive for vimentin. The rhabdoid cells were negative for cytokeratin 7, weakly/focally immunoreactive to AE1/3, and diffusely positive for vimentin. These results suggested that the rhabdoid cells were dedifferentiated adenocarcinoma. Analysis of the rhabdoid cells with molecular techniques is also presented. PMID- 17913954 TI - Primary synovial sarcoma of the kidney with rhabdoid features. AB - Synovial sarcoma is a soft tissue sarcoma with clearly defined histologic, immunophenotypic, and molecular features. It occurs predominantly in the extremities of young adults but has been reported in many other anatomic sites. Histologically, it is classified as biphasic, monophasic, and poorly differentiated. The latter category, which includes tumors with a rhabdoid morphology, has been associated with a more aggressive behavior. Generally, the biphasic variant does not pose any diagnostic problem because of its typical histologic appearance; in contrast, the monophasic and poorly differentiated variants may represent a diagnostic challenge because their microscopic features can be confused with those of other spindle cell tumors with rhabdoid features. The application of molecular techniques, such as reverse transcriptase polymerase chain reaction to detect the fusion transcript associated with the characteristic t(X;18) translocation of synovial sarcoma, has enabled the confirmation of this diagnosis, even in cases of unusual localization, such as the one we present here. PMID- 17913955 TI - Primitive small cell tumor with epithelial, gangliocytic, neuroendocrine, and mesenchymal differentiation: report of 2 cases. AB - The authors describe 2 tumors that, to the best of their knowledge, are hitherto undescribed. The predominant cell type was small round to fusiform dark blue cells. The dark blue cells formed distinct epithelial cords with gland-like formations with mucicarmine-positive mucus. Another distinctive component of the tumors was a mesenchymal one. The mesenchymal areas appeared benign and could be likened to a fibroma having a densely collagenous stroma, or they had spindle cells set in the myxoid background, rendering a myxoma-like appearance. Another distinctive feature was ganglion cell differentiation. Mitotic figures, including atypical forms, were found only in the small cell component. All cells were immunohistochemically negative for actin, calponin, desmin, HMB45, neurofilament protein, CD99/MIC2, Melan A, tyrosinase, serotonin, CD56, Melan A, GFAP, and S 100 protein. Cytokeratin, synaptophysin, FLI1 protein, and chromogranin antibodies reacted only in the primitive small round cells, while all the other components were cytokeratin negative. Fluorescence in situ hybridization showed that the tumors are without the EWSR1 gene translocation and gain 12p. Ultrastructurally, the cells were endowed with well-formed intercellular desmosomes membrane-bound secretory in the cytoplasm. Granules were found in the cytoplasm. We suggest the name "primitive small cell tumor with epithelial, gangliocytic, neuroendocrine, and mesenchymal differentiation" for this neoplasm. PMID- 17913956 TI - Ossifying fibromyxoid tumor may express CD56 and CD99: a case report. AB - Ossifying fibromyxoid tumor (OFMT) is an uncommon soft tissue neoplasm characterized by a combination of myxoid and/or fibrous stroma with areas of ossification. Although most authors postulate a neuroectodermal origin for this peculiar tumor, there is no agreement in the literature regarding its histogenesis. In this article, we present the immunohistochemical findings of a case of a 39-year-old white male with an OFMT of the soft tissue in the mandibular region. The tumor was positive to S-100 protein, glial fibrillary acidic protein, CD99, CD56 and negative to smooth muscle actin, cytokeratins AE1/AE3, epithelial membrane antigen, and CD68. To the best of our knowledge, this is the first case reported to be positive to CD56 and CD99. Immunoreactivity to these two antibodies, together with reactivity for S-100 protein and glial fibrillary acidic protein, suggests that OFMT is of a neuroectodermal origin. In our opinion, in the absence of reactivity to at least one neuroectodermal marker one should seriously question a diagnosis of OFMT. PMID- 17913957 TI - Recombination-defective allele uncovers remarkable sex differences. PMID- 17913960 TI - Review article: stem cells in human reproduction. AB - The derivation of human embryonic stem (hES) cells heralds a new era in stem cell research, generating excitement for their therapeutic potential in regenerative medicine. Pioneering work of embryologists, developmental biologists, and reproductive medicine practitioners in in vitro fertilization clinics has facilitated hES cell research. This review summarizes current research focused on optimizing hES cell culture conditions for good manufacturing practice, directing hES cell differentiation toward trophectoderm and germ cells, and approaches used to reprogram cells for pluripotent cell derivation. The identification of germ stem cells in the testis and the recent controversy over their existence in the ovary raise the possibility of harnessing them for treating young cancer survivors. There is also the potential to harvest fetal stem cells with pluripotent cell-like properties from discarded placental tissues. The recent identification of adult stem/progenitor cell activity in the human endometrium offers a new understanding of common gynecological diseases. Discoveries resulting from research into embryonic, germ, fetal, and adult stem cells are highly relevant to human reproduction. PMID- 17913961 TI - A tumor necrosis factor-alpha promoter polymorphism and pregnancy complications: results of a prospective cohort study in 1652 pregnant women. AB - The purpose of this article is to investigate the frequency of the tumor necrosis factor-alpha (TNF-alpha) -308 G/A single nucleotide polymorphism in women with intrauterine fetal death, preeclampsia, preterm delivery, and small-for gestational-age (SGA) infants. In a prospective cohort study, DNA from 1652 consecutive pregnant women was analyzed for TNF-alpha -308 G/A by polymerase chain reaction. Women who developed at least 1 of the predefined pregnancy complications were used as cases and compared to women without pregnancy complications. Of 1652 women, 268 (16.2%) developed at least 1 pregnancy complication. TNF-alpha -308 G/A allele frequencies (G: 463/536 [86%] and A: 73/536 [14%] vs G: 2366/2768 [85%] and A: 402/2768 [15%], respectively; P = .6; odds ratio [OR], 0.93; 95% confidence interval [CI], 0.69-1.25) and genotype distributions (G/G+G/A: 259/268 [97%] and A/A 9/268 [3%] vs G/G+G/A: 1352/1384 [98%] and A/A 32/1384 [2%], respectively; P = .4; OR, 0.20; 95% CI, 0.002-14.81) were not significantly different between cases and controls. The authors observed no statistically significant difference in TNF-a -308 G/A genotype distributions comparing controls and women with intrauterine fetal death, preeclampsia, preterm delivery <34 weeks' gestation, preterm delivery >34 weeks' gestation, SGA infants <3rd percentile, and SGA infants of the 4th to 10th percentile. TNF-alpha -308 G/A is not a genetic marker for identifying women at increased risk of common pregnancy complications. PMID- 17913962 TI - Endothelin-1 attenuates apoptosis in cultured trophoblasts from term human placentas. AB - The authors test the hypothesis that endothelin-1 (ET-1) modulates apoptosis in human term trophoblasts. Primary cultures of cytotrophoblasts from term human placentas (n = 5) were cultured for 16 hours total or 24 hours prior to harvest at 72 hours in atmospheres of <1%, 8%, and 20% oxygen, in the presence of 10% serum, ET-1 (1-100 pmol/mL), both, or neither. The apoptotic cleavage products of poly-ADP-ribose polymerase and cytokeratin 18 filaments were quantified by Western analysis and immunocytochemistry. The expression of BAD, pBAD-serine 112, p53, and 2 isoforms of MDM2 were quantified by immunoblotting, and endothelin A and B receptors were analyzed by immunocytochemistry. Compared to vehicle control, increasing concentrations of ET-1 reduce by 3- to 6-fold the level of apoptosis in cytotrophoblasts exposed to serum-free conditions at 20% oxygen. Similarly, syncytiotrophoblast cultures grown for 24 hours without serum in 100 pmol/mL ET-1 show a 3-fold lower level of apoptosis compared with vehicle control. ET-1 significantly reduces apoptosis in cultures exposed to 20% oxygen but not in cultures exposed to 8% or 1% oxygen. The effect of ET-1 on apoptosis in 20% oxygen is accompanied by reduced p53 expression and is correlated with enhanced expression of endothelin B receptor, compared to cultures in 8% or 1% oxygen. ET-1 reduces apoptosis in cultured human trophoblasts, and this finding suggests a role for ET-1 in protecting trophoblasts against injury. PMID- 17913963 TI - Magnetic resonance signal characteristics of the cervix as pregnancy advances. AB - The objective of this study is to describe magnetic resonance (MR) signal intensity (SI) changes in the cervix during pregnancy. This is an observational cohort study of women with a history of preterm delivery. MR imaging sequences were performed every 3 to 4 weeks. Using 8 regions of interest, the SIs are quantified and analyzed with respect to gestational age. Twenty-seven MR studies were performed on a cohort of 8 women. The SIs of the external os are significantly greater than those of the internal os ( P = .035). Similarly, the SIs of the outer stroma are greater than those of the inner stroma (P = .002). As gestational age advances, the inner to outer stromal SI ratio increases, primarily because of a decreasing SI in the outer stromal layer (P = .03). The MR SIs of the cervical stromal zones display variability during pregnancy and decrease with advancing gestation. PMID- 17913964 TI - The expression of GLUT8, GLUT9a, and GLUT9b in the mouse testis and sperm. AB - The objective of this study is to confirm the precise glucose transporter (GLUT) 8 localization and determine the expression of GLUT9a and GLUT9b by Western blot and confocal and immunoelectron microscopy in the mouse testis and sperm. GLUT8, GLUT9a, and GLUT9b proteins are expressed in the most intraseminiferous tubula cells and Leydig cells. GLUT8 localizes in the midpiece and principal piece as well as in the acrosomal region of the sperm. Immunoelectron microscopic analysis shows that GLUT8 is strongly detectable at the acrosome and neck region of the sperm. In the midpiece, GLUT8 localizes at the outer dense fibers (odf) as well as at the circumference of the spiral mitochondria. In the principal piece, GLUT8 localizes at the odf. GLUT9a strictly localizes in the midpiece, but GLUT9b localizes in the acrosome, midpiece, and principal piece of the sperm. These results suggest that glucose uptake via GLUT8, GLUT9a, and GLUT9b likely affects normal spermatogenesis, steroidogenesis, and sperm function in the mouse. PMID- 17913965 TI - Contractility and calcium signaling of human myometrium are profoundly affected by cholesterol manipulation: implications for labor? AB - The authors elucidate cholesterol's effect on human uterine contractility and calcium signaling to test the hypotheses that elevation of cholesterol decreases uterine activity and that oxytocin cannot augment contraction when cholesterol is elevated. The effects of cholesterol extraction with methyl beta-cyclodextrin and enrichment with low-density lipoproteins and cholesterol on contractile activity and intracellular calcium signaling in spontaneous or oxytocin-stimulated myometrium are determined. Force occurring spontaneously and with oxytocin is significantly increased by cholesterol extraction. Cholesterol enrichment profoundly inhibits force production in a dose-dependent manner and could reverse the effects of cholesterol extraction. Qualitatively similar results are found for nonpregnant and pregnant laboring and non-laboring myometrium. These contractile changes are related to changes in intracellular Ca2+ . Thus, elevated cholesterol is deleterious to contractility and Ca2+ signaling in human myometrium. Cholesterol may contribute to uterine quiescence but could cause difficulties in labor in obese/dyslipidemic women, consistent with their increased cesarean delivery rates. PMID- 17913966 TI - Disordered meiotic regulation of oocytes by duration of diabetes mellitus in BBdp rat. AB - Diabetes mellitus (DM) disturbs normal functions from the level of cells to the level of organs. In this study, the authors explore the detrimental effects of type 1 diabetes on meiotic regulation depending on the duration of DM. In non diabetes-prone BioBreeding (BBdr) control rats, most of the large follicles had germinal vesicle (GV)-intact oocytes. Conversely, a decrease of intact GV that was dependent on the duration of diabetic symptoms was observed; only 54% of the large follicles of diabetes-prone BB (BBdp) rats had GV-intact oocytes at 6 weeks after diabetes induction. Furthermore, some of the secondary follicles in BBdp rats also had germinal vesicle breakdown (GVB) oocytes. The nuclear status of the euglycemia BBdp rat was similar to those of the BBdr rat. In BBdp rats, the rate of meiotic progression to the metaphase II stage was significantly lower; however, the rate of segregated oocytes was significantly increased compared with controls during induction of in vitro maturation. The rate of segregated oocytes was not affected by the presence of the cumulus after chronic symptoms. These results indicate that chronic DM has a detrimental effect on meiotic regulation during folliculogenesis and results in a reduced number of competent oocytes. In addition, these data suggest that the follicle cells can resume supporting the meiotic regulation under euglycemia through insulin administration, independent of the duration of DM. PMID- 17913967 TI - Human nonmetastatic clone 23 type 1 gene suppresses migration of cervical cancer cells and enhances the migration inhibition of fungal immunomodulatory protein from Ganoderma tsugae. AB - The authors investigate the effects of human nonmetastatic clone 23 type 1 (nm23 H1 ) gene and fungal immunomodulatory protein-Ganoderma tsugae (FIP-gts) on the metastatic potential of cervical cancer cells and assess whether nm23-H1 can influence the action of FIP-gts using cell migration and invasion assays and gelatin zymography. The nm23-H1 gene was stably transfected into Caski cells, which lacked nm23-H1 expression. The results show that nm23-H1 stably transfected Caski cells exhibit reduced cell migration but no change of cell invasion and matrix metalloproteinase (MMP)-2 and -9 activities. FIP-gts reduced cell migration in SiHa and nm23-H1 transfected Caski cells more significantly compared with Caski cells and reduced invasion in Caski and nm23-H1-transfected Caski cells, but it exerted no influence on MMP-2 and MMP-9 activities in them. Conclusively, the nm23-H1 gene suppresses cervical cancer cell migration but not invasion and activities of MMP-2 and MMP-9 and enhances the inhibition of FIP-gts upon migration. PMID- 17913968 TI - Relative expression of 1,25-dihydroxyvitamin D3 receptor, vitamin D 1 alpha hydroxylase, vitamin D 24-hydroxylase, and vitamin D 25-hydroxylase in endometriosis and gynecologic cancers. AB - The authors demonstrate expression of the vitamin D receptor (VDR) and its hydroxylases in the endometrium and ovaries of women with and without endometriosis and endometrial or ovarian cancer. Immunohistochemistry showed strong staining of the VDR in endometriosis and endometrial cancer, with the most intense staining in epithelial cells. The VDR mRNA was significantly increased in patients with endometrial and ovarian cancer compared to the control group. There was a significantly higher 1 alpha-hydroxylase expression in the endometrium of patients with endometriosis compared to healthy controls. The observed differences in VDR and 1 alpha -hydroxylase mRNA levels were maintained at the protein level. The authors found no differences in 25-OH vitamin D levels between the serum of patients with endometriosis (25.7 +/- 2.1 ng/mL, n = 46) and healthy controls (22.6 +/- 2.0 ng/mL, n = 33, P = .31). They hypothesize that vitamin D might influence the local activity of immune cells and cytokines thought to play important pathogenic roles in the development and maintenance of endometriosis. PMID- 17913969 TI - Relationships among renal function loss within the normal to mildly impaired range, arterial stiffness, inflammation, and oxidative stress. AB - BACKGROUND AND OBJECTIVES: This study was conducted to clarify whether individuals with mildly impaired renal function show increased arterial stiffness, microinflammation, and oxidative stress as compared with those with normal renal function and also to examine the association of these parameters with the degree of GFR loss in middle-aged Japanese men with a low cardiovascular risk. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: The brachial-ankle pulse wave velocity and plasma levels of C-reactive protein and lipid peroxides were measured in 1873 men (42 +/- 9 yr of age). RESULTS: The brachial-ankle pulse wave velocity but not the plasma C-reactive protein or lipid peroxides, was increased in individuals with mildly impaired renal function. The GFR was significantly correlated with the brachial-ankle pulse wave velocity but not with the log transformed values of C-reactive protein or lipid peroxides. Multivariate linear regression analysis demonstrated a significant relationship between the brachial ankle pulse wave velocity and the GFR, independent of the conventional atherosclerotic risk factors. This relationship was significant even in individuals with GFR values within the "normal renal function" range. Thus, GFR loss seems to be more closely associated with arterial stiffness than with microinflammation and/or oxidative stress. CONCLUSIONS: A weak but significant relationship was observed between the degree of GFR loss and arterial stiffness, even in individuals with GFR values within the normal renal function range. Therefore, increased arterial stiffness may underlie, at least in part, the elevated cardiovascular risk in individuals with mildly impaired renal function. PMID- 17913970 TI - Effects of lanthanum carbonate on the absorption and oral bioavailability of ciprofloxacin. AB - BACKGROUND AND OBJECTIVES: Phosphate binders such as calcium salts or sevelamer, a cationic polymer, can markedly reduce absorption of oral ciprofloxacin. This randomized, open-label, two-way, crossover study examined the influence of the cation lanthanum on systemic ciprofloxacin exposure after oral administration. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Twelve patients randomly received in a crossover manner a single oral dose of ciprofloxacin 750 mg alone and plus lanthanum carbonate 1 g three times daily with meals for six doses, with a washout interval of 7 to 14 d. Serial blood and urine samples were collected for 24 h after ciprofloxacin administration, and ciprofloxacin concentrations were determined using reverse-phase HPLC. Pharmacokinetic parameters of ciprofloxacin were calculated by noncompartmental methods, and the effect of lanthanum on ciprofloxacin pharmacokinetic parameters was assessed using ANOVA. RESULTS: Lanthanum decreased (P < 0.001) the mean ciprofloxacin area under the plasma concentration-time curve by 54% and the maximum plasma concentration by 56%. The 24-h urinary recovery of ciprofloxacin was reduced by 52% by lanthanum (P < 0.001). No statistically significant differences in ciprofloxacin time to maximum plasma concentration, elimination half-life, and renal clearance occurred between the two arms. CONCLUSIONS: Lanthanum carbonate significantly reduces the systemic exposure to orally administered ciprofloxacin. Concomitant administration of both drugs should be avoided to prevent possible suboptimal response to ciprofloxacin. PMID- 17913971 TI - Niacin lowers serum phosphate and increases HDL cholesterol in dialysis patients. AB - BACKGROUND AND OBJECTIVES: Adverse effects complicate the use of drugs that are prescribed for phosphate control in dialysis patients. Alternative treatment options are needed. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: Nicotinic acid inhibits intestinal phosphate reabsorption and increases HDL cholesterol. This study tested the phosphate-lowering and HDL-increasing effect of Niaspan (prolonged-release nicotinic acid) in patients who were undergoing dialysis. Efficacy, safety, and tolerability of Niaspan were prospectively studied. Twenty dialysis patients, who were receiving a stable dosage of a calcium salt containing drug for phosphate control, received after a 2-wk washout period Niaspan for 12 wk. Patients were started on 375 mg/d, and the dosage was increased every 2 wk to achieve 500, 1000, 1500, and 2000 mg/d, respectively. Clinical and laboratory parameters were prospectively recorded in patients who tolerated a target dosage of > or = 1000 mg/d. RESULTS: Seventeen patients tolerated > or = 1000 mg/d Niaspan (mean dosage 1470 +/- 110 mg/d). Niaspan treatment for 12 wk decreased serum phosphate values from 7.2 +/- 0.5 to 5.9 +/- 0.6 mg/dl (P < 0.015). In contrast, Niaspan did not affect serum calcium levels. A significant increase in HDL cholesterol from 40 +/- 3.2 to 59 +/- 5.5 mg/dl (34%) was also observed with Niaspan (P = 0.0005). CONCLUSIONS: Niaspan effectively lowered serum phosphate levels and significantly increased HDL cholesterol. Niaspan may provide an alternative or adjunctive treatment option in dialysis patients. PMID- 17913972 TI - Hypertonic saline for hyponatremia: risk of inadvertent overcorrection. AB - BACKGROUND AND OBJECTIVES: Data regarding dosage-response relationships for using hypertonic saline in treatment of hyponatremia are extremely limited. Objectives of this study were to assess adherence to previously published guidelines (limiting correction to <12 mEq/L per d and <18 mEq/L per 48 h) in treating hyponatremia with hypertonic saline and to determine the predictive accuracy of the Adrogue-Madias formula. DESIGN, SETTING, PARTICIPANTS & MEASUREMENTS: A retrospective review was conducted of all 62 adult, hyponatremic patients who were treated with hypertonic saline during 5 yr at a 528-bed, acute care, teaching hospital. RESULTS: Median infusion rate was 0.38 ml/kg per h, increasing serum sodium concentration by 0.47 +/- 0.05 mEq/L per h, 7.1 +/- 0.6 mEq/L per 24 h, and 11.3 +/- 0.7 mEq/L per 48 h. In 11.3% of cases, the increase was >12 mEq/L per 24 h and in 9.7% was >18 mEq/L per 48 h. No patient's rate was corrected by >25 mEq/L per 48 h. Among patients with serum sodium <120 mEq/L, the observed increase in sodium exceeded the rise predicted by the Adrogue-Madias formula in 74.2%; the average correction in overcorrectors was 2.4 times the predicted. Inadvertent overcorrection was due to documented water diuresis in 40% of cases. CONCLUSIONS: The Adrogue-Madias formula underestimates increase in sodium concentration after hypertonic saline therapy. Unrecognized hypovolemia and other reversible causes of water retention pose a risk for inadvertent overcorrection. Hypertonic saline should be infused at rates lower than those predicted by formulas with close monitoring of serum sodium and urine output. PMID- 17913973 TI - Bovine monocyte TLR2 receptors differentially regulate the intracellular fate of Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium. AB - Pathogenic mycobacterial organisms have the capacity to inhibit macrophage activation and phagosome maturation. Although the mechanism is complex, several studies have incriminated signaling through TLR2 receptors with subsequent activation of the MAPK pathway p38 (MAPKp38) and overproduction of IL-10 in the survival of pathogenic mycobacterial organisms. In the present study, we compared the response of bovine monocytes with infection by Mycobacterium avium subspecies paratuberculosis (MAP), the cause of paratuberculosis in ruminants, with the closely related organism M. avium subspecies avium (Maa), which usually does not cause disease in ruminants. Both MAP and Maa induced phosphorylation of MAPKp38 by bovine monocytes; however, addition of a blocking anti-TLR2 antibody partially prevented MAPKp38 phosphorylation of MAP-infected monocytes but not Maa-infected monocytes. Addition of anti-TLR2 antibody enhanced phagosome acidification and phagosome-lysosome fusion in MAP-containing phagosomes and enabled monocytes to kill MAP organisms. These changes were not observed in Maa-infected monocytes. The effect on phagosome maturation appears to occur independently from the previously described inhibitory effects of IL-10 on phagosome acidification and organism killing, as IL-10 production was not affected by addition of anti-TLR2 antibody to monocyte cultures. Therefore, signaling through the TLR2 receptor appears to play a role in phagosome trafficking and antimicrobial responses in MAP-infected bovine mononuclear phagocytes. PMID- 17913974 TI - Regulatory T cell populations in sepsis and trauma. AB - Sepsis syndrome remains the leading cause of mortality in intensive care units. It is now believed that along with the body's hyperinflammatory response designated to eliminate the underlying pathogen, mechanisms are initiated to control this initial response, which can become deleterious and result in immune dysfunctions and death. A similar state of immune suppression has been described after numerous forms of severe trauma/injury. Although the evidence for immune dysfunctions after sepsis has grown, much remains to be understood about mechanisms underpinning its development and how it acts to increase the morbid state of the critically ill patient. In this context, although the majority of clinical and basic science conducted so far has focused on the roles of myeloid cell populations, the contribution of T lymphocytes and in particular, of regulatory T cells has been somewhat ignored. The studies presented here support the concept that regulatory T lymphocytes (CD4+CD25+ regulatory, gammadelta, and NK T cells) play a role in the control of immune responses and are affected by injury and sepsis. This may be related to their capacity to interact with components of the innate and adaptive immune responses and to their ability to be activated nonspecifically by bacterial products and/or cytokines and to regulate through direct cell-cell and/or soluble mediators. It is our hope that a better understanding of the mechanism through which those rare lymphocyte subsets exert such a profound effect on the immune response may help in improving our ability not only to diagnose but also to treat the critically ill individual. PMID- 17913975 TI - Pivotal advance: inhibition of HMGB1 nuclear translocation as a mechanism for the anti-rheumatic effects of gold sodium thiomalate. AB - Gold compounds such as gold sodium thiomalate (GST) can reduce the symptoms of rheumatoid arthritis (RA), although their mechanism of action is not well defined. As the proinflammatory mediator high mobility group box chromosomal protein 1 (HMGB1) may play a role in the pathogenesis of RA, we have performed in vitro studies to investigate whether GST inhibits HMGB1 release as the basis of its mode of action. Murine RAW 264.7 or human THP-1 macrophage cells were stimulated in culture with agents causing extracellular HMGB1 release, including LPS, IFN-gamma, polyinosinic:polycytidylic acid, IFN-beta, or NO in the presence of GST, ranging from 0 microM to 250 microM. Secretion and intracellular location of HMGB1 were assessed by Western blotting, HMGB1-specific ELISPOT assay, and immunofluorescent staining. In parallel, TNF and IFN-beta levels were analyzed by ELISPOT and/or ELISA. Supernatant NO production was analyzed by the Griess method. At pharmacologically relevant doses, GST inhibited the extracellular release of HMGB1 from activated macrophages and caused the nuclear retention of this protein; in contrast, no effects were observed on the secretion or production of TNF. Release of the key endogenous mediators of HMGB1 translocation, IFN-beta and NO, was inhibited by GST. This inhibition required gold, as sodium thiomalate did not affect the responses measured. Furthermore, gold chloride also inhibited release of HMGB1. Together, these results suggest a new mechanism for the anti-rheumatic effects of gold salts in RA and the potential of drugs, which interfere with intracellular HMGB1 transport mechanisms, as novel agents to treat RA. PMID- 17913976 TI - Involvement of LTB4 in zymosan-induced joint nociception in mice: participation of neutrophils and PGE2. AB - Leukotriene B4 (LTB4) mediates different inflammatory events such as neutrophil migration and pain. The present study addressed the mechanisms of LTB4-mediated joint inflammation-induced hypernociception. It was observed that zymosan-induced articular hypernociception and neutrophil migration were reduced dose-dependently by the pretreatment with MK886 (1-9 mg/kg; LT synthesis inhibitor) as well as in 5-lypoxygenase-deficient mice (5LO(-/-)) or by the selective antagonist of the LTB(4) receptor (CP105696; 3 mg/kg). Histological analysis showed reduced zymosan induced articular inflammatory damage in 5LO(-/-) mice. The hypernociceptive role of LTB4 was confirmed further by the demonstration that joint injection of LTB4 induces a dose (8.3, 25, and 75 ng)-dependent articular hypernociception. Furthermore, zymosan induced an increase in joint LTB4 production. Investigating the mechanism underlying LTB4 mediation of zymosan-induced hypernociception, LTB4 induced hypernociception was reduced by indomethacin (5 mg/kg), MK886 (3 mg/kg), celecoxib (10 mg/kg), antineutrophil antibody (100 mug, two doses), and fucoidan (20 mg/kg) treatments as well as in 5LO(-/-) mice. The production of LTB4 induced by zymosan in the joint was reduced by the pretreatment with fucoidan or antineutrophil antibody as well as the production of PGE2 induced by LTB4. Therefore, besides reinforcing the role of endogenous LTB4 as an important mediator of inflamed joint hypernociception, these results also suggested that the mechanism of LTB4-induced articular hypernociception depends on prostanoid and neutrophil recruitment. Furthermore, the results also demonstrated clearly that LTB4-induced hypernociception depends on the additional release of endogenous LTs. Concluding, targeting LTB4 synthesis/action might constitute useful therapeutic approaches to inhibit articular inflammatory hypernociception. PMID- 17913978 TI - Control of movement variability and the regulation of limb impedance. AB - Humans routinely make movements to targets that have different accuracy requirements in different directions. Examples extend from everyday occurrences such as grasping the handle of a coffee cup to the more refined instance of a surgeon positioning a scalpel. The attainment of accuracy in situations such as these might be related to the nervous system's capacity to regulate the limb's resistance to displacement, or impedance. To test this idea, subjects made movements from random starting locations to targets that had shape-dependent accuracy requirements. We used a robotic device to assess both limb impedance and patterns of movement variability just as the subject reached the target. We show that impedance increases in directions where required accuracy is high. Independent of target shape, patterns of limb stiffness are seen to predict spatial patterns of movement variability. The nervous system is thus seen to modulate limb impedance in entirely predictable environments to aid in the attainment of reaching accuracy. PMID- 17913977 TI - Activity of murine raphe magnus cells predicts tachypnea and on-going nociceptive responsiveness. AB - In rats, opioids produce analgesia in large part by their effects on two cell populations in the medullary raphe magnus (RM). To extend our mechanistic understanding of opioid analgesia to the genetically tractable mouse, we characterized behavioral reactions and RM neural responses to opioid administration. d-Ala(2), N-Me-Phe(4)-Gly(5)ol-enkephalin, a mu-opioid receptor agonist, microinjected into the murine RM produced cardiorespiratory depression and reduced slow wave electroencephalographic activity as well as increased the noxious heat-evoked withdrawal latencies. As in rat, RM cell types that were excited and inhibited by noxious stimuli, termed on and off cells, respectively, were observed in mice. However, in contrast to findings in rat, opioid doses that suppressed withdrawals did not alter the background discharge rate of murine on and off cells, suggesting that the cellular mechanisms by which the murine RM generates opioid analgesia are substantially different from those in rats. Murine on cell discharge did not predict the latency or magnitude of an ensuing withdrawal but did correlate to the magnitude and latency of concurrent withdrawals. Although opioids failed to alter the background discharge of on and off cells, they reduced the responses of RM neurons to noxious stimulation, further evidence that RM modulates on-going withdrawals. In characterizing the role of RM in respiratory modulation, we found that on cells burst and off cells paused during tachypneic events. The effects of opioids in the murine RM on homeostasis and the association of on and off cell discharge with tachypnea corroborate roles for opioid signaling in RM beyond analgesia. PMID- 17913980 TI - Effects of dopaminergic modulation on the integrative properties of the ventral striatal medium spiny neuron. AB - Dopaminergic modulation produces a variety of functional changes in the principal cell of the striatum, the medium spiny neuron (MSN). Using a 189-compartment computational model of a ventral striatal MSN, we simulated whole cell D1- and D2 receptor-mediated modulation of both intrinsic (sodium, calcium, and potassium) and synaptic currents (AMPA and NMDA). Dopamine (DA) modulations in the model were based on a review of published experiments in both ventral and dorsal striatum. To objectively assess the net effects of DA modulation, we combined reported individual channel modulations into either D1- or D2-receptor modulation conditions and studied them separately. Contrary to previous suggestions, we found that D1 modulation had no effect on MSN nonlinearity and could not induce bistability. In agreement with previous suggestions, we found that dopaminergic modulation leads to changes in input filtering and neuronal excitability. Importantly, the changes in neuronal excitability agree with the classical model of basal ganglia function. We also found that DA modulation can alter the integration time window of the MSN. Interestingly, the effects of DA modulation of synaptic properties opposed the effects of DA modulation of intrinsic properties, with the synaptic modulations generally dominating the net effect. We interpret this lack of synergy to suggest that the regulation of whole cell integrative properties is not the primary functional purpose of DA. We suggest that D1 modulation might instead primarily regulate calcium influx to dendritic spines through NMDA and L-type calcium channels, by both direct and indirect mechanisms. PMID- 17913981 TI - Effects of initial eye position on saccade-related behavior of abducens nucleus neurons in the primate. AB - Previous work suggests that when the eye starts at different orbital initial positions (IPs), the saccade control system is faced with significant nonlinearities. Here we studied the effects of IP on saccade-related firing of monkey abducens neurons by either isolating saccade variables behaviorally or applying a multiple linear regression analysis. Over a 50 degrees range of IPs, we could select 10 degrees horizontal saccades with identical velocity profiles, which would require identical control signals in a linear system. The bursts accompanying ipsiversive saccades for IPs above the threshold for steady firing were quite similar. The excess burst rate above steady firing was either constant or decreased with ipsiversive IP, and both the number of excess spikes in the burst and burst duration were nearly constant. However, for ipsiversive saccades from IPs below threshold, both peak burst rate (6.82 +/- 1.38 spikes.s(-1).deg( 1)) and burst duration (0.67 +/- 0.28 ms/deg) increased substantially with ipsiversive IPs. Moreover, the pause associated with contraversive saccades shortened considerably with ipsiversive IPs (mean 1.2 ms/deg). This pattern of results for pauses and for bursts below threshold suggests the presence of a significant nonlinearity. Abducting saccades are produced by the net force of agonist lateral rectus (LR) and antagonist medial rectus (MR) muscles. We suggest that the decreasing force in the MR muscle with IPs in the abducting direction requires a more vigorous burst in LR motoneurons, which appears to be generated by a combination of saturating and nonsaturating burst commands and the recruitment of additional abducens neurons. PMID- 17913979 TI - The hemo-neural hypothesis: on the role of blood flow in information processing. AB - Brain vasculature is a complex and interconnected network under tight regulatory control that exists in intimate communication with neurons and glia. Typically, hemodynamics are considered to exclusively serve as a metabolic support system. In contrast to this canonical view, we propose that hemodynamics also play a role in information processing through modulation of neural activity. Functional hyperemia, the basis of the functional MRI (fMRI) BOLD signal, is a localized influx of blood correlated with neural activity levels. Functional hyperemia is considered by many to be excessive from a metabolic standpoint, but may be appropriate if interpreted as having an activity-dependent neuro-modulatory function. Hemodynamics may impact neural activity through direct and indirect mechanisms. Direct mechanisms include delivery of diffusible blood-borne messengers and mechanical and thermal modulation of neural activity. Indirect mechanisms are proposed to act through hemodynamic modulation of astrocytes, which can in turn regulate neural activity. These hemo-neural mechanisms should alter the information processing capacity of active local neural networks. Here, we focus on analysis of neocortical sensory processing. We predict that hemodynamics alter the gain of local cortical circuits, modulating the detection and discrimination of sensory stimuli. This novel view of information processing that includes hemodynamics as an active and significant participant-has implications for understanding neural representation and the construction of accurate brain models. There are also potential medical benefits of an improved understanding of the role of hemodynamics in neural processing, as it directly bears on interpretation of and potential treatment for stroke, dementia, and epilepsy. PMID- 17913983 TI - Synchronization measurement of multiple neuronal populations. AB - The purpose of the present paper is to develop a method, based on equal-time correlation, correlation matrix analysis and surrogate resampling, that is able to quantify and describe properties of synchronization of population neuronal activity recorded simultaneously from multiple sites. Initially, Lorenz-type oscillators were used to model multiple time series with different patterns of synchronization. Eigenvalue and eigenvector decomposition was then applied to identify "clusters" of locally synchronized activity and to calculate a "global synchronization index." This method was then applied to multichannel data recorded from an in vitro model of epileptic seizures. The results demonstrate that this novel method can be successfully used to analyze synchronization between multiple neuronal population series. PMID- 17913984 TI - Feeding CPG in Aplysia directly controls two distinct outputs of a compartmentalized interneuron that functions as a CPG element. AB - In the context of motor program generation in Aplysia, we characterize several functional aspects of intraneuronal compartmentalization in an interganglionic interneuron, CBI-5/6. CBI-5/6 was shown previously to have a cerebral compartment (CC) that includes a soma that does not generate full-size action potentials and a buccal compartment (BC) that does. We find that the synaptic connections made by the BC of CBI-5/6 in the buccal ganglion counter the activity of protraction phase neurons and reinforce the activity of retraction-phase neurons. In buccal motor programs, the BC of CBI-5/6 fires phasically, and its premature activation can phase advance protraction termination and retraction initiation. Thus the BC of CBI-5/6 can act as an element of the central pattern generator (CPG). During protraction, the CC of CBI-5/6 receives direct excitatory inputs from the CPG elements, B34 and B63, and during retraction, it receives antidromically propagating action potentials that originate in the BC of CBI-5/6. Consequently, in its CC, CBI-5/6 receives depolarizing inputs during both protraction and retraction, and these depolarizations can be transmitted via electrical coupling to other neurons. In contrast, in its BC, CBI-5/6 uses spike-dependent synaptic transmission. Thus the CPG directly and differentially controls the program phases in which the two compartments of CBI-5/6 may transmit information to its targets. PMID- 17913982 TI - Spatial and functional architecture of the mammalian brain stem respiratory network: a hierarchy of three oscillatory mechanisms. AB - Mammalian central pattern generators (CPGs) producing rhythmic movements exhibit extremely robust and flexible behavior. Network architectures that enable these features are not well understood. Here we studied organization of the brain stem respiratory CPG. By sequential rostral to caudal transections through the pontine medullary respiratory network within an in situ perfused rat brain stem-spinal cord preparation, we showed that network dynamics reorganized and new rhythmogenic mechanisms emerged. The normal three-phase respiratory rhythm transformed to a two-phase and then to a one-phase rhythm as the network was reduced. Expression of the three-phase rhythm required the presence of the pons, generation of the two-phase rhythm depended on the integrity of Botzinger and pre Botzinger complexes and interactions between them, and the one-phase rhythm was generated within the pre-Botzinger complex. Transformation from the three-phase to a two-phase pattern also occurred in intact preparations when chloride mediated synaptic inhibition was reduced. In contrast to the three-phase and two phase rhythms, the one-phase rhythm was abolished by blockade of persistent sodium current (I(NaP)). A model of the respiratory network was developed to reproduce and explain these observations. The model incorporated interacting populations of respiratory neurons within spatially organized brain stem compartments. Our simulations reproduced the respiratory patterns recorded from intact and sequentially reduced preparations. Our results suggest that the three phase and two-phase rhythms involve inhibitory network interactions, whereas the one-phase rhythm depends on I(NaP). We conclude that the respiratory network has rhythmogenic capabilities at multiple levels of network organization, allowing expression of motor patterns specific for various physiological and pathophysiological respiratory behaviors. PMID- 17913986 TI - Relationship of semicircular canal size to vestibular-nerve afferent sensitivity in mammals. AB - The relationship between semicircular canal radius of curvature and afferent sensitivity has not been experimentally determined. We characterized mouse semicircular canal afferent responses to sinusoidal head rotations to facilitate interspecies and intraspecies comparisons of canal size to sensitivity. The interspecies experiment compared the horizontal canal afferent responses among animals ranging in size from mouse to rhesus monkey. The intraspecies experiment compared afferent responses from the larger anterior canal to those from the smaller horizontal canal of mice. The responses of mouse vestibular-nerve afferents showed a low- and high-frequency phase lead and high-frequency gain enhancement. Regular horizontal-canal afferents showed a sensitivity to 0.5-Hz sinusoidal rotations of 0.10 +/- 0.03 (SD) spike . s(-1)/deg . s(-1) and high gain irregular afferents showed a sensitivity of 0.25 +/- 0.11 spike . s(-1)/deg . s(-1). The interspecies comparison showed that the sensitivity of regular afferents was related to the radius of curvature R according to the formula G(r) = 0.23R - 0.09 (r(2) = 0.86) and the sensitivity of irregular afferents was related to radius according to the formula G(i) = 0.32R + 0.01 (r(2) = 0.67). The intraspecies comparison showed that regularly firing anterior canal afferents were significantly more sensitive than those from the relatively smaller horizontal canal, with G(r) = 0.25R. This suggests that canal radius of curvature is closely related to afferent sensitivity both among and within species. If the relationship in humans is similar to that demonstrated here, the sensitivity of their regular vestibular-nerve afferents to 0.5-Hz rotations is likely to be about 0.67 spike . s(-1)/deg . s(-1) and of their high-gain irregular afferents about 1.06 spikes . s(-1)/deg . s(-1). PMID- 17913985 TI - Variability in responses and temporal coding of tastants of similar quality in the nucleus of the solitary tract of the rat. AB - In the nucleus of the solitary tract (NTS), electrophysiological responses to taste stimuli representing four basic taste qualities (sweet, sour, salty, or bitter) can often be discriminated by spike count, although in units for which the number of spikes is variable across identical stimulus presentations, spike timing (i.e., temporal coding) can also support reliable discrimination. The present study examined the contribution of spike count and spike timing to the discrimination of stimuli that evoke the same taste quality but are of different chemical composition. Responses to between 3 and 21 repeated presentations of two pairs of quality-matched tastants were recorded from 38 single cells in the NTS of urethane-anesthetized rats. Temporal coding was assessed in 24 cells, most of which were tested with salty and sour tastants, using an information-theoretic approach. Within a given cell, responses to tastants of similar quality were generally closer in magnitude than responses to dissimilar tastants; however, tastants of similar quality often reversed their order of effectiveness across replicate sets of trials. Typically, discrimination between tastants of dissimilar qualities could be made by spike count. Responses to tastants of similar quality typically evoked more similar response magnitudes but were more frequently, and to a proportionally greater degree, distinguishable based on temporal information. Results showed that nearly every taste-responsive NTS cell has the capacity to generate temporal features in evoked spike trains that can be used to distinguish between stimuli of different qualities and chemical compositions. PMID- 17913987 TI - Peptide hormone modulation of a neuronally modulated motor circuit. AB - Rhythmically active motor circuits are influenced by neuronally released and circulating hormone modulators, but there are few systems in which the influence of a peptide hormone modulator on a neuronally modulated motor circuit has been determined. We performed such an analysis in the isolated crab stomatogastric nervous system by assessing the influence of the hormone crustacean cardioactive peptide (CCAP) on the gastric mill (chewing) rhythm elicited by identified modulatory projection neurons. The gastric mill circuit is located in the stomatogastric ganglion. In situ, this ganglion is located within the ophthalmic artery and thus is in the path of circulating hormones such as CCAP. Focally applied CCAP directly excited some gastric mill neurons, including the gastric mill central pattern generator neurons LG and Int1, but it did not elicit a sustained gastric mill rhythm. At concentrations as low as 10(-10) M, however, CCAP did influence gastric mill rhythms elicited by coactivating the projection neurons MCN1 and CPN2 and by selectively stimulating MCN1. In both cases, CCAP slowed this rhythm by selectively prolonging the protraction phase, although its influence on the MCN1-elicited rhythm was limited to those with relatively brief cycle periods. Interestingly, CCAP also reduced the threshold MCN1 firing frequency for activating the gastric mill rhythm. Last, the gastric mill neurons that exhibited altered activity during these CCAP-influenced rhythms did not correspond completely to the set of CCAP-responsive neurons. These results highlight the ability of hormonal modulation to enhance the flexibility provided by the neuronal modulation of rhythmically active motor circuits. PMID- 17913988 TI - Dependence of response properties on sparse connectivity in a spiking neuron model of the lateral geniculate nucleus. AB - We present a large-scale anatomically constrained spiking neuron model of the lateral geniculate nucleus (LGN), which operates solely with retinal input, relay cells, and interneurons. We show that interneuron inhibition and sparse connectivity between LGN cells could be key factors for explaining a number of observed classical and extraclassical response properties in LGN of monkey and cat. Among them are 1) weak orientation tuning, 2) contrast invariance of spatial frequency tuning in the absence of cortical feedback, 3) extraclassical surround suppression, and 4) orientation tuning of extraclassical surround suppression. The model also makes two surprising predictions: 1) a possible pinwheel-like spatial organization of orientation preference in the parvo layers of monkey LGN, much like what is seen in V1, and 2) a stimulus-induced trend (bias) in the orientation and phase preference of surround suppression, originating from the stimulus discontinuity between center and surround gratings rather than from specific circuitry. PMID- 17913989 TI - Sustained rhythmic activity in gap-junctionally coupled networks of model neurons depends on the diameter of coupled dendrites. AB - Gap junctions are known to be important for many network functions such as synchronization of activity and the generation of waves and oscillations. Gap junctions have also been proposed to be essential for the generation of early embryonic activity. We have previously shown that the amplitude of electrical signals propagating across gap-junctionally coupled passive cables is maximized at a unique diameter. This suggests that threshold-dependent signals may propagate through gap junctions for a finite range of diameters around this optimal value. Here we examine the diameter dependence of action potential propagation across model networks of dendro-dendritically coupled neurons. The neurons in these models have passive soma and dendrites and an action potential generating axon. We show that propagation of action potentials across gap junctions occurs only over a finite range of dendritic diameters and that propagation delay depends on this diameter. Additionally, in networks of gap junctionally coupled neurons, rhythmic activity can emerge when closed loops (re entrant paths) occur but again only for a finite range of dendrite diameters. The frequency of such rhythmic activity depends on the length of the path and the dendrite diameter. For large networks of randomly coupled neurons, we find that the re-entrant paths that underlie rhythmic activity also depend on dendrite diameter. These results underline the potential importance of dendrite diameter as a determinant of network activity in gap-junctionally coupled networks, such as network rhythms that are observed during early nervous system development. PMID- 17913990 TI - Synapsin regulates Basal synaptic strength, synaptic depression, and serotonin induced facilitation of sensorimotor synapses in Aplysia. AB - Synapsin is a synaptic vesicle-associated protein implicated in the regulation of vesicle trafficking and transmitter release, but its role in heterosynaptic plasticity remains elusive. Moreover, contradictory results have obscured the contribution of synapsin to homosynaptic plasticity. We previously reported that the neuromodulator serotonin (5-HT) led to the phosphorylation and redistribution of Aplysia synapsin, suggesting that synapsin may be a good candidate for the regulation of vesicle mobilization underlying the short-term synaptic plasticity induced by 5-HT. This study examined the role of synapsin in homosynaptic and heterosynaptic plasticity. Overexpression of synapsin reduced basal transmission and enhanced homosynaptic depression. Although synapsin did not affect spontaneous recovery from depression, it potentiated 5-HT-induced dedepression. Computational analysis showed that the effects of synapsin on plasticity could be adequately simulated by altering the rate of Ca(2+)-dependent vesicle mobilization, supporting the involvement of synapsin not only in homosynaptic but also in heterosynaptic forms of plasticity by regulating vesicle mobilization. PMID- 17913991 TI - Responses of inferior colliculus neurons to double harmonic tones. AB - The auditory system can segregate sounds that overlap in time and frequency, if the sounds differ in acoustic properties such as fundamental frequency (f0). However, the neural mechanisms that underlie this ability are poorly understood. Responses of neurons in the inferior colliculus (IC) of the anesthetized chinchilla were measured. The stimuli were harmonic tones, presented alone (single harmonic tones) and in the presence of a second harmonic tone with a different f0 (double harmonic tones). Responses to single harmonic tones exhibited no stimulus-related temporal pattern, or in some cases, a simple envelope modulated at f0. Responses to double harmonic tones exhibited complex slowly modulated discharge patterns. The discharge pattern varied with the difference in f0 and with characteristic frequency. The discharge pattern also varied with the relative levels of the two tones; complex temporal patterns were observed when levels were equal, but as the level difference increased, the discharge pattern reverted to that associated with single harmonic tones. The results indicated that IC neurons convey information about simultaneous sounds in their temporal discharge patterns and that the patterns are produced by interactions between adjacent components in the spectrum. The representation is "low-resolution," in that it does not convey information about single resolved components from either individual sound. PMID- 17913992 TI - Zinc enhances the inhibitory effects of strychnine-sensitive glycine receptors in mouse hippocampal neurons. AB - Although extracellular Zn(2+) is an endogenous biphasic modulator of strychnine sensitive glycine receptors (GlyRs), the physiological significance of this modulation remains poorly understood. Zn(2+) modulation of GlyR may be especially important in the hippocampus where presynaptic Zn(2+) is abundant. Using cultured embryonic mouse hippocampal neurons, we examined whether 1 microM Zn(2+), a potentiating concentration, enhances the inhibitory effects of GlyRs activated by sustained glycine applications. Sustained 20 microM glycine (EC(25)) applications alone did not decrease the number of action potentials evoked by depolarizing steps, but they did in 1 microM Zn(2+). At least part of this effect resulted from Zn(2+) enhancing the GlyR-induced decrease in input resistance. Sustained 20 microM glycine applications alone did not alter neuronal bursting, a form of hyperexcitability induced by omitting extracellular Mg(2+). However, sustained 20 microM glycine applications depressed neuronal bursting in 1 microM Zn(2+). Zn(2+) did not enhance the inhibitory effects of sustained 60 microM glycine (EC(70)) applications in these paradigms. These results suggest that tonic GlyR activation could decrease neuronal excitability. To test this possibility, we examined the effect of the GlyR antagonist strychnine and the Zn(2+) chelator tricine on action potential firing by CA1 pyramidal neurons in mouse hippocampal slices. Co-applying strychnine and tricine slightly but significantly increased the number of action potentials fired during a depolarizing current step and decreased the rheobase for action potential firing. Thus Zn(2+) may modulate neuronal excitability normally and in pathological conditions such as seizures by potentiating GlyRs tonically activated by low agonist concentrations. PMID- 17913993 TI - Modulation of soleus H-reflexes during gait in children with cerebral palsy. AB - In healthy adults, soleus H-reflexes are rhythmically modulated and generally depressed during gait compared with rest. From ages 6 to 13 yr, there is a progressive increase in the tonic inhibition of H-reflexes during walking, especially during the stance phase of the step cycle. In adults, rhythmic modulation and tonic depression are severely disturbed after bilateral spinal lesions but remain partly preserved after unilateral cerebral lesions. Children with diplegic cerebral palsy (CP) suffer from a bilateral supraspinal lesion of the corticospinal tract that occurs before the maturation of the CNS is complete. If supraspinal structures are involved in the tonic, but not rhythmic, age dependent reflex depression, it could be hypothesized that the tonic reflex depression with age is disturbed in CP, whereas the rhythmic part of the modulation remains unaffected. To test this hypothesis, soleus H-reflexes were assessed during gait in 16 CP children aged 5-11 and 15-16 and compared with 25 age-matched healthy children walking at similar velocities. Although the rhythmic part of the modulation pattern was present in CP, there was no significant tonic reflex depression with age, thus reflecting a lack of maturation of the corticospinal tract. It is argued the rhythmic part of the modulation may be generated on a spinal or brain stem level and is therefore not affected by the bilateral supraspinal lesion, whereas the tonic depression that occurs with maturation of the CNS is under supraspinal control. In conclusion, the supraspinal structures affected in CP are therefore likely involved in this age dependent tonic depression. PMID- 17913994 TI - Hedonic-specific activity in piriform cortex during odor imagery mimics that during odor perception. AB - Although it is known that visual imagery is accompanied by activity in visual cortical areas, including primary visual cortex, whether olfactory imagery exists remains controversial. Here we asked whether cue-dependent olfactory imagery was similarly accompanied by activity in olfactory cortex, and in particular whether hedonic-specific patterns of activity evident in olfactory perception would also be present during olfactory imagery. We used functional magnetic resonance imaging to measure activity in subjects who alternated between smelling and imagining pleasant and unpleasant odors. Activity induced by imagining odors mimicked that induced by perceiving real odorants, not only in the particular brain regions activated, but also in its hedonic-specific pattern. For both real and imagined odors, unpleasant stimuli induced greater activity than pleasant stimuli in the left frontal portion of piriform cortex and left insula. These findings combine with findings from other modalities to suggest activation of primary sensory cortical structures during mental imagery of sensory events. PMID- 17913995 TI - Plasticity of the synaptic modification range. AB - Activity-dependent synaptic plasticity is likely to provide a mechanism for learning and memory. Cortical synaptic responses that are strengthened within a fixed synaptic modification range after 5 days of motor skill learning are driven near the top of their range, leaving only limited room for additional synaptic strengthening. If synaptic strengthening is a requisite step for acquiring new skills, near saturation of long-term potentiation (LTP) should impede further learning or the LTP mechanism should recover after single-task learning. Here we show that the initial learning-induced synaptic enhancement is sustained even long after training has been discontinued and that the synaptic modification range shifts upward. This range shift places increased baseline synaptic efficacy back within the middle of its operating range, allowing prelearning levels of LTP and long-term depression. Persistent synaptic strengthening might be a substrate for long-term retention in motor cortex, whereas the shift in synaptic modification range ensures the availability for new synaptic strengthening. PMID- 17913996 TI - Separate adaptive mechanisms for controlling trajectory and final position in reaching. AB - We examined control of the hand's trajectory (direction and shape) and final equilibrium position in horizontal planar arm movements by quantifying transfer of learned visuomotor rotations between two tasks that required aiming the hand to the same spatial targets. In a trajectory-reversal task ("slicing"), the hand reversed direction within the target and returned to the origin. In a positioning task ("reaching"), subjects moved the hand to the target and held it there; cursor feedback was provided only after movement ended to isolate learning of final position from trajectory direction. We asked whether learning acquired in one task would transfer to the other. Transfer would suggest that the hand's entire trajectory, including its endpoint, was controlled using a common spatial plan. Instead we found minimal transfer, suggesting that the brain used different representations of target position to specify the hand's initial trajectory and its final stabilized position. We also observed asymmetrical practice effects on hand trajectory, including systematic curvature of reaches made after rotation training and hypermetria of untrained slice reversals after reach training. These are difficult to explain with a unified control model, but were replicated in computer simulations that specified the hand's initial trajectory and its final equilibrium position. Our results suggest that the brain uses different mechanisms to plan the hand's initial trajectory and final position in point-to point movements, that it implements these control actions sequentially, and that trajectory planning does not account for specific impedance values to be implemented about the final stabilized posture. PMID- 17913997 TI - EBM and quality improvement research. PMID- 17913998 TI - Law & psychiatry: Death row delusions: when is a prisoner competent to be executed? AB - This column examines a recent U.S. Supreme Court decision in Panetti v. Quarterman, which embraced a broader view of what makes death row prisoners incompetent to be executed. Although the defendant understood that he was to be executed and the state's purported reason for seeking his death--two criteria suggested by the Court's 1986 decision in Ford v. Wainwright--he suffered from a fixed delusion about the actual reason for his death. The Court indicated that competent prisoners must have a "rational understanding" of the reason that a death penalty is being imposed but declined to define a clear standard. PMID- 17913999 TI - Datapoints: Use of information technology by psychiatrists and other medical providers. PMID- 17914000 TI - Alcohol & drug abuse: Revisiting employee assistance programs and substance use problems in the workplace: key issues and a research agenda. AB - This column describes employee assistance program (EAPs) and identifies key issues for contemporary EAPs. These programs began as occupational alcohol programs and have evolved into more comprehensive resources. To better understand contemporary EAPs, the authors suggest a research agenda that includes descriptive studies to provide an up-to-date picture of services; investigations of how contemporary EAPs address substance use problems, including management consultation for early identification; further study of EAPs' effects on outcomes, such as productivity and work group outcomes; examination of the relationship between EAPs and other workplace resources; further examination of influences on EAP utilization; and development and testing of EAP performance measures. PMID- 17914001 TI - Personal accounts: The system stole my identity. PMID- 17914002 TI - Cost-effectiveness of quality improvement programs for patients with subthreshold depression or depressive disorder. AB - OBJECTIVE: This study explored the cost-effectiveness of quality-improvement interventions for depression in primary care, relative to usual care, among patients with subthreshold depression or depressive disorder. METHODS: A total of 746 primary care patients in managed care organizations with 12-month depressive disorder and 502 with current depressive symptoms but no disorder (subthreshold depression) participated in a group-level randomized controlled trial initiated between June 1996 and March 1997. Matched clinics were randomly assigned to enhanced usual care or one of two quality improvement interventions that provided education to manage depression over time and resources to facilitate access to medication management or psychotherapy for six to 12 months. RESULTS: The cost effectiveness ratio for the pooled intervention groups versus usual care was $2,028 for patients with subthreshold depression (95% confidence interval [CI]= $17,225 to $21,282) and $53,716 for those with depressive disorder (CI=$14,194 to $93,238), by using a measure of quality-adjusted life years (QALY) based on the 12-Item Short Form Health Survey. Similar results were obtained when alternative QALY measures were used. CONCLUSIONS: Although precision was limited, even the upper limit of the 95% CIs suggests that such interventions are as cost-effective for patients with subthreshold depression as are many widely used medical therapies. Despite lack of evidence for efficacy of treatments for subthreshold depression, disease management programs that support clinical care decisions over time for patients with subthreshold depression or depressive disorder can yield cost-effectiveness ratios comparable to those of widely adopted medical therapies. Achieving greater certainty about average cost-effectiveness would require a much larger study. PMID- 17914003 TI - Fidelity outcomes in the National Implementing Evidence-Based Practices Project. AB - OBJECTIVE: This article presents fidelity outcomes for five evidence-based practices that were implemented in routine public mental health settings in the National Implementing Evidence-Based Practices Project. METHODS: Over a two-year period 53 community mental health centers across eight states implemented one of five evidence-based practices: supported employment, assertive community treatment, integrated dual disorders treatment, family psychoeducation, and illness management and recovery. An intervention model of practice dissemination guided the implementation. Each site used both human resources (consultant trainers) and material resource (toolkits) to aid practice implementation and to facilitate organizational changes. External assessors rated fidelity to the evidence-based practice model every six months from baseline to two years. RESULTS: More than half of the sites (29 of 53, or 55%) showed high-fidelity implementation at the end of two years. Significant differences in fidelity emerged by evidence-based practice. Supported employment and assertive community treatment had higher fidelity scores at baseline and across time. Illness management and recovery and integrated dual disorders treatment had lower scores on average throughout. In general, evidence-based practices showed an increase in fidelity from baseline to 12 months, with scores leveling off between 12 and 24 months. CONCLUSIONS: Most mental health centers implemented these evidence-based practices with moderate to high fidelity. The critical time period for implementation was approximately 12 months, after which few gains were made, although sites sustained their attained levels of evidence-based practice fidelity for another year. PMID- 17914004 TI - Estimating Medicare Part D's impact on medication access among dually eligible beneficiaries with mental disorders. AB - OBJECTIVE: Medicare drug benefit (Part D) plans may restrict psychotropic medications more than state Medicaid programs do. This may have important implications for patients dually eligible for Medicare and Medicaid whose medications previously were covered under Medicaid. The objective of this study was to estimate rates of medication switching among dually eligible beneficiaries using antidepressants, antipsychotics, and mood stabilizers attributable to their enrollment in Medicare drug plans. METHODS: Baseline data on medication usage patterns among 467 dually eligible beneficiaries with mental disorders from the Medicare Current Beneficiary Survey, formulary data from a sample of Medicare drug plans, and estimates of the utilization response to pharmacy management tools from the Medicaid literature were used to estimate the likelihood of switching medications conditional on use of drugs and assignment to particular Medicare drug plans. RESULTS: Restrictions on psychotropic medications were common among the drug plans studied. Estimated rates of medication switching attributable to Medicare Part D were 6%-10% among dually eligible beneficiaries using antipsychotics, 5%-7% among those using antidepressants, and 2%-4% among those using mood stabilizers. Switching rates varied substantially across plans. CONCLUSIONS: On average, relatively few dually eligible beneficiaries with mental disorders are likely to experience treatment disruptions because of formulary restrictions and utilization controls used by Medicare drug plans. However, beneficiaries in some plans will experience significant barriers to medication access. Given the substantial variation among Medicare drug plans' management of psychotropic medication use, clinicians and social service agencies should counsel their beneficiaries with mental disorders in navigating the complex Medicare drug plan market. PMID- 17914005 TI - Experience of VA psychiatrists with pharmaceutical detailing of antipsychotic medications. AB - OBJECTIVE: The interaction between physicians and the pharmaceutical industry has become a subject of increased interest and concern. This study surveyed a national sample of psychiatrists practicing within Department of Veterans Affairs (VA) medical centers in 2005. It specifically focused on the experiences of these physicians with representatives of the manufacturers of second-generation antipsychotics. METHODS: VA psychiatrists were invited by e-mail to complete a Web-based questionnaire about their contact with representatives of each of the relevant pharmaceutical companies. Respondents were then questioned about several potential assertions about treatment effectiveness, side effects, and costs of these drugs. RESULTS: Of the 1,833 potential participants, 639 (35%) visited the Web site and completed the questionnaire. Among the responders, 558 (87%) reported at least one contact with company representatives. In the year before the survey the percentage of respondents reporting contact with representatives of each individual company varied from 58% to 70%. The three most commonly reported assertions made at any time in the past through direct speech during those meetings were that the representative's second-generation antipsychotic resulted in "a decreased risk of extrapyramidal symptoms" (79%), "greater symptom reduction than placebo" (78%), or "better negative symptom control than conventional antipsychotics" (77%). Statements least likely to be reported included that drugs resulted in "better positive symptom control than conventional antipsychotics" (36%), "better positive or negative symptom control than another atypical antipsychotic" (38%), and "increased risk of the development of diabetes mellitus" (39%). CONCLUSIONS: Comparing assertions reportedly made to VA psychiatrists with package insert information suggests that many assertions made by drug company representatives are inconsistent with prescribing information approved by the U.S. Food and Drug Administration, although assertions consistent with package insert information were more common than inconsistent ones. PMID- 17914006 TI - The application of cognitive-behavioral therapy for psychosis in clinical and research settings. AB - OBJECTIVE: This study compared the practice of cognitive-behavioral therapy (CBT) for psychosis across research and routine clinical settings. METHODS: An observer rated adherence measure was used to compare the content of 40 therapy sessions of clients with positive psychotic symptoms. Twenty therapist-client dyads came from a research setting in the United Kingdom and 20 from three clinical settings, two in the United Kingdom and one in the United States. In the research setting CBT was provided by research clinical psychologists and trained local therapists. In the clinical settings CBT was part of a case management service by trained therapists. RESULTS: Therapist adherence to CBT for psychosis did not differ between the research and clinical settings. However, clinicians in the research settings scored significantly higher on items for schema work (z=-1.98, p<.05), relapse prevention interventions (z=-2.08, p<.05), and formulating a model of relapse (z=-2.61, p<.01). CONCLUSIONS: CBT for psychosis conducted in clinical settings was more strongly characterized by assessment of symptoms and work on coping strategies and less so by relapse prevention and schema-level work. Relapse prevention interventions and schema work could be considered more challenging for therapists to undertake. The findings suggest that therapists working in routine clinical settings are able to establish good therapeutic relationships with people with psychosis and to work on assessing and coping with their psychotic symptoms. However, some therapeutic approaches may be more challenging in this context. PMID- 17914007 TI - Risk of psychological difficulties among children raised by custodial grandparents. AB - OBJECTIVE: Although custodial grandchildren are believed to be at greater risk of emotional and behavioral problems than children in general, scant research has examined this important public health issue. METHODS: This study involves data from 733 custodial grandmothers participating in a study funded by the National Institute of Mental Health on custodial grandparenting and 9,878 caregivers from the 2001 National Health Interview Survey (NHIS) who completed the Strengths and Difficulties Questionnaire (SDQ) in reference to target children between ages four and 17. These two data sets were used to determine if custodial grandmothers report greater emotional and behavioral difficulties, as measured by the SDQ, for custodial grandchildren than do caregivers of children from the NHIS normative sample. RESULTS: Custodial grandchildren fared worse than children from the NHIS sample across all domains measured by the SDQ subscales, regardless of the child's gender and whether reporters were recruited by population-based or convenience sampling methods. Comparisons within the sample of 733 custodial grandmothers showed that higher levels of difficulties were reported when grandmothers were caring for boys, were recruited by convenience versus population-based sampling, and were white. Comparisons of the banded scores computed for each SDQ subscale suggested that custodial grandchildren have different cutoff points than children in the general population for a likely diagnosis of a psychiatric disorder. CONCLUSIONS: Similar to other children in kinship care arrangements, custodial grandchildren are reported by their caregivers to have higher levels of behavioral and emotional disturbances than children in the overall U.S. population. PMID- 17914008 TI - Posttraumatic stress disorder and psychiatric comorbidity among detained youths. AB - OBJECTIVE: This study examined the prevalence of posttraumatic stress disorder (PTSD) and comorbid psychiatric disorders among juvenile detainees. METHODS: The sample consisted of a stratified random sample of 898 youths aged ten to 18 years who were arrested and detained in Chicago. RESULTS: Among participants with PTSD, 93% had at least one comorbid psychiatric disorder; however, among those without PTSD, 64% had at least one comorbid psychiatric disorder. Over half (54%) of the participants with PTSD had two or more types of comorbid disorders--that is, affective, anxiety, behavioral, or substance use disorders--and 11% had all four types of comorbid disorders. Among males, having any psychiatric diagnosis significantly increased the odds of having comorbid PTSD. Among females, alcohol use disorder and both alcohol and drug use disorders significantly increased the odds of having PTSD. No significant difference in prevalence rates of PTSD was found between males and females with specific psychiatric disorders. The prevalence of any comorbid psychiatric disorder was significantly greater for males with PTSD than that for females with PTSD (OR=3.4, CI=1.1-10.6, p<.05). CONCLUSIONS: Detection of comorbid PTSD among detained youths must be improved. PTSD is often missed because traumatic experiences are rarely included in standard screens or volunteered by patients. When planning treatment, clinicians must consider ramifications of comorbid PTSD. PMID- 17914009 TI - Racial differences in antidepressant treatment preceding suicide in a Medicaid population. AB - OBJECTIVE: African Americans have less use of antidepressants than whites, suggesting racial differences in the diagnosis and treatment of mood disorders. This study assessed whether this racial disparity is present among patients with a very high risk of severe mood disorders--that is, those with completed suicides. METHODS: Completed suicides that occurred between 1986 and 2004 were identified by examining records of Tennessee Medicaid/TennCare enrollees aged 18 to 84 years who had no serious medical illness or chronic psychosis. Use of antidepressants in the 365 days before the suicide was ascertained from filled prescriptions. Unconditional logistic regression was used to adjust for racial differences in demographic characteristics and somatic comorbidity. RESULTS: The study included 127 African Americans and 1,379 whites who completed suicide. African Americans had significantly reduced odds of receiving treatments for mood disorders in the year preceding the suicide: 29% of African Americans had filled an antidepressant prescription, compared with 51% of whites (adjusted odds ratio=.43, 95% confidence interval=.26-.71, p<.001). In contrast, there was no significant difference between the two racial groups in the proportions filling prescriptions for antipsychotic medications (13% of African Americans and 11% of whites). These findings persisted after the analysis controlled for other comorbidities linked with suicide, including alcohol or substance abuse, seizure disorders, borderline personality disorder, and serious neurological conditions. CONCLUSIONS: Although there are several other potential explanations, the study findings provide indirect evidence that there is underdiagnosis or undertreatment of African Americans with serious mood disorders. Further research on this question in African-American populations is essential. PMID- 17914010 TI - The theory of planned behavior applied to continuing education for mental health professionals. AB - OBJECTIVE: This study evaluated the effects of a continuing education class that applied the theory of planned behavior to the intentions and behavior of mental health practitioners. METHODS: In 2006 a total of 94 mental health practitioners were randomly assigned to either a standard continuing education class or one that applied principles of the theory of planned behavior. The class topic was a brief, self-report tool that assesses felt need for employment among people with serious mental illnesses. Participants' intentions to apply the tool were evaluated before and after each class. Participants' implementation of the tool was measured three months after the class. RESULTS: The class guided by the theory of planned behavior significantly and substantially increased the participants' intentions to use the tool in comparison with the standard class. Significantly more participants in the theory-guided class than in the standard class (74% versus 42%) had applied the tool by the three-month follow-up. Among those who implemented the assessment tool, the participants in the theory of planned behavior class also assessed significantly more of their caseload. CONCLUSION: The theory of planned behavior can improve and may be well suited to continuing education in psychiatry. PMID- 17914011 TI - Transformation of children's mental health services: the role of school mental health. AB - The New Freedom Commission has called for a transformation in the delivery of mental health services in this country. The commission's report and recommendations have highlighted the role of school mental health services in transforming mental health care for children and adolescents. This article examines the intersection of school mental health programs and the commission's recommendations in order to highlight the role of school mental health in the transformation of the child and adolescent mental health system. Schools are uniquely positioned to play a central role in improving access to child mental health services and in supporting mental health and wellness as well as academic functioning of youths. The New Freedom Commission report articulated several goals related to school mental health: reducing stigma, preventing suicide, improving screening and treating co-occurring disorders, and expanding school mental health programs. The authors suggest strategies for change, including demonstrating relevance to schools, developing consensus among stakeholders, enhancing community mental health-school connections, building quality assessment and improvement, and considering the organizational context of schools. PMID- 17914012 TI - Schools' mental health responses after Hurricanes Katrina and Rita. AB - OBJECTIVE: After the displacement of students following Hurricanes Katrina and Rita, schools in several states enrolled many students with potential mental health needs. This study sought to understand how schools perceived the mental health needs of these students and what mental health programs they implemented. METHODS: Mental health personnel at 19 public schools or school systems and 11 private or parochial schools in Louisiana, Alabama, Texas, and Mississippi were interviewed at two time points (spring and fall-winter of 2006). RESULTS: Schools undertook diverse approaches to interventions, depending on the preexisting mental health infrastructure and personnel, the perceived needs of students, and the barriers or facilitators in each system. Interviewees described a rapid and comprehensive approach to the crisis in the immediate aftermath. Shortly afterward, some schools perceived little need for mental health services and refocused on their academic missions. Other school systems perceived student need but were unable to implement trauma-focused programs because staff were not prepared to deliver such services and funding was lacking. However, some systems and schools were able to implement new programs or extend programs to displaced students. Implementation challenges included difficulty communicating with parents, burnout among staff and program implementers, and efforts to balance the needs of the displaced students with those of the preexisting student population. CONCLUSIONS: Despite significant efforts to support students affected by the hurricanes, schools were limited in their ability to implement disaster-focused programs. Extension of crisis plans to include precrisis training in mental health programming for students and staff who have ongoing difficulties after a disaster or crisis may be beneficial. PMID- 17914013 TI - Stakeholders' perspectives on the recommendations of the President's New Freedom Commission on Mental Health. AB - OBJECTIVE: The purpose of this study was to understand stakeholder perspectives on school mental health and the mental health system as they relate to the goals identified by the President's New Freedom Commission on Mental Health. METHODS: A total of 11 focus groups were held in Maryland, Ohio, and New Mexico with groups of parents, youths, school- and community-based providers and staff, and child and school mental health advocates and leaders. Across the three sites, 105 individuals participated in the focus groups. RESULTS: The stakeholders provided several important recommendations to advance the field of mental health. They included addressing stigma, implementing culturally competent care, providing mental health training to school staff, and increasing collaboration between community providers. CONCLUSIONS: Obtaining the views of key stakeholders is critical to transforming the mental health system and expanding the focus on mental health in schools. PMID- 17914014 TI - Effect of patients' reasons for refusing treatment on implementing psychiatric advance directives. AB - OBJECTIVE: Clinicians have raised concerns that psychiatric advance directives may be used to refuse all treatment. However, people writing psychiatric advance directives can explicitly state their reasoning underlying treatment decisions. This study examined whether patients' reasons for refusing treatment influenced clinician decision making about implementing psychiatric advance directives. METHODS: A total of 597 mental health professionals completed a questionnaire that presented two scenarios: one in which the patient wrote a psychiatric advance directive refusing all medication because of paranoid delusions and one in which the patient wrote a psychiatric advance directive refusing all medication because of concerns about side effects. RESULTS: Twenty-two percent of clinicians reported that they would respect the former psychiatric advance directive, whereas 72% reported that they would respect the latter. After multivariate regression was used, the reason for treatment refusal remained the single significant predictor of clinicians' decision to honor a patient's psychiatric advance directive. CONCLUSIONS: Results show reasons for treatment refusal in psychiatric advance directives are likely to affect clinicians' decisions to implement the directives. PMID- 17914015 TI - Costs of implementing a computerized prescription system in a public mental health agency. AB - OBJECTIVE: The objective of this case report is to inform decision makers about costs associated with adding a computerized prescription component to an existing information system in specialty mental health agencies. METHODS: A computerized prescription system was implemented in four not-for-profit mental health agencies in an urban setting as part of a larger study looking at reducing racial disparities. This brief report describes the implementation costs at one agency with ten full-time-equivalent psychiatrists for which information was available on time devoted to implementation by the management information system personnel. The financial costs of the computer network hardware and software were also documented. RESULTS: The total initial cost was $27,607: preimplementation cost, $3,720; technology and system integration cost, $10,148; and training cost, $13,739. Annual ongoing cost was expected to be $14,725. CONCLUSIONS: The technology expenditure itself is not prohibitive for initial implementation as well as for ongoing support. PMID- 17914016 TI - Developing Iraq's mental health policy. AB - As Iraq faces the challenge of securing a sustainable resolution to the current violence, the burden of mental illness is likely to increase dramatically. The impact of Saddam Hussein's dictatorship, the Iran-Iraq war, U.S.-led economic sanctions, the Persian Gulf wars, and the U.S. invasion and subsequent violent insurgency have devastated Iraq's governmental and social infrastructure. Health care delivery across sectors has suffered greatly. During the reconstruction phase, the United States and coalition forces allocated resources to restructure Iraq's health care system. Many multinational organizations, governments, and policy makers had the political will as well as the financial and human resources to greatly influence Iraq's mental health program. However, the lack of an existing mental health plan stifled these efforts. Applying Kingdon's model for policy development, which includes political analysis, problem defining, and proposal drafting, the authors describe the development of Iraq's current mental health policy. PMID- 17914017 TI - An assessment of beliefs about mental health care among veterans who served in Iraq. AB - OBJECTIVE: This study assessed beliefs about mental health treatment in a group of soldiers newly returning from the war in Iraq. METHODS: Participants were 20 National Guard soldiers who had served in Operation Iraqi Freedom. Soldiers who in phone interviews screened positive for a mental disorder were asked about advantages and disadvantages of seeking treatment, who would or would not support treatment seeking, and facilitators and barriers to treatment seeking. RESULTS: Stigma was portrayed as a major disadvantage to treatment seeking. Yet most participants indicated that people would be supportive of treatment seeking. Reducing symptoms was a major advantage of care. Barriers, especially those viewed as "self-induced," such as pride, not being able to ask for help, and not being able to admit to having a problem, were considered major impediments. CONCLUSIONS: The findings suggest that interventions developed to engage veterans in care must be directed toward cognitive factors that motivate treatment seeking. PMID- 17914018 TI - Cultural competence reexamined: critique and directions for the future. AB - This Open Forum aims to stimulate productive dialogue about cultural competence in providing mental health care. The authors examine recent calls for culturally competent care in mental health practice and give a brief overview of the context in which demands for such care have arisen. Using select examples from anthropology, the authors provide evidence of the importance of culture in the production, presentation, and experience of psychic distress. Acknowledging the value of culturally appropriate care, the authors synthesize anthropological critiques of cultural competence models. The essay concludes with suggestions for future directions in cultural competence research and implementation. PMID- 17914019 TI - 2007 APA Gold Award: Integration of community psychiatry into primary care centers in Harris County, Texas. PMID- 17914020 TI - 2007 APA Gold Award: Helping youths with severe emotional disturbances at risk of institutionalization. PMID- 17914021 TI - 2007 APA Achievement Awards: Silver Award: Early Assessment and Support Team, Mid Valley Behavioral Care Network, Salem, Oregon--Early intervention for people with schizophrenia. PMID- 17914023 TI - Adding and switching antipsychotics. PMID- 17914024 TI - Seeking safety protocol for men and women. PMID- 17914025 TI - The role of Akt in the signaling pathway of the glycoprotein Ib-IX induced platelet activation. AB - The platelet von Willebrand factor (vWF) receptor, glycoprotein Ib-IX (GPIb-IX), mediates platelet adhesion and induces signaling leading to integrin activation. Phosphoinositol 3-kinase (PI3K) is important in GPIb-IX-mediated signaling. PI3K dependent signaling mechanisms, however, are unclear. We show that GPIb-IX induced platelet aggregation and stable adhesion under flow were impaired in mouse platelets deficient in PI3K effectors, Akt1 and Akt2, and in human platelets treated with an Akt inhibitor, SH-6. Akt1 and Akt2 play important roles in early GPIb-IX signaling independent of Syk, adenosine diphosphate (ADP), or thromboxane A2 (TXA2), in addition to their recognized roles in ADP- and TXA2 dependent secondary amplification pathways. Knockout of Akt1 or Akt2 diminished platelet spreading on vWF but not on immobilized fibrinogen. Thus, Akt1 and Akt2 are both required only in the GPIb-IX-mediated integrin activation (inside-out signaling). In contrast, PI3K inhibitors abolished platelet spreading on both vWF and fibrinogen, indicating a role for PI3K in integrin outside-in signaling distinct from that in GPIb-IX-mediated inside-out signaling. Furthermore, Akt1- or Akt2-deficiency diminished vWF-induced cGMP elevation, and their inhibitory effects on GPIb-IX-dependent platelet adhesion were reversed by exogenous cGMP. Thus, Akt1 and Akt2 mediate GPIb-IX signaling via the cGMP-dependent signaling pathway. PMID- 17914026 TI - Membrane cofactor protein mutations in atypical hemolytic uremic syndrome (aHUS), fatal Stx-HUS, C3 glomerulonephritis, and the HELLP syndrome. AB - The hemolytic uremic syndrome (HUS) is a triad of microangiopathic hemolytic anemia, thrombocytopenia, and renal impairment. Genetic studies demonstrate that heterozygous mutations of membrane cofactor protein (MCP;CD46) predispose to atypical HUS (aHUS), which is not associated with exposure to Shiga toxin (Stx). Among the initial 25 MCP mutations in patients with aHUS were 2, R69W and A304V, that were expressed normally and for which no dysfunction was found. The R69W mutation is in complement control protein module 2, while A304V is in the hydrophobic transmembrane domain. In addition to 3 patients with aHUS, the A304V mutation was identified in 1 patient each with fatal Stx-HUS, the HELLP (hemolysis, elevated liver enzymes, and low platelets) syndrome, and glomerulonephritis with C3 deposits. A major goal was to assess if these putative mutations lead to defective complement regulation. Permanent cell lines expressing the mutated proteins were complement "challenged," and membrane control of C3 fragment deposition was monitored. Both the R69W and A304V MCP mutations were deficient in their ability to control the alternative pathway of complement activation on a cell surface, illustrating the importance of modeling transmembrane proteins in situ. PMID- 17914027 TI - Signaling pathways governing stem-cell fate. AB - Hematopoietic stem cells (HSCs) are historically the most thoroughly characterized type of adult stem cell, and the hematopoietic system has served as a principal model structure of stem-cell biology for several decades. However, paradoxically, although HSCs can be defined by function and even purified to near homogeneity, the intricate molecular machinery and the signaling mechanisms regulating fate events, such as self-renewal and differentiation, have remained elusive. Recently, several developmentally conserved signaling pathways have emerged as important control devices of HSC fate, including Notch, Wingless-type (Wnt), Sonic hedgehog (Shh), and Smad pathways. HSCs reside in a complex environment in the bone marrow, providing a niche that optimally balances signals that control self-renewal and differentiation. These signaling circuits provide a valuable structure for our understanding of how HSC regulation occurs, concomitantly with providing information of how the bone marrow microenvironment couples and integrates extrinsic with intrinsic HSC fate determinants. It is the focus of this review to highlight some of the most recent developments concerning signaling pathways governing HSC fate. PMID- 17914029 TI - Relationship of polyunsaturated fatty acid intake to peripheral neuropathy among adults with diabetes in the National Health and Nutrition Examination Survey (NHANES) 1999 2004. AB - OBJECTIVE: This study investigated the association between dietary intake of polyunsaturated fatty acids (PUFAs) and peripheral neuropathy in the U.S. population. RESEARCH DESIGN AND METHODS: We analyzed data from the National Health and Nutrition Examination Survey (NHANES) 1999-2004 for adults >or=40 years of age with diagnosed diabetes, an assessment of peripheral neuropathy, and reliable 24-h dietary recall. The dietary intake of PUFAs was analyzed by peripheral neuropathy status. Multivariate logistic regression models were used to estimate the odds of having peripheral neuropathy in higher quintiles of PUFA intake compared with the lowest quintile. RESULTS: The mean dietary intake of linolenic acid was 1.25 +/- 0.07 g among adults with peripheral neuropathy, significantly lower than the 1.45 +/- 0.05 g intake among those without peripheral neuropathy. After controlling for potential confounding variables, adults whose linolenic acid intake was in the highest quintile had lower odds of peripheral neuropathy than adults in the lowest quintile (adjusted odds ratio 0.40 [95% CI 0.21-0.77]). CONCLUSIONS: Among adults with diagnosed diabetes, dietary intake of linolenic acid is positively associated with lower odds of peripheral neuropathy. PMID- 17914028 TI - Platelet-associated PF-4 as a biomarker of early tumor growth. AB - Early tumor detection and intervention are important determinants of survival in patients with cancer. We have recently reported that the "platelet angiogenesis proteome" may be used to detect microscopic tumors in mice. We now present evidence that changes in platelet-associated platelet factor-4 (PF-4) detect malignant growth across a spectrum of human cancers in mice. A deregulated expression of an 8206-Da protein was observed by surfaceenhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-ToF MS) proteomic comparison of platelets from normal and tumor-bearing mice. The differentially expressed protein was identified as PF-4 by tandem mass spectrometry and ProteinChip immunoassay using anti-PF-4 antibody. The platelet-associated PF-4 appeared to be up-regulated in early growth of human liposarcoma, mammary adenocarcinoma, and osteosarcoma. A 120-day follow-up study of liposarcoma revealed a sustained 2-fold or higher increase of platelet-associated PF-4 at 19, 30, and 120 days. In contrast, only an insignificant change of PF-4 was observed in the plasma of mice bearing the different human tumor xenografts, and throughout the 120 days of the liposarcoma study. We conclude that platelet associated PF-4, but not its plasma counterpart, may represent a potential biomarker of early tumor presence. PMID- 17914030 TI - Fatty acids and insulin modulate myocardial substrate metabolism in humans with type 1 diabetes. AB - OBJECTIVE: Normal human myocardium switches substrate metabolism preference, adapting to the prevailing plasma substrate levels and hormonal milieu, but in type 1 diabetes, the myocardium relies heavily on fatty acid metabolism for energy. Whether conditions that affect myocardial glucose use and fatty acid utilization, oxidation, and storage in nondiabetic subjects alter them in type 1 diabetes is not well known. RESEARCH DESIGN AND METHODS: To test the hypotheses that in humans with type 1 diabetes, myocardial glucose and fatty acid metabolism can be manipulated by altering plasma free fatty acid (FFA) and insulin levels, we quantified myocardial oxygen consumption (MVo(2)), glucose, and fatty acid metabolism in nondiabetic subjects and three groups of type 1 diabetic subjects (those studied during euglycemia, hyperlipidemia, and a hyperinsulinemic euglycemic clamp) using positron emission tomography. RESULTS: Type 1 diabetic subjects had higher MVo(2) and lower myocardial glucose utilization rate/insulin than control subjects. In type 1 diabetes, glucose utilization increased with increasing plasma insulin and decreasing FFA levels. Myocardial fatty acid utilization, oxidation, and esterification rates increased with increasing plasma FFA. Increasing plasma insulin levels decreased myocardial fatty acid esterification rates but increased the percentage of fatty acids going into esterification. CONCLUSIONS: Type 1 diabetes myocardium has increased MVo(2) and is insulin resistant during euglycemia. However, its myocardial glucose and fatty acid metabolism still responds to changes in plasma insulin and plasma FFA levels. Moreover, insulin and plasma FFA levels can regulate the intramyocardial fate of fatty acids in humans with type 1 diabetes. PMID- 17914032 TI - Comparison of the effects of pioglitazone and metformin on hepatic and extra hepatic insulin action in people with type 2 diabetes. AB - OBJECTIVE: To determine mechanisms by which pioglitazone and metformin effect hepatic and extra-hepatic insulin action. RESEARCH DESIGN AND METHODS: Thirty-one subjects with type 2 diabetes were randomly assigned to pioglitazone (45 mg) or metformin (2,000 mg) for 4 months. RESULTS: Glucose was clamped before and after therapy at approximately 5 mmol/l, insulin raised to approximately 180 pmol/l, C peptide suppressed with somatostatin, glucagon replaced at approximately 75 pg/ml, and glycerol maintained at approximately 200 mmol/l to ensure comparable and equal portal concentrations on all occasions. Insulin-induced stimulation of glucose disappearance did not differ before and after treatment with either pioglitazone (23 +/- 3 vs. 24 +/- 2 micromol x kg(-1) x min(-1)) or metformin (22 +/- 2 vs. 24 +/- 3 micromol x kg(-1) x min(-1)). In contrast, pioglitazone enhanced (P < 0.01) insulin-induced suppression of both glucose production (6.0 +/- 1.0 vs. 0.2 +/- 1.6 micromol x kg(-1) x min(-1)) and gluconeogenesis (n = 11; 4.5 +/- 0.9 vs. 0.8 +/- 1.2 micromol x kg(-1) x min(-1)). Metformin did not alter either suppression of glucose production (5.8 +/- 1.0 vs. 5.0 +/- 0.8 micromol x kg(-1) x min(-1)) or gluconeogenesis (n = 9; 3.7 +/- 0.8 vs. 2.6 +/- 0.7 micromol x kg(-1) x min(-1)). Insulin-induced suppression of free fatty acids was greater (P < 0.05) after treatment with pioglitazone (0.14 +/- 0.03 vs. 0.06 +/- 0.01 mmol/l) but unchanged with metformin (0.12 +/- 0.03 vs. 0.15 +/- 0.07 mmol/l). CONCLUSIONS: Thus, relative to metformin, pioglitazone improves hepatic insulin action in people with type 2 diabetes, partly by enhancing insulin-induced suppression of gluconeogenesis. On the other hand, both drugs have comparable effects on insulin-induced stimulation of glucose uptake. PMID- 17914033 TI - Denervation and high-fat diet reduce insulin signaling in T-tubules in skeletal muscle of living mice. AB - OBJECTIVE: Insulin stimulates muscle glucose transport by translocation of GLUT4 to sarcolemma and T-tubules. Despite muscle glucose uptake playing a major role in insulin resistance and type 2 diabetes, the temporal and spatial changes in insulin signaling and GLUT4 translocation during these conditions are not well described. RESEARCH DESIGN AND METHODS: We used time-lapse confocal imaging of green fluorescent protein (GFP) ADP-ribosylation factor nucleotide-binding site opener (ARNO) (evaluation of phosphatidylinositide 3-kinase activation) and GLUT4 GFP-transfected quadriceps muscle in living, anesthetized mice either muscle denervated or high-fat fed. T-tubules were visualized with sulforhodamine B dye. In incubated muscle, glucose transport was measured by 2-deoxy-D-[(3)H]-glucose uptake, and functional detubulation was carried out by osmotic shock. Muscle fibers were immunostained for insulin receptors. RESULTS: Denervation and high fat diet reduced insulin-mediated glucose transport. In denervated muscle, insulin-stimulated phosphatidylinositol 3,4,5 P(3) (PIP3) production was abolished in T-tubules, while PIP3 production at sarcolemma was increased 2.6 fold. Correspondingly, GLUT4-GFP translocation to T-tubules was abolished, while translocation to sarcolemma was increased 2.3-fold. In high fat-fed mice, a approximately 65% reduction in both insulin-induced T-tubular PIP3 production and GLUT4-GFP translocation was seen. Sarcolemma was less affected, with reductions of approximately 40% in PIP3 production and approximately 15% in GLUT4-GFP translocation. Access to T-tubules was not compromised, and insulin receptor distribution in sarcolemma and T-tubules was unaffected by denervation or high fat feeding. Detubulation of normal muscle reduced basal and abolished insulin induced glucose transport. CONCLUSIONS: Our findings demonstrate, for the first time, that impaired insulin signaling and GLUT4 translocation is compartmentalized in muscle and primarily localized to T-tubules and not sarcolemma during insulin resistance. PMID- 17914031 TI - Multiple superoxide dismutase 1/splicing factor serine alanine 15 variants are associated with the development and progression of diabetic nephropathy: the Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications Genetics study. AB - BACKGROUND: Despite familial clustering of nephropathy and retinopathy severity in type 1 diabetes, few gene variants have been consistently associated with these outcomes. RESEARCH DESIGN AND METHODS: We performed an individual-based genetic association study with time to renal and retinal outcomes in 1,362 white probands with type 1 diabetes from the Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications (DCCT/EDIC) study. Specifically, we genotyped 1,411 SNPs that capture common variations in 212 candidate genes for long-term complications and analyzed them for association with the time from DCCT baseline to event for renal and retinal outcomes using multivariate Cox proportion hazards models. To address multiple testing and assist interpretation of the results, false discovery rate q values were calculated separately for each outcome. RESULTS: We observed association between rs17880135 in the 3' region of superoxide dismutase 1 (SOD1) and the incidence of both severe nephropathy (hazard ratio [HR] 2.62 [95% CI 1.64-4.18], P = 5.6 x 10( 5), q = 0.06) and persistent microalbuminuria (1.82 [1.29-2.57], P = 6.4 x 10( 4), q = 0.46). Sequencing and fine-mapping identified additional SOD1 variants, including rs202446, rs9974610, and rs204732, which were also associated (P < 10( 3)) with persistent microalbuminuria, whereas rs17880135 and rs17881180 were similarly associated with the development of severe nephropathy. Attempts to replicate the findings in three cross-sectional case-control studies produced equivocal results. We observed no striking differences between risk genotypes in serum SOD activity, serum SOD1 mass, or SOD1 mRNA expression in lymphoblastoid cell lines. CONCLUSIONS: Multiple variations in SOD1 are significantly associated with persistent microalbuminuria and severe nephropathy in the DCCT/EDIC study. PMID- 17914035 TI - Open clinical trials. PMID- 17914034 TI - Defective lipid delivery modulates glucose tolerance and metabolic response to diet in apolipoprotein E-deficient mice. AB - OBJECTIVE: Apolipoprotein E (ApoE) regulates plasma lipid levels via modulation of lipolysis and serving as ligand for receptor-mediated clearance of triglyceride (TG)-rich lipoproteins. This study tested the impact of modulating lipid delivery to tissues on insulin responsiveness and diet-induced obesity. RESEARCH DESIGN AND METHODS: ApoE(+/+) and apoE(-/-) mice were placed on high-fat high-sucrose diabetogenic diet or control diet for 24 weeks. Plasma TG clearance, glucose tolerance, and tissue uptake of dietary fat and glucose were assessed. RESULTS: Plasma TG clearance and lipid uptake by adipose tissue were impaired, whereas glucose tolerance was improved in control diet-fed apoE(-/-) mice compared with apoE(+/+) mice after an oral lipid load. Fat mass was reduced in apoE(-/-) mice compared with apoE(+/+) mice under both dietary conditions. The apoE(-/-) mice exhibited lower body weight and insulin levels than apoE(+/+) mice when fed the diabetogenic diet. Glucose tolerance and uptake by muscle and brown adipose tissue (BAT) was also improved in mice lacking apoE when fed the diabetogenic diet. Indirect calorimetry studies detected no difference in energy expenditure and respiratory quotient between apoE(+/+) and apoE(-/-) mice on control diet. Energy expenditure and uncoupling protein-1 expression in BAT were slightly but not significantly increased in apoE(-/-) mice on diabetogenic diet. CONCLUSIONS: These results demonstrated that decreased lipid delivery to insulin sensitive tissues improves insulin sensitivity and ameliorates diet-induced obesity. PMID- 17914036 TI - Commercializing clinical trials--risks and benefits of the CRO boom. PMID- 17914037 TI - The development of prosthetic heart valves--lessons in form and function. PMID- 17914038 TI - Effectiveness of influenza vaccine in the community-dwelling elderly. AB - BACKGROUND: Reliable estimates of the effectiveness of influenza vaccine among persons 65 years of age and older are important for informed vaccination policies and programs. Short-term studies may provide misleading pictures of long-term benefits, and residual confounding may have biased past results. This study examined the effectiveness of influenza vaccine in seniors over the long term while addressing potential bias and residual confounding in the results. METHODS: Data were pooled from 18 cohorts of community-dwelling elderly members of one U.S. health maintenance organization (HMO) for 1990-1991 through 1999-2000 and of two other HMOs for 1996-1997 through 1999-2000. Logistic regression was used to estimate the effectiveness of the vaccine for the prevention of hospitalization for pneumonia or influenza and death after adjustment for important covariates. Additional analyses explored for evidence of bias and the potential effect of residual confounding. RESULTS: There were 713,872 person-seasons of observation. Most high-risk medical conditions that were measured were more prevalent among vaccinated than among unvaccinated persons. Vaccination was associated with a 27% reduction in the risk of hospitalization for pneumonia or influenza (adjusted odds ratio, 0.73; 95% confidence interval [CI], 0.68 to 0.77) and a 48% reduction in the risk of death (adjusted odds ratio, 0.52; 95% CI, 0.50 to 0.55). Estimates were generally stable across age and risk subgroups. In the sensitivity analyses, we modeled the effect of a hypothetical unmeasured confounder that would have caused overestimation of vaccine effectiveness in the main analysis; vaccination was still associated with statistically significant--though lower--reductions in the risks of both hospitalization and death. CONCLUSIONS: During 10 seasons, influenza vaccination was associated with significant reductions in the risk of hospitalization for pneumonia or influenza and in the risk of death among community-dwelling elderly persons. Vaccine delivery to this high-priority group should be improved. PMID- 17914039 TI - Donepezil for the treatment of agitation in Alzheimer's disease. AB - BACKGROUND: Agitation is a common and distressing symptom in patients with Alzheimer's disease. Cholinesterase inhibitors improve cognitive outcomes in such patients, but the benefits of these drugs for behavioral disturbances are unclear. METHODS: We randomly assigned 272 patients with Alzheimer's disease who had clinically significant agitation and no response to a brief psychosocial treatment program to receive 10 mg of donepezil per day (128 patients) or placebo (131 patients) for 12 weeks. The primary outcome was a change in the score on the Cohen-Mansfield Agitation Inventory (CMAI) (on a scale of 29 to 203, with higher scores indicating more agitation) at 12 weeks. RESULTS: There was no significant difference between the effects of donepezil and those of placebo on the basis of the change in CMAI scores from baseline to 12 weeks (estimated mean difference in change [the value for donepezil minus that for placebo], -0.06; 95% confidence interval [CI], -4.35 to 4.22). Twenty-two of 108 patients (20.4%) in the placebo group and 22 of 113 (19.5%) in the donepezil group had a reduction of 30% or greater in the CMAI score (the value for donepezil minus that for placebo, -0.9 percentage point; 95% CI, -11.4 to 9.6). There were also no significant differences between the placebo and donepezil groups in scores for the Neuropsychiatric Inventory, the Neuropsychiatric Inventory Caregiver Distress Scale, or the Clinician's Global Impression of Change. CONCLUSIONS: In this 12 week trial, donepezil was not more effective than placebo in treating agitation in patients with Alzheimer's disease. (ClinicalTrials.gov number, NCT00142324 [ClinicalTrials.gov].). PMID- 17914040 TI - Effectiveness and safety of drug-eluting stents in Ontario. AB - BACKGROUND: The placement of drug-eluting stents decreases the frequency of repeat revascularization procedures in patients undergoing percutaneous coronary intervention (PCI) in randomized clinical trials. However, there is uncertainty about the effectiveness of drug-eluting stents, and increasing concern about their safety, in routine clinical practice. METHODS: From the Cardiac Care Network of Ontario's population-based clinical registry of all patients undergoing PCI in Ontario, Canada, we identified a well-balanced cohort of 3751 pairs of patients, matched on the basis of propensity score, who received either bare-metal stents alone or drug-eluting stents alone during an index PCI procedure between December 1, 2003, and March 31, 2005. The primary outcomes of the study were the rates of target-vessel revascularization, myocardial infarction, and death. RESULTS: The 2-year rate of target-vessel revascularization was significantly lower among patients who received drug eluting stents than among those who received bare-metal stents (7.4% vs. 10.7%, P<0.001). Drug-eluting stents were associated with significant reductions in the rate of target-vessel revascularization among patients with two or three risk factors for restenosis (i.e., presence of diabetes, small vessels [<3 mm in diameter], and long lesions [> or =20 mm]) but not among lower-risk patients. The 3-year mortality rate was significantly higher in the bare-metal-stent group than in the drug-eluting-stent group (7.8% vs. 5.5%, P<0.001), whereas the 2-year rate of myocardial infarction was similar in the two groups (5.2% and 5.7%, respectively; P=0.95). CONCLUSIONS: Drug-eluting stents are effective in reducing the need for target-vessel revascularization in patients at highest risk for restenosis, without a significantly increased rate of death or myocardial infarction. PMID- 17914041 TI - CT colonography versus colonoscopy for the detection of advanced neoplasia. AB - BACKGROUND: Advanced neoplasia represents the primary target for colorectal cancer screening and prevention. We compared the diagnostic yield from parallel computed tomographic colonography (CTC) and optical colonoscopy (OC) screening programs. METHODS: We compared primary CTC screening in 3120 consecutive adults (mean [+/-SD] age, 57.0+/-7.2 years) with primary OC screening in 3163 consecutive adults (mean age, 58.1+/-7.8 years). The main outcome measures included the detection of advanced neoplasia (advanced adenomas and carcinomas) and the total number of harvested polyps. Referral for polypectomy during OC was offered for all CTC-detected polyps of at least 6 mm in size. Patients with one or two small polyps (6 to 9 mm) also were offered the option of CTC surveillance. During primary OC, nearly all detected polyps were removed, regardless of size, according to established practice guidelines. RESULTS: During CTC and OC screening, 123 and 121 advanced neoplasms were found, including 14 and 4 invasive cancers, respectively. The referral rate for OC in the primary CTC screening group was 7.9% (246 of 3120 patients). Advanced neoplasia was confirmed in 100 of the 3120 patients in the CTC group (3.2%) and in 107 of the 3163 patients in the OC group (3.4%), not including 158 patients with 193 unresected CTC-detected polyps of 6 to 9 mm who were undergoing surveillance. The total numbers of polyps removed in the CTC and OC groups were 561 and 2434, respectively. There were seven colonic perforations in the OC group and none in the CTC group. CONCLUSIONS: Primary CTC and OC screening strategies resulted in similar detection rates for advanced neoplasia, although the numbers of polypectomies and complications were considerably smaller in the CTC group. These findings support the use of CTC as a primary screening test before therapeutic OC. PMID- 17914042 TI - Clinical practice. Osteoarthritis of the hip. PMID- 17914043 TI - A critical appraisal of "chronic Lyme disease". PMID- 17914044 TI - Images in clinical medicine. Medical mystery--an unusual complication of colonoscopy. PMID- 17914045 TI - Clinical problem-solving. Nothing to cough at--a 73-year-old man presented to the emergency department with a 4-day history of nonproductive cough that worsened at night. PMID- 17914046 TI - Influenza--the goal of control. PMID- 17914047 TI - Treatment of neuropsychiatric symptoms in patients with dementia. PMID- 17914048 TI - Inhaled nitric oxide in preterm infants--correction. PMID- 17914049 TI - Effect of aircraft-cabin altitude on passenger discomfort. PMID- 17914050 TI - Eosinophilic esophagitis. PMID- 17914051 TI - More on thunderstorms and iPods. PMID- 17914052 TI - Retro-orbital hemorrhage after thrombolysis for acute myocardial infarction. PMID- 17914053 TI - Treatment with convalescent plasma for influenza A (H5N1) infection. PMID- 17914054 TI - Tissue-engineered blood vessel for adult arterial revascularization. PMID- 17914055 TI - Identification of novel membrane-binding domains in multiple yeast Cdc42 effectors. AB - The Rho-type GTPase Cdc42 is a central regulator of eukaryotic cell polarity and signal transduction. In budding yeast, Cdc42 regulates polarity and mitogen activated protein (MAP) kinase signaling in part through the PAK-family kinase Ste20. Activation of Ste20 requires a Cdc42/Rac interactive binding (CRIB) domain, which mediates its recruitment to membrane-associated Cdc42. Here, we identify a separate domain in Ste20 that interacts directly with membrane phospholipids and is critical for its function. This short region, termed the basic-rich (BR) domain, can target green fluorescent protein to the plasma membrane in vivo and binds PIP(2)-containing liposomes in vitro. Mutation of basic or hydrophobic residues in the BR domain abolishes polarized localization of Ste20 and its function in both MAP kinase-dependent and independent pathways. Thus, Cdc42 binding is required but is insufficient; instead, direct membrane binding by Ste20 is also required. Nevertheless, phospholipid specificity is not essential in vivo, because the BR domain can be replaced with several heterologous lipid-binding domains of varying lipid preferences. We also identify functionally important BR domains in two other yeast Cdc42 effectors, Gic1 and Gic2, suggesting that cooperation between protein-protein and protein-membrane interactions is a prevalent mechanism during Cdc42-regulated signaling and perhaps for other dynamic localization events at the cell cortex. PMID- 17914056 TI - The golgin GCC88 is required for efficient retrograde transport of cargo from the early endosomes to the trans-Golgi network. AB - Retrograde transport pathways from early/recycling endosomes to the trans-Golgi network (TGN) are poorly defined. We have investigated the role of TGN golgins in retrograde trafficking. Of the four TGN golgins, p230/golgin-245, golgin-97, GCC185, and GCC88, we show that GCC88 defines a retrograde transport pathway from early endosomes to the TGN. Depletion of GCC88 in HeLa cells by interference RNA resulted in a block in plasma membrane-TGN recycling of two cargo proteins, TGN38 and a CD8 mannose-6-phosphate receptor cytoplasmic tail fusion protein. In GCC88 depleted cells, cargo recycling was blocked in the early endosome. Depletion of GCC88 dramatically altered the TGN localization of the t-SNARE syntaxin 6, a syntaxin required for endosome to TGN transport. Furthermore, the transport block in GCC88-depleted cells was rescued by syntaxin 6 overexpression. Internalized Shiga toxin was efficiently transported from endosomes to the Golgi of GCC88 depleted cells, indicating that Shiga toxin and TGN38 are internalized by distinct retrograde transport pathways. These findings have identified an essential role for GCC88 in the localization of TGN fusion machinery for transport from early endosomes to the TGN, and they have allowed the identification of a retrograde pathway which differentially selects TGN38 and mannose-6-phosphate receptor from Shiga toxin. PMID- 17914057 TI - Rab11 is required for membrane trafficking and actomyosin ring constriction in meiotic cytokinesis of Drosophila males. AB - Rab11 is a small GTPase that regulates several aspects of vesicular trafficking. Here, we show that Rab11 accumulates at the cleavage furrow of Drosophila spermatocytes and that it is essential for cytokinesis. Mutant spermatocytes form regular actomyosin rings, but these rings fail to constrict to completion, leading to cytokinesis failures. rab11 spermatocytes also exhibit an abnormal accumulation of Golgi-derived vesicles at the telophase equator, suggesting a defect in membrane-vesicle fusion. These cytokinesis phenotypes are identical to those elicited by mutations in giotto (gio) and four wheel drive (fwd) that encode a phosphatidylinositol transfer protein and a phosphatidylinositol 4 kinase, respectively. Double mutant analysis and immunostaining for Gio and Rab11 indicated that gio, fwd, and rab11 function in the same cytokinetic pathway, with Gio and Fwd acting upstream of Rab11. We propose that Gio and Fwd mediate Rab11 recruitment at the cleavage furrow and that Rab11 facilitates targeted membrane delivery to the advancing furrow. PMID- 17914058 TI - AP-1 and ARF1 control endosomal dynamics at sites of FcR mediated phagocytosis. AB - Phagocytosis, the mechanism of ingestion of large material and microorganisms, relies on actin polymerization and on the focal delivery of intracellular endocytic compartments. The molecular mechanisms involved in the formation and delivery of the endocytic vesicles that are recruited at sites of phagocytosis are not well characterized. Here we show that adaptor protein (AP)-1 but not AP-2 clathrin adaptor complexes are recruited early below the sites of particle attachment and are required for efficient receptor-mediated phagocytosis in murine macrophages. Clathrin, however, is not recruited with the AP complexes. We further show that the recruitment of AP-1-positive structures at sites of phagocytosis is regulated by the GTP-binding protein ARF1 but is not sensitive to brefeldin A. Furthermore, AP-1 depletion leads to increased surface levels of TNF alpha, a cargo known to traffic through the endosomes to the plasma membrane upon stimulation of the macrophages. Together, our results support a clathrin independent role for AP complexes in endosomal dynamics in macrophages by retaining some cargo proteins, a process important for membrane remodeling during phagocytosis. PMID- 17914059 TI - Synaptotagmin C2A loop 2 mediates Ca2+-dependent SNARE interactions essential for Ca2+-triggered vesicle exocytosis. AB - Synaptotagmins contain tandem C2 domains and function as Ca(2+) sensors for vesicle exocytosis but the mechanism for coupling Ca(2+) rises to membrane fusion remains undefined. Synaptotagmins bind SNAREs, essential components of the membrane fusion machinery, but the role of these interactions in Ca(2+)-triggered vesicle exocytosis has not been directly assessed. We identified sites on synaptotagmin-1 that mediate Ca(2+)-dependent SNAP25 binding by zero-length cross linking. Mutation of these sites in C2A and C2B eliminated Ca(2+)-dependent synaptotagmin-1 binding to SNAREs without affecting Ca(2+)-dependent membrane binding. The mutants failed to confer Ca(2+) regulation on SNARE-dependent liposome fusion and failed to restore Ca(2+)-triggered vesicle exocytosis in synaptotagmin-deficient PC12 cells. The results provide direct evidence that Ca(2+)-dependent SNARE binding by synaptotagmin is essential for Ca(2+)-triggered vesicle exocytosis and that Ca(2+)-dependent membrane binding by itself is insufficient to trigger fusion. A structure-based model of the SNARE-binding surface of C2A provided a new view of how Ca(2+)-dependent SNARE and membrane binding occur simultaneously. PMID- 17914060 TI - Caenorhabditis elegans prom-1 is required for meiotic prophase progression and homologous chromosome pairing. AB - A novel gene, prom-1, was isolated in a screen for Caenorhabditis elegans mutants with increased apoptosis in the germline. prom-1 encodes an F-box protein with limited homology to the putative human tumor suppressor FBXO47. Mutations in the prom-1 locus cause a strong reduction in bivalent formation, which results in increased embryonic lethality and a Him phenotype. Furthermore, retarded and asynchronous nuclear reorganization as well as reduced homologous synapsis occur during meiotic prophase. Accumulation of recombination protein RAD-51 in meiotic nuclei suggests disturbed repair of double-stranded DNA breaks. Nuclei in prom-1 mutant gonads timely complete mitotic proliferation and premeiotic replication, but they undergo prolonged delay upon meiotic entry. We, therefore, propose that prom-1 regulates the timely progression through meiotic prophase I and that in its absence the recognition of homologous chromosomes is strongly impaired. PMID- 17914061 TI - Updated research nosology for HIV-associated neurocognitive disorders. AB - In 1991, the AIDS Task Force of the American Academy of Neurology published nomenclature and research case definitions to guide the diagnosis of neurologic manifestations of HIV-1 infection. Now, 16 years later, the National Institute of Mental Health and the National Institute of Neurological Diseases and Stroke have charged a working group to critically review the adequacy and utility of these definitional criteria and to identify aspects that require updating. This report represents a majority view, and unanimity was not reached on all points. It reviews our collective experience with HIV-associated neurocognitive disorders (HAND), particularly since the advent of highly active antiretroviral treatment, and their definitional criteria; discusses the impact of comorbidities; and suggests inclusion of the term asymptomatic neurocognitive impairment to categorize individuals with subclinical impairment. An algorithm is proposed to assist in standardized diagnostic classification of HAND. PMID- 17914062 TI - Randomized controlled trial of an oral CGRP receptor antagonist, MK-0974, in acute treatment of migraine. AB - OBJECTIVE: To determine an effective and tolerable dose of a novel oral calcitonin gene-related peptide (CGRP) receptor antagonist, MK-0974, for the acute treatment of migraine. METHODS: Randomized, double-blind, parallel-group, clinical trial with a two-stage, adaptive, dose-ranging design. Patients were allocated to treat a moderate or severe migraine attack with MK-0974 (25, 50, 100, 200, 300, 400, or 600 mg), rizatriptan 10 mg, or placebo taken orally. The primary endpoint was pain relief (reduction to mild or none) 2 hours after dosing. Secondary endpoints included pain freedom at 2 hours and sustained pain relief at 24 hours. A prespecified, blinded, automated interim analysis was used to discontinue randomization to less effective doses. RESULTS: Per the adaptive study design, the four lowest MK-0974 groups (25, 50, 100, 200 mg) were discontinued due to insufficient efficacy. For the remaining treatment groups, the estimated pain relief proportions at 2 hours were 300 mg (n = 38) 68.1%, 400 mg (n = 45) 48.2%, 600 mg (n = 40) 67.5%, rizatriptan 10 mg (n = 34) 69.5%, and placebo (n = 115) 46.3%. The prespecified primary efficacy hypothesis test, which compared the average 2-hour pain relief response proportion of the combined 300, 400, and 600 mg MK-0974 groups to placebo, was significant (P = 0.015). A generally similar efficacy pattern was seen for other endpoints. MK-0974 was generally well tolerated and there did not appear to be an increase in adverse events with increasing dose. CONCLUSIONS: The novel, orally administered calcitonin gene-related peptide (CGRP) receptor antagonist, MK-0974, was effective and generally well tolerated for the acute treatment of migraine. PMID- 17914063 TI - Cortical demyelination in PML and MS: Similarities and differences. AB - OBJECTIVE: To characterize pathologic changes in the cerebral cortex of patients with multiple sclerosis (MS) and progressive multifocal leukoencephalopathy (PML). METHODS: Autopsy brain tissue was obtained from 13 patients with PML, 4 patients with MS, 2 patients with HIV encephalopathy, and 1 subject without neurologic pathology. Immunohistochemistry for myelin proteins, inflammatory cells, and neurofilaments was performed to evaluate the distribution of cortical lesions, their inflammatory activity, and neuritic pathology. Confocal microscopy was applied to examine pathologic changes in neurites in PML cortex. RESULTS: Leukocortical, intracortical, and subpial patterns of cortical demyelination were represented in MS brain tissue. In PML brain tissue intracortical and leukocortical but not subpial lesions were observed. Cortical lesions in PML and MS contained fewer inflammatory cells than demyelinated areas in the white matter. Neuritic pathology in cortical PML lesions was represented by dystrophic and transected neurites. Pathologic modifications in neuritic processes in PML were more evident in highly inflamed white matter than in gray matter areas of demyelination, reminiscent of previous reports of neuritic pathology in MS. JC virus-infected cells were associated with PML white matter, leukocortical and intracortical lesions. CONCLUSIONS: Cortical pathology represents a distinct feature of progressive multifocal leukoencephalopathy. Similarities and differences with regard to multiple sclerosis cortical pathology were noted and may be informative regarding the pathogenesis of both disorders. PMID- 17914064 TI - Corticobasal syndrome and primary progressive aphasia as manifestations of LRRK2 gene mutations. AB - BACKGROUND: Mutations in the LRRK2 gene are an important cause of familial and nonfamilial parkinsonism. Despite pleomorphic pathology, LRRK2 mutations are believed to manifest clinically as typical Parkinson disease (PD). However, most genetic screens have been limited to PD clinic populations. OBJECTIVE: To clinically characterize LRRK2 mutations in cases recruited from a spectrum of neurodegenerative diseases. METHODS: We screened for the common G2019S mutation and several additional previously reported LRRK2 mutations in 434 individuals. A total of 254 patients recruited from neurodegenerative disease clinics and 180 neurodegenerative disease autopsy cases from the University of Pennsylvania brain bank were evaluated. RESULTS: Eight cases were found to harbor a LRRK2 mutation. Among patients with a mutation, two presented with cognitive deficits leading to clinical diagnoses of corticobasal syndrome and primary progressive aphasia. CONCLUSION: The clinical presentation of LRRK2-associated neurodegenerative disease may be more heterogeneous than previously assumed. PMID- 17914065 TI - Plasma amyloid beta protein is elevated in late-onset Alzheimer disease families. AB - OBJECTIVE: Plasma A beta levels are elevated in early-onset Alzheimer disease (AD) caused by autosomal dominant mutations. Our objective was to determine whether similar genetic elevations exist in late-onset AD (LOAD). METHODS: We measured plasma A beta in first-degree relatives of patients with LOAD in a cross sectional series and in extended LOAD families. We screened these subjects for pathogenic mutations in early-onset AD genes and determined their ApoE genotypes. RESULTS: Plasma A beta is significantly elevated in the LOAD first-degree relatives in comparison to unrelated controls and married-in spouses. These elevations are not due to ApoE epsilon 4 or pathogenic coding mutations in the known early-onset AD genes. CONCLUSIONS: The findings provide strong evidence for the existence of novel, as yet unknown genetic factors that affect late-onset Alzheimer disease by increasing A beta. PMID- 17914066 TI - Longitudinally preserved psychomotor performance in long-term asymptomatic HIV infected individuals. AB - BACKGROUND: Recent case reports have suggested that some asymptomatic HIV infected individuals can develop CNS disturbances despite intact immunologic functioning and long-term suppression of plasma HIV concentrations to undetectable levels. This possibility has not yet been systematically studied longitudinally. METHODS: Using longitudinal data from the Multicenter AIDS Cohort Study, we investigated neuropsychological performance in long-term asymptomatic HIV-infected men who have sex with men. Performance over a 5-year period on the Symbol Digit Modalities test and the Trail Making Tests were compared in three HIV-positive asymptomatic groups [defined as 1) highly active antiretroviral therapy (HAART) treated with undetectable viral loads (n = 83), 2) AIDS-free for more than 15 years without HAART (n = 29), and 3) absence of clinical AIDS or CD4(+) lymphocyte count below 200 cells/muL at the beginning and end of the study period (n = 233)] and in HIV-negative controls (n = 237). Data were analyzed using linear mixed models and proportional odds logistic regression modeling with generalized estimating equations. RESULTS: There was no evidence of performance differences or performance declines over the 5-year period of study in any of the three long-term asymptomatic groups as compared with the HIV-negative group in the Symbol Digit Modalities test or the Trail Making Tests. Performance decrements were, however, observed with increasing age in each of the tests administered, demonstrating that performance declines could be detected by these methods. CONCLUSIONS: Regardless of how long-term asymptomatic status was defined immunologically or virologically, neuropsychological test performances remained stable. These findings suggest that psychomotor speed is preserved over many years in HIV-infected individuals with controlled HIV viremia. PMID- 17914067 TI - Clinical trial outcome in neuropathic pain: relationship to study characteristics. AB - BACKGROUND: Several recent randomized clinical trials have found that the medications being evaluated for neuropathic pain did not significantly differ from placebo for the primary efficacy endpoint, despite encouraging results from prior preclinical and clinical studies. It is unclear whether these trials were unsuccessful because the medications truly lack efficacy or whether characteristics of the trials compromised the demonstration of treatment benefits. OBJECTIVE: To identify factors associated with positive (i.e., favors medication) vs negative outcomes of placebo-controlled neuropathic pain trials. METHODS: We examined study characteristics associated with positive vs negative clinical trial outcomes for neuropathic pain treatments using the information provided in a comprehensive meta-analysis and additional ratings for 106 clinical trials. RESULTS: Univariate analyses indicated that the results of medication vs placebo comparisons were more likely to be positive when medication response rates were greater, placebo response rates were lower, and studies were published earlier. In a multivariate analysis performed to identify independent contributions of study characteristics to trial outcomes, greater medication response, reduced placebo response, and larger sample sizes were each uniquely associated with positive outcomes. In addition, greater medication response rates and parallel groups designs were each independently associated with greater placebo response rates. CONCLUSIONS: The results suggest that study characteristics may contribute to the outcomes of clinical trials of treatments for neuropathic pain and provide an impetus for investigating strategies for decreasing placebo response rates and thereby possibly increasing the likelihood of positive outcomes in trials of efficacious treatments. PMID- 17914068 TI - Anaphylactic reactions to fibrin sealant injection for spontaneous spinal CSF leaks. PMID- 17914069 TI - Differential expression of cell cycle genes in rhesus monkey oocytes and embryos of different developmental potentials. AB - Correct cell cycle regulation is especially challenging at the start of life. Ovulated oocytes must maintain meiotic arrest until fertilization, and then complete meiosis and initiate a series of modified cell divisions without growth. Moreover, myriad key developmental events, such as chromatin remodeling and transcriptional activation of the genome, are coordinated with each other via the cell cycle, particularly passage through the DNA synthesis phase (S Phase). We examined here the expression of more than 30 mRNAs related to cell cycle regulation in rhesus monkey oocytes and embryos and compared the expression of these mRNAs between oocytes and embryos of different developmental potentials. We find that the maternally inherited stores of cell cycle regulatory mRNAs are especially susceptible to disruption in cases of diminished oocyte and embryo quality in the rhesus monkey. In comparison to published mouse array data, we also observed striking species differences in the temporal expression patterns of many of these genes, suggesting that mechanisms of cell cycle control may differ and that the responses of oocytes and embryos to external insults may likewise differ. PMID- 17914070 TI - Regulation of Pcsk6 expression during the preantral to antral follicle transition in mice: opposing roles of FSH and oocytes. AB - Several secreted products of the TGFbeta superfamily have important roles during follicular development and are produced by both oocytes and somatic cells (granulosa and theca) in the follicle. The proprotein convertases are a family of seven known proteins that process TGFbeta ligands and other secreted products to their mature active form. The present study examined the regulation of steady state levels of Pcsk6 mRNA, which encodes a convertase protein known to process members of the TGFbeta superfamily, during mouse follicular development. Pcsk6 mRNA and protein were expressed in preantral but not cumulus or mural granulosa cells. Pcsk6 mRNA levels in preantral granulosa cells were not regulated by growing oocytes of preantral follicles, but were elevated by FSH. Furthermore, Pcsk6 mRNA in preantral granulosa cells was potently suppressed by factor(s) secreted by fully grown oocytes from antral follicles, in part through SMAD2/3 mediated pathways. Oocytes acquired the ability to suppress the steady-state levels of Pcsk6 mRNA in granulosa cells during the preantral to antral follicle transition. Suppression of Pcsk6 mRNA by oocytes could reflect a change in the mechanism(s) regulating the activity of members of the TGFbeta superfamily. PMID- 17914071 TI - Betacellulin overexpression in the mouse ovary leads to MAPK3/MAPK1 hyperactivation and reduces litter size by impairing fertilization. AB - The epidermal growth factor receptor (EGFR) and its ligands are emerging as key molecules in regulating female reproduction. Here, we used a transgenic mouse model to evaluate whether and at which level of the reproduction cascade higher than-normal levels of the EGFR ligand betacellulin (BTC) in the reproductive organs affect fertility. Western blots and immunohistochemistry revealed increased BTC levels in uterus and ovaries from transgenic females, particularly evident in granulosa cells of antral follicles. Onset of puberty, estrous cyclicity, and the anatomy and histology of reproductive organs at puberty were not altered as compared to control females. Fertility tests revealed a reduction (~50%) in litter size as the major reproductive deficit of transgenic females. Embryo implantation was delayed in transgenic females, but this was not the reason for the reduced litter size. Transgenic females produced a normal number of oocytes after natural ovulation. The in vivo fertilization rate was significantly reduced in untreated transgenic females but returned to normal levels after superovulation. Impaired oocyte fertilization in the absence of superovulation treatment was associated with MAPK3/MAPK1 hyperactivation in BTC transgenic ovaries, whereas similar levels of MAPK3/MAPK1 activation were detected in transgenic and control ovaries after superovulation treatment. Thus, tight regulation of MAPK3/MAPK1 activity appears to be essential for appropriate granulosa cell function during oocyte maturation. Our study identified hitherto unknown effects of BTC overabundance in reproduction and suggests BTC as a novel candidate protein for the modulation of fertility. PMID- 17914072 TI - Kit ligand 2 promotes murine oocyte growth in vitro. AB - Oocyte-granulosa cell communication, mediated by paracrine factors, is essential for oocyte development. Kit ligand (KITL) is expressed in granulosa cells as soluble (KITL1) or membrane-associated (KITL2) proteins. However, the relative biopotency of each isoform during oocyte development is unknown. Our initial results showed that Kitl2 was down-regulated in cultured granulosa cells. To determine the effect of the two isoforms of KITL on oocyte growth, Kitl-deficient fibroblasts were transfected with constructs expressing either KITL1 or KITL2, and growing oocytes were isolated from 12-day-old mice and cultured on the transfected fibroblasts for 2 days. At the end of culture, oocyte diameters were measured, the incidence of spontaneous germinal vesicle breakdown (GVBD) was noted, and oocytes were analyzed for KIT receptor expression. Oocyte growth occurred only in the presence of the KITL2-producing fibroblasts, and suppression of KITL2 expression impaired oocyte growth. Up-regulation of KIT expression occurred in the presence of KITL2 but not KITL1. The presence of KITL2 inhibited spontaneous GVBD. Meiosis inhibitors did not attenuate the GVBD that occurred in the absence of KITL2, suggesting that this process reflects oocyte degeneration rather than meiotic progression. These results indicate that KITL2 is the principal KITL isoform required for oocyte growth and survival in vitro. PMID- 17914073 TI - Zoledronic acid and survival in breast cancer patients with bone metastases and elevated markers of osteoclast activity. AB - OBJECTIVE: Most breast cancer patients with bone metastases will receive bisphosphonate treatment. This post hoc analysis investigated whether early normalization of urinary N-telopeptide of type I collagen (NTX) levels during bisphosphonate therapy correlates with a long-term reduction in skeletal-related event (SRE) risk and mortality in patients with breast cancer. METHODS: This was a retrospective subset analysis of a phase III randomized trial comparing i.v. zoledronic acid with pamidronate treatment in patients with bone metastases from breast cancer or multiple myeloma. Patients with breast cancer who had NTX assessments at baseline and at months 1 and 3 (n = 342) were classified as normal (NTX < 64 nmol/mmol creatinine) or elevated (NTX > or = 64 nmol/mmol creatinine). The relative risk for an SRE or death was estimated with Cox regression models. RESULTS: Among the 328 evaluable patients treated with zoledronic acid, 196 patients (59.7%) had elevated baseline NTX, with 149 of those patients (76.0%) having normalized NTX levels and 31 patients (15.8%) having persistently elevated NTX levels at 3 months. The normalized NTX group had significantly lower risks for a first SRE, a first fracture or surgery to bone, or death than the group that had persistently elevated NTX levels. CONCLUSIONS: These results suggest that early normalization of elevated baseline NTX while receiving zoledronic acid is associated with longer event-free and overall survival times compared with persistently elevated NTX. Further analyses in cancer patients with elevated marker levels are warranted to confirm the implications of these findings. PMID- 17914074 TI - How to preserve fertility in young women exposed to chemotherapy? The role of GnRH agonist cotreatment in addition to cryopreservation of embrya, oocytes, or ovaries. AB - The possibilities to preserve fertility in women exposed to chemotherapy are: in vitro fertilization plus embryo cryopreservation, ovarian cryopreservation, unfertilized ova cryopreservation, and the administration of a gonadotropin releasing hormone (GnRH) agonist. Because none of these methods is ideal, combination of several methods should be considered. Because the chances of preserving gonadal function following combined-modality treatment are significantly better for girls than for boys, simulation of a prepubertal milieu was applied only to women of reproductive age. The administration of GnRH agonists to women with Hodgkin's disease, breast cancer, and other malignancies, or to patients with lupus nephropathy, in parallel with chemotherapy, by others and by us, has demonstrated a significantly lower rate of premature ovarian failure in survivors than in nonrandomized controls. Several prospective, randomized studies are ongoing. A recent meta-analysis found that the administration of a GnRH agonist, in addition to chemotherapy, to patients with breast cancer was associated with less recurrence and superior survival. Several possibilities to explain the beneficial effect of GnRH agonists to minimize chemotherapy-associated gonadotoxicity are suggested: (a) The hypogonadotropic milieu decreases the number of primordial follicles entering the differentiation stage, which is more vulnerable to chemotherapy; (b) The hypoestrogenic state decreases ovarian perfusion and delivery of chemotherapy to the ovaries; (c) A direct effect of the GnRH agonist on the ovary occurs independently of the gonadotropin level; (d) GnRH agonists may upregulate an intragonadal antiapoptotic molecule such as sphingosine-1-phosphate; (e) The GnRH agonist may protect ovarian germline stem cells. PMID- 17914075 TI - Absence of conclusive evidence for the safety and efficacy of gonadotropin releasing hormone analogue treatment in protecting against chemotherapy-induced gonadal injury. AB - Every year, an increasing number of women with malignant and nonmalignant diseases is successfully treated with cytotoxic chemotherapy and/or radiotherapy. Many of these patients suffer from infertility and gonadal failure as a result of these treatments. At present, these patients may resort to assisted-reproduction techniques to protect their future childbearing potential before the implementation of cytotoxic therapy. While embryo cryopreservation is an established technology, oocyte and ovarian tissue freezing techniques are still investigational. Nevertheless both of these techniques have resulted in live births. Apart from assisted-reproduction techniques, it has been extensively debated whether administration of gonadotropin-releasing hormone (GnRH) analogues in conjunction with chemotherapy can protect ovarian reserve against cytotoxic insult. In this manuscript, we debate the rationale for the effectiveness of GnRH analogue coadministration in preservation of fertility by reviewing the literature, and provide preliminary data to support our views. PMID- 17914076 TI - Commentary: Ovarian function does not equal fertility does not equal babies. PMID- 17914077 TI - Commentary: Oncologic drugs in patients with organ dysfunction: a summary. AB - There are few prospective data regarding the pharmacokinetics and clinical toxicity of commonly used chemotherapeutics in cancer patients with organ dysfunction. Although increasing numbers of studies are investigating newer chemotherapeutics in patients with liver or kidney dysfunction, most guidelines for dosing, especially for established agents, remain empiric. This review describes the available data (both prospective and case study) evaluating the impact of renal and hepatic dysfunction on toxicity and dosing of commonly used chemotherapeutics and provides a practical summary for their use in this setting. PMID- 17914078 TI - A mechanistic perspective of monoclonal antibodies in cancer therapy beyond target-related effects. AB - Several monoclonal antibodies are now in clinical use for cancer therapy, and many others are currently undergoing clinical evaluation. These agents offer unique specificity against key molecular targets on tumor cells or in the tumor microenvironment. The clinical efficacy of monoclonal antibodies is generally attributed to target-specific mechanisms resulting from neutralizing or inhibiting a growth factor or receptor that drives cell proliferation and tumor growth. Several targets, including CD20, human epidermal growth factor receptor 2, vascular endothelial growth factor, and epidermal growth factor receptor, have been validated in specific malignancies on the basis of monoclonal antibody efficacy. However, monoclonal antibodies also have the potential to activate immune-mediated effector functions, including antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity. These functions result from interactions involving the Fc domain of the antibody, and, consequently, may vary by antibody, isotype, and Fc modification, such as changes in glycosylation. Accordingly, all monoclonal antibodies directed against a given target should not be considered equivalent in their ability to stimulate immune-mediated effector functions. PMID- 17914079 TI - Small cell lung cancer: have we made any progress over the last 25 years? AB - Twenty-five years ago, small cell lung cancer was widely considered to be the next cancer added to the list of "curable cancers." This article attempts to summarize the progress made toward that goal since then. Clinical trials have provided landmarks in the therapy of limited-stage small cell lung cancer (LS SCLC). These are: (a) the proof that thoracic radiation therapy adds to systemic chemotherapy, (b) the superiority of twice-daily radiation therapy over daily fractionation, and (c) the need for prophylactic central nervous system radiation (prophylactic cranial irradiation). Each of these innovations adds about 5%-10% to the overall survival rate. In extensive-stage disease, irinotecan plus cisplatin may be a possible alternative to the "standard" etoposide-cisplatin chemotherapy doublet, but there has been little progress otherwise. It is imperative that, whenever possible, patients be given the opportunity to participate in future clinical trials so that the survival for these patients can continue to improve. PMID- 17914080 TI - Hope. AB - Shortly before his death in 1995, Kenneth B. Schwartz, a cancer patient at Massachusetts General Hospital (MGH), founded The Kenneth B. Schwartz Center at MGH. The Schwartz Center is a nonprofit organization dedicated to supporting and advancing compassionate health care delivery that provides hope to the patient, support to caregivers, and encourages the healing process. The Center sponsors the Schwartz Center Rounds, a monthly multidisciplinary forum where caregivers reflect on important psychosocial issues faced by patients, their families, and their caregivers, and gain insight and support from fellow staff members. A patient with recurrent ovarian cancer, now in a 12-year remission after recurrence, and her surgeon, discussed their experiences and feelings around the hopes and fears of cancer and its treatment. Hope sustains many through dark times, and is at the core of the wonderful resilience of many who wrestle with cancer. Concerns about false hope, unrealistic expectations, assumptions, engaging in realistic hopefulness, and the joys and stresses embodied in hope and how they frame the caregiver-patient relationship are discussed. The literature and limited evidence base are reviewed. PMID- 17914081 TI - Temozolomide for the treatment of metastatic melanoma: a systematic review. AB - BACKGROUND: This systematic review examines the role of temozolomide in patients with metastatic melanoma. Outcomes of interest include response rate, progression free survival, overall survival, quality of life, and adverse effects. METHODS: The MEDLINE, EMBASE, and Cochrane Library databases were searched from 1980 through to 2005 using variations on the search terms: melanoma, clinical trial, random, temozolomide, temodal, and temodar. The American Society of Clinical Oncology Annual Meeting proceedings were searched from 1996 to 2005. Relevant articles and abstracts were selected and reviewed by two reviewers, and the reference lists from these sources were searched for additional trials. RESULTS: Two randomized phase III trials and three randomized phase II trials were located. In addition, 21 phase I or II trials investigating single-agent temozolomide, temozolomide plus interferon-alpha, and temozolomide plus thalidomide were reviewed. A direct comparison of temozolomide and dacarbazine demonstrated equal efficacy for response rates and overall survival; however, no significant difference was reported. A second phase III study comparing single agent temozolomide with temozolomide combined with interferon-alpha indicated a significantly higher response rate for the combination treatment arm, but no difference in overall survival was noted. Further phase III studies are required to confirm whether there is a benefit associated with the combination of temozolomide and interferon-alpha or thalidomide. CONCLUSION: Our review of the available literature suggests that temozolomide demonstrates comparable activity to the current standard treatment, dacarbazine, with the additional benefit of being a convenient oral treatment that penetrates the blood-brain barrier. PMID- 17914082 TI - Early epirubicin-induced myocardial dysfunction revealed by serial tissue Doppler echocardiography: correlation with inflammatory and oxidative stress markers. AB - A phase II, open, nonrandomized trial was carried out in a group of epirubicin treated patients with cancer at different sites with the aim of detecting early preclinical changes that are predictive of the risk for heart failure. All patients underwent conventional echocardiography, as well as tissue Doppler imaging (TDI) with strain (sigma) and strain rate (SR), a very accurate technique for detecting minimal changes in cardiac left ventricular (LV) function. Moreover, echocardiographic changes identified during epirubicin treatment were compared with those of a series of biochemical markers of both myocardial damage and inflammation/oxidative stress. Sixteen patients (male-to-female ratio, 3:13; mean age +/- standard deviation, 56 +/-3 years; range, 27-75 years) with histologically confirmed tumors at different sites, scheduled to be treated with an epirubicin-based chemotherapy regimen, were enrolled in the study. A significant impairment in systolic LV function was observed after 200 mg/m2 of epirubicin; this was shown by a lower SR peak compared with baseline (1.82 +/- 0.57/second versus 1.45 +/- 0.44/second), whereas sigma remained unchanged. The following significant changes in LV diastolic function occurred only after 300 mg/m2 of epirubicin: a decrease in conventional early/late diastolic (E/A) velocities (1.16 +/- 0.31 versus 0.93 +/- 0.24) and a reduction in both the E(m) wave in the basal portion of the interventricular septum (8.86 +/- 1.73 cm/second versus 7.51 +/- 2.30 cm/second) and in the E(m)/A(m) ratio (1.09 +/- 0.51 versus 0.83 +/- 0.51), as measured using the TDI technique. No significant changes in LV ejection fraction were observed. Baseline values of brain natriuretic peptide, troponin I, myoglobin, and creatine kinase-myocardial subfraction were within the normal range and no significant changes were observed throughout the study. Levels of interleukin (IL)-6 and its soluble receptor (sIL-6R) and reactive oxygen species increased significantly, whereas glutathione peroxidase (GPx) levels decreased significantly, after 200 mg/m2 of epirubicin. Significant correlations between the reduction in the SR peak (deltaSR) after 200 mg/m2 of epirubicin and the increase in IL-6 and ROS and decrease in GPx were observed. The multiple regression analysis showed that the only independent predictive variable for deltaSR was ROS level. Our data show that: (a) subtle cardiac abnormalities may occur at epirubicin doses significantly below those known to be potentially clinically harmful and (b) the earliest myocardial impairment affects LV systolic rather than diastolic function. Early contractility impairment during epirubicin treatment was associated with high levels of ROS and markers of inflammation. The clinical meaningfulness of our findings warrants further investigations in a larger number of patients for a longer period of follow-up. PMID- 17914083 TI - Primary malignant sarcomas of the heart and great vessels in adult patients--a single-center experience. AB - BACKGROUND: Sarcomas arising in the heart or the great vessels are rare entities. The prognosis of the patients is dismal. METHODS: Between January 1993 and September 2006, of 1,429 patients registered to the Sarcoma Center, 14 had a primary sarcoma of the heart or large vessels. RESULTS: Tumors were located in the left ventricle (n = 3), left/right atrium (n = 2/3), pulmonary artery (n = 2), and ventricular septum, aorta, pericardium, and inferior vena cava (n = 1 each). The most frequently encountered histologic subtypes were leiomyosarcoma and angiosarcoma. Six patients presented with distant metastases to the lungs (n = 5), lymph nodes (n = 2), and liver (n = 1). Eight patients had localized disease. Six of them underwent resection with curative intent. Of those, two developed local recurrence within 2 and 10 months from surgery. Eleven patients received palliative chemotherapy, seven of those as initial treatment. Eight patients attained a response to treatment, two had disease stabilization for 6 and 12 months. After a median follow-up of 14.5 months (range, 2-156), three patients were alive with no evidence of disease 11, 52, and 156 months after diagnosis. Two patients were alive with disease and nine patients had died. CONCLUSIONS: Patients with primary sarcomas of the heart and the large vessels were of a young age, and more than half of them presented with advanced disease. Given the promising response to chemotherapy, an optimized treatment approach including neoadjuvant chemo-/radiotherapy in patients with locally advanced disease should be pursued. PMID- 17914084 TI - Guidelines for antiemetic treatment of chemotherapy-induced nausea and vomiting: past, present, and future recommendations. AB - Clinicians should be aware that chemotherapy-induced nausea and vomiting (CINV) is still one of the most feared side effects of chemotherapy. With the correct use of antiemetics, CINV can be prevented in almost 70% to up to 80% of patients. Treatment guidelines are useful tools that enable physicians to integrate the latest clinical research into their practices. The large volume of rapidly evolving clinical data has been summarized and incorporated into treatment recommendations by well-known and reliable institutions, including the Multinational Association of Supportive Care in Cancer, the American Society of Clinical Oncology, and the National Comprehensive Cancer Network. Despite the availability of such guidelines, however, there is evidence that adherence to and implementation of treatment recommendations are less than optimal. This review focuses, in particular, on the conformity and differences of these three guidelines. Furthermore, open questions and trends in the field of antiemesis are discussed as well. PMID- 17914085 TI - Randomized crossover pharmacokinetic evaluation of subcutaneous versus intravenous granisetron in cancer patients treated with platinum-based chemotherapy. AB - BACKGROUND: 5-HT3-receptor antagonists are one of the mainstays of antiemetic treatment, and they are administered either i.v. or orally. Nevertheless, sometimes neither administration route is feasible, such as in patients unable to admit oral intake managed in an outpatient setting. Our objective was to evaluate the bioavailability of s.c. granisetron. PATIENTS AND METHODS: Patients receiving platinum-based chemotherapy were randomized to receive 3 mg of granisetron either s.c. or i.v. in a crossover manner during two cycles. Blood and urine samples were collected after each cycle. Pharmacokinetic parameters observed with each administration route were compared by analysis of variance. RESULTS: From May to November 2005, 31 patients were included and 25 were evaluable. Subcutaneous granisetron resulted in a 27% higher area under the concentration-time curve for 0-12 hours (AUC(0-12h)) and higher levels at 12 hours, with similar values for AUC(0-24h). The maximum concentration was lower with the s.c. than with the i.v. route and was observed 30 minutes following s.c. administration. CONCLUSION: Granisetron administered s.c. achieves complete bioavailability. This is the first study that shows that s.c. granisetron might be a valid alternative to i.v. delivery. Further trials to confirm clinical equivalence are warranted. This new route of administration might be especially relevant for outpatient management of emesis in cancer patients. PMID- 17914086 TI - Late-onset epileptogenesis and seizure genesis: lessons from models of cerebral ischemia. AB - Patients surviving ischemic stroke often express delayed epileptic syndromes. Late poststroke seizures occur after a latency period lasting from several months to years after the insult. These seizures might result from ischemia-induced neuronal death and associated morphological and physiological changes that are only partly elucidated. This review summarizes the long-term morphofunctional alterations observed in animal models of both focal and global ischemia that could explain late-onset seizures and epileptogenesis. In particular, this review emphasizes the change in GABAergic and glutamatergic signaling leading to hyperexcitability and seizure genesis. PMID- 17914087 TI - Neuroendocrine-like prostate cancer cells: neuroendocrine transdifferentiation of prostate adenocarcinoma cells. AB - Neuroendocrine (NE) cells represent a minor cell population in the epithelial compartment of normal prostate glands and may play a role in regulating the growth and differentiation of normal prostate epithelia. In prostate tumor lesions, the population of NE-like cells, i.e., cells exhibiting NE phenotypes and expressing NE markers, is increased that correlates with tumor progression, poor prognosis, and the androgen-independent state. However, the origin of those NE-like cells in prostate cancer (PCa) lesions and the underlying molecular mechanism of enrichment remain an enigma. In this review, we focus on discussing the distinction between NE-like PCa and normal NE cells, the potential origin of NE-like PCa cells, and in vitro and in vivo studies related to the molecular mechanism of NE transdifferentiation of PCa cells. The data together suggest that PCa cells undergo a transdifferentiation process to become NE-like cells, which acquire the NE phenotype and express NE markers. Thus, we propose that those NE like cells in PCa lesions were originated from cancerous epithelial cells, but not from normal NE cells, and should be defined as 'NE-like PCa cells'. We further describe the biochemical properties of newly established, stable NE-like lymph node carcinoma of the prostate (LNCaP) cell lines, transdifferentiated from androgen-sensitive LNCaP cells under androgen-deprived conditions. Knowledge of understanding NE-like PCa cells will help us to explore new therapeutic strategies for treating PCa. PMID- 17914088 TI - Increased incidence of lobular breast cancer in women treated with hormone replacement therapy: implications for diagnosis, surgical and medical treatment. AB - A growing body of evidence support the association between the use of hormone replacement therapy (HRT) and a higher risk of both invasive lobular carcinoma (ILC) and invasive ductal-lobular mixed carcinoma (IDLC). Overall biological and clinical features of ILC entail a more cautious diagnostic and therapeutic approach as compared with invasive ductal carcinoma (IDC). ILCs are more frequently multifocal, multicentric and/or bilateral. Mammography and ultrasound show, therefore, significant limitations, while the higher sensitivity of magnetic resonance imaging in the detection of multifocal and/or multicentric lesions seems to improve the accuracy of preoperative staging of ILCs. Early diagnosis is even more challenging because the difficult in the localization and the sparse cellularity of lobular tumours may determine a false negative core biopsy. ILC is characterized by low proliferative activity, C-ErbB-2 negativity, bcl-2 positivity, p53 and VEGF negativity, oestrogen and progesterone positive receptors, low grade and low likelihood of lymphatic-vascular invasion. However, this more favourable biological behaviour does not reflect into a better disease free and overall survival as compared with IDC. Since lobular histology is associated with a higher risk of positive margins, mastectomy is often preferred to breast conservative surgery. Moreover, only few patients with ILC achieve a pathologic response to preoperative chemotherapy and, therefore, in most patients mastectomy can be regarded as the safer surgical treatment. The preoperative staging and the follow-up of patients with ILC are also complicated by the particular metastatic pattern of such histotype. In fact, metastases are more frequently distributed to the gastrointestinal tract, peritoneum/retroperitoneum and gynaecological organs than in IDC. PMID- 17914089 TI - The diagnosis and management of malignant phaeochromocytoma and paraganglioma. AB - Malignant phaeochromocytomas are rare tumours accounting for ~10% of all phaeochromocytomas; the prevalence of malignancy among paragangliomas is higher, especially those associated with succinate dehydrogenase subunit B gene mutations. Although a subset of these tumours has metastatic disease at initial presentation, a significant number develops metastases during follow-up after excision of an apparently benign tumour. Clinical, biochemical and histological features cannot reliably distinguish malignant from benign tumours. Although a number of recently introduced molecular markers have been explored, their clinical significance remains to be elucidated from further studies. Several imaging modalities have been utilised for the diagnosis and staging of these tumours. Functional imaging using radiolabelled metaiodobenzylguanidine (MIBG) and more recently, (18)F-fluorodopamine and (18)F-fluorodopa positron emission tomography offer substantial sensitivity and specificity to correctly detect metastatic phaeochromocytoma and paraganglioma and helps identify patients suitable for treatment with radiopharmaceuticals. The 5-year mortality rate of patients with malignant phaeochromocytomas and paragangliomas greater than 50% indicates that there is considerable room for the improvement of currently available therapies. The main therapeutic target is tumour reduction and control of symptoms of excessive catecholamine secretion. Currently, the best adjunctive therapy to surgery is treatment with radiopharmaceuticals using (131)I-MIBG; however, this is very rarely curative. Chemotherapy has been used for metastatic disease with only a partial and mainly palliative effect. The role of other forms of radionuclide treatment either alone or in combination with chemotherapy is currently evolving. Ongoing microarray studies may provide novel intracellular pathways of importance for proliferation/cell cycle control, and lead to the development of novel pharmacological agents. PMID- 17914090 TI - The optimal imaging of adrenal tumours: a comparison of different methods. AB - Computed tomography (CT; unenhanced, followed by contrast-enhanced examinations) is the cornerstone of imaging of adrenal tumours. Attenuation values of <10 Hounsfield units on an unenhanced CT are practically diagnostic for adenomas. When lesions cannot be characterised adequately with CT, magnetic resonance imaging (MRI) evaluation (with T1- and T2-weighted sequences and chemical shift and fat-suppression refinements) is sought. Functional nuclear medicine imaging is useful for adrenal lesions that are not adequately characterised with CT and MRI. Scintigraphy with [(131)I]-6-iodomethyl norcholesterol (a labelled cholesterol analogue) can differentiate adrenal cortical adenomas from carcinomas. Phaeochromocytomas appear as areas of abnormal and/or increased uptake of [(123)I]- and [(131)I]-meta-iodobenzylguanidine (a labelled noradrenaline analogue). The specific and useful roles of adrenal imaging include the characterisation of tumours, assessment of true tumour size, differentiation of adenomas from carcinomas and metastases, and differentiation of hyperfunctioning from non-functioning lesions. Adrenal imaging complements and assists the clinical and hormonal evaluation of adrenal tumours. PMID- 17914091 TI - Bicalutamide increases phospho-Akt levels through Her2 in patients with prostate cancer. AB - Bicalutamide monotherapy is emerging as an alternative in the treatment of locally advanced prostate cancer. However, a significant number of these patients will recur and be in need of second-line therapies. The knowledge of molecular arrangements after pharmacological therapy seems to be a new primary prerequisite to predict the efficacy or the failure of a secondary therapy. Based on these considerations, we have conducted this study in order to analyze the expressions of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), Akt, epidermal growth factor receptor (EGFR), phospho-EGFR (p-EGFR), human EGFR2 (Her2), and phospho-Her2 (p-Her2) after bicalutamide treatment. For this purpose, we evaluated retrospectively 69 prostate cancer tissues derived from patients who received radical prostatectomy as the only treatment, and 81 from patients who received bicalutamide for 120 days before surgery. In addition, we analyzed at different time points the effects of castration performed on athymic mice bearing the LuCaP 35 xenograft line at different times. We observed that bicalutamide treatment increased significantly the levels of p-Akt, EGFR, and Her2 with a concomitant reduction in PTEN. This effect was time dependent and required of sufficient time to be evident as indicated by data obtained with the LuCaP 35 tumors. A logistic multiple regression analysis revealed that a switch of p-Akt control from a PTEN/EGFR- to Her2-after bicalutamide treatment was possible. Since Akt and Her2 can be associated with reduced drug sensitivity, our report suggests that the evaluation of molecular arrangements after bicalutamide treatment could be useful to identify subsets of patients who will be molecular permissive for new adjuvant anti-target therapies. PMID- 17914092 TI - Expression of GnRH type II is regulated by the androgen receptor in prostate cancer. AB - GnRH II has important functional effects in steroid hormone-dependent tumours. Here we investigated the expression and regulation of GnRH II in prostate cancer. GnRH II protein was equally expressed in benign (73%) and malignant (78%) biopsies studied in a prostate tissue microarray (P = 0.779). There was no relationship between expression and clinical parameters in the cancer cohort. GnRH II was, however, significantly reduced in tumour biopsies following hormone ablation. This was further investigated in a prostate xenograft model where androgens increased GnRH II levels, while their withdrawal reduced it. In cell lines, we confirmed high levels of GnRH II in androgen receptor (AR)-positive LNCaP cells but low levels in AR-negative PC3 cells. In LNCaP cells, GnRH II induction by androgens was blocked by the AR inhibitor casodex, but not by cycloheximide treatment. Sequence analysis subsequently revealed a putative androgen response element in the upstream region of the GnRH II gene and direct interaction with the AR was confirmed in chromatin immunoprecipitation experiments. Finally, to test whether the effects of GnRH II were dependent on AR expression, LNCaP and PC3 cells were exposed to exogenous peptide. In both cell lines, GnRH II inhibited cell proliferation and migration, suggesting that its function is independent of AR status. These results provide evidence that GnRH II is widely expressed in prostate cancer and is an AR-regulated gene. Further studies are warranted to characterise the effects of GnRH II on prostate cancer cells and investigate its potential value as a novel therapy. PMID- 17914093 TI - Chromogranin A and biochemical progression-free survival in prostate adenocarcinomas submitted to radical prostatectomy. AB - The primary aim of the present study was to determine the prognostic role of elevated levels of chromogranin A (CgA) in terms of biochemical prostate-specific antigen (PSA) progression after radical prostatectomy (RRP) for prostate adenocarcinoma. Two hundred and sixty-four consecutive men with non-metastatic prostate adenocarcinoma submitted to RRP represented our population. In all cases, a blood sample for the determination of serum total PSA and CgA levels was obtained (RIA). Two different upper reference values for serum CgA levels were used: > 60 and > 90 ng/ml. The main end point of this study was biochemical (PSA) progression-free survival. In our population, 35.0% (91/264 cases) of cases presented a serum CgA level > 60 ng/ml and only 6.4% (17/264) presented CgA > 90 ng/ml. After RRP, during a mean follow-up of 64.59 +/- 26.34 months (median 60 months; range 12-120 months), 59 patients (22.3%) showed a biochemical (PSA) progression. Using 60 ng/ml as upper reference value for CgA, 10.4 and 45.0% of cases showed PSA progression after RRP in the group with preoperative CgA levels < or = 60 and > 60 ng/ml respectively. The proportion of PSA progression-free survival was significantly lower in cases with preoperative CgA > 60 ng/ml than in cases with CgA < or = 60 ng/ml (P < 0.0001). In addition, at the multivariate analysis, preoperative serum CgA levels were confirmed as an independent prognostic factor for PSA progression after RRP. In non-metastatic prostate carcinomas, we described a significant prognostic role of CgA in terms of biochemical progression-free survival. PMID- 17914094 TI - The radiation response of hormone-resistant prostate cancer induced by long-term hormone therapy. AB - Hormone therapy for prostate cancer eventually fails leading to a stage called hormone-resistant (HR) disease. To investigate the issue about the characteristics and the radiation response in HR prostate cancer, we established HR cell sub-lines, 22RV1-F and 22RV1-DF, from 22RV1 cells with androgen deprivation for 16 weeks, and obtained LNCaP-HR from LNCaP with long-term bicalutamide treatment. We examined their sensitivities to radiation therapy and the underlying mechanisms. In vitro and in vivo faster tumor growth rate was noted in the HR prostate cancer cells when compared with control. Moreover, HR prostate cancer cells had greater capacity to scavenge reactive oxygen species, and suffered less apoptosis and senescence, and subsequently were more likely to survive from irradiation as measured by clonogenic assay in vitro and growth delay in vivo. The decreased p53 and increased mouse double minute 2 oncogene (MDM2) might be the potential underlying mechanisms for the more aggressive growth and more radioresistance in HR prostate cancer cells. In conclusion, HR prostate cancer cells appeared to be more aggressive in tumor growth and in resistance to radiation treatment. Regulation of the expressions of p53 and MDM2 should be the promising treatment strategies for relative radioresistant prostate cancer. PMID- 17914095 TI - Cytochrome P450 3A5 is highly expressed in normal prostate cells but absent in prostate cancer. AB - Testosterone is essential for the growth and function of the luminal prostate cells, but it is also critical for the development of prostate cancer, which in the majority of the cases derives from luminal cells. Cytochrome P450 3A (CYP3A) enzymes hydroxylate testosterone and dehydroepiandrosterone to less active metabolites, which might be the basis for the association between CYP3A polymorphisms and prostate cancer. However, it is unknown whether the CYP3A enzymes are expressed at relevant levels in the prostate and which polymorphisms could affect this tissue-specific CYP3A activity. Thus, we measured CYP3A4, CYP3A5, CYP3A7, and CYP3A43 mRNA in 14 benign prostatic hyperplasias and ten matched non-tumoral/tumoral prostate samples. We found that CYP3A5 mRNA in non tumoral prostate tissue was 10% of the average amount of liver samples, whereas the expression of the other CYP3A genes was much lower. Similarly to liver, CYP3A5*3 polymorphism decreased CYP3A5 mRNA content 13-fold. CYP3A5 protein was detected in non-tumoral prostate microsomes by western blot, and immunohistochemistry (IHC) localized CYP3A5 exclusively in the basolateral prostate cells. In contrast to the normal tissue, IHC and RT-PCR showed that tumoral tissue lacked CYP3A5 expression. In conclusion, prostate basolateral cells express high levels of CYP3A5 which dramatically decrease in tumoral tissue. This finding supports an endogenous function of CYP3A5 related to the metabolism of intra-prostatic androgens and cell growth, and that polymorphisms affecting CYP3A5 activity may result in altered prostate cancer risk and aggressiveness. PMID- 17914096 TI - HER-2/neu transcriptionally activates Jab1 expression via the AKT/beta-catenin pathway in breast cancer cells. AB - Jab1 is a co-activator of activating protein-1 (AP-1) transcription factor and the fifth subunit of the constitutive photomorphogenesis 9 (COP9) signalosome, which has been shown to mediate nuclear exportation and ubiquitin-dependent degradation of the tumor suppressor p27(Kip1). Jab1 is overexpressed in several types of human cancer. However, de-regulation of Jab1 gene expression in cancer cells is largely unclear. In this study, we reported that expression of Jab1 was stimulated by HER-2/neu oncogene via transcriptional activation. Promoter deletion and mutation analysis indicated that HER-2/neu stimulated Jab1 via the T cell factor (TCF) binding site located at the -380/-368 region of the human Jab1 promoter. DNA affinity precipitation assay and chromatin immunoprecipitation assay verified that binding of beta-catenin and TCF-4 to this consensus site was increased by HER-2/neu. In addition, dominant-negative mutant of TCF significantly attenuated the stimulatory effect of HER-2/neu. We also demonstrated that HER-2/neu increased beta-catenin/TCF-mediated Jab1 expression via the AKT signaling pathway because chemical inhibitor or dominant-negative mutant of AKT effectively attenuated the stimulatory action of HER-2/neu. IGF-I, which is a well-known AKT activator, also up-regulated the expression of Jab1 in NIH/3T3 and MCF-7 cells. Knockdown of Jab1 by small interfering RNA (siRNA) preferentially inhibited proliferation of HER-2/neu-overexpressing breast cancer cells. Taken together, our results suggest that HER-2/neu transcriptionally activates Jab1 expression to promote proliferation of breast cancer cells. PMID- 17914097 TI - Anthropometric measures, plasma adiponectin, and breast cancer risk. AB - Adiponectin is a peptide hormone secreted exclusively by adipocytes, and obesity is an established risk factor for breast cancer. We have, thus, evaluated the associations of anthropometric measures of adiposity and adiponectin with the development of breast cancer in a case-control study. Questionnaire information, anthropometric measures, and blood samples were taken before treatment from 244 incident cases with breast cancer, including 141 premenopausal and 103 postmenopausal cases, and 244 controls admitted for health examination at the Tri Service General Hospital, Taipei between 2004 and 2005. Plasma levels of adiponectin were measured by RIA. The relationship between anthropometric measures of adiposity and breast cancer risk was modified by menopausal status, with a significant increase in risk observed in postmenopausal but not premenopausal women. Moreover, a fairly robust inverse association of adiponectin with the risk was observed only in postmenopausal women (adjusted odds ratio (OR), 0.55; 95% confidence interval (CI), 0.23-0.97), but not in premenopausal women. Additionally, the plasma adiponectin levels tended to be inversely associated with estrogen receptor (ER)-positive (adjusted OR, 0.53; 95% CI, 0.27 0.98) but not ER-negative breast tumors. Furthermore, the associations of adiponectin with breast cancer risk overall and by menopausal and ER status remained after adjustment for obesity indices. These results suggest that adiponectin may have an independent role in breast carcinogenesis, particularly in the postmenopausal and ER-positive breast cancer risk. PMID- 17914098 TI - Activin and estrogen crosstalk regulates transcription in human breast cancer cells. AB - Activin is a member of the transforming growth factor beta superfamily that regulates mammary cell function during development, lactation, and in cancer. Activin slows the growth of breast cancer cells by inducing G(0)/G(1) cell cycle arrest. Estrogen is a steroid hormone that stimulates the proliferation of mammary epithelial cells in development and oncogenesis. The crosstalk between estrogen and activin that regulates activin ligand expression, activin and estrogen signal transduction, and cell cycle arrest was investigated in this study. Estrogen antagonized activin-dependent production of plasminogen activator inhibitor 1 (PAI-1) mRNA, while activin repressed estrogen-dependent transcription of trefoil factor 1. The repression of estrogen signaling by activin was recapitulated using a simple estrogen response element-luciferase construct and was enhanced in the presence of overexpressed estrogen receptor alpha (ERalpha). In contrast, estrogen-mediated repression of activin signaling could not be recapitulated on a simple CAGA Smad-binding element but did inhibit the short PAI-1 promoter, p3TP-luciferase, especially when ERalpha was overexpressed. Repression of both estrogen- and activin-regulated transcription was found to be ligand induced and Smad3 dependent. In addition to transcriptional repression, estrogen also reduced the amount of activin B mRNA and protein produced by MCF7 breast cancer cells. These studies demonstrate the importance of activin and estrogen crosstalk during mammary cell growth and cancer initiation. PMID- 17914099 TI - High tumoral levels of Kiss1 and G-protein-coupled receptor 54 expression are correlated with poor prognosis of estrogen receptor-positive breast tumors. AB - KiSS1 is a putative metastasis suppressor gene in melanoma and breast cancer encoding kisspeptins, which are also described as neuroendocrine regulators of the gonadotropic axis. Negative as well as positive regulation of KiSS1 gene expression by estradiol (E(2)) has been reported in the hypothalamus. Estrogen receptor alpha (ERalpha level is recognized as a marker of breast cancer, raising the question of whether expression of KiSS1 and its G-protein-coupled receptor (GPR54) is down- or upregulated by estrogens in breast cancer cells. KiSS1 was found to be expressed in MDA-MB-231, MCF7, and T47D cell lines, but not in ZR75 1, L56Br, and MDA-MB-435 cells. KiSS1 mRNA levels decreased significantly in ERalpha-negative MDA-MB-231 cells expressing recombinant ERalpha. In contrast, tamoxifen (TAM) treatment of ERalpha-positive MCF7 and T47D cells increased KiSS1 and GPR54 levels. The clinical relevance of this negative regulation of KiSS1 and GPR54 by E(2) was then studied in postmenopausal breast cancers. KiSS1 mRNA increased with the grade of the breast tumors. ERalpha-positive invasive primary tumors expressed sevenfold lower KiSS1 levels than ERalpha-negative tumors. Among ERalpha-positive breast tumors from postmenopausal women treated with TAM, high KiSS1 combined with high GPR54 mRNA tumoral levels was unexpectedly associated with shorter relapse-free survival (RFS) relative to tumors expressing low tumoral mRNA levels of both genes. The contradictory observation of putative metastasis inhibitor role of kisspeptins and RFS to TAM treatment suggests that evaluation of KiSS1 and its receptor tumoral mRNA levels could be new interesting markers of the tumoral resistance to anti-estrogen treatment. PMID- 17914100 TI - Risks of breast, endometrial, and ovarian cancers after twin births. AB - The concentrations of endogenous hormones differ between women with twin and singleton births, with a possible influence on the risk of cancer. We used the nationwide Swedish Family-Cancer Database, including 30,409 women with a twin birth, to examine the subsequent risks of breast, endometrial, and ovarian cancers. Relative risks (RRs) were calculated in a log-linear Poisson regression model of person-years as offset. Cancer data were retrieved from the Swedish Cancer Registry; a total of 1010, 210, and 174 women were diagnosed with breast, endometrial, and ovarian cancers respectively, after a twin birth. A significant decrease in the risk of breast cancer was noted among women with a twin birth compared with women with a singleton birth (RR 0.85, 95% confidence interval (CI) 0.74-0.98). The protective effects were observed throughout the intervals after last pregnancy and they were strongest shortly after the last pregnancy in women who delivered a twin birth before 30 years of age. Twin birth did not change the risk of endometrial cancer (1.08, 95% CI 0.79-1.47) but the RR was increased for women with the number of pregnancies > or =4 (1.39, 95% CI 1.11-1.76). The RR for ovarian cancer was 0.95 (95% CI 0.79-1.15). Our study showed that twin births significantly reduced the subsequent risk of breast cancer. However, the associations of twin births with endometrial and ovarian cancers were not substantial. PMID- 17914101 TI - Human adiponectin inhibits cell growth and induces apoptosis in human endometrial carcinoma cells, HEC-1-A and RL95 2. AB - Obesity is one of the well-established risk factors for endometrial cancer. Recent clinical studies have demonstrated that circulating adiponectin concentrations are inversely correlated with the incidence of endometrial carcinoma. Such epidemiological findings are consistent with the paradoxical observations that adiponectin levels are reduced in obesity. This study investigated the direct effects of adiponectin on two endometrial carcinoma cell lines, HEC-1-A and RL95-2. These cell lines express both variants of adiponectin receptors, adipo-R1 and adipo-R2. Adiponectin treatment leads to suppression of cell proliferation in both cell types, which is primarily due to the significant increase of cell populations at G(1)/G(0)-phase and to the induction of apoptosis. The inhibition of growth in these two cell lines appears to be mediated by different signaling pathways. Although adiponectin treatment markedly increases the phosphorylation (Thr172) of AMP-activated protein kinase alpha in both HEC-1-A and RL95-2 within 30 min, prolonged exposure (48 h) leads to inactivation of Akt as well as reduction of cyclin D1 protein expression in HEC-1 A cells. In contrast, similar treatment of RL95-2 cells with adiponectin, while having no effects on Akt activity and cyclin D1 expression, causes a decrease in cyclin E2 expression and the activity of mitogen-activated kinase (p42/44). We conclude that adiponectin exerts direct anti-proliferative effects on HEC-1-A and RL95-2 cells by inducing cell cycle arrest and apoptosis. Depending on the genotypes of the endometrial cancer cells, the inhibitory effects of adiponectin are associated with the reduction of different pro-growth regulators of cell cycle and signaling proteins. Our study thus provides a cellular mechanism underlying the linkages between endometrial cancer and obesity. PMID- 17914102 TI - Body shape throughout life and correlations with IGFs and GH. AB - Both insulin-like growth factors (IGF) and body size have been linked to premenopausal breast cancer risk. However, observational studies of IGF have not been consistent, and they suggest that perhaps earlier levels of IGF might be more strongly related to breast cancer than those measured at mid-age. We therefore sought to explore associations between several measures of body size throughout life and IGF levels in premenopausal women. We examined cross sectional associations of birth weight, body shape (or somatotype) at ages 5 and 10, body mass index (BMI) at age 18 and adulthood, bra cup size at age 20, adult waist circumference and waist-to-hip ratio (WHR), and attained height with plasma levels of IGF-I, IGF binding protein 3 (IGFBP-3), IGFBP-1, and GH. Participants were 592 healthy premenopausal women aged 34-52 from the Nurses' Health Study II. Using multiple linear regression, we computed least-square mean hormone levels across the categories of early life anthropometric factors. We observed consistent and strong inverse associations between body shape at various stages in life and IGF levels. Somatotype at ages 5 and 10 was inversely associated with IGF-I (P for difference, < 0.01) and positively with IGFBP-3 measured later in adulthood. Further, comparing women with a BMI > or = 25 kg/m(2) at age 18 vs < 19 kg/m(2), similar associations were observed for IGF-I (P for trend, 0.005) and IGFBP-3 (P for trend, 0.01), which were even stronger for BMI at blood collection (BMI< 20 versus BMI > or = 30, mean IGF-I 254 ng/ml, 95% CI, 239-271 vs 208 ng/ml, 95% CI, 195-222). Both waist circumference and WHR were strongly and inversely related to IGFBP-1 levels (top versus bottom quartile of waist circumference: 14.5 vs 40.0 ng/ml, P for trend 0.0005; WHR: 18.3 vs 39.4 ng/ml, P for trend 0.002), with similar results for bra cup size at age 20 although they did not reach statistical significance. There was no association between height and IGF or GH levels. Birth weight, on the other hand, was weakly positively associated with both IGF-I and IGFBP-1 levels, and inversely with GH. Our results suggest that childhood and adult body size may affect premenopausal breast cancer risk differently than birth weight, through associations with IGF and GH levels. PMID- 17914103 TI - Insulin pathway related genes and risk of colorectal cancer: INSR promoter polymorphism shows a protective effect. AB - Western lifestyle leading to obesity and type 2 diabetes has been associated with increased risk of colorectal cancer (CRC). Diet and related factors may affect the risk by modifying plasma insulin levels. Thus, the inter-individual variation in insulin signaling may play a plausible role in the development of CRC. We hypothesized that functional polymorphisms in the insulin pathway genes INS, INSR, IGFBPI, insulin receptor substrate 1 (IRS1), and IRS2 may be associated with CRC. We studied the association of five single nucleotide polymorphisms (SNPs) with the risk of CRC using a hospital-based case-control design with 712 cases and 748 controls from the Czech Republic. The INSR A-603G promoter SNP, which is located within a known Sp1-binding site, was associated with the risk of CRC, with carriers of the G allele having a decreased risk (odds ratios (OR) 0.71, 95% confidence interval (CI) 0.54-0.93). Carrying the variant allele of the IRS1 Gly972Arg SNP further decreased the risk among the INSR-603G allele carriers (OR 0.28, 95% CI 0.11-0.70). SNPs in the INS, IGFBPI, and IRS2 genes did not affect the risk of CRC. In conclusion, genetic variation in the insulin signaling pathway genes may affect the risk of CRC. PMID- 17914104 TI - Estrogenically regulated LRP16 interacts with estrogen receptor alpha and enhances the receptor's transcriptional activity. AB - Previous studies have shown that leukemia related protein 16 (LRP16) is estrogenically regulated and that it can stimulate the proliferation of MCF-7 breast cancer cells, but there are no data on the mechanism of this pathway. Here, we demonstrate that the LRP16 expression is estrogen dependent in several epithelium-derived tumor cells. In addition, the suppression of the endogenous LRP16 in estrogen receptor alpha (ERalpha)-positive MCF-7 cells not only inhibits cells growth, but also significantly attenuates the cell line's estrogen responsive proliferation ability. However, ectopic expression of LRP16 in ERalpha negative MDA-MB-231 cells has no effect on proliferation. These data suggest the involvement of LRP16 in estrogen signaling. We also provide novel evidence by both ectopic expression and small interfering RNA knockdown approaches that LRP16 enhances ERalpha-mediated transcription activity. In stably LRP16-inhibitory MCF 7 cells, the estrogen-induced upregulation of several well-known ERalpha target genes including cyclin D1 and c-myc is obviously impaired. Results from glutathione S-transferase pull-down and coimmunoprecipitation assays revealed that LRP16 physically interacts with ERalpha in a manner that is estrogen independent but is enhanced by estrogen. Furthermore, a mammalian two-hybrid assay indicated that the binding region of LRP16 localizes to the A/B activation function 1 domain of ERalpha. Taken together, these results present new data supporting a role for estrogenically regulated LRP16 as an ERalpha coactivator, providing a positive feedback regulatory loop for ERalpha signal transduction. PMID- 17914105 TI - Metabolic syndrome, plasma lipid, lipoprotein and glucose levels, and endometrial cancer risk in the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - To clarify the role of metabolic factors in endometrial carcinogenesis, we conducted a case-control study nested within the European Prospective Investigation into Cancer and Nutrition (EPIC), and examined the relation between prediagnostic plasma lipids, lipoproteins, and glucose, the metabolic syndrome (MetS; a cluster of metabolic factors) and endometrial cancer risk. Among pre- and postmenopausal women, 284 women developed endometrial cancer during follow up. Using risk set sampling, 546 matched control subjects were selected. From conditional logistic regression models, high-density lipoprotein cholesterol (HDL C) levels were inversely associated with risk body mass index (BMI)-adjusted relative risk (RR) for top versus bottom quartile 0.61 (95% confidence intervals (CI) 0.38-0.97), P(trend) = 0.02). Glucose levels were positively associated with risk (BMI-adjusted RR top versus bottom quartile 1.69 (95% CI 0.99-2.90), P(trend) = 0.03), which appeared stronger among postmenopausal women (BMI adjusted RR top versus bottom tertile 2.61 (95% CI 1.46-4.66), P(trend) = 0.0006, P(heterogeneity) = 0.13) and never-users of exogenous hormones (P(heterogeneity) = 0.005 for oral contraceptive (OC) use and 0.05 for hormone replacement therapy use). The associations of HDL-C and glucose with risk were no longer statistically significant after further adjustment for obesity-related hormones. Plasma total cholesterol, Low-density lipoprotein cholesterol (LDL-C), and triglycerides were not significantly related to overall risk. The presence of MetS was associated with risk (RR 2.12 (95% CI 1.51-2.97)), which increased with the number of MetS factors (P(trend) = 0.02). An increasing number of MetS factors other than waist circumference, however, was marginally significantly associated with risk only in women with waist circumference above the median (P(interaction) = 0.01). None of the associations differed significantly by fasting status. These findings suggest that metabolic abnormalities and obesity may act synergistically to increase endometrial cancer risk. PMID- 17914106 TI - DNA copy number status is a powerful predictor of poor survival in endocrine pancreatic tumor patients. AB - The clinical behavior of endocrine pancreatic tumors (EPTs) is difficult to predict in the absence of metastases or invasion to adjacent organs. Several markers have been indicated as potential predictors of metastatic disease, such as tumor size > or =2 cm, Ki67 proliferative index > or =2%, cytokeratin (CK) 19 status, and recently in insulinomas, chromosomal instability (CIN). The goal of this study was to evaluate the value of these markers, and in particular of the CIN, to predict tumor recurrence or progression and tumor-specific death, using a series of 47 insulinomas and 24 non-insulinoma EPTs. From these EPT cases, a genomic profile has been generated and follow-up data have been obtained. The proliferative index has been determined in 68 tumors and a CK19 expression pattern in 50 tumors. Results are statistically analyzed using Kaplan-Meier plots and the log-rank statistic. General CIN, as well as specific chromosomal alterations such as 3p and 6q loss and 12q gain, turned out to be the most powerful indicators for poor tumor-free survival (P< or =0.0004) and tumor specific death (P< or =0.0113) in insulinomas. The CIN, chromosome 7q gain, and a proliferative index > or =2% were reliable in predicting a poor tumor-free survival in non-insulinoma EPTs (P< or =0.0181, whereas CK19 expression was the most optimal predictor of tumor-specific death in these tumors. In conclusion, DNA copy number status is the most sensitive and efficient marker of adverse clinical outcome in insulinomas and of potential interest in non-insulinoma EPTs. As a consequence, this marker should be considered as a prognosticator to improve clinical diagnosis, most practically as a simple multi-target test. PMID- 17914107 TI - IGF axis gene expression patterns are prognostic of survival in epithelial ovarian cancer. AB - The IGF axis has documented growth-promoting effects in various malignancies, but its role in epithelial ovarian cancer (EOC) has not been adequately examined. We studied the expression of the IGF axis genes in relation to outcome in EOC. Microarray expression profiles from 64 patients with advanced-stage EOC were used. Two multi-gene subsets were chosen, one upstream of the IGF receptor ('IGF family') and the other downstream of the IGF receptor ('IGF signaling pathway'), and analyzed in relation to survival. In addition, expression patterns of the two gene subsets were analyzed in relation to favorable and unfavorable prognosis categories identified in a previous study by whole-genome expression profiling. In a gene-by-gene analysis, IGF binding protein 4 and IGF-II receptor gene expression was inversely associated with survival. Using hierarchical clustering, the two multi-gene subsets separated the patient cohort into two groups with different median survival (IGF family: 33 vs 63 months, P=0.02 and IGF signaling pathway: 41 vs 63 months, P=0.05). Furthermore, the two multi-gene subsets were differentially expressed between the previously defined favorable and unfavorable prognosis tumors (Kolmogorov-Smirnov permutation: P=0.0005 and 0.003 for the IGF family and signaling pathway respectively), and individual genes (including IGF I, IGF-I receptor, and several genes downstream of the receptor) were overexpressed in unfavorable prognosis tumors (permutation P<0.05). The expression patterns of several genes in the IGF axis are associated with survival in EOC, and expression changes of these genes may be underlying previously proposed microarray-derived clinical prognostic models. Future studies are needed to more precisely determine the diagnostic and potential therapeutic significance of these findings. PMID- 17914108 TI - MicroRNAs (miR)-221 and miR-222, both overexpressed in human thyroid papillary carcinomas, regulate p27Kip1 protein levels and cell cycle. AB - We have recently reported that MicroRNAs (miR)-221 and miR-222 were up-regulated in human thyroid papillary carcinomas in comparison with the normal thyroid tissue. Bioinformatic analysis proposed the p27(Kip1) protein, a key regulator of cell cycle, as a candidate target for the miR-221/222 cluster. Here, we report that the enforced expression of miR-221 and miR-222 was able to reduce p27(Kip1) protein levels in thyroid carcinoma and HeLa cells in the absence of significant changes in specific p27(Kip1) mRNA levels. This effect is direct as miR-221 and miR-222 negatively regulate the expression of the 3'-untranslated region-based reporter construct from the p27(Kip1) gene, and is dependent on two target sites in this region. Consistent with these results, an enforced expression of the miR 221 and miR-222 induced the thyroid papillary carcinoma cell line (TPC-1) to progress to the S phase of the cell cycle. It is likely that the negative regulation of p27(Kip1) by miR-221 and miR-222 might also have a role in vivo since we report an inverse correlation between miR-221 and miR-222 up-regulation and down-regulation of the p27(Kip1) protein levels in human thyroid papillary carcinomas. Therefore, the data reported here demonstrate that miR-221 and miR 222 are endogenous regulators of p27(Kip1) protein expression, and thereby, the cell cycle. PMID- 17914109 TI - Bone metastases of differentiated thyroid cancer: impact of early 131I-based detection on outcome. AB - Bone is the second most frequent target of distant metastases in patients with differentiated thyroid cancer, and such forms carry a very poor prognosis. The impact of (131)I therapy in this setting is controversial. We describe the diagnostic circumstances and outcome of patients with bone metastases recently managed in two institutions. Among 921 consecutive thyroid cancer patients who had total thyroidectomy and (131)I ablation between January 2000 and December 2004 and who were subsequently monitored, bone metastases had been diagnosed in 16 patients. In three cases, the bone metastases were non-functioning (negative (131)I uptake) . These patients were treated with surgery and radiotherapy but progressed rapidly. The other 13 patients had functioning (positive (131)I uptake) bone metastases. In five of them, thyroid cancer was revealed by signs of distant involvement (bone pain, n = 4; dyspnea, n = 1). The bone metastases progressed in these five patients, despite local therapy and multiple courses of (131)I. The bone metastases in the remaining eight patients were discovered on the post-surgery (131)I therapy scan. Complementary radiological studies were negative except in one patient in whom one of the metastases (a 5 mm lesion of the right humerus) was visible on magnetic resonance imaging (MRI). Six of these patients showed a good response to (131)I therapy, with (131)I uptake and Tg levels becoming undetectable or showing a sharp fall. One patient refused (131)I therapy; bone metastases became visible on MRI within 1 year and the Tg level rose tenfold. The disease progressed in one patient despite (131)I therapy. Post surgical (131)I ablation can contribute to early detection of bone metastases at a time when the Tg level may be only moderately elevated, when other radiological studies are negative, and when the disease is potentially curable by (131)I therapy. PMID- 17914110 TI - A multi-gene approach to differentiate papillary thyroid carcinoma from benign lesions: gene selection using support vector machines with bootstrapping. AB - Selection of novel molecular markers is an important goal of cancer genomics studies. The aim of our analysis was to apply the multivariate bioinformatical tools to rank the genes - potential markers of papillary thyroid cancer (PTC) according to their diagnostic usefulness. We also assessed the accuracy of benign/malignant classification, based on gene expression profiling, for PTC. We analyzed a 180-array dataset (90 HG-U95A and 90 HG-U133A oligonucleotide arrays), which included a collection of 57 PTCs, 61 benign thyroid tumors, and 62 apparently normal tissues. Gene selection was carried out by the support vector machines method with bootstrapping, which allowed us 1) ranking the genes that were most important for classification quality and appeared most frequently in the classifiers (bootstrap-based feature ranking, BBFR); 2) ranking the samples, and thus detecting cases that were most difficult to classify (bootstrap-based outlier detection). The accuracy of PTC diagnosis was 98.5% for a 20-gene classifier, its 95% confidence interval (CI) was 95.9-100%, with the lower limit of CI exceeding 95% already for five genes. Only 5 of 180 samples (2.8%) were misclassified in more than 10% of bootstrap iterations. We specified 43 genes which are most suitable as molecular markers of PTC, among them some well-known PTC markers (MET, fibronectin 1, dipeptidylpeptidase 4, or adenosine A1 receptor) and potential new ones (UDP-galactose-4-epimerase, cadherin 16, gap junction protein 3, sushi, nidogen, and EGF-like domains 1, inhibitor of DNA binding 3, RUNX1, leiomodin 1, F-box protein 9, and tripartite motif-containing 58). The highest ranking gene, metallophosphoesterase domain-containing protein 2, achieved 96.7% of the maximum BBFR score. PMID- 17914112 TI - Valproic acid enhances tubulin acetylation and apoptotic activity of paclitaxel on anaplastic thyroid cancer cell lines. AB - The introduction of paclitaxel into multimodal therapy for anaplastic thyroid carcinoma has failed to improve overall survival. Toxicity rules out the high doses required, especially in older patients. The search for strategies to enhance paclitaxel antineoplastic activity and reduce its side effects is thus advisable. The study aimed to determine whether the histone deacetylase (HDAC) inhibitor valproic acid (VPA) improves the anticancer action of paclitaxel and elucidate the mechanisms underlying the effects of combined treatment. We examined the effect of VPA on the sensitivity to paclitaxel of two anaplastic thyroid carcinoma cell lines (CAL-62 and ARO), and the ability of the drug to determine tubulin acetylation and enhance paclitaxel-induced acetylation. The addition of as little as 0.7 mM VPA to paclitaxel enhances both cytostatic and cytotoxic effects of paclitaxel alone. Increased apoptosis explains the enhancement of the cytotoxic effect. The mechanism underlying this effect is through inhibition of HDAC6 activity, which leads to tubulin hyperacetylation. The results suggest a mechanistic link between HDAC6 inhibition, tubulin acetylation, and the VPA-induced enhancement of paclitaxel effects, and provide the rationale for designing future combination therapies. PMID- 17914111 TI - Transforming acidic coiled-coil 3 and Aurora-A interact in human thyrocytes and their expression is deregulated in thyroid cancer tissues. AB - Aurora-A kinase has recently been shown to be deregulated in thyroid cancer cells and tissues. Among the Aurora-A substrates identified, transforming acidic coiled coil (TACC3), a member of the TACC family, plays an important role in cell cycle progression and alterations of its expression occur in different cancer tissues. In this study, we demonstrated the expression of the TACC3 gene in normal human thyroid cells (HTU5), and its modulation at both mRNA and protein levels during cell cycle. Its expression was found, with respect to HTU5 cells, unchanged in cells derived from a benign thyroid follicular tumor (HTU42), and significantly reduced in cell lines derived from follicular (FTC-133), papillary (B-CPAP), and anaplastic thyroid carcinomas (CAL-62 and 8305C). Moreover, in 16 differentiated thyroid cancer tissues, TACC3 mRNA levels were found, with respect to normal matched tissues, reduced by twofold in 56% of cases and increased by twofold in 44% of cases. In the same tissues, a correlation between the expression of the TACC3 and Aurora-A mRNAs was observed. TACC3 and Aurora-A interact in vivo in thyroid cells and both proteins localized onto the mitotic structure of thyroid cells. Finally, TACC3 localization on spindle microtubule was no more observed following the inhibition of Aurora kinase activity by VX-680. We propose that Aurora-A and TACC3 interaction is important to control the mitotic spindle organization required for proper chromosome segregation. PMID- 17914113 TI - Selective embolization of thyroid arteries as a preresective and palliative treatment of thyroid cancer. AB - Although many tumours of head and neck have been successfully embolized, the number of publications on the application of selective embolization of thyroid arteries (SETA) is limited. The aim of the present study is to evaluate the safety, efficacy and possible indications and contraindications for preresective or palliative SETA in thyroid cancer. The study group comprised 20 patients with thyroid tumours: 7 cases of advanced inoperable anaplastic thyroid cancer (ATC) and 13 cases of differentiated thyroid carcinoma (DTC). All the patients underwent SETA of the superior and/or inferior thyroid arteries. After SETA, selective angiographies of thyroid arteries were performed to ensure that the targeted arteries had been completely occluded. In all the cases, SETA decreased the blood flow through the thyroid. Preresective SETA limited bleeding during surgery and decreased operating time. We observed a massive increase of thyroglobulin (Tg) concentrations in cases of DTC that started 36-48 h after SETA and did not occur in cases of ATC. Although SETA had no influence on the mortality of ATC patients, they reported improvements in swallowing, breathing and decrease of the pain. Concluding, SETA is minimally invasive and safe method limiting blood flow through thyroid tumours. In DTC patients, SETA causes ischaemic necrosis of the gland which results in important increases in serum concentrations of Tg. Therefore, thyroidectomy should be performed during the first 36 h after preresective embolization. Moreover, SETA may become an attractive option of palliative treatment for ATC patients with intractable bleeding, pain or signs of tracheal and oesophageal compression. PMID- 17914114 TI - Gastrointestinal stromal tumors regularly express synaptic vesicle proteins: evidence of a neuroendocrine phenotype. AB - Gastrointestinal stromal tumors (GISTs) are thought to originate from the interstitial cells of Cajal, which share many properties with neurons of the gastrointestinal tract. Recently, we demonstrated expression of the hormone ghrelin in GIST. The aim of the present study was therefore to evaluate a possible neuroendocrine phenotype of GIST. Specimens from 41 GISTs were examined for the expression of 12 different synaptic vesicle proteins. Expression of synaptic-like microvesicle proteins, e.g., Synaptic vesicle protein 2 (SV2), synaptobrevin, synapsin 1, and amphiphysin was demonstrated in a majority of GISTs by immunohistochemistry, western blotting, and quantitative reversetranscriptase PCR. One-third of the tumors also expressed the large dense core vesicle protein vesicular monoamine transporter 1. Presence of microvesicles and dense core vesicles in GIST was confirmed by electron microscopy. The expression of synaptic-like microvesicle proteins in GIST was not related to risk profile or to KIT/platelet derived growth factor alpha (PDGFRA) mutational status. Thus, GISTs regularly express a subset of synaptic-like microvesicle proteins necessary for the regulated secretion of neurotransmitters and hormones. Expression of synaptic-like micro-vesicle proteins, ghrelin and peptide hormone receptors in GIST indicate a neuroendocrine phenotype and suggest novel possibilities to treat therapy-resistant GIST. PMID- 17914115 TI - Snail and Sonic Hedgehog activation in neuroendocrine tumors of the ileum. AB - The transcription factor Snail represses E-cadherin and induces epithelial mesenchymal transition, a process also exploited by invasive cancer cells. Aberrant Hedgehog (Hh) signaling was recently observed in a variety of epithelial cancers and it has been shown that the Hh target gene Gli1 induces expression of Snail. In this study, we examined whether Snail and Sonic Hedgehog (SHH) are expressed in neuroendocrine tumors (NETs) of the ileum. Using immunohistochemistry, we found expression of Snail in 22 out of 37 (59%) of evaluated NET samples, but not in adjacent normal tissues. Snail expression was mostly restricted to the invasive front of the tumors. Six of seven liver metastases analyzed were positive for Snail. Intratumoral expression of SHH was detected in 27 out of 37 (73%) tumors. As opposed to Snail, cells expressing SHH were found to be distributed more randomly throughout the tumors. Out of 30 primary NETs, 16 (53%) showed both Snail and SHH expression. Furthermore, we found downregulation of E-cadherin in Snail-expressing cells by immunofluorescence. Real-time RT-PCR revealed conservation of the Hh target genes Gli1, Gli2, and Ptch in the pancreatic carcinoid cell line BON-1, which were downregulated upon Hh inhibition with cyclopamine. Moreover, Hh inhibition attenuated in vitro cell growth in a dose-dependent manner. In conclusion, we describe for the first time that Snail and SHH are overexpressed in a large subset of NETs of the ileum. Aberrant activation of these pathways might be involved in invasion and metastatic spread in NETs. PMID- 17914116 TI - The Mia/Cd-rap gene expression is downregulated by the high-mobility group A proteins in mouse pituitary adenomas. AB - The high-mobility group A (HMGA) family of proteins orchestrates the assembly of nucleoprotein structures playing important roles in gene transcription, recombination, and chromatin structure through a complex network of protein-DNA and protein-protein interactions. Recently, we have generated transgenic mice carrying wild type or truncated HMGA2 genes under the transcriptional control of the cytomegalovirus promoter. These mice developed pituitary adenomas secreting prolactin and GH mainly due to an increased E2F1 activity, directly consequent to the HMGA2 overexpression. To identify other genes involved in the process of pituitary tumorigenesis induced by the HMGA2 gene, in this study we have analyzed the gene expression profile of three HMGA2-pituitary adenomas in comparison with a pool of ten normal pituitary glands from control mice, using the Affymetrix MG MU11K oligonucleotide array representing approximately 13,000 unique genes. We have identified 82 transcripts that increased and 72 transcripts that decreased at least four-fold in all the mice pituitary adenomas analyzed compared with normal pituitary glands. Among these genes, we focused our attention on the Mia/Cd-rap gene, whose expression was essentially suppressed in all of the pituitary adenomas tested by the microarray. We demonstrated that the HMGA proteins directly bind to the promoter of the Mia/Cd-rap gene and are able to downregulate its expression. In order to understand a possible role of Mia/Cd-rap in pituitary cell growth, we performed a colony assay in GH3 and GH4 cells. Interestingly, Mia/Cd-rap expression inhibits their proliferation, suggesting a potential tumor suppressor role of Mia/Cd-rap in pituitary cells. PMID- 17914117 TI - A diagnostic marker set for invasion, proliferation, and aggressiveness of prolactin pituitary tumors. AB - Although most pituitary tumors are benign, some are invasive or aggressive. In the absence of specific markers of malignancy, only tumors with metastases are considered malignant. To identify markers of invasion and aggressiveness, we focused on prolactin (PRL) tumors in the human and rat. Using radiology and histological methods, we classified 25 human PRL tumors into three groups (non invasive, invasive, and aggressive-invasive) and compared them with a model of transplantable rat PRL tumors with benign and malignant lineages. Combining histological(mitoses and labeling for Ki-67, P53, pituitary transforming tumor gene (PTTG), and polysialic acid neural cell adhesion molecule) and transcriptomic (microarrays and q-RTPCR) methods with clinical data (post surgical outcome with case-control statistical analysis), we found nine genes implicated in invasion (ADAMTS6, CRMP1, and DCAMKL3) proliferation (PTTG, ASK, CCNB1, AURKB, and CENPE), or pituitary differentiation (PITX1) showing differential expression in the three groups of tumors (P = 0.015 to 0.0001). A case-control analysis, comparing patients in remission (9 controls) and patients with persistent or recurrent tumors (14 cases) revealed that eight out of the nine genes were differentially up- or downregulated (P = 0.05 to 0.002), with only PTTG showing no correlation with clinical course (P = 0.258). These combined histological and transcriptomic analyses improve the pathological diagnosis of PRL tumors, indicating a reliable procedure for predicting tumor aggressiveness and recurrence potential. The similar gene profiles found between non-invasive human and benign rat tumors, as well as between aggressive-invasive human and malignant rat tumors provide new insights into malignancy in human pituitary tumors. PMID- 17914118 TI - No evidence of somatic aryl hydrocarbon receptor interacting protein mutations in sporadic endocrine neoplasia. AB - Germline mutations in the aryl hydrocarbon receptor interacting protein (AIP) gene were recently observed in patients with pituitary adenoma predisposition (PAP). Though AIP mutation-positive individuals with prolactin-, mixed growth hormone/prolactin-, and ACTH-producing pituitary adenomas as well as non secreting pituitary adenomas have been reported, most mutation-positive patients have had growth hormone-producing adenomas diagnosed at relatively young age. Pituitary adenomas are also component tumors of some familial endocrine neoplasia syndromes such as multiple endocrine neoplasia type 1 (MEN1) and Carney complex (CNC). Genes underlying MEN1 and CNC are rarely mutated in sporadic pituitary adenomas, but more often in other lesions contributing to these two syndromes. Thus far, the occurrence of somatic AIP mutations has not been studied in endocrine tumors other than pituitary adenomas. Here, we have analyzed 32 pituitary adenomas and 79 other tumors of the endocrine system for somatic AIP mutations by direct sequencing. No somatic mutations were identified. However, two out of nine patients with prolactin-producing adenoma were shown to harbor a Finnish founder mutation (Q14X) with a complete loss of the wild-type allele in the tumors. These results are in agreement with previous studies in that prolactin-producing adenomas are component tumors in PAP. The data also support the previous finding that somatic AIP mutations are not common in pituitary adenomas and suggest that such mutations are rare in other endocrine tumors as well. PMID- 17914119 TI - Divergent roles of IFNs in the sensitization to endotoxin shock by lactate dehydrogenase-elevating virus. AB - The effect of mouse infection with lactate dehydrogenase-elevating virus (LDV), a usually non-pathogenic virus, on concomitant bacterial endotoxin shock was analyzed, in terms of lethality and cytokine production. A strong enhancement of susceptibility to the shock was observed in mice acutely infected with this virus. It correlated with a sharp increase of tumor necrosis factor and leukemia inhibitory factor production and was controlled by the mouse genetic background. The viral infection led to an imbalance in the cytokine response to LPS, with an enhancement of pro-inflammatory cytokines, including IL-18 and IFN-gamma and a delayed secretion of anti-inflammatory IL-10 that could result in exacerbated macrophage activation. Enhanced IFN-gamma production was involved in the virus induced susceptibility to shock. In sharp contrast with other viral infections, IFN-alpha/beta diminished IFN-gamma production and the resulting increased response to LPS in LDV-infected animals. PMID- 17914120 TI - Abrogation of CCL21 chemokine function by transgenic over-expression impairs T cell immunity to local infections. AB - The CC chemokine receptor 7 (CCR7) and its two ligands, CCL21 and CCL19, play an important role in migration of immune cells to lymphoid tissue. To analyze the function of CCR7 in T cell immunity to infectious agents in vivo, transgenic (tg) mice expressing CCL21 in an ubiquitous fashion were generated. These mice contained high amounts of CCL21 in the serum ( approximately 0.3 microg/ml that resulted in CCR7 down-regulation and in a strongly impaired migration of T cells toward CCL21 in vitro. Lymph nodes in CCL21-tg mice were reduced in size but with intact microanatomy and normal distribution of T and B cells. CCL21-tg mice showed a significantly decreased CD8 T cell response to lymphocytic choriomeningitis virus after footpad infection, whereas the response after systemic infection was not altered. Likewise, the CD4 T cell response to footpad infection with Leishmania major was considerably lowered and CCL21-tg mice failed to clear parasites from infected skin. Taken together, these data demonstrate the importance of CCR7 in mediating T cell immunity to viral and parasitic pathogens after local infection. PMID- 17914121 TI - Familial aggregation of left main coronary artery disease and future risk of coronary events in asymptomatic siblings of affected patients. AB - AIMS: Recently, we observed in a hypothesis-generating exploratory search on the heritability of coronary morphology that left main coronary disease (LMD) was frequently shared by siblings with coronary artery disease (CAD). Thus, our aims were, first, to test specifically the familial aggregation of LMD and second, to investigate whether LMD is a stronger predictor for future incident events than other manifestations of CAD in seemingly healthy siblings of CAD patients. METHODS AND RESULTS: Coronary angiograms of 1801 patients (n = 882 from the initial exploratory study and 919 additional angiograms) were analysed from families with > or = 2 affected CAD siblings. We estimated the heritability using the variance-component methodology and sibling recurrent risks by logistic regression analysis. Moreover, we studied 1369 healthy siblings of CAD patients with known coronary morphology who had a subsequent coronary event by conducting a prospective, nested case-control study. LMD-frequency was comparable in our initial exploratory study (11%) and the new sample (12%). The heritability of LMD was significant in the exploratory 48%, P = 0.010, in the subsequent 45%, P = 0.045, and in the total study sample 49%, P = 0.002. The sibling recurrent risk ratio to present with LMD was 3.6 [CI 1.7-7.1] when another sibling was affected by LMD. In the prospective study on initially healthy family members of CAD patients, 79 siblings experienced an event during follow-up. LMD was more frequently found in families with an event than in families without (13.9 vs. 6.4%, P = 0.036). The relative risk for initially asymptomatic siblings of patients with LMD to suffer from a coronary event was 2.5 [CI 1.1-5.8] compared with siblings of patients with other manifestations of CAD. CONCLUSION: These data confirm our initial observation of familial aggregation of LMD. Moreover, in apparently healthy siblings of patients with LMD, this heritable component results in a risk increase for future events that is greater than that of a strong positive family history by itself. PMID- 17914122 TI - Clean as a whistle, but what about that wheeze? PMID- 17914123 TI - Breaking down the "Great Wall" of COPD care in China. PMID- 17914124 TI - Powering up failed organs. PMID- 17914125 TI - Dendritic cell maturity and obstructive airway disease. PMID- 17914126 TI - Is giant cell interstitial pneumonitis synonymous with hard metal lung disease? PMID- 17914128 TI - Increased energy expenditure after dilutional exchange transfusion for neonatal polycythemia. AB - OBJECTIVE: Hypothermia is a known symptom of neonatal polycythemia (NP) and its pathophysiology is unclear. The effect of partial dilutional exchange transfusion (PET) upon resting energy expenditure (REE) is unknown. We aimed to test the hypothesis that PET leads to an increase in REE. STUDY DESIGN: 11 patients with NP who underwent PET and 10 controls without polycythemia were studied. NP was defined as a venous HCT >/=0.65. Per protocol, symptomatic infants and/or those with venous HCT > or =0.70 underwent PET. REE was measured just prior and 23 hours after PET in patients with NP and at identical ages in the control group. Infants were studied in a skin servo controlled radiant warmer, while clinically and thermally stable, prone and asleep. Measurements were stopped during body movements (less than 5% of the time of measurement). Metabolic measurements were performed by indirect calorimetry, using the Deltatrac II Metabolic monitor (Datex-Ohmeda, Helsinki, Finland). This instrument uses the principle of the open circuit system that allows continuous measurements of oxygen consumption (Vo(2)) and carbon dioxide production (Vco(2)) using a constant flow generator. REE measurements were corrected for the infant weight (Kcal/kg/d). Comparison of REE values between groups was performed using paired Wilcoxon ranked test. RESULTS: Patients with and without NP had nearly identical baseline REE. In patients with NP, REE increased from 44.0 +/- 6.6 Kcal/Kg/d to 48.3 +/- 5.1 Kcal/Kg/d after PET (P<0.05). Furthermore, the increase in REE following PET correlated inversely with the decrease in hematocrit. There was no significant change in REE over time in the control group. In the NP group, symptomatic infants (n=5) had a significantly greater increase in REE following PET than non-symptomatic ones (1.4 +/- 6.3 vs. 7.8 +/- 4.9 Kcal/Kg/d, p<0.05). CONCLUSIONS: Energy expenditure of polycythemic infants increases following PET, in a manner proportional to the decrease in hematocrit. Symptomatic polycythemic infants have a greater rise in REE following PET than non-symptomatic ones. We speculate that polycythemia leads to a decreased REE that might be remedied by PET. PMID- 17914127 TI - Effect of low dose antioxidant vitamin and trace element supplementation on the urinary concentrations of thromboxane and prostacyclin metabolites. AB - OBJECTIVE: This trial evaluated the effect of antioxidant supplementation on the urinary excretion of 11-dehydro TXB(2)/2,3 dinor 6 keto PGF(1alpha) ratio, a marker of the pathogenesis of thrombosis and arteriosclerosis. METHODS: This study was a randomised, double-blind, placebo-controlled trial involving 186 presumably healthy volunteers. One hundred received a multi-antioxidant supplementation and 86 a placebo for two years. Blood zinc, selenium, beta carotene, vitamin C and E and urinary excretion of 11-dehydro TXB(2) and 2,3 dinor 6 keto PGF(1alpha) were measured. RESULTS: Baseline subject characteristics did not differ between the two groups. Blood zinc, selenium, and beta-carotene concentrations significantly increased between baseline and two years in the multi-antioxidant supplementation group supporting subject compliance (p < 0.05). At two years, the median urinary 11-dehydro TXB(2)/2,3 dinor 6 keto PGF(1alpha) ratio was significantly lower in the multi-antioxidant supplementation group (3.4 versus 2.78, p = 0.015). Serum selenium concentration was the only antioxidant studied that was significantly related to the urinary 11-dehydro TXB(2)/2,3 dinor 6 keto PGF(1alpha) ratio. CONCLUSIONS: These results support the hypothesis that a low-dose multi-antioxidant supplementation may contributes to a reduction in platelet activation which is beneficial for cardiovascular function. PMID- 17914129 TI - Soybean protein hydrolysate improves plasma and liver lipid profiles in rats fed high-cholesterol diet. AB - OBJECTIVE: This investigation attempted to clarify the hypolipidemic effects of non-dialyzed soybean protein hydrolysate (NSPH), which is hydrolyzed by pepsin from soybean acid-precipitated protein (APP), in rats fed a cholesterol-rich diet. METHODS: Forty Sprague-Dawley rats were divided into four groups as the control group (19.7% casein), the APP group (14.7% casein + 5% APP), the NSPH group (14.7% casein + 5% NSPH), and the ISO group (19.7% casein + 0.0013% soy isoflavone). RESULTS: After 12-week experimental period, the APP and NSPH groups had a significant lower plasma total cholesterol, triglycerides, and LDL cholesterol concentrations compared with the control group. Additionally, the atherosclerosis index in APP and NSPH group had also markedly decreased. Liver cholesterol and triglyceride contents of the APP and NSPH group were significantly lower than those of the control group. There were no different in plasma LDL-C, liver cholesterol and triglycerides between the ISO group and control group. Fecal excretion of neutral steroids and nitrogen compounds was significantly higher in the APP and NSPH groups than that in the control group. An in vitro study also showed that NSPH, compared with casein, obviously decreased cholesterol micellar solubility. CONCLUSION: These results suggested that NSPH may decrease lipid accumulation in the liver and have a hypolipidemic effect by enhancing excretion and inhibiting absorption of lipids. PMID- 17914130 TI - Ecologic correlates of obesity in rural obese adults. AB - OBJECTIVE: We examined relationships of individual and environmental factors with obesity and trying to lose weight in rural residents. METHODS: The joint contributions of individual and environmental factors on obesity status (obese vs. morbidly obese) and trying to lose weight (yes vs. no) were evaluated using generalized estimating equations. Patients at 29 clinics in rural areas (N = 414, M age 55.0 years (SD = 15.4), 66.3% female) completed anthropometric assessments of weight and height along with survey assessments of individual sociodemographics and trying to lose weight. Rural environments were assessed on aggregated physician access, and sociodemographic context. RESULTS: Most participants (70%, M BMI = 38.3) were obese and 30% morbidly obese. A majority (73%, n = 302) of the sample was trying to lose weight. Compared to obese, morbidly obese participants were more likely to be younger, disproportionately female, not have private insurance, have more comorbid conditions, and rate themselves in worse health in comparison to their obese peers. Compared to not trying to lose weight, trying to lose weight participants were more likely to be younger, disproportionately female, have fewer comorbid conditions, and have attempted to lose weight more times through exercise. Few relationships were seen between environmental variables and obesity or trying to lose weight. CONCLUSIONS: There was no consistent pattern of relationships between environment factors and obesity or trying to lose weight was seen. Unique aspects of rural living may not be captured by traditionally available neighborhood measures. PMID- 17914131 TI - A MUFA-rich diet improves posprandial glucose, lipid and GLP-1 responses in insulin-resistant subjects. AB - OBJECTIVE: To study the effects of three weight-maintenance diets with different macronutrient composition on carbohydrate, lipid metabolism, insulin and incretin levels in insulin-resistant subjects. METHODS: A prospective study was performed in eleven (7 W, 4 M) offspring of obese and type 2 diabetes patients. Subjects had a BMI > 25 Kg/m2, waist circumference (men/women) > 102/88, HBA1c < 6.5% and were regarded as insulin-resistant after an OGTT (Matsuda ISIm <4). They were randomly divided into three groups and underwent three dietary periods each of 28 days in a crossover design: a) diet high in saturated fat (SAT), b) diet rich in monounsaturated fat (MUFA; Mediterranean diet) and c) diet rich in carbohydrate (CHO). RESULTS: Body weight and resting energy expenditure did not changed during the three dietary periods. Fasting serum glucose concentrations fell during MUFA rich and CHO-rich diets compared with high-SAT diets (5.02 +/- 0.1, 5.03 +/- 0.1, 5.50 +/- 0.2 mmol/L, respectively. Anova < 0.05). The MUFA-rich diet improved insulin sensitivity, as indicated by lower homeostasis model analysis-insulin resistance (HOMA-ir), compared with CHO-rich and high-SAT diets (2.32 +/- 0.3, 2.52 +/- 0.4, 2.72 +/- 0.4, respectively, Anova < 0.01). After a MUFA-rich and high-SAT breakfasts (443 kcal) the postprandial integrated area under curve (AUC) of glucose and insulin were lowered compared with isocaloric CHO-rich breakfast (7.8 +/- 1.3, 5.84 +/- 1.2, 11.9 +/- 2.7 mmol . 180 min/L, Anova < 0.05; and 1004 +/- 147, 1253 +/- 140, 2667 +/- 329 pmol . 180 min/L, Anova <0.01, respectively); while the integrated glucagon-like peptide-1 response increased with MUFA and SAT breakfasts compared with isocaloric CHO-rich meals (4.22 +/- 0.7, 4.34 +/- 1.1, 1.85 +/- 1.1, respectively, Anova < 0.05). Fasting and postprandial HDL cholesterol concentrations rose with MUFA-rich diets, and the AUCs of triacylglycerol fell with the CHO-rich diet. Similarly fasting proinsulin (PI) concentration fell, while stimulated ratio PI/I was not changed by MUFA-rich diet. CONCLUSIONS: Weight maintenance with a MUFA-rich diet improves HOMA-ir and fasting proinsulin levels in insulin-resistant subjects. Ingestion of a virgin olive oil-based breakfast decreased postprandial glucose and insulin concentrations, and increased HDL-C and GLP-1 concentrations as compared with CHO rich diet. PMID- 17914132 TI - Specific formulation of Camellia sinensis prevents cold and flu symptoms and enhances gamma,delta T cell function: a randomized, double-blind, placebo controlled study. AB - OBJECTIVE: Determine if a specific formulation of Camellia sinensis (CSF) can prevent illness and symptoms due to cold and flu, and enhance gammadelta T cell function METHODS: DESIGN: Randomized, double-blind, placebo-controlled study. SUBJECTS: Healthy adults 18-70 years old. INTERVENTION: Proprietary formulation of Camellia sinensis (green tea) capsules, or a placebo, twice a day, for 3 months. MEASURES OF OUTCOME: As assessed by daily symptom logs, percentage of subjects experiencing cold and flu symptoms, number of days subjects experienced symptoms, and percentage of subjects seeking medical treatment. Mean in vivo and ex vivo proliferative and interferon gamma responses of subjects' peripheral blood mononuclear cells to gammadelta T cell antigen stimulation. RESULTS: Among subjects taking CSF there were 32.1% fewer subjects with symptoms (P = 0.035), 22.9% fewer overall illnesses of at least 2 days duration (P = 0.092), and 35.6% fewer symptom days (P < 0.002), compared to subjects taking placebo. gammadelta T cells from subjects taking CSF proliferated 28% more (P = 0.017) and secreted 26% more IFN-gamma (P = 0.046) in response to gammadelta T cell antigens, as compared to gammadelta T cells from subjects taking placebo. CSF was well-tolerated. CONCLUSIONS: This proprietary formulation of CSF is a safe and effective dietary supplement for preventing cold and flu symptoms, and for enhancing gammadelta T cell function. PMID- 17914133 TI - Preliminary evidence shows that folic acid fortification of the food supply is associated with higher methotrexate dosing in patients with rheumatoid arthritis. AB - BACKGROUND: Fortification of the diet with folate has been used in the United States since 1997 to prevent neural tube defects in newborn babies. However, an increase in dietary folate intake could theoretically reduce the effectiveness of the anti-folate medication, methotrexate (MTX) in treating rheumatoid arthritis (RA) and other inflammatory diseases. OBJECTIVE: To investigate whether dietary fortification with folic acid interferes with MTX function in patients with RA. METHODS: We computed MTX dose per patient per year for the years 1988 to 1999 and plotted these against time, comparing the overall mean MTX dose before and after 1997, when dietary fortification with folic acid was instituted in the USA. Thirty-six subjects met eligibility criteria. RESULTS: Mean annual MTX dose was stable between 1988 and 1996 (12.4 +/- 4.0mg), but then rose linearly from 1997 to 1999 (16.6 +/- 5.1 mg, p < 0.001). CONCLUSIONS: This preliminary study suggests that folic acid supplementation may contribute to higher MTX dosing in patients with RA. PMID- 17914134 TI - Effects of various forms of calcium on body weight and bone turnover markers in women participating in a weight loss program. AB - OBJECTIVE: This study examined the effects of calcium intake on body weight, body fat, and markers of bone turnover in pre-menopausal adult women undergoing a 12 week weight loss program of diet and exercise. METHODS: Subjects were prescribed a 12 week diet with a 500 Kcal restriction containing about 750 mg calcium/day, exercised 3 times/week, and were given either placebo capsules, capsules of calcium lactate or calcium phosphate (daily dose about 800 mg calcium), or low fat milk (daily dose about 800 mg calcium). Subjects completed and returned daily diet diaries weekly. RESULTS: Daily calcium intake in mg from diet records + supplement assignment was: 788 +/- 175 (placebo), 1698 +/- 210 (Ca lactate), 1566 +/- 250 (Ca phosphate), 1514 +/- 225 (milk)(no significant differences among the calcium and milk groups). Each group had statistically significant changes in body weight (p < 0.01), but there were no significant differences among groups for the weight loss: 5.8 +/- 0.8 kg (placebo), 4.1 +/- 0.7 kg (Ca lactate), 5.4 +/- 1.3 kg (Ca phosphate), 4.2 +/- 0.8 kg (milk). Body fat was changed significantly in each group (p < 0.01), with milk group showing a little less change than the other groups. Serum bone specific alkaline phophatase activity, a bone synthesis marker, increased similarly in all groups (p < 0.001 within groups, no significance for changes among groups). In contrast, the Ca lactate group, but not other groups, had a drop in urine values for alpha helical peptide, a bone resorption marker (p < 0.05). CONCLUSION: For the conditions of this study, increased calcium intake, by supplement or milk, did not enhance loss of body weight or fat, though calcium lactate supplementation lowered values for a marker of bone degradation. PMID- 17914135 TI - Preliminary findings: 25(OH)D levels and PTH are indicators of rapid bone accrual in pubertal children. AB - OBJECTIVE: The objective of this study was to evaluate the role of serum levels of 25(OH)D and PTH on the accumulation of whole body bone mass in a cohort of children. METHODS: This was a longitudinal study (1.98 +/- 0.07 y) of sixty-nine children (89% Caucasian, 44% male) enrolled in a calcium supplementation trial. Bone area, bone mineral content (BMC) and density (BMD) of the whole body and radius were assessed using a QDR 2000 (Hologic, Inc) dual energy x-ray absorptiometer. Serum PTH and 25(OH)D were measured using radioimmunoassays. RESULTS: Vitamin D stores were inversely related gain in bone area (p < 0.002), BMC (p < 0.002) BMD (p < 0.027), as well as to PTH levels (p < 0.0001). Compared to those with adequate vitamin D stores (>34 ng/ml), those who had consistently low vitamin D stores (18 ng/ml) had a 8% larger gain in bone area (p < 0.05); 11% in BMC (p < 0.05) and no differences in gain in BMD; after adjusting for baseline bone measurements, race, gender, season measured, Tanner stage, and calcium intake. CONCLUSIONS: High normal PTH with low-normal 25(OH)D stores and moderate to high calcium intake may be beneficial to accruing larger bone size and BMC during puberty. PMID- 17914136 TI - The effect of consuming instant black tea on postprandial plasma glucose and insulin concentrations in healthy humans. AB - OBJECTIVE: To determine the effects of black tea on postprandial plasma glucose and insulin concentrations in healthy humans in response to an oral glucose load. METHODS: A four-way randomised, crossover trial was designed in which 16 healthy fasted subjects would consume 75g of glucose in either 250ml of water (control), 250ml of water plus 0.052g of caffeine (positive control) or 250 ml of water plus 1.0g or 3.0g of instant black tea. Blood samples were collected at fasting and at 30min intervals for 150min from commencement of drink ingestion. Glucose and insulin concentrations were measured using standard methodology. The tea was chemically characterised using colorimetric and HPLC methods. RESULTS: Chemical analysis showed that the tea was rich in polyphenolic compounds (total, 350mg/g). Results from only 3 treatment arms are reported because the 3.0g tea drink caused gastrointestinal symptoms. Plasma glucose concentrations <60min in response to the drinks were similar, but were significantly reduced at 120min (P<0.01), following ingestion of the 1.0g tea drink, relative to the control and caffeine drinks. Tea consumption resulted in elevated insulin concentrations compared with the control and caffeine drinks at 90min (P<0.01) and compared with caffeine drink alone at 150min (P<0.01). CONCLUSIONS: The 1.0g tea drink reduced the late phase plasma glucose response in healthy humans with a corresponding increase in insulin. This may indicate that the attenuation in postprandial glycemia was achieved as a result of an elevated insulin response following stimulation of pancreatic beta-cells. This effect may be attributable to the presence of phenolic compounds in the tea. PMID- 17914137 TI - Carpal tunnel syndrome. PMID- 17914138 TI - Evaluation of a Hong Kong Chinese version of a self-administered questionnaire for assessing symptom severity and functional status of carpal tunnel syndrome: cross-cultural adaptation and reliability. AB - OBJECTIVES: To evaluate the application of a translated version of an established self-administered questionnaire for carpal tunnel syndrome on Chinese patients in Hong Kong. DESIGN: Evaluation of an instrument tool. SETTING: Department of Orthopaedics and Traumatology, Queen Mary Hospital, Hong Kong; Holistic Medical Centre, Aberdeen, Hong Kong. PARTICIPANTS: Patients with carpal tunnel syndrome, translators. MAIN OUTCOME MEASURES: The adaptation was based on forward-backward translation from English to Chinese (Hong Kong) and vice versa. Meetings with translators, investigators, and patients were organised to generate an acceptable version of the questionnaire. A pilot study was carried out on 20 patients and subsequently minor adjustments were added. Fifty patients were recruited to validate the reliability and internal consistency of the questionnaire. RESULTS: The ordinality of response agreed with the original instrument. Test-retest reproducibility showed no significant difference between tests. The Pearson correlation coefficient ranged from 0.83 to 0.93. Internal consistency was good, at 0.85. CONCLUSION: Through the validation of the Hong Kong Chinese version of the questionnaire, we are able to produce an assessment tool for the local patients. Furthermore, we are able to create a platform for: (i) a cross-national and cross-cultural epidemiological comparison as well as a means of (ii) evaluating different types of treatments. PMID- 17914139 TI - Clinical experience of open carpal tunnel release and Camitz operation in elderly Chinese patients. AB - OBJECTIVE: To report preliminary experience on the Camitz operation for elderly Chinese patients in a Hong Kong public hospital. DESIGN: Retrospective study. SETTING: Tertiary referral hospital with hand surgery service in Hong Kong. PATIENTS: Between January 2000 and January 2004, patients with carpal tunnel syndrome having the Camitz operation were recruited. They were assessed using the measurements of pinch and grip power, sensation, the Kapandji score, and functional grading as well as complications encountered during the subsequent follow-up. INTERVENTION: The Camitz operation. RESULTS: A total of 21 patients (8 male and 13 female; mean age, 70 years) were recruited. The mean duration of follow-up was 15 months. There was significant improvement in pinch power, grip power, and hand functions, as well as a positive correlation between the functional score and the Kapandji score. No major complication was recorded. One patient with pre-existing osteoarthritis of the ring finger developed contracture of the proximal interphalangeal joint. CONCLUSION: The Camitz operation is a simple, effective, and safe means of improving hand function in elderly Chinese patients with long-standing carpal tunnel syndrome and thenar muscle atrophy. Newly acquired strength in antepulsion of thumb resulted in improved pinch, grip, and hand function in this patient group. PMID- 17914140 TI - Is regular follow-up scan for giant liver haemangioma necessary? AB - OBJECTIVES: To review the reliability of radiological diagnosis and need of regular scans for giant liver haemangioma, in terms of long-term outcome and management options. DESIGN: Retrospective study. SETTING: Division of Hepato biliary and Pancreatic Surgery, Prince of Wales Hospital, Hong Kong. PATIENTS: Patients with giant liver haemangioma noted on initial imaging from February 1996 to July 2006. MAIN OUTCOME MEASURES: Patient demographics, clinical assessments, management, and outcomes. RESULTS: There were 42 female and 22 male patients with a median age of 49 (range, 27-84) years with a suspected haemangioma. The median maximal diameter of the lesions was 5.5 cm (range, 4.0-20.3 cm). They were first detected by ultrasonography (n=45), contrast-enhanced computed tomographic scan (n=18), or magnetic resonance imaging (n=1). Besides regular follow-up scans, 22 patients were investigated further to confirm the diagnosis/exclude malignancy. Finally, 63 patients had a haemangioma and one had a hepatocellular carcinoma. Regarding the patients with haemangiomas, two were operated on for relief of pain and the rest were managed conservatively. The median duration of follow-up was 34 months. Most (54%) of the patients were asymptomatic, but in 17% the haemangioma enlarged to exceed its original size by more than 20%. There were no haemangioma associated complications. CONCLUSIONS: Majority of patients having giant liver haemangioma are asymptomatic and do not suffer complications. If the diagnosis is uncertain, selective further investigations may be necessary. Lesions with a confirmed diagnosis tend to remain static in size; performing regular scans for asymptomatic giant liver haemangiomas may not be necessary. PMID- 17914141 TI - Analogues of erectile dysfunction drugs: an under-recognised threat. AB - OBJECTIVES: To investigate the problem of drug analogue adulteration in male erectile dysfunction health products. DESIGN: Survey of over-the-counter male erectile dysfunction health products available in convenience stores and pharmacies in Hong Kong. SETTING: Tertiary referral centre for clinical toxicology analysis in Hong Kong. MAIN OUTCOME MEASURES: The pattern and extent of adulteration of male erectile dysfunction health products with sildenafil, tadalafil, and vardenafil as well as their structurally modified analogues. RESULTS: Of 26 products studied, one (4%) was found to contain undeclared sildenafil, while 14 (54%) contained drug analogues of different kinds. The latter included acetildenafil, hydroxyacetildenafil, hydroxyhomosildenafil, and piperidenafil. The first three were analogues of sildenafil and the last was an analogue of vardenafil. One young patient presented with ataxia after taking an acetildenafil-containing product. CONCLUSIONS: The positive rate of concealed drug analogues in male erectile dysfunction health products is alarmingly high. Such analogues are difficult to detect by ordinary laboratory methods, and might be used in an attempt to evade regulatory inspection. Without going through the stringent drug testing process, the adverse effects of these chemicals remain largely unknown and unpredictable. Effective surveillance system and control measures are needed urgently. The medical profession and the public should be alerted to this under-recognised threat. PMID- 17914142 TI - Perceived child behaviour problems, parenting stress, and marital satisfaction: comparison of new arrival and local parents of preschool children in Hong Kong. AB - OBJECTIVE: To compare parental perception of child behaviour problems, parenting stress, and marital satisfaction in new arrival and local parents. DESIGN: Cross sectional survey; semi-structured interview. SETTING: Maternal and Child Health Centres, social service centres, preschools. PARTICIPANTS: Parents of preschool children, including new arrival parents and local parents. MAIN OUTCOME MEASURES: Child behaviour problems, parenting stress, and marital satisfaction. RESULTS: After controlling for socio-economic factors, new arrival parents were more troubled by their children's behaviour problems and their parent-child interactions were more dysfunctional than those of local parents. There were no differences in parent-reported severity of child behaviour problems, parental distress, and marital satisfaction. New arrival parents reported difficulties in adapting to the new living environment and lack of social support. CONCLUSIONS: New arrival parents were more troubled by their children's behaviour, and their parent-child interactions were more dysfunctional than those of local parents. These might in part be related to their settlement difficulties. Parenting programmes should address their specific settlement needs. PMID- 17914143 TI - Clinical experience with a chronic pain management programme in Hong Kong Chinese patients. AB - OBJECTIVE: To describe experience with a chronic pain management programme in Hong Kong Chinese patients. DESIGN: Prospective study. SETTING: Regional hospital, Hong Kong. PARTICIPANTS: Patients with chronic pain who participated in the first six Comprehensive Out-patient Pain Engagement programmes between 2002 and 2005. INTERVENTION: Comprehensive Out-patient Pain Engagement is a 14-day structured, multidisciplinary out-patient programme conducted over 6 weeks. It includes pain education, cognitive re-conceptualisation, training in communication skills and coping strategies, graded physical exercises and functional activities training. It aims to improve patient function and quality of life, despite persistent pain. MAIN OUTCOME MEASURES: Changes in scores from baseline values after joining the programme, with respect to several assessment tools. These included the following: visual analogue pain scale, Pain Catastrophizing Scale, Patient Self-efficacy Questionnaire, Canadian Occupational Performance Measure, Medical Outcome Survey-Short Form 36 Questionnaire, and duration of physical tolerances, medication utilisation, and work status records. RESULTS: Forty-five patients were available for analysis. After the Comprehensive Out-patient Pain Engagement programme, improvements in Medical Outcome Survey Short Form 36 Questionnaire (role physical and vitality), Pain Catastrophizing Scale, Patient Self-efficacy Questionnaire, and Canadian Occupational Performance Measure were demonstrated (P<0.05). The duration of standing and sitting tolerances increased (P<0.05). An improvement in employment rate was also evident (P=0.01). CONCLUSION: The initial results of our management programme in Chinese patients with chronic pain are encouraging. This type of programme should be promoted more widely in this group of patients, as it appears to improve physical function, psychological well-being, and productivity. PMID- 17914144 TI - Seasonal variations in non-traumatic major lower limb amputation in Hong Kong Chinese diabetic patients. AB - OBJECTIVES: To study the relationship between hospitalisation for diabetic foot complications, non-traumatic major lower limb amputations, and seasonal variation in humidity and temperature. DESIGN: Retrospective study. SETTING: Regional hospital, Hong Kong. PATIENTS: Diabetic patients hospitalised for lower limbs infection during the inclusive period 1995 to 2004. MAIN OUTCOME MEASURES: Monthly incidence of admissions for diabetic foot sepsis and non-traumatic non neoplastic major lower limb amputations correlated with the monthly average humidity and temperature. RESULTS: During the study period, a total of 770 patients contributed to 1285 episodes of hospitalisation and ensued 208 corresponding major lower limb amputations. Poisson regression study showed that the monthly incidences of hospitalisation as well as amputations were related to the monthly average temperature (P<0.001 and =0.0012, respectively) but not the monthly average humidity (P=0.1560 and 0.6332, respectively). CONCLUSION: The warm and humid weather of Hong Kong exerts a seasonal variation on the diabetic foot infection presentations. Warm temperature aggravates the severity of infection and precipitates amputation. More intensive patient education and clustering of medical services in late winter and spring might reduce the incidence of diabetic foot amputation, which is a preventable complication. PMID- 17914145 TI - Maggot debridement therapy in chronic wound care. AB - OBJECTIVE: To review the current evidence on the mechanism of actions and clinical applications of maggot debridement therapy. DATA SOURCES: Literature search of PubMed and Medline was performed up to January 2007. STUDY SELECTION: Original and major review articles related to maggot debridement therapy were reviewed. Key words used in the literature search were 'maggot debridement therapy', 'wound healing', and 'chronic wound management'. DATA EXTRACTION: All relevant English and Chinese articles. DATA SYNTHESIS: The mechanism of such maggot therapy was shown to be due to the debridement, disinfection, and wound healing enhancement actions of maggot excretions/secretions. The efficacy of maggot debridement therapy in chronic wound management has been demonstrated in chronic venous ulcers, pressure ulcers, and diabetic ulcers. There is also a new delivery system for the excretions/secretions, which has been shown to be as effective as using live maggots. CONCLUSIONS: Maggot debridement therapy has been shown to be a safe and effective means of chronic wound management. However, there are a number of limitations when considering its local applicability. Future development of the delivery system may help to overcome some of these limitations and improve its acceptability. PMID- 17914146 TI - Obstetric cholestasis in Hong Kong--local experience with eight consecutive cases. AB - Obstetric cholestasis is associated with maternal morbidity and adverse foetal outcomes. No information on local incidence is available. We present our experience with eight consecutive cases of obstetric cholestasis diagnosed between January 2003 and December 2005 in a regional hospital in Hong Kong. Three patients presented with pruritus without rash, three with impaired liver function, and two with elevated blood pressure postpartum. Meconium-stained liquor was present in five patients and four had spontaneous preterm delivery (between 34 and 36 weeks). The higher the bile acid level, the more marked the prematurity (correlation coefficient, -0.771; P=0.025). All those presenting with itchiness delivered preterm. Two patients developed pre-eclampsia. The rates of labour induction and abdominal delivery were both 38%. Heightened awareness among clinicians is required to recognise patients with obstetric cholestasis. Affected pregnancies are associated with meconium passage and prematurity. In our locality, affected women may also have an increased risk of pre-eclampsia. In affected women, the bile acid level is useful in assessing the risk of prematurity. PMID- 17914147 TI - Severe coagulopathy associated with white-lipped green pit viper bite. AB - The authors report a case of Trimeresurus albolabris (white-lipped green pit viper) bite in a 6-year-old girl living in rural Yuen Long. Despite repeated use of Agkistrodon halys antivenin, the patient developed severe coagulopathy with defibrination syndrome on the fourth day of envenomation, which was also refractory to therapy with fresh frozen plasma. When treatment was switched to green pit viper antivenin, the coagulopathy resolved promptly. The case is illustrative of the potential lethality to children of snakebites in Hong Kong and suggests that the A halys antivenin may not be effective for the treatment of T albolabris bites. PMID- 17914148 TI - Spontaneous rupture of the left renal collecting system during pregnancy. AB - Spontaneous rupture of a normal renal collecting system during pregnancy is uncommon and all reported cases have occurred in right kidneys. We report a case of spontaneous rupture of the left renal collecting system during pregnancy. A 33 year-old pregnant woman presented with left loin and lower abdominal pain, and signs of preterm labour, at 32 weeks' gestation. An emergency caesarean section was performed for foetal distress but the left loin pain did not subside after delivery. Ultrasonography and a computed tomogram showed a left perinephric collection and urine extravasation, compatible with rupture of the renal collecting system. A percutaneous nephrostomy was inserted and the symptoms subsided. A summary of the literature discussing management of this clinical situation is provided. PMID- 17914149 TI - Bryan artificial cervical disc arthroplasty in a patient with Klippel-Feil syndrome. AB - Technological advances have made more options available for surgical intervention in spinal disorders. From spinal fusion to artificial disc implantation, these advancements have brought great benefits, allowing preservation of spinal motion and flexibility after intervertebral discectomy. Yet the use of artificial discs as a treatment for congenital spinal disorders has been documented in only a handful of publications. We report a case where a Bryan artificial cervical disc arthroplasty was used to maintain and preserve the mobility and function of the cervical motion segments adjacent to fused vertebral lesions in a 33-year-old woman with Klippel-Feil syndrome who presented with chronic neck pain and signs of early myelopathy. The rationales for using the Bryan disc prosthesis system in patients with Klippel-Feil syndrome and its advantages over conventional surgical interventions are discussed. PMID- 17914150 TI - Arthroscopy-assisted combined external and internal fixation of a pilon fracture of the tibia. AB - There are serious problems with existing methods of treating pilon fractures of the tibia caused by high-energy trauma. The method chosen to treat these fractures should not raise the risk of infection while effectively restoring the joint surface. We successfully treated a 42-year-old male patient with a pilon fracture caused by high-energy trauma using an arthroscopy-assisted unilateral external fixator and minimally invasive internal osteosynthesis. We used arthroscopy to reposition the fracture fragments and restore the joint surface. The fracture fragments were fixed with screws immediately after being repositioned. We believe that arthroscopy-assisted combined external and minimally invasive internal fixation is the treatment of choice for these fractures. We used external fixation to improve the fracture alignment, arthroscopy for restoring the joint surface, and minimally invasive screws to ensure fragment stability. PMID- 17914151 TI - Carcinoid tumour of the kidney in a Chinese woman presenting with loin pain. AB - Renal carcinoid tumours are uncommon. The aetiology is not yet fully understood and there is still no useful diagnostic tool for detecting them. We report our experience managing a Chinese woman with a primary renal carcinoid tumour. PMID- 17914152 TI - Worsening back pain in pregnancy. PMID- 17914153 TI - Anterior myocardial infarction due to left anterior descending artery occlusion with stand-alone ST elevation in leads I and aVL. PMID- 17914154 TI - A mini review of P values from Hong Kong Medical Journal. PMID- 17914156 TI - Signaling functions of free radicals superoxide & nitric oxide under physiological & pathological conditions. AB - Superoxide and nitric oxide are ubiquitous physiological free radicals that are responsible for many pathological disorders. Both radicals by themselves are relatively harmless but are the precursors of many toxic species such as peroxy and hydroxyl radicals, hydrogen peroxide, and peroxynitrite. However, it has been shown now that both superoxide and nitric oxide are also able to perform important signaling functions in physiological and pathophysiological processes. Wrongly named "superoxide," the radical anion of dioxygen is not a super-oxidant but the strong super-nucleophile, an efficient catalyst of heterogenic nucleophilic reaction. Due to this, superoxide plays an important role in many enzymatic processes such as the phosphorylation and activation of numerous protein kinases. On the other hand, superoxide inhibits the activation of phosphatases, the enzymes catalyzed by dephosphorylation of protein kinases. We suggest that superoxide catalyzes these enzymatic processes as a result of its nucleophilic properties. Another important physiological function of superoxide and nitric oxide is their competition for the interaction with mitochondrial cytochrome c oxidase. Disturbance of superoxide/nitric oxide balance leads to the dysfunction of mitochondria and the enhancement of apoptosis and oxidative stress, which are primary causes of various pathological disorders and aging. In conclusion, interplay between superoxide and nitric oxide, one of major factors of aging development, is considered. PMID- 17914157 TI - The important role of lipid peroxidation processes in aging and age dependent diseases. AB - Any change in the cell membrane structure activates lipoxygenases (LOX). LOX transform polyunsaturated fatty acids (PUFAs) to lipidhydroperoxide molecules (LOOHs). When cells are severely wounded, this physiological process switches to a non-enzymatic lipid peroxidation (LPO) process producing LOO* radicals. These oxidize nearly all-biological molecules such as lipids, sugars, and proteins. The LOO* induced degradations proceed by transfer of the radicals from cell to cell like an infection. The chemical reactions induced by LO* and LOO* radicals seem to be responsible for aging and induction of age dependent diseases.Alternatively, LO* and LOO* radicals are generated by frying of fats and involve cholesterol-PUFA esters and thus induce atherogenesis. Plants and algae are exposed to LOO* radicals generating radiation. In order to remove LOO* radicals, plants and algae transform PUFAs to furan fatty acids, which are incorporated after consumption of vegetables into mammalian tissues where they act as excellent scavengers of LOO* and LO* radicals. PMID- 17914158 TI - ASK family proteins in stress response and disease. AB - Cells are continuously exposed to reactive oxygen species (ROS) generated by aerobic metabolism. Excessively generated ROS causes severe dysfunctions to cells as oxidative stress. On the other hand, there is increasing evidence that ROS plays important roles as a signaling intermediate that induces a wide variety of cellular responses such as proliferation, differentiation, senescence, and apoptosis. To transmit physiological ROS-mediated signals and to adapt to oxidative stress, cells are equipped with various intracellular signal transduction systems, represented by mitogen-activated protein kinase (MAPK) cascades. Apoptosis signal-regulating kinase 1 (ASK1) is an upstream regulator of the stress-activated MAPK cascades and has been shown to play critical roles in ROS-mediated cellular responses. Here, we highlight the roles of members of the ASK family, which consists of ASK1 and newly characterized ASK2, in ROS signaling with their possible involvement in human diseases. PMID- 17914159 TI - Trans fatty acids in membranes: the free radical path. AB - The double bond geometry of most of the naturally occurring unsaturated fatty acid residues is cis. Due to the relevance of fatty acids as structural components of cell membranes and as biologically active molecules, the change of the cis geometry means a change of the associated functions and activities. The finding that the cis to trans isomerization is effective in phospholipids by the intervention of radical species led to the discovery that there can indeed occur an endogenous formation of trans fatty acids, whose significance in biological systems started to be addressed with in vitro and in vivo studies. Studies of liposome models simulating the formation of isomerizing species and evaluating their ability to interact with the hydrophobic part of the membrane bilayer has contributed to the gain in knowledge of the fundamental features of the lipid isomerization in membranes. Further work is in progress for the identification of the real culprits of the in vivo lipid isomerization, and recent results are shown on oleic acid micelles, where *NO2 radicals are not able to induce double bond isomerization in comparison with amphiphilic thiol, such as 2 mercaptoethanol. H2S and sulfur-containing amino acid residues are two of the possible species involved in this process at a biological level. An update of the scenario of the geometrical isomerization in membranes by free radicals is provided, together with applications and perspectives in life sciences. PMID- 17914160 TI - Carotenoids and flavonoids contribute to nutritional protection against skin damage from sunlight. AB - The concept of photoprotection by dietary means is gaining momentum. Plant constituents such as carotenoids and flavonoids are involved in protection against excess light in plants and contribute to the prevention of UV damage in humans. As micronutrients, they are ingested with the diet and are distributed into light-exposed tissues, such as skin or the eye where they provide systemic photoprotection. beta-Carotene and lycopene prevent UV-induced erythema formation. Likewise, dietary flavanols exhibit photoprotection. After about 10-12 weeks of dietary intervention, a decrease in the sensitivity toward UV-induced erythema was observed in volunteers. Dietary micronutrients may contribute to life-long protection against harmful UV radiation. PMID- 17914161 TI - Bioenergetic and antioxidant properties of coenzyme Q10: recent developments. AB - For a number of years, coenzyme Q (CoQ10 in humans) was known for its key role in mitochondrial bioenergetics; later studies demonstrated its presence in other subcellular fractions and in plasma, and extensively investigated its antioxidant role. These two functions constitute the basis on which research supporting the clinical use of CoQ10 is founded. Also at the inner mitochondrial membrane level, coenzyme Q is recognized as an obligatory co-factor for the function of uncoupling proteins and a modulator of the transition pore. Furthermore, recent data reveal that CoQ10 affects expression of genes involved in human cell signalling, metabolism, and transport and some of the effects of exogenously administered CoQ10 may be due to this property. Coenzyme Q is the only lipid soluble antioxidant synthesized endogenously. In its reduced form, CoQH2, ubiquinol, inhibits protein and DNA oxidation but it is the effect on lipid peroxidation that has been most deeply studied. Ubiquinol inhibits the peroxidation of cell membrane lipids and also that of lipoprotein lipids present in the circulation. Dietary supplementation with CoQ10 results in increased levels of ubiquinol-10 within circulating lipoproteins and increased resistance of human low-density lipoproteins to the initiation of lipid peroxidation. Moreover, CoQ10 has a direct anti-atherogenic effect, which has been demonstrated in apolipoprotein E-deficient mice fed with a high-fat diet. In this model, supplementation with CoQ10 at pharmacological doses was capable of decreasing the absolute concentration of lipid hydroperoxides in atherosclerotic lesions and of minimizing the size of atherosclerotic lesions in the whole aorta. Whether these protective effects are only due to the antioxidant properties of coenzyme Q remains to be established; recent data point out that CoQ10 could have a direct effect on endothelial function. In patients with stable moderate CHF, oral CoQ10 supplementation was shown to ameliorate cardiac contractility and endothelial dysfunction. Recent data from our laboratory showed a strong correlation between endothelium bound extra cellular SOD (ecSOD) and flow-dependent endothelial mediated dilation, a functional parameter commonly used as a biomarker of vascular function. The study also highlighted that supplementation with CoQ10 that significantly affects endothelium-bound ecSOD activity. Furthermore, we showed a significant correlation between increase in endothelial bound ecSOD activity and improvement in FMD after CoQ10 supplementation. The effect was more pronounced in patients with low basal values of ecSOD. Finally, we summarize the findings, also from our laboratory, on the implications of CoQ10 in seminal fluid integrity and sperm cell motility. PMID- 17914162 TI - The rational design of beta cell cytoprotective gene transfer strategies: targeting deleterious iNOS expression. AB - Islet transplantation represents a promising therapeutic strategy for the treatment of type 1 diabetes mellitus (T1DM) [Hakim and Papalois (Ann Ital Chir 75:1-7, 2004); Jaeckel et al. (Internist (Berl) 45:1268-1280, 2004); Sutherland et al. (Transplant Proc 36:1697-1699, 2004)]. The insulin-secreting pancreatic beta cells of the islet allograft are, however, subject to recurrent immune mediated damage. Principal among the molecular culprits involved in this destructive process is the proinflammatory cytokine IL-1beta. IL-1beta-induced beta cell destruction may be mediated by the generation of NO and/or ROS, although the relative importance of NO and ROS in this process remains unclear. This study broadly encompassed three arms of investigation: the first of these was geared toward the establishment of a robust in vitro cell system for the study of IL-1beta-induced pathophysiology; the second arm aimed to provide a comparative analysis of the gene transfer profiles of the three most commonly used gene transfer vehicles, namely plasmid vectors, adenoviral vectors, and lentiviral vectors, in the aforementioned cell system; the final arm aimed to screen an array of potentially cytoprotective gene transfer strategies incorporating the optimal gene transfer vectors. Briefly, we established an in vitro beta cell system that accurately reflected primary beta cell cytokine induced pathophysiology. That is, IL-1beta exposure (100 U/ml) induced a time dependent decrease in rat insulinoma (RIN) cell viability, which coincided with an induction in iNOS expression and nitrite accumulation. Gene transfer studies using plasmid, adenoviral, or lentiviral vectors underscored the superiority of viral vector-based gene transfer strategies for the manipulation of this beta cell line. Using these vectors, we provide evidence that NF-kappaB-based iNOS inhibition confers significant protection against IL-1beta-induced damage whereas antioxidant overexpression fails to provide protection. Conferred cytoprotection was associated with a suppression of iNOS expression and nitrite accumulation. From a therapeutic standpoint, gene transfer strategies employing efficient viral vectors to target iNOS activation may harbour therapeutic potential in preserving beta cell survival against proinflammatory cytokine exposure. PMID- 17914163 TI - Photochemical properties of camptothecin in the presence of copper(II) ions: the role of radicals as prospective species in photodynamic therapy. AB - Camptothecin (CPT) is an anticancer drug that inhibits topoisomerase I (Topo I) by forming a ternary DNA-CPT-Topo I complex. However, it has also been shown that UVA-irradiated CPT in the absence of Topo I produces significant DNA damage to cancer cells. In this work, we explored and identified free radicals generated in these processes. From the low-temperature EPR spectrum of Cu(II)-CPT complex, a proximity between Cu(II) ion and 20-hydroxy group of lactone E ring of CPT is proposed. Upon irradiation (lambda = 365 nm) of the Cu(II)-CPT complex in de oxygenated dimethylsulfoxide (DMSO), the EPR signal of Cu(II) measured in situ at room temperature shows formal first-order exponential decay with a formal half life of 11 min. By the use of a specific Cu(I) chelating agent, neocuproine, it was shown that, during this process, Cu(II) is reduced to Cu(I). The loss in EPR signal intensity of the Cu(II)-CPT complex upon irradiation is accompanied by the appearance of a new EPR signal at g approximately 2.0022. Application of the spin trap nitrosodurene (ND) revealed that the main radical product formed upon continuous irradiation of CPT in DMSO solutions is the hydroxyl radical (trapped in DMSO as the *CH3 adduct) and superoxide radical. Application of 2,2,6,6 tetramethyl-4-piperidinol has revealed that irradiation of CPT in aerated DMSO solution also leads to formation of singlet oxygen (1O2). Our spectroscopic experiments indicate that CPT is a promising photosensitizer and that radicals and singlet oxygen generated upon illumination play a central role in DNA cleavage and in the induction of apoptosis in cancer cells. PMID- 17914164 TI - Anti-oxidants from green tea and pomegranate for chemoprevention of prostate cancer. AB - Among males, prostate cancer has become the second leading cause of cancer related deaths in North America, with similar trends in many Western and developing countries. One way to control prostate cancer is through chemoprevention, which refers to the administration of synthetic or naturally occurring agents to block, reverse, or delay the process of carcinogenesis. For a variety of reasons, the most important of which is human acceptance, for chemopreventive intervention, naturally occurring diet-based agents are preferred. Prostate cancer is an ideal candidate disease for chemopreventive intervention, because it grows very slowly, likely for decades, before symptoms arise and a diagnosis is finally established, it has a long latency period, and it is typically diagnosed in men >50 years of age. Most chemopreventive agents are antioxidant in nature. We have been defining the usefulness of dietary anti oxidants for chemoprevention of prostate and other cancers. It is increasingly appreciated that some of these dietary anti-oxidants are nature's gift molecules endowed with cancer preventive and therapeutic properties. This review will focus on prostate cancer chemopreventive effects of polyphenolic anti-oxidants derived from green tea and pomegranate. It is a challenge to custom-tailor these gift molecules as cocktails in concentrations that can easily be consumed by humans for delaying prostate and other cancers. PMID- 17914165 TI - Alterations in antioxidant enzymes during aging in humans. AB - The oxidative stress theory of aging offers the best mechanistic elucidation of the aging phenomenon and other age-related diseases. The susceptibility of an individual depends on the antioxidant status of the body. In humans, the antioxidant system includes a number of antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT), nonenzymatic antioxidants such as glutathione (GSH), protein -SH, ascorbic acid, and uric acid, and dietary antioxidants. Antioxidant enzymes form the first line of defense against reactive oxygen species. In an earlier report, we showed that the plasma antioxidant potential in humans decreases as a function of age and that there are compensatory mechanisms operating in the body which are induced to maintain the antioxidant capacity during aging. In the present study, we report the relationship between human aging and antioxidant enzymes SOD and CAT; we also correlate the activity of these enzymes with the antioxidant capacity of the plasma. Our results show a significantly higher plasma SOD and CAT activity in older individuals than in younger individuals. The induction in activity of SOD and CAT during human aging may be a compensatory response of the individual to an increased oxidative stress. PMID- 17914166 TI - Role of ascorbic acid in scavenging free radicals and lead toxicity from biosystems. AB - Free radicals are reactive species that are responsible for damaging normal cells and creating diseases in humans. Antioxidants from natural resources or as supplements can scavenge these radicals. A MedLine search indicates that vitamin C is the most investigated antioxidant responsible for the elimination of free radicals. Its chelating property for the removal of neurotoxic lead, which creates oxidative stress in the human biosystem, was investigated and results indicate its great potential as a lead-detoxifying agent. PMID- 17914167 TI - Carbofuran-induced oxidative stress in mammalian brain. AB - Chronic exposure to carbofuran, a carbamate pesticide, via oral administration has been reported to generate reactive oxygen species (ROS) in rat brain. However, information regarding the effect of short-term intraperitoneal (i.p.) carbofuran intoxication on oxidative stress is lacking. In the present study, the effect of carbofuran on oxidative indices in brain of Wistar rats has been determined by exposing the animals to three subacute concentrations (0.2, 0.4 and 0.8 mg/kg body weight) equivalent to 10, 20, and 40%, respectively, of its LD50 (i.p.) for 24 h. Rat liver has been used as a positive control. The results demonstrated that carbofuran treatment at the 3 concentrations tested caused significant increase in lipid peroxidation (LPO) by 12.50, 34.38, and 59.38%, respectively. The increased oxidative stress at same pesticide concentrations significantly induced activities of antioxidant enzymes such as superoxide dismutase (SOD) and catalase in rat brain; the impact on catalase being more marked only at high-pesticide doses (0.4 and 0.8 mg/kg body weight). Carbofuran also caused reduction in protein content of rat tissues tested. Rat brain was more severely affected by carbofuran than liver. The results clearly demonstrated that i.p. administration of carbofuran accelerated oxidative stress in rat brain in a dose-dependent manner. PMID- 17914168 TI - From nutraceuticals to pharmaceuticals to nanopharmaceuticals: a case study in angiogenesis modulation during oxidative stress. AB - This report reviews the potential applications of nanotechnology in various therapeutics and diagnostics areas with special emphasis on key frontiers in angiogenesis modulation using naturally driven drug targets including compounds that modulate oxidative stress and inflammatory pathways for the potential treatment of vascular, cancer, inflammatory, and ocular disorders. Recent advances of the nanotechnology mediated gene delivery are also described in this paper. PMID- 17914170 TI - A comparison of protein quantitation assays for biopharmaceutical applications. AB - Dye-based protein determination assays are widely used to estimate protein concentration, however various reports suggest that the response is dependent on the composition and sequence of the protein, limiting confidence in the resulting concentration estimates. In this study a diverse set of model proteins representing various sizes of protein and covalent modifications, some typical of biopharmaceuticals have been used to assess the utility of dye-based protein concentration assays. The protein concentration assays (Bicinchoninic acid (BCA), Bradford, 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA), DC, Fluorescamine and Quant-i) were compared to the 'gold standard' assay, quantitative amino acid analysis (AAA). The assays that displayed the lowest variability between proteins, BCA and DC, also generated improved estimates when BSA was used as a standard, when compared to AAA derived concentrations. Assays read out by absorbance tended to display enhanced robustness and repeatability, whereas the fluorescence based assays had wider quantitation ranges and lower limits of detection. Protein modification, in the form of glycosylation and PEGylation, and the addition of excipients, were found to affect the estimation of protein concentration for some of the assays when compared to the unmodified protein. We discuss the suitability and limitations of the selected assays for the estimation of protein concentration in biopharmaceutical applications. PMID- 17914171 TI - Production of 1,3-propanediol from glycerol by recombinant E. coli using incompatible plasmids system. AB - 1,3-Propanediol (1,3-PD) has numerous applications in polymers, cosmetics, foods, lubricants, and medicines as a bifunctional organic compound. The genes for the production of 1,3-PD in Klebsiella pneumoniae, dhaB, which encodes glycerol dehydratase, and dhaT, which encodes 1,3-PD oxidoreductase, and gdrAB, which encodes glycerol dehydratase reactivating factor, are naturally under the control of different promoters and are transcribed in different directions. These genes were coexpressed in E. coli using two incompatible plasmids (pET28a and pET22b) in the presence of selective pressure. The recombinant E. coli coexpressed the glycerol dehydratase, 1,3-propanediol oxidoreductase and reactivating factor for the glycerol dehydratase at high levels. In a fed-batch fermentation of glycerol and glucose, the recombinant E. coli containing these two incompatible plasmids consumed 14.3 g/l glycerol and produced 8.6 g/l 1,3-propanediol. In the substitution case of yqhD (encoding alcohol dehydrogenase from E. coli) for dhaT, the final 1,3-propanediol concentration of the recombinant E. coli could reach 13.2 g/l. PMID- 17914172 TI - Time-course determination of plasmid content in eukaryotic and prokaryotic cells using real-time PCR. AB - A Real-Time PCR method was developed to monitor the plasmid copy number (PCN) in Escherichia coli and Chinese hamster ovary (CHO) cells. E. coli was transformed with plasmids containing a ColE1 or p15A origin of replication and CHO cells were transfected with a ColE1 derived plasmid used in DNA vaccination and carrying the green fluorescent protein (GFP) reporter gene. The procedure requires neither specific cell lysis nor DNA purification and can be performed in <30 min with dynamic ranges covering 0.9 pg-55 ng, and 5.0 pg-2.5 ng of plasmid DNA (pDNA) for E. coli and CHO cells, respectively. Analysis of PCN in E. coli batch cultures revealed that the maximum copy number per cell is attained in mid-exponential phase and that this number decreases on average 80% towards the end of cultivation for both types of plasmids. The plasmid content of CHO cells determined 24 h post-transfection was around 3 x 104 copies per cell although only 37% of the cells expressed GFP one day after transfection. The half-life of pDNA was 20 h and around 100 copies/cell were still detected 6 days after transfection. PMID- 17914173 TI - Binding of MmeI restriction-modification enzyme to its specific recognition sequence is stimulated by S-adenosyl-L-methionine. AB - Restriction endonucleases serve as a very good model for studying specific protein-DNA interaction. MmeI is a very interesting restriction endonuclease, but although it is useful in Serial Analysis of Gene Expression, still very little is known about the mechanism of its interaction with DNA. MmeI is a unique enzyme as besides cleaving DNA it also methylates specific sequence. For endonucleolytic activity MmeI requires Mg(II) and S-adenosyl-l-methionine (AdoMet). AdoMet is a methyl donor in the methylation reaction, but its requirement for DNA cleavage remains unclear. In the present article we investigated MmeI interaction with DNA with the use of numerous methods. Our electrophoretic mobility shift assay revealed formation of two types of specific protein-DNA complexes. We speculate that faster migrating complex consists of one protein molecule and one DNA fragment whereas, slower migrating complex, which appears in the presence of AdoMet, may be a dimer or multimer form of MmeI interacting with specific DNA. Additionally, using spectrophotometric measurements we showed that in the presence of AdoMet, MmeI protein undergoes conformational changes. We think that such change in the enzyme structure, upon addition of AdoMet, may enhance its specific binding to DNA. In the absence of AdoMet MmeI binds DNA to the much lower extent. PMID- 17914174 TI - Monoclonal antibodies recognize conformational epitopes on wild-type and recombinant mutant amidases from pseudomonas aeruginosa. AB - Hybridoma technology was used to raise monoclonal antibodies (MAbs) against wild type amidase from Pseudomonas aeruginosa. Hybridoma clones secreting polyol responsive MAbs (PR-MAbs) were screened that bind antigen tightly. but release under mild- and non-denaturing elution conditions, which can be used as ligands in immunoaffinity chromatography. Two of these hybridoma clones (C9E4 and B1E4) secreting MAbs against wild-type amidase were selected in order to check if they are PR-MAbs by using ELISA-elution assay. These hybridoma cell lines secreted MAbs of IgG class which were purified in a single step by Protein A-Sepharose CL 4B chromatography, which revealed two protein bands on SDS-PAGE. Specificity studies of MAb C9E4 revealed that it recognized a common epitope on wild-type and mutant T103I amidases as determined by direct ELISA, as well as by Western blotting under native conditions. This MAb exhibited a higher-affinity constant (K) for the mutant T103I amidase than for the wild-type enzyme. However, this MAb did not recognize either wild-type or mutant T103I enzymes under denaturing conditions suggesting that it binds to a conformation-sensitive epitope on amidase molecule. On the other hand, it also does not recognize either native or denatured forms of mutant C91A amidase suggesting that this substitution disrupted the conformational epitope present on amidase molecule. Furthermore, MAb C9E4 inhibited about 80% of wild-type amidase activity, whereas it activated about 80% of mutant amidase (T103I) activity. However, this MAb did not affect mutant C91A amidase activity which is in agreement with other results presented in this work. The data presented in this work suggest that this MAb acts as a powerful probe to detect conformational changes in native and denatured amidases as well as to differentiate wild-type and mutant (T103I and C91A) amidases. PMID- 17914175 TI - Optimizing expression of Streptococcus pneumoniae surface protein a, PspA: serocross-reactivity within families of antisera induced against clades 1 and 3. AB - Streptococcus pneumoniae is the agent responsible for infections such as pneumonia, otitis media, and meningitis. Among virulence factors, the Pneumococcal surface protein A (PspA) has been shown to be immunogenic and protective in mice, and is thus a good vaccine candidate. PspA has been classified into 6 clades and 3 families. Initially, pspA fragments, clades 1 and 3, were cloned into the pAE-6His expression vector. Proteins were expressed in Escherichia coli BL21(DE3) and purified by affinity and anion exchange chromatographies, with a yield of 11 mg/l of culture. Due to plasmid instability in E. coli, another construct using pspA1 was obtained based on pET-37b(+), which was shown to be stable in E. coli and increased the yield approximately 3-fold. Our results show good conditions for scale-up. Sera from immunized mice recognized PspA in total extracts of S. pneumoniae strains: anti-rPspA1p sera recognized native PspA clades 1 (+++), 2 (++) and 4 (+) and anti-rPspA3p sera recognized PspA clades 1 (+), 2 (+), 3 (+++) and 4 (+). The cross-reactivity pattern obtained confirms the notion that proteins from both families should be included for development of a broad-coverage vaccine; lower-cross reactivity between rPspAs of family 2 indicates that it may be necessary to include 2 proteins from this family. PMID- 17914176 TI - Expression and molecular characterization of ZmMYB-IF35 and related R2R3-MYB transcription factors. AB - R2R3-MYB transcription factors play many important roles in higher plants including the regulation of secondary metabolism, the control of cell shape, and in the response to various stress conditions. In spite of their large number and significance, very few of these genes have been functionally characterized in monocots. Here, we describe the characterization of ZmMYB-IF35 from maize. Using GAL4 fusion constructs, we show that ZmMYB-IF35 possesses the ability to bind DNA in a sequence specific manner and activate transcription in yeast. We also show that ZmMYB-IF35 is capable of binding to the a1 promoter in planta, but it is not sufficient for activation of a1 transcription. Interestingly, a chimeric protein consisting of the MYB domain from ZmMYB-IF35 and the non-MYB C-terminal region of P1, a closely related R2R3-MYB protein, activated transcription from the a1 promoter in planta, suggesting that regions outside the conserved R2R3-MYB domain contribute to regulatory specificity. In situ hybridization experiments demonstrate that ZmMYB-IF35 expresses primarily in epidermal and vascular cells, while its rice ortholog, OsMYB-IF, displays a broad expression pattern in aerial parts of the plant. Together, our results provide novel insights on the participation of ZmMYB-IF35 and related genes in the regulation of secondary metabolic pathways in the grasses. PMID- 17914177 TI - Isolation and amplification of genomic DNA from recalcitrant dried berries of black pepper (Piper nigrum L.)--a medicinal spice. AB - Black pepper is an important medicinal spice traded internationally. The extraction of high quality genomic DNA for PCR amplification from dried black pepper is challenging because of the presence of the exceptionally large amount of oxidized polyphenolic compounds, polysaccharides and other secondary metabolites. Here we report a modified hexadecyl trimethyl ammonium bromide (CTAB) protocol by incorporating potassium acetate and a final PEG precipitation step to isolate PCR amplifiable genomic DNA from dried and powdered berries of black pepper. The protocol has trade implication as it will help in the PCR characterization of traded black peppers from different countries. PMID- 17914178 TI - History of plant tissue culture. AB - Plant tissue culture, or the aseptic culture of cells, tissues, organs, and their components under defined physical and chemical conditions in vitro, is an important tool in both basic and applied studies as well as in commercial application. It owes its origin to the ideas of the German scientist, Haberlandt, at the begining of the 20th century. The early studies led to root cultures, embryo cultures, and the first true callus/tissue cultures. The period between the 1940s and the 1960s was marked by the development of new techniques and the improvement of those that were already in use. It was the availability of these techniques that led to the application of tissue culture to five broad areas, namely, cell behavior (including cytology, nutrition, metabolism, morphogenesis, embryogenesis, and pathology), plant modification and improvement, pathogen-free plants and germplasm storage, clonal propagation, and product (mainly secondary metabolite) formation, starting in the mid-1960s. The 1990s saw continued expansion in the application of the in vitro technologies to an increasing number of plant species. Cell cultures have remained an important tool in the study of basic areas of plant biology and biochemistry and have assumed major significance in studies in molecular biology and agricultural biotechnology. The historical development of these in vitro technologies and their applications are the focus of this chapter. PMID- 17914179 TI - Generation and characterization of mice with Myh9 deficiency. AB - Mutant alleles of MYH9 encoding a class II non-muscle myosin heavy chain-A (NMMHC IIA) have been linked to hereditary megathrombocytopenia with or without additional clinical features that include sensorineural deafness, cataracts, and nephritis. To assess its biological role in the affected targets, particularly the inner ear, we have generated and characterized mice with Myh9 deficiency. These mice were generated using the XA136 ES cell line (BayGenomics, http://baygenomics.ucsf.edu/) carrying gene trap insertion in Myh9, within the intron flanking exons 4 and 5. Mice heterozygous for the Myh9 null allele, Myh9 +/- were expanded on C57BL/6J background. Intercross of the Myh9 +/- mice did not yield Myh9 -/- pups, indicating embryonic lethality, subsequently determined to occur at or before E7.5, thus precluding a post-natal analysis of the effects of complete Myh9 deficiency. The heterozygous mice were normal for their hearing, parameters of platelet integrity and renal function despite their Myh9 haplo insufficiency. In addition, the age-dependent auditory threshold of the Myh9 +/- mice and their wild type littermates, spanning from 3 to 12 months of age, were similar indicating that Myh9 haplo-insufficiency does not contribute towards accelerated age-related hearing loss (AHL). The embryonic lethality associated with the complete Myh9 deficiency establishes a critical role for this non-muscle myosin in fetal development. The results of these studies do not support the Myh9 haploinsufficiency as a pathogenic factor in the etiology of auditory dysfunction. PMID- 17914180 TI - Magnetic resonance microscopy and immunohistochemistry of the CNS of the mutant SOD murine model of ALS reveals widespread neural deficits. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that primarily affects motor neurons and descending motor tracts of the CNS. We have evaluated the CNS of a murine model of familial ALS based on the over-expression of mutant human superoxide dismutase (mSOD; G93A) using magnetic resonance microscopy (MRM) and immunohistochemistry (IHC). Three-dimensional volumetric analysis was performed from 3D T2*-weighted images acquired at 17.6 T at isotropic resolutions of 40 mum. Compared to controls, mSOD mice had significant reductions in the volumes of total brain, substantia nigra, striatum, hippocampus, and internal capsule, with decreased cortical thickness in primary motor and somatosensory cortices. In the spinal cord, mSOD mice had significantly decreased volume of both the total grey and white matter; in the latter case, the volume change was confined to the dorsal white matter. Increased apoptosis, GFAP positive astrocytes, and/or activated microglia were observed in all those CNS regions that showed volume loss except for the hippocampus. The MRM findings in mSOD over expressing mice are similar to data previously obtained from a model of ALS parkinsonism dementia complex (ALS-PDC), in which neural damage occurred following a diet of washed cycad flour containing various neurotoxins. The primary difference between the two models involves a significantly greater decrease in spinal cord white matter volume in mSOD mice, perhaps reflecting variations in degeneration of the descending motor tracts. The extent to which several CNS structures are impacted in both murine models of ALS argues for a reevaluation of the nature of the pathogenesis of ALS since CNS structures involved in Parkinson's and Alzheimer's diseases appear to be affected as well. PMID- 17914181 TI - GnRH-mediated DAN production regulates the transcription of the GnRH receptor in gonadotrope cells. AB - The primary function of gonadotropin-releasing hormone (GnRH) is the regulation of pituitary gonadotropin hormone gene transcription, biosynthesis and release. These effects are mediated through intracellular mobilization of Ca2+ and activation of PKC isoforms and MAP kinases. We show here that DAN (differential screening-selected gene aberrative in neuroblastoma) which is a secreted bone morphogenic protein (BMP) antagonist belonging to the TGFbeta protein superfamily, is controlled by GnRH in murine gonadotrope cells. Acute GnRH stimulation induced a rapid, 27-fold, elevation of DAN mRNA, accompanied by an approximate 3-fold increase in the amount of mature DAN glycoprotein in the cell cytoplasm and in DAN secretion into the culture medium. Incubation of L beta T2 cells in DAN-containing medium altered the levels of a number of cellular proteins. Two of these were identified as the steroidogenic acute regulatory protein (StAR) and the actin-related protein 2/3 complex subunits 2 (p34-ARC) which are primarily involved in steroidogenesis and cytoskeleton remodelling, respectively. DAN caused an approximate 2-fold specific elevation in the cytoplasmic levels of both these proteins in L beta T2 cells. We further tested the effects of DAN on classical GnRH effects viz. gonadotropin and GnRH receptor gene expression. Co-transfection of L beta T2 cells with DAN and gonadotropin subunit promoter luciferase reporter genes had no effect on GnRH stimulation of alpha GSU and LH beta or on the additive GnRH and activin induction of FSH beta subunit transcription. However, co-transfection of DAN markedly inhibited the synergistic activation of GnRH and activin on GnRH receptor gene expression thus implicating DAN as a novel autocrine/paracrine factor that modulates GnRH function in pituitary gonadotropes. PMID- 17914182 TI - Effect of MTHFR polymorphisms on hyperhomocysteinemia in levodopa-treated Parkinsonian patients. AB - High plasma homocysteine levels have been observed in Parkinson's disease (PD) patients treated with levodopa. In this study, we investigated the effects of C677T and A1298C MTHFR polymorphisms, in association with L-DOPA daily dose and vitamin status, on hyperhomocysteinemia development in PD patients. Plasma homocysteine and folate/vitamin B12 levels were assayed in 49 L-DOPA-treated PD patients, and compared with those of 86 healthy subjects. Genotyping for MTHFR polymorphisms was carried out by DG-DGGE. Homocysteine levels were significantly higher in patients than in controls (16.3 +/- 5.7 vs. 11.7 +/- 2.7 micromol/l, P < 0.01). No significant differences were found between patients and controls with regard to folate/vitamin B12 levels, and MTHFR allele distribution. The TT+AA genotype was significantly more frequent in PD patients than in controls (32.5% vs. 17.4%, P < 0.05), but not associated with an increased risk for PD (OR = 2.3, CI = 1.0-5.2). Further, patients carrier of this genotype exhibited a mild hyperhomocysteinemia (22.1 +/- 4.9 micromol/l), while a protective effect was observed in patients having the CC+AA genotype (11.2 +/- 1.6 micromol/l; OR = 0.19, CI = 0.06-0.59). Interestingly, homocysteine levels were also moderately increased in patients with CT heterozygous genotype, in the context of either AA or AC (14.5 +/- 3.6 micromol/l), in comparison to subjects with the CC + AA genotype. Finally, we did not find any significant association of combined C677T and A1298C MTHFR polymorphisms with an increased risk for hyperhomocysteinemia in PD patients. A better understanding of the role of homocysteine and MTHFR genotypes in PD is needed to reveal novel approaches for disease management. PMID- 17914183 TI - Differential regulation of Smac/DIABLO and Hsp-70 during brain maturation. AB - The heat shock protein (Hsp) system is a cell defense mechanism constitutively expressed at the basal state and essential for cell survival in response to damaging stimuli. Apoptosis is a physiological cell death program that preserves tissue homeostasis. We investigated the intrinsic pathway of apoptosis at various stages of brain maturation in CD-1 mice, triggered by two mitochondrial proapoptotic proteins, cytochrome c and Smac/DIABLO, and the pathway's regulation by Hsp-70. Smac/DIABLO and Hsp-70 proteins were upregulated 2-fold and 1.5-3 fold, respectively, after birth. In contrast, in the presence of cytochrome c/2' deoxyadenosine 5'-triphosphate (dATP), caspase activity in mouse brain cell-free extracts increased 90-fold and 61-fold, at fetal and neonatal stages, whereas no activation was detected 15 days postnatally or at any subsequent times. These results indicate that the activation pattern of the intrinsic pathway of apoptosis undergoes a marked shift during postnatal maturation. PMID- 17914184 TI - Effects of dietary supplementation with N-acetyl cysteine, acetyl-L-carnitine and S-adenosyl methionine on cognitive performance and aggression in normal mice and mice expressing human ApoE4. AB - In addition to cognitive impairment, behavioral changes such as aggressive behavior, depression, and psychosis accompany Alzheimer's Disease. Such symptoms may arise due to imbalances in neurotransmitters rather than overt neurodegeneration. Herein, we demonstrate that combined administration of N acetyl cysteine (an antioxidant and glutathione precursor that protects against A beta neurotoxicity), acetyl-L-carnitine (which raises ATP levels, protects mitochondria, and buffers A beta neurotoxicity), and S-adenosylmethionine (which facilitates glutathione usage and maintains acetylcholine levels) enhanced or maintain cognitive function, and attenuated or prevented aggression, in mouse models of aging and neurodegeneration. Enhancement of cognitive function was rapidly reversed upon withdrawal of the formulation and restored following additional rounds supplementation. Behavioral abnormalities correlated with a decline in acetylcholine, which was also prevented by this nutriceutical combination, suggesting that neurotransmitter imbalance may contribute to their manifestation. Treatment with this nutriceutical combination was able to compensate for lack of dietary folate and vitamin E, coupled with administration of dietary iron as a pro-oxidant (which collectively increase homocysteine and oxidative damage to brain tissue), indicating that it provided antioxidant neuroprotection. Maintenance of neurotransmitter levels and prevention of oxidative damage underscore the efficacy of a therapeutic approach that utilizes a combination of neuroprotective agents. PMID- 17914185 TI - Pore-forming proteins share structural and functional homology with amyloid oligomers. AB - Degenerative diseases such as Alzheimer's, Parkinson's, and Huntington's diseases are believed to be causally related to the accumulation of amyloid oligomers that exhibit a common structure and may be toxic by a common mechanism involving permeabilization of membranes. We discovered that amyloid oligomers and the pore forming bacterial toxin, alpha-hemolysin (alpha HL), as well as human perforin from cytotoxic T lymphocytes, share a structural and functional homology at the level of their common reactivity with a conformation-dependent antibody that is specific for amyloid oligomers, A11. The alpha HL oligomeric pores and partially folded alpha HL protomer, but not the monomer alpha HL precursor reacts with A11 antibody. A11 antibody inhibits the hemolytic activity of alpha HL, indicating that the structural homology is functionally significant. Perforin oligomers were also recognized by A11. Amyloidogenic properties of alpha HL and perforin were confirmed spectroscopically and morphologically. These results indicate that pore forming proteins (PFP) and amyloid oligomers share structural homology and suggest that PFPs and amyloid oligomers share the same mechanism of membrane permeabilization. PMID- 17914186 TI - Reversible unilateral nigrostriatal pathway inhibition induced through expression of adenovirus-mediated clostridial light chain gene in the substantia nigra. AB - Clostridial light chain (LC) inhibits synaptic transmission by digesting a vesicle-docking protein, synaptobrevin, without killing neurons. We here report the feasibility of creating a rat hemiparkinsonism model through LC gene expression in the substantia nigra (SN), inhibiting nigrostriatal transmission. 40 adult Sprague Dawley rats were divided into four groups for SN injections of PBS, 6-hydroxydopamine (6-OHDA), or adenoviral vectors for the expression of LC (AdLC), or GFP (AdGFP). Amphetamine and apomorphine induced rotations were assessed before and after SN injection, revealing significant rotational alterations at 8 or 10 days after injection in both AdLC and 6-OHDA but not PBS and AdGFP groups. Induced rotation recovered by one month in AdLC rats but persisted in 6-OHDA rats. Histological analysis of the SN revealed LC and GFP expression with corresponding synaptobrevin depletion in the LC, but not the GFP groups. Tyrosine hydroxylase (TH) and dopamine transporter (DAT) immunohistochemistry (IHC) showed markedly decreased staining in ipsilateral SN and striatum in 6-OHDA but not AdLC or AdGFP rats. Similarly, compared with contralateral, ipsilateral striatal dopamine level only decreased in 6-OHDA but not AdLC, AdGFP, or PBS treated rats. Thus, LC expression induces nigral synaptobrevin depletion with resulting inhibition of nigrostriatal synaptic transmission. Unlike 6-OHDA, LC expression inhibits synaptic activity without killing neurons. This approach, therefore, represents a potentially reversible means of nigrostriatal pathway inhibition as a model for Parkinson's disease. Such a model might facilitate transient and controlled nigral inhibition for studying striatal recovery, dopaminergic re-innervation, and normalization of striatal receptors following the recovery of nigrostriatal transmission. PMID- 17914187 TI - The use of isobaric tag peptide labeling (iTRAQ) and mass spectrometry to examine rare, primitive hematopoietic cells from patients with chronic myeloid leukemia. AB - Chronic Myeloid Leukemia (CML) is a hematopoietic stem cell disease, associated with a t(9, 22) chromosomal translocation leading to formation of the BCR/ABL chimeric protein, which has an intrinsic tyrosine kinase activity. Recently, the BCR/ABL tyrosine kinase inhibitor imatinib mesylate (imatinib) has been successfully used clinically, although, disease relapse can still occur. The precise detail of the mechanism by which CML cells respond to imatinib is still unclear. We therefore systematically examined the effects of imatinib on the primitive CML cell proteome, having first established that the drug inhibits proliferation and induces increased apoptosis and differentiation. To define imatinib-induced effects on the CML proteome, we employed isobaric tag peptide labeling (iTRAQ) coupled to two-dimensional liquid chromatography/tandem mass spectrometry. Given the limited clinical material available, the isobaric tag approach identified a large population of proteins and provided relative quantification on four samples at once. Novel consequences of the action of imatinib were identified using this mass spectrometric approach. DEAD-box protein 3, heat shock protein 105 kDa, and peroxiredoxin-3 were identified as potential protein markers for response to imatinib. PMID- 17914188 TI - A system for the directed evolution of the insecticidal protein from Bacillus thuringiensis. AB - Theoretically, the activity of AB-type toxin molecules such as the insecticidal toxin (Cry toxin) from B. thuringiensis, which have one active site and two binding site, is improved in parallel with the binding affinity to its receptor. In this experiment, we tried to devise a method for the directed evolution of Cry toxins to increase the binding affinity to the insect receptor. Using a commercial T7 phage-display system, we expressed Cry1Aa toxin on the phage surface as fusions with the capsid protein 10B. These recombinant phages bound to a cadherin-like protein that is one of the Cry1Aa toxin receptors in the model target insect Bombyx mori. The apparent affinity of Cry1Aa-expressing phage for the receptor was higher than that of Cry1Ab-expressing phage. Phages expressing Cry1Aa were isolated from a mixed suspension of phages expressing Cry1Ab and concentrated by up to 130,000-fold. Finally, random mutations were made in amino acid residues 369-375 in domain 2 of Cry1Aa toxin, the mutant toxins were expressed on phages, and the resulting phage library was screened with cadherin like protein-coated beads. As a result, phages expressing abnormal or low affinity mutant toxins were excluded, and phages with high-affinity mutant toxins were selected. These results indicate that a method combining T7 phage display with selection using cadherin-like protein-coated magnetic beads can be used to increase the activity of easily obtained, low-activity Cry toxins from bacteria. PMID- 17914189 TI - Cloning, characterization, and transformation of the phosphoethanolamine N methyltransferase gene (ZmPEAMT1) in maize (Zea mays L.). AB - N-methylation of phosphoethanolamine, the committing step in choline (Cho) biosynthesis in plants, is catalyzed by S-adenosyl-L-methionine: phosphoethanolamine N-methyltransferase (PEAMT, EC 2.1.1.103). Herein we report the cloning and characterization of the novel maize phosphoethanolamine N methyltransferase gene (ZmPEAMT1) using a combination of bioinformatics and a PCR based allele mining strategy. The cDNA sequence of ZmPEAMT1 gene is 1,806 bp in length and translates a 495 amino acids peptide. The upstream promoter sequence of ZmPEAMT1 were obtained by TAIL-PCR, and contained four kinds of putative cis acting regulatory elements, including stress-responsive elements, phytohormone responsive elements, pollen developmental special activation elements, and light induced signal transduction elements, as well as several other structural features in common with the promoter of rice and Arabidopsis homologies. RT-PCR analysis showed that expression of ZmPEAMT1 was induced by salt stress and suppressed by high temperature. Over-expression of ZmPEAMT1 enhanced the salt tolerance, root length, and silique number in transgenic Arabidopsis. These data indicated that ZmPEAMT1 maybe involved in maize root development and stress resistance, and maybe having a potential application in maize genetic engineering. PMID- 17914190 TI - Naturally occurring catalytic antibodies: evidence for preferred development of the catalytic function in IgA class antibodies. AB - IgG class antibodies express catalytic activities rarely and at very low levels. Here, we studied polyclonal IgA and IgG preparations from healthy human sera and saliva for the ability to hydrolyze model peptidyl-aminomethylcoumarin (peptide AMC) substrates. These substrates permit objective evaluation of the catalytic potential of the antibody classes with minimal effects of noncovalent interactions occurring at sites remote from the reaction center. The IgA preparations hydrolyzed Glu-Ala-Arg-AMC at rates 3-orders of magnitude greater than IgG preparations from the same individuals. The cleavage occurred preferentially on the C terminal side of a basic residue. The activity was confirmed using monoclonal IgAs isolated from patients with multiple myeloma. Active site-directed inhibitors of serine proteases inhibited the catalytic activity and were bound irreversibly by the IgA, suggesting the involvement of a serine protease-like mechanism similar to that utilized by previously described IgM antibodies. These observations suggest that mechanisms underlying B cell clonal selection favor the retention and improvement of catalytic activity in the IgA, but not the IgG compartment of the immune response. PMID- 17914191 TI - Revealing differentially expressed proteins in two morphological forms of Spirulina platensis by proteomic analysis. AB - Spirulina is distinguished from other cyanobacteria by its spiral morphology; however, this cyanobacterium has frequently been observed with a linear morphology in laboratory and industrial conditions. In our laboratory conditions, the simultaneously presence of the linear and spiral forms has also been observed. In the present study, the two forms of S. platensis C1 were separated and grown as axenic cultures in order to study the proteins that were differentially expressed in the soluble and insoluble protein fractions of the spiral and the linear forms. Two dimensional-differential gel electrophoresis (2D DIGE) was performed to separate differentially expressed proteins that were subsequently identified by mass spectrometry. The differentially expressed proteins suggested two points. First, the morphological change is possibly induced by various environmental stresses such as oxygen level, carbon dioxide level, nutrient availability, and light. Second, the change of cell-shape might be a result of the change in a cell shape determination mechanism. Thus, this study is the first to show evidence at the protein level that may explain this morphological transformation in Spirulina. PMID- 17914193 TI - Construction of microarrays for genotyping of DQA using unmodified 45-mer oligonucleotide. AB - The human leukocyte antigen (HLA) class II system is strongly connected to immunological response and its compatibility between tissues is critical in transplantation. The simple robust typing analyses of HLA genes are extremely important. In this paper, we developed an approach based on microarray technology for genotyping of DQA gene. The microarrays were constructed with a total 31 unmodified 45-mer oligonucleotide. The second exon of DQA gene was amplified, and allowed to hybridize with the array. DQA genotypes were assigned by quantitative analysis of the hybridization results. The arrays were evaluated by DQA genotyping of nine reference samples and 120 clinical samples. The results demonstrate that the genotyping accuracy/concordance achieved 97.5% compared with the direct DNA sequencing. Although our methods did not perform high-resolution genotyping, it could be an alternative for serological typing in routine medical practice. PMID- 17914194 TI - On the stability of plasmid DNA vectors during cell culture and purification. AB - Gene therapy and DNA vaccination applications have increased the demand for highly purified plasmid DNA (pDNA) in the last years. One of the main problems related to the scale-up of pDNA purification is the degradation of the supercoiled (sc) isoforms during cell culture and multi-stage purification. In this work, a systematic study of the stability of two model plasmids (3,697 and 6,050 bp) during a mid-scale production process, which includes fermentation, alkaline lysis, isopropanol and ammonium sulphate precipitation and hydrophobic interaction chromatography, was performed. Results indicate that by extending cell culture (up to 26 h) and cell lysis (up to 2 h) it is possible to significantly reduce the amounts of RNA, without significantly compromising the yields of the sc pDNA isoform, a feature that could be conveniently exploited for downstream processing purposes. The stability of pDNA upon storage of E. coli pellets at different temperatures indicates that, differently from RNA, pDNA is remarkably stable when stored in cell pellets (>3 weeks at 4 degrees C, >12 weeks at -20 degrees C) prior to processing. With alkaline lysates, however, storage at -20 degrees C is mandatory to avoid sc pDNA degradation within the first 8 weeks. Furthermore, the subsequent purification steps could be carried out at room temperature without significant pDNA degradation. Since the unit operations and process conditions studied in this work are similar to those generally used for plasmid DNA production, the results presented here may contribute to improve the current knowledge on plasmid stability and process optimization. PMID- 17914195 TI - RNAi-mediated male sterility of tobacco by silencing TA29. AB - The superior performance of F1 hybrids has a significant impact on agricultural productivity. For commercial application, the availability of an efficient system for obtaining male-sterile lines of crops is an essential prerequisite. Here we have investigated the use of RNA interference (RNAi) technology to silence a male specific gene in the model host tobacco. TA29 is expressed exclusively in anthers at the time of microspore development. About 10 out of 13 tobacco lines transformed with a hairpin RNAi construct containing TA29 sequences were male sterile. Transgenic plants were phenotypically indistinguishable from non transgenic plants. At the anthesis stage, pollen grains from transgenic, male sterile plants were aborted and lysed in comparison to the round and fully developed pollen in non-transgenic plants. Microscopic analysis of anthers showed selective degradation of tapetum in transgenic plants with no microspore development. One week after self-pollination, the ovules of non-transgenic plants were double the size of those in transgenic plants, due to successful self fertilization. Male sterile transgenic plants set seed normally, when cross pollinated with pollen from non-transgenic plants, confirming no adverse effect on the female parts of the flower. These results show that silencing of male specific genes by RNAi is potentially a useful tool for generating male-sterile lines for producing hybrid seed. PMID- 17914192 TI - Exploiting dendritic cells for active immunotherapy of cancer and chronic infections. AB - Dendritic cells (DCs) are important antigen-presenting cells (APCs) that can prime naive T cells and control adaptive immune responses with respect to magnitude, memory and self-tolerance. Understanding the biology of these cells is central to the development of new generation immunotherapies for cancer and chronic infections. This review presents a brief overview of DC biology and of the preparation and use of DC-based vaccines. PMID- 17914197 TI - Each person has some capability or calling to improve upon what surrounds him. PMID- 17914196 TI - The targeted overexpression of a Claudin mutant in the epidermis of transgenic mice elicits striking epidermal and hair follicle abnormalities. AB - Skin is one of the largest organs of the body, and is formed during development through a highly orchestrated process involving mesenchymal-epithelial interactions, cell commitment, and terminal differentiation. It protects against microorganism invasion and UV irradiation, inhibits water loss, regulates body temperature, and is an important part of the immune system. Using transgenic mouse technology, we have demonstrated that Claudin (Cldn)-containing tight junctions (TJs) are intricately involved in cell signaling during epidermal differentiation and that an epidermal suprabasal overexpression of Cldn6 results in a perturbed epidermal terminal differentiation program with distinct phenotypic abnormalities. To delineate the role of the Cldn cytoplasmic tail domain in epidermal differentiation, we engineered transgenic mice targeting the overexpression of a Cldn6 cytoplasmic tail-truncation mutant in the epidermis. Transgenic mice were characterized by a lethal barrier dysfunction in addition to the existence of hyperproliferative squamous invaginations/cysts replacing hair follicles. Immunohistochemical analysis revealed an epidermal cytoplasmic accumulation of Cldn6, Cldn11, Cldn12, and Cldn18, downregulation of Cldn1 and aberrant expression of various classical markers of epidermal differentiation; namely the basal keratins as well as K1, involucrin, loricrin, and filaggrin. Collectively these studies suggest an important role for Cldns in epidermal/hair follicle differentiation programs likely involving cross talk to signaling pathways (e.g., Notch) directing cell fate selection and differentiation. PMID- 17914198 TI - Cataract surgery and spectacle independence. PMID- 17914199 TI - Corneal hysteresis following descemetorhexis with endokeratoplasty: early results. AB - The corneal biomechanical properties following descemetorhexis with endokeratoplasty (DXEK) and in normal subjects were studied in 100 eyes (12 DXEK and 88 age-matched normal subjects). Corneal hysteresis measurements were significantly lower in DXEK vs normal subjects. DXEK has a direct effect on corneal hysteresis and corneal biomechanical properties of the human cornea. PMID- 17914200 TI - Penetration of topically applied tacrolimus into the aqueous humor in Behcet's disease. AB - We investigated the concentration of tacrolimus in aqueous humor, and serum of patients with ocular Behcet's disease after topical application in 11 patients. All received 1 drop of isotonic tacrolimus solution 0.3% every 6 hours for 3 days. The mean tacrolimus concentration in the aqueous and serum, as measured by microparticle enzyme immunoassay, were 12.49 and 0.76 ng/mL, respectively. Tacrolimus may be a promising treatment modality in anterior uveitis in Behcet's disease by topical application. PMID- 17914201 TI - Effect of topical cyclosporine on tear functions in tear-deficient dry eyes. AB - We evaluated the efficacy of topical 2% cyclosporine drops in the treatment of tear-deficient dry eye because of acquired primary lachrymal disease (APLD; 15 patients) and Sjogren syndrome (SS; 15 patients). Symptoms of dryness tended to improve in patients with SS. Schirmer score improved in patients with APLD. Topical CsA 2% drops appears to be safe and effective in the treatment of dry eye patients because in patients with APLD and SS, there is trend toward improvement. PMID- 17914202 TI - Visual outcome after cataract surgery in extremely high axial myopia. AB - We retrospectively investigated visual outcome after cataract surgery with implantation of zero or negative-power intraocular lenses (IOL) in 110 eyes with extremely high axial myopia. Extremely high axial myopes can benefit from cataract surgery and implantation of zero or negative-power IOLs with few complications. Factors associated with visual outcome included preoperative best corrected visual acuity, gender, and age. Method of surgery did not influence the visual outcome. PMID- 17914203 TI - The preventive effect of capsular tension ring in phacoemulsification of senile cataracts with pseudoexfoliation. AB - We evaluated the preventive role of the capsular tension ring (CTR) in zonular and capsular complications in the phacoemulsification surgery of cataracts with pseudoexfoliation (PEX) in two groups of eyes with senile cataract associated with PEX, with (Group A) or without (Group B) CTR implantation. CTR implantation did not prove as effective as expected in preventing the zonulo-capsular complications. Contradictory to the significant increase of intracapsular intraocular lens (IOL) implantation rate, post-operative visual acuity did not improve in the same value. PMID- 17914204 TI - Comparing dynamic contour tonometry to Goldmann and hand-held tonometry in normal, ocular hypertension, and glaucoma populations. AB - We prospectively compared dynamic contour tonometry (DCT) to Goldmann (GAT) and hand-held tonometry (HHT) in normal, ocular hypertension, and glaucoma populations. Both measurements were made on each patient within a 5-minute period during routine office exams over 4 months. While DCT is in good overall agreement with GAT and HTT, there is some systematic deviation at different pressure ranges in normal, ocular hypertension, and glaucoma populations. PMID- 17914205 TI - Large subretinal hemorrhage after intravitreal bevacizumab (Avastin) for age related macular degeneration. AB - We report the development of a large subretinal hemorrhage 4 weeks after intravitreal bevacizumab in an 84-year-old male with 20/70 visual acuity caused by occult choroidal neovascularization from neovascular age-related macular degeneration in one eye that was treated with 1.25 mg of intravitreal bevacizumab. PMID- 17914206 TI - Wolfram syndrome: case report and review of the literature. AB - Wolfram syndrome (WFS) is a rare diffuse neurodegenerative disorder characterized by diabetes insipidus, diabetes mellitus, optic atrophy, deafness, and a wide variety of central nervous system abnormalities. Insulin-dependent diabetes mellitus with optic nerve atrophy is sufficient criteria for the diagnosis. WFS is a devastating disease for the patients and their families. This study emphasizes the need for careful evaluation of cases having insulin-dependent diabetes mellitus and optic atrophy. PMID- 17914207 TI - Acute retinal necrosis complicating chicken pox in a healthy adult: a case report and review of literature. AB - We report a case of unilateral acute retinal necrosis (ARN) with marked vitritis and retinal necrosis leading to retinal breaks following chicken pox successfully treated with intravenous acyclovir followed by oral acyclovir, orbital floor triamcinolone injections to contain the inflammation, and barrier laser therapy to secure the retinal breaks with good visual outcome. This case is unusual in its severity and the novel use orbital floor triamcinolone therapy to contain ARN inflammation. PMID- 17914208 TI - Involution of endophytic optic disc hemangioma with a single session of photodynamic treatment. AB - We report a 58-year-old otherwise healthy woman with unilateral endophytic optic disc hemangioma and associated serous retinal detachment and exudation. A standard verteporfin dose was given and a photodynamic treatment (PDT) protocol was followed. A moderate vitreous hemorrhage occurred four days after PDT. The hemangioma regressed markedly 6 weeks after the procedure and serous retinal detachment showed marked resolution. PMID- 17914209 TI - Primary conjunctival herpetic geographic ulcer in an immunocompetent patient. AB - Herpes simplex virus ocular infections are a major cause of corneal blindness in the developed world. Signs of a primary infection are limited to the lids, conjunctiva and cornea. The presence of a conjunctival dendrite without corneal involvement has been reported. Although conjunctival ulceration has been associated with recurrent attacks, it has not been documented as a manifestation of primary herpetic infection. We report a case of primary herpetic infection with geographic conjunctival ulcer with multiple corneal dendrites. PMID- 17914210 TI - Amniotic membrane transplantation in ocular rosacea. AB - A 40-year-old patient with the diagnosis of acne rosacea presented with perforated peripheral corneal ulcer. Three applications of cyanoacrylate glue along with bandage contact lens failed to seal the perforation. Two months after the presentation, corneal perforation was successfully managed with amniotic membrane transplantation. The final best corrected visual acuity of 6/6 was achieved. PMID- 17914211 TI - Congenital isolate and total optic disc coloboma: case report and review of the literature. AB - Isolated and total optic disc coloboma is a markedly rare congenital anomaly of the eye. Congenital eye colobomas frequently present with eye anomalies and systemic signal findings. We present a case of isolated total optic disc coloboma without systemic involvement, which has been rarely reported in the literature. PMID- 17914213 TI - Zinc supplementation or regulation of its homeostasis: advantages and threats. AB - To accomplish its multifunctional biological roles, zinc requires precise homeostatic mechanisms. There are efficient mechanisms that regulate zinc absorption from the alimentary tract and its excretion by the kidney depending on the organism demands. The regulatory mechanisms of cellular zinc inflow, distribution, and zinc outflow are so efficient that symptoms of zinc deficiency are rare, and symptoms connected with its massive accumulation are even more rare. The efficiency of homeostatic mechanisms that prevent zinc deficiency or excessive zinc accumulation in the organism is genetically conditioned. It seems that an essential element of zinc homeostasis is the efficiency of zinc transmembrane exchange mechanisms. Intracellular free zinc concentration is higher than in extracellular space. Physiologically, the active outflow of zinc ions from the cell depends on the increase of its concentration in extracellular space. The ion pumps activity depends on the efficiency by which the cell manages energy. Considering the fact that zinc deficiency accelerates apoptosis and that excessive zinc accumulation inside cells results in a toxic effect that forces its death brings about several questions: Is intensification and acceleration of changes in zinc metabolism with age meaningful? Is there a real zinc deficiency occurring with age or in connection with the aforementioned pathological processes, or is it just a case of tissue and cell redistribution? When discussing factors that influence zinc homeostasis, can we consider zinc supplementation or regulation of zinc balance in the area of its redistribution? To clarify these aspects, an essential element will also be the clear understanding of the nomenclature used to describe changes in zinc balance. Zinc homeostasis can be different in different age groups and depends on sex, thus zinc dyshomeostasis refers to changes in its metabolism that deviate from the normal rates for a particular age group and sex. This concept is very ample and implies that zinc deficiency may result from a low-zinc diet, poor absorption, excessive loss of zinc, zinc redistribution in intra- and extracellular compartments, or a combination of these factors that is inadequate for the given age and sex group. Such factor or factors need to be considered for preventing particular homeostasis disorders (or dyshomeostasis). Regulation of zinc metabolism by influencing reversal of redistribution processes ought to be the main point of pharmacologic and nonpharmacologic actions to reestablish zinc homeostasis. Supplementation and chelation are of marginal importance and can be used to correct long-term dietary zinc deficiency or zinc poisoning or in some cases in therapeutic interventions. In view of its biological importance, the problem posed by the influence of zinc metabolism requires further investigation. To date, one cannot consider, for example, routine zinc supplementation in old age, because changes of metabolism with age are not necessarily a cause of zinc deficiency. Supplementation is warranted only in cases in which deficiency has been established unambiguously. An essential element is to prevent sudden changes in zinc metabolism, which lead to dyshomeostasis in the terms defined here. The primary prophylaxes, regular physical activity, efficient treatment of chronic diseases, are all elements of such prevention. PMID- 17914214 TI - Cadmium, lead, and selenium in cord blood and thyroid hormone status of newborns. AB - Cadmium (Cd), lead (Pb), and selenium (Se) concentrations in cord whole blood, sampled from 24 women at the time of delivery in a hospital in Tokyo in 2005, were determined by inductively coupled plasma mass spectrometry with a reaction cell. Signal enhancement caused by nonspectroscopic interference for Se was evident and the standard addition technique was essential for correcting the interference. Median concentration in cord bloods was 0.20 ng/g, 6.7 ng/g (0.67 microg/dL), and 191 ng/g for Cd, Pb and Se, respectively. Lead concentration was lower, whereas Se concentration was higher, than those reported in other countries. The trace element concentration was related to the levels of thyroid stimulating hormone (TSH) and free thyroxin (fT4) in the neonatal blood sampled at 4-6 days postpartum. A significantly negative correlation was observed between Cd concentrations in cord blood and TSH concentration in neonatal blood. The result indicated the possible effect of in utero Cd exposure on thyroid hormone status of newborns and that Cd exposure level should be assessed as a covariate in the survey on the relationship between in utero chemicals (e.g., PCBs) exposure and thyroid hormone status. PMID- 17914216 TI - Urinary fluoride reference values determined by a fluoride ion selective electrode. AB - As fluoride has a very short half-life in the body and the major route for fluoride excretion is via the kidney, human exposure is best measured in urine, where the concentration is expected to be highest. The urinary fluoride concentrations of 167 healthy Japanese adults were determined by means of a fluoride ion selective electrode. When the results were corrected for a specific gravity rho = 1.024 g cm-3, the histogram of urinary fluoride concentrations highly skewed toward low values with sharp peakedness (skewness = 1.56, kurtosis = 3.08). The normality of the log-transformed histogram (skewness = 0.12, kurtosis = 0.07) and the straight line on log-probability paper clearly showed a key feature of lognormal distribution of urinary fluoride. A geometric mean (GM) of 613.8 microg/l and 95% confidential interval (CI) of 241.0-1633.1 microg/l were established as reference values for urinary fluoride. The results presented in this study will be useful as guidelines for the biological monitoring of fluoride in normal subjects and individuals at risk of occupational or environmental fluoride exposure. PMID- 17914215 TI - Serum zinc concentrations in cystic fibrosis patients aged above 4 years: a cross sectional evaluation. AB - AIM: Assess the risk of zinc (Zn) deficiency in the older cystic fibrosis (CF) population. METHOD: Cross-sectional investigation of all CF patients above the age of 4 followed at the Ghent University center between 2002 and 2003. Data on age, weight, height z-score, pancreatic and pulmonary functions, chronic Pseudomonas infection, and CF transmembrane conductance regulator (CFTR) mutations were collected. Serum Zn, vitamins (vit) A and E, retinol-binding protein (RBP), albumin, sedimentation rate, total IgG, and cholesterol were determined. Serum Zn was compared with a local healthy control group (Van Biervliet et al., Biol Trace Elem Res 94:33-40, 2003) and with literature data (Hotz C, et al. Am J Clin Nutr 78:756-764, 2003). RESULTS: 101 patients (median age 16 years) were included. There was no difference in serum Zn concentration between CF patients and controls. In CF patients no difference in serum Zn concentration between pancreatic-sufficient or pancreatic-insufficient patients was seen. Serum Zn was not associated to nutritional status or height z-score. A significant association serum Zn to serum albumin (p < 0.0005) and to vit A (p < 0.01) was seen. No associations of serum Zn to serum vit E, RBP, cholesterol, or CFTR were present, but there is a significant association serum Zn to forced vital capacity (p < 0.01). Serum Zn was not associated to inflammatory parameters or chronic Pseudomonas infection. CONCLUSION: Comparison of CF patients with local controls revealed no significant differences. However, because persisting steatorrhea increases Zn loss (Easley et al., J Pediatr Gastroenterol Nutr 26:136 139, 1998) and 12.6% of our population has a serum Zn below the p value of 2.5 of the NHANES II study (Hotz C, et al. Am J Clin Nutr 78:756-764, 2003), there could remain an increased risk of Zn deficiency in some CF patients. Furthermore, the association with pulmonary function needs more investigation. PMID- 17914217 TI - Defensive role of quercetin against imbalances of calcium, sodium, and potassium in galactosemic cataract. AB - Galactosemic cataracts are characterized by electrolyte disturbances resulting in osmotic imbalance and loss of transparency. We have studied the defensive role of quercetin, a bioflavonoid, against the alterations of calcium (Ca2+), sodium (Na+), and potassium (K+) concentrations in galactose-induced cataract in a rodent model. The experimental study was conducted on weanling male Wistar rats with an average body weight of 34 +/- 0.9 g. Different groups received normal stock AIN 93 diet (group A, n = 8), AIN 93 diet with 30% galactose (group B, n = 8), and AIN 93 diet with 30% galactose + quercetin at 400 mg/100 g diet (group C, n = 8). Aldose reductase activity and protein content and concentrations of Ca2+, Na+, and K+ were determined in normal and cataractous lenses. Treatment with quercetin resulted in a significant decrease in Na+ and Ca2+ and aldose reductase levels and an increase in K+ and protein levels in galactosemic cataractous lenses. These results imply that inclusion of quercetin contributes to lens transparency through the maintenance of characteristic osmotic ion equilibrium and protein levels of the lens. PMID- 17914218 TI - Efficacy of chromium(III) supplementation on growth, body composition, serum parameters, and tissue chromium in rats. AB - Chromium(III) is often claimed to have a positive effect on body composition, while the responses in researches with supplementation of different chemical form of chromium are various and inconsistent. We have studied the effects of 6 weeks of treatment with three different forms of chromium (300 mug/kg) as chromium chloride, chromium tripicolinate, and chromium nanocomposite (CrNano) on growth, body composition, serum parameters, and tissue chromium in rats. The supplementation of CrNano significantly increased average daily gain, food efficiency, and lean body mass and decreased fat mass and body fat proportion and serum levels of glucose, urea nitrogen, triglyceride, and insulin. Chromium contents in liver, kidney, and hind leg muscle were increased significantly with the addition of CrNano in diet. The results indicate that chromium nanocomposite has higher efficacy on growth and body composition compared to the traditional chromium agents. PMID- 17914219 TI - Effects of artichoke extract supplementation on gonads of cadmium-treated rats. AB - The present study was designed to determine whether artichoke (Cynara scolymus) exerts a protective effect on gonads of cadmium-treated rats and if there is a relationship between artichoke supplementation and nitric oxide (NO) formation in cells. Forty Wistar albino male rats, weighing an average of 90 g each, were equally divided into four groups receiving 1 mg/100 g cadmium chloride by injection (group 1), the same dose CdCl2 plus 3 mg/100 g artichoke extract (group 2), the same dose of artichoke extract (group 3), and male controls (group 4). Four additional groups, labeled 5-8, consisted of identically treated and control female rats. After 4 weeks of treatment, the animals were killed and their gonads were removed for histological examination. As expected, the seminiferous tubules and Leydig cells were damaged by cadmium. Ovarian tissue was not damaged to the same extent as testicular cells. Artichoke extract exerted a clear protective effect against Cd-induced testicular damage and lowered NO production to the same level of that in the control groups. PMID- 17914220 TI - Bioenergetic investigation of action of lithium to Tetrahymena thermophila bF5 by microcalorimetry. AB - Microcalorimetry was employed to investigate the action of Li(I) to aquatic ecosystem from the point view of bioenergetics. Tetrahymena thermophila BF5 was chosen as the model organism. The power-time curves of T. thermophila BF5 growth metabolism in the absence and presence of Li(I) were obtained. The corresponding thermokinetic parameters were derived. The generation time was calculated as 592.3 min, which was consistent with the biomass values. Low concentration of Li(I) (1-20 mmol l-1) stimulated the growth of T. thermophila BF5, whereas the inhibition effect was observed in high concentration (30-100 mmol l-1). The value of IC50 was 52.8 mmol l-1. In the concentration range of 30-100 mmol l-1, the growth rate constants (k) and the maximum heat out power (P max) decrease with the concentration of Li(I), whereas the heat output (Q) increases slightly compared to the control. Other than the classic mechanism of inositol-phosphate cycle, the involvement of mitochondria mechanism was discussed and suggested. PMID- 17914221 TI - Effects of Nanoanatase TiO2 on photosynthesis of spinach chloroplasts under different light illumination. AB - With a photocatalyzed characteristic, nanoanatase TiO2 under light could cause an oxidation-reduction reaction. Our studies had proved that nano-TiO2 could promote photosynthesis and greatly improve spinach growth. However, the mechanism of nano TiO2 on promoting conversion from light energy to electron energy and from electron energy to active chemistry energy remains largely unclear. In this study, we report that the electron transfer, oxygen evolution, and photophosphorylation of chloroplast (Chl) from nanoanatase-TiO2-treated spinach were greatly increased under visible light and ultraviolet light illumination. It was demonstrated that nanoanatase TiO2 could greatly improve whole chain electron transport, photoreduction activity of photosystem II, O2-evolving and photophosphorylation activity of spinach Chl not only under visible light, but also energy-enriched electron from nanoanatase TiO2, which entered Chl under ultraviolet light and was transferred in photosynthetic electron transport chain and made NADP+ be reduced into NADPH, and coupled to photophosphorylation and made electron energy be transformed to ATP. Moreover, nanoanatase h+, which photogenerated electron holes, captured an electron from water, which accelerated water photolysis and O2 evolution. PMID- 17914222 TI - The improvement of spinach growth by nano-anatase TiO2 treatment is related to nitrogen photoreduction. AB - The improvement of spinach growth is proved to relate to N2 fixation by nano anatase TiO2 in this study. The results show that all spinach leaves kept green by nano-anatase TiO2 treatment and all old leaves of control turned yellow white under culture with N-deficient solution. And the fresh weight, dry weight, and contents of total nitrogen, NH4(+), chlorophyll, and protein of spinach by nano anatase TiO2 treatment presented obvious enhancement compared with control. Whereas the improvements of yield of spinach were not as good as nano-anatase TiO2 treatment under N-deficient condition, confirming that nano-anatase TiO2 on exposure to sunlight could chemisorb N2 directly or reduce N2 to NH3 in the spinach leaves, transforming into organic nitrogen and improving the growth of spinach. Bulk TiO2 effect, however, was not as significant as nano-anatase TiO2. A possible metabolism of the function of nano-anatase TiO2 reducing N2 to NH3 was discussed. PMID- 17914223 TI - The level of selenium and some other trace elements in different Libyan arable soils using instrumental neutron activation analysis. AB - Elemental analysis of soils from two different arable regions in Libya was carried out to measure the level of many trace elements. Instrumental neutron activation analysis was used for the determination of 10 elements, viz., (Ba, Ce, Co, Cr, Cs, Fe, Sc, Se, Th, and Zn), using their long-lived radionuclides. The accuracy of the measurements has been evaluated by analyzing two IAEA soil reference materials: IAEA Soil-7 and IAEA leak sediment SL-1; precision has been estimated by triplicate analysis of the sample and that of the reference material. Irradiations were carried out at the Tajura Research Center reactor, at 5-MW power level. It is clear that in the Libyan soil selenium concentration is somewhat lower than in other countries. The results show that trace metal concentrations in Libyan clay surface soil are higher than the sandy soil. PMID- 17914225 TI - Mechanism of DNA-binding loss upon single-point mutation in p53. AB - Over 50% of all human cancers involve p53 mutations,which occur mostly in the sequence-specific DNA-binding central domain (p53c), yielding little/non detectable af?nity to the DNA consensus site. Despite our current understanding of protein-DNA recognition,the mechanism(s) underlying the loss in protein-DNA binding afnity/ specificity upon single-point mutation are not well understood. Our goal is to identify the common factors governing the DNA-binding loss of p53c upon substitution of Arg 273 to His or Cys,which are abundant in human tumours. By computing the free energies of wild-type and mutant p53c binding to DNA and decomposing them into contributions from individual residues, the DNA-binding loss upon charge/noncharge -conserving mutation of Arg 273 was attributed not only to the loss of DNA phosphate contacts, but also to longer-range structural changes caused by the loss of the Asp 281 salt-bridge. The results herein and in previous works suggest that Asp 281 plays a critical role in the sequence specific DNA-binding function of p53c by (i)orienting Arg 273 and Arg 280 in an optimal position to interact with the phosphate and base groups of the consensus DNA, respectively, and (ii) helping to maintain the proper DNA-binding protein conformation. PMID- 17914224 TI - Theoretical analysis of noncanonical base pairing interactions in RNA molecules. AB - Noncanonical base pairs in RNA have strong structural and functional implications but are currently not considered for secondary structure predictions. We present results of comparative ab initio studies of stabilities and interaction energies for the three standard and 24 selected unusual RNA base pairs reported in the literature. Hydrogen added models of isolated base pairs, with heavy atoms frozen in their 'away from equilibrium' geometries, built from coordinates extracted from NDB, were geometry optimized using HF/6-31G** basis set, both before and after unfreezing the heavy atoms. Interaction energies, including BSSE and deformation energy corrections, were calculated, compared with respective single point MP2 energies, and correlated with occurrence frequencies and with types and geometries of hydrogen bonding interactions. Systems having two or more N-H...O/N hydrogen bonds had reasonable interaction energies which correlated well with respective occurrence frequencies and highlighted the possibility of some of them playing important roles in improved secondary structure prediction methods. Several of the remaining base pairs with one N-H...O/N and/or one C-H...O/N interactions respectively, had poor interaction energies and negligible occurrences. High geometry variations on optimization of some of these were suggestive of their conformational switch like characteristics. PMID- 17914226 TI - Incorporating evolution of transcription factor binding sites into annotated alignments. AB - Identifying transcription factor binding sites (TFBSs) is essential to elucidate putative regulatory mechanisms. A common strategy is to combine cross-species conservation with single sequence TFBS annotation to yield "conserved TFBSs". Most current methods in this field adopt a multi-step approach that segregates the two aspects. Again, it is widely accepted that the evolutionary dynamics of binding sites differ from those of the surrounding sequence. Hence, it is desirable to have an approach that explicitly takes this factor into account. Although a plethora of approaches have been proposed for the prediction of conserved TFBSs, very few explicitly model TFBS evolutionary properties, while additionally being multi-step. Recently, we introduced a novel approach to simultaneously align and annotate conserved TFBSs in a pair of sequences. Building upon the standard Smith-Waterman algorithm for local alignments, SimAnn introduces additional states for profiles to output extended alignments or annotated alignments. That is, alignments with parts annotated as gaplessly aligned TFBSs (pair-profile hits)are generated. Moreover,the pair- profile related parameters are derived in a sound statistical framework. In this article, we extend this approach to explicitly incorporate evolution of binding sites in the SimAnn framework. We demonstrate the extension in the theoretical derivations through two position-specific evolutionary models, previously used for modelling TFBS evolution. In a simulated setting, we provide a proof of concept that the approach works given the underlying assumptions,as compared to the original work. Finally, using a real dataset of experimentally verified binding sites in human mouse sequence pairs,we compare the new approach (eSimAnn) to an existing multi step tool that also considers TFBS evolution. Although it is widely accepted that binding sites evolve differently from the surrounding sequences, most comparative TFBS identification methods do not explicitly consider this.Additionally, prediction of conserved binding sites is carried out in a multi-step approach that segregates alignment from TFBS annotation. In this paper, we demonstrate how the simultaneous alignment and annotation approach of SimAnn can be further extended to incorporate TFBS evolutionary relationships. We study how alignments and binding site predictions interplay at varying evolutionary distances and for various profile qualities. PMID- 17914227 TI - Identification and annotation of promoter regions in microbial genome sequences on the basis of DNA stability. AB - Analysis of various predicted structural properties of promoter regions in prokaryotic as well as eukaryotic genomes had earlier indicated that they have several common features,such as lower stability, higher curvature and less bendability, when compared with their neighboring regions. Based on the difference in stability between neighboring upstream and downstream regions in the vicinity of experimentally determined transcription start sites, a promoter prediction algorithm has been developed to identify prokaryotic promoter sequences in whole genomes. The average free energy (E) over known promoter sequences and the difference (D) between E and the average free energy over the entire genome (G)are used to search for promoters in the genomic sequences. Using these cutoff values to predict promoter regions across entire Escherichia coli genome,we achieved a reliability of 70% when the predicted promoters were cross verified against the 960 transcription start sites (TSSs) listed in the Ecocyc database. Annotation of the whole E.coli genome for promoter region could be carried out with 49% accuracy. The method is quite general and it can be used to annotate the promoter regions of other prokaryotic genomes. PMID- 17914228 TI - Parsing regulatory DNA: general tasks, techniques, and the PhyloGibbs approach. AB - In this review, we discuss the general problem of understanding transcriptional regulation from DNA sequence and prior information. The main tasks we discuss are predicting local regions of DNA, cis-regulatory modules (CRMs) that contain binding sites for transcription factors (TFs), and predicting individual binding sites. We review various existing methods, and then describe the approach taken by PhyloGibbs, a recent motif-finding algorithm that we developed to predict TF binding sites, and PhyloGibbs-MP, an extension to PhyloGibbs that tackles other tasks in regulatory genomics, particularly prediction of CRMs. PMID- 17914229 TI - Evolutionary insights from suffix array-based genome sequence analysis. AB - Gene and protein sequence analyses, central components of studies in modern biology are easily amenable to string matching and pattern recognition algorithms. The growing need of analysing whole genome sequences more efficiently and thoroughly, has led to the emergence of new computational methods. Suffix trees and suffix arrays are data structures, well known in many other areas and are highly suited for sequence analysis too. Here we report an improvement to the design of construction of suffix arrays. Enhancement in versatility and scalability, enabled by this approach, is demonstrated through the use of real life examples. The scalability of the algorithm to whole genomes renders it suitable to address many biologically interesting problems. One example is the evolutionary insight gained by analysing unigrams, bi-grams and higher n-grams, indicating that the genetic code has a direct influence on the overall composition of the genome. Further, different proteomes have been analysed for the coverage of the possible peptide space, which indicate that as much as a quarter of the total space at the tetra-peptide level is left un-sampled in prokaryotic organisms, although almost all tri-peptides can be seen in one protein or another in a proteome. Besides, distinct patterns begin to emerge for the counts of particular tetra and higher peptides, indicative of a 'meaning' for tetra and higher n-grams. The toolkit has also been used to demonstrate the usefulness of identifying repeats in whole proteomes efficiently. As an example, 16 members of one COG,coded by the genome of Mycobacterium tuberculosis H37Rv have been found to contain a repeating sequence of 300 amino acids. PMID- 17914231 TI - Protein mechanics: a route from structure to function. AB - In order to better understand the mechanical properties of proteins, we have developed simulation tools which enable these properties to be analysed on a residue-by-residue basis. Although these calculations are relatively expensive with all-atom protein models, good results can be obtained much faster using coarse-grained approaches. The results show that proteins are surprisingly heterogeneous from a mechanical point of view and that functionally important residues often exhibit unusual mechanical behaviour. This finding offers a novel means for detecting functional sites and also potentially provides a route for understanding the links between structure and function in more general terms. PMID- 17914230 TI - A method for computing the inter-residue interaction potentials for reduced amino acid alphabet. AB - Inter-residue potentials are extensively used in the design and evaluation of protein structures. However,dealing with all (20 x 20) interactions becomes computationally difficult in extensive investigations. Hence, it is desirable to reduce the alphabet of 20 amino acids to a smaller number. Currently, several methods of reducing the residue types exist; however a critical assessment of these methods is not available. Towards this goal,here we review and evaluate different methods by comparing with the complete (20 x 20) matrix of Miyazawa Jernigan potential, including a method of grouping adopted by us, based on multi dimensional scaling (MDS). The second goal of this paper is the computation of inter-residue interaction energies for the reduced amino acid alphabet, which has not been explicitly addressed in the literature until now. By using a least squares technique, we present a systematic method of obtaining the interaction energy values for any type of grouping scheme that reduces the amino acid alphabet. This can be valuable in designing the protein structures. PMID- 17914232 TI - Protein local conformations arise from a mixture of Gaussian distributions. AB - The classical approaches for protein structure prediction rely either on homology of the protein sequence with a template structure or on ab initio calculations for energy minimization. These methods suffer from disadvantages such as the lack of availability of homologous template structures or intractably large conformational search space, respectively. The recently proposed fragment library based approaches first predict the local structures,which can be used in conjunction with the classical approaches of protein structure prediction. The accuracy of the predictions is dependent on the quality of the fragment library. In this work, we have constructed a library of local conformation classes purely based on geometric similarity. The local conformations are represented using Geometric Invariants, properties that remain unchanged under transformations such as translation and rotation, followed by dimension reduction via principal component analysis. The local conformations are then modeled as a mixture of Gaussian probability distribution functions (PDF). Each one of the Gaussian PDF's corresponds to a conformational class with the centroid representing the average structure of that class. We find 46 classes when we use an octapeptide as a unit of local conformation. The protein 3-D structure can now be described as a sequence of local conformational classes. Further, it was of interest to see whether the local conformations can be predicted from the amino acid sequences. To that end,we have analyzed the correlation between sequence features and the conformational classes. PMID- 17914233 TI - Exploring conformational space using a mean field technique with MOLS sampling. AB - The computational identification of all the low energy structures of a peptide given only its sequence is not an easy task even for small peptides,due to the multiple-minima problem and combinatorial explosion. We have developed an algorithm, called the MOLS technique,that addresses this problem, and have applied it to a number of different aspects of the study of peptide and protein structure. Conformational studies of oligopeptides, including loop sequences in proteins have been carried out using this technique. In general the calculations identified all the folds determined by previous studies,and in addition picked up other energetically favorable structures. The method was also used to map the energy surface of the peptides. In another application, we have combined the MOLS technique, using it to generate a library of low energy structures of an oligopeptide, with a genetic algorithm to predict protein structures. The method has also been applied to explore the conformational space of loops in protein structures.Further, it has been applied to the problem of docking a ligand in its receptor site, with encouraging results. PMID- 17914235 TI - Use of secondary structural information and C alpha-C alpha distance restraints to model protein structures with MODELLER. AB - Protein secondary structure predictions and amino acid long range contact map predictions from primary sequence of proteins have been explored to aid in modelling protein tertiary structures. In order to evaluate the usefulness of secondary structure and 3D-residue contact prediction methods to model protein structures we have used the known Q3 (alpha-helix,beta-strands and irregular turns/loops) secondary structure information, along with residue-residue contact information as restraints for MODELLER. We present here results of our modelling studies on 30 best resolved single domain protein structures of varied lengths. The results shows that it is very difficult to obtain useful models even with 100% accurate secondary structure predictions and accurate residue contact predictions for up to 30% of residues in a sequence. The best models that we obtained for proteins of lengths 37, 70, 118, 136 and 193 amino acid residues are of RMSDs 4.17, 5.27, 9.12, 7.89 and 9.69,respectively. The results show that one can obtain better models for the proteins which have high percent of alpha-helix content. This analysis further shows that MODELLER restrain optimization program can be useful only if we have truly homologous structure(s) as a template where it derives numerous restraints, almost identical to the templates used. This analysis also clearly indicates that even if we satisfy several true residue residue contact distances, up to 30%of their sequence length with fully known secondary structural information, we end up predicting model structures much distant from their corresponding native structures. PMID- 17914234 TI - Analysis on sliding helices and strands in protein structural comparisons: a case study with protein kinases. AB - Protein structural alignments are generally considered as 'golden standard' for the alignment at the level of amino acid residues. In this study we have compared the quality of pairwise and multiple structural alignments of about 5900 homologous proteins from 718 families of known 3-D structures. We observe shifts in the alignment of regular secondary structural elements (helices and strands) between pairwise and multiple structural alignments. The differences between pairwise and multiple structural alignments within helical and beta-strand regions often correspond to 4 and 2 residue positions respectively. Such shifts correspond approximately to "one turn" of these regular secondary structures. We have performed manual analysis explicitly on the family of protein kinases. We note shifts of one or two turns in helix-helix alignments obtained using pairwise and multiple structural alignments. Investigations on the quality of the equivalent helix-helix, strand-strand pairs in terms of their residue side-chain accessibilities have been made. Our results indicate that the quality of the pairwise alignments is comparable to that of the multiple structural alignments and, in fact, is often better. We propose that pairwise alignment of protein structures should also be used in formulation of methods for structure prediction and evolutionary analysis. PMID- 17914236 TI - ARC: automated resource classifier for agglomerative functional classification of prokaryotic proteins using annotation texts. AB - Functional classification of proteins is central to comparative genomics. The need for algorithms tuned to enable integrative interpretation of analytical data is felt globally. The availability of a general,automated software with built-in flexibility will significantly aid this activity. We have prepared ARC (Automated Resource Classifier), which is an open source software meeting the user requirements of flexibility. The default classification scheme based on keyword match is agglomerative and directs entries into any of the 7 basic non overlapping functional classes: Cell wall, Cell membrane and Transporters (C), Cell division (D), Information (I), Translocation (L), Metabolism (M), Stress(R), Signal and communication (S) and 2 ancillary classes: Others (O) and Hypothetical (H). The keyword library of ARC was built serially by first drawing keywords from Bacillus subtilis and Escherichia coli K12. In subsequent steps,this library was further enriched by collecting terms from archaeal representative Archaeoglobus fulgidus, Gene Ontology, and Gene Symbols. ARC is 94.04% successful on 6,75,663 annotated proteins from 348 prokaryotes. Three examples are provided to illuminate the current perspectives on mycobacterial physiology and costs of proteins in 333 prokaryotes. ARC is available at http://arc.igib.res.in. PMID- 17914237 TI - Synonymous codon usage in different protein secondary structural classes of human genes: implication for increased non-randomness of GC3 rich genes towards protein stability. AB - The relationship between the synonymous codon usage and different protein secondary structural classes were investigated using 401 Homo sapiens proteins extracted from Protein Data Bank (PDB). A simple Chi-square test was used to assess the significance of deviation of the observed and expected frequencies of 59 codons at the level of individual synonymous families in the four different protein secondary structural classes. It was observed that synonymous codon families show non-randomness in codon usage in four different secondary structural classes. However,when the genes were classified according to their GC3 levels there was an increase in non-randomness in high GC3 group of genes. The non-randomness in codon usage was further tested among the same protein secondary structures belonging to four different protein folding classes of high GC3 group of genes. The results show that in each of the protein secondary structural unit there exist some synonymous family that shows class specific codon-usage pattern. Moreover, there is an increased non-random behaviour of synonymous codons in sheet structure of all secondary structural classes in high GC3 group of genes. Biological implications of these results have been discussed. PMID- 17914238 TI - Cytoview: development of a cell modelling framework. AB - The biological cell, a natural self-contained unit of prime biological importance, is an enormously complex machine that can be understood at many levels. A higher-level perspective of the entire cell requires integration of various features into coherent, biologically meaningful descriptions. There are some efforts to model cells based on their genome, proteome or metabolome descriptions. However, there are no established methods as yet to describe cell morphologies, capture similarities and differences between different cells or between healthy and disease states. Here we report a framework to model various aspects of a cell and integrate knowledge encoded at different levels of abstraction, with cell morphologies at one end to atomic structures at the other. The different issues that have been addressed are ontologies, feature description and model building. The framework describes dotted representations and tree data structures to integrate diverse pieces of data and parametric models enabling size, shape and location descriptions. The framework serves as a first step in integrating different levels of data available for a biological cell and has the potential to lead to development of computational models in our pursuit to model cell structure and function, from which several applications can flow out. PMID- 17914239 TI - Subclassification and targeted characterization of prophage-encoded two-component cell lysis cassette. AB - Bacteriophage induced lysis of host bacterial cell is mediated by a two component cell lysis cassette comprised of holin and lysozyme. Prophages are integrated forms of bacteriophages in bacterial genomes providing a repertoire for bacterial evolution. Analysis using the prophage database (http://bicmku.in:8082) constructed by us showed 47 prophages were associated with putative two component cell lysis genes. These proteins cluster into four different subgroups. In this process, a putative holin (essd) and endolysin (ybcS), encoded by the defective lambdoid prophage DLP12 was found to be similar to two component cell lysis genes in functional bacteriophages like p21 and P1. The holin essd was found to have a characteristic dual start motif with two transmembrane regions and C-terminal charged residues as in class II holins. Expression of a fusion construct of essd in Escherichia coli showed slow growth. However, under appropriate conditions, this protein could be over expressed and purified for structure function studies. The second component of the cell lysis cassette, ybcS, was found to have an N terminal SAR (Signal Arrest Release) transmembrane domain. The construct of ybcS has been over expressed in E.coli and the purified protein was functional, exhibiting lytic activity against E.coli and Salmonella typhi cell wall substrate. Such targeted sequence- structure-function characterization of proteins encoded by cryptic prophages will help understand the contribution of prophage proteins to bacterial evolution. PMID- 17914240 TI - The p53-MDM2 network: from oscillations to apoptosis. AB - The p53 protein is well-known for its tumour suppressor function. The p53-MDM2 negative feedback loop constitutes the core module of a network of regulatory interactions activated under cellular stress. In normal cells, the level of p53 proteins is kept low by MDM2, i.e. MDM2 negatively regulates the activity of p53. In the case of DNA damage, the p53-mediated pathways are activated leading to cell cycle arrest and repair of the DNA. If repair is not possible due to excessive damage, the p53-mediated apoptotic pathway is activated bringing about cell death. In this paper, we give an overview of our studies on the p53-MDM2 module and the associated pathways from a systems biology perspective. We discuss a number of key predictions, related to some specific aspects of cell cycle arrest and cell death, which could be tested in experiments. PMID- 17914241 TI - Type 2 diabetes mellitus: phylogenetic motifs for predicting protein functional sites. AB - Diabetes mellitus, commonly referred to as diabetes, is a medical condition associated with abnormally high levels of glucose (or sugar) in the blood. Keeping this view, we demonstrate the phylogenetic motifs (PMs) identification in type 2 diabetes mellitus very likely corresponding to protein functional sites. In this article, we have identified PMs for all the candidate genes for type 2 diabetes mellitus. Glycine 310 remains conserved for glucokinase and potassium channel KCNJ11. Isoleucine 137 was conserved for insulin receptor and regulatory subunit of a phosphorylating enzyme. Whereas residues valine, leucine, methionine were highly conserved for insulin receptor. Occurrence of proline was very high for calpain 10 gene and glucose transporter. PMID- 17914242 TI - The next step in biology: a periodic table? AB - Systems biology is an approach to explain the behaviour of a system in relation to its individual components. Synthetic biology uses key hierarchical and modular concepts of systems biology to engineer novel biological systems. In my opinion the next step in biology is to use molecule-to-phenotype data using these approaches and integrate them in the form a periodic table. A periodic table in biology would provide chassis to classify, systematize and compare diversity of component properties vis-a-vis system behaviour. Using periodic table it could be possible to compute higher- level interactions from component properties. This paper examines the concept of building a bio-periodic table using protein fold as the fundamental unit. PMID- 17914243 TI - Modularized study of human calcium signalling pathway. AB - Signalling pathways are complex biochemical networks responsible for regulation of numerous cellular functions. These networks function by serial and successive interactions among a large number of vital biomolecules and chemical compounds. For deciphering and analysing the underlying mechanism of such networks,a modularized study is quite helpful. Here we propose an algorithm for modularization of calcium signalling pathway of H. sapiens . The idea that "a node whose function is dependent on maximum number of other nodes tends to be the center of a sub network" is used to divide a large signalling network into smaller sub networks. Inclusion of node(s) into sub networks(s) is dependent on the outdegree of the node(s). Here outdegree of a node refers to the number of relations of the considered node lying outside the constructed sub network. Node(s) having more than c relations lying outside the expanding sub network have to be excluded from it. Here c is a specified variable based on user preference, which is finally fixed during adjustments of created sub networks, so that certain biological significance can be conferred on them. PMID- 17914244 TI - Gene ordering in partitive clustering using microarray expressions. AB - A central step in the analysis of gene expression data is the identification of groups of genes that exhibit similar expression patterns. Clustering and ordering the genes using gene expression data into homogeneous groups was shown to be useful in functional annotation, tissue classification, regulatory motif identification, and other applications. Although there is a rich literature on gene ordering in hierarchical clustering framework for gene expression analysis, there is no work addressing and evaluating the importance of gene ordering in partitive clustering framework, to the best knowledge of the authors. Outside the framework of hierarchical clustering, different gene ordering algorithms are applied on the whole data set, and the domain of partitive clustering is still unexplored with gene ordering approaches. A new hybrid method is proposed for ordering genes in each of the clusters obtained from partitive clustering solution, using microarray gene expressions.Two existing algorithms for optimally ordering cities in travelling salesman problem (TSP), namely, FRAG_GALK and Concorde, are hybridized individually with self organizing MAP to show the importance of gene ordering in partitive clustering framework. We validated our hybrid approach using yeast and fibroblast data and showed that our approach improves the result quality of partitive clustering solution, by identifying subclusters within big clusters, grouping functionally correlated genes within clusters, minimization of summation of gene expression distances, and the maximization of biological gene ordering using MIPS categorization. Moreover, the new hybrid approach, finds comparable or sometimes superior biological gene order in less computation time than those obtained by optimal leaf ordering in hierarchical clustering solution. PMID- 17914245 TI - Analysis of breast cancer progression using principal component analysis and clustering. AB - We develop a new technique to analyse microarray data which uses a combination of principal components analysis and consensus ensemble k-clustering to find robust clusters and gene markers in the data. We apply our method to a public microarray breast cancer dataset which has expression levels of genes in normal samples as well as in three pathological stages of disease; namely, atypical ductal hyperplasia or ADH, ductal carcinoma in situ or DCIS and invasive ductal carcinoma or IDC. Our method averages over clustering techniques and data perturbation to find stable, robust clusters and gene markers. We identify the clusters and their pathways with distinct subtypes of breast cancer (Luminal,Basal and Her2+). We confirm that the cancer phenotype develops early (in early hyperplasia or ADH stage) and find from our analysis that each subtype progresses from ADH to DCIS to IDC along its own specific pathway, as if each was a distinct disease. PMID- 17914246 TI - Kinetics of granulocyte colony-stimulating factor in the human milk of a nursing donor receiving treatment for mobilization of the peripheral blood stem cells. PMID- 17914247 TI - Insulin resistance, oxidative stress, and podocyte injury: role of rosuvastatin modulation of filtration barrier injury. AB - BACKGROUND/AIM: There is an emerging relationship between insulin resistance/hyperinsulinemia, oxidative stress, and glomerular injury manifesting as albuminuria. HMG-CoA reductase inhibitors (statins) have been shown to reduce oxidative stress in the vasculature as well as albuminuria in animal models and in human studies. The glomerular filtration barrier is emerging as a critical determinant of albumin filtration. We investigated the effects of insulin resistance and rosuvastatin or placebo on the glomerular filtration barrier. METHOD: Young Zucker obese and Zucker lean rats (6-7 weeks old) were treated with the HMG-CoA reductase inhibitor rosuvastatin (10 mg/kg/day) or placebo for 21 days. RESULTS: In the Zucker obese rats, homeostasis model assessment-insulin resistance index, oxidative markers (NADPH oxidase activity, reactive oxygen species, and urine isoprostane formation), podocyte foot process effacement, and albuminuria were increased as compared with Zucker lean controls, independent of increases in systolic blood pressure. Albuminuria correlated with podocyte foot process effacement (r(2) = 0.61) and insulin level (r(2) = 0.69). Rosuvastatin treatment improved albuminuria, filtration barrier indices, and oxidative stress via copper/zinc superoxide dismutase. CONCLUSIONS: These data indicate that hyperinsulinemia together with insulin resistance is associated with podocyte injury and albuminuria independent of the systolic blood pressure. Further, rosuvastatin modulates filtration barrier injury and albuminuria and improves oxidative stress measures via copper/zinc superoxide dismutase. PMID- 17914248 TI - Viability and functionality of fresh and cryopreserved human hyperplastic parathyroid tissue tested in vitro. AB - BACKGROUND/AIMS: This study aimed to test the viability and functionality of fresh and cryopreserved human hyperplastic parathyroid glands cultured in vitro. METHODS: Small fragments of 18 parathyroid glands from 18 patients with secondary hyperparathyroidism were cultured in vitro, freshly or after cryopreservation, during 60 h. Cell viability and functionality of the parathyroid fragments exposed to calcium and calcitriol were studied. RESULTS: Human parathyroid glands obtained from renal patients with secondary hyperparathyroidism maintained their viability and functionality for 60 h in culture. Sixty percent of the fresh but only 10% of the cryopreserved parathyroid glands showed the expected response with higher intact parathyroid hormone secretion when cultured with 0.6 mM calcium compared to 1.2 mM calcium. On the contrary, 44 of fresh and 40% of cryopreserved glands behaved in the same manner, showing a similar decrease in intact parathyroid hormone synthesis and secretion when cultured with calcitriol (10(-8)M). CONCLUSION: These results demonstrate differences in the response to calcium between fresh and cryopreserved glands and no differences in the response to calcitriol. This in vitro culture method may be useful to discriminate between responsive and nonresponsive hyperplastic human parathyroid glands. PMID- 17914249 TI - Urinary cytokines and steroid responsiveness in idiopathic nephrotic syndrome of childhood. AB - BACKGROUND/AIM: Steroid-resistant nephrotic syndrome (SRNS) has been associated with activation of TGF-beta(1) and progression to chronic kidney disease. Steroid sensitive nephrotic syndrome (SSNS) has been associated with activation of T cells and favorable outcome. Our objective was to distinguish SRNS from SSNS and focal segmental glomerulosclerosis (FSGS) from minimal change disease (MCD) on the basis of urinary cytokine profile. METHOD: We used a high-throughput cytokine array. ICAM-1 and TGF-beta(1) in urine and kidney tissue were evaluated by ELISA and immunohistochemistry (IHC), respectively. RESULTS: Age, gender, race, body mass index, and glomerular filtration rate were similar among groups. There were no statistically significant differences between SRNS (n = 12) and SSNS (n = 12) in regard to the presence of hypertension, treatment with ACE inhibitors, and renal histology. Arrays detected a 1- to 5.5-fold increase in urinary cytokine expression in subjects with idiopathic nephrotic syndrome (INS) as compared to controls. Using ELISA, urinary excretion of ICAM-1 was significantly higher in INS subjects than in controls (control group, n = 12; p = 0.005), but it did not differentiate SRNS from SSNS, or FSGS from MCD. IHC failed to reveal differences in renal tissue expression of ICAM-1 among controls, SRNS and SSNS. There were no significant differences among controls, and patients with SRNS and SSNS in the urinary excretion of TGF-beta(1) (p = 0.21). However, urinary TGF-beta(1) levels were significantly higher in FSGS than in MCD (p = 0.03), and IHC showed increased immunoreactivity in FSGS. CONCLUSION: Our data indicate that urinary TGF-beta(1) was able to differentiate between FSGS and MCD but was not a biomarker of steroid responsiveness. PMID- 17914250 TI - Oxidative stress and asymmetric dimethylarginine is independently associated with carotid intima media thickness in peritoneal dialysis patients. AB - BACKGROUND: Oxidative stress (OS) and asymmetric dimethylarginine (ADMA) are accepted as nonclassical cardiovascular risk factors in end-stage renal disease patients. To clarify the role of these factors in the atherosclerotic process, we investigated if OS and ADMA are associated with common carotid artery intima media thickness (CIMT) in peritoneal dialysis (PD) patients. METHODS: Thirty PD patients without known atherosclerotic disease and classical cardiovascular risk factors as well as age- and gender-matched 30 healthy individuals were included. We measured serum thiobarbituric acid-reactive substances (TBARS), malondialdehyde (MDA), advanced glycation end product (AGE), pentosidine, advanced oxidation protein products (AOPP), ADMA and CIMT in each subjects. RESULTS: TBARS, MDA, AOPP, AGE, pentosidine and ADMA levels were significantly higher in PD patients than in controls (p < 0.001). CIMT in patients was higher than in the control group (0.83 +/- 0.09 vs. 0.77 +/- 0.06 mm; p < 0.01). CIMT was independently correlated with TBARS (beta = 0.33, p < 0.01), MDA (beta = 0.27, p < 0.01), AOPP (beta = 0.22, p < 0.02), AGE (beta = 0.45, p < 0.01), pentosidine (beta = 0.56, p < 0.01) and ADMA (beta = 0.54, p < 0.01). CONCLUSIONS: OS markers and serum ADMA levels independently predict the CIMT level in PD patients. PMID- 17914251 TI - Oral paricalcitol for the treatment of secondary hyperparathyroidism in patients on hemodialysis or peritoneal dialysis. AB - BACKGROUND/AIMS: Secondary hyperparathyroidism is a common complication of chronic kidney disease, resulting from inactivation of vitamin D receptor signaling and phosphate retention. Selective activation of vitamin D receptors with intravenous paricalcitol significantly reduced parathyroid hormone (PTH) levels with no significant hypercalcemia or hyperphosphatemia in predialysis and hemodialysis (HD) patients. This study investigates the effects of oral paricalcitol to reduce PTH in patients receiving chronic HD and peritoneal dialysis (PD). METHODS: Eighty-eight patients were randomized in double-blind fashion to receive paricalcitol or placebo for 12 weeks. The dose of the study drug was adjusted weekly using the previous week's intact PTH (iPTH) level as well as calcium and Ca x P product levels. The primary end points were efficacy (two consecutive iPTH decreases of >or=30%) and safety (two consecutive calcium measurements >11.0 mg/dl). Markers of biochemical bone activity were followed. RESULTS: Demographic characteristics were similar between treatment groups. The mean paricalcitol doses (three times a week) over the entire treatment period for subjects with baseline iPTH 500 pg/ml were 3.9 and 7.6 microg, respectively. A statistically significant decrease in iPTH was seen after week 1, with a mean 30% reduction occurring by week 3. A significantly greater proportion of both HD and PD paricalcitol subjects [83% (33/40) and 100% (18/18), respectively] achieved two consecutive >or=30% decreases in iPTH. The treatment groups were not statistically different in regard to the hypercalcemia safety end point. Phosphate binder use and mean serum phosphorus levels were not different between the treatment groups. The markers of bone activity improved in the treated subjects and worsened in those on placebo. CONCLUSION: Paricalcitol provides a rapid and sustained reduction of PTH in both HD and PD patients with minimal effect on serum calcium and phosphorus and no significant difference in adverse events as compared with placebo. PMID- 17914252 TI - Preface: plasticity and diversity in behavior and brain function--important raw material for natural selection? PMID- 17914253 TI - Individual variation in coping with stress: a multidimensional approach of ultimate and proximate mechanisms. AB - Ecological studies on feral populations of mice, fish and birds elucidate the functional significance of phenotypes that differ individually in their behavioral and neuroendocrine response to environmental challenge. Within a species, the capacity to cope with environmental challenges largely determines individual survival in the natural habitat. Recent studies indicate that individual variation within a species may buffer the species for strong fluctuations in the natural habitat. A conceptual framework will be presented that is based on the view that individual variation in aggressive behavior can be considered more generally as a variation in actively coping with environmental challenges. Highly aggressive individuals adopt a proactive coping style whereas low levels of aggression indicate a more passive or reactive style of coping. Coping styles have now been identified in a range of species and can be considered as trait characteristics that are stable over time and across situations. The dimension of coping style seems to be independent of an emotionality dimension. Hence, in the analysis of the proximate mechanisms of stress and adaptation, one has to consider the possibility that the mechanisms which determine the type of stress response might be independent from those underlying the magnitude of the response. The two coping styles differ in a number of important neurobiological and neuroendocrine systems. For example, proactive males differ significantly from reactive males in the homeostatic control of serotonergic activity resulting in completely opposite dose response relationships of various serotonergic drugs. The results so far show that proactive coping is characterized by a strong inhibitory control of the 5-HT neuron via its somatodendritic 5-HT(1A) autoreceptor. It is hypothesized that the regulation of serotonin release is causally related to coping style rather than emotionality. Understanding the functional individual variation as it occurs in nature and the underlying neurobiology and neuroendocrinology is fundamental in understanding individual vulnerability to stress related disease. PMID- 17914254 TI - Genetically determined variation in stress responsiveness in rainbow trout: behavior and neurobiology. AB - It is becoming increasingly recognized that the diversity in stressors, their intensity, predictability and the context in which they are experienced, will result in behavioral and physiological responses just as diverse. In addition, stress responses are characterized by individual variations where the physiological and behavioral reactions are associated in such a manner that distinct stress coping styles encompassing suites of correlated traits can be identified. These are often referred to as proactive and reactive stress coping styles. Proactive coping is characterized by more aggression, higher general activity and higher sympathetic activation, whereas reactive coping is characterized by immobility, lack of initiative and a higher parasympathetic/hypothalamic activation. Stable coping styles appear to coexist within populations, and these strategies appear to be largely innate. Moreover, the physiological and behavioral traits of coping styles appear to be heritable. These stress coping styles have proven to play a major role in competitive ability and subsequent social position in different species of vertebrates. However, there are also studies showing that social position can affect parameters encompassing the stress coping style of individuals. In this regard it is important, but not always easy, to distinguish between causes and effects of behavioral and physiological responses to stressors. The question raised is to what extent and rigidness stress coping styles are guided by genetic factors. PMID- 17914255 TI - Social regulation of neurogenesis in teleosts. AB - Salmonid fishes such as the rainbow trout (Oncorhynchus mykiss) are frequently used to study behavioral and neuroendocrine effects of socially induced stress. A predictable aggressive response to territorial intrusion, a well described neuroanatomy, and many essential similarities in the stress response in fishes and other vertebrates are among the advantages of this comparative model. One conspicuous difference when compared to mammals, however, is that in teleost fish and other non-mammalian vertebrates, neurogenesis persists into adulthood to a much higher degree. Very little is known about the functional significance of individual differences in the rate of brain cell proliferation in fish, or whether structural changes in the fish brain are influenced by the social environment. In this paper we discuss the observation that brain cell proliferation is reduced in subordinate fish, focusing in particular on whether such individual variation reflects a difference in coping style or is indeed a response to social interactions. PMID- 17914256 TI - Genomic responses to behavioral interactions in an African cichlid fish: mechanisms and evolutionary implications. AB - Phenotypic plasticity in Astatotilapia burtoni allows individual males to alternate between dominant and subordinate status, two physiologically and behaviorally distinct phenotypes. Because these phenotypes are completely reversible, they provide an excellent model for studying the molecular mechanisms of phenotypic plasticity. The ability to express alternate phenotypes in A. burtoni depends on the ability to regulate gene expression on both short- and long-term time scales. Previous studies have demonstrated that dominant males, who have increased reproductive capacity, have higher expression of several genes involved in reproduction (e.g., genes for steroid receptors). These differences in gene expression and reproductive physiology are controlled by interactions among males. Recently, it was found that the same interactions that lead to stable long-term changes in gene expression also induce short-term and transient changes in expression of egr-1, an immediate-early gene transcription factor. This immediate-early gene response is part of a general mechanism for mediating changes in gene expression that underlie phenotypic plasticity. Longer stable changes in gene expression must involve other mechanisms, such as dynamic modifications of the epigenome. Recent data suggests a direct link between the immediate-early gene response and epigenetic modifications. These mechanisms which link behavioral interactions to changes in gene expression allow phenotypic variation to occur without corresponding changes in the genome and, as a consequence, they have implications for evolution. In the case of A. burtoni, phenotypic plasticity is likely to slow evolution because it produces highly adapted phenotypes under the primary niches encountered in the life-history of the species and the plasticity itself is likely to be an adaptive trait. PMID- 17914257 TI - Behavioral diversity and neurochemical plasticity: selection of stress coping strategies that define social status. AB - Social interactions include a variety of stimulating but challenging factors that are the basis for strategies that allow individuals to cope with novel or familiar stressful situations. Evolutionarily conserved strategies have been identified that reflect specific behavioral and physiological identities. In this review we discuss a unique model for social stress in the lizard Anolis carolinensis, which has characteristics amenable to an investigation of individual differences in behavioral responses via central and sympathetic neurochemical adaptation. Profiles of proactive and reactive phenotypes of male A. carolinensis are relatively stable, yet retain limited flexibility that allows for the development of the social system over time. For male A. carolinensis, the celerity of social signal expression and response translate into future social standing. In addition, proactive aggressive, courtship, and feeding behaviors also predict social rank, but are not as important as prior interactions and memories of previous opponents to modify behavioral output and affect social status. The central neurotransmitters dopamine and serotonin, and the endocrine stress axis (HPA) appear to be the fundamental link to adaptive stress coping strategies during social interactions. Only small adaptations to these neural and endocrine systems are necessary to produce the variability measured in behavioral responses to stressful social interactions. These neuroendocrine factors are also manifest in responses to other stimuli and form the basis of heritable strategies for coping with stress. PMID- 17914258 TI - Stress, aggression, and puberty: neuroendocrine correlates of the development of agonistic behavior in golden hamsters. AB - During puberty, agonistic behaviors undergo significant transitions. In golden hamsters, puberty is marked by a transition from play fighting to adult aggression. During early puberty, male golden hamsters perform play-fighting attacks. This response type is gradually replaced by adult attacks over the course of puberty. Interestingly, this behavioral transition does not appear to be controlled by changes in gonadal steroids. Instead, the shift from play fighting to adult aggression in male golden hamsters is driven by pubertal changes in glucocorticoid levels. Specifically, the transition from play fighting to adult aggression coincides with developmental increases in glucocorticoid levels, and external manipulations such as social stress or treatment with corticosteroid receptor agonists accelerate this behavioral shift. Moreover, the consequences of social stress differ greatly between juvenile and adult male golden hamsters. Although a single defeat during adulthood causes severe and long lasting behavioral and neuroendocrine consequences, socially subjugated juveniles show only transient behavioral effects. As such, it is likely that pubertal changes in the HPA axis are not only linked to the maturation of offensive responses but also determine the consequences of social stress. Combined, these studies in golden hamsters provide a novel mechanism for the development of agonistic behavior and suggest that age related differences in behavioral plasticity are mediated by the development of the HPA axis. PMID- 17914259 TI - Neurobiological mechanisms of aggression and stress coping: a comparative study in mouse and rat selection lines. AB - Aggression causes major health and social problems and constitutes a central problem in several psychiatric disorders. There is a close relationship between the display of aggression and stress coping strategies. In order to gain more insight into biochemical pathways associated with aggression and stress coping, we assessed behavioral and neurobiological responses in two genetically selected rodent models, namely wild house mice selectively bred for a short (SAL) and long (LAL) attack latency and Wistar rats bred for high (HAB) or low (LAB) anxiety related behavior. Compared to their line counterparts, the SAL mice and the LAB rats display a high level of intermale aggression associated with a proactive coping style. Both the SAL mice and the LAB rats show a reduced hypothalamic pituitary-adrenal (HPA) axis response to non-social stressors. However, when exposed to social stressors (resident-intruder, sensory contact), SAL mice show an attenuated HPA response, whereas LAB rats show an elevated HPA response. In both rodent lines, the display of aggression is associated with high neuronal activation in the central amygdala, but reduced neuronal activation in the lateral septum. Furthermore, in the lateral septum, SAL mice have a reduced vasopressinergic fiber network, and LAB rats show a decreased vasopressin release during the display of aggression. Moreover, the two lines show several indications of an increased serotonergic neurotransmission. The relevance of these findings in relation to high aggression and stress coping is discussed. In conclusion, exploring neurobiological systems in animals sharing relevant behavioral characteristics might be a useful approach to identify general mechanisms of action, which in turn can improve our understanding of specific behavioral symptoms in human psychiatric disorders. PMID- 17914260 TI - Association between vitamin D receptor gene polymorphisms and susceptibility to chronic kidney disease and periodontitis. AB - BACKGROUND/AIMS: Chronic kidney disease (CKD) and periodontitis (PD) are serious public-health concerns. Vitamin D is a fat-soluble steroid hormone that interacts with its nuclear receptor (VDR) to regulate a variety of biological processes, such as bone metabolism, immune response modulation and transcription of several genes involved in CKD and PD disease mechanisms. The aim of this work was to investigate the association between polymorphisms in the VDR gene and end-stage renal disease (ESRD) and PD. METHODS: 222 subjects with and without ESRD (in hemodialysis) were divided into groups with and without PD. Polymorphisms TaqI and BsmI in the VDR gene were analyzed by PCR restriction fragment length polymorphism. The significance of differences in allele, genotype and haplotype frequencies between groups was assessed by the chi2 test (p value <0.05) and odds ratio (OR). RESULTS: Allele G was associated with protection against ESRD: groups without versus with ESRD (GG) x (GA+AA): OR = 2.5, 95% CI = 1.4-4.6, p = 0.00; (G x A): OR = 1.5, 95% CI = 1.0-2.3, p = 0.02; (TG + CG) x (TA + CA): OR = 1.5, 95% CI = 1.0-2.3, p = 0.02. No association was observed between the study polymorphisms and susceptibility to or protection against PD. CONCLUSION: Allele G of the VDR BsmI polymorphism was associated with protection against ESRD. PMID- 17914261 TI - Increase in beta-amyloid levels in cerebrospinal fluid of children with Down syndrome. AB - BACKGROUND: Individuals with Down syndrome (DS) invariably develop Alzheimer's disease (AD) during their life span. It is therefore of importance to study young DS patients when trying to elucidate early events in AD pathogenesis. AIM: To investigate how levels of different amyloid-beta (Abeta) peptides, as well as tau and phosphorylated tau, in cerebrospinal fluid (CSF) from children with DS change over time. The first CSF sample was taken at 8 months and the following two samples at 20-40 and 54 months of age. RESULTS: Individual levels of the Abeta peptides, as well as total Abeta levels in CSF increased over time when measured with Western blot. Tau in CSF decreased whereas there was no change in levels of phosphorylated tau over time. CONCLUSION: The increasing levels of Abeta in CSF during early childhood of DS patients observed in this study are probably due to the trisomy of the Abeta precursor APP, which leads to an overproduction of Abeta. Despite the increased CSF concentrations of Abeta, there were no signs of an AD-indicating tau pattern in CSF, since the levels of total tau decreased and phosphorylated tau remained unchanged. This observation further strengthens the theory of Abeta pathology preceding tau pathology in AD. PMID- 17914262 TI - Is functional decline necessary for a diagnosis of Alzheimer's disease? AB - BACKGROUND: The purpose of this study is to examine baseline differences and annualized cognitive and functional change scores in mild Alzheimer's disease (AD) patients with and without impaired activities of daily living (ADL). METHODS: We recruited 267 mild probable AD patients with at least 1 year of follow-up (NINCDS-ADRDA criteria, MMSE>or=20). Based on initial ADL scores, they were divided into 2 groups: unimpaired (n=40) and impaired (n=227). We compared the differences in annualized change scores on MMSE, Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-cog), ADL and Clinical Dementia Rating sum of box score (CDR-SB) for patients with and without functional impairment at baseline. RESULTS: The group with unimpaired ADL at baseline had a significantly shorter symptom duration (p=0.01) and better neuropsychological test scores at baseline (p<0.001) than those with impaired ADL. The annualized cognitive and functional change of each group from baseline to 1-year follow-up was not significantly different on the MMSE, ADAS-cog, CDR-SB, Physical Self-Maintenance Scale and Instrumental Activities of Daily Living. After 1 year, 56% of the initially unimpaired group and 6% of the initially impaired group reported no ADL impairment. CONCLUSIONS: Our study suggests that functional decline should not be required for the diagnosis of mild AD. PMID- 17914263 TI - [Hypertensive diseases during pregnancy: an interdisciplinary challenge]. AB - Hypertensive diseases in pregnancy are one of the main reasons of maternal, fetal and neonatal morbidity and mortality. As preeclampsia is a multisystem disorder with different clinical characteristics, prevention, diagnosis and therapy of this disease require a close interdisciplinary cooperation. The present papers will summarize the status quo of the management of preeclampsia. PMID- 17914264 TI - [Pathophysiology of preeclampsia]. AB - Preeclampsia is a multisystem disorder affecting about 5-10% of all pregnancies. It is a major cause of maternal, fetal and neonatal mortality and morbidity. Despite intensive research the etiology of this disease still remains unknown. Until now the inadequate transformation of the smooth-muscle cells of spiral arteries in the placental bed caused by an insufficient endovascular invasion of the trophoblast has been considered to be the major reason for the development of preeclampsia. Maternal-fetal (paternal) immune maladaptation is implicated in the insufficient trophoblast invasion, which leads to an imbalance of angiogenic and antiangiogenic factors at the maternal-fetal interface. This review summarizes the actual knowledge of important pathophysiological basic principles of preeclampsia. PMID- 17914265 TI - [Prophylactic treatment of preeclampsia]. AB - The pathophysiology of preeclampsia is still unknown. Low-dose aspirin showed a reduction of the preeclampsia rate in a high-risk situation.There is a lack of prospective randomized studies to prove the benefit of heparin treatment in patients with hereditary thrombophilia. Different prophylactic therapies did not show any benefit in the prevention of preeclampsia in clinical studies. PMID- 17914266 TI - [Current recommendations for the treatment of preeclampsia]. AB - Low-dose aspirin, but not vitamin C and vitamin E, has small to moderate benefits when used for the prevention of preeclampsia. There is however little to suggest that the course of manifest preeclampsia can be substantially affected by drug treatment. Antihypertensive therapy increases the risk of fetal growth restriction and is only indicated in severe hypertension to reduce the risk of maternal cerebral hemorrhage. Magnesium sulfate is the drug of choice for the prevention and treatment of eclampsia. Volume expansion as well as the use of steroids in preeclamptic patients without HELLP syndrome have failed to show any improvement. Delivery as the only definitive treatment is always beneficial for the mother whereas expectant management is in favor of a premature fetus. Decision-making largely depends on gestational age and severity of the disease. PMID- 17914267 TI - [Current management of the HELLP syndrome]. AB - The HELLP syndrome as part of the microangiopathic syndromes requires special attention in terms of a rapid and accurate diagnostic and differential diagnostic workup because of its possibly rapid clinical deterioration. It is defined by the classical triad of hemolysis,elevated liver enzymes and low platelet counts which may lead to prognostically relevant problems in differentiating it from thrombotic-thrombocytopenic purpura and hemolytic-uremic syndrome and other pregnancy-related and unrelated liver diseases, i.e. mainly clinical and laboratory similarities to other liver diseases such as acute fatty liver or intrahepatic cholestasis in pregnancy or pregnancy-unrelated settings like viral hepatitides. The management in the different phases of pregnancy is described in detail. Therapeutic options to prolong pregnancy are discussed as are the possibilities of prophylaxis in subsequent pregnancies and aspects of the followup. PMID- 17914268 TI - [The 'Arbeitsgemeinschaft Gestose-Frauen e.V.': A self-help organization with extensive experience in the aftercare of women with preeclampsia or HELLP syndrome]. AB - The 'Arbeitsgemeinschaft Gestose-Frauen e.V.' was founded in 1984 with a couple of sufferers and has advised more than 24,000 women so far. In addition to the care by the midwife and gynaecologist, this self-help organization offers the possibility to exchange experiences with other sufferers, but also to receive medical and psychological counselling. PMID- 17914270 TI - [Ogilvie's syndrome (acute idiopathic colonic pseudo-obstruction) after Caesarean section]. AB - Due to incipient HELLP syndrome and no progression in labour, a caesarean was performed on a 33-year-old para I gravida I. During the operation she suffered a heavy blood loss as a result of uterine inertia. In the postoperative stage she developed an acute idiopathic colonic pseudo-obstruction, also known as Ogilvie's syndrome. Only 6 days after the operation did she slowly recover. Additional factors that could have led to Ogilvie's syndrome are the caesarean, the HELLP syndrome and the postpartal haemorrhage. Due to her reduced physical health, the mobilization of the patient was restricted, which further prolonged the recovery. However, an early initiation of supportive treatment, the timely diagnosis of Ogilvie's syndrome and two colonoscopic decompressions prevented a caecal rupture. After 3 weeks of hospitalization, the patient was discharged in good general health. PMID- 17914269 TI - [Diagnosis of vulval and vaginal neoplasms with magnetic resonance imaging]. AB - Magnetic resonance imaging (MRI) is widely accepted as an established method in the diagnostic imaging of diseases of the female pelvis. This review article describes the role of MRI in the diagnosis of neoplasms of the vulva and the vagina. MRI is useful in the pretherapeutic staging of vulval and vaginal neoplasms in order to assess tumor size, local tumor extent and presence of lymph node metastases. Further indications for MRI are the diagnosis of posttherapeutic changes and tumor recurrence. PMID- 17914271 TI - Advancing women's cancer care: highlights from the Annual Meeting on Women's Cancer 2007. Report on the 38th Annual Meeting of the Society of Gynecologic Oncologists, San Diego, Calif., March 3-7, 2007. PMID- 17914272 TI - Enzymatic activity in body and fecal extracts of the storage mite Chortoglyphus arcuatus. AB - BACKGROUND: Chortoglyphus arcuatus has been described in many countries. Many allergens are potent enzymes, which may promote a Th2 immune response. The aim of this study was to evaluate the enzymatic activity of body and fecal extracts of C. arcuatus. MATERIAL AND METHODS: Feces and bodies of full-grown C. arcuatus cultures were separated by sieving, extracted in PBS, dialyzed and lyophilized. The antigenic profile of both extracts was determined by SDS-PAGE. Immunoblot experiments were conducted using a pool of sera from allergic individuals residing in Galicia, a region of Spain, where this species is abundant. The enzymatic activity of the extracts was evaluated by the zymogram technique. Serine and cysteine protease activity was measured using in vitro methods. The API Zym system was used to determine the enzymatic properties of the extracts. RESULTS: The antigenic profile showed that the body extract contained more and better defined bands than the fecal extract. Allergens were detected in both extracts in a molecular weight range between 14 and 100 kDa. Gelatinolytic gels confirmed that fecal extracts contain more hydrolytic enzymatic activity than body extracts. Serine protease activity in fecal extracts was higher than in body extracts (5.98 vs. 2.701 IU of trypsin/mg of freeze-dried material). No cysteine protease activity was detected. CONCLUSION: C. arcuatus extracts contain several allergens and proteins with high enzymatic activity, especially in the feces. Some of these allergens may be enzymes. Fecal extracts have more enzymatic activity than body extracts. PMID- 17914273 TI - Pollen grains induce a rapid and biphasic eczematous immune response in atopic eczema patients. AB - INTRODUCTION: Eczematous reactions to type I allergy-inducing antigens are documented in a subgroup of patients with atopic eczema. Yet, the underlying immunological mechanisms are not well understood. MATERIAL AND METHODS: To delineate the effect of native pollen grains on human skin of healthy and atopic individuals we performed patch tests (atopy patch test with native pollen grains, PPT). Nickel patch tests (NPT) served as an established model of contact dermatitis. Skin site biopsies were taken 6-96 h after allergen application and investigated immunohistochemically. RESULTS: Histology of positive patch tests showed an influx of mononuclear cells (predominantly CD4+, CD25+, CD45RO+). This influx was detected earlier in the PPT reaction than in the immune response to nickel. A biphasic cytokine response could be detected in the PPT: IL-5 dominated in the early, IFN-gamma in the late phase. The NPT was continuously dominated by IFN-gamma. Dendritic cell subpopulations imitated the earlier kinetics of the mononuclear infiltrate. DISCUSSION: Thus, pollen grains induce eczematous reactions in susceptible individuals. This reaction appears clinically and immunohistochemically similar to the contact hypersensitivity reaction to nickel but follows a faster kinetic and a biphasic course: Th2 and IgE in the early (24 h) and Th1 predominance in the late (96 h) phase. PMID- 17914274 TI - Date palm pollen allergoid: characterization of its chemical-physical and immunological properties. AB - BACKGROUND: Date palm (DP) pollen can cause allergic symptoms in people living in different countries. Specific immunotherapy with allergenic extracts by subcutaneous route is effective to cure allergic people. However, the risk of side effects has led to explore safer therapeutic modalities. The aim of our work was to evaluate IgE cross-reactivity between DP and autochthonous palm (European fan palm, EFP) pollen extracts, to chemically modify DP extract with potassium cyanate in order to obtain an allergoid, and to characterize it. METHODS: By radioallergosorbent test inhibition, immunoblotting (IB) and skin prick test, in vitro and in vivo allergenic activities of native and modified DP extracts were compared. By SDS-PAGE and IB, we compared the protein profile and IgE-binding capacity of both native and modified DP, as well as of EFP extracts. By IB inhibition, IgE cross-reactivity of native DP and EFP extracts was evaluated. By ELISA, the capacity of modified DP-induced IgG to react with native DP extract was determined. RESULTS: Radioallergosorbent test inhibition, IB and skin prick test results demonstrated that modified DP was significantly less allergenic than native DP extract. The SDS-PAGE profile showed that potassium cyanate treatment of DP extract did not alter the molecular weight of its components. In addition, no difference was observed between native DP and EFP extracts. Subsequent IB inhibition data evidenced the existence of a strong IgE cross-reactivity between native DP and EFP extracts. ELISA results indicated that the administration of modified DP in mice was able to induce specific IgG also recognizing native DP extract. CONCLUSIONS: Modified DP extract (allergoid) seems to be a good candidate for immunotherapy of patients affected by specific allergy. PMID- 17914275 TI - Accumulation of regulatory T cells in local draining lymph nodes of the lung correlates with spontaneous resolution of chronic asthma in a murine model. AB - BACKGROUND: Mice sensitized to ovalbumin develop allergic airway disease (AAD) with short-term aerosol challenge; however, airway inflammation resolves with long-term aerosol challenge, referred to as local inhalational tolerance (LIT). METHODS: We sought to determine if resolution of airway inflammation correlated with increases in lymphocyte subsets in local lung compartments, including putative regulatory T cells. RESULTS: At the AAD stage, total numbers of T and B lymphocytes in bronchoalveolar lavage (BAL) were significantly increased above controls; however, at LIT, T and B lymphocytes were significantly reduced compared to AAD. In the lung tissue, the only alteration was a significant increase in CD4+ CD25+ T cells at AAD. In the hilar lymph node (HLN), CD4+ and CD4+ CD25+ T cells were significantly increased at AAD and LIT. In addition, CD8+ T cells were significantly elevated in the HLN at LIT, and CD19+ B cells were significantly increased at AAD. Adoptive transfer of HLN lymphocytes to lymphopenic mice confirmed that AAD lymphocytes could induce airway inflammation in response to aerosol challenge, whereas LIT lymphocytes were unable to do so. Depletion of CD4+ CD25+ T cells in vivo resulted in exacerbation of inflammation at AAD and LIT. CD4+ CD25+ T cells in the HLN also displayed suppressive activity in vitro. Additionally, T cells expressing Foxp3 were increased in the BAL and HLN during LIT. CONCLUSIONS: These results indicate that lymphocytes with regulatory functions are increased and sustained in local lung compartments at LIT and that their appearance correlates with the resolution of lung inflammation. PMID- 17914276 TI - Risk factors leading to hospital admission in Iranian asthmatic children. AB - BACKGROUND: Asthma is one of the most common chronic diseases in the world, leading to an increased rate of hospitalization. We performed this study to better understand the factors leading to admission among asthmatic children. METHODS: We performed a study among asthmatic children in a referral hospital for asthma and allergy in Tehran. Sixty-three cases were selected from asthmatic children admitted to the emergency room (ER) who still had an indication for ward or intensive care unit admission after primary treatment. Our control group was the asthmatic children discharged after primary treatment and patients who were referred to the asthma and allergy clinic (63 patients). Data were obtained by structured questionnaires filled out during clinical interviews. RESULTS: There was a significant difference in mean age (5 years for cases vs. 6 years for controls; p = 0.049), personal and familial allergic history (69.8 and 57.1% for cases vs. 34.9 and 36.5% for controls; p < 0.01 and p = 0.02, respectively), history of recent respiratory infections (79.4% for cases vs. 49.2% for controls; p < 0.01), hospitalization history due to asthma (57.1% for cases vs. 23.8% for controls; p < 0.01) and regular use of inhaled corticosteroid (66.7% for cases vs. 33.3% for controls; p < 0.01). CONCLUSIONS: Our findings confirm most previous observations, suggesting that recent respiratory infections, hospitalization, personal or familial allergy, disease severity and lower ages are important factors leading to hospitalization. We also found that regular clinical follow-up, regular use of inhaled corticosteroids, higher IgE levels and O2 saturation may lower the probability of hospitalization during asthmatic attacks. PMID- 17914277 TI - Antiallergic effects of Lactobacillus pentosus strain S-PT84 mediated by modulation of Th1/Th2 immunobalance and induction of IL-10 production. AB - BACKGROUND: Many types of fermented food are consumed in Japan. Although some are produced by plant-origin lactic-acid bacteria (LAB) fermentation, the physiological functions of such bacteria remain unclear. We therefore isolated LAB of plant origin from Kyoto pickles and determined the immunological activity of heat-killed preparations of plant-origin LAB. METHODS: The Lactobacillus pentosus strain S-PT84 was selected from among 16 LAB of plant origin as the strongest interleukin (IL)-12-inducing strain. IL-12- and IL-10-inducing activities were determined with macrophages from BALB/c mice. The in vivo immunomodulating effect of S-PT84was determined with BALB/c mice fed S-PT84. The antiallergic activity of S-PT84 was examined in ovalbumin (OVA)/alum-administered BALB/c mice. RESULTS: The L. pentosus strain S-PT84 induced production of both IL 12 and IL-10 in vitro. S-PT84 enhanced splenic natural-killer activity and modulated the T helper (Th) type 1/type 2 balance toward a Th1-dominant state. In the OVA-induced allergy model, orally administered S-PT84 lowered serum IgE levels and suppressed active cutaneous anaphylaxis reaction and splenic IL-4 production. IL-10 production from splenocytes of OVA-immunized mice was upregulated by feeding S-PT84. CONCLUSIONS: Despite heat-killing, S-PT84 exhibited antiallergic effects by modulating the Th1/Th2 balance and inducing regulatory T cells. The L. pentosus strain S-PT84, which is of plant origin and isolated from a traditional Japanese food, is expected to be useful for treatment of many immune diseases including allergies, tumors, infectious diseases and auto immune diseases. PMID- 17914278 TI - Voice register in Khmu': experiments in production and perception. AB - Some Khmu' dialects have phonologically distinctive voice registers. Auditory observations have claimed a stable distinction between clear voice and high pitch for Register 1 and breathy voice and low pitch for Register 2 in the Khmu' Rawk dialect of northern Thailand. Word pairs distinguished only by register were recorded by 25 native speakers. Acoustic analysis yielded F0 and overall amplitude contours, frequencies of F1 and F2 in quasi-steady states of the vowels, relative intensities of higher harmonics to that of the first harmonic, and vowel durations. When circumstances caused early attention to perception testing, the words of only 8 speakers had been analyzed for properties other than amplitude and F0. Since the only significant factor that had emerged by then was F0 contour, the synthetic stimuli were made with just a series of seven contours. The labeling by 32 native speakers yielded two categories, demonstrating the sufficiency of F0 as an acoustic cue. The completed acoustic analysis showed a significant effect of one of the harmonic ratios for the women only, suggesting a conservative bias. The language has been shifting toward tonality and may have reached it. PMID- 17914279 TI - The 'trough effect': an ultrasound study. AB - Bilabial stops often show a lowering of the tongue in symmetrical VCV sequences. The causes of this phenomenon, sometimes called the 'trough effect', are unknown. However, it could have important implications for the study of time-varying aspects of speech events. Ultrasound is a non-invasive technique that has allowed us to image the shape of the tongue in real time and measure the actual tongue displacement that occurs in the C of a VCV sequence. Five repetitions of symmetrical V(1)CV(2) sequences with the bilabial stops /b, p/ were obtained from 10 British English speakers. Results showed not only differences in the direction and degree of the tongue displacement but also differences in the tongue contour configuration between subjects. This study demonstrates the effectiveness of ultrasound as a technique in phonetic research, making possible the analysis of tongue surface movement for large amounts of data from multiple subjects. PMID- 17914280 TI - Effects of acoustic variability in the perceptual learning of non-native-accented speech sounds. AB - This study addressed whether acoustic variability and category overlap in non native speech contribute to difficulty in its recognition, and more generally whether the benefits of exposure to acoustic variability during categorization training are stable across differences in category confusability. Three experiments considered a set of Spanish-accented English productions. The set was seen to pose learning and recognition difficulty (experiment 1) and was more variable and confusable than a parallel set of native productions (experiment 2). A training study (experiment 3) probed the relative contributions of category central tendency and variability to difficulty in vowel identification using derived inventories in which these dimensions were manipulated based on the results of experiments 1 and 2. Training and test difficulty related straightforwardly to category confusability but not to location in the vowel space. Benefits of high-variability exposure also varied across vowel categories, and seemed to be diminished for highly confusable vowels. Overall, variability was implicated in perception and learning difficulty in ways that warrant further investigation. PMID- 17914281 TI - The influence of ambient speech on adult speech productions through unintentional imitation. AB - This paper deals with the influence of ambient speech on individual speech productions. A methodological framework is defined to gather the experimental data necessary to feed computer models simulating self-organisation in phonological systems. Two experiments were carried out. Experiment 1 was run on French native speakers from two regiolects of Belgium: two from Liege and two from Brussels. When exposed to the way of speaking of the other regiolect via loudspeakers, the speakers of one regiolect produced vowels that were significantly different from their typical realisations, and significantly closer to the way of speaking specific of the other regiolect. Experiment 2 achieved a replication of the results for 8 Mons speakers hearing a Liege speaker. A significant part of the imitative effect remained up to 10 min after the end of the exposure to the other regiolect productions. As a whole, the results suggest that: (i) imitation occurs automatically and unintentionally, (ii) the modified realisations leave a memory trace, in which case the mechanism may be better defined as 'mimesis' than as 'imitation'. The potential effects of multiple imitative speech interactions on sound change are discussed in this paper, as well as the implications for a general theory of phonetic implementation and phonetic representation. PMID- 17914282 TI - The signalling of German rising-falling intonation categories--the interplay of synchronization, shape, and height. AB - Based on the phonology of the Kiel Intonation Model (KIM), a tripartite opposition of German intonation is investigated: early, medial, and late peaks. These intonation categories, which can be projected onto H + L*, H*, and L* + H in the AM framework, are described in the KIM as rising-falling F(0) peak patterns differentiated by their synchronization with the accented-vowel onset. Perception experiments were carried out, showing that the function-based identification of the peak categories is not only influenced by peak synchronization, but also by peak shape and height. While the complete spectrum of findings is not covered by the current phonological modelling, the findings corroborate the existence of all three categories in German intonation and support the idea that the timing of the peak movements with regard to the accented vowel is important for their perceptual differentiation. PMID- 17914283 TI - Causes and consequences of the DNA damage response. AB - A prerequisite for maintaining genome stability in all cell types is the accurate repair and efficient signaling of DNA double strand breaks (DSBs). It is believed that DSBs are initially detected by damage sensors that trigger the activation of transducing kinases. These transducers amplify the damage signal, which is then relayed to effector proteins, which regulate the progression of the cell cycle, DNA repair and apoptosis. Errors in the execution of the repair and/or signaling of DSBs can give rise to multi-systemic disorders characterized by tissue degeneration, infertility, immune system dysfunction, age-related pathologies and cancer. This special Spotlight issue of Cell Cycle highlights recent advances in our understanding of the biology and significance of the DNA damage response. A range of issues are addressed including mechanistic ones: What is the aberrant DNA structure that triggers the activation of the checkpoint? How does chromatin structure influence the recruitment of repair and checkpoint proteins? How does chromosomal instability contribute to the evolution of cancer? In addition, questions related to the physiology of the DNA damage response in normal and abnormal cells is explored: What is the in vivo consequence of altering specific amino acids in a DNA damage sensor? Does DNA damage accumulation in stem cells cause aging? How is neurodegeneration linked to deficiencies in specific DNA repair pathways? And finally, what is the biological basis for selection of aberrant DNA damage responses in cancer cells? PMID- 17914284 TI - c-Myc blazing a trail of death: coupling of the mitochondrial and death receptor apoptosis pathways by c-Myc. AB - TRAIL ligand induces selectively apoptosis in tumor cells by binding to two death receptors (DR4 and DR5) and holds promise as a potential therapeutic agent against cancer. While it has been known for long time that TRAIL receptors are commonly expressed in wide variety of normal tissues, it is not well understood why TRAIL kills tumor cells but leaves normal cells unharmed. The prototypic oncogene c-Myc promotes the cell cycle and simultaneously primes activation of the Bcl-2 family controlled mitochondria apoptosis pathway. A striking reflection of the c-Myc-dependent apoptotic sensitization is the dramatic c-Myc-induced vulnerability of cells to TRAIL and other death receptor ligands. Here we summarize the recent findings regarding the death mechanisms of TRAIL/TRAIL receptor system and the connection of c-Myc to the mitochondrial apoptosis pathway, focusing on our work that couples c-Myc via Bak to the TRAIL death receptor pathway. Finally, we present a mitochondria-priming model to explain how c-Myc-Bak interaction amplifies the TRAIL-induced caspase 8-Bid pathway to induce full-blown apoptosis. We discuss the implications of these findings for understanding the selective cytotoxicity of TRAIL and for the therapeutic exploitation of the death receptor pathway. PMID- 17914285 TI - Evidence-based human resources: the case for exit interview surveys. AB - This bimonthly department, sponsored by the American Organization of Nurse Executives, presents information to assist nurse leaders in shaping the future of healthcare through creative and innovative leadership. The strategic priorities of the American Organization of Nurse Executives anchor the editorial content. They reflect contemporary healthcare and nursing practice issues that challenge nurse executives as they strive to meet the needs of patients. In this article, the author discusses exit interview surveys that identify reasons why employees leave their organization, what may have encouraged them to stay, and what would entice them back to the organization. PMID- 17914286 TI - Nursing's contributions to innovative hospital design. AB - With nearly $200 billion of healthcare construction expected by 2015, nurse leaders must expand their knowledge and capabilities in healthcare design. This bimonthly department, edited by Jaynelle F. Stichler, DNSc, RN, FACHE, prepares nurse leaders to use the evidence-based design process to ensure that new, expanded, and renovated hospitals facilitate optimal patient outcomes, enhance the work environment for healthcare providers, and improve organizational performance. The guest authors of this article feature a new interdisciplinary model of graduate education that is focused on preparing design and healthcare professionals to work collaboratively in creating optimal healthcare designs that foster safety and efficiency. PMID- 17914287 TI - Clinical nurse leader and clinical nurse specialist role delineation in the acute care setting. AB - More than 90 members of the American Association of Colleges of Nursing and 190 practice sites have partnered to develop the clinical nurse leader (CNL) role. The partnership has created synergy between education and practice and nurtured innovation and diffusion of learning on a national basis. In this ongoing department, the editor, Jolene Tornabeni, MA, RN, FAAN, FACHE, showcases a variety of nurse leaders who discuss their new patient care delivery models in preparation for the CNL role and CNLs who highlight partnerships with their clinical colleagues to improve patient care. In this article, the authors explore differences and similarities between the CNL and the clinical nurse specialist roles, describing the working strategies between a CNL and clinical nurse specialist, and role delineations that have resulted from their cooperation, collaboration, and planning. PMID- 17914288 TI - Diffusing confusion among evidence-based practice, quality improvement, and research. AB - In this department, hot topics in nursing outcomes, research, and evidence-based practice relevant to the nurse administrator are highlighted. The goal is to discuss the practical implications for nurse leaders in diverse healthcare settings. Content includes evidence-based projects and decision making, locating measurement tools for quality improvement and safety projects, using outcome measures to evaluate quality, practice implications of administrative research, and exemplars of projects that demonstrate innovative approaches to organizational problems. PMID- 17914289 TI - Planning: what we can learn from complex systems science. AB - As systems evolve over time, their natural tendency is to become increasingly more complex. Studies in the field of complex systems have generated new perspectives on managing in social organizations such as hospitals. Much of this research appears as a natural extension of the cross-disciplinary field of systems theory. This is the first article in a 5-part series on applying complex systems science to the traditional management concepts of planning, organizing, directing, coordinating, and controlling. In this article, the concept of planning is explored from a complex systems' perspective. PMID- 17914290 TI - Professional nurse councils: a new model to create excitement and improve value and productivity. AB - Chief nurse executives and hospital administrators continue to debate the efficacy of empowering nurses through the structure of a nurse practice council. The author presents a new approach to a successful strategy to re-engineer an organization's nurse practice council that increases the value to the organization and creates excitement and energy in the staff with improved productivity. PMID- 17914291 TI - Transforming care at the bedside: implementation and spread model for single hospital and multihospital systems. AB - Institute of Medicine reports provide evidence of the failings of the healthcare system in the United States and a vision of the required transformation. The Institute for Healthcare Improvement and the Robert Wood Johnson Foundation created the Transforming Care at the Bedside initiative in 2003 to develop and validate a process for transforming care in hospital medical-surgical units. The authors describe Transforming Care at the Bedside as implemented by one of Institute for Healthcare Improvement/Robert Wood Johnson's initial pilot hospitals, including promising outcomes and a model for spreading the initiative. PMID- 17914292 TI - How unit level nursing responsibilities are structured in US hospitals. AB - OBJECTIVES: To describe (1) the extent to which acute and intensive care units use the elements of nursing models (team, functional, primary, total patient care, patient-focused care, case management) and (2) the deployment of non-unit based personnel resources. BACKGROUND: The lack of current data-based behavioral descriptions of the extent to which elements of nursing models are implemented makes it difficult to determine how work models may influence outcomes. METHODS: Nurse managers of 56 intensive care units and 80 acute care adult units from 40 randomly selected US hospitals participated in a structured interview regarding (1) day-shift use of patient assignment behaviors associated with nursing models and (2) the availability and consistency of assignment of non-unit-based support personnel. RESULTS: No model was implemented fully. Almost all intensive care units reported similar assignment behaviors except in the consistency of patient assignment. Non-intensive care units demonstrated wide variation in assignment patterns. Patterns differed intra-institutionally. There were large differences in the availability and deployment of non-unit-based supportive resources. CONCLUSIONS: Administrators must recognize the differences in work models within their institutions as a part of any quality improvement effort. Attempts to test new work models must be rigorous in the measurement of their implementation. PMID- 17914294 TI - Leadership character: antidote to organizational fatigue. AB - Despite the tremendous gains nurse leaders have made in shaping the environment of care, an observable, dysfunctional dynamic resulting in organizational fatigue threatens the sustainability of those gains. At its peak, this dynamic cascades through all levels of nursing leadership, causing exhaustion, diminished decision making, and reluctance by nurses to assume formal leadership roles. In the worst case, this dynamic reaches the bedside with the potential to cause harm to patients. The authors offer a prescription for organizational fatigue, based on the cross-cultural research of Dr Angeles Arrien. Following this prescription can lead to healthier leaders, stronger nursing organizations, a richer pipeline of future nurse leaders, and a safer environment for patients. PMID- 17914293 TI - Decrease new graduate nurse orientation costs by using HESI exit exam scores. AB - Predicting workplace success is becoming increasingly important because the cost of orienting new nurse graduates is increasing at such an alarming rate. This study assessed the effectiveness of the HESI Exit Exam in measuring entry-level competencies of novice nurses. Findings indicate that the HESI Exit Exam was an effective predictor of workplace competency for new graduates (N = 108) assigned to acute care and critical care units in a large, tertiary care hospital. PMID- 17914295 TI - Registered nurse job satisfaction and collective bargaining unit membership status. AB - OBJECTIVE: To examine differences in job satisfaction levels between registered nurses who were or were not members of a nursing collective bargaining unit. BACKGROUND: The nursing shortage could lead to decreased quality of patient care, heavier workloads, and mandatory overtime, resulting in decreased job satisfaction and increased intent to leave. Nursing collective bargaining units use contracts to help decrease patient workload, decrease and eliminate mandatory overtime, increase pay and benefits, and increase job security. Exploring differences in job satisfaction between nurses who are and are not members of collective bargaining units is pertinent to understanding the retention and recruitment of nurses. METHODS: A descriptive secondary analysis using a survey database from the Minnesota Department of Health. The survey, which included a job satisfaction section largely based on the Index of Work Satisfaction, was sent in 2002 to 3,645 registered nurses in Minnesota. RESULTS: Members of collective bargaining units had higher satisfaction with wages. Nonmembers had higher satisfaction with nursing supervision, patient care, work setting, professional relationships, and overall job satisfaction. CONCLUSIONS: There is a need for interventions in institutions with collective bargaining units to improve job satisfaction, nurse retention, and job recruitment. PMID- 17914296 TI - Variety is the spice of leadership. PMID- 17914297 TI - The role of a nurse retention coordinator: one perspective. PMID- 17914298 TI - Health information exchange: be knowledgeable, get involved. PMID- 17914299 TI - Making research come alive at the bedside. PMID- 17914301 TI - Positions of authority may equal lawsuits. PMID- 17914302 TI - Bugs among us. PMID- 17914304 TI - "Smart" pumps: provide safety while promoting teamwork. PMID- 17914305 TI - Improving morale through photovoice technology. PMID- 17914307 TI - Design and implementation of a portable physiologic data acquisition system. AB - OBJECTIVE: To describe and report the reliability of a portable, laptop-based, real-time, continuous physiologic data acquisition system (PDAS) that allows for synchronous recording of physiologic data, clinical events, and event markers at the bedside for physiologic research studies in the intensive care unit. DESIGN: Descriptive report of new research technology. SETTING: Adult and pediatric intensive care units in three tertiary care academic hospitals. PATIENTS: Sixty four critically ill and injured patients were studied, including 34 adult (22 males and 12 females) and 30 pediatric (19 males and 11 females). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Data transmission errors during bench and field testing were measured. The PDAS was used in three separate research studies, by multiple users, and for repeated recordings of the same set of signals at various intervals for different lengths of time. Both parametric (1 Hz) and waveform (125-500 Hz) signals were recorded and analyzed. Details of the PDAS components are explained and examples are given from the three experimental physiology-based protocols. Waveform data include electrocardiogram, respiration, systemic arterial pressure (invasive and noninvasive), oxygen saturation, central venous pressure, pulmonary arterial pressure, left and right atrial pressures, intracranial pressure, and regional cerebral blood flow. Bench and field testing of the PDAS demonstrated excellent reliability with 100% accuracy and no data transmission errors. The key feature of simultaneously capturing physiologic signal data and clinical events (e.g., changes in mechanical ventilation, drug administration, clinical condition) is emphasized. CONCLUSIONS: The PDAS provides a reliable tool to record physiologic signals and associated clinical events on a second-to-second basis and may serve as an important adjunctive research tool in designing and performing clinical physiologic studies in critical illness and injury. PMID- 17914309 TI - Congenital cystic adenomatoid malformation type 0-a rare cause of neonatal death. AB - OBJECTIVE: We give the first account of failure of extracorporeal membrane oxygenation therapy secondary to congenital cystic malformation of the lung (CCAM) type 0. DESIGN: Case report. SETTING: Pediatric intensive care unit. PATIENT: A female neonate, appropriate for gestational age, with respiratory failure immediately after delivery. INTERVENTIONS: : Cardiopulmonary support with venoarterial extracorporeal membrane oxygenation. RESULTS: There was no improvement of pulmonary function, and the patient died. CCAM type 0 was diagnosed postmortem. CONCLUSIONS: CCAM type 0 should be considered as a rare differential diagnosis of irreversible lung pathologies leading to failure of extracorporeal membrane oxygenation therapy for neonatal respiratory failure. PMID- 17914310 TI - Fusidic acid and heparin lock solution for the prevention of catheter-related bloodstream infections in critically ill neonates: a retrospective study and a prospective, randomized trial. AB - OBJECTIVE: Catheter-related bloodstream infections (CRBSIs) are one of the main morbidities in critically ill neonates. The objective of the present study was to assess the efficacy of a fusidic acid-heparin lock in the prevention of CRBSIs. DESIGN: A preliminary retrospective study showed that staphylococcal infections were largely prevalent. We planned a prospective, randomized trial to ascertain whether fusidic acid and heparin lock of central venous catheters would reduce the incidence of CRBSIs. SETTING: Level III neonatal intensive care unit. PATIENTS: One hundred three neonates were enrolled and randomly assigned to a treatment group (n = 50) or control group (n = 53). INTERVENTIONS: Fusidic acid (4 mg/mL) and heparin (10 IU/mL) lock in the treatment group. MEASUREMENTS AND MAIN RESULTS: The treatment group showed significantly lower incidence of CRBSIs (6.6 vs. 24.9 per 1000 catheter days; p < .01; relative risk 0.28; 95% confidence interval 0.13-0.60). No staphylococcal infections occurred in the treatment group, while in the control group Staphylococcus remained the main agent of CRBSI. Cost analysis comparing the present study and for the treatment of CRBSIs proved that antibiotic lock is financially favorable. CONCLUSIONS: Fusidic acid heparin lock solution reduced the incidence of CRBSIs in our neonatal intensive care unit. However, we recommend basing antibiotic lock on local CRBSI epidemiology. With regard to fusidic acid, further and broader studies could be useful to confirm our results. PMID- 17914311 TI - A history of adjunctive glucocorticoid treatment for pediatric sepsis: moving beyond steroid pulp fiction toward evidence-based medicine. AB - OBJECTIVES: To review the history of clinical use of corticosteroids with particular reference to adjunctive therapy for severe pediatric sepsis and, in this context, to provide an overview of what is known, what is not known, and what research questions are particularly relevant at this time. DATA SOURCE: Literature review using PubMed, cross-referenced article citations, and the Internet. CONCLUSIONS: The history of corticosteroid use in clinical medicine has been colorful, noisy, and always controversial. Therapeutic corticosteroid indications that initially seemed rational have frequently been refuted on closer, rigorous clinical trial inspection. Although it may be prudent to provide stress-dose steroids to children with septic shock who are clinically at risk for adrenal insufficiency (chronic or recent steroid use, purpura fulminans, etomidate or ketoconazole administration, hypothalamic, pituitary, adrenal disease), the safety and efficacy of stress-dose steroids as general adjunctive therapy for pediatric septic shock have not been established. Glucocorticoid administration does add potential risk to critically ill children. In particular, although adjunctive corticosteroids may hasten resolution of unstable hemodynamics in septic shock, this may occur at the metabolic cost of hyperglycemia. Clinical practice that fosters innovative therapy (off-label use) over research probably represents bad medical and social policy. Accordingly, pediatric critical care researchers have a responsibility to generate pediatric specific evidence-based medicine for adjunctive corticosteroid therapy for severe sepsis in children. PMID- 17914312 TI - The brave new psychiatry: beyond modernism, antimodernism and postmodernism in psychiatry. PMID- 17914313 TI - Three year outcomes of an early intervention for psychosis service as compared with treatment as usual for first psychotic episodes in a standard community mental health team - final results. AB - Sixty-two patients who had been treated for three years in an ad-hoc, assertive treatment team for patients who had suffered a first psychotic episode were compared to sixty-two patients who had been followed up after a first psychotic episode in a community mental health team. All patients had suffered a first or early psychotic episode. The main differences between the two teams was that the ad-hoc team was assertive in its approach, offered more structured psycho education, relapse prevention and psycho-social interventions, and had a policy of using atypical anti-psychotics at the lowest effective dose. There were many differences in outcome measures at the end of three years between the two groups. The EI patients are more likely to be taking medication at the end of three years. They are more compliant with medication. They are more likely to be prescribed Atypical Medication. The EI patients are more likely to have returned to Work or Education. The EI patients are more likely to remain living with their families. They are less likely to suffer depression to the extent of requiring anti-depressants. They appear to commit less suicide attempts. The patients in the EI service also appear to be less likely to suffer relapse and re hospitalisation, and are less likely to have involuntary admission to hospital. They have systematic relapse prevention plans based on Early Warning Signs. They and their families receive more psycho-education. These indications suggest that the EI patients are at the end of three years better able to manage their illness/vulnerability on their own than the CMHT patients. More patients in the EI group stopped using illicit drugs than in the CMHT group. All the above changes were statistically significant except for the total improvement in employment status and education status, which however approached significance. These results suggest that an ad-hoc Early Intervention Team is more effective than standard Community Mental Health Team in treating psychotic illness. PMID- 17914314 TI - Genetic factors in suicide: reassessment of adoption studies and individuals' beliefs about adoption study findings. AB - BACKGROUND: Based on multifaceted evidence (family, twin, adoption, molecular genetic, geographic, immigrant, and surname studies of suicide), the genetics of suicide is increasingly recognized. However, a comprehensive, stand-alone review of adoption studies in this area is unavailable, nor are there data on what people actually believe about adoption study findings. OBJECTIVES: To reassess the available adoption studies of suicide and to provide novel data on beliefs relating to these. METHOD: Key findings of the 3 existing adoption studies of suicide were reviewed and subjected to a meta-analysis. Beliefs pertaining to these were analyzed in 10 samples (total N = 1379) of different types (medical and psychology students, general population) from 6 countries. RESULTS: The combined evidence from adoption studies clearly supported genetic factors in suicide. Critical tests showed that suicide runs in the biological families of suicided (as well as schizophrenic or affectively ill) adoptees, but not in those of demographically matched healthy control adoptees alive (relative risk RR = 8.38, p = 0.0002). Individuals' beliefs about the plausibility of these findings were inconsistent and self-contradictory. Particularly, it was widely disbelieved that adoptees' risk of suicide resembles the one of their biological, not their adoptive, relatives (which is the case). CONCLUSIONS: Despite demonstrating a significant genetic effect, the evidence base from adoption studies of suicide is narrow. To broaden the evidence remains an important research agenda. Gaps in and deficiencies of this specific literature as well as general limitations of adoption study designs are discussed. PMID- 17914315 TI - The patient with schizophrenia and his father. The father's role and personality traits in the dynamics of the family with a son suffering from schizophrenia. AB - The role of the father in the onset and development of schizophrenic disorder/disturbance is very often secondary as compared to that of the mother. This study deals with the father's sphere in the family with a patient/son suffering from schizophrenia, the traits of his personality and his relationship with his son/patient, and with his wife/the mother of the patient. The methods applied were clinical interview, structured interview and semantic differential. The following statistical procedures were used: correlation, componential, discriminative, factor and quasicanonical analyses. The results include some characteristic excerpts from clinical interviews with patients/sons and their fathers, projective perception of current family relationships, quasicanonical analysis of the father's, mother's and son's experiences of the patient's/son's early childhood, and the projective view of the father. Among the conclusions reached, the phenomenon of "dead father" is emphasized as one of the main factors in the onset and development of schizophrenic disorder/disturbance. PMID- 17914316 TI - The influence of religious moral beliefs on adolescents' mental stability. AB - AIM: The aim of this paper is to determine the influence of religious moral beliefs on the stability of adolescents' mental health. METHODS: The sample consists of 240 mentally and physically healthy male and female adolescents attending a high school, who are divided into groups equalized by gender (male and female), age (younger 15, older 18 years); school achievement (very good, average student); behaviour (excellent, average); family structure (complete family with satisfactory family relations), and level of exposure to psycho social stress (they were not exposed to specific traumatizing events). Subjects were assessed with regard to the level of belief in some basic ethical principles that arise from religious moral values. The score of religious moral belief index was used to compare two groups of subjects. For sample selection the measuring instruments were used to assess the religious, moral and social profile of subject. For the assessment of personality structure a standardized test battery (Freiburg's Personality Questionnaire/ Das Freiburger Personlichkeitsinventar - FPI, Profile Index of Emotions - PIE, Life Style Questionnaire - OM) was used to assess personality profile, emotional profile and subject's defence orientation. RESULTS: The score of the moral belief index was negatively correlated to neuroticism and depressiveness (Pearson's r=-0.242, P<0.001; r=-0.311, P<0.001, respectively). Spontaneous and reactive aggressiveness and irritability were negatively correlated with the score of moral belief index (Pearson's r=-0.197, P=0.002; r=-0.147, P=0.023; r=-0.350, P<0.001, respectively). Emotional instability is negatively associated with the moral belief index of the investigated adolescents (Pearson's r=-0.324, P<0.001). The moral belief index was highly negatively correlated with repression (r=-0.206, P=0.001), regression (r=-0.325, P<0.001), compensation (r=-0.186, P=0.004), transfer (r=-0.290, P<0.001) and defensive orientation (r=-0.129, P=0.046). Verified intellectualisation and reactive formation are in positive correlation with the moral belief index among our investigated adolescents (Pearson's r=0.168, P=0.009; r=0.356; P<0.001, respectively). CONCLUSIONS: A higher index of religious moral beliefs in adolescents enables better control of impulses, providing better mental health stability. It enables neurotic conflicts typical for adolescence to be more easily overcome. It also causes healthier reactions to external stimuli. A higher index of religious moral beliefs of young people provides a healthier and more efficient mechanism of anger control and aggression control. It enables transformation of that psychical energy into neutral energy which supports the growth and development of personality, which is expressed through socially acceptable behaviour. In this way, it helps growth, development and socialization of the personality, leading to the improvement in mental health. PMID- 17914317 TI - Involuntary emotional expression disorder - new/old disease in psychiatry and neurology. AB - Involuntary emotional expression disorder (IEED) is underrecognized by clinicians, misdiagnosed as depression or bipolar disorder and undertreated, because clinicians are unfamiliar with the disorder. An important clinical consideration for IEED is that of distinguishing mood from affect. IEED describes a syndrome of relatively stereotypical episodes of uncontrollable crying and/or laughing, resulting from lesions of multiple types, in multiple brain regions, without an apparent stimulus to trigger such responses. This syndrome is common among a number of neurological diseases like patients with a stroke or traumatic brain injury (TBI), patients with amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), as well as dementias such as Alzheimer;s disease (AD), and motor disorders such as Parkinson;s disease (PD). The neuropathological cause and neurochemistry of the disorder remains unclear. There is general agreement that IEED is the result of an injury to the neurological pathways that control the expression of emotions. Adequate treatment can reduce the frequency and improve the quality of life of patients and caregivers. PMID- 17914318 TI - Efficacy of risperidone treatment in Smith-Magenis syndrome (del 17 pll. 2). AB - Smith-Magenis syndrome (SMS) is a clinically recognizable multiple congenital anomaly and mental retardation syndrome caused by an interstitial deletion of chromosome 17 pll.2. Although the physical and molecular genetic features of SMS are increasingly well understood, work is more limited on SMS's behavioral phenotype, which includes self-injury, tantrums, aggression, attention deficit, and sleep disturbance. This case-report describes the lowering of the aggression level of a 13 year old individual with SMS. PMID- 17914319 TI - Contemporary psychopharmacotherapy in the context of brave new psychiatry, well being therapy and life coaching. AB - Despite modern advances in clinical psychopharmacology, mental disorders are still the leading cause of disability all over the world. Drug treatment of mental disorders is unfortunately too often associated with partial remissions, frequent relapse or recurrence as well as with persistent residual symptoms, distress and low levels of well-being, life-satisfaction and quality of life. The practical failure of psychopharmacotherapy to improve life-satisfaction and well being is related to treatment focus on psychopathology and decrease of illness and not on the development of wellness, purpose of life and creativity. NLP, life coaching and well-being therapy may significantly increase effectiveness of psychopharmacotherapy shaping an efficient and meaningful treatment context for achieving a full recovery. PMID- 17914320 TI - Psychoactive drugs and neuroplasticity. AB - The influence of psychoactive drugs on neuroplasticity, especially on neurogenesis is reviewed. From psychopharmacological point of view most interesting results are those showing neurogenesis that neurogenesis is increased by SSRI. However, the role of serotonin system in neurogenesis as well as significance of neurogenesis in the beneficial effect of psychotropic drugs requires a lot of additional and new inventive research. PMID- 17914321 TI - Pharmacotherapy for the soul and psychotherapy for the body. AB - The "mind-body" dualism has shaped the development of psychiatry. At the very beginning psychiatry was related to the mind and the rest of the medicine to the body. The main reasons for such division were lack of biological evidence for psychiatric disorders and wrong beliefs about demonic origins of "lunacy". But although the development of science offered more than enough biological evidence to understand brain as an organ of origin for psychiatric disorders, the dualism of mind and body remained alive even in the modern classification systems. One of the consequences was another dualism that differ biological (e.g. pharmacotherapy) from psychological therapy (e.g. psychotherapy) as completely different approaches. The purpose of this article is to offer enough evidence to reframe the existing dualisms into a different paradigm. In every illness both mind and body can be affected to a different extent. Which part of an illness is body and which part is mind is often difficult to differentiate even when we compare a person with broken leg with a person with acute stress reaction. For that reason it might be an over-simplification to differentiate sharply between biological and psychological therapies. The evidence show that psychotherapy influences biology of the brain and that pharmacotherapy influences psychological, social and developmental dimensions of the diseased person as well as overall well-being and functionality. In the era where medicine discovered psychology and psychiatry discovered biology, the debates and divisions that steam out of past dualisms should end. Every practising physician regardless of the medical discipline uses in everyday practice both biological and psychological approaches to help the patient successfully. PMID- 17914322 TI - Ethics of the early intervention in the treatment of schizophrenia. AB - When second generation antipsychotics were introduced in the mid 1990-s they offered the possibility of early psychopharmacological interventions in the treatment of schizophrenia. The idea applying antipsychotics in the prodromal phase of schizophrenia today is an realistic option. However ethical dilemmas about offering antipsychotics to the adolescents with at risk mental states, of whom only a few are real prodromes of schizophrenia remain for clinicians. In the literature about the ethics of the early interventions in psychiatry there are still many ethical questions which call for caution because of the low predictive value of at risk mental states, of which only 40% turn out to be a real prodrome of schizophrenia. These ethical questions can be addressed in three categories: - how to best identify who should receive early pharmacological intervention? - what this intervention should consist of? - how to evaluate treatment efficacy in the absence of illness base rates in the adolescents with a real prodrome of schizophrenia? Besides, arguing against the concept of early psychopharmacological interventions in the adolescent population are the fact of the unknown effect of antipsychotics on the developing brain as well as negative effects of stigma on those adolescents who receive them. The authors in the article analyse these ethical questions and take the side of those clinicians who think that caution and careful ethical judgment are needed before the prescribing of antipsychotics to adolescents with at risk mental states. PMID- 17914323 TI - Psychopharmacotherapy and spirituality. AB - Although the connection between spiritual and physical has been acknowledged since the oldest of human civilizations and emphasized in almost all of religions, It has taken a lot of time for that connection to gradually recover its lost meaning. As it is evident that many diseases and illnesses can not be explained purely by physical causes nor treated with purely physical methods there is a growing interest in spirituality and its usability in the treatment of various diseases and states, as well as in everyday life. Despite the fact that a sense of positive, nourishing and healing power of faith is deeply rooted in every religion, objective and empirical research of that connection has been avoided for centuries, and those studies which were conducted are only rudimentary, on the outskirts of empirical science. Scientific literature regarding spirituality and mental health points to a conclusion that spirituality and faith are positively correlated with positive therapeutic outcome and the possible explanation for that can be found in the fact that spirituality and religiosity can satisfy some of the basic needs of psychiatric patients. Efficacy of psychiatric treatment improved with the introduction of psychotropic medicaments and psychotherapeutic techniques, but the outcome is still not satisfying because relapse, recidivation and discontinuation of therapy occur very often. On the other hand, spirituality and religiosity play a very significant role in the healing process because they provide people with strength and will to fight their problems and disease itself. Psychopharmaceuticals are evidently irreplaceable in the therapy of mental disorders, but they are only one of the segments of the overall therapy. In order to adequately answer to the spiritual and religious needs of their patients, psychiatrists, psychologists and related health care professionals are faced with the need for expanding scientific concepts which served as the basis for development of many psychiatric methods and techniques. PMID- 17914324 TI - Pharmacogenomics of mental illnesses: do sex-specific differences matter? AB - OBJECTIVES: Genetic factors are extensively studied in respect to drug response in psychiatric disorders. Recent evidence suggests that action of reproductive steroid hormones in brain may also have a role. METHODS: Sex-specific differences in terms of illness onset, duration, severity of symptoms and treatment response are well documented. It is believed they result from different brain morphology and function between sexes, factors being highly influenced by sex hormones. RESULTS: The synergistic effects of the genetic background and potential prenatal stressors may influence the processes of sexual brain maturation, the most vulnerable period for developing susceptibility for psychiatric disorders. A resulting neuroendocrine dysfunction implies inadequate response of brain structures to pubertal flow of circulating sex hormones. CONCLUSION: Steroid sex hormones, at least estrogen, are major parts of the communication system in the brain. For that reason, estrogen receptors could be attractive targets in development of new treatment strategies. Potential benefits from compounds mimicking estrogen should be also considered in clinical practice. PMID- 17914325 TI - Pharmacogenetics in modern psychiatry. AB - There is no unique scientific agreement about genetic variations that could beyond any doubt predict phenotype of the response to pharmacotherapy. The knowledge about the predictors for therapy and serious adverse effects could be very useful in clinical practice. It is obvious that the combination of variation in metabolizing enzymes, transporter proteins and drug targets fully explains heterogeneity in response to psychiatric treatment. We assume that genetic polymorphisms of serotoninergic transporter and MDR1 could be important in prediction of therapeutic response in patients with major depression treated with paroxetine. Our results show that SERTPR-LL genotype might predispose significantly better paroxetine treatment response compared to SS genotype in MDD patients and that variants G2677T and C3435T are not associated with therapeutic response to paroxetine in patients with major depressive disorder. PMID- 17914326 TI - Use of non-invasive neuroradiological methods in research of psychoactive drugs. AB - Non-invasive neuroradiological methods like magnetic resonance spectroscopy (MRS), functional magnetic resonance imaging (fMRI), blood oxgenation level dependent (BOLD) imaging recently entered into the areas of research in psychiatry, psychoactive drug development, as well as in clinical practice. fMRI can identify the regions of the brain associated with various functions, can monitor recovery, progression, and response to treatment and MRS can be used to study brain chemistry and metabolism. BOLD-imaging provides an indirect indication of neuronal activity. Future developments of different neuroimaging techniques are promising not only in surgical planning, functional assessment in brain tumor management, monitoring functional changes, but also in discovery of pathophysiology of psychiatric disorders and recognition of new pharmacological targets. Those techniques could be implemented in the process of drug discovery and identification of biomarkers which are clinically relevant for development of candidate drugs. Furthermore, those techniques establish the bridge between preclinical and clinical studies and allow the drug research in human in vivo. PMID- 17914327 TI - Antidepressants and suicidality: the basis of controversies. AB - Regulatory agencies of different European countries and the United States have been critically examining the possible link between suicidality and antidepressant use in children and adults, which has resulted in an FDA directive to the manufacturers of all antidepressant medications to add a 'black box' warning. 'Black box' warning describes the increased risk of suicidality in persons who take antidepressants. Because the news media's coverage of the antidepressant-suicide controversy has increased dramatically in the last few years, serious concerns need to be performed. In this review, the possible risk benefit ratio has been estimated according to the use of selective serotonin reuptake inhibitors (SSRIs) antidepressants due to relevant psychobiological, clinical and epidemiologic data. PMID- 17914328 TI - Herbal medicine in the treatment of mental disorders. AB - During five thousand years of recorded history, we know that from the ancient times people have used different methods and procedures in treatment of different psychiatric disorders and very often these were medicinal preparations of plants. Numerous scientific discoveries and industrial age gave a big contribution to medicine development and significantly improved quality of life for psychiatric patients during the last century. However, evidence-based medicine after big bliss faced a lot of disappointments, and an attitude that some natural drugs were unnecessarily thrown out of use step by step came along. On the other hand, there are a huge number of patients that use natural medicinal plants in self treatment of different psychiatric disorders. The aim of this article is not giving advantage to natural medicines over chemicals, nor to support self treatment. The aim is to encourage thinking about the meaning of natural medicines in the treatment of mental disorders and an attempt of preventing to push them out totally into silence wherever it is justified and based on facts. PMID- 17914329 TI - Physical activity among adults with a disability--United States, 2005. AB - The health benefits of physical activity have been well documented and are supported by recommendations from Healthy People 2010 (focus area 22); however, fewer than half of U.S. adults follow these recommendations. Physical inactivity is particularly prevalent among adults with a disability, who are at increased risk for functional limitations and secondary health conditions (e.g., obesity, depression, or social isolation) that can result from their disabilities, behavior, lifestyle, or environment. To estimate the state-specific prevalence of physical activity and physical inactivity among adults with and without a disability, CDC analyzed data from the 2005 Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the results of that analysis, which determined that, compared with adults without a disability, a smaller proportion of adults with a disability met national recommendations for physical activity (37.7% versus 49.4%), and a greater proportion were physically inactive (25.6% versus 12.8%). Public health measures to promote and increase physical activity should include consideration for the needs of adults with disabilities. PMID- 17914330 TI - Salmonella Oranienburg infections associated with fruit salad served in health care facilities--northeastern United States and Canada, 2006. AB - During June-July 2006, a total of 41 culture-confirmed Salmonella serotype Oranienburg infections were diagnosed in persons in 10 northeastern U.S. states and one Canadian province. This report describes the epidemiologic, environmental, and laboratory investigations of this outbreak by federal, state, and local health agencies; the Food and Drug Administration (FDA); and the Canadian Food Inspection Agency. The results of the investigations determined that illness was associated with eating fruit salad in health-care facilities. Although the fruit salads were produced by one processing plant, the source of contamination was not determined. This outbreak highlights the importance of laboratory-based surveillance of Salmonella, including molecular subtyping, and timely communication of public health information. PMID- 17914331 TI - Progress in measles control--Nepal, 2000-2006. AB - In 2002, the United Nations General Assembly Special Session on Children set a goal to reduce global measles deaths by half (compared with 1999) by 2005. Nepal, a southeast Asian country with an estimated population of 27 million, adopted the measles mortality reduction strategies of the World Health Organization (WHO) in 2003, with a goal of reducing measles deaths by half (compared with 2003) by 2005. The strategies consisted of strengthening routine childhood immunization programs, providing a second opportunity for measles vaccination through supplementary immunization activities (SIAs), improving surveillance, and improving measles case management. This report describes routine immunization activities in Nepal, the implementation of measles SIAs, and measles surveillance data for the period 2000-2006. The findings demonstrate a substantial decrease in reported measles incidence. Assuming a reduction in measles deaths that paralleled the decrease in incidence, the findings also suggest progress toward the goal of measles mortality reduction. PMID- 17914332 TI - High-risk benign breast lesions: current strategies in management. AB - BACKGROUND: High-risk benign breast lesions can create confusion for both the patient and the clinician. This paper reviews the characteristics of these lesions to help direct appropriate management. METHODS: The authors reviewed the literature regarding high-risk breast lesions and include management guidelines that we employ at our institute. RESULTS: High-risk breast lesions offer varying degrees of increased risk for the future development of breast cancer. Chemoprevention may be used to help decrease the risks from some lesions. CONCLUSIONS: The management of high-risk benign breast lesions can be confusing. Clinicians should assess the risk of future breast cancer and develop a proper screening and prevention strategy for each individual patient. PMID- 17914333 TI - Management options after prophylactic surgeries in women with BRCA mutations: a review. AB - BACKGROUND: Although breast cancer is relatively common, only about 5% of cases are due to inheritance of highly penetrant cancer susceptibility genes. The majority of these are caused by mutations in the BRCA1 and BRCA2 genes, which are also associated with an increased risk of ovarian cancer. Increased surveillance, chemoprevention, and prophylactic surgeries are standard options for the effective medical management of mutation carriers. However, optimal management of female carriers who choose to undergo prophylactic surgeries is still poorly understood. METHODS: The authors provide an overview of the current literature regarding medical management options for women carriers of BRCA1 and BRCA2 gene mutations and the implications for those individuals who have chosen to undergo prophylactic surgeries. RESULTS: BRCA mutation carriers who opt for prophylactic surgeries are still at risk for development of malignancy, and appropriate monitoring is warranted. CONCLUSIONS: There are limited data on the appropriate medical management for BRCA mutation carriers after prophylactic surgeries. However, a management plan can be extrapolated from the general management recommendations for surveillance and other risk-reducing strategies in BRCA positive individuals. PMID- 17914334 TI - Recent advances in breast-specific imaging. AB - BACKGROUND: Imaging the breast is a vital component not only for breast cancer screening, but also for diagnosis, evaluation, treatment, and follow-up of patients with breast cancer. METHODS: The author reviews recent advances and also provides her personal experience in describing the status of digital mammography, computer-aided detection, dedicated magnetic resonance imaging (MRI), and positron-emission mammography for evaluating the breast. RESULTS: Full-field digital mammography is superior to standard mammography in women under 50 years of age and in those with dense breasts. Computer-aided detection assists inexperienced mammographers and enhances detection of microcalcifications in dense breasts. Breast MRI is useful in preoperative evaluation, clarification of indeterminate mammograms, and follow-up of BRCA mutation carriers. The specificity of MRI remains problematic, however. Positron-emission mammography promises enhanced detection of ductal carcinoma in situ (DCIS), even when not associated with microcalcifications, and should aid surgical planning. CONCLUSIONS: These four significant advances in breast imaging have all improved the sensitivity of detecting breast abnormalities. Cost issues, however, may limit the widespread application of these advances. PMID- 17914335 TI - Breast ductal secretions: clinical features, potential uses, and possible applications. AB - BACKGROUND: Nipple discharge accounts for approximately 5% of visits to a breast specialist surgical practice and may be encountered as the chief complaint by many other types of physicians. The vast majority of breast cancers originate in the ductal system, which prompted interest in the evaluation of the intraductal approach to breast cancer. Ductoscopy, nipple aspiration, and ductal lavage have emerged as innovative fields of study that may have clinical applications. METHODS: We performed a literature search of published manuscripts using the keywords nipple discharge, breast ductal secretions, and intraductal approach. We also report our single-institution experience in managing nipple discharge. RESULTS: We present our institutional algorithm for the management of nipple discharge. The possible etiologies of nipple discharge and the appropriate workup are reviewed. Three evolving minimally invasive techniques for the evaluation of high-risk patients include ductoscopy, nipple aspiration, and ductal lavage. Nipple aspiration and ductal lavage fluid may be assayed for cytology, genomic, gene expression, and proteomic studies. Several different translational approaches are being undertaken to investigate the local microenvironment associated with the development and progression of breast carcinoma. CONCLUSIONS: Nipple aspiration fluid and ductal lavage offer the opportunity to study the local microenvironment of the ductal system, which is where most breast cancers originate. These powerful approaches to biomarker analysis could be applied to the prevention and treatment of breast cancer. PMID- 17914336 TI - Proteomic approach to breast cancer. AB - BACKGROUND: Breast cancer is the most common cancer affecting women worldwide. Despite tremendous advances in screening, diagnosis, and treatment, the causes of this disease remain elusive and complex. Proteomics is a rapidly developing field that can explore the heterogeneity of breast cancer and supplement the wealth of information gained from genomics. METHODS: This article serves as an overview of the application of matrix-assisted laser desorption/ionization source with a time of-flight (MALDI-TOF) proteomic techniques as applied to breast cancer. Examples of the clinical applicability of MALDI-TOF mass spectrometry are provided but represent only a fraction of the potential uses yet to be discovered. In addition, a brief summary of the bioinformatics issues that surround proteomics is included. RESULTS: Mass spectrometry has provided new proteomic approaches to unravel the complexities of clinical specimens relevant to breast cancer diagnostics. In particular, MALDI-TOF mass spectrometry analysis has been used to differentiate cancer profiles from benign profiles in samples from sera, plasma, tissue, nipple fluid, and ductal lavage. Some discriminating proteins have subsequently been identified. CONCLUSIONS: Mass spectrometry applications to breast cancer diagnostics continue to be developed but are evolving faster than bioinformatics/statistical analysis can adapt. The future of these techniques in terms of clinical investigation is limitless, but in terms of general applicability, these applications are currently cost-prohibitive. PMID- 17914337 TI - Health disparities in receipt of screening mammography in Latinas: a critical review of recent literature. AB - BACKGROUND: Increased use of screening mammography is associated with lower death rates from breast cancer in the United States. Despite recommendations that women over 40 years of age should obtain regular screening mammography at least every 2 years, many women do not adhere to these guidelines. Historically, women from underserved and minority populations have been less likely to receive screening mammography. METHODS: A critical review of recent research literature was conducted to evaluate whether Latinas are less likely to receive screening mammography, determine whether disparities in screening mammography persist when controlling for other variables, and examine what other variables are associated with screening mammography. The articles were obtained from a search of the PubMed database. RESULTS: Fifteen published articles met the inclusion criteria and were critically reviewed. The unadjusted odds ratios (ORs) of the association between Hispanic ethnicity and screening mammography ranged from 0.40 to 0.93. For the most part, the ORs adjusted for other variables in multiple logistic regression analyses increased (range: 0.3 to 1.67). Age, education, income, health insurance, having a usual source of care, and having a recent visit to a physician were consistently related to screening mammography in multiple logistic regression analysis. CONCLUSIONS: Hispanic ethnicity is a risk factor for lack of adherence to screening mammography. However, other demographic, socioeconomic, and health system variables account for some of the disparity related to Hispanic ethnicity. PMID- 17914338 TI - Educational intervention for women undergoing image-guided breast biopsy: results of a randomized clinical trial. AB - BACKGROUND: The process of informed consent has been examined for patients undergoing various procedures but not breast biopsy. Our study was a randomized trial that examined the effect of an educational flip chart as part of the informed consent. METHODS: A total of 122 patients referred for stereotactic or ultrasound-guided core breast biopsy were randomly assigned to receive an informed consent discussion with or without an illustrated flip chart. The chart included information about breast anatomy, pathology, and diagnostic procedures. Outcome measures included objective knowledge, subjective knowledge, anxiety, and satisfaction. RESULTS: Analysis showed few significant main effects of the intervention. However, results showed interactions between experimental condition and race/ethnicity, indicating that the intervention was effective in enhancing objective and subjective knowledge for African American but not Caucasian patients. Anxiety after consultation was higher among patients assigned to the flip chart condition, possibly because they were better informed about the risks of the procedure. Patients who underwent biopsy sooner after learning they needed one were more satisfied with their care. CONCLUSIONS: The usual care consent process is effective for many but not all patients. Informed consent that employs visual aids may help overcome characteristics of the consent process that are ineffective for some patients. PMID- 17914339 TI - The impact of education and literacy levels on cancer screening among older Latin American and Caribbean adults. AB - BACKGROUND: There is limited information related to the effects of education and literacy on cancer screening practices among older adults in Latin American and Caribbean countries. METHODS: To determine the association between education and cancer screening use, we developed a cross-sectional study using data from the Health, Well-Being and Aging in Latin America and the Caribbean Study. The sample included 4,183 men and 6,708 women aged 60 years and older from seven cities. The outcomes are mammogram and Pap smear use in women and prostate examination use in men within the last 2 years. RESULTS: In general, illiterate or lower-educated older men and women have the lowest rates of cancer screening use compared with higher-educated counterparts. Multivariate logistic models, by city and in a combined sample of six cities showed that high education is associated with higher odds of having a mammogram or a Pap smear in women and a prostate examination in men. CONCLUSIONS: Older adults with low educational or literacy levels should be targeted for screening programs in these populations. PMID- 17914340 TI - Factors affecting uptake of cervical cancer screening among clinic attendees in Trelawny, Jamaica. AB - BACKGROUND: Use of the Pap test has resulted in a decline in cervical cancer mortality in developed countries. Yet, despite established cervical cancer screening programs, a significant portion of Jamaican women are not undergoing screening for cervical cancer. This study was carried out to identify factors that affect Jamaican women's decisions to screen for cervical cancer. METHODS: A population survey was administered to 367 clinic-attending women 25 to 54 years of age in the Parish of Trelawny from May to July of 2005. An interviewer administered questionnaire assessed the women's knowledge, attitudes, and practices regarding cervical cancer and cervical cancer screening. RESULTS: Overall, 11% of the women had never had a Pap smear and only 38% had a Pap test within the last year. Annual visits to a health provider have a strong influence on women's decisions to regularly screen for cervical cancer. Provider recommendation also positively affected initial receipt of a Pap smear as well as continued regular screening. CONCLUSIONS: Programs that promote annual health checkups, encourage consistent provider recommendations, and emphasize screening as a preventive measure might positively influence women's decisions to screen for cervical cancer. PMID- 17914341 TI - Spinal radiosurgery for metastatic disease of the spine. AB - BACKGROUND: Metastatic tumor in the spinal column is common, causing symptomatic spinal cord compression in approximately 25,000 patients annually. Although surgical treatment of spinal metastases has become safer, less invasive, and more efficacious in recent years, there remains a subset of patients for whom other treatment modalities are needed. Stereotactic radiosurgery, which has long been used in the treatment of intracranial lesions, has recently been applied to the spine and enables the effective treatment of metastatic lesions. METHODS: We review the evolution of stereotactic radiosurgery and its applications in the spine, including a description of two commercially available systems. RESULTS: Although a relatively new technique, the use of stereotactic radiosurgery in the spine has advanced rapidly in the past decade. Spinal stereotactic radiosurgery is an effective and safe modality for the treatment of spinal metastatic disease. CONCLUSIONS: Future challenges involve the refinement of noninvasive fiducial tracking systems and the discernment of optimal doses needed to treat various lesions. Additionally, dose-tolerance limits of normal structures need to be further developed. Increased experience will likely make stereotactic radiosurgery of the spine an important treatment modality for a variety of metastatic lesions. PMID- 17914342 TI - You can't cure a disease by throwing money at it, can you? PMID- 17914343 TI - Skin biopsy as a diagnostic tool in peripheral neuropathy. AB - Skin biopsy is a safe, minimally invasive, painless and cheap tool for providing diagnostic information on small nerve fibers, which are invisible to routine neurophysiological tests. Biopsy can be performed in hairy skin to investigate unmyelinated and thinly myelinated fibers and in glabrous skin to examine large myelinated fibers. Morphometric analysis of skin nerves is readily accomplished through the use of immunohistochemical techniques, and has proved to be reliable, reproducible and unaffected by the severity of neuropathy. One further advantage of skin biopsy over conventional nerve biopsy is that it allows somatic nerve fibers to be distinguished from autonomic nerve fibers. Morphological changes, axonal degeneration and abnormal regeneration occur in cutaneous nerves very early in the course of peripheral neuropathies, making skin biopsy a promising tool for investigating the progression of neuropathy and the effect of neuroprotective treatments in clinical practice and trials. This article reviews the techniques that are used to investigate the innervation of human skin, the possible uses of skin biopsy in diagnosing and monitoring peripheral neuropathies, and correlations between skin biopsy findings and those of other diagnostic methods. PMID- 17914345 TI - Mechanisms of disease: basic-research-driven investigations in humans--the case of hyperkinetic disorders. AB - Recent studies have revealed that animal models of Huntington's disease and levodopa-induced dyskinesia show similar changes in corticostriatal transmission. In both conditions, corticostriatal long-term potentiation (LTP) is abnormally stable, causing persistent inhibition of the output nuclei of the basal ganglia, and involuntary movements. This finding has led to speculation that interventions that interfere with the maintenance of corticostriatal LTP might be useful as antidyskinetic treatments. One approach that is known to reduce cortical hyperexcitability in human disorders is the delivery of a long train of low frequency ( approximately 1 Hz) stimulation using repetitive transcranial magnetic stimulation. This stimulation protocol is remarkably similar to that used in brain slices to reverse previously induced LTP, and consequently it has been used in patients with Huntington's disease or levodopa-induced dyskinesia to try to interfere with the abnormal corticostriatal plasticity postulated to underlie the motor disturbances in these conditions. Clear antidyskinetic effects of stimulation have been obtained in both disorders, without significant side effects. These findings could have implications for the treatment of hyperkinetic disorders in the clinic, and they illustrate how basic neuroscience can generate predictions to be tested in patients. PMID- 17914346 TI - A case of celiac disease mimicking amyotrophic lateral sclerosis. AB - BACKGROUND: A 44-year-old male presented to a general neurology clinic with a 6 month history of progressive right-sided spastic hemiparesis without sensory symptoms or signs. The thigh muscle in the affected leg showed signs of wasting. The patient had a remote family history of celiac disease. INVESTIGATIONS: Neurological examination, neurophysiological studies, brain MRI scan, routine blood tests, duodenal biopsy, cerebrospinal fluid analysis including polymerase chain reaction test for JC virus DNA, serological testing for HIV and for the presence of serum antibodies to endomysium, gliadin and tissue transglutaminase. DIAGNOSIS: Celiac disease with neurological involvement, mimicking amyotrophic lateral sclerosis. MANAGEMENT: Strict gluten-free diet. PMID- 17914344 TI - Injury and recovery in the developing brain: evidence from functional MRI studies of prematurely born children. AB - Functional MRI (fMRI) might provide important insights into emerging data that suggest that recovery from injury can occur in the brains of children born prematurely. Strategies employing auditory stimulation demonstrate blood-oxygen level-dependent (BOLD) activation in preterm infants as young as 33 weeks' gestational age, and reliable BOLD signal in response to visual stimulation occurs at term-equivalent age. Strategies based on fMRI are particularly suited to the study of language and memory, and emerging data are likely to provide insights into perplexing reports that have demonstrated improving cognitive scores but persistent volumetric and microstructural changes in frontotemporal language systems in the prematurely born. Even when sex, gestational age and early medical and environmental interventions are taken into account, fMRI data from several investigators suggest the engagement of alternative neural networks for language and memory in the developing preterm brain. PMID- 17914347 TI - Neurology residency training in the US and Poland. PMID- 17914348 TI - CCN3 is a novel endogenous PDGF-regulated inhibitor of glomerular cell proliferation. AB - CCN proteins affect cell proliferation, migration, attachment, and differentiation. We identified CCN3 as a suppressed gene following platelet derived growth factor (PDGF)-BB or -DD stimulation in a cDNA-array analysis of mesangial cells. In vitro growth-arrested mesangial cells overexpressed and secreted CCN3, whereas the addition of the recombinant protein inhibited cell growth. Induction of mesangial cell proliferation by PDGF-BB or the specific PDGF beta-receptor ligand PDGF-DD led to downregulation of CCN3 mRNA, confirming the array study. Specific PDGF alpha-receptor ligands had no effect. CCN3 protein was found in arterial smooth muscle cells, the medullary interstitium, and occasional podocytes in the healthy rat kidney. Glomerular CCN3 was low prior to mesangial proliferation but increased as glomerular cell proliferation subsided during mesangioproliferative glomerulonephritis (GN). Inhibition of PDGF-B in mesangioproliferative disease led to overexpression of glomerular CCN3 mRNA. CCN3 localized mostly to podocytes in human glomeruli, but this expression varied widely in different human glomerulonephritides. Glomerular cell proliferation negatively correlated with CCN3 expression in necrotizing GN. Our study identifies CCN3 as an endogenous inhibitor of mesangial cell growth and a modulator of PDGF-induced mitogenesis. PMID- 17914350 TI - Apical junctional complexes and cell polarity. AB - Recent studies have greatly expanded our knowledge of initial events that lead to epithelial cell polarity. Epithelial polarity is defined, in part, by apical cell cell tight junctions that separate the plasma membrane into the apical domain and the basolateral domain, as well as the zonula adherens that mediate intercellular adhesion. The process of epithelial polarization is closely coupled to the biogenesis of these junctions. Studies in mammalian epithelial cells and lower organisms have identified two evolutionarily conserved junctional complexes as important epithelia polarity regulators: the Crumbs complex and the partitioning defective complex. Disruption of the components of the two complexes leads to a disorder of epithelial cell polarity and defects in junction formation or maintenance. Recent discoveries have revealed more details of how the two junctional polarity complexes function to establish epithelial polarity. They also raised the question about the relationship between polarity and adhesion. Although it is widely accepted that cell-cell adhesion provides a landmark from which polarity can proceed, there are results pointing to the possibility that polarity complexes can regulate cell-cell adhesion. It seems likely that proteins that control cell adhesion and cell polarity work intimately together to establish final epithelial polarity. PMID- 17914349 TI - Renal response to metabolic acidosis: role of mRNA stabilization. AB - The renal response to metabolic acidosis is mediated, in part, by increased expression of the genes encoding key enzymes of glutamine catabolism and various ion transporters that contribute to the increased synthesis and excretion of ammonium ions and the net production and release of bicarbonate ions. The resulting adaptations facilitate the excretion of acid and partially restore systemic acid-base balance. Much of this response may be mediated by selective stabilization of the mRNAs that encode the responsive proteins. For example, the glutaminase mRNA contains a direct repeat of 8-nt AU sequences that function as a pH-response element (pHRE). This element is both necessary and sufficient to impart a pH-responsive stabilization to chimeric mRNAs. The pHRE also binds multiple RNA-binding proteins, including zeta-crystallin (zeta-cryst), AU-factor 1 (AUF1), and HuR. The onset of acidosis initiates an endoplasmic reticulum (ER) stress response that leads to the formation of cytoplasmic stress granules. zeta cryst is transiently recruited to the stress granules, and concurrently, HuR is translocated from the nucleus to the cytoplasm. On the basis of the cumulative data, a mechanism for the stabilization of selective mRNAs is proposed. This hypothesis suggests multiple experiments that should define better how cells in the kidney sense very slight changes in intracellular pH and mediate this essential adaptive response. PMID- 17914351 TI - Randomized controlled study of biocompatible peritoneal dialysis solutions: effect on residual renal function. AB - Residual kidney function is important for patient and technique survival in peritoneal dialysis (PD). Biocompatible dialysis solutions are thought to improve function and viability of peritoneal mesothelial cells and to preserve residual renal function (RRF). We conducted a randomized controlled study comparing use of biocompatible (B) with standard (S) solutions in 93 incident PD patients during a 1-year period. The demographics, comorbidities, and RRF of both groups were similar. At 3 and 12 months, 24-h urine samples were collected to measure volume and the mean of urea and creatinine clearance normalized to body surface area. Surrogate markers of fluid status, diuretic usage, C-reactive protein concentration, peritonitis episodes, survival data, and peritoneal equilibrium tests were also collected. Changes in the normalized mean urea and creatinine clearance were the same for both groups, with no significant differences in secondary end points. Despite non-randomized studies suggesting benefits of these newer biocompatible solutions, we could not detect any clinically significant advantages. Additional studies are needed to determine if advantages are seen with longer term use. PMID- 17914353 TI - The role of mitochondria in oxidative and nitrosative stress during ischemia/reperfusion in the rat kidney. AB - Reoxygenation following ischemia causes tissue oxidative stress. We studied the role of oxidative stress caused by kidney ischemia/reperfusion (I/R) on the mitochondria of renal tissue slices. I/R caused the mitochondria to be swollen, fragmented, and have lower membrane potential. The mitochondria generated more reactive oxygen species (ROS) and nitric oxide (NO) in situ as measured by fluorescence of ROS- and NO-sensitive probes. Infusion of lithium ion, an inhibitor of glycogen kinase synthase-3, caused phosphorylation of its Ser-9 and restored the membrane potential and decreased ROS production of the mitochondrial fraction. Ischemic kidney and hypoxic rat preconditioning improved mitochondrial membrane potential and lowered ROS production caused by subsequent I/R similar to lithium ion infusion. Preconditioning normalized NO production in mitochondria as well. The drop in the mitochondrial membrane potential was prevented by NO synthase inhibition, demonstrating a strong contribution of NO to changes in mitochondrial energy metabolism during the I/R transition. Mitochondria in the I/R-stressed kidney contained less cytochrome c and more pro-apoptotic Bax, consistent with apoptotic degradation. PMID- 17914352 TI - Heparan sulfate domains on cultured activated glomerular endothelial cells mediate leukocyte trafficking. AB - Heparan sulfate (HS) proteoglycans by playing key roles in the leukocyte endothelial interactions are thought to mediate inflammatory cell influx in proliferative glomerulonephritis. Here, we evaluated the specific features within glomerular endothelial HS that promote leukocyte adhesion. Mouse and human glomerular endothelial cells activated by tumor necrosis factor (TNF)-alpha or interleukin (IL)-1beta increased expression of inflammatory N- and 6-O-sulfated HS domains. In addition, altered expression of HS-modifying enzymes occurred, a feature also found in mouse kidneys with anti-glomerular basement membrane disease or lupus nephritis. Inhibition of the nuclear factor (NF)-kappaB pathway repressed cytokine-induced alterations in HS and gene expression of modifying enzymes. Firm adhesion of leukocytes to activated mouse glomerular endothelial cells decreased after removal of endothelial HS or addition of sulfated heparinoids. Specific antibodies that block N- and 6-O-sulfated HS domains on activated mouse endothelial cells reduced the number of rolling and firmly adhering leukocytes under dynamic flow conditions, while they increased the average leukocyte-rolling velocity. Our study shows that N- and 6-O-sulfated domains in HS on activated glomerular endothelium are crucial for leukocyte trafficking and are possible therapeutic targets. PMID- 17914354 TI - Adaptation to hypoxia in the diabetic rat kidney. AB - Hypoxia of the kidney in diabetes could predispose it to develop acute and chronic renal failure. To examine the relationship between renal hypoxia and renal failure, we measured hypoxia (as a pimonidazole adducts), hypoxia-inducible factors (HIFs), and a hypoxia target gene heme oxygenase-1. The studies were performed in rats with streptozotocin (STZ)-induced diabetes, Cohen diabetes sensitive rats, and during short-term artificial hyperglycemia in rats induced by intravenous glucose and octreotide. STZ-treated rats received insulin, the superoxide dismutase mimetic tempol, or contrast medium. Radiocontrast media causes hypoxia and HIF induction. Hypoxia, HIFs, and heme oxygenase were undetectable in controls, but transiently activated in STZ-treated and the Cohen diabetes sensitive rats. Different patterns of HIFs and pimonidazole were observed between the three models. Insulin abolished pimonidazole and HIF induction, whereas tempol lead to increased HIFs and heme oxygenase induction at similar levels of pimonidazole. When compared with control rats, STZ-treated rats exhibited more intense and protracted renal pimonidazole, with augmented hypoxia inducible factor production and reduced GFR following contrast media. Our data suggest that both regional hypoxia and hypoxia adaptation transiently occur in early stages of experimental diabetes, largely dependent on hyperglycemia or after contrast media. Tempol may augment the HIF response in diabetes. PMID- 17914355 TI - Regulation of cell cycle progression and gene expression by H2A deubiquitination. AB - Post-translational histone modifications have important regulatory roles in chromatin structure and function. One example of such modifications is histone ubiquitination, which occurs predominately on histone H2A and H2B. Although the recent identification of the ubiquitin ligase for histone H2A has revealed important roles for H2A ubiquitination in Hox gene silencing as well as in X chromosome inactivation, the enzyme(s) involved in H2A deubiquitination and the function of H2A deubiquitination are not known. Here we report the identification and functional characterization of the major deubiquitinase for histone H2A, Ubp M (also called USP16). Ubp-M prefers nucleosomal substrates in vitro, and specifically deubiquitinates histone H2A but not H2B in vitro and in vivo. Notably, knockdown of Ubp-M in HeLa cells results in slow cell growth rates owing to defects in the mitotic phase of the cell cycle. Further studies reveal that H2A deubiquitination by Ubp-M is a prerequisite for subsequent phosphorylation of Ser 10 of H3 and chromosome segregation when cells enter mitosis. Furthermore, we demonstrate that Ubp-M regulates Hox gene expression through H2A deubiquitination and that blocking the function of Ubp-M results in defective posterior development in Xenopus laevis. This study identifies the major deubiquitinase for histone H2A and demonstrates that H2A deubiquitination is critically involved in cell cycle progression and gene expression. PMID- 17914356 TI - A translocation signal for delivery of oomycete effector proteins into host plant cells. AB - Bacterial, oomycete and fungal plant pathogens establish disease by translocation of effector proteins into host cells, where they may directly manipulate host innate immunity. In bacteria, translocation is through the type III secretion system, but analogous processes for effector delivery are uncharacterized in fungi and oomycetes. Here we report functional analyses of two motifs, RXLR and EER, present in translocated oomycete effectors. We use the Phytophthora infestans RXLR-EER-containing protein Avr3a as a reporter for translocation because it triggers RXLR-EER-independent hypersensitive cell death following recognition within plant cells that contain the R3a resistance protein. We show that Avr3a, with or without RXLR-EER motifs, is secreted from P. infestans biotrophic structures called haustoria, demonstrating that these motifs are not required for targeting to haustoria or for secretion. However, following replacement of Avr3a RXLR-EER motifs with alanine residues, singly or in combination, or with residues KMIK-DDK--representing a change that conserves physicochemical properties of the protein--P. infestans fails to deliver Avr3a or an Avr3a-GUS fusion protein into plant cells, demonstrating that these motifs are required for translocation. We show that RXLR-EER-encoding genes are transcriptionally upregulated during infection. Bioinformatic analysis identifies 425 potential genes encoding secreted RXLR-EER class proteins in the P. infestans genome. Identification of this class of proteins provides unparalleled opportunities to determine how oomycetes manipulate hosts to establish infection. PMID- 17914357 TI - Neanderthals in central Asia and Siberia. AB - Morphological traits typical of Neanderthals began to appear in European hominids at least 400,000 years ago and about 150,000 years ago in western Asia. After their initial appearance, such traits increased in frequency and the extent to which they are expressed until they disappeared shortly after 30,000 years ago. However, because most fossil hominid remains are fragmentary, it can be difficult or impossible to determine unambiguously whether a fossil is of Neanderthal origin. This limits the ability to determine when and where Neanderthals lived. To determine how far to the east Neanderthals ranged, we determined mitochondrial DNA (mtDNA) sequences from hominid remains found in Uzbekistan and in the Altai region of southern Siberia. Here we show that the DNA sequences from these fossils fall within the European Neanderthal mtDNA variation. Thus, the geographic range of Neanderthals is likely to have extended at least 2,000 km further to the east than commonly assumed. PMID- 17914358 TI - Phagocyte-derived catecholamines enhance acute inflammatory injury. AB - It is becoming increasingly clear that the autonomic nervous system and the immune system demonstrate cross-talk during inflammation by means of sympathetic and parasympathetic pathways. We investigated whether phagocytes are capable of de novo production of catecholamines, suggesting an autocrine/paracrine self regulatory mechanism by catecholamines during inflammation, as has been described for lymphocytes. Here we show that exposure of phagocytes to lipopolysaccharide led to a release of catecholamines and an induction of catecholamine-generating and degrading enzymes, indicating the presence of the complete intracellular machinery for the generation, release and inactivation of catecholamines. To assess the importance of these findings in vivo, we chose two models of acute lung injury. Blockade of alpha2-adrenoreceptors or catecholamine-generating enzymes greatly suppressed lung inflammation, whereas the opposite was the case either for an alpha2-adrenoreceptor agonist or for inhibition of catecholamine degrading enzymes. We were able to exclude T cells or sympathetic nerve endings as sources of the injury-modulating catecholamines. Our studies identify phagocytes as a new source of catecholamines, which enhance the inflammatory response. PMID- 17914359 TI - Architectural and mechanistic insights into an EHD ATPase involved in membrane remodelling. PMID- 17914363 TI - Time for a fresh start. PMID- 17914364 TI - Living with the heat. PMID- 17914365 TI - On the paper trail. PMID- 17914369 TI - Ocean-drilling vessel should soon be afloat again. PMID- 17914368 TI - Sic is not enough; historical wording should be changed. PMID- 17914370 TI - Organic synthesis remains relevant in drug discovery. PMID- 17914371 TI - Biometrics: easy to steal, hard to regain identity. PMID- 17914372 TI - Biometrics: still much too unreliable for everyday use. PMID- 17914373 TI - Russian science: breaking up is hard to do. PMID- 17914379 TI - Russian science: the little ball made science bigger. PMID- 17914380 TI - How the mighty have fallen. PMID- 17914381 TI - Neuroscience: a local route to pain relief. PMID- 17914382 TI - Atomic physics: cold meeting at a junction. PMID- 17914384 TI - Development: inflationary pressures. PMID- 17914385 TI - Materials science: colloidal crystals find new order. PMID- 17914386 TI - Imaging technology: harmonic pictures in a flash. PMID- 17914388 TI - Chemical biology: More charges against aggregation. PMID- 17914389 TI - Mesenchymal stem cells within tumour stroma promote breast cancer metastasis. AB - Mesenchymal stem cells have been recently described to localize to breast carcinomas, where they integrate into the tumour-associated stroma. However, the involvement of mesenchymal stem cells (or their derivatives) in tumour pathophysiology has not been addressed. Here, we demonstrate that bone-marrow derived human mesenchymal stem cells, when mixed with otherwise weakly metastatic human breast carcinoma cells, cause the cancer cells to increase their metastatic potency greatly when this cell mixture is introduced into a subcutaneous site and allowed to form a tumour xenograft. The breast cancer cells stimulate de novo secretion of the chemokine CCL5 (also called RANTES) from mesenchymal stem cells, which then acts in a paracrine fashion on the cancer cells to enhance their motility, invasion and metastasis. This enhanced metastatic ability is reversible and is dependent on CCL5 signalling through the chemokine receptor CCR5. Collectively, these data demonstrate that the tumour microenvironment facilitates metastatic spread by eliciting reversible changes in the phenotype of cancer cells. PMID- 17914390 TI - Extremely fast acceleration of cosmic rays in a supernova remnant. AB - Galactic cosmic rays (CRs) are widely believed to be accelerated by shock waves associated with the expansion of supernova ejecta into the interstellar medium. A key issue in this long-standing conjecture is a theoretical prediction that the interstellar magnetic field can be substantially amplified at the shock of a young supernova remnant (SNR) through magnetohydrodynamic waves generated by cosmic rays. Here we report a discovery of the brightening and decay of X-ray hot spots in the shell of the SNR RX J1713.7-3946 on a one-year timescale. This rapid variability shows that the X-rays are produced by ultrarelativistic electrons through a synchrotron process and that electron acceleration does indeed take place in a strongly magnetized environment, indicating amplification of the magnetic field by a factor of more than 100. The X-ray variability also implies that we have witnessed the ongoing shock-acceleration of electrons in real time. Independently, broadband X-ray spectrometric measurements of RX J1713.7-3946 indicate that electron acceleration proceeds in the most effective ('Bohm diffusion') regime. Taken together, these two results provide a strong argument for acceleration of protons and nuclei to energies of 1 PeV (10(15) eV) and beyond in young supernova remnants. PMID- 17914391 TI - The a.c. and d.c. Josephson effects in a Bose-Einstein condensate. AB - The alternating- and direct-current (a.c. and d.c.) Josephson effects were first discovered in a system of two superconductors, the macroscopic wavefunctions of which are weakly coupled via a tunnelling barrier. In the a.c. Josephson effect, a constant chemical potential difference (voltage) is applied, which causes an oscillating current to flow through the barrier. Because the frequency is proportional to the chemical potential difference only, the a.c. Josephson effect serves as a voltage standard. In the d.c. Josephson effect, a small constant current is applied, resulting in a constant supercurrent flowing through the barrier. In a sense, the particles do not 'feel' the presence of the tall tunnelling barrier, and flow freely through it with no driving potential. Bose Einstein condensates should also support Josephson effects; however, while plasma oscillations have been seen in a single Bose-Einstein condensate Josephson junction, the a.c. Josephson effect remains elusive. Here we observe the a.c. and d.c. Josephson effects in a single Bose-Einstein condensate Josephson junction. The d.c. Josephson effect has been observed previously only in superconducting systems; in our study, it is evident when we measure the chemical potential current relation of the Bose-Einstein condensate Josephson junction. Our system constitutes a trapped-atom interferometer with continuous readout, which operates on the basis of the a.c. Josephson effect. In addition, the measured chemical potential-current relation shows that the device is suitable for use as an analogue of the superconducting quantum interference device, which would sense rotation. PMID- 17914392 TI - Fluctuating superconductivity in organic molecular metals close to the Mott transition. AB - On cooling through the transition temperature T(c) of a conventional superconductor, an energy gap develops as the normal-state charge carriers form Cooper pairs; these pairs form a phase-coherent condensate that exhibits the well known signatures of superconductivity: zero resistivity and the expulsion of magnetic flux (the Meissner effect). However, in many unconventional superconductors, the formation of the energy gap is not coincident with the formation of the phase-coherent superfluid. Instead, at temperatures above the critical temperature a range of unusual properties, collectively known as 'pseudogap phenomena', are observed. Here we argue that a key pseudogap phenomenon-fluctuating superconductivity occurring substantially above the transition temperature-could be induced by the proximity of a Mott-insulating state. The Mott-insulating state in the kappa-(BEDT-TTF)2X organic molecular metals can be tuned, without doping, through superconductivity into a normal metallic state as a function of the parameter t/U, where t is the tight-binding transfer integral characterizing the metallic bandwidth and U is the on-site Coulomb repulsion. By exploiting a particularly sensitive probe of superconducting fluctuations, the vortex-Nernst effect, we find that a fluctuating regime develops as t/U decreases and the role of Coulomb correlations increases. PMID- 17914393 TI - Single artificial-atom lasing. AB - Solid-state superconducting circuits are versatile systems in which quantum states can be engineered and controlled. Recent progress in this area has opened up exciting possibilities for exploring fundamental physics as well as applications in quantum information technology; in a series of experiments it was shown that such circuits can be exploited to generate quantum optical phenomena, by designing superconducting elements as artificial atoms that are coupled coherently to the photon field of a resonator. Here we demonstrate a lasing effect with a single artificial atom--a Josephson-junction charge qubit--embedded in a superconducting resonator. We make use of one of the properties of solid state artificial atoms, namely that they are strongly and controllably coupled to the resonator modes. The device is essentially different from existing lasers and masers; one and the same artificial atom excited by current injection produces many photons. PMID- 17914394 TI - Persistence of full glacial conditions in the central Pacific until 15,000 years ago. AB - The magnitude of atmospheric cooling during the Last Glacial Maximum and the timing of the transition into the current interglacial period remain poorly constrained in tropical regions, partly because of a lack of suitable climate records. Glacial moraines provide a method of reconstructing past temperatures, but they are relatively rare in the tropics. Here we present a reconstruction of atmospheric temperatures in the central Pacific during the last deglaciation on the basis of cosmogenic 3He ages of moraines and numerical modelling of the ice cap on Mauna Kea volcano, Hawaii--the only highland in the central Pacific on which moraines that formed during the last glacial period are preserved. Our reconstruction indicates that the Last Glacial Maximum occurred between 19,000 and 16,000 years ago in this region and that temperatures at high elevations were about 7 degrees C lower than today during this interval. Glacial retreat began about 16,000 years ago, but temperatures were still about 6.5 degrees C lower than today until 15,000 years ago. When combined with estimates of sea surface temperatures in the central Pacific Ocean, our reconstruction indicates that the lapse rate during the Last Glacial Maximum was higher than at present, which is consistent with the proposal that the atmosphere was drier at that time. Furthermore, the persistence of full glacial conditions until 15,000 years ago is consistent with the relatively late and abrupt transition to warmer temperatures in Greenland, indicating that there may have been an atmospheric teleconnection between the central Pacific and North Atlantic regions during the last deglaciation. PMID- 17914395 TI - An epipodite-bearing crown-group crustacean from the Lower Cambrian. AB - Crown-group crustaceans (Eucrustacea) are common in the fossil record of the past 500 million years back to the early Ordovician period, and very rare representatives are also known from the late Middle and Late Cambrian periods. Finds in Lower Cambrian rocks of the Phosphatocopina, the fossil sister group to eucrustaceans, imply that members of the eucrustacean stem lineage co-occurred, but it remained unclear whether crown-group members were also present at that time. 'Orsten'-type fossils are typically tiny embryos and cuticle-bearing animals, of which the cuticle is phosphatized and the material is three dimensional and complete with soft parts. Such fossils are found predominantly in the Cambrian and Ordovician and provide detailed morphological and phylogenetic information on the early evolution of metazoans. Here we report an Orsten-type Konservat-Lagerstatte from the Lower Cambrian of China that contains at least three new arthropod species, of which we describe the most abundant form on the basis of exceptionally well preserved material of several growth stages. The limb morphology and other details of this new species are markedly similar to those of living cephalocarids, branchiopods and copepods and it is assigned to the Eucrustacea, thus representing the first undoubted crown-group crustacean from the early Cambrian. Its stratigraphical position provides substantial support to the proposition that the main cladogenic event that gave rise to the Arthropoda was before the Cambrian. Small leaf-shaped structures on the outer limb base of the new species provide evidence on the long-debated issue of the origin of epipodites: they occur in a set of three, derive from setae and are a ground pattern feature of Eucrustacea. PMID- 17914396 TI - Reconciling complexity with stability in naturally assembling food webs. AB - Understanding how complex food webs assemble through time is fundamental both for ecological theory and for the development of sustainable strategies of ecosystem conservation and restoration. The build-up of complexity in communities is theoretically difficult, because in random-pattern models complexity leads to instability. There is growing evidence, however, that nonrandom patterns in the strengths of the interactions between predators and prey strongly enhance system stability. Here we show how such patterns explain stability in naturally assembling communities. We present two series of below-ground food webs along natural productivity gradients in vegetation successions. The complexity of the food webs increased along the gradients. The stability of the food webs was captured by measuring the weight of feedback loops of three interacting 'species' locked in omnivory. Low predator-prey biomass ratios in these omnivorous loops were shown to have a crucial role in preserving stability as productivity and complexity increased during succession. Our results show the build-up of food-web complexity in natural productivity gradients and pin down the feedback loops that govern the stability of whole webs. They show that it is the heaviest three-link feedback loop in a network of predator-prey effects that limits its stability. Because the weight of these feedback loops is kept relatively low by the biomass build-up in the successional process, complexity does not lead to instability. PMID- 17914397 TI - Inhibition of nociceptors by TRPV1-mediated entry of impermeant sodium channel blockers. AB - Most local anaesthetics used clinically are relatively hydrophobic molecules that gain access to their blocking site on the sodium channel by diffusing into or through the cell membrane. These anaesthetics block sodium channels and thereby the excitability of all neurons, not just sensory neurons. We tested the possibility of selectively blocking the excitability of primary sensory nociceptor (pain-sensing) neurons by introducing the charged, membrane-impermeant lidocaine derivative QX-314 through the pore of the noxious-heat-sensitive TRPV1 channel. Here we show that charged sodium-channel blockers can be targeted into nociceptors by the application of TRPV1 agonists to produce a pain-specific local anaesthesia. QX-314 applied externally had no effect on the activity of sodium channels in small sensory neurons when applied alone, but when applied in the presence of the TRPV1 agonist capsaicin, QX-314 blocked sodium channels and inhibited excitability. Inhibition by co-applied QX-314 and capsaicin was restricted to neurons expressing TRPV1. Injection of QX-314 together with capsaicin into rat hindpaws produced a long-lasting (more than 2 h) increase in mechanical and thermal nociceptive thresholds. Long-lasting decreases in pain sensitivity were also seen with regional injection of QX-314 and capsaicin near the sciatic nerve; however, in contrast to the effect of lidocaine, the application of QX-314 and capsaicin together was not accompanied by motor or tactile deficits. PMID- 17914399 TI - Genomics: the personal side of genomics. PMID- 17914405 TI - Oral administration of doxycycline allows tight control of transgene expression: a key step towards gene therapy of retinal diseases. AB - Gene transfer of neurotrophic or antiangiogenic factors has been shown to improve photoreceptor survival in retinal degenerative disorders (that is retinitis pigmentosa) and to prevent neovascularization in retinal vascular diseases (that is age-related macular degeneration, diabetic retinopathy). Expression of such neurotrophic or antiangiogenic factors after gene transfer requires the use of a regulatory system to control transgene expression to avoid unwanted side effects in cases of overexpression. In a previous study, we demonstrated that rAAV mediated gene transfer of the tetracycline-regulatable (tetR) system allows transgene regulation in the retina of nonhuman primates after intravenous administration of doxycycline (Dox). The purpose of this study was to evaluate oral administration of Dox to control transgene expression in the retina, since the pharmacokinetics after oral administration of the inducer drug represent a key factor when considering advancing to clinical trials. We report on the outcome of this evaluation and demonstrate that oral administration of Dox at a dose that is clinically used in humans (5 mg kg(-1) per day) is capable to continuously induce transgene expression in all macaques tested for 6 months. Moreover, control of transgene expression persists up to 4 years post-subretinal injection, with maximal induced levels of transgene product remaining stable over time. PMID- 17914404 TI - microRNAs join the p53 network--another piece in the tumour-suppression puzzle. AB - Several recent studies have found a conserved microRNA (miRNA) family, the miR 34s, to be direct transcriptional targets of p53. miR-34 activation can recapitulate elements of p53 activity, including induction of cell-cycle arrest and promotion of apoptosis, and loss of miR-34 can impair p53-mediated cell death. These data reinforce the growing awareness that non-coding RNAs are key players in tumour development by placing miRNAs in a central role in a well-known tumour-suppressor network. PMID- 17914406 TI - Protecting neurons from HIV-1 gp120-induced oxidant stress using both localized intracerebral and generalized intraventricular administration of antioxidant enzymes delivered by SV40-derived vectors. AB - Human immunodeficiency virus-1 (HIV-1) is the most frequent cause of dementia in adults under 40. We sought to use gene delivery to protect from HIV-1-related neuron loss. Because HIV-1 envelope (Env) gp120 elicits oxidant stress and apoptosis in cultured neurons, we established reproducible parameters of Env mediated neurotoxicity in vivo, then tested neuroprotection using gene delivery of antioxidant enzymes. We injected 100-500 ng mul(-1)gp120 stereotaxically into rat caudate-putamens (CP) and assayed brains for apoptosis by terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) 6-h to 14-day post-injection. Peak apoptosis occurred 1 day after injection of 250 and 500 ng microl(-1)gp120. TUNEL-positive cells mostly expressed neuronal markers (NeuroTrace), although some expressed CD68 and so were most likely microglial cells. Finally, we compared neuroprotection from gp120-induced apoptosis provided by localized and generalized intra-central nervous system (CNS) gene delivery. Recombinant SV40 vectors carrying Cu/Zn superoxide dismutase (SOD1) or glutathione peroxidase (GPx1) were injected into the CP, where gp120 was administered 4-24 weeks later. Alternatively, we inoculated the vector into the lateral ventricle (LV), with or without prior intraperitoneal (i.p.) administration of mannitol. Intracerebral injection of SV(SOD1) or SV(GPx1) significantly protected neurons from gp120-induced apoptosis throughout the 24 week study. Intraventricular vector administration protected from gp120 neurotoxicity comparably, particularly if preceded by mannitol i.p. Thus, HIV-1 gp120 is neurotoxic in vivo, and intracerebral or intra-ventricular administration of rSV40 vectors carrying antioxidant enzymes is neuroprotective. These findings suggest the potential utility of both localized and widespread gene delivery in treating neuroAIDS and other CNS diseases characterized by excessive oxidative stress. PMID- 17914407 TI - Recombinant Newcastle disease virus (NDV) with inserted gene coding for GM-CSF as a new vector for cancer immunogene therapy. AB - This is the first report describing recombinant (rec) Newcastle disease virus (NDV) as vector for gene therapy of cancer. The gene encoding granulocyte/macrophage colony-stimulating factor (GM-CSF) was inserted as an additional transcription unit at two different positions into the NDV genome. The rec virus with the strongest production of the gene product (rec(GM-CSF)) was selected for our study. The insertion of the new foreign gene did neither affect the main features of NDV replication nor its tumor selectivity. The gene product was biologically active and stable. Tumor vaccine cells infected by rec(GM-CSF) stimulated human peripheral blood mononuclear cells (PBMC) to exert antitumor bystander effects in vitro in a tumor neutralization assay. These effects were significantly increased when compared to vaccine infected by rec(-) virus. Furthermore, rec(GM-CSF) led to a much higher interferon-alpha (IFN-alpha) production than rec(-) when added as virus or as virus-modified vaccine to PBMC. Two distinct cell types, monocytes and plasmacytoid dendritic cells were shown to contribute to the augmented IFN-alpha response of PBMC. In conclusion, the already inherent anti-neoplastic and immunostimulatory properties of NDV could be further augmented by the introduction of a therapeutic gene whose product initiates a broad cascade of immunological effects in the microenvironment of the vaccine. PMID- 17914408 TI - The severity of FIP1L1-PDGFRA-positive chronic eosinophilic leukaemia is associated with polymorphic variation at the IL5RA locus. AB - We have investigated the hypothesis that constitutional genetic variation in IL-5 signalling may be associated with the development or severity of FIP1L1-PDGFRA positive chronic eosinophilic leukaemia (CEL) in humans. We genotyped six single nucleotide polymorphisms (SNP) within or close to the IL5RA or IL5 genes in 82 patients with FIP1L1-PDGFRA-positive CEL plus, as controls, healthy individuals (n=100), patients with FIP1L1-PDGFRA-negative eosinophilia (n=100) or patients with chronic myeloid leukaemia (CML) (n=100). We found no association between SNP allele frequency between FIP1L1-PDGFRA-positive and control cases. However, for FIP1L1-PDGFRA cases, we found an association between the genotype at rs4054760, an SNP in the 5'-UTR of IL5RA and peripheral blood eosinophil count (P=0.026) as well as the presence or absence of tissue infiltration (P=0.032). Although these associations fell below the level of significance once corrected for multiple testing, no such association was seen in FIP1L1-PDGFRA-negative cases and no difference in allele frequencies for rs4054760 was seen in control populations across Europe. Furthermore, in an analysis of 112 patients with CML, IL5RA expression was strongly related to rs4054760 genotype (P<0.001). These data suggest that the variations in IL5RA expression are linked to constitutional IL5RA genotype and severity of FIP1L1-PDGFRA disease. PMID- 17914409 TI - Contribution of GM-CSF and IL-8 to the CD44-induced differentiation of acute monoblastic leukemia. PMID- 17914410 TI - Genomic analysis of different clonal evolution in a twin pair with t(12;21) positive acute lymphoblastic leukemia sharing the same prenatal clone. PMID- 17914411 TI - Two novel JAK2 exon 12 mutations in JAK2V617F-negative polycythaemia vera patients. PMID- 17914412 TI - Lymphoma microenvironment: culprit or innocent? AB - Studies are revealing that lymphoid neoplasms are characterized by well-defined chromosome translocations and by the accumulation of subsequent molecular alterations involving mainly the cell cycle and/or apoptotic pathways. However, survival of B and T tumor cells is also dependent on the interactions with the accompanying cells that comprise the lymphoma microenvironment. Although non tumor cells can contribute both positive and negative signals to the lymphoma cells, in this review we present compelling evidence of the essential influence of the tumor microenvironment on the initiation and progression of specific lymphoma types, highlighting some new therapeutic approaches that target the lymphoma microenvironment. PMID- 17914413 TI - Determination of IGHV gene mutational status in chronic lymphocytic leukemia: bioinformatics advances meet clinical needs. PMID- 17914415 TI - HOpe for contrast-induced acute kidney injury. AB - Contrast-induced nephropathy (CIN) is a common cause of acute kidney injury in hospitalized patients. The mechanisms involved in the pathogenesis of CIN are incompletely understood. Goodman et al. have demonstrated for the first time that heme oxygenase-1, a 32-kilodalton protein with antioxidant, antiapoptotic, anti inflammatory effects, is induced in the kidney and, importantly, provides a beneficial effect in CIN. PMID- 17914416 TI - Vitamin D as a novel nontraditional risk factor for mortality in hemodialysis patients: the need for randomized trials. AB - Vitamin D has been used in the context of secondary hyperparathyroidism in patients with end-stage renal disease. A wave of recent studies suggests that vitamin D treatment may be associated with decreased mortality risk in these patients. The article by Wolf et al. further supports these studies by identifying vitamin D deficiency as a risk factor for early mortality in incident hemodialysis patients. PMID- 17914417 TI - A cost too high to bear? Prophylaxis versus preemptive therapy to prevent post transplantation cytomegalovirus. AB - Both prophylaxis and preemptive therapy are used to prevent the development of cytomegalovirus (CMV) disease after transplantation. Preemptive therapy exposes the least number of patients to costly and potentially toxic drugs. Prophylaxis is less labor intensive and requires less expensive monitoring. While the overall cost of the two modalities is similar, current literature suggests that prophylaxis has an advantage in avoiding secondary effects of CMV. Randomized comparative trials are imperative. PMID- 17914418 TI - Collecting duct epithelium and injury: not all cells are created equal. AB - Studies by Butt et al. in the developing fetus provide new and timely insights into the regulation of repair and fibrosis in the injured kidney. Using a clinically relevant model, they have examined the response of the medullary collecting duct to ureteral obstruction, with some unexpected findings. PMID- 17914422 TI - Make your diagnosis. Multiple myeloma-associated with spurious hyperphosphatemia. PMID- 17914423 TI - Microscopic hematuria and the posterior nutcracker phenomenon. PMID- 17914424 TI - Steroid-induced striae in severe nephrotic syndrome. PMID- 17914426 TI - Childhood blindness: a UK epidemiological perspective. AB - The purposes of this short paper are to introduce the reader to the aims of ophthalmic epidemiology and its relevance to clinical practice in ophthalmology and to vision science research. Summarise what is known currently about the epidemiology of severe visual impairment and blindness in childhood in the UK. Highlight the key gaps in our knowledge in this area and identify some of the most pressing challenges in terms of research. PMID- 17914427 TI - Ophthalmological problems associated with preterm birth. AB - As survival of preterm infants improves, the long-term care of consequent ophthalmic problems is an expanding field. Preterm birth can inflict a host of challenges on the developing ocular system, resulting in the visual manifestations of varied significance and pathological scope. The ophthalmic condition most commonly associated with preterm birth is retinopathy of prematurity, which has the potential to result in devastating vision loss. However, the visual compromise from increased incidence of refractive errors, strabismus, and cerebral vision impairment has significant impact on visual function, which also has influence on other developmental aspects including psychological and educational. In this review, the normal ocular development is discussed, aiming to exemplify the impact of early exteriorisation on one of the more naive organs of prematurity. This is then related to the incidence and visual consequences of many types of deficit, including refractive error, strabismus, and loss of visual function in preterm populations, with comparisons to term infant studies. Often these conditions are linked with causal and resultant factors being impossible to segregate, but the common factor of increased rates of all types of ophthalmic deficits demonstrates that children born prematurely are indeed premature for life. PMID- 17914428 TI - Mechanisms of brain injury in the newborn. AB - Advances in obstetric and neonatal medical care have led to marked improvements in the chances of survival for extremely preterm and low birth weight babies. This review focuses on the mechanisms of neurological injury in extremely preterm and critically ill infants and discusses current progress in therapeutic strategies. PMID- 17914429 TI - Chiasm formation in man is fundamentally different from that in the mouse. AB - At the optic chiasm axons make a key binary decision either to cross the chiasmal midline to innervate the contralateral optic tract or to remain uncrossed and innervate the ipsilateral optic tract. In rodents, midline interactions between axons from the two eyes are critical for normal chiasm development. When one eye is removed early in development the hemispheric projections from the remaining eye are disrupted, increasing the crossed projection at the expense of the uncrossed. This is similar to the abnormal decussation pattern seen in albinos. The decussation pattern in marsupials, however, is markedly different. Early eye removal in the marsupial has no impact on projections from the remaining eye. These differences are related to the location of the uncrossed projection through the chiasm. In rodents, axons that will form the uncrossed projection approach the chiasmal midline, while in marsupials they remain segregated laterally through the chiasm. Histological analysis of the optic chiasm in man provides anatomical evidence to suggest that, unlike in rodents, uncrossed axons are confined laterally from the optic nerve through to the optic tract and do not mix in each hemi-chiasm. This is a pattern similar to that found in marsupials. Electrophysiological evidence in human anophthalmics shows that the failure of one eye to develop in man has no impact on the hemispheric projections from the remaining eye. This strongly suggests that the mechanisms regulating chiasmal development in man differ from those in rodents, but may be similar to marsupials. This implies that optic chiasm formation in rodents and ferrets is not common to placental mammals in general. PMID- 17914430 TI - Developmental abnormalities of the optic nerve and chiasm. AB - Developmental anomalies of the optic nerve are an important and growing cause of lifelong visual handicap and they are often associated with systemic abnormalities. This review focuses on the ocular and systemic aspects of developmental anomalies arising from defects of fetal fissure closure and retinal ganglion cell development, and covers some other optic-disc anomalies that have systemic significance. PMID- 17914431 TI - Brain injury and the eye. AB - Visual loss associated with brain damage is the single greatest cause of visual impairment in children in developed countries. Damage may occur in any of five separate visual systems: primary visual cortex, visual associative cortex area, optic radiations, optic nerves, and visual attention pathways. Improving our understanding of the pathophysiology of these causes for visual loss may lead to better rehabilitation and educational strategies for these children. PMID- 17914432 TI - A practical guide to the management of anophthalmia and microphthalmia. AB - Congenital anophthalmia and microphthalmia are rare developmental defects of the globe. They often arise in conjunction with other ocular defects such as coloboma and orbital cyst. They may also be part of more generalised syndromes, such as CHARGE syndrome. Anophthalmia, microphthalmia, and coloboma are likely to be caused by disturbances of the morphogenetic pathway that controls eye development, either as a result of primary genetic defect, or external gestational factors, including infection or drugs that can influence the smooth processes of morphogenesis. The ophthalmologist is often the primary carer for children with anophthalmia and microphthalmia, and as such can coordinate the multidisciplinary input needed to offer optimal care for these individuals, including vision and family support services. They are able to assess the vision and maximise the visual potential of the child and they can also ensure that the cosmetic and social impact of anophthalmia or microphthalmia is minimised by starting socket expansion or referring to a specialist oculoplastics and prosthetics unit. A coordinated approach with paediatrics is necessary to manage any associated conditions. Genetic diagnosis and investigations can greatly assist in providing a diagnosis and informed genetic counselling. PMID- 17914433 TI - Advances in the management of congenital and infantile cataract. AB - Congenital and infantile cataracts produce deprivation amblyopia and can thus cause lifelong visual impairment. Successful management is dependent on early diagnosis and referral for surgery when indicated. Accurate optical rehabilitation and postoperative supervision are essential.The timing of surgery and its relationship to the duration of deprivation is important. Unilateral congenital cataract surgery within 6 weeks of birth produces the best outcomes. The equivalent 'latent' period for bilateral visual deprivation may be longer at around 10 weeks. Visual deprivation has a significant impact on the development of fixation stability. Major form deprivation, even after early surgery, leads to nystagmus. This is mostly manifest latent nystagmus (MLN). The latent period for fixation stability may be as short as 3 weeks. Preoperative congenital nystagmus (CN) can convert to more benign MLN after surgery. Infantile IOL implantation is becoming increasingly accepted. A satisfactory long-term refractive result requires that allowance be made for childhood axial growth and myopic shift. In a series of 25 infants (33 eyes) implanted before 12 months of age, the mean myopic shift at 12 months was 4.83 D. This increased to 5.3 D in infants implanted before 10 weeks. The initial desired refractive outcome following IOL implantation is thus hypermetropia, with the degree dependent on the age of the child. Glaucoma or ocular hypertension is a common complication following paediatric cataract surgery. Microphthalmia and surgery in early infancy are risk factors. Tonometry results may be influenced by the increased corneal thickness seen in aphakic and pseudophakic children. The long-term prognosis of eyes with aphakic glaucoma is not necessarily poor but intraocular pressure control may require three or more medications. Surgical intervention appears to be necessary in over a quarter of eyes. Posterior capsule opacification (PCO) is common in infants undergoing primary lens implantation. Primary capsulotomy and anterior vitrectomy reduce the risk of PCO. In the absence of anterior vitrectomy, primary posterior capsulotomy does not prevent visual axis opacification. Further developments will continue to be driven by clinical research. The prevention of capsule opacification and cellular proliferation may in future be achieved by the use of devices to specifically target epithelial cells at surgery. PMID- 17914434 TI - Molecular and developmental mechanisms of anterior segment dysgenesis. AB - Anterior segment dysgenesis (ASD) is a failure of the normal development of the tissues of the anterior segment of the eye. It leads to anomalies in the structure of the mature anterior segment, associated with an increased risk of glaucoma and corneal opacity. Several different gene mutations have been identified underlying these anomalies with the majority of ASD genes encoding transcriptional regulators. In this review, the role of the ASD genes, PITX2 and FOXC1, is considered in relation to the embryology of the anterior segment, the biochemical function of these proteins, and their role in development and disease aetiology. The emerging view is that these genes act in concert to specify a population of mesenchymal progenitor cells, mainly of neural crest origin, as they migrate anteriorly around the embryonic optic cup. These same genes then regulate mesenchymal cell differentiation to give rise to distinct anterior segment tissues. Development appears critically sensitive to gene dosage, and variation in the normal level of transcription factor activity causes a range of anterior segment anomalies. Interplay between PITX2 and FOXC1 in the development of different anterior segment tissues may partly explain the phenotypic variability and the genetic heterogeneity characteristic of ASD. PMID- 17914435 TI - Advances in the management of paediatric glaucoma. AB - The paediatric glaucomas present some of the greatest clinical challenges. We review the advances in the management of the paediatric glaucomas, which have improved the outlook for these patients and their families. These advances include improvements in diagnosis, investigations, anaesthetic techniques, medical, surgical, and laser therapies. PMID- 17914436 TI - Congenital corneal opacities - a surgical approach to nomenclature and classification. AB - The classification system of congenital corneal opacification (CCO) may be better considered from a perspective of pathogenesis, surgical intervention, and prognosis. The author feels that CCO is best considered as being primary and secondary. Primary CCO includes corneal dystrophies and choristomas presenting at birth. Secondary CCO may be best considered as cases of kerato-irido-lenticular dysgenesis (KILD) and other secondary causes including infection, iatrogenic, developmental anomalies of the iridotrabecular system or lens or both, and developmental anomalies of the adnexal. The appropriate classification may help determine prognosis of any surgical intervention. Terminology is crucial to furthering our understanding of the formation of the anterior chamber if we are to do so by studying cases of CCO. Peters' anomaly is too imprecise a term to describe cases of CCO. This classification of primary and secondary CCO with its subclassifications cannot be made by clinical examination alone and necessitates other diagnostic assessments. It is time to only accept studies of CCO genotype/phenotype correlation for publication if there is clinical phenotype augmented by anterior segment imaging (OCT or high-frequency ultrasound) or histology or both. PMID- 17914437 TI - Changing challenges in the control of blindness in children. AB - One cannot address issues of control of blindness in children without first considering the epidemiology, as this allows priorities to be determined and appropriate strategies to be delineated. Control does not occur within a vacuum, and so it is also important to understand the context in which programmes are to be implemented, particularly in relation to development and poverty--the distal risk factors that are powerful determinants of the incidence of disease in populations. Interventions need to be considered from the perspective of patients as well as providers if services are to be acceptable and appropriate. PMID- 17914438 TI - Molecular genetics of infantile-onset retinal dystrophies. AB - Over the last decade there have been major advances in our understanding of the molecular pathology of inherited retinal dystrophies. This paper reviews recent advances in the identification of genetic mutations underlying infantile-onset inherited retinal disorders and considers how this knowledge may lead to novel therapeutic approaches. PMID- 17914439 TI - Stem cell therapy and the retina. AB - Retinal degeneration culminating in photoreceptor loss is the leading cause of untreatable blindness in the developed world. In this review, we consider how photoreceptors might be replaced by transplantation and how stem cells might be optimised for use as donor cells in future clinical strategies for retinal repair. We discuss the current advances in human and animal models of retinal cell transplantation, focussing on stem cell and reproductive cloning biology, in relation to the practical issues of retinal transplantation surgery. Stem and progenitor cells can be isolated from a number of sources including embryonic tissue, adult brain and even the retina, prompting many researchers to investigate the potential for using these cells to generate photoreceptors for transplantation. Nevertheless, several obstacles need to be overcome before these techniques can be applied in a clinical setting. Embryonic or stem cells have so far shown little ability to differentiate into retinal phenotypes when transplanted into the adult retina. We have recently noted, however, that donor cells harvested much later, at the photoreceptor precursor developmental stage, can be transplanted successfully and restore visual function. The current challenge is to understand the developmental processes that guide embryonic or adult stem cells towards photoreceptor differentiation, so that large numbers of these cells might be transplanted at the optimal stage. Future advances in reproductive cloning technology could lead to the successful generation of stem cells from adult somatic cells, thereby facilitating auto-transplantation of genetically identical cells in patients requiring photoreceptor replacement. PMID- 17914440 TI - Vaccine immunopotentiators of the future. AB - Vaccine adjuvants or immunopotentiators comprise a diverse group of molecules or formulations. Despite a wealth of different candidates, there is a need for better vaccine adjuvants in both veterinary and human medicine. For human vaccines, the immunopotentiator choice has been limited to aluminum salts, until recently. By contrast, a whole range of adjuvants is employed for inactivated veterinary vaccines, due to less stringent safety and regulatory requirements and proven superior vaccine performance. This review highlights recent developments and future trends in immunopotentiators. PMID- 17914441 TI - Effect of semen sampling frequency on seminal antiretroviral drug concentration. AB - Study of male genital tract (MGT) pharmacology is relevant to the treatment of prostatitis, prostate cancer, infertility, and seminal human immunodeficiency virus transmission. However, the time course of drug concentrations in the MGT is largely unknown. To determine the feasibility of frequent semen sampling in assessing the pharmacokinetics of the MGT, we administered efavirenz, indinavir, and zidovudine to subjects to achieve steady-state levels and then collected semen samples at sequentially decreasing ejaculation intervals. The volume of seminal plasma decreased from 4.0 (1.2-5.1) ml (median with range) at 48 h after the baseline ejaculation to 0.72 (0.45-1.6) ml 1 h after a previous ejaculation, which was still adequate for drug concentration assessment. The seminal fructose concentration also decreased. However, the concentration of prostate-specific antigen and all three drugs did not decrease, even if the ejaculation intervals decreased to 1 h. Thus, semi-intensive semen sampling can be used to assess MGT pharmacokinetics. PMID- 17914442 TI - Population pharmacokinetics of melphalan and glutathione S-transferase polymorphisms in relation to side effects. AB - Melphalan is associated with severe side effects such as mucositis, diarrhea, and myelosuppression. We investigated how much the individual severity of these side effects is predicted by pharmacokinetics. In addition, we studied glutathione S transferase GSTM1, GSTT1, and GSTP1 polymorphisms in relation to adverse events. A high interindividual pharmacokinetic variability was observed in 84 patients. There was a linear correlation between creatinine and melphalan clearance (P=0.0004). Patients treated with a dose > or = 70 mg/m(2) had a 23-fold increased risk to develop mucositis (P<0.001) and a 12-fold increased risk to develop diarrhea (P<0.001) compared with lower doses. The GSTP1 codon 105 polymorphism may be relevant for development of mucositis and the GSTT1 deletion may predict diarrhea, but these findings require confirmation. Melphalan-induced side effects were significantly dependent only on dose. Therapeutic drug monitoring or genotyping for GST does not appear to be very helpful in optimizing therapy with melphalan. PMID- 17914443 TI - Uninteresting conflicts of interest. PMID- 17914444 TI - Topically applied nitric oxide induces T-lymphocyte infiltration in human skin, but minimal inflammation. AB - Nitric oxide (NO) plays an important role in the cutaneous response to UV radiation and in cutaneous inflammation. The presence of inducible NO synthase protein in a number of inflammatory dermatoses, coupled with the induction of an intense cutaneous inflammatory infiltrate following topical application of the NO donor-acidified nitrite (NO2(-)), has set the paradigm of NO being an inflammatory mediator in human skin. Using zeolite NO (Ze-NO), a chemically inert, pure NO donor, we have shown that NO per se produces little inflammation. Biologically, relevant doses of Ze-NO induce a dermal CD4-positive T-cell infiltrate and IFN-gamma secretion. In contrast acidified nitrite, releasing equal quantities of NO (measured by dermal microdialysis and cutaneous erythema), induces an intense epidermal infiltrate of macrophages with a similar dermal infiltrate of CD3-, CD4-, CD8-, and CD68-positive cells and neutrophils. Suction blisters were created in Ze-NO-treated and control skin. IFN-gamma, but not IL-4, was detected in Ze-NO-treated skin (mean control 0.1+/-0.07 pg mg(-1) protein, mean IFN-gamma 0.6+/-0.4 pg mg(-1) protein). We suggest that the potent inflammation induced by acidified NO2(-) is secondary to the release of additional mediators. PMID- 17914445 TI - Somatic mutation analysis in NF1 cafe au lait spots reveals two NF1 hits in the melanocytes. PMID- 17914447 TI - Mutations in the serum/glucocorticoid regulated kinase 3 (Sgk3) are responsible for the mouse fuzzy (fz) hair phenotype. PMID- 17914446 TI - Chronic diseases requiring hospitalization and risk of non-melanoma skin cancers- a population based study from Denmark. AB - We examined the associations between chronic diseases requiring hospitalization and the risk of non-melanoma skin cancers (NMSCs) in a population-based case control study of 4,187 patients diagnosed with a first primary NMSC in 1995 in Denmark. From the National Patient Registry covering all Danish hospitals, we obtained data on hospitalizations with chronic diseases, recorded before the date of NMSC diagnosis. Using incidence density sampling, we selected 10 age-, gender , and residence-matched controls from the Danish Civil Registration System. We used conditional logistic regression to compute incidence rate ratios (IRRs) and 95% confidence intervals (CIs). Although no overall association was found between basal cell carcinoma (BCC) and hospitalization for chronic diseases, an elevated IRR for BCC was found among patients with connective tissue disease (IRR 1.34 (95% CI: 0.99-1.82)), transplants (IRR 8.00 (95% CI: 2.15-30)), and lymphoma (IRR 2.50 (95% CI: 1.29-4.84)). An overall association between squamous cell carcinoma (SCC) and hospitalization for chronic diseases was found and specific among patients with leukemia (IRR 7.75 (95% CI: 2.35-26)), lymphoma (IRR 3.86 (95% CI: 0.99-15)), and skin diseases (IRR 5.28 (95% CI: 1.95-14)). Our study supports the presence of an association between certain chronic diseases and NMSC, and further suggests that these results unlikely are due to bias. JID JOURNAL CLUB ARTICLE: For questions, answers, and open discussion about this article please go to http://network.nature.com/group/jidclub. PMID- 17914448 TI - RSPO4 is the major gene in autosomal-recessive anonychia and mutations cluster in the furin-like cysteine-rich domains of the Wnt signaling ligand R-spondin 4. AB - Congenital anonychia is a rare autosomal-recessive disorder characterized by the absence of finger- and toenails. Recently, we and others identified the secreted Wnt signaling ligand R-spondin 4 (RSPO4) as the first gene known to be responsible for inherited anonychia. R-spondins are secreted proteins that activate the Wnt/beta-catenin signaling pathway. This puts anonychia on the growing list of congenital malformation syndromes caused by Wnt signaling pathway defects. Here, we expand the RSPO4 mutational spectrum by identification of the previously unknown mutations c.190C>T (p.Arg64Cys) in exon 2 and c.301C>T (p.Gln101X) in exon 3, thereby corroborating R-spondin 4 as the major protein in autosomal-recessive anonychia. Almost all RSPO4 mutations detected so far affect the highly conserved exons 2 and 3. Thus, we postulate that RSPO4 mutations preferentially cluster in the furin-like cysteine-rich domains of R-spondin 4, which is in line with experimental data proposing that for beta-catenin stabilization, a shortened protein comprising just these two regions is sufficient. PMID- 17914449 TI - Further exploring the brain-skin connection: stress worsens dermatitis via substance P-dependent neurogenic inflammation in mice. AB - A neurogenic component in atopy and allergy is evident and potentially of great pathogenic relevance. Stress was recently shown to activate elements of this component and is vividly discussed as a cause of exacerbation. However, to date, scientific proof of stress-induced neuronal plasticity and neuro-immune interaction in atopy or allergy remains lacking. Here we show early evidence that exposure to sound stress and atopic dermatitis-like allergic dermatitis (AD) equipotently raise the number of cutaneous nerve fibers containing the prototypic stress neuropeptide substance P (SP) in mice. Stress increases AD readout parameters by at least 30% (eosinophil infiltration, vascular cell adhesion molecule-positive blood vessels, epidermal thickness). This dramatic pathologic exacerbation is associated with increased neurogenic inflammation (degranulated mast cells; interstitial neuropeptidergic dense core granules, mast cell apoptosis, endothelial gaping). Key features of AD exacerbation could not be induced in mice lacking the neurokinin-1 SP receptor (NK1). Interestingly, stress had no significant additional effect on CD4+ cell number, but shifted the cytokine profile toward TH2 in skin. Thus, we conclude that stress primarily exacerbates AD via SP-dependent cutaneous neurogenic inflammation and subsequent local cytokine shifting and should be considered as a therapeutic target, while it offers a convincing pathogenic explanation to affected patients and their frustrated physicians alike. PMID- 17914450 TI - mTOR is activated in the majority of malignant melanomas. AB - The objective of this study was to determine whether activation of the kinase mammalian target of rapamycin (mTOR) is associated with human melanoma. We found moderate or strong hyperphosphorylation of ribosomal protein S6 in 78/107 melanomas (73%). In contrast, only 3/67 benign nevi (4%) were moderately positive, and none were strongly positive. These data indicate that mTOR activation is very strongly associated with malignant, compared to benign, melanocytic lesions. Next, we tested six melanoma-derived cell lines for evidence of mTOR dysregulation. Five of the six lines showed persistent phosphorylation of S6 after 18 hours of serum deprivation, and four had S6 phosphorylation after 30 minutes of amino-acid withdrawal, indicating inappropriate mTOR activation. The proliferation of three melanoma-derived lines was blocked by the mTOR inhibitor rapamycin, indicating that mTOR activation is a growth-promoting factor in melanoma-derived cells. mTOR is directly activated by the small guanosine triphosphatase Ras homolog enriched in brain (Rheb), in a farnesylation-dependent manner. Therefore, to investigate the mechanism of mTOR activation, we used the farnesyl transferase inhibitor FTI-277, which partially blocked the growth of three of the six melanoma cell lines. Together, these data implicate activation of mTOR in the pathogenesis of melanoma, and suggest that Rheb and mTOR may be targets for melanoma therapy. PMID- 17914451 TI - Activation of T-cells from allergic patients and volunteers by p-phenylenediamine and Bandrowski's base. AB - Allergic contact dermatitis is commonly associated with exposure to p phenylenediamine. The aim of this study was to determine whether p phenylenediamine (PPD) and/or Bandrowski's base (BB) stimulate T cells from allergic patients and volunteers, and to explore the relationship between T-cell immunogenicity and allergy. Lymphocytes from allergic patients proliferated with PPD and BB (n=8). Lymphocytes from 14/16 non-allergic individuals also proliferated following stimulation, but only with BB; cord blood lymphocytes failed to respond (n=6). Glutathione, which prevented BB formation, but not binding of PPD to cells and serum, did not prevent p-phenylenediamine-specific stimulation of patient lymphocytes. T-cell clones generated from allergic patients were stimulated separately with PPD and BB, while clones from volunteers proliferated with BB alone. Patient and volunteer clones secreted IL-4, IL-5, IL 13, TNF-alpha, MIP-1alpha, MIP-1beta, and RANTES. These data show that activation of T lymphocytes from allergic individuals alone with PPD represents an important discrimination between allergic and non-allergic groups. BB-specific T cells are found in both allergic patients and volunteers, but not in cord blood. Their presence seems to reflect an acquired immune response, which is not translated into an allergic reaction. PMID- 17914452 TI - Regulation of epidermal tight-junctions (TJ) during infection with exfoliative toxin-negative Staphylococcus strains. AB - Tight Junction (TJ) proteins have been shown to exert a barrier function within the skin. Here, we study the fate of TJ proteins during the challenge of the skin by bacterial colonization and infection. We investigated the influence of various exfoliative toxin-negative Staphylococcus strains on TJ, adherens junction (AJ), desmosomal proteins, and actin in a human keratinocyte infection culture and in a porcine skin infection model. We found that the pathogen Staphylococcus aureus downregulates TJ and subsequently AJ and desmosomal proteins, including atypical protein kinase C, an essential player in TJ formation, at the cell-cell borders of keratinocytes in a time and concentration dependent manner. Little changes in protein and RNA levels were seen, indicating redistribution of proteins. In cultured keratinocytes, a reduction of transepithelial resistance was observed. Staphylococcus epidermidis shows only minor effects. All strains induced enhanced expression of occludin and ZO-1 at the beginning of colonization/infection. Thus, we demonstrate that TJ are likely to be involved in skin infection of exfoliative toxin-negative S. aureus. As we did not find a change in actin, and as changes of TJ preceded alterations of AJs and desmosomes, we suggest that S. aureus targets TJ. PMID- 17914453 TI - The localization of label-retaining cells in mouse nails. PMID- 17914454 TI - Single-nucleotide-specific siRNA targeting in a dominant-negative skin model. AB - RNA interference offers a novel approach for developing therapeutics for dominant negative genetic disorders. The ability to inhibit expression of the mutant allele without affecting wild-type gene expression could be a powerful new treatment option. Targeting the single-nucleotide keratin 6a (K6a) N171K mutation responsible for the rare monogenic skin disorder pachyonychia congenita (PC), we demonstrate that small interfering RNAs (siRNAs) can potently and selectively block expression of mutant K6a. To test whether lead siRNAs could discriminate mutant mRNA in the presence of both wild-type and mutant forms, a dominant negative PC cell culture model was developed. As predicted for a dominant negative disease, simultaneous expression of both wild-type and mutant K6a resulted in defective keratin filament formation. Addition of mutant-specific siRNAs allowed normal filament formation, suggesting selective inhibition of mutant K6a. The effectiveness of our siRNA in skin was tested by co-delivering a firefly luciferase/mutant K6a bicistronic reporter construct and mutant-specific siRNAs to mouse footpads. Potent inhibition of the fluorescent reporter was demonstrated using the Xenogen IVIS200 in vivo imaging system. Additionally, wild type-specific siRNAs knocked down the expression of pre-existing endogenous K6a in human keratinocytes. These results suggest that efficient delivery of these "designer siRNAs" may allow effective treatment of numerous genetic disorders including PC. PMID- 17914455 TI - Shb gene knockdown increases the susceptibility of SVR endothelial tumor cells to apoptotic stimuli in vitro and in vivo. AB - The Shb adapter protein is an Src homology 2-domain containing signaling intermediate operating downstream of several tyrosine kinase receptors, including vascular endothelial growth factor receptor-2. Shb is multifunctional and apoptosis is one response that Shb regulates. Inhibition of angiogenesis can be used in cancer therapy, and one way to achieve this is by inducing endothelial cell apoptosis. The angiosarcoma cell line SVR is of endothelial origin and can be used as a tool for studying in vivo inhibition of angiogenesis, and we thus employed an Shb-knockdown strategy using an inducible lentiviral system to reduce Shb levels in SVR cells and to study their responses. Shb knockdown increases the susceptibility of SVR cells to the apoptotic agents, cisplatin and staurosporine. Simultaneously, Shb knockdown causes reduced focal adhesion kinase (FAK) activation, monitored as phosphorylation of the regulatory residues tyrosines 576/577. No detectable effects on Akt or extracellular signal-regulated kinase activity were noted. The altered FAK activity coincided with an elongated cell phenotype that was particularly noticeable in the presence of staurosporine. In order to relate the effects of Shb knockdown to in vivo tumorigenicity, cells were exposed to the angiogenesis inhibitor honokiol, and again the cells with reduced Shb content exhibited increased apoptosis. Tumor growth in vivo was strongly reduced in the Shb-knockdown cells upon honokiol treatment. It is concluded that Shb regulates apoptosis and cell shape in tumor endothelial cells via FAK, and that Shb is a potential target for inhibition of angiogenesis. PMID- 17914457 TI - Phosphorylation of Bem2p and Bem3p may contribute to local activation of Cdc42p at bud emergence. AB - Site-specific activation of the Rho-type GTPase Cdc42p is critical for the establishment of cell polarity. Here we investigated the role and regulation of the GTPase-activating enzymes (GAPs) Bem2p and Bem3p for Cdc42p activation and actin polarization at bud emergence in Saccharomyces cerevisiae. Bem2p and Bem3p are localized throughout the cytoplasm and the cell cortex in unbudded G1 cells, but accumulate at sites of polarization after bud emergence. Inactivation of Bem2p results in hyperactivation of Cdc42p and polarization toward multiple sites. Bem2p and Bem3p are hyperphosphorylated at bud emergence most likely by the Cdc28p-Cln2p kinase. This phosphorylation appears to inhibit their GAP activity in vivo, as non-phosphorylatable Bem3p mutants are hyperactive and interfere with Cdc42p activation. Taken together, our results indicate that Bem2p and Bem3p may function as global inhibitors of Cdc42p activation during G1, and their inactivation by the Cdc28p/Cln kinase contributes to site-specific activation of Cdc42p at bud emergence. PMID- 17914456 TI - Structural basis for the recruitment of profilin-actin complexes during filament elongation by Ena/VASP. AB - Cells sustain high rates of actin filament elongation by maintaining a large pool of actin monomers above the critical concentration for polymerization. Profilin actin complexes constitute the largest fraction of polymerization-competent actin monomers. Filament elongation factors such as Ena/VASP and formin catalyze the transition of profilin-actin from the cellular pool onto the barbed end of growing filaments. The molecular bases of this process are poorly understood. Here we present structural and energetic evidence for two consecutive steps of the elongation mechanism: the recruitment of profilin-actin by the last poly-Pro segment of vasodilator-stimulated phosphoprotein (VASP) and the binding of profilin-actin simultaneously to this poly-Pro and to the G-actin-binding (GAB) domain of VASP. The actin monomer bound at the GAB domain is proposed to be in position to join the barbed end of the growing filament concurrently with the release of profilin. PMID- 17914458 TI - Organization of sister origins and replisomes during multifork DNA replication in Escherichia coli. AB - The replication period of Escherichia coli cells grown in rich medium lasts longer than one generation. Initiation thus occurs in the 'mother-' or 'grandmother generation'. Sister origins in such cells were found to be colocalized for an entire generation or more, whereas sister origins in slow growing cells were colocalized for about 0.1-0.2 generations. The role of origin inactivation (sequestration) by the SeqA protein in origin colocalization was studied by comparing sequestration-deficient mutants with wild-type cells. Cells with mutant, non-sequesterable origins showed wild-type colocalization of sister origins. In contrast, cells unable to sequester new origins due to loss of SeqA, showed aberrant localization of origins indicating a lack of organization of new origins. In these cells, aberrant replisome organization was also found. These results suggest that correct organization of sister origins and sister replisomes is dependent on the binding of SeqA protein to newly formed DNA at the replication forks, but independent of origin sequestration. In agreement, in vitro experiments indicate that SeqA is capable of pairing newly replicated DNA molecules. PMID- 17914459 TI - Histone chaperones regulate histone exchange during transcription. AB - Transcription by RNA polymerase II is accompanied by dynamic changes in chromatin, including the eviction/deposition of nucleosomes or the covalent modification of histone subunits. This study examined the role of the histone H3/H4 chaperones, Asf1 and HIR, in histone mobility during transcription, with particular focus on the histone exchange pathway, using a dual histone expression system. The results showed that the exchange of H3/H4 normally occurs during transcription by the histone chaperones. Both Asf1 and HIR are important for histone deposition but have a different effect on histone exchange. While Asf1 mediated incorporation of external H3/H4 and renewal of pre-existing histones, HIR opposed it. The balance of two opposing activities might be an important mechanism for determining current chromatin states. PMID- 17914461 TI - HCN4 provides a 'depolarization reserve' and is not required for heart rate acceleration in mice. AB - Cardiac pacemaking involves a variety of ion channels, but their relative importance is controversial and remains to be determined. Hyperpolarization activated, cyclic nucleotide-gated (HCN) channels, which underlie the I(f) current of sinoatrial cells, are thought to be key players in cardiac automaticity. In addition, the increase in heart rate following beta-adrenergic stimulation has been attributed to the cAMP-mediated enhancement of HCN channel activity. We have now studied mice in which the predominant sinoatrial HCN channel isoform HCN4 was deleted in a temporally controlled manner. Here, we show that deletion of HCN4 in adult mice eliminates most of sinoatrial I(f) and results in a cardiac arrhythmia characterized by recurrent sinus pauses. However, the mutants show no impairment in heart rate acceleration during sympathetic stimulation. Our results reveal that unexpectedly the channel does not play a role for the increase of the heart rate; however, HCN4 is necessary for maintaining a stable cardiac rhythm, especially during the transition from stimulated to basal cardiac states. PMID- 17914460 TI - TDP1 facilitates chromosomal single-strand break repair in neurons and is neuroprotective in vivo. AB - Defective Tyrosyl-DNA phosphodiesterase 1 (TDP1) can cause spinocerebellar ataxia with axonal neuropathy (SCAN1), a neurodegenerative syndrome associated with marked cerebellar atrophy and peripheral neuropathy. Although SCAN1 lymphoblastoid cells show pronounced defects in the repair of chromosomal single strand breaks (SSBs), it is unknown if this DNA repair activity is important for neurons or for preventing neurodegeneration. Therefore, we generated Tdp1-/- mice to assess the role of Tdp1 in the nervous system. Using both in vitro and in vivo assays, we found that cerebellar neurons or primary astrocytes derived from Tdp1 /- mice display an inability to rapidly repair DNA SSBs associated with Top1-DNA complexes or oxidative damage. Moreover, loss of Tdp1 resulted in age-dependent and progressive cerebellar atrophy. Tdp1-/- mice treated with topotecan, a drug that increases levels of Top1-DNA complexes, also demonstrated significant loss of intestinal and hematopoietic progenitor cells. These data indicate that TDP1 is required for neural homeostasis, and reveal a widespread requisite for TDP1 function in response to acutely elevated levels of Top1-associated DNA strand breaks. PMID- 17914462 TI - Commensal bacteria modulate cullin-dependent signaling via generation of reactive oxygen species. AB - The resident prokaryotic microflora of the mammalian intestine influences diverse homeostatic functions of the gut, including regulation of cellular growth and immune responses; however, it is unknown how commensal prokaryotic organisms mechanistically influence eukaryotic signaling networks. We have shown that bacterial coculture with intestinal epithelial cells modulates ubiquitin-mediated degradation of important signaling intermediates, including beta-catenin and the NF-kappaB inhibitor IkappaB-alpha. Ubiquitination of these proteins as well as others is catalyzed by the SCF(betaTrCP) ubiquitin ligase, which itself requires regulated modification of the cullin-1 subunit by the ubiquitin-like protein NEDD8. Here we show that epithelia contacted by enteric commensal bacteria in vitro and in vivo rapidly generate reactive oxygen species (ROS). Bacterially induced ROS causes oxidative inactivation of the catalytic cysteine residue of Ubc12, the NEDD8-conjugating enzyme, resulting in complete but transient loss of cullin-1 neddylation and consequent effects on NF-kappaB and beta-catenin signaling. Our results demonstrate that commensal bacteria directly modulate a critical control point of the ubiquitin-proteasome system, and suggest how enteric commensal bacterial flora influences the regulatory pathways of the mammalian intestinal epithelia. PMID- 17914464 TI - Non-covalent interactions in biomacromolecules. AB - Non-covalent interactions play an important role in chemistry, physics and especially in biodisciplines. They determine the structure of biomacromolecules such as DNA and proteins and are responsible for the molecular recognition process. Theoretical evaluation of interaction energies is difficult; however, perturbation as well as variation (supermolecular) methods are briefly described. Accurate interaction energies can be obtained by complete basis set limit calculations providing a large portion of correlation energy is covered (e.g. by performing CCSD(T) calculations). The role of H-bonding and stacking interactions in the stabilisation of DNA, oligopeptides and proteins is described, and the importance of London dispersion energy is shown. PMID- 17914465 TI - Planar carbon radical's assembly and stabilization, a way to design spin-based molecular materials. AB - In this work, we report the first computational study on the assembly and stabilization of a novel kind of radical, i.e., the planar tetracoordinate carbon radical CAl(4)(-). Based on the 6-31+G(d)-UB3LYP, UMP2 and UCCSD(T) calculations on charged [D(CAl(4))M](q-), saturated [D(CAl(4))M(n)] and extended (CpM)(p)(CAl(4))(q) sandwich-like compounds (D = CAl(4)(-), Cp(-); M = Li, Na, K, Be, Mg, Ca), we find that for the six metals, the planar radical CAl(4)(-) can only be assembled in the "hetero-decked sandwich" scheme (e.g. [CpM(CAl(4))](q-)) rather than the traditional "homo-decked sandwich" scheme. Moreover, the low and high spin states of the designed sandwich-like species are perfectly degenerate during assembly. This can be ascribed to the good spin conservation of the CAl(4)(-) deck and the good spatial separation between two CAl(4)(-) decks. Our results show for the first time that the planar radical CAl(4)(-) can act as a new type of spin-embedded "superatom" for cluster assembly when it is assisted by a rigid partner like Cp(-). The good spin-conservation of CAl(4)(-) is very promising for the future design of novel paramagnetic and diamagnetic materials. The ionic, clustering and radical interactions between the two decks are analyzed in detail, which is quite crucial to improve the insight and understanding of the nature and origin of the interactions of the "deck-core-deck" in the metallocenes. Such information is also important in understanding the radical reactions and designing novel spin-based molecular materials. The present study should be expected to enrich the flat carbon chemistry, radical chemistry, metallocene chemistry and combinatorial chemistry. PMID- 17914463 TI - Clustering and synaptic targeting of PICK1 requires direct interaction between the PDZ domain and lipid membranes. AB - Protein interacting with c kinase 1 (PICK1) regulates the trafficking of receptors and ion-channels such as AMPA receptors. Traditionally, the PICK1 PDZ domain is regarded as an adaptor capable of binding to receptors trafficked by PICK1, and the lipid-binding BAR domain functions to tether PICK1 directly to membranes. Here, we show that the PICK1 PDZ domain can directly interact with lipid membranes. The PDZ domain and lipid membrane interaction is mediated by both a polybasic amino-acid cluster and a conserved 'Cys-Pro-Cys' motif located away from the peptide ligand-binding groove. Disruption of the PDZ and lipid membrane interaction totally abolished synaptic targeting of PICK1. Although mutation of the CPC motif did not affect the interaction between PICK1 and AMPA receptors, the mutant PICK1 was unable to cluster the GluR2 subunit of the receptor. In neurons, PICK1 containing the same mutation displayed dramatically compromised capacity in the trafficking of AMPA receptors. Taken together, our findings not only uncovered the novel lipid membrane-binding property of the PICK1 PDZ domain, but also provided direct evidence supporting the functional relevance of the PDZ-lipid interaction. PMID- 17914466 TI - Statistical analysis of intensity fluctuations in single molecule SERS spectra. AB - We have investigated the fluctuations of the intensity and the line intermittency in the surface enhanced Raman spectra of a single iron-protoporphyrin IX molecule. A statistical analysis has revealed a high correlation between the intensity of each frequency couple in the spectrum. Removal of the continuum background has led to a suppression of the correlation at those frequencies where no Raman lines are present. Conversely, we have observed the persistence of a strong correlation at the intensities corresponding to the vibrational modes of the molecule. Further evidence of correlation between the intensities and the background signal indicates that the background is involved in the enhancement mechanism. Moreover, analysis of the Raman line intermittency reveals a random activation of the different molecular vibrational modes. These results can be generally put into relationship to the presence of two different contributions to the intensity fluctuations: one, strictly related to the continuum background, and affecting the whole spectrum, and another one which selectively acts on the various vibrational modes of the molecule. PMID- 17914467 TI - EPR and ENDOR analysis of Fe3+ impurity centers in fluoroelpasolite lattices. AB - Fe(3+) ions in hexagonal and cubic fluoroelpasolite crystals (A(1)(2)B(I)M(III)F(6)) have been investigated in a combined Electron Paramagnetic Resonance (EPR) and Electron Nuclear Double Resonance (ENDOR) study. A detailed analysis of the ENDOR spectra for the nearest (19)F and (23)Na shells in X (9.5 GHz) and Q band (34 GHz) allowed the complex EPR spectra to be disentangled and to determine the spin Hamiltonian parameters for the various S = 5/2 Fe(3+) centres. W-band (95 GHz) EPR measurements as a function of temperature were performed to provide unambiguous evidence about the absolute signs of the Zero Field Splitting (ZFS) and SuperHyperFine (SHF) parameters for Fe(3+) in Cs(2)NaAlF(6) as already determined from the ENDOR work. It could be concluded that all principal (19)F hyperfine values were positive, in agreement with earlier assignments in the literature for related systems. A comparative analysis of the (19)F SHF data for Fe(3+) at a perfectly octahedral site in the cubic crystal, and at two slightly trigonally distorted environments in the hexagonal crystals, indicates that the metal-to-ligand distance changes upon doping. The obtained set of parameters concerning one defect in various analogous environments can furthermore be used to test different methods of theoretical calculations for ZFS and SHF values. PMID- 17914468 TI - Characterization of neutral fragments issued from the photodissociation of protonated tryptophane. AB - New information on the photo-fragmentation of biomolecules is obtained from the detection of neutral and ionic fragments using a time and position resolved coincidence technique that reveals whether an ionic photofragment is associated with one or more neutral fragments. In the case of a sequential dissociation, both fragmentation channels are identified as well as their time ordering. PMID- 17914469 TI - Plasma electrochemistry: electroreduction in a flame. AB - The manipulation of electron transfer reactions at surfaces forms the cornerstone of electrodeposition and processing of materials on substrates with precise control of stoichiometry and oxidation state. However, the utility of this technique, which is mainly carried out in liquid electrolytes, is ultimately limited by the electrolysis of the solvent which limits a potential window to at best 4.8 V in nonaqueous solutions (A. J. Bard and L. R. Faulkner, Electrochemical Methods: Fundamentals and Applications, Wiley, New York, NY, 2nd edn, 2001; ref. 1) and can be up to 6 V in ionic liquids (A. P. Abbott, K. J. McKenzie, Phys. Chem. Chem. Phys., 2006, 8, 4265-4279; ref. 2). A long-sought after goal has been to develop a corresponding technique at the solid/gas interface in the absence of a solvent which will allow in principle a potential window in excess of 100 V (J. M. Goodings, J. Guo, A. N. Hayhurst and S. G. Taylor, Int. J. Mass Spectrom., 2001, 206, 137-151; ref. 3). This extended potential window will enable chemistry at the solid/gas interface that is not possible at the solid/liquid interface. Here we describe a new approach to gas phase electrochemistry using a flame plasma as the electrolyte medium. We demonstrate the controlled electrochemical reduction of Cu(+) to Cu(0) at an electrode surface in a flame environment with resulting deposition of either Cu(2)O or Cu species on conducting diamond electrodes. This approach is novel in that it involves the application of an electrochemical potential difference to change the redox state of surface confined species, not the measurement of flame bore ions (as in flame ionisation detectors). This new technique will permit deposition of films and particles on surfaces with control over the oxidation state of the species. This will provide a valuable enhancement to the capabilities of materials preparation methods such as flame spray deposition. PMID- 17914470 TI - Intracule functional models: I. Angle-corrected correlation kernels. AB - We explore the merits of applying a simple angle-dependent correction to the correlation kernel within the framework of Hartree-Fock-Wigner theory. Based on numerical results for the first eighteen atoms, we conclude that such a correction offers a significant improvement over the action kernel that we and others have explored previously. PMID- 17914471 TI - Probing the evaporation of ternary ethanol-methanol-water droplets by cavity enhanced Raman scattering. AB - Cavity enhanced Raman scattering is used to characterise the evolving composition of ternary aerosol droplets containing methanol, ethanol and water during evaporation into a dry nitrogen atmosphere. Measurements made using non-linear stimulated Raman scattering from these ternary alcohol-water droplets allow the in situ determination of the concentration of the two alcohol components with high accuracy. The overlapping spontaneous Raman bands of the two alcohol components, arising from C-H stretching vibrational modes, are spectrally resolved in stimulated Raman scattering measurements. We also demonstrate that the evaporation measurements are consistent with a quasi-steady state evaporation model, which can be used to interpret the evaporation dynamics occurring at a range of pressures at a particular evaporation time. PMID- 17914472 TI - Small angle X-ray scattering measurements probe water nanodroplet evolution under highly non-equilibrium conditions. AB - Our in situ small angle X-ray scattering (SAXS) measurements yield an unprecedented and detailed view of rapidly evolving H(2)O nanodroplets formed in supersonic nozzles. The SAXS experiments produce spectra in a few seconds that are comparable to small angle neutron scattering (SANS) spectra requiring several hours of integration time and the use of deuterated compounds. These measurements now make it possible to quantitatively determine the maximum nucleation and growth rates of small droplets formed under conditions that are far from equilibrium. Particle growth is directly followed from about 10 micros to 100 micros after particle formation with growth rates of approximately 0.2 to 0.02 nm micros(-1). The peak H(2)O nucleation rates lie between 10(17) and 10(18) cm(-3) s(-1). PMID- 17914473 TI - Chemorheological analysis and model-free kinetics of acid catalysed furfuryl alcohol polymerization. AB - The complete curing of furfuryl alcohol (FA), was studied by chemorheological analysis and model-free kinetics under isothermal and non-isothermal modes. Polymerization of FA under acidic catalysis involves complex reactions, with several steps (such as condensations and Diels-Alder cycloadditions). To account for the polymerization complexity, kinetic analysis of DSC data was performed with a model-free isoconversional method. The obtained E(alpha)-dependencies were closely-correlated with the variation of complex viscosity during curing. Linear condensations are predominant during the early curing stage and are followed by two distinct stages of branching cycloadditions. Gelation and vitrification, identified by rheometric measurements, were associated with a decrease of the overall reaction rate that becomes controlled by diffusion of small oligomers. Before vitrification, the rate of crosslinking is limited by the mobility of longer polymer chains and diffusion encounters a large energy barrier due to the cooperative nature of the motions, leading to higher E(alpha) values. PMID- 17914474 TI - Constraints at the transition state of the D + H2 reaction: quantum bottlenecks vs. stereodynamics. AB - This article presents a quasiclassical trajectory method for the calculation of cumulative reaction probabilities by sampling of the helicity quantum number of the reagents (k). The method is applied to the D + H(2) reaction at various total angular momentum (J) values, and the helicity-resolved quasiclassical cumulative reaction probabilities are compared to their quantum mechanical counterparts. The agreement between the two sets of results is fairly good. In particular, k dependent, J-independent reaction thresholds found with quantum methods are reproduced by the quasiclassical calculations. The shift of these thresholds with increasing k, which has been previously attributed to the quantum bottleneck states taking part in the reaction, is revisited and discussed also in terms of the reaction stereodynamics. PMID- 17914475 TI - Transient behavior of an electrolytic diode. AB - The transient behavior of an electrolytic diode system was studied. A gel-like electrolytic diode was incorporated in a capillary microfluidic chip. The microfluidic platform guaranteed a constant composition of solutions on the diode boundaries. The current responses of the electrolytic diode to step-like changes of the imposed DC electric voltage were measured. Some of these transients were accompanied by a short-time overshoot of electric current density. In order to explain this phenomenon, a mathematical model of the electrolytic diode system was developed. Dynamical analysis of the model equations confirmed the existence of the electric current overshoots. Because the results of the experimental and the numerical transient studies were quite similar, we have explained the physical meaning of three selected overshoots by means of an analysis of the reaction-transport processes inside the electrolytic diode system. The transient experiments carried out in this study indicate that our physical concept of the electrolytic diode system presented in previous papers is correct. PMID- 17914476 TI - Silica-supported chromium oxide: colloids as building blocks. AB - In this contribution, the formation and immobilisation of chromium(iii) hydroxyoxide colloids is investigated. Nano-sized Cr(iii) colloids are generated in situ upon reduction of Cr(vi), dosed to a stirred reactor. The growth of the elementary colloids by the consumption of solved Cr is kinetically unfavorable compared to their aggregation to larger secondary particles, the size of which depends on the concentration of the building block colloids. This aggregation process can be steered by simple process parameters such as dosing rate and concentration of the Cr(vi). The Cr(iii) colloids are immobilised in situ on a support material via precipitation chromatography. Upon drying, the initially amorphose hydroxyoxides are gradually transformed into crystalline Cr(2)O(3) nanoparticles, mainly located at the external surface of the support. This approach opens new opportunities for the synthesis of supported metal oxide catalysts. PMID- 17914477 TI - Time-resolved methods in biophysics. 6. Time-resolved Laue crystallography as a tool to investigate photo-activated protein dynamics. AB - When polychromatic X-rays are shined onto crystalline material, they generate a Laue diffraction pattern. At third generation synchrotron radiation sources, a single X-ray pulse of approximately 100 ps duration is enough to produce interpretable Laue data from biomolecular crystals. Thus, by initiating biological turnover in a crystalline protein, structural changes along the reaction pathway may be filmed by ultra-fast Laue diffraction. Using laser-light as a trigger, transient species in photosensitive macromolecules can be captured at near atomic resolution with sub-nanosecond time-resolution. Such pump-probe Laue experiments have now reached an outstanding level of sophistication and have found a domain of excellence in the investigation of light-sensitive proteins undergoing cyclic photo-reactions and producing stiff crystals. The main theoretical concepts of Laue diffraction and the challenges associated with time resolved experiments on biological crystals are recalled. The recent advances in the design of experiments are presented in terms of instrumental choices, data collection strategy and data processing, and some of the inherent difficulties of the method are highlighted. The discussion is based on the example of myoglobin, a protein that has traversed the whole history of pump-probe Laue diffraction, and for which a massive amount of data have provided considerable insight into the understanding of protein dynamics. PMID- 17914478 TI - Photopolymerization of and patterned fluorescence imaging with a bispyrenyl group containing diacetylene. AB - Photopolymerization of a diacetylene monomer having terminal pyrene groups afforded formation of polydiacetylene nanoparticles in aqueous solvent along with fluorescence quenching of pyrene moieties. PMID- 17914479 TI - Photophysics of 3,5-diphenoxy substituted BODIPY dyes in solution. AB - We have prepared two fluorescent dyes derived from 8-(4-tolyl)-4,4-difluoro-4 bora-3a,4a-diaza-s-indacene with phenoxy and (o-bromo)phenoxy substituents at the 3,5-positions by a novel nucleophilic substitution reaction of the corresponding 3,5-dichloroBODIPY analogue. UV-vis absorption, steady-state and time-resolved fluorimetry have been used to investigate their solvent-dependent photophysical properties. The two BODIPY derivatives show narrow absorption and emission bands and display small Stokes shifts. The substituents at the 3,5-positions (phenoxy in 1 and o-bromophenoxy in 2) have a minor effect on the fluorescence quantum yields (0.16-0.40 for 1, 0.17-0.44 for 2) and lifetimes (1.09-2.51 ns for 1, 1.11 2.78 ns for 2). For both compounds, the fluorescence rate constant equals (1.5 +/ 0.1) x 10(8) s(-1). PMID- 17914480 TI - Photodynamic therapy for cutaneous leishmaniasis: the effectiveness of topical phenothiaziniums in parasite eradication and Th1 immune response stimulation. AB - Photodynamic therapy (PDT) is emerging as a therapeutic modality in the clinical management of cutaneous leishmaniasis (CL). The efficacy of PDT against CL has been demonstrated previously with aminolevulinic acid, although the prolonged terms of therapy were less than ideal, and the search for new photosensitizers (PS) is ongoing. However, phenothiaziniums have demonstrated high parasiticidal effects in vitro. The subject of our investigation is the in vivo activity of two PS, 5-ethylamino-9-diethylaminobenzo[a]phenoselenazinium chloride (EtNBSe) and (3,7-Bis(N,N-dibutylamino) phenothiazinium bromide (PPA904). The results of our comparative analysis of the efficacy of these two phenothiazinium analogues demonstrated a high antiparasitic activity of EtNBSe in vitro, and the higher efficacy of PPA904 in a mouse model of CL. The kinetics of photodestruction are different in parasite and mammalian cells, and with both dyes, the macrophages are more susceptible to photodynamic effects than L. major parasites. As the number of parasites in the lesions undergoes a biphasic change, temporarily increasing on days 2-4 and decreasing on days 5-7, more than one treatment is required within an interval of 5 to 7 days. We have also shown that PPA904-PDT can provide an immunomodulating, dose-dependent efflux on IL-12p70 production. This mechanism could be responsible for promoting a more rapid healing in PPA904 PDT treated mice. Our initial data indicate that phenothiaziniums exhibit a high parasiticidal effect in vivo against CL; this finding may be of use in establishing curative PDT regimens for future clinical trials. PMID- 17914481 TI - Approximating the product spectrum and product concentrations in continuous photochemical reactions. AB - Several approximations to a common photochemical rate law, in which the rate is proportional to the fraction of light absorbed by the reactant chromophore, have been developed to permit the product spectrum to be determined from a sequence of spectra during irradiation that exhibit isosbestic points. The methods were tested on the photolysis of [Cr(NH(3))(6)](3+) and [Cr(en)(3)](3+) (en = ethylenediamine) in water, and [Fe(Et(2)dtc)(3)], tris(diethyldithiocarbamato)iron(III), in CHCl(3). PMID- 17914482 TI - Ship-borne measurements of erythemal UV irradiance and ozone content in various climate zones. AB - Ship-borne measurements of spectral as well as biologically effective UV irradiance have been performed on the German research vessel Polarstern during the Atlantic transect from Bremerhaven, Germany (53.5 degrees N, 8.5 degrees E), to Cape Town, South Africa (33.6 degrees S, 18.3 degrees E), from 13 October to 17 November 2005. Such measurements are required to study UV effects on marine organisms. They are also necessary to validate satellite-derived surface UV irradiance. Cloud free radiative transfer calculations support the investigation of this latitudinal dependence. Input parameters, such as total ozone column and aerosol optical depth have been measured on board as well. Using these measured parameters, the modelled cloudless noontime UVA irradiance (320-400 nm) shows the expected dependence on varying minimum solar zenith angles (SZA) at different latitudes. The modelled cloudless noontime UVB irradiance (290-320 nm) does not show this clear dependence on SZA due to the strong influence of ozone absorption in this spectral range. The maximum daily dose of erythemal irradiance of 5420 J m(-1) was observed on 14 November 2005, when the ship was in the tropical Atlantic south of the equator. The expected UV maximum should have been observed with the sun in the zenith during local noon (11 November). Stratiform clouds reduced the dose to 3835 J m(-1). In comparison, the daily erythemal doses in the mid-latitudinal Bay of Biscay only reached values between 410 and 980 J m(-1) depending on cloud conditions. The deviation in daily erythemal dose derived from different instruments is around 5%. The feasibility to perform ship-borne measurements of spectral UV irradiance is demonstrated. PMID- 17914483 TI - Photophysics, photochemistry, and reactivity: molecular aspects of perylenequinone reactions. AB - Density functional theory (DFT) and time-dependent density functional theory (TD DFT) were used to elucidate the photochemistry and photophysics of eight different perylenequinones (PQ). The objective of this work has been to quantitatively investigate the photodynamic therapeutic potential of this family of compounds and give an overview of their photoreactivity. The effects of solvation were evaluated through single-point calculations using the integral equation formalism of the polarised continuum model. It is concluded that the eight studied perylenequinones can generate singlet oxygen (in aqueous solution) and superoxide radical anions, and that the autoionisation of two nearby PQ molecules is possible. PMID- 17914484 TI - Supramolecular photochemical synthesis of an unsymmetrical cyclobutane. AB - 2-styrylbenzothiazole (1) and cinnamic acid (2) derivatives containing 15-crown-5 ether moieties form a supramolecular assembly in the presence of Ba(2+) cations in acetonitrile. The assembly is stabilized by hydrogen bonding between the heterocyclic N atom of 1 and the proton of the carboxylic group of 2, by sandwich Ba(2+) complex formation between the crown ether moieties of 1 and 2, and by pi pi stacking interactions. Irradiation of solutions containing these supramolecular complexes leads to highly specific formation of an unsymmetrical cycloadduct. This investigation provides an interesting example of supramolecular control of [2 + 2]-photocyclization in solution. PMID- 17914485 TI - Measuring the lifetime of singlet oxygen in a single cell: addressing the issue of cell viability. AB - Singlet molecular oxygen, O(2)(a(1)Delta(g)), has been detected from single neurons and HeLa cells in time-resolved optical experiments by its 1270 nm phosphorescence (a(1)Delta(g)--> X(3)Sigma(-)(g)) upon irradiation of a photosensitizer incorporated into the cell. The cells were maintained in a buffered medium and their viability was assessed by live/dead assays. To facilitate the detection of singlet oxygen, intracellular H(2)O was replaced with D(2)O by an osmotic de- and rehydration process. The effect of this insult on the cells was likewise assessed. The data indicate that, in the complicated transition from a "live" to "dead" cell, the majority of our cells have the metabolic activity and morphology characteristic of a live cell. Quenching experiments demonstrate that the singlet oxygen lifetime in our cells is principally determined by interactions with intracellular water and not by interactions with other cell constituents. The data indicate that in a viable, metabolically-functioning, and H(2)O-containing cell, the lifetime of singlet oxygen is approximately 3 micros. This is consistent with our previous reports, and confirms that the singlet oxygen lifetime in a cell is much longer than hitherto believed. This implies that, in a cell, singlet oxygen is best characterized as a selective rather than reactive intermediate. This is important when considering roles played by singlet oxygen as a signaling agent and as a component in events that result in cell death. The data reported herein also demonstrate that spatially-resolved optical probes can be used to monitor selected events in the light-induced, singlet-oxygen-mediated death of a single cell. PMID- 17914486 TI - Real-time fluorescence monitoring of phenothiazinium photosensitizers and their anti-mycobacterial photodynamic activity against Mycobacterium bovis BCG in in vitro and in vivo models of localized infection. AB - An objective was to explore the photodynamic activity of two cationic photosensitizers (PS) (benzo[a]phenothiazinium chloride and benzo[a]phenoselenazinium chloride) against Mycobacterium bovis BCG both in vitro and in a murine model of BCG-granuloma. The hypothesis being tested in this study was that cationic molecules could best interact with the negatively charged membrane of BCG as a model for mycobacterial infection. Cells in culture were incubated with various concentrations of PS and subsequently illuminated using a 635 nm diode laser. Dark- and light-induced killing profiles were generated as a function of fluence and dye concentration. In vivo, local injection of the PS into subcutaneous Mycobacterium-induced granuloma sites in murine model was followed by red light illumination of the same area. A special microscope was fabricated for real-time in vivo fluorescent microscopy to monitor EtNBS delivery to subcutaneous murine granulomata. Both PS demonstrated good in vitro antimycobacterial photodynamic activity with varying degrees of toxicity under dark conditions. Real time in vivo monitoring of benzophenothiazine chloride in the mouse model indicated that this fluorescent photosensitizer was delivered rapidly to the subcutaneous granuloma site. In vivo, photosensitizer specific dark- and photo-toxicities depended on the structure, concentration of the photosensitizer and the light dose utilized. Cationic phenothiazine photosensitizers are promising candidates for use in anti-mycobacterial PDT for localized diseases such as cutaneous and pulmonary granulomata. PMID- 17914487 TI - Chronic obstructive pulmonary disease in a new concept. AB - Chronic Obstructive Pulmonary Disease (COPD) is a preventable and treatable disease with some significant extrapulmonary effects that may contribute to the severity in individual patients. Its pulmonary component is characterized by airflow limitation that is not fully reversible. The airflow limitation is usually progressive and associated with an abnormal inflammatory response of the lung to noxious particles or gases. Although COPD is a nonspecific term referring to a set of conditions that develops progressively because of a number of different disease processes, it most commonly refers to patients with chronic bronchitis and emphysema and to a subset of patients with asthma. Several different definitions were proposed for COPD in time. COPD is not asthma but can coexist with asthma, the other major airways obstructive disease caused by airway inflammation. Inflammation underlying in asthma has characteristic features, distinct of that from COPD. Longitudinal studies revealed the heterogeneous character of COPD. The pathological hallmarks of chronic obstructive pulmonary disease (COPD) are inflammation of the small airways (bronchiolitis) and destruction of lung parenchyma (emphysema). International guidelines stress the importance of accurately discriminating between asthma and chronic obstructive pulmonary disease. Although characteristic pathological features have been described for both conditions, their discriminatory power has never been systematically assessed. This might be rectified by improving pathological definitions. PMID- 17914488 TI - The immunohistochemical profile of the adenocarcinoma of upper gastric pole. AB - Although gastric adenocarcinoma continue to be the second continues to be the second cause of death worldwide, its incidence and mortality appear to have decreased in recent decades. Despite this decline, adenocarcinomas from proximal stomach tend to be more frequent during the last three decade. Adenocarcinomas with this location it seems that are a different, specific subtype of gastric carcinoma. The purpose of this study was to clarify the differences between gastric adenocarcinomas from upper and distal gastric pole using the immunohistochemistry. For this reason, we investigate histopathological and immunohistochemically 77 cases of upper gastric pole adenocarcinoma selected from a number of 472 gastric tumors. The immunohistochemistry was performing only in 32 cases by ABC technique with the following primary antibodies: Cytokeratin 7, Cytokeratin 19, Epithelial Membrane Antigen (EMA), Carcinoembryonic Antigen (CEA), Lysozyme, Vimentin, p53 protein, CD34 and Ki67 antigen. The acquired results do not distinguish a peculiar immunohistochemically profile unlike distal gastric adenocarcinomas. Nevertheless, we pointed out the predominance of diffuse adenocarcinomas type according to Laurens classification, which immunohistochemically were strong positive to cytokeratins, EMA, CEA and lysozyme. Moreover, investigation of some antigens likes lysozyme, p53, Ki67 and CD34 seems to be useful for prognostic estimation of carcinoma with this topography. PMID- 17914489 TI - Simultaneous demonstration of mast cells and blood vessels by the combined method CD34--alcian blue-safranin in lip tumors. AB - The aim of the study was to evaluate the mast cell-blood vessel relationship using double staining CD34/AAS. Sections from 14 cases with lip tumors have been stained with Hematoxylin-Eosin. On additional sections from each case, we highlighted blood vessels by immunohistochemistry for CD34 antigen using the method LSAB2-HRP/DAB, followed by alcian blue-safranin stain for mast cells. We quantified the density, distribution and the mast cell types as well as the correlation with the number of blood vessels. All cases have been positive for both staining. We observed a significant correlation between the number of vessels and the mast cells (p = 0.003). In one case, we observed the mast cells stained with safranin (red), the vascular density being less than the mast cells density. Our results confirmed the data from the literature with respect to the large number of mast cells observed in the malignant tumors. The increased vascular density together with the mast cell density suggests a correlation between these two elements in the tumor angiogenesis, possibly though the VEGF secretion. The CD34/AAS stain is a quick and simple method and it allows an optimal correlation between the number of mast cells and blood vessels on the same section. The type of mast cells correlated with microvessel density is a powerful argument towards the involvement of the mast cells in the tumor angiogenesis of the malignances of the lips. PMID- 17914490 TI - Genetic and clinical considerations in six cases with neurofibromatosis type 1. AB - Neurofibromatosis type 1 (NF1) is an autosomal dominant disorder, caused by mutations in the NF1 gene. The NF1 gene encoding neurofibromin protein, which is strongly expressed in the nervous system and with the role as a negative regular of the ras proteins signal. All six cases with neurofibromatosis type 1 were clinical and laboratory investigated. The frequently symptoms are "cafe au lait" spots and neurofibromas. In two cases, the disease is associated with essential hypertension and, in other two cases with kyphoscoliosis. The novo mutations in NF1 gene cause the disease in three cases, and in other three cases, the mutation is inherited (two cases on father side and one case on mother side). PMID- 17914491 TI - Bone abnormalities occurring in the follow-up of the patients with neurofibromatosis type 1. AB - Neurofibromatosis type 1 (NF1), also called von Recklinghausen disease or peripheral neurofibromatosis, is a common autosomal dominant disorder characterized by multiple neurofibromas, "cafe au lait" spots and Lisch nodules of the iris with a variable clinical expression. Osseous anomalies appeared in the patients with NF1 including dysplasia, scoliosis and pseudoarthrosis. We propose a research of the osseous involvement at 11 patients, seven female and four male with ages from 9 to 60 at which the cutaneous aspect has the complete form, hyperpigmented spots and cutaneous neurofibromas and only more than six "cafe au lait" spots. All the patients suffered radiological exams, CT- and MRI scan. The results were different from case to case from the extreme severe deformations, especially at the children, to clinical unapparent osseous involvement, incidental found or with occasion of our investigation. CONCLUSIONS. The patients with NF1 has osseous abnormalities specific of the disease, like dysplasia, scoliosis, pseudoarthrosis, often gentle but sometimes extremely severe. The most severe osseous involvement are presented in the cases when these development early in the childhood. Other times the osseous abnormalities are clinical asymptomatic, their finding been clinical incidental. We want to have a separate mention for the maxillary and mandible involvement, which according to our information is not a rare form. PMID- 17914492 TI - Scanning electron microscopy of vascular corrosion casts--standard method for studying microvessels. AB - Scanning electron microscopy (SEM) of vascular corrosion casts (microvascular corrosion casting/SEM method) is a standard method, which allows three dimensional visualization with good resolution of the normal and abnormal microvessels, including the capillaries of various organs and tissues. SEM of vascular corrosion casts can obtain qualitative as well as quantitative informations important to anatomists, pathologists and clinicians. Considering these, the history, the advantages and the main steps of this technique including general morphological characteristics of vascular casts observed in SEM are reviewed in this paper. Corrosion casts done by the author representing the microvascular organization of the rat liver and kidney observed in SEM are, also, presented. PMID- 17914493 TI - Arcuate foramen of atlas: incidence, phylogenetic and clinical significance. AB - Arcuate foramen is less known trait of the human atlas vertebra formed by a delicate bony spiculum, which arches backward from the posterior end of the superior articular process. Examination of 1044 human atlas vertebra revealed that the trait was present in 13.8% of the samples. The mean length of the arcuate foramen form was 7.16 mm on the left side and 9.99 mm on the right side in bilateral positive samples while it was 8.14 mm and 9.26 mm respectively in unilateral positive samples. The mean vertical height of this foramen was 6.57 mm on the left side and 6.52 mm on the right side in bilateral positive samples while it was 4.91 mm and 5.38 mm respectively in unilateral positive samples. The sides did not show any statistical significant differences. The importance of the arcuate foramen lies in the external pressure it may cause on the vertebral artery as it passes from the foramen transversarium of the first cervical vertebra to the foramen magnum of the skull. PMID- 17914494 TI - Angiogenesis and progesterone receptor status in primary breast cancer patients: an analysis of 158 needle core biopsies. AB - Formation of new blood vessels from a preexisting vascular bed (angiogenesis) is a complex multistep process, which may also permit metastasis. Progesterone receptor is a surrogate marker for ER activity and has been used as an additional predictive factor for hormonal therapy in breast cancer. To investigate how tumor angiogenesis correlates with progesterone receptor (PR) status in breast carcinoma diagnosed on core biopsy, microvessels were counted (and graded the density of microvessels) within the initial invasive carcinomas of 158 patients. Using light microscopy, the number of microvessels was counted manually in a subjectively selected hot spot (in the most active areas of neovascularization per 400x field), and their values were separated as above or below median (low and high), without knowledge of the outcome in the patient or any other pertinent variable. When the mean values of MVD of the various groups defined by PR status were compared, significant difference was noted (P = 0.008557). When tumors were classified as high or low MVD, based on a cut-off value (30.70175 microvessels/mm(2)), cases with high MVD were significantly more numerous. MVD did show a relationship with groups defined by PR status (P = 2.03076E-05). The correlation of angiogenesis with PR status may be a potential therapeutic target for the treatment and prevention of breast cancer, using antiangiogenic molecules. PMID- 17914495 TI - Ultrastructural analysis of the endometrial mucosa of female patients at menopause with hormonal substitutive therapy (TSH). AB - The objective of the paper was to observe the ultrastructural aspects of the endometrial biopsies taken from female patients at post-menopause with substitutive hormonal therapy (TSH). Material and methods. A number of three endometrial biopsies were taken from female patients at post-menopause with TSH. The ultrastructural analysis was carried out with the help of the electronic microscope Philips ME 301 using classical electronic microscopy methods. Results and discussions. The ultrastructural analysis has highlighted the presence of cuboidal and columnar epithelial cells, with basally situated nuclei, well represented cellular organelles, some cells having at the apical pole microvilli. At the electronic microscope, three types of epithelial cells are described, at the level of the endometrial mucosa of the woman who is in a fertile period: secretory cells (cells with an average electronic density with microvilli on the luminal surface), ciliated cells and clear cells (cells with a low electronic density). These cells have certain ultrastructural characteristics and of receptivity towards the steroid hormones. The stroma is axial with elongated cells with oval nuclei, with nucleoli and with smooth or undulated membrane. Conclusions. The ultrastructural aspects suggest the presence at endometrial level of epithelial active glandular cells, secretory cells and stromal active cells at female patients at post-menopause with TSH. PMID- 17914496 TI - Macro- and microanatomic factors involved in the flat bones morphogenesis from the craniofacial system. AB - The authors present a study both upon eight crania of dead born fetuses having the vertex-coccis distance between 25-29 cm and a human embryocephalic extremity of 18 cm. The crania were examined in the lateral, vertical, frontal, occipital, basal norms. Sagittal sections were performed upon the embryo, and then those sections were microscopically examined after HE staining. We concluded that the cranial arch bones morphogenesis and the facial complex is a long-time development process, which initially started during the early embryogenesis, and it is completed as an adult. Determining factors of the flat bones osteogenesis are the following: vascular, muscle, extracell (mesenchyma) and neuronal factors (rhombencephalon presence). PMID- 17914497 TI - The study of CD20 and CD45.Ro antibodies in the inflammatory infiltrate involved in acne and seborrheic dermatitis. AB - The aim of our study was to evaluate the role of CD20 and CD45.Ro antibodies in acne and seborrheic dermatitis. A number of 20 patients with papular, pustular or nodular acne and another 20 patients with seborrheic dermatitis were available for our study. We removed bioptic material from all of them and we perform histochemical and immunohistochemical processing within the Laboratory of Histology, Histopathology and Immunohistochemistry of the University of Medicine and Pharmacy of Craiova. In acne, we could reveal a positive CD45.Ro immunomarking in rare lymphoid cells situated in the middle derma at a distance from the affected pilosebaceous follicle and in the inflammatory infiltrate subepidermically, and also a negative immunomarking in the inflammatory cells from the proximity of the affected pilosebaceous follicle. In patients with seborrheic dermatitis we noticed a positive immunomarking infiltrate of the papillary derma and a positive immunomarking of membrane for CD45.Ro in many lymphoid cells of the inflammatory infiltrate situated in the papillary derma predominantly disposed perivasculary. Conclusions. The absence of the cells marked with CD45.Ro in the proximity of the pilosebaceous follicle interested in acne excludes the direct participation of B- and T-lymphocytes in the perifollicular inflammatory process, though the T-lymphocytes can be revealed in a small number at a distance from the affected follicle. The inflammatory infiltrate from the seborrheic dermatitis proved to be rich in positive CD45.Ro cells and poorer in positive CD20 cells. PMID- 17914498 TI - Expression of cytokeratin MNF116 and vimentin in pleural serous effusions. AB - The purpose of this study is to evaluate the value of cytokeratin (CK) MNF116 and vimentin in the differential diagnosis of malignant pleural effusions. There were evaluated smears from 30 patients with pleural effusions stained with May Grunwald Giemsa and Papanicolaou techniques for the routine cytological diagnosis. Additional smears were immunostained with CK MNF116 and vimentin using LSAB2 technique. Two independent observers evaluated all smears. Smears were classified first by cytological examination in seven cases (23.33%) as benign, and in 23 cases (76.67%) as malignant pleural effusions. Mesothelial cells expressed CK MNF116 in 96.67% (29/30) of cases and vimentin in 33.33% (10/30) of cases. Malignant cells expressed CK MNF116 in 52.17% (12/23) of cases and vimentin in 30.43% (7/23) of cases. The pattern of immunostaining was diffuse cytoplasmic. In conclusion, CK MNF116 and vimentin may be used as a part of the panel of antibodies for differential diagnosis of malignant pleural effusions with primary unknown. PMID- 17914499 TI - Duplicated omohyoid muscle and its clinical significance. AB - The omohyoid muscle is one of the infrahyoid muscles with superior and inferior bellies. Variations of this muscle are clinically important because of its relation to the internal jugular vein as well as its significance in radical neck dissection. A duplicated omohyoid was observed during routine cadaver dissection, which is attached along with the usual inferior belly of the omohyoid into the transverse scapular ligament. Knowledge of anomalies of this muscle is important to minimize the complications during the surgical procedures of cervical region. PMID- 17914500 TI - Anorectal melanoma. Case report and review of the literature. AB - We report a case of primary anorectal melanoma, a very rare malignant tumor in this localization. The patient, a 57-year-old female, complained for one year of symptoms, which were attributed to hemorrhoids. An abdomino-perineal resection was performed. Pathologic examination revealed an anorectal melanoma with numerous lymph node metastases and an unusual metastatic deposit in a uterine leiomyoma. The pathogenesis, pathology, principles of treatment and prognosis of this entity are briefly discussed. PMID- 17914501 TI - Giant retroperitoneal sarcomas. AB - Retroperitoneal sarcomas are rare malignant tumors, which are developing from mesenchymal stem cells residing in muscle, fat, and connective tissues. Underlying the rarity of this kind of tumors in general population, the aim of this paper is to present three cases of retroperitoneal sarcomas operated in Surgical Department of Military Hospital of Craiova (a retroperitoneal liposarcoma, a dedifferentiated liposarcoma and a malignant fibrous histiocytoma). From clinical point of view, we note the poverty of symptoms and non-specificity of these and a great tolerability of retroperitoneal space that offers the possibility for a great development of the tumor. CT-scan and MRI are the best investigations for diagnosis but surgical exploration is the best way for a good evaluation of these tumors. From histological point of view, we try to present new features about these kinds of tumors in order to classify them. Results of surgery correlated with complementary therapies were good without per operative mortality or postoperative morbidity but we noted a recidive of one tumor (with different histological pattern) 21 months after the surgical intervention. The rarity of retroperitoneal sarcomas, combined with the vast array of histologic subtypes, has complicated our understanding of these tumors and impeded the development of effective therapies. PMID- 17914502 TI - Small cell carcinoma of the urinary bladder--a new case report. AB - Primary pure small cell carcinoma of the urinary bladder is an extremely rare and highly aggressive tumor with an average five-year survival rate of less than 10% as cited by multiple case reports. It accounts for about 0.5-1% of all bladder tumors. We present the case of a 44-years-old man, smoker (10 cigarettes/day) hospitalized in the Department of Urology, from the "Prof. dr. Th. Burghele" Hospital, Bucharest, for one month intermittent hematuria. Ultrasonography showed a sessile tumoral mass, sized 37/30mm. Transurethral resection of the tumor mass was performed and tissue fragments were sent to the pathologic lab to establish the histologic type, the degree of differentiation and invasion. Fragments of the tumor were fixed in 10% formaldehyde, paraffin embedded and processed as standard technique; the sections were stained with HE, VG and immunohistochemically with: CROMO, EMA, NSE, CD56, NK1, p53 and betaHCG. The microscopic examination reveled a tumor proliferation composed of two distinct components: extensive small cells areas and foci of typical low grade (G2) papillary urothelial carcinoma. The small cell are uniformly, round, with increased nucleo-cytoplasmic ratio, eosinophyl cytoplasm, hyperchromatic nuclei, finely granular chromatin and inconspicuous nucleoli. Immunohistochemical stains showed diffuse positive staining of the small cell component for CROMO, EMA, NSE, CD56, NK1 and urothelial carcinoma component stained focally for betaHCG. The rate of cell proliferation was increased (p53 - 80% positive reaction). Conclusions. A diagnosis of small cell carcinoma coexisting with low-grade urothelial carcinoma was established. Because of aggressive behavior and distinct treatment, the pathologist should watch out for the presence of small cell carcinoma component. PMID- 17914503 TI - The etiology of Mycobacterium avium subspecies paratuberculosis in Crohn's disease. AB - The etiology of Crohn's disease (CD) has drawn heated controversy in the literature. Compelling evidence in the literature has accumulated lately that could incriminate the Mycobacterium avium subspecies paratuberculosis (MAP), the well-known agent of John's disease in cattle. The evidence is isolation of the organism or its DNA and RNA, detection of the anti-MAP antibodies in Crohn's patients, increasing incidents of CD in areas close to the cattle pastures, and the possibility of treating the disease with the antibiotics. The group that favors the immune dysregulation theory considered this evidence circumstantial due to the variations in these reports. The treatment of CD with humanized anti tumor necrosis factor-alpha antibodies is considered great endorsement to the immune dysregulation theory. The endless debate could jeopardize public health rather than bring a final solution. Reconciliation between the 2 theories appears inevitable in view of possible classification of this disease as a zoonotic. PMID- 17914504 TI - Determination of sensitivity of male Wistar rats to an equal dose of ketamine/xylazine injection at anesthetic dose in a chronic model of hypernatremia in comparison with control group. AB - OBJECTIVE: To determine the sensitivity to an equal dose of ketamine/xylazine injection at anesthetic dose in a chronic model of hypernatremia. METHODS: This study was conducted at the Department of Physiology, Zanjan University of Medical Science, Zanjan, Iran in 2004. Sixty male Wistar rats, weighing 250 +/- 20 g were randomly allocated to 3 groups. The control group was provided with tap water, and first and second test groups consumed 1% and 2% salt concentrations for 144 hours. One hundred mg/kg ketamine and 10 mg/kg xylazine were used as an anesthetic agent. The measured anesthetic parameters comprises of righting reflex latency, required time for establishment of animal's immobility, immobility period, required time for appearance of animal's mobility and complete re establishment of the righting reflex. RESULTS: The required time for inhibition of the righting reflex and animal's mobility in the second group was significantly shorter than the first and control groups. Immobility period, required time for appearance of animal's mobility and complete re-establishment of the righting reflex in the second group were significantly longer than the first and control groups. CONCLUSION: Hypernatremia increases the speed of transition from different steps of ketamine/xylazine anesthesia with significant delay in immobility period and recovery from anesthesia in rats, hence, anesthetic dose reduction in hypernatremia is necessary. PMID- 17914506 TI - Analysis of HumFABP2 as a polymorphic human genetic marker in the Turkish population. AB - OBJECTIVE: To examine the allele frequencies of HumFABP2 locus in 155 individuals from different regions of Turkey. METHODS: The study was carried out in Cumhuriyet University Hospital, Sivas, Turkey, between March and June 2006. The allele and genotype frequencies for HumFABP2 were determined by polymerase chain reaction (PCR) using the manufacturer's recommended protocol, and using the commercially available Macherey-Nagel DNA isolation kit. The PCR amplification was carried out in a Perkin-Elmer GeneAmp PCR System 9600 thermal cycler following the manufacturer's recommendations. The allele frequencies in the Turkish population was computed, and the heterozygote rate was calculated. RESULTS: In this population study of 155 samples, we found 75 (48.39%) heterozygote and 80 (51.61%) homozygote. The results showed heterozygotic cases as 150/250 bp, and homozygotic cases as 150 bp. CONCLUSION: Allele frequency data of HumFABP2 as a PCR-based genetic marker could be used in identity testing to estimate the frequency of a multiple PCR based profile in the Turkish population. PMID- 17914505 TI - The protective effect of ethyl pyruvate on lung injury after burn in rats. AB - OBJECTIVE: To investigate the effects of administered ethyl pyruvate (EP), a novel anti-inflammatory agent, on oxidoinflammatory and apoptotic pathways in the lung tissue of rats in a full-thickness burn model. METHODS: The study took place in Ankara Research and Training Hospital Animal Laboratory, Turkey in June 2006. Thirty-two rats were randomly divided into 4 groups in equal numbers as sham, burn, sham+EP, and burn+EP. The burn model, used produced a full thickness burn of the 30-35% of the total body surface area. Ethyl pyruvate was administered as 40 mg/kg intraperitoneally. Rats were sacrificed after 24 hours, acute lung injury (ALI) was evaluated by direct light microscopy and apoptosis was evaluated by caspase-3 staining. Oxidoinflammatory events were evaluated by determining the tissue levels of myeloperoxidase (MPO), lipid peroxidation products, and nitrite. RESULTS: No significant difference was observed in lung tissue nitrite and malondialdehyde levels among the study groups. Histopathological results revealed that ALI and apoptosis were significantly higher in the burn group and EP prevented this effect. Similar results were obtained in tissue MPO levels. CONCLUSION: Ethyl pyruvate is a novel, potent anti-inflammatory agent. This agent prevented leukocyte infiltration, ALI, and apoptotic loss of the lung tissue in thermal injury. PMID- 17914507 TI - Renin-angiotensin system polymorphisms and renal graft function in renal transplant recipients. AB - OBJECTIVE: To analyze the role of 3 polymorphisms of the renin-angiotensin system (RAS) in renal transplant recipients (RTRs) and correlate them with graft function. METHODS: The present study was performed in the Drug Applied Research Center, Tabriz Medical University, Tabriz, Iran from September 2003 to December 2005 on 108 RTRs (66 males and 42 females, with a mean age of 37.34 +/- 4.97 years) with stable allograft function (creatinine < or =2.2 mg/dl). Following the DNA extraction from the blood leukocytes, the genotypes of the angiotensin converting enzyme (ACE I/D), angiotensinogen (ANG M235T), and angiotensin II type 1 receptor (ATR1 A1166C) were determined by polymerase chain reaction. The magnitude of clearance of creatinine (ClCr) in the setting of each of the above RAS polymorphisms was determined. The ClCr was measured by modification of diet in renal disease formula. Values were expressed as mean +/- SD; p< or =0.05 was considered to indicate statistical significance. RESULTS: There was no association of each genotype of the RAS alone with ClCr, serum urea, cyclosporine through level and the degree of urinary protein excretion rate. However, patients with the DD genotype of angiotensin converting enzyme + CC genotype of angiotensin II type I receptor polymorphisms had lower ClCr (p=0.05) and a higher urinary protein excretion rate (p=0.03). Other combination genotypes of RAS had no effect on allograft function. Interestingly, the percent of hypertensive patients in the C allele (70%) was more than the A allele (30%) of ATR1 polymorphism (p=0.04). CONCLUSION: Although none of the single gene polymorphisms of the RAS affected renal allograft function, combinations of these genotypes were associated with the outcome of allograft function. PMID- 17914508 TI - Lupus nephritis. Clinicopathological correlation. AB - OBJECTIVE: To classify all renal biopsies of lupus nephritis patients presenting in the last 10 years, according to the modified World Health Organization (WHO) classification using the facilities of light, fluorescent, and electron microscopy. To assess the activity and chronicity indices of renal biopsies according to the National Institute of Health protocol, and to correlate the histological findings with the clinical features of the patients presented up to the time of biopsy. METHODS: Seventy-eight patients with lupus nephritis, biopsied over 10 years between January 1995 to December 2005 in King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia were reviewed by 2 histopathologists with the assistance of a nephrologist. RESULTS: The predominant histological type was WHO class IV. Patients of this class were more commonly associated with microhematuria, elevated proteinuria, and renal insufficiency. Active and chronic lesions were more likely to occur in patients of class III/IV. These patients were also more likely to have evidence of clinical renal disease than patients in class II. There was a significant association between nephrotic syndrome and class V lupus nephritis. CONCLUSION: Although the clinical and pathological correlation demonstrated a significant relationship between underlying histopathology and the clinical course of the patient, however, the biopsy findings did not uniformly correlate with the clinical features. Moreover, the status prediction of lupus nephritis patients based on clinical information alone was significantly enhanced by information obtained from renal biopsy. PMID- 17914509 TI - Clinicopathological study of primary gastric lymphoma. AB - OBJECTIVE: To present a histopathologic and immunohistochemical analysis of primary gastric lymphomas that was reclassified according to the new World Health Organization classification of lymphoid neoplasms. METHODS: We reviewed the morphological and immunohistochemical features of 28 patients with gastric lymphomas, diagnosed in the Department of Pathology at the University Hospital of Tishreen University, Lattakia, Syria, during the period 1994-2003. Specimens were obtained from endoscopic and surgical biopsies. The immunohistochemical study was performed to analyze the immunophenotype of these lymphomas. RESULTS: Patients were aged 17-71 years. There was a slight predominance of females (male to female ratio, 13:15). Seventeen of the patients had tumors mainly located in the gastric antrum. Histologically, the most common lymphoma was of mucosa-associated lymphoid tissue (MALT) type (20 patients), also with diffuse large B-cell lymphoma (7 patients), and anaplastic large cell lymphoma (one patient). CONCLUSION: Our study demonstrates the different patterns of gastric lymphomas in Lattakia, Syria during a 10-year period in 28 Syrian patients, and reveals that the most primary gastric lymphomas are B-cell MALT lymphomas. PMID- 17914510 TI - Determination of human papillomavirus type 16 genotype and construction of cloning vector pTZ57R encoding HPV16 E7 gene. AB - OBJECTIVE: To isolate and construct a cloning vector containing the human papillomavirus (HPV)16-E7 gene as a target for application as a DNA vaccine. METHODS: The study was performed in 2005 in Iran. The E7 gene, one of the most important HPV oncoproteins and a target molecule for therapeutic vaccines, was amplified by polymerase chain reaction (PCR). The PCR product was cloned into a suitable cloning vector and confirmed by colony-PCR, restriction enzyme analysis, and sequenced. RESULTS: The desired plasmid was sequenced and indicated 99% homology with those mentioned in the Genbank. CONCLUSION: The Iranian HPV16 E7 gene sequence is very similar to other sequences in the Genbank, and it can be used as a candidate gene in a therapeutic vaccine for Iranian patients with cervical cancer. PMID- 17914511 TI - Hepatitis B and C virus infections in patients with hemophilia in Zahedan, southeast Iran. AB - OBJECTIVE: To determine the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections in patients with hemophilia in Zahedan, Iran. METHODS: From March 2003 to January 2006, we evaluated 81 hemophiliac patients in Zahedan Hemophilia Center, southeast Iran, for hepatitis C virus antibody (HCV-Ab) and hepatitis B surface antigen (HBsAg), and evaluated the prevalence of HBV/HCV co infection. RESULTS: The seroprevalence of HCV was 29.6%, and the HBsAg was positive in 4.9%. Four cases had HCV and HBV co-infection. All of the infected patients were unknowingly treated with contaminated plasma products before the middle of 1996. CONCLUSION: All hemophiliacs, especially patients who have been treated with unheated clotting factor concentrates, should be evaluated for HCV and HBV infections. PMID- 17914512 TI - Variations in Serratia marcescens differentiation using different primers. AB - OBJECTIVE: To study primer sequences (1060, 1247, 1254, 1281, 1283, and 1290) for random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). METHODS: Twenty-four clinical Serratia marcescens (S. marcescens) isolates were obtained over a 6-month period from April 2002 to September 2002 from hospitals in the Fars province of Iran. Six primers were used due to S. marcescens genome properties, and RAPD-PCR was carried out. The results were subjected to unweighted pair-group method analysis using NTSYSpc 2.02. The primers were blasted with the S. marcescens genome, and the primers efficiency was estimated. RESULTS: The results of blast primers with S. marcescens genome sequence showed that primer 1283 had the highest homology and primer 1290 had the lowest homology. Comparing the resulted dendrograms showed that the pattern of the primers to separate isolates was closely related to their sequence homology with the genome and their amount of guanine and cytosine nucleotide content. CONCLUSION: There are clear differences in RAPD-PCR results when different primers are used, and it is recommended to consider genomic properties of an organism to design a primer for RAPD-PCR. PMID- 17914513 TI - Monopolar electrodissection versus cold dissection tonsillectomy among children. AB - OBJECTIVE: To compare cold dissection to monopolar tonsillectomy in terms of operative time, intraoperative bleeding, post operative bleeding, and pain. METHODS: The study included 100 children who underwent tonsillectomy between January 2002 and January 2004. This study was carried out at King Abdul-Aziz University Hospital, Riyadh, Kingdom of Saudi Arabia. Patients were randomly selected to have either the right or left tonsils removed by either technique (cold dissection or monopolar dissection technique). We compared both techniques in each side on the same patient. RESULTS: There was no significant difference in operative time between both sides by either technique, however, when the patients were grouped into < or =3 minutes and >3 minute groups, the result showed that a greater number of patients were operated up on < or =3 minutes by diathermy procedure compared to dissection method (44 versus 67) (p=0.0011). Blood loss was minimal with the diathermy technique; averaging 25.37 ml compared to 88.5 ml for cold dissection tonsillectomy. There was a significant increase in postoperative pain on the first postoperative day in the diathermy group compared to the cold dissection group (27% versus 12%) (p=0.0151). However, there was no significant difference in pain between both sides in the first 24 hours and from the 2nd to 10th postoperative days. CONCLUSION: Monopolar dissection tonsillectomy is a safe technique. It significantly reduces the operative time and intraoperative blood loss. However, it causes more pain on the 1st post operative day, while on the rest of the days until the 10th post operative day, there was no significant difference in pain between two sides. PMID- 17914514 TI - Standard surgical versus percutaneous dilatational tracheostomy in intensive care patients. AB - OBJECTIVE: The present prospective randomized trial compared surgical tracheostomy (ST) and percutaneous dilatational tracheostomy (PDT) in intensive care unit (ICU) patients in terms of outcomes and complications. METHODS: Between January 2003 and December 2005 tracheostomies were performed on critically ill ICU patients in Medical Faculty Hospital in Prague, with a random allocation of 105 patients for ST and 100 for PDT. RESULTS: The 2 groups did not differ significantly in terms of basic demographic characteristics or length of endotracheal intubation prior to the procedure. Following the procedures, the 2 groups did not differ significantly in terms of the time required for decannulation, decannulated patients or mortalities. Post-mortem examination showed that both groups were similar in terms of placement of the tracheostomy tube. Surgical tracheostomy was found to take longer time to perform than PDT (p<0.001). In terms of early postoperative complications, PDT was associated with a higher rate of postoperative bleeding compared to ST (p=0.0302). CONCLUSION: Percutaneous dilatational tracheostomy is a simpler and faster technique to perform, but is associated with a higher occurrence of early complications, particularly postoperative bleeding. PMID- 17914515 TI - Suturing of the nasal septum after septoplasty, is it an effective alternative to nasal packing? AB - OBJECTIVE: To discuss and compare the results of suturing the nasal septum after septoplasty with the results of nasal packing. METHODS: A prospective study, which was performed at Prince Hashem Military Hospital in Zarqa, Jordan and Prince Rashed Military Hospital in Irbid, Jordan between September 2005 and August 2006 included 169 consecutive patients that underwent septoplasty. The patients were randomly divided into 2 groups. After completion of surgery, the nasal septum was sutured in the first group while nasal packing was performed in the second group. RESULTS: Thirteen patients (15.3%) in the first group and 11 patients (13%) in the second group had minor oozing in the first 24 hours, 4 patients (4.8%) had bleeding after removal of the pack in the second group. Four patients (4.8%) developed septal hematoma in the second group. Two patients (2.4%) had septal perforation in the second group. One patient (1.1%) in the first group, and 5 patients (5.9%) in the second group had postoperative adhesions. Five patients (5.9%) were found to have remnant deviated nasal septum in each group. The operating time was 4 minutes longer in the first group. CONCLUSION: Septal suturing after septoplasty offers the following advantages: elimination of discomfort for the patients, minimal complications, the outcome is almost the same as with nasal packing, and finally the hospital stay is less than with nasal packing. Therefore, suturing of the nasal septum after septoplasty should be a preferred alternative to nasal packing. PMID- 17914516 TI - Effect of use of mitomycin C on the outcome of Choanal atresia repair. AB - OBJECTIVE: To investigate the effectiveness of topical mitomycin C (MMC) on outcome of endoscopic repair of choanal atresia (CA) in cases that had never had this operation and when nasal stent was not used. METHODS: Endoscopic repair of CA was performed in 20 children at King Abdul-Aziz University Hospital, Riyadh, Kingdom of Saudi Arabia between January 1999 and October 2005. Intra-operative application of topical MMC (0.4 mg/mL for 4 minutes) was carried out in 13 children, and 7 children did not receive MMC. The follow-up period ranged between 9 months and 6 years. The association between the 2 categorical variables was investigated using Fisher's exact test. RESULTS: Sixty-nine percent of the cases receiving MMC had a successful outcome compared with 57% of non-MMC cases, however, the difference was not statistically significant (p=0.230). In unilateral CA, 70% of those who received MMC had a successful outcome compared with 60% of those who did not; again the difference was not statistically significant (p=0.566). Successful outcome was considered when the posterior choana was patent during the successive follow up, without the need for revision surgeries. CONCLUSION: We did not demonstrate a statistically significant difference between the percentages of children treated with MMC versus no-MMC who remained patent after surgery. PMID- 17914517 TI - Arthroscopy versus arthrocentesis. A retrospective study of disc displacement management without reduction. AB - OBJECTIVE: To compare the outcome of arthroscopic surgery and arthrocentesis of internal derangement (disc displacement without reduction) of the temporomandibular joint (TMJ). METHODS: The charts of 48 patients with TMJ derangement and treated with arthroscopy or arthrocentesis were included in this study carried out in a specialized private clinic in Beirut, Lebanon, between January 2001 and July 2005. Inclusion criteria included patients with a history of clicking followed by a sudden onset of limited mouth opening without clicking, a complaint of TMJ pain with mouth opening or chewing difficulty, or both, or a positive magnetic resonance imaging diagnosis of TMJ disc displacement without reduction. Patients with limited mouth opening caused by only muscle spasm, prior TMJ surgery, bilateral joint involvement, or serious systemic diseases were excluded from the study. Twenty-eight patients underwent arthroscopy (group one) and 20 patients were treated with arthrocentesis (group 2). RESULTS: Both methods showed a significant reduction in pain and an increase in maximal mouth opening on follow up (p<0.01). There was no statistical difference between the methods. CONCLUSION: Further research should be conducted before one can definitely determine if real benefits are achieved through surgery in TMJ articular disorders. Within the limits of this study, less invasive procedures are highly recommended. PMID- 17914518 TI - Measurement of coronary sinus blood flow after first anterior myocardial infarction with transthoracic echocardiography and its correlation with wall motion scoring index. AB - OBJECTIVE: To measure the coronary sinus blood flow (CSBF) and coronary sinus velocity time integral (CSVTI) via transthoracic echocardiography (TTE) in patients with acute myocardial infarction (AMI) in association with the left ventricular ejection fraction (LVEF), and wall motion scoring index (WMSI). METHODS: In this case-control study, 20 patients with anterior AMI and 20 healthy individuals as the control group, were studied in 6 months period from March to September 2005 in Madani Heart Center, Tabriz, Iran. All patients received the same treatment for AMI (such as fibrinolytic). The CSBF, CSVTI, WMSI, and tissue Doppler imaging (TDI) data were obtained via TTE and compared between the 2 groups. RESULTS: Baseline variables were similar between the 2 groups (p>0.05). The CSBF in AMI group was 287.8 +/- 128 ml/min and in the control group was 415 +/- 127 ml/min (p=0.001). Also, CSVTI was significantly lower in AMI group than control group (11.16 +/- 2.85 and 17.56 +/- 2.72 mm, p=0.003). There was a significant correlation between CSBF and LVEF (r=0.52, p=0.01), WMSI (r=-0.77, p=0.0001) and CSBF and in-hospital mortality (r=0.58 p=0.03), also between CSVTI and LVEF (r=0.85, p=0.0001), WMSI (r=-0.57, p=0.0009) and in-hospital mortality rate (r=0.69, p=0.02). The CSBF and CSVTI had a good correlation with TDI findings: peak early diastolic velocity in the myocardium and peak systolic velocity in the myocardium). CONCLUSION: Our study demonstrated a good correlation between measured CSBF and CSVTI by 2D- Doppler TTE and LVEF, WMSI, in hospital mortality and TDI findings. PMID- 17914519 TI - Adding remifentanil to propofol and etomidate in cardioversion anesthesia. AB - OBJECTIVE: To compare their effects on cardiorespiratoy and recovery parameters and side effects. METHODS: This study was performed in The Ministry of Health Ankara Numune Research and Training Hospital, Ankara, Turkey, from January to May 2005. The 40 American Society of Anesthesiology II/III patients were randomized into 2 groups. All patients received remifentanil 0.75 microgram.kg(-1); and then received either etomidate 0.1 mg.kg(-1) (group E, n=20) or propofol 0.5 mg.kg(-1) (group P, n=20). Cardiorespiratory data, induction time, recovery parameters, pain scores, number of shocks (NS), total amount of energy used (TE), side effects, and patient/cardiologist satisfaction were recorded. RESULTS: Induction time and recovery parameters were shorter in group P. No differences were seen between the groups in NS and mean TE required. In group P, a statistically significant decrease in mean blood pressure occurred after induction and returned to its baseline levels in 6 minutes. After cardioversion over 2 minutes, the respiratory rates were decreased significantly more in group P when compared with group E. Two patients in group P became apneic and needed assisted ventilation. Pain scores, side effects and patient/cardiologist satisfaction were similar in both groups. No patients in either group had myoclonus. CONCLUSION: We can induce hypnosis with propofol 0.5 mg.kg(-1) or etomidate 0.1 mg.kg(-1) by adding remifentanil 0.75 microgram.kg-1 in cardioversion anesthesia. Although recovery parameters were longer in group E, and cardiorespiratory parameters were less stable in group P, their usage with remifentanil was both acceptable for cardioversion anesthesia. PMID- 17914520 TI - Growth charts for Saudi children and adolescents. AB - OBJECTIVE: To establish reference growth charts for Saudi children and adolescents. METHODS: Multi-stage probability sampling of a cross-section of Saudi children and adolescents residing in all 13 regions of the Kingdom. Family interviews, and physical examinations of children and adolescents from birth to 19 years of age were conducted over a 2-year period (2004-2005). Only healthy children and adolescents were included for the measurement of length/stature, weight, and head circumference. All measurements were performed by trained physicians and nurses according to World Health Organization guidelines. Percentile construction and smoothing were performed using the LMS (lambda, mu, sigma) methodology. RESULTS: Determination of the standard measures for normal physical growth in a sample representing healthy Saudi children and adolescents from birth to 19 years of age. CONCLUSION: The results of this study present the most comprehensive and up-to-date reference growth charts for Saudi children and adolescents. Therefore, the authors recommend the use of these charts to replace older charts or those belonging to other countries. PMID- 17914521 TI - Prevalence and trends in obesity among school boys in Central Saudi Arabia between 1988 and 2005. AB - OBJECTIVE: To determine the trends in body fatness and obesity among Saudi primary schoolboys using the data from 2 cross-sectional studies conducted in 1988 and 2005. METHODS: Two sets of data were analyzed. The first set (n=1082) was conducted in 1988 and the second (n=702) set was conducted in 2005. Both studies used multistage random samples involving primary-school boys aged 6-14 years from Riyadh, Kingdom of Saudi Arabia. Measurements included weight (Wt), height (Ht), biacromial (BA) and bi-iliac (BI) widths, triceps (T), subscapular (S) skinfolds, S/T ratio, body mass index (BMI), body fat percentage (fat%), lean body mass (LBM), and the proportion of obese boys (fat% > or =25% of Wt). RESULTS: From 1988-2005 there were significant increases in all variables except LBM. The lowest changes were observed in body structures (Ht, BA, and BI) and the highest were in body fatness (T, S, and fat%). During this 17-year period, the mean BMI standard deviation increased from 16.5 +/- 2.1 to 18.0 +/- 4.0 kg/m2 and fat percentage increased from 13.2 +/- 4.7 to 19.7 +/- 10.0%. In addition, S/T ratio increased by 13.5%, indicating shifts toward central obesity over time. However, the biggest increase was seen in the proportion of obese schoolboys (from 3.4% in 1988 to 24.5% in 2005). CONCLUSION: Findings indicate rising trends in BMI, body fatness, central obesity, and prevalence of obesity among Saudi schoolboys over the last 2 decades. Increased obesity prevalence among Saudi children is a major public health concern. PMID- 17914522 TI - Malnutrition, and anthropometric and biochemical abnormalities in end-stage renal disease patients. AB - OBJECTIVE: To detect and compare the degree of malnutrition as well as the anthropometric and biochemical abnormalities among male and female patients with end-stage renal disease (ESRD). METHODS: A cross-sectional study was carried out at hemodialysis units in 7 Jordanian hospitals, between 2004 - 2005. Two hundred and seventeen ESRD outpatients who underwent hemodialysis were recruited using the convenience-sampling technique. A subjective global assessment (SGA) was used to assess the degree of malnutrition in both male and female patients. Anthropometric and biochemical measurements were also assessed in all patients. RESULTS: The SGA suggested that 50% of females were well nourished while 75% of the males were moderately to severely malnourished. The measured anthropometric variables showed a significant decrease in both male and female patients, except for triceps skinfold thickness. The hemoglobin as well as blood creatinine level were lower among females. In the biochemical parameters, there was a significant decrease in hemoglobin, albumin, and total protein blood levels in male patients. However, a significant decrease in hemoglobin only was detected in female patients with advanced malnutrition. CONCLUSION: The nutritional status determined by SGA indicated a higher proportion of male patients with moderate to severe malnutrition as compared to female patients. Most of the anthropometric parameters could be used to assess the nutritional status of hemodialysis patients. The noticeable hemoglobin reduction, which increased with malnutrition degree, was significant in both male and female patients, while serum albumin and total protein reduction was significant among male patients only. PMID- 17914523 TI - Seasonal variation in the incidence of congenital hypothyroidism in Isfahan, Iran. AB - OBJECTIVE: To evaluate the seasonal variations in the incidence of congenital hypothyroidism (CH) in the screening program of CH in Isfahan, Iran. METHODS: In this study, we compiled the data obtained retrospectively from CH screening results of 113282 neonates from 17 maternity hospitals in Isfahan, Iran from June 2002 to December 2005. The seasonal variation in the incidence of CH, as well as its monthly incidence was analyzed using all the diagnosed cases of CH. RESULTS: From the 113282 neonates referred for CH screening, 358 neonates were diagnosed with CH, showing an overall incidence of 3.1/1000 live birth. There was no significant difference in the seasonal variation of CH (p=0.5). According to the monthly distribution analysis, the incidence of CH was higher in the second month of summer (Mordad) and lower in the last month of autumn (Azar)(p=0.04). CONCLUSION: The high incidence of CH in Mordad, the second month of summer, supports the hypothesis that certain environmental factors such as intrauterine viral infections (which commonly exhibit seasonal variations in incidence), exposure to chemical compounds, differences in climate and so forth, may play a role in the etiology of this disorder. Awareness of the monthly distribution of CH incidence could help us identify associated environmental factors and aid in the development of preventative strategies. Further, it would for the appropriate allocation of resources within the screening programs. PMID- 17914524 TI - Gigantic enlargement of the thymus gland. AB - True massive thymic hyperplasia is a very rare entity, characterized by an increase in the size and weight of the thymus gland, without an apparent cause. Surgery has been required in patients with severe respiratory distress. We present an idiopathic true massive thymic hyperplasia in a 5-month-old boy. PMID- 17914525 TI - Gut metastasis from breast carcinoma. AB - Breast cancer is the second most common malignancy in women. Common sites of metastases include the liver, lung, bone, and the brain. Metastases to the gastrointestinal tract are rare with patients presenting with small-bowel perforation, intestinal obstruction, and gastrointestinal bleeding. Here we report a case of a Saudi female presenting with invasive lobular carcinoma and ileo-cecal junction metastasis. PMID- 17914526 TI - Chronic granulomatous disease with recurrent hepatic abscesses in an adult. AB - Chronic granulomatous disease CGD is a condition of inability to deal with bacterial and fungal infections, due to defective respiratory burst in neutrophils leading to recurrent cutaneous and visceral infections. Usually a disease of childhood, but patients nowadays survive to adulthood, and diagnosis might be difficult if not considered. We describe a 20-year-old female with previously undiagnosed CGD, presenting with recurrent cutaneous and hepatic abscesses. PMID- 17914527 TI - Congenitally corrected transposition of great arteries with ischemic symptoms in middle age. AB - Congenitally corrected transposition of the great arteries CCTGA is a rare congenital disease first described by Von Rokitansky in 1875. Transposition of the great arteries comprises 2.6 - 7.8% of all cases of congenital heart disease, and if uncorrected, is commonly fatal in the first year of life. Patients with corrected transposition of the great arteries without associated defects may remain undiagnosed until adult life. Symptoms occur rarely before the fourth and fifth decades, when rhythm disturbance, left atrioventricular valve regurgitation, and moderately impaired systemic ventricular function cause congestive cardiac failure. We report here a case of drug overdose with ischemic symptoms, and CCTGA without associated anomalies in a 40-year-old male. PMID- 17914528 TI - Isotretinoin in acne agminata. AB - Acne agminata is an asymptomatic papulopustular eruption. This condition typically occurs in young adults. The eruption generally runs a self-limited course, but disfiguring scars can occur. Histological examination shows scattered dermal granulomas composed of epitheloid and some giant cells with central caseation. A variety of agents such as wide-spectrum antibiotics, oral steroids, dapsone, and clofazimine have been used with varying degrees of success. Herein, we report 2 Caucasian males with acne agminata, successfully treated with isotretinoin. PMID- 17914529 TI - An invasive thymoma complicated with airway stenosis causing sudden hypoxia during surgery. PMID- 17914530 TI - The effect of propolisis and mesalazine on bacterial translocation in an experimental colitis model. PMID- 17914531 TI - Peroneal tendon dislocation. A tendon protective and bone preserving technique of stabilization. PMID- 17914532 TI - A histopathological study of chronic granulomatous lymphadenitis. PMID- 17914533 TI - Endoscopic dacryocystorhinostomy for primary nasolacrimal duct obstruction. PMID- 17914534 TI - Hepatosplenic abscess in brucellosis. PMID- 17914535 TI - Clinical quiz: a family's name for a familial disease. PMID- 17914536 TI - Re: A fatal case of Behcet's disease with rare complications. PMID- 17914537 TI - Re: Management of hypertensive patients in primary health care setting, auditing the practice. PMID- 17914538 TI - Medical journals: old, not senescent. A tribute to Revista Medica de Chile on its 135th anniversary. AB - Medical journals, like the Revista Medica de Chile, are the product of a long history of printing, literary styles, and scientific inquiry. They would not have developed without the craft of paper making, the invention of movable type, the need to communicate ideas and observations in a timely and digestible format, and the growth of "natural philosophy" during the European Renaissance and Enlightenment. The proliferation of scientific and medical periodical literature during Europe's industrialization in the nineteenth and twentieth centuries followed the emergence of medical science and care as a separate discipline and the creation of common and worldwide professional expectations and credentials. Medical journals continue to evolve with the advent of digital paperless publishing. Prognostication remains uncertain but it seems to me unlikely that printed paper, so called "hard copy, "journals will become extinct any time soon. PMID- 17914539 TI - [Coronary angiography: indications, results and complications in 5.000 consecutive patients]. AB - BACKGROUND: There are guidelines about equipment, premises, personnel, indications and complications rates for coronary angiography, that every center performing this procedure should adhere. AIM: To report the experience with 5.000 coronary angiographies and to assess the compliance of the center with the current guidelines. MATERIAL AND METHODS: Prospective registry of 5.000 patients aged 60+/-11 years (3.475 males) subjected to coronary angiography since 1992, assessing all aspects of the procedure with special emphasis on complications. RESULTS: The indications for 80% of procedures was suspected coronary atherosclerosis. The main risk factors were hypertension and smoking. Coronary atherosclerosis was demonstrated in 62%, mainly one vessel disease. These were two deaths due to the procedure (0.04%), three patients (0.06%) had an acute myocardial infarction or a stroke. These figures are lower than referential values. CONCLUSIONS: In this center, coronary angiography is a safe procedure, with complications rates that are even lower than referential values. PMID- 17914540 TI - [Chest pain unit: first experience in Chile]. AB - BACKGROUND: In large series, nearly 60% of admissions for suspected acute coronary syndrome (ACS) had a non-coronary etiology of the pain. However, short term mortality of non recognized ACS patients, mistakenly discharged from the emergency room is at least twice greater than the expected if they would had been admitted. The concept of a chest pain unit (CPU) is a methodological approach developed to address these issues. AIM: To evaluate the efficacy of a CPU in the emergency room of a general hospital for evaluation of acute chest pain. MATERIAL AND METHODS: Prospective study of patients with chest pain admitted in the CPU. After a clinical, electrocardiographic and laboratory evaluation with cardiac injury serum markers, patients were stratified in three risk groups, based on the likelihood of ACS of the American Heart Association. High probability patients were admitted to the Coronary Unit (CU) for treatment. Moderate probability patients remained in the CPU for further evaluation and low probability patients were discharged with telephonic follow-up. RESULTS: Of 407 patients, 35, 30 and 35% were stratified as high, intermediate and low probability ACS, respectively. Among patients admitted with high probability, 73% had a confirmed ACS diagnosis. Among intermediate probability patients, 86% were discharged after an evaluation in the CPU without adverse events in the follow-up. CONCLUSION: Structured risk evaluation approach in a CPU improves the management of acute chest pain, identifying high probability patients for fast admission and start of treatment in a CU and allowing safe discharge of low probability ones. PMID- 17914541 TI - [Anthropometry, body composition and functional limitations in the elderly]. AB - BACKGROUND: Functional limitations limit the independence and jeopardize the quality of life of elderly subjects. AIM: To assess the association between anthropometric measures and body composition with functional limitations in community-living older people. MATERIAL AND METHODS: Cross-sectional survey of 377 people > or = 6 5 years old (238 women), randomly selected from the SABE/Chile project. Complete anthropometric measurements were done. Handgrip muscle strength was measured using dynamometers. Body composition was determined using Dual-Energy X-Ray Absorptiometry. Functional limitations were assessed using self reported and observed activities. RESULTS: Body mass index was strongly associated with fat mass (men r =0.87; women r =0.91) and with lean mass (men r =0.55; women r =0.62). Males had significantly greater lean mass (48.9 kg vs 34.9 kg), and bone mass than females (2.6 kg vs 1.8 kg) and women had higher fat mass than men (26.3 kg vs 22.9 kg). The prevalence of functional limitations was high, affecting more women than men (63.7% vs 37.5%, p <0.01). Functional limitations were associated with lower handgrip strength in both sexes. In the multiple regression models, with functional limitations as dependent variable and anthropometric measures as contributing variables, only hand grip strength had a significant association (negative) with functional limitations in both genders. Age was also a significant risk factor for functional limitations among women. CONCLUSIONS: Hand grip strength was strongly and inversely associated with functional limitations. Handgrip dynamometry is an easy, cheap and low time consuming indicator for the assessment of functional limitations and the evaluation of geriatric interventions aimed to improve functional ability. PMID- 17914542 TI - [Long-term continuous positive airway pressure (CPAP) use in obstructive sleep apnea]. AB - BACKGROUND: The use of CPAP is the conventional therapy for the Obstructive Sleep Apnea Syndrome (OSAS). AIM: To establish the proportion of OSAS patients with CPAP indication who use it in the long-term and to determine predictive factors of adherence. PATIENTS AND METHODS: Structured survey through personal interview or phone call, applied to all patients who had polisomnographically documented OSAS and CPAP titration at the Pontificia Universidad Catolica de Chile Sleep Disorders Center between January 1998 and November 2001. Patients were divided in "Permanent Users" (Group I) and "Non Users" (Group II). RESULTS: Of 440 patients that fulfilled inclusion criteria, 162 (32.8%) answered the survey. Seventy percent (114/162) of these patients had used CPAP and 86% (98/114) continued to use it at the time of the interview, with a mean follow-up of 12.3 months (Group I 60.5% of the interviewed population). Group II included 64 patients (39.5% of the interviewed patients). Among the latter, 25% (16/64) stopped using CPAP due to intolerance, 25% (16/64) could not use it because of economical restrictions, 20.5% (13/64) preferred other therapies and 29.5% (15/64) merely refused treatment. Predictors of long-term use of CPAP were number of respiratory events (p <0.01), hypersomnolence (p <0.01) and age (p =0.03). CONCLUSIONS: Approximately two thirds of patients with OSAS, with an indication for CPAP, continue to use this therapy in the long term in a Chilean population. This figure is similar to other reports from abroad. It is also possible to identify predictive factors for CPAP discontinuation in this population. PMID- 17914543 TI - [An enquiry on client opinions about quality of care in a public hospital]. AB - BACKGROUND: The level of satisfaction of patients with the services provided by public hospitals is becoming an important issue for health authorities. AIM: To analyze an enquiry applied to patients, about the level of satisfaction with the service provided by a public regional hospital in Chile. MATERIAL AND METHODS: An enquiry was applied to 150 ambulatory (76% women) and 50 hospitalized patients (56% women). It included 28 questions about management aspects of health care and 20 questions about client satisfaction. Data was analyzed using factorial analysis and multiple regressions. RESULTS: The manners of hospital personnel were the factor with the highest impact on the perception of patients about global quality of services. Attention timing was the most influential aspect on global satisfaction. CONCLUSIONS: There results confirm those from abroad and can orient an adequate management of public hospital. PMID- 17914544 TI - [Cryoablation of pulmonary veins as complementary treatment of atria I fibrillation in valvular surgery]. AB - BACKGROUND: The Cox MAZE III operation for the treatment of atrial fibrillation (AF) is complex and consumes significant operative time. Cryoablation of the pulmonary veins (CPV) is a simpler alternative for patients that require concomitant valvular surgery. AIM: To evaluate CPV in patients with AF submitted to valvular surgery. PATIENTS AND METHODS: Twenty one patients had simultaneous valvular surgery and CPV, 81 % of them had permanent AF for an average of 5 years. Twenty patients had mitral valve disease. The etiology was rheumatic in 14. Average left atrial diameter was 60 mm. In 7 patients the mitral valve was replaced, in 5 it was repaired, in 7 both mitral and aortic valve were replaced, in 1 the mitral valve was repaired and the aortic valve was replaced and in 1 only the aortic valve was replaced. A combined transeptal and superior approach was used for all patients. The CPV was performed after the valvular procedure with cryothermy at -60 degrees C for 2 minutes with two 15 mm cryoprobes applied simultaneously. RESULTS: CPV increased surgical time by 10 to 20 minutes. Operative mortality was 4.8% (1 patient). One patient developed a pericardial effusion and another a complete heart block that required a permanent pacemaker. All patients improved their functional class. At the end of an average 10.5 months of follow-up, 50% of patients were in normal sinus rhythm and 25% persisted in AF. The remaining patients were in some type of regular rhythm. CONCLUSIONS: CPV as a complementary procedure in patients with AF undergoing valvular surgery had good results to abate AF. It restored normal sinus rhythm in 50% of the cases, with low morbidity and mortality and little increment in surgical time. PMID- 17914545 TI - [Glycemic and insulin indices of tube feeding formulas in healthy adults]. AB - BACKGROUND: In acute illnesses, plasma glucose levels are often increased and generally parallel the severity of stress. Hyperglycemia caused by reduced insulin sensitivity and reduced insulin secretion is associated with increased susceptibility to infections. Maintaining blood glucose levels at or below 110 mg/dl reduces morbidity and mortality in critically ill patients. AIM: To measure the glucose and insulin responses of four commercially available enteral formulas compared with a standard meal reference product. MATERIAL AND METHODS: The glycemic index (GI) and the insulin index (II) were determined in a randomized, cross over protocol in 38 healthy volunteers between 18 and 46 years of age. Each subject underwent five tests: three with the standard meal (bread) and two with the study products. The enteral formulas were Clinutren HPR (whole protein of high protein value), Crucial (casein peptide based formula), Peptamen, (whey peptide based formula), Glytrol (formula for diabetics with whole protein with fiber). Each study product was evaluated 10 times. RESULTS: The diabetic formula and the high protein energy dense formulas induced a significantly lower GI (p <0.02) compared with the standard meal. The GI response did not appear to be due to enhanced insulin secretion. The other tested formulas had lower GI than the standard meal, but in addition they exhibited increased II The whey based peptide formulation produced the highest insulin response (p <0.03). CONCLUSIONS: Both GI and II are related to the concentration, form and type of protein contained in the enteral formula. The whey peptide formulation produced a low GI with the highest insulin index. Based on the low GI of these enteral products, all can be useful to provide nutritional support during metabolic stress, without adding an additional challenge to blood glucose management. PMID- 17914547 TI - [Glutathione metabolism during postnatal development of the rat lung]. AB - BACKGROUND: Significant changes in lung antioxidants occur in preparation for birth. Little is known about physiological regulation of antioxidants in the postnatal period. AIM: To study the glutathione system in the lungs during postnatal development. MATERIAL AND METHODS: In the lungs of 7, 15, 21, 50 and 70 days old Sprague-Dawley rats we measured total and oxidized glutathione content as well as the activity of the limiting enzyme in glutathione synthesis (y-GCS) and of the enzymes glutathione peroxidase (GPx) and glutathione reduciase (GRd). RESULTS: Between 7 and 15 days the activities of GPx and GRd increase 32% and 26%, respectively (p <0.001). Whereas GPx activity remains high throughout the rest of the study period, GRd activity decreases progressively reaching adulthood values at 7 days. y-GCS activity shows a gradual increase that reaches significance at 50 days when it doubles values observed at 7 days (p <0.05). A significant correlation was found between GPx and GRd activities over the entire period (r =0.62, p <0.0001). Strength of the correlation is age dependent due to the differences in time course of the enzyme changes. Whereas total GSH does not change, oxidized glutathione decreases from 7% at 7 and 15 days to 4% later on (p <0.01). CONCLUSIONS: The activity of several enzymes involved in glutathione metabolism increases during postnatal development of the rat lung. Interpretation of lung responses to injurious agents needs to be done taking into consideration the physiological regulation of antioxidants during postnatal development. PMID- 17914546 TI - [Management of hazardous waste in a hospital]. AB - BACKGROUND: An inadequate management of hospital waste, that have toxic, infectious and chemical wastes, is a risk factor for humans and environment. AIM: To identify, quantify and assess the risk associated to the management of hospital residues. MATERIAL AND METHODS: A cross sectional assessment of the generation of hazardous waste from a hospital, between June and August 2005, was performed. The environmental risk associated to the management of non-radioactive hospital waste was assessed and the main problems related to solid waste were identified. RESULTS: The rate of generation of hazardous non-radioactive waste was 1.35 tons per months or 0.7 kg/bed/day. Twenty five percent of hazardous liquid waste were drained directly to the sewage system. The drug preparation unit of the pharmacy had the higher environmental risk associated to the generation of hazardous waste. The internal transport of hazardous waste had a high risk due to the lack of trip planning. The lack of training of personnel dealing with these waste was another risk factor. CONCLUSIONS: Considering that an adequate management of hospital waste should minimize risks for patients, the hospital that was evaluated lacks an integral management system for its waste. PMID- 17914548 TI - [Prevalence of cardiovascular risk factors in adult from Talca, Chile]. AB - BACKGROUND: Cardiovascular disease (CVD) has several traditional risk factors (RF), and some of them are potentially modifiable. AIM: To determine the prevalence of most common risk factors in adult population in Talca in Central Chile. SUBJECTS AND METHODS: We studied 1007 subjects aged 18 to 74 years (66% women), living in Talca, selected by a probability sampling. They answered a questionnaire and anthropometry, blood pressure, lipid profile and blood glucose were measured. RESULTS: Thirty seven percent of subjects smoked, 70.1% had a body mass index over 25 kg/m(2) and 41% had an abnormally high circumference. High blood pressure was found in 37% (36% of these subjects were unaware of this condition), 44.5% had hypercholesterolemia, 21.5% had low HDL cholesterol, 40.1% had hypertriglyceridemia and 26.3% had high blood glucose levels. CONCLUSIONS: There is a high prevalence of risk factors for CVD in adult population from Talca. The prevalence of risk factors is higher than that reported by the 2003 National Health Survery. PMID- 17914549 TI - [Neurological involvement in Wegener's granulomatosis: report of one case]. AB - Wegener's granulomatosis is a systemic necrotizing vasculitis that affects medium size and small vessels. Neurological involvement occurs in 22% to 54% of patients, mainly in the form of mononeuritis multiplex. Central nervous system involvement is reported in only 2% to 8% of the cases. We report a 42-year-old male who presented with headache, diplopia, third and sixth cranial nerve palsies and left eye amaurosis associated to mass located in the Meckel cavum and diffuse meningeal involvement. A biopsy of the mass disclosed a chronic granulomatous necrotizing inflammation with Langhans giant cells. A chest CAT scan showed three cavitated lung nodules and ANCA antibodies were positive in a titer of 1:80. Treatment with steroid and cyclophosphamide was started and cranial nerve palsies resolved and the number and size of lung nodules decreased. The patient was lost from follow up. PMID- 17914551 TI - [Retroperitoneal bronchogenic cyst: Report of one case]. AB - Retroperitoneal cystic tumors are uncommon. More than two thirds are malignant. Benign lesions include lipomas and leiomyomas, among others. Bronchogenic cysts are congenital anomalies that result from an abnormal budding of the primitive foregut, and are most commonly found in the mediastinum. Occasionally they can be seen in the skin, subcutaneous tissue, in the pre-sternal and pericardial areas, but those located under the diaphragm are extremely rare. We report a 24 year-old female who consulted for abdominal pain. An abdominal ultrasound disclosed a retroperitoneal cystic lesion, that was confirmed with a CAT scan. The patient was operated, finding a cyst that was adhered to the pancreas. A distal pancreatectomy and cyst excision were done. The pathological examination of the lesion showed a bronchogenic cyst. The patient was discharged ten days after surgery. PMID- 17914550 TI - [Hematopoietic stem cell transplantation for patients with Wiskott-Aldrich syndrome]. AB - BACKGROUND: Wiskott-Aldrich syndrome (WAS) is an X linked congenital disease that presents as eczema, thrombocytopenia and immune deficiency. The only curative procedure for this illness is hematopoietic stem cell transplant (HSCT), preferably from a healthy HLA identical sibling donor. Cord blood is becoming an excellent alternative as stem cell source from unrelated donors. AIM: To report our experience with HSCT in children with WAS. PATIENTS AND METHODS: Six boys with WAS diagnosed at 1 to 6 months of age were transplanted at our institution. All of them developed eczema and thrombocytopenia. Two had episodes of severe bleeding and three had repetitive infections (two with recurrent pulmonary infections and one a recurrent otitis). Three patients had a positive family history. Two received HSCT from sibling donors and four from unrelated cord blood donors at 7 months to 4 years of age. RESULTS: AH 6 patients had full hematopoietic engraftment after transplantation. Three had mild chronic graft versus- host disease which responded to immune suppressive therapy. One patient died of cytomegalovirus related pneumonia 111 days after grafting. The other 5 patients are alive and healthy 11 to 104 months after transplantation. CONCLUSIONS: HSCT is an effective treatment for patients with WAS. The procedure should be done as soon as diagnosis is confirmed and before life threatening infections occur. PMID- 17914552 TI - [Which are the causes of death among Chileans today?. Long-term perspectives]. AB - During the last decades, Chile experienced substantial socioeconomic, epidemiological and demographic changes. These resulted, among other consequences, in a deceleration of population growth, a notorious decrease in fertility rates, and one of the most rapid and deepest drop in general and infant mortality rates in the Latin American region. These changes resulted in a sustained increase of life expectancy and a substantial ageing of the Chilean population. This process is also changing the disease burden of the population. Infectious and perinatal diseases lost relevance as major causes of mortality, and have been replaced by chronic non transmissible diseases, specifically cardiovascular conditions and cancer, that are becoming the main causes of death. High blood pressure, cardiovascular risk, hypercholesterolemia, diabetes, overweight and obesity, smoking, sedentary lifestyle and depression will have a great impact on health conditions during the XXI century. These factors and a persistent social inequity will hinder the efforts to reduce the impact and consequences of chronic non transmissible, diseases in the Chilean population. PMID- 17914553 TI - [Analysis of electronic references in a volume of Revista Medica de Chile]. AB - BACKGROUND: Publications in electronic format using Web sites facilitates and extends the access to information from any place in the world. Nevertheless, the use of electronic references still lacks the rigor with which the printed reference is handled. AIM: To analyze the citation of electronic references in Revista Medica de Chile. MATERIAL AND METHODS: Twelve issues of volume 132 were analyzed, quantifying the number of electronic references. Their accuracy was assessed considering the elements available in the Web site and access date. RESULTS: Seventy six electronic references were detected. Of these, 28 (37%) incorporated available in, 76 (100%) indicated the Web site and 38 (50%) indicated the access date. CONCLUSIONS: Electronic references are still scanty and incomplete in Revista Medica de Chile. PMID- 17914554 TI - Prognostic value of estrogen receptor and progesterone receptor tumor expression in Danish ovarian cancer patients: from the 'MALOVA' ovarian cancer study. AB - Estrogen and progesterone are important hormones secreted by the ovary acting through specific receptors. Tumor tissue expression profiles of these have demonstrated prognostic value in malignancies such as breast, uterine and prostate cancer. In this study, including tissue samples from 773 Danish patients with an ovarian tumor, we evaluated whether estrogen receptor (ER) and progesterone receptor (PR) expression correlated with clinico-pathological parameters, and a possible prognostic impact on ovarian cancer (OC) patients was investigated. Using tissue array and immunohistochemistry, we analyzed the ER and PR expression levels in tissues from 582 women with OC and 191 women with low malignancy potential (LMP) ovarian tumors. Our results demonstrated that ER was expressed in 30 of the 191 LMP tumors (16%) and in 207 of the 582 OC (36%). PR was expressed in 38 LMP tumors (20%) and in 115 OC (20%). For both tumor types an excess of positive tumors was found in the serous compared to the mucinous subtype (p< or =0.00001). The frequency of ER expression-positive OC increased with increasing FIGO stage (p=0.0003), and the frequency of PR-positive tumors increased with increasing histological grade (p=0.0006). In a Cox survival analysis, a tissue ER and PR expression 10% or higher was found to imply an independent significant advantageous course of patient disease-specific survival (ER: hazard ratio (HR), 0.80; 95% confidence interval (CI), 0.63-0.99; PR: HR, 0.69; 95% CI, 0.51-0.94) together with FIGO stage, residual tumor after primary surgery, age at diagnosis and other histological types vs. serous adenocarcinoma. The histological grade of tumor was found to have no independent prognostic value. The prognostic value of ER and PR was found additive with a HR for patients with high ER and PR expression of 0.48 (95% CI, 0.31-0.74) compared to patients with <10% expression for both receptors. In conclusion, our results predict that an elevated expression of ER and PR, alone and in combination, point to a favorable outcome for patients with OC. PMID- 17914555 TI - Gene expression profile of oral squamous cell carcinomas from Sri Lankan betel quid users. AB - Oral squamous cell carcinoma (OSCC) is one of the major health problems in Sri Lanka and the disease is associated with the habit of Betel Quid (BQ) chewing. Using 35k oligo microarrays, we analyzed the gene expression profile of 15 Sri Lankan patients diagnosed with OSCCs and pair-wised normal controls and correlated the findings with the clinicopathological data. Following the recording of the scanned array images and data analysis, results for selected candidate genes were verified using QRT-PCR. Upon analysis, a total of 263 genes [71 (27%) of unknown functions previously not reported in OSCCs and 192 (73%) of known functions] were found as differentially expressed between tumors and controls. For the genes with known functions, 66 (34%; such as COL4A1, MMP1, MMP3, PLAU, SPARC and KRT19) were previously reported in OSCC and for the remaining 126 (66%; such as CD47, APOL3, RRAGC, BPIL1 and AZGP1) this is the first report in OSCCs. Hierarchical clustering of the differentially expressed 263 genes grouped the samples into several clusters with the larger one being dominated by tumors of stage 3 and 4. Two cases (a verrucous SCC and an advanced SCC), did not cluster with any of the other samples. We found two main biological pathways (cell communication and integrin-mediated cell adhesion) and 5 gene ontology categories (transcription regulator activity, structural molecule activity, intracellular signaling, cytoskeleton and signal transduction) of relevance to the OSCCs examined. Results from the QRT-PCR verified the results from the microarray experiment. This study provides valuable information on gene expression profile of OSCCs of habitual users of BQ from Sri Lanka. Of particular interest were the list of genes of known and unknown functions and the two biological pathways that we suggest as candidate genes in oral cancers associated with BQ chewing in Southeast Asia, in particular Sri Lanka. The suggested candidate genes might be used as molecular biomarkers in the early detection of the alarming problem of OSCCs in Southeast Asia in association with BQ use. These findings provide valuable information that might help in the selection of possible biomarkers that can be used in early detection of the alarming problem of oral cancer in Southeast Asia. PMID- 17914556 TI - Investigation of optimal schedule of concurrent radiotherapy with S-1 for oral squamous cell carcinoma. AB - In the present study, we examined the appropriate schedule of S-1 medication in the combination with radiation by investigating the safety, the clinical efficacy, and antitumor effects on tumors in nude mice. In the patients with oral squamous cell carcinoma (OSCC), S-1 was given orally according to a 4-week application followed by 2-week rest regimen (4-week regimen), or a 2-week application followed by a 1-week rest regimen (2-week regimen). Radiation was given (2 Gy/day; 5 days/week) for a total of 60 Gy. In nude mouse models, human oral cancer cell lines were used as subcutaneous xenografts in nude mice. The mice were treated by S-1 (10 mg/kg) and radiation (1 Gy) with a 4-week regimen or a 2-week regimen. Apoptotic cells were detected by TUNEL method. In the patients with OSCC, the response rate with the 4-week regimen was 100% and the response rate with the 2-week regimen was 92.3%. However, a high frequency of adverse effect was found in the 4-week regimen when compared to the 2-week regimen. Grade 3 toxicity of leukopenia, neutropenia and stomatitis were seen in 3 cases, grade 3 toxicity of anorexia and nausea were seen in 2 cases, and grade 3 toxicity of decrease of hemoglobin level, heartburn/dyspepsia and increase of bilirubin level were seen in a case of the 4-week regimen. On the other hand, grade 3 toxicity of stomatitis, anorexia, nausea, heartburn/dyspepsia and increase of bilirubin level were seen in a case of the 2-week regimen. In nude mouse models, the 2-week regimen was more effective than the 4-week regimen. In addition, significant increase in the percentage of apoptotic cells was observed in the tumors treated with the 4-week regimen when compared with the tumors treated with the 2-week regimen. No loss of body weight was observed in mice treated with the 2-week regimen during the experimental period. These results suggested that the 2-week regimen might reduce adverse effects, and enhance therapeutic effects compared to the 4-week regimen. Briefly, this 2-week regimen may be a useful concurrent chemo radiotherapy improving the quality of life (QOL) of patients with OSCC. PMID- 17914557 TI - In vitro antitumor cytotoxic T lymphocyte response induced by dendritic cells transduced with DeltaNp73alpha recombinant adenovirus. AB - DeltaNp73alpha, the N-terminal truncated form of p73alpha is a candidate tumor antigen because of its selective expression in many human cancers and lack of expression in normal tissues. Therefore, we investigated the effects of dendritic cells infected with adenoviral DeltaNp73alpha (DNp73alpha) on breaking immune tolerance and induction of immunity against DNp73alpha-expressing (A549 lung cancer, K-562 leukemia) and non-expressing (MCF-7 breast cancer) cell lines. Immature dendritic cells generated in the presence of interleukin-4 and granulocyte/macrophage colony-stimulating factor from a human umbilical cord blood were transduced with a recombinant adenoviral (Ad) vector encoding full length human DNp73alpha cDNA (Ad-DNp73alpha) or a control vector Ad-EGFP, using the centrifugal force method. Induction of DNp73alpha-specific CTL response was evaluated by a cytotoxic assay against the three human tumor cell lines with different DNp73alpha expression levels. The viability and activation status of transduced dendritic cells were assessed by flow cytometry. The dendrocyte/Ad DNp73alpha-activated cytotoxic T lymphocytes showed significantly higher cytotoxicity against the cell lines A549/DNp73alpha, K-562 that expressed DNp73alpha than the DNp73alpha-null MCF-7 cells. The DCs/Ad-DNp73alpha showed higher survival rates than the DCs/Ad-EGFP or untransduced DCs, presumably due to the inhibition of cell death. These findings, with potential applications for immunotherapy, demonstrate that dendrocytes transduced with Ad-DNp73alpha can induce specific and sustained T cell responses against tumors expressing this variant p53-related gene. PMID- 17914558 TI - Involvement of the serrated neoplasia pathway in inflammatory bowel disease related colorectal oncogenesis. AB - The purpose of this study is to identify colorectal serrated lesions in the inflammatory mucosa of inflammatory bowel disease (IBD), to characterize their molecular status based on BRAF and KRAS mutations, mismatch-repair (MMR) deficiency and microsatellite instability (MSI), and to verify that these molecular alterations are specific to the 'serrated neoplasia pathway' in IBD. Neoplastic lesions from 36 patients with IBD were reviewed retrospectively, including 13 adenocarcinomas (1 mucinous and 12 conventional), 28 dysplasias [1 traditional serrated adenoma (TSA) and 27 conventional adenomas] and 1 hyperplastic polyp (HP). Both the HP and TSA exhibited the V600E BRAF mutation without MSI or MMR deficiency. The mucinous adenocarcinoma, close to the TSA, exhibited the BRAF mutation and MSI with loss of hMLH1. No KRAS mutations were found in these 3 lesions, and no BRAF mutations were found in the conventional ones. Serrated lesions exist in the inflammatory mucosa of IBD and are associated with a characteristic molecular profile, i.e. the appearance of the BRAF mutation as early as the hyperplastic polyp stage followed by MSI at the carcinoma stage. We therefore identified the serrated neoplasia pathway in IBD-related colorectal oncogenesis. PMID- 17914559 TI - Multicolor interphase cytogenetics for the study of plasma cell dyscrasias. AB - Specific chromosomal abnormalities such as chromosome 13 deletions and some translocations affecting the immunoglobulin heavy chain (IGH) gene, namely t(4;14)(p16;q32) and t(14;16)(q32;q23) have been associated with an adverse prognosis in multiple myeloma. Conventional cytogenetic techniques fail to detect these aberrations in the majority of cases. Thus, we have developed a novel set of interphase fluorescence in situ hybridization (I-FISH) assays targeting those regions frequently lost on chromosome 13 as well as those oncogenes most recurrently involved in translocations with the IGH locus in multiple myeloma, i.e. IRTA1/2 (1q21), FGFR3/MMSET (4p16), CCND3 (6p21), IRF4 (6p25), CCND1 (11q13), MAF (16q23), and MAFB (20q12). The probes were combined in a multicolor fashion to develop novel multicolor I-FISH (MI-FISH) assays, whose validity and applicability was evaluated in negative controls and in a series of 13 plasma cell neoplasias. Additionally, a combination of the novel MI-FISH assays with staining for the plasma cell-specific antigen VS38c by means of multicolor FICTION (M-FICTION, fluorescence immunophenotyping and interphase cytogenetics as a tool for the investigation of neoplasms) allowed us to selectively analyze the plasma cell compartment, and thereby to increase the assay sensitivity. PMID- 17914560 TI - Dual role of CCL3/CCR1 in oral squamous cell carcinoma: implications in tumor metastasis and local host defense. AB - Chemokines are small chemotactic cytokines that can induce the migration of leukocytes, activate inflammatory/immune responses and have recently been implicated in the regulation of tumor growth and organ-specific spread. In this setting, the macrophage inflammatory protein-1alpha (CCL3) chemokine displays a diversity of roles that may contribute to the directional migration of squamous cells into cervical lymph nodes or to the defense against tumor initiation and progression. Thus, the aim of this study was to determine, for the first time, the expression of CCL3 and their receptors, CCR1 and CCR5, by real-time polymerase chain reaction in samples obtained from oral squamous cell carcinoma (OSCC) and healthy gingival tissue (control). In addition, we investigated the immunoexpression of these molecules in neoplastic cells (parenchyma), inflammatory/immune cells (stroma) in primary OSCC and in metastatic and non metastatic lymph node tissues. The relationship of CCL3/CCR1 with survival data was also evaluated. The analysis of mRNA expression revealed a significantly higher expression of CCL3 and CCR1 in OSCC compared with the controls (P<0.05). The expression of CCR5 was not different in the two groups. The percentages of CCL3+ and CCR1+ cells were observed to be similar in parenchyma and stroma in the OSCC without lymph node metastasis when compared with OSCC with lymph node metastasis (P>0.05). However, we observed the density of CCL3+ nodal cells to be significantly higher in metastatic lymph nodes when compared with non-metastatic lymph nodes in the same patients (P<0.05). Considering CCL3 in stroma, the mean survival rate for patients with high CCL3+ cell percentage was better than for those with low CCL3+ cell percentage. Our findings suggest that the CCL3/CCR1 axis may have a role in the spread of tumoral cells to the lymph nodes and also in the local host defense against the tumor. PMID- 17914561 TI - Utility of tumour-infiltrating CD25+FOXP3+ regulatory T cell evaluation in predicting local recurrence in vertical growth phase cutaneous melanoma. AB - Tumour-infiltrating lymphocytes (TILs) represent the local immune response to cancer, however, their correlation with tumour behaviour is not unanimously considered in the literature. Most studies have not characterized TILs, that are known to comprise distinct subsets, bearing different roles in the complex tumour microenvironment. Characterization of patient lymphocytes has been mainly performed in peripheral blood, that is not always representative of the local immune status. Only few investigations have been performed at the tissue level in cancer, including melanoma. TILs encompass different populations of effector and regulatory T cells (Tregs), and the relevance of the latter in tumour progression is widely accepted. The transcription factor gene product FOXP3 is considered the most reliable marker of Tregs. However, it has not been extensively evaluated in primary cutaneous melanoma. We analyzed 66 vertical growth phase primary cutaneous melanomas, aiming at finding differences in TIL subsets between two groups of cases, that behaved differently in terms of local recurrence. In our study, the percentage of Tregs, as characterized by CD25 and FOXP3 expression, both among tumour cells, inside tumour parenchyma and at its periphery, and among TILs, at the tumour-stroma boundary, was significantly higher in cases that recurred than in those that did not (p=0.00065; p=0.00014; p<0.00001, respectively). TIL characterization by immunohistochemistry in melanoma diagnostic reports, could add further information. The analysis of a larger series of patients and correlation with other clinical parameters, such as distant metastases and/or patient survival, are mandatory for validating its use as a prognostic indicator. PMID- 17914562 TI - Microsatellite instability and clinicopathological features in esophageal squamous cell cancer. AB - Since multiple genetic alterations are involved in the molecular pathogenesis of esophageal squamous cell cancer (ESCC), the role of microsatellite instability (MSI) in its carcinogenesis is not well defined. The reported frequency of MSI in ESCC ranges from 2 to 66.7% but the majority of the results are derived from relatively small studies. Therefore, we carried out a precise MSI analysis on a large number of ESCC samples to clarify the significance of MSI in the ESCC tumorigenesis. The MSI status of the DNA extracted from 62 ESCC samples and 62 counterpart-normal esophageal epitheliums were studied with five NCI panel markers and ten microsatellite markers located in 17q24-25. Forty-four paraffin embedded samples and 18 frozen samples from the ESCC patients who underwent surgery were studied. The MSI status was classified as MSS (microsatellite stable), MSI-L (low-level MSI; <30% of markers examined showed instability) and MSI-H (high-level MSI; >30% of markers reported instability). Among the 62 ESCC cases analyzed by the 15 microsatellite markers, 38 out of 62 cases (61.3%) showed MSS, 19 out of 62 cases (30.6%) showed MSI-L and 5 out of 62 cases (8.1%) showed MSI-H. Although the MSI status was not associated with the status of lymph node metastasis or a histological type of cancer, the depth of cancer invasion was significantly associated with the frequency of MSS status and the levels of MSI-L were inversely correlated with the depth of invasion (T1/T2 vs. T3/4; P=0.0007). However, MSI status was not associated with the prognosis of the ESCC patients. This is the first large scale MSI analysis of the ESCC in comparison with the clinicopathological features. Relatively high frequency of MSI-L was observed in ESCC and the frequency of MSI-L was inversely correlated with the depth of invasion. PMID- 17914563 TI - Decreased expression of hMLH1 correlates with reduced 5-fluorouracil-mediated apoptosis in colon cancer cells. AB - Patients with sporadic microsatellite instable colorectal cancers, in most of which the function of the hMLH1 mismatch repair gene is impaired, do not gain a survival benefit from 5-fluorouracil (5-FU)-based chemotherapy. However, the effect of hMLH1 on the cytotoxicity induced by 5-FU has not yet been sufficiently confirmed. In this study, we assessed the effect of hMLH1 on cytotoxicity and apoptosis induced by 5-FU using newly developed cell lines. We constructed two cell lines: SW480 (originally hMLH1-proficient), in which the expression of hMLH1 was reduced using a small interfering RNA (siRNA) technique, and HCT116 (originally hMLH1-deficient), in which the expression of hMLH1 can be regulated by doxycycline. Using these cell lines, a clonogenic survival assay, 4',6 diamidino-2-phenylindole (DAPI) staining and an Annexin-V assay were performed. Moreover, the incorporation of 5-FU into DNA was determined using tritium-labeled 5-FU. In both of our two cell lines, hMLH1-deficient cells exhibited approximately 2.4-fold clonal surviving fraction compared to hMLH1-proficient cells for 10 days after the administration of 5-FU. Additionally, hMLH1-deficient cells treated with 5-FU exhibited 34-45% less apoptosis than hMLH1-proficient cells according to the results of DAPI staining and Annexin-V assay. Furthermore, hMLH1-deficient cells treated with 5-FU exhibited an approximately 2-fold greater incorporation of 5-FU into DNA than control cells, suggesting that the recognition of 5-FU-incorporated DNA is impaired in hMLH1-deficient cells, resulting in reduced apoptosis. Our conclusions were that decreased expression of hMLH1 in colon cancer cells reduced the apoptosis induced by 5-FU, suggesting that hMLH1 is a key determinant of 5-FU chemosensitivity. PMID- 17914564 TI - Increased expression of matrix metalloproteinase 9 and tubulin-alpha in pulmonary sclerosing hemangioma. AB - Pulmonary sclerosing hemangioma (PSH) is relatively rare and is usually considered a benign tumor because of its slow growth and solitary characteristics. However, several cases with lymph node metastasis have been reported, and its pathogenesis has not been fully elucidated. Three sets of PSH specimens from the Korea Lung Tissue Bank, obtained with IRB approval, were analyzed through the construction of an oligo-microarray that contained about 32,000 genes. The resulting data were confirmed by real-time RT-PCR. Protein expression levels were checked by performing immunohistochemistry (IHC) and immunoblot analysis. In the 3 specimens of PSH tissues, 72 of the 32,000 genes were commonly found up-regulated and 290 were commonly found down-regulated as compared to non-tumor tissues from each patient. Paraffin-embedded tissues from 11 cases were used to confirm the expression of matrix metalloproteinase 9 (MMP 9) and tubulin-alpha proteins in the non-tumor and PSH tissues via IHC. In addition, the upregulation of protein expression was confirmed by immunoblot analysis. As expected, in all cases MMP-9 and tubulin-alpha were expressed at significantly higher levels in the PSH than in the non-tumor tissues. This is the first report on a study of the whole genome of PSH. Increased expression of MMP-9 could induce the metastatic ability of PSH and tubulin-alpha might be responsible for the sclerotic character of this disease. The results of this study will be useful in helping to understand and effectively manage patients suffering from PSH. PMID- 17914565 TI - Metallothionein is up-regulated under hypoxia and promotes the survival of human prostate cancer cells. AB - Tumor hypoxia is a common feature of several cancers, including prostate cancer, and is associated with tumor progression, acquisition of anti-apoptotic potential and therapeutic resistance. We explored hypoxia-inducible genes and examined the effect of knockdown of a target molecule with small interference RNA (siRNA) on the proliferation of human prostate cancer cells. Human prostate cancer cell lines (LNCaP and PC-3) were cultured in normoxia (21% O2) or hypoxia (0.5% O2). Hypoxia-inducible genes were identified by cDNA microarray analysis. Metallothionein (MT) expression was assessed by real-time RT-PCR, Western blot analysis and immunohistochemical staining. siRNA was transfected to knock down MT expression, and the cell cycle and apoptosis were evaluated by flow cytometry analysis. In cDNA microarray analysis, 22 genes (including MT) were up-regulated under hypoxia. MT-1X and MT-2A were up-regulated in real-time RT-PCR. In particular, MT-2A was increased 3-fold in LNCaP and 8-fold in PC-3. The siRNA-MT 2A treatment resulted in a 20% inhibition of cell growth and induced apoptosis in both LNCaP and PC-3. In human prostate tissue, intense staining of MT was observed in cancer cells and residual cancer cells after androgen ablation therapy, while normal tissue was only stained in patches. In conclusion, MT was up-regulated under hypoxia in prostate cancer cells and overexpressed in prostate cancer tissue and residual cancer cells after androgen ablation therapy. As down regulation of MT by siRNA inhibited cell growth and induced cell death, MT may be a new molecular target for the treatment of human prostate cancer. PMID- 17914566 TI - Clinical implication of vascular endothelial growth factor T-460C polymorphism in the risk and progression of prostate cancer. AB - Vascular endothelial growth factor (VEGF), one of the most potent angiogenic factors, is suggested to play a crucial role in tumor neovascularization and is associated with tumor progression and metastasis in prostate cancer. This study evaluated the significance of the VEGF T-460C polymorphism in the risk and the progression of prostate cancer. In a case-control experiment, 270 patients with prostate cancer and 252 male controls were investigated to assess the association of the VEGF T-460C polymorphism with the risk of prostate cancer. Prostate specific antigen (PSA) recurrence in 95 patients who underwent radical prostatectomy and survival in 99 patients with metastases at diagnosis were analyzed to evaluate the influence of the polymorphism in cancer progression. The CC and TC genotypes of the polymorphism were associated with significantly higher rates of PSA recurrence after radical prostatectomy than the TT genotype and were independent predictors of PSA recurrence (P=0.011) in a multivariate analysis. In contrast, metastatic prostate cancer patients with the TT genotype showed significantly worse survival as compared to the CC and TC genotypes. In a multivariate analysis, the TT genotype was an independent predictor of cancer specific survival (P=0.006). The VEGF T-460C polymorphism may have a substantial impact on both PSA recurrence after radical prostatectomy and survival in advanced prostate cancer. The molecular mechanisms of the polymorphism on the differing status in prostate cancer should be elucidated in further studies. PMID- 17914567 TI - 8-Nitroguanine as a potential biomarker for progression of malignant fibrous histiocytoma, a model of inflammation-related cancer. AB - Chronic inflammation is a critical component of carcinogenesis and tumor progression. Reactive nitrogen and oxygen species generated by inflammatory cells form mutagenic DNA lesions, such as 8-nitroguanine, which may play an integral role in inflammation-related carcinogenesis. Hypoxia-inducible factor (HIF) 1alpha has been established as a prognostic biomarker in various tumors, including malignant fibrous histiocytoma (MFH). The aim of this study was to evaluate the impact of 8-nitroguanine formation and HIF-1alpha expression on the prognosis of patients with inflammation-related cancer. Immunohistochemical analyses were employed to examine the distribution of 8-nitroguanine and HIF 1alpha, using clinical specimens from 36 patients with MFH as a model of inflammation-related cancer. 8-Nitroguanine formation was predominantly observed in the nuclei of tumor cells and inflammatory cells in tumor tissues, while HIF 1alpha was expressed in the cytoplasm and nuclei of tumor cells. Little or no immunoreactivity of 8-nitroguanine and HIF-1alpha was observed in adjacent non tumor tissues. Significantly higher levels of both 8-nitroguanine and HIF-1alpha were observed in the tissue specimens of deceased patients than in those of living subjects. Survival curves analyzed by the Kaplan-Meier method differed significantly between the high- and low-staining groups of 8-nitroguanine (p=0.00003) as well as HIF-1alpha (p=0.01104). These results suggest a significant role of the pathway of iNOS-dependent 8-nitroguanine formation via HIF-1alpha and NF-kappaB on the progression of inflammation-related cancer. In conclusion, 8-nitroguanine is an excellent candidate prognostic and predictive biomarker together with HIF-1alpha in inflammation-related tumor progression. PMID- 17914568 TI - Single nucleotide polymorphism in the RAD18 gene and risk of colorectal cancer in the Japanese population. AB - The RAD18 gene, located on the human chromosome 3p24-p25, plays a crucial role in post-replication repair (PRR) in various organisms from yeast to humans. In the human RAD18 gene, one coding single nucleotide polymorphism (SNP) at codon 302, encoding either arginine (Arg, CGA) or glutamine (Gln, CAA), was reported. Although the molecular function of the RAD18 protein came to be elucidated, the association between the RAD18 Arg302Gln polymorphism and the risk of human cancer development was not examined. Therefore, we investigated the relationship between the polymorphism and the development of human primary colorectal cancer (CRC). The Arg302Gln polymorphism in 100 patients with CRC and 200 healthy controls were genotyped by the polymerase chain reaction with confronting two-pair primer (PCR CTPP) assay. The Gln/Gln genotype was significantly more frequent in CRC (18.0%) than in the healthy controls (11.5%) (p=0.046). The increased risk was detected in CRC patients with the Gln/Gln genotype (Odds ratio [OR], 2.10; 95% confidence interval [CI], 1.00 to 4.40). When the relationship of the SNP with clinicopathological parameters of CRC was investigated, particularly in the well differentiated grade and in the lymph node metastasis (N1) CRC patients, significantly higher risks were detected (OR, 7.00; 95% CI, 1.19-41.1 and OR, 3.71; 95% CI, 1.30-10.6, respectively). These results suggested that the RAD18 Arg302Gln polymorphism is associated with the risk of CRC. This report provides evidence for an association between the RAD18 Arg302Gln polymorphism and human CRC risk. PMID- 17914569 TI - Epidemiological survey of oral lichen planus among HCV-infected inhabitants in a town in Hiroshima Prefecture in Japan from 2000 to 2003. AB - The objective of our study was to evaluate the natural history of oral lichen planus (OLP) and other extrahepatic manifestations in the inhabitants of an area in Japan that is hyperendemic for hepatitis C virus (HCV) infection. Over 4 years, 224 adult inhabitants with HCV infection were examined for OLP by a single oral surgeon. All subjects were interviewed regarding the natural history of other extrahepatic manifestations they had developed. The antibodies to HCV (anti HCV) and serum HCV RNA were determined. Anti-HCV were detected in sera from 224 subjects (100%); HCV RNA in 210 (93.8%). Of the 224, 88 had at least 1 oral examination for OLP during the 4-year period. In 2000, 2001, 2002 and 2003, OLP was observed in 8.5 (5/59), 14.8 (8/54), 20 (11/55) and 21.4% (12/56) of subjects, respectively. OLP prevalence increased as the subjects grew older. The incidence of OLP over the 4 years among all subjects with HCV infection was 17.0% (15/88, 2 men and 13 women). None experienced natural healing or the development of malignant transformations. Between 2000 and 2003, there was an increase in the prevalence of type 2 diabetes mellitus (DM), thyroid dysfunction, skin disease, renal disease and hypertension. Screening for extrahepatic manifestations should be conducted in patients with risk factors for HCV infection. PMID- 17914570 TI - Potential for molecular targeted therapy of HER-2/neu for invasive bladder cancer: examination of gene amplification by fluorescence in situ hybridization. AB - Analysis of HER-2/neu gene amplification by fluorescence in situ hybridization was performed in 40 patients with invasive bladder cancer in order to evaluate the potential for molecular targeted therapy of HER-2 as a tailor-made treatment for patients with invasive bladder cancer. This study included 40 patients seen at the Aichi Medical University Hospital from January 2001 to December 2004 and were pathologically diagnosed with invasive transitional cell carcinoma of the bladder (pT2-pT4). The PathVysion kit was used to evaluate the status of HER 2/neu gene amplification, and a signal ratio > or =2.0 was considered positive for HER-2/neu gene amplification. In primary foci 5 patients (12.5%) were positive for HER-2/neu gene amplification. According to the classification of grade and stage, no statistically significant difference was observed. Lymph node metastasis was found in 10 patients, and 3 patients (30%) were positive for HER 2/neu gene amplification. In the patients with HER-2/neu gene-amplified metastatic lymph nodes, primary foci were also positive for gene amplification, showing a statistically significant difference. This study indicates that 12.5% of patients with invasive bladder cancer may benefit from molecular targeted therapy of HER-2, and that molecular targeted therapy can be expected to be effective even for patients with lymph node metastases as long as their primary foci are positive for HER-2/neu gene amplification. PMID- 17914571 TI - Expression of AITR and AITR ligand in breast cancer patients. AB - Human activation-inducible TNF receptor (AITR) is a new member of the tumor necrosis factor family and expressed on peripheral blood mononuclear cells (PBMC) and leukocytes. Its ligand (AITRL) is expressed in endothelial cells. This study aimed to evaluate the presence and role of AITR and AITRL in patients with breast cancer. Expression of AITR and AITRL on PBMC and breast cancer cells was determined by flow cytometry, RT-PCR and fluorescence microscopy. Soluble (s) AITR and sAITRL were detected in serum of breast cancer patients by ELISA. AITR and AITRL were constitutively expressed in T cells, B cells, monocytes and monocyte-derived dendritic cells of breast cancer patients. AITRL was overexpressed in breast cancer cells. The levels of sAITRL were significantly increased in serum of breast cancer patients compared with the healthy control. This study suggests that AITR and AITRL may play an important role in tumor growth and survival in breast cancer. PMID- 17914572 TI - Establishment and characterization of a human cholangiocarcinoma cell line. AB - Cholangiocarcinoma (CC) is a rare malignant tumor arising from the biliary tract. The disease is notoriously difficult to diagnose and is usually fatal due to its late clinical presentation and the lack of effective non-surgical therapeutic strategies. To date, little is known about the cancer biology of the disease and the establishment and characterization of only a few CC cell lines have been reported. We report here the establishment of a new human cancer cell line, HKGZ CC, from a moderate to poorly differentiated intrahepatic bile duct carcinoma from a Chinese patient. Morphological characteristics, growth kinetics, ability to grow on anchorage-independent soft agar, tumorigenicity in nude mice and cytogenetic features of the cell line were investigated. Chromosome banding karyotype and comparative genomic hybridization analyses revealed chromosomal changes in 1pter-p31, 1q31-qter, 3q, 8q21-qter, 9pter-9q34, 10, 13q21-qter and X. This newly established cell line should serve as a useful model for studying the molecular pathogenesis of CC. PMID- 17914573 TI - Immunohistochemical expression of VEGF-A and its ligands in non-neoplastic lesions of the breast sampling-assisted by dynamic angiothermography. AB - The aim of this study was to investigate the expression of angiogenic markers, vascular endothelial growth factor A (VEGF-A) ligand and its receptors, VEGFR-1 and -2, in a series of biopsy-proven non-neoplastic lesions of the breast detected by dynamic angiothermography. We have also studied the vascular density demonstrated by CD31 immunoreactivity, in order to assess the potential of the imaging method to recognize lesions with an enhanced vascular network of clinical importance in routine breast examination. The lesions were classified as non proliferative, proliferative without atypia and proliferative with atypia. VEGF was diffusely expressed in the epithelial cells of proliferative lesions in almost all cases. Similarly, VEGFR-1 and -2 also exhibited epithelial positive reactions in the majority of cases. VEGF-A and its receptors were also present in blood vessels. CD31 showed an increase in vascular proliferation at the periphery of proliferative epithelial lesions, but not in non-proliferative lesions. Our results, showing marked expression of VEGF by the epithelial proliferative lesions and neoangiogenesis at their periphery, confirm that these lesions can be detected by dynamic angiothermography. PMID- 17914574 TI - c-Jun, a crucial molecule in metastasis of breast cancer and potential target for biotherapy. AB - Human breast cancer cell line SKBR3 expresses high level of the ErbB2 molecule, which has been associated with poor prognosis of breast cancer. Elevated ErbB2 functions as a transactivating co-receptor and promotes the formation of ErbB2 containing heterodimers, which are more mitogenic and transforming, and have a higher ligand affinity and signaling potency by virtue of the potent latent kinase activity of ErbB2. Interestingly, SKBR3 cells are non-tumorigenic in nude mice. By ectopic overexpression of c-Jun in SKBR3 cells, involvement of c-Jun in invasiveness and metastasis in vivo was investigated in this study. The critical role of c-Jun in the tumorigenesis and metastasis is demonstrated in nude mice. PMID- 17914575 TI - Loss of TP53-DNA interaction induced by p.C135R in lung cancer. AB - The p53 tumor suppressor gene (TP53; OMIM: 191170) plays an important role in tumorigenesis in lung epithelial cells. TP53 encodes a sequence-specific DNA binding protein that regulates transcription of several genes in response to DNA damage promoting cell cycle arrest, DNA repair or apoptosis. A mutation does not necessarily alter the protein function and since not all altered tumor protein p53 (TP53) conformations lead to the same biological properties, we studied Cys135Arg TP53 gene mutation in squamous cell type of non-small cell lung cancers (NSCLCs), by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and direct sequencing. Cys135Arg TP53 mutation, rare in databases (11/23544 in R11, IARC TP53 database), was detected. We chose p.C135R in order to examine DNA-TP53 interaction. A comparison with the wild-type after 1 nano-second molecular dynamic simulation analysis revealed a significant structural change (over 4A displacement) in the contact loop Lys-Ser-Val which lies upstream and next to the mutated site in the TP53, that sterically prevents its DNA-binding activity. Additionally, the mutation produced a change in the electrostatic potential surface of the protein in the same loop where the structural modification took place. To demonstrate the degree of loss of function, functional assays in yeast and bacteria with oligonucleotides for competitive electrophoretic mobility shift assays (EMSAs) were done proving that this mutation decreases TP53 ability to bind DNA of the TP53 response element from the human p21 gene. These results demonstrate that the amino acid change C135R in the human TP53 generates the loss of TP53 DNA-binding activity directly affecting its role as a transcription factor and suggests that this observation can explain part of the phenotype described in patients affected by this type of tumor. PMID- 17914576 TI - Influence of cytological stains on comparative genomic hybridization analysis for DNA extracted from cytological smears. AB - In the present study, we investigated the influence of cytological stains in analyzing DNA extracted from cytological slides by comparative genomic hybridization (CGH). Multiple imprint cytological slides were prepared for fresh frozen breast cancer tissue samples and the slides were stained by three staining methods for each sample. Under microscopic observation, cancer cells were selectively microdissected from the slides and forwarded to DNA extraction, whole genome amplification, and CGH analysis. CGH was successfully performed for all methylgreen-stained and May-Grunwald-Giemsa (MGG)-stained cytological smear slides, but for two Papanicolaou (PAP)-stained slides. The number of chromosomal imbalances detected were 5-10 in methylgreen-stained slides and 5-9 in MGG stained slides. The chromosomal imbalances resemble each other between methylgreen-stained and MGG-stained slides. The present study indicates that the MGG stain is preferred to the PAP stain for the purpose of cytogenetical analysis by CGH for DNA extracted from cytological smear slides. PMID- 17914577 TI - Detection of promoter hypermethylation in serum samples of cancer patients by methylation-specific polymerase chain reaction for tumour suppressor genes including RUNX3. AB - The purpose was to validate the use of RUNX3 as a potential biomarker for detection of cancer in serum samples and to determine its sensitivity alone and in combination with p16, RASSF1A and CDH1 using methylation-specific polymerase chain reaction (MSP). We examined the promoter methylation status of RUNX3, p16, RASSF1A and CDH1 by MSP using the serum of 70 metastatic breast, non-small cell lung, gastric, pancreatic, colorectal or hepatocellular carcinomas. The DNA from 10 healthy serum controls was used to determine the specificity of methylation. According to our results, promoter hyper-methylation of RUNX3 was detected in the serum of 44 patients comprising breast 9/19 (47%), non-small cell lung 11/20 (55%), gastric 4/4 (100%), pancreatic 2/2 (100%), colorectal 11/17 (65%) and liver 7/8 (88%) carcinomas. Comparative figures for the other genes were as follows: p16 - 39/70 (7/19, 10/20, 2/4, 0/2, 12/17, 8/8); RASSF1A - 24/70 (8/19, 6/20, 1/4, 1/2, 4/17, 4/8); CDH1 - 10/70 (0/19, 4/20, 1/4, 1/2, 3/17, 1/8). Using a panel of four genes, hypermethylation of one or more genes was found in 62/70 samples (15/19, 19/20, 4/4, 2/2, 14/17, 8/8). A panel of three genes omitting RUNX3 detected hyper-methylation in only 50/70 samples. No methylation was detected in the 10 healthy serum controls. Thus, RUNX3 can be detected in the serum of a high proportion of advanced cancers. This suggests that serum hypermethylation of RUNX3 is at least as, or possibly more sensitive a marker, than other tumor suppressor genes currently under investigation. Inclusion of RUNX3 in gene panels can potentially increase the sensitivity of such panels for serum diagnosis of malignancies and warrants further study. PMID- 17914578 TI - New peptides of the polycomb group protein enhancer of zeste homolog 2 with the potential to induce cancer-reactive cytotoxic T lymphocytes in human leukocyte antigen-A2+ prostate cancer patients. AB - The polycomb group protein enhancer of zeste homolog 2 (EZH2) is linked to aggressive prostate cancer and could be an appropriate target in specific immunotherapy. In this study, we attempted to identify EZH2-derived peptides that have the potential to generate cancer-reactive cytotoxic T lymphocytes (CTLs) in human leukocyte antigen (HLA)-A2+ prostate cancer patients. Twelve EZH2-derived peptides were prepared based on the HLA-A2 binding motif. These peptide candidates were screened first by their ability to be recognized by immunoglobulin G (IgG), and then by their ability to induce peptide-specific cytotoxic T lymphocytes (CTLs). As a result, five EZH2 peptides recognized by IgG (EZH2 120-128, EZH2 165-174, EZH2 569-577, EZH2 665-674, and EZH2 699-708) were frequently detected in the plasma of prostate cancer patients. Among them, the EZH2 120-128 and EZH2 165-174 peptides effectively induced HLA-A2-restricted and cancer-reactive CTLs from prostate cancer patients. The cytotoxicity was mainly dependent on EZH2 peptide-specific and HLA-A2-restricted CD8+ T cells. These results indicate that these EZH2 120-128 and EZH2 165-174 peptides could be promising candidates in peptide-based immunotherapy for HLA-A2+ prostate cancer patients. PMID- 17914579 TI - Glycosylation modulates TRAIL-R1/death receptor 4 protein: different regulations of two pro-apoptotic receptors for TRAIL by tunicamycin. AB - Death receptor 4 (DR4) is a receptor of the antitumor death ligand, TNF-related apoptosis-inducing ligand (TRAIL), and is considered a promising molecular target for cancer therapy. Here, we show a novel regulation of DR4 protein. Tunicamycin treatment, which is an inducer of endoplasmic reticulum (ER)-stress, generated a lower molecular-weight pattern of DR4, but not DR5 protein in prostate cancer DU145 and PC3 cells. Thus, we termed the small form of DR4 protein, DR4-Small (DR4-S) and the large form, DR4-Large (DR4-L). Using DR4 siRNA, we confirmed that DR4-S also stands for DR4 protein. Other ER-stress inducers, brefeldin A and thapsigargin did not generate DR4-S. On the other hand, these ER-stress inducers increased DR5 protein. Tunicamycin induces ER-stress following the inhibition of N-linked glycosylation. Thus, we examined DR4 protein in cell lysates treated with glycosydase. Glycosydase treatments generated DR4-S protein, similar to tunicamycin. These results indicate that tunicamycin regulates DR4 protein size via inhibition of glycosylation. PMID- 17914580 TI - Predicting the tumor response to radiotherapy using microarray analysis (Review). AB - Predicting the tumor response to radiotherapy is one of the major goals of human cancer treatment. Identification of the genes that are differentially expressed between radiosensitive and radioresistant cancers by global gene analysis may provide new insights into the mechanisms underlying clinical radioresistance and improve the efficacy of radiotherapy. In this study, we reviewed the published reports identifying sets of discriminating genes using microarray analysis that can be used for characterization and the prediction of response to radiotherapy in human cancers. These reports indicate that many of the identified genes were associated with DNA-repair, apoptosis, growth factor, signal transduction, cell cycle and cell adhesion. Several genes were found to be predictors of the radiation response with various cancers and certain sets of identified genes were also found to predict the radiation response by using clustering analysis. Global gene expression profiling of responders and non-responders can be useful in predicting responses to radiotherapy and may also provide insights into the development of individualized therapies and novel therapeutic targets. PMID- 17914581 TI - Adjuvant chemotherapy with carboplatin and taxane compared with single drug carboplatin in early stage epithelial ovarian carcinoma. AB - The objective of the present study was to compare recurrence-free survival (RFS) in early stages (FIGO stages I-II) of epithelial ovarian cancer after adjuvant chemotherapy with carboplatin and a taxane (113 patients) and with carboplatin alone (27 patients). The distribution of clinical and pathological prognostic factors as well as type of primary surgery were comparable in the two groups. Recurrence rate was 21% and RFS was 79% in the series of patients treated with taxane-based chemotherapy and 19% and 81%, respectively, in the series of patients who received single drug carboplatin. Thus, no significant differences were recorded. The major toxicities in the present study were myelosuppression (46%) and neuro-toxicity (26%). Neuro-toxicity was more frequently (P=0.007) recorded and of higher grade (P=0.011) for patients in the carboplatin-taxane series compared with patients in the carboplatin series. RFS for patients in FIGO stage I was 85% and for patients in FIGO-stage II only 47%. In a multivariate logistic regression analysis of predictive factors for tumor recurrence in the complete series (n=140) the FIGO stage was the only independent and significant (P=0.0006) predictive factor with an odds ratio of 6.4 (95% CI: 2.2-18.9) for stage II versus IA-C. Age, tumor grade and type of adjuvant chemotherapy (+/- taxane) were not significant predictive factors. In the present study, although based on a limited number of patients, we could not find any improvement in recurrence rate or recurrence-free survival for patients treated with a carboplatin-taxane combination regimen compared with patients treated with carboplatin monotherapy. The spectrum of side effects was also in favor of the monotherapy regimen. Further, larger randomized studies are needed to give a final and fully conclusive answer to this question. PMID- 17914582 TI - Antitumor and anti-invasive effects of diverse new macrocyclic lactones, alkylolides and alkenylolides, and their enhancement by hyperthermia. AB - Alkylolides and alkenylolides of 198-254 Da such as hexadecan-16-olide and 9 hexadecen-16-olide were chemically synthesized in the present study as new macrocyclic lactones that are structurally different from widespread natural macrocyclic lactones including bryostatin (887 Da) and rhizoxin (613 Da), and were investigated for antitumor activity to Ehrlich ascites tumor cells by mitochondrial dehydroganase-based WST-1 assay and dye-exclusion assay. Of the alkylolides having 12, 15 or 16 carbon-atoms (D12:0, P15:0 or H16:0) and alkenylolides having 15 or 16 carbon-atoms with a double bond (P15:1 or H16:1), H16:0 was the most carcinostatic when administered at 37 degrees C for 20 h, with cell deformation and microvillus disappearance as detected by scanning electron microscopy. The carcinostatic activity was increased markedly for H16:0 and P15:0 when the administration period was prolonged to 72 h, but was not enhanced by intramolecular introduction of a double bond for P15:1 or H16:1. Hyperthermia at 42 degrees C for 30 min additively intensified the carcinostatic activity for H16:0 and P15:0, but scarcely for D12:0, and intensified the alkenyloides P15:1 and H16:1 only upon the subsequent 72-h treatment. Invasion of human fibrosarcoma HT-1080 cells through the reconstituted basement membrane was inhibited by alkyl- and alkenylolides even after the short-term exposure at 25 microM for 3 h without diminishing the cell viability. H16:0 also exhibited the most inhibitory activity to tumor invasion in addition to the highest carcinostatic activity. Both inhibitions were promoted by combination with hyperthermia. Thus diverse alkyl /alkenylolides, may be potent multi-applicable anticancer agents in terms of either dual inhibitory activities against both tumor progression and invasion or hyperthermia-combined therapy. PMID- 17914583 TI - Sulforaphane induces cell cycle arrest and apoptosis in murine osteosarcoma cells in vitro and inhibits tumor growth in vivo. AB - Sulforaphane (SFN), a naturally occurring isothiocyanate, is an attractive agent due to its potent anticancer effects. SFN suppresses the proliferation of various cancer cells in vitro and in vivo. In this study, we report that SFN inhibited the proliferation of cultured murine osteosarcoma LM8 cells. Twenty micromolar SFN completely inhibited the growth of LM8 cells and caused G2/M-phase arrest. SFN induced the expression of p21(WAF1/CIP1) protein causing the cell cycle arrest in a dose-dependent manner. SFN induced apoptosis which was characterized by the appearance of cells with sub-G1 DNA content and the cleavage and activation of caspase-3. We showed that SFN induced the growth arrest and up regulated the expression of p21(WAF1/CIP1) protein in a p53-independent manner in human osteosarcoma MG63 cells. We found that intraperitoneal administration of SFN (1 or 2 mg, 5 times/week) significantly inhibited the growth of LM8 xenografts to <30% of the controls in a preclinical animal model without causing any toxicity. In osteosarcoma cells, our findings provide in vivo evidence for the efficacy of SFN against the advanced growth of tumor. We showed that SFN induces cell cycle arrest and apoptosis in osteosarcoma cells and inhibits tumor xenograft growth. Furthermore, SFN is a potent inducer of p21(WAF1/CIP1) in osteosarcoma cells. These results raise the possibility that SFN may be a promising candidate for molecular-targeting chemotherapy against osteosarcoma. PMID- 17914584 TI - Resveratrol induces pro-apoptotic endoplasmic reticulum stress in human colon cancer cells. AB - Resveratrol (3,4',5 tri-hydroxystilbene), a naturally occurring polyphenolic compound highly enriched in grapes and red wine, has been shown to induce anti proliferation and apoptosis of human cancer cell lines. Resveratrol-induced dose dependent apoptotic cell death in colon carcinoma cells, was measured by FACS analysis. Treatment of HT29 human colon carcinoma cells with resveratrol was found to induce a number of signature ER stress markers; phosphorylation of eukaryotic initiation factor-2alpha (eIF-2alpha), ER stress-specific XBP1 splicing and CCAAT/enhancer-binding protein-homologous protein (CHOP). In addition, resveratrol induced up-regulation of glucose-regulated protein (GRP) 78, suggesting the induction of ER stress. Furthermore, the inhibition of caspase 4 activity by z-LEVD-fmk significantly reduced resveratrol-induced apoptosis. Taken together, the present study therefore provides strong evidence to support an important role of ER stress response in mediating the resveratrol-induced apoptosis. PMID- 17914585 TI - Comparative study of the effects of percutaneous ethanol injection and radiofrequency ablation in cases treated with a straight or expandable electrode. AB - Radiofrequency ablation (RFA) has become mainstream among non-surgical treatment modalities in clinical settings for the treatment of hepatocellular carcinoma. We have previously described the novel combination therapy of percutaneous ethanol injection and RFA (PEI-RFA) and reported that this combination therapy was more effective than RFA alone in terms of the induced volume of coagulated necrosis and the energy requirement for the treatment. RFA instruments are mainly divided into two types according to the electrode used, either the straight or expandable type electrode. Although PEI-RFA can be performed by either of the electrodes, there may be some important differences in PEI-RFA according to the type of electrode used. In the present study, the effect of using the straight or expandable electrode in PEI-RFA was evaluated by analyzing the ablation time, volume of coagulated necrosis, the energy requirement for ablation and the amount of injected ethanol into HCC. The comparative study showed that ablation time, total energy requirement and per unit volume of energy requirement for whole and marginal coagulated necrosis were significantly smaller in the group treated with the expandable electrode (E group) than those in the group treated with the straight electrode (S group). The volume of coagulated necrosis was similar between these groups. In group E, the amount of injected ethanol showed a positive correlation with the volume of coagulated necrosis and the size of the tumors. These results suggest that prior injection of ethanol works mainly by shortening the time and energy requirement for ablation in the time-lag PEI-RFA using the expandable electrode. Thus, prior injection of ethanol before RFA may make RFA treatment less invasive in the time-lag PEI-RFA using the expandable electrode as previously shown HCC cases treated with straight electrode. PMID- 17914586 TI - Mechanisms of anti-proliferative effect of JTE-522, a selective cyclooxygenase-2 inhibitor, on human liver cancer cells. AB - Selective cyclooxygenase-2 (COX-2) inhibitors have been demonstrated to inhibit the proliferation of a variety of cancer cells including hepatocellular carcinoma (HCC). We sought to explore the mechanisms by which JTE-522, a selective COX-2 inhibitor, suppressed the growth of human HCC cells. HCC cells (HepG2, HLF, huH1, Huh7, and PLC/PRF/5 cells) did not express COX-2 at either the mRNA or protein level. Prostaglandin E2 (PGE2) levels in medium were not significantly modulated by the JTE-522 treatment. However, MTT assays disclosed that escalating doses (100 nM to 100 microM) of JTE-522 significantly inhibited the growth of all HCC cells in a dose- and time-dependent manner. JTE-522 induced cell cycle arrest at the G1 phase, which was in part mediated by downregulation of cyclin E. Hallmarks of apoptosis, including the sub-G1 fraction by flow cytometric analysis and nuclear fragmentation by nuclear staining, were not significantly induced after the JTE-522 treatment. In addition, JTE-522 enhanced the expression of peroxisome proliferator-activated receptor (PPAR)-gamma protein in HepG2 and PLC/PRF/5 cells. Our data demonstrate that JTE-522 inhibited the growth of HCC cells in a COX-2-independent manner, and that the growth inhibition was in part mediated by the cell cycle arrest and the upregulation of PPAR-gamma protein. PMID- 17914587 TI - Isoprenoid-independent pathway is involved in apoptosis induced by risedronate, a bisphosphonate, in which Bim plays a critical role in breast cancer cell line MCF 7. AB - Bisphosphonates cause apoptosis to various types of cancer cells including breast cancer. Inhibition of the mevalonate pathway was reported to be involved in the apoptosis induced by bisphosphonates, but its precise mechanism has not been unveiled. In the present study, we investigated the molecular mechanism of risedronate, a bisphosphonate, in the apoptosis of the breast cancer cell line MCF-7 in comparison with that of cerivastatin, an HMG CoA reductase inhibitor (statin), since statin has been known to induce apoptosis through an isoprenoid dependent pathway in these cells. We found that i) risedronate induced MCF-7 cells into apoptosis in a manner similar to cerivastatin with the activation of caspase-9 followed by caspase-6 and -7, that ii) bisphosphonate-induced apoptosis was significantly, but not fully, recovered by the addition of GGOH, an isoprenoid, which completely rescued in case of cerivastatin-induced apoptosis, that iii) risedronate induced G2 arrest with the induction of Bim (BH3-only protein), but that statin induced G1 arrest without it, and that iv) the down regulation of Bim protein by siRNA significantly attenuated the risedronate induced apoptosis. These data clearly indicate that both isoprenoid-dependent and -independent pathways might be involved in the apoptosis induced by bisphosphonate, and Bim might be a critical component for the isoprenoid independent apoptotic pathway. PMID- 17914588 TI - Calculated and simulated effects of heterogeneous dose distributions in radiotherapy using the dose volume inhomogeneity corrected biological equivalent dose formula with special reference to prostate cancer. AB - The aim of this study was to evaluate and estimate the impact on the biological effective dose (BED) of irradiation delivered to a tumour during high dose rate brachytherapy with a heterogeneous dose distribution in the target volume. The calculation of BED in combination with the critical-voxel model and the LQ (linear quadratic) model was used to evaluate the effect of different combinations of heterogeneous dose distribution. The model is called the dose volume inhomogeneity corrected BED (DVIC-BED). Different random and non-random combinations of radiobiological parameters were used to test the model. A simulated clinical treatment of two 10 Gy fractions of brachytherapy was used. In the simulations 0-100% of the target volume was simulated to receive only 80% of the total dose. Different alpha/beta ratios as well as a different alpha value were used. A drastic effect on the outcome was observed especially when the ratio was low and the alpha value was high. The clinical effect is amplified when the tumour is considered to have a step dose respond curve. A 5 Gy decrease in dose corresponds to a reduction in clinical or chemical control < or =10-25% in the interval 65-85 Gy. Random production of different values has basically the same effect as above. The result stresses the importance to have control of the dose and the target volume during brachytherapy of prostate cancer. This is even more important when monotherapy with high dose rate brachytherapy is used and with a low alpha/beta ratio. The advantage of using this formula is that it is based on the LQ/BED formula and that different treatments with different fractions and treatments can be summated independently of the homogeneity of the dose distribution. PMID- 17914589 TI - Apoptotic effects of signal transduction inhibitors on human tumor cells with different PTEN expression. AB - An important mechanism of antitumoral targeted therapies is the induction of apoptosis in tumor cells. Tamoxifen and trastuzumab (Herceptin), respectively, are able to trigger apoptosis in human breast cancer cells. But, frequently altered apoptotic signal cascades, for instance through PTEN mutations, help tumor cells to escape antitumoral therapy. We studied to what extent the apoptotic effect of signal-transduction inhibitors is dependent on PTEN expression. PTEN expression was analysed by Western blot analysis in tumor cell lines of the breast (BT-474, MCF-7, MDA-MB-231), ovary (BG-1, SK-OV-3) and endometrium (Ishikawa, HEC-1A). Apoptotic effects of tamoxifen, trastuzumab, ZD1839 (Iressa) and different mitogen-activated protein kinase (MAP) inhibitors were measured after 24 h of treatment. Cellular apoptosis was determined by the detection of cytoplasmic histone-DNA complexes. The tested tumor cell lines exhibited a different PTEN expression, ranging from a high expression (ovarian cancer cell line BG-1 and BT-474 breast cancer cells) to a total absence of PTEN expression (endometrial Ishikawa cells). The apoptotic effect of receptor targeting drugs (tamoxifen, trastuzumab, ZD1839) was dependent both on receptor expression and PTEN expression. When cells were treated with MAPK inhibitors, no correlation between PTEN expression and the apoptosis rate was observed. Our data underline the importance of PTEN expression regarding the induction of apoptosis through various targeted therapies. PMID- 17914590 TI - Metastasis-associated protein 1 inhibits p53-induced apoptosis. AB - Metastasis-associated protein 1 (MTA1) is highly upregulated in cancer cells with metastatic potential; however, the molecular mechanism by which MTA1 increases the metastatic potential of cancer cells is far from clear. We characterized the functional consequences of MTA1 overexpression on p53-induced apoptosis of cancer cells. MTA1 was associated with p53 in a co-immunoprecipitation assay. MTA1 also had deacetylation activity on p53 in human non-small cell lung cancer cells H1299 and human hepatoma cells SK-Hep1. MTA1 attenuated the transactivation and p21 induction by p53. Moreover, MTA1 expression decreased p53-mediated apoptosis. These results indicate that MTA1 inhibits p53-induced apoptosis by deacetylation of p53, which might be related to the increased metastatic potential of cancer cells with high MTA1 expression. PMID- 17914591 TI - Internalization of Mycobacterium bovis, Bacillus Calmette Guerin, by bladder cancer cells is cytotoxic. AB - Instillation of Bacillus Calmette Guerin (BCG) into the bladder is the standard treatment for superficial bladder cancer. It leads to a local inflammatory response due to the release of cytokines and influx of immune cells to the tumor site. Although the presence of an intact immune system is an essential criterion for successful therapy, attachment of the bacteria to the bladder urothelial is just as important. The purpose of our study is to determine the role of bacterial internalization by epithelial cells. Transfection of the alpha5 integrin gene into the BCG unresponsive bladder cancer cell line, RT4, caused an increase in bacterial uptake and also increased cell death. Treatment of cells with cycloheximide did not prevent bacterial internalization but blocked its cytotoxic effect suggesting that unlike cell death, the process of bacterial internalization does not require new protein synthesis. Our data also show that the bacteria secretory products can prevent its own internalization. The extract prepared from lyophilized BCG altered the phosphorylation status of the focal adhesion kinase which is responsible for cellular endocytosis. Therefore, bacterial phosphatases may be present in the bacterial extract. Their activity may inhibit BCG internalization. Thus washing the reconstituted bacteria to remove the enzymes before instillation into the bladder might improve the therapeutic outcome of intravesical BCG therapy. PMID- 17914592 TI - Gefitinib and bicalutamide show synergistic effects in primary cultures of prostate cancer derived from androgen-dependent naive patients. AB - We previously demonstrated that the inhibition of the epidermal growth factor receptor (EGFR) signalling affects the endocrine therapy responses of prostate cancer (PCa) cells and that bicalutamide (BCLT) is able to reinforce PI3K activity through mechanisms involving PTEN decrement and EGFR and Her2 activities. The aim of this study was to evaluate if the hormonal therapy with BCLT can affect the EGFR-targeted therapy using primary cultures obtained from 22 human PCa tissues harvested after radical prostatectomy (RP) in patients who received (n=10) BCLT and those that did not (n=12) as neoadjuvant hormone therapy (NHT). We demonstrated that cultures derived from PCa tissues harvested after NHT presented significantly higher EGFR and Her2 levels compared to cultures derived from control patients. However, cultures derived from patients with NHT were less sensitive to gefitinib when compared to cultures derived from control patients. Conversely, BCLT effectiveness did not seem to be different in the two groups and was partially additive with gefitinib in the NHT group and additive/synergistic in the control group. Cultures (8/22) were negative for the expression of the PTEN gene and we observed no differences in the two groups. Thus the different IC50 values observed for gefitinib and the partial additivity in the combination treatment with gefitinib and BCLT is influenced by EGFR/Her2 ratio because it was shown that EGFR inhibition was lower when Her2 is overexpressed. Taken together, our results indicate that anti-EGFR targeted therapies and a possible combination therapy involving gefitinib and BCLT should be performed early in naive patients when Her2 is not overexpressed and before the anti-androgenic hormone therapy induces such an undesirable effect. PMID- 17914593 TI - Expression of phospholipase D2 in human colorectal carcinoma. AB - Phospholipase D (PLD) catalyzes the hydrolysis of phosphatidylcholine (PC) to generate phosphatidic acid (PA) and choline. PA acts as a second messenger in cell proliferation; therefore PLD is believed to play an important role in carcinogenesis. PLD activity has been reported to be elevated in human breast, gastric, renal cell and colorectal carcinomas, compared with adjacent non neoplastic tissues. The activity of PLD was also correlated with nuclear grade in breast cancer, tumor size in gastric carcinoma, and nodal involvement and deeper invasion in colorectal carcinoma. However, the number of cases in each study was small. The aim of this study was to investigate the expression level of PLD2 and its association with clinicopathological features in human colorectal carcinoma. Ninety-seven colorectal carcinomas were obtained from surgery. Expression level of PLD2 was assessed by real-time PCR. The prognostic relevance of PLD2 expression level in patients with colorectal carcinoma was also analyzed by the survival analysis of mortality follow-up data covering the period 2000-2004. PLD expression level was varied from tumor to tumor. Expression level of PLD was significantly correlated with tumor size (P<0.05); it was independent of lymph node metastasis, extent of invasion, pathological classification, distant metastasis and Dukes' stage. PLD expression level was also significantly correlated with survival of patients with colorectal carcinoma (P<0.05). These findings suggested that PLD2 plays an important role in progression of colorectal carcinoma and that PLD2 could be a target for therapy in colorectal carcinoma. PMID- 17914594 TI - Local control in 118 consecutive high-risk breast cancer patients treated with breast-conserving therapy. AB - Although negative surgical margins are an important prognostic factor in the breast-conserving treatment of breast cancer, the required width of these margins is still under debate. To define the risk of in-breast recurrence in subgroups of patients with a local high-risk situation, local control was evaluated in all patients with close or positive margins treated at one institution between 1995 and 2000. A total of 118 patients (67% T1, 30% T2, 52% N0) were identified as having had positive or close margins (< or =4 mm) at the time of initial surgery. Of these, 65% had no tumor cells at the initial margin, 35% had a positive or questionable margin. Re-excisions were performed in 42%. The median (range) whole breast radiotherapy dose, tumor bed boost dose and total dose were 50 (46.8-56) Gy, 15 (8-20) Gy and 65.8 (54-71) Gy, respectively. Thirty-six percent received adjuvant chemotherapy. Local (in-breast) control was calculated by the Kaplan Meier method and compared between subgroups. The 5-year local control for the whole group was 94%. The rates for selected subgroups were: <56 years 89.4% vs. >56 years 98.1% (p=0.073, univariate analysis); pT1 95.9% vs. pT2 88.6% (not significant, n.s.); pN0 96.6% vs. pN+ 90.8% (n.s.); initial margin free of tumor cells 95.5% vs. initial margin involved or questionable 90.7% (n.s.), no re excision 96.7% vs. one or more re-excisions 90.6% (n.s.); adjuvant chemotherapy 81.7% vs. no adjuvant chemotherapy 100% (p=0.007). We conclude that among patients with close or positive margins, older patients achieved high local control rates with a median tumorbed boost to 66 Gy. Younger patients and patients who received adjuvant chemotherapy (due to the presence of histopathological risk factors) were at increased risk of in-breast recurrence and should be considered for intensified local treatment. PMID- 17914595 TI - A review of ductal stenting in hypoplastic left heart syndrome: bridge to transplantation and hybrid stage I palliation. AB - There is increasing interest in applying ductal stenting technology to high-risk patients with hypoplastic left heart syndrome (HLHS). In this review, we present the complete history and a comprehensive up-to-date analysis of all available data on the use of ductal stenting as part of various hybrid strategies for the combined medical and surgical management of HLHS. PMID- 17914596 TI - A surgical algorithm using open rhinoplasty for correction of traumatic twisted nose. PMID- 17914597 TI - Lower lid blepharoplasty: the use of Lockwood's ligament for orbicularis oculi suspension and orbital fat preservation-a new technique. AB - A new technique of lower lid blepharoplasty is presented. The technique consists of suspending the skin-muscle flap of the lower lid on the suspensory Lockwood's ligament in all the orbit length, with preservation of the orbital fat behind the created anatomic layer. The anatomic base of the technique is analyzed by a study of the literature, by personal observations of the changes in the anatomic elements of the orbit in a large number of operations the authors performed for patients with endocrinic exophthalmos using Olivari's method (transpalpepral orbital decompression by removal of the orbital fat), and by fresh cadaver dissections. The technique is described in detail, and the results are presented. After 8 years of follow-up evaluation, it can be confidently said that the proposed technique can solve almost all the problems associated with lower lid laxity and can be applied for the majority of patients. PMID- 17914598 TI - Surgical disconnection of cortical venous reflux as a treatment for Borden type II dural arteriovenous fistulae. AB - OBJECTIVE: The presence of cortical venous reflux is recognized as an indicator of increased risk of intracranial hemorrhage and neurological deficits in cranial dural arteriovenous fistulas. Its disconnection is well accepted as a treatment for fistulas with direct cortical reflux (Borden type III), but the role of disconnection of the cortical venous reflux in the management of fistulas that involve the venous sinus and cortical venous reflux (Borden type II) is still a matter of debate. We analyze the experience of the Toronto Brain Vascular Malformation Study Group in the management of these lesions by simple cortical venous reflux disconnection and its impact in the future risk of bleeding. METHODS: From June 1984 to August 2004, 347 patients with dural arteriovenous fistulas, either cranial or spinal, were evaluated by the group. Fifty-three patients had a Borden type II dural arteriovenous fistulas. Twenty-five patients were submitted to simple surgical disconnection of the dural arteriovenous fistulas, two were lost for follow-up. There were 15 females and 8 males, with mean age at diagnosis of 53.9 years. Follow-up time was 112.6 patient-years, from 2 months to 11 years, mean 4.9 years. Endovascular treatment was attempted in all patients, but no disconnection was possible. Twelve patients had their fistulas completely occluded by endovascular means, but are not analyzed here. There were four complications from the 93 endovascular procedures, and 3 from the 27 surgical procedures. Two patients required a repeated surgical procedure. No episode of intracranial hemorrhage or worsening neurological deficit was seen after disconnection of the cortical venous reflux in 4.9 years of follow-up. CONCLUSION: Simple surgical disconnection of the cortical venous reflux maybe an option in the management of patients with Borden type II dural arteriovenous fistulas. This procedure is a much smaller surgical undertaking and is associated with fewer complications than attempts to resect or pack the whole fistula, especially if located in the skull base. PMID- 17914599 TI - Pituitary adenoma and concomitant Rathke's cleft cyst. AB - Although pituitary adenomas and Rathke's cleft cysts have a shared ancestry, they rarely occur simultaneously. Only 32 reports involving a pituitary adenoma and a concomitant Rathke's cleft cyst were identified upon review of the literature. Most initial presenting complaints include hormonal symptoms, visual disturbances, and headache. Next to growth hormone, Prolactin was the most commonly hypersecreted pituitary hormone. Rathke's cleft cysts show variable position, size, and signal intensity on magnetic resonance imaging (MRI). Here, we report a patient with a growth hormone- secreting pituitary adenoma associated with a Rathke's cleft cyst. The mass contained two different signal intensities on MRI. The lesion was successfully removed assisted by intraoperative MRI, when the presence of both lesions was confirmed. When a non-enhancing cyst-like structure is demonstrated on imaging in a patient with a pituitary adenoma, the possibility of a coexisting Rathke's cleft cyst should be considered. PMID- 17914600 TI - A concise, asymmetric synthesis of (2R,3R)-3-hydroxyaspartic acid. AB - 3-Hydroxyaspartic acid and its derivatives are found both in the free form and as peptide constituents in various microorganisms and fungi. Considering the biological importance of this amino acid and its potential utility as a multifunctional building block in organic syntheses, we have developed a short step, asymmetric synthetic route to a strategically protected 3-hydroxyaspartic acid derivative in enantiopure form. The key steps in the synthesis involve, Sharpless asymmetric aminohydroxylation of commercially available trans-ethyl cinnamate, and, utilization of the phenyl group as a masked carboxylic acid synthon towards construction of the complete structural framework of the title compound. PMID- 17914601 TI - Design and synthesis of a novel anthracene-based fluorescent probe through the application of the Suzuki-Miyaura cross-coupling reaction. AB - We report on a simple synthetic route to a novel anthracene-based bis-armed amino acid derivative as a useful fluorescent probe. Various photophysical studies of this amino acid derivative are also described. Here, Suzuki-Miyaura cross coupling reaction has been used as a key step for carbon-carbon bond formation. PMID- 17914602 TI - Kinetics of the photosensitized oxidation of chymotrypsin in different media. AB - The kinetic aspects of the Perinaphthenone-sensitized photooxidation (singlet molecular oxygen [O2 ((1)Delta(g))]-mediated) of alpha-chymotrypsin (alpha-Chymo) have been studied at pH 8 and pH 11 as well in reverse micelles (RMs) of sodium 1, 4 bis (2-ethylhexyl) sulfosuccinate (AOT) in n-heptane. The rate constant values for both overall (k(t)) and chemical (k(r)) quenching of O2 ((1)Delta(g)) by alpha-Chymo in homogeneous media were higher at pH = 11 than at pH = 8, indicating that the OH-ionized tyrosine (Tyr) residues, clearly dominate the quenching process. Besides, the rate constants in water were higher than those determined in RMs, demonstrating that the organized medium protects the protein against photooxidation, probably due to a diminution in both, the accessibility towards oxidizable amino acid residues and the polarity inside the aggregate, as compared to water. The protection effect of alpha-Chymo against the attack by the oxidative species O2 ((1)Delta(g)) in RMs of AOT seems to be due to the increase of protein stability by the encapsulation within the micellar structure. The effect of both, surfactant concentration and variation of the ratio ([H2O]/[AOT]) = W on the reactive rate constant was also investigated. The process does not depend significantly on micelles concentration while the k(r) values increase as W increases. Furthermore, at W = 30, the highest W studied, k(r) tends to the value obtained in aqueous medium. PMID- 17914604 TI - Hepatitis C virus entry into host cells. AB - The recent development of functional models to analyze the early steps of the hepatitis C virus (HCV) life cycle has highlighted that HCV entry is a slow and complex multistep process involving the presence of several entry factors. Initial host cell attachment may involve glycosaminoglycans and the low-density lipoprotein receptor, after which the particle appears to interact sequentially with three entry factors: the scavenger receptor class B type I, the tetraspanin CD81 and the tight-junction protein claudin-1. Several serum components may also modulate HCV entry, while the recently discovered CD81 partner EWI-2wint can block the interaction of the viral particle with CD81, potentially preventing infection in the cell types in which it is expressed. After binding to the host cell, the HCV particle is internalized by clathrin-mediated endocytosis, with fusion likely occurring in early endosomes. This review summarizes our current knowledge on HCV entry. PMID- 17914603 TI - Production of methanethiol and volatile sulfur compounds by the archaeon "Ferroplasma acidarmanus". AB - Acidophiles are typically isolated from sulfate-rich ecological niches yet the role of sulfur metabolism in their growth and survival is poorly defined. Studies of heterotrophically grown "Ferroplasma acidarmanus" showed that its growth requires a minimum of 100 mM of a sulfate-containing salt. Headspace gas analyses by GC/MS determined that the volatile sulfur compound emitted by active "F. acidarmanus" cultures is methanethiol. In "F. acidarmanus" cultures grown either heterotrophically or chemolithotrophically, methanethiol was produced constitutively. Radiotracer studies with (35)S-labeled methionine, cysteine, and sulfate showed that all three were used in methanethiol production. Additionally, (3)H-labeled methionine was incorporated into methanethiol and was probably used as a methyl-group donor. Methanethiol production in whole cell lysates supplied with SO (3) (2-) indicated that NADPH-dependant sulfite reductase and methyltransferase activities were present. Cell lysates also contained enzymatic activity for methionine-gamma-lyase that cleaved the side chain of either methionine to form methanethiol or cysteine to produce H(2)S. Since methanethiol was detected from the degradation of cysteine, it is likely that sulfide was methylated by a thiol methyltransferase. Collectively, these data demonstrate that "F. acidarmanus" produces methanethiol through the metabolism of methionine, cysteine, or sulfate. This is the first report of a methanethiol-producing acidophile, thus identifying a new contributor to the global sulfur cycle. PMID- 17914605 TI - Periorbital necrobiosis lipoidica diabeticorum: case report. AB - Necrobiosis lipoidica diabeticorum (NLD) is a relatively common manifestation of diabetes mellitus that is thought to be related to immune-complex vasculitis. The legs are the most common site for NLD, but other locations, such as the forearms or hands, have been reported. We describe a case of periorbital NLD that was diagnosed on the basis of histopathology. PMID- 17914606 TI - Heterologous expression of a gene encoding a thermostable beta-galactosidase from Alicyclobacillus acidocaldarius. AB - A genomic DNA library screen yielded the nucleotide sequence of a 12 kb fragment containing a gene (2067 bp) coding a thermostable beta-galactosidase from Alicyclobacillus acidocaldarius ATCC 27009. The beta-galactosidase gene was expressed in Pichia pastoris, and up to 90 mg recombinant beta-galactosidase/l accumulated in shake flask cultures. Using o-nitrophenyl-beta-D: galactopyranoside as a substrate, the optimum pH and temperature of the purified recombinant beta-galactosidase were 5.8-6.0 and 70 degrees C, respectively. The enzyme retained 90% of its activity when heated at 70 degrees C for 30 min. Approximately 48% of lactose in milk was hydrolyzed following treatment with the recombinant enzyme over 60 min at 65 degrees C. PMID- 17914607 TI - A novel biotransformation of 2-formyl-6-naphthoic acid to 2,6-naphthalene dicarboxylic acid by Pseudomonas sp. for the purification of crude 2,6 naphthalene dicarboxylic acid. AB - Crude 2,6-naphthalene dicarboxylic acid was purified by Pseudomonas sp. HN-72 which biotransformed the major impurity of 2-formyl-6-naphthoic acid into 2,6 naphthalene dicarboxylic acid. The biotransformation yield reached 100% when the reaction was performed at 40 degrees C for 1 h, in 200 ml KH(2)PO(4)/KOH buffer (50 mM, pH 8.0), with 0.2% (w/v) crude 2,6-naphthalene dicarboxylic acid and 2.5 mg dry cell wt/ml. PMID- 17914608 TI - Release of short chain fatty acids from cream lipids by commercial lipases and esterases. AB - Lipases and esterases are frequently used in dairy production processes to enhance the buttery flavour of the end product. Short chain fatty acids, and especially butanoic acid, play a key role in this and different enzymes with specificity towards short chain fatty acids are commercially available as potent flavouring tools. We have compared six lipases/esterases associated with buttery flavour production. Although specificity to short chain fatty acids was ascribed to each enzyme, clear differences in free fatty acid profiles were found when these enzymes were applied on cream. Candida cylindraceae lipase was the most useful enzyme for buttery flavour production in cream with the highest yield of free fatty acids (57 g oleic acid 100 g(-1) fat), no release of long chain fatty acids and specificity towards butanoic acid. PMID- 17914609 TI - The kinetics of water loss from zinc phosphate and zinc polycarboxylate dental cements. AB - The water desorption behaviour of three different zinc oxide dental cements (two polycarboxylates, one phosphate) has been studied in detail. Disc-shaped specimens of each material were prepared and allowed to lose water by being subjected to a low humidity desiccating atmosphere over concentrated sulfuric acid. In all three cements, water loss was found to follow Fick's second law for at least 6 h (until M(t)/M(infinity) values were around 0.5), with diffusion coefficients ranging from 6.03 x 10(-8 )cm(2 )s(-1) (for the zinc phosphate) to 2.056 x 10(-7 )cm(2 )s(-1) (for one of the zinc polycarboxylates, Poly F Plus). Equilibration times for desorption were of the order of 8 weeks, and equilibrium water losses ranged from 7.1% for zinc phosphate to 16.9% and 17.4% for the two zinc polycarboxylates. PMID- 17914610 TI - Influence of spoliation in poly(2-hydroxy ethyl methacrylate) soft contact lens on its free volume and optical transparency. AB - The calcification in poly(2-hydroxy ethyl methacrylate) contact lens was investigated using positron annihilation spectroscopy (PLS). The two poly(2 hydroxy ethyl methacrylate) (PHEMA) lenses of different companies were calcified employing a simple mechanism of calcification in abiotic aqueous solutions. The calcium deposit was analyzed using energy dispersive X-ray spectroscopy (EDS). Calcified lenses showed decrease in ortho-positronium (o-Ps) lifetime and free volume hole size of the lens material suggesting diffusion of Ca2+ into these cavities. The change in optical property viz. refractive index of these calcified lenses were measured and correlated with positron results. To find a better correlation, a series of worn spoilt PHEMA lenses of the same power with mainly calcium deposits, were similarly characterized using PLS and refractive index. These results correlate well with the free volume of the material. For hydrophilic lenses this correlation is reported for the first time. PMID- 17914611 TI - In vivo and in vitro response to electrochemically anodized Ti-6Al-4V alloy. AB - Tissues' reactions to metals depend on a variety of properties of the metal, most notably surface structure. Anodizing has been shown to alter the surface properties of metal, thus eliciting a change in the biocompatibility of the metal. In order to evaluate the biocompatibility of unoxidized titanium alloy (Ti 6Al-4V) and anodized titanium alloy samples, the samples were implanted in murine abdominal subcutaneous tissues, and maintained for 2 and 4 weeks. The reaction of the abdominal subcutaneous connective tissues to the samples was then assessed. Fibrous connective tissue capsules were observed around the vicinity of the sample, and these capsules were shown to harbor fibroblasts, fibrocytes, and other cells, including neutrophils, macrophages, and giant multinucleated cells. The average thickness of the fibrous capsules observed around the anodized alloy samples was less than that of the capsules seen around samples of the unoxidized titanium alloy. Blood was obtained from the tails of the experimental mice, and blood cell analyses were conducted in order to assess the levels of leukocytes, red blood cells, and thrombocytes. The blood analysis results of the unoxidized control group and treatment group were all within normal ranges. In addition, the biocompatibility of the titanium alloy samples was evaluated using cell culture techniques. The numbers of MG-63 cells cultured on oxidized samples tended to be greater than those in the controls; however, these increases were not statistically significant. The alkaline phosphatase activity of the sample oxidized at 310 V evidenced significantly higher activity than was observed in the control group. These results indicate that the anodized Ti-6Al-4V alloy will be of considerable utility in biomedical applications. PMID- 17914612 TI - Thin films of SiO2 and hydroxyapatite on titanium deposited by spray pyrolysis. AB - Wet spray pyrolysis of fine, well-dispersed a SiO2 sol was used for the deposition of thin films of silicon dioxide. The sol was obtained by hydrothermal precipitation of silicon acid from a solution at pH = 10. The morphology, roughness, phase composition, chemical homogeneity and the mechanism of the films were investigated by SEM, EDS and IR spectroscopy. The obtained results show a complete covering of the titanium substrate with SiO2 after 3 h of deposition. It was observed that the film thickness increased from 3 to 19 microm, the roughness of the film decreased from 12 to 3 microm, while the morphology of the deposit changed considerably. A hydroxyapatite film was prepared on the so-obtained SiO2 thin film by spray pyrolysis deposition and its morphology and phase composition were investigated. PMID- 17914613 TI - Laser surface processing of Ti6Al4V in gaseous nitrogen: corrosion performance in physiological solution. AB - Laser surface processing was carried out in gaseous nitrogen atmosphere at ambient temperature. The laser scan speed was varied (50-150 cm/min) at constant power of 1500 watts and resulting changes such as microstructural evolution, hardness, and electrochemical response of modified surface in Ringer's physiological solution at varying pH were studied. Increase in laser scanning speed was found to reduce the thickness of alloyed zone from 258 to 87 microm. The microstructure of laser-modified surface contains dendrites grown perpendicular to the laser traverse direction, beneath which basket weave structure of acicular alpha (martensite) was prevalent. Hardness at the top surface of laser-processed at 50 cm/min was approximately 1137 kg/mm2 that reduced with increase in the laser scan speed (577 kg/mm2 at 150 cm/min). Laser surface processing shifted the corrosion potential of Ti6Al4V towards noble side as compared to untreated alloy; the maximum shift by approximately 494 mV was recorded in pH approximately 9 solution. Passivation after laser surface modification was improved as currents were at least 1/3 of the untreated Ti6Al4V in passive region. While the pitting potential of untreated material was found to increase from 1.84 V for 4.0 pH to >2.5 V for 9.0 pH, the pitting potential after laser treatment was observed to drop from maximum of 74% for 4.0 pH (at 100 cm/min) to maximum of 42% for 9.0 pH (at 150 cm/min). PMID- 17914614 TI - In vitro heat generation by ferrimagnetic maghemite microspheres for hyperthermic treatment of cancer under an alternating magnetic field. AB - Ferrimagnetic materials can be expected to be useful as thermo seeds for hyperthermic treatment of cancer, especially where the cancer is located in deep parts of body, as they can generate heat by magnetic hysteretic loss when they are placed in an alternating magnetic field. Recently, it was reported that ferrimagnetic maghemite (gamma-Fe2O3) microspheres 20-30 microm in diameter prepared in aqueous solution can show excellent heat generating ability. However, these microspheres have many cracks on their surfaces. In this study, the preparation conditions for the microspheres was further optimized in order to obtain crack-free ferrimagnetic microspheres, and the in vitro heat generation of the obtained microspheres was measured in an agar phantom under an alternating magnetic field. Crack-free gamma-Fe2O3 microspheres 20-30 microm in diameter were obtained successfully. Their saturation magnetization and coercive force were 68 emu g(-1) and 198 Oe, respectively. Their heat generation under an alternating magnetic field of 300 Oe at 100 kHz was estimated to be 42 W g(-1). The microspheres showed in vitro heat generation when they were dispersed in an agar phantom and placed under an alternating magnetic field. It is believed that these microspheres may be useful for the in situ hyperthermic treatment of cancer. PMID- 17914615 TI - Synthesis and hydrothermal treatment of nanostructured hydroxyapatite of controllable sizes. AB - Nanoparticulate systems have been studied for targeted and controlled release of therapeutic agents; and size is one of the major determinants of their in vivo clearance kinetics by the MPS macrophages. As such, it is important to control the size of hydroxyapatite nanoparticles during synthesis. The results show that the size of hydroxyapatite nanoparticles, synthesized through chemical precipitation, increases with increasing synthesis time. Particle sizes were also observed to increase in a linear correlation with temperature. Crystallinity and carbonate-substitution of the nanoparticles also increased with temperature. Hydrothermal, performed as a post-synthesis treatment, improves particle morphology, giving particles with regular surface contours, well-defined sizes and lower particle agglomeration. By controlling synthesis temperature and time, hydroxyapatite nanoparticles with well-defined sizes and morphology can be obtained. PMID- 17914616 TI - Improvement in crystallinity of apatite coating on titanium with the insertion of CaF2 buffer layer. AB - In the apatite coatings on Ti the heat treatment process is necessary to crystallize the apatite structure for improved chemical stability and biological properties. However, the heat treatment normally degrades the mechanical strength of the coating layer associated with thermally induced stress. In this study, we aimed to improve the crystallization of apatite coating by using calcium fluoride (CaF2) as a buffer layer. The insertion of a thin layer of CaF2 (0.2-1 microm) between apatite and Ti significantly improved the crystallization behavior of apatite. Moreover, this crystallization was more enhanced as the thickness of CaF2 was increased. When a 1 microm-thick CaF2 was inserted, the crystallization of apatite initiated at a temperature as low as 320 degrees C, being a dramatic improvement in the crystallization when considering the crystallization initiation temperature of a bare apatite coating on Ti was approximately 450 degrees C. As a result of this crystallization enhancement, the dissolution behavior of CaF2-inserted apatite coatings was more stable than that of the bare apatite coating, showing much reduced initial-burst effect. Preliminary cellular assay showed the CaF2-inserted apatite coating provided a substrate for cells to spread and grow favorably, as being similar to the bare apatite coating. This novel way of apatite coating on Ti using CaF2 buffer layer may be useful in the coating systems particularly requiring low temperature processing and increased crystallinity with high chemical stability. PMID- 17914617 TI - Sol-gel preparation and high-energy XRD study of (CaO)x(TiO2) 0.5-x(P2O5)0.5 glasses (x = 0 and 0.25). AB - Glasses from the CaO-TiO2-P2O5 system have potential use in biomedical applications. Here a method for the sol-gel synthesis of the ternary glass (CaO)0.25(TiO2)0.25(P2O5)0.5 has been developed. The structures of the dried gel and heat-treated glass were studied using high-energy X-ray diffraction. The structure of the binary (TiO2)0.5(P2O5)0.5 sol-gel was studied for comparison. The results reveal that the heat-treated (CaO)0.25(TiO2)0.25(P2O5)0.5 glass has a structure based on chains and rings of PO4 tetrahedra, held together by a combination of electrostatic interaction with Ca2+ ions and by corner-sharing oxygen atoms with TiO6 octahedra. In contrast, the (TiO2)0.5(P2O5)0.5 glass has a structure based on isolated P2O7 units linked together by corner-sharing with TiO6 groups. The results suggest that both the dried gels possess open porous structures. For the (CaO)0.25(TiO2)0.25(P2O5)0.5 sample there is a significant increase in Ca-O coordination number with heat treatment. PMID- 17914619 TI - Relationship between apatite-forming ability and mechanical properties of bioactive PMMA-based bone cement modified with calcium salts and alkoxysilane. AB - Polymethylmethacrylate (PMMA)-based bone cement is used for the fixation of artificial joints in orthopaedics. However, the fixation is liable to loosen in the body, because the cement does not bond to living bone. So-called bioactive ceramics bond directly to living bone through the apatite layer formed on their surfaces in the body. We previously revealed that modification using gamma methacryloxypropyltrimethoxysilane (MPS) and water-soluble calcium salts such as calcium acetate and calcium hydroxide was effective for providing the PMMA-based bone cement with apatite-forming ability in a simulated body fluid (SBF, Kokubo solution) that closely reproduces the body environment. However, the effect of the chemical reaction forming the apatite on the mechanical properties of the cements has not been clarified. The present work aimed to investigate this issue from the viewpoint of the interface structure between the apatite and the cement. The surface of the cement modified with calcium acetate and MPS was fully covered with newly formed apatite after soaking in Kokubo solution within 7 days, while half of the surface area of the cement modified with calcium hydroxide and MPS was covered with the apatite. The bending strength of the modified cements decreased after soaking in Kokubo solution. Porous structure was observed in the region about 50-100 microm in depth from the top surface because of release of the Ca2+ ions by both modified cements after soaking in Kokubo solution. The decrease in bending strength of the modified cements could be attributed to the formation of the pores. In addition, the pores on the top surfaces of the cements were filled with the newly formed apatite. The apatite formation would be effective not only for bioactivity but also for decreasing the reduction of mechanical strength. PMID- 17914618 TI - Controlled release of vancomycin from cross-linked gelatine. AB - This paper explores the possibility of using biodegradable cross-linked gelatines as antibiotic devices for a long-term elution (80 days). Capillary electrophoresis (CE) has been utilized to evaluate the mass percentage of vancomycin and gelatine contemporary released from differently cross-linked vancomycin loaded gelatine samples in an elution time ranging from 24 to 1920 h. While the solubilization kinetic of gelatine samples differently cross-linked can be very close described by the simplified Higuchi model, the vancomycin release kinetic is contemporary governed by both the Fickian diffusion process trough the gelatine matrix network and the dissolution process of the matrix due to its degradation. Comparing the antibiotic eluting kinetics from gelatine at diverse cross-linking degree we observed that the degradation of the proteic matrix appears to have a minor influence in the drug release control. Vancomycin released from all the gelatine partially cross-linked samples results active against Staphylococcus aureus and Streptococcus faecalis which represent the most pathogens commonly isolated in orthopaedic infections. Vancomycin overcomes the minimum inhibitory concentration for both the bacteria in the whole range of elution time. Cross-linked gelatine devices appear to represent a useful biodegradable delivery system for local anti-infective therapy in arthoplasty. PMID- 17914620 TI - Biological responses of novel high-toughness double network hydrogels in muscle and the subcutaneous tissues. AB - The study evaluated biological reaction of four types of novel double network gels in muscle and subcutaneous tissues, using implantation tests according to the international guideline. The implantation tests demonstrated that, although poly (2-acrylamide-2-metyl-propane sulfonic acid)/poly (N,N'-dimetyl acrylamide) (PAMPS/PDMAAm) gel induced a mild inflammation at 1 week, the degree of the inflammation significantly decreased into the same degree as that of the negative control at 4 and 6 weeks. This gel has a potential to be applied as artificial cartilage. In addition, Cellulose/Gelatin gel showed the same degree of inflammation as that of the negative control at 1 week, and then, showed a gradually absorbable property at 4 and 6 weeks. This gel has a potential to be applied as an absorbable implant. The PAMPS/polyacrylamide and Cellulose/PDMAAm gels induced a significant inflammation at each week. These DN gels are difficult to be applied as clinical implants in the current situation. PMID- 17914621 TI - Creatinine biomaterial thin films grown by laser techniques. AB - Creatinine thin films were synthesised by matrix assisted pulsed laser deposition (PLD) techniques for enzyme-based biosensor applications. An UV KrF* (lambda=248 nm, tau approximately 10 ns) excimer laser source was used for the irradiation of the targets at incident fluence values in the 0.3-0.5 J/cm2 range. For the matrix assisted PLD the targets consisted on a frozen composite obtained by dissolving the biomaterials in distilled water. The surface morphology, chemical composition and structure of the obtained biomaterial thin films were investigated by scanning electron microscopy, Fourier transform infrared spectroscopy, and electron dispersive X-ray spectroscopy as a function of the target preparation procedure and incident laser fluence. PMID- 17914622 TI - Engineering of poly(epsilon-caprolactone) microcarriers to modulate protein encapsulation capability and release kinetic. AB - Drug delivery applications using biodegradable polymeric microspheres are becoming an important means of delivering therapeutic agents. The aim of this work was to modulate the microporosity of poly(epsilon-caprolactone) (PCL) microcarriers to control protein loading capability and release profile. PCL microparticles loaded with BSA (bovine serum albumin) have been de novo synthesized with double emulsion solvent evaporation technique transferred and adapted for different polymer concentrations (1.7 and 3% w/v) and stabilizer present in the inner aqueous phase (0.05, 0.5 and 1% w/v). SEM (scanning electron microscope) and CLSM (confocal laser scanning microscope) analysis map the drug distribution in homogeneously distributed cavities inside the microspheres with dimensions that can be modulated by varying double emulsion process parameters. The inner structure of BSA-loaded microspheres is greatly affected by the surfactant concentration in the internal aqueous phase, while a slight influence of polymer concentration in the oil phase was observed. The surfactant concentration mainly determines microspheres morphology, as well as drug release kinetics, as confirmed by our in-vitro BSA release study. Moreover, the entrapped protein remained unaltered during the protein encapsulation process, retaining its bio-activity and structure, as shown through a dedicated gel chromatographic analytical method. PMID- 17914623 TI - The role of aluminium and silicon in the setting chemistry of glass ionomer cements. AB - A model of the setting chemistry of glass-ionomer cements (GICs) is proposed based on 27Al and 29Si solid state nuclear magnetic resonance spectroscopy data on three GICs. All the precursor glasses are found to contain three aluminium species viz.: four, five and six-coordinate aluminium environments as well as four-bridging silicate tetrahedra. Upon cement formation, Al3+ ions in the glass are leached out from the surface layer of the glass. On entering the cement matrix, these ions adopt six-coordination and crosslink the polymer chains as part of the setting reaction. The remaining four-coordinate aluminium is distributed between two species: one in the inert core of the glass particles; and a second, less concentrated, in the surface layer of the glass particles, modified by the curing reactions. There is some evidence for residual five and six coordinate aluminium species in the final cement in some of the systems. In the case of the silicate tetrahedra, the curing reactions result in a decrease in the number of aluminium atoms in the second coordination sphere, with a subsequent recondensation of silicate network of the glass. PMID- 17914624 TI - Evaluation of the release rate of bioactive recombinant human epidermal growth factor from crosslinking collagen sponges. AB - The purpose of this study was to prepare recombinant human epidermal growth factor (rhEGF) collagen sponges for topical applications and investigate the effects of different types of crosslinked collagen sponges as platforms for the controlled release of rhEGF. The microstructure and the drug release rates of collagen sponges were modified through treatment with different types (glutaraldehyde (GTA), genipin and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)), different concentrations of crosslinking agents and various preparation conditions. A controlled release profile was observed for the crosslinked collagen sponges as compared to the non-crosslinked ones. The results indicated that the GTA crosslinked sponges have the most potent controlling effect. As the amount of GTA increased, a greater rigidity of the collagen sponge structure combined with a lower hydrophilicity was observed, leading to a decreased drug release rate and an increased water uptake. This study also demonstrated that a good correlation was obtained for in vitro release rates of rhEGF using the power model. The crosslinked rhEGF collagen sponges showed a successful delivery of rhEGF in bioactive form to stimulate cell proliferation. PMID- 17914625 TI - On the biocompatibility of a novel Ti-based amorphous composite: structural characterization and in-vitro osteoblasts response. AB - Titanium and its alloys are frequently used as dental and orthopaedic implants due to their high mechanical strength, low elastic modulus and biocompatibility. However, as these materials have a poor wear resistance, tribo-chemical reactions during use produce debris accumulation, resulting in adverse cellular responses. In that sense, amorphous based materials are potential candidates, considering their hardness and crack growth resistance. This paper reports on the structural characterization of the as-quenched Ti45Zr38Ni17 alloy. This system displays a duplex structure never mentioned before with a low dispersion of nanometric beta phase particles in an amorphous matrix. Moreover, in order to explore the biocompatibility of such composite, primary osteoblasts cultures are used to analyse cell behaviour around and upon the metallic surface. Osteoblasts attach and proliferate on the material as demonstrated by scanning electron microscopy. Cell proliferation and bone nodule formation are also observed in cultures with Ti45Zr38Ni17 particles by phase contrast microscope. In addition, transmission electron microscopy reveals ultrastructural features very close to those observed in vivo during intramembranous ossification with active osteoblasts surrounded by an extracellular matrix and a mineralized one. In conclusion, these results demonstrate that osteoblasts, cultured in presence of Ti45Zr38Ni17 alloy, proliferate, differentiate and synthesize bone matrix. PMID- 17914626 TI - Effect of treated filler loading on the photopolymerization inhibition and shrinkage of a dimethacrylate matrix. AB - This study shows how treated filler loading influences the photopolymerization of a dimethacrylate comonomer mixture, regarding, in particular, shrinkage and inhibition under atmospheric oxygen, present in oral environment. Bis-GMA/TEGDMA (75/25 wt.%) resins were loaded with hybrid filler (Ba aluminosilicate glass and pyrogenic silica), treated with gamma-methacryloxy(propyl)trimethoxysilane, at 0 50 wt.% and light cured over a total of 30 s (45 mW/cm2). Degree of double-bond conversion (DC), obtained using FTIR, decreased with filler content. 1H MAS spectra (293-340 K) and STRAFI images (293 K) were obtained as a function of irradiation time and filler concentration. 1H signals of unreacted methacrylate groups were more intense for higher loaded resins and resonances from CH2SiO2(OH) (T2) and -CH2SiO3- (T3) units, also observed by 29Si NMR, were resolved and suggest the presence of T2-resin bonds. 1D images show a reduction on polymerization contraction and reaction inhibition at the composite resin surface with filler loading. 2D resin images present a highly mobile surface layer, hence with lower DC. PMID- 17914627 TI - Antibacterial activity of ZnO powder with crystallographic orientation. AB - ZnO powder with crystallographic orientation was prepared from the mixed aqueous solution of zinc chloride, tri-ethanol amine and thio-urea. From X-ray diffraction measurement, as-prepared powder was found to have the orientation along a-b axes of hexagonal structure, and a needle-like shape with the aspect ratio of 5 was observed by scanning electron microscope, indicating that as prepared powder had crystallographic orientation. In the tests of antibacterial activity by colony count method, ZnO powders with and without crystallographic orientation were used in present work. Survival ratio of bacteria decreased with increasing powder concentration, i.e., increase in antibacterial activity. The antibacterial activity in ZnO powder with crystallographic orientation was weaker than that in commercial ZnO powder without orientation at same powder concentration. Regarding specific surface area of the powders used in antibacterial tests, however, antibacterial activity in powder with orientation was found to be similar to that without orientation; that is, the crystallographic orientation of ZnO did not affect antibacterial activity. The activity toward Staphylococcus aureus was stronger than that toward Escherichia coli, irrespective of the kind of powders. PMID- 17914629 TI - Experimental validation of a new approach for the development of mechano compatible composite scaffolds for vascular tissue engineering. AB - The clinical need for the design of small-diameter vascular substitutes with high patency rates has never been so urgent as nowadays. Mechano-compatibility is widely known as one of the main key parameter for the design and the development of highly-patent vascular substitutes independently of their nature, i.e., arterial prostheses, arterial grafts or tissue-engineered blood-vessel. In this work, we attempt to target mechano-compatibility of cylindrical scaffolds for vascular tissue engineering by a computational model based on the composite theory associated with finite element and genetic algorithm. Then, cylindrical composite scaffolds were fabricated from gelatine (matrix) and silk (reinforcement) to experimentally validate theoretical results obtained by the implemented computational model. Finally, the compliance of the scaffolds was measured by an in-house developed specific device. Results show that the computational predictions from numerical simulation are in good agreement with the measurements obtained form the experimental tests. Therefore, the proposed computational model represents a valid tool to assist biomaterial scientists during the design of composite scaffolds, and especially in targeting their mechanical properties. PMID- 17914628 TI - Preparation and blood coagulation evaluation of chitosan microspheres. AB - Cross-linked chitosan microspheres (40-100 microm) with smooth surface were prepared by the methods of emulsification and ethanol coagulant. FTIR results showed that the cross-linking reaction occurred on the amino groups of chitosan molecules. The swelling characteristic of chitosan microspheres was influenced by the environment pH, being generally greater at low rather than higher pH values. The coagulation properties of chitosan microspheres were evaluated by dynamic blood clotting, platelet adhesion and activation, erythrocyte adhesion, hemolysis, and protein absorption assays. Chitosan microspheres can shorten the clotting time and induce the adhesion and activation of platelets. But the shortening of clotting time by chitosan microspheres may be related to not only platelet aggregation, but also erythrocyte aggregation. Take together, chitosan microspheres may be potential use as thrombospheres. PMID- 17914630 TI - Intraosseous pressure and strain generated potential of cylindrical bone samples in the drained uniaxial condition for various loading rates. AB - Cortical bone is a composite material consisting of a porous elastic solid and viscous fluid. It is well known that the intraosseous fluid circulates as a result of a bone fluid pressure gradient in the porous space of the cortical bone. When a time-dependent mechanical load is applied to the bone, intraosseous fluid flow occurs through the interconnected pore space in the bone. Bone fluid flow leads to a strain generated streaming potential (SGP). However, there is no experimental study on the relationship between the generation of intraosseous pressure and the SGP. The purpose of this study was to obtain the relationship between SGP and intraosseous pressure generations in cortical bone. In order to understand the issue, a drained, one-dimensional experimental setup for fluid filled cortical bone samples with four different strain rates was used to simultaneously measure the intraosseous pressure and SGP. The results revealed a significant correlation (r = 0.98, p = 0.02) between the generation of the SGP and the intraosseous pressure, which indicates that an intraosseous pressure gradient produces a SGP in cortical bone. PMID- 17914631 TI - Experimental composites made of electron beam irradiated reinforced fillers. AB - This study investigated the influence of electron beam irradiated reinforced fillers on the three body wear and flexural strength of experimental composite blends. Three formulations of reinforced fillers were investigated: (A) high loaded inorganic filler composite with 60 wt.% SiO2, (B) low loaded inorganic filler composite with 40 wt.% SiO2, (C) organic filler composite (precipitated Bis-phenol-A-di-methacrylate). The fillers were assigned to two subgroups of unswollen (A, B, C) and monomer swollen (As, Bs, Cs) fillers. The experimental blends (matrix: Urethane-dimethacrylate) were mixed using un-treated, annealed (90 degrees C), or electron beam irradiated fillers with 30 and 90 kGy, respectively. All specimens were heat-cured for 20 min at 140 degrees C. Three body abrasion and flexural strength tests were performed. The highest flexural strength was evaluated for composites made of the 30 kGy irradiated type Bs filler. The comparison with annealed fillers showed that the effect was independent of increasing temperatures during the radiation process. Blends with a SiO2 content of 60 wt.% (type A, As) had significantly less wear than blends with 40 wt.% (type B, Bs) or blends with organic fillers (type C, Cs). The flexural strength of the composite could be improved by using pre-irradiated reinforced fillers. However, wear was not affected using this procedure. PMID- 17914632 TI - Comparative in vitro study of the proliferation and growth of ovine osteoblast like cells on various alloplastic biomaterials manufactured for augmentation and reconstruction of tissue or bone defects. AB - In this in vitro study ovine osteoblast-like cells were cultured on seven different alloplastic biomaterials used for augmentation and for reconstruction of bone defects in dental and craniomaxillofacial surgery. The aim of this study was to examine the growth behaviour (viability, cell density and morphology) of ovine osteoblast-like cells on the investigated biomaterials to get knowledge which biomaterial is qualified to act as a cell carrier system in further in vivo experiments. The biomaterials were either synthetically manufactured or of natural origin. As synthetically manufactured biomaterials Ethisorb, MakroSorb, PalacosR, and PDS film were used. As biomaterials of natural origin BeriplastP, Bio-Oss and Titanmesh were investigated. The cell proliferation and cell colonization were analyzed by a proliferation assay and scanning electron microscopy. Osteoblast-like cells proliferated and attached on all biomaterials, except on Beriplast. On Ethisorb the highest cell proliferation rate was measured followed by PalacosR. Both biomaterials offer suitable growth and proliferation conditions for ovine osteoblast-like cells. The proliferation rates of Bio-Oss, MakroSorb, PDS-film and Titanmesh were low and SEM examinations of these materials showed less spread osteoblast-like cells. The results showed that ovine osteoblast-like cells appear to be sensitive to substrate composition and topography. This in vitro study provides the basis for further in vivo studies using the sheep model to examine the biocompatibility and the long-term interaction between the test material and tissue (bone regeneration). PMID- 17914633 TI - Behaviors of keratinocytes and fibroblasts on films of PLA50-PEO-PLA50 triblock copolymers with various PLA segment lengths. AB - The growth of human primary keratinocytes and fibroblasts on PLA-PEO-PLA copolymer films was investigated as an intermediate stage of a strategy aimed at making implantable dermo-epidermal substitutes. Four PLA-PEO-PLA triblock copolymers with the same PEO block and different DL-lactic acid/ethylene oxide molar ratios (LA/EO) (0.8, 1.4, 1.8 and 2), were synthesized and characterized by 1H-nuclear magnetic resonance and infrared spectroscopy. The films made of these copolymers were more hydrophilic than PLA50 and than tissue culture polystyrene controls according to contact angles with water. Proliferation and adhesion of human skin cells were evaluated by MTT assay and by scanning electron microscopy. The presence of PEO in the triblock copolymers influenced cell adhesion and proliferation of fibroblasts, whereas keratinocyte adhesion and proliferation were not affected. These features emphasize the interest of PLA-PEO-PLA triblock copolymers to serve as better compounds than the racemic PLA previously investigated to make supports for human skin primary cells and scaffolds for skin engineering. PMID- 17914634 TI - Cell type-specific aspects in biocompatibility testing: the intercellular contact in vitro as an indicator for endothelial cell compatibility. AB - Endothelial cells cover the inner surface of blood vessels and form the interface between the blood and the tissues. Endothelial cells are involved in regulating barrier function, which is maintained by the interendothelial cell contacts. These interendothelial cell contacts are established by the interaction of different molecules. The maintenance of the barrier requires an appropriate signalling between these molecules. Thus, a number of different signalling pathways are integrated within interendothelial contacts. Since endothelial cells are important in tissue-implant interactions (especially for stent materials) this study examines the expression pattern of different interendothelial contact molecules to determine the usefulness in the analysis of biocompatibility in vitro. The effects of different pro-inflammatory and toxic stimuli and contact of human microvascular endothelial cells to metallic surfaces were examined for their impact on the pattern of interendothelial contact molecules. Striking modifications in the arrangement of these molecules were induced and the mode of modification was dependent on the tested compound. Thus, examining the pattern of expression of specific interendothelial contact molecules in vitro may be useful for testing the endothelial cell compatibility of biomaterials and their corrosion products. PMID- 17914635 TI - Effects of bioactive ceramics on the pathogenesis of rat vascular smooth muscle cells treated with phorbol 12-myristate 13-acetate. AB - Vascular smooth muscle cells (VSMCs) play a pivotal role in vascular injury through proliferation and migration. Pro-inflammatory cytokines and cyclooxygenase (COX)-2 and nitric oxide synthase (NOS) are highly associated with the pathogenesis of VSMCs. We investigated the effect of bioactive ceramics on the expression of inflammatory cytokines, COX-2, and inducible NOS (iNOS) induced by phorbol 12-myristate 13-acetate (PMA) in rat VSMCs. The ceramics inhibited mRNA expression of IL-1beta, TNF-alpha, IL-6, COX-2, and iNOS. Prostaglandin release was also diminished by the ceramics. The bioactive ceramics effect on cytokines, COX-2, and iNOS expression was achieved by inhibition of NF-kappaB activity. Interestingly, the ceramics-induced up-regulation of expression of endothelial NOS resulted in an increase of nitric oxide production. Thus, bioactive ceramics may have dual effects on the pathogenesis of VSMCs by regulation of NF-kappaB activity and NO production. PMID- 17914636 TI - Preparation and function of composite asymmetric chitosan/CM-chitosan membrane. AB - A novel composite asymmetric chitosan/CM-chitosan membrane (C-P-C) was prepared, the top-layer was chitosan (CS), the intermediate was PVA, and the substrate was carboxymethyl chitosan (CM-CS). C-P-C membrane had capability in mechanical strength, light transparence, vapor permeability, and wound skin joining. The CS and CM-CS in C-P-C membrane were selected by series independent experiments, respectively. CS (MW 90,000 Da) had the highest antibacterial activity for E.coli. CM-CS had biocompatibility, no cytotoxicity, and had the activity of promoting growth of human skin fibroblast and inhibiting the growth of keloid fibroblast. The normal skin fibroblast can growth on the CM-CS surface of C-P-C, and have no conglomeration in higher cell density, and the keloid fibroblast could not growth on CM-CS surface of C-P-C. The animal experiment demonstrated that wound, covered with the C-P-C membrane, was hemostatic, healing quickly and had histocompatibility. The results indicated that the C-P-C membrane could be used as dressing of skin repair, and had the potential in promoting wound healing and inhibiting the keloid formation. PMID- 17914637 TI - Self-assembly and bioactive response of a crystalline metal oxide in a simulated blood fluid. AB - In this study we report on the bioactive response of self-assembled niobium oxide microstructures when immersed in a supersaturated solution emulating mineral content in blood. The structures were formed via electrochemical anodization in an electrolyte comprised of HF and NaF. The slow oxide formation kinetics associated with the presence of NaF in the electrolyte enabled microscopic examinations during microstructure evolution as shown via scanning electron microscopy (SEM). Apparently the slow growth kinetics encourage the development of bioactive sites on the microstructures, as these structures induced mineral formations. On the other hand, microstructures grown in the absence of salt were ineffective mineral nucleators. Analysis of nucleated mineral deposits was performed using X-ray diffraction and Raman spectroscopy. Both long-range and short-range order experiments verified the nucleated mineral phase was hydroxyapatite (HAP). Further characterization of the mineral phase was observed using SEM and revealed effective nucleation sites were predominantly isolated to loci on the ordered microbodies as opposed to locations lying within the amorphous strata. PMID- 17914638 TI - Generation of hydroxyapatite patterns by electrophoretic deposition. AB - Hydroxyapatite (HAp) patterns with distinct boundaries were generated by electrophoretic deposition (EPD) utilizing an insulating mask that partially blocks the electric field. For the EPD process, we selected two types of mask: a polytetrafluoroethylene (PTFE) board with holes and a resist pattern. A porous PTFE film, which differed from the mask PTFE, was employed as a substrate and attached to the mask. EPD was performed with a suspension of wollastonite particles in acetone, which were deposited on the substrate in the form of the patterned mask. The deposited wollastonite particles induced HAp patterns during a soak in simulated body fluid (SBF). As a result, minute HAp patterns, such as dots, lines, and corners were fabricated on the porous PTFE substrate with a minimum line width of about 100 microm. PMID- 17914640 TI - Murine fertilized ovum, blastomere and morula cells lacking SP phenotype. AB - In the field of stem cell research, SP (side population) phenotype is used to define the property that cells maintain a high efflux capability for some fluorescent dye, such as Hoechst 33342. Recently, many researches proposed that SP phenotype is a phenotype shared by some stem cells and some progenitor cells, and that SP phenotype is regarded as a candidate purification marker for stem cells. In this research, murine fertilized ova (including conjugate and single nucleus fertilized ova), 2-cell stage and 8-cell stage blastomeres, morulas and blastocysts were isolated and directly stained by Hoechst 33342 dye. The results show that fertilized ovum, blastomere and morula cells do not demonstrate any ability to efflux the dye. However, the inner cell mass (ICM) cells of blastocyst exhibit SP phenotype, which is consistent with the result of embryonic stem cells (ESCs) in vitro. These results indicate that the SP phenotype of ICM-derived ESCs is an intrinsic property and independent of the culture condition in vitro, and that SP phenotype is one of the characteristics of at least some pluripotent stem cells, but is not shared by totipotent stem cells. In addition, the result that the SP phenotype of ICM cells disappeared when the inhibitor verapamil was added into medium implies that the SP phenotype is directly associated with ABCG2. These results suggest that not all the stem cells demonstrate SP phenotype, and that SP phenotype might act as a purification marker for partial stem cells such as some pluripotent embryonic stem cells and multipotent adult stem cells, but not for all stem cells exampled by the totipotent stem cells in the very early stage of mouse embryos. PMID- 17914641 TI - Involvement of nitric oxide signaling in mammalian Bax-induced terpenoid indole alkaloid production of Catharanthus roseus cells. AB - Bax, a mammalian pro-apoptotic member of the Bcl-2 family, has been demonstrated to be a potential regulatory factor for plant secondary metabolite biosynthesis recently. To investigate the molecular mechanism of Bax-induced secondary metabolite biosynthesis, we determined the contents of nitric oxide (NO) of the transgenic Catharanthus roseus cells overexpressing a mouse Bax protein and checked the effects of NO specific scavenger 2,4-carboxyphenyl-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide (cPITO) on Bax-induced terpenoid indole alkaloid (TIA) production of the cells. The data showed that overexpression of the mouse Bax in C. roseus cells triggered NO generation of the cells. Treatment of cPITO not only inhibited the Bax-triggered NO burst but also suppressed the Bax-induced TIA production. The results indicated that the mouse Bax might activate the NO signaling in C. roseus cells and induce TIA production through the NO-dependent signal pathway in the cells. Furthermore, the activities of nitric oxide synthase (NOS) were significantly increased in the transgenic Bax cells as compared to those in the control cells, showing that the mouse Bax may induce NOS of C. roseus cells. Treatment of the transgenic Bax cells with NOS inhibitor PBITU blocked both Bax-induced NO generation and TIA production, which suggested that the mouse Bax might trigger NO generation and TIA production through NOS. However, the NOS-like activities and NO generation in the transgenic Bax cells did not match kinetically and the Bax-induced NOS-like activity was much later and lower than NO production. Moreover, the Bax-induced NO generation and TIA production were only partially inhibited by PBITU. Thus, our results suggested that the Bax-induced NO production and secondary metabolite biosynthesis in C. roseus cells was not entirely dependent on NOS or NOS-like enzymes. PMID- 17914639 TI - Implants of polyanionic collagen matrix in bone defects of ovariectomized rats. AB - In recent years, there has been a great interest in the development of biomaterials that could be used in the repair of bone defects. Collagen matrix (CM) has the advantage that it can be modified chemically to improve its mechanical properties. The aim of the present study was to evaluate the effect of three-dimensional membranes of native or anionic (submitted to alkaline treatment for 48 or 96 h) collagen matrix on the consolidation of osteoporosis bone fractures resulting from the gonadal hormone alterations caused by ovariectomy in rats subjected to hormone replacement therapy. The animals received the implants 4 months after ovariectomy and were sacrificed 8 weeks after implantation of the membranes into 4-mm wide bone defects created in the distal third of the femur with a surgical bur. Macroscopic analysis revealed the absence of pathological alterations in the implanted areas, suggesting that the material was biocompatible. Microscopic analysis showed a lower amount of bone ingrowth in the areas receiving the native membrane compared to the bone defects filled with the anionic membranes. In ovariectomized animals receiving anionic membranes, a delay in bone regeneration was observed mainly in animals not subjected to hormone replacement therapy. We conclude that anionic membranes treated with alkaline solution for 48 and 96 h presented better results in terms of bone ingrowth. PMID- 17914642 TI - Evolution of cd59 gene in mammals. AB - The CD59-coding sequences were obtained from 5 mammals by PCR and BLAST, and combined with the available sequences in GenBank, the nucleotide substitution rates of mammalian cd59 were calculated. Results of synonymous and nonsynonymous substitution rates revealed that cd59 experienced negative selection in mammals overall. Four sites experiencing positive selection were found by using "site specific" model in PAML software. These sites were distributed on the molecular surface, of which 2 sites located in the key functional domain. Furthermore, "branch-site-specific" model detected 1 positive site in cd59a and cd59b lineages which underwent accelerated evolution caused by positive selection after gene duplication in mouse. PMID- 17914644 TI - Comparative studies on the structure and adhesion of setae in G. gecko and G. swinhonis. AB - Scanning electron microscopy (SEM) and histological techniques were used to observe and study the setae structures of two gecko species (G. gecko and G. swinhonis) and the relationships between these structures and the adhesive forces. The SEM results showed that the setae of these two species were densely distributed in an orderly fashion, and branched with curved tips. The setae of G. gecko had cluster structures, each cluster containing 4-6 setae whose terminal branches curved towards the center of the toes at approximately 10 degrees , the tips of the branches like spatulae and densely arrayed at an interval of less than 0.2-0.3 microm. On the contrary, the branch tips in the setae of G. swinhonis were curled, and the terminal parts of setae curved towards the center of the toes at various angles. Usually the setae of these gecko species branch twice at the top at intervals greater than that of G. gecko. The histological observation found that inside the setae of these two species there were plenty of unevenly distributed contents, such as epithelia, fat cells, pigmental cells and muscle tissue, but no gland cells existed. The results of functional experiments suggested that modifying the structure of gecko's setae could reduce its adhesive ability dramatically, demonstrating the positive correlation between the structure of the gecko's setae and its adhesive ability. The above results provide important information in designing bio-mimic setae and bio-gecko robots. PMID- 17914643 TI - 3D-QSAR analysis of a new type of acetylcholinesterase inhibitors. AB - Acetylcholinesterase (AChE) inhibitors are an important class of medicinal agents used for the treatment of Alzheimer's disease. A screening model of AChE inhibitor was used to evaluate the inhibition of a series of phenyl pentenone derivatives. The assay result showed that some compounds displayed higher inhibitory effects. In order to study the relationship between the bioactivities and the structures, 26 compounds with phenyl pentenone scaffold were analyzed. A 3D-QSAR model was constructed using the method of comparative molecular field analysis (CoMFA). The results of cross-validated R(2)cv=0.629, non-cross validated R(2)=0.972, SE=0.331, and F=72.41 indicate that the 3D-model possesses an ability to predict the activities of new inhibitors, and the CoMFA model would be useful for the future design of new AChE inhibitors. PMID- 17914645 TI - In vitro insulin refolding: characterization of the intermediates and the putative folding pathway. AB - The in vitro refolding process of the double-chain insulin was studied based on the investigation of in vitro single-chain insulin refolding. Six major folding intermediates, named P1A, P2B, P3A, P4B, P5B, and P6B, were captured during the folding process. The refolding experiments indicate that all of these intermediates are on-pathway. Based on these intermediates and the formation of hypothetic transients, we propose a two-stage folding pathway of insulin. (1) At the early stage of the folding process, the reduced A chain and B chain individually formed the intermediates: two A chain intermediates (P1A and P3A), and four B chain intermediates (P2B, P4B, P5B, and P6B). (2) In the subsequent folding process, transient I was formed from P3A through thiol/disulfide exchange reaction; then, transients II and III, each containing two native disulfides, were formed through the recognition and interaction of transient I with P4B or P6B and the thiol group's oxidation reaction mainly using GSSG as oxidative reagent; finally, transients II and III, through thiol/mixture disulfide exchange reaction, formed the third native disulfide of insulin to complete the folding. PMID- 17914646 TI - Association between depressive mood and cigarette smoking in a large Italian sample of smokers intending to quit: implications for treatment. AB - OBJECTIVE: The aim of the study is to assess the prevalence and degree of depression at baseline of a large cohort of smokers intending to quit. METHODS: A cross-sectional investigation was carried out on a population of 757 smokers attending the Medical Service for Addictive Disorders, at Verona University Hospital. The degree of nicotine addiction was measured by the Fagerstrom Test of Nicotine Dependence (FTND) and current mood tested by the Self-rating Depression Scale (SDS), a commonly used and well validated instrument to assess depressive mood. RESULTS: Two hundred and twenty-two subjects (30.3%) were depressed at baseline (SDS test score > or = 50). Bivariate analysis, using the SDs score dichotomised at the cut-off of 50 as dependent variable, shows that female gender (p=0.01) and widowhood (p<0.001) were correlated to depression. Logistic regression analysis confirms the correlation between depression and female gender (OR=2.03, IC 95%=1.42-2.88, p<0.001) and between depression and widowhood, with the greatest risk of depression among widows and widowers (OR=3.22, IC 95%=1.01 10.27, p<0.048). CONCLUSIONS: The study showed a high degree of pre-treatment depression in smokers intending to quit. Although the association between depression and nicotine dependence has been consistently reported many times, and it is well known that depressed subjects find it more difficult to quit, most guidelines seem not to consider this connection. These findings suggest the need for baseline assessment of depression by screening all smokers seeking assistance in quitting, a priority health objective because smoking is the number one avoidable killer in developed countries. PMID- 17914647 TI - Clinical outcome of long-term home parenteral nutrition in non-oncological patients: a report from two specialised centres. AB - OBJECTIVE AND METHODS: Forty-one (20 m, 21 f) non-oncological home parenteral nutrition (HPN) patients (52.0+/-16.6 years, BMI 20.2+/-4.0 kg/m(2)), enrolled from 1995 to 2005, underwent a clinical and biochemical follow-up at 3 months, 1 and 3 years. RESULTS: At admittance the Karnofsky Index ranged between 40 and 50 in 13 (31.7%) and 60 and 80 in 28 (68.3%) patients; the most frequent underlying disease was mesenteric infarction (11 patients, 27%). All had a central venous access. Mean catheterization days were 864+/-992, while mean HPN days were 630+/ 668. At the 3-month follow-up, all patients were on HPN, at 1 year 24 (58.5%) and at 3 years 11 (27%). At 3 months, 1 and 3 years there was a significant increase in BMI (p=0.001), body weight (p=0.001) and Karnofsky Index (p=0.001), as well as an improvement of several biochemical parameters. Survival rate of HPN patients was 90.2% at 1 year, 87.8% at 3 years and 82.9% at 5 years. As to HPN-related complications, there was a central venous catheter (CVC) obstruction in 8 patients (19.5%) and 47 CVC infections in 24 (58.5%) patients. The infection rate was 1.32/(00) days of catheterization (1.8/(00) from 1995 to 1998 and 1.0/(00) from 1999 to 2005). Hospitalisation was necessary in over 50% of patients, and death occurred in 8, always as a consequence of the primary disease. CONCLUSIONS: The experience of the nutritional team and careful patient and caregiver training reduce CVC infection rate and the overall risk of complications possibly due to HPN. PMID- 17914649 TI - Adaptations for vision in dim light: impulse responses and bumps in nocturnal spider photoreceptor cells (Cupiennius salei Keys). AB - The photoreceptor cells of the nocturnal spider Cupiennius salei were investigated by intracellular electrophysiology. (1) The responses of photoreceptor cells of posterior median (PM) and anterior median (AM) eyes to short (2 ms) light pulses showed long integration times in the dark-adapted and shorter integration times in the light-adapted state. (2) At very low light intensities, the photoreceptors responded to single photons with discrete potentials, called bumps, of high amplitude (2-20 mV). When measured in profoundly dark-adapted photoreceptor cells of the PM eyes these bumps showed an integration time of 128 +/- 35 ms (n = 7) whereas in dark-adapted photoreceptor cells of AM eyes the integration time was 84 +/- 13 ms (n = 8), indicating that the AM eyes are intrinsically faster than the PM eyes. (3) Long integration times, which improve visual reliability in dim light, and large responses to single photons in the dark-adapted state, contribute to a high visual sensitivity in Cupiennius at night. This conclusion is underlined by a calculation of sensitivity that accounts for both anatomical and physiological characteristics of the eye. PMID- 17914648 TI - Stress and cocaine interact to modulate basic fibroblast growth factor (FGF-2) expression in rat brain. AB - RATIONALE: Our laboratory has previously demonstrated that the expression of basic fibroblast growth factor (FGF-2), a protein involved in survival and maintenance of several cell phenotypes as well as in synaptic plasticity, is modulated by stress (Molteni et al., Brain Res Rev 37:249-258, 2001; Fumagalli et al., Neurobiol Dis 20:731-737, 2005) and cocaine (Fumagalli et al., J Neurochem 96:996-1004, 2006). OBJECTIVES: Since it is widely recognized that stress influences drug seeking, we decided to investigate whether stress, acute or repeated, could influence the changes in FGF-2 gene expression brought about by cocaine. RESULTS: Our data demonstrate that stress and cocaine interact to produce significant changes on FGF-2 expression in rat prefrontal cortex and striatum. In prefrontal cortex, our experiments demonstrated that a single exposure to stress potentiated cocaine-induced FGF-2 elevation, whereas prolonged stress prevented the modulation of the trophic factor in response to cocaine. In striatum, the magnitude of cocaine-induced FGF-2 response is enhanced by repeated stress, whereas no interaction was observed when acute stress and single exposure to cocaine were combined. CONCLUSIONS: Our findings demonstrate that stress interacts with cocaine to alter the pattern of FGF-2 expression in a way that depends on whether stress is acute or chronic and in a regionally selective fashion. These results identify a potential molecular target through which stress alters cellular sensitivity to cocaine and might prove useful in understanding the mechanisms underlying brain vulnerability to stress. PMID- 17914650 TI - Invasion of Rh Null Cells by Plasmodium falciparum identifies a new invasion pathway. AB - The malaria parasite, Plasmodium falciparum, invades the human erythrocyte through a complex interaction with erythrocyte receptors characterized by patterns of resistance to various enzymes. As invasion rates are influenced by blood group polymorphisms, we reasoned that the extremely rare rhesus null (Rh(null)) erythrocytes could be informative in characterizing receptors. The aim was to test whether the complete absence of the Rh complex from the cell membrane impacted on parasite invasion. Enzyme treatment patterns for four P. falciparum isolates were first characterised for normal Rh cells. Two isolates showed an enzyme treatment pattern not hitherto described, with resistance to neuraminidase, trypsin and chymotrypsin. In contrast, all isolates had enhanced invasion rates for the Rh(null) cell for all enzyme treatment regimens. The first finding suggests there is another pathway that P. falciparum can utilise to invade the host. We speculate that the Rh null cell membrane exposes a novel ligand defined as Receptor N. PMID- 17914652 TI - Efficacy of single-session percutaneous drainage and 50% acetic Acid sclerotherapy for treatment of simple renal cysts. AB - PURPOSE: To evaluate the efficacy and long-term results of single-session 50% acetic acid sclerotherapy for the treatment of simple renal cysts, and to compare the therapeutic results of 5 and 20 min sclerosant dwell techniques. METHODS: During the past 9 years, 50% acetic acid sclerotherapy was performed on 67 cysts in 66 patients. An acetic acid volume corresponding to a mean of 23% of the aspirated cyst volume was injected into the cysts. A 20 min dwell time with position changes was performed in 32 cysts (31 patients; group I) and 8% of volume for a 5 min dwell time in 35 cysts (35 patients; group II). Three- and 6 month sonographic or CT follow-up was performed for a minimum of 1 year. Complete regression was defined as no remaining cyst measurable on sonography with or without a scar at the renal cortex. Partial regression was defined as a decreased cyst volume compared with that before sclerotherapy. The Mann-Whitney U-test was used to compare the therapeutic results between the two groups. RESULTS: For 67 simple renal cysts, complete regression on follow-up was observed in 21 of 32 cysts (66%; group I) and 22 of 35 cysts (63%; group II); the remaining 24 cysts all showed partial regression. The partial reduction rate of the cyst's volume was 97.4% (91.3-99.4%) in group I and 96.9% (90.8-99.5 %) in group II. There were no procedure-related major complications, and no statistically significant differences in the complete regression and partial volume reduction rates between the two groups (p > 0.05). CONCLUSION: Fifty percent acetic acid is an effective and safe sclerosing agent for simple renal cysts. Fifty percent acetic acid sclerotherapy with a 5 min sclerosant dwell time, using a volume of about 10% of the aspirated volume, is sufficient for satisfactory results of simple renal cyst sclerotherapy. PMID- 17914651 TI - Correlation of CK-20-positive cells in peripheral venous blood with serum CEA levels in patients with colorectal carcinoma. AB - Tumor cell dissemination appears to be an early event in tumor progression, and tumor cells can be detected in peripheral venous blood at the time of the operation. Although cytokeratin 20 (CK-20) is not specifically expressed by colorectal carcinomas, it represents a widely used marker for the detection of colorectal tumor cells. We used the combination of density centrifugation and CK 20 real-time reverse transcription polymerase chain reaction to detect CK-20 positive cells in the peripheral venous blood of 37 patients with colorectal carcinoma. Detection rates were compared to serum levels of the tumor markers carcinoembryonic antigen (CEA), carbohydrate antigen CA 19-9, and cancer antigen CA 125. The prognostic impact was assessed by the overall survival and by univariate and multivariate analysis. Overall, CK-20-positive cells in peripheral venous blood were detected in 11 of 37 (29.7%) patients. CK-20-positive patients showed a significantly higher mean serum CEA level (90.3 ng/ml) than the 4.1 ng/ml found in the CK-20-negative group (p = 0.03). CEA levels also correlated with CK-20 copy numbers. No significant correlation was observed for CA 19-9 or CA 125. CK-20-negative patients showed a trend toward better survival (p = 0.08). In the univariate analysis, CA 19-9, CEA, tumor size, lymph node status, grading, the presence of distant metastases, and resection status reached significant prognostic levels, whereas the detection of CK-20-positive cells showed only a prognostic trend (p = 0.06). Multivariate analysis failed to identify independent prognostic parameters. Here we report the correlation of CK-20-positive cells in peripheral venous blood with the serum CEA level of patients with colorectal cancer, which may represent a potential marker of the tumor load. PMID- 17914653 TI - Continuous wound infusion of local anesthetic for the control of pain after elective abdominal colorectal surgery. AB - PURPOSE: Local anesthetic wound infusion has been investigated in recent years as a potential alternative to standard analgesic regimens after major surgery. This study investigates the efficacy of a continuous wound infusion of ropivacaine in conjunction with best practice postoperative analgesia after midline laparotomy for abdominal colorectal surgery. METHODS: We performed a randomized, participant and outcome assessor-blinded, placebo-controlled trial on patients presenting for major abdominal colorectal surgery at our institution between December 2003 and February 2006. Patients were allocated to receive ropivacaine 0.54 percent or normal saline via a dual catheter Painbuster Soaker (I-Flow Corporation, OH, USA) continuous infusion device into their midline laparotomy wound for 72 hours postoperatively. RESULTS: A total of 310 patients were included in this study. The continuous wound infusion of ropivacaine after abdominal colorectal surgery conveys minimal benefit compared with saline wound infusion. No statistically significant difference could be shown for: pain at rest, morphine usage, length of stay, mobility, nausea, or return of bowel function. There was a small, statistically significant difference in mean pain on movement on Day 1 for the ropivacaine group (adjusted mean difference -0.6 (range, -1.08 to -0.13)). Although this trend continued on Days 2 and 3, the differences between groups were no longer statistically significant. CONCLUSIONS: Management of pain after major abdominal colorectal surgery is best achieved through adopting a multimodal approach to analgesia. Delivery of ropivacaine to midline laparotomy wounds via a Painbuster Soaker device is safe, but we have not demonstrated any significant clinical advantage over current best practice. PMID- 17914654 TI - Biofeedback vs. electrostimulation in the treatment of postdelivery anal incontinence: a randomized, clinical trial. AB - PURPOSE: This study was designed to evaluate the effect of biofeedback and electrostimulation in a randomized, clinical trial for the treatment of patients with postdelivery anal incontinence. METHODS: Forty-nine females who sustained third-degree or fourth-degree perineal rupture with a mean age of 36 (range, 22 44) years were included in the study. The females were randomized to biofeedback or electrostimulation treatment. Forty females completed the study: 19 in the biofeedback and 21 in the electrostimulation group. Biofeedback or electrostimulation sessions were performed two times daily for eight weeks in each group. Wexner incontinence score, fecal incontinence quality of life scores, and reduced quality of life on visual analog scale were registered before and after treatment. Patients' self-rating of treatment effect also was registered in both groups. The primary outcome measure was the Wexner incontinence score. RESULTS: There were no differences in treatment effect between groups. Comparing pretreatment status to posttreatment in each group showed no improvement in Wexner score, reduced quality of life, or any of the fecal incontinence quality of life scores. Patients' self-rating of the treatment effect, however, showed a subjective improvement of symptoms both in the biofeedback and in the electrostimulation group (median, 7 vs. 5.) CONCLUSIONS: This study shows that there was no difference in effect between biofeedback and electrostimulation. Neither biofeedback nor electrostimulation treatments improved Wexner incontinence score, reduced quality of life, or fecal incontinence quality of life scores. Both treatments resulted in improvement of patients' subjective perception of incontinence control. PMID- 17914655 TI - Earlier eradication of intra-anal warts with argon plasma coagulator combined with imiquimod cream compared with argon plasma coagulator alone: a prospective, randomized trial. AB - PURPOSE: Despite the increasing incidence of condylomata acuminate, optimal treatment of anal warts is still undecided. This prospective, randomized study was designed to compare the efficacy of combined argon plasma coagulation and imiquimod cream vs. argon plasma coagulation alone in the management of intra anal warts. METHODS: From October 2002 to March 2005, 49 patients with intra-anal warts were randomly assigned to argon plasma coagulation plus imiquimod cream (n = 24) vs. argon plasma coagulation alone (n = 25). Therapeutic sessions were repeated until the elimination of the warts. Efficacy of therapy was defined as the time needed for eradication. All patients were followed up for a mean period of 12 months for signs of recurrence. RESULTS: Elimination of warts was achieved earlier in patients receiving combination therapy compared with those receiving monotherapy with argon plasma coagulation (62.5 +/- 5.4 days vs. 91.2 +/- 6.4 days; P = 0.0016). A subgroup analysis performed in HIV-positive patients showed similar results (combination therapy 95 +/- 22.6 days; monotherapy 124.3 +/- 20.7 days; P = 0.033); however, in HIV-positive patients warts were eradicated later compared with HIV-negative patients (110.8 +/- 25.7 days vs. 65 +/- 25.4 days; P < 0.0001). No major complications were observed in our study population. After the follow-up period, recurrence of warts was evident in 22.7 percent of patients in the combination group compared with 34.7 percent of patients in the monotherapy group (P = 0.51). Recurrence was significantly higher in HIV-positive patients compared with HIV-negative patients (P = 0.0039). CONCLUSIONS: Combination therapy with argon plasma coagulator plus imiquimod cream results in earlier clearance of intra-anal warts in both immunocompetent and immunocompromised patients; however, it does not affect the rate of recurrence. PMID- 17914656 TI - The exact distribution of divergence times. AB - The recent work of Haubold and Wiehe (Mol. Biol. Evol. 18:1157-1160, 2001) considered statistical inference of the divergence time. However, there appears to be a fundamental flaw in the paper since it treated the divergence time as a random variable and not as a parameter. In this note, we derive a valid statistical inference for the divergence time. We derive an estimator for the divergence time as well as explicit expressions for its the probability density function, cumulative distribution function and the means. We also provide a 5 line computer program for computing the associated confidence intervals. We expect that the results presented could be useful for statistical modeling of divergence times. PMID- 17914657 TI - Sensory irritation response in rats: modeling, analysis and validation. AB - Inhaled gases can cause respiratory depression by irritating (stimulating) nerves in the nasal cavity. Respiratory depression, in turn, decreases the rate of delivery of those gases to the stimulated nerves, potentially leading to a complex feedback response. In order to better understand how the nervous system responds to such chemicals, a mathematical model is created to describe how the presence of irritants affects respiration in the rat. The ordinary differential equation model describes the dosimetry of these reactive gases in the respiratory tract, with particular focus on the physiology of the upper respiratory tract, and on the neurological control of respiration rate due to signaling from the irritant-responsive nerves in the nasal cavity. The ventilation equation is altered to account for an apparent change in dynamics between the initial ventilation decrease and the recovery to steady state as seen in formaldehyde exposure data. Further, the model is evaluated and improved through optimization of particular parameters to describe formaldehyde-induced respiratory response data and through sensitivity analysis. The model predicts the formaldehyde data well, and hence the model is thought to be a reasonable description of the physiological system of sensory irritation. The model is also expected to translate well to other irritants. PMID- 17914658 TI - Rapid measurement of 3J(H N-H alpha) and 3J(N-H beta) coupling constants in polypeptides. AB - We present two NMR experiments, (3,2)D HNHA and (3,2)D HNHB, for rapid and accurate measurement of 3J(H N-H alpha) and 3J(N-H beta) coupling constants in polypeptides based on the principle of G-matrix Fourier transform NMR spectroscopy and quantitative J-correlation. These experiments, which facilitate fast acquisition of three-dimensional data with high spectral/digital resolution and chemical shift dispersion, will provide renewed opportunities to utilize them for sequence specific resonance assignments, estimation/characterization of secondary structure with/without prior knowledge of resonance assignments, stereospecific assignment of prochiral groups and 3D structure determination, refinement and validation. Taken together, these experiments have a wide range of applications from structural genomics projects to studying structure and folding in polypeptides. PMID- 17914659 TI - Iliac artery stenosis as a cause of posttransplant renal failure and claudication. AB - Iliac artery stenosis (IAS) is a rare complication after renal transplantation. We demonstrate a case of ipsilateral external IAS proximally to anastomosis in a kidney recipient, which manifested with renal failure and claudication, and was successfully treated with endovascular stent placement. PMID- 17914661 TI - Roles of Medicines Counter Assistants in advice giving in community pharmacies: a Discourse Analysis. AB - OBJECTIVE: Research in interaction and communication in community pharmacies has been reported in a number of countries but to date, talk at the micro-level in the community pharmacy context in the UK has received virtually no study. The objective of this exploratory study was to identify what light a Discourse Analytic approach could shed to describe the role(s) of Medicines Counter Assistants (MCAs) when dealing with clients relating to health, illness or medicines. METHODS: Discourse Analysis is a growing field of study which has its roots in linguistics, but also in critical theory, philosophy, sociology, psychology and more. It is used in various ways by social scientists studying spoken and written language in use. Following research ethics approval and informed consent from participants, audio-recordings of staff-client interactions in three community pharmacies in Wales, UK were transcribed and analysed from a Discourse Analytic perspective, focussing on the emerging role of the MCA in the negotiation of advice. Main outcome measure Transcripts of interactions in community pharmacies analysed using Discourse Analysis. RESULTS: Ten hours of recordings were made. Of the 41 recorded interactions obtained, 29 involved an MCA regarding health and/or medicines. Example extracts represent different ways in which MCAs are involved in the interactions, ranging from dealing solely with the client, to the MCA dealing with the client but checking advice with the pharmacist, the MCA acting as a gatekeeper, the pharmacist intervening in the MCA client interaction, the MCA keeping the client 'on hold' until the pharmacist is ready to interact with them and the MCA acting as an active intermediary. With the differing involvement, slightly different institutional and participant roles emerge for the MCAs through talk and interaction. CONCLUSION: This exploratory study succeeded in producing transcripts of interactions between MCAs, and clients and indicated that Discourse Analysis is helpful in assisting our understanding of the involvement of MCAs in UK community pharmacy encounters. With regards the negotiation of advice, we have shown how the physical and legal contexts, in addition to the discursive/interactional context, play a part in information delivery, as does the different staff's access to knowledge. PMID- 17914660 TI - Sleep disorders: a systematic review of an emerging major clinical issue in renal patients. AB - The prevalence of sleep disorders is significantly higher (up to 80%) in patients with chronic uremia compared to the general population. Sleep disorders appear even in the early stages of chronic kidney disease. These disturbances are complex, including difficulties in falling asleep and awakening, interrupted sleep, nightmares, restless legs syndrome, sleep apnea syndrome, etc. There are still disagreements on the major etiological factors of sleep disorders in the uremic patient. Older age, long dialysis vintage, alcohol and tobacco abuse and, particularly, the presence of significant comorbidities are major determinants of sleep disorders in dialysis patients. Proper assessment of sleep disorders in the renal population is still under investigation; recent studies have mostly addressed patients' perception based on questionnaires. More precise polysomnographic assessments are less studied in renal patients. Sleep disorders significantly affect quality of life in dialysis patients. An accurate and early identification of such disturbances would lead to a significant improvement in quality of life, and probably also in outcome, in uremic patients. Sleep apnea syndrome is extremely frequent in dialysis patients, with obvious consequences for cardiovascular morbidity and mortality. Proper diagnosis and therapy of sleep apnea syndrome could significantly reduce cardiovascular risk. Although sleep quality improves after renal transplantation, allograft recipients still have significantly more sleep disorders than healthy individuals. Here, we review recent data on sleep disturbances in renal patients, focusing on the end-stage renal disease patient treated by dialysis. PMID- 17914662 TI - Paracoccidioidomycosis in wild monkeys from Parana State, Brazil. AB - The aim of this study was to evaluate the seroprevalence of Paracoccidioides brasiliensis infection in wild New World monkeys (Cebus sp. and Alouatta caraya). A total of 93 animals (Cebus sp., n = 68 and Alouatta caraya, n = 25) were captured in the Parana River basin, Parana State, Brazil and the serum samples were analyzed by ELISA and immunodiffusion using P. brasiliensis gp43 and exoantigen as antigens, respectively. The seropositivity observed by ELISA was 44.1% and 60% for Cebus sp. and A. caraya, respectively, while by immunodiffusion test Cebus sp. showed positivity of 2.9% only. No significant difference was observed in relation to age and sex. This is the first report of paracoccidioidomycosis in wild capuchin monkeys and in wild-black and golden howler monkeys. The high positivity to P. brasiliensis infection in both species evaluated in our study and the positivity by immunodiffusion test in Cebus sp. suggest that natural disease may be occurring in wild monkeys living in paracoccidioidomycosis endemic areas. PMID- 17914663 TI - Microenvironment-controlled encapsulation (MiCE) process: effects of PLGA concentration, flow rate, and collection method on microcapsule size and morphology. AB - PURPOSE: To evaluate the real-time effects of formulation and instrumental variables on microcapsule formation via natural jet segmentation, a new microencapsulation system termed the microenvironment-controlled encapsulation (MiCE) process was developed. METHODS: A modified flow cytometer nozzle hydrodynamically focuses an inner drug and outer polymer solution emanating from a coaxial needle assembly into a two-layer compound jet. Poly(lactic-co-glycolic acid) (PLGA) dissolved in a water-miscible organic solvent resulted in formation of reservoir-type microcapsules by interfacial phase separation induced at the boundary between the PLGA solution and aqueous sheath. RESULTS: The MiCE process produced microcapsules with mean diameters ranging from 15-25 microm. The resultant microcapsule size distribution and number of drug cores existing within each microcapsule was largely influenced by the PLGA concentration and microcapsule collection method. Higher PLGA concentrations yielded higher mean diameters of single-core microcapsules. Higher drug solution flow rates increased the core size, while higher PLGA solution flow rates increased the PLGA film thickness. CONCLUSION: The MiCE microencapsulation process allows effective monitoring and control of the instrumental parameters affecting microcapsule production. However, the microcapsule collection method in this process needs to be further optimized to obtain microcapsules with desired morphologies, precise membrane thicknesses, high encapsulation efficiencies, and tight size distributions. PMID- 17914665 TI - Researching dementia in imperial Germany: Alois Alzheimer and the economies of psychiatric practice. AB - In the writings of Alois Alzheimer and many of his contemporaries, complaints abounded about psychiatric hospitals not only failing to appreciate the importance of senile dementia, but also inhibiting scientific research into the nature and causes of the disorder. This article exploits these discontents in order to examine what Alzheimer and others thought to be optimal conditions for psychiatric research on dementia. It first analyzes the various institutional contexts in which Alzheimer worked during his career (especially in Frankfurt and Munich). It then traces some of the administrative and diagnostic practices that were deployed to enhance the conditions for his clinical and pathoanatomic research. Finally, it reflects on the implications of these practices for psychiatric care and patient experience. PMID- 17914664 TI - Pharmacokinetic-pharmacodynamic modeling of the effectiveness and safety of buprenorphine and fentanyl in rats. AB - OBJECTIVE: Respiratory depression is a serious and potentially life-threatening side-effect of opioid therapy. The objective of this investigation was to characterize the relationship between buprenorphine or fentanyl exposure and the effectiveness and safety outcome in rats. METHODS: Data on the time course of the antinociceptive and respiratory depressant effect were analyzed on the basis of population logistic regression PK-PD models using non-linear mixed effects modeling software (NONMEM). The pharmacokinetics of buprenorphine and fentanyl were described by a three- and two-compartment model, respectively. A logistic regression model (linear logit model) was used to characterize the relationship between drug exposure and the binary effectiveness and safety outcome. RESULTS: For buprenorphine, the odds ratios (OR) were 28.5 (95% CI, 6.9-50.1) and 2.10 (95% CI, 0.71-3.49) for the antinociceptive and respiratory depressant effect, respectively. For fentanyl these odds ratios were 3.03 (95% CI, 1.87-4.21) and 2.54 (95% CI, 1.26-3.82), respectively. CONCLUSION: The calculated safety index (OR(antinociception)/OR(respiratory depression)) for fentanyl of 1.20 suggests that fentanyl has a low safety margin, implicating that fentanyl needs to be titrated with caution. For buprenorphine the safety index is 13.54 suggesting that buprenorphine is a relatively safe opioid. PMID- 17914666 TI - Few preschool boys and girls with ADHD are well-adjusted during adolescence. AB - To estimate the prevalence of being well-adjusted in adolescence, boys and girls with (n = 96) and without (n = 126) attention-deficit/hyperactivity disorder (ADHD) were assessed seven times in eight years starting when they were 4-6 years of age. Symptoms of ADHD, ODD/CD, and depression/anxiety in addition to social skills and social preference were gathered using multiple methods and informants. Being well-adjusted was defined by surpassing thresholds in at least four of the five domains. At the 7- and 8-year follow-up, when youth were 11-14 years old, probands were significantly less likely to be well-adjusted relative to age- and ethnicity-matched control children. Only a minority of children with ADHD was well-adjusted in adolescence when emotional, behavioral, and social domains were considered simultaneously. Even when their ADHD symptoms improved over time, most probands exhibited significant impairment 7-8 years after their initial assessment. PMID- 17914668 TI - Invasiveness gene signature predicts a favorable outcome also in estrogen receptor-positive primary breast cancers treated with adjuvant tamoxifen. PMID- 17914667 TI - Does executive function matter for preschoolers' problem behaviors? AB - Early problem behaviors are associated with a variety of cognitive deficits: in verbal ability, executive function (EF) and theory of mind (ToM). Previous studies with different age-groups yield contrasting results: for 2-year-olds, ToM skills appear particularly salient (Hughes & Ensor, 2006), but for 4-year-olds EF appears more important (Hughes, White, Sharpen, & Dunn, 2000). To examine this apparent developmental change in the relative salience of cognitive deficits we followed 122 children from Hughes and Ensor's (2006) sample at ages 3 and 4. Tests of ToM, EF and verbal ability were included at each time-point and multi informant, multi-measure, multi-setting ratings provided aggregate measures of problem behaviors. ToM and verbal ability showed non-specific associations with problem behaviors, but associations between EF and problem behaviors were strong and specific. In addition, age-3 EF mediated relations between age-2 verbal ability and age-4 problem behaviors. PMID- 17914669 TI - Adjuvant chemotherapy for breast cancer: a cost-utility analysis of FEC-D vs. FEC 100. AB - BACKGROUND: Adjuvant 5-flurouracil, epirubicin and cyclophosphamide-docetaxel (FEC-D) has been shown to improve disease-free and overall survival (DFS and OS), compared to FEC 100, for node-positive breast cancer. An economic evaluation was undertaken to examine the cost-utility (CU) of FEC-D relative to FEC 100 given possible differences in cost between the two regimens. METHODS: A Markov model was developed to calculate the cumulative costs and quality-adjusted life years (QALY) gained over a 10-year horizon for a hypothetical cohort of 1,000 women with node-positive breast cancer treated with FEC 100 or FEC-D. Event rates, costs, and utilities were derived from the literature. Efficacy outcomes were based primarily on the hazard ratio of DFS for all patients, but separate analyses were also conducted according to age and menopausal status as per the PACS 01 subgroup analysis results. The model took a third-party direct payer perspective and reports results in 2006 Canadian dollars ($). Both costs and benefits were discounted at 3%. RESULTS: FEC-D is associated with 0.156 QALY gain and a $2,280 incremental cost compared to FEC 100, with a CU of $14,612/QALY gained. Results were robust to model assumptions and input parameters in a sensitivity analysis but were marginal in pre-menopausal and younger women. CONCLUSIONS: Adjuvant FEC-D is a cost-effective alternative to FEC 100, with a cost-effectiveness ratio well below commonly employed thresholds. The CU according to age and menopausal status should be considered in view of the potential differences in efficacy in these subgroups, if any. PMID- 17914670 TI - Efficacy of three potent steroid sulfatase inhibitors: pre-clinical investigations for their use in the treatment of hormone-dependent breast cancer. AB - Estrogenic steroids, such as estradiol, are known to play a crucial role in the development and growth of hormone-dependent breast cancer. Steroid sulfatase (STS) inhibitors that can prevent the biosynthesis of these steroids via the sulfatase pathway offer therapeutic potential. We show here the in vivo profile, including the efficacy in a xenograft breast cancer model and pharmacokinetics, of three potent STS inhibitors. MCF-7 cells stably over-expressing STS cDNA (MCF 7STS) were generated. Ovariectomised, MF-1, female nude mice receiving subcutaneous injections of estradiol sulfate (E2S) and bearing MCF-7STS xenografts, were orally treated with the STS inhibitors STX64, STX213, and STX1938. Treatment was administered once weekly at a dose of 1 mg/kg for 35 days during which animals received E2S thrice weekly. Mice were weighed and tumor measurements taken weekly. Furthermore, the pharmacokinetics for STX213 was determined in rats. STX213 and STX1938 exhibited potent STS inhibition in vivo. However, STX1938 demonstrated a greater duration of activity. In vehicle treated nude mice receiving E2S, tumor volumes increased by 260% after 35 days compared to day zero. STX64 (1 mg/kg) failed to reduce tumor growth when given once weekly. STX213 and STX1938 (once weekly, 1 mg/kg) significantly inhibited (P < 0.05) tumor growth over this same time period. These compounds completely inhibited liver and tumor STS activity and significantly reduced the levels of plasma E2. This study indicates that the STS inhibitor, STX213, exhibits excellent efficacy and pharmacokinetics and therefore offers a potentially novel treatment for hormone-dependent breast cancer. PMID- 17914672 TI - The good, the bad and the ugly: a meta-analytic review of positive and negative effects of violent video games. AB - OBJECTIVE: Video game violence has become a highly politicized issue for scientists and the general public. There is continuing concern that playing violent video games may increase the risk of aggression in players. Less often discussed is the possibility that playing violent video games may promote certain positive developments, particularly related to visuospatial cognition. The objective of the current article was to conduct a meta-analytic review of studies that examine the impact of violent video games on both aggressive behavior and visuospatial cognition in order to understand the full impact of such games. METHODS: A detailed literature search was used to identify peer-reviewed articles addressing violent video game effects. Effect sizes r (a common measure of effect size based on the correlational coefficient) were calculated for all included studies. Effect sizes were adjusted for observed publication bias. RESULTS: Results indicated that publication bias was a problem for studies of both aggressive behavior and visuospatial cognition. Once corrected for publication bias, studies of video game violence provided no support for the hypothesis that violent video game playing is associated with higher aggression. However playing violent video games remained related to higher visuospatial cognition (r (x) = 0.36). CONCLUSIONS: Results from the current analysis did not support the conclusion that violent video game playing leads to aggressive behavior. However, violent video game playing was associated with higher visuospatial cognition. It may be advisable to reframe the violent video game debate in reference to potential costs and benefits of this medium. PMID- 17914671 TI - Glycan microarrays for screening sialyltransferase specificities. AB - Here we demonstrate that glycan microarrays can be used for high-throughput acceptor specificity screening of various recombinant sialyltransferases. Cytidine-5'-monophospho-N-acetylneuraminic acid (CMP-Neu5Ac) was biotinylated at position 9 of N-acetylneuraminic acid (Neu5Ac) by chemoenzymatic synthesis generating CMP-9Biot-Neu5Ac. The activated sugar nucleotide was used as donor substrate for various mammalian sialyltranferases which transferred biotinylated sialic acids simultaneously onto glycan acceptors immobilized onto a microarray glass slide. Biotinylated glycans detected with fluorescein-streptavidin conjugate to generate a specificity profile for each enzyme both confirming previously known specificities and reveal additional specificity information. Human alpha2,6sialyltransferase-I (hST6Gal-I) also sialylates chitobiose structures (GlcNAcbeta1-4GlcNAc)(n) including N-glycans, rat alpha2,3sialyltransferase (rST3Gal-III) tolerates fucosylated acceptors such as Lewis(a), human alpha2,3sialyltransferase-IV (hST3Gal-IV) broadly sialylates oligosaccharides of types 1-4 and porcine alpha2,3sialyltransferase-I (pST3Gal-I) sialylates ganglio-oligosaccharides and core 2 O-glycans in our array system. Several of these sialyltransferases perform a substitution reaction and exchange a sialylated acceptor with a biotinylated sialic acid but are restricted to the most specific acceptor substrates. Thus, this method allows for a rapid generation of enzyme specificity information and can be used towards synthesis of new carbohydrate compounds and expand the glycan array compound library. PMID- 17914673 TI - Outcomes in a community sex offender treatment program: a comparison between polygraphed and matched non-polygraphed offenders. AB - This study compared a group of 104 adult male sex offenders who received community cognitive-behavioral treatment, correctional supervision, and periodic polygraph compliance exams with a matched group of 104 sex offenders who received the same type of treatment and supervision services but no polygraph exams. Polygraph exams focused on whether participants were following their conditions of community supervision and treatment and had avoided committing new sexual offenses. The two groups were exact pair-wise matched on three variables: (1) Static-99 risk score (Hanson & Thornton 2000, Law and Human Behavior, 24, 119 136), (2) status as a completer of prison sex offender treatment, and (3) date placed in the community. At fixed 5-year follow-up periods, the number of individuals in the polygraph group charged with committing a new non-sexual violent offense was significantly lower than in the no polygraph group (2.9% versus 11.5%). However, there were no significant between-group differences for the number of individuals charged for new sexual (5.8% versus 6.7%), any sexual or violent (8.7% versus 16.3%), or any criminal offense (39.4% versus 34.6%). The results are discussed in terms of their clinical and research implications. PMID- 17914674 TI - A lithographically-patterned, elastic multi-electrode array for surface stimulation of the spinal cord. AB - A new, scalable process for microfabrication of a silicone-based, elastic multi electrode array (MEA) is presented. The device is constructed by spinning poly(dimethylsiloxane) (PDMS) silicone elastomer onto a glass slide, depositing and patterning gold to construct wires and electrodes, spinning on a second PDMS layer, and then micropatterning the second PDMS layer to expose electrode contacts. The micropatterning of PDMS involves a custom reactive ion etch (RIE) process that preserves the underlying gold thin film. Once completed, the device can be removed from the glass slide for conformal interfacing with neural tissue. Prototype MEAs feature electrodes smaller than those known to be reported on silicone substrate (60 microm diameter exposed electrode area) and were capable of selectively stimulating the surface of the in vitro isolated spinal cord of the juvenile rat. Stretchable serpentine traces were also incorporated into the functional PDMS-based MEA, and their implementation and testing is described. PMID- 17914675 TI - Silicon-based microfilters for whole blood cell separation. AB - This paper reports on the comparison analysis of four main types of silicon-based microfilter for isolation of white blood cells (WBCs) from red blood cells (RBCs) in a given whole blood. The microfilter designs, namely, weir, pillar, crossflow, and membrane, all impose the same cut-off size of 3.5 mum to selectively trap WBCs. Using human whole blood, the microfilters have been characterized and compared for their blood handling capacity, WBCs trapping efficiency and RBCs passing efficiency. Based on the experimental results, the crossflow microfilter is superior and can be integrated with downstream components for on-chip genomic analysis. PMID- 17914677 TI - [Biological treatment of acute depressive episodes]. PMID- 17914676 TI - Adrenal causes of hypertension: pheochromocytoma and primary aldosteronism. AB - The clinical presentations of the patient with pheochromocytoma -- a rare endocrine neoplasm -- include adrenal incidentaloma, hypertensive paroxysms, sustained apparent polygenic hypertension, hypertension in pregnancy, and hypertensive crisis induced by anesthesia. Although when undiagnosed a pheochromocytoma can be lethal, it can usually be cured with surgery. Biochemical documentation with measurements of fractionated metanephrines and catecholamines should precede imaging studies. Abdomen and pelvis computed imaging is usually the first imaging test. Careful preoperative pharmacologic preparation is important for a successful surgical outcome. Adrenal pheochromocytomas can usually be removed laparoscopically, whereas, catecholamine-secreting paragangliomas typically require an open approach. All first degree relatives of pheochromocytoma patients should have biochemical testing. In addition, molecular genetic testing for germline mutations should be considered in most patients with adrenal pheochromocytoma and in all patients with paraganglioma. Primary aldosteronism is a relatively common form of secondary hypertension -- affecting 5 to 10% of all patients with hypertension. A plasma aldosterone concentration (PAC) to plasma renin activity (PRA) ratio should be obtained in patients with hypertension and hypokalemia, resistant hypertension, adrenal incidentaloma and hypertension, onset of hypertension at a young age (e.g., < 20 years of age), severe hypertension (e.g., > or =160 mm Hg systolic or > or =100 mm Hg diastolic), or whenever the clinician is considering other forms of secondary hypertension. The PAC/PRA ratio is a case finding test and a positive result should be confirmed with aldosterone suppression testing with either oral or intravenous sodium loading. The treatment goals for patients with primary aldosteronism are to prevent the morbidity and mortality associated with hypertension, hypokalemia, and cardiovascular damage. Both the subtype of primary aldosteronism and patient preference should dictate the treatment approach. PMID- 17914678 TI - Erectile dysfunction: clinical guidelines (1). AB - INTRODUCTION: According to a survey, the Massachusetts Male Aging Study, 52% of men beyond 40 years of age may have some degrees of erectile failure, and it is projected to affect 322 million men worldwide by 2025. We present a framework for the evaluation, treatment, and follow-up of the male patient who presents with erectile dysfunction. MATERIALS AND METHODS: A comprehensive review of the literature was conducted using the MEDLINE database for all articles from 1975 through 2004 on male sexual dysfunction and the most pertinent articles are discussed. RESULTS: Remarkable progress has been made in the treatment of erectile dysfunction (ED). Erectile dysfunction is a common condition associated with aging, chronic illnesses and various modifiable risk factors. Erectile dysfunction can be due to vasculogenic, neurogenic, hormonal, and/or psychogenic factors as well as alterations in the nitric oxide/cyclic guanosine monophosphate pathway or other regulatory mechanisms. The number of consultations from new patients presenting with erectile dysfunction and resulting costs for health care systems are increasing. Urologist should be the evaluating physician who supervises the surgical, medical, and hormonal treatment and who refers the patient, as necessary, to other members of the multidisciplinary team. CONCLUSION: Erectile dysfunction has a significant negative impact on quality of life. Male sexual dysfunction, especially erectile dysfunction, necessitates a comprehensive medical and psychologic evaluation involving both partners. All possible risk factors should be outlined and corrected, when feasible. PMID- 17914679 TI - Living donor kidney transplantation: how far should we go? AB - INTRODUCTION: The aim of this review was to describe the work that the Baskent University Faculty of Medicine has done to increase kidney donors' number in Turkey and also to discuss the major effects that donor-organ shortage is currently having worldwide. MATERIALS AND METHODS: From 1975 through 2003, our transplantation team at Hacettepe University Hospital and later at the Baskent University Transplantation Center (BUTC) performed 1451 kidney transplantations. Cadaver donation rates prior to and after the establishment of the National Coordination Center (NCC) were calculated and compared. Also, patient and graft survival rates for various groupings of transplantation types were compared. All statistical analysis was done using the log-rank test. RESULTS: Of all the renal transplantations completed in Turkey from 1975 to January 2004, 20% were performed by our team in our center. For the years 1990 through 2003, the 1-, 3-, and 5-year patient survival rates in the first-degree-living-related kidney transplantation group were 96%, 93%, and 91%, respectively, and the corresponding graft survival rates were 93%, 84%, and 81%. In the second-degree living-related group, the 1-, 3-, and 5-year patient survival rates were 94%, 90%, and 87%, respectively, and the corresponding graft survival rates were 93%, 86%, and 84%. For living-unrelated transplantations, the 1-, 3- and 5-year patient survival rates were 93%, 90%, and 83%, respectively, and the corresponding graft survival rates were 83%, 78%, and 76%. In the cadaver-kidney transplantation group, the 1 , 3- and 5-year patient survival rates were 85%, 78%, and 70%, respectively, and the corresponding graft survival rates were 82%, 64%, and 53%. During this same period, the 1-, 3-, and 5-year graft survival rates for our cadaver donors and living donors older than 55 years of age were 80%, 52%, 46% and 88%, 69%, 61%, respectively. CONCLUSION: Vigorous efforts by our group at Baskent University and by other transplant surgeons across the nation have increased the numbers of transplantations performed each year. As well, since the NCC was established in 2001, the number of cadaver-kidney transplantations has more than doubled. The initial results with this new nationwide organ-sharing system are promising, and there is every indication that this approach will continue to raise the number of transplant operations performed across Turkey each year. We suggest that Turkish citizens should consider changing our national policies on organ donation. Opt out policies can increase the pool of cadaver-organ transplants. In addition, to increasing cadaver donation, we feel that living-related donation restricted to first- and second-degree relatives and acceptable non-blood-related donors (such as spouses) is the best path to expanding kidney transplantation worldwide. PMID- 17914680 TI - Worldwide cadaveric organ donation systems (transplant organ procurement). PMID- 17914681 TI - A comparison between laparoscopic and open pyeloplasty in patients with ureteropelvic junction obstruction. AB - INTRODUCTION: Our aim was to compare clinical and radiological outcomes, complications, and hospital stay in laparoscopic and open pyeloplasty. MATERIALS AND METHODS: From February 2002 to February 2003, 69 patients with ureteropelvic junction obstruction (UPJO) were assigned into two groups. Thirty-seven patients underwent transperitoneal laparoscopic pyeloplasty and 32 underwent open surgical pyeloplasty. Clinical symptoms were assessed before and after surgery, subjectively. Radiological assessment was also done three months postoperatively. RESULTS: Mean operative time was 3.2 hours and 2.2 hours in laparoscopic and open pyeloplasty groups, respectively. Intraoperative bleeding was trivial in both groups and no complication or conversion to open surgery occurred. Postoperative complication rates were 24% and 6% in laparoscopic and open pyeloplasty groups, respectively. Mean hospital stay was similar (6.2 days) in the two groups. Mean follow-up was 16.5 months versus 11.4 months. Clinical and radiological success rates were 89% and 83.8% for laparoscopy group versus 96.5% and 87% for open pyeloplasty group. Due to recurrence of stricture, repeated surgery was performed in 4 patients of laparoscopy and 1 of open pyeloplasty groups. CONCLUSION: Laparoscopic pyeloplasty is a less invasive method with less pain, cosmetic advantages, no long incision, and outcome comparable with open surgery. Hospital stay is also not longer than that in open surgeries. Hence, laparoscopic pyeloplasty can be a substitute for skilled surgeons. PMID- 17914682 TI - Endoscopic renal cyst ablation. AB - INTRODUCTION: Our aim was to evaluate the result of simple renal cyst ablation by endoscope and compare the results with other techniques of renal cyst treatment. MATERIALS AND METHODS: A prospective study was performed at Shaheed Faghihi hospital from January 2001 to January 2003. Ten patients with symptomatic simple renal cyst were selected for this study. The exclusion criteria were history of previous renal surgery, parapelvic cyst, and cyst size less than 50 mm. Urinalysis, urine culture, serum electrolytes, ultrasonography, and CT scan were done before operation. The patients underwent endoscopic renal cyst ablation and cytology of cyst fluid and histopathological examination of cysts' walls were done in all patients. The patients were followed with ultrasonography after two weeks and 2, 6, and 12 months postoperatively. Disappearance of the cyst or decreasing its size to less than 50% of its primary size was considered as improvement. RESULTS: All the patients were female with a mean age of 55 (range 22 to 75) years. The operation was successful in 9 patients with no major complications. Perinephric hematoma and excessive leakage were seen in two patients. The operative time was 38+/-10.8 minutes and hospital stay was 3+/-1.3 days. Mean size of cyst before operation was 75+/-19.7 mm and changed to 12.7+/ 15.3 mm after operation (p<0.001). Flank pain subsided in 88.8% (p<0.008). CONCLUSION: Cyst ablation can be used for the treatment of simple renal cysts not responding to aspiration and sclerosing therapy, and if there is no laparoscopic facility. More studies are needed to confirm these results. PMID- 17914683 TI - PCNL in the management of lower pole caliceal calculi. AB - INTRODUCTION: Several therapeutic methods are used in the management of lower pole caliceal calculi. This survey has been conducted to evaluate the safety and efficacy of percutaneous nephrolithotomy in the management of lower pole calculi. MATERIALS AND METHODS: Fifty-five patients, 43 males and 12 females with a mean age of 41.5 (range 11 to 75) years, who had suffered from lower pole caliceal calculi and treated by standard percutaneous nephrolithotomy (PCNL) between 1997 and 2001, were enrolled in this study. The stones were classified as follows: small (less than 25 mm), intermediate (25 to 34 mm) and large (more than 35 mm). Mean follow-up was 6.2 months (range 2 weeks to 34 months). RESULTS: The stones were completely extracted by one session PCNL in 43 patients (79%). Repeat PCNL was needed in one patient and another method was used for stone extraction in another patient. Regarding the size of stone, 88%, 79%, and 74% of small, intermediate, and large stones were completely extracted, respectively. No major complication was noted. CONCLUSION: PCNL has high success rate in patients with stones larger than 2 cm and its morbidity would be low, provided that it is performed by skilled surgeons. PMID- 17914684 TI - Correlation between Apoptosis and Histological Grade of Transitional Cell Carcinoma of Urinary Bladder. AB - INTRODUCTION: To evaluate the relationship between histological grade and apoptotic index (AI) in transitional cell carcinoma (TCC) of urinary bladder. MATERIALS AND METHODS: Formalin-fixed and paraffin-embedded tissue blocks from 75 patients with TCC, who undergone transurethral resection (TUR) were studied. One 3-micron section was provided from each TUR samples. In one section after hematoxylin and eosin (H&E) staining, tumor grade was determined according to World Health Organization/International Society of Urology and Pathology (WHO/ISUP) criteria. The apoptotic cells were determined using a terminal deoxynucleotidyl transferase (TdT) mediated dUTP biotin nick end labeling (TUNEL) technique. Apoptotic index was then obtained as the percent of TUNEL positive cells from observations of at least 1000 cells in each section. RESULTS: Forty nine patients were men and 26 were women. Mean age was 56.349 years. Mean AI was 2.300.50. The relationship between grade and AI was significant (p=0.000, r=0.551); a higher grade was associated with a higher AI. CONCLUSION: Apoptosis index has a positive correlation with bladder TCC's grade. Further studies are needed to better determine the effect of apoptosis index on prognosis. PMID- 17914685 TI - Factors affecting survival in kidney recipients at kermanshah. AB - INTRODUCTION: Our aim was to evaluate patient and graft survivals in kidney recipients and factors impacting on survival rates at Kermanshah. MATERIALS AND METHODS: This study was done on 712 kidney transplants from 1989 through 2001 in Kermanshah. One of the most important applications of survival analysis is assessing the role of explanatory factors in the studied event. In this study Kaplan-Meier method was used to calculate patient and graft survivals and in order to determine the factors affecting survival, Cox proportional hazard model was used. The iterations in Cox model was four times and the inclusion and exclusion criteria, calculated by forward conditional method were less than 5% and 10%, respectively. RESULTS: Of the recipients, 47.6% were female and most of them (94.4%) had received kidneys from living unrelated donors. One-year patient survivals in recipients from living unrelated donors (LURD) and living related donors (LRD) were 89.4% and 100%, 3-year survivals were 82% and 97.4%, and 10 year survivals were 61.4% and 72%, respectively. In addition, graft survival rates in one year were 85.6% and 97.4%, in three years were 77.2% and 92.3%, and in 10 years were 33.3% and 60.6% in LURD and LRD, respectively. In Cox model, four factors, including the presence of surgical or other complications, known primary disease, and donor-recipient relationship had significant association with patient survival and seven factors, including the presence of surgical complications, known primary disease, donor-recipient relationship, gender, weight, same side transplanted kidney, and donor's age had significant relationship with graft survival. CONCLUSION: In summary, it can be concluded that patient and donor demographic characteristics and transplantation conditions may affect patient and graft survival. With the use of multivariate regression analysis methods, the characteristics that have high probability for survival can be determined. Controlling these situations, where they have high survival probability, effectively help better treatment and high survival rate. PMID- 17914686 TI - One-year efficacy of expanded polytetrafluoroethylene vascular graft in eighty three hemodialysis patients. AB - INTRODUCTION: Our aim was to evaluate the patency and efficacy of expanded Polytetrafluoroethylene (ePTFE) vascular graft in hemodialysis patients. MATERIALS AND METHODS: In a prospective study from January 1999 to January 2001 at Sina hospital 41 patients underwent implantation of 6-mm vascular grafts and 42 underwent implantation of 8-mm grafts in order to make vascular assess for hemodialysis. They were followed up to 12 months, observing the complications. RESULTS: Mean patients' age was 52.2 years. Thirty-seven of them were females and 46 were males. Over a 12 months period of follow-up, 12 cases of graft infection (14.5%), 21 cases of thrombosis (25.3%), 7 cases with both complications (8.4%), and 1 case of pseudoaneurism (1.2%) were observed. One-year patency rate was 34.9%. Diabetes was the only factor associated with lower patency rate (27% versus 57%, p<0.05). CONCLUSION: ePTFE vascular graft seems to be an appropriate substitute for arteriovenous fistula as a vascular assess in hemodialysis patients. Educating patients and good care can decrease the rate of infection, thrombosis, and other complications, resulting in a better patency and lower morbidity rate. PMID- 17914687 TI - Evaluation of urinary calculi by infrared spectroscopy. AB - INTRODUCTION: We analyzed urinary calculi composition and its relationship with gender, age, calculus weight, color, and location. MATERIALS AND METHODS: Two hundred and forty one patients with urinary calculus, who had undergone open lithotomy from June 1999 to April 2001, were enrolled in this prospective study which was performed by Tehran and Oroomieh Medical Sciences Universities. The calculi compositions were analyzed by infrared spectroscopy in Bonn University. Statistical analyses were made by paired t test. RESULTS: One hundred and forty five males with a mean age of 40.4 years and 96 females with a mean age of 42.5 years were enrolled in this study. Mean calculus weight was 4.28 gr. Mean calculus number was 4.33. Thirty four (14.1%) calculi were pure (carbonate apatite: 2, brushite: 1, uric acid: 19, cystine: 3, weddellite: 6, mono-NH4 urate: 2, struvite: 1), 207(85.6%) were mixed and none of them contained octa-ca phosphate, apatite, newberyte, 2,8-dihydroxyadenine, mono-Na-urate, or xanthine. Weddellite was found in 77% of calculi. It comprised more than 50% of them in 26% of cases. Whewellite crystals were found in 78% of calculi. It comprised more than 50% of them in 46% of cases. The most common pure calculus was uric acid and the most common component of calculi was whewellite followed by weddellite. CONCLUSION: Although there is no comprehensive study on urolithiasis incidence and prevalence in Iran, it can be concluded that whewellite and weddellite may be the most common components of urolithiasis in Iran and uric acid calculi are the most common pure calculi. There was no significant difference in calculi composition in our study. PMID- 17914688 TI - The efficacy of acupuncture in extracorporeal shock wave lithotripsy. AB - INTRODUCTION: Our aim was to evaluate the safety and efficacy of acupuncture in comparison with intravenous (IV) sedation in extracorporeal shock wave lithotripsy (ESWL). MATERIAL AND METHODS: One hundred patients, who were scheduled for ESWL, were divided randomly into two groups of fifty patients (acupuncture and IV sedation). In the first group, acupuncture was carried out with special needles (0.30 x 18 mm), in two points 30 minutes before the procedure: point of 36 from the stomach meridian with an angle of 90 degrees and point of 60 from the urinary bladder meridian with an angle of 90 degrees. In the IV sedation group morphine 0.1 mg/kg was injected intramuscularly 30 minutes and intravenous diazepam 0.1 mg/kg for muscle relaxation and anti-anxiety, one minute before the procedure. The two groups were similar in terms of confounding factors. Pain (scored in 4 levels), blood pressure, heart rate, respiratory rate, and arterial blood oxygen saturation were recorded prior to IV sedation or acupuncture, 30 minutes after each, at the beginning of ESWL, 10 minutes after ESWL, and at the end of the procedure (times 1 to 5). RESULTS: In acupuncture group the pain intensity was less than IV sedation group (for time 3, p=0.019, for time 4, p=0.002, for time 5, p=0.05). Considering the pain incidence (each pain score except zero), there was a significant difference at time 4 (p=0.012). None of the procedures was stopped because of pain and none of the patients experienced complications during operation. Arterial blood oxygen saturation was between 95% and 100% for all of the patients in acupuncture group and recovery time was faster (p<0.0001). CONCLUSION: Acupuncture is a safe and effective method for analgesia. It has a faster recovery time and economical benefits. It also provides the ability to increase the shock wave voltage. We believe that it is a good alternative for IV sedation in ESWL, particularly in patients with lung and heart disease. PMID- 17914689 TI - Treatment of renal colic using intracutaneous injection of sterile water. AB - INTRODUCTION: To evaluate the intracutaneous injection of sterile water in the treatment of renal colic. MATERIALS AND METHODS: One hundred patients with renal colic were randomly divided into two groups of 50 patients and underwent the treatment. In the first (study) group 0.5 ml of sterile water and in the control group, 0.5 ml of normal saline was intradermally injected. The severity of pain was assessed by visual analogue scale (VAS) system before and 30 and 90 minutes after the injection. Patients in whom the presence of stone was not proved were excluded from the study. RESULTS: Before the treatment mean pain severity in the study group was 9.860.4 and in the control group was 9.960.19, so that the difference was not statistically significant (p=0.12). Thirty and 90 minutes after the injection, the means were 0.762.3 and 1.022.63 in study group and 5.944 and 6.74.19 in control group, respectively. The results in 30 and 90 minutes between the two groups were statistically significant (p=0.000 and p=0.000, respectively). Pain in all patients in the study group was relieved; however, only %34 of the patients in the control group reported a decrease in pain. There was no complication among the patients of both groups and only a severe and transient pain during injection was reported by the patients. CONCLUSION: This study along with many other existing studies indicates the efficacy of intradermal injection of sterile water for the treatment of severe pain syndromes such as renal colic. The advantages of this method are its efficacy, availability, cost benefits, and easy application. We recommend the use of this approach for the treatment of renal colic. PMID- 17914690 TI - A Comparison Study between Theophylline and Placebo in Passage of Ureteral Stones. AB - INTRODUCTION: Considering the high prevalence of urinary system stones and that non-medical treatments have more costs and side effects, we decided to evaluate the effect of theophylline in the passage of ureteral stones. MATERIALS AND METHODS: One hundred and fifty patients with ureteral stones were assigned into groups A and B, whose age, sex, size of stone, and location of stone were matched together. Patients' ages ranged from 17 to 67 years. In group A theophylline (200 mg BID) was administered and group B received placebo for six weeks. Both groups were followed up by visits every fortnight and radiological assessment was performed at the end of the sixth week. The proportion of patients whose stones were passed was compared between the two groups. RESULT: In group A with theophylline consumption 46 out of 75 (61.3%) passed their stones. The mean duration between the initiation of the treatment with theophylline and stone passage was18.3 days. In group B with placebo, the stone passage occurred in 31 out of 75 (41.3%) patients (p<0.032) and the mean duration was 24.8 days (p<0.05). CONCLUSION: With regard to the findings of this study, it seems that theophylline can increase the rate of ureteral stone passage and as well, accelerate it. PMID- 17914691 TI - Endoscopic resection of lower ureter in upper urinary tract tumors. AB - INTRODUCTION: To evaluate the efficacy and safety of endoscopic resection of lower ureter in upper urinary tract tumor cases. MATERIALS AND METHODS: Five patients with transitional cell carcinoma (TCC) of the upper urinary tract were enrolled in this study. Nephrectomy was carried out through a flank incision and distal ureter with a cuff of bladder, which was removed using endoscopic approach. Complications and recurrence rate were evaluated. RESULTS: A total of 5 patients with upper urinary tract tumor underwent the endoscopic resection of lower ureter. All the patients had grade I transitional cell tumor. Two patients had suffered from bladder TCC treated previously. During the follow-up two cases developed bladder tumor: one, 18 months and another, one year postoperatively, both in the base of bladder, which was managed successfully by transurethral resection (TUR). CONCLUSION: Endoscopic resection of lower ureter in selected patients with upper urinary tract tumors can lead to lower morbidity, shorter operation time, and higher patient's satisfaction. Despite the minority of cases in this study, it seems that this method is applicable in selected cases. PMID- 17914692 TI - Adrenal myelolipoma. PMID- 17914693 TI - Large adenocarcinoma of the right adrenal cortex: a case report. PMID- 17914694 TI - Epithelioid type of paratesticular leiomyosarcoma: a case report and literature review. PMID- 17914695 TI - Renal malakoplakia simulating neoplasm in a child: successful medical management. PMID- 17914696 TI - Laparoscopic adrenalectomy: a report of the first experience in iran. PMID- 17914697 TI - Erectile dysfunction: clinical guidelines (2). AB - PURPOSE: According to a survey, the Massachusetts Male Aging Study, 52% of men beyond 40 years of age may have some degrees of erectile failure, and it is projected to affect 322 million men worldwide by 2025. We present a framework for the evaluation, treatment, and follow-up of the male patient who presents with erectile dysfunction. MATERIALS AND METHODS: A comprehensive review of the literature was conducted using the MEDLINE database for all articles from 1975 through 2004 on male sexual dysfunction and the most pertinent articles are discussed. RESULTS: Remarkable progress has been made in the treatment of erectile dysfunction (ED). Erectile dysfunction is a common condition associated with aging, chronic illnesses and various modifiable risk factors. Erectile dysfunction can be due to vasculogenic, neurogenic, hormonal, and/or psychogenic factors as well as alterations in the nitric oxide/cyclic guanosine monophosphate pathway or other regulatory mechanisms. The number of consultations from new patients presenting with erectile dysfunction and resulting costs for health care systems are increasing. Urologist should be the evaluating physician who supervises the surgical, medical, and hormonal treatment and who refers the patient, as necessary, to other members of the multidisciplinary team. CONCLUSION: Erectile dysfunction has a significant negative impact on quality of life. Male sexual dysfunction, especially erectile dysfunction, necessitates a comprehensive medical and psychologic evaluation involving both partners. All possible risk factors should be outlined and corrected, when feasible. PMID- 17914698 TI - Prostate cancer screening, yes or no? The current controversy. AB - PURPOSE: The increasing incidence of prostate cancer and different viewpoints of medical authorities to it, has lead to conversion of preliminary plan of screening test to a requisite. The objective of this study is to clarify the obscure aspects of this subject using the literature review. MATERIALS AND METHODS: We reviewed the following items in the literature: prostate cancer screening, introduction of relevant tests, screening criteria according to World Health Organization, screening experience in different countries, community notification, specialists training in order to establish an integrated approach and treatment, anxiety relief, and promotion of patient awareness in this field. RESULTS: It has been shown that, except in China, programmed and official screening of prostatic cancer has not been accepted by concordant responsible authorities, neither in developed countries nor in developing ones. However, it is performed informally in different parts of the world. CONCLUSION: There is no unanimous consensus about performance of screening for prostate cancer. Continuing voluntary referral of men above 50 years old for performing prostatic specific antigen (PSA) test has been accepted universally and is being done potentially, defined as "opportunistic screening". PMID- 17914699 TI - Correlation between prostate needle biopsy and radical prostatectomy Gleason gradings of 111 cases with prostatic adenocarcinoma. AB - PURPOSE: There are conflicting reports in the literature about correlation of biopsy and prostatectomy Gleason scores in prostate carcinoma. The goal of this study was to determine the correlation of grading in these two types of pathologic materials. MATERIALS AND METHODS: The coupled Hematoxylin and Eosin slides of 111 patients with prostate carcinoma were collected. Gleason scores were determined. Patients who had undergone any therapy except surgery were excluded from the study. Correlation between grades was calculated by determination of correlation coefficient. Accuracy of biopsy grading in prediction of final grade was also determined by measuring the sensitivity, specificity, and positive and negative predictive values. RESULTS: In 50 cases (45%), grade was underestimated in the biopsy. After dividing the cases into Gleason scores of 2 to 4, 5 to 6, 7, and 8 to 10, the most of undergraded cases (84.2%) were in the first group (Gleason score 2 to 4) and this rate reached 5% in the fourth group (Gleason score 8 to 10). The correlation coefficient measured was 0.535 in grade to grade comparing and 0.514 in group to group comparison of the specimens. In low-grade tumors, grading in biopsy, in spite of high sensitivity (90.9%), had low positive predictive value (26.3%). CONCLUSION: There is a moderate direct linear relationship between scores in biopsy and prostatectomy specimens. But there is a high probability of underestimation of real Gleason score of the radical prostatectomy specimen in low-grade tumors. Pathologists and urologists must consider the phenomenon of undergrading in reporting prostate specimens and managing patients. PMID- 17914700 TI - Effect of smoking on prognostic factors of transitional cell carcinoma of the bladder. AB - PURPOSE: This study was conducted to evaluate the effects of smoking on the clinical characteristics and growth trend of transitional cell carcinoma (TCC) of the bladder. MATERIALS AND METHODS: In a retrospective case-control study from February 2000 to March 2003, patients with TCC of bladder, referred to our clinic, were selected and divided into high-grade and low-grade groups. Groups were matched for other known risk factors and the effect of smoking on size, number, and presenting grade of TCC in each group was evaluated. RESULTS: A total of 185 patients, with a mean age of 65.1 +/- 14.0 year, were included in this study, of whom 36 were females and 149 were males (male to female ratio of 4.1 to 1). Eighty-three patients were smokers (44.9%) with a mean 20.01 +/- 11.09 pack year (range 0.75 to 60) smoking history. History of smoking was positive in 36.1% of the patients with low-grade tumors; whereas, 90% of the patients with high grade tumors were smokers (P = 0.000, OR = 15.9, 95% CI: 6.7-36.9). There was a statistically significant correlation between the history of smoking and size and number of tumoral lesions (P = 0.000, P = 0.000, respectively). Positive history of smoking was also associated with higher grades of tumor in both men and women (OR = 12.8 and 8.8, respectively). CONCLUSION: This study showed that smoking not only induces bladder cancer, but also, once it develops, it can increase the grade of tumor, resulting in worse prognosis. Thus, smoking cessation might favorably alter the course of bladder cancer. PMID- 17914701 TI - Is bladder cancer more common among opium addicts? AB - PURPOSE: Many environmental and occupational risk factors have been proposed for bladder cancer, among which opium consumption has been considered in few studies. We designed a study to determine the relationship between opium consumption and bladder cancer. MATERIALS AND METHODS: In a retrospective, case-control study, male patients with bladder cancer, who had been referred to our hospital in a three-year period, were selected. Data regarding age, gender, smoking, and opium consumption were collected from patients' records and compared with data of a control group, consisting of patients with benign prostatic hyperplasia (BPH). RESULTS: Fifty-two male patients with bladder tumor (group 1) were compared with 108 patients with BPH (group 2). Of the patients with bladder cancer, 36 (68%) were smokers, of whom 12 were also opium addicts. In general, 13 (25.5%) patients were opium consumers (one opium consumer was not smoker). From 108 patients with BPH, 25 (23%) were smokers, of whom, 5 were also opium addicts. Mean duration of cigarette smoking was 31 +/- 13.6 and 20.2 +/- 14.7 years in patients with bladder cancer and BPH, respectively. The duration of opium consumption was 11.9 +/- 1.4 and 6.2 +/- 1.3 years in groups 1 and 2, respectively. The duration of cigarette smoking and opium consumption in group 1 was greater than that in group 2. In addition, smoking increases the risk of bladder cancer 3.8-fold (OR = 8.3, 95% CI = 1.8-7.8). Simultaneous cigarette smoking and opium consumption increases the risk of bladder cancer 6.2-fold (OR = 6.2, 95% CI = 2.04-18.7). CONCLUSION: There are few studies regarding the carcinogenic effect of opium on bladder. We demonstrated that, the incidence of bladder cancer in smokers, who are simultaneously opium consumers, was higher than in patients who were only smokers. Simultaneous opium addiction and cigarette smoking may have some roles in the pathogenesis of bladder tumor. However, further studies with large sample sizes are warranted. PMID- 17914702 TI - A comparative study on the effect of lidocaine and furosemide on urinary output and graft function after renal transplantation. AB - PURPOSE: Renal transplantation is an ideal treatment for patients with chronic renal failure. It was demonstrated that despite the adhesion to surgical and anesthetic principles, urinary output is not satisfactory after transplantation. It seems that microvascular spasm of renal vasculature is responsible for this phenomenon. We designed a study to investigate whether lidocaine injection into renal artery can relieve vasospasm and subsequently improve output and graft function better than furosemide. MATERIALS AND METHODS: In a randomized clinical trial, from July 2002 to November 2003, 100 consecutive patients who were referred to our center for kidney transplantation were recruited in this study. After obtaining written informed consent, they were divided blindly into two groups. In group 1, lidocaine was injected into renal artery, before arterial anastomosis, and group 2 received furosemide as the conventional intervention. Urine volume within 1, 4, and 24 postoperative hours and serum creatinine levels in the first three weeks were recorded and compared between the two groups. RESULTS: Urine volumes at 1, 4, and 24 hours after transplantation were higher significantly in lidocaine group (P <0.001). Serum creatinine levels were lower significantly in the first postoperative day and also 21 days after transplantation in group 1 (P <0.001). CONCLUSION: Comparing to furosemide, it seems that lidocaine can cause a more effective vasodilation in renal arteries of kidney allograft, resulting in a better diuresis. This may have a role in the betterment of graft function. PMID- 17914703 TI - Incisional hernia after renal transplantation and its repair with propylene mesh. AB - PURPOSE: Kidney recipients are susceptible to incisional hernia. We studied predisposing factors for incisional hernia in our patients and the results of its repair with propylene mesh. MATERIALS AND METHODS: From 1989 to 2002, 589 patients had undergone kidney transplantation in our hospital. Of these, patients who developed incisional hernia were evaluated in this study. The following data were collected from their records: age, gender, weight, age at graft rejection, surgical complications, treatment method, and the treatment results with propylene mesh. RESULTS: Of 589 recipients, 16 (3%) developed incisional hernia in surgical site. The median interval between kidney transplantation and developing of incisional hernia was 48 (range 12 to 425) days. Predisposing factors were overweight, age over fifty years, and female gender (P <0.005). In four patients, hernia was small, and the repair was performed without using mesh. Three patients were reluctant to hernia repair, and in 9, due to the large size of hernia, repair was done using propylene mesh. Having these 9 patients treated with propylene mesh, 2 developed serous collection in surgical site, which were managed successfully with multiple punctures. Hernia recurrence or infection was not noted in these patients during 3 to 36 months follow-up periods. CONCLUSION: Incisional hernia is not a rare entity after kidney transplantation. Predisposing factors, such as overweight, age over 50 years, and female gender have a role in its development. Also, repeated surgeries in kidney recipients can increase the risk of incisional hernia. Managing this complication with propylene mesh is a safe and effective method. PMID- 17914704 TI - The results of grade IV cystocele repair using mesh. AB - PURPOSE: To evaluate the results of grade IV cystocele repair by 4-corner bladder and bladder neck suspension technique, using prolene mesh. MATERIAL AND METHODS: Thirty-one women with a median age of 61 years and severe anterior vaginal wall prolapse (grade IV cystocele) were treated by 4-corner bladder and bladder neck suspension technique, using prolene mesh. Of these, 3 had associated uterine prolapse, rectocele, and enterocele, one had rectocele and enterocele, and 18 had rectocele only. In these cases, pelvic floor defects were also repaired simultaneously and in 3, vaginal hysterectomy was done. Twelve patients had a previous failed cystocele repair. In a 32-month follow-up, the patients were evaluated with vaginal examination and upright cystography. Urinary continence during increased intra-abdominal pressure was also assessed, based on subjective symptoms. RESULTS: None of the patients had cystocele recurrence. Urinary continence during increased intra-abdominal pressure was seen in all of the patients. Intraoperative rectal or bladder injury did not occur. Transfusion was not required in any of the cases. Early complications (6 to 8 weeks postoperatively) included irritative urinary symptom in 17 patients, of whom, 8 had documented urinary tract infection that were treated successfully. Late complications were spotting in 3 cases (two were treated with topical estrogen and vaginal mucosal repair was done in one), dyspareunia in 4 sexually active patients, changes in urination pattern in 28 (improved significantly with behavioral therapy), long-term urge incontinence (>8 weeks) in 5 (medical treatment was successful in these patients), and prolonged intermittent catheterization in 1. Pelvic abscess and migration of mesh were not observed. CONCLUSION: According to our findings, using mesh in patients with grade IV cystocele, who had a previous failed surgery or weakness in supportive pelvic tissue, is an appropriate treatment modality. PMID- 17914705 TI - The effect of camphor on the male mice reproductive system. AB - PURPOSE: In Iranian traditional medicine there is a belief that camphor is a suppressor of sexual activity. Based on this idea and since there are few studies on this issue, we evaluated the effect of camphor on histopathological changes of reproductive system in young male mice of balb/c racial type. MATERIALS AND METHODS: Thirty-six premature male balb/c mice, were divided into 3 paired groups of experimental, control, and sham (n = 6). Experimental groups 1 and 2 received 30 mg/kg camphor dissolved in olive oil (orally) for 10 and 20 days, respectively. The control groups received the same volume of olive oil during the same periods of time, and no intervention was done in sham groups. All groups were kept in the same environmental condition. At the end of exposure time, each group was anesthetized and their testes were removed for obtaining serial sections, and histological staining. RESULTS: Comparing to the control groups less vascularization in testis tissue of experimental groups was seen. Furthermore, using stereological methods demonstrated that internal diameters of seminiferous tubules in experimental groups were significantly smaller than those in control groups (P <0.005). Also, the number of released sexual cells was lower in experimental groups (P <0.005). No meaningful difference was seen between controls and sham groups. CONCLUSION: Administration of camphor and its effects on male mice reproductive system may result in significant structural changes, including vascularization and proliferation of sexual cells. This can affect maturation of seminiferous tubules and subsequently, reproductive function of testes in mice. PMID- 17914706 TI - The correlation between semen parameters in processed and unprocessed semen with pregnancy rate in intrauterine insemination in the treatment of male factor infertility. AB - PURPOSE: Our aims were to determine the relation of semen parameters in processed and unprocessed semen samples with pregnancy rate in intrauterine insemination (IUI) in the treatment of male factor infertility. MATERIALS AND METHODS: In a quasi experimental study, 412 couples with male factor infertility were studied. To treat male factor infertility, 561 IUI cycles were done. RESULTS: Total pregnancy rate was 7.84% (44 in 561) per cycle. There was an inverse relationship between pregnancy rate and duration of infertility. Total sperm count after processing was higher in pregnant cycles than in non-pregnant ones (P <0.05). The mean total motile sperms after processing for pregnant and non-pregnant cycles was 72.2 +/- 78.8 and 53.2 +/- 54.3, respectively (P <0.05). CONCLUSION: IUI is a valuable method for the treatment of male factor infertility. The higher number of sperms, total motile sperms and IUI sessions, and lower duration of infertility, all have a positive relationship with pregnancy rate. PMID- 17914707 TI - Huge benign prostatic hyperplasia. PMID- 17914709 TI - Pseudoaneurysm following percutaneous nephrolithotomy. PMID- 17914708 TI - Primary tuberculosis of glans penis: a case report. PMID- 17914710 TI - Successful medical treatment of emphysematous pyelonephritis. PMID- 17914711 TI - Re: treatment of renal colic using intracutaneous injection of sterile water. PMID- 17914712 TI - Preservation of effect and the regulatory approval of new treatments on the basis of non-inferiority trials. AB - The criteria for approval of an experimental treatment on the basis of active controlled non-inferiority trials often include demonstration of 'preservation of effect' relative to the active control. While this appears on its surface to be a reasonable criterion, on closer inspection it can be shown to lead to serious logical inconsistencies. In particular, an experimental treatment may have clinical trial results that are superior to those for the standard treatment and yet fail to meet this criterion for approval. In this paper we propose a set of principles that will help avoid these logical inconsistencies, and we argue that the qualities of an experimental treatment that are required for approval should be consistent regardless of the presence or absence of existing treatments and the type of study design used to evaluate the treatment. PMID- 17914713 TI - Misspecification tests for binomial and beta-binomial models. AB - The IOS test of Presnell and Boss (J. Am. Stat. Assoc. 2004; 99(465):216-227) is a general-purpose goodness-of-fit test based on the ratio of in-sample and out-of sample likelihoods. For large samples, the IOS statistic can be approximated by a multiplicative contrast between two estimates of the information matrix, and in this way the IOS test is connected to White's (Econometrica 1982; 50:1-26) information matrix test, or IM test, which is based directly on the difference of two estimates of the information matrix. In this paper, we compare the performance of IOS to that of the IM test and of other goodness-of-fit tests for binomial and beta-binomial models, in both examples and simulations. Our findings suggest that IOS is strongly competitive, not only against the IM test but also against tests designed for specific binomial and beta-binomial models. PMID- 17914714 TI - Sequential causal inference: application to randomized trials of adaptive treatment strategies. AB - Clinical trials that randomize subjects to decision algorithms, which adapt treatments over time according to individual response, have gained considerable interest as investigators seek designs that directly inform clinical decision making. We consider designs in which subjects are randomized sequentially at decision points, among adaptive treatment options under evaluation. We present a sequential method to estimate the comparative effects of the randomized adaptive treatments, which are formalized as adaptive treatment strategies. Our causal estimators are derived using Bayesian predictive inference. We use analytical and empirical calculations to compare the predictive estimators to (i) the 'standard' approach that allocates the sequentially obtained data to separate strategy specific groups as would arise from randomizing subjects at baseline; (ii) the semi-parametric approach of marginal mean models that, under appropriate experimental conditions, provides the same sequential estimator of causal differences as the proposed approach. Simulation studies demonstrate that sequential causal inference offers substantial efficiency gains over the standard approach to comparing treatments, because the predictive estimators can take advantage of the monotone structure of shared data among adaptive strategies. We further demonstrate that the semi-parametric asymptotic variances, which are marginal 'one-step' estimators, may exhibit significant bias, in contrast to the predictive variances. We show that the conditions under which the sequential method is attractive relative to the other two approaches are those most likely to occur in real studies. PMID- 17914715 TI - Controlling the type 1 error rate in non-inferiority trials. AB - Two different approaches have been proposed for establishing the efficacy of an experimental therapy through a non-inferiority trial: The fixed-margin approach involves first defining a non-inferiority margin and then demonstrating that the experimental therapy is not worse than the control by more than this amount, and the synthesis approach involves combining the data from the non-inferiority trial with the data from historical trials evaluating the effect of the control. In this paper, we introduce a unified approach that has both these approaches as special cases and show how the parameters of this approach can be selected to control the unconditional type 1 error rate in the presence of departures from the assumptions of assay sensitivity and constancy. It is shown that the fixed margin approach can be extremely inefficient and that it is always possible to achieve equivalent control of the unconditional type 1 error rate, with higher power, by using an appropriately chosen synthesis method. PMID- 17914716 TI - Skin peptides: biological activity and therapeutic opportunities. AB - The skin provides an effective barrier to the loss of body fluids and environmental assault. In addition to the physical barrier provided by the stratum corneum, the skin also contains a chemical barrier consisting of antimicrobial peptides (AMPs), which control microbial growth on the surface. These AMPs also have multiple roles as mediators of inflammation with effects on epithelial and inflammatory cells, influencing cell proliferation, wound healing, cytokine/chemokine production and chemotaxis. This review describes the range of peptides found in the skin, both constitutive and those induced in response to injury. The role these peptides play in normal skin function and in various skin conditions is described. A better understanding of their role in normal and skin disease may offer new strategies in skin disease, dermatology and as cosmeceuticals. PMID- 17914717 TI - Weisses Blut--Leukapheresis in acute leukemia. PMID- 17914718 TI - N-in-1 dosing pharmacokinetics in drug discovery: experience, theoretical and practical considerations. AB - N-in-1 (or cassette) dosing pharmacokinetics (PK) has been used in drug discovery for rapid assessment of PK properties of new chemical entities. However, because of potential for drug-drug interactions this procedure is still controversial. This study was to retrospectively evaluate the N-in-1 dosing approach in drug discovery with an emphasis on the potential for drug-drug interactions. The systemic clearance, volume of distribution, oral bioavailability, and renal excretion of the 31 lead compounds in rats, dogs or chimpanzees were significantly correlated between the N-in-1 dosing and discrete studies with r values of 0.69, 0.91, 0.53, and 0.83 (p < 0.005 for all), respectively. PK parameters for 11 quality control compounds which were involved in 194 N-in-1 studies for screening approximately 1000 compounds had coefficient of variations of less than 70%. The intrinsic microsomal clearances generated from the N-in-1 and discrete incubations were nearly identical (r = 0.97, p < 0.0001). The intrinsic clearances of quality control compound from the N-in-1 incubations were consistent with its discrete CL(int) estimate (cv: 5.4%). Therefore, N-in-1 dosing is a useful approach in drug discovery to quickly obtain initial PK estimates. Potential drug-drug interactions that result in confounding PK estimates do not occur as frequently as expected. PMID- 17914719 TI - Contemporary encephalitis lethargica presenting with agitated catatonia, stereotypy, and dystonia-parkinsonism. AB - Encephalitis lethargica (EL) syndrome was classically described by Von Economo and has somnolent-ophthalmoplegic, hyperkinetic, and amyostatic-akinetic forms. We describe 2 recent cases of EL characterized by an acute encephalitis with mixed movement disorders (dystonia-Parkinsonism plus stereotypy) and psychiatric disorders (agitated catatonia, coprolalia, and echo phenomena). Both patients suffered concurrent hyperkinetic and Parkinsonian features resulting in therapeutic challenges. Bradykinetic features responded to dopamine replacement therapy and both patients also had adverse affects to dopamine antagonists (oculogyric crises plus neuroleptic malignant syndrome). Investigation was unremarkable other than the presence of CSF lymphocytosis and oligoclonal bands. Despite prolonged in-patient stays and intensive care management, both patients have made complete recoveries. We believe these cases support the hypothesis that this syndrome is an inflammatory encephalitis that specifically effects dopamine neurotransmission. PMID- 17914720 TI - Paradoxical kinesia at war. AB - Paradoxical kinesia is the sudden transient ability of a patient with Parkinson's disease to perform a task he was previously unable to perform, usually when facing an immediate threat. The sensory cues governing this behavior and the prevalence in real life situations are unknown. The objective of this study was to determine the occurrence of paradoxical kinesia in Parkinson's disease (PD) patients whose residential area was suddenly a war zone, under a life threatening missile attack, necessitating immediate evacuation. Fifty PD patients were interviewed during and immediately following the war. Only two patients experienced paradoxical kinesia, one war related and the other historical, both in response to visual cues. In contrast, an auditory stimulus in the form of a frightening loud siren, warning patients of an imminent missile attack, did not induce paradoxical kinesia. When questioned about their general function during wartime, patients reported significant increases in OFF time (P < 0.01), dyskinesia (P < 0.009), anxiety (P < 0.002), and depression (P < 0.01) as compared with their performance before the war. Paradoxical kinesia is uncommon, even in the face of danger. Visual, but not auditory, triggers appear to be needed to prompt its occurrence. PMID- 17914721 TI - Reversible dopamine agonist induced anterocollis in a multiple system atrophy patient. PMID- 17914722 TI - Accuracy of DaTSCAN (123I-Ioflupane) SPECT in diagnosis of patients with clinically uncertain parkinsonism: 2-year follow-up of an open-label study. AB - We previously reported on the role of dopamine transporter (DAT) SPECT in the workup of patients with clinically uncertain parkinsonian syndromes (CUPS). The findings of that study supported the use of SPECT imaging with DaTSCAN ((123)I Ioflupane) for accurate diagnosis in this population. We report here the 2-year follow-up of the CUPS study, which aimed to validate the results of DaTSCAN imaging and to ascertain whether a second scan could minimize any residual diagnostic uncertainty among those with an inconclusive diagnosis. Eighty-five of 118 patients (72%) were available at follow-up. In 8 of 85 patients the neurologist was unable to provide a definite diagnosis (named as inconclusive). At follow-up, clinical diagnosis agreed with initial DaTSCAN SPECT results in 69 of 77 patients (90%) in whom a specific diagnosis was established. A second SPECT scan was performed if clinical diagnosis at follow-up differed to that suggested by the initial scan (n = 8) or was inconclusive (n = 8). Among 8 patients whose clinical diagnosis differed to DaTSCAN images, a second scan was performed in 6 (2 refused) and the results supported the final clinical diagnosis in 4. Follow up DaTSCAN SPECT helped to establish a diagnosis in 7 of 8 patients (87.5%) with a previously inconclusive diagnosis. DaTSCAN imaging shows a high rate of agreement with clinical diagnosis after 2-years follow-up. A second scan at 2 years follow-up can reduce remaining diagnostic uncertainty that is present even after a prolonged period of observation. PMID- 17914723 TI - Cooling can relieve the difficulty of playing the tuba in a patient with embouchure dystonia. PMID- 17914724 TI - Successful treatment of tremor in Wilson's disease by thalamotomy: A case report. AB - Little information is available on the surgical treatment of movement disorders in Wilson's disease. We report a successful outcome of left-sided stereotactic thalamotomy in a 30-year-old man with Wilson's disease, who had severe postural kinetic tremor of both hands. The improvement was bilateral. Our case illustrates that stereotactic thalamotomy may be considered as an option in treating severe tremor in selected patients of Wilson's disease and merit further trials. PMID- 17914725 TI - A unique case of coincidence of early onset Parkinson's disease and multiple sclerosis. AB - We report on a patient who developed left arm rest/postural tremor at age 24 and responded well to trihexyphenidyl. One year later spastic paraparesis appeared, and multiple sclerosis was diagnosed on the basis of clinical, radiological, and laboratory evidence. Although paraparesis improved after immunosuppressant therapy, a complete picture of an asymmetric parkinsonian syndrome gradually developed. Excellent response to levodopa, drug-induced dyskinesias, and DaTSCAN revealing pathology congruent with Parkinson's disease (PD) indicate a coincidental etiopathogenetic relationship of both clinical entities: multiple sclerosis and PD. Genetic analyses focusing on autosomal recessive parkinsonism (parkin, DJ1, and PINK1) were negative. To the best of our knowledge, only 15 cases of parkinsonism in association with multiple sclerosis have been reported, and their relationship has been interpreted to be either causal or coincidental. This is the first report of a coincidence of both entities, in which the parkinsonian syndrome developed first and before age 30. PMID- 17914726 TI - Multiplex ligation-dependent probe amplification assay for simultaneous detection of Parkinson's disease gene rearrangements. AB - Parkinson's disease (PD) is a common disorder caused by degeneration of dopaminergic neurons in the substantia nigra and other brain areas. Mutations in several genes have been associated with both autosomal dominant PD and recessive early onset Parkinsonism (EOP). Genomic rearrangements such as deletions or multiplications of one or more exons represent a common mutational mechanism for most of these genes and are not detectable with routine mutation screening techniques. MLPA (Multiplex Ligation-dependent Probe Amplification), is a cheap, simple, rapid, and sensitive tool to detect exon dosage alterations and specific point mutations in selected genes. We tested the recently developed PD-MLPA assay by using 13 positive control samples carrying known mutations in SNCA, LRRK2, Parkin, PINK1, and DJ-1 genes. We then applied this technique to screen 16 EOP patients who were then cross-tested by quantitative PCR (qPCR). All the mutations present in the positive control samples were clearly detected by MLPA. Moreover, three novel Parkin rearrangements were identified among EOP patients and confirmed by qPCR. Only two samples generated false positive duplications of LRRK2 exon 1 and UCH-L1 exon 9, respectively. These results show that PD-MLPA assay can simultaneously and effectively detect rearrangements in most PD genes (SNCA, Parkin, PINK1, and DJ-1) as well as the LRRK2 G2019S common mutation. Thus, the use of this novel platform can improve the analysis of such mutations, facilitating comprehensive genetic testing in PD and EOP. PMID- 17914727 TI - Potential outcome measures and trial design issues for multiple system atrophy. AB - Multiple system atrophy (MSA) is a neurodegenerative disorder exhibiting a combination of parkinsonism, cerebellar ataxia, and autonomic failure. A disease specific scale, the Unified Multiple System Atrophy Rating Scale (UMSARS), has been developed and validated to measure progression of MSA, but its use as an outcome measure for therapeutic trials has not been evaluated. On the basis of twelve months of follow-up from an observational study of 67 patients with probable MSA, we evaluated three disease-specific scores: Activities of Daily Living, Motor Examination, and a combined score from the UMSARS and two general health scores, the Physical Health and Mental Health scores of the SF-36 health survey, for their use as outcome measures in a therapeutic trial. We discuss related design issues and provide sample size estimates. Scores based on the disease-specific UMSARS seemed to be equal or superior to scores based on the SF 36 health survey. They appeared to capture disease progression, were well correlated and required the smallest sample size. The UMSARS Motor Examination score exhibited the most favorable characteristics as an outcome measure for a therapeutic trial in MSA with 1 year of follow-up. PMID- 17914728 TI - Abnormal movements in Rett syndrome are present before the regression period: a case study. AB - The suspicion of a diagnosis of Rett syndrome (RTT) is based on clinical criteria that are often not present in the first two stages of the disease, as many of its symptoms will appear at a later age. This sometimes postpones the genetic diagnosis and an early clinical intervention. We present the case of 19-months old girl who came to the consultation because of an arrest of psychomotor development noticed 5 months earlier without change in sleep pattern, behavior, or social communication. In the observation of 1 hour videotape, she presented subtle stereotypic movements of the face and hands as well as repetitive dystonic posturing of her limbs. A genetic test confirmed the diagnosis of RTT, showing a truncating mutation in the MECP2 gene (R270X). This case confirms that stereotypic movement anomalies, albeit infrequent and subtle, are already present before the regression stage and while maintaining prehension and that, in addition, repetitive dystonic postures may occur. Recognition of these early movement disorders will improve clinicians' ability to perform an earlier diagnosis of RTT. PMID- 17914729 TI - Predictors of punding in Parkinson's disease: results from a questionnaire survey. AB - Dopamine replacement therapy (DRT) for Parkinson's disease (PD) has recently been linked to the development of a number of nonmotor behavioral control problems. Punding, one of these nonmotor problems, is a term used to describe complex, purposeless stereotyped behaviors such as the repetitive handling or sorting of objects. A self-report questionnaire was adapted to assess punding in the context of dysfunctional hobby-related activities. We report the results of a survey of PD outpatients from a PD research clinic (n = 141) and non-PD controls (n = 103); conducted to identify clinical and psychological factors predictive of punding behaviors. The PD group reported hobbies and activities, which scored significantly higher on the Punding Scale than controls. Higher impulsivity, poorer disease-related quality of life, younger age of disease onset, and concomitant daily medication dosage from dopamine receptor agonists were independently predictive of higher Punding Scale scores in the PD group. These findings are similar to those seen in dopamine dysregulation syndrome, and provide further evidence for the role of impulsivity and age at disease onset in DRT-related nonmotor behavioral problems in PD. PMID- 17914730 TI - Measuring health-related quality of life in MSA: the MSA-QoL. AB - The objective of this study was to develop a new patient-reported outcome measure for patients with multiple system atrophy (MSA) and to test its psychometric properties. There were three stages. First, a pool of potential scale items was generated from in-depth patient interviews. Second, these items were administered, in the form of a questionnaire, to a sample of people with MSA and traditional psychometric methods used to develop a rating scale satisfying standard criteria for reliability and validity. Third, the psychometric properties of the rating scale were examined in a second sample. In stage one, a pool of 105 items was generated from 20 patient interviews. In stage two, a scale with three subscales (motor, 14 items; nonmotor, 12 items; emotional/social functioning, 14 items), satisfying standard criteria for reliability and validity, was developed from the response data of 317 patients with MSA (response rate 71%). In stage three, the scale was examined in 286 people with MSA. Missing data were low, scores in both subscales were evenly distributed, and floor and ceiling effects were small. Reliability was high (Cronbach's alpha 0.83-0.93; test-retest ICC 0.88-0.92). Validity was supported by the interscale correlations (r = 0.47-0.59), known group differences, and the magnitude and pattern of correlations with four other rating scales, disease severity, and disease duration. In conclusion, the patient-rated MSA health-related Quality of life scale (MSA-QoL) may be a suitable patient-reported scale for use in clinical trials and studies in MSA. PMID- 17914731 TI - Evaluation of hydrate-screening methods. AB - The purpose of this work is to evaluate the effectiveness and reliability of several common hydrate-screening techniques, and to provide guidelines for designing hydrate-screening programs for new drug candidates. Ten hydrate-forming compounds were selected as model compounds and six hydrate-screening approaches were applied to these compounds in an effort to generate their hydrate forms. The results prove that no screening approach is universally effective in finding hydrates for small organic compounds. Rather, a combination of different methods should be used to improve screening reliability. Among the approaches tested, the dynamic water vapor sorption/desorption isotherm (DVI) method and storage under high humidity (HH) yielded 60-70% success ratios, the lowest among all techniques studied. The risk of false negatives arises in particular for nonhygroscopic compounds. On the other hand, both slurry in water (Slurry) and temperature cycling of aqueous suspension (TCS) showed high success rates (90%) with some exceptions. The mixed solvent systems (MSS) procedure also achieved high success rates (90%), and was found to be more suitable for water-insoluble compounds. For water-soluble compounds, MSS may not be the best approach because recrystallization is difficult in solutions with high water activity. Finally, vapor diffusion (VD) yielded a reasonably high success ratio in finding hydrates (80%). However, this method suffers from experimental difficulty and unreliable results for either highly water-soluble or water-insoluble compounds. This study indicates that a reliable hydrate-screening strategy should take into consideration the solubility and hygroscopicity of the compounds studied. A combination of the Slurry or TCS method with the MSS procedure could provide a screening strategy with reasonable reliability. PMID- 17914732 TI - Conformation and side chains environments of recombinant human interleukin-1 receptor antagonist (rh-IL-1ra) probed by raman, raman optical activity, and UV resonance Raman spectroscopy. AB - The conformation and local environments of the side chains cysteines and aromatics of recombinant human interleukin-1 receptor antagonist (rh-IL-1ra) have been studied by visible Raman, Raman optical activity (ROA) and UVRR spectroscopy. The results reveal that the secondary structure of rh-IL-1ra is predominantly beta-sheet, which is consistent with conclusions from multinuclear NMR in solutions and X-ray diffraction analysis of crystals. It confirms that all four cysteines are in reduced state. Three cysteines are not hydrogen bonded and exposed. One cysteine is moderately hydrogen bonded and buried. This explains the earlier observation that only three cysteines were detectable using DTNB titration. No characteristic Raman band of disulfide bond was observed in the Raman spectra of rh-IL-1ra in both solution and in crystals. It rules out the supposition that there is one disulfide bond in rh-IL-1ra crystals based on X-ray diffraction. Raman and UVRR spectra of rh-IL-1ra exhibit canonical marker bands of tryptophan. They do not support the proposal that there is cation-pi interaction involving tryptophans in solutions and crystals. These results demonstrate that Raman spectroscopy offers certain advantages over X-ray diffraction for studies of detailed local environment and intermolecular interactions of side chains of proteins in solution and in crystals. PMID- 17914733 TI - Initial testing of dasatinib by the pediatric preclinical testing program. AB - BACKGROUND: Dasatinib, a dual inhibitor of the src and abl tyrosine kinases, was recently approved by the Federal Drug Administration for the treatment of imatinib mesylate-resistant chronic myeloid leukemia. PROCEDURES: Dasatinib was tested against the Pediatric Preclinical Testing Program (PPTP) in vitro panel at concentrations ranging from 0.1 nM to 1.0 microM and was tested in vivo at a dose of 50 mg/kg administered orally twice daily 5 days per week for 4 weeks for the solid tumor xenografts and once daily for the acute lymphoblastic leukemia (ALL) xenografts. RESULTS: Dasatinib was selectively active against the cell lines of the PPTP in vitro panel, reaching an IC(50) in 6 of the 22 lines. The most sensitive were the AML line Kasumi-1, which has a gain-of-function c-Kit mutation (Asn822Lys), and the rhabdoid tumor line CHLA-266 (IC(50) approximately 10 nM for each). In the in vivo panel, dasatinib induced significant differences in EFS distribution in 8 of 32 (25%) solid tumor models and 3 of 7 ALL models. Using the time to event activity measure, dasatinib had intermediate activity against 1 of 27 (4%) evaluable solid tumor xenografts and 3 of 7 ALL xenografts. One xenograft in the ALL panel, a Philadelphia chromosome positive (Ph(+)) ALL xenograft, demonstrated a complete response. CONCLUSIONS: Dasatinib was active at low nanomolar concentrations against a small subset of the PPTP's in vitro panel. Dasatinib had limited in vivo activity against the PPTP solid tumor xenografts, but was highly active against a Ph(+) ALL xenograft and also had anti-leukemia activity against two other xenografts. PMID- 17914735 TI - Review of image defined risk factors in localized neuroblastoma patients: Results of the GPOH NB97 trial. AB - BACKGROUND: Recently, an international expert group proposed revision of the International Neuroblastoma Staging System (INSS). Localized disease can be classified as L1 without and as L2 with image defined risk factors (IDRF published in JCO 2005; 23:8483-8489). Our aim was to evaluate IDRF for the prediction of resectability, complications, and outcome. PROCEDURE: Records of 520 localized neuroblastoma patients of the NB97 trial were reviewed. Patients were retrospectively classified as having IDRF or not. A total of 366 evaluable patients were then analyzed for extent and complications of surgery and the prognostic value of IDRF. RESULTS: Any IDRF was present in 26/160 of stage 1, 49/113 of stage 2, and 64/93 of stage 3 patients. Complete primary resection was achieved in 156/227 patients without IDRF and 43/139 patients with IDRF (P < 0.001). The frequency of complications was higher if any IDRF was present: 37/139 versus 33/227 (P = 0.006). Lack of IDRF was associated with better event free survival (3-year-EFS 86 +/- 2% vs. 75 +/- 4%, P = 0.010), whereas overall survival was similar (3-year-OS 98 +/- 1% vs. 96 +/- 2%, P = 0.462). EFS clearly depended on INSS stage (3-year-EFS 93 +/- 2% in stage 1, 78 +/- 4% in stage 2, and 69 +/- 5% in stage 3, P < 0.001). OS was not different (3-year-OS 98 +/- 1% vs. 99 +/- 1% vs. 94 +/- 2%, P = 0.056). Multivariate analysis demonstrated an impact of INSS stage on EFS only. IDRF were not shown to be significant for predicting EFS or OS. CONCLUSIONS: IDRF were useful in predicting risk and completeness of operation. IDRF failed as independent risk predictors in localized neuroblastoma. INSS more precisely identified patients with poor prognosis. PMID- 17914734 TI - Longitudinal assessment of health-related quality of life in preschool children with non-CNS cancer after the end of successful treatment. AB - BACKGROUND: The aim of the study was to access Health Related Quality of Life (HRQoL) in preschool cancer survivors during the first 3 years of continuous remission after the end of successful treatment, and to identify predictors of HRQoL. PROCEDURE: Parent-reported HRQoL was assessed in 53 preschool children treated successfully for cancer, using the TAPQOL and compared with norm data. Longitudinal mixed models analyses were performed to investigate to what extent demographic and medical variables and parental psychological distress were predictive of HRQoL over time. RESULTS: Two months after the end of successful cancer treatment, survivors showed significantly (P < 0.01) more problem behavior and anxiety, and scored significantly worse (P < 0.01) on sleeping, motor functioning, positive mood and liveliness than the norm. One year after the end of treatment survivors still showed significantly (P < 0.01) more anxiety and worse motor functioning. The level of HRQoL in survivors had normalized 2 and 3 years after the end of treatment. Longer duration of treatment, bad prognosis and greater parental psychological distress were associated with worse scores on the Physical Component Score of the TAPQOL. Medical variables and parental psychological distress were not associated with the Mental Component Score. CONCLUSIONS: Survivors adjusted well to the cancer experience and HRQoL improved with time. Despite overall resilience in survivors over time, physical as well as psychosocial monitoring in follow-up is recommended. Standard aftercare should preferably include psychosocial screening, education, and counseling directed at both survivors and parents. PMID- 17914736 TI - Chronic metastatic neuroblastoma. AB - The diversity of neuroblastoma and its clinical course depends on histology, biology and clinical features. We report a male presenting at 4 months of age with an abdominal mass and multiple subcutaneous nodules. The diagnosis was made by histological examination of a subcutaneous nodule and elevated urinary markers. The patient remained well during the subsequent 9 years. During that time no cytostatic treatment was given. Attempt to treat with cis-retinoic acid 10 years later did not result in any significant change of the clinical course. The patient has remained in good clinical condition for a 15-year observation period, having both progressing and regressing distant subcutaneous metastases. Skin nodules are the hallmarks of the indolent clinical course of the disease. We suggest the use of the "chronic neuroblastoma" as a term to describe patients with neuroblastoma showing indolent disease course over a very long period of time, but never achieving complete remission. PMID- 17914737 TI - Chemokine CXCL12 enhances proliferation in pre-B-ALL via STAT5 activation. AB - BACKGROUND: CXCL12 (SDF-1alpha) is a chemokine, which plays an important role in normal B-cell lymphopoesis, migration and homing to the bone marrow (BM) and previous studies have suggested a role for CXCL12 and its receptor CXCR4 in the pathogenesis of ALL. PURPOSE: CXCL12 levels in serum were evaluated from ALL children and controls. The biological effect of recombinant CXCL12 on primary leukaemic cells was investigated. Signalling via the CXCL12/CXCR4 axis was further characterized in an in vitro model using the pre-B leukaemic cell line Nalm-6. RESULTS: The serum level of CXCL12 in children at diagnosis of pre-B-ALL is significantly higher than in healthy children (4.8 (0-32) ng/ml vs. 0 (0-3.2) ng/ml, P < 0.001). After completed chemotherapy, CXCL12 decreases to levels comparable to those found in the control group. In addition, we found that recombinant CXCL12 enhances pre-B leukaemic cell proliferation in vitro. The CXCL12/CXCR4 axis is able to initiate functional signalling and we show that STAT5 is activated in CD19+ leukaemic cells from BM of ALL patients and in the leukaemic cell line Nalm-6. CONCLUSION: Our findings suggest that CXCL12 may have a role in leukaemic cell proliferation and survival during childhood ALL. PMID- 17914738 TI - Cutaneous zygomycosis in an infant with Pearson syndrome. PMID- 17914739 TI - Advanced small cell carcinoma of the ovary in a seventeen-year-old female, successfully treated with surgery and multi-agent chemotherapy. AB - Advanced small cell carcinoma of the ovary (FIGO stage III or IV) is a rare and usually lethal tumor seen in adolescents and young women. In pediatric patients with advanced disease, there have been only two case reports of successful therapy, we report a third patient, diagnosed at 17 years of age, with an abdominal mass and metastatic disease to regional and distant lymph nodes, who was successfully treated with surgery and intensive multi-agent chemotherapy. Imatinib, thalidomide, and celecoxib were also administered for up to 24 months following initial chemotherapy. She remains in remission 3 years from diagnosis. PMID- 17914740 TI - Pediatric orbital and ocular lymphomas. PMID- 17914741 TI - Isolated cutaneous anaplastic large cell lymphoma progressing to severe systemic disease with myocardial involvement and central nervous system infiltration. AB - Anaplastic large cell lymphoma (ALCL) is a rare tumor comprising around 10-15% of childhood lymphomas. We describe the case of a female who initially presented with localized skin disease associated with an insect bite. However, she subsequently relapsed with widespread systemic ALK-positive ALCL that included lymphoma deposits in the myocardium, a very rare manifestation. Her disease responded well to chemotherapy but she later developed a fatal relapse in the CNS. We also present data on an immune response to ALK, demonstrating a fluctuation in the levels of circulating antibodies to ALK corresponding to the different phases of her illness. PMID- 17914742 TI - Trigonal injection of botulinum toxin-A does not cause vesicoureteral reflux in neurogenic patients. AB - AIMS: We evaluated the effect of botulinum toxin type A (BTX-A) injections in the trigone on the antireflux mechanism and evaluated its short-term efficacy. MATERIALS AND METHODS: Between April and December 2006, 21 patients (10 men and 11 women) were prospectively evaluated. All were incontinent due to refractory NDO and underwent detrusor injection of 300 units of BTX-A, including 50 units into the trigone. Baseline and postoperative evaluation after eight weeks included cystogram, urinary tract ultrasound and urodynamics. RESULTS: At baseline, 20 patients had no vesicoureteral (VUR) and one had grade II unilateral VUR. Postoperative evaluation revealed no cases of de novo VUR and the patient with preinjection VUR had complete resolution of the reflux. Ultrasound showed 5 (23.8%) patients with hydronephrosis before BTX-A injection and only one (4.8%) at the followup evaluation (p=0.066). After treatment, 9 (42.8%) patients became dry, 11 (52.4%) were improved and one (4.8%) had no improvement. Improved patients received antimuscarinic treatment and 8 (38.1%) became dry, with a final total continence rate of 80.1%. Cystometric capacity increased from 271+/-92 to 390+/-189 ml (p=0.002), reflex volume varied from 241+/-96 to 323+/-201 ml (p=0.020) and maximum detrusor pressure reduced from 66+/-39 to 38+/-37 cm H(2)O (p<0.001). CONCLUSIONS: Our results confirm the safety of trigone injections of BTX-A in terms of development of VUR and upper urinary tract damage. Whether they are beneficial for patients with NDO or other causes of voiding dysfunction will need further studies. PMID- 17914743 TI - Cancer metastasis facilitated by developmental pathways: Sonic hedgehog, Notch, and bone morphogenic proteins. AB - This review will highlight the significance of three critical pathways in developmental biology and our emerging understanding of their roles in regulating tumor metastasis: Bone morphogenic protein (BMP), Notch and Sonic hedgehog (SHH). We will discuss parallels between their known roles in development and how these processes can be used by tumor cells to create microenvironments that enhance tumor metastasis. That tumor cells usurp pathways critical to the developing embryo is not surprising, as many of the normal developmental programs include processes that are also seen during tumor progression to a metastatic phenotype, including epithelial to mesenchymal transition (EMT), tissue specific morphogenesis, cellular motility and invasion. BMPs are involved in EMT, contribute to tissue specific morphogenesis, and are expressed in highly metastatic tumor cells. BMPs have also been hypothesized to have a role in the establishment of a pre-neoplastic niche. Notch and SHH facilitate neovascularization, angiogenesis, EMT and can contribute to the maintenance of highly-metastatic tumor stem cells. PMID- 17914744 TI - Intrauterine-diagnosed diastematomyelia. PMID- 17914745 TI - Identification and characterization of a novel yeast omega3-fatty acid desaturase acting on long-chain n-6 fatty acid substrates from Pichia pastoris. AB - A cDNA sequence putatively encoding a omega(3)-fatty acid desaturase gene was isolated from methylotrophic yeast Pichia pastoris GS115. The deduced amino acid sequence of this cloned cDNA showed high identity to known fungal omega(3)-fatty acid desaturases. Functional identification of this gene heterologously in Saccharomyces cerevisiae strain INVScl indicated that the deduced amino acid sequence exhibited omega(3)-fatty acid desaturase activity. The newly identified omega(3)-fatty acid desaturase, named Pp-FAD3, is novel because it showed broad n 6 fatty acid substrate specificity by its ability to convert all the 18-carbon and 20-carbon n-6 substrates examined to the corresponding n-3 fatty acids, with an approximately equivalent high conversion rate. Pp-FAD3 is the first known yeast omega(3)-fatty acid desaturase to act on long-chain n-6 fatty acid substrates. Heterologous expression of the newly identified omega(3) desaturase in different hosts will be an alternative method to increase the flow of n-6 fatty acid intermediates into their n-3 derivatives. PMID- 17914746 TI - Genome-wide analysis of coding DNA and amino acid variation in Saccharomyces cerevisiae. AB - The possible causes of variation on amino acid composition in the yeast Saccharomyces cerevisiae were investigated genome-wide. The results indicated that: (a) the base composition of coding DNA and amino acid composition was similar among all the chromosomes, which was in sharp contrast with the great varies of the composition of the individual's coding DNA and amino acid; (b) some amino acids (e.g. Cys and Trp) were not present in all the proteins; and (c) amino acid bias was associated with a base bias (in terms of A-, G-, C- and T rich codons). Based on the third rule and a proposed universal trend of amino acid gain and loss in protein evolution, the changing pattern of coding DNA was predicted to be T- and C-accruing, whereas A and G were consistently reducing. All these results held the potential to reveal precisely how DNA ongoing change has a major effect on the composition of proteins. PMID- 17914747 TI - The transcriptional inhibitor thiolutin blocks mRNA degradation in yeast. AB - Thiolutin is commonly used as a general inhibitor of transcription in yeast. It has been used to calculate mRNA decay rates by stopping the transcription and then determining the relative abundance of individual mRNAs at different times after inhibition. We report here that thiolutin is also an inhibitor of mRNA degradation, and thus its use can lead to miscalculations of mRNA half-lives. The inhibition of mRNA decay seems to affect the mRNA degradation pathway without impeding poly(A) shortening, given that the decay rate of total poly(A) amount is not reduced by thiolutin. Moreover, the thiolutin-dependent inhibition of mRNA degradation has variable effects on different functional groups of genes, suggesting that they use various degradation pathways for their mRNAs. PMID- 17914748 TI - Membrane topology and post-translational modification of the Saccharomyces cerevisiae essential protein Rot1. AB - ROT1 is an essential gene that has been related to cell wall biosynthesis, the actin cytoskeleton and protein folding. In order to help to understand its molecular function, we carried out a characterization of the Rot1 protein. It is primarily located at the endoplasmic reticulum-nuclear membrane facing the lumen. Rot1 migrates more slowly than expected, which might suggest post-translational modification. Our results indicate that Rot1 is a protein that is neither GPI anchored nor O-glycosylated. In contrast, it is N-glycosylated. By a directed mutagenesis of several Asn residues, we identified that the protein is simultaneously glycosylated at N103, N107 and N139. Although the mutation of these three N sites is not lethal, cellular growth is impaired. Sequence analysis predicts a transmembrane domain at the C-terminus. This fragment affects neither the targeting of the Rot1 protein to the ER nor its N-glycosylation, although it is important for the anchoring of the protein to the membrane and for its functionality. The existence of a signal sequence at the N-terminus has been suggested. However, deletion of this fragment impedes neither translocation to the ER nor N-glycosylation, but it is required for cell viability. Finally, we found that Rot1 is translocated to the ER by an SRP-independent post translational mechanism which depends on Sec62. PMID- 17914749 TI - Expression of glycerol 3-phosphate dehydrogenase gene (CvGPD1) in salt-tolerant yeast Candida versatilis is stimulated by high concentrations of NaCl. AB - We cloned the glycerol 3-phosphate dehydrogenase (GPDH) gene (CvGPD1) from salt tolerant yeast Candida versatilis. When CvGPD1 was expressed in glycerol synthesis-deficient Saccharomyces cerevisiae cells, the salt tolerance of the recombinant strain was enhanced, and NADP(+)-dependent GPDH (EC 1.1.1.94), Cvgpd1p synthesis and recovery of glycerol synthesis were confirmed. The transcription of CvGPD1 in C. versatilis cells was stimulated by high concentrations of NaCl. The relationship between expression of CvGPD1 and growth of C. versatilis cells in the mash of Japanese seasonings (miso- and shoyu moromi) is also discussed. PMID- 17914750 TI - Validation of the PathFinder neurosurgical robot using a phantom. AB - BACKGROUND: Minimally invasive surgery was born out of recent advances in neuro imaging and stereotactic technology. As a result, the scale of neurosurgical procedures will soon be so small that it will not be within the ability of the most gifted and skilled neurosurgeons of today. Hence, neurosurgical robotics is the natural evolution in this field. The aim of this study was to evaluate the performance of a new robotic system in a neurosurgical phantom, comparing it to standard frame-based and frameless technology of today. METHODS: In total, 19 different targets were approached by two standard stereotactic frames, the Stealth Station frameless system and the robot. The CRW and the ZD stereotactic frames were used. The frameless system was the Stealth Station image guidance system. The phantom used was a replica of the human skull fitted with 10 surface and nine deep targets. RESULTS: The robotic system outperformed both frame-based and frameless systems in all experiments in this study. The application accuracies were: robot, 0.5 mm; stereotactic frames, 0.98 mm; and frameless system, 1.96 mm. CONCLUSIONS: The robotic system was as accurate as the stereotactic frame, but without technical restrictions and cumbersome manual adjustments. Furthermore, the robotic system had near-absolute geometric accuracy, was reliable to perform the same procedure over and over without tiresomeness, variation or boredom, and would be impervious to biohazards and hostile environments. PMID- 17914751 TI - Two- and four-dimensional echocardiography with B-flow imaging and spatiotemporal image correlation in prenatal diagnosis of isolated total anomalous pulmonary venous connection. AB - OBJECTIVES: To explore whether the use of four dimensional (4D) ultrasound examination with B-flow imaging and spatiotemporal image correlation (STIC) can supply additional information with respect to two-dimensional (2D) gray-scale and color Doppler echocardiography in the prenatal characterization of isolated total anomalous pulmonary venous connection (TAPVC). METHODS: The study population comprised a group of three TAPVC fetuses that had been examined exclusively by conventional echocardiography, and a group of four additional cases initially identified by conventional echocardiography and examined further by 4D ultrasonography; a thorough postnatal work-up was available for all fetuses. RESULTS: At our center, isolated TAPVC was found in seven of 1040 fetuses with cardiac defects (0.67%). Anomalous drainage was supracardiac to the innominate vein in three cases, cardiac to the coronary sinus in two, and infracardiac to the portal vein in the remaining two cases. An evident asymmetry between left heart and right heart structures was observed in 4/7 cases. The confluence of the anomalous pulmonary veins (PVs) was visualized in 4/7 cases and the connecting vertical vein was identified in 3/5 cases at 2D echocardiography. 4D ultrasound imaging with B-flow imaging and STIC clearly visualized the anomalous PV confluence and the draining vertical vein in all four cases examined. CONCLUSION: 2D and color Doppler echocardiography appears to diagnose reliably TAPVC, albeit with some limitations in thorough assessment of the pathology, depending on the anatomy of the defect, on the technical adequacy of the equipment used and on the experience of the operator. 4D ultrasound examination with B-flow imaging and STIC is apparently able to facilitate identification of the anatomical features of TAPVC, thus supplying additional information over that provided by 2D fetal sonography. PMID- 17914753 TI - Centrifugal microfluidics with integrated sensing microdome optodes for multiion detection. AB - An array of four sensing microdome optodes (potassium, sodium, calcium, and chloride) was incorporated into a centrifugal microfluidics platform to obtain a multiion analysis system. The behavior of each sensing microdome was in good agreement with a theoretical model describing the response. The selectivity of each optode over common interfering ions was established and was used to identify calibrant solutions that can be employed for the simultaneous calibration of all four optodes without significant cross-interference. The microfluidic platform was designed to facilitate both three-point calibration of the optodes and triplicate analysis of a sample within a single run, which increases the accuracy of the determination. The optimized microfluidic system was used to determine simultaneously the concentration of potassium, sodium, calcium, and chloride in aquarium water (with the composition of Lake Tanganyika water) with less than 6% error. The simple process of fabrication of these microdomes and their incorporation into a centrifugal microfluidic platform should facilitate the development of portable ion-sensing analysis systems. PMID- 17914752 TI - Structural characterization of photodegradation products of enalapril and its metabolite enalaprilat obtained under simulated environmental conditions by hybrid quadrupole-linear ion trap-MS and quadrupole-time-of-flight-MS. AB - In the environment, organic micropollutants such as pharmaceuticals can be degraded via various biotic and abiotic transformation routes. In surface waters, for example, photodegradation may constitute a relevant natural attenuation process for drug residues that have been discharged from sewage treatment facilities. In the present work, the photochemical fate of the prodrug enalapril (376 Da, C20H28N2O5) and its active metabolite enalaprilat (348 Da, C18H24N2O5), a hypotensive cardioprotector previously reported to occur in contaminated rivers, was investigated in aqueous media under the influence of irradiation generated by a sunlight simulator. The experiments yielded three detectable photodegradates for enalapril (346 Da, 2 x 207 Da) whereas the photolysis of enalaprilat went hand in hand with the intermittent buildup of one photodegradate (304 Da). Fragmentation patterns of the parent compounds were established on a hybrid quadrupole-linear ion trap-mass spectrometer exploiting its MS3 capabilities. Accurate mass measurements recorded on a hybrid quadrupole-time-of flight instrument in MS/MS mode allowed us to propose elemental compositions for the molecular ions of the degradates (346 Da, C19H26N2O4; 207 Da, C12H17NO2; 304 Da, C17H24N2O3) as well as of their fragment ions. Based on these complementary data sets from the two distinct mass spectrometric instruments, plausible structures were postulated for the four photodegradates. The compounds formed by enalapril corresponded to the loss of formaldehyde out of the proline residue (346 Da), cleavage of the central amide bond (207 Da) followed by migration of the ethylester side chain (207 Da) while decarboxylation of the free carboxylic acid was described for enalaprilat (304 Da). The study emphasized the potential of sunlight for breaking down an environmentally relevant drug and its metabolite. PMID- 17914754 TI - Single-molecule detection of surface-hybridized human papilloma virus DNA for quantitative clinical screening. AB - We present an improved method to quantify viral DNA in human cells at the single- molecule level. Human papilloma virus (HPV)-16 DNA was hybridized to probes that were covalently bound to a glass surface and detected with a single-molecule imaging system. In the single-probe mode, the whole genome and target DNA were fluorescently labeled before hybridization. In the dual-probe mode, a second probe was introduced that has a fluorescently labeled 1-kb DNA strand connected to the 50-nt probe sequence. With the single-probe method, the detection limit was 0.7 copy/cell, which was similar to that reported in a flow system earlier. With the dual-probe method, the linear dynamic range covers 1.44-7000 copies/cell, which is typical of early infection to near-cancer stages. Both methods were applied to cell line samples with known HPV-16 infection, and the result showed a good match with the reported viral load. DNA from cervical cells, collected with the Pap smear sampling method, was spiked with HPV-16 DNA and submitted to this assay to show compatibility with conventional sampling methods. The dual-probe method was further tested with a crudely prepared sample. The cells were heat lyzed and spun down, and the supernatant was immediately submitted to hybridization. Even with reduced hybridization efficiency caused by the interference of cellular materials, we were still able to differentiate infected cells with 600 copies/cell from healthy cells. PMID- 17914755 TI - Microcantilever-based sensors: effect of morphology, adhesion, and cleanliness of the sensing surface on surface stress. AB - The surface stress response of micromechanical cantilever-based sensors was studied as a function of the morphology, adhesion, and cleanliness of the gold sensing surface. Two model systems were investigated: the adsorption of alkanethiol self-assembled monolayers at the gas-solid interface and the potential-controlled adsorption of anions at the liquid-solid interface. The potential-induced surface stress, on a smooth and continuous polycrystalline Au(111)-textured microcantilever in 0.1 M HClO4, is in excellent agreement with macroscopic Au(111) single-crystal electrode results. It is shown that ambient contaminants on the sensing surface dramatically alter the surface stress potential response. This observation can be misinterpreted as evidence that for polycrystalline Au(111) microcantilever electrodes, surface stress is dominated by surface energy change. Results for anions adsorption on gold are in contrast to the gas-phase model system. We demonstrate that the average grain size of the gold sensing surface strongly influences the magnitude of the surface stress change induced by the adsorption of octanethiol. A 25-fold amplification of the change in surface stress is observed on increasing the average gold grain size of the sensing surface from 90 to 500 nm. PMID- 17914756 TI - Structural induced control of energy transfer within Zn(II)-porphyrin dendrimers. AB - We report on a study of singlet-singlet annihilation kinetics in a series of Zn(II)-porphyrin-appended dendrimers, where the energy transfer efficiency is significantly improved by extending the molecular chain that connects the light harvesting chromophores to the dendrimeric backbone with one additional carbon. For the largest dendrimer having 64 Zn(II)-porphyrins, only approximately 10% of the excitation intensity is needed in order to observe the same extent of annihilation in the dendrimers with the additional carbon in the connecting chain as compared to those without. Complete annihilation, until only one chromophore remains excited, now occurs within subunits of seven chromophores, when half of the chromophores are excited. The improvement of the annihilation efficiency in the largest dendrimer with 64 porphyrins can be explained by the presence of a the two-step delayed annihilation process, involving energy hopping from excited to nonexcited chromophores prior to annihilation. In the smallest dendrimer with only four chromophores, delayed annihilation is not present, since the direct annihilation process is more efficient than the two-step delayed annihilation process. As the dendrimer size increases and the chances of originally exciting two neighboring chromophores decreases, the delayed annihilation process becomes more visible. The additional carbon, added to the connecting chain, results in more favorable chromophore distances and orientations for energy hopping. Hence, the improved energy transfer properties makes the Zn(II)-porphyrin-appended dendrimers with the additional carbon promising candidates as light-harvesting antennas for artificial photosynthesis. PMID- 17914757 TI - Theoretical study on the assembly and stabilization of a magic cluster Al4N-. AB - We report the first attempt to assemble the magic cluster Al4N- on the basis of the density functional theory calculations on a series of pi-stacked dimers (Al4N )2, sandwich-like compounds [D(Al4N)M]q- (where D = Al4N-, Cp-(C5H5-); M = Li, Na, K, Be, Mg, Ca) and extended compounds (Cp-)m(Li+)n(Al4N-)o (where m, n, and o are integers). For the six metals, the magic Al4N- can only be assembled and grow up in our newly proposed "hetero-decked sandwich" scheme (e.g., [CpM(Al4N)]q-) so as to avoid cluster fusion. The ground-state hetero-decked sandwich species (Cp )(M)q+(Al4N)- (M = Li, Na, K, q = 1; M = Be, Mg, Ca, q = 2) and the extended sandwich species (Cp-)m(Li+)n(Al4N-)o are mainly ionically bonded, cluster assembled "polyatomic molecules", grown from the combination of Cp-, M-atoms, and Al4N-. As a prototype for ionic bonding involving intact Al4N- subunits, [CpM(Al4N)]q- may be a stepping stone toward forming ionic, cluster-assembled AlN compounds. PMID- 17914758 TI - Energetics and dynamics of the fragmentation reactions of protonated peptides containing methionine sulfoxide or aspartic acid via energy- and time-resolved surface induced dissociation. AB - The surface-induced dissociation (SID) of six model peptides containing either methionine sulfoxide or aspartic acid (GAILM(O)GAILR, GAILM(O)GAILK, GAILM(O)GAILA, GAILDGAILR, GAILDGAILK, and GAILDGAILA) have been studied using a specially configured Fourier transform ion-cyclotron resonance mass spectrometer (FT-ICR MS). In particular, we have investigated the energetics and dynamics associated with (i) preferential cleavage of the methionine sulfoxide side chain via the loss of CH3SOH (64 Da), and (ii) preferential cleavage of the amide bond C-terminal to aspartic acid. The role of proton mobility in these selective bond cleavage reactions was examined by changing the C-terminal residue of the peptide from arginine (nonmobile proton conditions) to lysine (partially mobile proton conditions) to alanine (mobile proton conditions). Time- and energy-resolved fragmentation efficiency curves (TFECs) reveal that selective cleavages due to the methionine sulfoxide and aspartic acid residues are characterized by slow fragmentation kinetics. RRKM modeling of the experimental data suggests that the slow kinetics is associated with large negative entropy effects and these may be due to the presence of rearrangements prior to fragmentation. It was found that the Arrhenius pre-exponential factor (A) for peptide fragmentations occurring via selective bond cleavages are 1-2 orders of magnitude lower than nonselective peptide fragmentation reactions, while the dissociation threshold (E0) is relatively invariant. This means that selective bond cleavage is kinetically disfavored compared to nonselective amide bond cleavage. It was also found that the energetics and dynamics for the preferential loss of CH3SOH from peptide ions containing methionine sulfoxide are very similar to selective C-terminal amide bond cleavage at the aspartic acid residue. These results suggest that while preferential cleavage can compete with amide bond cleavage energetically, dynamically, these processes are much slower compared to amide bond cleavage, explaining why these selective bond cleavages are not observed if fragmentation is performed under mobile proton conditions. This study further affirms that fragmentation of peptide ions in the gas phase are predominantly governed by entropic effects. PMID- 17914759 TI - Substituent effects on the energetics and aromaticity of aminomethylbenzoic acids. AB - The standard (p0 = 0.1 MPa) molar enthalpies of combustion of six aminomethylbenzoic acids were measured at T = 298.15 K by static bomb calorimetry. With these values, the standard molar enthalpies of formation in the crystalline state were obtained. Combining these results with the standard molar enthalpies of sublimation, the standard molar enthalpies of formation in the gaseous phase were derived. For the 10 possible isomers, the obtained experimental results were compared to and correlated with the relative stability obtained by ab initio calculations at the B3LYP/6-311++G(d,p) level of theory. Seeking a better understanding of the aromatic behavior and energetics of aminomethylbenzoic acids in the gas phase, calculations of NICS values, HOMA indices, and dihedral angles between the aromatic carbon and the amino group, Phi(Ar-NHH), were also performed computationally. The significant differences observed in the energetics, as well as in the NICS values, HOMA indices, and Phi(Ar-NHH) dihedral angles for these 10 isomers suggest a strong dependency on the identity and relative position of the three substituents on the benzene ring. This study points out a marked tendency for a decrease of the ring aromaticity, accompanied by an increase in the respective system stability, as the conjugation between the substituents becomes more extensive. PMID- 17914760 TI - Accurate predictions of crystal densities using quantum mechanical molecular volumes. AB - A quantum mechanically based procedure for estimation of crystal densities of neutral and ionic crystals is presented. In this method, volumes within 0.001 electrons/bohr3 isosurfaces of electron density for the constituent isolated neutral and ionic molecules are calculated to define the molecular volume or formula unit volumes used in predicting the crystal density. The B3LYP density functional theory in conjunction with the 6-31G** basis set were employed to generate the electron densities. The suitability of this method of crystal density prediction was assessed by subjecting a large number (289) of molecular and ionic crystals to the procedure and comparing results with experimental information. The results indicate that, for neutral molecular crystals, the root mean-square (rms) deviation from experiment is within 4%, whereas the rms deviation is somewhat larger for the 71 ionic crystals evaluated (within 5%). PMID- 17914761 TI - Longitudinal nuclear spin relaxation of ortho- and para-hydrogen dissolved in organic solvents. AB - The longitudinal relaxation time of ortho-hydrogen (the spin isomer directly observable by NMR) has been measured in various organic solvents as a function of temperature. Experimental data are perfectly interpreted by postulating two mechanisms, namely intramolecular dipolar interaction and spin-rotation, with activation energies specific to these two mechanisms and to the solvent in which hydrogen is dissolved. This permits a clear separation of the two contributions at any temperature. Contrary to the self-diffusion coefficients at a given temperature, the rotational correlation times extracted from the dipolar relaxation contribution do not exhibit any definite trend with respect to solvent viscosity. Likewise, the spin-rotation correlation time obeys Hubbard's relation only in the case of hydrogen dissolved in acetone-d6, yielding in that case a spin-rotation constant in agreement with literature data. Concerning para hydrogen, which is NMR-silent, the only feasible approach is to dissolve para enriched hydrogen in these solvents and to follow the back-conversion of the para isomer into the ortho-isomer. Experimentally, this conversion has been observed to be exponential, with a time constant assumed to be the relaxation time of the singlet state (the spin state of the para-isomer). A theory, based on intermolecular dipolar interactions, has been worked out for explaining the very large values of these relaxation times which appear to be solvent-dependent. PMID- 17914762 TI - Efflorescence relative humidity of mixed sodium chloride and sodium sulfate particles. AB - We study the efflorescence relative humidity (ERH) of particles composed of sodium chloride and sodium sulfate. Both experimental and theoretical investigations are conducted to explore the effects of particle size and mixing ratios between two inorganic materials on ERH. A previously developed theoretical model (Gao et al. J. Phys. Chem. A 2006, 110, 7602; ref 1) is applied as the framework to build a formulation assuming that one salt nucleates much faster than the other, and the critical nuclei formation of the former controls the rate of efflorescence. The predicted ERHs agree favorably with the experimental data, except for particles containing Na2SO4 in a mole fraction of around 0.25. At this composition, our model underestimates the ERH, indicating certain factors involved in the efflorescent processes that are overlooked in our formulation. Relative to particles larger than 40 nm, the Kelvin effect more significantly affects particles smaller than this size. PMID- 17914764 TI - Computational s-block thermochemistry with the correlation consistent composite approach. AB - The correlation consistent composite approach (ccCA) is a model chemistry that has been shown to accurately compute gas-phase enthalpies of formation for alkali and alkaline earth metal oxides and hydroxides (Ho, D. S.; DeYonker, N. J.; Wilson, A. K.; Cundari, T. R. J. Phys. Chem. A 2006, 110, 9767). The ccCA results contrast to more widely used model chemistries where calculated enthalpies of formation for such species can be in error by up to 90 kcal mol-1. In this study, we have applied ccCA to a more general set of 42 s-block molecules and compared the ccCA DeltaHf values to values obtained using the G3 and G3B model chemistries. Included in this training set are water complexes such as Na(H2O)n+ where n = 1 - 4, dimers and trimers of ionic compounds such as (LiCl)2 and (LiCl)3, and the largest ccCA computation to date: Be(acac)2, BeC10H14O4. Problems with the G3 model chemistries seem to be isolated to metal-oxygen bonded systems and Be-containing systems, as G3 and G3B still perform quite well with a 2.7 and 2.6 kcal mol-1 mean absolute deviation (MAD), respectively, for gas-phase enthalpies of formation. The MAD of the ccCA is only 2.2 kcal mol-1 for enthalpies of formation (DeltaHf) for all compounds studied herein. While this MAD is roughly double that found for a ccCA study of >350 main group (i.e., p block) compounds, it is commensurate with typical experimental uncertainties for s-block complexes. Some molecules where G3/G3B and ccCA computed DeltaHf values deviate significantly from experiment, such as (LiCl)3, NaCN, and MgF, are inviting candidates for new experimental and high-level theoretical studies. PMID- 17914763 TI - Structural and optical isomers of nonamethoxy cyclotriveratrylene: separation and physical characterization. AB - The paper concerns the structural and optical isomers of nonamethoxy tribenzocyclononene (compound 1). In the first part of the paper it is shown that 1 exists in two structural isomers: a rigid crown (c-1) with C3 symmetry and a flexible saddle (s-1) with C1 symmetry. The latter, not previously known, can be prepared from the as-synthesized c-1 by quenching a hot solution (or the melt) followed by HPLC separation. The crown/saddle equilibrium, isomerization kinetics, and associated thermodynamic parameters in various organic solvents are reported. Carbon-13 MAS NMR, X-ray diffraction, and differential scanning calorimetry (DSC) of polycrystalline c-1 and s-1 racemates are also reported. The different melting points of the isomers and their rapid isomerization in the melt result in unconventional DSC thermograms involving multiple endothermic and exothermic transitions. The second part of the paper concerns the chiral properties of 1. Both the saddle and crown isomers are structurally chiral, but due to the fast pseudorotation of s-1 in solution, it cannot be separated into its enantiomers. Those of c-1 were separated by HPLC using a chiral column. Their X-ray structure and melting points differ considerably from those of the racemate. This and their fast racemization in the melt lead to complex DSC thermograms with multiple transitions. Solutions of the neat enantiomers exhibit a relatively small specific optical rotation. In the UV they show circular dichroism for the B1u and B2u transitions, with the latter exhibiting a clear couplet structure. Infrared and vibrational circular dichroism spectra of the enantiomers in solution are reported. Comparison of these spectra with quantum mechanical simulations provides unambiguous identification of the enantiomers. PMID- 17914765 TI - Pump-degenerate four wave mixing as a technique for analyzing structural and electronic evolution: multidimensional time-resolved dynamics near a conical intersection. AB - Pump-degenerate four wave mixing (pump-DFWM) is used to simultaneously study the early events in structural and electronic population dynamics of the non adiabatic passage between two excited electronic states. After the precursor state S2 is populated by an initial pump beam, a DFWM sequence is set resonant with the S1 --> Sn transition on the successor state S1. The information obtained by pump-DFWM is two-fold: by scanning the delay between the initial pump and the DFWM sequence, the evolution of the individual excited-state modes is observed with a temporal resolution of 20 fs and a spectral resolution of 10 cm-1. Additionally, pump-DFWM yields information on electronic population dynamics, resulting in a comprehensive description of the S2 --> S1 internal conversion. As a system in which the interplay between structural and electronic evolution is of great interest, all-trans-beta-carotene in solution was chosen. The pump-DFWM signal is analyzed for different detection wavelengths, yielding results on the ultrafast dynamics between 1Bu+ (S2) and 2Ag- (S1). The process of vibrational cooling on S1 is discussed in detail. Furthermore, a low-lying vibrationally hot state is excited and characterized in its spectroscopic properties. The combination of highly resolved vibrational dynamics and simultaneously detected ultrafast electronic state spectroscopy gives a complete picture of the dynamics near a conical intersection. Because pump-DFWM is a pure time domain technique, it offers the prospect of coherent control of excited-state dynamics on an ultrafast time scale. PMID- 17914766 TI - Reactions of radicals with hydrolyzed Bi(III) ions: a pulse radiolysis study. AB - Radiolytic reduction of BiOClO4 in aqueous solutions leads to the formation of bismuth clusters and larger nanoparticles. The mechanisms of redox reactions of the polycationic Bi(III) species that exist in the solution were investigated with pulse radiolysis. The kinetic and spectral properties of the transients formed by the reaction of these species with the primary radicals from water radiolysis are reported. The single-electron reduction product, Bi9(OH)224+, absorbs at lambdamax = 273 nm, while the OH adduct, Bi9(OH)235+, has a broad absorption spectrum with a maximum at 280 nm and a shoulder at 420 nm. Several rate constants were measured: k (e-aq + Bi9(OH)225+) = 1.2 x 1010 M-1 s-1 and k (OH + Bi9(OH)225+) = 1.5 x 109 M-1 s-1. The reduced species, Bi9(OH)224+ further reacts with (CH3)2COH radicals, but not with CH2C(CH3)2OH radicals from t butanol, to produce a doubly reduced polynuclear species. A few reactions of the reduction of the Bi salt in the presence of poly(acrylic acid) are also described. In the presence of the polymer, a metal-polymer complex is formed prior to the irradiation, and the reduction reactions are significantly slowed down. PMID- 17914767 TI - Density functional theory study on vibrational circular dichroism as a tool for analysis of intermolecular systems: (1:1) cysteine-water complex conformations. AB - This paper presents a discussion of the interaction energies for selected conformers of chiral l-cysteine and their (1:1) complexes with water at the B3LYP/aug-cc-pVDZ level. From among more than forty calculated 1:1 complexes three groups of complexes were singled out and examined by the B3LYP/aug-cc-pVDZ calculated vibrational circular dichroism (VCD) spectra. On the basis of analysis of the nu(OmicronEta) and nu(NuEta) and beta(OH2) and beta(NH2) ranges, the VCD spectra were found to be sensitive to conformational changes and water arrangement in cysteine complexes, and to be especially useful for discriminating between different chiral forms of intermolecular hydrogen-bonding complexes. In particular, we show that the VCD modes of an achiral water molecule after complex formation acquire significant rotational strengths whose signs change in line with the geometry of the complex. Moreover, for some water arrangements the VCD spectra can be sensitive to water-wagging conformers and, in temperatures low enough, the intensive nu(OmicronEtaWfree) and beta(H2O) VCD bands may be sufficiently separated to be splitted into pair of oppositely directed bands. PMID- 17914768 TI - Significance of ammonia in growth of atmospheric nanoclusters. AB - We apply accurate quantum chemistry methods to study the thermochemistry of molecular clusters containing ammonia, water, and sulfuric acid and investigate initial reaction steps in atmospheric nucleation by calculating free energies for the related reactions. The results indicate that ammonia is a key reactant enhancing the growth of small water-sulfuric acid clusters in atmospheric conditions. The role of ammonia becomes significant when the nanoclusters contain more than one or two sulfuric acid molecules. This implies a lower limit of 1:3 for the NH3/H2SO4 mole ratio of atmospheric sulfuric acid-water-ammonia clusters. PMID- 17914769 TI - Extension of the self-consistent-charge density-functional tight-binding method: third-order expansion of the density functional theory total energy and introduction of a modified effective coulomb interaction. AB - The standard self-consistent-charge density-functional-tight-binding (SCC-DFTB) method (Phys. Rev. B 1998, 58, 7260) is derived by a second-order expansion of the density functional theory total energy expression, followed by an approximation of the charge density fluctuations by charge monopoles and an effective damped Coulomb interaction between the atomic net charges. The central assumptions behind this effective charge-charge interaction are the inverse relation of atomic size and chemical hardness and the use of a fixed chemical hardness parameter independent of the atomic charge state. While these approximations seem to be unproblematic for many covalently bound systems, they are quantitatively insufficient for hydrogen-bonding interactions and (anionic) molecules with localized net charges. Here, we present an extension of the SCC DFTB method to incorporate third-order terms in the charge density fluctuations, leading to chemical hardness parameters that are dependent on the atomic charge state and a modification of the Coulomb scaling to improve the electrostatic treatment within the second-order terms. These modifications lead to a significant improvement in the description of hydrogen-bonding interactions and proton affinities of biologically relevant molecules. PMID- 17914770 TI - Direct characterization of hydrogen peroxide bleached thermomechanical pulp using spectroscopic methods. AB - The effects of thermomechanical pulp (TMP) bleaching with hydrogen peroxide under acidic and alkaline conditions were studied using different spectroscopic analytical methods. The results of hydroxyl radical determination in bleaching solutions, analyses of carbonyl and carboxyl groups contents in the pulp, and the cellulose fiber surface analysis by X-ray photoelectron spectroscopy (XPS) elucidate the chemistry of the hydrogen peroxide treatment. Diffuse reflectance laser flash photolysis (DRLFP) method showed the differences in the photochemical behavior that reflect the changes of the chromophoric system after the preliminary peroxide bleaching stage under acidic conditions. Fourier transform infrared (FTIR) spectroscopy confirmed the non-delignifying character of the bleaching process. Suppression of carbonyl and formation of carboxyl groups in the case of the two-stage peroxide bleaching performed in the presence of catalysts and stabilizers was also confirmed. FT-Raman studies showed the removal of coniferaldehyde groups after treatment under acidic and alkaline conditions. PMID- 17914771 TI - Thermochemical studies of benzoylnitrene radical anion: the N-H bond dissociation energy in benzamide in the gas phase. AB - The thermochemical properties of benzoylnitrene radical anion, C6H5CON-, were determined by using a combination of energy-resolved collision-induced dissociation (CID) and proton affinity bracketing. Benzoylnitrene radical anion dissociates upon CID to give NCO- and phenyl radical with a dissociation enthalpy of 0.85 +/- 0.09 eV, which is used to derive an enthalpy of formation of 33 +/- 9 kJ/mol for the nitrene radical anion. Bracketing studies with the anion indicate a proton affinity of 1453 +/- 10 kJ/mol, indicating that the acidity of benzamidyl radical, C6H5CONH, is between those of benzamide and benzoic acid. Combining the measurements gives an enthalpy of formation for benzamidyl radical of 110 +/- 14 kJ/mol and a homolytic N-H bond dissociation energy in benzamide of 429 +/- 14 kJ/mol. Additional thermochemical properties obtained include the electron affinity of benzamidyl radical, the hydrogen atom affinity of benzoylnitrene radical anion, and the oxygen anion affinity of benzonitrile. PMID- 17914772 TI - Gold nanoparticles decorated with oligo(ethylene glycol) thiols: protein resistance and colloidal stability. AB - The interactions between proteins and gold colloids functionalized with protein resistant oligo(ethylene glycol) (OEG) thiol, HS(CH2)11(OCH2CH2)6OMe (EG6OMe), in aqueous solution have been studied by small-angle X-ray scattering (SAXS) and UV vis spectroscopy. The mean size, 2R, and the size distribution of the decorated gold colloids have been characterized by SAXS. The monolayer-protected gold colloids have no correlations due to the low volume fraction in solution and are stable in a wide range of temperatures (5-70 degrees C), pH (1.3-12.4), and ionic strength (0-1.0 M). In contrast, protein (bovine serum albumin) solutions with concentrations in the range of 60-200 mg/mL (4.6-14.5 vol %) show a pronounced correlation peak in SAXS, which results from the repulsive electrostatic interaction between charged proteins. These protein interactions show significant dependence on ionic strength, as would be expected for an electrostatic interaction (Zhang et al. J. Phys. Chem. B 2007, 111, 251). For a mixture of proteins and gold colloids, the protein-protein interaction changes little upon mixing with OEG-decorated gold colloids. In contrast, the colloid-colloid interaction is found to be strongly dependent on the protein concentration and the size of the colloid itself. Adding protein to a colloidal solution results in an attractive depletion interaction between functionalized gold colloids, and above a critical protein concentration, c*, the colloids form aggregates and flocculate. Adding salt to such mixtures enhances the depletion effect and decreases the critical protein concentration. The aggregation is a reversible process (i.e., diluting the solution leads to dissolution of aggregates). The results also indicate that the charge of the OEG self-assembled monolayer at a curved interface has a rather limited effect on the colloidal stabilization and the repulsive interaction with proteins. PMID- 17914774 TI - Simulations of the absorption and fluorescence of indole in aqueous solution and at a nonpolar/polar interface. AB - Theoretical results are presented on the absorption and fluorescence of indole in aqueous solution as well as at the air/water surface. We use a combined quantum chemical statistical mechanical model with explicit solvent. An approximate ab initio complete active space self-consistent field description of the indole molecule is used, coupled to a discrete polarizable water medium. From the bulk simulations, strong support is found for the interchange mechanism, which explains the unusual solvent shift of the fluorescence of indole or tryptophan in a polar surrounding by a solvent induced switch of the fluorescing state. Two mechanisms are given to explain the different shifts for indole at the interface. First, a dielectric depletion effect, which is expected from the reduction of the amount of polar media. Second, an interface-specific effect, which derives from the stronger hydrogen bond formation at the surface. The latter effect acts to increase the shift for both absorption and emission at the surface as compared to the bulk. From these results, the intrinsic probe photophysics of tryptophan in proteins is discussed in terms of the properties of the protein/solvent interface and the orientation of the amino acid. PMID- 17914773 TI - Gas-phase spectroscopy of protonated 3-OH kynurenine and argpyrimidine. comparison of experimental results to theoretical modeling. AB - The aging process of the human lens is associated with accumulation of chromophores and fluorophores that impair visual function. In the present study, we examined the photodissociation of 3-OH-kynurenine and argpyrimidine. Furthermore, absorption spectra obtained in gas phase using an electrostatic ion storage ring were studied as gas phase absorption have been shown to be more similar to the in vivo condition than absorption spectra obtained in the liquid phase. Experimental results were compared to theoretical modeling using the multistate, multireference perturbation theory approach combined with advanced molecular modeling tools to account for the solvent effects and to provide direct support for band assignments. Absorption maxima were determined both experimentally and theoretically and significant differences between the two chromophores were found. In particular, 3-OH-kynurenine demonstrated a blue-shift of more than 130 nm in the aqueous phase compared to the gas-phase due to the existence of different 3-OH-kynurenine conformers, which are stable under different conditions and originate from the interplay between intra- and intermolecular interactions. Photodissociation of argpyrimidine and 3-OH kynurenine was observed in vacuum thus confirming the results previously obtained in liquid phase demonstrating that the photodestruction takes place in both media. PMID- 17914775 TI - Signature OH absorption spectrum from cluster models of solvation: a solvent-to solute charge transfer state. AB - Ab initio electronic structure theory calculations on cluster models support the characterization of the signature absorption spectrum of a solvated hydroxyl OH radical as a solvent-to-solute charge transfer state modulated by the hydrogen bonding environment. Vertical excited states in OH(H2O)n clusters (n = 0-7, 16) calculated at the TDDFT level of theory (with companion calculations at the EOM CCSD level of theory for n secondary > tertiary, in agreement with the trends gleaned from kinetic measurements. Analysis of the transition-state geometry reveals a shift toward more reagents-like structures in the primary --> secondary --> tertiary sequence, which concurs with the expectation from Hammond's postulate. Using the ab initio data, we calculate thermal rate constants via transition-state theory. Our highest-level calculations indicate that the room temperature relative reactivities of primary, secondary, and tertiary alkane sites in hydrogen-abstraction reactions by ground-state oxygen atoms are 1, 29, and 422, respectively. These results are used to interpret recent experiments on the reactions of O(3P) with liquid alkanes. PMID- 17914782 TI - Kinetics of the O + HCNO reaction. AB - The kinetics of the O + HCNO reaction were investigated by a relative rate technique using infrared diode laser absorption spectroscopy. Laser photolysis (355 nm) of NO2 was used to produce O atoms, followed by O atom reactions with CS2, NO2, and HCNO, and infrared detection of OCS product from the O + CS2 reaction. Analysis of the experiment data yields a rate constant of k1= (9.84 +/- 3.52) x 10-12 exp[(-195 +/- 120)/T)] (cm3 molecule-1 s-1) over the temperature range 298-375 K, with a value of k1 = (5.32 +/- 0.40) x 10-12 cm3 molecule-1 s-1 at 298 K. Infrared detection of product species indicates that CO producing channels, probably CO + NO + H, dominate the reaction. PMID- 17914783 TI - Ab initio molecular dynamical investigation of the finite temperature behavior of the tetrahedral Au19 and Au20 clusters. AB - Density functional molecular dynamics simulations have been carried out to understand the finite temperature behavior of Au19 and Au20 clusters. Au20 has been reported to be a unique molecule having tetrahedral geometry, a large HOMO LUMO energy gap, and an atomic packing similar to that of the bulk gold (Li, J.; et al. Science 2003, 299, 864). Our results show that the geometry of Au19 is exactly identical with that of Au20 with one missing corner atom (called a vacancy). Surprisingly, our calculated heat capacities for this nearly identical pair of gold clusters exhibit dramatic differences. Au20 undergoes a clear and distinct solid-like to liquid-like transition with a sharp peak in the heat capacity curve around 770 K. On the other hand, Au19 has a broad and flat heat capacity curve with continuous melting transition. This continuous melting transition turns out to be a consequence of a process involving a series of atomic rearrangements along the surface to fill in the missing corner atom. This results in a restricted diffusive motion of atoms along the surface of Au19 between 650 to 900 K during which the shape of the ground state geometry is retained. In contrast, the tetrahedral structure of Au20 is destroyed around 800 K, and the cluster is clearly in a liquid-like state above 1000 K. Thus, this work clearly demonstrates that (i) the gold clusters exhibit size sensitive variations in the heat capacity curves and (ii) the broad and continuous melting transition in a cluster, a feature that has so far been attributed to the disorder or absence of symmetry in the system, can also be a consequence of a defect (absence of a cap atom) in the structure. PMID- 17914784 TI - Electron transfer from axial ligand to S1- and S2-excited phosphorus tetraphenylporphyrin. AB - Photoinduced processes of a series of phosphorus tetraphenylporphyrin (PTPP) derivatives ([PTPP-(NHC6H4X)2]+Cl-, X = OCH3, CH3, H, Cl, CF3, and CN) have been investigated by using femtosecond laser flash photolysis mainly. PTPP with OH as an axial ligand showed S2 fluorescence upon excitation of the Soret band. The S2 fluorescence lifetime was estimated to be 1.5 ps. On the other hand, both S2 and S1 fluorescence bands of PTPP-(NHC6H4X)2 were difficult to observe, indicating the existence of an additional deactivation process such as charge separation (CS). From MO calculation and cyclic voltammetry, PTPP and the axial ligand are expected to act as an acceptor and a donor, respectively, upon excitation of PTPP. CS via the S2 state was confirmed during the femtosecond laser flash photolysis by observing the transient absorption of radical anion of PTPP. Furthermore, CS via the S1 state of PTPP was also observed. The CS rate via the S1 state was faster than that from the S2 state. The free energy dependence of the electron-transfer rates was discussed on the basis of Marcus theory. PMID- 17914785 TI - Discovery of 1-(4-methoxyphenyl)-7-oxo-6-(4-(2-oxopiperidin-1-yl)phenyl)-4,5,6,7 tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide (apixaban, BMS-562247), a highly potent, selective, efficacious, and orally bioavailable inhibitor of blood coagulation factor Xa. AB - Efforts to identify a suitable follow-on compound to razaxaban (compound 4) focused on modification of the carboxamido linker to eliminate potential in vivo hydrolysis to a primary aniline. Cyclization of the carboxamido linker to the novel bicyclic tetrahydropyrazolopyridinone scaffold retained the potent fXa binding activity. Exceptional potency of the series prompted an investigation of the neutral P1 moieties that resulted in the identification of the p methoxyphenyl P1, which retained factor Xa binding affinity and good oral bioavailability. Further optimization of the C-3 pyrazole position and replacement of the terminal P4 ring with a neutral heterocycle culminated in the discovery of 1-(4-methoxyphenyl)-7-oxo-6-(4-(2-oxopiperidin-1-yl)phenyl)-4,5,6,7 tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide (apixaban, compound 40). Compound 40 exhibits a high degree of fXa potency, selectivity, and efficacy and has an improved pharmacokinetic profile relative to 4. PMID- 17914786 TI - First example of phosphoramidate approach applied to a 4'-substituted purine nucleoside (4'-azidoadenosine): conversion of an inactive nucleoside to a submicromolar compound versus hepatitis C virus. AB - We report on the synthesis of the anti hepatitis C virus (HCV) agent 4' azidoadenosine (1) and the application of the phosphoramidate ProTide technology to this nucleoside. The synthesis of 1 was achieved through an epoxide intermediate followed by regio- and stereoselective ring opening by azidotrimethylsilane in the presence of a Lewis acid. Compound 1 did not inhibit HCV replication in cell culture at concentrations up to 0.1 mM. However, a submicromolar active agent could be derived from 1 by the application of the ProTide technology. All the phosphoramidates prepared were L-alanine derivatives with variations in the aryl moiety and in the ester part of the amino acid. The benzyl ester and the l-naphthyl phosphate (18) had the best activity in replicon assay. Phosphoramidates (18-21) achieved a significant improvement in antiviral potency over the parent nucleoside (1) with no increase in cytotoxicity. PMID- 17914788 TI - High-accuracy peptide mass fingerprinting using peak intensity data with machine learning. AB - For MALDI-TOF mass spectrometry, we show that the intensity of a peptide-ion peak is directly correlated with its sequence, with the residues M, H, P, R, and L having the most substantial effect on ionization. We developed a machine learning approach that exploits this relationship to significantly improve peptide mass fingerprint (PMF) accuracy based on training data sets from both true-positive and false-positive PMF searches. The model's cross-validated accuracy in distinguishing real versus false-positive database search results is 91%, rivaling the accuracy of MS/MS-based protein identification. PMID- 17914789 TI - Role of locked nucleic acid modified complementary strand in quadruplex/Watson Crick duplex equilibrium. AB - In the human genome, the G-rich sequences that form quadruplexes are present along with their C-rich complementary strands; this suggests the existence of equilibrium between a quadruplex and a Watson-Crick duplex which allows the execution of their respective biological functions. We have investigated the sensitivity of this equilibrium to pharmacological agents by employing locked nucleic acid (LNA) modified complementary strands, and demonstrated successful invasion of the stable telomeric quadruplex d[(G(3)TTA)(3)G(3)]. Fluorescence, UV, ITC, and SPR studies were performed to understand the binding process involving the preformed quadruplex and LNA-modified complementary strands compared with that involving the unmodified complementary strand. Our data indicate that LNA modifications in the complementary strand shift the equilibrium toward the duplex state. These modifications confer increased thermodynamic stability to the duplex and increase the magnitude of relative free energy (DeltaDeltaG degrees) difference between duplex and quadruplex, thus favoring the predominance of duplex population over quadruplex. This superior ability of LNA modified complementary strand can be exploited to pave an exploratory approach in which it hybridizes to a telomeric quadruplex and drives duplex formation, and inhibits the recognition of 3' G-rich overhang by RNA template of telomerase which guides telomere extension. PMID- 17914790 TI - Effects of polar group saturation on physical gelation of amphiphilic polymer solutions. AB - Monte Carlo simulation on the basis of the comblike coarse grained nonpolar/polar (NP) model has been carried out to study the polar group saturation effect on physical gelation of amphiphilic polymer solutions. The effects of polar group saturation due to hydrogen bonding or ion bridging on the sol-gel phase diagram, microstructure of aggregates, and chain conformation of amphiphilic polymer solutions under four different solvent conditions to either the nonpolar backbone or the polar side chain in amphiphilic polymer chains have been investigated. It is found that an increase of polar group saturation results in a monotonically decreased critical concentration of gelation point, which can be qualitatively supported by the dynamic rheological measurements on pectin aqueous solutions. Furthermore, various solvent conditions to either the backbone or the side chain have significant impact on both chain conformation and microstructure of aggregates. When the solvent is repulsive to the nonpolar backbone but attractive to the polar side chain, the polymer chains are collapsed, and the gelation follows the mechanism of colloidal packing; at the other solvent conditions, the gelation follows the mechanism of random aggregation. PMID- 17914791 TI - Spectrophotometric and calorimetric titration studies on molecular recognition of camphor and borneol by nucleobase-modified beta-cyclodextrins. AB - A series of modified beta-cyclodextrins with nucleobase substituents, that is, mono(6-ade-6-deoxy)-beta-cyclodextrin (2) and mono(6-ura-6-deoxy)-beta cyclodextrin (3) as well as mono(6-thy-6-deoxy)-beta-cyclodextrin (4), were selected as molecular receptors to investigate their conformation and inclusion complexation behaviors with some chiral molecules, that is, (+)-camphor, (-) camphor, (+)-borneol, and (-)-borneol, by spectrophotometric and microcalorimetric titrations in aqueous phosphate buffer solution (pH 7.2) at 298.15 K. Circular dichroism and NMR studies demonstrated that these nucleobase modified beta-cyclodextrins adopted a co-inclusion mode upon complexation with guest molecules; that is, the originally self-included nucleobase substituents of the host did not move out from the beta-cyclodextrin cavity, but coexisted with guest molecule in the beta-cyclodextrin cavity upon inclusion complexation. Significantly, these nucleobase-modified beta-cyclodextrins efficiently enhanced the molecular binding ability and the chiral recognition ability of native beta cyclodextrin, displaying enantioselectivity up to 3.7 for (+)-camphor/(-)-camphor pair by 2 and 3.5 for (-)-borneol/(+)-borneol pair by 3. The enhanced molecular/chiral recognition abilities of 2-4 toward (+/-)-camphor were mainly attributed to the increased entropic gains due to the extensive desolvation effects, while the favorable enthalpic gains originating from the good size-fit relationship as well as the hydrogen bond interactions between host and guest result in the enhanced molecular/chiral recognition abilities of 2-4 toward (+/-) borneol. PMID- 17914792 TI - Interference of ascorbic acid in the sensitive detection of dopamine by a nonoxidative sensing approach. AB - The electrochemistry of a poly(anilineboronic acid)/carbon nanotube composite was studied in the presence of dopamine and ascorbic acid. To understand the binding affinity of dopamine and ascorbic acid to the boronic acid functional groups in the composite, the association constants between the diol groups in dopamine and ascorbic acid and the boronic acid were experimentally determined using a fluorescence-based binding assay. The results demonstrate that ascorbic acid could severely interfere with the detection of dopamine in nonoxidative boronic acid-binding approaches: Ascorbic acid was able to electrocatalytically reduce the fully oxidized polyaniline backbone during the electrochemical oxidation process; similarly to dopamine, ascorbic acid was also able to bind to the boronic acid groups through its planar diol group even though the binding affinity is much lower. The examination of the dopamine transduction mechanism and ascorbic acid interference mechanism in this nonoxidative approach will benefit the design of future boronic acid-based sensors. PMID- 17914793 TI - Theoretical and experimental studies of enflurane. Infrared spectra in solution, in low-temperature argon matrix and blue shifts resulting from dimerization. AB - Theoretical studies are performed on enflurane (CHFCl-CF(2)-O-CHF(2)) to investigate the conformational properties and vibrational spectra. Calculations are carried out at the B3LYP/6-31G(d) level along with a natural bond orbital (NBO) analysis. Experimental infrared spectra are investigated in carbon tetrachloride solution at room temperature and in argon matrix at 12 K. In agreement with previously reported data (Pfeiffer, A.; Mack, H.-G.; Oberhammer, H. J. Am. Chem. Soc. 1998, 120, 6384), it is shown that the four most stable conformers possess a trans configuration of the C-C-O-C skeleton and a gauche orientation of the CHF(2) group (with respect to the central C-O bond). These conformations are favored by electrostatic interaction between the H atom of the CHF(2) group and the F atoms of the central CF(2) group. Hyperconjugation effects from the O lone pairs to the antibonding orbitals of the neighboring C-H and C-F bonds also contribute to the stability of the four conformers. The vibrational frequencies, infrared intensities, and potential energy distributions are calculated at the same level of theory for the most stable conformers. On the basis of the theoretical results, these conformers are identified in an argon matrix. The influence of the concentration on the nu(CH) vibrations suggests the formations of higher aggregates in solution. Theoretical calculations are carried out on the enflurane dimer. The results show that the dimer is formed between two enflurane conformers having the largest stability. The dimer has an asymmetric cyclic structure, the two enflurane molecules being held together by two nonequivalent C-H...F hydrogen bonds, the C-H bond of the CHFCl group acting as a proton donor, and one of the F atoms of the CHF(2) groups acting as a proton acceptor. The theory predicts a contraction of 0.0014-0.0025 A of the two CH bonds involved in the interaction along with a blue shift of 30-38 cm(-1) of the corresponding nu(C-H) bands, in good agreement with the blue shifts of 35-39 cm( 1) observed in an argon matrix. PMID- 17914794 TI - Promoting photochromism on flavylium derived 2-hydroxychalcones in aqueous solutions by addition of CTAB micelles. AB - A strategy to obtain photochromism from the network of chemical reactions originated by flavylium compounds in solution is described. This strategy is particularly useful for flavylium salts bearing amino groups which give rise to a variety of beautiful colors but lack photochemistry in water. The trans-chalcone of 7-(N,N-diethylamino)-4'-hydroxyflavylium interacts strongly with CTAB micelles defining a yellow dark state. Upon irradiation, the system switches to a pink-red state emerging from the flavylium cation that is formed inside the micelle and ejected to the bulk aqueous phase. The photochemical product reverts back to the trans-chalcone adduct with the micelle in the dark. The thermodynamics as well as the kinetics of the photochromic system were studied in detail. The best color contrast is obtained at pH = 4.25 with Phi = 0.001 and a recovery lifetime of approximately 3 h. This photochromic system works with no need of changing the pH, which constitutes an important improvement over previously described systems dependent on pH jumps. PMID- 17914796 TI - Tuning the molecular properties of polybenzimidazole by copolymerization. AB - In the present work, a series of novel random polybenzimidazole (PBI) copolymers consisting of m- and p-phenylene linkages are synthesized from various stoichiometric mixtures of isophthalic acid (IPA) and terephthalic acid (TPA) with 3,3',4,4'-tetraaminobiphenyl (TAB) by solution copolycondensation in polyphosphoric acid (PPA). The resulting copolymers are characterized by different techniques to obtain their molecular properties parameters. The monomer concentration in the polymerization plays an important role in controlling the molecular weight of the polymer. Surprisingly, a simple change in the dicarboxylic acid architecture from meta (IPA) to para (TPA) increases the molecular weight of the copolymers, which is maximum for the para homopolymer. The low solubility of TPA in PPA is found to be the dominating factor for obtaining the higher molecular weight polymer in the case of the para structure. FT-IR study shows that the introduction of the para structure enhances the conjugation along the polymer chain. The positive deviation of the copolymer composition from the feed ratio is due to the higher reactivity ratio of TPA than IPA, which is obtained from proton NMR studies. The incorporation of the para structure in the chain enhances the thermal stability of the polymers. The para homopolymer shows 59 degrees C lower glass transition temperature compare to the meta homopolymer indicating enhancement of the flexibility of the polymer chain due the introduction of the p-phenylene linkage in the backbone. The T(g) of the copolymers shows both positive and negative deviation from the expected T(g) calculated by the Fox equation. The enhanced conjugation of the polymer chains also influences the photophysical properties of the polymers in solution. All the PBI polymers exhibit strong fluorescence in dimethylacetamide solution. As expected, that all the polymers are amorphous in nature reveals that the copolymerization does not influence the packing characteristics of the PBI chains. PMID- 17914795 TI - Ca2+ selectivity of the sarcoplasmic reticulum Ca2+-ATPase at the enzyme-water interface and in the Ca2+ entrance channel. AB - The sarcoplasmic reticulum (SR) Ca(2+)-ATPase, a P-type transmembrane protein, can transport Ca(2+) from the cytoplasmic to the luminal side over other cations specifically. The proposed Ca(2+) entrance channel, composed of the main-chain carbonyl oxygen and side-chain carboxyl oxygen atoms of the amino acids, opens on the enzyme surface, just above the biphospholipid layer membrane-water interface, where Trp residues are frequently found. In this work, the physicochemical nature of Ca(2+) selectivity over Mg(2+) on the surface of the SR Ca(2+)-ATPase has been investigated using the density functional theory (DFT) method. The selection process can be regarded as the first step of the specificity of the enzyme to transport Ca(2+). Subsequently, the specificity of the entrance channel to conduct Ca(2+) over other cations has also been explored. As revealed by thermodynamic analyses, either the aromatic or the aliphatic amino acid residues distributed on the surface of Ca(2+)-ATPase have a bigger affinity to Mg(2+) than to Ca(2+), resulting in a concentration decrease of free Mg(2+) in the local region. Thus, Ca(2+) can transport into the Ca(2+)-entrance channel more easily. Whereafter, for a small quantity of Mg(2+) entering this channel accompanying the Ca(2+) current, the strong electrostatic interactions between Mg(2+) and the ligands will limit the activity of this metal ion, which facilitates the weakly bonded Ca(2+) passing through the channel at a relatively high rate, as suggested by the "sticky-pore" hypothesis. Furthermore, the corresponding theoretical investigations have demonstrated that the increase of the ligand electronegativity can enhance their discrimination between these two cations effectively. PMID- 17914797 TI - Kinetic coupled with UV spectral evidence for near-irreversible nonionic micellar binding of N-benzylphthalimide under the typical reaction conditions: an observation against a major assumption of the pseudophase micellar model. AB - Pseudo-first-order rate constants (k(obs)) for alkaline hydrolysis of N benzylphthalimide (1) show a nonlinear decrease with the increase in [C(m)E(n)]T (total concentration of Brij 58, m = 16, n = 20 and Brij 56, m = 16, n = 10) at constant [CH(3)CN] and [NaOH]. These nonionic micellar effects, within the certain typical reaction conditions, have been explained in terms of the pseudophase micellar (PM) model. The values of micellar binding constants (KS) of 1 are 1.04 x 10(3) M(-1) (at 1.0 x 10(-3) M NaOH) and 1.08 x 10(3) M(-1) (at 2.0 x 10(-3) M NaOH) for C(16)E(20) as well as 600 M(-1) (at 7.6 x 10(-4) M NaOH) and 670 M(-1) (at 1.0 x 10(-3) M NaOH) for C(16)E(10) micelles. The pseudo-first order rate constants (kM) for hydrolysis of 1 in C(16)E(20) micellar pseudophase are approximately 90-fold smaller than those (kW) in water phase. The values of kM for hydrolysis of 1 in C(16)E(10) micelles are almost zero. Kinetic coupled with UV spectral data reveals significant irreversible nonionic micellar binding of 1 molecules in the micellar environment of nearly zero hydroxide ion concentration at >or=0.14 M C(16)E(20) and 1.0 x 10(-3) M NaOH while such observations could not be detected at or=3 x 10(-3) M C(16)E(10) and 7.6 x 10(-4) M NaOH, while the rate of hydrolysis of 1 is completely ceased at >or=0.05 M C(16)E(10) and 7.6 x 10(-4) M NaOH. The rate of hydrolysis of 1 at 5.0 x 10(-2) and 8.8 x 10(-2) M C(16)E(10) and 1.0 x 10(-3) M NaOH reveals the formation of presumably phthalic anhydride, whereas such observation was not observed in the C(16)E(20) micellar system under similar experimental conditions. PMID- 17914798 TI - Synthesis of lanthanum-doped MCM-48 molecular sieves and its catalytic performance for the oxidation of styrene. AB - Lanthanum-doped MCM-48 molecular sieves with different La contents were synthesized hydrothermally and characterized by X-ray diffraction (XRD), nitrogen sorption, transmission electron microscopy (TEM), UV-visible spectroscopy, and Fourier transform infrared (FT-IR) spectroscopy. The results show that the majority of La cations have been incorporated into the framework of MCM-48 molecular sieves. When the molar ratio of La/Si is >0.039 in the sample, some of lanthanum species exist in the extraframework. Compared with pure silicate MCM 48, lanthanum-doped MCM-48 samples show the medium strong acidity that is due to the incorporation of La in the framework of silica. In the oxidation of styrene with H(2)O(2) as the oxidant over the lanthanum-doped MCM-48 catalysts, benzaldehyde is the main product with a small amount of styrene oxide. The La content in the catalysts, reaction temperature, reaction time, and solvent affect greatly the catalytic oxidation of styrene. The conversion of styrene and the selectivity to styrene oxide increase noticeably when a small amount of NaOH aqueous solution is added into the reaction mixture. Ln-doped MCM-48 catalysts with 14 kinds of rare earth elements were synthesized hydrothermally and evaluated for the oxidation of styrene. The results show that their catalytic performance is tremendously different and depends on the nature of rare earth elements doped in the MCM-48 mesoporous materials. PMID- 17914799 TI - Imidazolium-based ionic liquids formed with dicyanamide anion: influence of cationic structure on ionic conductivity. AB - Ionic liquids composed of dicyanamide anion and various imidazolium-based cations were prepared, and the influence of structural variations such as substituting a hydrogen at 2-position and changing the sort of alkyl group at 1-position of imidazolium cations on their thermal behavior, density, solvatochromic effects, viscosity, ionic conductivity, and surface tension was characterized. The substitution of the 2-hydrogen for methyl group or N-methylimidazole decreases the fluidity and ionic conductivity, mainly caused by the increased cohesive energy associated with the increasing cation size. Chain branching at 1-position also gives rise to the pronounced depression of the fluidity and ionic conductivity, presumably as a consequence of the increased pi-pi interactions between imidazolium rings. We found that the surface tension of the present ionic liquids is in inverse proportion to the molar concentration, which can be originally rationalized on the basis of the hole theory. PMID- 17914800 TI - Hydration and dynamic behavior of cyclodextrins in aqueous solution. AB - The hydration state and dynamics of plain and chemically modified cyclodextrins (CDs) in aqueous solution were investigated by using dielectric relaxation measurements at 25 degrees C over a wide frequency range up to 20 GHz, which is the relaxation frequency of pure liquid water molecules. The obtained dielectric relaxation spectra were decomposed into two major and one minor relaxation modes with relaxation times of approximately 8.3, 20-25, and 1000-2500 ps, respectively, depending on the CD species. The two major modes, fast and medium, were attributed to a rotational relaxation process of water molecules belonging to the bulk (free) state and an exchange of water molecules hydrated to CDs owing to hydrogen bond formation. The hydration numbers of the CDs strongly depend on the number of hydroxy (OH) groups controlled by chemical modification such as methylation. Increasing the number of methoxy or 2-hydroxypropoxy groups increases the hydration number of CD molecules, and results in higher solubilities of the chemically modified CDs than those of the plain CDs. The minor, slow mode was assigned to overall rotational relaxation for CDs with finite permanent dipole moments, which also depends on the number of OH groups. PMID- 17914801 TI - Preferential hydroxylation over epoxidation catalysis by a horseradish peroxidase mutant: a cytochrome P450 mimic. AB - Density functional theory calculations are presented on the catalytic properties of a horseradish peroxidase mutant whereby the axial nitrogen atom is replaced by phosphorus. This mutant has never been studied experimentally and only one theoretical report on this system is known (de Visser, S. P. J. Phys. Chem. B 2006, 110, 20759-20761). Thus, a one-atom substitution in horseradish peroxidase changes the properties of the catalytic center of the enzyme to more cytochrome P450-type qualities. In particular, the phosphorus-substituted horseradish peroxidase mutant reacts with substrates via a unique reactivity pattern, whereby alkanes are regioselectively hydroxylated even in the presence of a double bond. Reaction barriers of propene epoxidation and hydroxylation are almost identical to ones observed for a cytochrome P450 catalyst and significantly higher than those obtained for a horseradish peroxidase catalyst. It is shown that the regioselectivity difference is entropy and thermally driven and that the electron transfer processes that occur during the reaction mechanism follow cytochrome P450-type patterns in the hydroxylation reaction. PMID- 17914803 TI - Solid-state 13C NMR studies on organic-inorganic hybrid zeolites. AB - A novel type of organic-inorganic hybrid zeolite with organic lattice (ZOL) is studied in detail by solid-state (13)C magic angle spinning nuclear magnetic resonance (MAS NMR). The (13)C MAS NMR measurements employing several pulse sequences quantitatively demonstrate that methylene groups are really incorporated in the framework, although they are partially cleaved into methyl groups. The organic species in ZOL materials are open for adsorbates, which is evidenced by the (13)C MAS NMR measurements for an n-hexane-adsorbing ZOL material. This finding strongly suggests that organic moieties are incorporated as a zeolite framework, indicating that ZOL is not a physical mixture of a carbon containing amorphous aggregate and a conventional zeolite but a true organic inorganic hybrid zeolite. PMID- 17914802 TI - Dynamic control of protein folding pathway with a polymer of tunable hydrophobicity. AB - While the knowledge of protein folding in a dilute solution is now well-advanced, little is known of the influence of surrounding conditions on the folding kinetics, in particular when the protein is in a dynamically responsive environment. Here we report a new procedure to control the pathways of protein folding by using a thermally responsive polymer that varies its hydrophobicity concomitant with the protein structural changes. The advantages of folding in a dynamic environment have been demonstrated first by Langevin dynamics simulations on the basis of coarse-grained models for both the protein and polymer and then by experiments for lysozyme refolding in the presence of poly(N isopropylacrylamide-co-N-tert-butylacrylamide), a thermal responsive polymer that varies its hydrophobicity in response to temperature. The simulation suggests that decreasing the polymer hydrophobicity during the folding process may result in an optimized free-energy landscape that enhances both the folding yield and kinetics. The experiments affirm that an optimal folding condition can be identified when structural transitions of the protein collaborate with the polymer hydrophobicity tuned by variation of temperature. PMID- 17914804 TI - Calcium borohydride for hydrogen storage: catalysis and reversibility. AB - We demonstrate a new solid-state synthesis route to prepare calcium borohydride, Ca(BH4)2, by reacting a ball-milled mixture of CaB(6) and CaH(2) in a molar ratio of 1:2 at 700 bar of H2 pressure and 400-440 degrees C. Moreover, doping with catalysts was found to be crucial to enhance reaction kinetics. Thermogravimetric analysis and differential scanning calorimetry revealed a reversible low temperature to high-temperature endothermic phase transition at 140 degrees C and another endothermic phase transition at 350-390 degrees C associated with hydrogen release upon formation of CaB(6) and CaH(2), as was evident from X-ray diffraction analysis. Thus, since Ca(BH(4))(2) here is shown to be prepared from its anticipated decomposition products, the conclusion is that it has potential to be utilized as a reversible hydrogen storage material. The theoretical reversible capacity was 9.6 wt % hydrogen. PMID- 17914805 TI - Reinvestigation of carrier transport properties in liquid crystalline 2 phenylbenzothiazole derivatives. AB - We have reinvestigated the charge carrier transport properties in a liquid crystal of 2-(4'-heptyloxyphenyl)-6-dodecylthiobenzothiazole (7O-PBT-S12), for which the electronic conduction was first established in rodlike liquid crystals and for which the highest hole mobility in the smectic A (SmA) phase ever achieved was reported. We found that 7O-PBT-S12 exhibited three crystal phases, one of which appeared in a limited temperature range of 10 degrees just below the phase transition temperature from the SmA phase. In this crystal phase, nondispersive transient photohole currents were observed in time-of-flight experiments, and its hole mobility was determined to be 8 x 10(-3) cm(2)/Vs, slightly higher than that reported previously in the SmA phase. For the SmA phase, however, the hole mobility was 1 x 10(-4) cm(2)/Vs. Furthermore, we established the electron transport in the SmA phase of purified 7O-PBT-S12, whose mobility was the same as the hole mobility in that phase. In order to confirm generality of the new findings in 7O-PBT-S12, we investigated the carrier transport properties of its derivative having a short hydrocarbon chain, 2-(4' heptyloxyphenyl)-6-butylthiobenzothiazole (7O-PBT-S4), and obtained comparable results. The present results correct a mistake in the previous report and give an idea of what a typical mobility in the SmA phase is. On the basis of these results, we discuss what determines the charge carrier mobility in smectic mesophases. PMID- 17914806 TI - Magnetic circular dichroism of porphyrins containing M = Ca, Ni, and Zn. A computational study based on time-dependent density functional theory. AB - A theoretical study is presented on the magnetic circular dichroism (MCD) exhibited by the porphyrin complexes MP (M = Mg,Ni,Zn), MTPP (M = Mg,Ni,Zn), and NiOEP, where P = porphyrin, TPP = tetraphenylporphyrin, and OEP = octaethylporphyrin. The study makes use of a newly implemented method for the calculation of A and B terms from the theory of MCD and is based on time dependent density functional theory (TD-DFT). It is shown that the MCD spectrum is dominated by a single positive A term in the Q-band region in agreement with experiment where available. The band can be fully explained as the first transition in Gouterman's four-orbital model for the type of porphyrins studied here. For the Soret band, the experimental MCD spectrum appears as a single positive A term. This is also what is found computationally for NiP and NiTPP, where the second transition in Gouterman's four-orbital model give rise to a positive A term. However, for the remaining systems, the simulated MCD spectrum is actually due to two B terms that have the appearance of one positive pseudo A term. The two B terms appear because the second Gouterman state is coupled strongly to a second excited state (b(2u) --> 2e(g)) of nearly the same energy by the external magnetic field. PMID- 17914807 TI - Oxidative addition of small molecules to a dinuclear Au(I) amidinate complex, Au2[(2,6-Me2Ph)2N2CH]2. Syntheses and characterization of Au(II) amidinate complexes including one which possesses Au(II)-oxygen bonds. AB - The dinuclear Au(I) amidinate complex Au2(2,6-Me2Ph-form)2 (1) is isolated in quantitative yield by the reaction of (THT)AuCl and the potassium salt of 2,6 Me2Ph-form in a 1:1 stoichiometric ratio. Various reagents such as Cl2, Br2, I2, CH3I, and benzoyl peroxide add to the dinuclear Au(I)amidinate complex Au2(2,6 Me2Ph-form)2 to form oxidative-addition Au(II) metal-metal-bonded complexes 2, 3, 4, 5, and 6. The Au(II) amidinate complexes are stable as solids at room temperature. The structures of the dinuclear Au2(2,6-Me2Ph-form)2 and the Au(II) oxidative-addition products Au2(2,6-Me2Ph-form)2X2, X=Cl, Br, I, are reported. Crystalline products with an equal amount of oxidized and unoxidized complexes in the same unit cell, [Au2(2,6-Me2Ph-form)2X2][Au2(2,6-Me2Ph-form)2], X=Cl, 2m, or Br, 3m, are isolated and their structures are presented. The structure of [Au2(2,6-Me2Ph-form)2X2][Au2(2,6-Me2Ph-form)2], X=Cl has a Au(II)-Au(II) distance slightly longer, 0.05A, than that observed in the fully oxidized product Au2(2,6 Me2-form)2Cl2, 2. The gold-gold distance in the dinuclear complex decreases upon oxidative addition with halogens from 2.7 to 2.5 A, similar to observations made with the Au(I) dithiolates and ylides. The oxidative addition of benzoyl peroxide leads to the isolation of the first stable dinuclear Au(II) nitrogen complex possessing Au-O bonds, Au2(2,6-Me2Ph-form)2(PhCOO)2, 6, with the shortest Au-Au distance known for Au(II) amidinate complexes, 2.48 A. The structure consists of unidentate benzoate units linked through oxygen to the Au(II) centers. The replacement of the bromide in 3 by chloride, and the benzoate groups in 6 by chloride or bromide also occurs readily. The unit cell dimensions are, for 1, a=7.354(6) A, b=9.661(7) A, c=11.421(10) A, alpha=81.74(5) degrees, beta=71.23(5) degrees, and gamma=86.07(9) degrees (space group P, Z=1), for 2.1.5C6H12, a=11.012(2) A, b=18.464(4) A, c=19.467(4) A, alpha=90 degrees, beta=94.86(3) degrees, and gamma=90 degrees (space group P21/c, Z=4), for 2m.ClCH2CH2Cl, a=16.597(3) A, b=10.606(2) A, c=19.809(3) A, alpha=90 degrees, beta=94.155(6) degrees, and gamma=90 degrees (space group P21/n, Z=2), for 3m, a=16.967(3) A, b=10.783(2) A, c=20.060(4) A, alpha=90 degrees, beta=93.77(3) degrees, and gamma=90 degrees (space group P21/n, Z=2), for 4.THF, a=8.0611(12) A, b=10.956(16) A, c=11.352(17) A, alpha=84.815(2) degrees, beta=78.352(2) degrees, and gamma=88.577(2) degrees (space group P, Z=1), for 5, a=16.688 A, b=10.672(4) A, c=19.953(7) A, alpha=90.00 (6) degrees, beta=94.565(7) degrees, and gamma=90.00 degrees (space group P21/n, Z=4), for 6.0.5C7H8, a=11.160(3) A, b=12.112(3) A, c=12.364(3) A, alpha=115.168(4) degrees, beta=161.112(4) degrees, and gamma=106.253(5) degrees (space group P, Z=1). PMID- 17914808 TI - 1,2-hydroxypyridonates as contrast agents for magnetic resonance imaging: TREN 1,2-HOPO. AB - 1,2-Hydroxypyridinones (1,2-HOPO) form very stable lanthanide complexes that may be useful as contrast agents for magnetic resonance imaging (MRI). X-ray diffraction of single crystals established that the solid-state structures of the Eu(III) and the previously reported [Inorg. Chem. 2004, 43, 5452] Gd(III) complex are identical. The recently discovered sensitizing properties of 1,2-HOPO chelates for Eu(III) luminescence [J. Am. Chem. Soc. 2006, 128, 10 067] allow for direct measurement of the number of water molecules coordinated to the metal center. Fluorescence measurements of the Eu(III) complex corroborate that, in solution, two water molecules coordinate the lanthanide (q = 2) as proposed from the analysis of NMRD profiles. In addition, fluorescence measurements have verified the anion binding interactions of lanthanide TREN-1,2-HOPO complexes in solution, studied by relaxivity, revealing only very weak oxalate binding (KA = 82.7 +/- 6.5 M-1). Solution thermodynamic studies of the metal complex and free ligand have been carried out using potentiometry, spectrophotometry, and fluorescence spectroscopy. The metal ion selectivity of TREN-1,2-HOPO supports the feasibility of using 1,2-HOPO ligands for selective lanthanide binding [pGd = 19.3 (2), pZn = 15.2 (2), pCa = 8.8 (3)]. PMID- 17914809 TI - Syntheses, structures, magnetism, and optical properties of lutetium-based interlanthanide selenides. AB - Ln3LuSe6 (Ln = La, Ce), beta-LnLuSe3 (Ln = Pr, Nd), and LnxLu4-xSe6 (Ln = Sm, Gd; x = 1.82, 1.87) have been synthesized using a Sb2Se3 flux at 1000 degrees C. Ln3LuSe6 (Ln = La, Ce) adopts the U3ScS6-type three-dimensional structure, which is constructed from two-dimensional 2(infinity)[Ln3Se6](3-) slabs with the gaps between these slabs being filled by octahedrally coordinated Lu(3+) ions. The series of beta-LnLuSe3 (Ln = Pr, Nd) are isotypic with UFeS3. Their structures include layers formed from LuSe6 octahedra that are separated by eight-coordinate Ln(3+) (Ln = Pr, Nd) ions in bicapped trigonal prismatic environments. Sm1.82Lu2.18Se6 and Gd1.87Lu2.13Se6 crystallize in the disordered F-Ln2S3 type structure with the eight-coordinate bicapped trigonal prismatic Ln(1) ions residing in the one-dimensional channels formed by three different double chains via edge- and corner-sharing. These double chains are constructed from Ln(2)Se7 monocapped trigonal prisms, Ln(3)Se6 octahedra, and Ln(4)S6 octahedra, respectively. The magnetic susceptibilities of beta-PrLuSe3 and beta-NdLuSe3 follow the Curie-Weiss law. Sm1.82Lu2.18Se6 shows van Vleck paramagnetism. Magnetic susceptibility measurements show that Gd1.87Lu2.13Se6 undergoes an antiferromagnetic transition around 4 K. Ce3LuSe6 exhibits soft ferromagnetism below 5 K. The optical band gaps for La3LuSe6, Ce3LuSe6, beta-PrLuSe3, beta NdLuSe3, Sm1.82Lu2.18Se6, and Gd1.87Lu2.13Se6 are 1.26, 1.10, 1.56, 1.61, 1.51, and 1.56 eV, respectively. PMID- 17914810 TI - Volatile magnesium octahydrotriborate complexes as potential CVD precursors to MgB2. Synthesis and characterization of Mg(B3H8)2 and its etherates. AB - The solid-state reaction of MgBr2 and NaB3H8 at 20 degrees C, followed by sublimation at 80 degrees C and 0.05 Torr, affords Mg(B3H8)2 as a white solid. Similar reactions with MgBr2(Et2O) and MgBr2(Me2O)1.5 afford the crystalline ether adducts Mg(B3H8)2(Et2O)2 and Mg(B3H8)2(Me2O)2, respectively. In contrast, reactions of MgBr2 with NaB3H8, the presence of excess solvent result in the formation of nonvolatile, probably ionic, magnesium compounds of the type [MgLx][B3H8]2. The adducts Mg(B3H8)2(Et2O)2 and Mg(B3H8)2(Me2O)2 are the first crystallographically characterized magnesium complexes of the B3H8- ligand; in both structures, the magnesium center adopts a distorted cis-octahedral geometry with two bidentate B3H8 groups and two Et2O ligands. Owing to their volatility, Mg(B3H8)2(Et2O)2 and Mg(B3H8)2(Me2O)2 are potential precursors for the deposition of MgB2 thin films, although preliminary efforts to employ them as chemical vapor deposition sources produce boron-rich MgBx films instead, with x approximately 7. Finally, the synthesis and structure of Cp2Mg(thf) are described: this mono-thf adduct of Cp2Mg bears two eta5-Cp groups, unlike other Lewis base adducts of Cp2Mg, which contain one eta5-Cp group and one eta1- or eta2-Cp group. PMID- 17914811 TI - Reactions of fac-[Re(CO)3(H2O)3]+ with nucleoside diphosphates and thiamine diphosphate in aqueous solution investigated by multinuclear NMR spectroscopy. AB - Products formed between monoester diphosphates (MDPs) and fac-[Re(CO)3(H2O)3]OTf at pH 3.6 were examined. Such adducts of the fac-[Re(CO)3]+ moiety have an uncommon combination of properties for an "inert" metal center in that sharp NMR signals can be observed, yet the products are equilibrating at rates allowing NMR EXSY cross-peaks to be observed. Thiamine diphosphate (TDP) and uridine 5' diphosphate (5'-UDP) form 1:1 bidentate {Palpha,Pbeta} chelates, in which the MDP binds Re(I) via Palpha and Pbeta phosphate groups. Asymmetric centers are created at Re(I) (RRe/SRe) and Palpha (Delta/Lambda), leading to four diastereomers. The two mirror pairs of diastereomers (RReDelta/SReLambda) and (RReLambda/SReDelta) for TDP (no ribose) and for all four diastereomers (RReDelta, RReLambda, SReDelta, SReLambda) for 5'-UDP (asymmetric ribose) gave two and four sets of NMR signals for the bound MDP, respectively. 31Palpha-31Palpha EXSY cross-peaks indicate that the fac-[Re(CO)3(H2O)({Palpha,Pbeta}MDP)]- isomers interchange slowly on the NMR time scale, with an average k approximately equal to 0.8 s(-1) at 32 degrees C; the EXSY cross-peaks could arise from chirality changes at only Re(I) or at only Palpha. Guanosine 5'-diphosphate (5'-GDP), with a ribose moiety and a Re(I)-binding base, formed both possible diastereomers (RRe and SRe) of the fac-[Re(CO)3(H2O)({N7,Pbeta}GDP)]- macrochelate, with one slightly more abundant diastereomer suggested to be RRe by Mn2+ ion 1H NMR signal line-broadening combined with distances from molecular models. Interchange of the diastereomers requires that the coordination site of either N7 or Pbeta move to the H2O site. 31Palpha-31Palpha EXSY cross-peaks indicate a k approximately equal to 0.5 s(-1) at 32 degrees C for RRe-to-SRe interchange. The similarity of the rate constants for interchange of fac-[Re(CO)3(H2O)({Palpha,Pbeta}MDP)]- and fac [Re(CO)3(H2O)({N7,Pbeta}GDP)]- adducts suggest strongly that interchange of Pbeta and H2O coordination positions accounts for the EXSY cross-peaks present in the spectra of all adducts. PMID- 17914812 TI - Structure and magnetic ordering of KxH1-xNi(OH2)4[Ru2(CO3)4].zH2O. AB - K(x)H(1-x)Ni(OH2)4[Ru2(CO3)4].zH2O is a ferrimagnet (Tc = 4.3 K) formed from the reaction of K3[Ru(II/III)2(CO3)4] and Ni(II) in water. It possesses a new 3-D network structural motif composed of linked chains and mu3-CO3 linkages to both Ru and Ni sites. Each Ni(II) bonds to four oxygens and to two [Ru2(CO3)4]3- moieties in a cis manner, and four mu3-CO3 groups from each [Ru2(CO3)4](3-) have two oxygens bonding to the Ru2 moiety, forming the typical paddle-wheel core, and trans pairs of the third CO32- oxygen axially bonded to either another Ru2 or Ni(II). PMID- 17914813 TI - Trapping of anionic organic radicals by (TpMe2)2Ln (Ln = Sm, Eu). AB - Stoichiometric reaction of [ Sm(Tp(Me2))2 ], 1, with a variety of reducible ketone- and quinone-type substrates gave thermally stable, isolable radical anions/ketyls in moderate to good yields. Thus reaction with benzophenone gave [Sm(Tp(Me2))2(OCPh2)], 2, with fluorenone [Sm(Tp(Me2))2(eta1-OC13H8)], 3, and di tert-butylparaquinone [Sm(Tp(Me2))2(eta1-OC6H2(tBu)2O)], 4, each of which was structurally characterized. In the case of the less-hindered benzoquinone, an unimetallic semiquinone [Sm(Tp(Me2))2(OC6H4O)], 5, could be isolated, although it was unstable with respect to formation of the dimetallic complex [Sm(Tp(Me2))2]2(mu-OC6H4O), 6. Compound 6 was structurally characterized, as was its anthraquinone analogue [Sm(Tp(Me2))2]2(mu-OC14H8O), 7. When the analogous reaction was carried out between the less-reducing [Eu(Tp(Me2))2] and benzoquinone, only the europium analogue of the semiquinone 5, [Eu(Tp(Me2))2(OC6H4O)], 8, could be isolated. The use of the sterically hindered 3,5-di-tert-butyl-o-benzoquinone allowed isolation of [Sm(Tp(Me2))2(DTBSQ)], 9. PMID- 17914814 TI - Aerosol assisted chemical vapor deposition of In2O3 films from Me3In and donor functionalized alcohols. AB - The reaction of Me3In and ROH (R = CH2CH2NMe2, CH(CH3)CH2NMe2, C(CH3)2CH2OMe, CH2CH2OMe) in toluene under aerosol assisted chemical vapor deposition (AACVD) conditions leads to the production of indium oxide thin films on glass. The indium oxide films were deposited at 550 degrees C and analyzed by scanning electron microscopy (SEM), X-ray powder diffraction, wavelength dispersive analysis of X-rays (WDX), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopy. This CVD technique offers a rapid, convenient route to In2O3, which presumably involves the in situ formation of dimethylindium alkoxides, of the type [Me2InOR]2. In order to identify compounds present in the aerosol mist, the solution-phase reaction between Me3In and ROH (R = CH2CH2NMe2, C(CH3)2CH2OMe, CH(CH3)CH2NMe2, CH(CH2NMe2)2) at room temperature in toluene was carried out. Dimeric indium alkoxides, of the type [Me2In(OR)]2, were isolated, and their structures were determined by X-ray crystallography. PMID- 17914815 TI - New polypyridine ligands functionalized with redox-active Fe(II) organometallic fragments. AB - We report in this Communication the isolation and characterization, including structure determinations, of 2,2',6',2"-terpyridine (2) and 2,2'-bipyridine (3) ligands bearing two redox-active "(eta2-dppe)(eta5-C5Me5)FeC[triple bond]C-" moieties grafted to the 5 and 5" positions of terpy or to the 5 and 5' positions of bipy. These "metalloligands" have been complexed with Ru(II) and Mo(0), providing new heterotrinuclear complexes displaying intense absorptions around 700 and 600 nm, respectively, for the Fe2Ru/terpy and Fe2Mo/bipy species. In both cases, the Fe(II)/Fe(III) oxidation potentials of the free ligands became more positive by more than 50 mV upon complexation. PMID- 17914816 TI - Benzimidazole and related ligands for Cu-catalyzed azide-alkyne cycloaddition. AB - Tris(2-benzimidazolylmethyl)amines have been found to be superior accelerating ligands for the copper(I)-catalyzed azide-alkyne cycloaddition reaction. Candidates bearing different benzimidazole N-substituents as well as benzothiazole and pyridyl ligand arms were evaluated by absolute rate measurements under relatively dilute conditions by aliquot quenching kinetics and by relative rate measurements under concentrated conditions by reaction calorimetry. Benzimidazole-based ligands with pendant alkylcarboxylate arms proved to be advantageous in the latter case. The catalyst system was shown to involve more than one active species, providing a complex response to changes in pH and buffer salts and the persistence of high catalytic rate in the presence of high concentrations of coordinating ligands. The water-soluble ligand (BimC4A)3 was found to be especially convenient for the rapid and high-yielding synthesis of several functionalized triazoles with 0.01-0.5 mol % Cu. PMID- 17914817 TI - Ligand-accelerated Cu-catalyzed azide-alkyne cycloaddition: a mechanistic report. AB - The experimental rate law for the Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction was found to vary in complex ways with concentration, the presence of chloride ion, and the presence of accelerating ligands. Several examples of discontinuous ("threshold behavior") kinetics were observed, along with a decidedly nonlinear correlation of electronic substituent parameter with the rate of CuAAC reaction with p-substituted arylazides. The previously observed tendency of the CuAAC reaction to provide ditriazoles from a conformationally constrained 1,3-diazide was found to be affected by a class of polybenzimidazole ligands introduced in the accompanying article. Various lines of evidence suggest that the standard tris(triazolylmethyl)amine ligand binds less strongly to Cu(I) than its benzimidazole analogues. On the basis of these observations, it is proposed that (a) a central nitrogen donor provides electron density at Cu(I) that assists the cycloaddition reaction, (b) the three-armed motif bearing relatively weakly coordinating heterocyclic ligands serves to bind the metal with sufficient strength while providing access to necessary coordination site(s), (c) at least two active catalysts or mechanisms are operative under the conditions studied, and (d) pendant acid or ester arms in the proper position can assist the reaction by speeding the protiolysis step that cleaves the Cu-C bond of a Cu.triazolyl intermediate. PMID- 17914819 TI - Rhodium-catalyzed carbonylation of spiropentanes. PMID- 17914820 TI - Electrophile-directed diastereoselective alkylation of prochiral enediolates. PMID- 17914818 TI - Anomeric reactivity-based one-pot synthesis of heparin-like oligosaccharides. AB - A highly efficient one-pot methodology is described for the synthesis of heparin and heparan sulfate oligosaccharides utilizing thioglycosides with well-defined reactivity as building blocks. L-Idopyranosyl and D-glucopyranosyl thioglycosides 5 and 10 were used as donors due to low reactivity of uronic acids as the glycosyl donors in the one-pot synthesis. The formation of uronic acids by a selective oxidation at C-6 was performed after assembly of the oligosaccharides. The efficiency of this programmable strategy with the flexibility for sulfate incorporation was demonstrated in the representative synthesis of disaccharides 17, 18, tetrasaccharide 23, and pentasaccharide 26. PMID- 17914821 TI - Organic-inorganic nanocomposites via directly grafting conjugated polymers onto quantum dots. AB - Nanocomposites of poly(3-hexylthiophene)-cadmium selenide (P3HT-CdSe) were synthesized by directly grafting vinyl-terminated P3HT onto [(4 bromophenyl)methyl]dioctylphosphine oxide (DOPO-Br)-functionalized CdSe quantum dot (QD) surfaces via a mild palladium-catalyzed Heck coupling, thereby dispensing with the need for ligand exchange chemistry. The resulting P3HT-CdSe nanocomposites possess a well-defined interface, thus significantly promoting the dispersion of CdSe within the P3HT matrix and facilitating the electronic interaction between these two components. The photophysical properties of nanocomposites were found to differ from the conventional composites in which P3HT and CdSe QDs were physically mixed. Solid-state emission spectra of nanocomposites suggested the charge transfer from P3HT to CdSe QDs, while the energy transfer from 3.5 nm CdSe QD to P3HT was implicated in the P3HT/CdSe composites. A faster decay in lifetime further confirmed the occurrence of charge transfer in P3HT-CdSe nanocomposites. PMID- 17914822 TI - An illustration of the limit of the metal organic framework's isoreticular principle using a semirigid tritopic linker obtained by "click" chemistry. PMID- 17914823 TI - Phosphine-catalyzed [4 + 2] annulation: synthesis of cyclohexenes. PMID- 17914824 TI - Unexpected coordination chemistry of bisphenanthroline complexes within hybrid materials: a mild way to Eu(2+) containing materials with bright yellow luminescence. PMID- 17914825 TI - Enantioselective iridium-catalyzed imine vinylation: optically enriched allylic amines via alkyne-imine reductive coupling mediated by hydrogen. PMID- 17914826 TI - Geminal tandem C-methylation in the discoid resistomycin pathway. PMID- 17914827 TI - Origin of regioselectivity in palladium-catalyzed cross-coupling reactions of polyhalogenated heterocycles. PMID- 17914828 TI - Protein prosthesis: 1,5-disubstituted[1,2,3]triazoles as cis-peptide bond surrogates. PMID- 17914833 TI - Synthesis, properties, and structures of benzo[1,2-b:4,5-b']bis[b]benzothiophene and benzo[1,2-b:4,5-b']bis[b]benzoselenophene. AB - Employing two consecutive cyclization reactions, benzo[1,2-b:4,5 b']bis[b]benzochalcogenophenes, which are pi-extended heteroarenes, were efficiently synthesized. Their electronic and crystal structures were elucidated on the basis of UV-vis spectra, electrochemical measurements, and X-ray structural analyses. PMID- 17914829 TI - The direction of glycan chain elongation by peptidoglycan glycosyltransferases. PMID- 17914834 TI - Protecting a squaraine near-IR dye through its incorporation in a slippage derived [2]rotaxane. AB - The photophysical properties and chemical stability of a squaraine derivative were enhanced after using Na+ ions to template a high-yield slippage synthesis of a [2]rotaxane from the dye and a molecular cage. PMID- 17914835 TI - Catalytic asymmetric synthesis of allylsilanes through rhodium/chiral diene catalyzed 1,4-addition of alkenyl[2-(hydroxymethyl)phenyl]dimethylsilanes. AB - A new synthetic method of chiral allylsilanes has been developed through a rhodium-catalyzed asymmetric 1,4-addition of alkenyl[2 (hydroxymethyl)phenyl]dimethylsilanes to beta-silyl alpha,beta-unsaturated ketones. By employing (S,S)-Ph-bod* as a ligand, a range of alkenyl nucleophiles have been installed to these substrates in high yield and enantiomeric excess. The resulting allylsilanes can be used for stereoselective intramolecular allylation reactions to control two contiguous tertiary and quaternary stereocenters. PMID- 17914836 TI - AuCl-catalyzed synthesis of benzyl-protected substituted phenols: a formal [3+3] approach. AB - A AuCl-catalyzed synthesis of highly substituted, benzyl-protected phenols is developed. This reaction unites enal/enones and benzyl allenyl ether in a [3+3] fashion in two steps, allowing flexibility in phenol synthesis and excellent control of substitution at the benzene ring. PMID- 17914837 TI - Activation of 1,1-difluoro-1-alkenes with a transition-metal complex: palladium(II)-catalyzed Friedel-Crafts-type cyclization of 4,4 (difluorohomoallyl)arenes. AB - Cationic palladium(II) ([Pd(MeCN)4](BF4)2) provides the first transition-metal catalyzed method for electrophilic activation of electron-deficient 1,1-difluoro 1-alkenes, which allows their Friedel-Crafts-type cyclization with an intramolecular aryl group via a Wacker-type process. By using BF3.OEt2, the cyclization was effected by a catalytic amount of the palladium without its reoxidation. PMID- 17914838 TI - Oxidative carbon-carbon bond formation via silyl bis-enol ethers: controlled cross-coupling for the synthesis of quaternary centers. AB - Unsymmetrical silyl bis-enol ethers have been developed as effective substrates for synthesizing quaternary centers from tetralone derivatives through oxidative carbon-carbon bond formation. The derived products are shown to be highly versatile intermediates that may be used to generate diverse structures such as cyclopentenones, 2H-pyrroles, and spirocyclic pyrrolidines. PMID- 17914839 TI - Selective defluorination approach to N-Cbz-3,3-difluoro-2 difluoromethylenepyrrolidine and its application to 3,3-difluoroproline dipeptide synthesis. AB - Mg-promoted defluorination of N-(p-methoxyphenyl)bis(trifluoromethyl)imine 1 gave perfluoroenamine 2, which was readily transformed to N-Cbz-2-trifluoromethyl-3,3 difluoropyrrolidine 10. Chemoselective defluorination from the trifluoromethyl group of 10 by LHMDS-promoted dehydrofluorination in THF provided 3,3-difluoro-2 difluoromethylenepyrrolidine 11. The product 11 was converted to 3,3 difluoroproline dipeptides 16 upon treatment with aminoesters. PMID- 17914840 TI - Novel permethylated beta-cyclodextrin derivatives appended with chromophores as efficient fluorescent sensors for the molecular recognition of bile salts. AB - Two novel permethylated beta-cyclodextrin (PM-beta-CD) derivatives, i.e., 6I-O-(1 naphtholxy)-2I,31-di-O-methylhexakis(2II-VII,3II-VII,6II-VII-tri-O-methyl)-beta cyclodextrin (1) and 6I-O-(8-hydroxyquinoline)-2I,31-di-O-methylhexakis(2II VII,3II-VII,6II-VII- tri-O-methyl)-beta-cyclodextrin (2), were synthesized in satisfactory yields, and their inclusion modes, complex-induced fluorescent behaviors, binding ability, and selectivity for bile salts of biological relevance (cholic acid sodium salt, CA; deoxycholic acid sodium salt, DCA; glycochoic acid sodium salt, GCA; taurocholic acid sodium salt, TCA) were investigated by the circular dichroism, 2D NMR, steady-state, and time-resolved fluorescent spectra. The results obtained from induced circular dichroism and ROESY spectra show that the chromophore groups of 1 and 2 reside in the central cavity of PM-beta-CD, and are expelled to the region of narrow torus rim upon complexation with bile guests, which presents the binding mode of cooperative inclusion. The transfer of the chromophore groups from the central cavity to the more hydrophobic torus rim leads to the remarkable increase of fluorescent intensities and longer fluorescent lifetimes of hosts 1 and 2 upon gradual addition of bile salts, which is importantly distinct from the molecular recognition of the chromophore-modified beta-CD species with bile salts. Interestingly, hosts 1 and 2 present much stronger binding ability for bile guests than PM-beta-CD. Differing from native beta-CD, all the PM-beta-CDs are more prone to include bile salts with longer tails, such as GCA and TCA. Their corresponding binding ability and molecular selectivity are closely discussed from the viewpoints of difference of cavity size/shape between beta-CD and PM beta-CD, effect of substituent groups, and structures of bile guests, respectively. PMID- 17914841 TI - Synthesis of archaeal bipolar lipid analogues: a way to versatile drug/gene delivery systems. AB - A synthetic route for the preparation of symmetrical and unsymmetrical archaeal tetraether-like analogues has been described. The syntheses are based upon the elaboration of hemimacrocyclic tetraether lipid cores from versatile building blocks followed by simultaneous or sequential introduction of polar head groups. Functionalizations of the tetraether lipids with neutral lactose or phosphatidylcholine polar heads and cationic glycine betaine moieties were envisaged both to increase membrane stability and to exhibit interactions with charged nucleic acids. Additionally, mannose and lactose triantennary clusters designed as multivalent ligands for selective interaction with lectin-type receptors were also efficiently synthesized for active cell/tissue targeting. PMID- 17914842 TI - Mechanism of SN2 disulfide bond cleavage by phosphorus nucleophiles. Implications for biochemical disulfide reducing agents. AB - The B3LYP variant of DFT has been used to study the mechanism of S-S bond scission in dimethyl disulfide by a phosphorus nucleophile, trimethylphospine (TMP). The reaction is highly endothermic in the gas phase and requires significant external stabilization of the charged products. DFT calculations (B3LYP) were performed with explicit (water molecules added) and implicit solvent corrections (COSMO model). The transition structures for this SN2 displacement reaction in a number of model systems have been located and fully characterized. The reaction barriers calculated with different approaches for different systems are quite close (around 11 kcal/mol). Remarkably, the calculations suggest that the reaction is almost barrierless with respect to the preorganized reaction complex and that most of the activation energy is required to rearrange the disulfide and TMP to its most effective orientation for the SMe group transfer way. Different reactivities of different phosphorus nucleophiles were suggested to be the result of steric effects, as manifested largely by varying amounts of hindrance to solvation of the initial product phosphonium ion. These data indicate that the gas-phase addition of a phosphine to the disulfide moiety will most likely form a phosphonium cation-thiolate anion salt, in the presence of four or more water molecules, that provide sufficient H-bonding stabilization to allow displacement of the thiolate anion, a normal uncomplicated SN2 transition state is to be expected. PMID- 17914843 TI - Oxime-based receptors for mono- and disaccharides. AB - Representatives of a new series of acyclic oxime-based receptors were prepared and their binding properties toward neutral sugar molecules studied. 1H NMR and fluorescence titrations revealed that receptors 2a and 2b, incorporating suitable positioned amine and oxime moieties, are able to form strong 1:1 complexes (Ka1 approximately 10(5) M-1) with dodecyl alpha- and beta-maltoside in chloroform solutions. Furthermore, the binding studies with beta-glucopyranoside indicated the formation of complexes with 1:1 and 1:2 receptor-monosaccharide binding stoichiometry (with overall binding constant beta2 approximately 10(5) M-2). Both hydrogen bonding and interactions of the sugar CH's with the phenyl rings of the receptor contribute to the stabilization of the receptor-sugar complexes. Molecular modeling calculations, synthesis, and binding studies are described. PMID- 17914844 TI - Stereochemical features making deoxycholic acid derived tropos biphenylphosphites efficient chiral ligands for rhodium: the asymmetric hydrogenation of dimethylitaconate as a case study. AB - Different deoxycholic acid derived biphenylphosphites, whose tropos nature was ascertained by NMR and CD measurements, were used in the rhodium-catalyzed asymmetric hydrogenation of dimethylitaconate achieving enantiomeric excesses up to 91%. The comparison of these results to those obtained using the corresponding atropoisomeric binaphthyl analogues, together with NMR and CD measurements on the rhodium complexes of some phosphites, allowed us to shed light on the nature of the active catalytic species and on the asymmetric induction process and hence to recognize the most appropriate stereochemical features to reach good levels of enantioselectivity. PMID- 17914845 TI - Molecular engineering of the internal charge transfer in thiophene-triphenylamine hybrid pi-conjugated systems. AB - Introduction of electroaccepting groups at the periphery of triphenylamine-based derivatives leads to an internal charge-transfer band. Syntheses and spectroscopic, electrochemical, and theoretical studies of various derivatives which differ by the strength and the number of electroacceptor groups are presented. These various results show that the ICT band and the acceptor/donor abilities of derivatives can be finely tuned. PMID- 17914847 TI - InCl3/I2-catalyzed cross-coupling of alkyl trimethylsilyl ethers and allylsilanes via an in situ derived combined Lewis acid of InCl3 and Me3SiI. AB - Direct Csp3-Csp3 coupling of various aliphatic trimethylsilyl ethers and allylsilanes is effectively catalyzed by InCl3 and I2. The transformation is thought to involve an in situ-derived combined Lewis acid of InCl3 and Me3SiI. The reaction can be used for the construction of quaternary-quaternary and quaternary-tertiary carbon-carbon bonds. This system enabled a highly chemoselective coupling to be conducted with a trimethylsilyl ether including an aryl halide moiety. Furthermore, couplings were possible using an alkynyltrimethylsilane and a trimethylsilyl ketene acetal. PMID- 17914846 TI - Synthesis and spectral properties of cholesterol- and FTY720-containing boron dipyrromethene dyes. AB - Two analogues (1, 2) of free cholesterol and one analogue (3) of the immunosuppressive sphingolipid FTY720 containing a boron dipyrromethene chromophore (BODIPY) were synthesized. The synthetic routes involved preparation of boron dipyrromethene moieties (5, 11), bearing a phenylethynyl group at different positions of the chromophore, and lipids (13, 20) bearing an azido group. The dye was tethered to the lipid via a 1,2,3-triazole in the linker by the click reaction. Analogues derived from 11 [in which an (E)-styrylethynyl moiety is bonded to C-5 of BODIPY] exhibited a marked red shift (approximately 70 80 nm) compared with those derived from 5 (in which a phenylethynyl moiety is bonded to C-8 of BODIPY). PMID- 17914848 TI - Structure formation in bis(terpyridine) derivative adlayers: molecule-substrate versus molecule-molecule interactions. AB - The influence of the substrate and the deposition conditions-vapor deposition versus deposition from solution-on the structures formed upon self-assembly of deposited bis(terpyridine) derivative (2,4'-BTP) monolayers on different hexagonal substrates, including highly oriented pyrolytic graphite (HOPG), Au(111), and (111)-oriented Ag thin films, was investigated by high-resolution scanning tunneling microscopy and by model calculations of the intermolecular energies and the lateral corrugation of the substrate-adsorbate interaction. Similar quasi-quadratic network structures with almost the same lattice constants obtained on all substrates are essentially identical to the optimum configuration expected from an optimization of the adlayer structure with C-H...N-type bridging bonds as a structure-determining factor, which underlines a key role of the intermolecular interactions in adlayer order. Slight distortions from the optimum values to form commensurate adlayer structures on the metal substrates and the preferential orientation of the adlayer with respect to the substrate are attributed to the substrate-adsorbate interactions, specifically, the lateral corrugation in the substrate-adsorbate interaction upon lateral displacement and rotation of the adsorbed BTP molecules. The fact that similar adlayer structures are obtained on HOPG under ultrahigh vacuum conditions (solid|gas interface) and on HOPG in trichlorobenzene (solid|liquid interface) indicates that the intermolecular interactions are not severely affected by the solvent. PMID- 17914849 TI - Adsorption and disjoining pressure isotherms of confined polymers using dissipative particle dynamics. AB - The adsorption and disjoining pressure isotherms of polymers confined by planar walls are obtained using Monte Carlo (MC) simulations in the Grand Canonical (GC) ensemble in combination with the mesoscopic technique known as dissipative particle dynamics (DPD). Two models of effective potentials for the confining surfaces are used: one with both an attractive and a repulsive term and one with a purely repulsive term. As for the polymer, seven-bead linear model of polyethylene glycol (PEG) dissolved in water is used. The results indicate remarkably good agreement between the trends shown by our adsorption isotherms and those obtained from experiments of PEG on oxide surfaces. Additionally, the disjoining pressure isotherm of water shows oscillations, while those of PEG display the same trend for both wall models. Moreover, it is found that the disjoining pressure isotherms are in qualitative agreement with those from experiments on confined linear polymers. PMID- 17914850 TI - Synthesis, characterization, and optical properties of well-defined N-doped, hollow silica/titania hybrid microspheres. AB - Well-defined nitrogen-doped, hollow SiO2/TiO2 hybrid spheres were successfully prepared through a two-step sol-gel synthesis combined calcination process using triethylamine as the nitrogen source. In this approach, polystyrene (PS)/silica microspheres were first synthesized. Subsequently, the amine-treated PS/SiO2/TiO2 hybrid spheres were obtained by sol-gel method. Finally, the elimination of the PS core, nitrogen-doping process, and crystallization of amorphous TiO2 were simultaneously conducted in the calcination process to acquire the final products. The as-prepared hybrid spheres were characterized by scanning electron microscopy, transmission electron microscopy, X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared (FTIR) spectroscopy. The results of XRD, FTIR, and XPS spectra indicated that nitrogen was really doped into the anatase TiO2 shell and confirmed that most nitrogen dopants might be present in the chemical environments of N-Ti-O and Ti-N-O. It was found that the absorption shoulder of nitrogen-doped hollow SiO2/TiO2 hybrid spheres vastly shifted to the visible region up to around 530 nm. The photoluminescence (PL) bands showed spectral lines at about 421, 472, and 529 nm, which were attributed to the self-trapped excitons, F and F+ centers. Moreover, the intensity of the PL spectra band of hollow SiO2/TiO2 hybrid spheres increased with as the amount of titanium tetrabutoxide (TBOT) precursor increased. However, the doping of nitrogen into hollow SiO2/TiO2 hybrid spheres led to the drastic quenching of photoluminescence because of the increase in the separation efficiency of the photoinduced electron and hole pairs. PMID- 17914851 TI - Fluorescence images of DNA-bound YOYO between coupled silver particles. AB - Oligonucleotide-bound silver particles were coupled through hybridization with target complementary oligonucleotides. YOYO molecules were intercalated into DNA duplexes bound between the coupled metal particles. Fluorescence images of YOYO molecules were monitored by scanning confocal microscopy. Relative to the free single YOYO, the emission brightness of the image was enhanced 80-fold after intercalating the fluorophores into the DNA duplexes between the coupled silver particles. Some images of the labeled metal particle dimers were observed to be dumbbell-shaped, suggesting that the stretching of intercalated YOYO molecules was restricted because of the orientation effect of fluorophores. The shortened lifetime of YOYO molecules between the coupled metal particles indicated that the fluorescence was enhanced via a near-field interaction mechanism between the fluorophore and the metal nanoparticle. PMID- 17914852 TI - Direct observation of Brownian dynamics of hard colloidal nanorods. AB - We synthesize monodisperse selenium (Se) colloidal rods, and the suspensions exhibit the smectic phase at a particle volume fraction (phi) of 0.28. Side-by side rod clustering occurs at phi > 0.04. Cluster-size distributions and persistence times are determined for various phi. In dense suspensions (phi > 0.1), individual rods reveal characteristic fundamental motions, e.g., reptation and synchronized rotation. Mean-square displacements of the rods suggest a cage trapping and escape. Estimated translational and rotational diffusion coefficients show a large difference from predictions by computer simulations. PMID- 17914853 TI - Importance of the Debye screening length on nanowire field effect transistor sensors. AB - Nanowire field effect transistors (NW-FETs) can serve as ultrasensitive detectors for label-free reagents. The NW-FET sensing mechanism assumes a controlled modification in the local channel electric field created by the binding of charged molecules to the nanowire surface. Careful control of the solution Debye length is critical for unambiguous selective detection of macromolecules. Here we show the appropriate conditions under which the selective binding of macromolecules is accurately sensed with NW-FET sensors. PMID- 17914854 TI - Fabrication of multiplex quasi-three-dimensional grids of one-dimensional nanostructures via stepwise colloidal lithography. AB - By using O2-plasma etched monolayers of hexagonally close-packed latex spheres as masks for metal vapor deposition, we successfully demonstrate a stepwise colloidal lithography to stepwise grow highly ordered multiplex quasi-three dimensional grids of metallic one-dimensional nanostructures, e.g., nanowires and nanorods. The success of the present approach is centered at manipulation of the incidence angle of metal vapor beams with respect to the normal direction of colloidal masks and particularly the azimuth angle phi of the projection of the vapor beam incidence on the masks with respect to the vector from one sphere to the nearest neighbors. Stepwise deposition of different metals by regularly varying phi allows consecutively stacking of 1D nanostructures into multiplex quasi-3D grids. This stepwise angle-resolved colloidal lithography should provide a significant nanochemical complement of conventional lithographic techniques, enabling us to easily fabricate sophisticated 3D nanostructures with defined vertical and lateral heterogeneity. PMID- 17914855 TI - Correlating dynamics and selectivity in adsorption of semiconductor nanocrystals onto a self-organized pattern. AB - Selective adsorption of semiconductor nanocrystals onto an organic self-organized pattern shows a time-dependent behavior. By studying the wetting behavior of delivered solvent (1-phenyloctane) on a lipid self-organized pattern and determining the adhesion energy between semiconductor nanocrystals and substrate, we obtain a correlation between dynamics and selectivity in adsorption of semiconductor nanocrystals onto the pattern by constructing a potential energy landscape. Two consecutive steps for selective adsorption of nanocrystals onto the self-organized pattern have been established: the first one is the molecule exchange of 1-phenyloctane and lipid molecules to form the adsorption sites for nanocrystals, and the second one is the adsorption of nanocrystals onto the adsorption sites due to the strong interaction between nanocrystals and substrate. PMID- 17914856 TI - Magnetic and electronic properties of CaCu3Cr4O12 and CaCu3Cr2Sb2O12 by first principles density functional calculation. AB - The electronic and magnetic properties of CaCu3Cr4O12 and CaCu3Cr2Sb2O12 are investigated by the use of the full-potential linearized augumented plane wave (FPLAPW) method. The calculated results indicate that CaCu3Cr4O12 is a ferrimagnetic and half-metallic compound, in good agreement with previous theoretical studies. CaCu3Cr2Sb2O12 is a ferrimagnetic semiconductor with a small gap of 0.136 eV. In both compounds, because Cr4+ 3d (d2) and Cr3+ 3d (d3) orbitals are less than half filled, the coupling between Cr-Cu is antiferromagnetic, whereas that between Cu-Cu and Cr-Cr is ferromagnetic. The total net spin moment is 5.0 and 3.0 microB for CaCu3Cr4O12 and CaCu3Cr2Sb2O12, respectively. In CaCu3Cr4O12, the 3d electrons of Cr4+ are delocalized, which strengthens the Cr-Cr ferromagnetic coupling. For CaCu3Cr2Sb2O12, the doping of nonmagnetic ion Sb5+ reduces the Cr-Cr ferromagnetic coupling, and the half filled Cr3+ t2g (t2g3) makes the chromium 3d electrons localized. In addition, the ordering arrangement of the octahedral chromium and antimony ions also prevents the delocalization of electrons. Hence, CaCu3Cr2Sb2O12 shows insulating behavior, in agreement with the experimental observation. PMID- 17914857 TI - Structural and kinetic study of reversible CO2 fixation by dicopper macrocyclic complexes. From intramolecular binding to self-assembly of molecular boxes. AB - A study of the reversible CO2 fixation by a series of macrocyclic dicopper complexes is described. The dicopper macrocyclic complexes [Cu2(OH)2(Me2p)](CF3SO3)2, 1(CF3SO3)2, and [Cu2(mu-OH)2(Me2m)](CF3SO3)2, 2(CF3SO3)2, (Scheme 1) containing terminally bound and bridging hydroxide ligands, respectively, promote reversible inter- and intramolecular CO2 fixation that results in the formation of the carbonate complexes [{Cu2(Me2p)}2(mu CO3)2](CF3SO3)4, 4(CF3SO3)4, and [Cu2(mu-CO3)(Me2m)](CF3SO3)2, 5(CF3SO3)2. Under a N2 atmosphere the complexes evolve CO2 and revert to the starting hydroxo complexes 1(CF3SO3)2 and 2(CF3SO3)2, a reaction the rate of which linearly depends on [H2O]. In the presence of water, attempts to crystallize 5(CF3SO3)2 afford [{Cu2(Me2m)(H2O)}2(mu-CO3)2](CF3SO3)4, 6(CF3SO3)4, which appears to rapidly convert to 5(CF3SO3)2 in acetonitrile solution. [Cu2(OH)2(H3m)]2+, 7, which contains a larger macrocyclic ligand, irreversibly reacts with atmospheric CO2 to generate cagelike [{Cu2(H3m)}2(mu-CO3)2](ClO4)4, 8(ClO4)4. However, addition of 1 equiv of HClO4 per Cu generates [Cu2(H3m)(CH3CN)4]4+ (3), and subsequent addition of Et3N under air reassembles 8. The carbonate complexes 4(CF3SO3)4, 5(CF3SO3)2, 6(CF3SO3)4, and 8(ClO4)4 have been characterized in the solid state by X-ray crystallography. This analysis reveals that 4(CF3SO3)4, 6(CF3SO3)4, and 8(ClO4)4 consist of self-assembled molecular boxes containing two macrocyclic dicopper complexes, bridged by CO32- ligands. The bridging mode of the carbonate ligand is anti-anti-mu-eta1:eta1 in 4(CF3SO3)4, anti-anti-mu eta2:eta1 in 6(CF3SO3)4 and anti-anti-mu-eta2:eta2 in 5(CF3SO3)2 and 8(ClO4)4. Magnetic susceptibility measurements on 4(CF3SO3)4, 6(CF3SO3)4, and 8(ClO4)4 indicate that the carbonate ligands mediate antiferromagnetic coupling between each pair of bridged CuII ions (J = -23.1, -108.3, and -163.4 cm-1, respectively, H = -JS1S2). Detailed kinetic analyses of the reaction between carbon dioxide and the macrocyclic complexes 1(CF3SO3)2 and 2(CF3SO3)2 suggest that it is actually hydrogen carbonate formed in aqueous solution on dissolving CO2 that is responsible for the observed formation of the different carbonate complexes controlled by the binding mode of the hydroxy ligands. This study shows that CO2 fixation can be used as an on/off switch for the reversible self-assembly of supramolecular structures based on macrocyclic dicopper complexes. PMID- 17914858 TI - Interaction of tertiary phosphines with lignin-type, alpha,beta-unsaturated aldehydes in water. AB - To learn more about the bleaching action of pulps by (hydroxymethyl)phosphines, lignin chromophores, such as the alpha,beta-unsaturated aromatic aldehydes, sinapaldehyde, coniferylaldehyde, and coumaraldehyde, were reacted with the tertiary phosphines R2R'P [R = R' = Me, Et, (CH2)3OH, iPr, cyclo-C6H11, (CH2)2CN; R = Me or Et, R' = Ph; R = Ph, R' = Me, m-NaSO3-C6H4] in water at room temperature under argon. In all cases, initial nucleophilic attack of the phosphine occurs at the activated C=C bond to form a zwitterionic monophosphonium species. With the phosphines PR3 [R = Me, Et, (CH2)3OH] and with R2R'P (R = Me or Et, R' = Ph), the zwitterion undergoes self-condensation to give a bisphosphonium zwitterion that can react with aqueous HCl to form the corresponding dichloride salts (as a mixture of R,R- and S,S-enantiomers); X-ray structures are presented for the bisphosphonium chlorides synthesized from the Et3P and Me3P reactions with sinapaldehyde. With the more bulky phosphines, iPr3P, MePPh2, (cyclo C6H11)3P, and Na[Ph2P(m-SO3-C6H4)], only an equilibrium of the monophosphonium zwitterion with the reactant aldehyde is observed. The weakly nucleophilic [NC(CH2)2]3P does not react with sinapaldehyde. An analysis of some exceptional 1H NMR data within the prochiral phosphorus centers of the bisphosphonium chlorides is also presented. PMID- 17914859 TI - Medium optimization for the production of recombinant nattokinase by Bacillus subtilis using response surface methodology. AB - Nattokinase is a potent fibrinolytic enzyme with the potential for fighting cardiovascular diseases. Most recently, a new Bacillus subtilis/Escherichia coli (B. subtilis/E. coli) shuttle vector has been developed to achieve stable production of recombinant nattokinase in B. subtilis (Chen; et al. 2007, 23, 808 813). With this developed B. subtilis strain, the design of an optimum but cost effective medium for high-level production of recombinant nattokinase was attempted by using response surface methodology. On the basis of the Plackett Burman design, three critical medium components were selected. Subsequently, the optimum combination of selected factors was investigated by the Box-Behnken design. As a result, it gave the predicted maximum production of recombinant nattokinase with 71 500 CU/mL for shake-flask cultures when the concentrations of soybean hydrolysate, potassium phosphate, and calcium chloride in medium were at 6.100, 0.415, and 0.015%, respectively. This was further verified by a duplicated experiment. Moreover, the production scheme based on the optimum medium was scaled up in a fermenter. The batch fermentation of 3 L was carried out by controlling the condition at 37 degrees C and dissolved oxygen reaching 20% of air saturation level while the fermentation pH was initially set at 8.5. Without the need for controlling the broth pH, recombinant nattokinase production with a yield of 77 400 CU/mL (corresponding to 560 mg/L) could be obtained in the culture broth within 24 h. In particular, the recombinant B. subtilis strain was found fully stable at the end of fermentation when grown on the optimum medium. Overall, it indicates the success of this experimental design approach in formulating a simple and cost-effective medium, which provides the developed strain with sufficient nutrient supplements for stable and high-level production of recombinant nattokinase in a fermenter. PMID- 17914860 TI - Engineering complex phenotypes in industrial strains. AB - The global demand is rising for greener manufacturing processes that are cost competitive and available in a timely manner. This has led to the development of a series of new tools and integrative platforms enabling rapid engineering of complex phenotypes in industrial microbes. By blending "old classical methods" of strain isolation with "newer approaches" of cell engineering, researchers are demonstrating the ability to stack multiple complex phenotypes in industrial hosts with some level of certainty. Newer tools for dissecting the genotype phenotype correlation include association analysis (Precision Engineering), multiSCale Analysis of Library Enrichment (SCALE) in competition experiments, whole-genome transcriptional analysis, and proteomics and metabolomics technology. These newer and older tools of metabolic engineering and synthetic biology when combined with recent whole cell engineering approaches like whole genome shuffling, global transciptome machinery engineering, and directed evolutionary engineering, provide a powerful platform for engineering complex phenotypes in industrial strains. This review attempts to highlight and compare these newer tools and approaches with traditional strain isolation procedures as it applies to genome engineering with examples taken from literature. PMID- 17914861 TI - Laser-induced heating of dextran-coated mesocapsules containing indocyanine green. AB - Indocyanine green (ICG) is a photosensitive reagent with clinically relevant diagnostic and therapeutic applications. Recently, ICG has been investigated for its utility as an exogenous chromophore during laser-induced heating. However, ICG's effectiveness remains hindered by its molecular instability, rapid circulation kinetics, and nonspecific systemic distribution. To overcome these limitations, we have encapsulated ICG within dextran-coated mesocapsules (MCs). Our objective in this study was to explore the ability of MCs to induce thermal damage in response to laser irradiation. To simulate tumorous tissue targeted with MCs, cylindrical phantoms were prepared consisting of gelatin, intralipid emulsion, and various concentrations of MCs. The phantoms were embedded within fresh chicken breast tissue representing surrounding normal tissue. The tissue models were irradiated at lambda = 808 nm for 10 min at constant power (P = 4.2 W). Five hypodermic thermocouples were used to record the temperature at various depths below the tissue surface and transverse distances from the laser beam central axis during irradiation. Temperature profiles were processed to remove the baseline temperature and influence of light absorption by the thermocouple and subsequently used to calculate a damage index based on the Arrhenius damage integral. Tissue models containing MCs experienced a maximum temperature change of 18.5 degrees C. Damage index calculations showed that the heat generation from MCs at these parameters is sufficient to induce thermal damage, while no damage was predicted in the absence of MCs. ICG maintains its heat-generating capabilities in response to NIR laser irradiation when encapsulated within MCs. Such encapsulation provides a potentially useful methodology for laser-induced therapeutic strategies. PMID- 17914862 TI - Novel orbital shake bioreactors for transient production of CHO derived IgGs. AB - Large-scale transient gene expression in mammalian cells is being developed for the rapid production of recombinant proteins for biochemical and preclinical studies. Here, the scalability of transient production of a recombinant human antibody in Chinese hamster ovary (CHO) cells was demonstrated in orbitally shaken disposable bioreactors at scales from 50 mL to 50 L. First, a small-scale multiparameter approach was developed to optimize the poly(ethylenimine)-mediated transfection in 50 mL shake tubes. This study confirmed the benefit, both in terms of extended cell culture viability and increased product yield, of mild hypothermic cultivation conditions for transient gene expression in CHO cells. Second, the scalability of the process was demonstrated in disposable shake bioreactors having nominal volumes of 5, 20, and 50 L with final antibody yields between 30 and 60 mg L(-1). Thus, the combination of transient gene expression with disposable shake bioreactors allows for rapid and cost-effective production of recombinant proteins in CHO cells. PMID- 17914863 TI - Separation of PEGylated alpha-lactalbumin from unreacted precursors and byproducts using ultrafiltration. AB - There is considerable clinical interest in the use of "second-generation" therapeutic proteins produced by conjugation of the native protein with various polymers including poly(ethylene glycol) (PEG). One of the challenges in the production of polymer-protein conjugates is the need to remove residual polymer, native (unreacted) protein, and any reaction byproducts from the final therapeutic formulation. The overall objective of this study was to evaluate the possibility of using ultrafiltration for the purification of a model PEGylated protein. Sieving data were obtained using PEGylated alpha-lactalbumin, the native protein, and the poly(ethylene glycol) over a range of pH, ionic strength, and filtrate flux using both neutral and charge-modified composite regenerated cellulose membranes. Purification of the PEGylated protein was achieved using a two-stage diafiltration process. The first stage used a neutral membrane to remove the unreacted protein and any small reaction byproducts while retaining the large PEGylated product. The second stage used a negatively charged membrane to remove the neutral poly(ethylene glycol) while retaining the PEGylated alpha lactalbumin as a result of strong electrostatic interactions. These results clearly demonstrate the potential of using membrane-based separations for the purification of second-generation therapeutic proteins. PMID- 17914865 TI - Bidirectional ion transfer between quadrupole arrays: MSn ion/ion reaction experiments on a quadrupole/time-of-flight tandem mass spectrometer. AB - Methods for bidirectional ion transmission between distinct quadrupole arrays were developed on a quadrupole/time-of-flight tandem mass spectrometer (QqTOF) containing three quadrupoles (ion guide Q0, mass filter Q1, and collision cell Q2) and a reflectron TOF analyzer, for the purpose of implementing multistage ion/ion reaction experiments. The transfer efficiency, defined as the percentage of ions detected after two transfer steps relative to the initial ion abundance, was found to be about 60% between Q2 and Q0 (with passage through the intermediate array (Q1)) and almost 100% between Q2 and Q1. Efficient ion transfer enabled new means for executing MSn experiments on an instrument of this type by operating Q1 in rf/dc mode for performing multiple steps of precursor/product ion isolation while passing ions through Q1 or trapping ions in Q1. In the latter case, the Q1 functioned as a linear ion trap. Either collision induced dissociation (CID) or ion/ion reactions can be conducted in between each stage of mass analysis. MS3 or MS4 experiments were developed to illustrate the charge increase of peptide ions via two steps of charge inversion ion/ion reactions, CID of electron-transfer dissociation (ETD) products and CID of a metal-peptide complex formed from ion/ion reactions. PMID- 17914864 TI - Analytical performance of a venturi-assisted array of micromachined ultrasonic electrosprays coupled to ion trap mass spectrometry for the analysis of peptides and proteins. AB - The analytical characterization of a novel ion source for mass spectrometry named array of micromachined ultrasonic electrosprays (AMUSE) is presented here. This is a fundamentally different type of ion generation device, consisting of three major components: (1) a piezoelectric transducer that creates ultrasonic waves at one of the resonant frequencies of the sample-filled device, (2) an array of pyramidally shaped nozzles micromachined on a silicon wafer, and (3) a spacer which prevents contact between the array and transducer ensuring the transfer of acoustic energy to the sample. A high-pressure gradient generated at the apexes of the nozzle pyramids forces the periodic ejection of multiple droplet streams from the device. With this device, the processes of droplet formation and droplet charging are separated; hence, the limitations of conventional electrospray-type ion sources, including the need for high charging potentials and the addition of organic solvent to decrease surface tension, can be avoided. In this work, a Venturi device is coupled with AMUSE in order to increase desolvation, droplet focusing, and signal stability. Results show that ionization of model peptides and small tuning molecules is possible with dc charging potentials of 100 Vdc or less. Ionization in rf-only mode (without dc biasing) was also possible. It was observed that, when combined with AMUSE, the Venturi device provides a 10-fold gain in signal-to-noise ratio for 90% aqueous sample solutions. Further reduction in the diameter of the orifices of the micromachined arrays led to an additional signal gain of at least 3 orders of magnitude, a 2-10-fold gain in the signal-to noise ratio and an improvement in signal stability from 47% to 8.5% RSD. The effectiveness of this device for the soft ionization of model proteins in aqueous media, such as cytochrome c, was also examined, yielding spectra with an average charge state of 8.8 when analyzed with a 100 Vdc charging potential. Ionization of model proteins was also possible in rf-only mode. PMID- 17914866 TI - Non-native heme-histidine ligation promotes microsecond time scale secondary structure formation in reduced horse heart cytochrome c. AB - Previous far-UV time-resolved optical rotatory dispersion (TRORD) studies of the sub-millisecond (burst) phase of secondary structure formation in horse and tuna cytochromes c after photoreduction in denaturant suggested that the non-native His18-Fe-His33 heme ligation dominant in the unfolded horse protein facilitated this fast folding better than did the His18-Fe-His26 coordination dominant in tuna [Chen, E., Goldbeck, R.A., and Kliger, D.S. (2003) J. Phys. Chem. A 107, 8149-8155; Chen, E., Goldbeck, R.A., and Kliger, D.S. (2004) J. Am. Chem. Soc. 126, 11175-11181]. Whether His18-Fe-His33 coordination actually facilitates fast secondary structure formation or just slows folding less than His18-Fe-His26 coordination is probed by examining the double histidine mutant H26QH33N of horse heart cytochrome c. The fast folding phase is absent in H26QH33N, indicating that His18-Fe-His33 misligation does promote fast secondary structure formation, as does His18-Fe-His26 to a lesser extent. His33 may be better able to facilitate folding because it is not as constrained by hydrogen bonding interactions in the denatured state as is His26. A greater flexibility, not only because of weakened or disrupted Van der Waals interactions in the presence of guanidine hydrochloride (GuHCl) but also because of its position relative to His18, may allow His33 to ligate to the heme group more easily than His26. These results are discussed along with the results of far-UV CD and Soret and visible region MCD measurements, which were performed to probe heme ligation in H26QH33N and to understand how GuHCl affects its folding stability and cooperativity. PMID- 17914867 TI - Strain and near attack conformers in enzymic thiamin catalysis: X-ray crystallographic snapshots of bacterial transketolase in covalent complex with donor ketoses xylulose 5-phosphate and fructose 6-phosphate, and in noncovalent complex with acceptor aldose ribose 5-phosphate. AB - Transketolase is a prominent thiamin diphosphate-dependent enzyme in sugar metabolism that catalyzes the reversible transfer of a 2-carbon dihydroxyethyl fragment between a donor ketose and an acceptor aldose. The X-ray structures of transketolase from E. coli in a covalent complex with donor ketoses d-xylulose 5 phosphate (X5P) and d-fructose 6-phosphate (F6P) at 1.47 A and 1.65 A resolution reveal significant strain in the tetrahedral cofactor-sugar adducts with a 25-30 degrees out-of-plane distortion of the C2-Calpha bond connecting the substrates' carbonyl with the C2 of the cofactor's thiazolium part. Both intermediates adopt very similar extended conformations in the active site with a perpendicular orientation of the scissile C2-C3 sugar bond relative to the thiazolium ring. The sugar-derived hydroxyl groups of the intermediates form conserved hydrogen bonds with one Asp side chain, with a cluster of His residues and with the N4' of the aminopyrimidine ring of the cofactor. The phosphate moiety is held in place by electrostatic and hydrogen-bonding interactions with Arg, His, and Ser side chains. With the exception of the thiazolium part of the cofactor, no structural changes are observable during intermediate formation indicating that the active site is poised for catalysis. DFT calculations on both X5P-thiamin and X5P thiazolium models demonstrate that an out-of-plane distortion of the C2-Calpha bond is energetically more favorable than a coplanar bond. The X-ray structure with the acceptor aldose d-ribose 5-phosphate (R5P) noncovalently bound in the active site suggests that the sugar is present in multiple forms: in a strained ring-closed beta-d-furanose form in C2-exo conformation as well as in an extended acyclic aldehyde form, with the reactive C1 aldo function held close to Calpha of the presumably planar carbanion/enamine intermediate. The latter form of R5P may be viewed as a near attack conformation. The R5P binding site overlaps with those of the leaving group moieties of the covalent donor-cofactor adducts, demonstrating that R5P directly competes with the donor-derived products glyceraldehyde 3-phosphate and erythrose 4-phosphate, which are substrates of the reverse reaction, for the same docking site at the active site and reaction with the DHEThDP enamine. PMID- 17914868 TI - Thermodynamic dissection of the Ezrin FERM/CERMAD interface. AB - ERM (Ezrin-Radixin-Moesin) proteins are key cross-linkers of the plasma membrane and the actin cytoskeleton. They are regulated by the intramolecular association of the N-terminal FERM (band-four point one, Ezrin, Radixin, Moesin) and C terminal CERMAD (ERM association domain) domains (N/C interaction), which masks the binding surfaces of the domains for other molecules. The N/C interface is characterized by the highly distributed binding of CERMAD through a beta-strand and four alpha-helices to a globular FERM. Though it is a target for multiple regulatory signals, little is known about the dynamics/thermodynamics governing this interface. Recent implications of Ezrin in cancer metastasis have increased the necessity to understand this regulatory switch. In this study, we report residue-specific stabilities of Ezrin CERMAD at the Ezrin N/C interface obtained using hydrogen-deuterium exchange NMR. These stabilities vary across secondary structural elements and identify F583 and L586 as key anchor residues for the most stable element, alphaD. Macroscopic N/C binding energetics, obtained using isothermal titration calorimetry (ITC) reveals a high affinity (Kd =176 nM) enthalpy-driven binding (DeltaH = -26 kcal/mol, TDeltaS = -17 kcal/mol) at 25 degrees C at pH 7 in MES and phosphate buffers. A 10-fold increase in affinity was observed for measurements in acetate buffer, suggesting that an acetate-like molecule might promote the repressed form of the complex, possibly through interaction with the F2 subdomain of FERM, which resembles the acyl-CoA binding protein. In summary, our results have illustrated the dynamic nature of this regulatory interface and provide a foundation for investigating the role of regulatory signals on the stability of this interface. PMID- 17914870 TI - Occurrence of orally administered mulberry 1-deoxynojirimycin in rat plasma. AB - 1-deoxynojirimycin (DNJ), a potent glucosidase inhibitor, is a characteristic constituent of the mulberry leaf. Dietary mulberry DNJ may be beneficial for the suppression of abnormally high blood glucose levels, thereby preventing diabetes mellitus. Although there is considerable interest in the effects of mulberry DNJ, the intestinal absorption and pharmacokinetic profile of orally administered mulberry DNJ have never been characterized. In this study, we developed a method for determining the level of plasma DNJ by hydrophilic interaction chromatography coupled to a mass spectrometric detector (HILIC-MS) to investigate the absorption and metabolism of orally administered mulberry DNJ in rats. DNJ was separated from plasma extract on a TSK gel Amide-80 column, a representative column for HILIC. At postcolumn, DNJ was concurrently detected and identified by MS. The plasma DNJ concentration in fasted rats was below the detection limit [<1 microg (6 nmol)/mL]; however, the concentration reached a maximum [15 microg (92 nmol)/mL] 30 min after the administration of mulberry DNJ (110 mg/kg of body weight), and the DNJ concentration decreased rapidly thereafter. When the rats received different amounts of mulberry DNJ (1.1, 11, and 110 mg/kg of body weight), dose-dependent incorporation of DNJ into the plasma was confirmed. We did not detect any DNJ metabolites in the plasma. These findings indicate that orally administered mulberry DNJ is absorbed as an intact form from the alimentary tract and then is quickly excreted from the body. The developed HILIC MS method could be applied in determining levels of DNJ in urine and tissues, and therefore, the method would be a powerful tool for studying the metabolic fate of mulberry DNJ as well as its bioavailability. PMID- 17914869 TI - Cytotoxicity of polyspermine-ribonuclease A and polyspermine-dimeric ribonuclease A. AB - Polyspermine-ribonuclease A (PS-RNase A) and polyspermine-dimeric ribonuclease A (PS-dimeric RNase A) were prepared by cross-linking ribonuclease A or its covalently linked dimer to polyspermine (PS) using dimethyl suberimidate. The two RNase A derivatives were tested for a possible antitumor action. The in vitro and in vivo cytotoxic activity of PS-RNase A, although strong, is not higher than that known for free polyspermine. PS-dimeric RNase A, which was characterized by mass spectroscopy, titration of free amine groups, and enzymatic assays, proved instead to be a definitely more efficient antitumor agent, both in vitro and in vivo. This result could tentatively be explained in view of the importance of positive charges for ribonuclease activity, considering the higher basicity of PS dimeric RNase A compared to that of PS-(monomeric)RNase A. It must be also taken into account that the dimeric RNase A moiety of PS-dimeric RNase A could evade the cytoplasmic ribonuclease inhibitor, which instead could trap the monomeric RNase A moiety of the other derivative. The two RNase A derivatives degrade poly(A).poly(U) under conditions where native RNase A is inactive. The results of this work demonstrate once again the importance of positive charges for the functions of mammalian pancreatic type ribonucleases in general, in particular for RNase A derivatives, and the potential therapeutic use of the ribonuclease A derivatives. PMID- 17914871 TI - Degradation of cyanidin 3-rutinoside in the presence of (-)-epicatechin and litchi pericarp polyphenol oxidase. AB - The degradation mechanism of cyanidin 3-rutinoside in the presence of (-) epicatechin and litchi pericarp polyphenol oxidase (PPO) was investigated using several model systems. The enzymically generated (-)-epicatechin o-quinone could induce cyanidin 3-rutinoside degradation. The results obtained in this study allowed us to propose a pathway for cyanidin 3-rutinoside degradation in the presence of (-)-epicatechin and litchi pericarp PPO. First, enzymatic oxidation of (-)-epicatechin produced the corresponding o-quinone, and then cyanidin 3 rutinoside and (-)-epicatechin competed for (-)-epicatechin o-quinone, resulting in degradation of cyanidin 3-rutinoside and regeneration of (-)-epicatechin. Moreover, the results of kinetic studies indicated this competition was influenced by both (-)-epicatechin concentration and cyanidin 3-rutinoside concentration in the model system. PMID- 17914872 TI - Chemical composition of the volatile extract and antioxidant activities of the volatile and nonvolatile extracts of Egyptian corn silk (Zea mays L.). AB - A total of 36 compounds, which comprised 99.4% of the extract, were identified by gas chromatography and mass spectrometry (GC-MS) in the volatile dichloromethane extract obtained from Egyptian corn silk. The main constituents of the volatile extract were cis-alpha-terpineol (24.22%), 6,11-oxidoacor-4-ene (18.06%), citronellol (16.18%), trans-pinocamphone (5.86%), eugenol (4.37%), neo-iso-3 thujanol (2.59%), and cis-sabinene hydrate (2.28%). Dried Egyptian corn silk was also directly extracted with petroleum ether, ethanol, and water. All extracts from solvent extraction and the volatile extract described above exhibited clear antioxidant activities at levels of 50-400 microg/mL in the 2,2-diphenyl-1 picrylhydrazyl (DPPH)/linoleic acid assay. The ethanol extract inhibited DPPH activity by 84% at a level of 400 microg/mL. All samples tested via the beta carotene bleaching assay also exhibited satisfactory antioxidant activity with clear dose responses. This study indicates that corn silk could be used to produce novel natural antioxidants as well as a flavoring agent in various food products. PMID- 17914873 TI - Significant lability of guaiacylglycerol beta-phenacyl ether under alkaline conditions. AB - It was observed that the beta-O-4 bond cleavage of a dimeric phenolic lignin model compound with an alpha-carbonyl group at the B-ring, 2-(2-ethoxy-4 formylphenoxy)-1-(4-hydroxy-3-methoxyphenyl)propane-1,3-diol (I), is extremely fast in a mild anaerobic alkaline treatment (0.45 mol/L NaOH, 95 degrees C, 0.8 MPa of N2). This phenomenon significantly contrasts with the case of a common dimeric phenolic lignin model compound without any specific functional group, 1 (4-hydroxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)propane-1,3-diol (II). The most plausible mechanism is the migration of the B-ring from the beta- to the alpha position following the SNAr mechanism. Because this migration affords the alkaline labile phenolic alpha-O-4-type compound (XI), the formation of the quinone methide as well as the cleavage of the originally alkaline very stable alkyl-aryl ether bond is promoted. This promotion of the quinone methide formation explains why a relatively large amount of 4-hydroxy-3 methoxybenzaldehyde (IV) is produced from I in an oxygen-alkali treatment. PMID- 17914874 TI - Simple multiresidue method for monitoring of trimethoprim and sulfonamide residues in buffalo meat by high-performance liquid chromatography. AB - A simple, specific, and rapid analytical method for the determination of trimethoprim (TMP) and three sulfonamide (SA) antimicrobial drug residues in buffalo meat is developed and validated. This method is based on a solid-phase extraction technique followed by high-performance liquid chromatography (HPLC) photodiode array (PDA) detection. Target compounds were extracted from the meat by acetonitrile and water, cleaned up on a Bond Elute C 18 cartridge column, and separated on a RP-C 18 column during HPLC analysis. Acetonitrile along with water appears to be an excellent extractant as recovery of the analytes at maximum residues levels (MRLs) in spiked sample was in the range of 75-108%, with coefficient of variations (CVs) ranging between 1.34 and 22%. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.031 and 0.062 microg/g, respectively, for all of the compounds. Intra- and interday assay precisions of the method at 0.125 microg/g concentrations for any drug ranged between 3 and 4%. The linearities of the TMP, sulfadimidine (SDM), sulfadoxine (SDO), and sulfamethoxazole (SMX) were 0.9989, 0.9999, 0.9998, and 0.9997, respectively. For robustness, the analytical method was applied to 122 buffalo meat samples obtained from export meat processing plants. PMID- 17914875 TI - Antioxidant activity, polyphenol content, and related compounds in different fruit juices and homogenates prepared from 29 different pomegranate accessions. AB - Pomegranate juice is well known for its health beneficial compounds, which can be attributed to its high level of antioxidant activity and total polyphenol content. Our objective was to study the relationships between antioxidant activity, total polyphenol content, total anthocyanins content, and the levels of four major hydrolyzable tannins in four different juices/homogenates prepared from different sections of the fruit. To this end, 29 different accessions were tested. The results showed that the antioxidant activity in aril juice correlated significantly to the total polyphenol and anthocyanin contents. However, the homogenates prepared from the whole fruit exhibited an approximately 20-fold higher antioxidant activity than the level found in the aril juice. Unlike the arils, the antioxidant level in the homogenates correlated significantly to the content of the four hydrolyzable tannins in which punicalagin is predominant, while no correlation was found to the level of anthocyanins. PMID- 17914876 TI - Designed polar cosolvent-modified supercritical CO2 removing caffeine from and retaining catechins in green tea powder using response surface methodology. AB - This study examines cosolvent-modified supercritical carbon dioxide (SC-CO2) to remove caffeine from and to retain catechins in green tea powder. The response surface method was adopted to determine the optimal operation conditions in terms of the extraction efficiencies and concentration factors of caffeine and catechins during the extractions. When SC-CO2 was used at 333 K and 300 bar, 91.5% of the caffeine was removed and 80.8% of catechins were retained in the tea: 3600 g of carbon dioxide was used in the extraction of 4 g of tea soaked with 1 g of water. Under the same extraction conditions, 10 g of water was added to <800 g of carbon dioxide in an extraction that completely removed caffeine (that is, the caffeine extraction efficiency was 100%). The optimal result as predicted by three-factor response surface methodology and supported by experimental data was that in 1.5 h of extraction, 640 g of carbon dioxide at 323 K and 275 bar with the addition of 6 g of water extracted 71.9% of the caffeine while leaving 67.8% of the catechins in 8 g of tea. Experimental data indicated that supercritical carbon dioxide decaffeination increased the concentrations of caffeine in the SC-CO2 extracts at 353 K. PMID- 17914877 TI - Factors affecting the retention of rennet in cheese curd. AB - The coagulant retained in cheese curd is a major contributor to proteolysis during ripening. The objective of this study was to quantify the effects of several milk-related factors and parameters during cheese manufacture on the retention of coagulant in cheese curd. The amount of coagulant retained in curd was determined by its activity on a synthetic heptapeptide (Pro-Thr-Glu-Phe-[NO2 Phe]-Arg-Leu) using reversed-phase HPLC. The retention of chymosin in cheese curd increased significantly when the pH of milk was reduced at rennet addition below pH 6.1, the pH at whey drainage below pH 5.7, or the average casein micelle size in milk and when the ionic strength of milk was increased. The casein content of milk and the quantity of chymosin added to milk had no significant effect on the retention of chymosin in curd; the quantity of coagulant bound per gram of casein remained unchanged. PMID- 17914878 TI - Polyphenols distribution in juices from Citrus allotetraploid somatic hybrids and their sexual hybrids. AB - The polyphenol profile of an interspecific allotetraploid somatic hybrid, achieved by protoplast fusion, the 'Valencia' sweet orange ( Citrus sinensis L. Osbeck) + 'Femminello' lemon ( Citrus limon L. Burm), and three sexual hybrids obtained by backcrosses between Femminello lemon and the allotetraploid somatic hybrid ('Valencia' + 'Femminello') was studied by liquid chromatography ultraviolet-diode array detector-mass spectrometry (LC-UV-DAD-MS). The aim of the work was to evaluate whether superior traits and improved performance can be observed in these new genotypes. Ten flavonoids (TF), comprising seven flavanones and three flavones, and four hydroxycinnamic acids (HCA) have been characterized and quantified in all hybrids and compared with those of the respective parents. The 'Valencia' + 'Femminello' somatic hybrid shows an intermediate polyphenol composition with respect to those of the parents, with a slight prevalence of lemon influence. The three sexual hybrids show, instead, different and more complex chromatographic profiles. PMID- 17914879 TI - Peptides derived from dolicholin, a phaseolin-like protein in country beans (Dolichos lablab), potently stimulate cholecystokinin secretion from enteroendocrine STC-1 cells. AB - Peptides derived from soybean beta-conglycinin and pork protein stimulate cholecystokinin (CCK) secretion from the enteroendocrine cells (EECs) and suppress food intake. Here we examined CCK-releasing activities from the enteroendocrine cell line STC-1, in peptides derived from underutilized legumes, and found much higher activity in the peptic hydrolysate of Country beans (CBP) compared to that from other legume-derived peptides including beta-conglycinin peptone. Active components in CBP were separated into acetonitrile-soluble fractions, but the activities were abolished after pronase treatment. To identify the Country bean protein containing the active peptides, Country bean protein extracts in an alkaline solution (CBE) were fractionated based on isoelectric point or molecular weight. Peptones prepared from CBE fractions containing a 51 kDa major protein stimulated CCK release, but other fractions did not. N-Terminal sequence analysis indicated that the 51 kDa protein is a phaseolin-like globular protein, and we designated this protein dolicholin. These results indicate that Country bean-derived peptides are very potent legume peptides in stimulating CCK secretion from EECs and that the stimulant peptides originate from dolicholin, a newly identified phaseolin-like globular protein in Country beans. PMID- 17914880 TI - Inter- and intraspecific variations of cadmium accumulation of 13 leafy vegetable species in a greenhouse experiment. AB - Leafy vegetables are among the crop species that are most vulnerable to heavy metal pollution. This study investigated inter- and intraspecific variations of cadmium accumulation in 13 species with a total of 39 cultivars of leafy vegetables under two levels of soil Cd stress (1.5 and 7.7 mg kg (-1)). Intraspecific variations of shoot biomass and Cd concentration of the tested leafy vegetables were significantly larger than interspecific variations under both Cd treatments and were also more significantly correlated between two Cd stress levels when grouped by cultivar than grouped by species. These results indicate that cultivar is a more reliable taxa level for screening pollutant-safe leafy vegetables than species. Any screening for pollutant hypoaccumulator species, or other similar species-based concepts, without considering intraspecific variation should be avoided. PMID- 17914881 TI - Pavietin, a coumarin from Aesculus pavia with antifungal activity. AB - A new prenylated coumarin, S-6-[2-(hydroxymethyl)butoxy]-7-hydroxy-4-methyl-2 H chromen-2-one ( 1), named pavietin, has been isolated from the leaves of an Aesculus pavia genotype along with three known flavonol glycosides, quercetin 3- O-alpha-rhamnoside (quercitrin, 2), quercetin 3- O-alpha-arabinoside ( 3), and isorhamnetin 3- O-alpha-arabinoside (distichin, 4). The chemical structure of compound 1 was determined by chemical and spectroscopic methods, inclusive of UV, MS, and 1D and 2D NMR experiments. It showed appreciable antimicrobial properties against several pathogens, displaying a significant antifungal activity toward one of the main fungal parasites of Aesculus species, Guignardia aesculi. The same biological tests performed with a mixture of flavonoids 2- 4 resulted in weak or no activity. Compound 1 was undetectable in Aesculus hippocastanum, a closely related species lacking resistance to fungal pathogens. The possible role of 1 in plant resistance is discussed. PMID- 17914882 TI - Absolute configuration of 8-Amino-3,8-dideoxyoct-2-ulosonic acid, the chemical hallmark of lipopolysaccharides of the genus Shewanella. AB - The novel monosaccharide 8-amino-3,8-dideoxyoct-2-ulosonic acid (Kdo8N) was isolated by methanolysis from the lipooligosaccharide of the marine Gram-negative bacterium Shewanella pacifica. After HPLC purification, the absolute configuration was determined by the Mosher ester method and proven to be 4 R,5 R,6 R,7 R. This established the d- manno- configuration of the monosaccharide. PMID- 17914885 TI - Chemical and enzymatic oxidation of furosemide: formation of pyridinium salts. AB - Furosemide (Lasix) is frequently used in the treatment of cardiovascular and renal disease. Only one metabolite, furosemide glucuronide, has ever been identified. Oxidation of furosemide by cytochrome P450 has been demonstrated, but the metabolite(s) has never been identified. The oxidation of furosemide by dimethyldioxirane in acetone and by liver microsomal incubations was explored in this study. The first observable product from dimethyldioxirane oxidation was a ring-expanded enone resulting from an intramolecular condensation of the aldehyde group of the enonal, the secondary amine, and the carboxylic acid in a Mannich like reaction. Keto-enol tautomerization and opening of the lactone gave a stable pyridinium salt. The pyridinium salt was also observed in the microsomal incubations of furosemide. The presence of an internal nucleophile in furosemide may have a significant effect on the toxicology and possibly the pharmacology of this furan. PMID- 17914884 TI - Synthesis of oligonucleotides containing the N2-deoxyguanosine adduct of the dietary carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline. AB - 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) is a highly mutagenic heterocyclic amine formed in all cooked meats. IQ has been found to be a potent inducer of frameshift mutations in bacteria and carcinogenic in laboratory animals. Upon metabolic activation, IQ forms covalent adducts at the C8- and N2-positions of deoxyguanosine with a relative ratio of up to approximately 4:1. We have previously incorporated the major dGuo-C8-IQ adduct into oligonucleotides through the corresponding phosphoramidite reagent. We report here the sequence-specific synthesis of oligonucleotides containing the minor dGuo-N2-IQ adduct. Thermal melting analysis revealed that the dGuo-N2-IQ adduct significantly destabilizes duplex DNA. PMID- 17914886 TI - Fluorous and traceless synthesis of substituted indole alkaloids. AB - The fluorous traceless synthesis of substituted indole alkaloids is carried out first by attaching the 3-(perfluorooctyl)propanol with Boc protected L tryptophan. The reaction of perfluoroalkyl (Rfh)-tagged tryptophan esters with various aldehydes undergoes Pictet-Spengler reaction to give cis and trans stereoisomers of tetrahydro-beta-carbolines. The nucleophilic addition of the piperidine nitrogen across various isocyanates is followed by the cyclization of ureas and simultaneous rupture of the fluorous tag to afford the hydantoin ring fused tetrahydro-beta-carbolines. All the fluorous-tag compounds are purified by solid-phase extraction (SPE) through Fluoro Flash cartridges. PMID- 17914883 TI - Measurement of the incorporation and repair of exogenous 5-hydroxymethyl-2' deoxyuridine in human cells in culture using gas chromatography-negative chemical ionization-mass spectrometry. AB - The DNA of all organisms is constantly damaged by oxidation. Among the array of damage products is 5-hydroxymethyluracil, derived from oxidation of the thymine methyl group. Previous studies have established that HmU can be a sensitive and valuable marker of DNA damage. More recently, the corresponding deoxynucleoside, 5-hydroxymethyl-2'-deoxyuridine (HmdU), has proven to be valuable for the introduction of controlled amounts of a single type of damage lesion into the DNA of replicating cells, which is subsequently repaired by the base excision repair pathway. Complicating the study of HmU formation and repair, however, is the known chemical reactivity of the hydroxymethyl group of HmU under conditions used to hydrolyze DNA. In the work reported here, this chemical property has been exploited by creating conditions that convert HmU to the corresponding methoxymethyluracil (MmU) derivative that can be further derivatized to the 3,5 bis-(trifluoromethyl)benzyl analogue. This derivatized compound can be detected by gas chromatography-negative chemical ionization-mass spectrometry (GC-NCI-MS) with good sensitivity. Using isotopically enriched exogenous HmdU and human osteosarcoma cells (U2OS) in culture, we demonstrate that this method allows for the measurement of HmU in DNA formed from the incorporation of exogenous HmdU. We further demonstrate that the addition of isotopically enriched uridine to the culture medium allows for the simultaneous measurement of DNA replication and repair kinetics. This sensitive and facile method should prove valuable for studies on DNA oxidation damage and repair in living cells. PMID- 17914888 TI - Design of organocatalysts for asymmetric direct syn-aldol reactions. AB - Two new organocatalysts 3a and 3b, derived from L-leucine and (S)-beta-amino alcohols that were prepared from L-valine, were designed and afforded the direct syn-aldol reactions of a wide scope of aldehydes with various ketones with an excellent diastereomeric ratio of up to >20/1 and enantioselectivities of up to 99% ee. PMID- 17914887 TI - Simulations of ionic liquids, solutions, and surfaces. AB - We have been using atomistic simulation for the last 10 years to study properties of imidazolium-based ionic liquids. Studies of dissolved molecules show the importance of electrostatic interactions in both aromatic and hydrogen-bonding solutes. However, the local structure strongly depends upon ion-ion and solute solvent interactions. We find interesting local alignments of cations at the gas liquid and solid-liquid interfaces, which give a potential drop through the surface. If the solid interface is charged, this charge is strongly screened over distances of a few nanometres and this screening decays on a fast time scale. We have studied the sensitivity of the liquid structure to force-field parameters and show that results from ab initio simulations can be used in the development of force fields. PMID- 17914889 TI - In situ solution-phase Raman spectroscopy under forced convection. AB - In situ Raman spectra of solution-phase electrogenerated species have been recorded in a channel-type electrochemical cell incorporating a flat optically transparent window placed parallel to the channel plane that contains the embedded working electrode. A microscope objective with its main axis (Z) aligned normal to the direction of flow was used to focus the excitation laser beam (lambda exc = 532 nm) in the solution and also to collect the Raman scattered light from species present therein. Judicious adjustment of the cell position along Z allowed the depth of focus to overlap the diffusion boundary layer to achieve maximum detection sensitivity. Measurements were performed using a Au working electrode in iron hexacyanoferrate(II), [Fe(CN)6]4-, and nitrite, NO2-, containing aqueous solutions as a function of the applied potential, E. Linear correlations were found between both the gain and the loss of the integrated Raman intensity, IR, of bands, attributed to [Fe(CN)6]3- and [Fe(CN)6]4-, respectively, recorded downstream from the edge of the working electrode, and the current measured at the Au electrode as a function of E. The same overall trend was found for the gain in the IR of the NO3- band in the nitrite solution. Also included in this work is a ray trace analysis of the optical system. PMID- 17914891 TI - Miniaturization and geometry optimization of a polymer-based rectilinear ion trap. AB - The fabrication, operation, and characterization of a polymer-based rectilinear ion trap mass analyzer is discussed. A novel, fast prototyping technique, stereolithography (SLA)-based fabrication, traditionally reserved for end use production parts and to fabricate master molds for rubber products, is applied here as a tool to create precise, arbitrary geometries. Taking full advantage of the SLA methodology, an open corner, polymer-based ion trap has been fabricated and tested. The use of a custom resin, Nanoform 15120 (DSM Somos, New Castle, DE), has resulted in a polymer with high heat deflection temperature and greater structural stability at higher temperatures and lower capacitance. The mass analyzer was mounted in a polymer holder and tested in a custom vacuum system using modified LCQ Duo (Thermo Fisher Corp.) electronics. The resolution, mass/charge range, and MS/MS capabilities were examined using electrospray ionization as well as atmospheric pressure chemical ionization. In the course of this study, three traps of different sizes were fabricated, beginning with a "full size" device measuring 10 x 8 x 50 mm. The next two traps were scaled down by linear factors of a half and a third. SLA is shown to allow fabrication of light, small rectilinear ion traps, which are less expensive and have the same performance as traditional machined devices of the same size. In addition, smaller traps can be built just as easily, and they show unit mass resolution to mass 300, tandem mass spectrometry capabilities, and low power consumption. PMID- 17914890 TI - Laser-driven acoustic desorption of organic molecules from back-irradiated solid foils. AB - Laser-induced acoustic desorption (LIAD) from thin metal foils is a promising technique for gentle and efficient volatilization of intact organic molecules from surfaces of solid substrates. Using the single-photon ionization method combined with time-of-flight mass spectrometry, we have examined the neutral component of the desorbed flux in LIAD and compared it to that from direct laser desorption. These basic studies of LIAD, conducted for molecules of various organic dyes (rhodamine B, fluorescein, anthracene, coumarin, BBQ), have demonstrated detection of intact parent molecules of the analyte even at its surface concentrations corresponding to a submonolayer coating. In some cases (rhodamine B, fluorescein, BBQ), the parent molecular ion peak was accompanied by a few fragmentation peaks of comparable intensity, whereas for others, only peaks corresponding to intact parent molecules were detected. At all measured desorbing laser intensities (from 100 to 500 MW/cm2), the total amount of desorbed parent molecules depended exponentially on the laser intensity. Translational velocities of the desorbed intact molecules, determined for the first time in this work, were of the order of hundreds of meters per second, less than what has been observed in our experiments for direct laser desorption, but substantially greater than the possible perpendicular velocity of the substrate foil surface due to laser-generated acoustic waves. Moreover, these velocities did not depend on the desorbing laser intensity, which implies the presence of a more sophisticated mechanism of energy transfer than direct mechanical or thermal coupling between the laser pulse and the adsorbed molecules. Also, the total flux of desorbed intact molecules as a function of the total number of desorbing laser pulses, striking the same point on the target, decayed following a power law rather than an exponential function, as would have been predicted by the shake off model. To summarize, the results of our experiments indicate that the LIAD phenomenon cannot be described in terms of simple mechanical shake-off or direct laser desorption. Rather, they suggest that multistep energy-transfer processes are involved. Two possible (and not mutually exclusive) qualitative mechanisms of LIAD that are based on formation of nonequilibrium energy states in the adsorbate substrate system are proposed and discussed. PMID- 17914892 TI - Elemental analysis of organic species with electron ionization high-resolution mass spectrometry. AB - We present a new elemental analysis (EA) technique for organic species (CHNO) that allows fast on-line analysis (10 s) and reduces the required sample size to approximately 1 ng, approximately 6 orders of magnitude less than standard techniques. The composition of the analyzed samples is approximated by the average elemental composition of the ions from high-resolution electron ionization (EI) mass spectra. EA of organic species can be performed on organic/inorganic mixtures. Elemental ratios for the total organic mass, such as oxygen/carbon (O/C), hydrogen/carbon (H/C), and nitrogen/carbon (N/C), in addition to the organic mass to organic carbon ratio (OM/OC), can be determined. As deviations between the molecular and the ionic composition can appear due to chemical influences on the ion fragmentation processes, the method was evaluated and calibrated using spectra from 20 compounds from the NIST database and from 35 laboratory standards sampled with the high-resolution time-of-flight aerosol mass spectrometer (HR-ToF-AMS). The analysis of AMS (NIST) spectra indicates that quantification of O/C is possible with an error (average absolute value of the relative error) of 30% (17%) for individual species. Precision is much better than accuracy at +/-5% in the absence of air for AMS data. AMS OM/OC has an average error of 5%. Additional calibration is recommended for types of species very different from those analyzed here. EA was applied to organic mixtures and ambient aerosols (sampled at 20 s from aircraft). The technique is also applicable to other EI-HRMS measurements such as direct injection MS. PMID- 17914893 TI - Updated meta-analytical approach to the efficacy of antihypertensive drugs in reducing blood pressure. AB - BACKGROUND AND OBJECTIVE: Despite advances in the treatment of hypertension, control rates continue to be suboptimal in both Europe and the US. Strategies that improve hypertension control are therefore urgently needed. This study aimed to assess the relative efficacies of various antihypertensive drugs commonly used in France in reducing systolic and diastolic blood pressure (SBP and DBP) by using a meta-analytical approach. This update of a previously published meta analytical approach extends the number of drugs evaluated from 13 to 19. METHODS: A total of 80 randomised, controlled trials published between 1973 and 2007 involving 10 818 patients were selected for inclusion in the meta-analytical approach. Data were examined for 19 drugs, and 16 drugs were included in the analysis: hydrochlorothiazide, indapamide sustained-release (SR), atenolol, amlodipine, lercanidipine, manidipine, enalapril, ramipril, trandolapril, candesartan cilexetil, irbesartan, losartan, olmesartan medoxomil, telmisartan, valsartan and aliskiren. Weighted average reductions in SBP and DBP over a period of 8-12 weeks were calculated for each drug from information on both the mean and the variability in BP reduction. No trials evaluating furosemide, spironolactone or cicletanine satisfied the inclusion criteria for this analysis. RESULTS: The average weighted reductions in SBP over 8-12 weeks were most marked with diuretics, and in particular indapamide SR 1.5 mg/day (mean change from baseline 22.2mm Hg), which reduced SBP to a greater extent than any of the other drugs evaluated (at any dosage considered). Average weighted reductions in DBP were generally similar with all classes of antihypertensives and ranged from -11.4mm Hg with the beta-adrenoceptor blocker atenolol and calcium channel antagonists to -10.3mm Hg with the angiotensin II type 1 receptor antagonists. CONCLUSION: This new analysis supports the results of the earlier investigation, in that indapamide SR 1.5 mg/day appeared to be the most effective drug for producing significant reductions in SBP within 8-12 weeks, which is an essential element in optimising cardiovascular prevention among hypertensive patients. The clinical application of these results should take into consideration all the limitations discussed in this analysis. PMID- 17914894 TI - Economic evaluation of third-line treatment with alemtuzumab for chronic lymphocytic leukaemia. AB - OBJECTIVE: The objective of this study was to compare the potential economic efficiency of third-line treatment of chronic lymphocytic leukaemia (CLL) with alemtuzumab versus fludarabine, cyclophosphamide and rituximab (FCR). METHODS: The target population for this study were patients with CLL who were able to tolerate third-line treatment with either FCR or alemtuzumab. The perspective used was that of the New Zealand Pharmaceutical Management Agency (PHARMAC)/District Health Board. Health outcomes considered were survival time from commencement of treatment and quality-adjusted life-years (QALYs) gained. Average costs and outcomes and incremental cost per patient treated, per survival month and per QALY gained, were calculated. All costs were presented in 2006 New Zealand dollars. RESULTS: Base-case direct medical costs for alemtuzumab per treatment regimen per patient were $NZ15 303 lower than those for FCR. The average direct medical cost per survival month gained for alemtuzumab was $NZ3144 and for FCR was $NZ4101, and the average direct medical cost per QALY gained was $NZ46,016 and for FCR was $NZ60,012. CONCLUSION: Third-line treatment of eligible patients with alemtuzumab was found to be $NZ15,303 less costly than FCR per patient. PMID- 17914895 TI - Acceptability of lansoprazole orally disintegrating tablets in patients with gastro-oesophageal reflux disease : ACEPTO study. AB - OBJECTIVE: To assess the acceptability of lansoprazole orally disintegrating tablets (LODT) in patients with gastro-oesophageal reflux disease (GORD). METHODS: A multicentre, observational, cross-sectional study of patients diagnosed with GORD aged > or =18 years under the care of 272 gastroenterologists. Acceptability was determined by global patient assessment whereby the drug's organoleptic characteristics and properties were evaluated by a self-administered 11-item ad hoc questionnaire with a 5-point Likert-type scale. RESULTS: A total of 734 patients (mean age 49.6 years [SD = 15.2]) with GORD who had been prescribed LODT > or =14 days prior to inclusion in the study were evaluable for the main endpoint. Of these, 51.1% were men. Most patients (80.7%) had been treated with doses of LODT 30mg/day for an average of 52.7 days (SD = 59.3). Overall, 93.6% of patients rated LODT treatment as 'very acceptable' or ''acceptable'. The degree of acceptability was associated with the perception that the formulation helps treatment compliance (p < 0.001). The drug's properties were rated as follows: size 'neither large nor small' (70.0%); flavour 'very pleasant' or 'pleasant' (75.2%); intensity of flavour 'neither strong nor mild', 'mild' or 'very mild' (86.1%); no 'sandy sensation' (53.4%); speed of dissolving 'fast' or 'very fast' (80.2%); use of tablets 'very easy' or 'easy' (92.4%) and use of tablets 'very convenient' or 'convenient' (91.0%). Three adverse reactions, none of them serious, were reported in three patients (0.4%). CONCLUSIONS: LODT were well accepted by patients with GORD. Patients reported that this formulation improved compliance with therapy. Tolerability was excellent. PMID- 17914896 TI - A registry of the aetiology and costs of neuropathic pain in pain clinics : results of the registry of aetiologies and costs (REC) in neuropathic pain disorders study. AB - OBJECTIVE: The aim of this study was to determine health resource consumption and costs in patients with neuropathic pain managed in pain clinics in Spain. METHODS: This was a retrospective, cross-sectional study performed in 2004 in 18 pain clinics across Spain. Consecutive neuropathic pain patients were recruited between April and December 2004. Demographic data, type and cause of neuropathic pain, source of referral and utilisation of health resources (treatments, medical visits, hospital admissions, etc.) were collected. Direct medical costs were estimated using 2004 prices. Descriptive statistics and ANCOVA models were used for significance. RESULTS: The study included 504 patients with neuropathic pain of broad aetiologies (44% radiculopathies, 21% neuralgias, 11% neuropathies, 7% entrapment syndromes, 5% complex regional painful syndrome, 4% central pain), aged 57.8 +/- 0.7 years (mean +/- SE), 57.6% of whom were women. The mean time since diagnosis was 23.7 +/- 26.8 months. Two groups of patients according to type of pain management were also identified: those referred to pain clinics for pain control from primary-care/other specialists (r-PC, n = 326) and those primarily managed at pain clinics (p-PC, n = 178). The adjusted mean monthly total cost was 363 euros per patient with no statistically significant differences between type of care: 376 euros (p-PC) versus 344 euros (r-PC) [p = 0.626]. Acquisition monthly mean drug costs were higher in the p-PC group: 131 euros (34.8% of total costs) versus 80 euros (23.3%) per patient (p = 0.0001). However, emergency-room visits, primary-care visits and number of treatment drugs were significantly higher in the r-PC group: 0.27, 0.20 and 2.90 visits/drugs per patient per month versus 0.13, 0.10 and 2.50, respectively (p < 0.01 in all cases). There were also more diagnostic tests and specialised physician visits and higher hospitalisation costs in r-PC subjects (differences not statistically significant). CONCLUSIONS: Neuropathic pain results in a substantial utilisation of health resources, particularly by patients referred by primary-care/other specialists to pain clinics for pain control. However, compared with subjects whose pain is primarily managed in pain clinics, the extra health costs arising from drug acquisition observed in such patients are offset by lower costs of the other components of pain management, producing similar mean monthly total costs. PMID- 17914897 TI - BP goal achievement in patients with uncontrolled hypertension : results of the treat-to-target post-marketing survey with irbesartan. AB - BACKGROUND AND OBJECTIVE: Despite the fact that high BP is a leading risk factor for cardiovascular morbidity and mortality, BP goals are achieved in less than 10 30% of hypertensive patients. Irbesartan alone or in combination with hydrochlorothiazide has been shown to control BP in >70% of hypertensive patients in clinical trials. We set out to investigate the role in clinical practice of irbesartan in improving BP in uncontrolled hypertensive patients with a particular focus on patients with the metabolic syndrome through analysis of data from a post-marketing surveillance study. METHODS: A multicentre, prospective, post-marketing surveillance study was conducted over 9 months in 14 200 patients aged > or =18 years with previously uncontrolled hypertension (either receiving therapy or newly diagnosed), paying particular attention to a subgroup of patients receiving irbesartan/hydrochlorothiazide as first-line combination therapy. BP was measured by a sphygmomanometer. The main outcome measures were systolic BP (SBP) and diastolic BP (DBP) reduction, response rate (DBP reduction of > or =10mm Hg or to <90 mm Hg), and BP normalisation (SBP <140 and DBP <90 mm Hg) in patients treated with irbesartan alone or in combination with hydrochlorothiazide. Analyses per patient subgroup, previous medication and whether treatment was initiated by the treating physician as first-line combination therapy were conducted. The number and nature of adverse events were documented. RESULTS: Use of irbesartan 300 mg/day as monotherapy in previously uncontrolled patients resulted in a significant reduction in SBP/DBP (-26.8/ 13.3mm Hg, p < 0.0001), which was comparable to the subgroup of patients with the metabolic syndrome (-26.3/-13.0mm Hg, p < 0.0001 vs baseline). Combination therapy (irbesartan 300 mg/hydrochlorothiazide 12.5mg once daily) lowered BP by 27.9/-14.2mm Hg (p < 0.0001) in previously uncontrolled patients; again the subgroup of patients with the metabolic syndrome achieved a comparable BP reduction (-27.5/-14.1mm Hg, p < 0.0001 vs baseline). Overall, no linear dose response relationship was observed. Use of irbesartan/hydrochlorothiazide as first-line combination therapy was effective (BP normalisation rates between 65.7% and 78.6%) and safe. The mean number of antihypertensive tablets taken was reduced and after a mean period of 9 months, 92% of patients were still taking irbesartan therapy. CONCLUSION: The study demonstrates that treatment with an irbesartan-based regimen for 9 months results in a strong BP reduction and is feasible as first-line combination therapy. Similar BP reductions were observed in the subgroup of patients with the metabolic syndrome. Compliance with treatment is particularly good, with >90% of patients continuing with treatment after 9 months. PMID- 17914898 TI - Effect of gender on the tolerability, safety and pharmacokinetics of clazosentan following long-term infusion. AB - OBJECTIVE: The purpose of this study was to investigate the effect of gender on the tolerability, safety and pharmacokinetics of clazosentan, an intravenous endothelin receptor antagonist. METHODS: Clazosentan was infused at a dosage of 0.05 mg/kg/h or 0.1 mg/kg/h for 72 hours in 8 female and 8 male healthy volunteers, respectively. Tolerability and safety were assessed via the recording of dose reductions/infusion stops, vital signs, ECG, adverse events and clinical laboratory variables. Blood samples were collected for the determination of clazosentan and endothelin-1 concentrations. RESULTS: The occurrence of adverse events such as headache, vomiting and nausea of moderate to severe intensity led either to discontinuation of the infusion or to a dose reduction in the majority of volunteers. The values (mean and 95% confidence intervals) for clearance were 37.7 (32.8, 43.4) and 35.2 (27.8, 44.5) L/h in male and female volunteers, respectively. CONCLUSION: Long-term infusion of clazosentan at the doses tested was poorly tolerated in both male and female volunteers. Gender appeared to have no influence on the pharmacokinetics of clazosentan. PMID- 17914899 TI - Effect of L-carnitine supplementation on endurance exercise in normobaric/normoxic and hypobaric/hypoxic conditions. AB - OBJECTIVE: To evaluate the effect of L-carnitine supplementation on improving endurance exercise in normobaric/normoxic and hypobaric/hypoxic environments. METHODS: Six-week-endurance-trained male Sprague-Dawley rats (n = 24) were randomly divided into 2 groups: control and experimental; the latter group was supplemented with L-carnitine, administered orally in a dose of 100 mg x kg(-1) body weight. The animals were supplemented for 25 days under ambient normobaric/normoxic conditions and thereafter were exposed to 72 hours of hypobaric hypoxia equivalent to 6100 m. The supplementation was continued during the exposure. "Run to exhaustion" was recorded on day 1 (R1) (presupplementation) and on days 7 (R2), 14 (R3), 21 (R4), and 28 (R5, which followed the last 72 hours of hypoxic exposure) of supplementation. Food intake, body weight, and the biochemical measures of plasma glucose, total cholesterol, and high-density lipoprotein (HDL) cholesterol were recorded. RESULTS: There was a significant improvement in endurance exercise, as indicated by an increase in run to exhaustion following L-carnitine supplementation under normobaric normoxia (36% 39%) and hypobaric hypoxia (50%). L-carnitine supplementation had no effect on plasma glucose levels either at sea level or after hypoxic exposure. Total cholesterol was decreased in normoxic and HDL cholesterol was increased in normoxic and hypoxic conditions, indicating a beneficial effect of exercise. CONCLUSION: L-carnitine supplementation improved exercise endurance in rats exposed to normobaric normoxic and hypobaric hypoxic conditions. Such supplementation would be beneficial in delaying the onset of fatigue during prolonged exercise in both conditions, indicating its potentially beneficial use at high altitude. PMID- 17914900 TI - Relationship between mitochondrial electron transport chain dysfunction, development, and life extension in Caenorhabditis elegans. AB - Prior studies have shown that disruption of mitochondrial electron transport chain (ETC) function in the nematode Caenorhabditis elegans can result in life extension. Counter to these findings, many mutations that disrupt ETC function in humans are known to be pathologically life-shortening. In this study, we have undertaken the first formal investigation of the role of partial mitochondrial ETC inhibition and its contribution to the life-extension phenotype of C. elegans. We have developed a novel RNA interference (RNAi) dilution strategy to incrementally reduce the expression level of five genes encoding mitochondrial proteins in C. elegans: atp-3, nuo-2, isp-1, cco-1, and frataxin (frh-1). We observed that each RNAi treatment led to marked alterations in multiple ETC components. Using this dilution technique, we observed a consistent, three-phase lifespan response to increasingly greater inhibition by RNAi: at low levels of inhibition, there was no response, then as inhibition increased, lifespan responded by monotonically lengthening. Finally, at the highest levels of RNAi inhibition, lifespan began to shorten. Indirect measurements of whole-animal oxidative stress showed no correlation with life extension. Instead, larval development, fertility, and adult size all became coordinately affected at the same point at which lifespan began to increase. We show that a specific signal, initiated during the L3/L4 larval stage of development, is sufficient for initiating mitochondrial dysfunction-dependent life extension in C. elegans. This stage of development is characterized by the last somatic cell divisions normally undertaken by C. elegans and also by massive mitochondrial DNA expansion. The coordinate effects of mitochondrial dysfunction on several cell cycle-dependent phenotypes, coupled with recent findings directly linking cell cycle progression with mitochondrial activity in C. elegans, lead us to propose that cell cycle checkpoint control plays a key role in specifying longevity of mitochondrial mutants. PMID- 17914901 TI - MSN2 and MSN4 link calorie restriction and TOR to sirtuin-mediated lifespan extension in Saccharomyces cerevisiae. AB - Calorie restriction (CR) robustly extends the lifespan of numerous species. In the yeast Saccharomyces cerevisiae, CR has been proposed to extend lifespan by boosting the activity of sirtuin deacetylases, thereby suppressing the formation of toxic repetitive ribosomal DNA (rDNA) circles. An alternative theory is that CR works by suppressing the TOR (target of rapamycin) signaling pathway, which extends lifespan via mechanisms that are unknown but thought to be independent of sirtuins. Here we show that TOR inhibition extends lifespan by the same mechanism as CR: by increasing Sir2p activity and stabilizing the rDNA locus. Further, we show that rDNA stabilization and lifespan extension by both CR and TOR signaling is due to the relocalization of the transcription factors Msn2p and Msn4p from the cytoplasm to the nucleus, where they increase expression of the nicotinamidase gene PNC1. These findings suggest that TOR and sirtuins may be part of the same longevity pathway in higher organisms, and that they may promote genomic stability during aging. PMID- 17914902 TI - Nicotinamide riboside kinase structures reveal new pathways to NAD+. AB - The eukaryotic nicotinamide riboside kinase (Nrk) pathway, which is induced in response to nerve damage and promotes replicative life span in yeast, converts nicotinamide riboside to nicotinamide adenine dinucleotide (NAD+) by phosphorylation and adenylylation. Crystal structures of human Nrk1 bound to nucleoside and nucleotide substrates and products revealed an enzyme structurally similar to Rossmann fold metabolite kinases and allowed the identification of active site residues, which were shown to be essential for human Nrk1 and Nrk2 activity in vivo. Although the structures account for the 500-fold discrimination between nicotinamide riboside and pyrimidine nucleosides, no enzyme feature was identified to recognize the distinctive carboxamide group of nicotinamide riboside. Indeed, nicotinic acid riboside is a specific substrate of human Nrk enzymes and is utilized in yeast in a novel biosynthetic pathway that depends on Nrk and NAD+ synthetase. Additionally, nicotinic acid riboside is utilized in vivo by Urh1, Pnp1, and Preiss-Handler salvage. Thus, crystal structures of Nrk1 led to the identification of new pathways to NAD+. PMID- 17914905 TI - Viral evolution in the genomic age. PMID- 17914903 TI - Segregation of odor identity and intensity during odor discrimination in Drosophila mushroom body. AB - Molecular and cellular studies have begun to unravel a neurobiological basis of olfactory processing, which appears conserved among vertebrate and invertebrate species. Studies have shown clearly that experience-dependent coding of odor identity occurs in "associative" olfactory centers (the piriform cortex in mammals and the mushroom body [MB] in insects). What remains unclear, however, is whether associative centers also mediate innate (spontaneous) odor discrimination and how ongoing experience modifies odor discrimination. Here we show in naive flies that Galphaq-mediated signaling in MB modulates spontaneous discrimination of odor identity but not odor intensity (concentration). In contrast, experience dependent modification (conditioning) of both odor identity and intensity occurs in MB exclusively via Galphas-mediated signaling. Our data suggest that spontaneous responses to odor identity and odor intensity discrimination are segregated at the MB level, and neural activity from MB further modulates olfactory processing by experience-independent Galphaq-dependent encoding of odor identity and by experience-induced Galphas-dependent encoding of odor intensity and identity. PMID- 17914906 TI - Can large-scale analysis of the proteome identify effective new markers for embryonic stem cells? PMID- 17914904 TI - Force-induced unfolding of fibronectin in the extracellular matrix of living cells. AB - Whether mechanically unfolded fibronectin (Fn) is present within native extracellular matrix fibrils is controversial. Fn extensibility under the influence of cell traction forces has been proposed to originate either from the force-induced lengthening of an initially compact, folded quaternary structure as is found in solution (quaternary structure model, where the dimeric arms of Fn cross each other), or from the force-induced unfolding of type III modules (unfolding model). Clarification of this issue is central to our understanding of the structural arrangement of Fn within fibrils, the mechanism of fibrillogenesis, and whether cryptic sites, which are exposed by partial protein unfolding, can be exposed by cell-derived force. In order to differentiate between these two models, two fluorescence resonance energy transfer schemes to label plasma Fn were applied, with sensitivity to either compact-to-extended conformation (arm separation) without loss of secondary structure or compact-to unfolded conformation. Fluorescence resonance energy transfer studies revealed that a significant fraction of fibrillar Fn within a three-dimensional human fibroblast matrix is partially unfolded. Complete relaxation of Fn fibrils led to a refolding of Fn. The compactly folded quaternary structure with crossed Fn arms, however, was never detected within extracellular matrix fibrils. We conclude that the resting state of Fn fibrils does not contain Fn molecules with crossed-over arms, and that the several-fold extensibility of Fn fibrils involves the unfolding of type III modules. This could imply that Fn might play a significant role in mechanotransduction processes. PMID- 17914907 TI - HIV/AIDS spread among women. PMID- 17914908 TI - Lyme disease: a turning point. PMID- 17914911 TI - Highlights of the third international workshop on HIV and hepatitis coinfection. PMID- 17914912 TI - Topical triple-antibiotic ointment as a novel therapeutic choice in wound management and infection prevention: a practical perspective. AB - Triple-antibiotic ointment (TAO) is a safe and effective topical agent for preventing infections in minor skin trauma. The formulation contains neomycin, polymyxin B and bacitracin in a petrolatum base. TAO is active against the most common disease-causing pathogens found in wounds and on the skin and may be an attractive alternative to oral therapy in select circumstances. Resistance to TAO does not develop readily, and safety studies have shown that the risk of allergic sensitivity to TAO is low. Susceptibility profiles of TAO have remained relatively unchanged since its discovery. Prophylaxis or treatment with TAO should be considered as resistant organisms continue to emerge in the community and hospital setting. PMID- 17914913 TI - Potential use of inhibitors of bacteria spore germination in the prophylactic treatment of anthrax and Clostridium difficile-associated disease. AB - Spore germination is the first step in establishing Bacillus and Clostridium infections. Germination is triggered by the binding of small molecules by the resting spore. Subsequently, the activated spore secretes dipicolinic acid and calcium, the spore core is rehydrated and spore structures are degraded. Inhibition of any of the germination-related events will prevent development to the vegetative stage. Inhibition of spore germination has been studied intensively in the prevention of food spoilage. In this perspective, we propose that similar approaches could be used in the prophylactic control of Bacillus anthracis and Clostridium difficile infections. Inhibition of B. anthracis spore germination could protect military and first-line emergency personnel at high risk for anthrax exposure. Inhibition of C. difficile could prevent human C. difficile-associated disease during antibiotic treatment of immunocompromised patients. PMID- 17914914 TI - Carbapenems in the USA: focus on doripenem. AB - The threat of antibacterial resistance continues to increase globally, and therapeutic options for the treatment of some serious infectious diseases are diminishing. The carbapenems are a potent class of broad-spectrum drugs, and their stability against hydrolysis by many important beta-lactamases make them an important weapon in the treatment of beta-lactamase-producing bacterial pathogens. This review focuses on four carbapenems of clinical importance in the USA: imipenem, meropenem, ertapenem and doripenem. After a historical review of carbapenem development, these four carbapenems are evaluated based on their mechanism of action, spectrum of activity, potency, pharmacodynamics, clinical pharmacokinetics, clinical profiles and toxicity issues. PMID- 17914915 TI - Polymyxin B: similarities to and differences from colistin (polymyxin E). AB - Hospital-acquired infections due to multidrug-resistant gram-negative bacteria constitute major health problems, since the medical community is continuously running out of available effective antibiotics and no new agents are in the pipeline. Polymyxins, a group of antibacterials that were discovered during the late 1940s, represent some of the last treatment options for these infections. Only two polymyxins are available commercially, polymyxin E (colistin) and polymyxin B. Although several reviews have been published recently regarding colistin, no review has focused on the similarities and differences between polymyxin B and colistin. These two medications have many similarities with respect to mechanism of action, antimicrobial spectrum, clinical uses and toxicity. However, they also differ in several aspects, including chemical structure, formulation, potency, dosage and pharmacokinetic properties. PMID- 17914916 TI - Potential role of RNAi in the treatment of HCV infection. AB - Chronic HCV infection is a leading cause of chronic hepatitis and its sequelae, liver cirrhosis and hepatocellular carcinoma. Current therapeutic options are limited, associated with significant adverse effects and costly. Accordingly, there is strong impetus to develop novel therapeutic strategies that act through alternate mechanisms. RNAi has been widely used for the analysis of gene function and represents a potentially promising approach for the treatment of HCV infection. siRNAs are short RNA duplexes approximately 21 nts long. When introduced into mammalian cells, siRNA can silence specific gene expression. Although efficient suppression of HCV replicon RNA in cell culture has been demonstrated with siRNAs, there is much work to be done to improve delivery, limit off-target effects and minimize development of virus resistance. Here, we review the use of RNAi as a tool to inhibit HCV gene expression and discuss the potential advantages and obstacles for this new potential therapeutic approach against HCV infection. PMID- 17914917 TI - Current concepts in antibiotic-resistant gram-negative bacteria. AB - Gram-negative bacteria are the dominant killers among bacterial pathogens in the intensive care unit. Antibiotic resistance has become a threat in hospital settings and efforts are being made to understand the underlying mechanisms. This review describes current data on the most important mechanisms of resistance in prevalent gram-negative pathogens as well as newer therapeutic options. PMID- 17914918 TI - Current management of Helicobacter pylori infections in the elderly. AB - Helicobacter pylori infection is a chronic gastric gram-negative infection that increases with age worldwide. However, the percentage age of H. pylori-positive elderly patients who are tested and treated for their infection remains very low. It is now demonstrated that H. pylori infection induces a whole cascade of events leading to gastric pathologies, such as peptic ulcer diseases, gastric precancerous lesions and gastric cancer. Recent data also demonstrated that H. pylori chronic infection can play a role in gastric aging, appetite regulation and extradigestive diseases, such as Alzheimer's disease, in the elderly. The diagnosis of H. pylori infection remains difficult to realize in the very old population, and the urea breath test obtains the best performance in this population. 1-week proton pump inhibitor-based triple therapy regimens are highly effective and well tolerated in elderly patients, and antibiotic resistance remains very low. Low compliance is the main factor related to treatment failure in this population. PMID- 17914919 TI - Multidrug-resistant and extensively drug-resistant Mycobacterium tuberculosis: epidemiology and control. AB - The emergence of multidrug-resistant (MDR)-TB and, more recently, of extensively drug-resistant (XDR)-TB is a real threat to achieve TB control and elimination. Over 400,000 new cases of MDR-TB occur each year and, although their number is currently unknown, XDR cases are recognized in every setting where there has been the capacity to detect them. The long-term vision for the full control of MDR-TB requires the scaling-up of culture and drug-susceptibility testing capacity, which is very limited in disease-endemic countries, and the expanded use of high technology assays for rapid determination of resistance. MDR cases are treatable and well designed regimens, largely based on second-line anti-TB drugs, can considerably improve cure rates. However, treatment regimens need to be markedly improved through the introduction of less toxic and more powerful drugs, thus reducing duration of treatment and tolerability. This is of utmost importance for XDR-TB cases. The prevalence of MDR-TB and XDR-TB are inversely correlated with the quality of TB control and the proper use of second-line anti-TB drugs. Adherence to proper standards of care and control is imperative and a top priority of all TB control efforts. However, the risk of an uncontrollable epidemic of MDR- and XDR-TB is real considering current levels of financing and commitment to care. PMID- 17914921 TI - Combination antifungals: an update. AB - Invasive fungal infections are an important cause of morbidity and mortality in specific patient populations. There has been an impressive increase in the antifungal armamentarium, yet optimal therapies for many invasive fungal infections remain unknown. Genomic sequencing of a number of pathogenic fungi will pave the way to discovering additional newer targets for antifungal drug design. These new discoveries, plus the existing repertoire of antifungal agents, create the need to effectively model single and combination antifungal agents. Future therapies may also include the use of cell-stress pathway inhibitors in combination with existing antifungal agents. This review focuses on combination antifungal therapy against Cryptococcus neoformans, Candida and Aspergillus species. Combination therapy is only supported by randomized clinical trials for cryptococcal meningitis. We review data from in vitro and animal model studies as well as insights from clinical trials to discuss current thoughts and highlight the gaps in our knowledge surrounding combination antifungal therapy. PMID- 17914920 TI - Antibody response to Salmonella: its induction and role in protection against avian enteric salmonellosis. AB - Human enteritis resulting from the consumption of poultry products contaminated with serovars of Salmonella enterica remains a major public-health concern. Reducing food contamination by preventing or controlling infection in the chicken during rearing is an attractive solution. An accurate understanding of the mechanisms of immunity to Salmonella infection in the chicken will help to focus the development of vaccines for birds and prevent contaminated products from entering the human food chain. Infection is primarily restricted to the intestinal lumen when chickens are infected with S. enterica serovars Typhimurium or Enteritidis, where they persist for many weeks. High titers of Salmonella specific antibodies are observed following infection and demonstrate a high degree of cross-reactivity against other serovars. However, depletion of B cells and, therefore, removal of the capacity for antibody production in the chicken does not exacerbate the infection following either primary or secondary challenge. PMID- 17914922 TI - Invasive amebiasis: an update on diagnosis and management. AB - In its invasive form, the trophozoite is responsible for clinical syndromes, ranging from classical dysentery to extraintestinal disease with emphasis on hepatic amebiasis. Abdominal pain, tenderness and diarrhea of watery stool, sometimes with blood, are the predominant symptoms of amebic colitis. Besides the microscopic identification of Entamoeba histolytica, diagnosis should be based on the detection of specific antigens in the stool or PCR associated with the occult blood in the stool. Amebic dysentery is treated with metronidazole, followed by a luminal amebicide. The trophozoite reaches the liver causing hepatic amebiasis. Right upper quadrant pain, fever and hepatomegaly are the predominant symptoms. The diagnosis is made by the finding of E. histolytica in the hepatic fluid, or in the necrotic material at the edge of the lesion in a minority of patients, and by detection of antigens or DNA. Ultrasonography is the initial imaging procedure indicated. The local perforation of hepatic lesion leads to important and serious complications. PMID- 17914923 TI - Long-term maintenance of neuronally differentiated human adipose tissue-derived stem cells. AB - Recent studies reporting differentiation of early neural progenitors of human adipose tissue-derived stromal cells (ADSCs) has aroused interest among investigators for regenerative medicine. The aim of this study was to investigate the differentiation of ADSCs to neuron-like cells and to extend the life span of these differentiated ADSCs in vitro using our new DE-1 medium. After primary culture and expansion, ADSCs were incubated in a new long-term neuronal induction medium that maintains ADSCs in a differentiated state for 8 weeks. Neuronal differentiation was identified using immunocytochemistry, reverse-transcriptase polymerase chain reaction, and Western blotting. We found that the optimal differentiation protocol induced the ADSCs to express early neuronal markers, including nestin and neuronal nuclear antigen (NeuN), as well as the mature astrocyte marker glial fibrillary acidic protein (GFAP). Neuronal morphological characteristics were recognized in approximately 40% to 50% of the cell populations maintained over 8 weeks, and 60% to 80% of the differentiated cells expressed neuronal specific markers, including nestin, GFAP, NeuN, Trk-A, vimentin, and neuron-specific enolase. The data show that our DE-1 medium is capable of achieving a greater number of differentiated ADSCs for a longer period of time. This result bodes well for the application of ADSCs in in vivo peripheral nerve regeneration. PMID- 17914924 TI - Adaptive treatment strategies in chronic disease. AB - An adaptive treatment strategy (ATS) is a rule for adapting a treatment plan to a patient's history of previous treatments and the response to those treatments. The ongoing management of chronic disease defines an ATS, which may be implicit and hidden or explicit and well-specified. The ATS is characterized by the use of intermediate, early markers of response to dynamically alter treatment decisions, in order to achieve a favorable ultimate outcome. We illustrate the ATS concept and describe how the effect of initial treatment decisions depends on the performance of subsequent decisions at later stages. We show how to compare two or more ATSs, or to determine an optimal ATS, using a sequential multiple assignment randomized (SMAR) trial. Designers of clinical trials might find the ATS concept useful in improving the efficiency and ecological relevance of clinical trials. PMID- 17914925 TI - Genetic determinants of aggressive breast cancer. AB - The development and spread of breast and other human cancers are caused by the overexpression, mutation, and/or deletion of specific genes that drive these events. Finding genetic and molecular differences between cancerous and healthy cells can reveal the genetic determinants of cancer. This knowledge results in a better understanding of the carcinogenic process and improved predictive power, with implications for identifying new drug targets, designing novel therapies, and improving preclinical and clinical studies. We review the concepts of biomarker, genetic marker, and genetic determinant in cancer, with particular focus on the most aggressive and lethal form of breast cancer, termed inflammatory breast cancer (IBC). Using IBC as an example, we describe in detail the approaches to identify the genes that are responsible for-and not merely associated with-this disease. PMID- 17914926 TI - Mesenchymal stem cells in acute kidney injury. AB - The potential role of mesenchymal stem cells (MSCs, also called mesenchymal stromal cells) in endogenous repair and cell-based therapies for acute kidney injury (AKI) is under intensive investigation. Preclinical studies indicate that administered MSCs both ameliorate renal injury and accelerate repair. These versatile cells home to sites of injury, where they modulate the repair process. The mechanisms responsible for their protective and regenerative effects are incompletely understood. Some have reported that MSCs are capable of direct engraftment into injured nephrons under certain circumstances. This is highly controversial, however, and even those who argue there is engraftment acknowledge that the primary means of repair by these cells most likely involves paracrine and endocrine effects, including mitogenic, antiapoptotic, anti-inflammatory, and angiogenic influences. There is a good deal of interest in MSC-based approaches for the treatment of human kidney injury, thanks to positive preclinical results, the strong clinical need for novel therapies to treat AKI, the ease of isolation and expansion of MSCs, and encouraging preliminary clinical trial results in other fields. This review summarizes current knowledge and identifies gaps in our understanding of MSC biology that will need to be filled in order to translate recent discoveries into therapies for AKI in humans. PMID- 17914927 TI - Mechanism-based concepts of size and maturity in pharmacokinetics. AB - Growth and development can be investigated using readily observable demographic factors such as weight and age. Size is the primary covariate and can be referenced to a 70-kg person with allometry using a coefficient of 0.75 for clearance and 1 for volume. The use of these coefficients is supported by fractal geometric concepts and observations from diverse areas in biology. Fat free mass (FFM) might be expected to do better than total body weight when there are wide variations in fat affecting body composition. Clearance pathways develop in the fetus before birth. The use of postnatal age as a descriptor of maturation is unsatisfactory because birth may occur prematurely; therefore postmenstrual age is a superior predictor of elimination function. A sigmoid E(max) model (Hill equation) describes gradual maturation of clearance in early life leading to a mature adult clearance achieved at a later age. PMID- 17914928 TI - Role of CYP1B1 in glaucoma. AB - Glaucoma is a leading cause of blindness, estimated to affect 60 million people by 2010, and represents a heterogeneous group of neurodegenerative disease. The two major types of glaucoma include primary open-angle glaucoma (POAG) and primary congenital glaucoma (PCG). A genetically heterogeneous group of developmental disorders known as anterior segment dysgenesis (ASD) have been reported to be associated with increased intraocular pressure (IOP) and glaucoma. These include Peters' anomaly, Rieger's anomaly, aniridia, iris hypoplasia, and iridogoniodysgenesis. Genetic linkage analysis and mutation studies have identified CYP1B1 as a causative gene in PCG, as a modifier gene in POAG, and, on rare occasions, as causative gene in POAG as well as in several ASD disorders. CYP1B1-deficient mice exhibit abnormalities in their ocular drainage structure and trabecular meshwork that are similar to those reported in human PCG patients. Accordingly, it is speculated that diminished or absent metabolism of key endogenous CYP1B1 substrates adversely affects the development of the trabecular meshwork. CYP1B1 protein is involved in the metabolism of steroids, retinol and retinal, arachidonate, and melatonin. The conserved expression of CYP1B1 in both murine and human eyes, its higher expression in fetal than adult eyes, and its biochemical properties are consistent with this hypothesis. The exact role of CYP1B1 in the pathogenesis of glaucoma and other ASD disorders remains to be elucidated. PMID- 17914929 TI - Pharmacotherapy for the treatment of choroidal neovascularization due to age related macular degeneration. AB - Age-related macular degeneration (AMD) is a progressive, degenerative disease of the macula that threatens central vision. It initially occurs in a "dry" form, and can progress to choroidal neovascularization (CNV) or geographic atrophy. It is the leading cause of blindness among European-descended people older than 65 years, with a prevalence of 1.5%. The treatment of CNV in developed nations in 2007 is substantially different than it was in 1997. Focal, photocoagulating, laser therapy was replaced by intravenous verteporfin and then by intravitreal pegaptanib, which is now being replaced by intravitreal ranibizumab and off-label use of bevacizumab. Other than a ranibizumab versus verteporfin trial, there are no published comparative studies of the three approved pharmacological treatments for CNV. Although frequent intravitreal injections are accepted as a current standard of care, their use is still far from ideal. Thus, there is an opportunity for improving therapy of CNV with respect to mechanism-targeted treatments, efficacy, and route of administration. PMID- 17914931 TI - Protective interventions to prevent aflatoxin-induced carcinogenesis in developing countries. AB - The public health impact of aflatoxin exposure is pervasive in economically developing countries; consequently, we need to design intervention strategies for prevention that are practicable for these high-risk populations. The adverse health consequences of aflatoxins in populations are quite varied, eliciting acute effects, such as rapid death, and chronic outcomes, such as hepatocellular carcinoma. Furthermore, a number of epidemiological studies describe a variety of general adverse health effects associated with aflatoxin, such as impaired growth in children. Thus, the magnitude of the problem is disseminated across the entire spectrum of age, gender, and health status in the population. The aflatoxins multiplicatively increase the risk of liver cancer in people chronically infected with hepatitis B virus (HBV), which illustrates the deleterious impact that even low toxin levels in the diet can pose for human health. Thus other aflatoxin interactions, which likely contribute to the disease burden, still remain to be identified. Therefore, many diverse and appropriate strategies for disease prevention are needed to decrease the incidence of aflatoxin carcinogenesis in developing countries. PMID- 17914932 TI - A critical review of theory in breast cancer screening promotion across cultures. AB - This article reviews the contribution and potential of widely used health behavior theories in research designed to understand and redress the disproportionate burden of breast cancer borne by diverse race/ethnic, immigrant, and low-income groups associated with unequal use of mammography. We review the strengths and limitations of widely used theories and the extent to which theory contributes to the understanding of screening disparities and informs effective intervention. The dominant focus of most theories on individual cognition is critically assessed as the abstraction of behavior from its social context. Proposed alternatives emphasize multilevel ecological approaches and the use of anthropologic theory and methods for more culturally grounded understandings of screening behavior. Common and alternative treatments of fatalism exemplify this approach, and descriptive and intervention research exemplars further highlight the integration of screening behavior and sociocultural context. PMID- 17914933 TI - Ecologic studies revisited. AB - Ecologic studies use data aggregated over groups rather than data on individuals. Such studies are popular because they use existing databases and can offer large exposure variation if the data arise from broad geographical areas. Unfortunately, the aggregation of data that define ecologic studies results in an information loss that can lead to ecologic bias. Specifically, ecologic bias arises from the inability of ecologic data to characterize within-area variability in exposures and confounders. We describe in detail particular forms of ecologic bias so that their potential impact on any particular study may be assessed. The only way to overcome such bias, while avoiding uncheckable assumptions concerning the missing information, is to supplement the ecologic with individual-level information, and we outline a number of proposals that may achieve this aim. PMID- 17914934 TI - Deja vu, mais pas en anglais! Precautionary notes on publishing the same article in two languages. PMID- 17914930 TI - Caveolae as organizers of pharmacologically relevant signal transduction molecules. AB - Caveolae, a subset of membrane (lipid) rafts, are flask-like invaginations of the plasma membrane that contain caveolin proteins, which serve as organizing centers for cellular signal transduction. Caveolins (-1, -2, and -3) have cytoplasmic N and C termini, palmitolylation sites, and a scaffolding domain that facilitates interaction and organization of signaling molecules so as to help provide coordinated and efficient signal transduction. Such signaling components include upstream entities (e.g., G protein-coupled receptors (GPCRs), receptor tyrosine kinases, and steroid hormone receptors) and downstream components (e.g., heterotrimeric and low-molecular-weight G proteins, effector enzymes, and ion channels). Diseases associated with aberrant signaling may result in altered localization or expression of signaling proteins in caveolae. Caveolin-knockout mice have numerous abnormalities, some of which may reflect the impact of total body knockout throughout the life span. This review provides a general overview of caveolins and caveolae, signaling molecules that localize to caveolae, the role of caveolae/caveolin in cardiac and pulmonary pathophysiology, pharmacologic implications of caveolar localization of signaling molecules, and the possibility that caveolae might serve as a therapeutic target. PMID- 17914935 TI - Magnifying chromoendoscopy: effective diagnostic tool for screening colonoscopy. PMID- 17914936 TI - Do serrated adenomas have higher malignant potential than traditional adenomas? PMID- 17914937 TI - Iron and hepatitis C: beginning to make sense. PMID- 17914938 TI - Resection for hepatic hemangiomas. PMID- 17914939 TI - Endoscopic diagnoses and CLO test results in 9239 cases, prevalence of Helicobacter pylori in Istanbul, Turkey. AB - BACKGROUND: Helicobacter pylori is implicated in the etiology of gastric and duodenal ulcer, non-ulcer dispepsia, atrophic gastritis, gastric adenocarcinoma and lymphoma. METHODS: Between November 1995 and December 2004, the presence of H. pylori was investigated using the CLO test in 9239 patients who underwent upper gastrointestinal endoscopy at a single institution in Istanbul, Turkey. The results were evaluated as early-late positive, and negative. RESULTS: There were 4667 women (50.51%) with a mean age of 44.5 years (range, 13-90 years), and 4572 men (49.49%) with a mean age of 45.7 years (range, 11-85 years). The CLO test was positive in 41.44% of cases. The most frequent symptoms on admission were epigastric pain (46.2%) and burning (19.6%). The most frequent endoscopic diagnosis was pangastritis (64%) and non-erosive duodenitis (30.5%). The H. pylori positivity was 61.53% during the first 5-year period and 38.47% during the second 5-year period. The H. pylori positivity was significant in patients using non-steroidal anti-inflamatory drugs and tobacco (P < 0.001). DISCUSSION AND CONCLUSION: Helicbacter pylori prevelance remains an important health problem for Turkey although it has diminished in parallel to the national development during the last years. Helicbacter pylori, as a first-degree carcinogen, should be investigated and eradicated particularly in high-risk patients. PMID- 17914940 TI - Utility of endoscopic ultrasound in the diagnosis of aberrant right subclavian artery. AB - BACKGROUND AND AIMS: Aberrant right subclavian artery (ARSA) is the most common congenital anomaly of the aortic arch occurring in 0.4-2.0% of the general population. Instead of arising from the brachiocephalic artery, the ARSA arises as the last branch from the aortic arch. The aim of this study was to determine the prevalence of ARSA in patients undergoing upper endoscopic ultrasound (EUS) and to describe the EUS characteristics of ARSA. METHODS: A retrospective review was conducted of 7513 upper EUS exams performed from 1 July 2000 to 1 February 2005. RESULTS: In total, 27 (0.36%, 95% CI 0.22-0.50%) of 7513 patients undergoing upper EUS were found to have an ARSA (10 male, 17 female; mean age 58 years, range 23-81 years). Of the 27 patients, 16 had only radial EUS, 10 had radial and linear EUS, and one had only linear EUS. In all 26 patients who underwent radial imaging, a well-defined, anechoic tubular structure was seen originating from the aortic arch and passing between the esophagus and spine. In all 11 patients undergoing linear imaging, the abnormal anatomy was detected and the vascular nature confirmed by Doppler. In one patient, both radial and linear imaging identified unsuspected aneurismal dilatation of the ARSA; a rare condition referred to as Kommerell's diverticulum (KD). None of the other 26 patients had symptoms to suggest an ARSA. Of the 14 patients who had computed tomography (CT) prior to EUS, only four were initially reported to have ARSA. However, later review of the CT scans verified an ARSA in all 14 patients. Of the two patients who underwent a barium swallow, only one had findings suggestive of ARSA. CONCLUSIONS: This report highlights the utility of both radial and linear EUS imaging in identifying previously unsuspected ARSA and associated anomalies. This report also suggests the need for careful review of the CT in patients suspected of having ARSA due to the frequent failure of radiologists to identify, or report, this anomaly when particular focus is not given. PMID- 17914941 TI - Association of body mass index with heartburn, regurgitation and esophagitis: results of the Progression of Gastroesophageal Reflux Disease study. AB - BACKGROUND: Overweight and obesity are believed to be risk factors for gastroesophageal reflux disease (GERD). The aim of the present study was to analyze the impact of body mass index (BMI) on the severity and frequency of reflux symptoms and esophagitis in a large cohort of reflux patients. METHODS: As part of the Progression of Gastroesophageal Reflux Disease (ProGERD) study, 6215 patients with clinically assessed GERD were included in the present investigation (53% male, 52 +/- 14 years; 47% female, 56 +/- 14 years). Heartburn and regurgitation symptoms were assessed using the validated Reflux Disease Questionnaire. Endoscopies were performed and patients were subsequently classified as having non-erosive or erosive disease. To examine the association between BMI, GERD symptoms, and esophagitis, odds ratios (OR) and 95% confidence intervals (95%CI) were calculated using logistic regression models. RESULTS: In patients with GERD, higher BMI was associated with more frequent and more severe heartburn and regurgitation, as well as with esophagitis. The effects were more pronounced for regurgitation than for heartburn. The strongest association was between obesity and severity of regurgitation symptoms (women: OR 2.11, 95%CI 1.60-2.77; men: OR 2.15, 95%CI 1.59-2.90). Obese women, but not men, had an increased risk of severe esophagitis compared to women with normal weight (OR 2.51, 95%CI 1.53-4.12). CONCLUSIONS: In patients with GERD, higher BMI was associated with more severe and more frequent reflux symptoms and esophagitis. PMID- 17914942 TI - Study of arteriosclerosis in patients with hiatal hernia and reflux esophagitis. AB - BACKGROUND AND AIM: It has been reported that the prevalence of hiatal hernia (HH) and reflux esophagitis (RE) increases with age, as does the degree of arteriosclerosis. However, it has not been investigated whether or not arteriosclerosis is correlated with the presence of HH and RE. Therefore, we prospectively investigated the degree of arteriosclerosis in patients with HH and RE compared with subjects without HH and RE. METHODS: We prospectively enrolled 1683 people who visited Shimane Environment and Health Public Corporation for annual medical check-ups. All subjects were investigated by upper gastrointestinal endoscopy for the possible presence of HH and RE. Factors used for assessing cardiovascular risk were sex, age, body mass index (BMI), smoking and drinking habits, serum total cholesterol, triglyceride (TG), and high-density lipoprotein cholesterol (HDLC). Arteriosclerosis was investigated by measuring systolic blood pressure, heart-carotid pulse wave velocity (HCPWV), bilateral ankle brachial index (ABI) and heart-ankle PWV (HAPWV). RESULTS: The number of patients with HH and RE was 624 (37.1%) and 143 (8.5%), respectively. The HDLC level of the patients with HH was significantly lower, and the levels of BMI, TG and arteriosclerotic parameters were higher than those of the subjects without HH after adjusting for confounding factors (sex, age and smoking and drinking habits). Cardiovascular and arteriosclerosis parameters except for TG did not differ between the subjects with and without RE. CONCLUSION: Levels of arteriosclerosis parameters in patients with HH were higher than in those without HH. However, the association between arteriosclerosis and presence of RE was not clarified. PMID- 17914943 TI - Esophageal motility differences among aged patients with achalasia: a Taiwan report. AB - BACKGROUND AND AIM: There are limited reports on esophageal motility pressures in aged patients with achalasia and these are inconclusive. The aim of the present retrospective study was to understand the changes of esophageal motility in aged achalasia patients among the Taiwan population. METHODS: Manometric studies of 49 patients with achalasia had been performed through January 1998 to June 2005. The findings of lower esophageal sphincter (LES) basal and residual pressures and esophageal body contraction amplitudes were calculated and compared between the older and younger patient groups at different age cut-offs. RESULTS: Higher basal LES pressure increased significantly from the cut-off age of 65 years (i.e. patients over 65 had significantly higher basal LES pressure than younger patients: 37.0 +/- 4.19 mmHg vs 30.0 +/- 1.32 mmHg, P = 0.045). With patients > or =70 years old, it was more obvious (46.0 +/- 3.7 mmHg vs 29.6 +/- 1.2 mmHg, P = 0.001). Beginning at the cut-off age of 55, the LES residual pressure was significantly higher in older patients than those who were younger (14.0 +/- 11.06 mmHg vs 11.1 +/- 0.6 mmHg, P = 0.017). LES residual pressure is more significant in the older groups. A linear correlation between age and residual LES pressures (r = 0.383) was found. No differences were found in esophageal contraction pressure. CONCLUSIONS: Older achalasia patients in Taiwan have higher basal LES pressures, with a linear correlation between age and residual LES pressures. Age has no influence on esophageal contraction pressure. PMID- 17914944 TI - Clinical outcome of rumination syndrome in adults without psychiatric illness: a prospective study. AB - BACKGROUND AND AIM: This prospective study was conducted to characterize the clinical features of adult rumination syndrome, in the absence of psychiatric illness, by applying newly modified clinical criteria, and to elucidate factors influencing outcomes after treatment. METHODS: Twenty-one adult patients diagnosed with rumination syndrome at a tertiary referral center over a 2-year period were enrolled in the study. All patients received supportive psychotherapy and medical treatment. Changes in symptom parameters were analyzed using a pretreatment and a post-treatment questionnaire. Patients were classified into three groups according to symptomatic outcome: improved group, sustained group and aggravated group. RESULTS: The duration of treatment was longer in the improved group than in the sustained or aggravated groups (P = 0.018). Esophageal manometry testing demonstrated non-transmitted contractions of the esophageal body in 10 patients (47.6%) and low amplitude contractions in eight patients (38.1%). Mean lower esophageal sphincter (LES) pressure was 9.8 +/- 2.6 mmHg. Baseline LES pressure in the improved group was higher than in the other groups (P = 0.001). Ambulatory 24-h esophageal pH monitoring showed pathological acid reflux in six patients (28.6%). All pathological acid reflux occurred in post rumination periods. Scintigraphic testing of gastric emptying revealed that the mean retention rate at 120 min was 34.2 +/- 2.5% of initial contents, and delayed emptying was noted in three patients (14.3%). CONCLUSIONS: Rumination syndrome is often accompanied by heterogeneous conditions such as postprandial gastroesophageal reflux, various abnormalities in esophageal manometric tests and delayed gastric emptying. Medical treatment and supportive psychotherapy can be effective in otherwise normal adult patients, especially in patients who comply with long durations of treatment and who demonstrate higher baseline LES pressures. PMID- 17914945 TI - Impact of sleep disturbances in inflammatory bowel disease. AB - BACKGROUND: Normal sleep is paramount for a healthy lifestyle and high quality of life. Sleep modulates the immune system and thus affects the course of several chronic inflammatory conditions. There are no reported studies that address the role of sleep disturbance in the course of inflammatory bowel disease (IBD). The aim of this study was to characterize sleep disturbance in IBD using validated measures of sleep and quality of life. METHODS: A self-administered, mail-in questionnaire package was sent to 205 subjects after a brief instruction. The questionnaire package was composed of the Pittsburgh Sleep Quality Index (PSQI), a measure of disease severity and the IBD-Quality of Life Questionnaire. A total of 119 subjects were recruited (58% response rate): 80 with inactive IBD, 24 with irritable bowel syndrome (IBS) and 15 healthy controls. RESULTS: The IBD subjects reported significantly prolonged sleep latency, frequent sleep fragmentation, higher rate of using sleeping pills, decreased day-time energy, increased tiredness and poor overall sleep quality compared to healthy controls. The abnormal sleep patterns in IBD subjects were similar to IBS subjects. The reported sleep quality was correlated with IBD disease severity score (r(2) = 0.55, P = 0.02). Both IBD and IBS subjects thought that sleep and their disease status were correlated. CONCLUSION: The results show that IBD patients have significant sleep disturbance even when their disease is not active. This problem might affect quality of life, gastrointestinal symptoms and coping ability, and might potentially modify disease severity or increase risk of flare-up. Regardless of the primary or secondary origin of this problem, sleep disturbance should be addressed in the clinical management of patients with IBD. PMID- 17914946 TI - Gastrointestinal stromal tumors: usefulness of immunohistochemistry, flow cytometry and fluorescence in situ hybridization. AB - BACKGROUND AND AIMS: Gastrointestinal stromal tumors (GIST) constitute a group of primary mesenchymal tumors of the gastrointestinal tract, known for their diversity in clinical behavior and the difficulties in determining malignancy and prognosis. This retrospective study evaluated a series of GIST by means of immunohistochemical techniques, flow cytometry and fluorescence in situ hybridization (FISH). METHODS: Nine patients with GIST were analyzed for tumor size, mitotic count and CD117, CD34, MIB-1 with immunohistochemistry. In addition, the GIST were tested with FISH for chromosomes 8 and 17 and DNA index was evaluated by flow cytometry. RESULTS: The findings confirmed the usefulness of CD117 and CD34 in diagnosing GIST and the prognostic role of MIB-1, but do not support a correlation between aneuploidy in flow cytometry and poor outcome. The FISH results suggest close follow-up for patients with benign GIST with a numerical alteration of chromosome 8. The technique could select patients with tumors at high-risk with aneusomy of chromosome 17. CONCLUSION: This study shows the possible application of FISH to the evaluation of patients with GIST, in addition to analysis of morphological features. PMID- 17914947 TI - TLR2, TLR4 and TLR9 polymorphisms and Crohn's disease in a New Zealand Caucasian cohort. AB - BACKGROUND AND AIM: Toll-like receptors (TLRs) have been identified as susceptibility genes for Crohn's disease (CD) in some, but not all, studies. Here we examined the association between candidate disease-susceptibility polymorphisms in the TLR2, TLR4 and TLR9 genes and CD in a New Zealand Caucasian population. METHODS: The frequency of gene polymorphisms was examined in 182 CD patients and in 188 ethnically matched controls by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. RESULTS: We could not detect any significant difference in the allele frequencies of polymorphisms in the TLR2 (R753Q, 0.029 vs 0.016, P = 0.25), TLR4 (D299G and T399I, 0.085 vs 0.071, P = 0.49; and 0.085 vs 0.082, P = 0.90), and TLR9 (-1237T/C, 0.154 vs 0.148, P = 0.82) genes between controls and patients, respectively. There was no evidence that the variant TLR alleles were associated with disease phenotype. However, combination of the datasets of published studies with our dataset confirmed that the TLR4 polymorphism 299G (P = 0.0005; OR of 1.42 [95% CI 1.17 1.74]) and the TLR9 polymorphism -1237C (P = 0.0416; OR of 1.33 [95% CI 1.01 1.75]) are associated with CD. CONCLUSIONS: There was no evidence that the above variants of the TLR2, TLR4 and TLR9 genes are major risk factors for CD or influence disease phenotype in our New Zealand case-control study. Nevertheless, the significance of the TLR4 299G and TLR9-1237C associations with CD worldwide was confirmed by a meta-analysis test using our datasets and datasets from previously published studies. PMID- 17914948 TI - Adult intussusception in Asians: clinical presentations, diagnosis, and treatment. AB - BACKGROUND: Adult intussusception is a rare clinical entity. The presentations and optimal management of adult intussusception in Asians have seldom been reported. The purpose of this study was to determine the causes and management of adult intussusception in Taiwan. METHODS: A retrospective review performed at four medical centers in Taiwan identified 46 patients, at least 18 years old, with a diagnosis of intestinal intussusception from January 1992 to December 2005. Data related to presentations, diagnosis, treatment, and pathology were analyzed. RESULTS: There were 28 men and 18 women with a mean age of 58 (range 19 83) years. Twenty-five patients were diagnosed with enteric intussusception and 21 patients with colonic intussusception. Disease in 35 (76.1%) patients was caused by a benign lead point. The most common symptom was abdominal pain, which was seen in all patients. Twenty-five patients presented with acute symptoms that they experienced over a period of less than 3 days. The preoperative diagnosis was 89.1% because of the wide use of abdominal computed tomography (CT). The most sensitive diagnostic modality was abdominal CT (88.6%). For all patients, 76% with enteric intussusception and 28.6% with colonic intussusception underwent operative reduction. At the time of writing, 71.7% patients were still alive. Only two patients died of postoperative complications. CONCLUSIONS: Most patients with adult intussusception in our series were men, and most intussusceptions were benign and of enteric origin. The most sensitive diagnostic modality is abdominal CT scan. Operative reduction is recommended for enteric intussusceptions but not for colonic intussusceptions. The prognosis of adult intussusception after surgery is good except for malignant intussusception. PMID- 17914949 TI - Infective colitis associated with human intestinal spirochetosis. AB - AIM: Our study reports the detection and identification of intestinal spirochetosis in patients with colonic diseases in a tertiary-care hospital over a 12-year period, and includes a description of all cases we diagnosed. METHODS: Our patients (8323) underwent colonoscopy and histopathological examinations including transmission electron microscopy (TEM) and light microscopy. Specimens from patients suspected of intestinal spirochetosis at histopathology (17 patients) underwent microbiological investigation performed by culture and molecular methods (16S restriction fragment length polymorphism-polymerase chain reaction [RFLP-PCR], nox RFLP-PCR assays). RESULTS: Seventeen cases were diagnosed: seven patients were infected by B. aalborgi, one by B. pilosicoli, two by both species and four by Brachyspira spp. diagnosed both histopathology and microbiology (culture and molecular methods: 16S RFLP-PCR and nox RFLP-PCR assays). Three cases were referred to as Brachyspira spp. infections using only histopathology, including TEM. CONCLUSIONS: Our results demonstrated that intestinal spirochetosis, although rarely occurring, might play a role in chronic diarrhea and suggested a pathogenetic mechanism of intestinal spirochetosis based on the destruction of colonic microvilli and colitis histologically documented, providing additional clinical and pathological information on this entity. This study suggests that metronidazole seems to be the drug of choice for the eradication of intestinal spirochetosis. PMID- 17914950 TI - Age and distal colonic findings determine the yield of advanced proximal neoplasia in Chinese patients with rectal bleeding. AB - BACKGROUND AND AIMS: Few data were available on the optimal diagnostic strategy for Chinese patients with hematochezia. We aimed to evaluate the impact of age and distal colonic findings on the yield of diagnostic strategies in young Chinese patients with hematochezia. METHODS: Consecutive outpatients aged less than 50 years were analyzed using a hypothesized mixed diagnostic strategy to determine the optimal cut-off age for the use of sigmoidoscopy and colonoscopy. The efficacy and cost of the diagnostic strategy and the number of colonoscopies needed to detect one advanced proximal neoplasm (APN) using different cut-off ages were assessed. RESULTS: In the hypothesized mixed diagnostic strategy for young patients, the sensitivities for the detection of APN were 100%, 92% and 75% if the cut-off ages were 30, 35 and 40 years, respectively. The cost needed to detect one APN would be $US 3155, $US 3179 and $US 3497 if the cut-off ages were 30, 35 and 40 years, respectively. Colonoscopy would be performed in 84%, 69% and 51% of patients if the cut-off ages were 30, 35 and 40 years, respectively. CONCLUSION: Colonoscopy should be considered for Chinese patients with rectal bleeding who are aged > or =35 years or those aged <35 years who have adenoma in the distal colon. PMID- 17914951 TI - Comparison of malignant potential between serrated adenomas and traditional adenomas. AB - BACKGROUND: Serrated adenoma is a discrete colorectal epithelial neoplastic lesion that can evolve into colorectal cancer. However, the degree of malignant potential has not been firmly established as yet. The purpose of the present paper was to compare the malignant potential and clinicopathological features between serrated and traditional adenomas. METHODS: A total of 124 serrated adenomas from 116 patients were assessed, and 419 traditional adenomas from 200 were randomly selected. The combination of nuclear dysplasia and serration of > or =20% of crypts was regarded as serrated adenoma. The clinicopathological features of serrated and traditional adenomas were compared, and multivariate analysis performed to confirm whether the malignant potential of serrated adenoma was similar to that of traditional adenoma. RESULTS: The differences in age, sex, total number of adenomas, and synchronous lesions including adenoma with high grade dysplasia and carcinoma between subjects with and without serrated adenoma were not significant. Serrated adenomas were more frequently located in the rectum and sigmoid colon (P < 0.001), and the average size of serrated adenomas was greater than that of traditional adenomas (P < 0.05). The incidence of malignant lesions including high-grade dysplasia and carcinoma in serrated adenomas was found to be lower than in traditional adenomas (3.2% vs 9.3%, P < 0.05). In the multivariate analysis, adenoma type and polyp size constituted the risk factors for the incidence of high-grade dysplasia and carcinoma. CONCLUSIONS: Serrated adenoma is a premalignant lesion, but it has a lower potential for the development of malignancy than traditional adenomas. PMID- 17914952 TI - In vivo and in vitro antioxidant activity of ghrelin: Attenuation of gastric ischemic injury in the rat. AB - BACKGROUND AND AIM: Gherlin, an endogenous ligand for the growth hormone secretagogue receptor (GHS-R), is produced by stomach cells. It regulates food intake, gastric secretion and motility. However, its role as a protective agent in gastric ischemia/reperfusion (I/R) injury has not yet been investigated. Therefore, the objectives of the present study were to: (i) test the in vivo effect of peripherally administered ghrelin on gastric I/R-induced lesions in rats; and (ii) investigate in vitro the effect of ghrelin on reactive oxygen species (ROS) production by human polymorphoneuclear (PMN) cells. METHODS: The present study was carried out on three groups of rats (six per group): control (sham-operated), I/R (clamping of celiac artery for 30 min and reperfusion for 1 h), and I/R + ghrelin (200 ng/kg i.v., 15 min before ischemia and before reperfusion, respectively). Histological assessment of hematoxylin and eosin stained sections was performed and immunostaining with inducible nitric oxide (iNOS) antibody were performed on a gastric paraffin embedded section. Oxidative stress markers thiobarbituric acid reactive substance (TBARS) and glutathione (GSH) were measured in gastric tissue homogenates. Serum lactic acid dehydrogenase (LDH) was determined. Tumor necrosis factor-alpha (TNF-alpha) was assayed in gastric tissue homogenate. Gastric permeability was assessed calorimetrically using Evans blue dye. In vitro studies were carried out on isolated human PMN cells incubated with ghrelin and tested for ROS generation as measured by chemiluminecence (CL). RESULTS: Peripheral administration of ghrelin attenuated gastric injury by reducing ulceration, tissue congestion, cellular infiltration and vascular permeability. Serum level of LDH and tissue content of TNF-alpha were markedly reduced. A decrement in TBARS and an increment in GSH were observed. Ghrelin treatment attenuated iNOS protein expression which was upregulated by gastric ischemic injury. In vitro studies showed for the first time that ghrelin inhibited ROS generation by human PMN in a dose-dependent manner. CONCLUSIONS: These results provide evidence that peripherally administered ghrelin protects against gastric I/R injury. We also demonstrated that this protection is possibly accomplished through the antioxidant activity of ghrelin observed in vivo and in vitro. PMID- 17914953 TI - Effects of a novel histamine H2-receptor antagonist, lafutidine, on the mucus barrier of human gastric mucosa. AB - BACKGROUND AND AIM: Lafutidine is a novel histamine H(2)-receptor antagonist used primarily as an antisecretory agent in Japan. Previous human studies have not assessed its gastroprotective effects. The purpose of the present study was to determine the effects of lafutidine on the human gastric mucus layer using both histological and biochemical methods. METHODS: Of the 14 patients scheduled for gastrectomy who consented to participate, seven were given 14 days of lafutidine 20 mg/day (lafutidine group) and the others received no medication (control group). The surface mucus gel layer in Carnoy-fixed tissue sections was examined immunohistochemically. Both the thickness of the mucus layer and its mucin content were measured in gastric corpus mucosa. RESULTS: There was no detectable difference between the groups in the grade of gastritis or the immunohistochemical staining characteristics. The laminated structure of the surface mucus gel layer was retained after administration of lafutidine and it was thicker than the layer in the control group. The surface layer in the lafutidine group had three-fold more mucin than that in the control group. There was no difference between the two groups in the mucin content of the deep mucosa. CONCLUSION: Lafutidine, given at clinical dosages, not only inhibits acid secretion but also strengthens the mucus barrier of the human gastric mucosa. PMID- 17914954 TI - Rabeprazole delays gastric emptying of a nutrient liquid. AB - BACKGROUND: It is generally considered that gastric acid suppression delays gastric emptying of solid meals because gastric hypoacidity impairs peptic digestion and antral triturition. Rabeprazole is one of the most potent acid suppressants. We conducted this cross-over study to investigate if rabeprazole delays gastric emptying of liquid nutrients, for which peptic digestion is unnecessary. METHODS: On two randomized occasions, 13 healthy male volunteers underwent the (13)C-acetate breath test following ingestion of a 300 kcal-liquid meal. On one occasion, they had received 20 mg rabeprazole for the preceding 2 days and 1 h before the test. On another, they took no pretreatment. Based on 4 h breath samples, the half (13)CO(2) excretion time (t(1/2b)) and the time of maximal excretion (t(max)) were calculated as overall measures of gastric emptying. In addition, time profiles of gastric emptying were generated using the Wagner-Nelson analytical method, which creates the gastric emptying curve as accurately as the scintigraphy. RESULTS: Rabeprazole significantly prolonged t(max) but unchanged t(1/2b). The gastric emptying profiles showed that rabeprazole evoked a segmental slowing of gastric emptying during 0.5-1.25 h of post-meal ingestion. CONCLUSIONS: Rabeprazole suppresses gastric emptying of a liquid nutrient 0.5 h after meal ingestion. We have speculated that: (i) rabeprazole delays gastric emptying via a mechanism other than acid-pepsin maldigestion; and (ii) rabeprazole may intensify the post-gastric feedback regulation system (>0.5 h of post-meal). PMID- 17914955 TI - Modified allele-specific primer-polymerase chain reaction method for analysis of susceptibility of Helicobacter pylori strains to clarithromycin. AB - BACKGROUND AND AIM: Most clarithromycin-resistant strains of Helicobacter pylori have a mutation from adenine (A) to guanine (G) at position 2142 or 2143 of the 23S rRNA gene. Our aim in this study was to develop a polymerase chain reaction (PCR)-based assay that could determine these mutations in a single reaction tube. METHODS: We designed the forward primer FP2143G and the reverse primer RP2142G, which specifically anneal with the 2143G- and 2142G-mutated sequences, respectively, of the 23S rRNA gene of H. pylori. We also designed the forward primer FP-1 and reverse primer RP-1 upstream and downstream from the positions 2142 and 2143, respectively, to distinguish the wild-type A2142G and A2143G mutations from each other by amplicon sizes. DNA was extracted from 292 gastric tissue samples positive for rapid urease test, and the DNA underwent the PCR reaction. The results were compared with minimum inhibitory concentrations (MIC) for clarithromycin. RESULTS: Helicobacter pylori strains with A2142G, A2143G and wild type could be distinguished by amplicon sizes by a single PCR reaction. The genotyping results were correlated well with the MIC values for clarithromycin. The median MIC for clarithromycin of the wild-type strains was <0.015 microg/mL. Those of strains with 2142G or 2143G were > or =1.0 microg/mL. CONCLUSION: Our new PCR-based assay for 23S rRNA mutations of H. pylori is a useful method for detecting clarithromycin-resistant strains of H. pylori easily. PMID- 17914956 TI - Environmental factors of celiac disease: cytotoxicity of hulled wheat species Triticum monococcum, T. turgidum ssp. dicoccum and T. aestivum ssp. spelta. AB - BACKGROUND AND AIM: In the present paper, the toxicity of prolamines derived from three cereals with a different genome was investigated in human colon cancer Caco 2/TC7 and human myelogenous leukemia K562(S) cells. The purpose of this study was to investigate if species from ancient wheat could be considered as healthy food crops devoid or poor in cytotoxic prolamines for celiac disease. METHODS: Cytotoxicity was measured in terms of inhibition of cell growth, activation of apoptosis, release of nitric oxide (NO), detection of tissue transglutaminase (TG II) and alteration of transepithelial electrical resistance (TEER) on Caco-2/Tc7 and K562 (S) cell agglutination. Peptic-tryptic (PT) digest from bread wheat (T. aestivum S. Pastore) was used as a positive control. RESULTS: PT digests of prolamins from spelt wheat (T. aestivum ssp. spelta) were found to exert toxic effects on Caco-2/TC7 cells and to agglutinate K562(S) cells. Increased amounts of NO and TG II expression were observed in Caco-2/TC7 cells exposed to 1 mg/mL of spelt prolamins, suggesting that spelt wheat can induce cellular mechanisms implicated in the pathogenesis of celiac disease. By contrast, the PT digests from monoccum wheat (Triticum monococcum) and farro wheat (T. turgidum ssp. dicoccum) did not exhibit any negative effects on Caco-2/TC7 and K562(S) cells. CONCLUSIONS: The results have shown a constant and significant toxic effect of spelt wheat which is not shared by the two other ancient cereals. Future studies on celiac intestinal organ cultures are needed to increase the prospects of breeding programs aimed at developing wheat cultivars potentially tolerated by most celiac patients. PMID- 17914957 TI - Investigation into the signal transduction pathway via which heat stress impairs intestinal epithelial barrier function. AB - BACKGROUND AND AIMS: Intact protein absorption is thought to be a causative factor in several intestinal diseases, such as food allergy, celiac disease and inflammatory bowel disease. However, the mechanism remains unclear. The aim of this study was to characterize a novel signal transduction pathway via which heat stress compromises intestinal epithelial barrier function. METHODS: Heat stress was carried out by exposing confluent human intestinal epithelial cell line T84 cell monolayers to designated temperatures (37-43 degrees C) for 1 h. Transepithelial electric resistance (TER) and permeability to horseradish peroxidase (HRP, molecular weight = 44 000) were used as indicators to assess the intestinal epithelial barrier function. Phosphorylated myosin light chain (MLC), MLC kinase (MLCK) and protein kinase C (PKC) protein of the T84 cells were evaluated in order to identify the signal transduction pathway in the course of heat stress-induced intestinal epithelial barrier dysfunctions. RESULTS: The results showed that exposure to heat stress significantly increased intact protein transport across the intestinal epithelial monolayer; the amount of phospho-PKC, phospho-MLCK and phospho-MLC proteins in T84 cells decreased significantly at 41 degrees C and 43 degrees C although they increased at 39 degrees C. The heat stress-induced T84 monolayer barrier dysfunction was inhibited by pretreatment with PKC inhibitor, MLCK inhibitor, or HSP70. CONCLUSION: Heat stress can induce intestinal epithelial barrier dysfunction via the PKC and MLC signal transduction pathway. PMID- 17914959 TI - Effect of heme oxygenase-1 induction by octreotide on TNBS-induced colitis. AB - BACKGROUND AND AIM: Ulcerative colitis is a chronic inflammatory disease of the colon and rectum. Although the precise etiology of ulcerative colitis remains unknown, it is believed to involve an abnormal host response to endogenous or environmental antigens, genetic factors, and oxidative damage. The aim of the present study was to investigate whether heme oxygenase-1 (HO-1) induction by octreotide could protect against oxidative and inflammatory damage from induced colitis. METHODS: Rats received octreotide 50 microg/kg per day intraperitoneally for 5 days before 2,4,6 trinitrobenzene sulfonic acid (TNBS) solution administration and for 15 days following TNBS solution administration. Rats were killed on day 21, and colonic malondialdehyde (MDA) levels, glutathione (GSH) levels and HO-1 expression were measured. Nuclear factor (NF)-kappaB and HO-1 expression was evaluated by immunohistochemical examination of the colonic tissue. RESULTS: Rats with TNBS-induced colitis had significantly increased colonic MDA levels and HO-1 expression in comparison to the control group. Octreotide treatment was associated with increased HO-1 expression and GSH levels, but decreased MDA levels. Histopathological examination revealed that the intestinal mucosal structure was preserved in the octreotide-treated group. In addition, treatment with octreotide significantly increased HO-1 expression and decreased NF-kappaB expression by immunohistochemistry when compared to the TNBS induced colitis group. CONCLUSION: Octreotide appears to have protective effects against colonic damage in TNBS-induced colitis. This protective effect is, in part, mediated by modification of the inflammatory response and the induction of HO-1 expression. PMID- 17914958 TI - Butter feeding enhances TNF-alpha production from macrophages and lymphocyte adherence in murine small intestinal microvessels. AB - BACKGROUND AND AIM: Dietary fat is known to modulate immune functions. Intake of an animal fat-rich diet has been linked to increased risk of inflammation; however, little is known about how animal fat ingestion directly affects intestinal immune function. The objective of this study was to assess the effect of butter feeding on lymphocyte migration in intestinal mucosa and the changes in adhesion molecules and cytokines involved in this effect. METHODS: T-lymphocytes isolated from the spleen were fluorescence-labeled and injected into recipient mice. Butter was administered into the duodenum, and villus microvessels of the small intestinal mucosa were observed under an intravital microscope. mRNA expression of adhesion molecules and cytokines in the intestinal mucosa were determined by quantitative PCR. The effect of butter feeding on tumor necrosis factor (TNF)-alpha mRNA expression of intestinal macrophages was also determined. RESULTS: Intraluminal butter administration significantly increased lymphocyte adherence to intestinal microvessels accompanied by increases in expression levels of adhesion molecules ICAM-1, MAdCAM-1 and VCAM-1. This accumulation was significantly attenuated by anti-MAdCAM-1 and anti-ICAM-1 antibodies. Butter administration significantly increased TNF-alpha in the lamina proprial macrophages but not interleukin-6. Anti-TNF-alpha treatment attenuated the enhanced expression of adhesion molecules induced by butter administration. CONCLUSION: T-lymphocyte adherence to microvessels of the small intestinal mucosa was significantly enhanced after butter ingestion. This enhancement is due to increase in expression levels of adhesion molecules of the intestinal mucosa, which is mediated by TNF-alpha from macrophages in the intestinal lamina propria. PMID- 17914960 TI - Alpha-lipoic acid modulates gut inflammation induced by trinitrobenzene sulfonic acid in rats. AB - BACKGROUND AND AIM: Alpha-lipoic acid (ALA) has been shown to combat oxidative stress by quenching a variety of reactive oxygen species. It is involved in the regeneration of exogenous and endogenous antioxidants, chelation of metal ions, and repair of oxidized proteins. This study aimed to evaluate the potential beneficial effect of ALA on trinitrobenzenesulfonic acid (TNBS)-induced gut ileitis and colitis in rats. METHOD: After 48 h of fasting, Sprague-Dawley rats underwent a laparotomy under ether anesthesia. TNBS solution 30 mg/mL in 40% ethanol (1 mL) was injected into the lumen, 10 cm proximal to the ileocolonic junction to induce ileitis or intrarectally 8 cm proximal to the anal sphincter to induce colitis. ALA (25 mg/kg intraperitoneally, twice a day) was given after induction of inflammation and continued for 3 days. All animals were decapitated 3 days after induction of the inflammation. The mucosal lesions of the ileum and colon were scored macroscopically and microscopically. Samples were taken for the measurement of malondialdehyde (MDA) and glutathione (GSH) levels, tissue associated myeloperoxidase (MPO) activity and luminol- or lucigenin-enhanced chemiluminescence (CL). RESULTS: Macroscopic scores, morphological changes and increased tissue lipid peroxidation with a concomitant reduction in GSH of the ileitis or colitis groups were all reversed by treatment with ALA. ALA treatment was also effective in improving tissue MPO activity and CL values, which were elevated in untreated ileitis or colitis groups. CONCLUSION: ALA is beneficial in TNBS-induced gut inflammation in rats via suppression of neutrophil accumulation, preservation of endogenous glutathione and inhibition of reactive oxidant generation. PMID- 17914961 TI - Thymosin alpha 1 improves severe acute pancreatitis in rats via regulation of peripheral T cell number and cytokine serum level. AB - AIM: The aim of this study was to investigate the effect of thymosin alpha 1 (TA1) on severe acute pancreatitis (SAP) in rats. METHODS: Healthy Sprague-Dawley rats (n = 72) were randomly divided into four groups: control group, SAP group, and two TA1 treated groups. SAP was induced by injection of 5% sterile sodium taurocholate into the biliopancreatic duct (BPD), after which TA1 was given subcutaneously at 0 and 2 h at a dose of 100 microg/kg. The rats were killed at 3, 6 and 12 h, respectively. Serum amylase and lipase, interleukin (IL)-1beta, tumor necrosis factor-alpha (TNF-alpha), pancreatic wet/dry weight ratio and the percentage of CD3/CD4+/CD8+ T cells in peripheral blood mononuclear cells (PBMC) were measured. Next, 30 rats were randomly divided into three groups (each group containing 10 animals): SAP group (S) and two TA1 treated groups. The effects of TA1 on the survival of SAP were assessed 72 h after the induction of SAP. RESULTS: There was no significant change in the serum amylase and lipase levels after TA1 administration. Levels of serum IL-1beta, TNF-alpha and pancreatic wet/dry weight ratio were significantly reduced after TA1-treatment. Application of TA1 significantly balanced CD3/CD4+/CD8+ T cells of PBMC and improved histological scores and the survival rate. CONCLUSION: TA1 can reduce pancreatic inflammation by regulating differentiation of CD3/CD4+ T cells and decreasing the release of cytokines, thus attenuates pancreatic severity in SAP rats. PMID- 17914962 TI - Effects of octreotide in acute hemorrhagic necrotizing pancreatitis in rats. AB - BACKGROUND AND AIM: Octreotide is considered to reduce exocrine pancreatic secretion in acute hemorrhagic necrotizing pancreatitis decreasing pancreatic autodigestion. The aim of this study was to determine whether octreotide also has antioxidative effects in acute pancreatitis. Additionally time and dose of application were of interest. METHOD: Ninety male Sprague-Dawley rats were randomized into six groups (n = 15). Group 1 underwent a laparotomy, and animals in groups 2-6 received intraductal glycodeoxycholic acid followed by intravenous cerulein. Groups 3 and 4 were injected with 0.5 mg octreotide, while groups 5 and 6 received continuous intravenous infusion of 0.05 mg octreotide/h for 10 h. Treatment was initiated 6 hours after induction of pancreatitis (IP) in groups 3 and 5, and 14 h after IP in groups 4 and 6. At 24 h after IP all animals were killed and each pancreas was analyzed histopathologically. In addition, levels of pancreatic lipid peroxidation protective enzymes glutathione-peroxidase (GSH-Px) and superoxide dismutase (SOD) as well as lipid peroxidation via thiobarbituric acid reactive substances (TBARS) were determined. RESULTS: Early bolus application of octreotide reduced severity of histopathological changes in acute pancreatitis and decreased lipid peroxidation in pancreatic tissue samples; however, late bolus application and continuous intravenous infusion did not influence pancreatitis or lipid peroxidation. CONCLUSION: Octreotide seems to have a dose- and time-dependent effect on histopathology and lipid peroxidation in a model of pancreatitis in rats. PMID- 17914963 TI - Further studies on the arrest of cell proliferation in tumor cells at the invading front of colonic adenocarcinoma. AB - BACKGROUND AND AIM: The author has previously reported that neoplastic glands at the leading invading edge of colorectal carcinomas often display flat tumor cells and cellular gaps called glandular pores. The aim of this study was to audit cell proliferation and p53 mutation in flat tumor cells and in tumor cells at the tip of glandular pores at the leading invading front of colonic carcinomas. METHODS: Sections from 40 colonic adenocarcinomas were immunostained with the proliferation marker Ki67 and with p53 protein. Expression was assessed at the leading invading front in consecutive neoplastic glands having flat tumor cells and epithelial pores and in neoplastic glands showing neither flat tumor cells nor glandular pores. RESULTS: Flat tumor cells in neoplastic glands usually showed no Ki67 expression but overexpressed p53 mutation. In neoplastic glands with pores 40% of the tumor cells at the tip of the pores showed no Ki67, but they overexpressed p53 mutation. CONCLUSIONS: The results showed, for the first time, that p53-positive flat neoplastic colonic cells arrest their proliferation at the invading front. It is possible that these p53-positive/Ki67-negative neoplastic cells were temporarily removed from the cell cycle (G0). This paradoxical biological behavior of tumor cells might be connected with the formation of glandular pores and appears to indicate that arrest of cell proliferation at the advancing tumor front in colonic carcinomas occurs independently of p53 mutation. The possible existence of two independent molecular systems at the advancing tumor edge of colonic carcinomas, one supervising cell proliferation and the other zealously transferring the p53 mutation to daughter cells, is suggested. PMID- 17914964 TI - Transmission of hepatitis C virus by needle-stick injury in community settings. AB - BACKGROUND: Hepatitis C virus (HCV) is predominantly transmitted by blood-to blood contact, typically by sharing of needles by injecting drug users. Discarded needles could act as a vector for transmission of this infection. METHODS: Two cases of HCV seroconversion following a needle-stick injury in a community setting were identified. The effects of specimen processing and storage conditions on detection of HCV RNA were assessed to provide information about the likelihood of discarded needles containing infectious HCV. RESULTS: Consistent with a role for discarded needles in viral transmission, in vitro studies demonstrated that viral load declined by less than one log following storage for 24 h. CONCLUSION: All needle-stick injuries should be promptly investigated by serology and HCV-PCR. PMID- 17914965 TI - Hepatic iron accumulation is associated with disease progression and resistance to interferon/ribavirin combination therapy in chronic hepatitis C. AB - BACKGROUND AND AIMS: Liver iron accumulation in patients with chronic hepatitis C (CHC) has received increasing attention in recent years. The aim of this study was to determine the prevalence and severity of liver iron deposition in CHC, to assess its relationship with clinical, biochemical and histological characteristics, and to study its influence on the response to interferon (IFN) plus ribavirin combination therapy. METHODS: We studied liver biopsy specimens from 103 hepatitis C virus (HCV) and 34 hepatitis B virus (HBV) infected patients and total iron score (TIS) was measured. Seventy patients infected with HCV genotype 1b were treated with IFN/ribavirin for 24 weeks. RESULTS: CHC patients had a significantly higher TIS than chronic hepatitis B (CHB) patients (7.03 +/- 5.34 vs 4.41 +/- 4.49, P = 0.0056). TIS was significantly correlated with alcohol intake (P = 0.0213, r = 0.290), transaminase level (P = 0.0126, r = 0.247), platelet count (P = 0.0002, r = -0.369), histological grading (P = 0.0121, r = 0.248) and staging (P = 0.0003, r = 0.356) in CHC patients. Pretreatment TIS was significantly higher in non-sustained virological responders (SVR) than in SVR to IFN/ribavirin treatment (TIS = 7.69 +/- 5.76 vs 4.39 +/- 3.27, P = 0.0310). Multiple regression analysis showed that TIS was the only independent variable associated with resistance to IFN/ribavirin (P = 0.0277). CONCLUSIONS: Liver iron deposition was common in CHC compared to CHB and was associated with liver disease progression. Increased hepatic iron stores in CHC were related to resistance to IFN/ribavirin treatment. PMID- 17914966 TI - Effect of lactoferrin in patients with chronic hepatitis C: combination therapy with interferon and ribavirin. AB - OBJECTIVES: Lactoferrin has been reported to inhibit hepatitis C virus (HCV) infection in cultured human hepatocytes and HCV viremia in patients with chronic hepatitis C (CHC). The aim of this study was to evaluate the effect of combined triple therapy of lactoferrin, interferon and ribavirin in patients with CHC. METHODS: A total of 111 Japanese patients with CHC were randomly assigned to a lactoferrin group (n = 50) and a control group (n = 61). The lactoferrin group was treated with lactoferrin for 8 weeks and then with lactoferrin, interferon and ribavirin for 24 weeks; the control group was treated with interferon and ribavirin for 24 weeks. Serum anti-lactoferrin antibody, clinical and laboratory measurement were determined. RESULTS: The mean HCV RNA titer significantly decreased at the end of lactoferrin monotherapy. Sustained virological response to therapy was significantly higher (P < 0.05) in the lactoferrin responder group (55%) than in the control group (18%). CONCLUSIONS: The results show that the decrease in HCV RNA titer by lactoferrin monotherapy contributes to the effectiveness of the combined therapy of interferon and ribavirin in patients with CHC. Lactoferrin is a potential useful adjunct treatment for patients with CHC. PMID- 17914967 TI - Mutations in ISDR of NS5A gene influence interferon efficacy in Chinese patients with chronic hepatitis C virus genotype 1b infection. AB - BACKGROUND AND AIM: The interferon sensitivity determining region (ISDR) in the non-structural 5A protein (NS5A) gene of hepatitis C virus genotype 1b (HCV-1b) has been demonstrated in Japanese patients; however, contradictory data have been reported from other parts of the world, particularly from Europe and the USA. This study investigated whether mutations in the ISDR of NS5A gene influence interferon efficacy in Chinese patients with chronic HCV-1b infection. METHODS: Forty-five Chinese patients with chronic HCV infection were enrolled in a retrospective study. Of these patients, 20 with HCV-1b infection completed an interferon monotherapy course and 6-month follow-up. The pretreatment mutations in the nucleic acid sequence of NS5A 2209-2248 (ISDR) of HCV-1b were identified by sequencing RT-PCR products followed by comparison with the prototype sequence (HCV-J). Correlation with the clinical efficacy of interferon alpha therapy was then analyzed retrospectively. RESULTS: The statistical analysis indicated a significant correlation between the mutations in ISDR of HCV-1b and sustained virological response to interferon alpha treatment (P < 0.05), and an association of the mutations in ISDR with the pretreatment serum alanine aminotransferase levels (P < 0.01) in Chinese patients with chronic HCV-1b infection. CONCLUSIONS: The results support the concept that the response of HCV-1b infected patients towards interferon therapy is influenced by the mutations within the ISDR of NS5A gene. PMID- 17914968 TI - Role of AST to platelet ratio index in the detection of liver fibrosis in patients with recurrent hepatitis C after liver transplantation. AB - BACKGROUND AND AIM: Per protocol annual liver biopsy represents the gold standard in the assessment of graft fibrosis progression due to recurrent hepatitis C after liver transplantation. Non-invasive liver fibrosis tests have been proposed as surrogate markers of liver fibrosis in hepatitis C virus (HCV)-positive immune competent patients. No data are available in the literature on the usefulness of non-invasive liver fibrosis tests in liver transplanted patients with recurrent HCV infection. METHODS: A total of 102 annual per protocol liver biopsies performed in 51 consecutive HCV-positive recipients (31 men), with a follow-up period lasting up to 5 years, were included and evaluated in this study. At each time point, the following non-invasive liver fibrosis tests were calculated: aspartate aminotransferase (AST) to alanine aminotransferase (ALT) ratio, age platelet index, AST to platelet ratio index (APRI), Forns' fibrosis index and Bonacini's discriminant score. RESULTS: In discriminating patients with histological fibrosis score >2, APRI provided the best area under the receiver operating characteristic curves (AUROC) (0.801), in comparison to the other four non-invasive liver fibrosis tests. The AUROC of APRI was better in female (0.871) than in male (0.753) recipients. Among female recipients, an APRI value >1.4 was 91% sensitive and 75% specific in detecting a staging score >2. The corresponding values among male recipients were 60% and 77%, respectively. CONCLUSIONS: Among non-invasive liver fibrosis tests, APRI has the highest diagnostic value in discriminating liver transplanted patients with progression to significant liver fibrosis, although its accuracy is influenced by recipient sex. PMID- 17914969 TI - Prediction of large esophageal varices in patients with cirrhosis of the liver using clinical, laboratory and imaging parameters. AB - BACKGROUND: It is currently recommended that all patients with liver cirrhosis undergo upper gastrointestinal endoscopy (UGIE) to identify those who have large esophageal varices (LEVx) that carry a high risk of bleeding and may benefit from prophylactic measures. This approach leads to unnecessary UGIE in those without LEVx. We tried to identify clinical, laboratory and imaging parameters that may predict the presence of LEVx and help select patients for UGIE. METHODS: This prospective study included newly diagnosed patients with cirrhosis and no history of gastrointestinal bleeding scheduled to undergo UGIE. Patients underwent detailed clinical examination, blood tests (hematology, liver function tests) and ultrasonography. Size of esophageal varices was assessed at UGIE; Paquet's grades 0-II were classified as small varices, and III-IV as LEVx. Association of LEVx with qualitative and quantitative parameters was studied using chi(2) and Mann Whitney U-tests, respectively. Parameters found to be significant were tested in a forward-conditional multivariate logistic regression analysis to identify independent predictors. Receiver operating characteristic curve analysis was used to assess the efficacy of prediction models. RESULTS: Of the 101 patients (median age 45; range 15-74 years; 87 male; Child-Pugh class: A 18, B 31, C 52), 46 had LEVx. On univariate analysis, five variables were significantly associated with the presence of LEVx. These included pallor (P = 0.026), palpable spleen (P = 0.009), platelet count (P < 0.002), total leukocyte count (P < 0.0004) and liver span on ultrasound (P = 0.031). On multivariate analysis, two of these parameters, namely low platelet count and presence of palpable spleen, were found to be independent predictors of the presence of LEVx. A receiver-operating characteristics curve using the predictor function arrived at from this analysis had an area under the curve of 0.760. CONCLUSION: Presence of palpable spleen and low platelet count are independent predictors of presence of LEVx in patients with cirrhosis. Use of these parameters may help identify patients with a low probability of LEVx who may not need UGIE. This may help reduce costs and discomfort for these patients and the burden on endoscopy units. PMID- 17914970 TI - Involvement of RhoA in progression of human hepatocellular carcinoma. AB - BACKGROUND AND AIM: The activation of Rho proteins has been shown to lead to loss of polarity in cancer cells, as well as reorganization of the cytoskeleton and facilitation of cell motility, possibly resulting in their malignant potential. The clinicopathological significance of RhoA, however, is not yet well known in the case of hepatocellular carcinoma (HCC). This study evaluated the clinicopathological correlation of RhoA levels with HCC. METHODS: The intratumor expression level of Rho was determined and compared with that in adjacent non cancerous hepatic tissue using quantitative real time RT-PCR and Western blotting in 64 patients with HCC. Relationships between the level of RhoA and clinicopathological factors were examined. RhoA protein expression was confirmed by immunohistochemistry. RESULTS: RhoA immunostaining was strong in malignant tissue whereas it was minimal in benign tissue. Tumor tissue of HCC patients demonstrated a copy number of RhoA mRNA that was well correlated with its protein level in each case and was significantly higher than that found in the corresponding non-cancerous liver tissue (P < 0.01). With regard to venous invasion, satellite lesions and advanced pTNM stage, the RhoA level tended to be higher in HCC than that seen in negative tissue (P < 0.05). CONCLUSIONS: This is the first demonstration that the expression level of RhoA is correlated with tumor progression and metastasis in HCC. RhoA in tumor tissue might be expected to be not only a good candidate marker for invasive and proliferative tumor cells, but also a molecular target of these cells. PMID- 17914971 TI - High prevalence of hepatitis B virus infection and inferior vena cava obstruction among patients with liver cirrhosis or hepatocellular carcinoma in Nepal. AB - BACKGROUND: Little is known about the prevalence of hepatitis B virus (HBV) DNA and the genotype distribution among patients with liver diseases in Nepal, where obstruction of the hepatic portion of the inferior vena cava (IVCO) is common. The aim of the present paper was to assess the roles of HBV infection and IVCO in liver cirrhosis (LC) and hepatocellular carcinoma (HCC) in Nepal. METHODS: Serum samples from 121 patients (89 male, 32 female; age, 55.0 +/- 13.6 years) with or without IVCO consisting of 70 LC patients and 51 HCC patients in Nepal, were tested for HBV-DNA. RESULTS: The HBV-DNA was detected in 68 patients (56%) including 20 hepatitis B surface antigen (HBsAg)-negative patients: 33 LC patients (47%) and 35 HCC patients (69%) had detectable HBV-DNA (P = 0.0303). Among the 89 patients with IVCO, HBV-DNA was detected in HCC patients significantly more frequently than in LC patients (80%vs 43%, P = 0.0005). The frequency of HBV viremia was significantly higher among HCC patients with IVCO than those without (80%vs 44%, P = 0.0236), and that of HBV viremia with IVCO was significantly higher among HCC patients than among LC patients (55%vs 27%, P = 0.0153). The HBV genotypes A and D were predominant, and genotype A was significantly more frequent among HCC patients than among LC patients (22%vs 6%, P = 0.0090). Among HCC patients, those with genotype A HBV were significantly younger than those with genotype D (43 +/- 13 vs 57 +/- 12 years, P = 0.0252). CONCLUSION: Hepatitis B virus alone (especially genotype A) or in concert with IVCO may be responsible for development of HCC in Nepal. PMID- 17914972 TI - Efficacy of antiviral therapy with lamivudine after initial treatment for hepatitis B virus-related hepatocellular carcinoma. AB - AIM: The aim of this study was to determine whether antiviral therapy with lamivudine is beneficial in patients after initial treatment for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). METHODS: Forty-nine consecutive patients with HBV-related HCC completely treated by hepatic resection or radiofrequency ablation were retrospectively enrolled in this study. Comparison was made between 16 patients who received lamivudine therapy at a dose of 100 mg/day after treatment for HCC (lamivudine group) and 33 patients who did not (control group) in terms of changes in remnant liver function, HCC recurrence and survival. RESULTS: Cumulative recurrence rates of HCC did not significantly differ between the two groups (P = 0.622). However, median Child-Pugh score at the time of HCC recurrence was significantly different; 5 (range 5-6) in the lamivudine group versus 7 (range 5-12) in the control group (P = 0.005). All patients in the lamivudine group were able to receive curative treatment for recurrent HCC. In contrast, 10 of 15 patients in the control group were unable to receive curative optimal therapy for recurrent HCC due to deterioration of remnant liver function. The cumulative survival rates of patients in the lamivudine group tended to be higher than those of patients in the control group (P = 0.063). CONCLUSION: It is suggested that lamivudine therapy is beneficial for patients after initial treatment for HBV-related HCC because it contributes to improving remnant liver function, thus decreasing the risk of liver failure and increasing the chances of receiving available treatment modalities for recurrent HCC. PMID- 17914973 TI - Comparison of surgical outcomes for small hepatocellular carcinoma in patients with hepatitis B versus hepatitis C: a Chinese experience. AB - BACKGROUND: Although both hepatitis B virus (HBV) and hepatitis C virus (HCV) infection are well recognized risk factors for hepatocellular carcinoma (HCC), little is known with respect to how HBV and HCV infection affect HCC recurrence in postoperative HCC Chinese patients. The objective of this study was to determine if differences exist in preoperative characteristics and postoperative HCC recurrence in patients with different HBV and HCV infection status. METHODS: The study population consisted of 413 patients undergoing a curative resection at Tianjin Cancer Hospital for small HCC (< or =3 cm) from January 1997 to December 2003. The patients were divided into four groups: HCV only (n = 75), HBV only (n = 251), HBV and HCV (n = 33), and neither HBV nor HCV (NBNC, n = 54). The preoperative status and postoperative HCC recurrence were recorded. Survival analyses were used to assess the impact of HBV/HCV status on HCC recurrence. RESULTS: Patients with HCV had a significant association with older age, lower mean preoperative platelet counts and albumin levels, higher mean prothrombin time, alanine aminotransferase and total bilirubin levels and multinodular tumors during diagnosis. Patients with HCV also had significantly less differentiated tumors and a higher incidence of vascular invasion and cirrhosis when compared to the other groups. During the follow-up, the HCV group showed a higher incidence of intrahepatic recurrence and multiple recurrent lesions than the other patients. CONCLUSIONS: Patients with HCV infection tended to be older, and were characterized by more severe cirrhosis and higher incidence of tumor multicentricity. The statistically significant determinants for reoccurrence in patients with small HCC were HCV infection, presence of vascular invasion and multiple tumors. PMID- 17914974 TI - Perioperative infection control and its effectiveness in hepatectomy patients. AB - BACKGROUND AND AIM: Post-hepatectomy infections require careful attention, because they may deteriorate into liver failure. We retrospectively reviewed such infections in cases without biliary or intestinal reconstruction. METHODS: This retrospective study involved 535 patients with liver tumors who underwent hepatectomy at the Department of Gastroenterological Surgery of Yokohama City University Hospital between April 1992 and March 2005. After classification into four groups depending on changes in infection countermeasures used during different periods, the treatment outcomes were examined. No such anti-infection measures were taken during the first period; a closed suction drainage system and early enteral nutrition after surgery were introduced between the first and second periods; thorough management of bile leakage and prevention of nosocomial infection were added between the second and third periods; and surgical site infection (SSI) surveillance together with absorbable sutures instead of silk sutures between the third and fourth periods. RESULTS: The incidence of postoperative infection decreased significantly with additional countermeasures: first period 44.7%; second period 24.1%; third period 15.0%; and fourth period 9.2%. The incidence of both SSI and remote infection were similarly reduced. Postoperative infection risk factors were age, presence of diabetes mellitus, the use of silk sutures and bile leakage, while those for SSI were the use of silk sutures and bile leakage. CONCLUSION: The incidence of postoperative infection and SSI were significantly reduced by our infection countermeasures, especially by bile leakage management and the use of absorbable sutures. PMID- 17914975 TI - Biliary tract malignancy and abnormal pancreaticobiliary junction in a Western population. AB - BACKGROUND AND AIM: Abnormal pancreaticobiliary junction is a rare congenital disorder in which the junction of biliary and pancreatic duct is located outside the duodenal wall and forms a long common channel. The disorder has been associated with a high incidence of gallbladder cancer in Japanese studies. The aim of this study was to assess the frequency of abnormal pancreaticobiliary junction in a Western population and its association with biliary tract malignancies. METHODS: All endoscopic retrograde cholangiopancreatography examinations of 82 consecutive patients with gallbladder carcinoma or biliary duct cancer treated at Baylor University during a period of 10 years were analyzed. A further 220 consecutive patients with normal films or non-malignant pancreaticobiliary disease acted as controls. RESULTS: Fifty-eight (70.7%) patients were found to have adequate imaging of junction and were included in the analysis. These included 37 patients with common bile duct carcinoma and 21 with gallbladder carcinoma. Abnormal pancreaticobiliary junction (common channel > or =8 mm) was observed in 44.8% patients with biliary tract carcinoma compared to 6.2% of controls (P < 0.01). Eighteen had type I abnormality and eight type II. Seven patients had a cystic dilatation of the common bile duct, all with type I abnormality. Mean overall survival was 9.5 months. Survival did not differ significantly between patients with normal and abnormal junctions (P = 0.1). CONCLUSIONS: The results suggest a close association between the anatomy of the distal ends of the common bile duct and main pancreatic duct and development of biliary tract carcinoma in Western populations. PMID- 17914976 TI - Management of liver hemangiomas according to size and symptoms. AB - BACKGROUND AND AIM: Liver hemangiomas are the most common benign liver tumors. These lesions are usually incidental findings during imaging studies of the abdomen performed for other reasons. The indication for surgical resection of these lesions remains controversial. METHODS: Records of patients referred for evaluation of radiologically and/or histopathologically proven liver hemangiomas, from June 1991 to February 2006, were retrospectively analyzed. Reason for referral, results of imaging studies, and surgical treatment and outcome were reviewed. RESULTS: There were 34 patients identified. The hemangioma size was <5 cm in 15 patients (44%) and >5 cm in 19 patients. The most common reason for referral was right upper abdominal pain in 59% (20/34) of patients. Abdominal ultrasound was conclusive in 66.7% (16/24) and four-phase computed tomography (CT) in 82.6% (19/23) of patients. Surgical resection was undertaken in 14 patients (41%) after a mean follow-up time of 36.5 months. The indication for treatment was progressive abdominal pain in 78.6% (11/14). Mean size of resected lesions was larger compared to non-resected lesions (10.3 vs 4.8 cm; P = 0.004). Postoperative morbidity occurred in three patients (21.4%). One patient had persisting abdominal pain after resection of an 8-cm hemangioma. Twenty patients were observed and showed no complications related to the liver hemangioma during follow-up. CONCLUSIONS: Liver hemangiomas can be readily diagnosed by ultrasound or multiphase contrast-enhanced helical CT. The indications for surgical resection are progressive abdominal pain in combination with size >5 cm. Observation is justified in patients with minimal or no symptoms, even in patients with giant hemangiomas. PMID- 17914977 TI - Hepatocyte dynamics in a three-dimensional rotating bioreactor. AB - BACKGROUND AND AIMS: The use of an artificial liver system with extracorporeal circulation or a three-dimensional bioreactor perfused with liquid culture medium inevitably exposes hepatocytes to fluid mechanical stress (MS). The expression of liver-specific hepatocyte functions seems to be modulated by the magnitude of MS. Nonetheless, few studies have focused on the direct effects of MS on hepatocytes. We subjected hepatocytes to MS using an MS loading device and investigated the effects on the cytoskeleton and hepatocyte dynamics inside three-dimensional scaffolds by monitoring the changes in actin fiber, one of the components of the cytoskeleton. We also assessed the influence of MS on specific hepatocyte functions. METHODS: We subjected hepatocytes to MS by a rotating radial flow bioreactor (RRFB) and examined the effects by comparing the MS-loaded culture cells with cells cultured under stationary conditions without MS loading. The hepatocytes (1 x 10(6)/cm(3)) were seeded on gauze without collagen coating and examined to determine morphological changes after 60 h incubation. Actin filaments in samples from the MS-loaded hepatocyte culture were stained by fluorescein isothiocyanate-labeled phalloidin. RESULTS: Hepatocyte aggregation was observed in the MS-loaded culture, but not in the unloaded stationary culture. Better albumin products were observed in the MS-loaded group than in the stationary culture group at all measurement points. Actin filaments extended toward the scaffold after the start of MS loading incubation and polymerized around the hepatocytes. The hepatocyte aggregation eventually advanced to the formation of spheroids. CONCLUSION: These results suggest that MS-induced polymerization of actin filaments stimulate hepatocyte aggregation and thereby improve hepatocyte-specific function. PMID- 17914978 TI - Protective effect of n-3 polyunsaturated fatty acid on primary culture of rat hepatocytes. AB - BACKGROUND AND AIM: Recently, we reported on the beneficial clinical effects of eicosapentaenoic acid (EPA) in patients with primary biliary cirrhosis (PBC) who were unresponsive to ursodeoxycholic acid (UDCA). In this study we examined the effect of EPA on rat hepatocytes in primary culture. METHODS: Hepatocytes were isolated from rat liver by perfusion of collagenase and cultured with or without EPA. Cell damage induced by chenodeoxycholic acid (CDCA) was assessed by WST-8 assay and lactate dehydrogenase (LDH) release. PGE(2) and LTB(4) concentrations in the culture medium were measured by enzyme-linked immunosorbent assay (ELISA). cDNA was made from total RNA that was extracted from hepatocytes, and TaqMan polymerase chain reaction (PCR) was performed to assess the expression of CuZn and Mn superoxide dismutase (SOD) mRNA. RESULTS: When rat hepatocytes were cultured in the presence of EPA, the damage caused by CDCA was significantly decreased compared with cells cultured without EPA. Cytotoxicity significantly decreased in the presence of EPA. Furthermore, SOD mRNA expression was increased by adding EPA. These findings indicated that EPA protects cells by scavenging superoxide radicals ((*)O(2-)) mediated by SOD production. CONCLUSION: EPA has a direct protective effect on rat hepatocytes, which is in agreement with the clinical efficacy of EPA in PBC patients. PMID- 17914979 TI - Ischemic preconditioning enhances regenerative capacity of hepatocytes in long term ischemically damaged rat livers. AB - BACKGROUND AND AIMS: Ischemic preconditioning (IPC) protects tissues against ischemia and reperfusion (I/R) injury. The aim of this study was to examine the impact of IPC on protection and regeneration of hepatocytes after prolonged I/R injury. METHODS: A rat model of segmental (70%) hepatic ischemia was used to determine the effect of 10-min IPC preceding 40, 60, 90, or 120 min of liver ischemia. The effect was assessed by comparing cytolysis markers and necrotic areas of the liver, as well as the regenerative capacity of hepatocytes using the proliferating cell nuclear antigen labeling index (PCNA-LI) and weight of the ischemic liver lobe. Protein kinase B/Akt (Akt) and caspase-9 were investigated immunohistochemically to determine the effect of IPC on activation of survival and anti-apoptotic signals. RESULTS: In the model of 40 min I/R, which resulted in focal necrosis of the liver, IPC significantly protected against I/R injury by reducing the area of focal necrosis, level of PCNA-LI and immunoreactivities to Akt and caspase-9. In contrast, IPC did not prevent ischemic damage in the 90- and 120-min ischemic model with massive liver necrosis. However, IPC enhanced the regenerative capacity of the remaining hepatocytes with higher levels of PCNA-LI, number of Akt-positive cells and mean weight of the liver lobe postoperatively than in the controls. CONCLUSIONS: In a model of focal necrosis of the liver, IPC protected hepatocytes against I/R injury. In addition, in a model of massive necrosis, IPC maintained the regenerative capacity of the remaining hepatocytes by enhancing the survival signals. PMID- 17914980 TI - Regulatory effects of corticosterone on ornithine decarboxylase activity during liver regeneration in rats. AB - AIMS: The regulation of ornithine decarboxylase (ODC) gene expression and enzyme activity by corticosterone during rat liver regeneration induced by partial hepatectomy (PH) was evaluated. METHODS: Bilateral adrenalectomies were performed on ether-anesthetized rats 3 days before PH. Corticosterone in sesame oil was injected subcutaneously to adrenalectomized rats. ODC mRNA, ODC protein and enzyme activity were detected by in situ hybridization, Western blot and high performance liquid chromatography (HPLC), respectively. RESULTS: The ODC mRNA levels, protein accumulation and enzyme activity were lower in the intact liver compared to the regenerating liver. After PH, mRNA levels were remarkably enhanced in all groups and peaked at 5 h post-PH, and presented a persistent increase only in adrenalectomy rats during the regeneration process. Corticosterone treatment resulted in a dose-dependent decrease in ODC mRNA content after 5 h post-PH. ODC protein accumulation in adrenalectomy rats was higher than that in sham-adrenalectomy rats, but it decreased in corticosterone treated (10 mg/kg) rats until 24 h post-PH, with a strong decline seen in 40 mg/kg corticosterone-treated rats. ODC activity was rapidly promoted, and the highest levels were observed at 6 h after PH in all groups. After corticosterone treatment, the activities declined significantly at 6 h post-PH, with the lowest value found in the 40 mg/kg group. CONCLUSIONS: Corticosterone treatment results in dose-dependent decreases in ODC mRNA and enzyme protein both in the intact liver and the regenerating liver. The change in ODC activity is partially related to alterations of ODC mRNA and protein accumulation. PMID- 17914981 TI - Initial site of bile regurgitation following extrahepatic biliary obstruction in living rats. AB - BACKGROUND AND AIM: The precise mechanism of bile regurgitation from the biliary system to the blood stream still remains to be elucidated. The aim of this study was to examine the initial site of bile regurgitation in vivo after common bile duct (CBD) obstruction by digitally enhanced fluorescence microscopy. METHODS: The fluorescence excreted into bile canaliculi after the administration of sodium fluorescein was continuously observed in CBD obstruction, using video-enhanced contrast (VEC) microscopy equipped with a silicon intensified target (SIT) camera. The liver histology and the localization of Mg(2+)-ATPase were examined by light and electron microscopy. RESULTS: By the continuous recording of canalicular fluorescence, the sequential regurgitation of the fluorescence from the canaliculi to the hepatocyte cytoplasm to the sinusoids was distinctively recognized after CBD obstruction. Bile canalicular fluorescence was enhanced, and then the fluorescence of the hepatocyte cytoplasm increased in intensity, followed by regurgitation of the fluorescence to the sinusoids. These in vivo sequences closely correlated with changes in CBD pressure. In zone 1, canalicular fluorescence focally burst into hepatocyte cytoplasm, thus resulting in the formation of fluorescent cells. By light and electron microscopy, the fluorescent cells were found to correspond to the liver cell injury. The reaction products of Mg(2+)-ATPase were incorporated into vesicles with a decreased canalicular activity, and then were transported to the sinusoidal surface after CBD obstruction. CONCLUSIONS: The initial site of bile regurgitation may be transcellular, and partly involves liver cell injury in zone 1 in extrahepatic biliary obstruction, associated with increased pressure of the biliary system. PMID- 17914982 TI - Gene expression profiling in whole liver of bile duct ligated rats: VEGF-A expression is up-regulated in hepatocytes adjacent to the portal tracts. AB - BACKGROUND AND AIM: It would be of clinical importance to clarify molecular mechanisms of cholangiocytes proliferation for the treatment of intractable cholestatic diseases. The aim of this study was to elucidate gene expression profiling in the whole liver of bile duct ligated (BDL) rats using microarray analysis. In addition, the localization and time course of up-regulated expression of vascular endothelial growth factor (VEGF) was investigated. METHODS: Male Sprague-Dawley rats were used. The whole liver was removed from BDL and sham-operated rats at day 2 after the procedure, and microarray analysis was performed using an array on which 3757 rat cDNA clones spotted. The up-regulation of VEGF expression was investigated by RT-PCR using livers at day 1, 2, 4 and 7, and immunoblotting and immunohistochemistry at day 2. RESULTS: Marked proliferation of bile ducts was observed in livers of BDL rats. By microarray analysis, 38 up-regulated and 17 down-regulated transcripts were detected in whole liver of the BDL rat. The expression of VEGF-A was significantly elevated in the BDL rats at day 2; the VEGF-A/GAPDH ratio was 4.030 +/- 2.493 in BDL rats and 1.159 +/- 0.125 in sham-operated rats (P = 0.0330). The up-regulation of VEGF A expression was maximal at day 2. Immunoblotting also demonstrated up-regulated expression of VEGF-A at the protein level. Immunostaining of VEGF revealed that the expression was evident in hepatocytes adjacent to the portal tracts, and scarcely observed in hepatocytes at the centrilobular area or cholangiocytes. CONCLUSION: Gene expression profiling in the whole liver of the BDL rats revealed 38 up-regulated and 17 down-regulated transcripts. In addition, the up-regulated expression of VEGF was mainly observed in hepatocytes surrounding to the portal tracts. PMID- 17914983 TI - Participation of bone marrow cells in biliary fibrosis after bile duct ligation. AB - BACKGROUND AND AIM: Bone marrow derived cells are involved in the process of hepatic fibrosis secondary to chronic injury. However, it is not yet known how quickly this event occurs in acute fibrosis models. The purpose of this study was to determine the role of bone marrow cells in rapid fibrosis following bile duct ligation in mice using green fluorescent protein (GFP) expressing bone marrow cells. METHOD: After whole body irradiation, 1 x 10(6) donor whole bone marrow cells from green fluorescent protein(+/-) mice were transplanted into C57BL/6 recipients via the tail vein. Four weeks after bone marrow transplantation, chimeric mice were subjected to common bile duct ligation, and livers of these animals were histologically examined after bile duct ligation using anti fibroblast specific protein (FSP)-1 antibody and anti-alpha-smooth muscle actin (alpha-SMA) antibody. RESULTS: Periductal fibrosis consisting of fibroblast specific protein-positive cells was demonstrated histologically as early as day 7. Most of the fibrotic cells were green fluorescent protein-negative, however, a significant number of cells were green fluorescent protein-positive and some were also anti-FSP or alpha-SMA-positive. CONCLUSION: Differentiation of bone marrow derived cells into activated fibroblast and myofibroblast-like phenotypes occurs in the very early course of periductal fibrosis following bile duct ligation, suggesting a new strategy for prevention of biliary fibrosis by inhibiting migration of bone marrow cells to liver. PMID- 17914984 TI - Preventive role of genistein in an experimental non-alcoholic steatohepatitis model. AB - BACKGROUND AND AIM: The aim of the present study was to evaluate the preventive role of genistein, a phytoestrogen with a wide variety of pharmacological effects, in an experimental non-alcoholic steatohepatitis (NASH) model. METHODS: Thirty-six Sprague-Dawley rats were divided into three groups. Group 1 (control) received only a standard rat diet, group 2 (placebo) was given a high fat diet (HFD) plus 0.5 mL/day saline subcutaneously, and group 3 (genistein group) a HFD plus subcutaneous genistein injection at dose of 0.2 mg/kg/day for 6 weeks. All rats were killed after 6 weeks. Serum aminotransferases, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta, and plasma and liver malondialdehyde (MDA) levels were measured. Additionally, steatosis, ballooning degeneration and inflammation of the liver were examined histopathologically. RESULTS: Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) (P < 0.001 for each), plasma and liver tissue MDA and plasma TNF-alpha levels (P < 0.001, <0.001, <0.01, respectively) were found to be higher in the placebo group than in the control group. TGF-beta levels, however, were comparable in the placebo and control groups (P > 0.05). Histopathologically, steatosis, inflammatory cells per mm(2) and ballooning degeneration were significantly higher in the placebo group than in the control group (P < 0.001 for each). Nevertheless, AST and ALT (P < 0.05 for each), plasma and liver tissue MDA (P < 0.05 for each) and plasma TNF-alpha levels (P < 0.001) were significantly decreased in the genistein group compared to the placebo group. Histopathologically, steatosis (P < 0.05), inflammatory cells per mm(2) and ballooning degeneration (P < 0.01 for each) in the genistein group were also significantly lower than in the placebo group. CONCLUSIONS: Genistein, a strong antioxidant agent, significantly decreased the plasma TNF-alpha level and remarkably prevented the emergence of NASH by improving the biochemical and histopathological abnormalities via attenuating oxidative stress. PMID- 17914985 TI - Rho/Rho kinase is a key enzyme system involved in the angiotensin II signaling pathway of liver fibrosis and steatosis. AB - BACKGROUND AND AIM: The molecular mechanisms underlying the involvement of the renin-angiotensin system in hepatic fibrosis are unclear. Recently, it was reported that a Rho kinase inhibitor prevented fibrosis of various tissues and that the Rho/Rho kinase pathway was involved in the renin-angiotensin system of vascular smooth muscle cells. In this study, the involvement of the Rho/Rho kinase pathway on angiotensin II signaling in liver fibrogenesis and generation of steatosis was investigated. METHODS: Rats were fed a choline-deficient/L-amino acid-defined (CDAA) diet continuously and treated with a Rho kinase inhibitor, Y 27632, and an angiotensin II receptor blocker, TCV-116. Liver histology and hepatic stellate cell activation were analyzed. Free radical production was detected by 4-hydroxynonenal and 8-hydroxy-2'-deoxyguanosine immunostaining and the expression of tumor necrosis factor-alpha was examined. Isolated hepatic stellate cells were pretreated with a Rho kinase inhibitor, Y-27632, or an angiotensin II receptor blocker, CV-11974, and stimulated with angiotensin II, and mRNA expression of transforming growth factor-beta and alpha-smooth muscle actin was analyzed. RESULTS: Both the angiotensin II receptor blocker and the Rho kinase inhibitor improved fibrosis and steatosis of the liver in CDAA-fed rats. The increase in the number of hepatocytes positive for 4-hydroxynonenal and 8 hydroxy-2'-deoxyguanosine in CDAA-fed rats was significantly prevented by the angiotensin II receptor blocker and the Rho kinase inhibitor. The levels of tumor necrosis factor-alpha mRNA in the liver of CDAA-fed rats were significantly increased and this increase was significantly inhibited by treatment with the angiotensin II receptor blocker and the Rho kinase inhibitor. mRNA expression of transforming growth factor-beta and alpha-smooth muscle actin stimulated by angiotensin II was also significantly suppressed by these two drugs. CONCLUSION: These results suggest that the Rho/Rho kinase pathway is at least partly involved in the renin-angiotensin system and plays an important role in hepatic fibrosis and steatosis. PMID- 17914986 TI - Education and imaging. Hepatobiliary and pancreatic: biliary tract opacification after angiography. PMID- 17914987 TI - Education and imaging. Hepatobiliary and pancreatic: emphysematous cholecystitis. PMID- 17914988 TI - Education and imaging. Gastrointestinal: emphysematous gastritis. PMID- 17914989 TI - Education and imaging. Gastrointestinal: enhanced gastric mucosal blood flow in aspirin intoxication. PMID- 17914990 TI - Education and imaging. Gastrointestinal: transdiaphragmatic intercostal hernia. PMID- 17914991 TI - Effect of a muscarinic M3 receptor agonist on gastric motility. AB - Muscarinic M3 receptors exist in the gastrointestinal wall in humans and the muscarinic M3 agonist cevimeline hydrochloride (Evoxac) is a candidate therapeutic agent for the treatment of xerostomia in Sjogren's syndrome. However, M3 receptor agonists are not known to show efficacy for diseases associated with abnormal gastrointestinal motility. Herein the effects are reported of cevimeline on gastric motility in two patients with non-ulcer dyspepsia. The patients both received long-term proton pump inhibitor therapy for 6 months, but their symptoms persisted. Then cevimeline was administered orally for 8 weeks at 30 mg three times daily (90 mg/day) and their dyspepsia symptoms improved. Electrogastrography was performed to examine gastric motility before and after administration of the M3 agonist. The fasting or nocturnal wave rate was significantly increased after administration compared with before administration, but no significant postprandial changes were seen. No adverse effects of cevimeline were observed. This drug might be a candidate therapeutic agent for non-ulcer dyspepsia. Because its postprandial effects on gastrointestinal motility are unclear, a dose-finding clinical study should be performed in the future. PMID- 17914992 TI - Hyperbaric oxygen therapy for severe acute pancreatitis. AB - Despite improvements in the supportive management of severe acute pancreatitis over the last decade, the morbidity and mortality rate remains high. The main feature of this condition is pancreatic necrosis leading to sepsis, with both localized and systemic inflammatory response syndromes. Early pathophysiological changes of the pancreas include alterations in microcirculation, ischemia reperfusion injury, and leukocyte and cytokine activation. The efficacy of hyperbaric oxygen (HBO) therapy in improving these pathophysiological disturbances is documented for various conditions. However, its effect in the treatment of severe acute pancreatitis is undetermined. This report documents the case of a 56-year-old woman presenting with severe acute pancreatitis treated by HBO therapy. The severity of disease was based on an Acute Physiology and Chronic Health Evaluation (APACHE II) illness grading score of 11 and a Baltazar based computed tomography severity index (CTSI) score of 9. Administration of 100% oxygen was commenced within 72 h of presentation at a pressure of 2.5 atmospheres for 90 min and given twice daily for a total of 5 days. Therapy was well tolerated with improvements in APACHE II and CTSI grading scores. HBO therapy for severe acute pancreatitis appeared to be safe and may have a role in improving treatment outcomes. Further study is required. PMID- 17914996 TI - Preschool psychopathology: lessons for the lifespan. PMID- 17914993 TI - Differentiation of metastatic follicular thyroid cancer from hepatocellular carcinoma using Hep Par 1. AB - Hepatocellular carcinoma usually arises in a cirrhotic liver. Multiple hepatic nodules in a non-cirrhotic liver are more likely to be metastatic. The primary focus commonly arises from the gastrointestinal tract, breast or lung, but in the absence of these a primary liver pathology must be considered. The case is reported of a middle-aged woman presenting with multiple nodules on computed tomography with no clinically apparent primary for whom results of initial diagnostic investigations were potentially misleading. PMID- 17914997 TI - Assessment of child problem behaviors by multiple informants: a longitudinal study from preschool to school entry. AB - BACKGROUND: Children's early problem behavior that manifests in multiple contexts is often more serious and stable. The concurrent and predictive validity of ratings of externalizing and internalizing by four informants was examined at preschool and early school age in an at-risk sample. METHODS: Two hundred forty children were assessed by mothers and fathers (Child Behavior Checklist (CBCL)), and teachers and laboratory examiners (Teacher Report Form (TRF)) at ages 3 and 5 years. RESULTS: All informants' ratings of externalizing converged on a common factor at ages 3 and 5 that showed strong stability over time (beta = .80). All informants' age 3 externalizing ratings significantly predicted the problem factor at age 5; mothers', fathers', and teachers' ratings were independently predictive. Ratings of internalizing (except by examiners at age 3) also converged at both ages; the problem factor showed medium stability (beta = .39) over time. Only fathers' ratings of age 3 internalizing predicted the age 5 problem factor. CONCLUSIONS: Findings support the value of multi-informant assessment, uphold calls to include fathers in childhood research, and suggest that examiners provide valid, though non-unique assessment data. Examiner contributions may prove useful in many research contexts. PMID- 17914998 TI - A developmental framework for distinguishing disruptive behavior from normative misbehavior in preschool children. AB - BACKGROUND: Attaining a developmentally sensitive nosology for preschool disruptive behavior requires characterization of the features that distinguish it from the normative misbehavior of this developmental period. We hypothesize that quality of behavior and its pervasiveness across contexts are critical dimensions for clinical discrimination in young children and propose that structured diagnostic observation provides a systematic method for their identification. We use the Disruptive Behavior Diagnostic Observation Schedule (DB-DOS) to examine whether: (a) observed quality and pervasiveness of behavior distinguishes preschoolers with clinically concerning disruptive behavior from typically developing preschoolers, and (b) observed pattern of clinically salient behavior predicts impairment above and beyond maternal report of behavioral frequency. METHODS: Participants are a behaviorally heterogeneous sample of preschoolers (N = 327). Diagnostic methods developed for clinical assessment of preschoolers were used to classify children as (a) Non-Disruptive, (b) Sub-Clinical, or (c) Disruptive. Child behavior was coded based on interactions with parent and examiner during the DB-DOS. RESULTS: Quality and pervasiveness of observed behaviors during the DB-DOS significantly distinguished the three behavioral groups. Discriminative utility varied depending on the comparison. With few exceptions, clinically concerning patterns on the DB-DOS added significant incremental utility in predicting impairment. CONCLUSIONS: Observed patterns of clinically salient behavior show promise for advancing developmentally-informed characterization of disruptive behavior within the preschool period. PMID- 17914999 TI - Gender differences in the behavioral associates of loneliness and social dissatisfaction in kindergarten. AB - BACKGROUND: Much of the evidence suggesting that loneliness is a risk factor for socio-emotional adjustment difficulties comes from studies with older children and adolescents. Comparatively less is known about the mental health implications of loneliness in early childhood. The goals of the present study were to provide additional convergent validity of the Loneliness and Social Dissatisfaction Questionnaire for Young Children (Cassidy & Asher, 1992) and explore potential gender differences in relations between loneliness and behavior problems in early childhood. METHODS: A multiple source assessment of 139 kindergarten-aged children (M(age) = 64.76 mos., SD = 4.48) was undertaken, including parental ratings, behavioral observations, child interviews, and teacher ratings. RESULTS: Overall, loneliness was positively associated with anxiety, aggression, and peer exclusion. However, several gender differences were also observed. CONCLUSIONS: Loneliness may be a marker variable for both early internalizing and externalizing problems. However, gender differences in the behavioral associates of loneliness are also evident. PMID- 17915000 TI - Associations between family relationships and symptoms/strengths at kindergarten age: what is the role of children's parental representations? AB - BACKGROUND: The quality of the family relationships plays an important role in the development of children's psychopathology and in their socio-emotional development. This longitudinal study aims to investigate whether family relationships are related to children's symptoms/strengths at kindergarten, and whether family relationships are predictors of changes in children's symptoms/strengths between 5 and 6. Our main interest is to examine the role of children's parental representations. METHODS: One hundred and fifty-three kindergarten children (67 girls, 86 boys) participated in this study. Children's parental representations were assessed using a story stem task (age 5). The family environment was assessed using parents' ratings (age 5). A multi-informant approach (parent, teacher, child) was employed to assess children's symptoms/strengths at 5 and 6. Children were interviewed using a standardised puppet interview. Parents and teachers completed questionnaires. RESULTS: Children's parental representations as well as the family environment were associated with children's symptoms/strengths at 5 and 6. When controlled for gender, children's representations were the only predictor of changes in symptoms/strengths between 5 and 6. A large number of negative parental representations at 5 predicted an increase in conduct problems. A large number of positive parental representations at 5 predicted an increase in pro-social behaviour. In terms of emotional symptoms and hyperactivity/inattention, symptoms at 5 were the only predictor for symptoms one year later. CONCLUSIONS: Our results suggest that kindergarten children's narrative representations may elaborate the processes underlying the expression of child disturbance and strengths, and underline the relevance of the play and narration of young children for diagnostics and therapy. PMID- 17915001 TI - Diagnostic specificity and nonspecificity in the dimensions of preschool psychopathology. AB - BACKGROUND: The appropriateness of the Diagnostic and Statistical Manual of Mental Disorders - Fourth Edition (DSM-IV) nosology for classifying preschool mental health disturbances continues to be debated. To inform this debate, we investigate whether preschool psychopathology shows differentiation along diagnostically specific lines when DSM-IV symptoms are aggregated statistically. METHODS: One thousand seventy-three parents of preschoolers aged 2-5 years attending a large pediatric clinic completed the Child Behavior Checklist 1.5-5. A stratified probability sample of 193 parents of high scorers and 114 parents of low scorers were interviewed with the Preschool Age Psychiatric Assessment (PAPA). Confirmatory factor analysis was performed on symptoms from seven DSM disorders. RESULTS: Comparison of competing models supported the differentiation of emotional syndromes into three factors: social phobia (SOC), separation anxiety (SAD), and depression/generalized anxiety (MDD/GAD), and the differentiation of disruptive syndromes into three factors: oppositional defiant/conduct syndrome (ODD/CD), hyperactivity/impulsivity, and inattention. Latent syndrome correlations were moderately high after accounting for symptom overlap and measurement error. CONCLUSIONS: Psychopathology appears to be differentiated among preschoolers much as it is among older children, and adolescents. We conclude that it is as reasonable to apply the DSM-IV nosology to preschoolers as it is to apply it to older individuals. PMID- 17915002 TI - Genetic and environmental influences on socio-emotional behavior in toddlers: an initial twin study of the infant-toddler social and emotional assessment. AB - BACKGROUND: Relatively little is known about the genetic architecture of childhood behavioral disorders in very young children. METHOD: In this study, parents completed the Infant-Toddler Social and Emotional Assessment, a questionnaire that assesses symptoms of childhood disorders, as well as socio emotional competencies, for 822 twin pairs (49.3% female; age 17-48 months) participating in the Wisconsin Twin Project. Psychometric, rater bias, and sex limitation models explored the role of genetic and environmental influences on (1) externalizing and internalizing behavior; (2) less commonly assessed behaviors pertaining to physical and emotional dysregulation, general competencies, social relatedness; and (3) infrequent behaviors such as those associated with pervasive developmental delays. RESULTS: Heritable influences accounted for the majority (56% or more) of variation in behavior that was commonly observed by both parents. The remaining variance was associated with non shared environmental factors, with the exception of competency and atypical behavior, which were also influenced by shared environmental factors. In contrast, for most behaviors, the variation unique to mother and father ratings was split between variation due to shared environment or rater biases and to measurement error. Little evidence emerged for sex differences in the underlying causes of variation. PMID- 17915003 TI - Positive and protective: effects of early theory of mind on problem behaviors in at-risk preschoolers. AB - BACKGROUND: Exposure to harsh parenting and children's skills in 'Theory of Mind' (ToM) show independent and interacting associations with problem behaviors at age 2 (Hughes & Ensor, 2006). This study examined whether these age-2 measures also predict age-4 problem behaviors. METHOD: In a socially diverse sample (N = 120), multi-informant, multi-measure, multi-setting ratings indexed problem behaviors at ages 2, 3 and 4; children completed both ToM and verbal-ability tasks at age 2, while video-based ratings of maternal negative affect and control within dyadic mother-child play indexed harsh parenting. RESULTS: Age-2 harsh parenting and ToM were independent and interacting predictors of age-4 problem behaviors, even with age-2 problem behaviors, verbal ability and social disadvantage all controlled. The interaction between harsh parenting and ToM distinguished persistent vs. diminishing problem behaviors. CONCLUSIONS: Both child and family characteristics predict increases in problem behaviors from 2 to 4; adverse effects of harsh parenting are attenuated for children with good ToM skills. PMID- 17915004 TI - Early concurrent and longitudinal symptoms of ADHD and ODD: relations to different types of inhibitory control and working memory. AB - BACKGROUND: The aim of the present study was to investigate how three different types of inhibitory control - interference control within task, interference control outside task, and prepotent response inhibition - and two types of working memory - verbal and spatial - would relate to early symptoms of attention deficit/hyperactivity disorder (ADHD) and oppositional defiant disorder (ODD), both concurrently and longitudinally. METHODS: Seventy-two preschoolers, 1/3 who had been identified as being at risk for developing ADHD and/or ODD, completed neuropsychological tasks designed to measure inhibitory control and working memory. Behavioral symptoms were measured through parental and teacher ratings of the DSM-IV criteria for ADHD and ODD. RESULTS: Our results suggest distinct types of inhibitory control as being good predictors of concurrent and longitudinal symptoms of ADHD, rather than ODD. However, no associations were obtained between working memory and ADHD or ODD symptoms either concurrently or longitudinally. CONCLUSIONS: This study emphasizes the need to isolate complex executive processes and break them down into components in order to properly understand the neuropsychological roots involved in ADHD and ODD. PMID- 17915005 TI - Unresolved maternal attachment representations, disrupted maternal behavior and disorganized attachment in infancy: links to toddler behavior problems. AB - BACKGROUND: Attachment theory's original formulation was substantially driven by Bowlby's (1969/1982) quest for a meaningful model of the development of psychopathology. Bowlby posited that aberrant experiences of parenting increase the child's risk of psychopathological outcomes, and that these risks are mediated by the quality of the attachment relationship. To empirically examine this hypothesis, the current study explores the associations between the development of toddler behavior problems and a) maternal unresolved attachment representations, b) maternal interactive behavior, and c) infant attachment relationships. Second, we test the mediating role of disorganized attachment in the association between disruptive behavior and toddler behavior problems, as well as unresolved attachment and behavior problems. METHOD: Sixty-four adolescent mother-infant dyads participated in this longitudinal study. The Adult Attachment Interview was administered at 6 months, the Strange Situation procedure was conducted at 12 months, disrupted behavior was assessed during play interactions at 12 months using the AMBIANCE measure, and the Child Behavior Checklist (CBCL) was used to assess behavior problems at 24 months of age. RESULTS: Maternal reports of externalizing problems were significantly associated with unresolved representations of attachment, disrupted maternal behavior, and disorganized attachment. Inclusion of these variables in a path analytic model suggested that disorganized attachment mediated the associations between disrupted maternal behavior and externalizing problems. Although the association between unresolved attachment representations and externalizing problems was no longer significant when mediation by disrupted behavior and disorganized attachment was taken into account, this indirect pathway was not significant. CONCLUSIONS: The results are consistent with Bowlby's (1969/1982) original conceptualization of the explanatory role of the attachment relationship in the development and manifestation of behavioral maladaptation. Effects of unresolved attachment on externalizing problems await further explanation. PMID- 17915006 TI - Hereditary sensory and autonomic neuropathies: types II, III, and IV. AB - The hereditary sensory and autonomic neuropathies (HSAN) encompass a number of inherited disorders that are associated with sensory dysfunction (depressed reflexes, altered pain and temperature perception) and varying degrees of autonomic dysfunction (gastroesophageal reflux, postural hypotention, excessive sweating). Subsequent to the numerical classification of four distinct forms of HSAN that was proposed by Dyck and Ohta, additional entities continue to be described, so that identification and classification are ongoing. As a group, the HSAN are rare diseases that affect both sexes. HSAN III is almost exclusive to individuals of Eastern European Jewish extraction, with incidence of 1 per 3600 live births. Several hundred cases with HSAN IV have been reported. The worldwide prevalence of HSAN type II is very low. This review focuses on the description of three of the disorders, HSAN II through IV, that are characterized by autosomal recessive inheritance and onset at birth. These three forms of HSAN have been the most intensively studied, especially familial dysautonomia (Riley-Day syndrome or HSAN III), which is often used as a prototype for comparison to the other HSAN. Each HSAN disorder is likely caused by different genetic errors that affect specific aspects of small fiber neurodevelopment, which result in variable phenotypic expression. As genetic tests are routinely used for diagnostic confirmation of HSAN III only, other means of differentiating between the disorders is necessary. Diagnosis is based on the clinical features, the degree of both sensory and autonomic dysfunction, and biochemical evaluations, with pathologic examinations serving to further confirm differences. Treatments for all these disorders are supportive. PMID- 17915007 TI - Analysis of gene order data supports vertical inheritance of the leukotoxin operon and genome rearrangements in the 5' flanking region in genus Mannheimia. AB - BACKGROUND: The Mannheimia subclades belong to the same bacterial genus, but have taken divergent paths toward their distinct lifestyles. For example, M. haemolytica + M. glucosida are potential pathogens of the respiratory tract in the mammalian suborder Ruminantia, whereas M. ruminalis, the supposed sister group, lives as a commensal in the ovine rumen. We have tested the hypothesis that vertical inheritance of the leukotoxin (lktCABD) operon has occurred from the last common ancestor of genus Mannheimia to any ancestor of the diverging subclades by exploring gene order data. RESULTS: We examined the gene order in the 5' flanking region of the leukotoxin operon and found that the 5' flanking gene strings, hslVU-lapB-artJ-lktC and xylAB-lktC, are peculiar to M. haemolytica + M. glucosida and M. granulomatis, respectively, whereas the gene string hslVU lapB-lktC is present in M. ruminalis, the supposed sister group of M. haemolytica + M. glucosida, and in the most ancient subclade M. varigena. In M. granulomatis, we found remnants of the gene string hslVU-lapB-lktC in the xylB-lktC intergenic region. CONCLUSION: These observations indicate that the gene string hslVU-lapB lktC is more ancient than the hslVU-lapB-artJ-lktC and xylAB-lktC gene strings. The presence of (remnants of) the ancient gene string hslVU-lapB-lktC among any subclades within genus Mannheimia supports that it has been vertically inherited from the last common ancestor of genus Mannheimia to any ancestor of the diverging subclades, thus reaffirming the hypothesis of vertical inheritance of the leukotoxin operon. The presence of individual 5' flanking regions in M. haemolytica + M. glucosida and M. granulomatis reflects later genome rearrangements within each subclade. The evolution of the novel 5' flanking region in M. haemolytica + M. glucosida resulted in transcriptional coupling between the divergently arranged artJ and lkt promoters. We propose that the chimeric promoter have led to high level expression of the leukotoxin operon which could explain the increased potential of certain M. haemolytica + M. glucosida strains to cause a particular type of infection. PMID- 17915008 TI - A study of the efficacy and cost-effectiveness of MRSA screening and monitoring on surgical wards using a new, rapid molecular test (EMMS). AB - BACKGROUND: MRSA is a significant contributor to prolonged hospital stay, poor clinical outcome and increased healthcare costs amongst surgical patients. A PCR test has been developed for rapid detection of MRSA in nasal swabs. The aims of this study are (1) to estimate the effectiveness of screening using this rapid PCR tests vs culture in reducing MRSA cross-infection rates; (2) to compare the cost of each testing strategy, including subsequent health care costs; and (3) to model different policies for the early identification and control of MRSA infection in surgical patients. METHODS/DESIGN: The study is a prospective two period cross-over study set in 7 surgical wards covering different surgical specialities. A total of 10,000 patients > 18 years will be tested over 16 months. The only difference between the two study periods is the method used for the detection of MRSA in each ward (rapid v conventional culture), with all other infection control practices remaining consistent between the arms. The study has been designed to complement routine practice in the NHS. Outcomes are MRSA cross infection rates (primary outcome) and need for antibiotic therapy and MRSA related morbidity. Parallel economic and modelling studies are being conducted to aid in the interpretation of the results and to evaluate the cost-effectiveness of the rapid PCR screening strategy. DISCUSSION: This paper highlights the design, methods and operational aspects of a study evaluating rapid MRSA screening in the surgical ward setting. PMID- 17915009 TI - Truncated recombinant human SP-D attenuates emphysema and type II cell changes in SP-D deficient mice. AB - BACKGROUND: Surfactant protein D (SP-D) deficient mice develop emphysema-like pathology associated with focal accumulations of foamy alveolar macrophages, an excess of surfactant phospholipids in the alveolar space and both hypertrophy and hyperplasia of alveolar type II cells. These findings are associated with a chronic inflammatory state. Treatment of SP-D deficient mice with a truncated recombinant fragment of human SP-D (rfhSP-D) has been shown to decrease the lipidosis and alveolar macrophage accumulation as well as production of proinflammatory chemokines. The aim of this study was to investigate if rfhSP-D treatment reduces the structural abnormalities in parenchymal architecture and type II cells characteristic of SP-D deficiency. METHODS: SP-D knock-out mice, aged 3 weeks, 6 weeks and 9 weeks were treated with rfhSP-D for 9, 6 and 3 weeks, respectively. All mice were sacrificed at age 12 weeks and compared to both PBS treated SP-D deficient and wild-type groups. Lung structure was quantified by design-based stereology at the light and electron microscopic level. Emphasis was put on quantification of emphysema, type II cell changes and intracellular surfactant. Data were analysed with two sided non-parametric Mann-Whitney U-test. MAIN RESULTS: After 3 weeks of treatment, alveolar number was higher and mean alveolar size was smaller compared to saline-treated SP-D knock-out controls. There was no significant difference concerning these indices of pulmonary emphysema within rfhSP-D treated groups. Type II cell number and size were smaller as a consequence of treatment. The total volume of lamellar bodies per type II cell and per lung was smaller after 6 weeks of treatment. CONCLUSION: Treatment of SP-D deficient mice with rfhSP-D leads to a reduction in the degree of emphysema and a correction of type II cell hyperplasia and hypertrophy. This supports the concept that rfhSP-D might become a therapeutic option in diseases that are characterized by decreased SP-D levels in the lung. PMID- 17915011 TI - A critical evaluation of loss of heterozygosity detected in tumor tissues, blood serum and bone marrow plasma from patients with breast cancer. AB - INTRODUCTION: The aim of the study was to perform a comparative analysis of LOH (loss of heterozygosity) in primary tumors as well as peripheral blood and bone marrow (BM) of patients with breast cancer (BCa). METHODS: Performing PCR-based fluorescence microsatellite analysis and using a panel of seven polymorphic microsatellite markers, we compared the profiles of LOH in primary tumors, peripheral blood and BM plasma from patients with primary BCa (n = 40, stage M0) as well as tumor tissues and blood serum from metastatic BCa patients (n = 48, stage M1). During the course of systemic treatment blood samplings from 12 M0 and 16 M1 patients were at least once repeated. RESULTS: The overall incidences of LOH in tumor tissues, blood and BM samples were 27.5%, 9.0% and 5.0%, respectively. The marker D3S1255 was the only one in the panel that showed similar frequencies of LOH ranging from 19.0 to 24.5% in tumor, blood and BM samples. Both M0 blood serum and BM plasma samples displayed the same rate of 19.0%, whereas tumor and M1 serum showed a rate of 24.5% and 24.0%, respectively, at this locus. This marker also showed the highest frequency of LOH in serum and BM samples, whereas in tumor samples LOHs at the markers D13S218 (38%) and D17S855 (36%) were more frequent. Statistical analysis of the tumor samples showed that occurrence of LOH at the markers D3S1255 (P < 0.04), D9S171 (P < 0.05) and D17S855 (P < 0.03) correlated with undifferentiated nuclear grade. Additionally, significant associations of the number of LOH recorded at D17S250 with estrogen receptor (P < 0.02), progesterone receptor (P < 0.03) expression and high proliferation score (Ki-67 expression, P = 0.009) were observed. In blood serum samples a relationship between positive lymph node status and LOH at the marker D3S1255 was revealed (M0 stage, P = 0.05; M0+M1 stage, P = 0.004). CONCLUSION: Our study demonstrates heterogeneous profiles and low rates of LOH, particularly on free DNA in BM and blood samples. However, the significant associations of LOH with some risk factors and the demonstrated possibility of monitoring free DNA in patients undergoing systemic therapy suggest that LOH analysis may be developed into a useful diagnostic tool. PMID- 17915010 TI - Genetic subtraction profiling identifies genes essential for Arabidopsis reproduction and reveals interaction between the female gametophyte and the maternal sporophyte. AB - BACKGROUND: The embryo sac contains the haploid maternal cell types necessary for double fertilization and subsequent seed development in plants. Large-scale identification of genes expressed in the embryo sac remains cumbersome because of its inherent microscopic and inaccessible nature. We used genetic subtraction and comparative profiling by microarray between the Arabidopsis thaliana wild-type and a sporophytic mutant lacking an embryo sac in order to identify embryo sac expressed genes in this model organism. The influences of the embryo sac on the surrounding sporophytic tissues were previously thought to be negligible or nonexistent; we investigated the extent of these interactions by transcriptome analysis. RESULTS: We identified 1,260 genes as embryo sac expressed by analyzing both our dataset and a recently reported dataset, obtained by a similar approach, using three statistical procedures. Spatial expression of nine genes (for instance a central cell expressed trithorax-like gene, an egg cell expressed gene encoding a kinase, and a synergid expressed gene encoding a permease) validated our approach. We analyzed mutants in five of the newly identified genes that exhibited developmental anomalies during reproductive development. A total of 527 genes were identified for their expression in ovules of mutants lacking an embryo sac, at levels that were twofold higher than in the wild type. CONCLUSION: Identification of embryo sac expressed genes establishes a basis for the functional dissection of embryo sac development and function. Sporophytic gain of expression in mutants lacking an embryo sac suggests that a substantial portion of the sporophytic transcriptome involved in carpel and ovule development is, unexpectedly, under the indirect influence of the embryo sac. PMID- 17915012 TI - Analysis and interpretation of dynamic FDG PET oncological studies using data reduction techniques. AB - BACKGROUND: Dynamic positron emission tomography studies produce a large amount of image data, from which clinically useful parametric information can be extracted using tracer kinetic methods. Data reduction methods can facilitate the initial interpretation and visual analysis of these large image sequences and at the same time can preserve important information and allow for basic feature characterization. METHODS: We have applied principal component analysis to provide high-contrast parametric image sets of lower dimensions than the original data set separating structures based on their kinetic characteristics. Our method has the potential to constitute an alternative quantification method, independent of any kinetic model, and is particularly useful when the retrieval of the arterial input function is complicated. In independent component analysis images, structures that have different kinetic characteristics are assigned opposite values, and are readily discriminated. Furthermore, novel similarity mapping techniques are proposed, which can summarize in a single image the temporal properties of the entire image sequence according to a reference region. RESULTS: Using our new cubed sum coefficient similarity measure, we have shown that structures with similar time activity curves can be identified, thus facilitating the detection of lesions that are not easily discriminated using the conventional method employing standardized uptake values. PMID- 17915013 TI - Finding coevolving amino acid residues using row and column weighting of mutual information and multi-dimensional amino acid representation. AB - BACKGROUND: Some amino acid residues functionally interact with each other. This interaction will result in an evolutionary co-variation between these residues - coevolution. Our goal is to find these coevolving residues. RESULTS: We present six new methods for detecting coevolving residues. Among other things, we suggest measures that are variants of Mutual Information, and measures that use a multidimensional representation of each residue in order to capture the physico chemical similarities between amino acids. We created a benchmarking system, in silico, able to evaluate these methods through a wide range of realistic conditions. Finally, we use the combination of different methods as a way of improving performance. CONCLUSION: Our best method (Row and Column Weighed Mutual Information) has an estimated accuracy increase of 63% over Mutual Information. Furthermore, we show that the combination of different methods is efficient, and that the methods are quite sensitive to the different conditions tested. PMID- 17915015 TI - Variations in brachial plexus and the relationship of median nerve with the axillary artery: a case report. AB - BACKGROUND: Brachial Plexus innervates the upper limb. As it is the point of formation of many nerves, variations are common. Knowledge of these is important to anatomists, radiologists, anesthesiologists and surgeons. The presence of anatomical variations of the peripheral nervous system is often used to explain unexpected clinical signs and symptoms. CASE PRESENTATION: On routine dissection of an embalmed 57 year old male cadaver, variations were found in the formation of divisions and cords of the Brachial Plexus of the right side. Some previously unreported findings observed were; direct branches to the muscles Pectoralis Minor and Latissimus dorsi from C6, innervation of deltoid by C6 and C7 roots and the origin of lateral pectoral nerve from the posterior division of upper trunk. The median nerve was present lateral to axillary artery. The left side brachial plexus was also inspected and found to have normal anatomy. CONCLUSION: The probable cause for such variations and their embryological basis is discussed in the paper. It is also concluded that although these variations may not have affected the functioning of upper limb in this individual, knowledge of such variations is essential in evaluation of unexplained sensory and motor loss after trauma and surgical interventions to the upper limb. PMID- 17915014 TI - Prevalence of self-reported diagnosis of diabetes mellitus and associated risk factors in a national survey in the US population: SHIELD (Study to Help Improve Early evaluation and management of risk factors Leading to Diabetes). AB - BACKGROUND: Studies derived from continuous national surveys have shown that the prevalence of diagnosed diabetes mellitus in the US is increasing. This study estimated the prevalence in 2004 of self-reported diagnosis of diabetes and other conditions in a community-based population, using data from the Study to Help Improve Early evaluation and management of risk factors Leading to Diabetes (SHIELD). METHODS: The initial screening questionnaire was mailed in 2004 to a stratified random sample of 200,000 households in the US, to identify individuals, age > or = 18 years of age, with diabetes or risk factors associated with diabetes. Follow-up disease impact questionnaires were then mailed to a representative, stratified random sample of individuals (n = 22,001) in each subgroup of interest (those with diabetes or different numbers of risk factors for diabetes). Estimated national prevalence of diabetes and other conditions was calculated, and compared to prevalence estimates from the National Health and Nutrition Examination Survey (NHANES) 1999-2002. RESULTS: Response rates were 63.7% for the screening, and 71.8% for the follow-up baseline survey. The SHIELD screening survey found overall prevalence of self-reported diagnosis of diabetes (either type 1 or type 2) was 8.2%, with increased prevalence with increasing age and decreasing income. In logistic regression modeling, individuals were more likely to be diagnosed with type 2 diabetes if they had abdominal obesity (odds ratio [OR] = 3.50; p < 0.0001), BMI > or =28 kg/m2 (OR = 4.04; p < 0.0001), or had been diagnosed with dyslipidemia (OR = 3.95; p < 0.0001), hypertension (OR = 4.82; p < 0.0001), or with cardiovascular disease (OR = 3.38; p < 0.0001). CONCLUSION: The SHIELD design allowed for a very large, community-based sample with broad demographic representation of the population of interest. When comparing results from the SHIELD screening survey (self-report only) to those from NHANES 1999-2002 (self-report, clinical and laboratory evaluations), the prevalence of diabetes was similar. SHIELD allows the identification of respondents with and without a current diagnosis of the illness of interest, and potential longitudinal evaluation of risk factors for future diagnosis of that illness. PMID- 17915016 TI - Caveolin-1 expression in benign and malignant lesions of the breast. AB - BACKGROUND: Caveolin-1 is thought to have an important impact on both signal transduction and mediation of intracellular processes. Furthermore, it has been suggested that Caveolin-1 may contribute to certain steps of carcinogenesis in various types of cancer. We examined the potential clinical relevance of Caveolin 1 in normal, benign and malignant breast tissue specimens. METHODS: Using tissue microarray (TMA) technology cases of invasive breast cancer, DCIS, benign breast disease (i.e. fibroadenoma, sclerosing adenosis, ductal hyperplasia and radial scar) and normal breast tissue were evaluated for Caveolin-1 expression. Immunohistochemical staining with an anti-Caveolin-1-antibody was performed. Staining intensity was quantified semiquantitatively. In invasive lesions staining results were correlated with clinical and pathological data. RESULTS: No Caveolin-1 expression was observed in epithelial cells of normal breast tissue (n = 5), benign breast disease (n = 295) and DCIS (n = 108). However, Caveolin-1 expression was found in 32 of 109 cases of invasive breast carcinomas (29.4%). Caveolin-1 expression in invasive breast cancer could neither be correlated with survival parameters such as overall or disease-free survival nor with established clinical and pathological markers. CONCLUSION: In this study we demonstrated expression of Caveolin-1 in one third of invasive breast cancers. A significant increase in Caveolin-1 expression was observed comparing invasive breast cancer to both benign breast tissue and non-invasive breast cancer. Since inhibitors of Caveolin-1 signalling are available, targeting Caveolin-1 in breast cancer may represent a potential option for future breast cancer treatment. PMID- 17915017 TI - Continuous control of endotracheal cuff pressure and tracheal wall damage: a randomized controlled animal study. AB - BACKGROUND: Intubation is frequently performed in intensive care unit patients. Overinflation of the endotracheal tube cuff is a risk factor for tracheal ischemia and subsequent complications. Despite manual control of the cuff pressure, overinflation of the endotracheal cuff is common in intensive care unit patients. We hypothesized that efficient continuous control of the endotracheal cuff pressure using a pneumatic device would reduce tracheal ischemic lesions in piglets ventilated for 48 hours through a high-volume, low-pressure endotracheal tube. MATERIALS AND METHODS: Twelve piglets were intubated and mechanically ventilated for 48 hours. Animals were randomized to manual control of the endotracheal cuff pressure (n = 6) or to continuous control of the endotracheal cuff pressure using a pneumatic device (n = 6). In the two groups, we inflated the endotracheal cuff with 50 ml air for 30 minutes, eight times daily. This hyperinflation of the endotracheal cuff aimed at mimicking high-pressure periods observed in intubated critically ill patients. In all animals, the cuff pressure and the airway pressure were continuously recorded for 48 hours. After sacrifice of the study animals, the trachea was removed and opened longitudinally for gross and histological examination. A pathologist evaluated the slides without knowledge of treatment group assignment. RESULTS: The cuff pressure was significantly lower in piglets with the pneumatic device than in piglets without the pneumatic device (median (interquartile range), 18.6 (11-19.4) cmH2O versus 26 (20-56) cmH2O, P = 0.009). No significant difference was found in the percentage of time spent with a cuff pressure <15 cmH2O and that with a cuff pressure between 30 and 50 cmH2O. The percentage of time between 15 and 30 cmH2O of cuff pressure, however, was significantly higher in piglets with the pneumatic device than in piglets without the pneumatic device (98% (95-99%) versus 65% (44 80%), P = 0.002). In addition, the percentage of time with cuff pressure >50 cmH2O was significantly lower in piglets with the pneumatic device than in piglets without the pneumatic device (0% versus 19% (12-41%), P = 0.002). In all animals, hyperemia and hemorrhages were observed at the cuff contact area. Histological examination showed no difference in tracheal lesions between animals with and without the pneumatic device. These lesions included deep mucous ulceration, squamous metaplasia and intense mucosal inflammation. No cartilage lesions were observed. CONCLUSION: The pneumatic device provided effective continuous control of high-volume, low-pressure endotracheal cuff pressure in piglets mechanically ventilated for 48 hours. In the present model, however, no significant difference was found in tracheal mucosal lesions of animals with or without a pneumatic device. Further studies are needed to determine the impact of continuous control of cuff pressure over a longer duration of mechanical ventilation. PMID- 17915018 TI - A systematic approach for testing expression of human full-length proteins in cell-free expression systems. AB - BACKGROUND: The growing field of proteomics and systems biology is resulting in an ever increasing demand for purified recombinant proteins for structural and functional studies. Here, we show a systematic approach to successfully express a full-length protein of interest by using cell-free and cell-based expression systems. RESULTS: In a pre-screen, we evaluated the expression of 960 human full length open reading frames in Escherichia coli (in vivo and in vitro). After analysing the protein expression rate and solubility, we chose a subset of 87 plasmids yielding no protein product in E. coli in vivo. These targets were subjected to a more detailed analysis comparing a prokaryotic cell-free E. coli system with an eukaryotic wheat germ system. In addition, we determined the expression rate, yield and solubility of those proteins. After sequence optimisation for the E. coli in vitro system and generating linear templates for wheat germ expression, the success rate of cell-free protein expression reached 93%. CONCLUSION: We have demonstrated that protein expression in cell-free systems is an appropriate technology for the successful expression of soluble full-length proteins. In our study, wheat germ expression using a two compartment system is the method of choice as it shows high solubility and high protein yield. PMID- 17915020 TI - Generalization through similarity: motif discourse in the discovery and elaboration of zinc finger proteins. AB - BACKGROUND: Biological organisms and their components are better conceived within categories based on similarity rather than on identity. Biologists routinely operate with similarity-based concepts such as "model organism" and "motif." There has been little exploration of the characteristics of the similarity-based categories that exist in biology. This study uses the case of the discovery and classification of zinc finger proteins to explore how biological categories based in similarity are represented. RESULTS: The existence of a category of "zinc finger proteins" was based in 1) a lumpy gradient of similarity, 2) a link between function and structure, 3) establishment of a range of appearance across systems and organisms, and 4) an evolutionary locus as a historically based common-ground. CONCLUSION: More systematic application of the idea of similarity based categorization might eliminate the assumption that biological characteristics can only contribute to narrow categorization of humans. It also raises possibilities for refining data-driven exploration efforts. PMID- 17915019 TI - Gene expression profiling of long-lived dwarf mice: longevity-associated genes and relationships with diet, gender and aging. AB - BACKGROUND: Long-lived strains of dwarf mice carry mutations that suppress growth hormone (GH) and insulin-like growth factor I (IGF-I) signaling. The downstream effects of these endocrine abnormalities, however, are not well understood and it is unclear how these processes interact with aging mechanisms. This study presents a comparative analysis of microarray experiments that have measured hepatic gene expression levels in long-lived strains carrying one of four mutations (Prop1(df/df), Pit1(dw/dw), Ghrhr(lit/lit), GHR-KO) and describes how the effects of these mutations relate to one another at the transcriptional level. Points of overlap with the effects of calorie restriction (CR), CR mimetic compounds, low fat diets, gender dimorphism and aging were also examined. RESULTS: All dwarf mutations had larger and more consistent effects on IGF-I expression than dietary treatments. In comparison to dwarf mutations, however, the transcriptional effects of CR (and some CR mimetics) overlapped more strongly with those of aging. Surprisingly, the Ghrhr(lit/lit) mutation had much larger effects on gene expression than the GHR-KO mutation, even though both mutations affect the same endocrine pathway. Several genes potentially regulated or co regulated with the IGF-I transcript in liver tissue were identified, including a DNA repair gene (Snm1) that is upregulated in proportion to IGF-I inhibition. A total of 13 genes exhibiting parallel differential expression patterns among all four strains of long-lived dwarf mice were identified, in addition to 30 genes with matching differential expression patterns in multiple long-lived dwarf strains and under CR. CONCLUSION: Comparative analysis of microarray datasets can identify patterns and consistencies not discernable from any one dataset individually. This study implements new analytical approaches to provide a detailed comparison among the effects of life-extending mutations, dietary treatments, gender and aging. This comparison provides insight into a broad range of issues relevant to the study of mammalian aging. In this context, 43 longevity associated genes are identified and individual genes with the highest level of support among all microarray experiments are highlighted. These results provide promising targets for future experimental investigation as well as potential clues for understanding the functional basis of lifespan extension in mammalian systems. PMID- 17915021 TI - Prior exercise and antioxidant supplementation: effect on oxidative stress and muscle injury. AB - BACKGROUND: Both acute bouts of prior exercise (preconditioning) and antioxidant nutrients have been used in an attempt to attenuate muscle injury or oxidative stress in response to resistance exercise. However, most studies have focused on untrained participants rather than on athletes. The purpose of this work was to determine the independent and combined effects of antioxidant supplementation (vitamin C + mixed tocopherols/tocotrienols) and prior eccentric exercise in attenuating markers of skeletal muscle injury and oxidative stress in resistance trained men. METHODS: Thirty-six men were randomly assigned to: no prior exercise + placebo; no prior exercise + antioxidant; prior exercise + placebo; prior exercise + antioxidant. Markers of muscle/cell injury (muscle performance, muscle soreness, C-reactive protein, and creatine kinase activity), as well as oxidative stress (blood protein carbonyls and peroxides), were measured before and through 48 hours of exercise recovery. RESULTS: No group by time interactions were noted for any variable (P > 0.05). Time main effects were noted for creatine kinase activity, muscle soreness, maximal isometric force and peak velocity (P < 0.0001). Protein carbonyls and peroxides were relatively unaffected by exercise. CONCLUSION: There appears to be no independent or combined effect of a prior bout of eccentric exercise or antioxidant supplementation as used here on markers of muscle injury in resistance trained men. Moreover, eccentric exercise as used in the present study results in minimal blood oxidative stress in resistance trained men. Hence, antioxidant supplementation for the purpose of minimizing blood oxidative stress in relation to eccentric exercise appears unnecessary in this population. PMID- 17915022 TI - Gene selection for classification of microarray data based on the Bayes error. AB - ABSTRACT: BACKGROUND: With DNA microarray data, selecting a compact subset of discriminative genes from thousands of genes is a critical step for accurate classification of phenotypes for, e.g., disease diagnosis. Several widely used gene selection methods often select top-ranked genes according to their individual discriminative power in classifying samples into distinct categories, without considering correlations among genes. A limitation of these gene selection methods is that they may result in gene sets with some redundancy and yield an unnecessary large number of candidate genes for classification analyses. Some latest studies show that incorporating gene to gene correlations into gene selection can remove redundant genes and improve classification accuracy. RESULTS: In this study, we propose a new method, Based Bayes error Filter (BBF), to select relevant genes and remove redundant genes in classification analyses of microarray data. The effectiveness and accuracy of this method is demonstrated through analyses of five publicly available microarray datasets. The results show that our gene selection method is capable of achieving better accuracies than previous studies, while being able to effectively select relevant genes, remove redundant genes and obtain efficient and small gene sets for sample classification purposes. CONCLUSION: The proposed method can effectively identify a compact set of genes with high classification accuracy. This study also indicates that application of the Bayes error is a feasible and effective wayfor removing redundant genes in gene selection. PMID- 17915023 TI - Patients' satisfaction and opinions of their experiences during admission in a tertiary care hospital in Pakistan - a cross sectional study. AB - BACKGROUND: It is often felt that developing countries need to improve their quality of healthcare provision. This study hopes to generate data that can help managers and doctors to improve the standard of care they provide in line with the wishes of the patients. METHODS: It was a cross sectional study carried out at a major tertiary care hospital of Karachi. Patients between the ages of 18 and 80 years admitted to the hospital for at least one day were included. Patients in the maternity, psychiatry and chemotherapy wards and those in the ICU/CCU were excluded. A pretested, peer reviewed translation of a validated patient satisfaction scale developed by the Picker Institute of Europe was administered. RESULTS: A total of 173 patients (response rate: 78.6 %) filled the questionnaire. Patient satisfaction was at levels comparable to European surveys for most aspects of hospital care. However, nearly half the patients (48%) felt they had to wait too long to get a bed in the hospital after presenting to the ER. 68.6% of the patients said that they were never asked for views on the quality of care provided. 20% of the patients did not find anyone in the staff to talk to about their worries and fears while 27.6% felt that they were given emotional support to only some extent. Up to one third of the patients said they were not provided enough information regarding their operative procedures beforehand. CONCLUSION: Although several components of patient care equal the quality levels of the west, many sections require considerable improvement in order to improve health care provision. The healthcare team needs to get more involved with the patients, providing them greater support and keeping them informed and involved with their medical treatment. Efforts should be made to get regular feedback from the patients. PMID- 17915024 TI - Mycobacterium tuberculosis 6-kDa early secreted antigenic target (ESAT-6) protein downregulates lipopolysaccharide induced c-myc expression by modulating the extracellular signal regulated kinases 1/2. AB - BACKGROUND: Mycobacterium tuberculosis (Mtb) causes death of 2-3 million people every year. The persistence of the pathogenic mycobacteria inside the macrophage occurs through modulation of host cell signaling which allows them, unlike the other non-pathogenic species, to survive inside the host. The secretory proteins of M. tuberculosis have gained attention in recent years both as vaccine candidates and diagnostic tools; they target the immune system and trigger a putatively protective response; however, they may also be involved in the clinical symptoms of the disease. RESULTS: Our studies showed that RD-1-encoded secretory protein ESAT-6 is involved in modulation of the mitogen-activated protein (MAP) kinase-signaling pathway inside the macrophage. ESAT-6 induced phosphorylation of extracellular signal-regulated kinases 1/2 (ERK1/2) in the cytoplasm but not in the nucleus, which normally is the case for MAP kinases. ESAT-6 also antagonized LPS-induced ERK1/2 phosphorylation in the nucleus. Stimulation of cells by ESAT-6 along with sodium orthovanadate (a tyrosine phosphatase inhibitor) restored phosphorylation of ERK1/2 in the nucleus, suggesting active dephosphorylation of ERK1/2 by some putative phosphatase(s) in the nucleus. Further, ESAT-6 was found to down regulate the expression of LPS inducible gene c-myc in an ERK1/2-dependent manner. CONCLUSION: This study showed the effect of secretory proteins of M. tuberculosis in the modulation of macrophage signaling pathways particularly ERK1/2 MAP kinase pathway. This modulation appears to be achieved by limiting the ERK1/2 activation in the nucleus which ultimately affects the macrophage gene expression. This could be a mechanism by which secretory proteins of Mtb might modulate gene expression inside the macrophages. PMID- 17915025 TI - Functional ability in a population: normative survey data and reliability for the ICF based Norwegian Function Assessment Scale. AB - BACKGROUND: The increasing focus on functional ability assessments in relation to sickness absence necessitates the measurement of population functional levels. This study assessed the reliability of the Norwegian Function Assessment Scale (NFAS) and presents normative population data. METHODS: All inhabitants in seven birth cohorts in Ullensaker municipality in 2004 were approached by means of a postal questionnaire. The NFAS was included as part of The Ullensaker Study 2004. The instrument comprises 39 items derived from the activities/participation component in the International Classification for Functioning, Disabilities and Health (ICF). Based on the results of principal component analysis, these items comprise seven domains. Non-parametric tests for independent samples were used to compare subgroups. Internal consistency was assessed by Cronbach's alpha. Two week test-retest reliability was assessed by total proportions of agreement, weighted kappa, and intraclass correlation coefficient (ICC). RESULTS: The response rate was 54% (1620 persons) and 75.4% (101 persons) for the retest. Items had low levels of missing data. Test-retest reliability was acceptable with high proportions of absolute agreement; kappa and ICC values ranged from 0.38 to 0.83 and 0.79 to 0.83, respectively. No difficulty on all 39 functional activities was reported by 33.1% of respondents. Females, older persons and persons with lower levels of education reported more functional problems than their respective counterparts (p < 0.05). The age gradient was most evident for three of the physical domains. For females aged 24-56 and males aged 44-76, a clear education gradient was present for three of the physical domains and one mental domain after adjusting for age and gender. CONCLUSION: This study presents population based normative data on functional ability, as measured by the NFAS. These data will serve as basis for the development of national population norms and are necessary for score interpretation. Data quality and test-retest reliability of the NFAS were acceptable. PMID- 17915026 TI - Assessment of free light chains in the cerebrospinal fluid of patients with lymphomatous meningitis - a pilot study. AB - BACKGROUND: Lymphomatous meningitis (LM) represents a severe complication of malignant lymphomas. While clinical suspicion is raised by symptoms ranging from mild disturbances of sensation to severe pain or impaired consciousness, the definite diagnosis of LM is often difficult to obtain. Since B-cell lymphomas are clonally restricted to express either kappa or lambda immunoglobulin light chain, we hypothesised that analysis of free light chain (FLC) ratios might facilitate the diagnosis of LM. METHODS: Kappa and lambda FLC were measured using a novel nephelometric assay in cerebrospinal fluid (CSF) and serum from 17 patients. 5/17 suffered from LM as demonstrated by cytology, immunocytology, and/or imaging procedures. RESULTS: Measurement of FLC concentrations in CSF was achieved for all 17 patients. FLC levels in CSF were lower than serum FLC levels in samples for the same patient obtained at the same time (p < 0.01). CSF and serum FLC concentrations correlated weakly in all patients irrespective of LM status. Significantly more patients with cytopathologically and immunohistochemically proven LM displayed abnormal kappa/lambda FLC ratios in CSF compared to individuals with no LM (p < 0.01). CONCLUSION: This is the first report demonstrating that a significant proportion of LM patients display an abnormal kappa/lambda FLC ratio in the CSF. PMID- 17915027 TI - Ecological correlates of sociality in Pemphigus aphids, with a partial phylogeny of the genus. AB - BACKGROUND: Because the systems of social organisation in the various species of Pemphigus aphids span the continuum from asociality through to advanced sociality (typified by the possession of morphologically specialised soldiers), the genus is an ideal model clade in which to study the influence of ecology on the origins of eusociality. We made detailed study of the ecology of three gall-dwelling species that show clear differences in their levels of social behaviour. To elucidate evolutionary relationships and to attempt to estimate the number of origins of sociality, we also created a phylogeny based on sequences spanning the mitochondrial genes Cytochrome Oxidase I and II for nine species of Pemphigus. RESULTS: P. spyrothecae, a highly social species with aggressive morphologically specialised soldiers, has the longest galling phase, unsynchronised development of a large number of individuals in a densely-populated gall, and an extended period over which alates emerge. P. populi, a species with no soldiers, has the shortest galling phase, synchronised development of a small number of individuals in a sparsely-populated gall, and an extremely brief emergence period. The ecology of P. bursarius, which has behavioural soldiers that are not morphologically specialised, is intermediate between these two extremes. The galls of P. spyrothecae and P. bursarius form small openings during the course of the season and predation-related mortality is relatively high in these two species. Conversely, predation does not occur during the galling phase of P. populi, which has no soldiers but makes an entirely-sealed gall. The phylogeny allowed us to infer one likely point of origin of basic social defence and two independent origins of enhanced defence. Based on current knowledge of behaviour, the phylogeny also suggests that the defence trait may have been lost at least once. CONCLUSION: The life-history strategy of P. spyrothecae appears to be geared towards defending the colony against the constant threat of predation that faces the inhabitants of a long-lived, open gall. The life-history strategy of P. populi, on the other hand, is to avoid predation in the closed gall fortress and flee for the secondary host at the earliest opportunity. The life-history strategy of P. bursarius appears to represent a compromise between these strategies. PMID- 17915028 TI - A cluster randomized trial evaluating electronic prescribing in an ambulatory care setting. AB - BACKGROUND: Medication errors, adverse drug events and potential adverse drug events are common and serious in terms of the harms and costs that they impose on the health system and those who use it. Errors resulting in preventable adverse drug events have been shown to occur most often at the stages of ordering and administration. This paper describes the protocol for a pragmatic trial of electronic prescribing to reduce prescription error. The trial was designed to overcome the limitations associated with traditional study design. DESIGN: This study was designed as a 65-week, cluster randomized, parallel study. METHODS: The trial was conducted within ambulatory outpatient clinics in an academic tertiary care centre in Ontario, Canada. The electronic prescribing software for the study is a Canadian electronic prescribing software package which provides physician prescription entry with decision support at the point of care. Using a handheld computer (PDA) the physician selects medications using an error minimising menu based pick list from a comprehensive drug database, create specific prescription instructions and then transmit the prescription directly and electronically to a participating pharmacy via facsimile or to the physician's printer using local area wireless technology. The unit of allocation and randomization is by 'week', i.e. the system is "on" or "off" according to the randomization scheme and the unit of analysis is the prescription, with adjustment for clustering of patients within practitioners. DISCUSSION: This paper describes the protocol for a pragmatic cluster randomized trial of point-of-care electronic prescribing, which was specifically designed to overcome the limitations associated with traditional study design. TRIAL REGISTRATION: This trial has been registered with clinicaltrials.gov (ID: NCT00252395). PMID- 17915029 TI - Interleukin-3 prevents neuronal death induced by amyloid peptide. AB - BACKGROUND: Interleukin-3 (IL-3) is an important glycoprotein involved in regulating biological responses such as cell proliferation, survival and differentiation. Its effects are mediated via interaction with cell surface receptors. Several studies have demonstrated the expression of IL-3 in neurons and astrocytes of the hippocampus and cortices in normal mouse brain, suggesting a physiological role of IL-3 in the central nervous system. Although there is evidence indicating that IL-3 is expressed in some neuronal populations, its physiological role in these cells is poorly known. RESULTS: In this study, we demonstrated the expression of IL-3 receptor in cortical neurons, and analyzed its influence on amyloid beta (Abeta)-treated cells. In these cells, IL-3 can activate at least three classical signalling pathways, Jak/STAT, Ras/MAP kinase and the PI 3-kinase. Viability assays indicated that IL-3 might play a neuroprotective role in cells treated with Abeta fibrils. It is of interest to note that our results suggest that cell survival induced by IL-3 required PI 3 kinase and Jak/STAT pathway activation, but not MAP kinase. In addition, IL-3 induced an increase of the anti-apoptotic protein Bcl-2. CONCLUSION: Altogether these data strongly suggest that IL-3 neuroprotects neuronal cells against neurodegenerative agents like Abeta. PMID- 17915030 TI - Extended analysis of benchmark datasets for Agilent two-color microarrays. AB - BACKGROUND: As part of its broad and ambitious mission, the MicroArray Quality Control (MAQC) project reported the results of experiments using External RNA Controls (ERCs) on five microarray platforms. For most platforms, several different methods of data processing were considered. However, there was no similar consideration of different methods for processing the data from the Agilent two-color platform. While this omission is understandable given the scale of the project, it can create the false impression that there is consensus about the best way to process Agilent two-color data. It is also important to consider whether ERCs are representative of all the probes on a microarray. RESULTS: A comparison of different methods of processing Agilent two-color data shows substantial differences among methods for low-intensity genes. The sensitivity and specificity for detecting differentially expressed genes varies substantially for different methods. Analysis also reveals that the ERCs in the MAQC data only span the upper half of the intensity range, and therefore cannot be representative of all genes on the microarray. CONCLUSION: Although ERCs demonstrate good agreement between observed and expected log-ratios on the Agilent two-color platform, such an analysis is incomplete. Simple loess normalization outperformed data processing with Agilent's Feature Extraction software for accurate identification of differentially expressed genes. Results from studies using ERCs should not be over-generalized when ERCs are not representative of all probes on a microarray. PMID- 17915031 TI - A pilot study on pupillary and cardiovascular changes induced by stereoscopic video movies. AB - BACKGROUND: Taking advantage of developed image technology, it is expected that image presentation would be utilized to promote health in the field of medical care and public health. To accumulate knowledge on biomedical effects induced by image presentation, an essential prerequisite for these purposes, studies on autonomic responses in more than one physiological system would be necessary. In this study, changes in parameters of the pupillary light reflex and cardiovascular reflex evoked by motion pictures were examined, which would be utilized to evaluate the effects of images, and to avoid side effects. METHODS: Three stereoscopic video movies with different properties were field-sequentially rear-projected through two LCD projectors on an 80-inch screen. Seven healthy young subjects watched movies in a dark room. Pupillary parameters were measured before and after presentation of movies by an infrared pupillometer. ECG and radial blood pressure were continuously monitored. The maximum cross-correlation coefficient between heart rate and blood pressure, rho max, was used as an index to evaluate changes in the cardiovascular reflex. RESULTS: Parameters of pupillary and cardiovascular reflexes changed differently after subjects watched three different video movies. Amplitudes of the pupillary light reflex, CR, increased when subjects watched two CG movies (movies A and D), while they did not change after watching a movie with the real scenery (movie R). The rho max was significantly larger after presentation of the movie D. Scores of the questionnaire for subjective evaluation of physical condition increased after presentation of all movies, but their relationship with changes in CR and rho max was different in three movies. Possible causes of these biomedical differences are discussed. CONCLUSION: The autonomic responses were effective to monitor biomedical effects induced by image presentation. Further accumulation of data on multiple autonomic functions would contribute to develop the tools which evaluate the effects of image presentation to select applicable procedures and to avoid side effects in the medical care and rehabilitation. PMID- 17915032 TI - Does Interpersonal Psychotherapy improve clinical care for adolescents with depression attending a rural child and adolescent mental health service? Study protocol for a cluster randomised feasibility trial. AB - BACKGROUND: Depression amongst adolescents is a costly societal problem. Little research documents the effectiveness of public mental health services in mapping this problem. Further, it is not clear whether usual care in such services can be improved via clinician training in a relevant evidence based intervention. One such intervention, found to be effective and easily learned amongst novice clinicians, is Interpersonal Psychotherapy (IPT). The study described in the current paper has two main objectives. First, it aims to investigate the impact on clinical care of implementing Interpersonal Psychotherapy for Adolescents for the treatment of adolescent depression within a rural mental health service compared with Treatment as Usual (TAU). The second objective is to record the process and challenges (i.e. feasibility, acceptability, sustainability) associated with implementing and evaluating an evidence-based intervention within a community service. This paper outlines the study rationale and design for this community based research trial. METHODS/DESIGN: The study involves a cluster randomisation trial to be conducted within a Child and Adolescent Mental Health Service in rural Australia. All clinicians in the service will be invited to participate. Participating clinicians will be randomised via block design at each of four sites to (a) training and delivery of IPT, or (b) TAU. The primary measure of impact on care will be a clinically significant change in depressive symptomatology, with secondary outcomes involving treatment satisfaction and changes in other symptomatology. Participating adolescents with significant depressive symptomatology, aged 12 to 18 years, will complete assessment measures at Weeks 0, 12 and 24 of treatment. They will also complete a depression inventory once a month during that period. This study aims to recruit 60 adolescent participants and their parent/guardian/s. A power analysis is not indicated as an intra-class correlation coefficient will be calculated and used to inform sample size calculations for subsequent large-scale trials. Qualitative data regarding process implementation will be collected quarterly from focus groups with participating clinicians over 18 months, plus phone interviews with participating adolescents and parent/guardians at 12 weeks and 24 weeks of treatment. The focus group qualitative data will be analysed using a Fourth Generation Evaluation methodology that includes a constant comparative cyclic analysis method. DISCUSSION: This study protocol will be informative for researchers and clinicians interested in considering, designing and/or conducting cluster randomised trials within community practice such as mental health services. TRIAL REGISTRATION: Australian Clinical Trials Registry ACTRNO12607000324415. PMID- 17915033 TI - Expression and water calcium dependence of calcium transporter isoforms in zebrafish gill mitochondrion-rich cells. AB - BACKGROUND: Freshwater fish absorb Ca2+ predominantly from ambient water, and more than 97% of Ca2+ uptake is achieved by active transport through gill mitochondrion-rich (MR) cells. In the current model for Ca2+ uptake in gill MR cells, Ca2+ passively enters the cytosol via the epithelium Ca2+ channel (ECaC), and then is extruded into the plasma through the basolateral Na+/Ca2+ exchanger (NCX) and plasma membrane Ca2+-ATPase (PMCA). However, no convincing molecular or cellular evidence has been available to support the role of specific PMCA and/or NCX isoforms in this model. Zebrafish (Danio rerio) is a good model for analyzing isoforms of a gene because of the plentiful genomic databases and expression sequence tag (EST) data. RESULTS: Using a strategy of BLAST from the zebrafish genome database (Sanger Institute), 6 isoforms of PMCAs (PMCA1a, PMCA1b, PMCA2, PMCA3a, PMCA3b, and PMCA4) and 7 isoforms of NCXs (NCX1a, NCX1b, NCX2a, NCX2b, NCX3, NCX4a, and NCX4b) were identified. In the reverse-transcriptase polymerase chain reaction (RT-PCR) analysis, 5 PMCAs and 2 NCXs were ubiquitously expressed in various tissues including gills. Triple fluorescence in situ hybridization and immunocytochemistry showed the colocalization of zecac, zpmca2, and zncx1b mRNAs in a portion of gill MR cells (using Na+-K+-ATPase as the marker), implying a subset of ionocytes specifically responsible for the transepithelial Ca2+ uptake in zebrafish gills. The gene expressions in gills of high- or low-Ca2+-acclimated zebrafish by quantitative real-time PCR analysis showed that zecac was the only gene regulated in response to environmental Ca2+ levels, while zpmcas and zncxs remained steady. CONCLUSION: The present study provides molecular evidence for the specific isoforms of Ca2+ transporters, zECaC, zPMCA2, and zNCX1b, supporting the current Ca2+ uptake model, in which ECaC may play a role as the major regulatory target for this mechanism during environmental challenge. PMID- 17915034 TI - Finding disease candidate genes by liquid association. AB - A novel approach to finding candidate genes by using gene expression data through liquid association is developed and used to identify multiple sclerosis susceptibility candidate genes. PMID- 17915035 TI - Protocol for Fit Bodies, Fine Minds: a randomized controlled trial on the affect of exercise and cognitive training on cognitive functioning in older adults. AB - BACKGROUND: Declines in cognitive functioning are a normal part of aging that can affect daily functioning and quality of life. This study will examine the impact of an exercise training program, and a combined exercise and cognitive training program, on the cognitive and physical functioning of older adults. METHODS/DESIGN: Fit Bodies, Fine Minds is a randomized, controlled trial. Community-dwelling adults, aged between 65 and 75 years, are randomly allocated to one of three groups for 16 weeks. The exercise-only group do three 60-minute exercise sessions per week. The exercise and cognitive training group do two 60 minute exercise sessions and one 60-minute cognitive training session per week. A no-training control group is contacted every 4 weeks. Measures of cognitive functioning, physical fitness and psychological well-being are taken at baseline (0 weeks), post-test (16 weeks) and 6-month follop (40 weeks). Qualitative responses to the program are taken at post-test. DISCUSSION: With an increasingly aged population, interventions to improve the functioning and quality of life of older adults are particularly important. Exercise training, either alone or in combination with cognitive training, may be an effective means of optimizing cognitive functioning in older adults. This study will add to the growing evidence base on the effectiveness of these interventions. TRIAL REGISTRATION: Australian Clinical Trials Register: ACTRN012607000151437. PMID- 17915036 TI - Phylogeography and domestication of Indian river buffalo. AB - BACKGROUND: The water buffalo- Bubalus bubalis holds tremendous potential in livestock sector in many Asian countries, particularly India. The origin, domestication and genetic structure of the Indian river buffalo are poorly understood. Therefore, to understand the relationship among the maternal lineages of Indian river buffalo breeds and their domestication process, we analysed mitochondrial D-loop region of 217 animals representing eight breeds from eight different locations in India along with published sequences of Mediterranean buffalo. RESULTS: The maximum parsimony tree showed one major clade with six internal branches. Reduced median network revealed expansion from more than one set of haplotypes indicating complex domestication events for this species. In addition, we found several singleton haplotypes. Using rho statistics, we obtained a time estimate of 6300 years BP for the expansion of one set of hapltoypes of the Indian domestic buffalo. A few breed specific branches in the network indicated an ancient time depth of differentiation of some of the maternal lineages of river buffalo breeds. The multidimensional display of breed pairwise FST values showed significant breed differentiation. CONCLUSION: Present day river buffalo is the result of complex domestication processes involving more than one maternal lineage and a significant maternal gene flow from the wild populations after the initial domestication events. Our data are consistent with the available archaeological information in supporting the proposition that the river buffalo was likely to be domesticated in the Western region of the Indian subcontinent, specifically the present day breeding tracts of the Mehsana, Surati and Pandharpuri breeds. PMID- 17915038 TI - Extended homozygosity is not usually due to cytogenetic abnormality. AB - BACKGROUND: Previous studies have reported frequent stretches of homozygosity in human subjects but have failed to clarify whether these are due to cytogenetic abnormalities or to autozygosity. METHODS: Trios which had been typed for closely spaced SNPs spanning the genome were studied. Stretches of extended homozygosity were identified in the child members, as were occasions on which the child had been genotyped as not inheriting one parental allele. The number of times such transmission errors occurred within regions of extended homozygosity was compared with the chance expectation. RESULTS: Transmission errors occurred more rarely in regions of extended homozygosity than would be expected by chance. DISCUSSION: Regions of extended homozygosity are not generally due to cytogenetic abnormalities such as uniparental isodisomy. They reflect the Mendelian inheritance of haplotypes from a common ancestor. This may have implications for mapping disease genes. PMID- 17915039 TI - Strange days. PMID- 17915037 TI - An unusual S-adenosylmethionine synthetase gene from dinoflagellate is methylated. AB - BACKGROUND: S-Adenosylmethionine synthetase (AdoMetS) catalyzes the formation of S-Adenosylmethionine (AdoMet), the major methyl group donor in cells. AdoMet mediated methylation of DNA is known to have regulatory effects on DNA transcription and chromosome structure. Transcription of environmental-responsive genes was demonstrated to be mediated via DNA methylation in dinoflagellates. RESULTS: A full-length cDNA encoding AdoMetS was cloned from the dinoflagellate Crypthecodinium cohnii. Phylogenetic analysis suggests that the CcAdoMetS gene, is associated with the clade of higher plant orthrologues, and not to the clade of the animal orthrologues. Surprisingly, three extra stretches of residues (8 to 19 amino acids) were found on CcAdoMetS, when compared to other members of this usually conserved protein family. Modeled on the bacterial AdeMetS, two of the extra loops are located close to the methionine binding site. Despite this, the CcAdoMetS was able to rescue the corresponding mutant of budding yeast. Southern analysis, coupled with methylation-sensitive and insensitive enzyme digestion of C. cohnii genomic DNA, demonstrated that the AdoMetS gene is itself methylated. The increase in digestibility of methylation-sensitive enzymes on AdoMet synthetase gene observed following the addition of DNA methylation inhibitors L ethionine and 5-azacytidine suggests the presence of cytosine methylation sites within CcAdoMetS gene. During the cell cycle, both the transcript and protein levels of CcAdoMetS peaked at the G1 phase. L-ethionine was able to delay the cell cycle at the entry of S phase. A cell cycle delay at the exit of G2/M phase was induced by 5-azacytidine. CONCLUSION: The present study demonstrates a major role of AdoMet-mediated DNA methylation in the regulation of cell proliferation and that the CcAdoMetS gene is itself methylated. PMID- 17915040 TI - Complexity and integration in the control of inner-ear development. PMID- 17915041 TI - Projections of the nucleus of the basal optic root in pigeons (Columba livia): a comparison of the morphology and distribution of neurons with different efferent projections. AB - The avian nucleus of the basal optic root (nBOR) is a visual structure involved in the optokinetic response. nBOR consists of several morphologically distinct cell types, and in the present study, we sought to determine if these different cell types had differential projections. Using retrograde tracers, we examined the morphology and distribution of nBOR neurons projecting to the vestibulocerebellum (VbC), inferior olive (IO), dorsal thalamus, the pretectal nucleus lentiformis mesencephali (LM), the contralateral nBOR, the oculomotor complex (OMC) and a group of structures along the midline of the mesencephalon. The retrogradely labeled neurons fell into two broad categories: large neurons, most of which were multipolar rather than fusiform and small neurons, which were either fusiform or multipolar. From injections into the IO, LM, contralateral nBOR, and structures along the midline-mesencephalon small nBOR neurons were labeled. Although there were no differences with respect to the size of the labeled neurons from these injections, there were some differences with the respect to the distribution of labeled neurons and the proportion of multipolar vs. fusiform neurons. From injections into the VbC, the large multipolar cells were labeled throughout nBOR. The only other cases in which these large neurons were labeled were contralateral OMC injections. To investigate if single neurons project to multiple targets we used paired injections of red and green fluorescent retrograde tracers into different targets. Double-labeled neurons were never observed indicating that nBOR neurons do not project to multiple targets. We conclude that individual nBOR neurons have unique projections, which may have differential roles in processing optic flow and controlling the optokinetic response. PMID- 17915042 TI - Eye dominance and response latency in area V1 of the monkey. AB - We measured the latency of 35 cells from V1 in two rhesus monkeys, to dynamic random dot stimuli monocular and binocularly presented. Mean latencies after non dominant eye stimulation (97.9 ms) were longer than those for dominant eye (78.2 ms) and binocular (70.7 ms) stimulation. Differences between latencies for dominant eye and binocular stimulation were not statistically significant. For dominant eye, there was a significant statistical correlation between dominance strength and latency (R = -0.36; p = 0.03). We failed to find significant statistical differences between latencies for cells with temporal and nasal dominant receptive-field. We conclude that, in V1, the response latency is largely determined by the dominant eye, whereas interocular interactions do not seem to play a relevant role regarding response latency. PMID- 17915043 TI - Cytoskeleton alteration correlates with gross structural plasticity in the cat lateral geniculate nucleus. AB - Monocular deprivation during early development causes rearrangement of neural connections within the visual cortex that produces a shift in ocular dominance favoring the non-deprived eye. This alteration is manifested anatomically within deprived layers of the lateral geniculate nucleus (LGN) where neurons have smaller somata and reduced geniculocortical terminal fields compared to non deprived counterparts. Experiments using monocular deprivation have demonstrated a spatial correlation between cytoskeleton alteration and morphological change within the cat LGN, raising the possibility that subcellular events mediating deprivation-related structural rearrangement include modification to the neuronal cytoskeleton. In the current study we compared the spatial and temporal relationships between cytoskeleton alteration and morphological change in the cat LGN. Cross-sectional soma area and neurofilament labeling were examined in the LGN of kittens monocularly deprived at the peak of the critical period for durations that ranged from 1 day to 7 months. After 4 days of deprivation, neuron somata within deprived layers of the LGN were significantly smaller than those within non-deprived layers. This structural change was accompanied by a spatially coincident reduction in neurofilament immunopositive neurons that was likewise significant after 4 days of deprivation. Both anatomical effects reached close to their maximum by 10 days of deprivation. Results from this study demonstrate that alteration to the neuronal cytoskeleton is both spatially and temporally linked to the gross structural changes induced by monocular deprivation. PMID- 17915044 TI - Social value orientation and cooperation in social dilemmas: a review and conceptual model. AB - Social psychologists have long recognized that people fundamentally differ with respect to their social value orientation (SVO), that is, self-regarding versus other regarding preferences, and that these differences affect cooperative behaviour in situations of interdependence. In this paper, we systematically review the vast number of findings on SVO and cooperation, and synthesize the state of the art by presenting an integrated conceptual model that may explain why and when people with different social values select different behavioural strategies in social dilemmas. Specifically, building on Pruitt and Kimmel's (1977) goal/expectation theory and our review of the literature, we suggest that the relationship between SVO and cooperative behaviour is mediated by (1) a cooperative goal and (2) the specific expectations concerning alters' behaviour. We also propose that trust and goal alignment are important contextual moderators of this relationship: for prosocials, cues signalling trust are necessary to generate positive expectations regarding alters' behaviour, whereas proselfs need external incentives to align their personal interest with a cooperative goal. We conclude this review by pointing to several avenues for future research that would help to deepen our understanding of the role of SVO in human cooperation. PMID- 17915045 TI - Identification and early characterization of genetically modified NGF-producing neural stem cells grafted into the injured adult rat brain. AB - OBJECTIVE: To investigate whether genetically modified mouse neural stem cells (NSC) expressing recombinant human nerve growth factor (rhNGF) and transplanted in chemically injured rat brain, can survive and eventually acquire phenotypic characteristics of early nerve cells. METHODS: Stably high expression of rhNGF in NSC was obtained by a new lentivirus-mediated expression system. To test the effectiveness of hNGF secreted by rhNGF-NSC, hereby we performed either a bioassay for neurite outgrowth in PC12 rat cells or immunoblot analysis for TrkA, the high-affinity NGF receptor, from engineered NSC. rhNGF and mock-NSC were grafted into adult injured rats striatum and 3 days later, animals were killed, and brains were removed and examined by immunohistochemical analysis. RESULTS: The results showed that rhNGF-producing NSC cultured for extended period of time release bioactive hNGF in the culture media which promotes PC12 neuronal differentiation and correlates with the up-regulation of TrkA. rhNGF-NSC transplanted into the injured brain can survive, produce hNGF and induce the expression of NGF receptors, p75(NTR) and TrkA. DISCUSSION: In vitro and in vivo experiments confirmed the ability of rhNGF-NSC to secrete bioactive hNGF. Our data provide by means of genetically modified rhNGF-producing NSC, a useful experimental tool to test the potential clinical effectiveness of trophic factors relevant to central nervous system (CNS). PMID- 17915046 TI - Characteristics of the locomotor-like muscle activity during orthotic gait in paraplegic persons. AB - We previously found that orthotic gait training can induce 'locomotor-like' coordinated muscle activity of the paralyzed lower limb in persons with spinal cord injury (SCI). The purpose of the present study was to characterize the locomotor-like muscle activity based on data obtained from electromyographic recordings and motion analysis during orthotic gait in nine motor complete SCI subjects. Seven of nine subjects showed a common EMG activation pattern mainly in the ankle (soleus: Sol) and hip extensor (biceps femoris: BF) muscles. The locomotor-like muscle activity was well synchronized with the gait cycle, namely, the EMG amplitude of both Sol and BF muscle had common temporal relationships with the ground reaction force, and hip and ankle joint motions. While the presence or absence of the EMG activity during orthotic gait was consistent with those of mechanically-induced stretch reflex, the duration and amount area of the locomotor-like muscle activity were significantly longer than those of the stretch reflex in the Sol muscle. Moreover, the Sol EMG magnitude had strong relevance to hip as well as ankle angular velocities. These results indicate that the locomotor-like muscle activity during orthotic gait is not a mere reflex response, but includes an activity of the central pattern generator (CPG) and its interaction with afferent inputs. Orthotic gait training for complete SCI persons might have a potential to activate the spinal locomotor center. PMID- 17915048 TI - [Traumatic retroperitoneal haematoma]. AB - The management of traumatic retroperitoneal haematomas is still a much debated question. Although the diagnosis has become easier using CT with contrast medium, the therapeutic decisions are still difficult because of the great variability of the lesions, which may be simple but very often complicated. Our study is based on 1086 treated patients, 29.5% of the 3682 critical abdominal polytrauma seen in 35 years. Mortality has been 12.9% with a medium ISS (Injury Severity Score) of 23.4. 71.4% of the cases were closed traumas, 28,6% were open traumas. The most common single lesions have been pelvic (43%), followed by the renal traumas (39%). Regarding the associated lesions, the thoracic traumas cause an increment of the ISS score up to 26.2% and of mortality up to 14.6%. The maxillofacial traumas associated with traumatic retroperitoneal haematomas represent 11%, mainly associated with motorcycle accidents, which have increased in the last years from 2,4% in the 70s to 32% these days. Our approach to these patients has been basically conservative. Following the indications obtained by the CT, we widely used interventional angiography, especially for renal lesions and, after pelvic stabilization, for pelvic haematomas. We have chosen surgery considering the kind of traumas (open or closed), the location of the haematoma and especially, the clinical course of the patient's hemodynamic condition. PMID- 17915047 TI - [Single thyroid nodule]. PMID- 17915049 TI - Endoscopic treatment of bilateral pheochromocytoma in MEN 2A syndrome: case report and review of the literature. AB - The benefits of laparoscopic adrenalectomy for single adrenal lesion have been well documented in literature; less experience though has been reported with simultaneous bilateral laparoscopic adrenalectomy. This operation is indicated in case of primary hypercortisolism caused by bilateral adrenocortical hyperfunction, Cushing's disease after failure of pituitary surgery, ectopic adrenocorticotropic hormone (ACTH) production by a tumour inaccessible for surgical intervention, and pheochromocytoma when it occurs bilaterally in case of multiple endocrine neoplasia type 2A and 2B. Different laparoscopic approaches have been described to perform this operation, such as the "anterior" approach (transperitoneal), the "lateral" approach (transperitoneal and retroperitoneal), and the "posterior" approach (retroperitoneal). We report a case of bilateral laparoscopic adrenalectomy in a 33 years old female affected with bilateral pheochromocytoma due to multiple endocrine neoplasia type 2A treated with a bilateral transperitoneal laparoscopic adrenalectomy and disease free after 18 months follow-up. PMID- 17915050 TI - Combined resection and multi-agent adjuvant chemotherapy for intra-abdominal desmoplastic small round cell tumour: case report and review of the literature. AB - Desmoplastic small round cell tumor (DSRCT) is a rare, highly aggressive malignancy with distinctive histological and immunohistochemical features occurring in young population with male predominance. We report a case of DRSCT occurred in a 17 years old patient which presented with a large upper left quadrant abdominal mass that was treated with a very aggressive surgical approach and multi-agent chemotherapy. At a 12 months follow-up he is free of recurrence. This kind of tumour has a very poor prognosis. No standard treatment protocol has been established. Aggressive surgery combined with postoperative multi-agent adjuvant chemotherapy is justified not only to relieve symptoms but also to try to improve the outcome. PMID- 17915051 TI - [Adenocarcinoma following restorative proctocolectomy for ulcerative colitis: a case report and review of the literature]. AB - Restorative proctocolectomy with ileal pouch-anal anastomosis has been accepted as the surgical treatment of choice for most patients with ulcerative colitis. The occurrence of adenocarcinoma arising near or into the ileal pouch is rare. Only 19 such cases have been reported so far. The authors report a case of a 67 year old male who developed an adenocarcinoma in the small rectal stump 12 years after a restorative proctocolectomy with double stapled ileal pouch-low rectal anastomosis for ulcerative colitis unresponsive to medical treatment. They, after a literature review, examine same steps of the procedure and emphasize the importance of regular and prolonged follow-up for all patients having restorative proctocolectomy for ulcerative colitis. PMID- 17915052 TI - [Spontaneous renal bleeding in a dialysis patient with acquired cystic disease of the kidney: case report and review of the literature]. AB - The aim of this paper is to describe a rare clinical case of spontaneous haemorrhagic rupture of a multicystic kidney in a patient on haemodialysis for acquired cystic disease. We also discuss current issues about the management of this rare condition, with a short review of the literature. PMID- 17915053 TI - [Tenosynovial giant cell tumour of the hand and wrist: early diagnosis and surgical treatment]. AB - Fourty two patients with tenosynovial giant cell tumour, treated with radical excision, underwent detailed follow-up to evaluate the outcomes and to find early the possible recurrence of disease. Radical excision is the only way to have a complete recovery from disease and total recovery of function. Early diagnosis of recurrence is possible by careful clinical examination, ultrasonography and, in selected cases, magnetic resonance imaging. PMID- 17915054 TI - [Surgical management of medullary thyroid carcinoma. Personal experience]. AB - INTRODUCTION: Medullary Thyroid Carcinoma (MTC) is a rare malignancy of the parafollicular C cells of the thyroid gland. Aim of our study is reporting retrospective analysis of our experience about the surgical treatment of MTC, especially pointing on lymphadenectomy. PATIENTS AND METHODS: from January 2000 to March 2006, were performed 546 thyroidectomy, 6 of them for MTC (1.09%): 5 sporadic and 1 familiar. All the patients were operated with standard technique (total thyroidectomy) and submitted to endocrinologic and instrumental follow-up. RESULTS: in our experience, we do not registered early or late complications as bleeding, laryngeal nerves lesions and parathyroid lesions or intra-postoperative deaths. DISCUSSION: MTC occurs sporadically or as part of the multiple endocrine neoplasia type 2 (MEN 2 a/b) syndromes in patients who have inherited a mutation in the RET proto-oncogene. The diagnosis is made by fine needle aspiration biopsy (FNAB) and by measuring calcitonin levels in the blood. Primary treatment consists of surgical resection including a total thyroidectomy, central neck nodal dissection and functional lateral neck nodal dissections. Most patients with a palpable primary tumour have nodal disease at the time of operation, and nodal involvement is often bilateral. CONCLUSIONS: adequate resection of the primary tumour and cervical lymph nodes is important to optimize outcome and minimize the risk of recurrent disease. Following primary surgical resection, more than 50% of the patients will have recurrent disease with persistent elevation of calcitonin levels. Currently, there is no adequate systemic therapy for recurrent disease. Surgical reoperation or conservative observation are the best available options. PMID- 17915055 TI - [Surgical timing in bleeding liver adenoma: case report]. AB - The diagnosis of liver adenoma, which etiopathogenesis most often involves a prolonged assumption of estrogen (90% of adenomas occurs in women after more than 5 years of estrogen therapy), always imposes a surgical resection. The reason depend from neoplasia characteristics like the malignant evolution (4%) and the high risk of abdominal/intratumoral bleeding (30-50%), that increases during pregnancy and postpartum period. Regression of lesion after discontinuation of hormone therapy is rare and does not remove the degeneration and/or haemorrhagic risk. Liver resection should be performed with appropriate selective endovascular embolization, considering that an inept emergency surgery may impose a greater risk ot the liver, exposing the patient to major risk of morbidity and mortality. The correct timing from embolization to elective surgery is not yet standardized in the literature. The surgeon's personal experience and mainly a careful patient follow-up suggest the timing of surgery after embolization. The authors relate their own experience about the therapeutic strategy and surgical timing in a case of bleeding liver adenoma. PMID- 17915056 TI - [Electronic sheet in the management of difficult wounds]. AB - Nowadays, the treatment of the difficult wounds represents an emergent socio sanitary problem, due to the increase of the average duration of life, with consequent increasing costs by the National Sanitary System. The term 'difficult wounds' refers to all losses of cutaneous substances with multifactorial pathogenesis that do not spontaneously recover. Today it is possible to use advanced dressings, representing a valid tool to speed-up the healing process that--as a consequence--improves quality of life for the patient. These patients need to be followed by medical teams composed by different specialists sometimes working in different hospitals. As a consequence it is has been necessary to create an electronic document containing the clinical history of the patient and reporting the different treatments. The electronic sheet allows: a) to evaluate in detail the evolution of patient conditions, thanks to an always available iconographic documentation, even when the patient is not followed by the same physician; b) and to test the effectiveness of the new advanced dressings available on the market. PMID- 17915057 TI - [The role of colonoscopy in patient follow-up after surgery for colorectal cancer. A retrospective study and review of the literature]. AB - To improve survival rate after colon or rectum resection for cancer patients should be strictly followed up in order to identify possible local disease relapse or metachronous neoplasia. From October 2002 to January 2006, 864 patients had undergone colonoscopy and 68 were treated surgically for colorectal adenocarcinoma. Of these, 36 were men and 32 women, with a mean age of 63 years. Nineteen of the patients underwent a left colectomy, 28 an anterior resection, 18 a right colectomy and 3 a resection of the transverse colon. For all these patients follow-up program include a colonoscopy performed annually for the first two years, and subsequently, if the results were negative, after a further three and then five years. Out of 68 patients, 2 showed suspect anastomotic recurrence, which proved to be granulomas at the histological examination. In addition, in 11 cases, there were 3 right colon adenomatous polyps, 2 transverse colon polyps (one villous and the other tubular), 5 descending colon polyps (three tubular and two villous) and 1 tubulo-villous polyp of the rectum. No metachronous neoplasias were observed. An examination of the data resulting from our own 68 cases shows that, in spite of the fact that no local disease relapse or metachronous neoplasia was observed, the identification of 11 polyps would suggest that the use of colonoscopy in such patients might be the gold standard for early diagnosis of recurrences and new polyps. PMID- 17915059 TI - Severe Asherman's syndrome. PMID- 17915058 TI - Laparoscopic treatment of splenic lymphangiomas: report of three cases. AB - BACKGROUND AND AIM: Since 1991, laparoscopic splenectomy has been performed in many different pathologies of the spleen. Although it is a rare lesion, splenic lymphangiomas are cystic lesions of the spleen requiring splenectomy. Herein, we present three females who have undergone laparoscopic splenectomy with the diagnosis of cystic splenic lymphangioma. PATIENTS AND METHODS: In the last four years, in Istanbul Medical School, Department of General Surgery (Turkey) and in University of Catania Medical School, Department of Surgery (Italy), we performed laparoscopic splenectomy in three cases of splenic lymphangioma. RESULTS: These three female patients, with the age of 26, 30 and 40, had nonspecific abdominal pain requiring abdominal CT scan and magnetic resonance imaging, which showed incidental cystic lesions in the spleen, associated with cholelithiasis in one case. Preoperative laboratory tests and physical examinations were normal. Laparoscopic splenectomy was performed successfully with three 10 mm trocars in two patients in less than 1 hour, and with an Hasson trocar, two 5 mm trocars and one 10-12 mm trocar in the last case, who required simultaneous cholecystectomy. No peroperative and postoperative complications has occurred. Histopathological examinations confirmed the preoperative diagnosis. CONCLUSION: Laparoscopic splenectomy is the best treatment for patients with suspected cystic lymphangioma. It permits a total pathological examination of the spleen, and it should be preferred to partial splenectomy because of possible multiple lesions. In conclusion, minimal invasive treatment of this rare pathology is an effective and safe procedure. PMID- 17915060 TI - Stem cells: three questions. PMID- 17915062 TI - An international benchmarking collaboration: measuring outcomes for the hypertensive disorders of pregnancy. AB - OBJECTIVE: To implement a set of clinical indicators to benchmark outcomes for women suffering from the hypertensive disorders of pregnancy. METHODS: Seven clinical indicators were designed and applied retrospectively to data collected from two tertiary referral centres, Royal Prince Alfred Hospital, Sydney, Australia and British Columbia Women's Hospital and Health Centre, Vancouver BC, for all women coded as hypertensive during pregnancy under the International Classification of Disease (ICD-10) coding system in the years 2002-2004. Diagnostic categories were assigned using the Australasian Society for the Study of Hypertension in Pregnancy criteria, expressed in equivalent Canadian terms drawn from the Report of the Canadian Hypertension Society Consensus. Comparisons were made using the established clinical indicators. Data analysis using chi square comparison was performed with significance set at P < 0.05. Seven outcome measures of maternal and neonatal mortality and morbidity were compared. RESULTS: Significant areas of difference between the two tertiary referral centres were seen in birth weights below the 10th centile (RPA 11% vs. BCW 20%; P < 0.05) and below the 3rd centile (RPA 1.5% vs. BCW 7.5%; P < 0.001). There were significantly more episodes of maternal pulmonary edema at BCW than at RPA (0.1% and 1.2%, respectively; P < 0.001). CONCLUSION: Between similar centres, clinically significant differences in outcomes for HDP were identified. Further evaluation of differences may lead to analysis of possible contributors such as expectant versus urgent delivery management policies, rigidity of blood pressure control, and choice of antihypertensive drug. PMID- 17915061 TI - Dalteparin and low-dose aspirin in the prevention of adverse obstetric outcomes in women with inherited thrombophilia. AB - OBJECTIVE: To evaluate the benefit of treatment with dalteparin and low-dose aspirin (ASA) in the prevention of obstetric complications in women with inherited thrombophilia. METHODS: A retrospective chart review identified women who had had at least one pregnancy complicated by severe early-onset preeclampsia, placental abruption, fetal growth restriction (FGR), or fetal death. The following inherited thrombophilias were included: deficiencies of antithrombin, protein C, or protein S, and mutations of factor V Leiden (G1691A), factor II (G20210A), or methylenetetrahydrofolate reductase C677T. RESULTS: The records of 43 women with 110 pregnancies were included in the study. Anticoagulant prophylaxis was administered using dalteparin in 13 pregnancies, ASA with dalteparin in 26, and ASA alone in 11. Dalteparin alone and ASA alone showed equivalent effects in preventing preeclampsia and FGR. Combined dalteparin and ASA significantly decreased the risk of preeclampsia (odds ratio [OR] 0.80; 95% confidence intervals [CI] 0.70-0.91, P = 0.001) and FGR (OR 0.70; 95% CI 0.60 0.82, P = 0.001). CONCLUSION: Data from this retrospective cohort study suggest that combined treatment with dalteparin and ASA decreases the risk of preeclampsia by 20% and the risk of FGR by 30% in women with inherited thrombophilia. PMID- 17915063 TI - Obstetrician or family physician: are vaginal deliveries managed differently? AB - BACKGROUND: In Canada, obstetricians and family physicians both provide obstetrical care. However, the effect of specialty training on obstetrical outcomes of low-risk pregnancies has not recently been evaluated. In this study we examine the role of specialty training on the management of vaginal deliveries. METHODS: We conducted a cohort study on all vaginal deliveries that took place at Sacre-Coeur Hospital between July 2000 and June 2006. We compared baseline characteristics of obstetricians and family physicians and used an unconditional logistic regression model to estimate the adjusted relative risk of undergoing different obstetrical interventions. RESULTS: Of a total 8807 vaginal deliveries, 1915 were conducted by eight obstetricians and 6892 were conducted by 21 family physicians. Apart from a higher rate of induction of labour in patients of obstetricians, baseline characteristics were comparable between the two groups. Overall rates of use of instruments were similar in the two groups; however, family physicians were less likely than obstetricians to perform an episiotomy (odds ratio [OR] 0.47; 95% confidence intervals [CI] 0.41-0.55) but more likely to have patients who sustained a perineal injury (OR 1.51; 95% CI 1.36-1.68). There were no differences in the incidence of third- and fourth degree tears, and 5-minute Apgar scores were similar in both groups. CONCLUSION: Obstetricians and family physicians differ in the performance of episiotomies, and their patients differ in the resulting type of perineal injury. Instrument use and neonatal outcomes were similar in both groups. Major maternal and neonatal morbidity are unlikely to differ whether women with low-risk pregnancies are delivered by an obstetrician or a family physician. PMID- 17915064 TI - Social inequalities in use of prenatal care in Manitoba. AB - OBJECTIVE: Analysis of regional variations in use of prenatal care to identify individual-level and neighbourhood-level determinants of inadequate prenatal care among women giving birth in the province of Manitoba. METHODS: Data were obtained from Manitoba Health administrative databases and the 1996 Canadian Census. An index of prenatal care use was calculated for each singleton live birth from 1991 to 2000 (N = 149,291). Births were geocoded into 498 geographic districts, and a spatial analysis was conducted, consisting of data visualization, spatial clustering, and data modelling using Poisson regression. RESULTS: We found wide variation in rates of inadequate prenatal care across geographic areas, ranging from 1.1% to 21.5%. Higher rates of inadequate care were found in the inner-city of Winnipeg and in northern Manitoba. After adjusting for individual characteristics, the highest rates of inadequate prenatal care were among women living in neighbourhoods with the lowest average family income, the highest proportion of the population who were unemployed, the highest rates of recent immigrants, the highest percentage of the population reporting Aboriginal status, the highest percentage of single parent families, the highest percentage of the population with fewer than nine years of education, and the highest rates of women who smoked during pregnancy. CONCLUSION: Social inequalities exist in the use of prenatal care among Manitoba women, despite there being a universally funded health care system. Regional disparities in rates of inadequate prenatal care emphasize the need for further research to determine specific risk factors for inadequate prenatal care in socioeconomically disadvantaged neighbourhoods, followed by provision of effective targeted services. PMID- 17915065 TI - Self-collected samples for testing of oncogenic human papillomavirus: a systematic review. AB - BACKGROUND: To investigate the role of self-sampling for human papillomavirus (HPV) testing as an alternative to cervical cancer screening by clinicians (i.e., Papanicolaou [Pap] test). METHODS: A systematic search of MEDLINE, EMBASE, Cochrane Library, and other sources for evidence related to the efficacy and feasibility of HPV DNA self-collection. RESULTS: A total of 25 studies were identified. In 22 comparisons across 19 studies, the concordance between samples collected by patients and those obtained by clinicians was reasonably high in the majority of cases. Women in many countries across wide age ranges were successful in collecting samples for HPV DNA testing. In four studies, the quality of the cytology from patient samples was as good as clinician samples, with more than 95% of samples yielding HPV DNA results. The studies that examined acceptability found that women were generally very positive about collecting their own samples, although some concerns were noted. No study evaluated the effect of HPV DNA self sampling on screening participation rates, early detection, survival, or quality of life. CONCLUSIONS: Self-sampling for HPV DNA testing is a viable screening option, but there is insufficient evidence to conclude that self-sampling for HPV DNA testing is an alternative to the Pap test. Although HPV DNA testing using self-collected samples holds promise for use in under-resourced areas or for women who are reluctant to participate in Pap testing programs, the evidence supporting it is limited. Further definitive research is needed to provide a solid evidence base to inform the use of self-sampling for HPV DNA testing for the purpose of increasing screening rates, especially in women who are never or seldom screened. PMID- 17915066 TI - Acceptability of reminder letters for Papanicolaou tests: a survey of women from 23 Family Health Networks in Ontario. AB - OBJECTIVE: To explore women's perspectives on the acceptability and content of reminder letters from the family physician for Papanicolaou (Pap) test screening and the effect of reminder letters on compliance with screening recommendations. METHODS: A population-based survey was conducted in 23 Family Health Networks and Primary Care Networks participating in a demonstration project to increase the delivery of preventive services in Ontario. Questionnaires were mailed to randomly selected women aged 35 to 69 years who had received a reminder letter for a Pap test from their family physician within the previous six months. Two focus groups were conducted with a volunteer sample of respondents. RESULTS: The usable response rate was 54.3% (406/748). Two-thirds (65.8%, 267/406) of women who completed the survey recalled receiving the reminder letter. Overall, 52.3% (212/405) reported having a Pap test in the past six months. Among women who recalled the reminder letter and scheduled or had a Pap test, 71.4% (125/175) reported that the letter influenced their decision to be screened. The majority of respondents (80.8%, 328/406) wanted to continue to receive reminder letters for Pap tests from their physician, and 34.5% (140/406) wanted to receive additional information about cervical screening. Focus group interviews indicated that women who have had a Pap test may still be unsure about screening recommendations, what the test detects, and the rationale for follow-up procedures. CONCLUSIONS: Reminder letters in family practice were viewed as useful and influenced women's decisions to undergo Pap test screening. Women who have had a Pap test may still need additional information about the test. PMID- 17915067 TI - Maternal luteoma of pregnancy presenting with virilization of the female infant. AB - BACKGROUND: Virilization in female newborns typically results from congenital adrenal hyperplasia, requiring immediate diagnosis and treatment. We report a rare cause of virilization, maternal pregnancy luteoma, responsible for virilization of both a newborn and the mother. Luteomas are usually asymptomatic tumour-like ovarian lesions of pregnancy that secrete androgens in only 25% of cases. Many female infants born to masculinized mothers will also be virilized. CASE: A term infant born with ambiguous genitalia was transferred to a referral centre for investigation, diagnosis, and treatment. Assessment identified Prader II-III genitalia, an elevated serum testosterone level, a normal serum 17 hydroxyprogesterone level, and a normal female karyotype (46,XX). The mother had had virilization from the second trimester and was found to have an elevated serum testosterone level. Pelvic ultrasound assessment in the mother showed a complex right ovarian mass. Laparotomy was performed, and the mass was excised. Histopathology examination confirmed a luteoma. CONCLUSION: High maternal serum testosterone levels due to a luteoma can result in virilization in the female newborn. This report emphasizes the need to consider possible underlying maternal pathology in evaluating a virilized female infant. PMID- 17915068 TI - When it comes to pregnant women sleeping, is left right? AB - Pregnant women who lie in a supine position may develop syncopal symptoms. However, of those women who become symptomatic, only 2% to 4% have significant aortocaval compression. Even in this small minority of symptomatic women, there is no evidence of fetal compromise. The advice often given to pregnant women to lie on the left side is therefore not relevant. In some women, experiencing a pre syncopal episode will cause them to avoid lying in the supine position. PMID- 17915069 TI - Stem cell research ethics: consensus statement on emerging issues. AB - This article is a consensus statement by an international interdisciplinary group of academic experts and Canadian policy-makers on emerging ethical, legal and social issues in human embryonic stem cells (hESC) research in Canada. The process of researching consensus included consultations with key stakeholders in hESC research (regulations, stem cell researchers, and research ethics experts), preparation and distribution of background papers, and an international workshop held in Montreal in February 2007 to discuss the papers and debate recommendations. The recommendations provided in the consensus statement focus on issues of immediate relevance to Canadian policy-makers, including informed consent to hESC research, the use of fresh embryos in research, management of conflicts of interest, and the relevance of public opinion research to policy making. PMID- 17915070 TI - A new medication for the worst addiction. PMID- 17915071 TI - A second varenicline trial. PMID- 17915072 TI - Varenicline as maintenance therapy. PMID- 17915074 TI - Why buprenorphine is so successful in treating opiate addiction in France. AB - In France, all registered medical doctors have been allowed to prescribe buprenorphine without any special education or licensing since 1995. This has led to a rapidly increasing number of opiate-dependent users under buprenorphine treatment in primary care. French physician compensation mechanisms, pharmacy services, and medical insurance funding all have contributed to minimizing barriers to buprenorphine treatment. Approximately 20% of all physicians in France are prescribing buprenorphine to treat more than one half of the estimated 180,000 problem heroin users. Intravenous diversion of buprenorphine may occur in up to 20% of buprenorphine patients and has led to relatively rare overdoses in combination with sedatives, whereas total opiate overdose deaths have declined substantially. In France, buprenorphine maintenance treatment for problem opiate users was feasible and safe through office-based prescriptions in a relaxed regulatory environment. PMID- 17915073 TI - Genetics and smoking behavior. AB - Accumulating data support the role of genetic factors in smoking initiation, progression to tobacco dependence, and smoking persistence. This review summarizes current research on the heritability of tobacco use phenotypes and genetic association studies of smoking-related behaviors. Although progress has been made in genetics research on smoking behavior, many studies have methodological limitations, including insufficient samples for detecting gene gene and gene-environment interactions and use of less refined phenotypes. Pharmacogenetic investigations also are identifying variants in drug-metabolizing enzymes, receptors, and transporters that modify therapeutic response to smoking cessation medications; however, the field is relatively new, and most findings in this area have yet to be replicated. As this research advances, it will be important to study and address practical, economic, ethical, and social barriers to the translation of genetics research on tobacco use to clinical practice. PMID- 17915075 TI - Targeted modulators of the endogenous cannabinoid system: future medications to treat addiction disorders and obesity. AB - The endogenous endocannabinoid system encompasses a family of natural signaling lipids ("endocannabinoids") functionally related to (9)-tetrahydrocannabinol, the psychoactive ingredient of marijuana (cannabis), along with proteins that modulate the endocannabinoids, including enzymes, transporters, and receptors. The endocannabinoid system's ubiquitous regulatory actions in health and disease underscore its importance to mammalian (patho)physiology and suggest discrete targets through which it may be modulated for therapeutic gain. Medications based on the endocannabinoid system are an important focus of contemporary translational research, particularly with respect to substance abuse and obesity, two prevalent disorders with a pathogenic component of endocannabinoid system hyperactivity. Pressing health care needs have made the rational design of targeted CB1 cannabinoid-receptor modulators a promising route to future medications with significant therapeutic impact against psychobehavioral and metabolic disturbances having a reward-supported appetitive component. PMID- 17915076 TI - Treatment of patients comorbid for addiction and other psychiatric disorders. AB - Psychiatric disorders and drug and alcohol use disorders commonly co-occur. A growing literature has documented the epidemiology and effects on the course of illness of comorbid psychiatric and substance use disorders (SUDs). Advances in treatment of co-occurring illnesses have progressed more slowly. The current article reviews recent developments in the diagnosis and treatment of co occurring psychiatric disorders and SUDs with particular focus on psychotic disorders, affective disorders, anxiety disorders, personality disorders, and attention-deficit/hyperactivity disorder. Current treatment options and implications for future research are highlighted. PMID- 17915077 TI - The future of vaccines in the management of addictive disorders. AB - Conventional substance abuse treatments have had only limited success. As a result, new approaches, including vaccination to block the effects of drugs such as cocaine, nicotine, methamphetamine, and phencyclidine, are in development. Although a number of possible rationales for the effects of antidrug vaccines have been suggested, the most straightforward and intuitive mechanism would involve binding of the drug by antibodies in the bloodstream, thereby blocking entry or reducing the rate of entry of the drug into the central nervous system. The theoretical parameters that would influence vaccine-induced drug pharmacodynamics are presented in this review, along with the current status on vaccine development for nicotine, cocaine, methamphetamine, and phencyclidine. PMID- 17915079 TI - Sleep disturbances and ADHD medications. PMID- 17915078 TI - The role of stress in addiction relapse. AB - Stress is an important factor known to increase alcohol and drug relapse risk. This paper examines the stress-related processes that influence addiction relapse. First, individual patient vignettes of stress- and cue-related situations that increase drug seeking and relapse susceptibility are presented. Next, empirical findings from human laboratory and brain-imaging studies that are consistent with clinical observations and support the specific role of stress processes in the drug-craving state are reviewed. Recent findings on differences in stress responsivity in addicted versus matched community social drinkers are reviewed to demonstrate alterations in stress pathways that could explain the significant contribution of stress-related mechanisms on craving and relapse susceptibility. Finally, significant implications of these findings for clinical practice are discussed, with a specific focus on the development of novel interventions that target stress processes and drug craving to improve addiction relapse outcomes. PMID- 17915080 TI - Recent advances in structural and functional brain imaging studies of attention deficit/hyperactivity disorder. AB - The field of neuroimaging of attention-deficit/hyperactivity disorder (ADHD) is now 30 years old. This brief selective review highlights the increasing sophistication of recent structural and functional neuroimaging studies of ADHD. In volumetric studies, investigators are examining extra-frontal, as well as frontal-striatal circuits and beginning to differentiate the potential effects of medication exposure. Functional MRI studies are focusing on familial/genetic influences and enrolling medication naive, as well as medicated children with ADHD. A promising trend is the application of resting state approaches to mapping functional connectivity, which provides unexpectedly detailed information about interregional relationships while bypassing potentially confounding issues related to task performance. These developments allow us to conclude that neuroimaging studies of ADHD will increasingly inform our understanding of the neuronal substrates of ADHD. PMID- 17915081 TI - When ADHD and substance use disorders intersect: relationship and treatment implications. AB - There has been increasing interest in the overlap between attention deficit/hyperactivity disorder (ADHD) and substance use disorders (SUDs). In this report, we describe the developmental relationship between ADHD and SUDs. ADHD alone and in combination with co-occurring psychopathology is a risk factor for the development of SUDs in adulthood. Conversely, approximately one fifth of adults with SUDs have ADHD. Pharmacotherapeutic treatment of ADHD in children reduces the risk for later cigarette smoking and SUDs in adulthood. In contrast, medication treatment alone of adults with ADHD and current SUD is inadequate for both ADHD and SUD. Stimulant diversion continues to be of concern, particularly in older adolescents and young adults. PMID- 17915082 TI - Concurrent ADHD and bipolar disorder. AB - Attention-deficit/hyperactivity disorder (ADHD) frequently is present concurrently with bipolar disorder (BPD) in youth. This concurrence appears to be more common in younger children. The degree to which ADHD is present in adults with BPD has not been well studied. The psychiatric and behavioral symptoms associated with ADHD and BPD have significant overlap. The core symptoms of BPD are relatively independent and respond to different pharmacologic and behavioral strategies. Although much symptomatic overlap exists between ADHD and BPD, these conditions can be reliably differentiated from each other and require independent treatments that frequently need to be sequenced. PMID- 17915083 TI - Clinical recommendations in current practice guidelines for diagnosis and treatment of ADHD in adults. AB - Attention-deficit/hyperactivity disorder (ADHD) is a lifelong neurodevelopmental disorder in which approximately two thirds of patients experience impairment in adulthood. Although some adults with ADHD were diagnosed as children, many are first diagnosed as adults. This poses particular challenges given the limited familiarity with ADHD of many adult mental health services. As a result, several organizations, including the Canadian ADHD Resource Alliance, the American Academy of Child and Adolescent Psychiatry, the National Institutes of Health, and the British Association for Psychopharmacology, have developed practice guidelines for the assessment and treatment of adults with ADHD. This article reviews those guidelines in order to examine current best practices in adult ADHD. There is considerable agreement among these guidelines, which should be a critical part of moving from emerging knowledge to patient care, although both empirical evaluation and ongoing updates as new knowledge emerges will be important for their future development. PMID- 17915084 TI - Current status of cognitive-behavioral therapy as a psychosocial treatment for adult attention-deficit/hyperactivity disorder. AB - A convergence of research has established that attention-deficit/hyperactivity disorder (ADHD) is a valid clinical syndrome affecting individuals of all ages. ADHD is associated with significant impairment in many important life domains that often requires clinical intervention. Although medications are a well researched and effective first-line treatment option, many adult patients with ADHD continue to experience significant functional impairment despite the symptom improvement afforded by pharmacotherapy. Consequently, adjunctive psychosocial treatments often are indicated for adults with ADHD. The aim of this paper is to review the status of cognitive-behavioral therapy (CBT) as an effective psychosocial treatment (when combined with pharmacotherapy) for adult ADHD. This review includes a brief discussion of the CBT conceptualization of ADHD, a summary of clinical outcome studies of psychosocial treatments for adult ADHD, directions for future research, and a discussion of the various possible mechanisms of change involved in CBT for adult ADHD. PMID- 17915085 TI - ADHD genetics: 2007 update. AB - Attention-deficit/hyperactivity disorder (ADHD) is highly heritable. Confirmed association has been reported for several candidate genes, including DAT1, DRD4, SNAP-25, DRD5, 5HTT, HTR1B, and DBH; however, these confer relatively small risk. Family-based linkage studies have identified a number of chromosomal regions containing potential ADHD predisposing loci, some overlapping in two or more studies, including 5p, 6q, 7p, 11q, 12q, and 17p. New large-scale studies that apply recent technological advances to perform high-density genotyping of the entire genome, in combination with information on the haplotype structure of the human genome, now allow testing of single-nucleotide polymorphism association with disease phenotype without any a priori hypothesis. They may contribute further to our understanding of the genetic factors involved in ADHD. The heterogeneous complex ADHD phenotype, as well as epigenetic factors may be contributing to the challenge of genetic studies. Samples that include limited age ranges may have better success at uncovering genes whose expression is limited to specific developmental stages. PMID- 17915086 TI - Macrophage migration inhibitory factor and its genetic association with arthritis: a work in progress. PMID- 17915087 TI - Sulfasalazine in undifferentiated spondyloarthropathies. PMID- 17915088 TI - Is low-dose infliximab every 8 weeks effective in treating ankylosing spondylitis? PMID- 17915089 TI - Another look at low-dose infliximab in treating ankylosing spondylitis. PMID- 17915090 TI - Advances in musculoskeletal imaging and their clinical utility in the early diagnosis of spondyloarthritis. AB - Interest in imaging ankylosing spondylitis (AS) and related spondyloarthropathies has increased in recent years. MRI is regarded as the most sensitive imaging modality to detect early inflammatory lesions in the sacroiliac joints and in the spine. Standard radiography plays a major role in evaluating potential disease modifying properties of the recently introduced, symptomatically effective anti tumor necrosis factor-alpha agents. Various radiographic scoring methods have been developed to assess and monitor spinal structural damage in AS. The modified stoke ankylosing spondylitis spine score is the method of choice based on its sensitivity to change. MRI is emerging as a useful tool to detect AS early, but this remains a challenge. Recent progress in MR technology with the advent of whole body MRI has expanded the potential role of imaging in AS. Further data are needed before MRI can replace radiographs for earlier diagnosis of AS. PMID- 17915091 TI - Undifferentiated spondyloarthritis: a global perspective. AB - This paper reviews the concept and outcome of undifferentiated spondyloarthritis (SpA) as reported in retrospective and prospective studies from different parts of the world. Although the designs and definitions vary across different studies, clearly a significant proportion of patients with undifferentiated SpA will fulfill ankylosing spondylitis within 10 years in association with certain prognostic factors. The paper also examines the concept of axial SpA and its importance as an early indicator of ankylosing spondylitis. Ultimately, the recognition of undifferentiated SpA and axial SpA may lead to early treatment with highly efficacious drugs. PMID- 17915092 TI - Diagnosing early ankylosing spondylitis. AB - Diagnosis of ankylosing spondylitis is still delayed by many years. Several efforts have been made in the past few years to shorten this delay. A new set of criteria for inflammatory back pain has performed better than previous sets. MRI has evolved to become the standard imaging modality for the detection of sacroiliitis during early disease, and it clearly outperforms quantitative scintigraphy, which was the standard screening test for many years. Promising new developments such as whole body MRI and ultrasound (sonography) for the detection of enthesitis or sacroiliitis deserve further evaluation. Serum antibodies directed against a 28-kD Drosophila antigen may provide additional diagnostic information. A recently proposed diagnostic algorithm in patients with suspected early ankylosing spondylitis may help physicians confidently diagnose patients before definite radiographic sacroiliitis is detectable. Finally, referral strategies for patients seen by primary care physicians seem to work well and are currently under further valuation. PMID- 17915093 TI - Appropriate management of axial spondyloarthritis. AB - The management of axial spondyloarthritis includes a structured baseline assessment of the disease and follow-up of treatment efficacy using validated instruments. The treatment will depend on the severity and predominant manifestations of the disease. The cornerstone of management remains physical therapy and nonsteroidal anti-inflammatory drugs. Disease-modifying antirheumatic drugs such as sulfasalazine have shown efficacy only in treating peripheral arthritis, whereas thalidomide and pamidronate have shown some efficacy in treating axial inflammation. Anti-tumor necrosis factor-alpha agents are an efficacious option for treating signs and symptoms of axial disease, peripheral arthritis, and enthesitis, and for improving functional outcomes. They have not shown efficacy in reducing radiographic progression in axial disease. Recent evidence suggests that the new bone formation in ankylosing spondylitis may be due to upregulation of Wnt signaling in the osteoblastic pathway secondary to low serum DKK-1 levels, which are further suppressed by tumor necrosis factor inhibitor therapy. PMID- 17915094 TI - Genetics of ankylosing spondylitis: an update. AB - Substantive evidence exists that genetic factors play a pivotal role in susceptibility to ankylosing spondylitis (AS). HLA-B27 remains the most convincing and universal association of a genetic factor with AS. Over the last decade there has been immense interest in elucidating genetic variants outside the major histocompatibility complex region. Due to larger AS datasets along with recent advancements in the characterization of genetic markers and large-scale genotyping platforms, replicated non-major histocompatibility candidates have now emerged. This article reviews the current evidence regarding the genetics of AS, with an emphasis on the recent major advances, and it discusses the challenges and limitations in interpreting these studies. PMID- 17915096 TI - B-cell inhibitors as therapy for rheumatoid arthritis: an update. AB - A revolution in the treatment of rheumatoid arthritis has occurred in recent years. This holds particularly true for B-cell-directed therapies for rheumatoid arthritis. The approval of rituximab for the treatment of rheumatoid arthritis has not only expanded the armamentarium of therapies for rheumatologists, but it has also led the way to better understanding of the biologic sequelae of these treatments as well as the potential to better understand the etiology of autoimmune diseases. This review updates the latest B-cell therapies in rheumatoid arthritis. PMID- 17915095 TI - Antibodies to citrullinated proteins: pathogenic and diagnostic significance. AB - The recent demonstration of efficacy in the treatment of rheumatoid arthritis with B-cell-depleting therapy, along with the increasing understanding of the role of anticitrullinated protein antibodies (ACPA) have begun to provide new insights into the pathogenesis of this complex disease. This manuscript reviews the current understanding of ACPA with regard to clinical associations and pathogenic mechanisms in preclinical animal models. These data are synthesized into a model of the development of rheumatoid arthritis that refocuses attention on the role of B cells and immune complexes, returning in part to the origins of scientific investigation in this disease, and points to directions of research that are necessary to achieve the full therapeutic, diagnostic, and prognostic potential of the ACPA antigen-antibody system. PMID- 17915097 TI - Vaccine responses in patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a systemic autoimmune disease that is associated with immunologic alterations in T cells and B cells. Moreover, many of the agents used in RA patients are potentially immunosuppressive. Thus, the underlying disease and treatment may both increase the susceptibility to infections and decrease vaccine responses. With the growing use of aggressive therapies for RA, including anti-tumor necrosis factor agents and newer biologic therapies such as rituximab and abatacept, an increasing concern will be that patients may not respond to conventional vaccination. Further prospective studies on response to vaccination are needed to answer this important public health question. Nevertheless, it is already clear that vaccination does induce response in many patients. Unfortunately, vaccination is underutilized in RA patients and needs to be aggressively promoted. PMID- 17915098 TI - Switching anti-TNF-alpha agents: what is the evidence? AB - The availability of biologic agents targeting tumor necrosis factor (TNF)-alpha represents a significant advance in the management of rheumatoid arthritis. Anti TNF-alpha therapy has been associated with dramatic improvements in the clinical signs and symptoms of rheumatoid arthritis and has been shown to greatly retard the destructive process that too often characterizes this condition. Although effective and well-tolerated in a substantial proportion of patients, primary and secondary failures of anti-TNF-alpha strategies have been well described, affecting up to one-third to one-half of subjects treated with these agents. Switching from one anti-TNF-alpha agent to a second (or even third) anti-TNF alpha therapy has emerged as a means of addressing treatment failures with this drug class. This review examines data addressing the practice of switching anti TNF-alpha agents in the context of initial treatment failure, with a focus on data from peer-reviewed reports. PMID- 17915100 TI - [School performance--no difference between twins and singletons--secondary publication]. AB - Studies of twins born 50+ years ago indicate that twins had a lower IQ than singletons. We carried out a register based study of all twins (N=3411) and a 5% random sample of singletons (N=7796) born in Denmark 1986-1988. We found no difference in school performance in adolescence between twins and singletons, nor when controlling for birth weight, gestational weight, age of parents or parents' education. Progress within living conditions, obstetrics, and paediatrics may be the reason why twins now generally have school performances similarly to singletons. PMID- 17915099 TI - Malignancy and rheumatoid arthritis. AB - The occurrence of cancer is not an infrequent event in patients with rheumatoid arthritis (RA). Indeed, following diagnosis of RA at a typical age (55 years), one in five patients will be diagnosed with cancer. In the vast majority of such cases, the cancer has nothing to do with RA or its treatment; rather, it represents the "background" risk applicable to all humans. In some cases, the cancer occurs as a result of factors also associated with the risk of developing RA (eg, smoking), even though no direct link exists between the cancer and the RA. In a fraction of cases, however, the cancer is causally associated with the RA disease or its treatments. This review summarizes our current understanding of the occurrence of cancer in RA, possible links to RA disease and to traditional and newer RA treatments. PMID- 17915102 TI - [What is important for the family of patients in the intensive care unit?]. AB - The abundant bibliography shows us that the family plays a fundamental paper in the recovery of the hospitalized patients, and the patients in the intensive care units (ICU) are no exception. Several authors have demonstrated that a stress free and calm setting helps in the care process. Thus, the family needs must be considered to improve their well-being. In this way, their attitude will help in the patient's recovery. Health care personnel must be aware of these needs and make up for their deficit as far as possible. The first study that evaluated this question used the CCFNI questionnaire (Critical Care Family Needs Inventory) that measures the degree of importance given by the relatives to the needs of safety, support, information, comfort and proximity. This study has aimed to detect the degree of importance given by the relatives of the patients in the ICU of the Hospital Dr. Josep Trueta of Girona, using the CCFNI questionnaire. All the relatives of the patients in said unit during the autumn of the year 2003 were interviewed and the needs considered to be most important were compared with the sociodemographic variables of the relatives. The total sample was made up of the 132 people interviewed. The need considered to be most important was safety with 50.3% followed by need for support with 47.2%. In the results, some differences were observed depending according to age and studies of those surveyed and according to the diagnosis and the origin of the hospitalization. The collected data did not differ with the reviewed bibliography. All the study was very valuable for the health care professionals of the unit and was taken into account to improve the deficiencies of the ICU of the hospital of Girona. PMID- 17915103 TI - [Proposal of a new assessment scale of work load and nursing times (VACTE]. AB - The scale Nine Equivalents of nursing Manpower use Score (NEMS) for the evaluation of the nursing care loads is the most well known and applied worldwide. Nevertheless, we have found a series of limitations: it does not reflect the "proper nursing activity" but only the cares related to the medical intervention. Furthermore, it is directly related to severity while integral attention to the patient implies an infinity of cares, which are not necessarily related to the severity. In addition, we understand that the planned personnel ratios may be unsuitable, with the consequent repercussions for the patient, nurses and the sanitary institution. The primary targets were: elaboration of a representative scale of all the cares and tasks made by the nurses (VACTE) in our unit, to determine if it is more precise and objective than NEMS for the measurement of the service loads and to calculate the operative ratio patient nurse based on the new proposed scale. We made a descriptive and retrospective study on 91 patients admitted to the Intermediate Care Unit of the Fundacion Hospital de Jove during the first three months of 2004. Previously we created scale VACTE, making real measurements of the time inverted in the execution of each one of the cares in 50 patients. Later, a comparison was made between the APACHE II, NEMS and VACTE scales, taking as reference the scores obtained in the same ones during the first 24 hours of the stay. The statistical analysis was made by SPSS 11.0, assuming a confidence level of 95% (p < 0.05): lineal analysis of simple regression to compare the different scales; the force of its correlation with Spearman's coefficient and we compared the independent dichotomize variables with the Mann-Whitney test. The main results determined after the study were the following: regarding the scale to evaluate seriousness applied to the patients, an average APACHE II score of 12.1 +/- 5.9 was obtained. The average value with the NEMS was 19.5 +/- 5.7 and the average one with VACTE was 365 +/- 91.2. Significant differences were found between APACHE II and the NEMS (p = 0.008) and VACTE (p < 0.001) scales. This relationship presented more force with VACTE (r = 0.43; p < 0.001) than with NEMS (r = 0.23; p = 0.002). The operative patient-nurse ratio calculated with VACTE was 2.01. Scale VACTE is representative of nursing activity and seems to be more effective than NEMS to determine service loads. The ratio planned in the unit duplicated the calculated operative ratio. PMID- 17915104 TI - [Nursing in Intensive and Coronary Care. Bibliometric analysis of 180 original articles]. AB - INTRODUCTION: Analyze the "original" type articles published in Spain within the field of Intensive and Coronary Care Nursing with bibliometric methodology. MATERIAL AND METHODS: A total of 180 articles classified by different journals as original were analyzed. The following information was collected: year of publication, journal, number of authors per article, analysis of thematic by subject and sub-theme, statistical procedure used and references as well as existence or not of institutional collaboration and territorial origin of the articles. RESULTS: Compliance of Price Law, obtaining duplication rate of 2.16 years. The Journal Enfermeria Intensiva is the Nucleus of Bradford. The mean number of authors per article is 4.82. The type of original used most is cross sectional cut (47.22% of the total), followed by qualitative type (22.78%). The subject matter studied most was psychology (23.3%), followed by respiratory (15.6%). The statistics used most was descriptive (percentage, median and standard deviation), followed by inferential (parametric test, 67.79% and non parameter test in 23.31%). Crude rate of references to journals was 12.93, and to books, 3.06. In English, it was 10.94, and in Spanish, 7.53. A total of 73.9% of the articles were written without any collaboration and in 19.44% of them, there was collaboration between the hospital site and university. The most product hospital site was the Clinica Universitaria de Pamplona (7.22% of the total), the University Nursing School that of the University of Navarra (10 articles), the province, Barcelona (25.14%), and the Regional Community that of Catalonia, with 26.23% of the total. CONCLUSIONS: Intensive and Coronary Care Nursing is a young discipline with a growth model that complies with the Price Law and rapid growth rhythm. The Nucleus of Bradford of the journals is made of by Enfermeria Intensiva. The mean number of authors per article is elevated in comparison with other fields of Nursing, although less than foreign nursing studies. There is a significant deficit in the number of crude references per article used, with predominance of the English language and journals. The limited existence of institutional collaboration makes us consider a still non-established discipline, although with an ascending clear tendency in recent years. PMID- 17915105 TI - [Delirium in intensive care. Incidence and risk factors]. AB - Delirium has been documented as a common and serious problem in hospital settings, but its recognition in critically ill patients is made difficult by the inability to interview intubated patients and by the presence of drugs. The objectives of this study were to describe the incidence of delirium in the Intensive Care Unit (ICU) and to determine some of the risk factors. It is an analytical, observational and prospective study in an 8-bed general ICU. The participants were 112 consecutive patients aged 18 and older admitted to the ICU for three or more days. Drug consumers and patients with psychiatric disease or cerebral disease were excluded. Daily ratings of the Intensive Care Delirium Screening Checklist (ICDSC) were made by the staff nurses. Other data such as patient's diseases, drug prescriptions and APACHE II were collected. Delirium was present in 11 out of 100 patients-day. APACHE II > or = 15, sedative drugs and mechanical ventilation were risk factors for delirium in ICU. The conclusions of this study have determined that delirium is a frequent complication in ICU and that severity of illness, sedative drugs and being intubated are risk factors. Moreover, screening for delirium in the ICU with a validated scale can be made by the staff nurses. PMID- 17915106 TI - HIV/AIDS burden in rural Africa: the people's struggle and response of the international community. PMID- 17915107 TI - [Enterococcus: an emerging pathogen in our hospitals?]. PMID- 17915108 TI - [An analysis of 182 enterococcal bloodstream infections: epidemiology, microbiology, and outcome]. AB - INTRODUCTION: Enterococcal bloodstream infections have acquired considerable importance in recent years, mainly because of the increasing number of cases that occur during hospital admission. METHODS: Retrospective study of the clinical records of patients diagnosed with enterococcal bacteremia and hospitalized over a 12-year period (January 1994-April 2006), analyzing epidemiological, clinical and microbiological characteristics, outcome and prognostic factors. RESULTS: A total of 182 episodes of bacteremia were recorded; 68% of them were nosocomial infections, accounting for 5% of the in-hospital bacteremia episodes in this period. The most frequent sources of infection were urinary tract (29%), cardiovascular (25%), intra-abdominal (21%) and primary bacteremia (12%). Associated comorbid conditions were present in 85% of patients, mainly neoplasms (33%). Enterococcus faecalis was responsible for 70% of cases, E. faecium 22%, and other species of enterococci 8%. Twenty percent were polymicrobial bacteremia. Antibiotic resistance was documented in 23% of the strains: 14% ampicillin, 8% gentamicin, 3% ampicillin and gentamicin, and 0.5% vancomycin. Overall mortality was 31%. Polymicrobial bacteremia and comorbidity were associated with a poor prognosis. CONCLUSION: In our hospital, Enterococcus is the fifth most frequent cause of nosocomial bacteremia. E. faecium is characterized by a high incidence (more than 50% of cases) of ampicillin resistance. PMID- 17915109 TI - [Environmental contamination during a vancomycin-resistant Enterococci outbreak at a hospital in Argentina]. AB - INTRODUCTION: Vancomycin-resistant enterococci isolates (VRE) have caused numerous outbreaks in intensive care units (ICUs). A contaminated hospital environment, the hands of health care workers (HCW), and carrier patients may play important roles in perpetuating the chain of transmission in these outbreaks. The aims of this study were to report the first VRE outbreak in our center and assess the role of environmental contamination and HCW hands in the spread of new cases of enterococcal infection. MATERIAL AND METHOD: Between August and December 2003, surveillance cultures were performed with samples from all patients (n = 113) admitted to the ICU, as well as cultures of samples from the environment (n = 69) and HCW hands (n = 23). RESULTS: Eighteen clinical samples from 8 patients and 7 environmental samples yielded Enterococcus faecium (24 strains) and E. avium (1 strain). VRE was not detected on HCW hands. All the VRE isolates belonged to a single clone and carried the vanA gene. CONCLUSION: Environmental contamination provides an important reservoir for future outbreaks of VRE, perpetuating transmission of the microorganism in the hospital setting. PMID- 17915110 TI - [High-risk clonal complexes CC2 and CC9 are widely distributed among Enterococcus faecalis hospital isolates recovered in Spain]. AB - INTRODUCTION: Our previously described multilocus sequence typing (MLST) scheme for Enterococcus faecalis has provided insight into the population structure and global epidemiology of this organism. Two high-risk complexes, CC2 and CC9, especially adapted to the hospital environment and widely distributed in Europe and America, were identified. The purpose of this study was to define the presence of CC2 and CC9 among E. faecalis strains isolated in Spain. METHODS: A total of 81 E. faecalis isolates recovered from several sources and geographic areas of Spain were characterized using MLST. Because of their clinical and epidemiological interest, strains were included from each of the vancomycin resistant E. faecalis hospital outbreaks described in Spain. RESULTS: Among the isolates, CC2 and CC9 were detected in the hospital setting. Included in these CC were the vancomycin-resistant E. faecalis isolates causing hospital outbreaks in La Coruna, Palma de Mallorca and Valencia, as well as vancomycin-susceptible hospital isolates. The Index of Association (Ia), which measures linkage disequilibrium between alleles, revealed an epidemic population structure on a background of high recombination rates. CONCLUSIONS: High-risk complexes (CC2 and CC9) particularly adapted to the hospital environment were detected in Spain. Evolution of these CC in different areas depended on the local gene pool. Future infection control policies should be orientated to detect high-risk CC with the aim of predicting potential trends toward acquisition of specific resistance, such as to vancomycin. PMID- 17915111 TI - [Clinical practice guidelines from the Andalusian Society of Infectious Diseases (SAEI) for the treatment of tuberculosis]. AB - The therapeutic scheme for initial pulmonary tuberculosis recommended by the SAEI is as follows: Initial phase, isoniazid, rifampin and pyrazinamide given daily for 2 months. In HIV(+) patients and immigrants from areas with a rate of primary resistance to isoniazid > 4%, ethambutol should be added until susceptibility studies are available. Second phase (continuation phase): rifampin and isoniazid, given daily or intermittently for 4 months in the general population. HIV(+) patients (< or = 200 CD4) and culture-positive patients after 2 months of treatment should receive a 7-month continuation phase. A 6-month regimen is recommended for extrapulmonary tuberculosis, with the exception of tuberculous meningitis, which should be treated for a minimum of 12 months and bone/joint tuberculosis, treated for a minimum of 9 months. Treatment regimens for multidrug resistant tuberculosis are based on expert opinion. These would include a combination of still-useful first-line drugs, injectable agents, and alternative agents, such as quinolones. Patients who present a special risk of transmitting the disease or of non-adherence should be treated with directly observed therapy. PMID- 17915112 TI - [Effects of viral infection on transplant recipients]. AB - Viral infection remains an important cause of morbidity and mortality in transplant recipients. The risk of viral infection in these patients depends on several factors, such as the type of organ transplanted, the intensity of immunosuppression, and the recipient's susceptibility. In additional to direct effects, viral infection cause indirect effects, including greater risk of replication of other viruses, graft rejection, opportunistic infections and other specific entities for each type of transplant. These indirect effects result from the immunomodulatory activity of some viruses, such as cytomegalovirus and human herpes virus-6. For the most part, quantitative molecular tests have replaced serologic testing and in vitro culture for diagnosing infection. This approach is particularly prominent for cytomegalovirus, Epstein-Barr virus, hepatitis B virus, and hepatitis C virus. Despite these diagnostic advances, the development of specific antiviral agents and effective antiviral vaccines is limited. Thus, prophylactic strategies are still essential in transplant recipients. PMID- 17915113 TI - [Fungal endocarditis in a patient bearing a valve prosthesis]. PMID- 17915114 TI - [Fever, right hypochondrium pain and a hepatic mass with microcalcifications in a consumer of non-pasteurized dairy products]. PMID- 17915115 TI - [Neisseria mucosa endocarditis in a native valve]. PMID- 17915116 TI - Infection of a total hip arthroplasty due to Gemella morbillorum. PMID- 17915117 TI - [For whom the bell tolls]. PMID- 17915118 TI - From autonomy to accountability: individual versus system issues in contemporary practice. PMID- 17915119 TI - [Randomized clinical trial on the effectiveness of a postal reminder to increase tetanus-diphtheria vaccination coverage in the young adult population]. AB - OBJECTIVE: To evaluate the effectiveness of individual postal reminders compared with no action of any kind in increasing the tetanus-diphtheria immunization rate in the population aged between 24 and 30 years old in an area where a general population strategy was simultaneously being conducted through information posters on tetanus vaccination. METHOD: We performed an open, randomized clinical trial with parallel groups among the population aged 24-30 years old in a basic health area with 13,523 inhabitants over a 3-month period in 2005. There were 311 patients in the control group and 311 in the experimental group, after exclusion of individuals correctly vaccinated according to the computerized medical records system. The intervention evaluated was a postal reminder on tetanus vaccination. The response variable was the immunization rate due to the intervention. Other variables studied were sex, age, reason for seeking vaccination, compliance with the vaccination timetable, whether vaccination was rejected, and whether prior vaccination was verified. The statistical analysis consisted of Student's t-test and the chi2 test, with a confidence level of 95% (p < 0.05). RESULTS: Vaccination coverage among the general population at the end of 2005 was 75.6%. After the postal reminder, 22.2% of the control group and 40.5% of the experimental group were correctly vaccinated. CONCLUSIONS: Individual postal reminders sent to the population aged between 24 and 30 years is effective in increasing the immunization rate in this age group. Periodic reminders could help to ensure continuity in vaccination of the adult population. PMID- 17915120 TI - [Evaluation of the degree of implementation of tobacco control interventions in the Catalan Network of Smoke-Free Hospitals]. AB - OBJECTIVES: To evaluate the degree of implementation of tobacco prevention and control interventions and to determine the prevalence of smoking in the Catalan Network of Smoke-Free Hospitals according to the stage of adhesion to the project. METHOD: We performed an observational, descriptive, cross sectional study in 16 hospitals actively involved in the Catalan Network of Smoke-Free Hospitals (April 2005). Data were collected using the results of the self-audit, 9-item questionnaire of the European Network for Smoke Free Hospitals. To study tobacco consumption, we interviewed 3,896 employees. Data on sociodemographic characteristics, tobacco consumption, job category, tobacco control indicators, the project's stage of implementation, and adherence to the project were gathered. RESULTS: The prevalence of smoking was 35%. The implementation stage among all the hospitals studied was 53.3%. The items with high implementation scores (above 70%) were commitment, communication, consumption control, environmental control, and follow-up. Higher scores in implemented activities were observed in hospitals that had passed from the registration phase (x- = 26.6; standard deviation [SD] = 16.7) to subsequent phases (x- = 59.7; SD = 14.1), with a statistically significant difference in the overall score (p = 0.05). CONCLUSIONS: The prevalence of smoking in the Catalan Network of Smoke Free Hospitals remains high. However, the prevalence was lower among professionals serving as role models (physicians and nurses) than among the general population. The project's implementation increased according to the time since enrollment in the network, especially in the first year. PMID- 17915121 TI - [Analysis of risk assessment scales for pressure ulcer]. AB - OBJECTIVE: To perform a literature review of the scales and instruments used to assess the risk of developing a pressure ulcer (RPU) in the adult and elderly population and to analyze whether these scales meet the criteria of validity and reliability. To determine whether a specific scale has been developed for use in the domiciliary care setting and adapted to the Spanish environment. METHOD: We performed a descriptive study with a search of the CUIDEN, IME, CINAHL and MEDLINE databases between January 1990 and December 2005. A specifically-designed form was used to register the variables. Data extraction was performed by a single person. The key words used were pressure ulcer, decubitus ulcer, pressure sore, risk evaluation scales, validity, sensitivity, specificity and reliability and their equivalents in Spanish. RESULTS: Complete descriptions of 22 instruments and studies of the validity and/or reliability of 15 instruments were found. CONCLUSIONS: At present, and based on the results of this review, the RPU scale that has shown the greatest validity and reliability is the Braden Bergstrom scale. PMID- 17915122 TI - [Variability in smoking experimentation and risk factors in 4 secondary schools in Murcia (Spain)]. AB - OBJECTIVE: To identify patterns of tobacco experimentation and consumption in 4 geographically close groups of students in the first year of secondary education, as well as attitudes and consumption in their social environments. To identify the factors associated with tobacco experimentation and consumption in each of the student groups. MATERIAL AND METHOD: An observational, descriptive, cross sectional, multicenter study was conducted in 4 secondary schools in the region of Murcia (Spain). The study population consisted of first graders in secondary education, recruited in January 2005. The study variables were collected using a modified version of the FRISC questionnaire. RESULTS: The study population was composed of 377 students (190 boys) with a mean age of 12.6 years (standard deviation [SD] = 0.6). Between 15.3% and 42.1% of the students had smoked at some time. Between 2% and 6.9% smoked regularly. These differences were related to socioeconomic characteristics ("living with the mother", p = 0.049; "living with the father", p = 0.015) and consumption in the environment ("mother", p = 0.013; "friends", p < 0.001; "teachers", p < 0.001). Multivariate analysis revealed an association between experimentation and receiving more than 6R per week pocket money, consumption among friends, experimenting with alcohol and not living in one of the towns studied. CONCLUSIONS: Significant differences were found among the towns studied. Application of standard preventive programs may prove ineffective unless they are adapted to the characteristics of the specific school. PMID- 17915123 TI - [Criteria valued in the nurse selection process in the health area of Lleida (Spain)]. AB - OBJECTIVE: To determine the factors valued by persons taking part in the process of selecting and hiring university nursing graduates. An additional aim was to identify how the need for new specialties is evaluated. METHOD: A cross-sectional study was carried out. From April 2005 to April 2006 we contacted all persons involved in the selection and hiring of nursing graduates in the health area of Lleida (Spain) and gave them a self-administered questionnaire. The questionnaire included items on training, other aspects unrelated to training and issues related to the activation of new specialties. RESULTS: Thirty-four persons involved in the selection and hiring process (89.5%) believed that the most important factor was the training profile. High scores were assigned to attitudes (6/6), competence in drug administration (5.8/6), and consideration of emotional, physical and personal care (5.7/6). Ninety percent of those interviewed agreed on the current need for the new law on specialties. All the specialties obtained a high score based on current need. The 2 most highly scored specialties were those already developed (mental health and midwifery). CONCLUSIONS: The most highly valued aspect of training was attitudes. This finding should be kept in mind in the training of university nursing graduates. Competencies such as participation in teaching and directing or coordinating groups are insufficiently valued. The need to activate all the specialties is positively valued. PMID- 17915124 TI - [Leeches in the intensive care unit: nursing care]. AB - Leeches have been used in medicine since ancient times. Leeches or Hirudos were used to treat multiple diseases, since the bleeding they induce was related to purification. This practice subsequently fell into disuse until the 1980s when leeches again began to be applied in the treatment of venous congestion and in plastic and reconstructive surgery, especially in the USA and Germany. The use of leeches is not yet widespread in Spain. One of the Spanish hospitals pioneering this practice is the University Hospital Joan XXIII de Tarragona, where leeches are employed in the Maxillofacial Service among patients with microvascularized grafts. The present article describes the therapeutic use of leeches (Hirudo medicinalis), as well as the nursing care and complications in patients undergoing this treatment. The aim is to ensure that this new technique is used with maximal safety and quality. PMID- 17915125 TI - [Nursing care in patients undergoing radiological surgery. A case report]. AB - We report the case of a 73-year-old man with medical diagnoses of long-standing diabetes mellitus, chronic ischemia of the lower limbs and intermittent claudication, for which the patient had been treated with minimally invasive radiological surgery. On arrival at the radiology unit, the patient had nursing diagnoses of anxiety and fear. Intraoperatively, the client had nursing diagnoses of pain, urine retention and infection risk. At discharge, a collaboration problem was detected and hemorrhagic risk. The patient received individualized nursing care. Interventions were planned following the nursing intervention classification (NIC) and the expected results for these interventions followed the Nursing Outcomes Classification (NOC) taxonomy. The application of an appropriate nursing care plan contributes to making the patient's hospital stay easier, more comfortable and less traumatic. PMID- 17915126 TI - [Do decisions on hospital nurse staffing influence patient safety?]. PMID- 17915127 TI - [Hospital organizational configuration is the key to success in attracting and retaining nurses]. PMID- 17915128 TI - [On "Nursing care interventions in the prevention of ventilator-associated pneumonia"]. PMID- 17915129 TI - [Efficacy and safety of a reduced-dose of stavudine in HIV-infected patients under immunological and virological stable conditions]. AB - BACKGROUND AND OBJECTIVE: Stavudine (d4T) has shown a favourable short and long term tolerability profile. Nevertheless, its usage is currently decreasing due to some safety concerns. We aimed to evaluate the efficacy and safety of d4T low dose-based regimens. PATIENTS AND METHOD: This was a multicenter and retrospective review chart of patients receiving standard doses of d4T for > or = 6 months (weight > 60 kg: 40 mg/12 h; weight < 60 kg: 30 mg/12 h) and having undetectable viral load for at least 3 months before the d4T dose reduction (weight > 60 kg: 30 mg/12 h; weight < 60 kg: 20 mg/12 h). Immunological and viral parametres, lipid profile and side effects were determined. RESULTS: A total of 982 patients were included. The main reason for reducing the dose was prevention of toxicity (76%). After 6 months of follow-up, 97% and 84% patients had less than 400 and 50 cp/ml, respectively, and the CD4 cell count increased by 38 cel/ml. Lipids, lipodystrophy and peripheral polineuropathy improved but there was no statistical significance. CONCLUSIONS: A d4T dose reduction in an immuno virologically stable population does not affect treatment efficacy. Longer follow ups are required to confirm improvements in the safety profile. PMID- 17915130 TI - [Incidence and mortality of massive transfusion in a university hospital: study of the period 2001-2005]. AB - BACKGROUND AND OBJECTIVE: There are few epidemiological studies on massive transfusion (MT), although they may be important to evaluate possible strategies to reduce the number of transfused units, as well as transfusion side-effects. We, therefore, retrospectively assessed the incidence of MT at our institution (a 700-bed university hospital) during a 5-year period. PATIENTS AND METHOD: Local blood bank records were searched for MT episodes occurred from January 2001 to December 2005. MT was defined as the transfusion of 8 or more packed red cell (PRC) units within 24 h. Patient's clinical data were exclusively gathered from the blood requesting form. RESULTS: Overall, 304 episodes of MT were identified in 288 patients (one episode per week), who received 4,845 PCR units (3,515 units within the first 24 h), because of ruptured aortic aneurism (n = 62), poly-trauma (n = 57), upper digestive bleeding (n = 51), cardiac surgery (n = 41), elective surgery (n = 36), emergency surgery (n = 30), and oncology surgery (n = 27). Mortality rate was 48%, and multivariate analysis identified age (odds ratio [OR] =1.023; 95% confidence interval [CI]. 1.006-1.040) and number of PRC transfused within the first 24 h (OR = 1.094; 95% CI, 1.0032-1.160) as weak but significant independent predictors of mortality, whereas poly-trauma diagnosis was a protective factor (OR = 0.325; 95% CI, 0.112 - 0,940). CONCLUSIONS: Overall, the mortality rate among patients receiving MT was very high, and was influenced by the number of transfused units, patient's age, and admitting diagnose. As the majority of the MT episodes occurred within the surgical or polytrauma context, possible strategies to reduce the volume of MT are discussed. PMID- 17915131 TI - [Utility of amylase levels in malignant pleural effusions]. AB - BACKGROUND AND OBJECTIVE: To analyze the utility of the measurement of pleural amylase levels (AL) and pleural fluid/serum amylase ratio (AR) in malignant pleural effusions. PATIENTS AND METHOD: Prospective and comparative study of AL and its AR in relation to the patient and pleural fluid characteristics in 295 malignant effusions and 673 nonmalignant. RESULTS: There were 103 patients with AL greater than 100 U/l (11%) and 268 with AR greater than 1 (28%): 53 (18%) and 109 (37%) in malignant effusions respectively. Patients with malignant effusions had higher AL and AR, especially when tumour origin was lung cancer, had positive pleural citology or biopsy and showed an adenocarcinoma. Multivariate regression analysis revealed a significant difference in the changes in AL associated with positive pleural citology or biopsy and massive pleural effusion. The malignant effusions had higher AL in lung cancer of stage IV. CONCLUSIONS: AL and AR should not be routinely measured to exclude a malignant effusion. A high AL or AR was related to positive pleural citology or biopsy, a massive pleural effusion and lung cancer with an advanced disease. PMID- 17915132 TI - [Stavudine in antiretroviral therapy]. PMID- 17915133 TI - [New therapies in psoriasis]. PMID- 17915134 TI - [Fannin-Lubbock II hemoglobin [beta111(G13)Val -> Leu y beta119(GH2)Gly -> Asp]: description of 4 new cases]. AB - BACKGROUND AND OBJECTIVE: The aim of this work is the molecular description of a 4 new cases of the hemoglobin Fannin-Lubbock II, which presents the substitution of 2 amino acids in the same beta globin chain. PATIENT AND METHOD: Four cases belonging to 3 families all of white race and from Spain are studied. The molecular analysis was done with the sequence of the products of amplification for polymerase chain reaction of the beta globin gene. RESULTS: The molecular study demonstrated the mutation GTC -> CTC in the codon 111 that determines a change of valine for leucine and the mutation GGC -> GAC in the codon 119 that determines a change of glycine for aspartic acid, both in heterozygote state. CONCLUSIONS: Until the present time 17 hemoglobin variants have been described with 2 substitutions of amino acids in the beta globin chain. The hemoglobin Fannin-Lubbock II had been described in 5 Spanish families; therefore, the communication of these new cases suggests it could be a haemoglobin variant relatively frequent and circumscribed to our population. PMID- 17915135 TI - [Lessons about recombinant activated factor VII. Ten years since its registration for use in hemophilia complicated with inhibitor]. PMID- 17915136 TI - [Psychological treatment for obesity]. AB - Obesity is a chronic disease. Current management is based in the modification of the lifestyle, mainly regarding to eating habits and physical activity. Eating habits are acquired during the childhood and kept through the entire life. Modification of any habit requires the use of specific psychological methods such as cognitive-behavioural therapy, which is based on first notions of learning theories. Very often, obese patients and therapist think that the obesity is a problem not related to other aspects of their lives. Thus the objective of the treatment is only weight loss, instead of looking for a modification of patient's behaviour. In the present review we try to update the treatment of obesity in adult patients mainly regarding to the psychological approach. PMID- 17915137 TI - [Subacute impairment in an elderly patient with dementia]. PMID- 17915138 TI - [Radiodermatitis after fluoroscopy-guided diagnostic and therapeutic procedures]. PMID- 17915139 TI - [Aerosolized colistin in the treatment of multidrug-resistant Acinetobacter baumannii and Pseudomonas aeruginosa pneunomia]. PMID- 17915140 TI - [Severe hyperkalaemia caused by ginger in cirrhotic patient]. PMID- 17915141 TI - [Subacute thrombosis of drug-eluting-stents in a patient with a mechanical aortic prosthesis: the importance of dual antiplatelet therapy]. PMID- 17915142 TI - [Hansen's disease]. PMID- 17915143 TI - [Magnetic resonance imaging in the evaluation of congenital intracardiac shunts]. PMID- 17915144 TI - [Heart failure management programs: in favour of universal implementation]. PMID- 17915145 TI - [C-Reactive protein and restenosis of drug-eluting stents. Intravascular ultrasound solves the mystery]. PMID- 17915146 TI - [Cardiovascular magnetic resonance evaluation of left-to-right shunts due to cardiac septal defects in patients with pulmonary arterial hypertension]. AB - INTRODUCTION AND OBJECTIVES: As cardiac septal defects are frequently associated with pulmonary arterial hypertension, hemodynamic assessment is essential before deciding on surgery. The aim of this study was to evaluate the use of cardiovascular magnetic resonance imaging for assessing cardiac shunts and for quantifying pulmonary artery systolic pressure in patients with cardiac septal defects. METHODS: This cross-sectional study involved patients with cardiac septal defects and clinically suspected severe pulmonary arterial hypertension who had an indication for cardiac catheterization and in whom magnetic resonance imaging was not contraindicated. Each test's results were evaluated independently by two expert radiologists and interventional cardiologists who were blinded to the results of the other test. The procedures were compared using confidence limits and intraclass correlation coefficients. RESULTS: The study involved 29 patients (18 female and 11 male) aged from 30 days to 18 years; seven had an atrial septal defect, 14 had a ventricular septal defect, and eight had an atrioventricular septal defect. The correlation coefficients for measurements made using the two procedures were 0.80, 0.75, 0.81 and 0.58 for pulmonary output, systemic output, flow ratio, and systolic pressure in the pulmonary artery, respectively. Cardiovascular magnetic resonance tended to underestimate systemic output by 0.80 L/min, pulmonary output by 1.35 L/min, left-to-right shunt flow by 0.12 L/min, and systolic pressure in the pulmonary artery by 16.5 mmHg. The complication rate with cardiac catheterization was 31% compared with 3.4% with cardiovascular magnetic resonance imaging. CONCLUSIONS: The evaluation of patients with cardiac septal defects and pulmonary arterial hypertension should initially be performed using noninvasive diagnostic techniques. PMID- 17915147 TI - [Randomized controlled clinical trial of a home care unit intervention to reduce readmission and death rates in patients discharged from hospital following admission for heart failure]. AB - INTRODUCTION AND OBJECTIVES: To determine the effectiveness of a primarily educational intervention in heart failure (HF) patients implemented in a home care unit. METHODS: This randomized controlled clinical trial involved 279 HF patients who were discharged from a tertiary-care hospital between February 2001 and June 2002. Patients with dementia, terminal non-cardiac disease, or chronic obstructive pulmonary disease were excluded. Data collected included the cause of cardiac decompensation. A primarily educational intervention was implemented in the patient's home for up to 15 days after hospital discharge. Treatment was adjusted during the first week if necessary. The primary outcome measure was the 1-year cumulative incidence of readmission or death. Secondary measures were the incidence of readmission, mortality, and emergency department admission. Telephone interviews were carried out 3, 6 and 12 months after discharge, and clinical records were updated when necessary. Emergency department admission in the first 6 months was monitored. RESULTS: At 1-year follow-up, 62 of the 137 patients (45.3%) in the intervention group had been readmitted or died, compared with 75 of the 142 (52.8%) in the control group, (relative risk=0.86, P=.232). Among patients who suffered decompensation because failure to adhere to treatment, 16 of the 45 (35.6%) in the intervention group were readmitted or died, compared with 34 of the 56 (60.7%) control group patients (relative risk=0.59, P=.016). CONCLUSIONS: This intervention is feasible but, when applied indiscriminately to every discharged heart failure patient, the best that can be expected is only a modest reduction in readmission and death rates, which, in this study in particular, did not achieve statistical significance. PMID- 17915148 TI - [Relationship between plasma C-reactive protein level and neointimal hyperplasia volume in patients with zotarolimus-eluting stents. Volumetric analysis by three dimensional intracoronary ultrasound]. AB - INTRODUCTION AND OBJECTIVES: C-reactive protein (CRP) is an inflammatory marker that predicts cardiac events in patients with coronary syndromes. However, data on the relationship between the CRP level and in-stent restenosis are contradictory. The objective of this study was to investigate the relationship between the basal CRP level and the neointimal hyperplasia volume measured by intracoronary ultrasound 4 months after implantation of a zotarolimus-eluting stent. METHODS: The study included 40 consecutive patients who underwent zotarolimus-eluting stent implantation. Patients were divided into quartiles according to their preprocedural CRP level. Intracoronary ultrasound was performed after stent implantation and at 4 months, and the neointimal hyperplasia volume was determined using Simpson's rule. Correlation and linear regression analyses were used to evaluate the relationships between variables. Multivariate analysis was used to identify variables that were independently related to neointimal hyperplasia volume. RESULTS: The patients' mean age was 58 (8) years, 55% were male, and 40% had diabetes mellitus. There was no difference in baseline characteristics between the quartiles. The hyperplasia volumes were 4.8 (4.2) microl and 15.8 (10.0) microl in the first and fourth quartiles, respectively (P< .001). There was a significant positive correlation between the CRP level and neointimal hyperplasia volume (r = 0.64, P=.0001). The CRP level, the postimplantation lumen volume, and the final deployment pressure were all independent predictors of neointimal hyperplasia. CONCLUSIONS: In this study, an independent correlation was observed between the CRP level before zotarolimus eluting stent implantation and the neointimal hyperplasia volume at 4-month follow-up. PMID- 17915149 TI - [Changes in percutaneous coronary intervention over the last 20 years: demographic, epidemiologic and clinical characteristics, techniques, and outcomes]. AB - INTRODUCTION AND OBJECTIVES: The aim of this study was to describe the changes observed in clinical practice in our interventional cardiology unit over the last 20 years. METHODS: Between January 1, 1986 and December 31, 2005, >or=17,204 percutaneous transluminal coronary angioplasties (PTCAs) were performed at our center. They were analyzed in four periods of 5 years each. After each procedure, prospective data on patient, procedural, and outcome variables were recorded. The data were analyzed with regard to when the procedure was performed, and the patients' sex and age (i.e., < or >or=75 years). Data from 2006 were used as a reference, but were not included in the analysis. RESULTS: The number of PTCAs increased significantly. Over the time period, mean patient age increased (from 57[10] years to 62[12] years for males and from 66[10] years to 70[11] years for females), and there were significant increases in the proportions of women (from 16% to 22%) and patients aged >75 years (from 7% to 22%). Among men, the incidence of smoking decreased while that of hypertension, diabetes and hyperlipemia increased. Among women, however, there was no change in risk factors. The numbers of urgent and emergent procedures (17% of PTCAs were for acute myocardial infarction in 2006) increased in both sexes and age groups, more ad hoc procedures were carried out, and more lesions were treated, usually with a stent. Over time, the success rate increased and the complication rate decreased in both sexes and age groups. In-hospital mortality for all procedures, except those for acute myocardial infarction, was 0.7% in males and 1.4% in females (P=.012). CONCLUSIONS: Significant changes were observed over the last 20 years in the baseline characteristics of, the techniques used in, and the outcomes obtained in patients undergoing percutaneous coronary intervention. PMID- 17915150 TI - [Predictors of improved left ventricular systolic function after surgical revascularization in patients with ischemic cardiomyopathy]. AB - INTRODUCTION AND OBJECTIVES: Although it is known that the presence of myocardial viability predicts an increase in ejection fraction after revascularization in patients with ischemic cardiomyopathy, little is known about other predictive factors. The aim of this study was to identify variables that can predict an increase in ejection fraction after coronary revascularization surgery in patients with ischemic cardiomyopathy and a viable myocardium. METHODS: The study included 30 patients (mean age 61.6 [11] years, one female) with ischemic cardiomyopathy (ejection fraction or=5% occurred after surgery in 17 of the 30 patients (56.6%). These patients were characterized by the presence of left main coronary artery disease (P< .004), a large number of grafts (P< .03), a high perfusion summed difference score (P< .012), a low end-diastolic volume (P< .013), and a low end-systolic volume (P< .01). An end-systolic volume <148 mL and a summed difference score >or=4 gave the best predictive model (P=.001, R2=0.73) for an increase in ejection fraction. CONCLUSIONS: In patients with ischemic cardiomyopathy and a viable myocardium, the main determinants of an increase in ejection fraction after revascularization surgery were low levels of left ventricular remodeling and myocardial ischemia. PMID- 17915151 TI - [Cardiovascular responses to hypertonic NaCI injection into the anteroventral third ventricle region in rats with fructose-induced hypertension and insulin resistance]. AB - INTRODUCTION AND OBJECTIVES: To investigate the hemodynamic sympathetic response evoked by NaCI microinjection into the third ventricle anteroventral brain area (AV3V) in rats long-term fed with high fructose diet. METHODS: Twelve male rats received 60% fructose enriched diet for 6 months. Control rats (n=12) received regular diet. RESULTS: Fructose diet increased (P< .01) body weight; plasma glucose, triglycerides; cholesterol, insulin; systolic (SBP) and diastolic blood pressure (DBP). Basal heart rate (HR) did not change. AV3V microinjection of 2 microL of hypertonic 1.5 M NaCI in fructose fed rats increased SBP 44.64(3.6) mm Hg, DBP 19.9(2.4) mm Hg and HR 66.2(8.4) beats/min over basal values (P< .01). In control rats, smaller responses were observed, SBP increased 28.33(3.10) mm Hg, DBP 13.0(1.9) mm Hg and HR 23.0(5.0) beats/min over basal values (P< .01). CONCLUSIONS: Long-term fructose diet in rats induces cardiovascular hyperactivity of AV3V neurons to sodium chloride, and is associated to hypertension and insulin resistance. PMID- 17915152 TI - [Metalloproteases, vascular remodeling and atherothrombotic syndromes]. AB - Defects in the synthesis and breakdown of the extracellular matrix (ECM) are now seen as key processes in the development of atherosclerosis and its thrombotic complications. Correlations have been observed between circulating levels of ECM biomarkers and the clinical manifestations of and risk factors for atherosclerosis. Several matrix metalloproteinases (MMPs), endopeptidases that can degrade the ECM, such as MMP-9 and MMP-10, play important roles in the pathophysiology of atherothrombosis and contribute to the expansion of abdominal aortic aneurysms. Moreover, they may also be useful biomarkers of atherosclerotic risk and serve as predictors of coronary and cerebrovascular disease recurrence. Although at present the effect of tissue inhibitors of MMPs (TIMPs) on cardiovascular disease prognosis is still uncertain, the ECM could be a promising therapeutic target in atherothrombotic disease, and several MMP inhibitors are currently undergoing clinical trials. PMID- 17915154 TI - [Peripheral artery disease: pathophysiology, diagnosis and treatment]. AB - Peripheral artery disease is one of the most prevalent conditions, and it frequently coexists with vascular disease in other parts of the body. Early diagnosis is important for improving the patient's quality of life and for reducing the risk of serious secondary vascular events such as acute myocardial infraction (AMI) or stroke. The best noninvasive measure for identifying the presence of occlusive arterial disease is the ankle-brachial index, which can also be used to indicate the prognosis of the affected extremity and to predict the likelihood of AMI during follow-up. Intermittent claudication in the lower limbs is the most common clinical presentation. The presence of critical ischemia (i.e., with rest pain or trophic changes) indicates the need for prompt revascularization because of the high risk of limb amputation. The more proximal the affected arterial segment, the better the outcome of the procedure. Endovascular treatment is usually reserved for lesions affecting multiple segments. It gives poorer results in occluded arteries. In extensive disease, conventional surgery is usually the best option. PMID- 17915155 TI - [Biventricular support and cardiectomy]. PMID- 17915153 TI - [ESH/ESC 2007 Guidelines for the management of arterial hypertension]. PMID- 17915157 TI - [Safety of diagnostic transradial catheterization in patients undergoing long term anticoagulation with coumarin derivatives]. AB - The aim of this study was to evaluate the safety of transradial catheterization without prior withdrawal of oral anticoagulation. In total, 183 patients on long term acenocoumarol treatment who underwent transradial catheterization were included in a prospective study. In 135 patients, the coumarin derivative was continued without interruption before and during catheterization and, in 48, the drug was stopped 48 hours in advance. No severe thromboembolic or hemorrhagic complications occurred. At 7-day follow-up, the rate of occurrence of hematomas >3 cm in size was 5.5% in the group that continued anticoagulant treatment, and 10.4% in the group that stopped oral anticoagulation (P=.31). The transradial approach appears to be a safe option for patients receiving long-term coumarin treatment and could be the technique of choice because it avoids the drawbacks and complications associated with the withdrawal of oral anticoagulation. PMID- 17915156 TI - [Initial experience with the Impella left ventricular assist device for postcardiotomy cardiogenic shock and unprotected left coronary artery angioplasty in patients with a low left ventricular ejection fraction]. AB - Low-output syndrome is one of the leading causes of death following open-heart surgery or high-risk angioplasty. Ventricular assist devices have been used to treat patients who suffer from postoperative cardiogenic shock despite use of an intraaortic balloon pump and maximum inotropic support. The Impella pump (Impella Cardiosystems AG, Aachen, Germany) is a newly introduced left ventricular assist device that has been shown to reduce infarct size and to accelerate recovery of stunned myocardium. We report our initial experience using the Impella device for the treatment of cardiogenic shock following cardiopulmonary bypass and for maintaining hemodynamic stability in high-surgical-risk patients undergoing unprotected left coronary artery angioplasty. PMID- 17915158 TI - [Limitations in the Clinical Assessment of Obesity: comments on the American Heart Association's 2006 Statement]. PMID- 17915160 TI - [Cardiac shotgun pellet in the interventricular septum]. PMID- 17915159 TI - [Ventricular tachycardia induced by exercise testing in a patient with Brugada syndrome]. PMID- 17915161 TI - [Coronary aneurysms of the left anterior descending coronary artery and the right coronary artery after drug-eluting stent implantation]. PMID- 17915162 TI - A randomized trial comparing the efficacy and tolerability of two HAART strategies at two years in antiretroviral naive patients. AB - BACKGROUND: The use of HAART combining 2 nucleoside analogues reverse transcriptase inhibitors (NRTIs) plus one protease inhibitor (PI) or 2 NRTIs + 1 non-nucleoside reverse transcriptase inhibitor (NNRTI) has shown comparable efficacy. The study was designed to compare long term (2 years) effectiveness of two antiretroviral (ARV) treatment strategies in patients not previously treated: starting with a nelfinavir based HAART switching to nevirapine in case of failure or side effects or the reverse sequence. METHODS: This multicenter, randomized, open label clinical trial enrolled ARV-naive HIV patients with CD4 counts below 500 cells/mm3. They were randomly assigned to start ddI + d4T + nelfinavir (switching to ZDV + 3TC + NEV in case of failure or toxicity) (PI-NEV arm) or ddI + d4T + nevirapine, switching to ZDV + 3TC + NFV in case of failure or toxicity (NEV-PI arm). The primary study endpoint was the Kaplan-Meier estimates of the time to failure after switching to second regimen if necessary (considering failure as two consecutive plasma HIV-1 RNA determinations above 200 copies/mL, death, a new category C event or toxicity leading to treatment discontinuation of the second regimen) after a minimum follow-up of two years. RESULTS: A total of 137 patients were evaluable (67 and 70 in the PI-NEV and NEV-PI arms respectively). Baseline characteristics did not differ among groups. Kaplan-Meier estimates of time to failure did not show differences between the two arms neither in the on-treatment (OT) analysis (log rank test, p = 0.81) nor in the intent-to-treat (ITT) analysis (p = 0.58). At 24 months, the estimated proportion of patients free of failure were 72% and 66% respectively in the PI-NEV and NEV PI arms OT analysis (p = 0.54) and 73% and 64% in the PI-NEV and NEV-PI arms in the ITT analysis (p = 0.49). The difference in the median in CD4+ lymphocyte count at 24 months was not significantly different in the two groups: 393 and 307 CD4 cells/mm3 in the PI-NEV and NEV-PI arms respectively (p = 0.167). The incidence of adverse events (AEs) in the two arms was very similar: 50 (75%) in the PI-NEV and 54 (70%) in the NEV-PI group, as it was for grade 3-4 AEs leading to drug switching. CONCLUSION: At two years both treatments strategies (PI-NEV vs NEV-PI) had a high and comparable efficacy and were generally well tolerated. PMID- 17915163 TI - [Differences in sociodemographic, clinical, psychosocial and health care characteristics between men and women diagnosed with fibromyalgia]. AB - INTRODUCTION: Health differences between men and women are determined by biological differences although health services often contribute to gender inequalities. Very few studies that analyze gender differences have been made up to date in these patients. This study aims to analyze sociodemographic, clinical and psychosocial differences between men and women diagnosed with FM and to examine the differential impact of their symptoms on their usual activities, including work environment, and the response these patients obtain from the health care system. MATERIAL AND METHODS: A descriptive cross-sectional survey was carried out with all the patients diagnosed with FM in 2003 in three clinics rheumatology units of a university hospital in Spain. RESULTS: The sociodemographic characteristics were very similar in men and women. However, there was a greater proportion of men diagnosed with FM on sick leave, compared to women with the same diagnosis. Men had a worse perception of their health, a higher percentage of psychiatric history and current mental illness and more impact of the disease. DISCUSSION: This is one of the first studies in Spain examining the differences between men and women diagnosed with FM. The results obtained in this study corroborate that, as in other diseases, there are gender differences in the clinical and psychosocial characteristics of men and women diagnosed with FM. PMID- 17915164 TI - [Renal failure and insulin resistance: effect of the dialysis dose]. AB - BACKGROUND: The main cause of death in patients undergoing hemodialysis are cardiovascular events. We have analyzed the association between intradialysis fractional clearance of urea or Kt/V index in patients with chronic renal failure in hemodialysis as an indicator of adequate dialysis and the classical factors of cardiovascular risk. PATIENTS AND METHODS: A total of 47 patients with chronic renal failure on hemodialysis were included. Diabetes mellitus was considered an exclusion criteria. Optimization of dialysis was analyzed by Kt/v index in accordance with the Daugirdas formula. The cardiovascular risk factors evaluated were: insulin resistance, calculated according to the HOMA method, total cholesterol, LDL-cholesterol, triglycerides, arterial hypertension, obesity and metabolic syndrome. The relationship between cardiovascular risk factors and Kt/V index was analyzed with the variant and multivariant analysis. RESULTS: The HOMA median (interquartile range) of the patients was 1.16 (0.53-5.77). HOMA was correlated with triglycerides and HDL-cholesterol levels. HOMA was significantly greater in those who had less adapted dialysis (Kt/V < 1.4) (1.9 +/- 1.3 vs 1.0 +/- 0.3, p = 0.02). Furthermore, a negative correlation was found between HOMA and Kt/V. The multivariant analysis of the factors associated to HOMA demonstrated that the only associated parameters were Kt/V greater than 1.4, body mass index and age. CONCLUSIONS: In patients with chronic renal failure, adapted dialysis, determined by the Kt/V index, correlated negatively with insulin resistance. Based on these data, we suggest the need for longitudinal studies that consider this index as a predictive variable of cardiovascular events in this type of patients. PMID- 17915165 TI - [Reproducibility of the glucose tolerance test in patients with impaired fasting glucose]. AB - BACKGROUND AND OBJECTIVES: Impaired fasting glucose (IFG) is defined by a fasting glucose between 5.6 and 6.9 mmol/l in subjects with no known diabetes. The present study objectives were: a) to analyze the glucose tolerance test (GTT) reproducibility and b) to assess this test's diagnostic classification agreement. PATIENTS AND METHOD: Cross-sectional study in adult patients diagnosed with IFG. Study subjects underwent a 75 g GTT in two occasions. RESULTS: Fifty-nine patients were studied. The interval between GTT tests was 37 +/- 26 days. Fasting and post-GTT plasma glucose intra-individual variation coefficients were 6.9 and 31.0%, respectively. Diagnostic agreement between the two tests (normal tolerance vs. abnormal tolerance) was measured using the kappa index: 0.62 (95% CI 0.42 0.82). Agreement ranged from 80% (95% CI, 70-90%) to 83% (95% CI, 73-93%) depending on whether the first GGT results were abnormal or normal, respectively. CONCLUSIONS: GTT reproducibility is moderate in patients diagnosed with IFG. Considering this fact, perhaps this test should be repeated before therapeutic decisions are made. PMID- 17915166 TI - [Biological therapy with anti-TNF antibodies. Extending the spectrum of its indications?]. AB - Tumor necrosis factor (TNFalpha) is a main mediator in the inflammatory answer of many diseases. The anti-TNFalpha antibodies (infliximab, etanercept, adalimumab) block their action, preventing the inflammatory answer and the damage it produces. In this paper, we review the treatment of some diseases in which these drugs are experimentally used. These are: ulcerative colitis, sarcoidosis, graft versus-host disease, adult Still's disease, some systemic vasculitis, systemic lupus erythematosus and Behcet disease. PMID- 17915167 TI - [Which patients with heart failure should be offered cardiac resynchronization?]. AB - Chronic heart failure is a very prevalent disease in developed countries. In recent decades, very important advances in drug therapy have occurred. However, mortality is still very high. One third of patients with a low ejection fraction and New York Heart Association (NYHA) functional class IIII-IV have a wide QRS. This means that there is often resynchronization of contraction and higher mortality. In order to improve the prognosis, the therapy based on cardiac resynchronization device has bee4n shown to be a complementary medical treatment and has contributed to clinical, hemodynamic and mortality improvements. In this article, we aim to show the results of clinical trials and recommendations of the main guidelines regarding this therapy. PMID- 17915168 TI - [Patient safety and quality of care]. AB - Adverse effects related to health care are common and partly avoidable. We need to identify how and why adverse events occur and how system defects may contribute to their occurrence. Systems and processes can be designed to help prevent errors and decrease harm that occurs when they are not intercepted. Tactics to reduce errors and mitigate their adverse effects include reducing complexity and optimizing information processing. Implementation of information technology may offer great promise but the most important is to make an effort to promote a culture of safety. PMID- 17915169 TI - [Clinical-pathological lectures of the SEMI]. PMID- 17915170 TI - [Prevalence of vertebral fractures in hip fracture patients]. AB - BACKGROUND: Fractures are the clinical complication of osteoporosis. There are no previous studies that describe the prevalence of vertebral fractures (VF) in patients admitted into a hospital due to a hip fracture (HF). OBJECTIVE: To study the prevalence of vertebral fractures in elderly women in the moment of their admission to the hospital due to a hip fracture. METHOD: This is a cooperative, multicentric, case-control study, performed in 21 different hospitals of Spain by the Working Group on Osteoporosis of the Spanish Society of Internal Medicine. A total of 143 elderly women with hip fractures comprised the case group. The control group consists of 138 elderly women admitted into other wards of the hospital due to other diseases with no relationship with osteoporosis. A questionnaire was administered and a lateral thoracic and lumbar X-ray was performed to assess vertebral fractures applying Genant's criteria. RESULTS: The mean age of the patients with HF was 79.8 +/- 6.9 years and the mean age of the controls was 77.7 +/- 8.9 years. Patients suffering from HF had less weight than controls (BMI: 25.9 +/- 4.4 g/m2 vs 27.7 +/- 5.2 kg/m2, p = 0.002). Prevalence of VF was 62.6% in patients with HF, and 50% in controls (p = 0.039). CONCLUSIONS: Elderly women admitted to a hospital due to hip fracture have a very high prevalence of previously undiagnosed VF. Indeed, elderly women admitted into the hospital because of other diseases also have a high prevalence of VF. These facts must be taken into account due to the morbidity and mortality of VF, that increases the HF morbidity and mortality. PMID- 17915171 TI - [Dypsnea in a patient with liver transplant]. PMID- 17915172 TI - [Spastic paraparesis due to stenosis of dorsal canal caused by Paget's disease]. PMID- 17915173 TI - [Cutaneous nodules in patient with acute myeloid leukemia]. PMID- 17915174 TI - [An unusual cause of multiple pulmonary nodules]. PMID- 17915175 TI - [Satisfaction Survey to the User made in a new unit of the Hospital de Talavera de la Reina]. PMID- 17915177 TI - [Abdominal pain and functional impotence of the low limb as presentation form of an abdominal aorta aneurysm with chronic rupture]. PMID- 17915176 TI - [Endocarditis and aortic abscess due to Corynebacterium macginleyi]. PMID- 17915178 TI - [Intermediate care unit of internal medicine]. PMID- 17915179 TI - [Scurvy: an unusual cause of purpura, ecchymosis and arthralgias]. PMID- 17915180 TI - [Pharmacotherapy of attention-deficit/hyperactivity disorder (ADHD) and comorbid disorders]. AB - Attention-Deficit/Hyperactivity Disorder (ADHD) is one of the most commonly diagnosed disorders in child and adolescent psychiatry. Patients with ADHD show increased risk for comorbid psychiatric disorders such as mood disorders, anxiety disorders, tic disorders and disruptive behavior disorders. Using criteria and methods of evidence-based medicine, in this paper possible algorithms for ADHD and comorbid disorders are presented. In conclusion, these algorithms aim to support health care professionals and can serve as guideline in decision making in pharmacotherapy. Stimulants are first-line medication treatment of ADHD without comorbid disorder. In co-existing aggression, major depressive disorder and tic disorder treatment with stimulants should be initialized at first. Only in case of unchanged persistence of comorbid symptoms specific pharmacological therapy should be combined. In comorbid anxiety disorders atomoxetine can be used as first-line treatment. PMID- 17915181 TI - [The effects of a computer assisted cognitive training on neuropsychological parameters, mood and dysfunctional cognitions in depressive patients]. AB - AIM: In this pilot study a computer-assisted cognitive training programme (CAT) was tested and evaluated in patients with depressive syndromes for its effects on neuropsychological parameters, modd and dysfunctional cognitions. METHODS: A total of 48 patients were randomized either to the treatment or the control group. The former underwent 20 sessions, 30-45 minutes each, of computer-assisted cognitive training, twice a week. At the beginning and at the end of the training (ten weeks later), both groups were given neuropsychological tests (memory, attention, concentration and information processing speed) and also questionnaires to assess depressive mood, thoughts, negative self communication and locus of control. RESULTS: At the start of the study, both the groups information processing speed. All these were absent in the control group. Also the depressive mood, as well as negative self-communication markedly improved in the training group. As expected, these results were not observed in the control group. CONCLUSION: According to these results in depressed patients a computer assisted cognitive training programme might have a positive influence not only on their neuropsychological functions but also on their dysfunctional beliefs. Thus CAT should be part of the treatment of depressive patients. PMID- 17915182 TI - [Unjustly neglected: siblings of people with a schizophrenic psychosis]. AB - OBJECTIVE: Siblings of individuals suffering from schizophrenia are an underrepresented group in research focussing on the needs of carers and relatives of psychiatric patients. The present study aims to investigate differences between siblings and parents as well as spouses, as regards help seeking, utilisation of an open group for relatives, their subjective burden and quality of life. METHODS: 147 relatives of in-patients and patients attending a day hospital where assessed using the General Health Questionnaire (GHQ), the Family Problem Questionnaire (FPQ), the WHO Quality of Life-BREF (WHOQOLBREF) and a questionnaire inquiring about the relatives' utilisation of various sources of information and help throughout the course of the illness. RESULTS: Siblings reported less contact to the patients compared to the two other groups. However, their subjective burden was comparable to that of spouses, who were the group with the highest amount of contact. Siblings' quality of life showed by far less impairment than that of spouses and parents. They reported significantly less utilisation of any source of information and help and were far less likely to be invited to the group for relatives. CONCLUSIONS: Siblings of patients with schizophrenia are a particularly neglected group regarding support aimed at relatives. They are heavily distressed, yet there is little offer of professional support for them. It seems indicated to draw increased attention to this specific group of relatives. PMID- 17915183 TI - [Scuba diving -- a therapeutic option for patients with paraplegia]. AB - OBJECTIVE: Spasticity is often a handicap in paraplegics and interferes with quality of life. Medical therapeutic options (e.g. baclofen, tizanidin) lead to drowsiness, fatigue and loss in activity. On the other hand paraplegics are increasingly active in daily life and leisure (paralympics). Neurorehabilitation is effective in reduction of spasticity, gaining motor function and enhancing quality of life. Hippotherapy (Lechner et al 2003) and aquatic rehabilitation are additive methods. Already 15 years ago Madorsky et al pointed out SCUBA diving as a positive neurorehabilitation procedure. The study group around Stanghelle reported also beneficial aspects on spasticity of patients with spinal cord injuries. These references inspired to introduce a prospective study. METHODS: After obtaining an ethic votum and evaluation assessment for diving permission 6 volunteers with paraplegia entered the pilot study. Medication was kept stable throughout the study time. Supervised by diving instructors and a diving trained doctor the volunteers dived to a platform in the depth of 7.2 meters. The daily diving time was exactly 30 minutes. Stabilized on the platform physiotherapeutic assessment took place in different positions to reduce spasticity. Ashworth Scale and spasm frequency scale were noted daily and at beginning and end of the study the WHO Quality of life Test had to be completed. For objective reasons a locomat training happened before, within a week after and 4 weeks after the study week. RESULTS: All patients did the daily dives without any difficulties. The statistics included the assessment of day 1 versus day 7 of 5 patients and showed a significant reduction of Modified Ashwoth Scale (p=0.04). Quality of life showed an improvement. CONCLUSION: The improvement rationale can only be supposed. A correlation to the ambient pressure suggests itself. Therefore deeper depths should increase the good spasticity results or manage to achieve those faster. Many questions remain, so further studies are necessary to ascertain the ideal standard options. PMID- 17915184 TI - [Alzheimer drugs for mild cognitive impairment]. AB - Improved diagnostic procedures allow to detect more patients with early stages of Alzheimer's disease (AD) within the increasing old population especially in the setting of specialised Memory Clinics. As no disease modifying or secondary preventive pharmacological treatments are presently available for AD this article discusses whether the established pharmacological treatment strategies for Alzheimer's dementia (DAT) could be beneficial as well for patients with amnestic mild cognitive impairment (MCI) that can be regarded a transitional stage between normal aging and DAT in many patients. The author does not recommend to treat MCI patients with Cholinesterase Inhibitors (ChE-I) or memantine first of all due to the general problems of a low specificity of the MCI concept and the missing data concerning the treatment of MCI patients with memantine. Moreover, ChE-I therapy in MCI showed weak beneficial effects on cognition and the delay of DAT onset whereas it is costly and has a high risk of side effects. Additionally, the pooled galantamin-data suggest an increased mortality compared to placebo. Finally, cortical cholinergic deficits and the corresponding cognitive deficits are commonly not present in MCI. PMID- 17915185 TI - [Peace ... In memoriam Oswald Tschirtner (1920-2007)]. PMID- 17915186 TI - Evaluation of follow-up of therapy with fenbendazole incorporated into stabilized liposomes and immunomodulator glucan in mice infected with Toxocara canis larvae. AB - Anthelmintic activity of benzimidazole carbamate anthelmintics is low against dormant Toxocara canis larvae during late infections in paratenic hosts. The present study was conducted to examine the efficacy of pure fenbendazole, or drug incorporated into sterically stabilized liposomes (SL-FBZ) administered to T. canis-infected mice alone and after its co-administration with the immunomodulator (1-->3)-beta-D-glucan against larvae localized in muscles and brains. Therapy with either drug forms (in total 250 mg/kg in 10 doses) commenced on day 28 post-infection (p.i.) and the efficacy of treatment, examined on day 30 after the last dose of drug, was the highest in groups of mice treated with SL FBZ in combination with glucan (89.5+/-5.8% in the muscles, 66.1+/-8.1% in brains). During 56 days of follow-up after termination of therapy, serum levels of anti-TES IgG antibodies, circulating IgG-TES immune complexes (CIC) as well as IgG antibodies to the most immunogenic part of recombinant myosin antigen of T. canis larvae were investigated. In contrast to anti-TES IgG antibodies, levels of CIC and anti-myosin antibodies were in the linear correlation with the efficacy of treatments beginning from day 38 post-therapy. We also showed that the serum levels of CIC as well as anti-myosin IgG antibodies seem to be the suitable serological markers for the monitoring of progress in larval destruction and TES resorption from the tissues. PMID- 17915187 TI - Topical glaucoma therapy as a risk factor for nasolacrimal duct obstruction. AB - PURPOSE: To investigate a possible association between primary open-angle glaucoma (POAG) and primary acquired nasolacrimal duct obstruction (PANDO). DESIGN: Retrospective, comparative study. METHODS: The study group consisted of 209 consecutive eyes (178 patients) whose lacrimal system had PANDO in patients more than 50 years of age during the 10-year study period. The control group consisted of 183 consecutive eyes (183 patients) that underwent cataract surgery during the same period. The main outcome measures were prevalence of POAG in study and control groups and the effect of topical glaucoma therapy use profile on PANDO prevalence. Medical records of all patients included in the study were reviewed. Data collected included demographic details and history and characteristics of POAG treatment. RESULTS: The prevalence of POAG in the PANDO group (23%) was significantly higher than that of the control group (6%; P < .0001). The average history of POAG was longer in the PANDO group (14.10 +/- 5.59 years) compared with the control group (9.55 +/- 7.23 years; P = .025). The average number of topical glaucoma therapy drugs per glaucomatous eye in the PANDO group (1.58 +/- 0.92) was significantly higher than that of the control group (0.73 +/- 0.90; P = .002). Bilateral nasolacrimal duct obstruction (NLDO) was more common among glaucoma patients in the PANDO group (38.23%) compared with nonglaucoma patients in the same group (11.80%; P = .0002). A significantly higher percentage of glaucoma patients in the PANDO group (69%) were treated with timolol, compared with glaucoma patients in the control group (18%; P = .005). CONCLUSIONS: Chronic use of timolol-containing topical glaucoma therapy preparations in glaucoma patients is associated with an increased risk for the development of NLDO. Large-scale prospective studies are needed to ascertain this association. PMID- 17915188 TI - Lysophosphatidic acid-stimulated interleukin-6 and -8 synthesis through LPA1 receptors on human osteoblasts. AB - Using human osteoblastic SaM-1 cells, we investigated the effects of lysophosphatidic acid (LPA) on the production of interleukin (IL)-6 and IL-8, molecules which are capable of stimulating the development of osteoclasts from their haematopoietic precursors, and examined the signal transduction systems involved in their effect on these cells. These human osteoblasts constitutively expressed endothelial differentiation genes (Edg)-2 and Edg-4, which are LPA receptors. LPA increased gene and protein expression of IL-6 and IL-8 in SaM-1 cells. The expression of IL-6 and IL-8 mRNAs was maximal at 1-3h, and the increase in IL-6 and IL-8 synthesis in response to lysophosphatidic acid (1-10 microM) occurred in a concentration-dependent manner. These increases were blocked by Ki16425, an Edg-2/7 antagonist. In addition, LPA caused an increase in the intracellular Ca(2+) concentration ([Ca(2+)](i)), which was inhibited by pretreatment with Ki16425 or 2-aminoethoxy-diphenylborate (2-APB), an inositol 1,4,5-triphosphate (IP(3)) receptor (IP(3)R) blocker. The pretreatment of SaM-1 cells with U-73122, a phospholipase C (PLC) inhibitor, and 2-APB also inhibited the increase in IL-6 and IL-8 synthesis in response to LPA. These findings suggest that extracellular LPA-induced IL-6 and IL-8 synthesis occurred through Edg-2 (LPA(1) receptor) and the activation of PLC and IP(3)-mediated intracellular calcium release in SaM-1 cells. PMID- 17915189 TI - Ultrastructure of dentine carious lesions. AB - OBJECTIVE: The aim of the present study is to investigate the ultrastructural changes within the different zones of carious dentine and compare those changes with sound dentine. METHODS: Transmission electron microscopy and electron diffraction techniques were used to investigate the effect of caries on the inorganic phase of dentine. Areas of interest were identified with optical and scanning electron microscopes. RESULTS: The results demonstrated that the intertubular mineral crystallites decrease in size as caries lesion progresses. In the transparent zone of carious lesion, both intratubular and intertubular dentine consisted of nano-size apatetic crystallites with smaller size in the former. The intratubular mineral phase in transparent zone was found to be chemically similar to the intertubular dentin. CONCLUSIONS: The study suggests that a 'dissolution and precipitation' mechanism is important in understanding the process of formation of intratubular dentine within the transparent zone induced by caries attack. The observed partial demineralisation of intertubular dentine in transparent zone is discussed in terms of dissolution of tubule microbranches and exposure of intertubular dentin to acids. PMID- 17915191 TI - Probing the structure of the Ff bacteriophage major coat protein transmembrane helix dimer by solution NMR. AB - The transmembrane (TM) segment of the major coat protein from Ff bacteriophage has been extensively studied as an example of dimerization in detergent and lipid bilayer systems. However, almost all the information regarding this interaction has been gained through mutagenesis studies, with little direct structural information being available. To this end solution NMR has the potential to provide new insights into structure of the dimer. In order to evaluate the utility of this approach we have studied a selectively 15N-labeled peptide containing the TM segment of MCP (MCPTM) by solution NMR. This peptide was found to give rise to detergent concentration-dependent spectra that were assigned to monomeric and dimeric forms. The standard free energy of this interaction in SDS was estimated from these spectra and found to be consistent with weak but specific dimerization. In addition, similar spectra could be obtained in beta octyl glucoside with intermolecular paramagnetic relaxation experiments demonstrating a parallel arrangement of TM helices in the dimer. In both detergents backbone chemical shift differences between monomeric and dimeric forms of MCPTM showed that the largest changes occur around its GXXXG motif. The resulting structural model is consistent with observations made for MCP mutants previously characterized in biological membranes, opening the door to detailed structural characterization of this form of MCP. These results also have general implications for the study of weakly interacting TM segments by solution NMR since the use of similar sample conditions should allow structural data to be accessed for oligomeric states from a wide range systems that undergo biologically relevant but weak associations in the membrane. PMID- 17915192 TI - I'll believe it when I can see it: imagery rescripting of intrusive sensory memories in depression. AB - Intrusive sensory memories are a common feature of depression but these are not targeted in standard cognitive treatments. Imagery rescripting of distressing memories has so far been mainly used to treat trauma-related disorders and as a component of the treatment of personality disorders. We propose that this approach might also be effective in treating depression. This paper describes the initial application of imagery rescripting as a stand-alone treatment for two patients with a sole diagnosis of major depressive disorder. The two cases are described in detail and follow-up data are reported. Implications for the cognitive treatment of depression and for our theoretical understanding about the mechanisms of change in cognitive therapy are considered. PMID- 17915190 TI - Stimulus-induced reorganization of tight junction structure: the role of membrane traffic. AB - The tight junction forms a barrier that limits paracellular movement of water, ions, and macromolecules. The permeability properties of this barrier are regulated in response to both physiological and pathophysiological stimuli, and this regulation has been modeled by pharmacological agents. Although it is now known that vesicular traffic plays important roles in tight junction assembly, the molecular mechanisms by which vesicular traffic contributes to tight junction regulation remain to be defined. This review summarizes recent progress in understanding mechanisms and pathways of tight junction protein internalization and the relevance of these to tight junction regulation. PMID- 17915193 TI - Impaired expression and function of breast cancer resistance protein (Bcrp) in brain cortex of streptozocin-induced diabetic rats. AB - The aim of this study was to investigate whether diabetes mellitus (DM) affected breast cancer resistance protein (Bcrp) function and expression in rat brain. 5 week and 8-week diabetic rats were induced by streptozocin (STZ). Bcrp expression and function in brain cortex were assessed by western blot and measuring the brain-to-plasma concentration ratios of two typical substrates prazosin and cimetidine, respectively. The diabetic rats were treated with three different agents insulin, aminoguanidine (AG) and metformin (MET). It was found that the brain-to-plasma ratios of prazosin and cimetidine in diabetic rats were significantly higher than those of control rats, which were dependent on duration of diabetes. Lower levels of Bcrp were found in brain cortex of diabetic rats, which were in parallel with increase of brain-to-plasma ratios. Insulin treatment may attenuate the impairment of Bcrp expression and function induced by diabetes. Aminoguanidine and metformin treatment did not prevent the impairment of Bcrp function and expression in brain cortex of diabetic rats. All results gave a conclusion that STZ-induced DM may induce the impairment of function and expression of Bcrp in brain cortex, and lower levels of insulin may mainly contribute to Bcrp dysfunction in brain. PMID- 17915194 TI - KRIBB3, a novel microtubule inhibitor, induces mitotic arrest and apoptosis in human cancer cells. AB - KRIBB3 (5-(5-ethyl-2-hydroxy-4-methoxyphenyl)-4-(4-methoxyphenyl) isoxazole) inhibited cancer cell growth in vitro and in vivo. Flow cytometry studies showed that KRIBB3 caused cell cycle arrest at the G(2)/M phase and subsequent apoptosis. This was confirmed as accumulation of Cyclin B1 and cleavage of poly(ADP-ribose) polymerase (PARP) were detected. While transient inhibition by KRIBB3 led to reversible mitotic arrest, prolonged exposure to KRIBB3-induced apoptosis. Co-immunoprecipitation experiments showed that KRIBB3 initially induced association of inhibitory Mad2 with p55CDC (mammalian homologue of CDC20), an activator of APC/C (anaphase-promoting complex/cyclosome), suggesting that the mitotic spindle checkpoint was activated by KRIBB3. However, the level of this inhibitory complex of Mad2 with p55CDC was gradually decreased 24 h after KRIBB3 treatment, and was hardly detectable after 48 h, indicating some slipping of the mitotic checkpoint. Consistent with these observations, KRIBB3 activated the mitotic spindle checkpoint by disrupting the microtubule cytoskeleton. KRIBB3 was proven to be a tubulin inhibitor using in vitro polymerization assays and in vivo indirect immunofluorescence staining. The temporal pattern of Bax activation by KRIBB3 was similar to PARP cleavage, suggesting that Bax is a mediator of KRIBB3-dependent apoptosis. Furthermore, when KRIBB3 was administered intraperitoneally into nude mice at 50 mg/kg or 100 mg/kg, it inhibited 49.5 or 70.3% of tumor growth, respectively. These results suggest that KRIBB3 is a good drug candidate for cancer therapy. PMID- 17915195 TI - Thyroxine (T4) transfer from CSF to choroid plexus and ventricular brain regions in rabbit: contributory role of P-glycoprotein and organic anion transporting polypeptides. AB - This study investigated the transfer of T4 from cerebrospinal fluid (CSF) into the choroid plexuses (CP) and ventricular brain regions, and the role of P glycoprotein (P-gp), multidrug resistance protein 1 (mrp1) and organic anion transporting polypeptides (oatps). During in vivo ventriculo-cisternal (V-C) perfusion in the anesthetized rabbit (meditomidine hydrochloride 0.5 mg kg(-1), pentobarbitone 10 mg kg(-1) i.v.), 125I-T4 was perfused continuously into ventricular CSF with reference molecules 14C-mannitol and blue dextran. Over 2 h, 36.9+/-4.6% 125I-T4 was recovered in cisternal CSF. Addition of P-gp substrate verapamil increased CSF 125I-T4 recovery to 51.4+/-2.8%, although mrp1 and oatp substrates had no significant effect. In brain, 125I-T4 showed greatest accumulation in the CP (1.52+/-0.31 ml g(-1)), followed by ventricular regions (caudate putamen, ependyma, hippocampus, 0.05-0.14 ml g(-1)). At the CP, verapamil and probenecid (but not indomethacin) significantly increased 125I-T4 accumulation, implicating a role for P-gp and oatps. Of other brain regions, all three drugs increased accumulation in caudate putamen 3-5 times, and indomethacin and probenecid increased accumulation in ependyma 4-5 times. The role of P-gp was investigated further in isolated incubated CPs using 5 microg/ml C219 anti-P-gp antibody. Both 125I-T4 and 3H-cyclosporin accumulation increased by 80%, suggesting that P-gp is functional in the CP and has a role in removal of T4. Combined with the in vivo results, these studies suggest that P-gp provides a local homeostatic mechanism, maintaining CSF T4 levels. We conclude that P-gp and oatps contribute to the transfer of 125I-T4 between the CSF, CP and brain, hence regulating 125I-T4 availability in CSF. PMID- 17915196 TI - Central administration of selective melanocortin 4 receptor antagonist HS014 prevents morphine tolerance and withdrawal hyperalgesia. AB - Major problem involved in treatment of chronic pain with morphine is the development of tolerance and dependence. Previous studies have demonstrated the participation of melanocortin (MC) system in the development of tolerance to antinociceptive effect of morphine. However, the impact of supraspinal MC4 receptors (MC4 R) modulation on this phenomenon and morphine withdrawal hyperalgesia remained unexplored. We investigated the role of central MC4 R in acute, chronic effects and withdrawal reactions of morphine using tail flick test. Acute intracerebroventricular (icv) administration of morphine (2-20 microg/rat) exhibited antinociceptive activity, which was antagonized by subeffective dose of nonselective MC R agonist NDP-MSH (0.04 ng/rat, icv), and potentiated by subeffective dose of MC4 R antagonist HS014 (0.008 ng/rat, icv). Isobolographic analysis revealed antagonistic interaction between NDP-MSH and morphine, and additive interaction between HS014 and morphine combinations. While chronic icv infusion of morphine (20 ng/microl/h) via osmotic pump for 7 days developed tolerance to its antinociceptive effect, its discontinuation produced hyperalgesia. Co-administration of HS014 (0.008 ng/rat, icv) with chronic morphine not only delayed the development of tolerance but also prevented withdrawal hyperalgesia. Furthermore, acute treatment with HS014 (0.008 and 0.04 ng/rat, icv) dose dependently attenuated the withdrawal hyperalgesia. This suggests the involvement of central MC4 R in the mechanism of development of tolerance and dependence following chronic morphine administration. We speculate that targeting this receptor may be a novel strategy to improve the effectiveness of morphine in the treatment of chronic pain. PMID- 17915197 TI - Effect of upregulated renin-angiotensin system on per2 and bmal1 gene expression in brain structures involved in blood pressure control in TGR(mREN-2)27 rats. AB - Circadian system regulates rhythms with 24 h period including those occurring in the cardiovascular system. Inverted blood pressure profile was demonstrated in hypertensive TGR(mREN-2)27 (TGR) rats with upregulated renin-angiotensin system. To depict structures involved in the generation of the inverted pattern of blood pressure in TGR rats, we analyzed daily expression of clock genes per2 and bmal1 in the brain areas involved in the regulation of the blood pressure. Heterozygous male TGR and control rats were synchronized to the light:dark cycle 12:12 and blood samples were taken in 4 h intervals within 24 h cycle. The levels of the plasma renin activity were increased in TGR rats in comparison with controls. Brain nuclei were isolated by dissection from frozen sections. The clock gene expression was determined in the hypothalamic paraventricular and dorsomedial nuclei, dorsal vagal motor nucleus, caudal ventrolateral medulla, nucleus ambiguus, area postrema, and anteroventral third ventricle. Daily pattern of per2 expression was rhythmic in most of the nuclei studied with its highest levels at the beginning of the nighttime in both groups of rats. Expression of bmal1 peaked at the beginning of the day. We found robust differences in the clock gene expression between the TGR and control rats in the area postrema. TGR rats exerted changes in the clock gene expression in the nucleus ambiguus which receives direct innervation from the area postrema. The area postrema seems to play a key role in the transmission of signals from the periphery to the CNS. PMID- 17915198 TI - Neurophysiological properties of cells filling the neonatal medial prefrontal cortex lesion cavity. AB - Removal of the medial prefrontal cortex (mPFC) of the rat during the initial 7-12 days of life results in spontaneous filling of lesion cavity that is accompanied by recovery of cognitive and motor functions. To date, it remains uncertain whether tissue filling the lesion cavity is actually supporting the functional improvement. In the present study, we examined whether spontaneous neuronal activity could be recorded in adulthood from the tissue that fills the lesion cavity. We recorded EEG and multiunit activity in adulthood from the mPFC and the motor cortex of rats that had received neonatal mPFC lesions on post-natal day 10 (P10) or their non-lesioned littermate controls. We found similarities in both the firing pattern and firing rate of cells from the filled-in region compared to that of controls, although the power associated with peak frequencies in the delta, alpha, and beta range in the EEG recorded from the filled-in region was lower compared to controls. Overall, our results suggest that the cells found in the lesion cavity have similar neurophysiological properties to those found in normal tissue and thus should be capable of at least partially supporting the observed recovery of function. PMID- 17915199 TI - An expeditious synthesis of 4-hydroxy-2E-nonenal (4-HNE), its dimethyl acetal and of related compounds. AB - The facile one step synthesis of 4-hydroxy-2E-nonenal and its dimethyl acetal via a cross-metathesis reaction between commercially available octen-3-ol and acrolein or its dimethyl acetal is reported. The method was extended to the synthesis of C6 and C12 4-hydroxy-2E-enals, their dimethyl acetal and of the 4 hydroxy-2E-nonenoic acid (4-HNA). PMID- 17915200 TI - Critical temperatures and a critical chain length in saturated diacylphosphatidylcholines: calorimetric, ultrasonic and Monte Carlo simulation study of chain-melting/ordering in aqueous lipid dispersions. AB - Chain-ordering/melting transition in a series of saturated diacylphosphatidylcholines (PCs) in aqueous dispersions have been studied experimentally (calorimetric and ultrasonic techniques) and theoretically (an Ising-like lattice model). The shape of the calorimetric curves was compared with the theoretical data and interpreted in terms of the lateral interactions and critical temperatures determined for each lipid studied. A critical chain length has been found (between 16 and 17 C-atoms per chain) which subdivides PCs into two classes with different phase behavior. In shorter lipids, the transition takes place above their critical temperatures meaning that this is an intrinsically continuous transition. In longer lipids, the transition occurs below the critical temperatures of the lipids, meaning that the transition is intrinsically discontinuous (first-order). This conclusion was supported independently by the ultrasonic relaxation sensitive to density fluctuations. Interestingly, it is this length that is the most abundant among the saturated chains in biological membranes. PMID- 17915201 TI - Analytical and clinical performance of the Ortho-Clinical Diagnostics VITROS amino-terminal pro-B type natriuretic peptide assay. AB - BACKGROUND: Measurement of amino-terminal pro-B-type natriuretic peptide (NT proBNP) is useful for evaluating patients with heart failure (HF). METHODS: We evaluated the performance of a new automated NT-proBNP assay. RESULTS: The VITROS NT-proBNP assay had mean within-run and total imprecision of 1.0% and 3.4% at NT proBNP concentrations from 67-27,500 ng/l. Acceptable linearity, functional/analytical sensitivity were demonstrated. Anticoagulant/tube types had no effect on results. Excellent sample stability and no high-dose hook were observed. High correlation between the VITROS and Elecsys methods was demonstrated (r=0.995; P<.001), with 98.3% clinical concordance. VITROS NT-proBNP concentrations were significantly higher in HF subjects than those without (1210 versus 68 ng/l; P<.001) and associated with HF symptom severity (P<.001). The VITROS assay had AUC for HF of 0.95 (P<.001), and had excellent NPV for excluding HF. CONCLUSIONS: The automated VITROS NT-proBNP assay demonstrates excellent analytical and clinical performance for evaluating the presence and severity of HF. PMID- 17915202 TI - Robust and traditional statistical methods in the establishment of immunoglobulin E target values in external quality assessment program. AB - BACKGROUND: In an EQA scheme, peer group mean is conventionally regarded as the target after removing values exceeding the mean+/-3SD. However, this computation fails if there are some outliers. METHODS: The outliers in each original data set were separately deleted by traditional and robust statistical methods. The data distributions of original and trimmed data sets were separately tested by Shapiro Wilks or Kolmogorov-Smirnov Z test. The means of peer groups were used to set IgE targets. The difference between targets set by both methods was tested by 1 sample t test. RESULTS: The original data sets were not all Gaussian distributed. After deletion of outliers using robust statistical method, the modified data sets of all tested groups were normally distributed. IgE targets established by robust and traditional methods were significantly different in some groups. The incompatible IgE targets derived all from the original and trimmed data sets which were not Gaussian distributed. CONCLUSIONS: The premise for choosing a traditional method to delete outliers and using peer group means as targets was that the data sets must be Gaussian distributed. The reasons that caused the targets to be incompatible possibly were the 2 rules to be broken. Robust statistical method was used for deletion of outliers due to the distorted distribution. PMID- 17915203 TI - Association of low repair efficiency with high hormone receptors expression and SOD activity in breast cancer patients. AB - OBJECTIVES: To evaluate the antioxidant status and repair capacity in breast cancer patients as well as the relationship between these parameters and expression of critical proteins in breast cancer tissue. DESIGN AND METHODS: Blood samples were obtained from 25 female breast cancer patients and 19 healthy women. The antioxidant status was determined by the concentration of thiobarbituric-reactive substances (TBARS) and activity of superoxide dismutase (SOD) and catalase (CAT). The basal DNA damage and repair capacity in lymphocytes were evaluated by comet assay. The expression of p53, c-erbB2, Ki-67, estrogen receptor (ER) and progesterone receptor (PR) in cancer tissue was detected by immunohistochemical staining. RESULTS: The breast cancer patients presented significantly elevated endogenous DNA damage in lymphocytes and lower susceptibility to DNA damage induced by H(2)O(2) when compared to the control group. There is a negative correlation between TBARS and sensitivity to peroxide induced DNA damage in patients. The percentage of residual damage after H(2)O(2) treatment followed by 3h of post-incubation is significantly higher in patients and also correlates positively with SOD activity, ER and PR expression and negatively with the basal DNA damage. CONCLUSIONS: Our results demonstrate low repair capacity in lymphocytes of breast cancer patients and suggest that the regulation of DNA repair is sensitive to cellular redox state and can be modulated by ER/PR status. PMID- 17915204 TI - Diagnostic accuracy of BNP and NT-proBNP in patients presenting to acute care settings with dyspnea: a systematic review. AB - OBJECTIVE: We sought to compare the diagnostic performance of B-type natriuretic peptide (BNP) and N-terminal proBNP measurements in patients presenting to acute care settings with dyspnea, a common presenting symptom of heart failure. DESIGN AND METHODS: We conducted a systematic review of the literature. For all included studies, we applied the QUADAS 14-question quality assessment tool for systematic reviews of diagnostic accuracy and abstracted the data for every published cut point. RESULTS: We screened 4338 studies and included nine in the meta-analysis. All 9 studies scored positively on at least 50% of the QUADAS questions. The pooled estimates of sensitivity and specificity were the same for the BNP studies (0.97 (95% CI: 0.96, 0.98) and 0.70 (95% CI: 0.56, 0.85)) as for the NT-proBNP studies (0.95 (95% CI: 0.90, 1.01) and 0.72 (95% CI: 0.53, 0.90)). Tests for heterogeneity were significant in both subgroups: BNP (I(2)=97.9%, p<0.001) and NT-proBNP (I(2)=87.5%, p<0.001). Similar overall results were found for the likelihood and diagnostic odds ratios. CONCLUSIONS: BNP and NT-proBNP have very similar diagnostic performance characteristics and can be used to rule out heart failure as a cause of dyspnea in the acute clinical setting. However, there is no easily identifiable optimum cut point value for each peptide. PMID- 17915205 TI - Projective filtering of time warped ECG beats. AB - The paper proposes a modification of the nonlinear state-space projections (NSSP) method. Our approach, when applied to ECG signal processing, considerably improves the method's performance. One of the crucial operations in NSSP is the search for neighborhoods of the state-space trajectory points. The modification proposed is based on imposing a few restrictions on the time location of the neighborhood points. Dynamic time warping, a technique which allows for nonlinear alignment of time series or sequences of vectors, is applied as a straightforward solution to the task of neighborhoods determination with the restrictions imposed. The influence of nonlinear alignment on the distributions of the determined neighborhoods is presented, and the resulting method of ECG enhancement is investigated. PMID- 17915206 TI - D-JNK signaling in visceral muscle cells controls the laterality of the Drosophila gut. AB - Although bilateral animals appear to have left-right (LR) symmetry from the outside, their internal organs often show directional and stereotypical LR asymmetry. The mechanisms by which the LR axis is established in vertebrates have been extensively studied. However, how each organ develops its LR asymmetric morphology with respect to the LR axis is still unclear. Here, we showed that Drosophila Jun N-terminal kinase (D-JNK) signaling is involved in the LR asymmetric looping of the anterior-midgut (AMG) in Drosophila. Mutant embryos of puckered (puc), which encodes a D-JNK phosphatase, showed random laterality of the AMG. Directional LR looping of the AMG required D-JNK signaling to be down regulated by puc in the trunk visceral mesoderm. Not only the down-regulation, but also the activation of D-JNK signaling was required for the LR asymmetric looping. We also found that the LR asymmetric cell rearrangement in the circular visceral muscle (CVM) was regulated by D-JNK signaling and required for the LR asymmetric looping of the AMG. Rac1, a Rho family small GTPase, augmented D-JNK signaling in this process. Our results also suggest that a basic mechanism for eliciting LR asymmetric gut looping may be conserved between vertebrates and invertebrates. PMID- 17915207 TI - Spatiotemporal patterns of neurogenesis in the appendicularian Oikopleura dioica. AB - Incorporation of the thymidine analog bromodeoxyuridine (BrdU) was used to assess cytogenesis in the central nervous system (CNS) of the appendicularian Oikopleura dioica. A series of timed cumulative labelings carried out from 45 minutes (min) to 8 hours (h) after fertilization provided labeling patterns that showed when neurons and support cells residing at specific sites within the 9 h CNS became postmitotic. Throughout the CNS, which includes the cerebral ganglion, caudal ganglion and caudal nerve cord, neurogenesis occurs during an earlier time window than the genesis of support cells. Neurons are first generated at about 45 min to 1 h after fertilization in all 3 CNS regions, starting in the cerebral ganglion. Support cells are generated starting at about 2 h after fertilization. In both the cerebral ganglion and the caudal ganglion, neurons born during different time epochs settle in a specific spatial pattern, following a caudal to rostral gradient in the caudal ganglion and a more complex pattern in the cerebral ganglion. No such regional pattern was seen in the caudal nerve cord, where neurons born during different epochs were evenly distributed along the length of the cord. In the cerebral ganglion a small subpopulation of cells continued to incorporate BrdU from 8 h to at least 15 h and may represent a reserve of stem cells or progenitor cells that generate additional cells seen in the adult. The results show that this simple urochordate exhibits several vertebrate features of CNS cytogenesis, including a different timing of neurogenesis and gliogenesis (support cells being the likely candidates for glial cells in Oikopleura), gradients of neuron position according to birthdate, and a maintenance of neural cell precursors beyond embryonic and larval stages. PMID- 17915208 TI - The monolayer formation of Bergmann glial cells is regulated by Notch/RBP-J signaling. AB - The Bergmann glia is a unipolar astrocyte in the cerebellar cortex, displaying a tight association with Purkinje cells. The cell bodies of Bergmann glia are located in a row around Purkinje cell somata; they extend radially arranged Bergmann fibers which enwrap the synapses on the Purkinje cell dendrites. It is well known that Bergmann glial somata migrate from the ventricular zone through the mantle zone, forming an epithelium-like lining in the Purkinje cell layer during development. However, the mechanism of the monolayer formation of Bergmann glia is poorly understood. Several reports have suggested that Notch signaling plays instructive roles in promoting the identities of several types of glial cells, including Bergmann glia. Moreover, Notch receptors are expressed in Bergmann glia during development. Here, we have deleted the Notch1, Notch2 and RBP-J genes in the Bergmann glia by GFAP-driven, Cre-mediated recombination, to study the role of Notch-RBP-J-signaling in the monolayer formation of Bergmann glia. Notch1/2- and RBP-J-conditional mutant mice showed disorganization of Bergmann fibers, irregularities of the Bergmann glial lining and aberrant localization of Bergmann glia in the molecular layer. Thus, Notch-RBP-J signaling plays crucial roles in the monolayer formation and morphogenesis of Bergmann glia. PMID- 17915209 TI - Organochlorine pesticide exposure in children living in southern Spain. AB - Despite the prohibition of most persistent organochlorine (OC) pesticides in Spain, their presence has been widely documented in adult human tissues. However, scarce information is available on the exposure of children. The aim of the present study was to investigate the presence of 16 OC pesticide residues in 52 fat samples collected from boys with a mean age of 7yr (0-15yr) living in Southern Spain and to assess the association between OC pesticide levels and child characteristics. No pesticide was found in more than 50% of samples, except for p,p'-DDE (79% of samples; median, 710ng/g lipid). After this compound, the most frequent pesticides were o,p'-DDT (17%; median, 330ng/g lipid) and o,p'-DDD (15%; median, 1510ng/g lipid). No statistically significant association was found between p,p'-DDE or SigmaDDTs and the birth year, birth weight, gestational age, infant feeding history or the age, weight, height or Quetelet Index at the time of sampling. The lack of correlation between the presence of the main metabolite p,p'-DDE and that of the parent compounds, o,p'-DDT and p,p'-DDT, suggests that children were exposed mainly to the metabolite rather than to the commercial pesticide, which was banned 30yr ago. In contrast, among currently used OCs, endosulfan was positively correlated with the presence of its metabolites, suggesting exposure to the commercial products. Further research is warranted to investigate the health consequence in children resulting from exposure to chemicals suspected of endocrine-disrupting effects. PMID- 17915210 TI - Short-term exposure to air pollution and inflammation-sensitive biomarkers. AB - OBJECTIVES: To evaluate the effect of short-term exposure to air pollutants on inflammation-sensitive biomarkers in apparently healthy individuals. METHODS: We enrolled all participants from The Tel-Aviv Sourasky Medical Center inflammation survey held between 2003 and 2006, excluding participants with an acute or chronic inflammatory disease, pregnancy, steroidal or nonsteroidal treatment, or a recent invasive procedure. Additional subjects were excluded for living more than 11km from the nearest air pollution monitoring station. Analysis was performed separately for men and women. Linear regression models were fitted for each inflammatory variable against air pollutant variables (particulate matter under 10microm, sulfur dioxide, nitrogen dioxide, carbon monoxide, and ozone) for increasing lag times of up to 7 days, and adjusted for all possible and known confounding parameters. RESULTS: The study population comprised 3659 individuals (2203 males and 1456 females). We found a statistically significant negative correlation in the male population between air pollutants, mainly NO2, SO2, and CO, and fibrinogen in several lag days. A positive correlation was found for PM10 at day 7. No such correlation was found for CRP and WBC, or for the female population. CONCLUSION: Our findings do not support the potential link between short-term exposure to air pollution and enhanced inflammation as a possible explanation for increased cardiovascular morbidity. Additional large-scale population-based studies with good methodological design are needed in order to clarify this issue. PMID- 17915211 TI - Biomarkers of manganese exposure in a population living close to a mine and mineral processing plant in Mexico. AB - Manganese (Mn) is considered an essential metal; nevertheless, excessive Mn exposure in humans is known to affect central nervous system. Mn access to its toxic target, the brain, is a complex phenomenon subject to physiological and physiopathological processes; in which, among others, the route of exposure plays an important role. Mn airborne exposure has gained interest both in occupational and environmental studies in order to understand the effects of low-level, long term exposure. The objective of the present study was to describe the relationship between blood Mn and prolactin as marker of effect exposure, as well as other variables from subjects dwelling in a mining district in central Mexico environmentally exposed to the metal. This study was conducted on 230 volunteers; blood samples were obtained from cubital vein and hemoglobin, prolactin, lead (Pb), and Mn levels were measured. Non-parametrical Spearman's correlation showed statistical associations between blood and Mn levels and prolactin (rho=0.197), hemoglobin (rho= -0.213), age (rho= -0.186), and blood lead (rho= -0.167). Multiple regression analysis showed that blood Mn levels as an important factor to determine serum prolactin levels (beta=0.111, p=0.029) in a model corrected by gender and age. Results suggest that assessment of Mn exposure by biomarkers on general population is complex due to the variability and characteristics of the metal; however, specific subpopulations such as iron-deficient individuals are suspected to accumulate Mn in blood and thus they may be susceptible to the neurotoxic effects of Mn. PMID- 17915212 TI - Environmental fate and partition co-efficient of oestrogenic compounds in sewage treatment process. AB - The presence of residual pharmaceuticals and environmental endocrine disrupters (EEDs) is increasingly significant due to their impact on human health and wildlife. Of the compounds implicated as EEDs, the most potent in their oestrogenic effect are the natural and synthetic oestrogens. As these compounds will be present in the sewage matrix, it is necessary to establish their fate during sewage treatment with a view of removal and safe disposal to avoid unnecessary exposure. Using methodology developed by the author this paper describes the results of a study undertaken to determine both the K(ow) and the adsorption characteristics of these oestrogens. The experimental values obtained were compared to a computational default model. However, there was disparity between the default model and the values determined experimentally. This was especially the case in the determination of the K(oc) which impacts directly on the sludge adsorbance potential. The calculated results ranged from log4.21 for beta-oestradiol to log4.68 for 17alpha-EE-3-ME, the experimental results were higher log(5.04-log5.83), respectively. The implications of the findings in terms of water recycling and sewage sludge disposal are also discussed. PMID- 17915213 TI - Repeated instillations of Dermatophagoides farinae into the airways can induce Th2-dependent airway hyperresponsiveness, eosinophilia and remodeling in mice: effect of intratracheal treatment of fluticasone propionate. AB - Dermatophagoides farinae are known to be a common environmental allergen causing allergic asthma; however, little is known about their pathophysiological effect via the allergenicities in vivo. Therefore, we first established a mouse model of asthma induced by repeated instillations of D. farinae. Second, to investigate whether the asthmatic responses are Th2-dependent, we examined the effect of the deficiency of interleukin-4 (IL-4) receptor alpha chain gene. Finally, we examined the effect of fluticasone propionate on this model. Mice were instilled with D. farinae without additional adjuvants into the trachea 8 times. After the final allergen instillation, the airway responsiveness to acetylcholine was measured, and bronchoalveolar lavage and histological examination were carried out. The instillation of the allergen-induced airway hyperresponsiveness, the accumulation of inflammatory cells and increases in the levels of Th2 cytokines and transforming growth factor-beta(1) production in the bronchoalveolar lavage fluid dose dependently. The number of goblet cells in the epithelium and the extent of the fibrotic area beneath the basement membrane were also increased in the morphometric study. In contrast, the defect of IL-4/IL-13 signaling through IL-4 receptor alpha chain completely abrogated all these responses. Furthermore, the simultaneous instillation of fluticasone propionate with the allergen showed significant inhibition or an inhibitory tendency of these changes. These findings demonstrate that the repetitive intratracheal instillations of D. farinae can induce airway remodeling through Th2-type inflammation, and that fluticasone propionate inhibits D. farinae-induced airway remodeling in mice, and this model would be useful for studying mechanisms involved in the development of allergic asthma. PMID- 17915215 TI - Phosphorylation, desensitization and internalization of human alpha1B adrenoceptors induced by insulin-like growth factor-I. AB - The effect of insulin-like growth factor-I (IGF-I) on human alpha(1B) adrenoceptor function, phosphorylation state and cellular location was studied. Rat-1 fibroblasts were transfected with a plasmid construction containing enhanced green fluorescent protein joined to the carboxyl terminus of the human alpha(1B)-adrenoceptor. Receptors were identified by radioligand binding and photoaffinity labeling, and were immunoprecipitated with an antiserum generated against the enhanced green fluorescent protein. The receptor was functional, as evidenced by noradrenaline action on intracellular calcium and inositol phosphate production. IGF-I had no significant effect by itself on these parameters but markedly reduced the effects of noradrenaline. IGF-I induced alpha(1B) adrenoceptor phosphorylation, which was markedly reduced by the following agents: pertussis toxin, a metalloproteinase inhibitor, diphtheria toxin mutant CRM 197, an epidermal growth factor (EGF) receptor intrinsic kinase activity inhibitor, and by phosphoinositide 3-kinase and protein kinase C inhibitors. IGF-I action appears to involve activation of a pertussis toxin-sensitive G protein, shedding of heparin-binding EGF and autocrine activation of EGF receptors. G protein subunits and phosphotyrosine residues stimulate phosphoinositide 3-kinase activity leading to activation of protein kinase C, which in turn phosphorylates alpha(1B)-adrenoceptors. Confocal fluorescent microscopy showed that alpha(1B) adrenoceptors fussed to the green fluorescent protein were located in plasma membrane and intracellular vesicles in the basal state. IGF-I induced receptor redistribution favoring the intracellular location; this effect was blocked by hypertonic sucrose and concanavalin A. Our data show that IGF-I induces alpha(1B) adrenoceptor desensitization associated to receptor phosphorylation and internalization. PMID- 17915214 TI - Acetylbritannilactone suppresses lipopolysaccharide-induced vascular smooth muscle cell inflammatory response. AB - To investigate the mechanism of action by which a new anti-inflammatory active compound, 1-O-acetylbritannilactone (ABL) isolated from Inula britannica-F., inhibits inflammatory responses in vascular smooth muscle cells (VSMCs). Enzyme immunoassay was used to measure the levels of prostandin E(2) (PGE(2)) production. Immunocytochemistry staining and Western blot analysis were performed to detect the nuclear translocation of nuclear factor-kappaB (NF-kappaB) p65 and the expression of IkappaB-alpha, pIkappaB-alpha and cyclooxygenase-2 (COX-2). Electrophoretic mobility shift assays (EMSA) were used to detect DNA-binding activity of NF-kappaB in VSMCs. ABL (5, 10, 20 micrommol/l) had several concentration-dependent effects, including inhibition of lipopolysaccharide (LPS) induced PGE(2) production and COX-2 expression, and blockade of NF-kappaB activation and translocation. These effects were owing to reductions in IkappaB alpha phosphorylation and degradation induced by LPS. In addition, ABL directly inhibited the binding of active NF-kappaB to specific DNA cis-element. These results indicate that ABL is a potent inhibitor of LPS-stimulated VSMC inflammatory responses through blockade of NF-kappaB activity and inhibition of inflammatory gene COX-2 expression. PMID- 17915216 TI - TGFbeta stimulated re-epithelialisation is regulated by CTGF and Ras/MEK/ERK signalling. AB - The complex mechanisms by which transforming growth factor beta (TGFbeta) regulate re-epithelialisation following injury of stratified epithelia are not fully understood. TGFbeta signals via binding to distinct receptors activating downstream effectors, including Smads which initiate transcription of target genes. However, studies have shown that TGFbeta can also signal independently of Smads through MAPK pathways, demonstrating the diversity of TGFbeta signalling. Connective tissue growth factor (CTGF) is strongly induced by and acts downstream of TGFbeta causing pathophysiology in tissues by inducing matrix deposition, conversion of fibroblasts into contractile myofibroblasts (e.g. dermis and corneal stroma) and stimulation of epithelial-to-mesenchymal transition (e.g. kidney and lung) all of which are known to cause fibrosis. However, a role for CTGF in epithelial cell function which does not involve direct contribution to fibrosis has not been demonstrated. We show for the first time that synthesis of CTGF in cultures of human corneal epithelial cells is induced by TGFbeta through the Ras/MEK/ERK MAPK signalling pathway and that this is required for re epithelialisation to occur through cell migration. These data reveal a novel function for CTGF in the regulation of epithelial tissue repair beyond its established role in fibrosis, and further highlight the complexity of TGFbeta regulation of epithelial cell function. PMID- 17915217 TI - Collagen gel contraction by ARPE-19 cells is mediated by a FAK-Src dependent pathway. AB - Proliferative vitreoretinopathy (PVR) may result in part from de-differentiation of retinal pigment epithelium (RPE) in an aberrant wound-healing strategy. An in vitro model of PVR, collagen gel contraction by RPE, likely requires integrin engagement and activation as an important initial step. The purpose of this study was to identify the important associated integrins and signal transduction pathway. The retinal pigment epithelial cell line ARPE-19 was used in these studies. Cell surface integrin expression was assessed using flow cytometry. An in vitro contraction assay was performed and the percent contraction quantified at specific time intervals using image capture (Gel Doc) and NIH Image software. Cells were pretreated with either small molecule inhibitors of signal transduction pathways or monoclonal antibodies with specificity for specific integrin isoforms. Transient transfections with a FAK siRNA were used to decrease FAK expression. ARPE-19 cells express alpha1, alpha2, and alpha3 integrin, isoforms involved in collagen ligation. Cell surface integrin blockade using anti integrin alpha2 (P=0.02), alpha3 (P=0.01), or a combination of alpha1, alpha2, and alpha3 (P=0.001) antibodies significantly reduced collagen gel contraction. Inhibition of the FAK-Src complex, but not MEK or PI3K, significantly decreased contraction (P=0.0001). FAK siRNA transient transfection significantly reduced FAK protein expression by 71% (P=0.02) and concordantly decreased gel contraction (P=0.0001). RPE-mediated collagen gel contraction is a multi-step process. Integrin ligation and FAK-Src activation is necessary for collagen gel contraction produced by the ARPE-19 cell line. Validation of these observations in primary RPE cells may suggest new targets for therapeutic intervention in PVR. PMID- 17915219 TI - NOD2 transgenic mice exhibit enhanced MDP-mediated down-regulation of TLR2 responses and resistance to colitis induction. AB - BACKGROUND & AIMS: Mutations in the CARD15 gene encoding NOD2 are susceptibility factors in Crohn's disease. We explored the mechanism of this susceptibility using mice that over express NOD2. METHODS: Cellular and molecular responses of mice bearing an NOD2 transgene or administered plasmids that express wild-type and mutated NOD2 constructs were examined. RESULTS: In initial studies, we showed that splenocytes from NOD2 transgenic mice as compared with littermate controls exhibit decreased interleukin (IL)-12p70 responses to peptidoglycan (PGN), a TLR2 ligand that contains muramyl dipeptide, but not other TLR ligands; in contrast, IL-12 responses to PAM(3)CSK(4), a TLR2 ligand that does not contain muramyl dipeptide, were normal. Similarly, transgenic mice as compared with controls exhibited greatly decreased IL-12p40 responses to intraperitoneal administration of PGN but not to lipopolysaccharide. In further studies, we showed using electrophoretic mobility shift assay that PGN-stimulated cells from transgenic mice exhibited decreased activation of nuclear factor kappaB. Finally, in a series of studies on the effect of the NOD2 on susceptibility to induced colitis, we found that (1) transgenic mice were highly resistant to induction of PGN colitis and partially resistant to induction of trinitrobenzene sulfonic acid (TNBS) colitis and (2) mice administered a plasmid expressing a wild-type NOD2 gene were completely resistant to TNBS colitis whereas mice administered a plasmid expressing an NOD2 gene with the Crohn's disease frameshift mutation were only slightly resistant to TNBS colitis. CONCLUSIONS: These data offer new evidence that NOD2 mutations contribute to inflammatory bowel disease by causing excessive TLR2 cytokine responses. PMID- 17915218 TI - VEGF modulation of retinal pigment epithelium resistance. AB - Fluid accumulation into the subretinal space and the development of macular edema is a common condition in age-related macular degeneration, diabetic retinopathy, and following ocular surgery, or injury. Vascular endothelial growth factor (VEGF) and other cytokines have been implicated in the disruption of retinal pigment epithelium (RPE) barrier function and a reduction in the regulated removal of subretinal fluid; however, the cellular and molecular events linking these agents to the disruption of barrier function have not been established. In the current study, cultures of ARPE-19 and primary porcine retinal pigment epithelium (RPE) cells were utilized to investigate the effects of the VEGF induced modifications to the barrier properties of the RPE. The barrier function was determined by transepithelial resistance (TER) measurements and morphology of the RPE monolayers. In both ARPE-19 and primary porcine RPE cells the administration of VEGF produced a significant drop in TER, and this response was only observed following apical administration. Maximum reduction in TER was reached 5h post VEGF administration. These responses were concentration-dependent with an EC(50) of 502pg/mL in ARPE-19 cells and 251pg/mL in primary porcine cells. In both ARPE-19 and primary RPE cells, the response to VEGF was blocked by pretreatment with the relatively selective VEGF-R2 antagonists, SU5416 or ZM323881, or the protein tyrosine kinase inhibitor, genistein. Administration of the relatively selective VEGF-R2 agonist, VEGF-E, also reduced TER in a concentration-dependent manner (EC(50) of 474pg/mL), while VEGF-R1 agonist, placental growth factor (PlGF), did not significantly alter the TER. Immunolocalization studies demonstrated that confluent monolayers exhibited continuous cell-to-cell ZO-1 protein contacts and apical localization of the VEGF R2 receptors. These data provide evidence that the VEGF-induced breakdown of RPE barrier function is mediated by the activation of apically-oriented VEGF-R2 receptors. Thus, VEGF-mediated increases in RPE permeability are initiated by a rise in intraocular levels of VEGF. PMID- 17915220 TI - Pre-S deletion and complex mutations of hepatitis B virus related to advanced liver disease in HBeAg-negative patients. AB - BACKGROUND & AIMS: This longitudinal study investigated the interactions and roles of hepatitis B virus (HBV) genotypes, pre-S deletions, and core promoter and precore mutations on the progression of liver disease in hepatitis B e antigen (HBeAg)-negative patients. METHODS: A total of 141 HBeAg-negative patients without liver cirrhosis or hepatocellular carcinoma at study entry were recruited for this study, including 45 inactive HBV carriers and 96 patients with HBeAg-negative chronic hepatitis B. The HBV genotypes and the sequences of pre-S, core promoter, and precore regions were determined. RESULTS: Compared with patients without developing liver cirrhosis, patients with the development of liver cirrhosis had higher rates of genotype C; pre-S deletions; C or G1753, T1762/A1764, T1766, and/or A1768 mutants; and G1799 variant. Cox regression analysis showed that older age, higher total bilirubin and HBV DNA levels, pre-S deletions, and T1766 and/or A1768 mutants were significantly associated with the development of liver cirrhosis. HBV with a complex mutation pattern (pre-S deletion, T1762/A1764, and T1766 and/or A1768 mutants) rather than a single mutation was associated with the development of liver cirrhosis, and the patterns of mutation combinations differed between HBV genotype B and C. Moreover, pre-S deletion was a significant risk factor for hepatocellular carcinoma. CONCLUSIONS: This study indicated that pre-S deletion and combined mutations of HBV are useful molecular markers for predicting the clinical outcomes of HBeAg-negative patients. PMID- 17915221 TI - TIM-4 expressed by mucosal dendritic cells plays a critical role in food antigen specific Th2 differentiation and intestinal allergy. AB - BACKGROUND & AIMS: Food allergy accounts for significant morbidity. The etiology and immune mechanisms of food allergy, however, have remained poorly understood. In this study, we aimed to determine the role of T-cell immunoglobulin-domain and mucin-domain (TIM)-4, a recently identified member of cell surface molecules, in the pathogenesis of intestinal allergy in a murine model. METHODS: We report that TIM-4 as well as costimulatory molecules were up-regulated in intestinal mucosal dendritic cells by in vitro or in vivo exposure to Staphylococcus enterotoxin B (SEB). SEB-conditioned intestinal dendritic cells loaded with a food macromolecule ovalbumin (OVA) induced potent OVA-specific T-helper (Th)2 lymphocyte responses in vitro and such Th2 responses were inhibited completely by TIM-4 blockade. RESULTS: In vivo exposure to both SEB and OVA resulted in OVA specific Th2 differentiation and intestinal allergic responses including increased serum immunoglobulin E and Th2 cytokine levels, activation of OVA specific Th2 cells detected both ex vivo and in situ, and mast cell degranulation. Of importance, in vivo abrogation of TIM-4 or its cognate ligand TIM-1 by using a polyclonal antibody remarkably dampened Th2 differentiation and intestinal allergy. CONCLUSIONS: Our study thus identifies TIM-4 as a novel molecule critically required for the development of intestinal allergy. PMID- 17915222 TI - Does testosterone mediate the trade-off between nestling begging and growth in the canary (Serinus canaria)? AB - Nestling birds solicit food from their parents with vigorous begging displays, involving posturing, jostling and calling. In some species, such as canaries, begging is especially costly because it causes a trade off against nestling growth. Fitness costs of begging like this are predicted by evolutionary theory because they function to resolve conflicts of interest within the family over the provision of parental investment. However, the mechanism that links these costs with nestling behaviour remains unclear. In the present study, we determine if the relationships between nestling androgen levels, nestling begging intensities and nestling growth rates are consistent with the hypothesis that testosterone is responsible for the trade-off between begging and growth. We test this idea with a correlational study, using fecal androgens as a non-invasive method for assaying nestling androgen levels. Our results show that fecal androgen levels are positively correlated with nestling begging intensity, and reveal marked family differences in each trait. Furthermore, changes in fecal androgen levels between 5 and 8 days after hatching are positively associated with changes in nestling begging intensity, and negatively associated with nestling growth during this time. Although these correlational results support our predictions, we suggest that that experimental manipulations are now required to test the direct or indirect role of testosterone in mediating the trade-off between begging and growth. PMID- 17915223 TI - The influence of personal characteristics on the resilience of operating room nurses: a predictor study. AB - BACKGROUND: Resilience in the workplace has been described as a means of facilitating adaptation in stressful environments, and therefore has application in nursing contexts. However, little research has examined how personal characteristics such as age, nursing experience and education contribute to resilience in clinical environments such as the operating room (OR). OBJECTIVE: First to identify the level of resilience, and second, investigate whether age, experience and education contribute to resilience in an Australian sample of OR nurses. METHODS: A predictive survey design was used. A random sample of 1430 nurses who were members of the Australian College of Operating Room Nurses association were surveyed. The survey included the 25-item Connor-Davidson Resilience Scale, and demographic questions. A standard regression model tested the hypothesis that age, years of OR experience and education contributed to resilience in OR nurses. RESULTS: A total of 735 (51.4%) completed, usable surveys were returned. Pearson's correlations demonstrated modest but statistically significant associations between age (p<0.001), and years of OR experience (p<0.0001), and resilience. In the multiple regression model, only years of OR experience predicted resilience (p<0.0001) and explained a small 3.1% of the variance in resilience. CONCLUSIONS: In OR nurses, resilience appears to be predicted by other attributes and is not necessarily dependent on an individual's personal characteristics. Thus, recruitment to the OR should not be based on the conventional notion that an older nursing workforce will have greater longevity and hence be more stable. If younger, less experienced nurses are adequately supported, they may thrive in the OR environment. PMID- 17915224 TI - The role of hydrostatic pressure in foam cell formation upon exposure of macrophages to LDL and oxidized LDL. AB - Hypertension is a major, established risk factor for atherosclerosis. How it interacts to exacerbate the cellular processes involved in atherogenesis is unclear. This initial, preliminary study examined how hydrostatic pressure influenced the formation of foam cells from human macrophages exposed to low density lipoprotein (LDL) or oxidized LDL (OxLDL). The results demonstrated that both LDL and OxLDL, at physiological concentration, were taken up by cultured human macrophages and foam cells were formed. This led to cell detachment and death within 24h. These effects were more rapid and more pronounced in pressurized cultures. We conclude that exposure of cell cultures to cyclical hydrostatic pressure (CHP) aggravated the adverse effects of the lipids on the macrophages. PMID- 17915225 TI - Validation of CFD predictions of flow in a 3D alveolated bend with experimental data. AB - Verifying numerical predictions with experimental data is an important aspect of any modeling studies. In the case of the lung, the absence of direct in vivo flow measurements makes such verification almost impossible. We performed computational fluid dynamics (CFD) simulations in a 3D scaled-up model of an alveolated bend with rigid walls that incorporated essential geometrical characteristics of human alveolar structures and compared numerical predictions with experimental flow measurements made in the same model by particle image velocimetry (PIV). Flow in both models was representative of acinar flow during normal breathing (0.82ml/s). The experimental model was built in silicone and silicone oil was used as the carrier fluid. Flow measurements were obtained by an ensemble averaging procedure. CFD simulation was performed with STAR-CCM+ (CD Adapco) using a polyhedral unstructured mesh. Velocity profiles in the central duct were parabolic and no bulk convection existed between the central duct and the alveoli. Velocities inside the alveoli were approximately 2 orders of magnitude smaller than the mean velocity in the central duct. CFD data agreed well with those obtained by PIV. In the central duct, data agreed within 1%. The maximum simulated velocity along the centerline of the model was 0.5% larger than measured experimentally. In the alveolar cavities, data agreed within 15% on average. This suggests that CFD techniques can satisfactorily predict acinar-type flow. Such a validation ensure a great degree of confidence in the accuracy of predictions made in more complex models of the alveolar region of the lung using similar CFD techniques. PMID- 17915226 TI - Direct parameterization of postural stability during quiet upright stance: effects of age and altered sensory conditions. AB - The purpose of this study was to determine the relationship between measures of local dynamic stability (LDS) during upright stance and both descriptive measures of postural sway and a scaling index (alpha) derived from detrended fluctuation analysis. Center of pressure (COP) time series were obtained from healthy participants (16 young and 16 older) during upright quiet stance. Vision and somatosensation were altered by eye closure and standing on a compliant surface, respectively. A non-linear time-series analysis method was used to compute three LDS parameters from the COP data: A which was defined as the COP excursion range in state space, and tau(S) and tau(L) which were defined as the divergence rates over short- and long-term timescales, respectively. LDS parameters, descriptive COP measures, and alpha had generally consistent sensitivities to age and/or altered sensory conditions. Age x sensory condition interactions, however, had distinct effects on LDS parameters compared to the other COP-based measures. Older individuals exhibited faster divergence rates while having similar magnitudes of A, compared to young individuals. These results suggest that older individuals stiffen the musculoskeletal system via increased muscle activity, perhaps as an age-related postural adaptation. In addition, correlations between LDS parameters and other COP measures were relatively small (r2<<0.29). Hence, LDS parameters (A, tau(S) and tau(L)) provide distinct information on postural control and stability, supplementing other COP-based measures. PMID- 17915227 TI - Determination of vertebral endplate deformation under load using micro-computed tomography. AB - Endplate strength plays an important role in preventing vertebral failure of normal vertebrae and in cases where surgical intervention has replaced the disc with an implant or has altered the vertebral loading. We have developed a non contact method based on the principles of image guided failure analysis, mechanical testing, and micro-computed tomography analysis, which allows for in vitro quantification of endplate deformation under axial load. The method allows for the implementation of a repeated measures experimental design, each specimen acting as its own control. Our methodology was validated using cadaveric functional spine units, loaded stepwise from 200 N to a maximum of 2000 N. The loading protocol was repeated over two days, allowing time for recovery of the disc mechanical properties. We found no meaningful difference in measured force, stiffness, and endplate deformation between day 1 and day 2. The mean fiducial registration error was less than 0.015 mm for all three axes. Endplate deformation could be reproducibly estimated. The root mean squared error was 0.03 mm, which is the effective precision of the method. Using this micro-CT based method, the effect of interbody implants, grafts, disc replacement strategies, and surgical procedures such as nucleotomy and vertebral cement augmentation on endplate mechanical behaviour can be ascertained. PMID- 17915228 TI - Preparation and characterization of six calixarene bonded stationary phases for high performance liquid chromatography. AB - Six calixarene bonded silica gel stationary phases were prepared and characterized by elemental analysis, infrared spectroscopy and thermal analysis. Their chromatographic performance was investigated by using PAHs, aromatic positional isomers and E- and Z-ethyl 3-(4-acetylphenyl) acrylate isomers as probes. Separation mechanism based on the different interactions between calixarenes and analytes were discussed. The chromatographic behaviors of those analytes on the calixarene columns were influenced by the supramolecular interaction including pi-pi interaction, space steric hindrance and hydrogen bonding interaction between calixarenes and analytes. Notably, the presence of polar groups (-OH, -NO(2) and -NH(2)) in the aromatic isomers could improve their separation selectivity on calixarene phase columns. The results from quantum chemistry calculation using DFT-B3LYP/STO-3G* base group were consistent with the retention behaviors of PHAs on calix[4]arene column. PMID- 17915229 TI - Anion-exchange chromatography-electrospray ionization mass spectrometry method development for the environmental analysis of aliphatic polyhydroxy carboxylic acids. AB - For environmental analytical purposes, an anion-exchange chromatography electrospray ionization MS method was developed enabling the identification and quantification of 18 aliphatic (poly)hydroxy (poly)carboxylic acids including sugar and uronic acids. The initially incomplete separation could be partially improved for specific analyte combinations adding borate, the zwitter ionic MES, or an alkylglycoside to the NaOH gradient supplied with the Dionex IonPac AS-11 HC column. Despite the incomplete separation, reproducible and sensitive identification and quantification is possible with MS detection in the selected ion-monitoring mode. Applying an internal standard for the compensation of the sensitivity drift of the detector, linear calibration functions were calculated for all analytes. Limits of detection spanned between 18 and 71 microgL(-1) which could be further improved for some more hydrophobic analytes by post-column addition of MeOH. PMID- 17915230 TI - Stereoisomer analysis of wastewater-derived beta-blockers, selective serotonin re uptake inhibitors, and salbutamol by high-performance liquid chromatography tandem mass spectrometry. AB - A reversed-phase enantioselective liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) method was developed to measure enantiomer fractions (EF) and concentrations of pharmaceuticals in wastewater. Enantiomer resolution of six beta-blockers (atenolol, metoprolol, nadolol, pindolol, propranolol, and sotalol) along with two selective serotonin re-uptake inhibitors (citalopram, fluoxetine) and one beta(2)-agonist (salbutamol) was achieved with the Chirobiotic V stationary phase. Analyte recovery averaged 86% in influent and 78% in effluent with limits of detection ranging from 0.2 to 7.5 ng/L. These results represent an improvement in wastewater EF measurement for atenolol, metoprolol and propranolol as well as the first EF measurements of citalopram, fluoxetine, nadolol, pindolol, salbutamol and sotalol in wastewaters. Changes in EF through treatment indicate biologically mediated stereoselective processes were likely occurring during wastewater treatment. PMID- 17915231 TI - Determination of betaine, choline and trimethylamine in feed additive by ion exchange liquid chromatography/non-suppressed conductivity detection. AB - An ion chromatography method with non-suppressed conductivity detection was developed for the simultaneous determination of betaine, choline and trimethylamine in feed additive. The analytes and the common inorganic cations were well separated by means of cation-exchange chromatography using a 4.5 mmol/L methanesulfonic acid solution containing 10% (v/v) acetonitrile as eluent and an IonPac SCS1 column (250 mm x 4 mm i.d.) as the separation column. The effects of the different chromatographic parameters on the separation were also investigated. Detection limits of betaine, choline and trimethylamine were 0.076, 0.044 and 0.041 mg/L. The relative standard deviations (RSDs) of the retention time and peak area were less than 0.30% and 0.88%, respectively. The recoveries were between 93.2% and 112.6%. The method is suitable for use as a routine method in production quality control of feed additive. PMID- 17915232 TI - Simultaneous determination of pesticides, polycyclic aromatic hydrocarbons and polychlorinated biphenyls in seawater and interstitial marine water samples, using stir bar sorptive extraction-thermal desorption-gas chromatography-mass spectrometry. AB - A method for the simultaneous determination of semi-volatile organic contaminants (polycyclic aromatic hydrocarbons, polychlorinated biphenyls, organochlorine and organophosphorus pesticides) in marine samples has been developed, for the first time, using the stir bar sorptive extraction technique (SBSE) and thermal desorption coupled to capillary gas chromatography-mass spectrometry (SBSE-TD-GC MS). Polydimethyl siloxane (PDMS) was used for the extraction of the selected analytes and two procedures have been optimised and validated, one for seawater samples (100 mL) and another for interstitial water samples (10 mL), using PDMS stir bars of 20 mm and 10 mm size, respectively. The extraction and analytical conditions, such as extraction time, matrix effects, sample volume and desorption time, were optimised. The proposed methods are sensitive, simple and show good linearity and detection limits lower than 1 ng L(-1) with seawater and lower than 10 ng L(-1) with interstitial marine water for the majority of compounds tested. Repeatability and reproducibility, expressed as relative standard deviation, have values lower than 20% for the majority of analytes considered. The recoveries for both sample volume procedures are higher than 60 and 70% for 10 and 100 mL, respectively, except for the more apolar (some PAHs and PCBs) and the more polar (some triazines) analytes which present lower values. The present SBSE/GC/MS method was applied for the analysis of trace organic contaminants in seawater and interstitial water samples from Cadiz Bay (SW of Spain). Terbutylazine, DDX and some PAHs were found at several seawater sampling points at ng L(-1) levels, and some PAHs in interstitial water too. PMID- 17915233 TI - Gas chromatography-olfactometry in food flavour analysis. AB - The application of gas chromatography-olfactometry (GC-O) in food flavour analysis represents to be a valuable technique to characterise odour-active, as well as character impact compounds, responsible for the characterizing odour of a food sample. The present article briefly reviews the use of GC-O in the flavour investigation of dairy products (milk and cheese), coffee, meat and fruits. Particular attention has been devoted to extraction techniques, GC-O hardware commonly utilised and olfactometric assessment methods, which can be applied to food analysis. PMID- 17915234 TI - Metabolic fingerprinting investigation of Artemisia annua L. in different stages of development by gas chromatography and gas chromatography-mass spectrometry. AB - Artemisia annua L. is an annual herb native of Asia and this plant has been famous for the discovery of the anti-malarial drug artemisinin since 1971. In this work, to investigate variety of whole metabolites, metabolic fingerprinting analysis of A. annua L. was carried out by GC and GC-MS coupled with trimethylsilyl derivatisation. Principal component analysis and partial least squares discriminant analysis were employed to classify GC data of A. annua L. samples at five developmental stages. The results indicated that there was no distinct difference of metabolites between control (001) and transgenic strain (F4) from the tender seedling stage to adult seedling stage, but clear differences were detected at pre-flower budding stage, flower budding stage and full flowering stage. Three precursors of artemisinin biosynthesis were studied at five developmental stages and found that a possible bottleneck exists in the conversion from artemisinic acid or dihydroartemisinic acid to artemisinin. PMID- 17915235 TI - Pulsed flow modulation comprehensive two-dimensional gas chromatography. AB - Pulsed flow modulation (PFM) is based on higher flow rate time compression of the first GC column effluent, which prior to the injection into the second column is stored for a few seconds in a standard fused silica wide bore transfer line. We constructed the PFM device with two standard 1/16 in. brass compression fittings with the insertion of the two columns inside the wide bore 0.53 mm i.d. fused silica storage transfer line for the elimination of dead volumes. This simple arrangement provides a combination of flexibility in the length of the sample storage transfer line hence comprehensive two-dimensional gas chromatography (GC x GC) cycle time, inert sample path and full elimination of cooling gas consumption. A record short second column injection time of 20 ms is demonstrated. Practical injection times are the sample collection time (such as 4s) divided by the second to first column flow rate ratio (such as 20/0.7), which is typically around 150 ms. Due to the low cost of the device it can also be considered for use with non comprehensive time segmented GC x GC to remove a few accidental coelutions. PFM-GCxGC excels with high second column capacity due to the use of 0.32 mm i.d. columns with high flow rates as the second dimension GC x GC column. As a result, PFM-GCxGC can have up to two orders of magnitude higher second column sample capacity and linear dynamic range for improved reduction of adverse matrix interference effects due to column overloading. PMID- 17915236 TI - Perfusion reversed-phase high-performance liquid chromatography/mass spectrometry analysis of intact soybean proteins for the characterization of soybean cultivars. AB - Perfusion reversed-phase HPLC (RP-HPLC)-electrospray mass spectrometry (ESI-MS) was employed for the characterization of soybean cultivars through the analysis of intact soybean proteins. The similarities and differences between yellow soybeans (the most usual soybeans) and other beans with different pigmentation (green, red, and black) commercialized as soybean were investigated. Red beans commercialized as azuki that are frequently sold as red soybean were also analyzed. Separation was carried out using a perfusion column at a flow-rate of 0.5 mL/min and a gradient elution. A step-by-step procedure was used for the optimization of the mass spectrometry parameters enabling the most sensitive detection. The method was applied to the analysis of the above-mentioned beans and the main soybean proteins (11S and 7S globulins) obtained by a fractionation procedure. MS spectra obtained from every peak in the beans and in their fractions were compared observing clear differences between yellow soybeans and the other beans with different pigmentation. The identification of some soybean proteins in yellow soybeans was also possible. PMID- 17915237 TI - Photo-ionisation mass spectrometry as detection method for gas chromatography. Optical selectivity and multidimensional comprehensive separations. AB - Mass spectrometry (MS) with soft ionisation techniques (i.e. ionisation without fragmentation of the analyte molecules) for gaseous samples exhibits interesting analytical properties for direct analysis applications (i.e. direct inlet mass spectrometric on-line monitoring) as well as mass spectrometric detection method for gas chromatography (GC-MS). Commonly either chemical ionisation (CI) or field ionisation (FI) is applied as soft ionisation technology for GC-MS. An interesting alternative to the CI and FI technologies methods are photo ionisation (PI) methods. PI overcomes some of the limitations of CI and FI and furthermore add some unique analytical properties. The resonance enhanced multi photon ionisation (REMPI) method uses intense UV-laser pulses (wavelength range approximately 350-193 nm) for highly selective, sensitive and soft ionisation of predominately aromatic compounds. The single photon ionisation (SPI) method utilises VUV light (from lamps or laser sources, wavelengths range approximately 150-110 nm) can be used for a universal soft ionisation of organic molecules. In this article the historical development as well as the current status and concepts of gas chromatography hyphenated to photo-ionisation mass spectrometry are reviewed. PMID- 17915238 TI - Effects of inner diameter of monolithic column on separation of proteins in capillary-liquid chromatography. AB - Polymer monolithic columns with I.D. between 100 and 320 microm were prepared by in-situ polymerization of styrene and divinylbenzene in fused silica capillaries. The effects of monolithic column I.D. on the separation of proteins in reversed phase capillary-liquid chromatography under gradient elution were systemically studied. The loading capacity was positively proportional to the volume of the stationary phase. It was found that the smaller diameter columns showed better performance for protein separation. The minimum plate height decreases from 34.99 microm (320 microm I.D. column) to 5.39 microm (100 microm I.D. column) for a retained protein. After studying the three parameters of the Van Deemter equation, it was interpreted that the smaller diameter can provide less flow resistance and the better performance may also be improved by the increasing of the effective diffusion. This conclusion was also supported by the data of separation permeability and breakthrough curves. PMID- 17915239 TI - On-line preconcentration and chiral separation of propiconazole by cyclodextrin modified micellar electrokinetic chromatography. AB - A method for the chiral separation of propiconazole using cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) with hydroxypropyl-gamma cyclodextrin (HP-gamma-CD) as chiral selector is reported. The use of a mixture of 30 mM HP-gamma-CD, 50mM SDS, methanol-acetonitrile 10%:5% (v/v) in 25 mM phosphate buffer solution was able to separate two enantiomeric pairs of propiconazole. Stacking- and sweeping-CD-MEKC under neutral pH (pH 7) and under acidic condition (pH 3.0) were used as two on-line preconcentration methods to increase detection sensitivity of propiconazole. Good repeatabilities in the migration time, peak area and peak height were obtained in terms of relative standard deviation (RSD). A sensitivity enhancement factor of 100-fold was achieved using sweeping-CD-MEKC at acidic pH. This is the first report on the separation of two pairs of propiconazole enantiomers and all the enantiomers of fenbuconazole and tebuconazole using sweeping-CD-MEKC. The limit of detection (S/N=3) for the three triazole fungicides ranged from 0.09 to 0.1 microg/mL, which is well below the maximum residue limits (MRL) set by Codex Alimentarius Commission (CAC). Combination of solid-phase extraction (SPE) pretreatment and sweeping-CD-MEKC procedure was applied to the determination of selected triazole fungicides in grapes samples spiked at concentration 10-40 times lower than the MRL established by the CAC. The average recoveries of the selected fungicides in spiked grapes samples were good, ranging from 73% to 109% with RSD of 9-12% (n=3). PMID- 17915240 TI - Measurement of electroosmotic flow in capillary and microchip electrophoresis. AB - Microfluidics is the science and technology of systems that process or manipulate small amounts of fluids, using channels with dimensions of tens of micrometers. Electroosmotic flow (EOF) is an important characteristic of fluids in microchannels. In this paper, EOF generation, effects on separation and definition of EOF are introduced. And EOF measurement methods on capillary electrophoresis (CE) and microchip CE are systematically reviewed based on detection principle, hallmarks of EOF measurement methods are presented, the devices and signals are also schematically described. This paper offers researchers a guidance to obtain an estimate of EOF mobility in capillary and microchip electrophoresis. PMID- 17915241 TI - Development of an integrated direct-contacting optical-fiber microchip with light emitting diode-induced fluorescence detection. AB - In this paper, one poly(dimethylsiloxane) (PDMS) sandwich microchip integrated with one direct-contacting optical fiber was fabricated by using a thin-casting method. This novel integrated PDMS sandwich microchip included top glass plate, PDMS membrane replica with microfluidic networks and optical fiber, flat PDMS membrane and bottom glass plate. As the tip of excitation optical fiber completely contacted with the separation microchannel in this integrated microchip, it not only increased the excitation light intensity to achieve the high sensitivity, but also reduced the diameter of excitation beam to obtain high resolution. In addition, we found that this rigid PDMS sandwich microchip structure effectively prevented PDMS microchannel distortion from rigid optical fiber, and provided a substantial convenience for microchips manipulating. A blue light-emitting diode (LED) was applied as excitation source by using optical fiber to couple excitation light into its direct-contacting microchannel for fluorescence detection. The performances of this integrated PDMS sandwich microchip was demonstrated by separating the mixture of sodium fluorescein (SF) and fluorescein isothiocyanate isomer I (FITC), and showed a higher sensitive and resolution than those obtained from the conventional integrated optical-fiber PDMS microchip with a 100-microm distance between fiber tip and separation microchannel. Additionally, the reproducibility of this integrated microchip with LED-induced fluorescence detection was also examined by separation of a mixture of FITC-labeled amino acids. PMID- 17915242 TI - Metal patterning on silicon surface by site-selective electroless deposition through colloidal crystal templating. AB - Site-selective Cu deposition on a Si substrate was achieved by a combination of colloidal crystal templating, hydrophobic treatment, and electroless plating. Uniformly sized nano/microstructures were produced on the substrate using a monolayer coating of colloidal spheres instead of a conventional resist. The Cu patterns obtained were of two different types: networklike honeycomb and isolated island patterns with a minimum period of 200 nm. Each ordered pattern with the desired intervals was composed of clusters of Cu nanoparticles with a size range of 50-100 nm. By the present method, it is possible to control the periodicity of metal arrays by changing the diameter of the colloidal spheres used as an initial mask and to adjust the shape of the metal patterns by changing the mask structure for electroless plating. PMID- 17915243 TI - Self-assembly of beta-casein and lysozyme. AB - The self-assembly of beta-casein and lysozyme, a linear and a globular protein with isoelectric point of pH 5.0 and 10.7, respectively, was studied. Polydisperse electrostatic complex micelles formed when mixing beta-casein and lysozyme aqueous solutions. After the micelle solution was heated, lysozyme gelated and beta-casein was trapped in the gel, producing narrowly dispersed nanoparticles. The nanoparticles were characterized with laser light scattering, zeta-potential, steady state fluorescence, atomic force microscopy, and transmission electron microscopy. The nanoparticles have spherical shape and their sizes depend on the pH of the heat treatment and the molar ratio of beta casein to lysozyme. The nanoparticles display amphoteric property and are relatively hydrophobic at pH around 5 and around 10. The net charges on the surface stabilize the nanoparticles in the solution. PMID- 17915244 TI - Facile and controllable synthesis of polyoxometalate nanorods within polyelectrolyte matrix. AB - Well dispersed polyoxometalate nanorods have been selectively and controllably synthesized within the polyelectrolyte (PE) films via a layer-by-layer (LbL) adsorption-precipitation method. The PE matrix was fabricated by LbL self assembly technology and then the multilayer films containing polyoxometalate nanorods were constructed by repetitive adsorption of polyanions and subsequent precipitation with counter ions-tetraethylammonium bromide (TEAB). Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) was used to observe the variation of size and morphology of the nanorods. The growth process and composition of the multilayer films containing nanorods were also studied. PMID- 17915245 TI - Composite hybrid membrane of chitosan-silica in pervaporation separation of MeOH/DMC mixtures. AB - Chitosan-silica hybrid membranes (CSHMs) were prepared by cross-linking chitosan (CS) with 3-aminopropyl-triethoxysilane (APTEOS). The dynamic behaviors of the CS membrane and the CSHM were investigated in pervaporation (PV) of methanol/dimethyl carbonate (MeOH/DMC) mixtures. The membranes were characterized by X-ray diffraction (XRD), contact angle meter, scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). The transition state of PV processes were studied. During the PV processes, the amorphous region of the membranes increases and the contact angle between MeOH and the membrane decreases within a range of operating time and then remains almost constant implying a reconstruction occurred on the membrane surface. The silica is well distributed in the CSHM matrix and the thermal stability of the CSHM is enhanced. The time for a PV process to reach a steady state decreases with increasing MeOH concentration or feed temperature, and it is longer for the CSHM than the CS membrane under the same operating condition. Swelling experiments show that the degree of swelling (DS) is greatly depressed by cross-linking CS with APTEOS. Sorption data indicate that the selectivity of solubility and diffusion of the CSHM are greatly improved over the CS membrane. The CSHM presents superior separation behaviors over other membranes with a flux of 1265 g/(hm(2)) and separation factor of 30.1 in PV separation of 70 wt% MeOH in feed at 50 degrees C. PMID- 17915246 TI - Electric-field-enhanced transport in polyacrylamide hydrogel nanocomposites. AB - Electroosmotic pumping through uncharged hydrogels can be achieved by embedding the polymer network with charged colloidal inclusions. Matos et al. [M.A. Matos, L.R. White, R.D. Tilton, J. Colloid Interface Sci. 300 (2006) 429-436], recently used the concept to enhance the diffusion-limited flux of uncharged molecules across polyacrylamide hydrogel membranes for the purpose of improving the performance of biosensors. This paper seeks to link their reported macroscale diagnostics to physicochemical characteristics of the composite microstructure. The experiments are characterized by a Debye screening length that is much larger than the radius of the silica nanoinclusions and the Brinkman screening length of the polymer skeleton. Accordingly, closed-form expressions for the incremental pore mobility are derived, and these are evaluated by comparison with numerically exact solutions of the full electrokinetic model. A mathematical model for the bulk electroosmotically enhanced tracer flux is proposed, which is combined with the electrokinetic model to ascertain the electroosmotic pumping velocity from measured flux enhancements. Because the experiments are performed with a known current density, but unknown bulk conductivity and electric field strength, theoretical estimates of the bulk electrical conductivity are adopted. These account for nanoparticle polarization, added counterions, and non-specific adsorption. Theoretical predictions of the flux enhancement, achieved without any fitting parameters, are within a factor of two of the experiments. Alternatively, if the Brinkman screening length of the polymer skeleton is treated as a fitting parameter, then the best-fit values are bounded by the range 0.9-1.6 nm, depending on the inclusion size and volume fraction. Independent pressure-driven flow experiments reported in the literature for polyacrylamide gels without inclusions suggest 0.4 or 0.8 nm. The comparison can be improved by allowing for hindered ion migration, while uncertainties regarding the inclusion surface charge are demonstrated to have a negligible influence on the electroosmotic flow. Finally, and perhaps most importantly, anomalous variations in the flux enhancement with particle size and volume fraction can be rationalized at present only by acknowledging that particle-particle and particle-polymer interactions increase the effective permeability of the hydrogel skeleton. This bears similarities to the increase in polymer free volume that accompanies the addition of silica nanoparticles to certain polymeric membranes. PMID- 17915247 TI - Shear-induced capping of L-selectin on the neutrophil surface during centrifugation. AB - L-selectin on leukocytes is critical in leukocyte tethering and adhesion to inflamed endothelium and lymphocyte homing to lymphoid organs. The spatial distribution of L-selectin on leukocytes controls cellular adhesive function in hydrodynamic shear. How L-selectin changes its position on the cell membrane remains an open question, but a possible candidate is shear stress encountered on the cell surface. Here we demonstrate shear-induced L-selectin polarization on the membrane during the process of centrifugation of resting neutrophils via immunofluorescent microscopy. It was found that randomly distributed L-selectin on neutrophils moves to a polar cap at one end of the cell after centrifugation (300 x g for 2 min) without inflammatory stimuli. This L-selectin redistribution under shear was predicted by Monte Carlo simulations that show how convection dominates over diffusion, leading to L-selectin cap formation during centrifugation at 280 x g or during leukocyte adhesion to the endothelial wall at 1 dyn/cm(2). Those results point to a role for shear stress in the modulation of L-selectin distribution, and suggest a possible alternate mechanism and reinterpretation of previous in vitro studies of L-selectin mediated adhesion of neutrophils isolated via centrifugation. PMID- 17915248 TI - Working harder to stay alive: metabolic rate increases with age in Drosophila simulans but does not correlate with life span. AB - The hypothesis that metabolic rate is inversely correlated with life span has long been debated. Another area of controversy has been the relationship between metabolic rate and aging. In most molecular studies key aspects of cellular metabolism have been shown to decline with age. Less attention has been focused on metabolic rate as an organism ages. We studied the survival of three Drosophila simulans fly lines and measured whole organism metabolic rate, mitochondrial DNA copy number and walking speed. Metabolic rate as assayed by CO(2) production did not correlate with median lifespan but increased by 0.43 1.14%/d. In contrast, mitochondrial DNA copy number decreased by 0.56-1.06%/d. Physical activity, as assayed by mean walking speed, did not change with age but was positively correlated with mitochondrial DNA copy number. One explanation for these data is that metabolic rate was increased, in the face of a reduced mitochondrial DNA copy number and capacity for oxidative metabolism, to maintain a constant bioenergetic demand (physical activity). Alternatively, metabolic rate may increase to provide energy for the repair of cellular damage or due to a shift in metabolic substrate use over time. PMID- 17915249 TI - The Zn2 position in metallo-beta-lactamases is critical for activity: a study on chimeric metal sites on a conserved protein scaffold. AB - Metallo-beta-lactamases (MbetaLs) are bacterial Zn(II)-dependent hydrolases that confer broad-spectrum resistance to beta-lactam antibiotics. These enzymes can be subdivided into three subclasses (B1, B2 and B3) that differ in their metal binding sites and their characteristic tertiary structure. To date there are no clinically useful pan-MbetaL inhibitors available, mainly due to the unawareness of key catalytic features common to all MbetaL brands. Here we have designed, expressed and characterized two double mutants of BcII, a di-Zn(II) B1-MbetaL from Bacillus cereus, namely BcII-R121H/C221D (BcII-HD) and BcII-R121H/C221S (BcII-HS). These mutants display modified environments at the so-called Zn2 site or DCH site, reproducing the metal coordination environments of structurally related metallohydrolases. Through a combination of structural and functional studies, we found that BcII-HD is an impaired beta-lactamase even as a di-Zn(II) enzyme, whereas BcII-HS exhibits the ability to exist as mono or di-Zn(II) species in solution, with different catalytic performances. We show that these effects result from an altered position of Zn2, which is incapable of providing a productive interaction with the substrate beta-lactam ring. These results indicate that the position of Zn2 is essential for a productive substrate binding and hydrolysis. PMID- 17915250 TI - Ligand-induced conformational changes via flexible linkers in the amino-terminal region of the inositol 1,4,5-trisphosphate receptor. AB - Cytoplasmic Ca2+ signals are highly regulated by various ion transporters, including the inositol 1,4,5-trisphosphate (IP(3)) receptor (IP(3)R), which functions as a Ca2+ release channel on the endoplasmic reticulum membrane. Crystal structures of the two N-terminal regulatory regions from type 1 IP(3)R have been reported; those of the IP(3)-binding core (IP(3)R(CORE)) with bound IP(3), and the suppressor domain. This study examines the structural effects of ligand binding on an IP(3)R construct, designated IP(3)R(N), that contains both the IP(3)-binding core and the suppressor domain. Our circular dichroism results reveal that the IP(3)-bound and IP(3)-free states have similar secondary structure content, consistent with preservation of the overall fold within the individual domains. Thermal denaturation data show that, while IP(3) has a large effect on the stability of IP(3)R(CORE), it has little effect on IP(3)R(N), indicating that the suppressor domain is critical to the stability of IP(3)R(N). The NMR data for IP(3)R(N) provide evidence for chemical exchange, which may be due to protein conformational dynamics in both apo and IP(3)-bound states: a conclusion supported by the small-angle X-ray scattering data. Further, the scattering data show that IP(3)R(N) undergoes a change in average conformation in response to IP(3) binding and the presence of Ca2+ in the solution. Taken together, these data lead us to propose that there are two flexible linkers in the N-terminal region of IP(3)R that join stably folded domains and give rise to an equilibrium mixture of conformational sub-states containing compact and more extended structures. IP(3) binding drives the conformational equilibrium toward more compact structures, while the presence of Ca2+ drives it to a more extended set. PMID- 17915251 TI - Structural characterization of a new binding motif and a novel binding mode in group 2 WW domains. AB - Formin homology 1 (FH1), is a long proline-rich region of formins, shown to bind to five WW containing proteins named formin binding proteins (FBPs). FH1 has several potential binding regions but only the PPLPx motif and its interaction with FBP11WW1 has been characterized structurally. To detect whether additional motifs exist in FH1, we synthesized five peptides and investigated their interaction with FBP28WW2, FBP11WW1 and FBP11WW2 domains. Peptides of sequence PTPPPLPP (positive control), PPPLIPPPP and PPLIPPPP (new motifs) interact with the domains with micromolar affinity. We observed that FBP28WW2 and FBP11WW2 behave differently from FBP11WW1 in terms of motif selection and affinity, since they prefer a doubly interrupted proline stretch of sequence PPLIPP. We determined the NMR structure of three complexes involving the FBP28WW2 domain and the three ligands. Depending on the peptide under study, the domain interacts with two proline residues accommodated in either the XP or the XP2 groove. This difference represents a one-turn displacement of the domain along the ligand sequence. To understand what drives this behavior, we performed further structural studies with the FBP11WW1 and a mutant of FBP28WW2 mimicking the XP2 groove of FBP11WW1. Our observations suggest that the nature of the XP2 groove and the balance of flexibility/rigidity around loop 1 of the domain contribute to the selection of the final ligand positioning in fully independent domains. Additionally, we analyzed the binding of a double WW domain region, FBP11WW1-2, to a long stretch of FH1 using fluorescence spectroscopy and NMR titrations. With this we show that the presence of two consecutive WW domains may also influence the selection of the binding mode, particularly if both domains can interact with consecutive motifs in the ligand. Our results represent the first observation of protein-ligand recognition where a pair of WW and two consecutive motifs in a ligand participate simultaneously. PMID- 17915252 TI - The CNTN4 c.4256C>T mutation is rare in Japanese with inherited spinocerebellar ataxia. AB - To confirm the incidence of SCA16 in Japan, we screened DNA samples from a number of patients of ataxia of unknown etiology for the substitution. We examined a total of 323 DNA samples from Japanese patients with inherited spinocerebellar ataxia. We found no 317-base pair band in the patients with ataxia of unknown etiology. It seemed that this mutation (c.4256C>T) is rare in Japanese patients with inherited spinocerebellar ataxia. Mutations in other populations should be analyzed. Pathological examinations and molecular biological examinations are needed to confirm that this mutation is a true cause of SCA16. PMID- 17915253 TI - Intraventricular vascular malformations mimicking tumors: case reports and review of the literature. AB - Vascular malformations of the ventricular system are rare, and their clinical and radiologic characteristics vary depending on the location of the lesions. Many types are described, but a comprehensive summary is lacking. Herein, we add two cases to the literature and review known types of ventricular vascular malformations. One case involved a 37-year-old woman who presented with headache due to hydrocephalus. Computed tomography (CT) and magnetic resonance imaging (MRI) revealed a lesion of the foramen of Monro. The other case involved a 7-year old boy who had dizziness and headache due to hydrocephalus. CT and MRI demonstrated a lesion of the third ventricle. In both cases, the images mimicked those of tumors with or without bleeding. As a result, the malformations were misdiagnosed, though surgical treatment was successful. In both cases, the lesions were proven to be arteriovenous malformations on pathologic evaluation. Correct diagnosis of ventricular vascular malformations is sometimes difficult but essential for good treatment planning. Their incidence is low. However, their bleeding and rebleeding rate is high, and they commonly cause hydrocephalus. Treatment should be timely and based on the type of lesion and its presentation. PMID- 17915254 TI - How useful are anti-neural IgM antibodies in the diagnosis of chronic immune mediated neuropathies? AB - Antibodies against several neural antigens have been associated with different chronic immune-mediated neuropathies but their practical clinical relevance remains unclear. To determine the possible diagnostic usefulness of these antibodies we reviewed the clinical correlate of IgM antibodies to the myelin associated glycoprotein (MAG), sulfatide, the gangliosides GM1, GM2, GD1a and GD1b in 539 consecutive patients examined for neuropathy or related diseases in our Neuropathy Clinics and tested for these antibodies in our laboratory since 1985. 302 patients (56%) had an established diagnosis of definite or possible chronic immune-mediated neuropathy while 237 had a neuropathy of non-immune mediated origin or of unknown aetiology or a closely related disease. Antibodies to one or more antigen were more frequent (chi(2)=63.32; p<0.00001) in patients with chronic immune-mediated neuropathy (37.7%) than with other neuropathy or related diseases (7.2%) and their presence was associated in 87% of the patients with an immune-mediated neuropathy, incrementing by 31% the probability of having this form. Testing for MAG permitted to identify 24.8% of patients with an immune mediated neuropathy, GM1 an additional 9.9%, while GM2, GD1b, GD1a and sulfatide altogether an additional 3% of the patients. Concerning clinical correlations, all 75 patients with anti-MAG IgM had neuropathy and IgM monoclonal gammopathy (PN+IgM) with a positive predictive value for this neuropathy of 100%. A similarly high predictive value for neuropathy (91.4%) was observed among 269 patients with IgM monoclonal gammopathy including 103 patients without neuropathy. Anti-sulfatide IgM, though rare, were also significantly and constantly associated with PN+IgM and permitted to identify few patients not bearing anti-MAG IgM. Anti-GM1 IgM were significantly associated with multifocal motor neuropathy (MMN) (29.2%) but where also found in a few patients with other immune or non-immune neuropathies or related diseases with a positive predictive value for MMN of 25.5%. Anti-GM2 IgM were also significantly associated with MMN and increased the sensitivity (36.2%) for MMN obtained with anti-GM1 IgM only, without affecting its specificity and positive predictive value. Anti-GD1a, GD1b, though not significantly more frequent in patients with immune-mediated neuropathy, were associated in 80 to 100% of patients with these neuropathies. In conclusion anti-neural IgM antibodies may help in identifying patients with a chronic immune-mediated neuropathy, even if only anti-MAG and anti-sulfatide IgM appear to be strictly associated with a definite clinical syndrome. PMID- 17915255 TI - Oxidative stress is induced in female carriers of X-linked adrenoleukodystrophy. AB - X-linked adrenoleukodystrophy (X-ALD) is a peroxisomal disease biochemically characterized by the accumulation of very long chain fatty acids (VLCFA), particularly hexacosanoic (C26:0) and tetracosanoic acids (C24:0) in different tissues and in biological fluids and clinically characterized by central and peripheral demyelination and adrenal insufficiency. A considerable number of heterozygotes (HTZ) for X-ALD develop neurological symptoms like spinal cord involvement resembling milder forms of adrenomyeloneuropathy. However, the mechanisms of brain damage in hemizygotes and heterozygotes X-ALD individuals are poorly understood. Considering that oxidative stress was involved in various neurodegenerative disorders and that in a previous study we showed evidence that oxidative stress is probably involved in the pathophysiology of X-ALD symptomatic patients, in the present study we evaluated various oxidative stress parameters, namely thiobarbituric acid-reactive substances (TBA-RS), total antioxidant status (TAS) and total antioxidant reactivity (TAR) in plasma of HTZ individuals for X ALD. It was observed that female carriers present a significant increase of TBA RS measurement, indicating a stimulation of lipid peroxidation, as well as a decrease of TAR, reflecting a deficient capacity to rapidly handle an increase of reactive species. These results indicate that oxidative stress is involved in the pathophysiology of heterozygotes for X-ALD. PMID- 17915256 TI - Gene-based modelling for rice: an opportunity to enhance the simulation of rice growth and development? AB - Process-based crop simulation models require employment of new knowledge for continuous improvement. To simulate growth and development of different genotypes of a given crop, most models use empirical relationships or parameters defined as genetic coefficients to represent the various cultivar characteristics. Such a loose introduction of different cultivar characteristics can result in bias within a simulation, which could potentially integrate to a high simulation error at the end of the growing season when final yield at maturity is predicted. Recent advances in genetics and biomolecular analysis provide important opportunities for incorporating genetic information into process-based models to improve the accuracy of the simulation of growth and development and ultimately the final yield. This improvement is especially important for complex applications of models. For instance, the effect of the climate change on the crop growth processes in the context of natural climatic and soil variability and a large range of crop management options (e.g., N management) make it difficult to predict the potential impact of the climate change on the crop production. Quantification of the interaction of the environmental variables with the management factors requires fine tuning of the crop models to consider differences among different genotypes. In this paper we present this concept by reviewing the available knowledge of major genes and quantitative trait loci (QTLs) for important traits of rice for improvement of rice growth modelling and further requirements. It is our aim to review the assumption of the adequacy of the available knowledge of rice genes and QTL information to be introduced into the models. Although the rice genome sequence has been completed, the development of gene-based rice models still requires additional information than is currently unavailable. We conclude that a multidiscipline research project would be able to introduce this concept for practical applications. PMID- 17915257 TI - Microarrays and the relationship of mRNA variation to protein variation during the cell cycle. AB - Microarray analyses have led to the postulated existence and identification of numerous genes that are believed to be expressed and presumably to act in a cell cycle-specific manner because their expression varies during the cell cycle. It is important to see how protein variation can be produced from mRNA variation. We have calculated the protein content throughout the cell cycle resulting from cell cycle-specific mRNA expression, and compared the result to protein content resulting from constant, cell-cycle independent, mRNA expression. For stable proteins, cell-cycle-specific mRNA expression leads to a maximum 2-fold change in protein content compared to proteins synthesized from constantly expressed mRNA. More realistic sinusoidal patterns of mRNA expression exhibit much smaller ratios of 1.25 or lower, even for extremely large amplitudes in mRNA expression. For unstable proteins that have a cycle-independent half-life, only at extremely short protein half-lives does mRNA variation have a significant impact on variation of protein content during the division cycle. We also apply these findings to proteins with a cycle-specific decay pattern. mRNA variations during the eukaryotic division cycle variation of mRNA during the cell cycle can have only a minimal affect on the variation of protein content during the cell cycle. We conclude that mRNA variations during the division cycle, as measured by microarrays, cannot by themselves, identify cycle-specific functions related to protein variations. PMID- 17915258 TI - Reactive oxygen species trigger ischemic and pharmacological postconditioning: in vivo and in vitro characterization. AB - Reactive oxygen species (ROS) generated by ischemic and pharmacological preconditioning are known to act as triggers of cardiac protection; however, the involvement of ROS in ischemic and pharmacological postconditioning (PostC) in vivo and in vitro is unknown. We tested the hypothesis that ROS are involved in PostC in the mouse heart in vivo and in the isolated adult cardiac myocyte (ACM). Mice were subjected to 30 min coronary artery occlusion followed by 2 h of reperfusion with or without ischemic or pharmacologic PostC (three cycles of 20 s reperfusion/ischemia; 1.4% isoflurane; 10 mg/kg SNC-121). Additional groups were treated with 2-mercaptopropionyl glycine (MPG), a ROS scavenger, 10 min before or after the PostC stimuli. Ischemia-, isoflurane-, and SNC-121- induced PostC reduced infarct size (24.1+/-3.2, 15.7+/-2.6, 24.9+/-2.6%, p<0.05, respectively) compared to the control group (43.4+/-3.3%). These cardiac protective effects were abolished by MPG when administered before (40.0+/-3.6, 39.3+/-3.1, 38.5+/ 1.6%, respectively), but not after the PostC stimuli (26.6+/-2.3, 17.0+/-2.2, 23.9+/-1.7%, respectively). Additionally, ACM were subjected to a simulated ischemia/reperfusion protocol with isoflurane and SNC PostC. Isoflurane- and SNC induced PostC in vitro were abolished by prior treatment with MPG. These data indicate that ROS signaling is an essential trigger of ischemic and pharmacological PostC and this is occurring at the level of the cardiac myocyte. PMID- 17915259 TI - Inhibitory effect of naringenin chalcone on inflammatory changes in the interaction between adipocytes and macrophages. AB - Obese adipose tissue is characterized by an enhanced infiltration of macrophages. It is considered that the paracrine loop involving monocyte chemoattractant protein (MCP)-1 and tumor necrosis factor (TNF)-alpha between adipocytes and macrophages establishes a vicious cycle that augments the inflammatory changes and insulin resistance in obese adipose tissue. Polyphenols, which are widely distributed in fruit and vegetables, can act as antioxidants and some of them are also reported to have anti-inflammatory properties. Tomato is one of the most popular and extensively consumed vegetable crops worldwide, which also contains many flavonoids, mainly naringenin chalcone. We investigated the effect of flavonoids, including naringenin chalcone, on the production of proinflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophages and in the interaction between adipocytes and macrophages. Naringenin chalcone inhibited the production of TNF-alpha, MCP-1, and nitric oxide (NO) by LPS-stimulated RAW 264 macrophages in a dose-dependent manner. Coculture of 3T3-L1 adipocytes and RAW 264 macrophages markedly enhanced the production of TNF-alpha, MCP-1, and NO compared with the control cultures; however, treatment with naringenin chalcone dose-dependently inhibited the production of these proinflammatory mediators. These results indicate that naringenin chalcone exhibits anti-inflammatory properties by inhibiting the production of proinflammatory cytokines in the interaction between adipocytes and macrophages. Naringenin chalcone may be useful for ameliorating the inflammatory changes in obese adipose tissue. PMID- 17915261 TI - A novel mutation of p63 in a Chinese family with inherited syndactyly and adactylism. AB - p63 is a transcription factor homologous to p53 and p73; mutations in this gene have been identified in individuals with several types of developmental abnormalities, including EEC (ectrodactyly, ectodermal dysplasia, facial clefts) syndrome and split-hand/split-foot malformation (SHFM). Several mutations in the p63 gene have previously been shown to be related to SHFM. In this study, we report on a Chinese family with intrafamilial clinical variability of SHFM that have a novel heterozygous mutation in all four affected individuals. The mutation is in exon 8 of p63, 1046G --> A, which predicts an amino acid substitution G310E. SSCP analysis of the segregation pattern of the mutation strongly suggests a causal relationship to the SHFM phenotype in p63. This mutation has not been observed in other countries in the world. PMID- 17915260 TI - Nitric oxide signaling participates in norepinephrine-induced activity of neuronal intracellular survival pathways. AB - Much evidence has gathered that nitric oxide (NO) signaling, via cGMP-dependent mechanisms, may activate pro-survival pathways in hippocampal neurons and inhibit apoptosis. Past research has revealed that the enhancement of monoaminergic neurotransmission via exercise or treatment with antidepressant medications leads to an enhanced expression of brain-derived neurotrophic factor (BDNF). In isolated hippocampal neurons, norepinephrine (NE) application also increases the immunoreactivity of BDNF and several pro-survival signaling molecules. The data herein support the possibility that NO signaling plays an important role in enhancing neurotrophin expression and activation of the pro-survival phosphatidylinositol 3' kinase (PI-3K) pathway stimulated by NE. In isolated hippocampal neurons, the NO donor, sodium nitroprusside, increases BDNF, PI-3K, and phospho-ERK1 immunoreactivity. Specific inhibitors of the NO system suggest that NE-induced increases in hippocampal BDNF and the PI-3K pathway, but not stimulation of the MAPK pathway, depend upon NO signaling. In addition, inhibiting cGMP suggest that the effects of NE on BDNF immunoreactivity and Akt phosphorylation are also cGMP-dependent. Finally, the application of l-NAME to hippocampal neurons increases cell death. This is the first study of its kind demonstrating the involvement of NE-induced pro-survival signaling in three distinct signaling pathways: PI-3K, MAPK, and NO/cGMP. Possible mechanisms are discussed in light of the results. PMID- 17915262 TI - Frontal eye fields involved in shifting frame of reference within working memory for scenes. AB - Working memory (WM) evoked by linguistic cues for allocentric spatial and egocentric spatial aspects of a visual scene was investigated by correlating fMRI BOLD signal (or "activation") with performance on a spatial-relations task. Subjects indicated the relative positions of a person or object (referenced by the personal pronouns "he/she/it") in a previously shown image relative to either themselves (egocentric reference frame) or shifted to a reference frame anchored in another person or object in the image (allocentric reference frame), e.g. "Was he in front of you/her?" Good performers had both shorter response time and more correct responses than poor performers in both tasks. These behavioural variables were entered into a principal component analysis. The first component reflected generalised performance level. We found that the frontal eye fields (FEF), bilaterally, had a higher BOLD response during recall involving allocentric compared to egocentric spatial reference frames, and that this difference was larger in good performers than in poor performers as measured by the first behavioural principal component. The frontal eye fields may be used when subjects move their internal gaze during shifting reference frames in representational space. Analysis of actual eye movements in three subjects revealed no difference between egocentric and allocentric recall tasks where visual stimuli were also absent. Thus, the FEF machinery for directing eye movements may also be involved in changing reference frames within WM. PMID- 17915263 TI - Longitudinal aspects of emotion recognition in patients with traumatic brain injury. AB - Changes in emotional and social behaviour are relatively common following traumatic brain injury (TBI). Impairments in recognising the emotional state of others may underlie some of the problems in social relationships that these patients experience. The few previous studies examining emotion recognition in TBI typically assessed patients once, long after the onset of brain injury, making it difficult to distinguish the direct effect of brain injury from the effects of environmental changes. This study examined 30 patients with TBI shortly after brain injury and 32 orthopaedic control patients on their recognition of emotions expressed in the face and the voice using discrimination and labelling tasks. These patients were followed up 1 year later to examine the longitudinal development of emotion recognition deficits. TBI patients were found to be impaired on emotion recognition compared to the control patients both early after injury and 1 year later. The fact that impairments in emotion recognition were evident early after TBI and no evidence of recovery over time was found, suggests a direct effect of brain injury. PMID- 17915264 TI - Neural correlates of emotional working memory in patients with mild cognitive impairment. AB - Emotional stimuli can have beneficial effects on memory in healthy aged subjects and partly on patients with dementia. So far, no experimental study has explored the effects of memory for emotional stimuli in patients with mild cognitive impairment (MCI), a concept that describes a transitional state between normal aging and dementia. The present fMRI study explored working memory for emotional stimuli in 16 patients with amnestic MCI (aMCI) and 16 healthy aged participants. Subjects performed an n-back task (2-back) with neutral, positive, and negative emotional pictures. The analysis focused on target processing. Results showed that groups did not differ in working memory performance. In healthy aged participants emotional targets had no significant impact on working memory. In patients with aMCI a negativity bias was observed, indicating that negative targets were better remembered compared to neutral and positive targets. Regarding fMRI results, both groups showed an increase in functional activity in prefrontal and lateral parietal brain regions associated with target processing. As a key result, we observed significant group by emotion interaction effects in the precuneus. Healthy aged participants showed a signal decrease in the left precuneus for positive compared to neutral targets. The precuneus deactivation in healthy aged participants may indicate a disengagement of self-referential processes towards task-related processes. Patients with aMCI revealed a signal increase in the right precuneus for negative compared to neutral targets. This increase in precuneus activity, combined with a behavioural facilitation effect, may indicate a mechanism to compensate disease related processes in aMCI. PMID- 17915266 TI - Awakening cortisol response in relation to psychosocial profiles and eating behaviors. AB - Awakening cortisol response was measured in 78 men and women, on 3 mornings within a 2-month period. Psychosocial and eating behavior variables were assessed using self-administered questionnaires on anxiety (State-Trait Anxiety Inventory), depression (Beck Depression Inventory), body esteem (Body Esteem Scale for Adolescents and Adults), and eating behaviors (Three-Factor Eating Questionnaire and Eating Disorder Inventory-2). Data on food intake and appetite sensations were also collected using a buffet-type meal test, a 3-day food record and visual analog scales measured before and after a standardized breakfast meal test. In women, high anxiety, disinhibition and hunger scores, as well as poor body esteem and a high weight preoccupation, were negatively correlated to ACR. The factor that appeared to account the most for this inverse relation was emotional susceptibility to disinhibition (r=-0.61, p=0.003). The latter was also negatively associated with the satiety quotient for fullness in response to the standardized breakfast (r=-0.48, p=0.010). In men, ACR was negatively associated with flexible (r=-0.33, p=0.020) and strategic (r=-0.28, p=0.049) restraint behaviors. This study highlights a gender-dependent relationship between ACR, hence the activity of the hypothalamic-pituitary-adrenal axis, and eating behaviors and psychological profiles. PMID- 17915267 TI - Malaria drug and vaccine trials in Africa: obstacles and opportunities. AB - There are several new treatments and vaccine technologies in clinical development for childhood malaria that have arrived in the clinical phase of evaluation during the past 5-10 years. This is a long-awaited change as until this time there had been little in the pipeline. As these products progress, evaluating them in the populations for whom they are being developed is becoming increasingly challenging. Many more capable trial sites are required and thousands of children and their parents need to be willing to take part in all the clinical trials that will be necessary if even a handful of these products make it through to obtaining a marketing approval license. Then, beyond licensure, these products will need to be assessed in more 'real-life' phase IV trials to establish whether they can truly impact the high level of mortality that malaria brings to the under-five population in Africa. Here we explore the issues that face both the trial sites and the product developers and present how this opportunity should be utilised to develop experienced African clinical researchers and facilities alongside getting these products through into public health use. PMID- 17915265 TI - Deafness for the meanings of number words. AB - We describe the performance of an aphasic individual who showed a selective impairment affecting his comprehension of auditorily presented number words and not other word categories. His difficulty in number word comprehension was restricted to the auditory modality, given that with visual stimuli (written words, Arabic numerals and pictures) his comprehension of number and non-number words was intact. While there have been previous reports of selective difficulty or sparing of number words at the semantic and post-semantic levels, this is the first reported case of a pre-semantic deficit that is specific to the category of number words. This constitutes evidence that lexical semantic distinctions are respected by modality-specific neural mechanisms responsible for providing access to the meanings of words. PMID- 17915268 TI - Risk factors associated with St. Louis encephalitis seroprevalence in two populations from Cordoba, Argentina. AB - St. Louis encephalitis virus (SLEV) is endemic in Argentina. The first outbreak was recorded in Cordoba during the late summer of 2005. This study analyzes the association between social and demographic factors and other variables related to exposure to mosquitoes with SLEV infection in 264 individuals who attended two health centers in the city of Cordoba during the period December 2004-January 2005. Multiple logistic regression models were used to estimate the risk coefficients. The seroprevalence was 12.9%, similar to that in previous reports. Risks of infection were associated with the presence of garbage dumps near dwellings, the practice of outdoor activities at night and place of residence. Risk for older people (60-80 years old) was moderate. The identification of risk factors related to SLEV infection would be useful to improve programs for vector control and community health. PMID- 17915269 TI - Modelling sexually transmitted infections: less is usually more for informing public health policy. AB - Mathematical models have been used to investigate the dynamics of infectious disease transmission since Bernoulli's smallpox modelling in 1760. Their use has become widespread for exploring how epidemics can be prevented or contained. Here we discuss the importance of modelling the dynamics of sexually transmitted infections, the technology-driven dichotomy in methodology, and the need to 'keep it simple' to explore sensitivity, to link the models to reality and to provide understandable mechanistic explanations for real-world policy-makers. The aim of models, after all, is to influence or change public health policy by providing rational forecasting based on sound scientific principles. PMID- 17915270 TI - Pilot study of sodium phenylbutyrate as adjuvant in cyclophosphamide-resistant endemic Burkitt's lymphoma. AB - Burkitt's lymphoma (BL) accounts for the majority of childhood malignancies seen in sub-Saharan Africa. In Malawi, cyclophosphamide (CPM), the mainstay of treatment for endemic BL, is effective in around 50% of cases. Evidence exists in support of an association between activation of replication of Epstein-Barr virus (EBV) in the tumour and response to this chemotheraupeutic agent. Phenylbutyrate (PB), approved for treatment of inborn errors of the urea cycle with minimal toxicity in children, induces EBV replication and cell lysis in BL-derived cell cultures. It has also shown some success as adjuvant in treatment of chronic leukaemia and lymphoma. We tested in African BL patients with CPM-resistant tumours, and thus unlikely to survive, the hypothesis that PB can reverse this resistance. A study of five patients showed PB before CPM to induce shrinkage of CPM-resistant tumours in two of them. Findings suggested that for this effect PB pre-treatment should be given for a week before CPM treatment. A larger study is indicated. PMID- 17915271 TI - Helpful diagnostic markers of steroidogenesis for defining hyperandrogenemia in hirsute women. AB - HYPOTHESIS: Androgen excess carries varied clinical manifestations in women. Although testosterone and dehydroepiandrostendionesulfate (DHEAS) determination is considered useful in diagnostic workup, there is no laboratory definition that sufficiently describes androgen excess. DESIGN: We studied 464 hirsute women with a Ferriman and Gallwey score of at least 8 between 2000 and 2005. Our examination included clinical data, total testosterone (T), sex hormone-binding globulin (SHBG), the free androgen index (FAI), and DHEAS. Additionally, androstendione, 17alpha-hydroxyprogesterone (17OHP), dehydroepiandrostendione (DHEA), and 11 deoxycortisol were determined at baseline and 60min after corticotropin challenge (250microg synacthen). RESULTS: Of 464 women, 77.6% fulfilled the clinical criteria for hyperandrogenemia. Of these 360 women, 78.1% had hyperandrogenic hirsutism. Of these 281 women, 43.4% showed increased stimulation of 17OHP to 250microg of synacthen. Another 37.4% showed adrenal steroid biosynthesis defects other than 21alpha-hydroxylase deficiency, such as defective 11beta-hydroxylation or 3beta-hydroxysteroid dehydrogenase malfunction. The diagnosis of polycystic ovary syndrome was applicable to 12.4%. In addition, our results show that 72% of 281 patients with secondary hirsutism had normal T concentrations, and 55% had a normal FAI. Only 5% of hirsute patients with a normal FAI had elevated DHEAS values. However, 40% showed elevated DHEA levels, while 26% of the women with normal FAI showed androstendione values over the maximal levels in the 79 controls. CONCLUSIONS: Our data suggest that in addition to testosterone and FAI, androstendione and DHEA are significantly helpful parameters in diagnosing hyperandrogenemia in hirsute women. DHEAS was not found to be helpful. PMID- 17915272 TI - Coexistence of cytoplasmic incompatibility and male-killing-inducing endosymbionts, and their impact on host gene flow. AB - Male-killing (MK) and cytoplasmic incompatibility (CI) inducing bacteria are among the most common endosymbionts of arthropods. Previous theoretical research has demonstrated that these two types of endosymbionts cannot stably coexist within a single unstructured host population if no doubly infected host individuals occur. Here, we analyse a model of two host subpopulations connected by migration. We demonstrate that coexistence of MK- and CI-inducing endosymbionts is possible if migration rates are sufficiently low. In particular, our results suggest that for coexistence to be possible, migration rates into the subpopulation infected predominantly with MK-inducing endosymbionts must be considerably low, while migration rates from the MK- to the CI-infected subpopulation can be very high. We also analyse how the presence of MK- and CI inducing endosymbionts affects host gene flow between the two subpopulations. Employing the concept of the 'effective migration rate', we demonstrate that compared with an uninfected subdivided population, gene flow is increased towards the MK-infected island, but decreased towards the CI-infected island. We discuss our results with respect to the butterfly Hypolimnas bolina, in which infection polymorphism of CI- and MK-inducing Wolbachia has been reported across South Pacific island populations. PMID- 17915274 TI - Stochastic modeling of cellular colonies with quiescence: an application to drug resistance in cancer. AB - Several cancers are thought to be driven by cells with stem cell like properties. An important characteristic of stem cells, which also applies to primitive tumor cells, is the ability to undergo quiescence, where cells can temporarily stop the cell cycle. Cellular quiescence can affect the kinetics of tumor growth, and the susceptibility of the cells to therapy. To study how quiescence affects treatment, we formulate a stochastic birth-death process with quiescence, on a combinatorial cellular mutation network, and consider the pre-treatment (growth) and treatment (decay) regimes. We find that, in the absence of mutations, treatment (if sufficiently strong) will proceed as a biphasic decline with the first (faster) phase driven by the elimination of the cycling cells and the second (slower) phase limited by the process of cell awakening. Other regimes are possible for weaker treatments. We also describe how the process of mutant generation is influenced by quiescence. Interestingly, for single-drug treatments, the probability to have resistance at start of treatment is independent of quiescence. For two or more drugs, the probability to have generated resistant mutants before treatment grows with quiescence. Finally, we study the influence of quiescence on the treatment phase. Starting from a given composition of mutants, the chances of treatment success are not influenced by the presence of quiescence. PMID- 17915275 TI - Immunohistochemical detection of matrix metalloproteinase-1 in the periodontal ligament of equine cheek teeth. AB - The hypsodont equine cheek tooth erupts continuously throughout life. The collagen fibers of the periodontal ligament (PDL) have to remodel constantly to allow the tooth to move in an occlusal direction. Remodeling of the collagen fiber bundles needs to be well-coordinated in order to maintain functional tooth support. The aim of this study was to examine the role of matrix metalloproteinase-1 (MMP-1) in the collagen remodeling of the equine PDL under physiological conditions. Specimens containing the PDL interposed between the dental cementum and the alveolar bone were taken from nine Warmblood horses at three designated horizontal levels: subgingival, middle, and apical. The expression of MMP-1 was detected immunohistochemically. MMP-1 was found to be present in the specimens of all horses. Immunopositive fibroblasts/fibrocytes were accumulated within individual single collagen fascicles. Our results suggest that MMP-1 induced collagen degradation plays a central role in the physiological remodeling of the equine PDL. The distribution of MMP-1 positive fascicles indicates well-directed remodeling which occurs as an asynchronous process, so that only single collagen fascicles are remodeled at the same time. Due to this remodeling of one fascicle at a time, the overall anchorage of the tooth is preserved at all times. PMID- 17915273 TI - The evolution of bet-hedging adaptations to rare scenarios. AB - When faced with a variable environment, organisms may switch between different strategies according to some probabilistic rule. In an infinite population, evolution is expected to favor the rule that maximizes geometric mean fitness. If some environments are encountered only rarely, selection may not be strong enough for optimal switching probabilities to evolve. Here we calculate the evolution of switching probabilities in a finite population by analyzing fixation probabilities of alleles specifying switching rules. We calculate the conditions required for the evolution of phenotypic switching as a form of bet-hedging as a function of the population size N, the rate theta at which a rare environment is encountered, and the selective advantage s associated with switching in the rare environment. We consider a simplified model in which environmental switching and phenotypic switching are one-way processes, and mutation is symmetric and rare with respect to the timescale of fixation events. In this case, the approximate requirements for bet-hedging to be favored by a ratio of at least R are that sN>log(R) and thetaN>square root R . PMID- 17915276 TI - Inhibitory effect of crotoxin on the pain-evoked discharge of neurons in thalamic parafascicular nucleus in rats. AB - Crotoxin (Cro), the principal neurotoxic component of Crotalus durissus terrificus, has been previously reported to have a behavioral analgesic effect in rats and mice. The present study investigated electrophysiologically the effect of Cro on pain-evoked unit discharge of neurons in thalamic parafascicular nucleus (Pf) and underlying mechanisms of its effect. The electrical discharge of Pf neurons was recorded with the microelectrode technique in rats. Intracerebroventricular (i.c.v.) injection of Cro at 0.25, 0.45 and 0.65 microg/kg resulted in a dose-dependent inhibitory effect on the pain-evoked discharge of Pf neurons. The discharge frequency and the discharge duration significantly (P<0.05) decreased after Cro administration. This inhibitory effect was significantly (P<0.05) attenuated after pretreatment with para chlorophenylalanine (pCPA), or electrolytic lesion of dorsal raphe (DR) nucleus. In contrast, i.c.v. injection of atropine (muscarinic receptor antagonist, 5 microg) or naloxone (opioid receptor antagonist, 4 microg) had no effect on Cro induced inhibition of discharge of Pf neurons. The results suggested that Cro has an analgesic effect, which is mediated, at least partially, by the central serotonergic system. PMID- 17915277 TI - Systemic and local myotoxicity induced by snake venom group II phospholipases A2: comparison between crotoxin, crotoxin B and a Lys49 PLA2 homologue. AB - The patterns of myotoxicity induced in mice by crotoxin, crotoxin B and a Lys49 phospholipase A(2) (PLA(2)) homologue were compared. Lys49 PLA(2)-induced local myotoxicity is reflected by creatine kinase (CK) loss in injected gastrocnemius muscle, and by a profile of CK increase in plasma characterized by a rapid increment and drop after intramuscular injection, and by a lack of CK increase in plasma after intravenous injection. In contrast, crotoxin and crotoxin B, which induce local and systemic myotoxicity, provoked a more prolonged increment in plasma CK activity upon intramuscular injection, and induced increments in plasma CK after intravenous injection. The three toxins promoted a similar extent of local myotoxicity, assessed by the loss of CK in injected gastrocnemius. A method for the quantitative assessment of the ability of toxins to induce systemic myotoxicity is proposed, based on the estimation of the ratio between the area under the curve in the plasma CK activity (total myotoxicity) to the loss of CK in injected gastrocnemius (local myotoxicity). The highest ratio corresponded to crotoxin, and the lowest corresponded to Lys49 PLA(2), the former being a systemic myotoxin and the latter a local myotoxin. Neutralization by antivenoms also differed between the toxins: a drastic reduction in plasma CK, with very poor neutralization of local CK loss, was achieved in the case of crotoxin B when antivenom was injected intravenously, whereas no neutralization was achieved in the case of Lys49 PLA(2). When tested in undifferentiated myoblasts in culture, Lys49 PLA(2) induced cytotoxicity, whereas crotoxin and crotoxin B did not, evidencing that the latter are devoid of widespread cytolytic activity. Molecular modeling analysis showed that Lys49 PLA(2) has a conspicuous cationic face, which is likely to interact with diverse membranes. In contrast, crotoxin B, despite its overall basic pI, has a lower density of positively charged residues at this molecular region. It is suggested that Lys49 PLA(2)s homologues interact, through this cationic face, with many different cell types, thus lacking specificity for muscle cells. In contrast, crotoxin B has a more selective interaction with targets in the muscle cell membrane. This selectivity might be the basis for the ability of crotoxin and crotoxin B to induce systemic myotoxicity. PMID- 17915278 TI - Immunization with a replication-defective herpes simplex virus 2 mutant reduces herpes simplex virus 1 infection and prevents ocular disease. AB - Ocular infections with herpes simplex virus 1 can lead to corneal scarring and blindness, with herpes keratitis being the major infectious cause of blindness. There is currently no clinically approved vaccine and nearly all developmental vaccines are targeted against HSV-2 and genital herpes. We tested the ability of an HSV-2 replication-defective virus, a genital herpes vaccine candidate, to protect against HSV-1 corneal infection. Immunization with HSV-2 dl5-29 reduced viral replication in the cornea, prevented ocular disease and reduced latent infection by the HSV-1 strain. Therefore, this HSV-2 replication-defective mutant strain may have applications for prevention of herpes keratitis and genital herpes due to HSV-1 infection. PMID- 17915279 TI - Equilibrium dialysis and ultrafiltration investigations of perchlorate removal from aqueous solution using poly(diallyldimethylammonium) chloride. AB - Use of perchlorate salts in military activities and the aerospace industry is widespread. These salts are highly water-soluble and are, to a large extent, kinetically inert as aqueous species. As a groundwater contaminant, perchlorate is now being detected in an increasing number of locations and is believed to interfere with the uptake of iodide by the thyroid, which can result in decreased hormone production. The United States Environmental Protection Agency (US EPA) has established a reference dose for perchlorate of 0.0007 mg/kg/day, which translates to a drinking water equivalent level of 24.5 ppb. This study investigated the application of polyelectrolyte-enhanced ultrafiltration (PEUF) for the selective removal of perchlorate from aqueous solution through equilibrium dialysis and ultrafiltration experiments. Using poly(diallyldimethylammonium) chloride, the effectiveness and efficiency of PEUF in the removal of perchlorate from other aqueous solution components was investigated by testing parameters such as polyelectrolyte concentration, pH, and ionic strength. Removal of perchlorate from synthetic groundwater initially containing 10.3 ppm perchlorate and also containing chloride, sulfate, and carbonate was also examined. Perchlorate separations of greater than 95% were achieved, even in the presence of 10-fold excesses of competing ions. PMID- 17915280 TI - Heterotrophic activity compromises autotrophic nitrogen removal in membrane aerated biofilms: results of a modeling study. AB - A 1-d multi-population biofilm model was constructed to study the effect of heterotrophic activity on completely autotrophic ammonium (NH4+) removal in membrane-aerated (counter-diffusion) versus conventional biofilm systems (co diffusion). Growth of heterotrophic bacteria (HB) was supported either solely by biomass decay products or by organic carbon (as chemical oxygen demand (COD)) in the influent. Three scenarios were considered: influence of HB growing on biomass decay products on steady-state performance (total nitrogen (TN) removal efficiency); influence of the influent COD/N ratio on steady-state performance (supplying COD in the influent); and impact of dynamic changes in the influent COD/N ratio on TN removal efficiency. The results revealed that the TN removal efficiency in the counter-diffusion biofilm was significantly different when HB were included in the simulations at NH4+ surface loads of LNH4>2.7 g - N m(-2) d( 1). Influent COD significantly altered the microbial community composition in the counter-diffusion biofilm and anaerobic NH4+ oxidation could not be sustained at COD/N>2. The co-diffusion system, however, was less affected and more than 50% of the TN removal originated from anaerobic NH4+ oxidation at those ratios. Perturbation experiments showed that step increases to influent COD/N ratios of 2 or higher over a period of 50 d or longer caused a loss of anaerobic NH4+ oxidation capacity which could not be regained within a reasonable time frame (>1000 d) in the counter-diffusion system. In contrast, simulating a 1-d sloughing event only caused a disturbance of 200 d although a maximum biofilm loss of 90-95% occurred. These results clearly indicate the importance of heterotrophic activity in autotrophic N removal biofilms, especially in counter diffusion systems where they may compromise N removal capacity. PMID- 17915281 TI - A clay-vesicle system for water purification from organic pollutants. AB - Vesicle-clay complexes in which positively charged vesicles composed of didodecyldimethylammonium bromide (DDAB) were adsorbed on montmorillonite removed efficiently anionic (sulfentrazone, imazaquin) and neutral (alachlor, atrazine) pollutants from water. These complexes (0.5% w:w) removed 92-100% of sulfentrazone, imazaquin and alachlor and 60% of atrazine from a solution containing 10mg/L of it. A synergistic effect on the adsorption of atrazine was observed when all pollutants were present simultaneously (30 mg/L each), its percentage of removal being 85.5. Column filters (18 cm) filled with a mixture of quartz sand and vesicle-clay (100:1, w:w) were tested. For the passage of 1L (25 pore volumes) of a solution including all the pollutants at 10mg/L each, removal was complete for sulfentrazone and imazaquin, 94% for alachlor and 53.1% for atrazine, whereas removal was significantly less efficient when using activated carbon. A similar advantage of the vesicle-clay filter was observed for the capacities of removal. PMID- 17915282 TI - Microbial ecology of a perchlorate-reducing, hydrogen-based membrane biofilm reactor. AB - The hydrogen-based membrane biofilm reactor (MBfR) has been shown to reduce perchlorate to below 4 microg/L, but little is known about the microbial ecology of this or other hydrogen-based reactors, especially when influent perchlorate concentrations are much lower than the influent oxygen and nitrate concentrations. Dissimilatory (per)chlorate-reducing bacteria (PCRB) can use oxygen as an electron acceptor, and most can also use nitrate. Since oxygen and nitrate can be reduced concurrently with perchlorate, they may serve as primary electron acceptors, sustaining PCRB when the perchlorate concentrations are very low. We studied five identical MBfRs, all seeded with the same inoculum and initially supplied with oxygen, or oxygen plus nitrate, in the influent. After 20 days, perchlorate was added to four MBfRs at influent concentrations of 100 10,000 microg/L, while the fifth was maintained as a control. One day after perchlorate addition, the MBfRs displayed limited perchlorate reduction, suggesting a low initial abundance of PCRB. However, perchlorate reduction improved significantly over time, and denaturing gradient gel electrophoresis (DGGE) analyses suggested an increasing abundance of a single Dechloromonas species. Fluorescence in-situ hybridization (FISH) tests showed that the Dechloromonas species accounted for 14% of the bacterial count in the control MBfR, and 22%, 31%, and 49% in the MBfRs receiving nitrate plus 100, 1000, and 10,000 microg/L perchlorate, respectively. The abundance was 34% in the MBfR receiving oxygen plus 1000 microg/L perchlorate. These results suggest that oxygen is more favorable than nitrate as a primary electron acceptor for PCRB, that PCRB are present at low levels even without perchlorate, and that the presence of perchlorate, even at low levels relative to nitrate or oxygen, significantly enhances selection for PCRB. PMID- 17915283 TI - Variation among plant species in pollutant removal from stormwater in biofiltration systems. AB - Biofiltration systems use vegetation to improve efficiency of pollutant removal from stormwater, but little is known of how plants vary in their capacity to improve biofilter effectiveness. We used a pot trial of 20 Australian species to investigate how species vary in the removal of pollutants from semisynthetic storm water passing through a soil filter medium. Effluent levels of total suspended solids (TSS), Al, Cr, Cu, Pb and Zn were similarly low for vegetated and non-vegetated soils, with reduction to <1-12% of levels in the stormwater input. N and P effluent concentrations were generally lower from vegetated than non-vegetated soils, but total N increased on average in effluent of both vegetated and non-vegetated soils relative to stormwater input. Effluent concentrations varied 2-4-fold among species for TSS, total N and P, total dissolved N (TDN), organic nitrogen and Cu, to more than 20-fold for NOx, NH4+, Mn, Pb and Fe. Species also varied markedly in pollutant removal per root mass (a means of standardising for plant size), with 18-50-fold variation among species in effluent concentrations of total P and N, TDN and organic N, to >150-fold variation in NOx and NH4+. Hence, choice of plant species may have marked effects on biofilter effectiveness. PMID- 17915284 TI - Reactions of chlorine with inorganic and organic compounds during water treatment Kinetics and mechanisms: a critical review. AB - Numerous inorganic and organic micropollutants can undergo reactions with chlorine. However, for certain compounds, the expected chlorine reactivity is low and only small modifications in the parent compound's structure are expected under typical water treatment conditions. To better understand/predict chlorine reactions with micropollutants, the kinetic and mechanistic information on chlorine reactivity available in literature was critically reviewed. For most micropollutants, HOCl is the major reactive chlorine species during chlorination processes. In the case of inorganic compounds, a fast reaction of ammonia, halides (Br(-) and I(-)), SO(3)(2-), CN(-), NO(2)(-), As(III) and Fe(II) with HOCl is reported (10(3)-10(9)M(-1)s(-1)) whereas low chlorine reaction rates with Mn(II) were shown in homogeneous systems. Chlorine reactivity usually results from an initial electrophilic attack of HOCl on inorganic compounds. In the case of organic compounds, second-order rate constants for chlorination vary over 10 orders of magnitude (i.e. <0.1-10(9)M(-1)s(-1)). Oxidation, addition and electrophilic substitution reactions with organic compounds are possible pathways. However, from a kinetic point of view, usually only electrophilic attack is significant. Chlorine reactivity limited to particular sites (mainly amines, reduced sulfur moieties or activated aromatic systems) is commonly observed during chlorination processes and small modifications in the parent compound's structure are expected for the primary attack. Linear structure activity relationships can be used to make predictions/estimates of the reactivity of functional groups based on structural analogy. Furthermore, comparison of chlorine to ozone reactivity towards aromatic compounds (electrophilic attack) shows a good correlation, with chlorine rate constants being about four orders of magnitude smaller than those for ozone. PMID- 17915285 TI - Extending and calibrating a mechanistic hindered and compression settling model for activated sludge using in-depth batch experiments. AB - Currently, no mechanistic model is available in wastewater industry that can accurately describe the batch settling behaviour of activated sludge. Such a model, which is based on the fundamental mass and force balances for water and solids, is extended and applied in this work and excellently describes batch settling experiments for sludges originating from two different wastewater treatment plants. The mechanistic model contains a Kynch batch density function f(bk) (hindered settling) and an effective solids stress function sigma e (compression). Initial settling velocities were obtained from detailed spatio temporal dynamic solids concentration profiles measured with the aid of a radiotracer [De Clercq, J., Jacobs, J., Kinnear, D.J., Nopens, I., Dierckx, R.A., Defrancq, J., Vanrolleghem, P.A., 2005. Detailed spatio-temporal solids concentration profiling during batch settling of activated sludge using a radiotracer. Water Res. 39(10), 2125-2135]. Moreover, inverse modelling calculations were performed using the same data set. Both calculations showed that (1) the power function of Cole gave acceptable results and (2) a single effective solids stress function could be found when a time-dependent compression solids concentration C C was considered. This compression solids concentration is found just below the sludge blanket and is readily calculated from the solids concentration profiles. Given these time-evolutions, the effective solids stress values exhibit a uniform logarithmic relationship with the difference between the solids concentration and the compression solids concentration. The descriptive power of the model indicates a good potential for wider applicability of the model. PMID- 17915286 TI - Predictions of nitrate diffusion in sediment using horizontal attenuated total reflection (HATR) by Fourier transform infrared (FTIR) spectrometry. AB - Using a novel and simple method based on horizontal attenuated total reflection (HATR) by Fourier transform infrared (FTIR) spectrometry, the effective diffusion coefficient, De, of nitrate in a contaminated anthropogenic sediment was estimated as 7.34 x 10(-6)cm2 s(-1). This method, which requires as little as 1 mL of sediment sample, was able to measure the De of a chemical species with a reproducibility of +/-3% in about 5h. Based on this De and a pre-determined nitrate reduction rate, the profiles of nitrate concentration in two sediment columns were satisfactorily predicted from a mathematical model. Results showed that the profile in this aged sediment depended mainly on the diffusion of nitrate and, only to a much lesser degree, the rate of nitrate reduction. Measurements in 55 anthropogenic sediment samples collected from five locations and various depths of a contaminated site further showed that the De of nitrate increased linearly with the water content of the sediment, but decreased with the sediment density. The technique demonstrated in this study shall be applicable for the risk assessment of toxic pollutants in contaminated sediments, and for planning the spatial and time intervals of nitrate injection strategy in bioremediation. PMID- 17915287 TI - Assessment of in situ degradation of chlorinated ethenes and bacterial community structure in a complex contaminated groundwater system. AB - The occurrence of in situ degradation of chlorinated ethenes was investigated using an integrated approach in a complex groundwater system consisting of several geological units. The assessment of hydrogeochemistry and chlorinated ethenes distribution using principal component analysis (PCA) in combination with carbon stable isotope analysis revealed that chlorinated ethenes were subjected to substantial biodegradation. Shifts in isotopic values up to 20.4 per thousand, 13.9 per thousand, 20.1 per thousand and 31.4 per thousand were observed between geological units for tetrachloroethene (PCE), trichloroethene (TCE), cis dichloroethene (cDCE) and vinyl chloride (VC), respectively. The use of specific biomarkers (16S rRNA gene) indicated the presence of Dehalococcoides sp. DNA in 20 of the 33 evaluated samples. In parallel, the analysis of changes in the bacterial community composition in the aquifers using canonical correspondence analysis (CCA) indicated the predominant influence of the chlorinated ethene concentrations (56.3% of the variance, P=0.005). The integrated approach may open new prospects for the assessment of spatial and temporal functioning of bioattenuation in contaminated groundwater systems. PMID- 17915288 TI - Potential targeted therapy options in the management of basal cell subtype of breast carcinoma. PMID- 17915289 TI - Tectonics, orbital forcing, global climate change, and human evolution in Africa: introduction to the African paleoclimate special volume. AB - The late Cenozoic climate of Africa is a critical component for understanding human evolution. African climate is controlled by major tectonic changes, global climate transitions, and local variations in orbital forcing. We introduce the special African Paleoclimate Issue of the Journal of Human Evolution by providing a background for and synthesis of the latest work relating to the environmental context for human evolution. Records presented in this special issue suggest that the regional tectonics, appearance of C(4) plants in East Africa, and late Cenozoic global cooling combined to produce a long-term drying trend in East Africa. Of particular importance is the uplift associated with the East African Rift Valley formation, which altered wind flow patterns from a more zonal to more meridinal direction. Results in this volume suggest a marked difference in the climate history of southern and eastern Africa, though both are clearly influenced by the major global climate thresholds crossed in the last 3 million years. Papers in this volume present lake, speleothem, and marine paleoclimate records showing that the East African long-term drying trend is punctuated by episodes of short, alternating periods of extreme wetness and aridity. These periods of extreme climate variability are characterized by the precession-forced appearance and disappearance of large, deep lakes in the East African Rift Valley and paralleled by low and high wind-driven dust loads reaching the adjacent ocean basins. Dating of these records show that over the last 3 million years such periods only occur at the times of major global climatic transitions, such as the intensification of Northern Hemisphere Glaciation (2.7-2.5 Ma), intensification of the Walker Circulation (1.9-1.7 Ma), and the Mid-Pleistocene Revolution (1-0.7 Ma). Authors in this volume suggest this onset occurs as high latitude forcing in both Hemispheres compresses the Intertropical Convergence Zone so that East Africa becomes locally sensitive to precessional forcing, resulting in rapid shifts from wet to dry conditions. These periods of extreme climate variability may have provided a catalyst for evolutionary change and driven key speciation and dispersal events amongst mammals and hominins in Africa. In particular, hominin species seem to differentially originate and go extinct during periods of extreme climate variability. Results presented in this volume may represent the basis of a new theory of early human evolution in Africa. PMID- 17915290 TI - Mammalian body size changes and Plio-Pleistocene environmental shifts: implications for understanding hominin evolution in eastern and southern Africa. AB - This study examines geographic and temporal variation in three mammalian taxa co occurring in eastern and southern Africa. The selected taxa-the spotted hyena (Crocuta crocuta), the plains zebra (Equus burchellii), and the impala (Aepyceros melampus)--are geographically widespread in modern times and are abundant in eastern and southern African Plio-Pleistocene fossil sites. Craniodental measurements of modern conspecifics from known geographic locations are compared using multivariate statistical methods to discern patterns of modern geographic variation within taxa. Modern and fossil samples are statistically compared to assess the nature and extent of inferred shifts in body size, both between modern samples and through time in each region. These results indicate that modern spotted hyenas and plains zebras exhibit mainly size variation between regions, with southern African samples possessing statistically larger craniodental metrics than eastern African samples. Comparison of fossil and modern samples reveals that the fossil assemblages do not show the same pattern of geographic variation. Significant temporal changes are more numerous between fossil and modern eastern African samples, and these changes are not mirrored by similar changes in the southern African samples. The changes experienced by taxa in eastern Africa appear to have been more extreme and wide-ranging than those in southern Africa, a presumed refugium. This result accords well with genetic studies of several large mammal species and paleoenvironmental studies suggesting that eastern African localized environments were more affected by tectonism and volcanism than were those in southern Africa. This study suggests that different evolutionary scenarios may have existed within Africa during the Plio Pleistocene, but that both regions played unique and complementary roles in the evolution of African hominins and the broader faunal community. PMID- 17915291 TI - Assessing the impact of changes in landuse and management practices on the diffuse pollution and retention of nitrate in a riparian floodplain. AB - In many European lowland rivers and riparian floodplains diffuse nutrient pollution is causing a major risk for the surface waters and groundwater to not achieve a good status as demanded by the European Water Framework Directive. In order to delimit the impact of diffuse nutrient pollution substantial and often controversial changes in landuse and management are under discussion. In this study we investigate the impact of two complex scenarios considering changes in landuse and land management practices on the nitrate loads of a typical lowland stream and the riparian groundwater in the North German Plains. Therefore the impacts of both scenarios on the nitrate dynamics, the attenuation efficiency and the nitrate exchange between groundwater and surface water were investigated for a 998.1 km(2) riparian floodplain of the Lower and Central Havel River and compared with the current conditions. Both scenarios target a substantial improvement of the ecological conditions and the water quality in the research area but promote different typical riparian landscape functions and consider a different grade of economical and legal feasibility of the proposed measures. Scenario 1 focuses on the optimisation of conservation measures for all natural resources of the riparian floodplain, scenario 2 considers measures in order to restore a good status of the water bodies mainly. The IWAN model was setup for the simulation of water balance and nitrate dynamics of the floodplain for a perennial simulation period of the current landuse and management conditions and of the scenario assumptions. The proposed landuse and management changes result in reduced rates of nitrate leaching from the root zone into the riparian groundwater (85% for scenario 1, 43% for scenario 2). The net contributions of nitrate from the floodplain can be reduced substantially for both scenarios. In case of scenario 2 a decrease by 70% can be obtained. For scenario 1 the nitrate exfiltration rates to the river drop even below the infiltration rates from the river, the riparian floodplain in that scenario represents a net sink for river derived nitrate. As the nitrate contributions from the investigated riparian floodplain represent only a small proportion of the total nitrate loads within the river (1% p.a.) the overall impact of the scenario measures on the nitrate loads at the river outlet remains small. However, during the ecologically most sensitive summer periods under current conditions nitrate contributions from the riparian groundwater of the Lower and Central Havel River (which covers only 5% of the area of the Havel catchment) represent more than 20% of the river loads. By the implementation of the investigated landuse changes within the research area the groundwater derived nitrate contributions could be halved to only 10% during summer baseflow conditions. PMID- 17915292 TI - Chlorinated organic pesticides in marketed food: Barcelona, 2001-06. AB - This paper reports concentration levels of 22 chlorinated organic compounds (both primary compounds and metabolites) in food marketed in the city of Barcelona (Catalonia, Spain) in 2001-06. Samples included meat products, fish and seafood, eggs, milk and dairy, vegetal oils, cereal products and derivates, vegetables, fresh fruits, dry fruits, spices, formula and baby food, tea and wine. Levels of chlorinated organic compounds were determined by gas chromatography with selective detectors: electron capture (ECD), flame photometric (FPD) and confirmation with mass-spectrometry. Chlorinated organic pesticides were detected in 7 of the 1,484 samples analyzed in the 2001-06 period (0.5%): 1 dairy product, 1 fruit, 1 olive oil and 4 vegetables. Specific pesticides detected are lindane and endosulfan alpha, beta or sulphate. A decrease in both the proportion of samples with detectable residues and in the variety of chlorinated pesticides found is visible when comparing these results with those of the previous 1989 2000 period. These results suggest the gradual disappearance of regulated chlorinated organic pesticides as a consequence of the growing worldwide implementation of current regulatory agreements. PMID- 17915293 TI - Letter to the editor re: Datta et al., 2006. Arsenic biogeochemistry and human health risk assessment in organo-arsenical pesticide-applied acidic and alkaline soils: an incubation study. PMID- 17915294 TI - Drinking water consumption patterns in British Columbia: an investigation of associations with demographic factors and acute gastrointestinal illness. AB - A cross-sectional telephone survey was performed in the province of British Columbia, Canada, to investigate drinking water consumption patterns and their associations with various demographic characteristics and acute gastrointestinal illness (AGI). Water consumption included plain water and water used in the preparation of cold beverages. The median amount of water consumed daily was four 250 mL servings (1.0 L), although responses were highly variable (0 to 9.0 L). Alternative water use was common: bottled water was the primary source of drinking water (i.e. >or=75% of the total daily water intake) for 23% of respondents and 47% of households used in-home water treatment methods. Approximately 10% of respondents reported an episode of AGI (vomiting or diarrhea) in the previous 4-week period. Such illness was associated with age (continuous variable in years, OR=-0.98), sex (male vs. female, OR=0.8) and the amount of water consumed (continuous variable in 250-mL servings, OR=1.06); however, a causal relationship with water consumption cannot be established based on this study alone. Overall, the associations of drinking water patterns with age, sex, education, and household income serve as important reminders to researchers and public health professionals of the non-uniform nature of drinking water consumption, and indicate potential differences in exposure to waterborne hazards in this population. PMID- 17915295 TI - Thrombin generation determined by calibrated automated thrombography (CAT) in pediatric patients with congenital heart disease. AB - INTRODUCTION: Thrombin generation was studied in pediatric patients with congenital heart disease (CHD) undergoing cardiac surgery using the calibrated automated thrombography (CAT) in terms of the lag time until the onset of thrombin formation, time to thrombin peak maximum (TTP), endogenous thrombin potential (ETP), and thrombin peak height. The possible suitability to determine the coagulation status of these patients was investigated. MATERIALS AND METHODS: CAT data of 40 patients with CHD (age range from newborn to 18 years) were compared to data using standard coagulation parameters such as prothrombin (FII), antithrombin (AT), tissue factor pathway inhibitor (TFPI), prothrombin fragment 1.2 (F 1.2), thrombin-antithrombin (TAT), activated partial thromboplastin time (aPTT), and prothrombin time (PT). RESULTS: A significant positive correlation was seen between ETP and FII (p<0.01; r=0.369), as well as between peak height and F II (p<0.01; r=0.483). A significant negative correlation was seen between ETP and TFPI values (p<0.05; r=-0.225) while no significant correlation was seen between peak height and TFPI. A significant negative correlation was seen between F 1.2 generation and ETP (p<0.05; r=-0.254) and between F 1.2 generation and peak height (p<0.05; r=-0.236). No correlation was seen between AT and ETP or peak. CONCLUSIONS: Our data indicate that CAT is a good global test reflecting procoagulatory and inhibitory factors of the hemostatic system in pediatric patients with CHD. PMID- 17915296 TI - TAFI deficiency in liver cirrhosis: relation with plasma fibrinolysis and survival. AB - INTRODUCTION: TAFI (thrombin-activatable fibrinolysis inhibitor) is a potent anti fibrinolytic and anti-inflammatory factor of liver origin. It is markedly reduced in liver cirrhosis but its effect on fibrinolysis remains controversial and no data are available on its prognostic value. We evaluated the relationship of TAFI level with plasma fibrinolysis and survival in cirrhotic patients. PATIENTS AND METHODS: Sixty-five patients with liver cirrhosis were studied. TAFI antigen, plasma fibrinolysis and other laboratory variables were assayed at study entry and their association with mortality was assessed during a 3-year follow-up. RESULTS: TAFI level and fibrinolysis time were markedly reduced in liver cirrhosis as compared to healthy subjects (p<0.0001) and TAFI deficiency was strongly correlated with fibrinolysis time (p=0.0002). TAFI level at entry, but not fibrinolysis time, was significantly lower in non-survivors (n=25) than in survivors (n=40, p=0.0001). By Cox regression analysis, after adjustment for possible confounding factors, TAFI, but not fibrinolysis time, was identified as an independent predictor of mortality. TAFI assay, moreover, showed a clinically relevant accuracy in assessing patients' survival (ROC curve analysis, p<0.0001) achieving a sensitivity of 92%, a specificity of 55%, and a negative predictive value of 91.7%. CONCLUSIONS: Our data indicate that TAFI deficiency in liver cirrhosis is associated with enhanced plasma fibrinolysis. Moreover, they suggest that TAFI, but not fibrinolysis time, is a strong predictor of survival and thus TAFI assay might prove useful to select candidates for liver transplantation. PMID- 17915297 TI - Electrophysiological modelling of pulmonary artery smooth muscle cells in the rabbits--special consideration to the generation of hypoxic pulmonary vasoconstriction. AB - In vascular smooth muscle cells, it has been suggested that membrane potential is an important component that initiates contraction. We developed a mathematical model to elucidate the quantitative contributions of major ion currents [a voltage-gated L-type Ca2+ current (ICaL), a voltage-sensitive K+ current (IKV), a Ca2+-activated K+ current (IKCa) and a nonselective cation current (INSC)] to membrane potential. In order to typify the diverse nature of pulmonary artery smooth muscle cells (PASMCs), we introduced parameters that are not fixed (variable parameters). The population of cells with different parameters was constructed and the cells that have the electrophysiological properties of PASMCs were selected. The contributions of each membrane current were investigated by sensitivity analysis and modification of the current parameters. Consequently, IKV and INSC were found to be the most important currents that affect the membrane potential. The occurrence of depolarisation in hypoxic pulmonary vasoconstriction (HPV) was also examined. In hypoxia, IKV and IKCa were reduced, but the consequent depolarisation in simulation was not enough to initiate contractions. If we add an increase of INSC (2.5-fold), the calculated membrane potential was enough to induce contraction. From the results, we conclude that the balance of various ion channel activities determines the resting membrane potential of PASMCs and our model was successful in explaining the depolarisation in HPV. Therefore, this model can be a powerful tool to investigate the various electrical properties of PASMCs in both normal and pathological conditions. PMID- 17915298 TI - The canine virtual ventricular wall: a platform for dissecting pharmacological effects on propagation and arrhythmogenesis. AB - We have constructed computational models of canine ventricular cells and tissues, ultimately combining detailed tissue architecture and heterogeneous transmural electrophysiology. The heterogeneity is introduced by modifying the Hund-Rudy canine cell model in order to reproduce experimentally reported electrophysiological properties of endocardial, midmyocardial (M) and epicardial cells. These models are validated against experimental data for individual ionic current and action potential characteristics, and their rate dependencies. 1D and 3D heterogeneous virtual tissues are constructed, with detailed tissue architecture (anisotropy and orthotropy, due to fibre orientation and sheet structure) of the left ventricular wall wedge extracted from a diffusion tensor imaging data set. The models are used to study the effects of tissue heterogeneity and class III drugs on transmural propagation and tissue vulnerability to re-entry. We have determined relationships between the transmural dispersion of action potential duration (APD) and the vulnerable window in the 1D virtual ventricular wall, and demonstrated how changes in the transmural heterogeneity, and hence tissue vulnerability, can lead to generation of re-entry in the 3D ventricular wedge. Two class III drugs with opposite qualitative effects on transmural APD heterogeneity are considered: d-sotalol that increases transmural APD dispersion, and amiodarone that decreases it. Simulations with the 1D virtual ventricular wall show that under d-sotalol conditions the vulnerable window is substantially wider compared to amiodarone conditions, primarily in the epicardial region where unidirectional conduction block persists until the adjacent M cells are fully repolarised. Further simulations with the 3D ventricular wedge have shown that ectopic stimulation of the epicardial region results in generation of sustained re-entry under d-sotalol conditions, but not under amiodarone conditions or in control. Again, APD increase in M cells was identified as the major contributor to tissue vulnerability--re-entry was initiated primarily due to ectopic excitation propagating around the unidirectional conduction block in the M cell region. This suggests an electrophysiological mechanism for the anti- and proarrhythmic effects of the class III drugs: the relative safety of amiodarone in comparison to d-sotalol can be explained by relatively low transmural APD dispersion, and hence, a narrow vulnerable window and low probability of re-entry in the tissue. PMID- 17915300 TI - Gemcitabine and pegylated liposomal doxorubicin alternating with cisplatin plus cyclophosphamide in platinum refractory/resistant, paclitaxel-pretreated, ovarian carcinoma. AB - OBJECTIVES: This phase II study conducted to investigate the efficacy and toxicity of the combination of gemcitabine (GEM) and pegylated liposomal doxorubicin (LDOX) alternating with cisplatin (CDDP) and cyclophosphamide (CTX) in platinum-resistant/refractory, paclitaxel pretreated epithelial ovarian cancer (EOC). PATIENTS AND METHODS: Forty-eight patients with CDDP-resistant/refractory and paclitaxel pretreated patients were treated with 8 cycles of GEM 800 mg/m2 days 1 and 8 and LDOX 30 mg/m2 day 1, alternating with CDDP 60 mg/m2 and CTX 600 mg/m2 every 3 weeks. RESULTS: Objective responses were observed in 37.5% of patients (4 complete and 11 partial responses) with measurable disease (n=40). CA125 response occurred in 30 (71.4%) of patients with elevated CA125 (n=42). After a median follow-up of 23 months, the median progression-free survival (PFS) was 6.9 months (95% confidence interval, CI: 5.2-8.5), while the median overall survival (OS) was 18.2 months (95% CI: 12.7-23.6). A progression-free interval (PFI) of 0-3 months was associated with lower objective responses (10% versus 46.6%, p=0.06). Chemotherapy was well tolerated. The most frequent toxicities were myelosuppression, neurotoxicity, nephrotoxicity, nausea/vomiting, fatigue and palmar-plantar erythrodysesthesia (PPE). Overall 31 (65%) patients received G CSF and 13 (27%) antibiotics because of neutropenia and/or febrile neutropenia. CONCLUSION: This alternating combination chemotherapy is feasible for patients with platinum-resistant EOC and is associated with encouraging outcomes and a favorable toxicity profile. PMID- 17915299 TI - The role of transmural ventricular heterogeneities in cardiac vulnerability to electric shocks. AB - Transmural electrophysiological heterogeneities have been shown to contribute to arrhythmia induction in the heart; however, their role in defibrillation failure has never been examined. The goal of this study is to investigate how transmural heterogeneities in ionic currents and gap-junctional coupling contribute to arrhythmia generation following defibrillation strength shocks. This study used a 3D anatomically realistic bidomain model of the rabbit ventricles. Transmural heterogeneity in ionic currents and reduced sub-epicardial intercellular coupling were incorporated based on experimental data. The ventricles were paced apically, and truncated-exponential monophasic shocks of varying strength and timing were applied via large external electrodes. Simulations demonstrate that inclusion of transmural heterogeneity in ionic currents results in an increase in vulnerability to shocks, reflected in the increased upper limit of vulnerability, ULV, and the enlarged vulnerable window, VW. These changes in vulnerability stem from increased post-shock dispersion in repolarisation as it increases the likelihood of establishment of re-entrant circuits. In contrast, reduced sub epicardial coupling results in decrease in both ULV and VW. This decrease is caused by altered virtual electrode polarisation around the region of sub epicardal uncoupling, and specifically, by the increase in (1) the amount of positively polarised myocardium at shock-end and (2) the spatial extent of post shock wavefronts. PMID- 17915301 TI - Estrogens participate in the antidepressant-like effect of desipramine and fluoxetine in male rats. AB - In male rats, the antidepressant-like effect of fluoxetine (FLX) and desipramine (DMI) in the forced swimming test (FST) is reduced by orchidectomy and partially restored by testosterone (T). It is unknown if this modulation of T is produced by its estrogenic metabolites. The objectives of this study were to evaluate if the aromatase inhibitor, formestane, interferes with the antidepressant-like effect of DMI and FLX in intact male rats, and to analyze if 17beta-estradiol (E2) modifies the FST and interacts with the antidepressants in orchidectomized (Orx) males. Intact males received DMI (1.25-5.0 mg/kg) and FLX (2.5-10 mg/kg) alone or in combination with formestane (17.5 mg/kg). Orx rats received E2 (5, 10, 20 and 40 microg/rat) or the combination of E2 [at sub-threshold (5 microg/rat) and optimal (10 microg/rat) doses] plus sub-effective doses of DMI (2.5 mg/kg) or FLX (10 mg/kg). Serum testosterone and estradiol levels were measured in intact-control and -formestane treated animals as well as in castrated males replaced with various doses of E2. Formestane in intact males lacked of an action in the FST, but cancelled the antidepressant-like effect of DMI and FLX. E2 at the supra-physiological doses of 10 and 20 microg/rat produced antidepressant-like effects. E2 at 5 microg/rat (that re-established the levels of this hormone to physiological levels) and at 10 microg/rat restored the antidepressant-like action of DMI and FLX in Orx rats. It was concluded that estrogens participate in the antidepressant-like effect of DMI and FLX in the FST. PMID- 17915303 TI - Nitric oxide-mediated anxiolytic-like and antidepressant-like effects in animal models of anxiety and depression. AB - The effects of microinjection of the nitric oxide (NO) precursor l-arginine (l Arg), the NO synthase (NOS) inhibitors N-methyl-l-arginine (l-NAME) and 7 nitroindazole (7-NI), and the cyclic guanosine 3',5'-monophosphate (cGMP) analog 8-Br-cGMP into the dorsal raphe nucleus (DRN) were assessed in rats using the elevated plus maze (EPM) and the forced swim test (FST). l-Arg (100 and 200 nmol) produced an anxiolytic-like effect in the EPM. 8-Br-cGMP (25 and 50 nmol) dose dependently increased locomotor activity. In the FST, antidepressant-like effects were produced by l-Arg (50 and 100 nmol) and 8-Br-cGMP (12.5 and 25 nmol). Dual effects were observed with NOS inhibitors l-NAME and 7-NI in both the EPM and FST. While low doses of l-NAME (25 nmol) or 7-NI (1 nmol) induced a selective increase in EPM open arm exploration and a decrease in immobility time in the FST, high doses (l-NAME 400 nmol, 7-NI 10 nmol) decreased locomotor activity. These results show that interference with NO-mediated neurotransmission in the DRN induced significant and complex motor and emotional effects. Further studies are needed to elucidate the mechanisms involved in these effects. PMID- 17915302 TI - Repeated administration of nicotine attenuates the development of morphine tolerance and dependence in mice. AB - Clinical use of morphine in pain management is a controversial issue. Both nicotine and morphine are widely abused. So, investigating the interaction between nicotinic and opioid receptors is of great interest to both basic mechanistic and clinical view. We investigated the influence of repeated administration of nicotine on the development of morphine tolerance and dependence. Adult male albino mice were rendered dependent on morphine by subcutaneous (s.c.) injections three times daily for 3 days. Repeated intraperitoneal (i.p.) injection of nicotine (0.001-2 mg/kg) or saline (1 ml/kg) was performed 15 min prior to each morphine injection. Maximal possible effect (MPE%) of morphine (50 mg/kg; s.c.) was used on the fourth day as an index for the development of tolerance. Likewise, to assess the occurrence of dependence in drug-treated mice, naloxone (5 mg/kg; i.p.) was injected 2 h after the last dose of morphine. Repeated nicotine administration significantly attenuated the development of tolerance in a dose-dependent manner whereas it significantly decreased withdrawal jumping behavior in a biphasic profile (V-shape) manner. Furthermore, the central nicotinic receptor antagonist mecamylamine (0.01-0.1 mg/kg; i.p.) neither the peripheral nicotinic receptor antagonist hexamethonium (0.01 and 0.1 mg/kg; i.p.) nor the muscarinic receptor antagonist atropine (2.5 10 mg/kg; i.p.), dose-dependently antagonized both the inhibition of withdrawal jumping as well as increase in MPE% which was produced by repeated nicotine administration (0.1 mg/kg; i.p.). On the other hand, 3 days of solely nicotine treatment resulted in significant jumping behavior precipitated by naloxone after single morphine injection on the test day. The data suggests that the inhibitory effect of nicotine on the morphine tolerance and dependence is mediated by central nicotinic receptors and there is a cross-dependence between nicotine and morphine. PMID- 17915304 TI - Gene expression and development of mouse zygotes following droplet vitrification. AB - The concept of ultra-rapid vitrification has emerged in recent years; the accelerated cooling rate reduced injury attributed to cryopreservation and improved post-freezing developmental competence of vitrified oocytes and embryos. The objectives of the present study were to develop a simple and effective ultra rapid vitrification method (droplet vitrification) and evaluate its effects on post-thaw development and apoptosis-related gene expression in mouse zygotes. Presumptive zygotes were equilibrated for 3 min in equilibration medium and washed 3 times in vitrification solution. A drop (5 microL) of vitrification solution containing 10-12 embryos was placed directly onto surface of liquid nitrogen, with additional liquid nitrogen poured over the drop. For thawing and cryoprotectant removal, vitrified drops were put into dilution medium for 3 min, followed by M2 medium for 5 min. Although cleavage rate did not differ significantly among the control (90.8+/-2.8%; mean+/-S.E.M.), toxicity control (83.5+/-3.2%), and vitrified (86.2+/-3.1%) zygotes, rates of blastocyst and hatched blastocyst formation were lower (P<0.01) in vitrified zygotes (49.7+/ 4.7% and 36.0+/-4.7%) and toxicity controls (47.3+/-4.6% and 40.3+/-4.6%) compared with controls (65.5+/-4.1% and 54.2+/-4.3%). Exposure of zygotes to vitrification solution, as well as the vitrification process, down-regulated the expression of Bax, Bcl2, and p53 genes in blastocysts. Although droplet vitrification was efficient and easy, it altered the transcriptional activities of Bax, Bcl2, and p53 genes in vitrified embryos, indicating a strong relationship between reduced developmental competence and the altered transcriptional activities of these genes. PMID- 17915305 TI - Ultrasonographic imaging of the reproductive tract and surgical recovery of oocytes in Cebus apella (capuchin monkeys). AB - Two-dimensional real-time and Doppler ultrasonography are valuable non-invasive methods to assess reproductive anatomy and physiology. In adult, postpubertal female Cebus apella (capuchin monkeys), the objectives were to determine (1) uterine and ovarian dimensions, ovarian follicular dynamics, day of ovulation, and arterial blood flow of uterus and utero-ovarian ligament during the follicular phase of the menstrual cycle and (2) the number of oocytes aspirated from antral follicles at laparotomy. Based on two-dimensional, transabdominal B mode ultrasonography, mean (+/- S.E.M.) length, height, width, and volume of the uterus were 17.9+/-0.4, 12.4+/-0.3, 13.6+/-0.3 mm, and 1.55+/-0.08 mL, respectively, and of the ovary were 13.4+/-0.2, 8.2+/-0.1, 7.7+/-0.1 mm, and 4.5+/-0.2 mL. Ovarian follicles were monitored for 6 days before ovulation, which occurred on day 9.3+/-0.5 (range, days 7-11; day 1=start of menses), with 10 of 12 ovulations in the right ovary. Diameter and volume of the preovulatory follicle were 10.1+/-0.2 mm and 0.55+/-0.03 mL (on the estimated day of ovulation) and of the CL were 8.1+/-0.4 mm and 0.3+/-0.05 mL. Resistivity and pulsatility indices were 0.86+/-0.02 and 2.15+/-0.11 for uterine arteries, and were 0.69+/-0.04 and 1.63+/-0.15 for the utero-ovarian ligament (UOL) artery; just prior to ovulation, both indices peaked (P<0.05) in the uterine artery ipsilateral to the side of ovulation, but both reached a nadir (P<0.05) in the UOL artery. In the absence of ovarian stimulation, 31 oocytes (diameter, 137+/-10 microm) were aspirated (average of 2 oocytes/(female attempt)) on days 5, 7, and 9. In conclusion, transabdominal ultrasonography facilitated assessment of reproductive anatomy and physiology in C. apella adult females. Resistance and pulsatility indices of uterine and UOL arteries changed near the time of ovulation. Dominant follicles were easiest to aspirate at 8-9 mm in diameter ( approximately day 9), with intact cumulus-oocyte complexes recovered from ovarian follicles 2-9 mm in diameter. PMID- 17915306 TI - Chronological appearance of spontaneous and induced apoptosis during preimplantation development of rabbit and mouse embryos. AB - This study was undertaken to obtain specific information on the characteristics of spontaneous and induced apoptosis during preimplantation development of rabbit in vivo and in vitro developed embryos and mouse in vitro embryos. After reaching appropriate developmental stages, embryos were transferred into culture media with or without apoptotic inductor (actinomycin D 500 ng/mL) and cultured for 10 h. The identification of apoptotic cells was based on morphological assessment of nuclei and on detection of specific DNA degradation, phosphatidylserine redistribution and active caspase-3 under fluorescence microscope. Our experiments proved that apoptosis is a frequent physiological event occurring during normal preimplantation development. A high number of untreated rabbit and mouse blastocysts contained at least one apoptotic cell. Rabbit embryos showed a lower incidence of spontaneous apoptosis. Treated blastocysts of both species responded to the presence of apoptotic inductor by significant decrease in the average number of blastomeres and significant increase in the incidence of apoptotic cell death. The occurrence of spontaneous apoptosis during earlier preimplantation development was sporadic and its presence was observed only at stages following embryonic genome activation (at 4-cell stage and later in mouse, at 16-cell and morula stage in rabbit). The susceptibility of embryos at early stages to the apoptotic inductor was much lower. The presence of actinomycin D did not increase the incidence of apoptotic embryos or apoptotic cells. Nevertheless, it slowed down embryo growth and triggered earlier appearance of some apoptotic features (at the 6-cell stage in rabbit). The results show that the occurrence of both spontaneous and induced apoptosis in preimplantation embryos is stage- and species-specific. PMID- 17915307 TI - Unusual and abnormal canine estrous cycles. AB - Preovulatory serum progesterone concentrations are used to estimate the day of LH peak (day 0), not only to accurately time insemination and predict parturition, but to identify abnormal or unusual estrous cycles due to ovarian dysfunction. Early identification of these disorders is of therapeutic and economic importance. This review discusses anovulation, slow preovulatory progesterone rise, "split heat", insufficient luteal phase, and persistent estrus in the bitch. Some of these were temporary dysfunctions; with appropriate breeding management, pregnancy can be achieved. However, in other cases, these were signs of severe, permanent ovarian dysfunction associated with infertility, with potentially lethal sequelae. PMID- 17915308 TI - Application of zero-length cross-linking to form lysozyme, horseradish peroxidase and lysozyme-peroxidase dimers: Activity and stability. AB - A facile method for the formation of covalent bonds between protein molecules is zero-length cross-linking. This method enables the formation of cross-links without use of any chemical reagents. Here, the cross-linking is performed for lysozyme, peroxidase (a glycoprotein) and between lysozyme-peroxidase by the method of Simons et al. [B.L. Simons, M.C. King, T. Cyr, M.A. Hefford, H. Kaplan, Covalent cross-linking of protein without chemical reagents, Protein Sci. 2002, 11, 1558-1564]. Approximately one-third of the total lysozyme becomes cross linked and the dimer form was the major product for both enzymes. This modification induced some changes in the kinetic properties of the dimer peroxidase, as evident by two-fold increasing of V(max) compared to the monomer but the enzymatic activity of cross-linked lysozyme dimer was the same as monomer. The activity of lysozyme dimer remained constant up to 10min at 80 degrees C, while peroxidase activity of both monomer and dimer began to decrease after heating. The structural changes of the enzymes were investigated by circular dichroism and intrinsic fluorescence techniques. Near UV result showed lysozyme possess a compact structure in the dimer form but disruption of tertiary structure of peroxidase dimer was observed. Also conformational changes were detected and discussed by intrinsic fluorescence experiments. Effect of several metals in the formation of lysozyme dimer showed that Co(2+) is the most effective one but its effect was marginal. At the end formation of heterogeneous dimer, peroxidase-lysozyme, was achieved using this method. PMID- 17915309 TI - Characterisation of a soft elastomer poly(glycerol sebacate) designed to match the mechanical properties of myocardial tissue. AB - The myocardial tissue lacks significant intrinsic regenerative capability to replace the lost cells. Therefore, the heart is a major target of research within the field of tissue engineering, which aims to replace infarcted myocardium and enhance cardiac function. The primary objective of this work was to develop a biocompatible, degradable and superelastic heart patch from poly(glycerol sebacate) (PGS). PGS was synthesised at 110, 120 and 130 degrees C by polycondensation of glycerol and sebacic acid with a mole ratio of 1:1. The investigation was focused on the mechanical and biodegrading behaviours of the developed PGS. PGS materials synthesised at 110, 120 and 130 degrees C have Young's moduli of 0.056, 0.22 and 1.2 MPa, respectively, which satisfy the mechanical requirements on the materials applied for the heart patch and 3D myocardial tissue engineering construction. Degradation assessment in phosphate buffered saline and Knockout DMEM culture medium has demonstrated that the PGS has a wide range of degradability, from being degradable in a couple of weeks to being nearly inert. The matching of physical characteristics to those of the heart, the ability to fine tune degradation rates in biologically relevant media and initial data showing biocompatibility indicate that this material has promise for cardiac tissue engineering applications. PMID- 17915310 TI - Thermoplastic biodegradable polyurethanes: the effect of chain extender structure on properties and in-vitro degradation. AB - Biodegradable polyurethanes are typically prepared from polyester polyols, aliphatic diisocyanates and chain extenders. We have developed a degradable chain extender (DCE) based on dl-lactic acid and ethylene glycol to accelerate hard segment degradation. Three series of polyurethane elastomers were synthesised to investigate the effect of incorporating DCE on synthesis, mechanical and thermal properties and in-vitro degradation. Polyurethane soft segments were based on poly(epsilon-caprolactone) (PCL) polyol. The hard segment was based on either ethyl lysine diisocyanate or hexamethylene diisocyanate in combination with ethylene glycol or DCE. Polyurethanes were characterised by gel permeation chromatography, tensile testing (Instron) and differential scanning calorimetry. Polymer degradation in-vitro (phosphate buffered saline) was tested by measuring mass loss, change in molecular weight and amine concentration in degradation products at three different time points over a 1 year period. Incorporation of DCE did not affect thermal or mechanical properties but had an influence on the in-vitro degradation. All polyurethanes exhibited considerable molecular weight decrease over the test period, and DCE-based polyurethanes showed the highest mass loss. The presence of the DCE and the initial molecular weight of the polyurethane are the key factors responsible for high mass losses. Differential scanning calorimetry, amine group analysis and the observation that mass loss was directly proportional to hard segment weight percentage, strongly supported that the polyurethane hard segment is the most susceptible segment to degradation in these polyurethanes. The PCL-based soft segment appears to undergo little or no degradation under these test conditions. PMID- 17915311 TI - Muscle-derived stem cells for tissue engineering and regenerative therapy. AB - Skeletal muscle has been recognized as an essential source of progenitor or satellite cells, which are primarily responsible for muscle regeneration. Recently, muscle has also been identified as a valuable source of postnatal stem cells that appear to be distinct from satellite cells and possess the ability to differentiate into other cell lineages. These cells, named muscle-derived stem cells, possess a high myogenic capacity and effectively regenerate both skeletal and cardiac muscle. Remarkably, when genetically modified ex vivo to express growth factors, these cells can differentiate into osteogenic and chondrogenic lineages and have been shown to promote the repair of bone and cartilage. Muscle stem cell-based regenerative therapy and tissue engineering using ex vivo gene therapy, are promising approaches for the treatment of various musculoskeletal, cardiovascular, and urological disorders. PMID- 17915312 TI - Hydrolysable core-crosslinked thermosensitive polymeric micelles: synthesis, characterisation and in vivo studies. AB - In this study, core-crosslinked (CCL) biodegradable thermosensitive micelles based on mPEG(5000) and N-(2-hydroxyethyl)methacrylamide)-oligolactates (mPEG-b p(HEMAm-Lac(n))) were synthesised and their properties investigated. Rapidly heating aqueous solutions of partially methacrylated block copolymers to above their critical micelle temperature (CMT), followed by illumination in presence of a photoinitiator yielded almost monodisperse CCL micelles with a size of 68+/-7 nm. Either below the CMT or after addition of sodium dodecyl sulphate, the non crosslinked (NCL) micelles rapidly disintegrated whereas the CCL micelles kept their integrity. NCL micelles fell apart after 5h in pH 7.4 at 37 degrees C as a result of the hydrolysis of lactate side chains, whereas the CCL micelles had a much higher stability and only degraded after cleavage of the ester bonds in the crosslinks. The circulation kinetics and biodistribution of CCL micelles were considerably better than those of NCL micelles, i.e., 58% of the injected dose (ID) of CCL versus 6% of NCL micelles was recovered in the circulation 4h post injection. Furthermore, the liver uptake of the CCL micelles (10% ID) was much lower than that of the NCL micelles (24% ID) 4h after administration, while tumour accumulation was almost 6 times higher for the CCL micelles. Likely, NCL micelles dissociated after i.v. administration and/or were opsonised and captured by macrophages while the dense PEG shell of CCL micelles made them less prone towards opsonisation. The excellent physical stability of these degradable CCL micelles and very favourable biodistribution profile renders them very suitable for drug targeting purposes. PMID- 17915313 TI - Caviomorph placentation as a model for trophoblast invasion. AB - The guinea pig and its relatives are promising candidates as animal models for studying trophoblast invasion. The origin, migration routes and kinetics of invasive trophoblast cells were examined in two caviomorph species. Histology and immunohistochemistry were done on placentas from 38 guinea pigs of days 20-47 and 13 degus of days 25-51 of gestation. BrdU was used as an in vivo marker for proliferation and for tracing of migration routes in the placenta; it was injected 24h to 15 days before collecting the material. In both species extravillous-like trophoblast cells are derived from proliferating stem cell aggregations in the subplacenta, which are comparable to the cell columns in humans. Migration routes and kinetics under in vivo conditions revealed a mean invasive depth of 300-350 microm/day and a mean life span of the extravillous like trophoblast of 30 days. The patterns of trophoblast invasion in caviomorphs are analogous to the situation in humans, suggesting that these rodents are appropriate animal models for the study of the dynamics of trophoblast invasion. PMID- 17915314 TI - Have myeloma cells osteoclast-like activity? Implications into the pathogenesis of myeloma bone disease. PMID- 17915315 TI - Acquired amegakaryocytic thrombocytopenia and pure red cell aplasia associated with an occult large granular lymphocyte leukemia. AB - Acquired amegakaryocytic thrombocytopenia and pure red cell aplasia rarely occur concurrently. We report a case in which these disorders were associated with an occult large granular lymphocyte leukemia. The peripheral blood cytopenias improved after glucocorticoids and intravenous immunoglobulin were administered, and response was maintained with cyclosporine. Large granular lymphocyte leukemia should be suspected in the setting of unexplained bone marrow failure. PMID- 17915316 TI - Lipomembranous panniculitis in a patient with acute leukemia induced by chemotherapy. PMID- 17915317 TI - Have we been wrong about ionizing radiation and chronic lymphocytic leukemia? AB - It is almost axiomatic that chronic lymphocytic leukemia (CLL) is not caused by ionizing radiation. This assumption has been challenged recently by a critical re appraisal of existing data. A recent paper implicated radon exposure in Czech uranium miners as a possible cause of CLL and in this issue of Leukemia Research the first paper examining the incidence of CLL among those exposed to radiation from the accident at the nuclear power plant in Chernobyl is published. It suggests that CLL occurring among the clean-up workers was of a more aggressive form than is normally seen in the community. PMID- 17915318 TI - All-trans-retinoic acid enhances apoptosis induction by tyrosine kinase inhibitors in the eosinophilic leukemia-derived EoL-1 cell line. AB - Imatinib and retinoids induce apoptosis in FIP1L1/PDGFRalpha-positive EoL-1 leukemia cells. Although imatinib induces complete remission in most FIP1L1/PDGFRalpha-positive patients, response to imatinib is sometimes suboptimal. In order to enhance the potency of the molecularly targeted therapy of eosinophilic leukemia, we investigated the effect of retinoids combined with tyrosine kinase inhibitors on EoL-1 cells. We demonstrate that retinoids combined with tyrosine kinase inhibitors lead to enhanced apoptosis induction in EoL-1 cells. Our results suggest that tyrosine kinase inhibitors combined with retinoids may constitute a valuable therapeutic approach for sensitive neoplasias that may display enhanced anti-leukemic potency when compared to single drug treatments. PMID- 17915319 TI - Assessing toxicity and biodegradation of novel, environmentally benign ionic liquids (1-alkoxymethyl-3-hydroxypyridinium chloride, saccharinate and acesulfamates) on cellular and molecular level. AB - Ionic liquids are widely studied as alternative solvents in organic synthesis and catalysis, in electrochemistry and the separation sciences; with their many applications they will soon be produced on an industrial scale. Available toxicological data of ionic liquids have already suggested initial guidelines for the conscious design of safer chemicals. In this study a new group of such redesigned ionic liquids-1-alkoxymethyl-3-hydroxypyridinium cations+acesulphamate, saccharinate and chloride anions-was assayed with respect to their inhibitory activity towards acetylcholinesterase and their cellular toxicity towards the IPC-81 rat promyelocytic leukaemia cell line: the acute biological activity of these compounds is very low. Effective concentrations lie in the millimole range, which is well above possible intracellular concentrations. Only the compounds with the longest alkoxymethyl chain inhibit the enzyme at effective concentrations that are one order of magnitude smaller. No significant differences are observed when the anion compartment in the enzymatic assay is varied. However, the cytotoxicity data show EC(50) for acesulphamates and saccharinates to be higher than the values for the chloride analogues. Also, a slight alkoxymethyl chain length effect on the overall cytotoxicity is discernible. The biodegradability of the 1-alkoxymethyl-3 hydroxypyridinium salts varies from 21% to 72% and depends on the type of anion the cation is linked with. It improves with lengthening alkyl chain, but only in the range from 4 to 11 carbon atoms. PMID- 17915320 TI - Comparative studies on adsorptive removal of phenol by three agro-based carbons: equilibrium and isotherm studies. AB - The present study deals with the adsorption of phenol on carbon-rich black gram husk (BGH), green gram husk (GGH), and rice husk (RH). All these agro-wastes were collected from the local mills and physico-chemical treatments were carried out to improve their adsorption capacity. Batch studies were performed to evaluate the effect of various experimental parameters such as initial pH, contact time, adsorbent dosage, and initial phenol concentration (C(0)) of 100mg/l. Optimum conditions for phenol removal were found to be at pH 5.1 (in the range 2-12), an adsorbent dosage of 0.5 g/l of solution and for an equilibrium time of 6h. Equilibrium isotherms for the adsorption of phenol on BGH, GGH, and RH were analyzed by Freundlich, Langmuir, Temkin, Redlich-Peterson, and Dubinin Radushkevich isotherm models. Redlich-Peterson isotherm was found to be the best representative for phenol-sorption on all the three adsorbents studied. PMID- 17915321 TI - Yeast culture has anti-inflammatory effects and specifically activates NK cells. AB - Yeast culture is widely used in animal feed and has been linked to beneficial effects on animal health and production. This study examined the anti-oxidant and immunomodulating effects of a consumable yeast culture, XP, in vitro. An aqueous extract of XP contained anti-oxidants able to enter living cells and quench free radicals. The XP extract induced an increased expression of CD69 and CD25 on NK and NKT cells, and an increased cytotoxic response to K562 tumor cells. The XP extract amplified ProteinA-induced B cell activation in vitro, as measured by induction of the CD86 antigen on B lymphoblasts in 7-day cultures. The data show an anti-inflammatory effect of the XP extract in conjunction with activation of NK cells and B lymphocytes in vitro. Further in vivo studies are needed to examine the impact of XP in animals with bacterial and viral infections, as well as around the time of vaccination. PMID- 17915322 TI - Endotoxin activity in whole blood measured by neutrophil chemiluminescence is applicable to canine whole blood. AB - The dog is widely used as a translational experimental model studying the host response and new treatments for human endotoxemia. The present study evaluated the applicability of a novel patient-near neutrophil chemiluminescence assay for the measurement of endotoxin activity in human blood when applied to canine blood samples. The assay was observed to be analytically sensitive and specific to endotoxin when tested in vitro, spiked with purified Escherichia coli lipopolysaccharide and live E. coli. The diagnostic sensitivity was sustained during Gram-positive contamination. Finally, it also demonstrated diagnostic potential when able to discriminate dogs with spontaneously occurring endotoxemia from both healthy dogs and diseased dogs without endotoxemia. The rapid patient near assessment of endotoxin activity in canine blood should facilitate future studies on endotoxemia in both spontaneous disease and in experimental settings. PMID- 17915323 TI - Particle-based vaccines for transcutaneous vaccination. AB - Immunization concepts evolve with increasing knowledge of how the immune system works and the development of new vaccination methods. Traditional vaccines are made of live, attenuated, killed or fragmented pathogens. New vaccine strategies can take advantage of particulate compounds--microspheres or nanoparticles--to target antigen-presenting cells better, which must subsequently reach the secondary lymphoid organs, which are the sites of the immune response. The use of the skin as a target organ for vaccine delivery stems from the fact that immature dendritic cells (DCs), which are professional antigen-presenting cells can be found at high density in the epidermis and dermis of human or animal skin. This has led to design various methods of dermal or transcutaneous vaccination. The quality and duration of the humoral and cellular responses to vaccination depend on the appropriate targeting of antigen-presenting cells, of the vaccine dose, route of administration and use of adjuvant. In this review, we will focus on the use of micro- and nano-particles to target the skin antigen-presenting cells and will discuss recent advances in the field of transcutaneous vaccination in animal models and humans. PMID- 17915325 TI - Development of the Observation Scale for Aggressive Behavior (OSAB) for Dutch forensic psychiatric inpatients with an antisocial personality disorder. AB - The Observation Scale for Aggressive Behavior (OSAB) has been developed to evaluate inpatient treatment programs designed to reduce aggressive behavior in Dutch forensic psychiatric patients with an antisocial personality disorder, who are "placed at the disposal of the government". The scale should have the sensitivity to measure changes in the possible determinants of aggressive behavior, such as limited control of displayed negative emotions (irritation, anger or rage) and a general deficiency of social skills. In developing the OSAB 40 items were selected from a pool of 82 and distributed among the following a priori scales: Irritation/anger, Anxiety/gloominess, Aggressive behavior, Antecedent (to aggressive behavior), Sanction (for aggressive behavior) and Social behavior. The internal consistency of these subscales was good, the inter rater reliability was moderate to good, and the test-retest reliability over a two to three week period was moderate to good. The correlation between the subscales Irritation/anger, Anxiety/gloominess, Aggressive behavior, Antecedent, Sanction was substantial and significant, but the anticipated negative correlation between these subscales and the Social behavior subscale could not be shown. Relationships between the corresponding subscales of the OSAB and the FIOS, used to calculate concurrent validity, yielded relatively high correlations. The validity of the various OSAB subscales could be further supported by significant correlations with the PCL-R and by significant but weak correlations with corresponding subscales of the self-report questionnaires. The Observation Scale for Aggressive Behavior (OSAB) seems to measure aggressive behavior in Dutch forensic psychiatric inpatients with an antisocial personality disorder reliably and validly. Contrary to expectations, a negative relationship was not found between aggressive and social behavior in either the OSAB or FIOS, which were used for calculating concurrent validity. PMID- 17915326 TI - Schizophrenia sex offenders: a clinical and epidemiological comparison study. AB - While the incidence of criminal offense in individuals with schizophrenia does not necessarily differ from that of the normal population, in contrast, the rate of sexual crimes among individuals with schizophrenia is relatively low. Due to low numbers, description of sexual criminal behavior among schizophrenia patients remains relatively unexplored. In order to investigate clinical, socio demographic and sexual factors associated with deviant sexual behavior in schizophrenia sex offenders we retrospectively compared a large subgroup of schizophrenia sex offenders with a group of schizophrenia patients incarcerated for other criminal behaviors and a group of sex offenders without schizophrenia. Results indicated significant differences distinguishing schizophrenia sex offenders from schizophrenia non-sex offenders, the former of whom were more likely to be married, employed, non-heterosexual (homosexual and bisexual orientations) and demonstrated less hospitalization, antisocial personality, substance abuse, negative symptoms and overall illness severity. Individuals with schizophrenia had a tendency towards female victims compared to non-schizophrenia sex offenders whose victims tended to be male. Study observations indicate important differences between the 3 subgroups suggesting that the schizophrenia sex offender subpopulation may be a unique subgroup of comorbid conditions. PMID- 17915324 TI - Deficits in facial affect recognition among antisocial populations: a meta analysis. AB - Individuals with disorders marked by antisocial behavior frequently show deficits in recognizing displays of facial affect. Antisociality may be associated with specific deficits in identifying fearful expressions, which would implicate dysfunction in neural structures that subserve fearful expression processing. A meta-analysis of 20 studies was conducted to assess: (a) if antisocial populations show any consistent deficits in recognizing six emotional expressions; (b) beyond any generalized impairment, whether specific fear recognition deficits are apparent; and (c) if deficits in fear recognition are a function of task difficulty. Results show a robust link between antisocial behavior and specific deficits in recognizing fearful expressions. This impairment cannot be attributed solely to task difficulty. These results suggest dysfunction among antisocial individuals in specified neural substrates, namely the amygdala, involved in processing fearful facial affect. PMID- 17915327 TI - Occurrence of phthalate esters in water and sediment of urban lakes in a subtropical city, Guangzhou, South China. AB - Extensive use of phthalate esters (PAEs) in both industrial processes and consumer products has resulted in the ubiquitous presence of these chemicals in the environment. This study reports the first data on the concentrations of 16 phthalate esters (PAEs) in water and sediments of the urban lakes in Guangzhou City. PAEs were detected in all samples analyzed, mainly originating from urban stormwater runoff, atmospheric deposition, as well as untreated discharge of industrial wastewater and municipal sewage. The Sigma(16)PAEs concentrations in water and sediments ranged from 1.69 to 4.72 microg L(-1) and 2.27 to 74.94 microg g(-1)-dry weight (dw), with the mean concentrations of 2.91 microg L(-1) and 20.85 microg g(-1)-dw, respectively, which indicates that sediment is a significant sink for PAEs. Variability of the Sigma 16PAEs concentrations in water and sediment in the urban lakes was almost consistent. The spatial distribution of PAEs was site-specific. Of the 16 PAEs, DMP, DEP, DnBP, DiBP, DMPP, and DEHP were present in all water and sediment samples. DnBP was abundant in water (53.0-81.2%), while no single dominant congener was found in sediments. The abundances of DiBP were similar to those of DEHP, and DiBP and DEHP collectively accounted for 77.2-97.6% of the Sigma 16PAEs concentrations. Congener specific analysis confirmed that DnBP was a predictive indicator for the dissolved summation operator16 PAEs concentration (correlation coefficient r=0.968, p<0.01), and that DiBP was a predictive indicator for the sediment summation operator16 PAEs concentration (r=0.975, p<0.01). As compared to the results for other studies, the urban lakes of Guangzhou were moderately polluted by PAEs. PMID- 17915328 TI - Tycho Brahe: emperor of Hven and the heavens. AB - Although the Danish astronomer Tycho Brahe reacted against Copernicus by restoring the earth to the centre of the cosmos, he built sophisticated instruments to compile massive sets of accurate data. A skilled self-publicist, he commissioned portraits of himself as he established a new type of astronomical research. PMID- 17915329 TI - Hypothesis: TCR signal transduction--A novel tri-modular signaling system. AB - Antigenic peptides initiate an immune response in T cells via the T cell receptor (TCR). The TCR itself is widely regarded as one of the most complex receptors in nature, as it is comprised of at least six different subunits, the antigen recognizing TCRalpha and beta chains, and the signal transmitting CD3deltavarepsilon, gammaepsilon, and zeta2 dimers. In order for a signal to be transmitted from the TCR to the cytoplasm, the CD3 chains must "sense" that an antigenic peptide has been presented to the TCRalpha and beta subunits. After accomplishing this, there are a total of 10 different immunoreceptor tyrosine activation motifs (ITAMs) present within the CD3 chains which effectively activate the T cell and hence the immune response. The importance of each CD3 chain and subsequently each ITAM has been the focus of intense research. However, the precise role(s) played by each CD3 chain has remained elusive. Using the immunomodulatory peptide termed core peptide (CP), which is proposed to inhibit TCR activation by disrupting TCR-CD3 interactions, a tri-modular signaling system for T cell activation is proposed. By contrast to the existing two distinct signaling model (zeta2, CD3epsilongamma/epsilondelta), in this model each of the three dimers, CD3gammaepsilon, deltaepsilon, and zeta2, are proposed to act as three separate and distinct signaling modules, performing both specific and redundant functions. PMID- 17915331 TI - L-arginine transporters in cardiovascular disease: a novel therapeutic target. AB - The amino acid l-arginine participates in a variety of key biochemical and physiological activities, including its well-recognized role as the key substrate for nitric oxide (NO) biosynthesis. The current review describes the cellular influences on arginine metabolism with particular focus on the transport of l arginine in the endothelium. It details the processes by which intracellular and extracellular levels of l-arginine may influence nitric oxide production and further documents the imbalance that is evident in various cardiovascular disease states. In man, impairment of l-arginine transport has been observed in hypertension, heart failure, and renal disease, and it may thus be a relevant therapeutic target for rectification of nitric oxide pathogenesis in these conditions. PMID- 17915330 TI - Immune evasion by acquisition of complement inhibitors: the mould Aspergillus binds both factor H and C4b binding protein. AB - Pathogenic fungi represent a major threat particularly to immunocompromised hosts, leading to severe, and often lethal, systemic opportunistic infections. Although the impaired immune status of the host is clearly the most important factor leading to disease, virulence factors of the fungus also play a role. Factor H (FH) and its splice product FHL-1 represent the major fluid phase inhibitors of the alternative pathway of complement, whereas C4b-binding protein (C4bp) is the main fluid phase inhibitor of the classical and lectin pathways. Both proteins can bind to the surface of various human pathogens conveying resistance to complement destruction and thus contribute to their pathogenic potential. We have recently shown that Candida albicans evades complement by binding both Factor H and C4bp. Here we show that moulds such as Aspergillus spp. bind Factor H, the splicing variant FHL-1 and also C4bp. Immunofluorescence and flow cytometry studies show that the binding of Factor H and C4bp to Aspergillus spp. appears to be even stronger than to Candida spp. and that different, albeit possibly nearby, binding moieties mediate this surface attachment. PMID- 17915332 TI - The nerve growth factor and its receptors in airway inflammatory diseases. AB - The nerve growth factor (NGF) belongs to the neurotrophin family and induces its effects through activation of 2 distinct receptor types: the tropomyosin-related kinase A (TrkA) receptor, carrying an intrinsic tyrosine kinase activity in its intracellular domain, and the receptor p75 for neurotrophins (p75NTR), belonging to the death receptor family. Through activation of its TrkA receptor, NGF activates signalling pathways, including phospholipase Cgamma (PLCgamma), phosphatidyl-inositol 3-kinase (PI3K), the small G protein Ras, and mitogen activated protein kinases (MAPK). Through its p75NTR receptor, NGF activates proapoptotic signalling pathways including the MAPK c-Jun N-terminal kinase (JNK), ceramides, and the small G protein Rac, but also activates pathways promoting cell survival through the transcription factor nuclear factor-kappaB (NF-kappaB). NGF was first described by Rita Levi-Montalcini and collaborators as an important factor involved in nerve differentiation and survival. Another role for NGF has since been established in inflammation, in particular of the airways, with increased NGF levels in chronic inflammatory diseases. In this review, we will first describe NGF structure and synthesis and NGF receptors and their signalling pathways. We will then provide information about NGF in the airways, describing its expression and regulation, as well as pointing out its potential role in inflammation, hyperresponsiveness, and remodelling process observed in airway inflammatory diseases, in particular in asthma. PMID- 17915333 TI - Protease-activated receptors in the brain: receptor expression, activation, and functions in neurodegeneration and neuroprotection. AB - Protease-activated receptors (PARs) are G protein-coupled receptors that regulate the cellular response to extracellular serine proteases, like thrombin, trypsin, and tryptase. The PAR family consists of four members: PAR-1, -3, and -4 as thrombin receptors and PAR-2 as the trypsin/tryptase receptor, which are abundantly expressed in the brain throughout development. Recent evidence has supported the direct involvement of PARs in brain development and function. The expression of PARs in the brain is differentially upregulated or downregulated under pathological conditions in neurodegenerative disorders, like Parkinson's disease, Alzheimer's disease, multiple sclerosis, stroke, and human immunodeficiency virus-associated dementia. Activation of PARs mediates cell death or cell survival in the brain, depending on the amplitude and the duration of agonist stimulation. Interference or potentiation of PAR activation is beneficial in animal models of neurodegenerative diseases. Therefore, PARs mediate either neurodegeneration or neuroprotection in neurodegenerative diseases and represent attractive therapeutic targets for treatment of brain injuries. Here, we review the abnormal expression of PARs in the brain under pathological conditions, the functions of PARs in neurodegenerative disorders, and the molecular mechanisms involved. PMID- 17915334 TI - Neurobiology of orofacial proprioception. AB - Primary sensory fibers innervating the head region derive from neurons of both the trigeminal ganglion (TG) and mesencephalic trigeminal nucleus (MTN). The trigeminal primary proprioceptors have their cell bodies in the MTN. Unlike the TG cells, MTN neuronal somata are centrally located within the brainstem and receive synaptic inputs that potentially modify their output. They are a crucial component of the neural circuitry responsible for the generation and control of oromotor activities. Gaining an insight into the chemical neuroanatomy of the MTN is, therefore, of fundamental importance for the understanding of neurobiology of the head proprioceptive system. This paper summarizes the recent advances in our knowledge of pre- and postsynaptic mechanisms related to orofacial proprioceptive signaling in mammals. It first briefly describes the neuroanatomy of the MTN, which is involved in the processing of proprioceptive information from the face and oral cavity, and then focuses on its neurochemistry. In order to solve the puzzle of the chemical coding of the mammalian MTN, we review the expression of classical neurotransmitters and their receptors in mesencephalic trigeminal neurons. Furthermore, we discuss the relationship of neuropeptides and their corresponding receptors in relaying of masticatory proprioception and also refer to the interactions with other atypical neuromessengers and neurotrophic factors. In extension of previous inferences, we provide conclusive evidence that the levels of transmitters vary according to the environmental conditions thus implying the neuroplasticity of mesencephalic trigeminal neurons. Finally, we have also tried to give an integrated functional account of the MTN neurochemical profiles. PMID- 17915336 TI - Capacitive biosensor for detection of endotoxin. AB - A capacitive biosensor for the detection of bacterial endotoxin has been developed. Endotoxin-neutralizing protein derived from American horseshoe crab was immobilized to a self-assembled thiol layer on a biosensor transducer (Au). Upon injection of a sample containing endotoxin, a decrease in the observed capacitive signal was registered. Endotoxin could be determined under optimum conditions with a detection limit of 1.0 x 10(-13) M and linearity ranging from 1.0 x 10(-13) to 1.0 x 10(-10) M. Good agreement was achieved when applying endotoxin preparations purified from an Escherichia coli cultivation to the capacitive biosensor system, utilizing the conventional method for quantitative endotoxin determination, the Limulus amebocyte lysate test as a reference. The capacitive biosensor method was statistically tested with the Wilcoxon signed rank test, which proved the system is acceptable for the quantitative analysis of bacterial endotoxin (P<0.05). PMID- 17915337 TI - Abstracts of the 2nd BBBB Conference on Pharmaceutical Sciences, September 13-15, 2007, Tallinn-Tartu, Estonia. PMID- 17915338 TI - Abstracts of the 55th International Congress and Annual Meeting of the Society for Medicinal Plant Research, September 2-6, 2007, Graz, Austria. PMID- 17915339 TI - Abstracts of the 7th World Congress on Trauma, Shock, Inflammation and Sepsis (TSIS), March 13-17, 2007, Munich, Germany. PMID- 17915335 TI - A role for the androgen receptor in the sexual differentiation of the olfactory system in mice. AB - Olfactory signals play a central role in the identification of a mating partner in rodents, and the behavioral response to these cues varies markedly between the sexes. As several other sexually dimorphic traits, this response is thought to differentiate as a result of exposure of the developing individual to gonadal steroids, but both the identity of the specific steroid signal and the neural structures targeted for differentiation on this particular case are largely unknown. The present review summarizes results obtained in our lab using genetic males affected by the testicular feminization syndrome (Tfm) as experimental model, and that led to the identification of a role for non-aromatized gonadal steroids acting through the androgen receptor (AR) in the differentiation of olfactory cues processing in mice. The existing literature about AR-mediated sexual differentiation of the CNS in animal models is discussed, along with potential targets for the action of non-aromatized gonadal steroids in either one of the subsystems that detect and process olfactory information in rodents. PMID- 17915340 TI - Abstracts of the 48th International Conference on the Bioscience of Lipids, September 4-8, 2007, Turku, Finland. PMID- 17915341 TI - Abstracts of the 5th International Symposium on Targeted Anticancer Therapies, 8 10 March 2007, Amsterdam, The Netherlands. PMID- 17915342 TI - Measuring the quality of dying in long-term care. AB - OBJECTIVES: To describe two versions of a new measure, The Quality of Dying in Long-Term Care, for postdeath administration to surrogate respondents (staff and family caregivers) of all decedents (QOD-LTC) and of cognitively intact decedents (QOD-LTC-C) who die in nursing homes (NHs) and residential care or assisted living (RC/AL) facilities. DESIGN: Using two levels of exploratory factor analysis, 15 candidate items for the QOD-LTC and 36 candidate items for the QOD LTC-C were tested using multiple criteria to determine factor structure and interpretability of the quality of dying in long-term care (LTC). SETTING: One hundred seventeen RC/AL facilities and 31 NHs in FL, MD, NC, and NJ. PARTICIPANTS: Family (n=439) and staff (n=332) caregivers of 633 decedents from a stratified random sample from RC/AL facilities and NHs in four states. MEASUREMENTS: Trained interviewers asked respondents to rate statements describing potentially important aspects of the quality of dying using a 5-point Likert scale. The scale items were analyzed using exploratory factor analysis with communalities set to unity using a Promax rotation. RESULTS: The 11-item QOD LTC (alpha=0.66), appropriate for surrogate respondents for all decedents, consists of three domains (personhood, closure, preparatory tasks). The 23-item instrument for surrogate respondents of cognitively intact decedents, the QOD-LTC C (alpha=0.85), consists of five domains (sense of purpose, closure, control, social connection, preparatory tasks). CONCLUSION: The QOD-LTC and QOD-LTC-C are psychometrically acceptable measures of the quality of the dying experience, developed for and tested in LTC settings. Use of these measures can increase understanding of the dying experience in LTC. PMID- 17915343 TI - Symptoms of restless legs syndrome in older adults: outcomes on sleep quality, sleepiness, fatigue, depression, and quality of life. AB - OBJECTIVES: To compare differences in sleep quality, sleepiness, fatigue, depression, and quality of life according to severity of symptoms of restless legs syndrome (RLS) in older adults. DESIGN: Descriptive, comparative study; cross-sectional design. SETTING: Penn Sleep Center at the University of Pennsylvania and RLS support groups in Philadelphia. PARTICIPANTS: Thirty-nine adults, aged 65 and older, diagnosed with RLS with symptoms at least 3 nights per week. Participants were stratified according to symptom severity based on scores from the RLS Symptom Severity Scale. Exclusion criteria were dementia, cognitive impairments, and sleep disorders other than RLS. MEASUREMENTS: Sleep quality, measured using the Pittsburgh Sleep Quality Index (PSQI), was the primary outcome. Secondary outcomes were sleepiness, fatigue, depression, and quality of life measured using the Epworth Sleepiness Scale (ESS), Fatigue Severity Scale (FSS), Center for Epidemiological Studies--Depression Scale (CES-D), and RLS Quality of Life Instrument (RLS-QLI), respectively. RESULTS: Significant differences were found in subjective sleep quality (P=.007) and sleep duration (P=.04), as well as in PSQI global score (P=.007). RLS-QLI sleep quality (beta= 0.12, 95% confidence interval (CI)=-0.18 to -0.06, P<.001) and sleepiness (beta=0.35, 95% CI=0.09-0.61, P=.01) were significantly related to PSQI global score. Subjects with severe symptoms were five times as likely to use medication to treat RLS (OR=5.3, 95% CI=1.2-22.2). CONCLUSION: The severity of RLS symptoms in older adults affects not only sleep quality but also many aspects of quality of life, including social functioning, daily functioning, and emotional well being. PMID- 17915344 TI - Computerized physician order entry with clinical decision support in long-term care facilities: costs and benefits to stakeholders. AB - Nursing homes are the setting of care for growing numbers of our nation's older people, and adverse drug events are an increasingly recognized safety and quality concern in this population. Health information technology, including computerized physician/provider order entry (CPOE) with clinical decision support (CDS), has been proposed as an important systems-based approach for reducing medication errors and preventable drug-related injuries. This article describes the costs and benefits of CPOE with CDS for the various stakeholders involved in long-term care (LTC), including nurses, physicians, the pharmacy, the laboratory, the payer (e.g., the insurer), nursing home residents, and the LTC facility. Critical barriers to adoption of these systems are discussed, primarily from an economic perspective. The analysis suggests that multiple stakeholders will incur the costs related to implementation of CPOE with CDS in the LTC setting, but the costs incurred by each may not be aligned with the benefits, which may present a major barrier to broad adoption. Physicians and LTC facilities are likely to bear a large burden of the costs, whereas residents and payers will enjoy a large portion of the benefits. Consideration of these costs and benefits suggests that financial incentives to physicians and facilities may be necessary to encourage and accelerate widespread use of these systems in the LTC setting. PMID- 17915346 TI - American Society for Mass Spectrometry directory of members, 2007. PMID- 17915347 TI - Abstracts of the 30th Annual Meeting of the Japan Neuroscience Society, September 10-12, 2007, Yokohama, Japan. PMID- 17915345 TI - An outreach program improved osteoporosis management after a fracture. AB - This longitudinal retrospective cohort study evaluated implementation of an intervention to improve management of osteoporosis after a fracture in a nonprofit group-model health maintenance organization (HMO) in the U.S. Pacific Northwest with 480,000 members and electronic medical record data. Participants were female HMO members aged 67 and older who sustained a qualifying clinical fracture(s) and who had not received a bone mineral density (BMD) measurement or osteoporosis treatment in the 12 months before the fracture (N=3,588). Phase 1 included outreach to clinicians and patients; Phase 2 added clinician and staff incentives. Primary outcome was "osteoporosis management"--receipt of a BMD measurement or osteoporosis medication in the 6 months after an index fracture. Before the intervention, 13.4% (95% confidence interval (CI)=12.0-14.8%) of patients had received osteoporosis management, and the time trend was not significant. Post-intervention, the probability of osteoporosis management increased on average 3.1% (95% CI=2.6-3.5%) every 2 months throughout both study phases without a significant added improvement in Phase 2. Improvement varied according to clinic and was less likely for patients with dementia. Overall, the probability of osteoporosis management increased from the baseline level of 13.4% to 44.0% (95% CI=40.0-48.0%) by the end of the study period (20 months post intervention). The study found that an outreach program to primary care providers and patients improved the management of osteoporosis after a fracture. If widely implemented, this intervention could substantially improve the secondary prevention of osteoporosis. More-individualized interventions may be necessary for high-risk subgroups. PMID- 17915348 TI - Abstracts of the 5th Annual British Thoracic Oncology Group (BTOG) Meeting, 24-25 January 2007, Dublin, Ireland. PMID- 17915349 TI - Abstracts of the 36th Annual Scientific Meeting of the ISEH (Society for Hematology and Stem Cells), September 28-30, 2007, Hamburg, Germany. PMID- 17915350 TI - Solution structure of a small protein containing a fluorinated side chain in the core. PMID- 17915351 TI - [Picture of the month: vena cava superior syndrome]. PMID- 17915353 TI - Expression profiling and disseminated intravascular coagulation: finding genes gone wild. PMID- 17915352 TI - [Theme issue about sex differences]. PMID- 17915354 TI - [Small hospitals--dangerous romanticization]. PMID- 17915355 TI - Natural orifice transluminal endoscopic surgery. PMID- 17915356 TI - Getting dumped. PMID- 17915357 TI - Does progressive stage transition mean getting better? A test of the Transtheoretical Model in alcoholism recovery. AB - AIMS: To test two central assumptions of stage movement in the Transtheoretical Model (TTM) vis-a-vis alcoholism recovery: (assumption 1) individuals making a forward transition to the action-oriented stages (i.e. preparation/action) will manifest relatively greater drinking improvements than their counterparts remaining in the pre-action stages (i.e. pre-contemplation, contemplation); and (assumption 2) individuals remaining in the pre-action stages across time will not demonstrate clinically relevant improvement in drinking outcomes. DESIGN AND SETTING: Secondary data analyses of data from Project MATCH, a large multi-site alcoholism treatment-matching study. MEASUREMENTS: At baseline and 3 months post treatment, the following variables were measured: stage-of-change (based on the University of Rhode Island Change Assessment measure and the most recent stage assignment algorithm), drinks per drinking day (DDD) and percentage days abstinent (PDA). FINDINGS: Six of the eight tests of assumptions 1 and 2 failed to support the basic tenets of the TTM. Our study demonstrated that individuals making a progressive stage transition to the action-oriented stages (i.e. preparation/action) do not necessarily manifest greater improvements in drinking related behavior than individuals remaining in the pre-action stages (i.e. pre contemplation, contemplation), and that individuals remaining in the pre-action stages over time actually do manifest statistically significant and clinically important improvements in drinking-related behavior. CONCLUSIONS: Our findings challenge not only the criterion validity associated with stage movement in the TTM account of alcoholism recovery, but also recent TTM-based substance abuse treatment approaches which systematically promote forward stage transition as a primary clinical goal and marker of therapeutic success. PMID- 17915358 TI - Do reefs drive diversification in marine teleosts? Evidence from the pufferfishes and their allies (Order Tetraodontiformes). AB - A major challenge in evolutionary biology lies in explaining patterns of high species numbers found in biodiversity hot spots. Tropical coral reefs underlie most marine hot spots and reef-associated fish faunas represent some of the most diverse assemblages of vertebrates on the planet. Although the standing diversity of modern reef fish clades is usually attributed to their ecological association with corals, untangling temporal patterns of codiversification has traditionally proved difficult. In addition, owing to uncertainty in higher-level relationships among acanthomorph fish, there have been few opportunities to test the assumption that reef-association itself leads to higher rates of diversification compared to other habitats. Here we use relaxed-clock methods in conjunction with statistical measures of species accumulation and phylogenetic comparative methods to clarify the temporal pattern of diversification in reef and nonreef-associated lineages of tetraodontiforms, a morphologically diverse order of teleost fish. We incorporate 11 fossil calibrations distributed across the tetraodontiform tree to infer divergence times and compare results from models of autocorrelated and uncorrelated evolutionary rates. All major tetraodontiform reef crown groups have significantly higher rates of diversification than the order as a whole. None of the nonreef-associated families show this pattern with the exception of the aracanid boxfish. Independent contrasts analysis also reveals a significantly positive relationship between diversification rate and proportion of reef associated species within each family when aracanids are excluded. Reef association appears to have increased diversification rate within tetraodontiforms. We suggest that both intrinsic factors of reef habitat and extrinsic factors relating to the provincialization and regionalization of the marine biota during the Miocene (about 23-5 MY) played a role in shaping these patterns of diversity. PMID- 17915359 TI - Daniel E. Koshland, Jr. 1902-2007. PMID- 17915360 TI - Study links viagra to infertility. PMID- 17915361 TI - OB care in family medicine. PMID- 17915362 TI - A revolution in the making. PMID- 17915363 TI - A revolution in the making. PMID- 17915365 TI - [Rethinking in pain therapy. Thus you grab backache in its roots]. PMID- 17915364 TI - Clarification. "Billing for Medicare Part D vaccines.". PMID- 17915366 TI - [Mild cognition disorders signal increased risk of dementia. How can mental decline be stopped?]. PMID- 17915367 TI - [They should make thermotherapy more effective. Hot nanoparticles against cancer]. PMID- 17915368 TI - [Don't wait until the stream dries up--recommend BPH screening]. PMID- 17915369 TI - [Observational study of an AT1-antagonist]. PMID- 17915370 TI - [Intestinal cancer precautions. Stool test for tumor M2 pyruvate kinase]. PMID- 17915371 TI - [Myocardial infraction--genetic detection]. PMID- 17915372 TI - Herd immunity. PMID- 17915373 TI - Multidisciplinary care plans. PMID- 17915374 TI - Bevacizumab on the PBS. PMID- 17915375 TI - Children and autism--Part 1--recognition and pharmacological management. AB - BACKGROUND: Autism is a neurodevelopmental disorder characterized by complex aetiology, variable presentation and widely divergent outcomes. It is clear that an individual's intrinsic genetic susceptibility, health, nutritional status and environmental exposures all contribute to the aetiology of autism. OBJECTIVE: This article aims to assist the general practitioner in recognizing and managing a child with an autistic disorder. DISCUSSION: Screening for autism by the GP can lead to referral for a formal diagnosis, enabling much needed support at an early stage of development, which can improve outcomes for the individual. Currently, evidence for psychotropic use and awareness of adverse effects in young people with autism is limited. Antipsychotic medications are increasingly used in people with autism and the importance of monitoring for adverse effects is paramount. Primary strategies for dealing with children with autism are understanding, environmental modification and behavioural interventions. Combined with these, pharmacological interventions may have benefits for children with autism with extreme or challenging behaviours. PMID- 17915376 TI - Enzastaurin, a targeted PKCBeta inhibitor, in relapsed or refractory DLBCL: a promising new strategy based on gene expression signature. PMID- 17915377 TI - Autologous transplantation using high-dose radioimmunotherapy conditioning in patients 60 years and older. PMID- 17915378 TI - Sentinel-node biopsy or nodal observation in melanoma. PMID- 17915379 TI - Acute fasciolosis diagnosed in young Scottish calves. PMID- 17915380 TI - [Efficacy of sinuforte monotheraphy in patients with acute and chronic rhinosinusitis at an exacerbation stage]. PMID- 17915381 TI - [A giant lymphoma of the laryngopharynx]. PMID- 17915382 TI - [A case of malignant maxillary osteoblastoclastoma]. PMID- 17915384 TI - [Benign paroxysmal positional vertigo]. PMID- 17915383 TI - [A combined penetrating gunshot wound of the orbit, maxillary sinus and cells of the ethmoid bone]. PMID- 17915385 TI - Operative carpal and hand injuries in children. PMID- 17915386 TI - Re: Amoebiasis: current status in Australia. PMID- 17915387 TI - Rural enrollment in Medicare Advantage growing rapidly in 2007, especially in private fee-for-service plans. AB - Enrollment in Medicare Advantage (MA) plans has more than tripled since the inception of the MA program at the beginning of 2006. However, rural enrollment remains well below urban enrollment as a percentage of the eligible population. This policy brief provides findings about enrollment in the newly designed MA program in rural and urban areas across the United States and updates early findings from analysis of the Medicare+Choice/ MA program presented in previous RUPRI Center policy briefs. KEY FINDINGS: As of June 5, 2007 (date of release by CMS), (1) Over 780,000 rural Medicare beneficiaries were enrolled in an MA plan, an increase of 50% since November 2006, and a 222% increase since 2005. (2) Despite significant growth in MA plans, only 8.6% of rural persons were enrolled in MA plans in June 2007, compared to 21.7% of urban persons. (3) Over half (55%) of rural persons enrolled in MA or prepaid plans were in private fee-for-service (PFFS) plans, compared to only 14% of urban persons. (4) PFFS enrollment in rural areas in June 2007 was concentrated in several PFFS plans, with almost 90% of rural persons enrolled in plans run by seven organizations serving about 2,000 counties in the United States. PMID- 17915389 TI - The risk of risk reduction: can postmarket surveillance pose more risk than benefit? PMID- 17915388 TI - Cause and effect? Assessing postmarketing safety studies as evidence of causation in products liability cases. PMID- 17915390 TI - Resistance to arranged marriage among Nubian youth: ideology and changing times. PMID- 17915391 TI - Pharmaceutical postmarket review: fact or fiction? PMID- 17915392 TI - Drug safety: a contrarian's point of view. PMID- 17915393 TI - U.S. postmarketing pharmacovigilance compliance in the midst of regulatory uncertainty. PMID- 17915394 TI - Testimonies in exile: Sudanese gender politics. PMID- 17915396 TI - The drug safety system conundrum. PMID- 17915395 TI - Red sky in the morning: modifying prescription drug labels as a result of postmarket surveillance. PMID- 17915397 TI - Pin the tail on the other donkey: allocating and avoiding injury losses after drug or device approval. PMID- 17915398 TI - Medical device reporting: issues with Class III medical devices. PMID- 17915399 TI - Marriage, Sudanese-style: transnational practices of citizenship and gender making for Sudanese nationals in Egypt. PMID- 17915400 TI - Medical device recalls from 2004 to 2006: a focus on Class I recalls. PMID- 17915401 TI - Medical device safety: FDA's postmarket transformation initiative. PMID- 17915402 TI - Medical devices post-approval studies program: vision, strategies, challenges and opportunities. PMID- 17915403 TI - Strategy for surveillance of adverse drug events. PMID- 17915404 TI - Claiming the campus for female students in Bangladesh. PMID- 17915405 TI - Online digital imaging academy. PMID- 17915406 TI - Oral versus non-oral hormone replacement therapy: how important is the route of administration? AB - Although there are differences in the pharmacokinetic profiles of oral and non oral routes of administration the clinical relevance of these differences remains to be determined. Likewise, there are differences in the metabolic and haemostatic effects of different routes of administration of oestrogen but these may have clinical relevance. For some parameters, such as lipids and lipoproteins, glucose and insulin metabolism, there are greater benefits from oral administration; for others, particularly haemostatic changes and effects on CRP, there are advantages from transdermal administration. For the potential benefits of HRT on CHD, these differences probably have less impact than the effect of the dose of hormones used and the lowest effective should be prescribed. Irrespective of dose, certain small sub-groups of patients should be specifically treated with an oral regimen eg those with lipid and lipoprotein abnormalities and impaired glucose tolerance whereas others should be treated with a transdermal regimen eg those with a personal or relevant family history of venous thrombosis. However, the vast majority of patients possess none of these risk factors and for them it will come down to personal preference. The availability of different combinations and doses of hormones, as well as different routes of administration, allows HRT to be tailored to the individual and there are few women for whom a suitable form of HRT cannot be found. Although data are lacking we believe it unwise to believe that fully transdermal combination therapy will not impact on risk of incident breast cancer. Based on current evidence transdermal HRT may also cause more irregular and breakthrough bleeding with sequential and continuous therapies than oral counterparts. PMID- 17915407 TI - The evaluation of selected indices of apoptosis in placentas from pregnancies complicated by fetal growth restriction. AB - BACKGROUND: Fetal growth restriction is related to a high rate of prematurity and mortality. In cases of unknown origin utero-placental circulation changes are the main factor which is due to the changes in blood vessels. The understanding of the mechanism may help in further prevention of FGR. MATERIAL AND METHODS: The expression of bcl-2 and bax in normal pregnancies and complicated by FGR were compared. The study was conducted in 2005-2006 at The Medical University of Lodz- HRP Unit and The Kopernik Hospital--Lodz. Bcl-2 and bax were estimated using an immunohistochemical method. Bcl-2 was estimated in trophoblast, bax in decidua and trophoblast. RESULTS: In a study group the mean value of bcl-2 in trophoblast was 37.04 +/- 10.51, in a control group the mean value was 65.74 +/- 6.97. The estimation of bax was done in trophoblast and decidua separately. In the group of FGR mean value of bax expression in trophoblast was 45.35 +/- 10.5. In decidua the mean bax expression value was 24.11 +/- 7.3. In controls in trophoblast the mean value was 12.53 +/- 7.54, in decidua the mean expression of bax was 6.63 +/- 2.24. CONCLUSION: 1. Apoptosis in trophoblast is lower in normal pregnancy than in FGR. 2. Increased expression of pro-apoptotic proteins in placenta might be one of the reason for FGR development. PMID- 17915409 TI - [Anxiety assessment in parturients requesting epidural analgesia for pain relief]. AB - OBJECTIVES: Neuraxial methods provide the most effective labor pain relief. This study aimed at assessing anxiety level in parturients requesting epidural analgesia (EA). MATERIAL AND METHODS: Forty five women in spontaneous, active labor were enrolled, both primiparas (n=36) and multiparas (n=9). Anxiety was assessed by means of Spielberger State and Trait Anxiety Inventory (STAI) before administration of EA, and pain was measured by visual-analog scale (VAS) before and after analgesia. RESULTS: In all the studied parturients state anxiety was strikingly higher than the trait (53.9 +/- 11.8 vs. 39.3 +/- 8.4; P < 0.0001); the difference appeared insignificant in multiparas only. State anxiety was comparable independently of parity, labor outcome and systemic opioid administration. No association between anxiety level and labor pain intensity preceding analgesia, the duration of labor stages and demographic parameters could be found. However, a negative correlation between state anxiety and pain intensity reported after EA administration was noted (R = -0.315, p = 0.040), and, in cases of physiological labor, a negative association between state anxiety and the neonate Apgar score at the 1st minute after birth could be observed (R = -0.337, p = 0.047, Spearman rank test). CONCLUSIONS: In parturients requesting EA, state anxiety level is increased and not connected with the trait. Furthermore, in these women, anxiety appears not to be associated with labor pain but may influence the analgesic effect of the blockade. Anxiety does not determine labor duration and outcome; however, it may be connected with the well being of the neonate immediately after birth. PMID- 17915408 TI - [Heterogeneity of insulin resistance level in gestational diabetes mellitus. Therapeutic implications]. AB - OBJECTIVE: The evaluation of insulin resistance (IR) level in population of women with gestational diabetes(GDM) and its relation to treatment of GDM. MATERIALS AND METHODS: 657 GDM women, aged 17-45, treated between the years 2003 and 2005, in Bydgoszcz were studied. Age, pregravid body mass index(BMI), weight gain during pregnancy at the GDM diagnosis, week of GDM diagnosis, week of the beginning of insulin therapy and daily doses of insulin were assessed in the whole population. Daily doses of insulin were evaluated as minimal doses needed at the initial phase of GDM therapy and as maximal doses during gestation. IR was evaluated at the GDM diagnosis, with the use of homeostasis model assessment (HOMA-IR), based on fasting glucose and insulin concentration. RESULTS: 47% women were classified as low HOMA-IR(<2) subpopulation, 50% as intermediate HOMA-IR(2 10) subpopulation, 3% as high HOMA-IR(10-46)subpopulation. Subpopulation with intermediate HOMA-IR had higher BMI, higher weight gain and blood glucose at 0 OGTT compared to subpopulation with low HOMA-IR but lower insulin concentration compared to high HOMA-IR subpopulation. Women in high HOMA-IR subpopulation and in intermediate HOMA-IR subpopulation were twice as often treated with insulin, compared to low HOMA-IR group, accordingly, 58%, 42%, 24%. Daily insulin doses, assessed both minimal and maximal doses, were increasing parallel to HOMA-IR in whole population, accordingly, minimal doses of insulin, 16.0 = -12.7 vs 18.4 vs 20.8 vs 30.8 +/- 30.3 and maximal doses of insulin, accordingly, 39.0 +/- 322.4 vs 50.9 +/- 42.4 vs 70.3 +/- 30.3. CONCLUSION: The studied population of women consisted mainly of subpopulation with low or intermediate HOMA-IR value, in rare cases, of high HOMA-IR value. Our results suggest that adipose tissue is particularly associated with insulin resistance level in subpopulation with intermediate HOMA-IR. Both, frequency of insulin therapy and daily insulin doses are associated with insulin resistance level at the GDM diagnosis. PMID- 17915410 TI - [The insertion/deletion polymorphism (I/D) of the ACE gene in pregnant women with excessive weight gain]. AB - INTRODUCTION: The body mass gain is conditioned by lifestyle, as well as many environmental and genetic factors. Recent studies suggest that renin-angiotensin system (RAS) plays a fundamental role in process of growth and differentiation of adipocytes through the acting of angiotensin II and seems to be a significant factor in excessive weight gain development. The purpose of this study was to determine the frequency and significance of insertion/deletion polymorphism (I/D) of the ACE gene in pregnant women with excessive weight gain. MATERIALS AND METHODS: The examined group consisted of 212 pregnant women, including 107 women with normal (DeltaBMI< or =5) and 105 women with excessive weight gain (DeltaBMI>5). Genomic DNA was extracted from venous blood. The I/D polymorphism of ACE gene was determined by polymerase chain reaction (PCR). RESULTS: During the course of the study we did not observe the statistically significant higher frequency of ACE genotypes in any of the two investigated groups of women with normal and excessive weight gain. Nevertheless, an overrepresentation of II genotype frequency in group with excessive weight gain has been observed (33.3 vs 21.5%, p=ns). The same findings were visible as far as the frequency of I allele in group with excessive weight gain was concerned (55.2 vs 45.8%, p=ns). The frequency of observed genotypes was in agreement with Hardy-Weinberg equilibrium. CONCLUSIONS: Although overrepresentation of II genotype and I allele in the group of pregnant women with excessive weight gain (DeltaBMI>5) has been observed, a close correlation between II genotype and higher risk of overweight could be not indicated (due to the lack of significant difference). The results should be confirmed in a more numerous group of pregnant women. At this stage the results of the study did not suggested the presence of association of I/D polymorphism of ACE gene with weight gain in investigated group of pregnant women. PMID- 17915411 TI - [Psychological causative factors in postpartum depression amongst women with normal and high-risk pregnancies]. AB - OBJECTIVES: The study presented in the following article was based on the assumption a state of mind of a mother-to-be directly influences her state of mind after giving birth. The aim of the study was to determine whether a case of a high-risk pregnancy, and the related stress, can be significant causative factors in postpartum depression, in case of pregnancies with a normal course, as well as whether incidences of postpartum depression among women with normal and high-risk pregnancies are correlated with the level of stress and depression experienced during pregnancy. MATERIALS AND METHODS: 70 women were examined in the study: 35 women with normal pregnancies represented the control group and 35 women with high-risk pregnancies represented the study group. The following methods were used to find answers to the posed research questions: a demographic survey, Perceived Stress Scale, the Beck Depression Scale and the Edinburgh Postnatal Depression Scale. RESULTS: Women with high-risk pregnancies demonstrated a significantly much higher level of stress and related negative emotions; also they handled the stress and emotions worse than women with normal pregnancies. Also, higher intensity of depression symptoms during pregnancy were observed in the study group of women with high-risk pregnancies. In both groups, higher intensity of symptoms of post-childbirth depression was correlated with higher level of stress and more severe depression during pregnancy. CONCLUSIONS: Pregnancy--especially high-risk pregnancy, which is associated with health difficulties and complications, and consequently, higher stress level, feelings of uncertainty and depression--is an additional risk factor of postpartum depression. There exists a necessity to create therapeutic programs aimed at psychological support for suffering women. PMID- 17915412 TI - [Diagnostic and treatment of overactive bladder]. AB - The overactive bladder (OAB) is characterized by symptoms of frequency, urgency, nocturia, and urge incontinence, substantially affecting the quality of life of millions of people throughout the world. Diagnosis of OAB made on patient history, physical examination, and clinical tests can be used as the basis for treatment in most cases. In cases where there is uncertainty regarding the diagnosis, urodynamic assessment should be carried out. Treatment of OAB includes behavioral therapy, pharmacology and, in some cases, surgery. This article reviews current findings regarding diagnostics and treatment of the overactive bladder. PMID- 17915413 TI - [Diagnostic techniques in breast cancer detection. Part III. The new cytobiological methods]. AB - This publication contains an overview of the cyto-biological technologies utilized in breast cancer diagnosis. It refers to technologies created de novo and heralded as a breakthrough ones, as well as to those existing for years. An integration of new diagnostic biogenetic methods, with imaging techniques such as mammography, have become essential in order to cope with the requirements of early detection of breast tumours. Apart from the new and innovative diagnostic techniques, this publication also presents an outline of recommendations for genetic diagnostics as well as cooperative and multidisciplinary integrative activities, which are prerequisites for successful diagnosis and treatment and, consequently, longer survival periods upon which patients rely. PMID- 17915414 TI - [West Nile Virus infection in perinatology]. AB - West Nile Virus causes one of the so-called emerging infectious diseases and is considered to be one of the major epidemic problems in many countries. Most of the WNV-infected-people do not develop clinical symptoms. Nevertheless, Infection with WNV can lead to severe neurological diseases, classically associated with West Nile fever. Among several neurological manifestations that have been observed, encephalitis is the most typical presentation. Transplacental passage is one of the confirmed transmission ways, but neonatal infection by breast feeding is also probable. Congenital WNV infection has been accompanied by bilateral chorioretinitis and severe malformation of the fetal central nervous system. At present, due to the lack of sufficient data, possibilities to conclude on the nature of the relationship between WNV infection and fetal malformations are limited. PMID- 17915415 TI - [Effective transplacental therapy in fetal cardiomegaly to prevent lung hypoplasia--case report]. AB - In our referral center, 14,481 fetal echocardiography examinations were performed in 10,077 fetuses, in years 1994-2006. Fifty four fetuses presented cardiomegaly HA/CA > 0.6. There was one single survivor whose medical history is presented. Fetal cardiomegaly was detected by obstetrical ultrasound screening. In tertiary center HA/CA was evaluated as the following: 0.5 at 32 wks and 0.65 at 36 wks. In addition to cardiomegaly, pulmonary stenosis/atresia, dysplastic tricuspid valve with massive regurgitation were diagnosed. Digoxin was administered transplacentally for 33 days and, additionally, 30 min. oxygen was provided by mask for pregnant woman, 3 times per day. Spontaneous labour took place at 38 wks, with 3100 birth weight and 9 Apgar score. The newborn baby had only temporary tachypnoe, had a planned postdelivery therapy including prostin and cardiacsurgery and was discharged home at the age of 4 weeks in a good clinical condition. This is the first case in our institution of such massive cardiomegaly who did survived the neonatal period. The possible beneficial effects of transplacental digoxin and oxygen therapy are discussed, as well as fetal echocardiography monitoring using HA/CA and pulmonary venous Doppler flow. PMID- 17915416 TI - Isolated severe hydrothorax with respiratory distress as a main manifestation of ovarian hyperstimulation syndrome preceded by respiratory tract infection caused by Haemophilus influenzae. AB - INTRODUCTION: An isolated unilateral pleural effusion as the only presentation of ovarian hyperstimulation syndrome (OHSS) is very rare. This case is an unusual presentation of OHSS after a confirmed respiratory tract infection, with no other coexisting risk factors identified for this syndrome. We also imply that the presence of Haemophilus influenzae in bronchial fluid can increase local reaction to vasoactive cytokines. CASE REPORT: A 32-year-old woman presented at the Department of Reproductive Medicine and Gynaecology of the Pomeranian Medical University after 10 years of infertility with diagnosed hyperprolactinemia followed by bromocriptine treatment. The patient had three IUIs but no pregnancy was achieved. Therefore, ICSI was proposed. After an ovarian hyperstimulation, oocyte aspiration gave 8 oocytes. Although ICSI was performed in all of the oocytes there were 3 fertilizations. The ET of 3 embryos was carried out following 3 days of culture. Three weeks before the gonadotropin administration and a week before GnRH administration the patient had a respiratory tract infection with the most typical syndromes. The infection was treated successfully with over-the-counter medications and antibiotic. Three days after ET the patient was admitted to the ICU with signs of severe dyspnoea. The chest X-ray showed a large pleural effusion over the right lung. Upon admission, thoracocentesis was preformed and 1600 ml of clear fluid was aspirated. The bronchial aspirate showed evidence of Haemophilus influenzae and leukocytes. After three days of standard treatment the chest X-ray revealed no pathology. The patient was discharged asymptomatic on the 4th day of treatment. Serum beta-hCG level was negative on day 12 after ET. CONCLUSIONS: This case suggest that respiratory tract infection prior to stimulation may constitute a new independent risk factor for OHSS. However, the true relation between the respiratory tract infection and susceptibility to OHSS still awaits explanation. Recent or existing respiratory tract infection may be a relative contraindication for starting COH. PMID- 17915417 TI - [Recommendation of The Polish Gynecological Society: Infection with Chlamydia Trachomatis in obstetrics and gynecology]. PMID- 17915418 TI - [Amniocentesis--early or late during pregnancy?]. PMID- 17915419 TI - [Quo vadis, hypophysis? Some news and prospects]. AB - Knowledge of hypophysis could hardly stagnate in a context of general progress in medical science as such. New knowledge in physiology, pathology and treatment of hypophysis diseases is vast and only some of it could be included in the article. New regulators of hypophysial secretion have been discovered. Among them are hypothalamic chemokines and the KISS-1 gene product--kisspeptin. Impulses coming to the hypophysis from the brain centres and the periphery need to be integrated. This is provided by a system of folliculo-stellate cells, paracrine mechanisms and hypophysial microcirculation. Are there stem cells in the hypophysis, too? It seems there are. It could be the above mentioned folliculo-stellate system cells, or the recently discovered SP (side population) cells. Massive injuries such as multiple traumas, severe burns and shock states provoke a double-phase response of the hypophysis. The acute phase is characterised by hypersecretion of most of hypophysial hormones and peripheral resistance to their actuation. In the subsequent chronic phase, however, the secretion of all hypophysial hormones is reduced, except for ACTH. Clinically relevant hypophysial adenomas affect approximately 1 per thousand of population. Two thirds of the above number are prolactinomas. Most prolactinomas can be cured without major difficulty, only those resistant to pharmacological treatment can become a problem. In such cases, Leksell gamma knife can play an important role. The treatment of acromegaly is far more difficult, though. We have developed our own acromegaly treatment method. We treat adenomas surgically, expose possible residua to Leksell gamma knife irradiation and apply pharmacological therapy until the effect of irradiation has been achieved. The therapeutic options are ordered in a cost sequence: cabergolin, somatostatin analogues, pegvisomant. Similar approach is applied to patients with central etiology Cushing's syndrome, the only difference being the fact that the pharmacological therapy preceding the effect of the gamma knife treatment uses ketoconazol and metyrapone. Like in the case of acromegaly, also in that of Cushing's disease, new drugs are developed which promise greater therapeutic advantages. PMID- 17915420 TI - [Subclinical thyroid diseases]. AB - Subclinical thyroids disease (STD) is recently defined term in clinical thyroidology, which includes mainly functional disorders. Basic diagnostic signs are: normal values of thyroid hormones (fT4, fT3) and elevated TSH level (subclinical hypothyroidism) or suppresed TSH level (subclinical hyperthyroidism). In a category of STD may be included subclinical autoimunne thyroiditis (elevated level of thyroid antigens antibodies and/or hypoechogenity in sonographic screen, increased volume of the thyroid without clinical symptoms and/or autoimminity) and microscopic lesions of papillary thyroid carcinoma. Subclinical hypothyroidism may be dangerous for tendency to development of manifest hypothyroidism and for risk of disorders of lipid profile and development of atherosclerosis and its organ complication (esp. myocardial infarction). Subclinical hyperthyroidism is a risk factor of cardiac arythmias and probably can increase a risk of cardiovascular mortality) as well for osteoporosis (esp. in peri- and post-climacteric women), and last but not least for degenerative diseases of brain (?). Indication of treatment of STD is a matter of controversies. Recomendations of experts, varied from "no therapy, monitoring only" to "treat always". Treatment of risk groups (esp. pregnant women) is probably nowadays a most rationale recommendations since results of sofisticated prospective studies will be available. PMID- 17915421 TI - [Aspiration cytology of the thyroid]. AB - Fine needle aspiration cytology is a basic type of examination in thyreology today. It is primarily indicated for thyroid node to exclude a malignant tumour. The presence of multiple nodes or of one node only has shown to be indicative of the same amount of risk of a malignant tumour for the patient. Ultrasound characteristics are insufficient for the determination of the nature of the focus and all the existing nodes greater than 10 mm are an indication for puncture. As for other diseases, aspiration cytology is indicated in granulomatose thyreoiditis, while in Hashimoto thyreoiditis it is usually indicated only if a node is present. In addition to clearly benignant and clearly malignant results, large part of cytological conclusions fall within the category of suspicious (unclear) findings. All of them are an indication for histological verification, and the predictive value of the results depends on their type. It is approximately 20% in the case of follicular neoplasia, and preoperative histology is useless. Therefore we mostly opt for hemithyreoidectomy, further procedure depending on the final histological finding. PMID- 17915422 TI - [Autoimmune polyglandular syndromes: clinical aspects]. AB - The history of autoimmune polyglandular syndromes (APS) and the basic characteristics of types 1 and 2 are briefly introduced. The clinical aspects of the more common type 2 are discussed in more detail from the point of view of the endocrinological and diabetological practice. The diagnosis aspects of preclinical, subclinical and manifestation stage of the main diseases occurring along with APS are briefly mentioned as well as the treatments and their challenges cause by the combination of various endocrine and non-endocrine autoimmune diseases. It is emphasized that care for there patients is a life-long process and that the health condition of these patients is usually very complicated. In some cases the combination of various disease conditions can make these patients completely disabled. PMID- 17915423 TI - [Endocrinology and nuclear medicine aspects of the treatment of thyroid tumours]. AB - Thyroid tumours belong among the most frequent malignant tumours in endocrinology. The rate of incidence of new carcinomas has increased dramatically in recent years; the rate of thyroid tumour incidence, in particular of its less advanced papillary forms, sextupled in the Czech Republic in the previous decade. The article brings an overview of the factors involved in the development of thyroid tumours, and the classification and clinical assessment of their different types. Recombinant human TSH has started to be used in the Czech Republic as part of preparation of patients for radioiodine treatment. Also discussed is the relevance of tumour markers for the prognosis of the disease in the context of life-long monitoring of patients with thyroid carcinoma. PMID- 17915424 TI - [Hypopituitarism--substitution therapy]. AB - Hypopituitarism is a condition linked with insufficient function of the adenohypophyse and occurs in the case of destruction of its greater part. It may take the form of an isolated deficit or a combined disorder--panhypopituitarism. The most common cause is an expansion in the region ofsella turcica, irradiation, traumas, inflammations, bleeding, puerperal ischemic necrosis (Sheehan syndrome) or an affection of the hypothalamus. The cause remains unclear in the case of idiopathic hypopituitarism. CLINICAL PICTURE: The most common case in children is the deficit of the growth hormone, manifested by retarded growth. Sometimes it is combined with the deficit ofgonadotropins which causes pubertas tarda. The clinical picture in adult patients is proportionate to the degree and type of disorder of the different hypophysial functions. Diagnosing of the disease is based on the clinical picture and laboratory examination of hypophysial hormones together with the respective peripheral gland hormones. Dynamic stimulation tests need to be used for more precise diagnosis in the case of threshold results. THERAPY: Expansive processes in the region of the hypophysis are treated by surgery or irradiation, mostly using the gamma knife. Substitution by peripheral gland hormones is used in persistent hormonal deficits. Hypocorticism is treated by hydrocortizone at a dose of 10-30 mg per day administered per os, respecting the diurnal rhythm. The dose should be adjusted in case of strain, and parenterally administered hydrocortizone is necessary in the case of decompensation, pituitary crisis or surgery. Hypothyreosis is treated by thyroxin, the usual substitution dose ranging between 50 and 150 microg per day. The dose is titrated according to the clinical condition and free thyroxin level. Testosterone substitution therapy is used to treat male hypogonadism. In women, a combination of estrogens and gestagens is used on a cyclic or a continuous basis. If fertility is the therapeutic objective, temporary administration of injection gonadotropins or gonadoliberin is necessary in both the sexes. Therapy with growth hormone is indicated in children and in adults with sever, clinically manifested deficit of GH where its administration clearly improves the quality of life, promotes the growth of lean body mass, an increase in bone density and the improvement of metabolic parametres. PMID- 17915425 TI - [Randomly discovered enlargement in the region of sella turcica]. AB - Computer tomography (CT) and magnetic resonance imaging (MRI) quite often detect unexpected cases of enlargement in the hypothalamus-hypophysial region, without the above methods being indicated for clinical manifested symptomatology provoked by the tumour. This is not surprising if we consider that autopsies show the presence of hypophysial adenomas of 10-15% of population on an average. X ray, CT or MRI are indicated in the case of head traumas, lateral nasal cavity inflammations, headaches, strokes, neurological diseases and other disorders. A number of tumours of diverse etiology occur in the hypothalamus-hypophysial region, but hypophysial adenomas are by far the most frequent among all (above 90 %). Among other primary enlargements, the most frequent are craniopharyngeomas and meningeomas, while other enlargements are by fare less common. Such randomly detected tumours are mostly asymptomatic, but targeted anamnesis may show some of the symptoms quite clearly. The symptomatology can be linked with possible slight hormonal overproduction of hypophysial adenomas, a deficit of hypophysial hormones or local manifestations of expansion. Exact assessment of MRI results, of hormonal activity of the enlargement, of the relation to surrounding structures, especially the optic nerves, and the assessment of hypophysial functions are important for the therapeutic decision. Depending on the type and extension of the tumour the options considered are pharmacotherapy (the treatment of choice in the case of prolactinomas), surgery, radiotherapy (today prevailingly using the gamma knife), and if no intervention is necessary, follow up with regular MRI examinations. Tumorous growth is more often observed in "macroadenomas" than in "microadenomas" (up to 10 mm). PMID- 17915426 TI - [Adrenal incidentalomas]. AB - Adrenal incidentalomas are called as a disease of modern technology because increased availability and accuracy of imaging techniques lead to increased incidence of incidentally discovered adrenal masses. Adrenal incidentalomas are found in about 0.35-4.36% of abdominal CT scans performed for purpose other than suspicion of adrenal diseases. However, in autoptic studies the percentage of incidentally discovered adrenal tumours in much higher, in some of then being even more than 20%. Important component of differential diagnosis is endocrinological examination aimed to reveal potential autonomous hormonal overproduction. Approximately more than 10% of adrenal incidentalomas is characterized by autonomous cortisol production, 5-10% is phaeochromocytomas and 5-7% is aldosteron producing adenomas. Vast majority of adrenal incidentalomas are nonfunctioning tumours, mainly adenomas. Rational ways and recommendations for screening of hormonal activity of adrenal incidentalomas is discussed. Very important issue is to distinguish between benign and malignant lesions and between adenomas and non-adenomas. The accuracy of CT, MR, and fine needle aspiration cytology in distinguishing between adenomas from non-adenomas is comparable, in general not exceeding 90%. Particular characteristics of particular adrenal tumours are discussed. From the relationship between the tumour size and probability of malignancy, current recommendations are derived, that non-functioning tumours smaller than 4 cm are indicated for further follow up, size over 6 cm is indication for adrenalectomy and in tumours between 4-6 cm decision process is complex. PMID- 17915427 TI - [Differential diagnosing of hypercalcemias]. AB - Primary hyperparathyreosis and tumour diseases are the two most frequent causes of hypercalcaemia. Surgical removal of parathyroid adenoma is the permanent solution for hypercalcaemia. Hypercalcaemia may occur in 20-30% of patients with cancer in the course of the disease. It causes progressive deterioration of the overall condition of the patient which culminates in a coma-like state with renal failure and means a bad prognosis for the affected person. Evaluation of clinical condition and obtaining the immunoreactive parathormone level data are of extreme importance for correct diagnosis. Normal or even low parathormone levels almost surely exclude primary hyperparathyreosis as the source of hypercalcaemia. Additional, less frequent causes of hypercalcaemia should also be taken into consideration, such as diseases caused by the granulomatose tissue, familial benign hypocalciuric hypercalcaemia, drug provoked hypercalcaemia, high thyroid hormone doses and patient dehydration. Fast replenishment of liquids and administration of bisphosphonates are the cornerstones of hypercalcaemia therapy. PMID- 17915428 TI - [Glucocorticoids and osteoporosis]. AB - Treatment with glucocorticoids (GC) has no alternative in many medical disciplines for their anti-inflammatory and immunosuppressive effect. However, osteoporosis and the related fractures are a serious complication brought about by long-term GC therapy. The risk of fractures, especially of the vertebras and the ribs, becomes higher as early as in the first months of oral GC therapy. It grows in proportion to the daily dose of GC, and is present even if low doses are administered (2.5-7.5 mg of prednisone per day). Decreasing bone density (BMD) is not accountable for the higher risk of fractures in GC therapy and fractures occur with higher values of BMD than in primary osteoporosis. There is still no tool that we could use to quantify the changes in the bone quality and the increased risk of fracture in clinical practice. The principal mechanism by which GC induces osteoporosis is inhibition of bone formation caused by the suppression of osteoblastogenesis as well as the activity of functional osteoblasts, with accelerated osteocyte and osteoblast apoptosis. There are significant differences between individuals in terms of GC sensitivity, the reasons of which have not yet been explained. Prior to planned long-term GC therapy (> 3 months) with daily doses higher than 2.5 mg of prednisone p.o. (or higher doses of inhaled GC), it is recommended to perform a densitometry exam using dual-energy X-ray absorptiometry (DXA) in the lumbar region of the spine and femoral collum to evaluate additional risk factors of osteoporosis and fractures for a more precise estimate of the risk of fracture in the specific patient. Sufficient intake of calcium (1,000-1,500 mg of elementary calcium per day) and of the vitamin D (800 IU per day) should be assured in all patients treated by GC. Endogenous production of sexagens should be evaluated and possible substitution therapy should be considered in premenopausal women and younger men. Today, bisphosphonates can be given to patients with a high risk of fracture, the effects of which in preventing the decrease of BMD and vertebral fractures have been documented in randomised clinical studies, even though the evaluation of the risk of fractures was not the primary endpoint of those studies. However, in view of the antiremodelling effect of bisphosphonates, it is clear that this therapy does not eliminate the cause of GC induced osteoporosis and drugs with stimulating effect on osteoblasts will certainly be preferred in the future. Very promising are the first clinical studies of injection parathormone (PTH 1-34) which stimulated bone formation in a continuing GC treatment. PMID- 17915429 TI - [Mineral-bone disorder with chronic kidney disease]. AB - Mineral-bone disorder in chronic kidney disease is a clinical syndrome provoked by the combination of three factors: abnormal laboratory results, bone morphology disorder and extra-bone calcification. Its onset in adult age is linked with a decrease in glomerular filtration (GF < 1 ml/s). Fully developed forms occur in the course of regular dialysis treatment. The use of the traditional denomination "renal osteodystrophy" is currently restricted to the bone morphology finding. As there are two threshold types of bone turnover (low and high) and two degrees of mineralisation (low and normal), there is a total of four basic variants of mineral-bone disorder. The high turnover variants--secondary hyperparathyreosis and a combined disorder--are still the most frequent and are diagnosed in 70 to 80% of cases. Low turnover disorders include osteomalatia (OM) and adynamic bone disease (ABD). While OM is becoming increasingly rare, the occurrence of ABD is on the rise. The main reason for this may be the steady growth in the age of dialised patients and a number of risk factors, as well as treatment with inadequately high doses of vitamin D. Progressive chronic kidney disease may be linked with D-hormone deficit, negative calcium balance and with positive phosphate balance. Phosphates become a key factor in the development and progression of secondary hyperparathyreosis and extra-bone calcification in the case of D-hormone substitution. Therefore, maintaining a good phosphate balance by restricting their intake or by reducing their intestinal resorption through the use of phosphate binders is the most efficient therapeutic procedure. In patients with chronic kidney failure, adequate dialysis treatment is necessary. Hyperphosphatemia and extra-bone calcification are new independent risk factors of cardiovascular morbidity and mortality. PMID- 17915430 TI - [Oral treatment of type 2 diabetes mellitus]. AB - The article informs about current views of the ethiopathogenesis of type 2 diabetes mellitus (DM). It presents diabetes as a disease which is chronic and progressive, therefore requiring a dynamic approach to treatment. Based on the above concept, the article lists the current armamentarium of oral antidiabetic drugs and the possibilities of their combining. In addition to the existing therapeutic options, it also brings information about innovative drugs from the above group to be made available in the near future. PMID- 17915431 TI - [Transplantation in the treatment of diabetes]. AB - Diabetes mellitus continues to be the most common cause of chronic kidney failure, blindness acquired in adulthood, non-traumatic amputations and severe forms of neuropathy. Therefore it is necessary to look for new forms of therapy capable of achieving long-term normalisation of blood sugar levels. The only standard method so far is pancreas transplantation. Most often, it is performed in combination with kidney transplantation and only exceptionally as an isolated procedure. A new and considerably less invasive option is transplantation of isolated Langerhans islets. While the number of pancreas transplantations in IKEM has exceeded 300, the program of islet transplantation is in its formative phase, with 10 clinical surgeries having been performed since May 2007. However, the number of suitable patients who could benefit from this method of treatment largely exceeds the availability of organs suitable for transplantation. Therefore, new possibilities of acquiring insulin producing cell lines are searched for, both from animal tissue and, primarily, from embryonic or adult stem cells. Also the possibility of in vivo regeneration of endogenous or transplanted beta cells of the pancreas has now became an object of study. Combined transplantation of the kidney and pancreas is still the best available method in the treatment of uremic type 1 diabetes patients and its long-term results have shown to be very good, even though their further improvement has been but of a lesser degree. Isolated transplantation of the pancreas is still reserved for a limited group of patients with very labile diabetes. The transplantation of isolated Langerhans isletes is an alternative option which is far safer for the patient, but the long-term results of which still leave much to be desired. The method currently used in the Institute of Clinical and Experimental Medicine (IKEM) is organ transplantation of the pancreas while a program of transplantation of isolated islets has been launched, and also studied are the possibilities of insulin producing cell lines propagation. PMID- 17915432 TI - [Recommendations for diagnosis and therapy of chronic heart failure. Czech Cardiology Society 2006]. PMID- 17915433 TI - [Allergies: a new epidemic]. PMID- 17915434 TI - [Epidemiology of allergies in Austria]. PMID- 17915435 TI - Epidemiology of allergies in Austria. Results of the first Austrian allergy report. AB - The first Austrian Allergy Report is a compilation of all available epidemiological data relating to allergies in Austria. According to this report the cumulative prevalence of allergies in the Viennese population is 27.6% for men and 32.2% for women and the period prevalence in the year before questioning 19.6% and 22.4% respectively. 20.8% of men and 23.1% of women reported about allergies at health examinations. However, an allergy sensitivity to at least one inhalation allergen, verifiable by means of a prick test, is detectable in 50.8% of the general population and in 39.3% of those free from ailments. Analysis of the hospital discharge statistics of all Austrian hospitals shows that around 12,000 people per year were admitted on grounds of an allergy. In accordance with international studies some population sub groups are more often affected than others. Women suffer from allergies somewhat more frequently than men, although the sex difference is reversed among children. Allergies occur in all age groups, with most studies showing that people in their twenties are most frequently affected. People with higher levels of education, in more highly qualified jobs and living in urban areas are more commonly affected by allergies than people from lower socio-economic levels and rural communities. The internationally identified increase in trend can also be identified in Austria with a 2fold, 3.6fold, and 4.6fold increase in the prevalence of hay fever, asthma and atopic eczema respectively, determined from the military health examinations of all recruits for national service between 1986 and 2003/04, although a clear decline in allergy prevalence was registered between 2003/04 and 2005. Health reports like the first Austrian Allergy Report provide the basis for international comparison of basic data. These data also enable the evaluation of the impact of different diseases on the health system as well as the development of public health strategies. PMID- 17915436 TI - Subjective health and impaired quality of life due to allergies in a representative population survey. AB - OBJECTIVES: To examine the quality of life among patients with different allergic diseases and to assess the health perception of patients with allergies compared to patients without allergies. METHODS: The source of data was the Vienna Health and Social Survey, a representative cross-sectional survey, commissioned by the City of Vienna. RESULTS: 18.7% of those suffering from allergies in the year before questioning reported that they had felt "very impaired", 30.0% felt "quite impaired", 42.4% "a little impaired" and 8.9% "not impaired at all". Women felt "very impaired" as a consequence of allergy more often than men (21.8% vs. 15.2%, p < 0.05). On a scale of 0 to 100 (0 = worst possible; 100 = best possible state of health) patients with allergies rated their state of health as distinctly worse than those people without allergies (71.7 vs. 74.1 points, p < 0.001). Of the various allergic illnesses patients suffering from asthma or allergy-related intestinal problems experienced the greatest impairment in quality of life. For these two diseases physical ability and feeling fit enough to do what one would like to do was also significantly reduced, when compared with subjects without allergies. CONCLUSION: Compared to people without allergies, patients with allergies have a significantly more impaired quality of life. It is important to take this into account when deciding upon a treatment regime. PMID- 17915437 TI - [Economic aspects of allergies: status and prospects for Austria]. AB - According to epidemiological studies there has been an increase of allergic diseases and corresponding costs. Health economics analyses intend to offer decision guidance towards a more efficient and effective resource distribution, in the conflicting relationship of medicine and economics. In analogy with the "Global Initiative for Asthma (GINA)-study", one arrives at costs of Euro 227.7 455.4 million for the Austrian health system in 2004 for asthma. Direct costs of allergies in general are in part available from health insurance carriers. Between 1998 and 2005 the number of nasal preparations and antiobstructive therapies prescribed rose by 15% and 50% respectively, whilst the costs of these drugs dramatically increased by 96% and 70% respectively. Expenditure on anti histamines rose by 31% between 2002 and 2004, whereas costs of topical and systemic anti-allergic drugs remained constant. Costs of allergies also include additional costs carried by the individuals affected, which must be added to those covered by the national health insurance carriers. Furthermore, patients with allergies more frequently turn to alternative and complimentary medicine to treat their condition (above all homeopathic remedies, massages and ointments) than people not suffering from allergies. Indirect costs due to allergies are, for instance, those caused by sick leave and loss of productivity. A continual systematic evaluation of available data on allergies in Austria could contribute to more effective implementation of medicines. PMID- 17915439 TI - [The influence of drug packaging on the drug-taking compliance of older patients living on their own]. AB - The present study is concerned with compliance and the possible influence thereon of drug packaging. Since most medications are prescribed for older persons, our target group was that of the over 70-year-olds otherwise usually excluded from participating as probands in clinical trials. Our method chose a study design that includes both the experience of a selected group of experts as well as the results of a field study. In the field study, 70 persons (18 male and 61 female), and 70-to-95 years of age participated in a face-to-face interview as well as in a test dealing with the handling of drug packaging. While the subjective patient responses provide a positive result with respect to compliance -- over 80% say that they take their medications regularly -- the data that were determined objectively within the framework of the test with selected medications showed deficiencies. The "medication test" carried out here shows impressively that child-proof packaging is also "age-proof" and, when used in the target group of older persons, can represent a considerable barrier to compliance. PMID- 17915440 TI - [Features of ontogenesis of uroliths]. AB - The aim of the study was investigation of urolith growth process using the method of ontogenetic analysis. A total of 435 autochtonic uroliths obtained after surgery or spontaneous elimination from 422 patients with urolithiasis were studied with roentgenophasic analysis on diffractometer DRON-3, IR-spectroscopy, electron-probe. The concrements incorporate several zones with a nucleus as one of the zones. In some cases spherolith consisted of several splitted individuums with different orientation. Their nuclei contained phosphates and organic substance resultant from renal inflammation. In the other cases lithogenesis of the spherolith started from monocrystal microscopic nucleus which arrived from the kidney. The number of spherolith emergences can be interpreted as a rhythmic course of the disease. A prolonged macrorhythm characterizes stable disease. Formation of cut surfaces of the spherolith defines interruption of rhythmic zonality. A change in the composition of the crystal medium does not stop concrement growth, it provokes replacement of one mineral phase for the other. The ontogenetic analysis has detected that in the position of the concrement in the kidney mineral substance of its deep zones undergoes phasic transformations and morphological alterations. In changing medium conditions there are recovery of surfaces of solution with a change of a mineral phase, formation of new growth rhythms, aggregation and compression of the particles. This explains mixed composition of most of uroliths. PMID- 17915438 TI - [Atypical antipsychotics and metabolic syndrome]. AB - The introduction of atypical antipsychotics in psychopharmacology represented a major advance in the treatment of psychotic disorders. However, there have been numerous studies that certain atypical antipsychotics may be associated with a greater risk of metabolic abnormalities than others, including weight gain, hyperlipidemia and new-onset typ 2 diabetes mellitus. A G-Protein beta3 subunit Gen (C825T) polymorphism, an increased carbohydrate metabolism and dyshormonism are discussed as pathogenetic mechanisms. High risk patients (adiposity, hyperlipidaemia, hyperglycaemia, preexisting diabetes) should maintain an antipsychotic agent with a favourable side effect profile. In these cases a periodical diabetes screening and blood lipid controls are required. Clinicans must balance the significant benefits of atypical antipsychotics against the risk of metabolic disturbances. In this article recent findings are reviewed. PMID- 17915441 TI - [Transurethral endoscopic pyelolithotripsy in combined treatment of complex forms of nephrolithiasis]. AB - The article presents results of retrograde endoscopic pyelolithotripsy (REP) in 75 patients with large and coral nephroliths. Retrograde contact pyelolithotripsy was made with application of Swiss Lithoclast unit, semirigid ureteroscopes R. Wolf with a cone 8-9 Ch and working channel 5.3 Ch. This enables maximal removal of the destroyed concrement by hydraulic litholapaxia with consequent insertion of the internal stent. Extracorporeal lithotripsy with low-energy shock-wave impulses was performed on demand (1-3 sessions). The stent was removed in 3-5 days in cases of complete one-stage sanation of the kidney and in 2-3 weeks in the presence of residual fragments of the nephrolith. One-stage complete sanation in coral nephrolithiasis (K1-K2) up to 5 cm3 and large concrements was achieved in 69.3% patients. Three months later residual fragments of the nephroliths were observed in 8% patients, 6 months later--in 5.3%. One year after treatment a complete sanation was achieved in 96%. Complications of the method consisted in exacerbation of chronic pyelonephritis observed in 12% cases. The control examination revealed improvement of renal function early and late after surgery. As shown by 6-year follow-up, recurrent nephroliths occurred in 6% patients. Endoscopic retrograde pyelolithotripsy can be used as a method of choice in the treatment of compound forms of nephrolithiasis in intrarenal type of the pelvis, the absence of marked extension of the caliceal-pelvic system of the kidney, in other stones resistant to extracorporeal lithotripsy. PMID- 17915442 TI - [Effects of magnetolaser radiation on pathologic process in the kidneys in experimental renal colic]. AB - The experiments were made on 22 rabbits with induced single or recurrent renal colic. Ten animals after renal colic were exposed to magnetolaser radiation. The histological examination of renal tissue demonstrated colic-related changes both in renal body corpuscle and ductules. Parenchimal edema, parabiotic processes in epitheliocytes and even necrosis of some cells were found. In case of recurrent colic the changes were severe, much more epitheliocytes were damaged, inflammatory leucocyte infiltration appeared. Magnetolaser radiation reduces the damage, eliminates leucocyte infiltration. Lymphocytic infiltrates formed in renal interstitium after two sessions of magnetolaser therapy indicate stimulation of cell immunity. The same processes develop also in the contralateral kidney. Their activity depends on the number of colics. Therefore, magnetolaser radiation has a protective action both on the isolated organ and body as a whole. PMID- 17915443 TI - [Use of levofloxacin in transcutaneous nephrolithotripsy]. AB - The aim of the study was elucidation of levofloxacin (tavanik) efficacy in transcutaneous nephrolithotripsy (TNLT) made to prevent or treat infectious inflammatory complications. We performed 211 transcutaneous endoscopic operations on the kidneys for urolithiasis (71 males and 117 females). Of them, 47 (25%) patients received tavanik. The concrement was located in the pelvis (n=9), calyx (n=9), pelvoureteral segment (n=5). Coral concrements were found in 24 cases (K1- in 12, K2--in 6, K3--in 3, K4--in 3 patients). Nephrolitholapaxy was performed in 39 patients, nephrolithoextraction--in 8 patients. Duplex ultrasound dopplerography conducted in all the examinees before surgery and on day 5-6 after it gave information about renal blood flow. Duration of the operation varied from 20 minutes in nephrolithoextraction to 136 minutes in nephrolitholapaxy of the coral concrements (mean time 40 +/- 6 min). Tavanik' was given according to the following scheme: 24 hours before the operation 500 mg tavanik was given orally, on the day of surgery and 3 consequent days 500 mg intravenously dropwise, the next 4 days 500 mg orally. Thus, a course of tavanik lasted 8 days. Nephroliths were removed in all the patients. Mean hospital stay was 7 days. In the group of 47 patients intraoperative complications were not observed. Pelvic microperforation (1 case) and loss of the concrement fragment in the paranephria (1 case) were not considered as complications and had no effect on postoperative period and recovery of the patient. Postoperative complications occurred in 6 (12.7%) patients: late hemorrhage (1), pyelonephritis (4), bacteriotoxic shock early after surgery (1). Thus, postoperative infectious-inflammatory complications arose in 5 (10.63%) patients. These patients received the same antibacterial treatment as the other patients except one patient with bacteriotoxic shock in whom a daily dose of tavanik was raised to 1 g in the course of 2 days. As success of transcutaneous nephrolithotripsy depends much on adequate pre- and postoperative antibacterial therapy, tavanik (levofloxacin) can be effectively used as monotherapy in transcutaneous nephrolithotripsy to prevent and treat postoperative infectious-inflammatory complications. PMID- 17915444 TI - [Retroperitoneoscopic ureterolithotomy]. AB - Ureterolithotomy has an alternative which is much less invasive - retroperitoneoscopic ureterolithotomy (RU). Nineteen cases of RU are reported (15 males, 4 females). Thirteen patients have undergone RU as monotherapy, 5 patients -after ineffective sessions of extracorporeal shock wave lithotripsy, 1 patient- after contact ureterolithotripsy. Mean size of the stones was 12.8 (9.5-30) mm. All the stones were eliminated for one procedure. Duration of surgery in the first 5 operations was 1.5-2 hours, in subsequent ones--65-70 min. The number of bed-days varied from 2 to 10 days (4.2 days). One male had a postoperative attack of pyelonephritis which was treated with antibacterial drugs. RU is a safe and effective operation, it is justified when previous surgery was ineffective, in large, longstanding concrements of the upper and middle third of the ureter, monohydrate-oxalate concrements. PMID- 17915445 TI - [Temporary and permanent NiTi stents in percutaneous surgery for ureteropelvic segment stricture]. AB - A total of 18 roentgenoendoscopic percutaneous interventions for strictures of the ureteropelvic segment (UPS) have been made by the authors from the beginning of 2004: in 15 (83.3%) females and 3 (16.7%) males aged 16-48 years. Recurrent strictures developed in 11 (61.1%) patients after operations by Kuchere, Pristlay, Andersen-Hines, in 7 (38.9%) patients the strictures were primary. UPS bougieurage and endotomy were made in 3 and 15 patients, respectively. UPS intubation with a PVC tube was made in 2 cases, with a permanent NiTi stent-crown -in 5 cases, temporary stent--in 9 cases, in 2 cases intubation was not made. Complete recovery of UPS passability and significant reduction of hydronephrosis were seen in 15 (83.3%) patients, insignificant effect was registered in 3 patients (one case of PVC tube intubation, one case of NiTi temporary stent, one case of no intubation). Efficacy in patients with permanent stent was 100%, in patients with temporary stent--88.9%, in patients with a PVC tube and without intubation--in 50%. Novel techniques of intubation with permanent and temporary stents with shape memory after percutaneous correction of UPS stricture are promising in the treatment of patients with this pathology of the upper urinary tract and may become an operation of choice. PMID- 17915446 TI - [Photodynamic therapy of prostatic adenoma]. AB - We were the first to study feasibility of photodynamic therapy (PDT) for prostatic adenoma using photosensitizer from chlorine group--photoditasine. We have developed and used PDT technique with application of 3-way silicon catheter with two balloons. This allows care for presenile and senile patients with compromised somatic status who are at high surgical risk of standard operations. The results demonstrate regression of both irritative and obstructive symptoms, improvement of life quality, persistence of sexual function. PDT reduces hospital stay as it can be used in outpatient conditions. PMID- 17915447 TI - [A variant of transvesical extraurethral adenomectomy]. AB - To facilitate extraurethral adenomectomy, we used a semicylindric scalpel and metallic tube (tubus) to make a semioval cut parallel to proximal prostatic portion of the urethra along the posterior semicircle of the bladder neck in 260 patients with prostatic adenoma (PA). The urethra was isolated from the adenomatous tissues. The vesicular cut was prolonged left and right to the level of ureteral ostia. This manipulation was followed with bimanual index finger enucleation of adenomatous tissue. The defect of the vesicourethral segment was sutured with catgut or monocril. Extra-urethral adenomectomy with semicylindric scalpel injured prostatic urethra in lesser extent than endourethral adenomectomy, it reduced the risk of hemorrhage, entailed neither stricture of the posterior urethra nor incontinence of urine in the postoperative period. PMID- 17915448 TI - [Complex therapy of irritative voiding disorders after transurethral resection of the prostate and adenomectomy]. AB - Rehabilitation therapy was performed in 5 groups (n=122) of patients operated for prostatic adenoma (TUR of the prostate, transvesical prostatectomy) and having postoperative marked irritative miction disorders. Group 1 received antibacterial therapy (AT); group 2-- AT+alpha-adrenoblockers; group 3-- AT+transrectal laser and magnetic physiotherapy; group 4-- AT+alpha-adrenoblockers+transrectal laser and magnetic physiotherapy; group 5-- AT+prostatotropic phytotherapy (gentos). The worst result was achieved in group 1, the best one--in groups 3 and 4. Thus, adjuvant physiotherapy inclusion in the complex of postoperative rehabilitation of patients operated for prostatic adenoma is justified as it improves treatment efficacy. PMID- 17915449 TI - [24-h dynamics of urination in patients with overactive urinary bladder]. AB - Home 3-day uroflowmetry was made in 41 female patients (mean age 46 years) before and after therapy for overactive bladder (OB). The course of therapy lasting from 6 weeks to 3 months included M-cholinolytics and vascular medication. Estimation of the bladder capacity in different time of 24-h period revealed circadian deviations of urinary flow and volumes. Administration of short-term M cholinolytics enables chronotherapy providing maximal effects within 4-6 hours, possibility to vary doses. In view of this, optimal cholinergic therapy of OB is trospium chloride which has necessary properties, high safety and wide dose range. PMID- 17915450 TI - [Male infertility in delay of sexual development]. AB - The examination of 260 infertile males revealed that delay of sexual development (DSD) predisposes to male infertility. DSD is often characterized by autoimmune reactions pointing to DSD as one of the causes of autoimmune infertility. Spermatozoa in DSD have low viability. Hormonal changes in DSD in adult males did not significantly change from the group with normal sexual development. LH/testosterone and testosteron/estradiol differed significantly in DSD and hypogonadism. Calculation of these proportions may serve differential criteria of DSD and secondary hypogonadism. PMID- 17915451 TI - [The role of testosterone drugs in combined therapy of erectile dysfunction in patients with metabolic syndrome]. AB - The aim of the trial was assessment of efficacy of combined therapy with testosterone drug (androgel) and wardenafil in patients with erectile dysfunction (ED) and metabolic syndrome (MS). The trial included 16 males with organic ED, laboratory and clinical symptoms of hypogonadism and MS (mean age 63.8 +/- 8.4) who had failed monotherapy with phosphodiesterase of type 5 (FDE-5). Both routine methods and special methods of ED and MS diagnosis with application of International Index of Erectile Function were used in examination. MS was diagnosed according to the criteria of the National Cholesterol Educational Program. Wardenafil was given in a dose 20 mg 1 hour before coitus, but not less than 4 tablets a month. Androgel applications were given in a daily dose 50 mg. The treatment lasted for 3 months. The combined treatment significantly reduced body mass index, waist circumference, improved erectile function and libido. Erectile function normalized in 3 (18.75%) patients. Carbohydrate and lipid metabolism improved, the levels of total and free testosterone normalized. Elevation of sexual hormones concentration eliminated clinical hypogonadism symptoms in 68.75% patients. Thus, combined treatment with FDE-5 inhibitor wardenafil and testosterone drug androgel is effective, safe, improves erectile function, hormonal, lipid and carbohydrate blood profile and can be used in patients with MS, ED and hypogonadism. PMID- 17915452 TI - [Correction of local and central disorders in patients with chronic abacterial prostatitis in usage of amus-01-intramag unit]. AB - Combined use of dynamic magnetotherapy of local and central action in combination with antibacterial endourethral therapy and rectal administration of vitaprost arrests symptoms of chronic prostatitis. Combination of local and total dynamic magnetotherapy with application of AMUS-01-INTRAMAG unit improves erection quality in 81.8% males with psychogenic form of erectile dysfunction. There was also improvement of spermogram parameters and relieve of asthenovegetative syndrome. PMID- 17915453 TI - [Estimation of predictive prostate cancer probability with logistic regression equation]. AB - Low specificity of PSA for early diagnosis of prostate cancer (PC) is the cause of search for new tests. The aim of our study was to develop the logistic regression model and estimate the value of the regression equation as a diagnostic tool for prostate cancer detection. A total of 518 male patients aged 47-83 years (mean 65.5 +/- 6.5 years) who had undergone TRUS-guided 12-core systematic transrectal prostate biopsy were included in the study. PC detection rate in our study was 43.8%. The logistic regression model with PC detection as a response and age, prostate volume, PSA, induration on DRE and hypoechoic lesion on TRUS as effects was designed. With regression equation PC probability for any patient was calculated. The regression equation was tested as a PC diagnostic tool. As the combination of model effects (chi-square 87.9; p < 0.0001; R2 = 0.124) any of the effects independently may predict prostate cancer detection. The obtained regression equation is: P(Pca) = 1/{1 + 2.718(-[-4.029 + (0.068 x AGE) + (0.022 x PSA) + (-0013 x PROSTATE VOLUME) + (0.375 x DRE) + (0.254 x TRUS)])} Accuracy (area under ROC-curve) of our regression equation as a PC detection diagnostic tool was 73%. Probability cutoff of 0.26 leads to sensitivity of 90% and specificity of 30% and eliminates 12% of unnecessary biopsies in patients with benign prostate diseases (chi-square 10.91; p < 0.0001). Thus, the obtained logistic regression equation may be used as a PC diagnostic tool in the suspects. Multicenter trial may improve regression equation diagnostic performance. PMID- 17915454 TI - [Physiological and pathological conditions of the prepuce in childhood: treatment methods]. AB - The authors analyse treatment of 3235 patients with prepuce pathology aged from 8 days to 16 years, the results of the treatment of preputial sac pathology; describe clinical manifestations of phimosis (physiological, hypertrophic, cicatricial) in children; formulate treatment policy, indications and contraindications for conservative and surgical treatment; provide practical recommendations on hygiene and care for infant and young age boys. PMID- 17915455 TI - [Lymphofibromatosis of the penis and scrotum]. PMID- 17915456 TI - [Foreign body of the kidney due to migration of the toothpick from the intestine to the kidney]. PMID- 17915457 TI - [Effective therapy in combined treatment of acute purulent pyelonephritis in an early postoperative period]. PMID- 17915459 TI - [Implementation of the municipal special tuberculosis control program and its impact on the epidemic situation]. AB - The implementation of regional tuberculosis control programs contributes to the improvement of the tuberculosis epidemic situation. The holding of a stimulating lottery among those who have undergone fluorographic study is a novel approach that increases the coverage of particularly the unorganized population with preventive fluographic studies. Social support of patients with tuberculosis improves the results of their treatment. To organize sanitary educational work among the population promotes the increase of its awareness and hence the improvement of preventive measures against tuberculosis. PMID- 17915460 TI - [Drug-resistant tuberculosis in the Republic of Karelia]. AB - The study of drug resistance (DR) trends in the Republic of Karelia (2000-2005) indicated an increase in the incidence of multidrug resistant (MDR) tuberculosis. Among the contingents of patients, there was a particularly considerable increase in MDR tuberculosis from 18.4 to 57.6%. Analysis of the DR spectrum showed that the rate of DR to a combination of three first-line drugs was 65.3% in 2005. Retrospective analysis of 67 case histories of MDR tuberculosis revealed that in 23 (34%) patients the cause of multidrug resistance was a DR rise during therapy (amplification). Thus, one of the most important causes of an increase in the incidence of MDR tuberculosis among the contingents is a wide spread of multidrug resistance (to 3 drugs or more - 65.3%) during which the use of chemotherapy IIA (2S, H, R, Z, E + 1H, R, Z, E) results in the amplification of DR and the development of multidrug resistance among the contingents. PMID- 17915461 TI - [Comparative evaluation of the informative value of the rapid serochek mbt test to determine mycobacterium tuberculosis antibodies and mantoux tuberculin test in adults]. AB - The diagnostic informative value of the Serochek MBT test (Zephyr Biomedicals, India) in detecting serum Mycobacterium tuberculosis antibodies was compared with that of the Mantoux tuberculin test with 2 TE of PPD-L in patients with pulmonary tuberculosis. The informative value of the study rapid method for the diagnosis of tuberculosis was generally 1.8 times less than that of the Mantoux tuberculin test with 2 TE of PPD-L (only intensive and hyperergic tuberculin tests were kept in mind): 6.1 times less in patients with small forms; 1.4 times less in the examinees with disseminated processes; 1.5 times greater than in chronic pulmonary tuberculosis. Since the detection and diagnosis of tuberculosis are very important at an early development stage of the disease, the findings (9% positive serotests in small forms of pulmonary tuberculosis) suggest the low sensitivity of the approved Serochek MBT test in verifying the presence of serum Mycobacterium tuberculosis antibodies and therefore its low informative value as a method to be used for the rapid diagnosis of tuberculosis. PMID- 17915462 TI - [Role of the abnormal tracheobroncheal tree in the efficiency of treatment of patients with pulmonary tuberculosis]. AB - The efficiency of treatment has been studied in 100 patients with pulmonary tuberculosis caused by drug-resistant (DR) Mycobacterium tuberculosis (MBT) strains and complicated by bronchial diseases, treated at the units of the Clinic of Phthisiopulmonology, I. M. Sechenov Moscow Medical Academy, in 2004 to 2006. Several forms of specific and/or nonspecific bronchial diseases are shown to be detected in patients with drug resistance in MBT. Pulmonary tuberculosis caused by DR MBT strains and complicated by bronchial diseases is characterized by the lower interferon-producing capacity of bronchoalveolar lavage (BAL) cells and by more significant impairments in the mechanisms of local lung protection. Inclusion of the immunomodulator leukinferon into the combined therapy of patients with pulmonary tuberculosis complicated by bronchial diseases contributes to the elevation of BAL cell production of alpha and gamma interferons, reduces the generation of active oxygen forms in the BAL phagocytes and brush biopsy specimens, and increases the levels of immunoglobulins in BAL. The local administration of leukinferon reduces the time of cure of specific and nonspecific bronchial diseases in patients with pulmonary tuberculosis who isolate DR MBT strains, thus promoting the enhanced efficiency of treatment in reducing the times of bacterial isolation and decay cavity closure and the length of hospital stay. PMID- 17915463 TI - [Principles in the performance of chemotherapy in dispensary group VI children and adolescents]. AB - Based on the annual reports of Moscow tuberculosis dispensaries, the analysis of their follow-up of dispensary registration group VI patients, the authors have developed preventive chemotherapy regimens in relation to the specific features of tuberculin sensitivity, the presence or absence of additional risk factors (concomitant somatic diseases, social risk factors). Regimen I (1 antituberculous drug (isoniazid) for 3 months) may be used in dispensary registration groups VIA and VIB patients without additional risk factors. Regimen II (2 antituberculous drugs (isoniazid + pyrazinamide/ethambutol) for 3 months) may be employed in dispensary registration groups VIA and VIB patients with additional risk factors, as well as in children from dispensary registration group VIB, whether additional risk factors are present or absent. A repeated course of preventive chemotherapy regimen II is indicated when tuberculin sensitivity is increased or a hyperergic reaction is retained after termination of treatment in patients having additional risk factors. PMID- 17915464 TI - [Multislice computed tomography in the diagnosis of nephrotuberculosis]. AB - The sensitivity of the conventional radiotherapy involving excretory urography and ultrasound scanning is 80.3%. The clinical and X-ray form of new-onset nephrotuberculosis is being specified and revised during a follow-up and, in some cases, intraoperatively or at autopsy. The main reason for discrepancy between preoperative and postoperative diagnoses is inadequate visualization of the upper urinary tract when standard radiation diagnosis is used. Multislice computed tomography makes it possible to visualize the renal parenchyma, its vessels and urinary tract and yields a real three-dimensional image of urinary organs, to ascertain the pattern of urinary tract abnormalities, the preservation of the renal vasculature, and the functional reserves of the diseased and contralateral kidneys, and to define the only correct management tactics for patients with nephrotuberculosis in 93.3% of cases. PMID- 17915465 TI - [The clinical and social characteristics of the contingents of patients with respiratory tuberculosis during the sanatorium-stage treatment]. AB - The specific features of contingents of patients with respiratory tuberculosis were studied during the sanatorium-stage treatment in 2004-2005. Out of 317 patients (57.4% females and 42.6% males), 70.7% of the patients aged under 45 years; the females were more frequently observed to have focal forms of the disease whereas the males had infiltrative forms and were more frequently operated on for tuberculosis. 27.1% of the patients (more commonly males) had underweight (more than 10 kg body weight deficiency); 36.3% were unemployed; the women's social status was higher than the men's; 72.5% of the patients had comorbidity. Drug intolerability was observed in 21.8% of the patients and 20.5% did not complete treatment. The above features complicate a rehabilitative process and require a significant correction of the treatment recommended by dispensaries, as well as additional therapeutic and diagnostic measures. PMID- 17915466 TI - [Pulmonary function in patients with infiltrative pulmonary tuberculosis]. AB - Vital capacity (VC), forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), FEV1/VC%, PEF, MEF25, MEF50, MEF75, TLC, TGV, pulmonary residual volume (PRV), R(aw), R(in),, R(ex), DLCO-SB, DLCO-SS, PaO2, and PaCO2 were determined in 103 patients with infiltrative pulmonary tuberculosis. Pulmonary dysfunction was detected in 83.5% of the patients. Changes were found in lung volumes and capacities in 63.1%, impaired bronchial patency and pulmonary gas exchange dysfunction were in 60.2 and 41.7%, respectively. The changes in pulmonary volumes and capacities appeared as increased PRV, decreased VC and FVC, and decreased and increased TGV and TLC; impaired bronchial patency presented as decreased PEF, MEF25, MEF50, MEF75, FEV1/VC% and increased R(aw) R(in), and R(ex); pulmonary gas exchange dysfunction manifested itself as reduced DLCO-SB, DLCO-SS, and PaO2 and decreased and increased PaCO2. The magnitude of the observed functional changes was generally slight. Significant disorders were observed rarely and very pronounced ones were exceptional. PMID- 17915468 TI - [Efficiency of a combination of haloaerosols and helium-neon laser in the multimodality treatment of patients with bronchial asthma]. AB - A hundred and thirty-eight patients with infection-dependent bronchial asthma, including 73 with moderate persistent asthma and 65 with severe persistent one, were examined. Four modes of a combination of traditional (drug) therapy (DT) and untraditional (halotherapy (HT) and endobronchial helium-neon laser irradiation (ELI) one were used. The efficiency of the treatment performed was evaluated, by determining the time course of clinical symptoms of the disease on the basis of scores of their magnitude and the patients' condition. The findings indicated that in moderate persistent asthma, both HT and ELI in combination with DT exerted an equal therapeutic effect, which provided a good and excellent condition in 83.3% of cases. In severe persistent asthma, such a condition was achieved in 93.75% of cases only when multimodality treatment involving DT, HT, and ELI had been performed. PMID- 17915467 TI - [The specific features of formation of a humoral immune response in patients with tuberculosis]. AB - The paper presents the results of studying the time course of changes in tuberculosis antibodies by enzyme immunoassay (EIA) and immune blotting (IB) in 34 patients with pulmonary tuberculosis. In 76.5% of cases, EIA were established to reveal tuberculosis antibodies whose concentration is most pronounced in lung tissue destruction. The detection rate of specific antibodies to Mycobacterium tuberculosis (MBT) structural components by IB was 100%, which was 23.6% greater than that by EIA. The MBT structural components with a molecular weight of 11-20, 21-30, and 101-160 kDa were found to have high immunogenic properties, as suggested by the maximum levels of antibodies and by the marked antigenic proteins of proteins with a molecular weight of 11-20, 21-30, and 41-50 kDa, which showed itself as the generation of the maximum levels of circulating immune complexes with antigens of this specificity. PMID- 17915469 TI - [The branches of academician S. N. Fyodorov eye microsurgery complex: experience and prospects]. AB - The article is dedicated to the history of the creation of the branches of Eye Microsurgery Complex (EMC), the period of their establishment, and their present state. The article demonstrates the big part played by EMC in providing ophthalmologic aid in the country. The development of the therapeutic/diagnostic network around the branches makes high-quality medical aid available for the population, which is a part of national health project. The therapeutic/diagnostic base of the branches, their scientific and teaching work as well as high professionalism of the personnel make the branches leaders in the system of the organization of ophthalmological aid and rendering high technological aid in regions. PMID- 17915470 TI - [The problems of cataract surgery and intraocular correction: Russian school achievements and modern trends]. AB - The development of the safest and, at the same time, efficient methods of cataract surgery, is not only a topical task of medical science and practice, but also, taking the demographic situation into account, a solution to a social problem. The article describes the main achievements of academician S. N. Fyodorov's school in the field of cataract surgery with intraocular lense (IOL) implantation. Special attention is paid to the key components of cataract extraction technique named "small incision surgery", which is based upon the use of ultrasmall surgical approaches with self-sealing ability and no need for suturing. The main advantages and disadvantages of various types of cataract lense fragmentation, including ultrasound emulsification, mechanic and hydraulic fragmentation, and laser destruction, are described. Topical issues concerning cataract extraction technique under the conditions of complicating ophthalmopathology including the weakness of ciliary zonule system, high-degree myopia, glaucoma, pseudoexfoliative syndrome, small pupil, past injuries and eye surgery, are outlined. The paper also reflects a new qualitative step in the development of intraocular aphakia correction technique under modern conditions. Basic requirements to design parameters and IOL materials are determined from the position of reestablishing the accommodative function and fine visual functions. Ways of IOL design improvement are described especially for their application in non-standard and complicated cases. PMID- 17915471 TI - [Achievements of S. N. Fyodorov's school in the field of keratoplasty and cornea replacement]. AB - Reduction in corneal transparence leading to the loss of sight may be caused by different factors, such as eye burns and injuries, keratitis, corneal ulcers, primary and secondary dystrophy, keratoconus, and keratoglobus. In many cases, keratoplasty presents an effective treatment. S. N. Fyodorov made a great contribution to the development of cornea replacement. This was him who placed the entire ophthalmology, including keratoplasty, at the microsurgical level, suggested the use of large-diameter grafts, and was the initiator of establishing the first Eye Tissue Bank. Perfection of microsurgical technique, selection and protection of the graft, as well as new approaches to pre- and postoperation therapy have provided transparent engrafting in 35 to 95%. Presently, Eye Microsurgery Complex is the only Russian institution where the problem of cornea replacement is being worked out as a scientific and practical issue. Three models of keratoprosthesis have been developed: Fyodorov-Zuyev model for implantation into a burn leukoma, Moroz-Glazko model for implantation into a dystrophic leukoma, and a "grid" for implantation into a leukoma of any etiology. PMID- 17915472 TI - [The role of eye tissue bank in cornea transplantation]. AB - Cadaveric human cornea transplantation is impossible without a specialized tissue donor service, Eye Tissue Bank. An original medical technological system of Eye Tissue Bank was developed at Fyodorov Eye Microsurgery Complex, a federal state institution. The system is still unique in the world; it is grounded upon the concept of the pharmacological protection of the graft and is oriented towards prevention of energodeficiency both in donor cornea and in the recipient's organism at all stages from the selection and preservation of cadaveric material till postoperative pharmacological correction of the recipient. The working experience of Eye Microsurgery Complex has shown the significance of the developed medical technological system for clinical ophthalmology and rehabilitation of the blind and patients with impaired vision in Russian Federation. PMID- 17915473 TI - [Contemporary achievments in ophthalmotraumatology]. AB - The paper presents the results of research work carried out in Eye Trauma Department during the last 30 years, covering in particular new issues in penetrating wound management and astigmatism prevention, original methods of foreign body removal, some approaches to eye transplantation, the pathogenesis and treatment of eye burn disease, surgical treatment of close eye injury, and research performed in collaboration with the center of disaster medicine Zaschita. PMID- 17915474 TI - [Possibilities for prevention and treatment of blindness and impaired vision in children caused by congenital eye diseases]. AB - Blindness, disability, and impaired vision present important medicosocial problems. According to the WHO, there are 1.5 million blind children in the world. The prevalence of child blindness in Russia is 1.6, and that of impaired vision is 3.5 per 10000 children. It is considered that child blindness can be prevented in 40 to 50% of children. According to data collected during ophthalmological examinations in specialized school, blindness in 88 to 92% of cases is caused by. Presently, significant achievements have been made in treatment of congenital eye diseases. Application of modem surgical techniques in patients with cataract, aphakia, or amblyopia makes it possible to increase visual acuity to 0.3 or higher in 35% of children, and to 0.005 to 0.25 in 45% of children. Modem pathogenetically directed surgery together with timely treatment allows for the normalization of intraocular pressure and visual stabilization in 76.4 to 86.4% of children with congenital glaucoma. In this connection, the achievements of scientific research should be introduced into practice. Realization of the 1999 WHO program on liquidation of eliminable blindness include three key directions: treatment and prevention of blindness; consolidation of the infrastructure and technology of ophthalmological aid; training of specialists. PMID- 17915475 TI - [Prognostication of the results of treatment of retinal detachment with silicone oil tamponade using pattern recognition methods]. AB - The aim of this work was to solve the problem of prognostification of anatomic and functional results of treatment of retinal detachment (RD) with silicone oil tamponade. Analysis of the treatment of 257 eyes was performed using pattern recognition methods. All the patients were characterized according to 32 signs related the preoperative status only. As a result, an informative sign system was developed; the present data proved to be not representative enough, which manifested mostly in the class of RD recurrence, where the rate of correct end result recognition was 75% (in the class of "normal condition" it was 95%). One of the most significant RD signs, PVR degree, in this study was placed in the second ten according to its informative value, probably due to the fact that PVR classification is not detailed enough. PMID- 17915476 TI - [Laser thermokeratoplasty: clinico-functional aspects of postoperative eye condition]. AB - Laser thermokeratoplasty (LTK) induces postoperative inflammation of the cornea. Its clinical manifestation as well as the degree of endothelial cell loss depend on laser irradiation energy and the number of coagulates. The refractive effect of LTK is most stable in a hyperopia of up to 3.0 D and a laser energy value of up to 170 mJ; in this case, endothelial cell loss dose not exceed 2.2%. According to biomicroscopy, ultrasound pachimetry, and optical coherent tomography of the anterior segment of the eye, LTK results in corneal thickening, after which the volume of the coagulant gradually decreases and stabilizes by the end of the first year after LTK. Two types of scar, biomicroscopically different in size and optical density, form after LTK. The authors distinguish four clinical periods of healing after LTK, which makes it possible to evaluate and systematize physiological and biomechanical processes taking place in the cornea during refraction formation. PMID- 17915477 TI - [Organ-sparing treatment of uveal melanomas: safety borders]. AB - The history of organ-sparing treatment of uveal melanomas counts almost 40 years. During the last years, the development of new appliances and tools have widened possibilities provided by eye interventions, including both surgery and the application of various physical factors on eye tissues. However, when a malignant tumor, such as chorioid melanoma, is the case, there is a question of safety borders both for the patient (first of all) and the eye. On the basis of world experience and her own broad one, the author analyses the reasons for errors in treatment of such patients, and determines both efficiency and safety borders. PMID- 17915478 TI - [Photodynamic therapy in treatment of chorioid neovascularization]. AB - The authors studied the effectiveness of photodynamic therapy (PDT) with Photosence, a Russian photosensitizer, in treatment of chorioid neovascularization (CNV) in cases of age-related macular degeneration (ARMD) and pathological myopia (PM). The subjects were 73 patients with CNV suffering from ARMD and PM. The efficiency of PDT and complex conservative therapy was compared using vision acuity measurement, retinal morphometry, and fluorescent eye ground angiography (FEGA), performed before treatment, immediately after treatment, and 1, 3, 6, and 12 months later. The study showed that PDT in patients with CNV, ARMD and PM was more efficient than pharmacotherapy. Vision acuity improved or stabilized, and the parameters of retinal morphometry and FEGA improved as well. The results of the study evidence high efficiency of PDT with Photosence in treatment of CNV with ARMD and PM. PMID- 17915479 TI - [Diagnostics of ischemic ocular lesions in carotid arterial pathology]. AB - The aim of the study was to evaluate blood flow in ocular vessels and the carotid arteries of patients with various clinical forms of ischemic ocular lesion (IOL). The subjects were 165 patients with IOL aged 35 to 70 years. Of these, in 115 patients the lesion was acute, and in 50 patients it was chronic. Ultrasound examination using Doppler color mapping (DCM) and energetic mapping (EM) was performed to evaluate ocular and carotid blood flow. The use of DCM and EM made it possible to detect hemodynamically significant changes in acute and chronic IOL in carotid arterial pathology. Duplex scanning of the carotid arteries allowed for timely diagnostics of occlusive-stenotic carotid arterial lesion and determination of atherosclerotic plaque type. DCM and EM are highly informative methods of diagnostics of pathological vascular changes in IOL. PMID- 17915480 TI - [Development of a technique for ametropia correction with phakic intraocular lenses]. AB - A technique to correct ametropia with phakic intraocular lenses (PIOL) was developed on the basis of the analysis of anatomic, clinical, and functional condition of 409 eyes of 274 patients with various types of refraction, and operations performed on 279 eyes with myopia or hyperopia followed up for 2 to 10 years. Its algorithm is a step-by-step system including: patient selection based on a comprehensive clinical and functional examination with an obligatory ultrasound biomicroscopy and psychological testing; choice of PIOL design taking into account initial anatomic and functional parameters of the eye; simulation of the surgical effect using contact lenses allowing for calculation of the optimal PIOL power; choice of an adequate anesthesia method; direct PIOL implantation technique, which, when necessary, is combined with a simultaneous correction of astigmatism and strabismus; pre- and post-operation pleoptic treatment in cases of amblyopia of various degrees. PMID- 17915481 TI - [Mediators of the parameter constancy preservation in the local intercellular environmental pools, the differential nephron segmental function, and arterial hypertension]. PMID- 17915482 TI - [Plasma malonic dialdehyde levels in patients with recurrent erysipelas]. PMID- 17915483 TI - [Time course of changes in clinical and hematological parameters, iron metabolism, and free radical lipid oxidation in patients with iron-deficiency anemia during multimodality treatment]. AB - Isolated iron therapy and combined therapy including the antioxidant beta carotene were studied for their impact on the time course of clinical and laboratory changes and the state of free radical oxidation in patients with iron deficiency anemia. PMID- 17915484 TI - [Functional status of neutrophils in patients with autoimmune thyroiditis]. AB - The paper discusses the functional activity of peripheral neutrophilic granulocytes in 25 patients with autoimmune thyroiditis (AIT). The course of the latter is shown to be attended by a reduction in serum lactoferrin levels with simultaneous increases in the production of active oxygen forms and in the blood count of CD95+ granulocytes. PMID- 17915485 TI - [Cytochemical characteristics of neutrophilic granulocytes in various types of ischemic heart disease]. PMID- 17915486 TI - [Mycoplasmas in non-gonococcal urethritis]. AB - The paper presents data on the etiological role of different types of genital mycoplasmas, such as U. urealyticum (U), M. hominis (Mh), M. genitalium (Mg), in the development of non-gonococcal urethritis (NGU). A hundred and twenty patients with acute and chronic urethritis were examined. A control group comprised clinically healthy males. Polymerase chain reaction and cultural tests more frequently revealed genital Mycoplasmas in patients with acute and chronic urethritis than in the controls, Uu being more common than other mycoplasmas. Uu antibodies were detected only in some part of the infected, more frequently in acute urethritis. Class G antibodies to Mh were equally identified in healthy individuals and patients. Criteria for the involvement of Mycoplasma infection in the development of NGU are presented. PMID- 17915487 TI - [Approaches to species-specific typing of double mixed cultures comprising pseudotuberculosis bacteria and atypical plague bacillus strains]. AB - The species relevance of atypical Yersinia strains was determined by various microbiological, immunological, and genetic (including polymerase chain reaction) tests. These strains were shown to represent mixed cultures of Y. pseudotuberculosis serovariant O1b and Y. pestis var antiqua. Identification resistant cells with atypical properties and plasmid segregation were found in the populations of Y. pestis strains. Analysis of different diagnostic tests revealed the most reliable ones selected for the identification of atypical Y. pestis strains with unstable genome. PMID- 17915488 TI - [Clinical symptoms and treatment of cervicofacial actinomycosis. Literature survey and case report]. AB - Actinomycosis is a chronic, specific inflammation which is characterized by suppuration, abscess formation, tissue fibrosis and granuloma formation. Actinomycosis has three main forms (cervicofacial, which is the most frequent, approximately 60%, pulmonary and abdominal), but other regions of the body can be involved, too (e.g. neck, ovaries, bones), that is why its differential diagnosis becomes more and more relevant. Regarding its treatment, the majority of authors recommends the combination of surgical and antibiotic treatment. The authors of this article present a typical case of cervicofacial actinomycosis, in which the authors used the combination of surgical and antibiotic treatment. As a result of the treatment the healing process was completed successfully and without complications. PMID- 17915489 TI - [Significance of infection control in dentistry: a review]. AB - Dental care is a field of high priority regarding the risk of infections. Since many carriers are not aware of their infection, it may happen that the dentist meets a patient, in whom an earlier infection can be proven by serology, but the patient is not aware of it and the clinical signs and symptoms are missing, as well. For this reason, the dentist has to consider every patient potentially infected. On the other hand, health-care workers are not only susceptible persons to infections but they can also be sources of infections. In order to prevent the nosocomial infections the dentist has to ensure the hygienic protection of both the patients and the health-care workers. All the health-occupational measures have to be known and have to be kept by the dental personnel. The health personnel has to be informed on the risk and how to prevent infections. The essential importance of hygiene, the role of protective equipment and all the duties connected with should be emphasized. Furthermore, the continuing education of health-care workers is indispensable regarding the infectious diseases. In order to reduce the risk of nosocomial infections the authors summarize the state of-the-art knowledge of infection control. PMID- 17915490 TI - [Flow rate of minor salivary glands in elderly patients wearing complete dentures]. AB - Minor salivary glands show diverse levels of secretion in different regions of the oral cavity. The smallest production can be measured at the palatal glands, the highest in the buccal and the lingual glands. The labial glands show an intermediate value between the palatal and the buccal flow rate. According to the literature, secretion of the minor salivary glands decreases with age, yet only few data are available regarding the influence of removable dentures on the flow rate. The aim of this study was to assess the influence of complete dentures on the palatal, the labial and the whole saliva flow rate on elderly patients. A further aim was to compare the results obtained by the two measuring methods (weighing method and the PERIOTRON method), used to determine the flow rates of the minor salivary glands. According to the results of this study neither whole resting saliva flow rate nor the flow rate of the minor salivary glands (palatal, buccal) was influenced by long term removable denture wearing (denture wearers and controls -- weighing method: palatal: right side 2.4 +/- 3.3 microl/min/cm2, left side 1 +/- 3.8 microl/min/cm2 and labial: 1.4 +/- 2.6 microl/min/cm2; controls: right side 3.7 +/- 5.2 microl/min/cm2, left side 1.4 +/- 2.5 microl/min/cm2 and labial 1.8 +/- 3.9 microl/min/cm whole resting saliva flow: 0.32 +/- 0.26 ml/min and 0.29 +/- 0.24 ml/min respectively) (The data of denture wearers and controls -- PERIOTRON method: palatal right side 4 +/- 4.6 microl/min/cm2, palatal left side 3.5 +/- 3.6 microl/min/cm2 and labial 0.9 +/- 0.6 microl/min/cm2; controls: palatal right side 2.2 +/- 3.1 microl/min/cm2, palatal left side 1.8 +/- 1.8 microl/min/cm2 and labial 1.9 +/- 3 microl/min/cm2). Authors could not show difference between the weighing method and the PERIOTRON method applied in the measurement of the saliva flow rate of the minor glands. PMID- 17915491 TI - [Hungarian dentists' corporate response to eliminating mandatory membership in the Medical Chamber]. AB - Hungary's recent parliamentary act (Act XCVII. 2006) revoked the mandatory membership in medical chambers as a prerequisite of professional activity in the health service. After 31st of March 2007, members were automatically canceled except having addressed a unilateral declaration to the presidency of the relevant chamber from 1st of January until the above deadline. Dentists and physicians were mandatory members of the Hungarian Medical Chamber. According to the data obtained and analysed from the registry of this Chamber, out of all former dentist members (N = 5878) 5022 (85.44%) volunteered for the reorganized corporation. In cases of immigrant dentists (N = 522, 8.9% of the whole community) nine out of ten decided the same way (88.5%). Using specific distributions and correlations (gender, age groups, geographic location, practices on public financing) this study revealed many remarkable behavioural patterns of dentists' community to the changed legislation. PMID- 17915492 TI - Learning disabilities and difficulties: an Australian conspectus--introduction to the special series. PMID- 17915493 TI - Learning disabilities: bringing fields and nations together. AB - This article advocates an approach to supporting students who experience difficulties in learning, irrespective of nosology, particularly in the key areas of literacy and numeracy. In the state of Queensland, Australia, a distinction has been made between students' experiencing learning difficulties and those who have learning disabilities (LD). However, government priorities for improved achievement in literacy and numeracy have focused schools on the performance of all low-achieving students, without regard to diagnostic category. Many are now mobilizing a schoolwide effort that combines resources into a unified plan, using a three-wave approach. The first wave is high-quality classroom teaching, the second is early intervention, and the third is ongoing support for those students who have persistent difficulties, using adapted instruction and intensive tutoring. A further theme is the promise of neuropsychological advances for giving meaning to the underlying impairments of some students--who do have LD- that justifies the provision of adaptations to sustain their learning throughout their schooling and beyond. Throughout this article, the different yet converging understandings of LD in Australia and the United States are tracked, with suggestions made for future research that avoid the problems of operationalizing the definition of LD proposed by Keogh in 1982. PMID- 17915494 TI - Language delays, reading delays, and learning difficulties: interactive elements requiring multidimensional programming. AB - Researchers have hypothesized four levels of instructional dialogue and claimed that teachers can improve children's language development by incorporating these dialogue levels in their classrooms. It has also been hypothesized that enhancing children's early language development enhances children's later reading development. This quasi-experimental research study investigated both of these hypotheses using a collaborative service delivery model for Grade 1 children with language difficulties from a socially and economically disadvantaged urban community in Australia. Comparing the end-of-year reading achievement scores for the 57 children who received the language intervention with those of the 59 children in the comparison group, the findings from this research are supportive of both hypotheses. The interrelationships between learning difficulties, reading difficulties, and language difficulties are discussed along with children's development in vocabulary, use of memory strategies and verbal reasoning, and the need for multidimensional programming. PMID- 17915496 TI - Developing mathematical proficiency in the Australian context: implications for students with learning difficulties. AB - The teaching of mathematics in Australian schools has received considerable attention over the past decade. States and territories have designed and implemented new mathematics syllabi, and education sectors have supported teachers through sustained professional learning initiatives. Whereas the major focus of these initiatives has been on students constructing mathematical knowledge from a range of learning activities and quality instruction, the education of students with difficulties in learning mathematics has been given little direct attention. This analysis of current syllabus developments and professional learning initiatives highlights the risk of exposing students with learning difficulties to fragile program designs and classroom instruction. The research literature is also examined to propose ways in which these fragilities can be addressed. PMID- 17915495 TI - QuickSmart: a basic academic skills intervention for middle school students with learning difficulties. AB - QuickSmart is a basic academic skills intervention designed for persistently low achieving students in the middle years of schooling that aims to improve the automaticity of basic skills to improve higher-order processes, such as problem solving and comprehension, as measured on standardized tests. The QuickSmart instructional program consists of three structured, teacher- or teacher aide directed, 30-minute, small-group lessons each week for approximately 26 weeks. In this study, 42 middle school students experiencing learning difficulties (LD) completed the QuickSmart reading program, and a further 42 students with LD took part in the QuickSmart mathematics program. To investigate the effects of the intervention, comparisons were made between the reading and mathematics progress of the intervention group and a group of 10 high-achieving and 10 average achieving peers. The results indicated that although the standardized reading comprehension and mathematics scores of QuickSmart students remained below those of comparison students, they improved significantly from pretest to posttest. In contrast, the standardized scores of comparison students were not significantly different from pretest to posttest. On measures of response speed and accuracy gathered using the Cognitive Aptitude Assessment System (CAAS), QuickSmart students were able to narrow the gap between their performance and that of their high- and average-achieving peers. Implications are drawn regarding the importance of interventions that emphasize the automaticity of basic academic skills for students with learning difficulties. PMID- 17915497 TI - Teachers' in-flight thinking in inclusive classrooms. AB - This article explores the thinking of five junior high school teachers as they teach students with learning difficulties in inclusive classrooms. Insights into the ways these teachers think about students in these inclusive secondary school contexts were obtained through triangulating data from semistructured interviews, stimulated recall of in-flight thinking, and researcher field notes. Exploration of teachers' in-flight thinking (i.e., the thinking of teachers as they engaged in classroom teaching) revealed a knowledge of individual students that was not related to categorical notions of learning difficulties. This research has implications for the practice of teaching in inclusive settings as well as for teacher preparation. Specifically, it suggests that attention to student differences should be replaced by the development of teachers' knowledge about individual students as a rich source of practical knowledge and the basis for developing effective instructional techniques. PMID- 17915498 TI - Learning disabilities in Australian universities: hidden, ignored, and unwelcome. AB - Although more students with learning disabilities (LD) are enrolling in Australian universities, their learning needs are not well understood. This article reports on the experiences of students with LD who are encouraged to enter the academy by Australian university policies and government legislation but, once there, find that the promise of equal opportunity is often not kept. This article provides some insights into how university lecturers' normative expectations and practices can affect students' everyday experiences. Interviews with students with LD showed that they do not often receive support services, their "stories" are not believed, and they often feel that they do not "belong". PMID- 17915499 TI - Innovative programs for improvement in reading through cognitive enhancement: a remediation study of Canadian First Nations children. AB - Forty-five Grade 3 students from a reservation school in Western Canada were divided into two remedial groups and a no-risk control group. One remedial group was given a classroom-administered cognitive enhancement program (COGENT) throughout the school year. The second group received COGENT for the first half of the year followed by a pull-out cognitive-based reading enhancement program (PREP). Children were assessed across phonological awareness, rapid naming, reading, and cognitive ability at the beginning of the year, mid-term, and at the end of the school year. MANOVA results showed a significant interaction for reading measures, with students receiving classroom intervention over the school year making the greatest gains. Results are discussed in terms of group, remediation program, and individual participant improvements. PMID- 17915500 TI - Cognitive characteristics of children with mathematics learning disability (MLD) vary as a function of the cutoff criterion used to define MLD. AB - Researchers of mathematics learning disability (MLD) commonly use cutoff scores to determine which participants have MLD. Some researchers apply more restrictive cutoffs than others (e.g., performance below the 10th vs. below the 35th percentile). Different cutoffs may lead to groups of children that differ in their profile of math and related skills, including reading, visual-spatial, and working memory skills. The present study assesses the characteristics of children with MLD based on varying MLD definitions of math performance either below the 10th percentile (n = 22) or between the 11th and 25th percentile (n = 42) on the Test of Early Math Ability, second edition (TEMA-2). Initial starting levels and growth rates for math and related skills were examined in these two MLD groups relative to a comparison group (n = 146) whose TEMA-2 performance exceeded the 25th percentile. Between kindergarten and third grade, differences emerged in the starting level and growth rate, suggesting qualitative differences among the three groups. Despite some similarities, qualitative group differences were also observed in the profiles of math-related skills across groups. These results highlight differences in student characteristics based on the definition of MLD and illustrate the value of examining skill areas associated with math performance in addition to math performance itself. PMID- 17915501 TI - Polymerase chain reaction identification of three members of the Anopheles sundaicus (Diptera: Culicidae) complex, malaria vectors in Southeast Asia. AB - Anopheles sundaicus s.l., a major malaria vector taxon, occurs primarily along coastal areas and on islands in Southeast Asia. Our previous studies using cytochrome oxidase I, cytochrome-b, and internal transcribed spacer 2 markers discriminated three allopatric species: An. sundaicus s.s. in northern Borneo, An. epiroticus in Southeast Asia, and An. sundaicus E on Sumatra and Java, Indonesia. Morphological comparisons of three developmental stages did not reveal unique diagnostic characters that could reliably distinguish the three species. Therefore, we developed a multiplex polymerase chain reaction (PCR) assay based on two mitochondrial DNA markers to unambiguously identify them. This PCR was tested on 374 specimens from 24 different geographical populations, expanding our knowledge of the distribution of these species. PMID- 17915502 TI - Life cycles of seven ixodid tick species (Acari: Ixodidae) under standardized laboratory conditions. AB - Studies of transmission, maintenance, infectivity, virulence, and pathogenicity of tick-borne agents require the use of large numbers of live laboratory-raised ticks. Colonies of Ixodes scapularis Say, Ixodes pacificus Cooley & Kohls, Amblyomma americanum (L.), Dermacentor occidentalis Marx, Dermacentor variabilis (Say), Hemaphysalis leporispalustris (Packard), and Rhipicephalus sanguineus (Latrielle) have been maintained in our laboratory at the Centers for Disease Control and Prevention for five to 18 continuous generations. New Zealand White rabbits (Oryctolagus cuniculus) are used as hosts for all tick species and developmental stages. Between feedings, ticks are stored in environmental incubators at 22-24 degrees C and 90% RH with a day/night photoperiod of 16:8 (L:D) h. The duration of feeding, molting, preoviposition, and periods of postmolting development were recorded. Here, we describe the life cycles of these common North American tick species under standardized laboratory conditions. At 22-24 degrees C, the minimal time needed for each species to complete one life cycle was as follows: I. scapularis, 204-219 d; I. pacificus, 214-229 d; R. sanguineus, 162-177 d; H. leporispalustris, 209-224 d; D. variabilis, 176-191 d; D. occidentalis, 180-195 d; and A. americanum, 192-211 d. PMID- 17915503 TI - Effect of temperature on life history parameters of adult Culicoides sonorensis (Diptera: Ceratopogonidae) in relation to geographic origin and vectorial capacity for bluetongue virus. AB - The effect of temperature on survival, oviposition, gonotrophic development, and a life history factor of vectorial capacity were examined in adult Culicoides sonorensis (Wirth & Jones) (Diptera: Ceratopogonidae) that originated from two geographic locations. Flies originating from the United States (Colorado) had slightly reduced survival after a bloodmeal compared with wild flies collected in southern Alberta (AB), Canada. Survival of AB flies declined in a curvilinear manner with temperature, whereas survival of U.S. flies showed a linear response to temperature. The survivorship curve of the AB flies more closely followed a Weibull distribution than an exponential, indicating survival was age-dependent. Survivorship of the U.S. flies followed an exponential distribution. Females from both sources laid similar numbers of eggs throughout their life. The first eggs were laid by females from both sources at 31.9 degree-day (DD)9.3. Dissections of blood-fed flies reared at various temperatures indicated that flies from both sources were 90% gravid at 32 DD9.3. Relationships among temperature and life history components of vectorial capacity were similar among flies from the two sources and indicated that vectorial capacity would be approximately 1.8-2.6-fold greater in a southern U.S. climate compared with southwestern Canada due solely to the effects of temperature on the life history of C. sonorensis. Using life history estimates derived from Weibull model had little effect on estimating vectorial capacity, whereas using estimates derived from the exponential model slightly overestimated vectorial capacity. PMID- 17915504 TI - Effects of reduced deer density on the abundance of Ixodes scapularis (Acari: Ixodidae) and Lyme disease incidence in a northern New Jersey endemic area. AB - We monitored the abundance of Ixodes scapularis Say (Acari: Ixodidae) and the Lyme disease incidence rate after the incremental removal of white-tailed deer, Odocoileus virginianus Zimmermann, within a suburban residential area to determine whether there was a measurable decrease in the abundance of ticks due to deer removal and whether the reduction in ticks resulted in a reduction in the incidence rate within the human population. After three seasons, the estimated deer population was reduced by 46.7%, from the 2002 postfawning estimate of 2,899 deer (45.6 deer per km2) to a 2005 estimate of 1,540 deer (24.3 deer per km2). There was no apparent effect of the deer culling program on numbers of questing I. scapularis subadults in the culling areas, and the overall numbers of host seeking ticks in the culling areas seemed to increase in the second year of the program. The Lyme disease incidence rate generated by both passive and active surveillance systems showed no clear trend among years, and it did not seem to vary with declining deer density. Given the resources required to mount and maintain a community-based program of sufficient magnitude to effectively reduce vector tick density in ecologically open situations where there are few impediments to deer movement, it may be that deer reduction, although serving other community goals, is unlikely to be a primary means of tick control by itself. However, in concert with other tick control interventions, such programs may provide one aspect of a successful community effort to reduce the abundance of vector ticks. PMID- 17915505 TI - Survivorship of immature stages of Anopheles gambiae s.l. (Diptera: Culicidae) in natural habitats in western Kenya highlands. AB - We examined the survivorship of Anopheles gambiae s.l. Giles (Diptera: Culicidae) larvae and habitat productivity in three major habitat types in the western Kenya highlands. The age-specific distribution was determined for larvae and pupae, and survivorship curves were constructed. Larval-to-pupal survivorship was 6.8% in drainage ditches, 4.3% in cow hoofprints, and 1.8% in disused goldmines, respectively. High mortality rates were observed in all developmental stages. The estimated daily survival rate was highest in drainage ditches (0.74), followed by cow hoofprints (0.71), and it was lowest in disused goldmines (0.62). Productivity of emerging An. gambiae adults was generally low in these larval habitats (1.35, 1.55, and 1.84 mosquitoes per m2 per wk in drainage ditches, disused goldmines, and cow hoofprints, respectively). In total, seven families of larval mosquito predators were identified from the larval habitats, including Hydrophilidae, Dytiscidae, Corixidae, Nepidae, Notonectidae, Belostomatidae, and Cordulidae. Predator density in disused goldmines was significantly higher than that of other habitat types. Determination of the relative importance of predation, habitat stability and food contents on natural mosquito habitat productivity would help to design cost-effective vector control methods specifically targeted at the productive habitats. PMID- 17915506 TI - Stable fly population dynamics in eastern Nebraska in relation to climatic variables. AB - Stable flies, Stomoxys calcitrans (L.), are among the most economically important arthropod pests of livestock in North America. In this study, we monitored the seasonal dynamics of a stable fly population in eastern Nebraska for 5 yr. Models based upon temperature and precipitation were developed to determine the affects of these variables on population levels as well as to project population trends. Stable flies appear in eastern Nebraska in late March to early April, and they build to a peak population during the last week of June and first week of July. In most years, the population decreases in midsummer, and then it increases to a second peak in mid-September. Temperature 0 to 2 wk before collection and precipitation 3 to 6 wk before collection were the most important weather variables accounting for 63 and 11% of the variation, respectively. Temperature 7 wk before collection was also significant, accounting for 3% of the variation. Reduced precipitation levels explained the observed midsummer drop in the stable fly populations. Changes in stable fly population levels were positively correlated with precipitation 1 to 2 wk prior and temperature the week of the change. Population change was negatively correlated with precipitation 6-8 wk prior and temperature 6-15 wk prior. The addition of the previous weeks trap collections to the climate based model eliminated the significance of temperature 2 and 7 wk before collection. Temperature 0-1 wk before collection accounted for 60% of the variation, precipitation 3 to 6 wk prior 12% of the variation, and the previous weeks' trap collections accounted for 11% of the variation. Low temperatures during October through January were correlated with higher stable fly populations the following June and July. PMID- 17915507 TI - Temperature-dependent demography of Supella longipalpa (Blattodea: Blattellidae). AB - The demography of the brownbanded cockroach, Supella longipalpa (F.) (Blattodea: Blattellidae), was studied based on the age-stage, two-sex life table at 25, 29, and 33 degrees C. Females incubated at the three temperatures produced 11.8, 14.6, and 12.8 oothecae per female, respectively. The life expectancy for a newborn was 157.2, 207.7, and 147.9 d, respectively. The intrinsic rate of increase at these temperatures was 0.0161, 0.0306, and 0.0398 d(-1), respectively. The net reproductive rate was 35.3, 100.9, and 87.2 offspring, respectively. The mean generation time was 222.1, 151.1, and 112.5 d, respectively. In the absence of other limiting factors, our results indicate that populations of S. longipalpa would be expected to establish and increase if introduced into environments where temperature was within 25 and 33 degrees C. PMID- 17915508 TI - Host odor synergizes attraction of virgin female Lutzomyia longipalpis (Diptera: Psychodidae). AB - The sand fly Lutzomyia longipalpis Lutz & Neiva (Diptera: Psychodidae) is the principle vector of Leishmania chagasi/infantum Cunha and Chagas, the causative agent of visceral leishmaniasis. The disease is transmitted by blood-feeding females, which seek aggregations of males above potential hosts both to mate and blood-feed. Pheromones produced by male sand flies could potentially be used as lures in L. longipalpis control programs. We investigated whether attraction of male and female sand flies to pheromone could be increased by addition of host odor. Pheromone was attractive to females in the absence of host odor, although a 10-fold increase in concentration did not increase numbers attracted or reduce the proportion of flies not responding during trials. Odors from Syrian hamsters, Mesocricetus auratus, were more attractive to females than air when presented without sex pheromone, but this effect was masked by hexane, suggesting attraction to host odor alone may be relatively weak. Addition of hamster odor both increased the number of virgin female L. longipalpis attracted to sex pheromone (relative to a solvent control) and reduced the number of nonresponders, indicating that host odor may have both a synergistic and activating effect. Male sand flies were not attracted to pheromone with or without host odor, although addition of pheromone did counteract an apparent avoidance of host odor combined with a hexane control. These results indicate L. longipalpis pheromones function primarily to attract females and that their efficacy as lures may be increased through addition of host odor, or by deploying traps in the vicinity of host animals. PMID- 17915509 TI - Population structuring of Glossina palpalis gambiensis (Diptera: Glossinidae) according to landscape fragmentation in the Mouhoun river, Burkina Faso. AB - The impact of landscape fragmentation due to human and climatic mediated factors on the structure of a population of Glossina palpalis gambiensis Vanderplank (Diptera: Glossinidae) was investigated in the Mouhoun river basin, Burkina Faso. Allele frequencies at five microsatellite loci, and metric properties based on 11 wing landmarks, were compared between four populations. The populations originated from the Mouhoun river and one of its tributaries. The average distance between samples was 72 km with the two most widely spaced populations being 216 km apart. The sampling points traversed an ecological cline in terms of rainfall and riverine forest ecotype, along a river enlarging from downstream to upstream and oriented south to north. Microsatellite DNA comparison demonstrated structuring between the populations, but not complete isolation, with an overall Fst = 0.012 (P < 0.001). Wing geometry revealed significant centroid size and shape differences between populations, especially between the two most distant populations. There was no significant correlation between gene flow and geographic distance at this scale, but there was a positive correlation in females between metric distances (wing shape differences) and geographic distances that might be attributed to the cline of environmental conditions. The impact of the fragmentation of riparian landscapes on tsetse population structure is discussed in the context of control campaigns currently promoted by Pan African Tsetse and Trypanosomosis Eradication Campaign. PMID- 17915510 TI - Detection of pyrethroid resistance gene in head lice in schoolchildren from Bobigny, France. AB - The head louse, Pediculus humanus capitis (De Geer), is an hematophagous ectoparasite that affects mainly children. Resistance to insecticides belonging to pyrethroids and other pediculicides, such as malathion, is responsible for frequently reported treatment failures. Recent studies showed that a M815I-T929I L932F kdr-like mutation in the voltage-gated sodium channel alpha-subunit gene was associated with permethrin resistance in head lice from several countries worldwide. We searched for the presence ofpyrethroid resistance gene in head lice populations obtained in schoolchildren in an urban area of France. All the 15 primary schools of Bobigny, a city located 3 km north of Paris, were selected to participate. Of 3,493 children enrolled, 3,345 (95.8%) children were screened for head lice by using fine-toothed antilouse combs. Live head lice were detected in 112 (3.3%) of children screened. A subsample of 90 lice was processed for DNA study. The amplification of a 332-bp portion of the kdr-like gene spanning the codon 929 was performed, and polymerase chain reaction products were submitted to the restriction enzyme SspI. Twenty of these lice (22.2%) were homozygous susceptible, 33 (36.7%) were homozygous resistant, and 37 (41.1%) were heterozygotes. Globally, the frequency of the T929I mutation was 0.57. The prevalence of pediculosis in schoolchildren of Bobigny seemed relatively low in comparison with findings of other European studies. The presence of the T929I mutation associated with permethrin resistance probably reflected the frequent local use of this insecticide. Further studies are now required to evaluate the prevalence of the kdr-like mutant allele in head lice in French schools. PMID- 17915511 TI - Ovicidal activity of entomopathogenic hyphomycetes on Aedes aegypti (Diptera: Culicidae) under laboratory conditions. AB - The ovicidal activity of 21 hyphomycete fungi species against Aedes aegypti (L.) (Diptera: Culicidae) was tested. Fungi with ovicidal activity developed on high numbers of eggs (> or =70%) during 25 d of exposure. A clear ovicidal activity with low values of hatch (1.3-40%) was observed after 25 d of incubation with Isaria farinosa (Holm: Fries) Fries, Paecilomyces carneus (Duche & Heim) Brown & Smith, Paecilomyces marquandii (Massee) Hughes, Isaria fumosorosea (Wize), Metarhizium anisopliae (Metschnikoff) Sorokin, Penicillium sp., Paecilomyces lilacinus (Thom) Samson, Beauveria bassiana (Balsamo) Vuillemin, and Evlachovaea kintrischica Borisov & Tarasov. More than 63% of eggs hatched after 25-d exposures to 11 other fungi species deemed as ineffective. These are the first results to show the effects of entomopathogenic fungi against eggs of Ae. aegypti, and they suggest their potential as control agents of this vector. PMID- 17915512 TI - Characterization of insensitive acetylcholinesterase (ace-1R) in Anopheles gambiae (Diptera: Culicidae): resistance levels and dominance. AB - Characterization of insecticide resistance provides data on the evolutionary processes involved in the adaptation of insects to environmental changes. Studying the dominance status and resistance level represents a great interest, in terms of understanding resistance evolution in the field to eventually adapt vector control. Resistance and dominance levels conferred by the G119S mutation of acetylcholinesterase (ace-1R) of the mosquito Anopheles gambiae s.s. (Diptera: Culicidae) were studied for various insecticides belonging to different classes, using strains sharing the same genetic background. Our survey shows that the homozygote resistant strain AcerKis displayed a very high resistance level to various carbamates (range 3,000- to 5,000-fold) compared with that of various organophosphates (range 12- to 30-fold). Furthermore, the dominance status varied between semi-recessivity with fenitrothion and chlorpyrifos methyl insecticides to semidominance with temephos, carbosulfan, and propoxur. These results indicate that this resistance mechanism could spread rapidly in the field and then compromise the use of organophosphate and carbamate compounds in public health. This study underlines the necessity to monitor the ace-1R mutation in natural populations before planning and implementing malaria control programs based on the use of these insecticides. PMID- 17915513 TI - Impact of inorganic pollutants perchlorate and hexavalent chromium on efficacy of Bacillus sphaericus and Bacillus thuringiensis subsp. israelensis against Culex quinquefasciatus (Diptera: Culicidae). AB - The effects of two widespread environmental pollutants, perchlorate and hexavalent chromium, were assessed on the efficacy of Bacillus thuringiensis subsp. israelensis (Bti) and Bacillus sphaericus (Bsph) against fourth instars of Culex quinquefasciatus Say (Diptera: Culicidae) in 24-h laboratory bioassays. Although 250 mg/liter perchlorate, a level somewhat higher than would be considered ecologically relevant, did not affect the control provided by either larvicide, presence of 1.04 mg/liter hexavalent chromium, an ecologically relevant concentration, increased the efficacy of both Bti and Bsph by 21 and 80%, respectively. In the presence of hexavalent chromium, improved suppression could be expected from Bacillus applications at the current label rates. However, because hexavalent chromium has been shown to affect many taxa, we propose that the potential exists for increased susceptibility of nontarget organisms to Bacillus products in polluted habitats. PMID- 17915514 TI - Acaricide rotation strategy for managing resistance in the tick Rhipicephalus (Boophilus) microplus (Acarina: Ixodidae): laboratory experiment with a field strain from Costa Rica. AB - During the past two decades, resistance to pyrethroids within the cattle tick genus Boophilus has caused tick control problems in various tropical countries, mainly in Latin America, southern Africa, Australia, and New Caledonia. A Rhipicephalus (Boophilus) microplus (Canestrini) strain from Costa Rica, exhibiting resistance to the pyrethroid deltamethrin but only a very low resistance to organophosphates (OP) was kept under selection pressure for 9 to 11 generations by using deltamethrin or coumaphos (OP), either exclusively or in rotation. The objective of this acaricide rotation was to examine the possibility of delaying or reducing the full emergence of pyrethroid resistance. In the substrain selected with deltamethrin at the LD50 concentration, resistance to deltamethrin was measured after five generations (resistance factor [RF] = 9.2) and very high resistance after 11 generations (RF = 756), compared with the starting field strain from Costa Rica. In the substrain selected with deltamethrin then coumaphos in rotation, resistance to deltamethrin was variable from one generation to the next (RF = 1-4.2), but no high, stable resistance developed. After 10 generations of rotation, the deltamethrin RF was 1.6. In the substrains selected continuously with coumaphos or coumaphos and deltamethrin in rotation, no consistent change in resistance to coumaphos was observed. Rotation of deltamethrin with coumaphos seems to delay the development of strong resistance to deltamethrin in a population that had initially a low level of deltamethrin resistance. PMID- 17915515 TI - Insecticide resistance in potential vector mosquitoes for West Nile virus in Japan. AB - Culex pipiens complex is the significant vector mosquito of West Nile virus. To take stock of the current situation of insecticide susceptibilities and design an ideal mosquito control strategy, we collected Culex pipiens pallens Coquillet, Culex pipiens form molestus Forskal, and Culex quinquefasciatus Say from fields in Japan and conducted bioassays for five larvicides (fenitrothion, temephos, etofenprox, diflubenzuron, and pyriproxyfen) by using a larval dipping method. Among five insecticides tested, obvious reduced susceptibilities were observed for etofenprox, which is the only pyrethroid compound registered as a larvicide in Japan. Twenty-two of 56 colonies exhibited a >10% survival rate at the etofenprox concentration of 5.7 microg/ml, which is a 10 times higher concentration of the working solution. The LC50 of a colony collected from Fukuoka prefecture for etofenprox exceeded 60 microg/ml (resistance ratio >2,307), and this colony also exhibited cross-resistance to other pyrethroids, permethrin (299-fold) and phenothrin (1,200-fold). The insect growth regulators diflubenzuron and pyriproxyfen were found to be sufficiently effective enough to control Culex larvae present, but decreased sensitivities to these insecticides were slightly detected in some colonies of Cx. p. form molestus collected from urban areas. Several etofenprox-resistant colonies of Cx. p. form molestus exhibited simultaneously decreased susceptibilities to other insecticides, including temephos, diflubenzuron, and pyriproxyfen. PMID- 17915516 TI - Integrated use of 4-Poster passive topical treatment devices for deer, targeted acaricide applications, and Maxforce TMS bait boxes to rapidly suppress populations of Ixodes scapularis (Acari: Ixodidae) in a residential landscape. AB - In fall 2003, we began testing an integrated control strategy to rapidly achieve and sustain reduced numbers of Ixodes scapularis Say (Acari: Ixodidae) in a residential area. We combined two host-targeted technologies in conjunction with single, barrier acaricide applications to sequentially attack each postembryonic life stage of the tick. Granular deltamethrin applied to the lawn-forest interface of participant properties resulted in 100% control of host-seeking nymphs. Nymphal and larval tick burdens on targeted small mammal hosts at treated properties were reduced by 92.7 and 95.4%, respectively, after the first year (2004) of combined interventions. Over the same period, populations of host seeking nymphs, larvae, and adults were reduced by 58.5, 24.8, and 77.8%, respectively. After interventions in 2005, tick burdens on small mammals were maintained at similar levels, whereas control of host-seeking nymphs, larvae, and adults increased to 94.3, 90.6, and 87.3%, respectively. Prospects for widespread use of these technologies to protect the public's health are discussed. PMID- 17915517 TI - Detection and isolation of exotic Newcastle disease virus from field-collected flies. AB - Flies were collected by sweep net from the vicinity of two small groups of "backyard" poultry (10-20 chickens per group) that had been identified as infected with exotic Newcastle disease virus (family Paramyxoviridae, genus avulavirus, ENDV) in Los Angeles County, CA, during the 2002-2003 END outbreak. Collected flies were subdivided into pools and homogenized in brain-heart infusion broth with antibiotics. The separated supernatant was tested for the presence of ENDV by inoculation into embryonated chicken eggs. Exotic Newcastle disease virus was isolated from pools of Phaenicia cuprina (Wiedemann), Fannia canicularis (L.), and Musca domestica L., and it was identified by hemagglutination inhibition with Newcastle disease virus antiserum. Viral concentration in positive pools was low (<1 egg infectious dose50 per fly). Isolated virus demonstrated identical monoclonal antibody binding profiles as well as 99% sequence homology in the 635-bp fusion gene sequence compared with ENDV recovered from infected commercial egg layer poultry during the 2002 outbreak. PMID- 17915518 TI - Expectoration of Flaviviruses during sugar feeding by mosquitoes (Diptera: Culicidae). AB - Biological transmission of arboviruses to a vertebrate host occurs when virions are expelled along with saliva during blood feeding by a hematophagous arthropod. We undertook experiments to determine whether mosquitoes expectorate flaviviruses in their saliva while sugar feeding. Batches of Culex annulirostris Skuse and Culex gelidus Theobald (Diptera: Culicidae) were orally infected with Japanese encephalitis (family Flaviviridae, genus Flavivirus, JEV), Kunjin (family Flaviviridae, genus Flavivirus, KUNV; a subtype of West Nile virus), and Murray Valley encephalitis (family Flaviviridae, genus Flavivirus, MVEV) viruses. After a 7-d extrinsic incubation, these mosquitoes were offered sucrose meals via cotton pledgets, which were removed daily and processed for viral RNA by using real-time TaqMan reverse transcriptase-polymerase chain reaction (RT-PCR) assays. JEV, MVEV, and KUNV RNA was detected in all pledgets removed from batches of Cx. gelidus on days 7-14 postexposure. In contrast, detection rates were variable for Cx. annulirostris, with KUNV detected in 0.3 M sucrose pledgets on all days postexposure, and JEV and MVEV detected on 57 and 50% of days postexposure, respectively. Higher concentrations of sucrose in the pledget did not increase virus detection rates. When individual JEV-infected Cx. gelidus were exposed to the sucrose pledget, 73% of mosquitoes expectorated virus with titers that were detectable by TaqMan RT-PCR. These results clearly show that flaviviruses are expectorated by infected mosquitoes during the process of sugar feeding on artificial pledgets. Potential applications of the method for arboviral bioassays and field surveillance are discussed. PMID- 17915519 TI - Characterization of Aedes aegypti (Diptera: Culcidae) production sites in urban Nicaragua. AB - To characterize the production patterns of the dengue virus vector Aedes aegypti (L.) (Diptera: Culcidae), pupal surveys were conducted in selected neighborhoods of two major cities in Nicaragua. In Leon, 833 houses were visited in July and September 2003, corresponding to the beginning and middle of the dengue season; in Managua, 1,365 homes were visited in July 2003. In total, 7,607 containers were characterized, of which 11% were positive for Ae. aegypti larvae and 4% for pupae. In addition to barrels, potted plants and superficial water on tarps and in puddles were identified as highly productive sites. Univariate and multivariate analysis revealed frequency of container use, use of a lid, and rainwater filling as key variables affecting pupal positivity. Importantly, this survey demonstrated the risk associated with the presence of lids, the limited temporal efficacy of temephos, and the lack of association of water availability with risky water storage practices. Finally, we introduce the concept of an efficiency value and an accompanying graphical display system that can facilitate development of targeted pupal control strategies. These data underscore the importance of entomological surveillance of pupal productivity to gather information from which to derive streamlined, efficient, and effective vector control measures to reduce the density of Aedes mosquito larvae and pupae and thus the risk for dengue. PMID- 17915520 TI - Landscape factors influencing the spatial distribution and abundance of mosquito vector Culex quinquefasciatus (Diptera: Culicidae) in a mixed residential agricultural community in Hawai'i. AB - Mosquito-borne avian diseases, principally avian malaria (Plasmodium relictum Grassi and Feletti) and avian pox (Avipoxvirus sp.) have been implicated as the key limiting factor associated with recent declines of endemic avifauna in the Hawaiian Island archipelago. We present data on the relative abundance, infection status, and spatial distribution of the primary mosquito vector Culex quinquefasciatus Say (Diptera: Culicidae) across a mixed, residential agricultural community adjacent to Hawai'i Volcanoes National Park on Hawai'i Island. We modeled the effect of agriculture and forest fragmentation in determining relative abundance of adult Cx. quinquefasciatus in Volcano Village, and we implement our statistical model in a geographic information system to generate a probability of mosquito capture prediction surface for the study area. Our model was based on biweekly captures of adult mosquitoes from 20 locations within Volcano Village from October 2001 to April 2003. We used mixed effects logistic regression to model the probability of capturing a mosquito, and we developed a set of 17 competing models a priori to specifically evaluate the effect of agriculture and fragmentation (i.e., residential landscapes) at two spatial scales. In total, 2,126 mosquitoes were captured in CO2-baited traps with an average probability of 0.27 (SE = 0.10) of capturing one or more mosquitoes per trap night. Twelve percent of mosquitoes captured were infected with P. relictum. Our data indicate that agricultural lands and forest fragmentation significantly increase the probability of mosquito capture. The prediction surface identified areas along the Hawai'i Volcanoes National Park boundary that may have high relative abundance of the vector. Our data document the potential of avian malaria transmission in residential-agricultural landscapes and support the need for vector management that extends beyond reserve boundaries and considers a reserve's spatial position in a highly heterogeneous landscape. PMID- 17915522 TI - Comparative performance and complementarity of four sampling methods and arthropod preference tests from human and porcine remains at the Forensic Anthropology Center in Knoxville, Tennessee. AB - Comparative performance and complementarity tests of four arthropod sampling methods (aerial netting, hand collection, pitfall traps, and sticky traps), used by forensic entomologists in death investigations, training workshops, and research trials, were conducted from simultaneously placed human and porcine subjects inside the Forensic Anthropology Center at the University of Tennessee, Knoxville, TN. A secondary aim investigated the widely held claim that pig carcasses are reliable surrogates for human corpses. Over a 35-d period in summer 1989, >72,000 invertebrates from three subjects (one human, two pigs) were sampled of which 93% were members of the forensically important (FI) fauna. Performance tests revealed that hand collections, when performed by an experienced forensic entomologist, consistently yielded the largest fraction of FI arthropods from the total invertebrate catch, followed by aerial netting, sticky traps, and pitfall traps, regardless of subject. Pitfall traps and hand collections were broadly effective at sampling both fly and beetle populations, whereas aerial netting and sticky traps mostly targeted flies. The best two method combination, based on the highest combined catches of FI taxa, were hand collections and pitfall traps, regardless of subject. Between-subject comparisons revealed negligible preference by FI arthropods for human over pig remains. Insofar as our limited comparisons allow with only three study subjects, these results validated the concept of transferability of "best practices" from one subject to another and confirmed the claim that pig carcasses (of 23-27-kg starting mass) can substitute for human corpses in research and training programs, at least for summer-exposed and unconcealed remains in the first 5 wk postmortem. PMID- 17915521 TI - Molecular identification of bloodmeal source in Ixodes ricinus ticks using 12S rDNA as a genetic marker. AB - We developed an efficient molecular method for the identification of the bloodmeal sources in the tick Ixodes ricinus (L.), the European vector of the agents of Lyme borreliosis and tick-borne encephalitis. An approximately 145-bp orthologous fragment of the vertebrate mitochondrial 12S rDNA was used as a molecular marker to discriminate host vertebrate species. The method consists of a single run polymerase chain reaction amplification of the 12S rDNA molecular marker by using nondegenerate primers followed by a reverse line blot hybridization assay by using specific oligonucleotide probes. The palette of probes allowed us to distinguish major groups of host vertebrates (e.g., mammals, small rodents, artiodactyls, birds, lizards) and to identify the bloodmeal sources at the genus or species level. External primers were designed and used to sequence the 12S rDNA molecular marker of a broad range of known or potential host vertebrate species (n = 60), including mammal (n = 28), bird (n = 31), and reptile (n = 1) species. The use of this technique coupled with known methods for identification of tick-borne pathogens (e.g., Borrelia burgdorferi sensu lato) allowed us to determine the source of infective bloodmeal and to identify reservoir species. The present method was successfully used to identify the source of bloodmeals in all feeding I. ricinus ticks and in half of questing field-collected I. ricinus ticks. Moreover, the bloodmeal source was identified in 65% of ticks infected with B. burgdorferi sensu lato. Further development of this technique may be envisaged for the detection of other vector-borne pathogens and their reservoir hosts. PMID- 17915523 TI - Characterization of permethrin-resistant Boophilus microplus (Acari: Ixodidae) collected from the state of Coahuila, Mexico. AB - Southern cattle ticks, Boophilus microplus (Canestrini) (Acari: Ixodidae), collected in Coahuila, Mexico, were determined to be resistant to permethrin. Discriminating concentration (DC) tests at the LC99 and 2X the LC99 of susceptible ticks produced 0 and 0.5% mortality, respectively, for permethrin. However, measured mortalities for coumaphos and amitraz acaricides were within the expected ranges. Polymerase chain reaction analysis of this strain detected a 99% frequency of homozygous resistant individuals in this strain. PMID- 17915524 TI - Seasonal variation in nymphal blacklegged tick abundance in southern New England forests. AB - In the northeastern United States, risk of human exposure to tick transmitted disease is primarily a function of the abundance of the blacklegged tick, Ixodes scapularis Say. We assessed seasonal variability in the abundance of nymphal stage I. scapularis over 13 yr, collected from several forested areas throughout Rhode Island. Specifically, we examined intraseasonal differences by using two temporally distinct tick collections made during the peak nymphal tick season. Intraseasonal factors significantly impacted tick abundance, with the June tick rate (mean = 40.42, SD = 14.79) significantly more abundant than the July tick rate (mean = 27.64, SD = 15.47). The greater variability in July (coefficient of variation: June, 36.61%; July, 55.95%) lead us to conclude June tick rates are relatively stable from year to year, whereas July tick rates contribute more to intraseasonal and yearly variation. PMID- 17915525 TI - Bothriocroton concolor (Acari: Ixodidae) on the Kangaroo Island kangaroo: a new host-parasite relationship. AB - In 2006, we examined Kangaroo Island kangaroos, Macropusfuliginosusfuliginosus, for ticks. We collected three tick species: Ixodes hirsti Hassall, Hemaphysalis bancrofti Nuttall & Warburton, and Bothriocroton concolor (Neumann). Surprisingly, the specimens included eight females and one nymph of B. concolor, which had previously been considered strictly host specific to the echidna (Tachyglossus sp.). This is the first record of B. concolor on the Kangaroo Island kangaroo. PMID- 17915526 TI - Eliminating adverse drug events at Ascension Health. AB - Improvement in ADE rate did not occur at either hospital with any one particular intervention, or even after several conventional interventions had time to take effect. Instead, instituting many simultaneous system changes, proactively assessing risk, and expanding the scope of intervention were each essential to sustaining the described reductions in harm. The next step toward eliminating ADEs requires simultaneous communication across systems in a way that is manageable, approachable, and adaptable, and that supports the elements of change. A system of mutually informed processes--from medication selection and entry through preparation and dispensing, administration, monitoring, and reconciliation--should result in safe, patient-centered, reliable, and efficient medication use. In March 2007, Ascension Health, drawing on the work described in this article, began developing such a system. PMID- 17915527 TI - Preventing harm from high-alert medications. AB - BACKGROUND: One of the 12 interventions that the Institute for Healthcare Improvement (IHI) recommends for its 5 Million Lives Campaign is "Prevent Harm from High-Alert Medications . . . starting with a focus on anticoagulants, sedatives, narcotics, and insulin." EXECUTING SYSTEM-LEVEL CHANGES: Three essential elements are needed to execute system-level changes in an organization: will, ideas, and execution. Will is developed by examining the status quo in an organization and agreeing that it is no longer acceptable. Ideas-changes that will make the system safer-can be found in the literature and in the experience of other hospitals and are the basis for the recommended general interventions to reduce errors and harm associated with high-alert medications. Execution, the process of making those changes real, requires commitment from senior leaders and clinical leaders, along with the organizational capacity to improve. The steps in the medication system are so interrelated that a change in one area will affect others' ability to complete their work. In addition, senior leadership and clinical leadership must visibly support the effort, connecting the reduction in high-alert medication-related harm to the overall hospital goal of harm reduction is essential. CONCLUSION: The campaign's goal is to achieve a 50% reduction in harm related to high-alert medications. Employing strategies such as standardization and simplification will provide the foundation for improved medication safety. PMID- 17915528 TI - Case study: Reducing narcotic oversedation across an integrated health system. PMID- 17915529 TI - Using the Communication and Teamwork Skills (CATS) Assessment to measure health care team performance. AB - BACKGROUND: Patient safety administrators, educators, and researchers are striving to understand how best to monitor and improve team skills and determine what approaches to monitoring best suit their organizations. A behavior-based tool, based on principles of crisis resource management (CRM) in nonmedical industries, was developed to quantitatively assess communication and team skills of health care providers in a variety of real and simulated clinical settings. THE CATS ASSESSMENT: The Communication and Teamwork Skills (CATS) Assessment has been developed through rapid-cycle improvement and piloted through observation of videotaped simulated clinical scenarios, realtime surgical procedures, and multidisciplinary rounds. Specific behavior markers are clustered into four categories-coordination, cooperation, situational awareness, and communication. Teams are scored in terms of the occurrence and quality of the behaviors. The CATS Assessment results enable clinicians to view a spectrum of scores-from the overall score for the categories to specific behaviors. CONCLUSION: The CATS Assessment tool requires statistical validation and further study to determine if it reliably quantifies health care team performance. The patient safety community is invited to use and improve behavior-based observation measures to better evaluate their training programs, continue to research and improve observation methodology, and provide quantifiable, objective feedback to their clinicians and organizations. PMID- 17915530 TI - Developing quality measures for sepsis care in the ICU. AB - BACKGROUND: Sepsis is associated with increased morbidity, mortality, and costs of care. Although several therapies improve outcomes in patients with sepsis, rigorously developed measures to evaluate quality of sepsis care in the intensive care unit (ICU) are lacking. METHODS: To select an initial set of candidate measures, in 2003-2004 an interdisciplinary panel reviewed the literature and used a modified nominal group technique to identify interventions that improve outcomes of patients with sepsis in the ICU. Design specifications or explicit definitions for each candidate measure were developed. RESULTS: Ten potential measures were identified: vancomycin administration, time to vancomycin initiation, broad-spectrum antibiotic administration, time to broad-spectrum antibiotic initiation, blood culture collection, steroid administration, corticotropin stimulation test administration, activated protein C eligibility assessment, activated protein C administration, and vancomycin discontinuation. DISCUSSION: The identification of potential measures of quality of care for patients with sepsis can help caregivers to focus on evidence-based interventions that improve mortality and to evaluate their current performance. Further work is needed to evaluate the feasibility and validity of the measures. PMID- 17915531 TI - Using an automated risk assessment report to identify patients at risk for clinical deterioration. PMID- 17915532 TI - The impact of abbreviations on patient safety. PMID- 17915533 TI - [Expression of regulatory and tissue-specific genes controlling regenerative potencies of the eye tissues in vertebrates]. AB - Comparative analysis of the early transformations of differentiated cells of the pigment epithelium, ciliary fold epithelium, and Muller glia in the eye of lower vertebrates and mammals during retina regeneration and cultivation was performed for the first time. Dedifferentiation and proliferation of cells and formation of progenitor multipotent cells, which are a source of retina regeneration in adult newts, were characterized using cell, molecular, and genetic markers. Neurospheres were formed during cultivation of the differentiated cells, in which progenitor multipotent cells were found that transformed into neurons of retina and brain and into glial cells. Comparative analysis of changes in the pigment epithelium cells during retina regeneration and during cultivation of differentiated cells of the pigment and ciliary epithelia and Muller glia suggests similar cell transformations at the early stages of transdifferentiation. PMID- 17915534 TI - [Effect of local microapplication of serotoninergic drugs on membrane currents of Paracentrotus lividus early embryos]. AB - It was shown that local application of agonists of the 3rd type receptors SR57277A and quipazine into the interblastomere cleft of Paracentrotus lividus embryos evoked specific membrane currents. At the same time, ligands of 5-HT3 receptors specifically affected the cleavage patterns of half-embryos, i.e., imitated or avoided the interblastomere signal. In the view of the data obtained, we discuss a more precise concept of protosynapse, where the distribution of membrane serotonin receptors is restricted to the period of blastomere formation during cleavage and localized in the area of interblastomere contact. PMID- 17915535 TI - [Preneural transmitters as regulators of embryogenesis. Current state of the problem]. AB - Our knowledge about the preneural neurotransmitter systems and their functions were based on the old pharmacological and biochemical data that have recently been confirmed and substantially supplemented. Specific components of the preneural serotoninergic and endocannabinoid systems were identified in developing echinoderm embryos using immunocytochemistry, Western immunoelectroblotting, and HPLC-mass spectroscopy. These data were corroborated by the results of pharmacological experiments: it was found that some ligands of serotonin receptors, as well as the agonist of cannabinoid receptors anandamide induced the appearance of abnormal embryonic phenotypes, whose expression depended on the ligand-teratogen concentration. Their appearance was prevented, correspondingly, by serotonin and its lipophilic (or hydrophilic) analogs and antagonists of cannabinoid (CB1/CB2)-receptors. PMID- 17915536 TI - [Biogenesis of plant fibers]. AB - The fiber (in terms of plant biology) is an individual cells characterized by spindle shape, length of up to several centimeters, well developed cell wall, and mechanical function. The review summarizes different, sometimes contradictory view points about duration, segregation and mechanisms of realization of individual stages of fiber biogenesis. Initiation and coordinated and intrusive growth are considered, as well as formation of secondary cell wall, including its gelatinous layers, and senescence. Biogenesis of fibers ontogenetically related to various tissues has been analyzed and the data about marker stage-specific characters of these cells. The data summarized in this review willow not only deeper understanding the development of cells with such unique characters, but also interpret the growth mechanisms for much more cell types, in which it is more difficult to identify individual stages of biogenesis than in the sclerenchyme fibers. PMID- 17915537 TI - [Interpopulation comparison of variations in gastrulation of Rana arvalis and Rana ridibunda occurring under the conditions of anthropogenic contamination of water bodies]. AB - The results of morphometric analysis of gastrulation in city populations of Rana arvalis and Rana ridibunda suggest that under the conditions of anthropogenic contamination of water bodies, the early development is more variable, if we compare less variable sizes of their eggs in the city and suburb populations. The variability of gastrulation appears to depend on the degree of contamination of spawning water bodies and is accompanied by increased correlation between the forming structures. This latter may be determined by two causes: (1) increased number of interdependent features, which leads to coordinated differentiation of structures and regulation of variability, and (2) increased coefficients of correlation within a small group of features, which leads to the formation of structures characteristic for the subsequent developmental stages, including the appearance of heterochronies. PMID- 17915538 TI - [Meiosis in the Siberian fir during microsporogenesis in conditions of arboretum]. AB - Meiosis was studied in the Siberian fir (Abies sibirica Ledeb.) at the arboretum of the Sukachev Institute of Forest. Specific features of meiosis in planted trees have been described. Both general and specific types of irregularities have been identified. The range of irregularities under the arboretum conditions was much wider than in natural ecosystems. PMID- 17915539 TI - Lean manufacturing comes to China: a case study of its impact on workplace health and safety. AB - Lean manufacturing, which establishes small production "cells," or teams of workers, who complete an entire product from raw material processing through final assembly and shipment, increases health and safety hazards by mixing previously separated exposures to various chemicals (with possible additive and cumulative effects) and noise. The intensification of work leads to greater ergonomic and stress-related adverse health effects, as well as increased safety hazards. The standard industrial hygiene approach of anticipation, recognition, evaluation, and hazard control is applicable to lean operations. A focus on worker participation in identifying and solving problems is critical for reducing negative impacts. A key to worker safety in lean production operations is the development of informed, empowered, and active workers with the knowledge, skills, and opportunity to act in the workplace to eliminate or reduce hazards. PMID- 17915540 TI - Lymphohematopoietic malignancies and oil exploitation in Koprivnica-Krizevci County, Croatia. AB - To determine lymphohematopoietic malignancy incidence rates between populations living near oil-gas fields (two regions) compared with those living in areas free from oil-gas exploitation (one region) in Koprivnica-Krizevci County, Croatia, 803 new cases of lymphohematopoietic malignancies were studied for two time periods: 1971-1980 (prior to oil-gas exploitation) and 1981-2000 (during oil-gas exploitation). The results showed considerable geographic differences in the incidences of chronic myeloid leukemia and multiple myeloma. The authors conclude that proximity to oil or gas fields represents an increased population risk of developing lymphohematopoietic cancers. PMID- 17915541 TI - A systematic review of the job-stress intervention evaluation literature, 1990 2005. AB - Ninety reports of systematic evaluations of job-stress interventions were rated in terms of the degree of systems approach used. A high rating was defined as both organizationally and individually focused, versus moderate (organizational only), and low (individual only). Studies using high-rated approaches represent a growing proportion of the job-stress intervention evaluation literature. Individual-focused, low-rated approaches are effective at the individual level, favorably affecting individual-level outcomes, but tend not to have favorable impacts at the organizational level. Organizationally-focused high- and moderate rated approaches are beneficial at both individual and organizational levels. Further measures are needed to foster the dissemination and implementation of systems approaches to examining interventions for job stress. PMID- 17915542 TI - Reducing pesticide exposure and associated neurotoxic burden in an Ecuadorian small farm population. AB - The contribution of community-based interventions, including farmer field schools (FFSs) in integrated pest management (IPM), to reducing pesticide exposures and associated neurotoxic burden among small-farm families in Ecuador was assessed in three Andean farming communities in a co-design of targeted action-research. Baseline questionnaire surveys elicited pesticide-related knowledge, practices, and exposure and neurobehavioral assessments were done using an adapted WHO battery. Pesticide applications on plots farmed by FFS versus non-FFS participants were compared. A year later, repeated surveys of participating households (n = 29) and neurobehavioral testing of individuals (n = 63) permitted comparisons of pre- and post-intervention values. The FFS graduates applied pesticides on their plots less frequently (p = 0.171). FFS households had increased pesticide-related knowledge of labels and exposure risk factors (both p < 0.004), better pesticide-handling practices (p < 0.01), and less skin exposure (p < 0.01). Neurobehavioural status had improved, particularly digit span and visuo-spatial function, resulting in overall z-score increases. Thus, community interventions reduced pesticide use, reported skin exposure, and neurotoxic burden among smallholder farm families. PMID- 17915544 TI - Saving the rainforest through health care: medicine as conservation in Borneo. AB - This article gives an overview of rainforest conservation as it relates to human health and describes the context, design, and implementation of the Kelay Conservation Health Program (KCHP). The KCHP is a health program for indigenous people living in a critical area of orangutan rainforest habitat in Indonesian Borneo also developed to aid conservation efforts there. Program design included consideration of both health and conservation goals, participatory planning in collaboration with the government health system, a focus on community managed health, capacity building, and adaptive management. After two years the program had, at relatively low cost, already had positive impacts on both human health (e.g., child immunization rates) and conservation (e.g., local forest protection measures, attitudes of villagers and government officials towards the implementing conservation agency). PMID- 17915543 TI - Occupational accidents and injuries in Thailand. AB - The Workers' Compensation Fund (WCF) represents only a limited fraction of work related injuries in the Thai workforce. This cross-sectional study examined 258,986 records from the Thai National Injury Surveillance (NIS) system collected during 2001-2004, focusing on 17,538 injuries coded as work-related. NIS records provided information generally not represented in WCF statistics, such as construction and agriculture. The reported mechanisms in 129 work-related fatalities were particularly informative, including electrical current (27%), transport accidents (20%), and falls (15%). Mortality in transport accidents was dramatically higher when seat belts or motorcycle helmets were not used, whether work-related or not. The findings emphasize the need to use multiple sources of information for a complete picture of work-related injuries in Thailand, and possibly in other countries. The mechanisms of fatal injury indicate areas where focused efforts are warranted. PMID- 17915545 TI - IBM, Elsevier Science, and academic freedom. AB - Elsevier Science refused to publish a study of IBM workers that IBM sought to keep from public view. Occupational and environmental health (OEH) suffers from the absence of a level playing field on which science can thrive. Industry pays for a substantial portion of OEH research. Studies done by private consulting firms or academic institutions may be published if the results suit the sponsoring companies, or they may be censored. OEH journals often reflect the dominance of industry influence on research in the papers they publish, sometimes withdrawing or modifying papers in line with industry and advertising agendas. Although such practices are widely recognized, no fundamental change is supported by government and industry or by professional organizations. PMID- 17915546 TI - Asbestos exposure causes mesothelioma, but not this asbestos exposure: an amicus brief to the Michigan Supreme Court. AB - Manufacturers of asbestos brakes, supported by many manufacturing and insurance industry amicus curie, requested the Michigan Supreme Court to dismiss testimony of an expert regarding the ability of asbestos dust from brakes to cause mesothelioma as "junk science". Scientists are concerned with the sweeping and unequivocal claims that any conclusion that asbestos from brakes caused a signature asbestos-related disease in a particular person must be "junk science". The manufacturers' sweeping pronouncements are what veer from accepted, reliable mainstream scientific methods and conclusions. This article outlines the evidence supporting the conclusion that asbestos from brakes can and does cause mesothelioma, and describes the defendants' attempts to fabricate doubt about this conclusion. PMID- 17915547 TI - Manipulating public health research: the nuclear and radiation health establishments. AB - Industry, government, and the military have systematically suppressed or manipulated epidemiologic research showing detrimental effects on human health from accidental or occupational exposures to ionizing radiation. This leads to conflicts of interest and compromised integrity among scientists in the radiation health establishment, it stifles dissemination of "unwelcome" findings and endangers public health. PMID- 17915548 TI - Malthusian pressures, genocide, and ecocide. AB - Historical models postulate that genocide cannot occur without the ideology and decisions of its authoritarian perpetrators and the indifference of bystanders. These models do not address genocidal risks from ecocide. Study objectives were to assess 1) the role of Malthusian pressures in recent genocides, 2) the role of ecocide and ecologic abuse in creating these pressures, and 3) strategies for prevention and deterrence. Analysis of reports, demographic studies, and time trends in recent genocides and recent ecocidal events from ecologic abuse suggests that Malthusian pressures and zero-sum rivalries over water, arable land, or natural resources by themselves do not lead to genocide. Such pressures may have exacerbated the political and socioeconomic predictors in Rwanda and Darfur, but not in former Yugoslavia. However, collapse of socioeconomic and governmental infrastructures following genocide can leave behind massive sustained damage to carrying capacity and sustainability. Surviving victims, if they return to their environments, will remain at risk for persecution. Ecocide- the large-scale destruction, depletion, or contamination of natural ecosystems- can result in widespread damage to health, survival, fertility, reproduction, and sustenance, and forced flight. International early warning and effective response systems are needed to deter or prevent political decisions to carry out genocide. Such systems must include long-term measures to resolve zero-sum conflicts over environmental resources and to prevent toxic risks to vulnerable populations and destruction of habitat by deliberate or wanton ecologic abuse, which itself should be redefined as a crime against humanity. PMID- 17915549 TI - New European community strategy for health and safety: the elephant in the room. AB - Although Lithuania has comprehensive health and safety legislation in line with EU requirements, on a range of general health and occupational health and safety (OHS) indicators, it is a poor performer. Survey data suggest that the norm for work in Lithuania is based on a regime of intensification without a participative working environment in which employees have a voice in safety management. Although European-style legislative reforms appear to be having no measurable effects on CHS performance in post-communist New Member States, the EU OHS strategy for 2007-2012 fails to take account of the deteriorated working environments in these states, suggesting that prospects for harmonization of working environment standards in the enlarged Europe may recede with eastward expansion. PMID- 17915550 TI - Industry influence and atrazine carcinogenicity. PMID- 17915551 TI - An inducible 70 kDa-class heat shock protein is constitutively expressed during early development and diapause in the annual killifish Austrofundulus limnaeus. AB - The annual killifish Austrofundulus limnaeus inhabits ephemeral ponds in regions of northern South America, where they survive the periodic drying of their habitat as diapausing embryos. These diapausing embryos are highly resistant to a number of environmental insults such as high temperature, dehydration, anoxia, and increased salinity. Molecular chaperones are known to play a role in stabilizing protein structure and function during events of cellular stress. Relative levels of heat shock protein (Hsp)70 were measured in developing and diapausing embryos of A. limnaeus using quantitative Western blots. An inducible or embryo-specific form of Hsp70 is expressed during embryonic development in A. limnaeus and is elevated during diapause II in this species. Constitutive expression of Hsp70 during development may afford these embryos protection from environmental stresses during development more quickly than relying on the induction of a classic heat shock response. PMID- 17915552 TI - Heat shock proteins in physiology and pathology: the Berlin meeting. PMID- 17915553 TI - Activation profiles of HSPA5 during the glomerular mesangial cell stress response to chemical injury. AB - Environmental injury has been associated with endoplasmic reticulum (ER) stress, a response characterized by activation of the unfolded protein response, proteasomal degradation of proteins, and induction of HSPA5, also known as GRP78 or BiP. Although HSPA5 has been implicated in the stress response to environmental injury in several cell types, its role in the glomerular ER stress response is unknown. In this study, we evaluated HSPA5 activation profiles in rat glomerular mesangial cells (rGMCs) challenged with heavy metals (HgCl2 or Pb2+ acetate) or polycyclic aromatic hydrocarbons (PAHs, ie, benzo(a)pyrene [BaP]). Challenge of rGMCs with 1 or 10 microM HgCl2 or Pb2+ acetate increased HSPA5 mRNA and protein levels. The induction response was sensitive to transcriptional and translational inhibition by actinomycin D (AD) and cyclohexamide, respectively. HSPA5 mRNA was induced by 3 microM BaP in an AD-sensitive manner, but this response was unaffected by the presence of heavy metals. A promoter construct containing sequences that mediate thapsigargin (TH) inducibility of the HSPA5 promoter was refractory to both heavy metals and BaP. The HSPA5 induction response in rGMCs is conserved because it was reproduced with fidelity in immunolocalization experiments of HSPA5 protein in M15 and HEK293 cells in embryonic lines of murine and human origin, respectively. Collectively, these findings identify HSPA5 in the stress response of rGMCs and implicate regulatory mechanisms that are distinct from those involved in TH inducibility. PMID- 17915554 TI - Hsp70B' regulation and function. AB - Heat shock protein (Hsp) 70B' is a human Hsp70 chaperone that is strictly inducible, having little or no basal expression levels in most cells. Using siRNAs to knockdown Hsp70B' and Hsp72 in HT-29, SW-480, and CRL-1807 human colon cell lines, we have found that the two are regulated coordinately in response to stress. We also have found that proteasome inhibition is a potent activator of Hsp70B'. Flow cytometry was used to assay Hsp70B' promoter activity in HT-29eGFP cells in this study. Knockdown of both Hsp70B'- and Hsp72-sensitized cells to heat stress and increasing concentrations of proteasome inhibitor. These data support the conclusion that Hsp72 is the primary Hsp70 family responder to increasing levels of proteotoxic stress, and Hsp70B' is a secondary responder. Interestingly ZnSO4 induces Hsp70B' and not Hsp72 in CRL-1807 cells, suggesting a stressor-specific primary role for Hsp70B'. Both Hsp70B' and Hsp72 are important for maintaining viability under conditions that increase the accumulation of damaged proteins in HT-29 cells. These findings are likely to be important in pathological conditions in which Hsp70B' contributes to cell survival. PMID- 17915555 TI - Association of increased heat shock protein 70 levels in the lymphocyte with high risk of adverse pregnancy outcomes in early pregnancy: a nested case-control study. AB - Studies suggest that heat shock proteins (Hsps), Hsp70 in particular, may play a role in embryogenesis and reproduction. As the first trimester is the critical period of human fetal development, we tested whether there is an association between Hsp70 expression in lymphocytes and adverse pregnancy outcomes (APOs) during that period. We measured lymphocyte Hsp70 levels by immunoblot in 55 pregnant women with APOs and 110 well-matched controls selected from 778 pregnant women in a nested case-control study. Multivariate logistic regression models were used to investigate the association between lymphocyte Hsp70 levels and risk of developing APOs. Our data showed that Hsp70 levels in women with APOs, especially those younger than 29 years old, were significantly higher than controls (193 vs 135 units, P < 0.001) and that the elevated Hsp70 levels were associated with a significantly increased risk of APOs (adjusted OR = 1.014; 95% CI = 1.008-1.020, P < 0.001). Our results also showed that the sensitivity, specificity, and positive and negative predictive values were 78%, 60%, 50%, and 85%, respectively, among these pregnant women. Adjusted ORs and 95% CI for the association between a Hsp70 value > 153 IOD and APOs were statistically significant (OR = 8.78, 95% CI = 2.79-27.64, P < 0.001). These results suggest that Hsp70 may play a role in the etiology of APOs. However, the underlying mechanisms for the elevation of Hsp70 in women with APOs and whether Hsp70 can be applied as a clinical indicator of APOs warrant further investigations. PMID- 17915556 TI - Induction of heat shock proteins in differentiated human and rodent neurons by celastrol. AB - Neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis have been termed protein misfolding disorders that are characterized by the neuronal accumulation of protein aggregates. Manipulation of the cellular stress-response involving induction of heat shock proteins (Hsps) in differentiated neurons offers a therapeutic strategy to counter conformational changes in neuronal proteins that trigger pathogenic cascades resulting in neurodegenerative diseases. Hsps are protein repair agents that provide a line of defense against misfolded, aggregation-prone proteins. These proteins are not induced in differentiated neurons by conventional heat shock. We have found that celastrol, a quinine methide triterpene, induced expression of a wider set of Hsps, including Hsp70B', in differentiated human neurons grown in tissue culture compared to cultured rodent neuronal cells. Hence the beneficial effect of celastrol against human neurodegenerative diseases may exceed its potential in rodent models of these diseases. PMID- 17915557 TI - Heat shock protein 70 regulates cellular redox status by modulating glutathione related enzyme activities. AB - Heat shock protein (Hsp) 70 has been reported to protect various cells and tissues from ischemic damage. However, the molecular mechanisms of the protection are incompletely understood. Ischemia induces significant alterations in cellular redox status that plays a critical role in cell survival/death pathways. We investigated the effects of Hsp70 overexpression on cellular redox status in Madin-Darby canine kidney (MDCK) cells under both hypoxic and ischemic conditions with 3 different approaches: reactive oxygen species (ROS) measurement by a fluorescence probe, redox environment evaluation by a hydroxylamine spin probe, and redox status assessment by the glutathione/glutathione disulfide (GSH/GSSG) ratio. Results from each of these approaches showed that the redox status in Hsp70 cells was more reducing than that in control cells under either hypoxic or oxygen and glucose deprivation (OGD) conditions. In order to determine the mechanisms that mediated the alterations in redox state in Hsp70 cells, we measured the activities of glutathione peroxidase (GPx) and glutathione reductase (GR), two GSH-related antioxidant enzymes. We found that OGD exposure increased GPx and GR activities 47% and 55% from their basal levels (no stress) in Hsp70 cells, compared to only 18% and 0% increase in control cells, respectively. These data, for the first time, indicate that Hsp70 modulates the activities of GPx and GR that regulate cellular redox status in response to ischemic stress, which may be important in Hsp70's cytoprotective effects. PMID- 17915558 TI - Developmentally regulated synthesis of p8, a stress-associated transcription cofactor, in diapause-destined embryos of Artemia franciscana. AB - Diapause-destined embryos of the crustacean Artemia franciscana arrest as gastrulae, acquire extreme stress tolerance, and enter profound metabolic dormancy. Among genes upregulated at 2 days postfertilization in these embryos is a homologue of p8, a stress-inducible transcription cofactor. Artemia p8 is smaller than vertebrate homologues but shares a basic helix-loop-helix domain and a bipartite nuclear localization signal. Probing of restriction digested DNA on Southern blots indicated a single Artemia p8 gene and 5'-RACE specified 2 transcription start sites. Several putative cis-acting regulatory sequences, including two heat shock elements, appeared upstream of the p8 transcription start site. Artemia p8 mRNA increased sharply at 1 day postfertilization in diapause-destined embryos and then declined, whereas p8 protein appeared 2 days postfertilization and remained relatively constant throughout development, indicating a stable protein. p8 was not detectable in nauplius-destined (nondiapause) Artemia embryos. Immunofluorescent staining revealed p8 within Artemia nuclei. The results support the idea that p8, a known stressresponsive transcription cofactor, mediates gene expression in diapause-destined Artemia embryos. p8 is the first diapause-related transcription factor identified in crustaceans and 1 of only a small number of such proteins identified in any organism undergoing diapause. PMID- 17915559 TI - Circulating heat shock proteins and inflammatory markers in patients with idiopathic left ventricular dysfunction: their relationships with myocardial and microvascular impairment. AB - Little information is available on peripheral levels of Hsp72, Hsp60, and anti Hsp60 antibodies in patients with left ventricular (LV) dysfunction due to non atherosclerotic cardiac disease. In this study, serum Hsp72, Hsp60 and anti-Hsp60 antibodies, IL-6, and C-reactive protein (CRP) were measured in 44 healthy controls and in 82 patients with angiographically normal coronary arteries (LV ejection fraction [EF] > or = 50%, n=22; -35% to <50%, n=32; <35%, n=28). Patients with more severe disease (more depressed myocardial blood flow at rest and during dipyridamole, indicative of coronary microvascular impairment) showed more elevated circulating Hsp60 and auto-antibodies, Hsp72, and CRP levels. IL-6 was increased progressively as a function of severity of LV dysfunction. Anti Hsp60 antibodies, Hsp72, and IL-6 were significantly correlated with brain natriuretic peptide (BNP) levels and LV end-diastolic dimensions (LVEDD) values. IL-6 tended to be related with Hsp72 in particular in patients with more severe disease (r = 0.45, P = 0.021). Hsp60 and Hsp72 activation and inflammatory markers were correlated with the extent of cardiac and microvascular dysfunction in patients with angiographycally normal coronary arteries. These results suggest a pathogenic role of infective-metabolic insult and inflammatory reaction in the development of vascular and myocardial damage in patients with heart failure even in the absence of overt coronary artery disease. PMID- 17915560 TI - Expression of heat shock protein 70 in the thermally stressed antarctic clam Laternula elliptica. AB - Heat shock protein 70 (designated Laternula elliptica Hsp70 (LEHsp70)) expression was investigated in an Antarctic mud clam to see whether or not the inducible heat shock response has been conserved throughout over 25 million years of adaptation to constant low environmental temperatures. LEHsp70 cDNA was cloned and sequenced from the Antarctic clam Laternula elliptica. We used degenerated primers designed in the highly conserved regions of Hsp to amplify the corresponding mRNA, and full-length cDNA was obtained by rapid amplification of cDNA ends (RACE). The full length of LEHsp70 cDNA was 2470 bp, with a 5' untranslated region (UTR) of 92 bp, a 3' UTR of 416 bp, and an open reading frame (ORF) of 1962 bp encoding a polypeptide of 653 amino acids with an estimated molecular mass of 71.266 kDa and an estimated isoelectric point of 5.20. LEHsp70 contained highly conserved functional motifs of the cytosolic Hsp70 family. Expression of the LEHsp70 gene was quantified by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) of digestive gland and gill tissues. Heat shock (10 degrees C for different time periods) caused rapid induction of LEHsp70. A significant 4.6 +/- 0.14-fold increase in the LEHsp70/beta-Actin mRNA ratio occurred in the gill at 12 hours, which returned to baseline after 48 hours. In contrast, the maximum expression in the digestive gland (3.6 +/- 0.36) was reached at 24 hours and was still significant after 48 hours (1.89 +/- 0.21). This indicates that LEHsp70 may play an important role in mediating thermal stress and tolerance in this clam. PMID- 17915562 TI - Clinical semiology of neuromuscular diseases. Bent spine myopathy or syndrome. PMID- 17915561 TI - HSF2 binds to the Hsp90, Hsp27, and c-Fos promoters constitutively and modulates their expression. AB - Although the vast majority of genomic DNA is tightly compacted during mitosis, the promoter regions of a number of genes remain in a less compacted state throughout this stage of the cell cycle. The decreased compaction of these promoter regions, which is referred to as gene bookmarking, is thought to be important for the ability of cells to express these genes during the following interphase. Previously, we reported a role for the DNA-binding protein heat shock factor (HSF2) in bookmarking the stress-inducible 70,000-Da heat shock protein (hsp70) gene. In this report, we have extended those studies and found that during mitosis, HSF2 is bound to the HSE promoter elements of other heat shock genes, including hsp90 and hsp27, as well as the proto-oncogene c-fos. The presence of HSF2 is important for expression of these genes because blocking HSF2 levels by RNA interference techniques leads to decreased levels of these proteins. These results suggest that HSF2 is important for constitutive as well as stress-inducible expression of HSE-containing genes. PMID- 17915563 TI - The emerging diversity of neuromuscular junction disorders. AB - Research advances over the last 30 years have shown that key transmembrane proteins at the neuromuscular junction are vulnerable to antibody-mediated autoimmune attack These targets are acetylcholine receptors (AChRs) and muscle specific kinase (MuSK) in myasthenia gravis, voltage-gated calcium channels (VGCCs) in the Lambert-Eaton myasthenic syndrome (LEMS), and voltage-gated potassium channels (VGKCs) in neuromyotonia. In parallel with these immunological advances, mutations identified in genes encoding pre-synaptic, synaptic and postsynaptic proteins that are crucial to neuromuscular transmission have revealed a similar diversity of congenital myasthenic syndromes (CMS). These discoveries have had a major impact on diagnosis and management. PMID- 17915564 TI - Axial myopathies: an elderly disorder. PMID- 17915566 TI - Muscular dystrophy, incurability, eugenics. AB - The medical entity "muscular dystrophy" has been the object of a recent opinion campaign aimed at promoting a law in favour of euthanasia. This disease has become, in the eyes of the public, a media model of a particularly severe and incurable disease. This very widespread statement does not correspond to reality as far as concerns the life of these patients, to the condition that they have benefited from a very useful and fully provided empirical treatment. As already seen, the hope for life has already doubled, without clear limits. The idea of inducing an interruption when at death's door, as long as a systematic prevention prior to birth, does not conform with the motivated opinion of the majority of patients consulted. On the contrary, the dogma of incurability may lead to dramatic individual consequences which should be stressed, from a medical viewpoint, on account of the unacceptable risks of social injustice or eugenics that this would imply. PMID- 17915565 TI - Markers of oxidative stress and aging in Duchene muscular dystrophy patients and the possible ameliorating effect of He:Ne laser. AB - Replicative aging and oxidative stress are two plausible theories explaining the etiology of muscular dystrophy. The first theory indicates that replicative aging of myogenic cells (satellite cells), owing to enhanced myofiber turnover, is a plausible explanation of the progression of Duchenne muscular dystrophy (DMD). The oxidative stress theory indicates that failure of muscle regeneration to keep up with the ongoing apoptosis and necrosis following oxidative stress, that normally associates muscular exercise, leads to muscle atrophy in DMD. To test for these two theories, markers of replicative aging and oxidative stress were assessed in the blood of 30 DMD patients vs. 20 normal healthy age matching controls. Markers of replicative aging showed significantly lower telomerase activity, significantly increased expression of receptors for advanced glycation end products (RAGEs) mRNA and Bax mRNA (an apoptotic gene) in DMD compared to controls. There was a significant increase in markers of oxidative stress among DMD patients compared to controls, measured in terms of increased apoptotic percentage in circulating mononuclear cells, increased lipid peroxidation measured in terms of plasma malondialdehyde (MDA) and increased protein carbonyls. Levels of plasma nitric oxide (NO), which neutralizes oxygen radicals, and expression of inducible nitric oxide synthase (iNOS) mRNA in neutrophils was significantly lower among DMD compared to controls. Biostimulation of WBC by helium neon (He:Ne) laser irradiation induced a significant increase in the expression of iNOS mRNA and plasma NO levels, but still at a lower level compared to controls. He:Ne laser irradiation induced a marked decrease in markers of oxidative stress among DMD patients compared to their level before irradiation, measured in terms of decreased plasma protein carbonyls, decreased plasma MDA, and decreased apoptosis percentage. CONCLUSION: This study points to that oxidative stress is the prime cause for muscle degeneration in DMD and points out to the possible ameliorative effect of He:Ne laser on this PMID- 17915567 TI - Muscle glycogenoses: an overview. PMID- 17915568 TI - Glycogen storage disease type II: clinical overview. AB - Glycogen storage disease type II has a broad continuous clinical spectrum in terms of onset, involvement of organs and life expectancy. Infantile onset is the most severe form, presenting with prominent cardiomyopathy, hypotonia, hepatomegaly and death before 12 months of life. Late onset form has onset at any age, lack of severe (or absence of) cardiac involvement, progressive skeletal muscle dysfunction and less dismal short-term prognosis. In addition to muscle and heart involvement, other tissues are affected liver, spleen, endothelium, lung, brain, anterior horns, peripheral nerves. In fact some patients with infantile form have hearing loss, abnormal brain myelination and central fever and some adult patients show aneurysms of brain arteries due to accumulation of glycogen in vessels. As for other treatable lysosomal diseases, the advent of enzyme replacement therapy will change the natural history of this disease and also will increase our knowledge concerning clinical heterogeneity. PMID- 17915569 TI - Role of autophagy in the pathogenesis of Pompe disease. AB - In Pompe disease, a deficiency of lysosomal acid alpha-glucosidase, glycogen accumulates in multiple tissues, but clinical manifestations are mainly due to skeletal and cardiac muscle involvement. A major advance has been the development of enzyme replacement therapy (ERT), which recently became available for Pompe patients. Based on clinical and pre-clinical studies, the effective clearance of skeletal muscle glycogen appears to be more difficult than anticipated. Skeletal muscle destruction and resistance to therapy remain unsolved problems. We have found that the cellular pathology in Pompe disease spreads to affect both the endocytic and autophagic pathways, leading to excessive autophagic buildup in therapy resistant muscle fibers of knockout mice. Furthermore, the autophagic buildup had a profound effect on the trafficking and processing of the therapeutic enzyme along the endocytic pathway. These findings may explain why ERT often falls short of reversing the disease process, and point to new avenues for the development of pharmacological intervention. PMID- 17915570 TI - Progress and problems when considering gene therapy for GSD-II. PMID- 17915571 TI - McArdle disease: molecular genetic update. AB - McArdle disease or Glycogenosis type V is an autosomal recessive metabolic disorder caused by a deficiency of the muscle isoform of glycogen phosphorylase (myophosphorylase, PYGM), the specific skeletal muscle enzyme that initiates glycogen breakdown. Since the first clinical description by Brian McArdle in 1951, several patients have been identified worldwide and significant advances have been made in the study of molecular genetics of the disease. Molecular heterogeneity has been demonstrated by the identification to date of more than 65 mutations in the PYGM gene. PMID- 17915572 TI - Pharmacological and nutritional treatment trials in McArdle disease. AB - A systematic review of evidence for randomised controlled trials using pharmacologic and nutritional therapies in McArdle disease was undertaken. Primary outcome measures included any objective assessment of exercise endurance. Secondary outcome measures included changes in metabolic parameters, subjective measures such as quality of life scores and adverse outcomes. Ten randomised controlled trials were identified. Two trials low dose creatine (60 mg/kg/day) and oral sucrose 75 g prior to exercise demonstrated a positive effect. PMID- 17915574 TI - Chronic therapy for McArdle disease: the randomized trial with ACE inhibitor. PMID- 17915573 TI - Treatment of glycogenosys type V (McArdle disease) with creatine and ketogenic diet with clinical scores and with 31P-MRS on working leg muscle. AB - McArdle's disease is caused by genetic defects of the muscle-specific isozyme of glycogen phosphorylase, which block ATP formation from glycogen in skeletal muscle. Creatine supplementation and ketogenic diet have been tested as potential supplements for muscle energy metabolism which may improve muscle symptomatic. Outcome measures were clinical scores describing muscle symptomatic and parameters derived from 31P-MRS examinations on working muscle. In two placebo controlled cross-over studies low dose creatine showed beneficial effects on muscle symptoms and performance whereas high dose creatine distinctly worsened muscle symptomatic in patients. In both studies, however, the absence of an elevation in phosphocreatine indicated the absence of a creatine uptake by the muscle fibre. The effects of creatine on muscle symptomatic may be independent from energy metabolism in muscle. In a case study, ketogenic diet improved muscle symptomatic and performance. However, these effects again did not result in 31P MRS visible changes in muscle energy metabolism. PMID- 17915575 TI - Molecular genetics of late onset glycogen storage disease II in Italy. AB - Glycogen Storage Disease Type II (GSDII) is a recessively inherited disorder due to the deficiency of acid alpha-glucosidase (GAA) that results in glycogen accumulation in the lysosomes. The molecular analysis of the GAA gene was performed on 45 Italian patients with late onset GSDII. DHPLC analysis revealed 28 polymorphisms spread all over the GAA gene. Direct sequencing identified the 96% of the mutant alleles, 12 of which are novel. Missense mutations were functionally characterized by enzyme activity and protein processing in a human GAA deficient cell line while splicing mutations were studied by RT-PCR and in silico analysis. A complex allele was also identified carrying three different alterations in cis. All the patients studied carried a severe mutation in combination with a milder one, which explains the late onset of the disease. The c.-32-13T > G was the most frequent mutation, present as compound heterozygote in 85% of the patients as described in other late onset GSDII Caucasian populations. Interestingly, 10 of the 45 patients carried the c.-32-13T > G associated to the severe c.2237G > A (p.W746X) mutation. However, despite the common genotype, patients presented with a wide variability in residual enzyme activity, age of appearance of clinical signs and rate of disease progression, suggesting that other genetic/environment factors may modulate clinical presentation. PMID- 17915576 TI - Clinical, biochemical and genetic features of glycogen debranching enzyme deficiency. AB - Deficiency of debrancher enzyme causes Glycogen Storage Disease (GSD) type III, an autosomal recessive disorder, characterized by tissue accumulation of abnormally structured glycogen. This report reviews current clinical and molecular knowledge about this disorder and describes the variability at phenotype and genotype levels of a large group of Italian GSDIII patients. PMID- 17915577 TI - Neuromuscular forms of glycogen branching enzyme deficiency. AB - Deficiency of glycogen branching enzyme is causative of Glycogen Storage Disease type IV (GSD-IV), a rare autosomal recessive disorder of the glycogen synthesis, characterized by the accumulation of amylopectin-like polysaccharide, also known as polyglucosan, in almost all tissues. Its clinical presentation is variable and involves the liver or the neuromuscular system and different mutations in the GBE1 gene, located on chromosome 3, have been identified in both phenotypes. This review will addresses the neuromuscular clinical variants, focusing on the molecular genetics aspects of this disorder. PMID- 17915578 TI - LAMP-2 deficiency (Danon disease). PMID- 17915579 TI - Lafora disease, seizures and sugars. AB - Lafora disease (LD) is the most severe form of Progressive Myoclonus Epilepsy with teenage onset. It has an autosomal recessive mode of inheritance and is almost universally fatal by the second or third decade of life. To date, there is no prevention or cure. In the last decade, with the identification of the genes responsible for this disease, much knowledge has been gained with the potential for the future development of effective treatment. This review will briefly address clinical issues and will focus on the molecular aspects of the disease. PMID- 17915580 TI - Update on treatment of lysosomal storage diseases. AB - Lysosomal storage diseases (LSDs) are a large group of disorders caused by a deficiency of specific enzymes responsible for the degradation of substances present in lysosomes. In the past few years, treatments for LSDs were non specific and could only cope with signs and symptoms of the diseases. A successful therapeutic approach to LSDs should instead address to the underlying causes of the diseases, thus helping the degradation of the accumulated metabolites in the various organs, and at the same time preventing their further deposition. One way is to see to an available source of the deficient enzyme: bone marrow transplantation, enzyme replacement therapy and gene therapy are based on this rationale. The purpose of substrate reduction therapy is to down regulate the formation of the lysosomal substance to a rate at which the residual enzyme activity can catabolize the stored and de novo produced lysosomal substrate. Chemical chaperone therapy is based on small molecules able to bind and stabilize the misfolded enzymes. This paper offers a historical overview on the therapeutic strategies for LSDs. PMID- 17915581 TI - [The treatment of the exsudative age-related macular degeneration with choroidal neovascular membrane, its possibilities and economical indexes]. AB - AIM: To establish the probable prevalence and incidence of the exsudative (wet) age-related macular degeneration (AMD) in the Czech Republic and to compare possibilities and the expensiveness of the photodynamic therapy (PDT) and the treatment with substances blocking the vascular endothelial growth factor (anti VEGF). METHODS: The calculation of the probable prevalence and incidence of the exsudative AMD in the Czech Republic (CR) was based on the world epidemiological studies. The expenses of the PDT and the anti-VEGF treatment for the patient and for the health insurances were based on the calculation of the Visudyne and anti VEGF substances' prices in CR and worldwide. RESULTS: If the European prevalence of the exsudative AMD in patients over 65 years of age is 2.3% and the prevalence of the risk soft drusen is 15 %, so in the CR out of 1.44 millions of inhabitants older than 65 years have approximately 33 000 the AMD, and the drusen at risk 216.000 inhabitants. If the 5-years incidence of the exsudative AMD in patients with drusen at risk is 3.4%, it may be expected the turn of the soft drusen into the exsudative AMD during the 5-years period in 7340 patients, or 1460 patients a year. The PDT is indicated in approx. 20% of exsudative AMD with the classical or predominantly classical choroid neovascularization (CNV), i.e. approx. 300 patients a year. If, at the average, 5.6 PDT sessions during 2 years are needed and the price is 42.500 CZK (Czech Crowns; 1 USD = approx. 20-22 CZK; 1 Euro = approx. 27-29 CZK) for one injection of Visudyne, so the average cost per patient is 238,000 CZK and the costs for all of them 71.4 millions of CZK. If the patient's participation is 7395 CZK per one dose of Visudyne, then the average patient's expense for the PDT during the 2 years period is 41 412 CZK. Anti-VEFG drugs as intravitreal injections are effective in all forms of exsudative AMD. Macugen (pegaptanib), already registered in the CR, should be applied in six weeks intervals, during the two-years treatment period altogether up to 17 injections. At the price of 1000 USD for one application, the average treatment cost is 17,000 USD (= approx. 350, 000 CZK). If the one-year incidence of exsudative AMD is 1460 patients,their treatment with Macugen would cost 511 mil. CZK, and the treatment of 1140 patients with occult CNV not suitable for PDT would cost 410 mil. CZK. Lucentis (ranibizumab), which is about to be registered in the CR, is applied in one-months intervals, during two years altogether up to 20 injections. At the price 1200 USD a dose, the treatment costs for one patient would be 24,000 USD (approx. 500 000 CZK), treatment of all patients with exsudative AMD 730 mil. CZK, and costs for patients not suitable-for PDT treatment would be 585 mil. CZK. The intravitreal application of Avastin (bevacizumab) is "off label"; in the first three months is applied monthly, and later on, in 1-3 months interval, until the disappearance of exsudative changes. The maximum of applications is 10 injections during 2 years. At the price 3000 CZK for 1 injection, the treatment costs for one patient would be 30,000 CZK; treatment costs for all patients would be 43.8 mil. CZK, and for patients not suitable for PDT would be 34.8 mil. CZK. CONCLUSION: Nowadays, the PDT and anti VEFG substances are the optimal treatment methods in exsudative AMD with CNV. PDT i s indicated in theclassical or predominantly classical choroid neovascularization (CNV), anti-VEFG drugs are effective in all forms of exsudative AMD. The high prices of Visudyne, Macugen, and Lucentis create barriers to their general use in all patients with exsudative AMD. Ten times lower price of "off label" applied Avastin supports its use in the competition with other anti-VEGF drugs. The combination of PDT with anti-VEGF drugs suggests the possible way, how to, not only, improve the effectiveness, but also to lower the number of applications, and by this means, also to reduce the economical expenses of the treatment dramatically. PMID- 17915582 TI - [Intraocular pressure (IOP) in rabbits after application of amino acids taurine and arginine with betablocker timolol]. AB - PURPOSE: To assess the efficiency of the antiglaucomatic 0.5 % Timolol mixture with two amino acids: (1) 10% L-taurine and (2) 10% L-arginine on the physiological IOP in rabbits. METHODS: Into the left conjunctival sac of 5 adult female rabbits of the New Zealand White species was instilled (a) 0.5% Timolol alone; (b) the mixture of 10 % L-arginine and 10% L-taurine in aqua pro injectione; (c) the mixture of 10% L-arginine and 10% L-taurine in 0.5% Timolol. The IOP was measured before and 5, 15, 30, 60, 120, 180, 240 min., and 24 hours after the instillation. The right eye was used as control. RESULTS: (a) The mixture of the amino acids arginine and taurine in Timolol achieved significant IOP decrease in rabbits (except results at 5 min. and 24 hours after instillation) in all measured times with the maximum in 120 and 180 min. showing the biphasic character of the IOP decrease; (b) the mixture of amino acid taurine and arginine reached the border of significance (p < 0.05). The IOP decrease showed biphasic character, with the maximum in 60 min. (decrease of 3.8 mm Hg) and in 120 min. (decrease of 3.3 mm Hg); (c) the antiglaucomatic drops 0.5% Timolol alone decreased the IOP values without significance comparing with the control eye. The maximum decrease was measured in 120 min. (2.9 mm Hg). CONCLUSIONS: The mixture of amino acids taurine and arginine in 0.5% Timolol proved significant decrease of the IOP physiological values in rabbits. The maximum decrease was achieved 120, 180 min., and 24 hours after the instillation. The average value of the decrease 24 hours after instillation, compared with the control eye, was 3.2 mm Hg. PMID- 17915583 TI - [The importance of HRT II (Heidelberg retina tomography) and perimeter Octopus 101 in glaucoma diseases]. AB - PURPOSE: To follow up and statistically evaluate the importance of HRT II (Heidelberg retina tomograph) and perimeter Octopus 101 in glaucoma diseases. MATERIAL AND METHODS: There were 154 eyes (77 patients) enrolled into prospective study. The follow up period was 3 years, from January 2003 to December 2005. Patients were divided into 3 groups: (1) the primary open angle glaucoma (PGOU) group and the preperimetric form of PGOU group (88 eyes), (2) the ocular hypertension (OH) group with medical therapy and without medical therapy (44 eyes) and (3) the group of persons with large optic disc areas (22 eyes). The structural optic nerve head (ONH) changes were interpreted by means of HRT II equipment and the functional changes were evaluated by means of perimeter Octopus 101 every 6 months. The statistic analysis was performed with the program Statgraphics plus. RESULTS: HRT II is a reproducible method to follow up the structural changes of the ONH for its low variability of achieved results in all evaluated groups. Low correlation between MD (mean defect) and CLV (corrected lose variance) parameters in the first and the third groups occurred. The standard automated perimetry was accompanied by greater long-term variability, making it less reproducible, but it is useful method comparing to HRT II. In the group of PGOU patients, significant correlations between MD perimetric parameter and the mean RNFL (retinal nerve fibre layer) thickness parameters were observed by means of HRT II. CONCLUSION: HRT II is not able to replace visual field examination with perimetry, both methods are important in the evaluation of glaucoma patients. PMID- 17915584 TI - [Importance of structural examination methods in the follow-up of patients with ocular hypertension]. AB - PURPOSE: To evaluate the contribution of Heidelberg Retina Tomograph (HRT) and blue-on-yellow perimetry (B-Y) for the follow-up and early diagnosis of glaucoma in patients with ocular hypertension (OHT). We also dealed with the comparison of visual field (VF) examination by means of the B-Y and white-on-white perimetry, correlation between VF indexes and HRT parameters, importance of the central corneal thickness (CCT) and the comparison of other characteristics of the study and control groups during the 3-year-study-period. PATIENTS AND METHODS: Both eyes of 28 patients with OHT (study group, SG) and 15 control subjects (control group, CG) were included in the study. Subjects in both groups were examined annually. At each visit complete ophthalmological examination was carried out, including the biomicroscopic evaluation of the optic disc, aplanation tonometry, HRT II analysis of the optic nerve head (ONH) and B-Y perimetry. Pachymetry was performed only once at the beginning of the study. RESULTS: During the 3-year follow-up structural changes of the ONH developed in 2 patients with OHT only. They were not accompanied by VF defects on B-Y perimetry. Intraocular pressure and CCT were higher in the SG (p < 0.01). CCT significantly and positively correlated with the level of intraocular pressure in both groups. Mean sensitivity (MS) of the B-Y VF was repeatedly smaller in the SG compared to the CG (p < 0.01). Difference between MS a mean defect (MD) of the B-Y and white-on white VF in the SG was significant (p < 0.01). The B-Y perimetry test lasted much longer than standard perimetry. A marked learning effect was found with B-Y perimetry. Despite significant correlations of follow-up B-Y perimetry examinations, high test-retest variability was found. No significant change in topographic parameters of the ONH occurred during the follow-up period. HRT displayed low variability of follow-up examinations. No significant correlation between topographic parameters of the ONH and VF indexes on B-Y perimetry was found. CONCLUSION: In 2 cases of OHT the HRT preceded VF changes on B-Y perimetry. HRT showed high reproducibility, on the contrary, B-Y perimetry was accompanied by high variability making the distinction of fluctuation and progression more difficult. Pachymetry is a very useful tool in patients with OHT. PMID- 17915585 TI - [Transmissing electron microscopy of the vitreo-macular border in clinically significant diabetic macular edema]. AB - The authors examined samples of the epimacular tissue in clinically significant macular edema by means of the transmissing electron microscopy. They did not found morphological differences between samples from patients already treated by means of laser photocoagulation before the pars plana vitrectomy and those without the laser treatment. Findings may be divided into three groups: (1) the inner limiting membrane (ILM) covered with collagen vitreous fibers, (2) cells' elements of the fibroblasts category, and (3) fibrous astrocytes in the vitreous cortex constituting one- or multilayer cellular membranes. PMID- 17915586 TI - [First experiences with the Visante apparatus]. AB - The apparatus Visante (Zeiss) was tested on the group of 37 eyes with selected findings. This apparatus enables to visualize the anterior segment of the eye by means of OCT imaging. The main asset of the new imaging method is the accuracy of the image and the possibility to depict exactly the corneo-scleral angle and the artificial structures in the anterior chamber of the eye. The main advantage of the new apparatus is especially the imaging and measurement of the anterior segment structures of the eye in glaucoma and artificial structures in the anterior chamber and corneo-scleral angle of the eye PMID- 17915587 TI - [Transcorneal and transscleral iontophoresis of the dexamethasone phosphate into the rabbit eye]. AB - PURPOSE: To evaluate the efficiency of the dexamethasone phosphate penetration into the rabbit eye after transcorneal and transscleral iontophoresis using a drug loaded hydrogel assembled on a portable iontophoretic Mini Ion device. METHODS: lontophoresis of dexamethasone phosphate was studied in healthy rabbits using drug-loaded disposable HEMA hydrogel sponges and portable iontophoretic device. Corneal iontophoretic administration was performed with electric current of 1 mAmp for 1, 2, and 4 min. In the control group, the dexamethasone was applied in drops into the conjunctival sac. Transconjunctival and transscleral iontophoresis were performed in the pars plana area, through the conjunctiva or directly on the sclera. Dexamethasone concentrations were assayed using HPLC method. To study the anatomical changes after iontophoresis application, histological examinations of corneas excised 5 minutes and 8 hours after the procedure were performed. RESULTS: Dexamethasone levels in the rabbits' corneas after a single transcorneal iontophoresis were up to 38 times higher compared to those obtained after topical eye drops instillation. High drug concentrations were obtained in the retina and sclera 4 hours after transscleral iontophoresis as well. There were no statistically significant differences in the drug concentration after transscleral and tranconjunctival iontophoresis. Histological examination of the corneas after the iontophoresis showed only discrete reversible changes of the epithelium and the stroma. CONCLUSION: A short, low current, non-invasive iontophoretic treatment using the dexamethasone-loaded hydrogels has a potential clinical value in increasing the drug's penetration into the anterior and posterior segment of the eye. PMID- 17915588 TI - [Diagnostic and therapeutic vitrectomy in uveitis]. PMID- 17915589 TI - The high-performance dentist. PMID- 17915590 TI - Strategies for successful esthetic dental treatment. AB - The foundational principles of esthetic smiles reveal the direct influence of individual tooth alignment on dentofacial relationships. The use of clinical photography is an essential means to identify esthetic problems. Smile design provides an opportunity for effective communication to discuss treatment alternatives with the patient in the consultation process. The scope of treatment can be determined, and treatment limitations can be explained. Smile design findings influence preparation design, material selection, and laboratory communication for enhanced predictability and improved treatment success. PMID- 17915591 TI - Clinical crown lengthening in the esthetic zone. AB - Periodontal surgical procedures consisting of gingival flaps and osseous recontouring are indicated for crown lengthening of several contiguous teeth in the esthetic zone; both in cases where restorations are required and in cases where no restorations are planned, such as in patients with excessive gingival display due to altered passive eruption. Forced tooth eruption via orthodontic extrusion is the technique of choice when clinical crown lengthening is necessary on isolated teeth in the esthetic zone. PMID- 17915592 TI - Etiology and management of whitening-induced tooth hypersensitivity. AB - Tooth hypersensitivity has long been, and continues to be, the most commonly reported adverse effect of vital tooth whitening with peroxide gels. The complex etiology of whitening-induced tooth hypersensitivity has been a major obstacle in developing a definitive strategy for its prevention. This article reviews the multiple etiologic factors implicated in whitening-induced tooth hypersensitivity and the evidence for efficacy of various strategies for its management. PMID- 17915593 TI - Perspectives on the 2007 AHA Endocarditis Prevention Guidelines. AB - From 2005 to 2007, the American Heart Association convened a consensus panel of experts to revisit the guidelines for the premedication of patients with cardiac defects prior to dental treatment. Presented in this article is a summary of the guidelines as well as commentary on the process. PMID- 17915594 TI - No apologies! PMID- 17915595 TI - Pilot performance, strategy, and workload while executing approaches at steep angles and with lower landing minima. AB - OBJECTIVE: We examined the willingness and ability of general aviation pilots to execute steep approaches in low-visibility conditions into nontowered airports. BACKGROUND: Executing steep approaches in poor weather is required for a proposed Small Aircraft Transportation System (SATS) that consists of small aircraft flying direct routes to a network of regional airports. METHOD: Across two experiments, 17 pilots rated for Instrument Flight Rules at George Mason University or Virginia Tech flew a Cessna 172R simulator into Blacksburg, Virginia. Pilots were familiarized with the simulator and asked to fly approaches with either a 200- or 400-foot ceiling (at approach angles of 3 degrees, 5 degrees, and 7 degrees in the first experiment, 3 degrees and 6 degrees in the second). Pilots rated subjective workload and the simulator recorded flight parameters for each set of approaches. RESULTS: Approaches with a 5 degree approach angle produced safe landings with minimal deviations from normal descent control configurations and were rated as having a moderate level of workload. Approaches with 6 degree and 7 degree approach angles produced safe landings but high workload ratings. Pilots reduced power to control the speed of descent and flew the aircraft slightly above the glide path to gain time to control the landing. CONCLUSION: Although the 6 degree and 7 degree approaches may not be practical for routine approaches, they may be achievable in the event of an emergency. APPLICATION: Further work using other aircraft flying under a wider variety of conditions is needed before implementing SATS-type flights into airports intended to supplant or complement commercial operations in larger airports. PMID- 17915596 TI - Effects of information source, pedigree, and reliability on operator interaction with decision support systems. AB - OBJECTIVE: Two experiments are described that examined operators' perceptions of decision aids. BACKGROUND: Research has suggested certain biases against automation that influence human interaction with automation. We differentiated preconceived biases from post hoc biases and examined their effects on advice acceptance. METHOD: In Study 1 we examined operators' trust in and perceived reliability of humans versus automation of varying pedigree (expert vs. novice), based on written descriptions of these advisers prior to operators' interacting with these advisers. In Study 2 we examined participants' post hoc trust in, perceived reliability of, and dependence on these advisers after their objective experience of advisers' reliability (90% vs. 70%) in a luggage-screening task. RESULTS: In Study 1 measures of perceived reliability indicated that automation was perceived as more reliable than humans across pedigrees. Measures of trust indicated that automated "novices" were trusted more than human "novices"; human "experts" were trusted more than automated "experts." In Study 2, perceived reliability varied as a function of pedigree, whereas subjective trust was always higher for automation than for humans. Advice acceptance from novice automation was always higher than from novice humans. However, when advisers were 70% reliable, errors generated by expert automation led to a drop in compliance/reliance on expert automation relative to expert humans. CONCLUSION: Preconceived expectations of automation influence the use of these aids in actual tasks. APPLICATION: The results provide a reference point for deriving indices of "optimal" user interaction with decision aids and for developing frameworks of trust in decision support systems. PMID- 17915597 TI - Insertion loads and forearm muscle activity during flexible hose insertion tasks. AB - OBJECTIVE: To quantify the physical demands of hose insertion tasks in automotive assembly operations and how they are affected by method and the mechanical interference between the hose and the flange. BACKGROUND: Insertion tasks were identified by workers as physically demanding and can often lead to fatigue or losses in production attributable to pain or injury. METHODS: Six male and 6 female participants pushed a 25.4-mm flexible rubber hose onto a stationary flange during simulated insertions. Three insertion methods -- rock, straight, and twist -- were examined in the study. Muscle activity of the finger flexors was recorded to estimate grip effort during the simulated insertions. RESULTS: The twist method (114.8 N) resulted in a 26% reduction in axial force compared with the straight method (155.7 N). Average muscle activity ranged from a low of 14% maximum voluntary contraction (MVC; men, straight method) to a high of 67% MVC (women, twist method). Hose resultant forces ranged from a low of 52.2 N to a high of 461.1 N for all participants. Men exerted 6% higher resultant forces with 37% less muscle activity than women. CONCLUSION: There are situations when the 26% reduction in the axial force attributable to twisting may be helpful during an insertion, despite the fact that forearm muscle activity was highest for both male and female participants during twisting insertions. APPLICATION: The results of this study can be applied to the future design of tasks that involve the joining of two parts such as a hose and flange. PMID- 17915598 TI - Effect of head-mounted displays on posture. AB - OBJECTIVE: The aim of the present study was to determine if a wearable system based on a head-mounted display (HMD) causes users to alter their head position and adopt postures that place greater stress on the musculoskeletal system. BACKGROUND: HMDs are common output devices used with wearable computers. HMDs provide the wearer with visual information by projecting computer-generated virtual images in front of the eyes. Deviations of neck posture from a neutral upright position increase the stresses on the musculoskeletal system of the head and neck. METHOD: Seven paramedics simulated the treatment of a patient under a normal condition and when using an HMD wearable computer system. During the simulations a posture analysis was performed using the Rapid Upper Limb Assessment method. RESULTS: The postures adopted when wearing an HMD, as compared with a normal condition, scored significantly higher for the neck (z = 2.463, p < .05) and for overall body posture (left side of the body: z = 2.447, p < .05; right side of the body: z = 2.895, p < .05). CONCLUSION: Wearing an HMD can force the wearers to modify their neck posture. As such, the musculoskeletal system of the head and neck may be placed under increased levels of stress. APPLICATION: Potential users should be made aware that HMDs could dictate modifications in neck posture, which may have detrimental effects and may compound the weight effect of the HMD. PMID- 17915599 TI - Impact of order and load knowledge on trunk kinematics during repeated lifting tasks. AB - OBJECTIVE: To determine if lifting random unknown weights is more detrimental than lifting sequences of unknown weights and to investigate whether load knowledge impacts the effect of lifting random box weights. BACKGROUND: Much research has investigated lifting under known load conditions, but few studies have investigated unknown loads, especially when presented in random order. There has been some documentation of alteration in trunk mechanics when there is an overestimation of the unknown load. METHOD: Ten men and 10 women performed three lifting tasks: random unknown, random known, and same weight. A lumbar motion monitor was used to collect kinematic data, and Borg's Rating of Perceived Exertion (RPE) and a task risk rating were also assessed. RESULTS: Both presentation order and load knowledge impacted trunk kinematics during repeated lifting tasks. However, these differences were relatively low in magnitude. Furthermore, kinematic response and perceived risk and exertion for these conditions varied between genders. CONCLUSION: Lifting random unknown loads appears to alter kinematic responses, particularly for men. Women attempt to modify the effect of random unknown loads by changing the lifting style through alterations in upper limb motions (e.g., drag box toward them prior to lifting). However, a need remains for a more comprehensive biomechanical investigation (e.g., spine loading) into the effects of random unknown loads because many of the effect sizes were small. APPLICATION: Small kinematic adaptations resulting from tasks involving unknown and random loads may be mediated by the use of visual cues, order of presentation, or a change in lifting style. PMID- 17915600 TI - Neuromuscular response to cyclic lumbar twisting. AB - OBJECTIVE: To study the influence of 10 min of cyclic twisting motion on abdominal and back muscle activities. BACKGROUND: Repetitive (cyclic) occupational activity was identified by many epidemiological reports to be a risk factor for the development of work-related musculoskeletal disorders. Biomechanical and physiological confirmation, however, is lacking. METHODS: Trunk muscle electromyography (EMG) was recorded while participants performed a continuous 10-min maximum lumbar cyclic twisting to the left, and maximum isometric twist to the left and right sides was measured before and after the exercise. RESULTS: Abdominal muscles contracted symmetrically, independent of twisting direction. The left posterior muscles' integrated EMG (IEMG) decreased during the exercise, whereas the IEMG of the right posterior muscle increased. Simultaneously with increased antagonist coactivity level of the right posterior muscles after the exercise, decrease in maximal isometric left twisting torque was observed. The abdominal muscles did not exhibit any significant changes during the exercise. After the exercise, the right abdominals demonstrated a significant increase in effort, which was independent of the direction of the maximal effort isometric test. CONCLUSIONS: The change in muscle activity is attributed to neuromuscular compensation for the development of laxity and microdamage in the soft tissue (ligaments, discs, facet capsules, etc.) of the lumbar spine. APPLICATION: The results of this study increase understanding of the risk factors associated with low back disorder induced by labor-intensive occupations that involve cyclic lateral twisting. PMID- 17915601 TI - The effects of visual display distance on eye accommodation, head posture, and vision and neck symptoms. AB - OBJECTIVE: Determine the effects of display viewing distance on both the visual and musculoskeletal systems while the text height is held constant across viewing distances. BACKGROUND: The distance from the eyes to a computer display may affect visual and neck comfort. If the angular size of the characters remains the same, it is recommended that the display be placed at a farther viewing distance (e.g., 70-100 cm). However, in common usage, the character sizes are not adjusted based on viewing distance. METHOD: Participants under the age of 35 years (N = 24) performed visually demanding tasks using a computer display for 2 hr each at three viewing distances (mean: 52.4, 73.0, and 85.3 cm) while torso and head posture were tracked. At the end of each task, eye accommodation was measured and symptoms were recorded. RESULTS: The near distance was associated with significantly less blurred vision, less dry or irritated eyes, less headache, and improved convergence recovery when compared with the middle and far distances. Participants moved their torsos and heads closer to the monitor at the far distance. CONCLUSION: If the computer screen character sizes are close to the limits of visual acuity, it is recommended that the computer monitor be positioned between the near (52 cm) and middle (73 cm) distance from the eyes. APPLICATION: The location of a computer display should take into account the size of the characters on the screen and the visual acuity of the user. PMID- 17915603 TI - A meta-analysis of performance response under thermal stressors. AB - OBJECTIVE: Quantify the effect of thermal stressors on human performance. BACKGROUND: Most reviews of the effect of environmental stressors on human performance are qualitative. A quantitative review provides a stronger aid in advancing theory and practice. METHOD: Meta-analytic methods were applied to the available literature on thermal stressors and performance. A total of 291 references were collected. Forty-nine publications met the selection criteria, providing 528 effect sizes for analysis. RESULTS: Analyses confirmed a substantial negative effect on performance associated with thermal stressors. The overall effect size for heat was comparable to that for cold. Cognitive performance was least affected by thermal stressors, whereas both psychomotor and perceptual task performance were degraded to a greater degree. Other variables were identified that moderated thermal effects. CONCLUSION: Results confirmed the importance of task type, exposure duration, and stressor intensity as key variables impacting how thermal conditions affect performance. Results were consistent with the theory that stress forces the individual to allocate attentional resources to appraise and cope with the threat, which reduces the capacity to process task-relevant information. This represents a maladaptive extension of the narrowing strategy, which acts to maintain stable levels of response when stress is first encountered. APPLICATION: These quantitative estimates can be used to design thermal tolerance limits for different task types. Although results indicate the necessity for further research on a variety of potentially influential factors such as acclimatization, the current summary provides effect size estimates that should be useful in respect to protecting individuals exposed to adverse thermal conditions. PMID- 17915602 TI - Dual-task performance consequences of imperfect alerting associated with a cockpit display of traffic information. AB - OBJECTIVE: Performance consequences related to integrating an imperfect alert within a complex task domain were examined in two experiments. BACKGROUND: Cockpit displays of traffic information (CDTIs) are being designed for use in airplane cockpits as responsibility for safe separation becomes shared between pilots and controllers. Of interest in this work is how characteristics of the alarm system such as threshold, modality, and number of alert levels impact concurrent task (flight control) performance and response to potential conflicts. METHODS: Student pilots performed a tracking task analogous to flight control while simultaneously monitoring for air traffic conflicts with the aid of a CDTI alert as the threshold, modality, and level of alert was varied. RESULTS: As the alerting system became more prone to false alerts, pilot compliance decreased and concurrent performance improved. There was some evidence of auditory preemption with auditory alerts as the false alarm rate increased. Finally, there was no benefit to a three-level system over a two-level system. CONCLUSION: There is justification for increased false alarm rates, as miss-prone systems appear to be costly. The 4:1 false alarm to miss ratio employed here improved accuracy and concurrent task performance. More research needs to address the potential benefits of likelihood alerting. APPLICATION: The issues addressed in this research can be applied to any imperfect alerting system such as in aviation, driving, or air traffic control. It is crucial to understand the performance consequences of new technology and the efficacy of potential mitigating design features within the specific context desired. PMID- 17915604 TI - Optical information for car following: the driving by visual angle (DVA) model. AB - OBJECTIVE: The present study developed and tested a model of car following by human drivers. BACKGROUND: Previous models of car following are based on 3-D parameters such as lead vehicle speed and distance information, which are not directly available to a driver. In the present paper we present the driving by visual angle (DVA) model, which is based on the visual information (visual angle and rate of change of visual angle) available to the driver. METHOD: Two experiments in a driving simulator examined car-following performance in response to speed variations of a lead vehicle defined by a sum of sine wave oscillations and ramp acceleration functions. In addition, the model was applied to six driving events using real world-driving data. RESULTS: The model provided a good fit to car-following performance in the driving simulation studies as well as in real-world driving performance. A comparison with the advanced interactive microscopic simulator for urban and nonurban networks (AIMSUN) model, which is based on 3-D parameters, suggests that the DVA was more predictive of driver behavior in matching lead vehicle speed and distance headway. CONCLUSION: Car following behavior can be modeled using only visual information to the driver and can produce performance more predictive of driver performance than models based on 3-D (speed or distance) information. APPLICATION: The DVA model has applications to several traffic safety issues, including automated driving systems and traffic flow models. PMID- 17915605 TI - Temporal limitations in multiple target detection in a dynamic monitoring task. AB - OBJECTIVE: Two experiments examined the detectability of multiple transient changes within a cluttered and dynamic display (a simulated sonar display). BACKGROUND: Research suggests that there are severe limitations when multiple targets must be detected within close temporal proximity. The present research explored whether these limitations influence performance in a dynamic monitoring task. METHOD: Participants monitored a cluttered and dynamic display and reported the number of new objects that appeared (one to four objects). The time between onset events was varied. A blinking cue sometimes accompanied each new object, giving observers multiple opportunities to detect it. RESULTS: A large decrease in performance was observed when participants were asked to detect multiple targets within a short period of time. Performance was worse than predicted based on the attention literature. Performance suffered when observers were asked to detect more than two or three targets. The blinking cue greatly attenuated this performance deficit, even for short-duration blinking cues (one blink). CONCLUSION: Operators can easily become overwhelmed when asked to respond to even a small number of events when these events occur close in time. Extending transient events in time improves performance, but some attentional limitations may be difficult or impossible to overcome. APPLICATION: These results have important implications for systems in which important events may occur within close temporal proximity (e.g., when a sonar operator is tasked with detecting threats in the battle space). Situations in which these limitations may or may not influence performance are discussed. PMID- 17915606 TI - Depth of focus and visual recognition of imagery presented on simultaneously viewed displays: implications for head-mounted displays. AB - OBJECTIVE: We sought to determine the optimal focal distance for a semitransparent monocular head-mounted display (HMD) integrated with a flight simulator display and to investigate whether observers experienced visual discomfort or impaired target recognition when using an HMD set at the optimal distance. BACKGROUND: When an observer wears a monocular HMD and views a simulator display, focal distances of both displays must be within the observers' depth of focus to prevent blurred imagery. Because focal distance can vary by as much as 0.5 m in U.S. Air Force multifaceted simulator displays, we determined whether a monocular HMD could be integrated with a simulator display without blurred imagery or discomfort. METHOD: Depth of focus and visual recognition were measured with a staircase procedure, and visual discomfort was measured with a questionnaire. RESULTS: Depth of focus was 0.64 diopters in one condition tested, but it was affected by luminance level and display resolution. It was recommended that HMD focal distance equal the optical midpoint of the range of viewing distances encountered in the simulator. Moreover, wearing an HMD produced a decline in recognition performance for targets presented on the simulator display despite both displays being within observers' depth of focus and producing no visual discomfort. CONCLUSION: Monocular HMDs can be integrated with multifaceted simulator displays without blurred imagery or visual discomfort, provided that the correct focal distance is adopted. APPLICATION: For situations involving simultaneously viewed visual displays. PMID- 17915607 TI - Motion sickness, console video games, and head-mounted displays. AB - OBJECTIVE: We evaluated the nauseogenic properties of commercial console video games (i.e., games that are sold to the public) when presented through a head mounted display. BACKGROUND: Anecdotal reports suggest that motion sickness may occur among players of contemporary commercial console video games. METHODS: Participants played standard console video games using an Xbox game system. We varied the participants' posture (standing vs. sitting) and the game (two Xbox games). Participants played for up to 50 min and were asked to discontinue if they experienced any symptoms of motion sickness. RESULTS: Sickness occurred in all conditions, but it was more common during standing. During seated play there were significant differences in head motion between sick and well participants before the onset of motion sickness. CONCLUSION: The results indicate that commercial console video game systems can induce motion sickness when presented via a head-mounted display and support the hypothesis that motion sickness is preceded by instability in the control of seated posture. APPLICATION: Potential applications of this research include changes in the design of console video games and recommendations for how such systems should be used. PMID- 17915608 TI - Influence of age and proximity warning devices on collision avoidance in simulated driving. AB - OBJECTIVE: We conducted a set of experiments to examine the utility of several different uni- and multimodal collision avoidance systems (CASs) on driving performance of young and older adult drivers in a high-fidelity simulator. BACKGROUND: Although previous research has examined the efficacy of different CASs on collision avoidance, there has been a dearth of studies that have examined such devices in different driving situations with different populations of drivers. METHOD: Several different CAS warnings were examined in varying traffic and collision configurations both without (Experiment 1a) and with (Experiment 2) a distracting in-vehicle task. RESULTS: Overall, collision avoidance performance for both potential forward and side object collisions was best for an auditory/visual CAS, which alerted drivers using both modalities. Interestingly, older drivers (60-82 years of age) benefited as much as younger drivers from the CAS, and sometimes they benefited more. CONCLUSION: These data suggest that CASs can be beneficial across a number of different driving scenarios, types of collisions, and driver populations. APPLICATION: These results have important implications for the design and implementation of CASs for different driver populations and driving conditions. PMID- 17915610 TI - Time for a strategic imperative. PMID- 17915609 TI - Lateral control assistance for car drivers: a comparison of motor priming and warning systems. AB - OBJECTIVE: This paper's first objective is to determine whether motor priming assistance (consisting of directional steering wheel vibrations) can be of some benefit compared with more traditional auditory (lateralized sound) or vibratory (symmetric steering wheel oscillation) warning devices. We hypothesize that warning devices favor driving situation diagnosis, whereas motor priming can improve the initiation of action even further. Another objective is to assess the possible benefits of using multimodal information by combining auditory warning with simple steering wheel vibration or motor priming. BACKGROUND: Within the context of active safety devices, the experiment dealt with moderately intrusive driving assistance devices that intervene when a certain level of risk in terms of lane departure is reached. METHOD: An analysis of the steering behavior of 20 participants following episodes of visual occlusion was carried out. Five warning and motor priming devices were compared. RESULTS: All tested devices improved the drivers' steering performance, although their effects were modulated by the drivers' risk assessment. However, performance improvements were found to be greater with a motor priming device. No additional performance enhancement was observed when auditory warning was added to steering wheel vibration or motor priming devices. CONCLUSION: This study confirms the hypothesis that the direct intervention of motor priming at the action level is more effective than a simple warning, which intervenes upstream in situation diagnosis. Multimodal information did not seem to improve driver performance. APPLICATION: This study proposes a new kind of lateral control assistance, which acts at a sensorimotor level, in contrast with traditional warning devices. PMID- 17915611 TI - More than the blues: perinatal depression. PMID- 17915612 TI - The emerging culture of health care: from horizontal violence to true collaboration. PMID- 17915613 TI - A flat world of health care. PMID- 17915614 TI - Drug use review matters. PMID- 17915615 TI - Correlation between pulse wave velocity and other measures of arterial stiffness in chronic kidney disease. AB - AIM: Pulse wave velocity (PWV), augmentation index (AIx) and time to first wave reflection (Tr) are all measures of arterial stiffness, but whether these parameters behave similarly in different populations is not well-understood. Given the large burden of cardiovascular disease in individuals with chronic kidney disease (CKD), assessing the relationship between vascular stiffness parameters in this population is important. METHOD: A subset of 152 participants enrolled in the Chronic Renal Insufficiency Cohort Study had vascular stiffness parameters (aortic PWV, central AIx, and Tr) measured using the SphygmoCor system. Linear association between these parameters was assessed using Pearson correlation coefficients. Reproducibility across operators of the device was also tested within individuals. RESULTS: Association was largest between PWV and heart rate-adjusted AIx (AIx-75). The correlation coefficient was 0.371 (p = 0.0003) for ideal studies and 0.305 (p = 0.0001) for all technically acceptable studies. The association between ideal PWV and AIx-75 measurements was 0.361 (p = 0.005) for men and 0.423 (p = 0.01) for women. Bland-Altman plots comparing the mean value of PWV (n = 31) or AIx-75 (n = 21) when measured by 2 different individuals against the difference in their respective values demonstrate that both measures of arterial stiffness are reproducible across multiple technicians. CONCLUSIONS: Thus, we conclude that PWV and AIx-75, despite measuring different quantities in different units, are related measures of arterial stiffness and are reproducible across multiple operators in the population with CKD. PMID- 17915616 TI - Oxidative status and prevalent cardiovascular disease in patients with chronic renal failure treated by hemodialysis. AB - BACKGROUND: Hemodialysis (HD) patients are exposed to oxidative stress which contributes to cardiovascular disease (CVD). The purpose of this study was to investigate the oxidative and antioxidative status in HD patients with (CVD+, n = 38) and without (CVD-, n = 67) prevalent CVD. PATIENTS AND METHODS: A total of 105 HD patients and 21 healthy controls were assessed for lipid peroxidation indices (plasma malondialdehyde (MDA)), oxidizability of apolipoprotein B containing lipoproteins (apo B-deltaMDA) and red blood cells (RBC-MDA) together with various components of the antioxidant system in plasma (paraoxonase/arylesterase activities, total carotenoids, vitamins C and E) and RBC (superoxide dismutase and glutathione peroxidase activities). RESULTS: Plasma MDA and RBC-MDA were significantly higher, vitamin C and total carotenoid levels were significantly lower in both CVD+ and CVD- HD groups than in the control group. Plasma MDA levels were significantly higher and serum paraoxonase activity, uric acid and albumin levels were significantly lower in patients with CVD+ HD patients compared to those of the CVD- patients. CONCLUSION: This study suggests that the elevated level of plasma MDA and the lower activity of paraoxonase could contribute to the increased incidence of cardiovascular disease in hemodialysis patients. PMID- 17915617 TI - The economics of home nocturnal hemodialysis: how should we cost the benefits? AB - Home nocturnal hemodialysis (HNHD) has been established as a safe and effective way to provide dialysis for patients who require renal replacement therapy. Non randomized studies have shown that patients switched to HNHD have improvements in blood pressure, left ventricular mass and quality of life. At present, there are no RCTs or long-term observational studies demonstrating a clear reduction in cardiovascular events or mortality. Several HNHD centers have published articles documenting the costs of this modality as compared to conventional HD. Some of these studies have found HNHD to provide significant cost savings, while others have found the two modalities to be relatively equivalent in terms of costs. In this paper, we review the results of these costing studies and illustrate some of the limitations associated with these studies including the lack of randomization, inconsistent reporting of HNHD start-up costs, potential patient selection biases and limited follow-up. On balance, it appears premature to conclude that HNHD is cost-saving in comparison to conventional hemodialysis. However, two ongoing randomized trials, which are collecting resource use information, will help to answer this question. Once these data are available, a formal economic evaluation should be done to determine the impact of HNHD on both clinical outcomes and costs. This information will assist decision-makers in determining whether to make HNHD more widely available. PMID- 17915618 TI - A pilot study in hemodialysis of an electrophysiological tool to measure sudden cardiac death risk. AB - BACKGROUND: The hemodialysis procedure may play a role in the elevated risk of sudden cardiac death seen in hemodialysis patients. METHODS: Microvolt T wave alternans, a promising noninvasive electrophysiological test developed to measure sudden cardiac death risk, was used to test the hypotheses that high-risk hemodialysis patients commonly manifest cardiac electrophysiology that is associated with higher sudden death risk in nondialysis patients and that the hemodialysis procedure modifies cardiac electrophysiology in a manner predisposing to malignant ventricular arrhythmias. To test this hypothesis, microvolt T wave alternans tracings were done in 9 patients before and immediately after an early week hemodialysis session. RESULTS: 7 of 9 individuals had non-negative (i.e. higher risk) tracings either before or after hemodialysis. 2 of 4 subjects with tracings initially negative before hemodialysis became non negative after hemodialysis. CONCLUSION: This pilot study provides the first objective preliminary evidence using microvolt T wave alternans that high-risk hemodialysis patients commonly exhibit abnormal cardiac repolarization and that hemodialysis treatments can acutely alter repolarization in a potentially harmful manner. PMID- 17915619 TI - ANCA-related crescentic glomerulonephritis in systemic sclerosis: revisiting the "normotensive scleroderma renal crisis". AB - The scleroderma renal crisis is characterized by acute onset of severe hypertension and by rapidly progressive hyperreninemic renal failure. There is, however, a very limited subset of patients with rapidly progressive renal failure who remain normotensive and develop ANCA-positive crescentic glomerulonephritis. We report a case of normotensive acute renal failure secondary to anti-MPO antibody-associated crescentic glomerulonephritis in a patient with diffuse systemic sclerosis. She was referred to our department with normal blood pressure and no extrarenal clinical manifestation ofvasculitis. She presented with rapidly progressive renal failure, microscopic hematuria and minimal proteinuria. P-ANCA were positive by immunofluorescence, with ELISA-confirmed specificity for myeloperoxidase. Renal biopsy revealed typical features of pauciimmune glomerulonephritis with crescent formation and fibrinoid necrosis. The patient was initially treated with i.v. cyclophosphamide only. Because of ongoing deteriorating renal function, additional treatment with intravenous pulses of methylprednisolone followed by oral prednisone was started and allowed renal function improvement. After 9 months, serum creatinine had almost returned to normal level with minimal proteinuria, no hematuria and negative ANCA testing. Control kidney biopsy only revealed scar lesions. The association of ANCA positive crescentic glomerulonephritis and systemic sclerosis is a very rare event. Treatment with intravenous cyclophosphamide and corticosteroids allows rapid and long-term improvement of renal function. The onset of typical scleroderma renal crisis triggered by high-dose corticosteroids is unlikely but requires a close follow-up of patients with overlapping systemic sclerosis. Diagnosis and treatment are discussed and previously published cases are reviewed. PMID- 17915620 TI - Multicentric Castleman disease with secondary AA renal amyloidosis, nephrotic syndrome and chronic renal failure, remission after high-dose melphalan and autologous stem cell transplantation. AB - Multicentric Castleman disease is a systemic lymphoproliferative disease with incomplete understood etiology. The various renal complications of this disease may include minimal change disease, mesangial proliferative glomerulonephritis, membranous glomerulonephritis and nephrotic syndrome, caused by secondary amyloidosis. In several reported cases of localized Castleman disease associated with renal amyloidosis and nephrotic syndrome, resection of organs involved by lymphoid proliferation resulted in complete remission. However, therapy of multicentric Castleman disease with renal amyloidosis is not well-established. We treated a case of a 39-year-old woman with multicentric Castleman disease complicated by nephrotic syndrome caused by secondary AA amyloidosis. The patient underwent autologous peripheral blood stem cell transplantation (auto-PBSCT), achieving complete remission. Autologous stem cell transplantation may be an attractive choice in therapy for refractory multicentric Castleman disease. PMID- 17915621 TI - Ibuprofen-induced HUS. AB - BACKGROUND: Hemolytic uremic syndrome (HUS) is a disease characterized by nonimmune hemolytic anemia, thrombocytopenia and renal impairment. There are many causes for HUS, but adverse reactions to drugs have been increasingly reported. Even the NSAIDs which have been reported as safe and effective painkillers are described as cause of recurrent HUS. PATIENT CASE: We describe a case of a 44 year-old woman who was admitted to our hospital because of thrombocytopenia and anemia after the use of 8 tablets of 400 mg ibuprofen (NSAIDs). The diagnosis HUS was made and she recovered completely after treatment with fresh-frozen plasma and seven plasma exchanges. CONCLUSION: No cause could be identified except the use of ibuprofen. Recognition of a drug-induced HUS is necessary to avoid reexposure and recurrent HUS. PMID- 17915622 TI - Early detection and successful treatment of cerebral venous thrombosis associated with minimal change nephrotic syndrome. AB - Although venous thrombosis is a major complication in nephrotic syndrome, cerebral venous thrombosis (CVT) is rarely reported. We describe a 29-year-old male with nephrotic syndrome who suddenly developed headache and nausea. Although computed tomography scan and magnetic resonance imaging detected no abnormal lesions, phase-contrast magnetic resonance venography (PC MRV) demonstrated extensive thrombosis of the superior sagittal sinus. After receiving systemic anticoagulant therapy and oral prednisolone, his neurological symptoms improved dramatically, and complete remission from nephrotic syndrome was achieved. Follow up PC MRV demonstrated recanalization of the superior sagittal sinus, and renal biopsy confirmed the diagnosis of minimal change nephrotic syndrome. Although CVT is difficult to detect with conventional diagnostic methods, PC MRV may help the establishment of an early diagnosis and prompt treatment for a successful outcome. PMID- 17915623 TI - Goodpasture's syndrome in a patient using cocaine--a case report and review of the literature. AB - Glomerulonephritis is a very rare form of cocaine-induced renal pathology. We report a 26-year-old woman having inhaled cocaine, who presented with antiglomerular basement membrane glomerulonephritis and pulmonary hemorrhage. She was treated with immunosuppressive therapy and plasmapheresis. Maintenance hemodialysis was required on discharge. We stress the importance of early detection and treatment of this renal association for the potential fatal consequences implicated. PMID- 17915624 TI - Tumor lysis syndrome and acute renal failure--an increasing spectrum of presentations. AB - Tumor lysis syndrome has historically been associated with hyperuricemia and uric acid crystal deposition. We present three cases of tumor lysis syndrome resulting in renal failure in the context of normouricema, highlighting the spectrum of clinical presentations and mechanisms of renal damage. Two cases occurred following the treatment of hematological malignancies and were associated with hyperphosphatemia; the third resulted from ischemic necrosis following transarterial chemoembolization of a hepatic tumor. We also discuss the role of renal biopsy in the investigation of tumor lysis syndrome. PMID- 17915625 TI - Roundup intoxication and a rationale for treatment. AB - A 51-year-old man with no history ofrenal disease was admitted to our hospital after an intentional ingestion of Roundup, a glyphosate-based herbicide. His course was significant for the development of acute renal failure with oliguria and severe hypoxia. Although efficacy data are sparse and controversial, we proceeded with hemodialysis in an effort to correct his worsening volume status and to potentially clear toxins that are normally excreted by the kidney. His condition improved immediately and his renal function returned to normal over the course of several weeks. We argue that hemodialysis in the setting of such herbicide ingestions may facilitate significant intoxicant clearance, especially in the setting of impaired glomerular filtration. We also make recommendations regarding possible toxin-related sequelae that may warrant initiation of hemodialysis therapy. PMID- 17915626 TI - Celiac sprue-associated membranous nephropathy. PMID- 17915627 TI - Long-term mizoribine intermittent pulse therapy, but not azathioprine therapy, attenuated histologic progression in a patient with severe lupus nephritis. PMID- 17915628 TI - Integrating knowledge-based resources into the electronic health record: history, current status, and role of librarians. AB - Satisfying clinical information needs remains a major challenge in medicine, underscored by recent studies showing high medical error rates and suboptimal physician adherence to evidence-based practice guidelines. Advanced clinical decision support systems can improve practitioner performance and patient outcomes. Similarly, integrating online information resources into electronic health records (EHRs) shows great potential for positively impacting health care quality. This paper explores the evolution and current status of knowledge-based resource linkages within EHRs, including the benefits and drawbacks, as well as the important role librarians can play in this process. PMID- 17915629 TI - Environmental health and toxicology resources of the United States National Library of Medicine. AB - For over 40 years, the National Library of Medicine's (NLM) Toxicology and Environmental Health Information Program (TEHIP) has worked to organize and to provide access to an extensive array of environmental health and toxicology resources. During these years, the TEHIP program has evolved from a handful of databases developed primarily for researchers to a broad range of products and services that also serve industry, students, and the general public. TEHIP's resources include TOXNET , a collection of databases, including online handbooks, bibliographic references, information on the release of chemicals in the environment, and a chemical dictionary. TEHIP also produces several resources aimed towards the general public, such as the Household Products Database , which helps users explore chemicals often found in common household products, and Tox Town , an interactive guide to commonly encountered toxic substances, health, and the environment. This paper introduces some of NLM's environmental health and toxicology resources. PMID- 17915630 TI - The hospital library as a "magnet force" for a research and evidence-based nursing culture: A case study of two magnet hospitals in one health system. AB - When Baptist Hospital of Miami, then South Miami Hospital, became Magnet award winning hospitals, their libraries' challenges increased. Could their librarians ease the transition of research and evidence-based practice into the "real world" of nursing? Did library services have a role in the ongoing Magnet re credentialing process? This case study defines hospital library magnet force strategies that worked in the quest for this prestigious award for nursing excellence at two hospitals at Baptist Health South Florida. PMID- 17915631 TI - Using MeSH to search for alternatives to the use of animals in research. AB - Searching for alternatives to using animals in research is not a standard service currently offered by most medical research libraries. The goal of this article is to demystify this type of expert search for medical librarians and to do so using a language they know well, that of the Medical Subject Headings (MeSH) thesaurus. An attempt is made in this paper to discuss possible search strategies and to include examples of recommended approaches to searching-all in the context of the 3Rs of alternatives: Replacement, Refinement, and Reduction. PMID- 17915632 TI - HazLit: a unique resource for natural hazards information. AB - The "Online Updates" column generally focuses on bibliographic databases that may be licensed from commercial vendors or are available free of charge from a federal government agency. HazLit is featured because it is a unique and comprehensive resource of the literature of natural disasters. This column provides background information on HazLit and offers some searching basics, as well as highlights special information found at the Web site that hosts the database. PMID- 17915633 TI - Evolution of the Web revolution. AB - The "Navigate the Net" column began in 1994 as a vehicle for health sciences librarians to share their explorations in finding Web-related resources located on the Internet. Over the years, there have been tremendous changes and growth in this very important information vehicle. The current editors of the column assumed their responsibility for the column in the spring of 1998. This article serves as a brief overview reflecting the changes in both the Internet and the resources hosted, maintained, and scrutinized by health sciences librarians and information professionals. PMID- 17915634 TI - Library outreach near and far: programs to staff and patients of the Piedmont Healthcare System. AB - The librarians at the Sauls Memorial Library at Piedmont Hospital describe two services they offer to Piedmont Healthcare personnel and patients. Included are the system-wide library outreach program and the Sauls Memorial Library Business Center at the main hospital. These services create good will between patients, employees, and the healthcare system, and are also vital elements in promoting and marketing the library. This article illustrates the steps that were taken to create these services within Piedmont Healthcare's growing system. PMID- 17915635 TI - Technological tools for library user education: one library's experience. AB - In today's world, library users are confronted with almost too many options for using information because of the ubiquitousness of technology. Yet, libraries can harness the power of the same technologies to help users find the information they need at the time it is needed. The tools described in this article represent a starting point for librarians looking for technologies that are easy to use, inexpensive, and have a reasonable learning curve. Technologies addressed include classroom technologies such as audience response systems and Web-based technologies, including Web tutorials and screencasting. These technologies enhance and offer flexibility and variety in many educational settings. PMID- 17915636 TI - Biopharmaceutical characterization of carbamazepine immediate release tablets. In vitro-in vivo comparison. AB - A growing concern for the biopharmaceutical characterization of pharmaceutical products increased the interest in the evaluation and identification of physicochemical properties of drugs and dosage forms that govern its biological performance. In vitro and in vivo characteristics of two carbamazepine (CAS 298 46-4) immediate release tablets were investigated and compared in order to establish level A in vitro-in vivo correlation. An in vivo study was conducted as a controlled, two-way, complete cross-over, single dose, pharmacokinetic trial in 18 subjects. The in vitro study was performed using various dissolution media in order to evaluate their potential influence on drug release and distinguish the set of experimental conditions relevant to the in vivo behavior of the investigated drug products. Beside significant differences among in vitro release profiles, the in vivo data indicated bioequivalence of the two formulations. Although a high level of correlation between in vivo and in vitro data was observed in some media, there was no single in vitro-in vivo correlation model applicable to both investigated products. The obtained results add to the existing debate on the rationale for the use of surfactants in drug release media and their in vivo relevance, emphasizing the importance of in vitro dissolution testing in addition to in vivo bioequivalence testing. PMID- 17915637 TI - Comparison of plasma and saliva concentrations of lamotrigine in healthy volunteers. AB - The relationship between lamotrigine (CAS 84057-84-1) concentrations in saliva and plasma in healthy volunteers were examined, as well as the possibility of using saliva to monitor levels for effective therapy. The study was performed with 14 healthy volunteers, mean age 23 +/- 2 (SD) years. After single oral doses of 200 mg, plasma and stimulated saliva samples were collected simultaneously at 0, 0.25, 0.5, 1, 2, 4, 6, 8, 10, 12, 24, 48, 72 and 96 h. The pH values of saliva samples were recorded. Lamotrigine concentrations were determined by a validated HPLC method. Fraction of drug bound to plasma proteins was calculated mathematically by the modified Henderson-Hasselbalch equation. Linear regression was used to evaluate the correlations. The remnant of orally administered drug contaminated the saliva samples and gave spuriously high values for up to 2 h, which were omitted. There was significant correlation (r2 = 0.677, p < 0.0001) between plasma and saliva concentrations from 2-96 h after administration. The mean ratio of saliva to plasma concentration was 0.426 +/- 0.153 (mean +/- SD). Protein binding, calculated from the concentrations in saliva was 57.5 +/-15.1% (mean +/- SD). Noncompartmental analysis was conducted with the program Kinetica. Plasma t1/2 and MRT were not significantly different from those found from saliva. The mean values of lamotrigine peak saliva concentrations (C(max)), areas under the curve of concentration versus time from zero to infinity (AUC(0- >infinity)), and areas under the curves of the product of time and concentration versus time from zero to infinity (AUMC(0-->infinity) were proportionally lower than in plasma. The results support the use of saliva concentration as a convenient, painless and noninvasive alternative to plasma for monitoring lamotrigine therapy. PMID- 17915638 TI - Bioequivalence of two tablet formulations of helicidum adminstered in single dose to healthy Chinese volunteers. AB - In this 2 x 2, randomized, crossover bioequivalence study, two tablet preparations of helicidum were compared in 20 healthy Chinese male subjects. The drug was given in a single dose of three tablets (75 mg) and blood samples were withdrawn during 12 h after drug administration. Helicidum was separated and analyzed using a validated liquid chromatography-mass spectrum method. The pharmacokinetic parameters were determined from the plasma concentration-time profiles of both formulations. The primary calculated pharmacokinetic parameters were compared statistically to evaluate bioequivalence between the two preparations, using various statistical methods. The analysis of variance (ANOVA) did not show any significant difference between the two formulations and 90% confidence intervals fell within the acceptable range (80-120%) for bioequivalence. Based on these statistical inferences it can be concluded that the two tablet preparations of helicidum are likely to be bioequivalent. PMID- 17915639 TI - Effect of benidipine hydrochloride, a long-acting T-type calcium channel blocker, on blood pressure and renal function in hypertensive patients with diabetes mellitus. Analysis after switching from cilnidipine to benidipine. AB - BACKGROUND: Calcium channel blockers are commonly used to treat hypertension, and are known to generally act on the L-type calcium channel. Recent studies have shown, however, that some calcium channel blockers also block other calcium channel subtypes, including N- and T-type channels. Cilnidipine (CAS 132203-70-4) is an L- and N-type calcium channel blocker, and benidipine hydrochloride (benidipine, CAS 91599-74-5) is known to inhibit the T-type as well as L- and N type calcium channels. In this study, effects of switching from cilnidipine to benidipine on blood pressure (BP) lowering and renal functions were investigated in order to clarify the physiological properties of the T-type calcium channel. METHODS AND RESULTS: Forty hypertensive patients with diabetes and poor BP control despite receiving cilnidipine were selected, and the changes in BP and urine protein (UP) scores were investigated retrospectively after switching from cilnidipine to benidipine for more than 3 months. BP (systolic/diastolic) significantly decreased from 155.8 +/- 13.7 mmHg/76.5 +/- 13.3 mmHg to 145.9 +/- 17.0 mmHg/71.4 +/- 13.7 mmHg after benidipine treatment, and this effect was stably maintained for one year. UP also significantly decreased from 1.29 to 0.67 in the mean score. The decrease in UP may be explained by a mechanism other than BP lowering effect. CONCLUSION: These results demonstrate that benidipine has a more potent antihypertensive effect than cilnidipine and also a renoprotective effect, indicating the high usefulness of benidipine in hypertensive patients with diabetes. T-type calcium channel blockade was suggested to be possibly involved in the enoprotective effect of benidipine. PMID- 17915640 TI - Synthesis and antidiabetic activity of some new chromonyl-2,4-thiazolidinediones. AB - A series of chromonyl-2,4-thiazolidinediones (VIa-f) and chromonyl-2,4 imidazolidinediones (VIIa-f) was prepared by Knoevenagel reaction of substituted benzyl-2,4-thiazolidinediones (IVa-f) and substituted benzyl-2,4 imidazolidinediones (Va-f) with chromone-3-carboxaldehyde (I). The prepared compounds were tested for their insulinotropic activities in INS-1 cells. Compounds VIa, VIb, VId and VIIe (at lower concentration; 1 microg/ml) were able to increase insulin release in the presence of 5.6 mmol/l glucose; their effects were lower than that of glibenclamide (CAS 10238-21-8). The activity of the most potent compound (VId) was still 9% less than that of glibenclamide. PMID- 17915641 TI - Quantitation of nimesulide in human plasma by high-performance liquid chromatography with ultraviolet absorbance detection and its application to a bioequivalence study. AB - A rapid and simple high-performance liquid chromatography-reversed phase (HPLC RV) method with ultraviolet detection (258 nm) was developed and validated for the quantitation of nimesulide (CAS 51803-78-2) in human plasma. After the plasma samples were extracted with 6.0 ml of dichloromethane containing the internal standard phenacetin 2 microg/ml in methanol, the analysis of the nimesulide level in the plasma samples was carried out using a reverse phase Supelcosil LC-18 (15 cm x 4.6 mm x 5 pm) column. The chromatographic separation was accomplished with an isocratic mobile phase consisting of a mixture of methanolphosphate buffer (pH 3.5; 0.01 mol/l) (55:45, v/v). The inter-assay accuracy ranged from 103.4 to 113.2%, while intra-day ranged from 105.6 to 117.5%. The inter-assay precision ranged from 11.7 to 14.6%, while intra-assay ranged from 3.2 to 9.5%. The recovery of nimesulide was determined as part of the assay validation process and was excellent. The linearity of the nimesulide curves ranged from 0.20 to 15.0 microg/ml (y = 0.3857-0.0081, r = 0.9975). Short-term stability showed that nimesulide is stable in plasma for at least 24 h at room temperature, while long term stability studies showed that nimesulide is stable in plasma for at least 180 days when stored at -20 degrees C. This validated method was successfully applied to the bioequivalence study of nimesulide in tablets in healthy volunteers. PMID- 17915642 TI - Pharmacokinetics and bioequivalence study of two brands of loxoprofen tablets in healthy volunteers. AB - The aims of this study were to assess the pharmacokinetics and bioequivalence of two brands of loxoprofen (CAS 80832-23-6) 60 mg tablets in healthy male volunteers. The several pharmacokinetic parameters were evaluated after an oral administration after an overnight fast according to a single dose, two-sequence, and cross-over randomized design with a 1-week washout interval. Serial blood samples were collected throughout 10 h after administration of the reference and test drug. Plasma was analyzed by validated HPLC with UV detection. Several pharmacokinetic parameters, including AUC(infnity), AUC(t), C(max), T(max), T1/2, and Ke were determined from blood concentrations of both formulations. AUC(t), AUC(infinity) and C(max) were evaluated for bioequivalence after log transformation of data using ANOVA with 90% confidence interval level. The parametric 90% confidence intervals of AUC(t), AUC(infinity), and C(max) were 90.13-106.34%, 91.43-106.94%, and 91.17-108.53%, respectively. All of the tested parameters were within the acceptable range of 80-125%. Based on these statistical considerations, it was concluded that the test drug was bioequivalent to the reference drug. PMID- 17915643 TI - Synthesis and evaluation of some novel benzothiazole derivatives as potential anticancer and antimicrobial agents. AB - Intensive efforts are underway worldwide to develop anticancer and antimicrobial agents. Therefore, a novel series of pyrido[2,1-b]benzo [d]thiazole and 2-(benzo [d] thiazol-2-ylamino)pyrimidine derivatives was synthesized. The synthesized compounds were evaluated for their anticancer activity. Among the tested compounds, compounds 3a, 3b, and 7 exhibited more cytotoxic action than the control drug doxorubicin (CAS 23214-92-8; IC50: 52 microg/ml). Compound 7 was the most potent cytotoxic, with an IC50 of 42.55 pg/ ml, while compounds 3a and 3b induced high cytotoxic action with IC50 values of 50.15 pg/ml and 50.45 pg/ml, respectively. Moreover, the synthesized compounds were screened for their antibacterial and antifungal activities. Compound 6 exhibited moderate antifungal activity against C. albicans with a minimum inhibitory concentration of 125 pg/ml. PMID- 17915644 TI - Synthesis of 3-substituted-5-(4-carb oxycyclohexylmethyl) - tetrahydro-2H-1,3,5 thiadiazine-2-thione derivatives as antifibrinolytic and antimicrobial agents. AB - A series of 3-substituted-5-(4-carboxycyclohexylmethyl)-tetrahydro-2H 1,3,5thiadiazine-2-thione derivatives was prepared and examined for antifibrinolytic and antimicrobial activities. Their structures were elucidated by spectral methods. Antifibrinolytic activities of these compounds, were investigated in vitro and compared to tranexamic acid (CAS 1197-18-8). Among the synthesized compounds, 3-methyl-5-(4-carboxycyclohexylmethyl)-tetrahydro-2H-1,3,5 thiadiazine-2-thione (Ia) was the most prominent one (104%) when compared to tranexamic acid. Besides, 3-ethyl-5-(4-carboxycyclohexyl-methyl)-tetrahydro-2H 1,3,5-thiadiazine-2-thione (Ib), 3-iso-propyl-5-(4-carboxycyclohexylmethyl) tetrahydro-2H-1,3,5-thiadiazine-2-thione (Id) and 3-isobutyl-5-(4 carboxycyclohexyl-methyl)-tetrahydro-2H-1,3,5-thiadiazine-2-thione (Ig) showed antifibrinolytic activity similar to tranexamic acid. Antibacterial activities of these compounds against Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis), Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa) and yeast-like fungi (Candida albicans, Candida tropicalis) were investigated by the micro-dilution method and compared with the activity of tranexamic acid, ofloxacin and fluconazole. By this way their minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC) and minimal fungicidal concentration (MFC) values were determined. Compound Ia exhibited almost equally potent activity against B. subtilis (MIC and MBC: 6.25 microg/mL). Compounds Ib Id, If-Ig and In exhibited similar bactericidal activity against B. subtilis (MBC: 12.5 microg/mL). Compounds Ik and Im showed bacteriostatic activity against S. aureus. None of the compounds exhibited activity against Gram-negative bacteria. On the other hand, all compounds had potent antifungal activities against the yeast utilized. Among the synthesized compounds, 3-methyl-5-(4 carboxycyclohexylmethyl)-tetrahydro-2H-1,3,5-thiadiazine-2-thione (Ia) seems to be the most effective compound with antifibrinolytic and antimicrobial activity. PMID- 17915645 TI - Little Hans: a centennial review and reconsideration. AB - Freud's "Analysis of a Phobia in a Five-year-old Boy" has stimulated interminable "reanalysis." The case of Little Hans, an unprecedented experimental child analytic treatment, is reexamined in the light of newer theory and newly derestricted documents. The understanding of the complex overdetermination of Hans's phobia was not possible in the heroic age of psychoanalysis. Current analytic thought, as well as distance de-idealization vis-a-vis the pioneering past, has potentiated a reformation of the case. The severe disturbance of his mother had an adverse impact on Little Hans and his family. Her abuse of Hans's infant sister has been overlooked by generations of analysts. Trauma, child abuse, parental strife, and the preoedipal mother-child relationship emerge as important issues that intensified Hans's pathogenic oedipal conflicts and trauma. With limited, yet remarkable help from his father and Freud, Little Hans nevertheless had the ego strength and resilience to resolve his phobia, resume progressive development, and forge a successful creative career. PMID- 17915646 TI - Little Hans "analyzed" in the twenty-first century. AB - Freud's monograph on the analysis of Little Hans is examined from a perspective aimed at highlighting elements of current thinking that would be considered mutative from those originally emphasized at the time it was written, and with a specific focus on the relative importance of verbal versus nonverbal interventions. PMID- 17915647 TI - Trauma and abuse in the case of Little Hans: a contemporary perspective. AB - Newly available interviews with Max and Herbert Graf describe the severe pathology of Little Hans's mother and her mistreatment of her husband and her daughter, who committed suicide as an adult. Reread in this context, the text of "A Phobia in a Five-year-old Boy" provides ample evidence of Frau Graf's sexual seduction and emotional manipulation of her son, which exacerbated his age expectable castration and separation anxiety, and her beating of her infant daughter. The boy's phobic symptoms can therefore be deconstructed not only as the expression of oedipal fantasy, but as a communication of the traumatic abuse occurring in the home. Through subliminal, indeed unconscious, injunctions conveyed in abusive behavior, parents can confirm the child's worst imaginings and immature views of the world and thereby render the child's oedipal conflicts and fantasies pathogenic. PMID- 17915648 TI - Little Hans and Freud's self-analysis: a biographical view of clinical theory in the making. AB - For nearly a century, Freud's "Analysis of a Phobia in a Five-year-old Boy" has been read mainly--if often critically--with Freud's conscious aim in mind: providing evidence for the central importance of oedipal conflict. Material recently released by the Freud Archives casts new light on Freud's treatment of Hans's mother, Olga Graf (nee Honig)--which began at the height of his self analysis in 1897--and of Hans himself. Read in the enriched context of new information from Eissler's interviews with Max Graf and Herbert Graf, two texts- Freud's 1897 letters to Fliess and the 1909 case history--illuminate possible personal motives for Freud's insistence on the primacy of oedipal conflict. PMID- 17915649 TI - Attachment and sibling rivalry in Little Hans: the fantasy of the two giraffes revisited. AB - Freud's interpretation of Little Hans's "phantasy of the two giraffes" is pivotal to his oedipal analysis that Hans has inchoate desires for sexual intercourse with his mother. Bowlby argued that Freud's focus on his oedipal theory led him to ignore preoedipal attachment-related factors that have equal plausibility in explaining the clinical data. However, Bowlby did not attempt to apply the attachment perspective to the interpretation of Hans's fantasies that form the core of the case material. A microanalysis of Hans's giraffe fantasy and the evidence used to support Freud's claims about it yields an attachment-based sibling rivalry account arguably of greater explanatory power than the oedipal account. Consistent with Bowlby's hypothesis, the evidence suggests that Hans's giraffe fantasy is about the sibling rivalry triangle involved in caregiver attachment access, rather than (or in addition to) the oedipal triangle. The issue of multiple levels of meaning and the methodological challenges raised by multiple determination is also considered. The giraffe fantasy's attachment theoretic explanation encourages a rethinking of this classic case and strengthens Bowlby's claim that the case is fruitfully viewed from an attachment perspective. PMID- 17915650 TI - Contributions to understanding therapeutic change: now we have a playground. AB - The understanding of therapeutic change is explored in two ways. The first is by providing a model of change that emphasizes moment-to-moment, "local-level" interactions in the analytic dyad. The second is to offer detailed clinical information--taken from the videotape of a child analyst's first session with a three-year-old girl--that illustrates how this change model can be useful to clinicians. The clinical material is presented in the form of verbal trascripts and descriptions of nonverbal communicative exchanges between child and analyst. Both the model of change and the technique of videotape microanalysis build upon work by infant researchers and thus show how advances in related fields can supplement traditional psychoanalytic methods. PMID- 17915651 TI - The anatomy of a symptom: concept development and symptom formation in a four year-old boy. AB - The case of a four-year-old boy with a postural symptom that resolved rapidly in the course of play therapy is presented. Various unconscious fantasies appeared to underlie the symptom. In particular, this case illustrates a young child's sophisticated capacity to abstract a complex relational feature from a set of unconscious fantasies that then became the basis of his symptom. The structure of the boy's symptom is relevant to (1) the question of what constitutes a symptom, (2) the relationship between concept development and symptom formation, and (3) the status of certain primary process mechanisms as they relate to concept development. Proposals are presented to help situate the contributions of psychoanalytic theory with respect to the domain of cognitive psychology, and to illustrate the unique contributions of each domain toward their mutual enrichment. PMID- 17915652 TI - Psychoanalytic approaches to work with children with severe developmental and biological disorders. Panel report. PMID- 17915653 TI - Play in the psychoanalytic situation. Panel report. PMID- 17915654 TI - Psychoanalytic education needs to change: what's feasible? Introduction to Wallerstein. PMID- 17915655 TI - The optimal structure for psychoanalytic education today: a feasible proposal? AB - Our psychoanalytic training system, close to a century old, has been subjected to increasing criticism, starting shortly after its creation, for failing to properly fulfill its avowed purposes. The most intense critiques have centered around the authoritarian power lodged in a self-selected training analyst elite, the inadequate development of a psychoanalytic research tradition, and the isolation of our educational structure from cognate disciplines concerned with human mental life, owing to its private and part-time nature, apart from the university with its spectrum of biological and human sciences. Efforts to reform this system, including the establishment of psychoanalytic institutes within medical school departments of psychiatry, and the further call for their autonomous placement within the university at large, with full-time students and faculty, have been only partially successful and have not become widespread. The values of the newly emerging multifaceted psychoanalytic center as the best currently achievable fundamental reform are presented. PMID- 17915656 TI - Emptiness in agoraphobia patients. AB - In light of new research findings about the efficacy of psychodynamic treatment for panic disorder and agoraphobia, it seems a prudent time to carefully address psychoanalytic thinking about the treatment of agoraphobia. The literature has highlighted oedipal contributions to its genesis and clinical unraveling in psychoanalysis. While those contributions are indeed central to the disorder, structural deficits in the self-representation often become a central focus of treatment once symptomatic remission has been achieved in psychoanalytic treatment. This aspect of the clinical presentation of agoraphobia has not yet been specifically addressed in the psychiatric literature. Some aspects of the phenomenon have been described by psychoanalysts. It is more difficult to treat this "emptiness" than the overt symptoms of agoraphobia, as described in DSM-IV. Nonetheless, this phenomenon may be one of the contributors to the chronicity of the disorder. Two clinical cases illustrate these points. PMID- 17915657 TI - Telephone analysis. Panel report. PMID- 17915658 TI - Robots and simulators: the future of otolaryngology? PMID- 17915660 TI - Endoscopic view of maxillary fungal sinusitis. PMID- 17915659 TI - Encephalocele. PMID- 17915661 TI - Posthemorrhagic polyp. PMID- 17915662 TI - Odontogenic myxoma of the maxilla. PMID- 17915663 TI - Cat-scratch disease lymphadenitis. PMID- 17915664 TI - Post-traumatic ethmoid mucocele following penetrating craniofacial injury. PMID- 17915665 TI - Clinical management of a patient with a 12-year history of a balance disorder. PMID- 17915666 TI - Patulous lower esophageal sphincter. PMID- 17915667 TI - Isolated uvulitis. PMID- 17915668 TI - Unpaid balances, write-offs, and refunds. PMID- 17915669 TI - An evaluation of auditory exostoses in 621 prehistoric human skulls from coastal Brazil. AB - Auditory exostoses are bone anomalies located on the floor of the external auditory canal. They frequently develop in individuals who participate in water sports and other aquatic activities. Their etiology is probably multifactorial; development seems to be triggered by regular exposure to cold water as well as to low air temperatures and/or cold winds. The presence ofauditory exostoses has been recorded in human skull fossils that date back approximately 250,000 years. We conducted a study of auditory exostoses in 621 skulls of adult humans who had been part of a marine-dependent population that lived on the Brazilian coast between 5400 and 800 years ago. The overall frequency of exostoses was 22%, but there was a great variance among different subgroups (0 to 56%). In this article, we propose some possible explanations for this variance. We also hope that our study will stimulate multidisciplinary research aimed at deciphering the intricate bony messages contained in cryptic archaeologic remains. PMID- 17915670 TI - A study of middle cranial fossa anatomy and anatomic variations. AB - We conducted a study to establish standardized measurements of the common anatomic landmarks used during surgery via the middle cranial fossa approach. Results were based on high-resolution computed tomography (CT) images of 98 temporal bones in 54 consecutively presenting patients. Measurements were obtained with the assistance of the standard PACS (picture archiving and communication system) software. We found that the superior semicircular canal (SSC) dome was not the highest point on the temporal bone (i.e., the arcuate eminence) in 78 of the temporal bone images (79.6%). Pneumatization above the SSC and above the internal auditory canal (IAC) was found in 27 (27.6%) and 39 (39.8%) temporal bone images, respectively. The anterior wall of the external auditory canal was always anterior to the anterior wall of the IAC. The mean angles between the SSC and the posterior and anterior walls of the IAC were 42.3 degrees and 60.8 degrees, respectively. We also measured other distances, and we compared our findings with those published by others. We hope that the results of our study will help surgeons safely and rapidly locate anatomic landmarks when performing surgery via the middle cranial fossa approach. PMID- 17915671 TI - Assessment of quality of life among patients with sinonasal disease as determined by an Internet survey based on the Rhinosinusitis Disability Index. AB - We conducted a survey to measure patient quality of life with the Rhinosinusitis Disability Index adapted for the Internet. Our target population was patients who were seeking information on sinonasal disease on the World Wide Web. Usable responses were obtained from 896 patients with self-diagnosed sinonasal disease. According to the survey responses, patients with chronic rhinosinusitis were significantly more bothered by their condition than were patients with allergic rhinitis. Also, women were significantly more impaired by recurrent acute sinusitis and chronic rhinosinusitis than were men. Ourfindings are consistent with the results of previous surveys conducted by means other than the Internet. This project represents the first quality-of-life survey of an unsolicited group of individuals with sinonasal disease. We believe that use of the Internet will allow researchers to obtain larger patient populations than is now typical with traditional means of survey dissemination. PMID- 17915672 TI - Influence of long-term airflow deprivation on the dimensions of the nasal cavity: a study of laryngectomy patients using acoustic rhinometry. AB - We conducted a prospective study to investigate the long-term effect of nasal airflow deprivation on nasal dimensions after total laryngectomy. We evaluated 48 patients who had an initial diagnosis of laryngeal cancer; 6 were disqualified during follow-up, leaving us with data on 42 patients for our final analysis. Acoustic rhinometry was used to measure the minimum cross-sectional area (MCSA) and the volume of the nasal cavity on both the left and right sides before and after laryngectomy. In addition, patients underwent endoscopic nasal examinations and answered questionnaires pre- and postoperatively. At both the 1- and 2-year follow-ups, the mean MCSAs and the mean nasal volumes of both the left and right nostrils were significantly smaller than the preoperative values (p < 0.001). The endoscopic examinations revealed only a mild deterioration in the appearance of the nasal mucosa over the long term. Questionnaire responses obtained at the 2 year follow-up visit revealed that all 42 evaluable patients were experiencing a moderate degree of nasal obstruction while inhaling through the nose. Our data indicate that the dimensions of the nasal cavity appear to be substantially and permanently reduced after total laryngectomy. Our study had two important advantages over other similar studies. First, because ours was a prospective study, we were able to obtain preoperative data and use it to make postoperative comparisons of the same patients rather than using healthy controls as comparators. Second, we used acoustic rhinometry, while most other studies relied on anterior rhinoscopy or rhinomanometry, which are inferior methods ofmaking the evaluations in question. We believe that our findings represent a substantial contribution to our knowledge of the physiologic and functional alterations of the nasal cavity that occur as a result of a complete cessation of nasal airflow. PMID- 17915673 TI - Efficacy comparison of multiple-antigen subcutaneous injection immunotherapy and multiple-antigen sublingual immunotherapy. AB - We performed an observational study to determine whether allergen-specific sublingual immunotherapy (SLIT) is as effective as allergen-specific subcutaneous injection immunotherapy (SCIT). Our study population was comprised of 66 patients who had been taking SLIT. Of this group, 36 patients had switched to SLIT after having been treated with SCIT (group I), while the remaining 30 patients had received SLIT only (group II). A questionnaire was used to evaluate the results of treatment. In group I, 33 patients (92%) gave SLIT a favorable rating; 27 of these patients (75%) said it was just as effective as SCIT and 6 (17%) said it was actually superior(the remaining 3 patients [8%] said that SCIT was better). In group II, 27 of 30 patients (90%) said they had attained symptom relief with SLIT; 21 (70%) said that the relief had been very significant. Overall, 60 of the 66 patients (91%) expressed various degrees of satisfaction with SLIT We believe that our SLIT protocol, which is based on established guidelines for SCIT administration, is an effective, safe, well-tolerated, and easy-to-use regimen. Future prospective studies of larger groups are clearly indicated. PMID- 17915674 TI - Comparison of Helicobacter pylori colonization on the tonsillar surface versus tonsillar core tissue as determined by the CLO test. AB - We conducted a prospective study to determine the correlation between the presence or absence of Helicobacter pylori on the tonsillar surface and in the tonsillar core as determined by the Campylobacter-like organism (CLO) rapid urease enzyme test. Our study population was made up of 55 patients who underwent adenoidectomy, tonsillectomy, or both from December 2002 through April 2003 at Khalili Hospital in Shiraz, Iran. Of these 55 patients, 45 (82%) were positive and 10 (18%) were negative for H pylori colonization as determined by CLO testing. Analysis of samples obtained from individual patients revealed differences in H pylori colonization between tonsillar surface samples and the core tissue samples. Of 106 tonsils obtained from 53 patients who underwent adenotonsillectomy or tonsillectomy, H pylori was found on 56 tonsillar surface samples (53%) and 24 tonsillar core samples (23%); only 13 tonsils (12%) contained H pylori both on the surface and in the core. We conclude that a surface swab is neither specific nor sensitive as an indicator of the presence or absence of H. pylori colonization in tonsillar core tissue. PMID- 17915675 TI - Solitary fibrous tumor of the parapharyngeal space. AB - Solitary fibrous tumors are benign neoplasms of mesenchymal origin. They usually arise from the visceral or parietal pleura and peritoneum, although they have been found in many areas throughout the body. We report a case of solitary fibrous tumor of the parapharyngeal space. Microscopically, the tumor contained spindle cells with areas of marked hypercellularity without a definitepattern. Consistent with a benign lesion, there were few mitoses and no necrosis. The tumor cells stained strongly positive for CD34 and vimentin. At the 2-year follow up, the patient was well and free of local and/or distant disease. PMID- 17915676 TI - Prognostic significance of tumor-infiltrating lymphocytes in oropharyngeal cancer. AB - The presence of tumor-infiltrating lymphocytes has been shown to significantly improve clinical outcomes in many types of cancer. However; their effects on outcomes in patients with oropharyngeal cancer specifically have yet to be elucidated. We conducted a retrospective study in an effort to shed light on this issue. We reviewed the records of 48 consecutively presenting patients with oropharyngeal cancer; and we performed immunohistochemistry to analyze their archived paraffin-embedded tissue samples for the presence of CD3-positive tumor infiltrating lymphocytes. We also used real-time polymerase chain reaction testing to look for human papillomavirus type 16 (HPV-16) in the tumors. We found that patients with large numbers of tumor-infiltrating lymphocytes (CD3high) had a significantly lower incidence of metastasis at presentation than did those with low numbers of tumor-infiltrating lymphocytes (CD31low) (40.0 vs. 88.5%; p = 0.001), regardless of HPV status. When HPV status was taken into account, the correlation between a high CD3 count and a lower rate of metastasis was maintained in the HPV-positive patients but not in the HPV-negative patients. We also found that the CD3high patients had higher rates of overall survival and disease-free survival at 3 and 5 years than did the CD3low patients; however; these differences only approached but did not reach statistical significance. PMID- 17915677 TI - Use of AlloDerm implant to improve cosmesis after parotidectomy. AB - We evaluated the effectiveness and practicality of using AlloDerm, an acellular human dermal matrix graft, as an interpositional barrier in an attempt to improve the appearance of the surgical defect created by parotidectomy. We performed AlloDerm reconstruction in a series of 10 patients, and we found that normal contour was satisfactorily restored in all 10. We conclude that the use of an AlloDerm implant is a low-risk, practical option for repairing the surgical defect in postparotidectomy patients. PMID- 17915678 TI - Supercritical water oxidation for the destruction of toxic organic wastewaters: a review. AB - The destruction of toxic organic wastewaters from munitions demilitarization and complex industrial chemical clearly becomes an overwhelming problem if left to conventional treatment processes. Two options, incineration and supercritical water oxidation (SCWO), exist for the complete destruction of toxic organic wastewaters. Incinerator has associated problems such as very high cost and public resentment; on the other hand, SCWO has proved to be a very promising method for the treatment of many different wastewaters with extremely efficient organic waste destruction 99.99% with none of the emissions associated with incineration. In this review, the concepts of SCWO, result and present perspectives of application, and industrial status of SCWO are critically examined and discussed. PMID- 17915679 TI - Control factors of partial nitritation for landfill leachate treatment. AB - Anaerobic ammonium oxidation (ANAMMOX) technology has potential technical superiority and economical efficiency for the nitrogen removal from landfill leachate, which contains high-strength ammonium nitrogen (NH4(+)-N) and refractory organics. To complete the ANAMMOX process, a preceding partial nitritation step to produce the appropriate ratio of nitrite/ammonium is a key stage. The objective of this study was to determine the optimal conditions to acquire constant partial nitritation for landfill leachate treatment, and a bench scale fixed bed bio-film reactor was used in this study to investigate the effects of the running factors on the partial nitritation. The results showed that both the dissolved oxygen (DO) concentration and the ammonium volumetric loading rate (N(v)) had effects on the partial nitritation. In the controlling conditions with a temperature of 30 +/- 1 degrees C, N(v) of 0.2-1.0 kg NH4(+) N/(m3 x d), and DO concentration of 0.8-2.3 mg/L, the steady partial nitritation was achieved as follows: more than 94% partial nitritation efficiency (nitrite as the main product), 60%-74% NH4(+)-N removal efficiency, and NO2(-)-N/NH4(+)-N ratio (concentration ratio) of 1.0-1.4 in the effluent. The impact of temperature was related to N(v) at certain DO concentration, and the temperature range of 25 30 degrees C was suitable for treating high strength ammonium leachate. Ammonium oxidizing bacteria could be acclimated to higher FA (free ammonium) in the range of 122-224 mg/L. According to the denaturing gradient gel electrophoresis analysis result of the bio-film in the reactor, there were 25 kinds of 16S rRNA gene fragments, which indicated that abundant microbial communities existed in the bio-film, although high concentrations of ammonium and FA may inhibit the growth of the nitrite-oxidizing bacteria and other microorganisms in the reactor. PMID- 17915680 TI - Simultaneous biodegradation of nitrogen-containing aromatic compounds in a sequencing batch bioreactor. AB - Many nitrogen-containing aromatic compounds (NACs), such as nitrobenzene (NB), 4 nitrophenol (4-NP), aniline (AN), and 2,4-dinitrophenol (2,4-DNP), are environmentally hazardous, and their removal from contaminated water is one of the main challenges facing wastewater treatment plants. In this study, synthetic wastewater containing NB, 4-NP, 2,4-DNP, and AN at concentrations ranging from 50 to 180 mg/L was fed into a sequencing batch reactor (SBR). Analyses of the SBR system indicated that it simultaneously removed more than 99% of the NACs at loading rates of 0.36 kg NB/(m3 x d), 0.3 kg 4-NP/(m3 x d), 0.25 kg AN/(m3 x d), and 0.1 kg 2,4-DNP/(m3 x d). Bacterial groups of Bacteriodetes, Candidate division TM7, alpha-Proteobacteria, and beta-Proteobacteria were dominant in the clone libraries of 16S rRNA genes retrieved from the microbial communities in the SBR system. "Cycle tests" designed to alter feeding and aeration parameters of the SBR system demonstrated that the resident microbial biome of the SBR system responded rapidly to changing conditions. Consumption of O2 was concomitant with the apparent mineralization of NACs. Aromatic ring-cleaving dioxygenase activities suggested that (1) AN and NB were degraded via catechol 2,3 dioxygenase; (2) 4-NP was degraded via 1,2,4-benzentriol 1,2-dioxygenase; and (3) 2,4-DNP was degraded via an unresolved pathway. PMID- 17915681 TI - Drinking water production by ultrafiltration of Songhuajiang River with PAC adsorption. AB - In recent years, membrane ultrafiltration (UF) of surface water for drinking water treatment has become a more attractive technology worldwide as a possible alternative treatment to conventional clarification. To evaluate the performance of ultrafiltration membranes for treatment of surface water in North China, a 48 m2 low pressure hollow fiber membrane ultrafiltration pilot plant was constructed. Ultrafiltration was operated in cross-flow and with powdered activated carbon (PAC) adsorption. Turbidity was almost completely removed to less than 0.2 NTU (below Chinese standard 1 NTU). It was found that PAC addition enhanced organic matter removal. The combined process of PAC/UF allowed to 41% removal of COD(Mn), 46% removal of DOC and 57% decrease in UV254 absorbance. The elimination of particles, from average 12000/ml in the raw water to approximately 15/ml in the permeated, was observed. When PAC concentration was below 30 mg/L, backwashing could recovery the membrane flux with backwash interval/backwashing duration of 1/30. PMID- 17915682 TI - Alga-lysing bioreactor and dominant bacteria strain. AB - Alga-lysing bacteria have been paid much attention to in recent years. In this study, the alga-lysing strain P05 which was isolated from an immobilizing biosystem was immobilized by coke and elastic filler, forming two biological reactors. The removal efficiencies of algae, NH4(+)-N and organic matter using the two reactors were studied. The results showed that strain P05 was an ideal algal-lysing bacteria strain because it was easy to be immobilized by coke and elastic filler which are of cheap, low biodegradability and the simple immobilization procedure. After 7 d filming, the biological film could be formed and the reactors were used to treat the eutrophic water. These two reactors were of stability and high effect with low cost and easy operation. The optimal hydraulic retention time of each reactor was 4 h. The algae removal rates were 80.38% and 82.1% (in term of Chl-a) of coke reactor and filler reactor, respectively. And that of NH4(+)-N were 52.3% and 52.7%. The removal rates of COD(Mn) were 39.03% and 39.64%. The strain P05 was identified as Bacillus sp. by PCR amplification of the 16S rRNA gene, BLAST analysis, and comparison with sequences in the GenBank nucleotide database. PMID- 17915683 TI - Sludge granulation and efficiency of phase separator in UASB reactor treating combined industrial effluent. AB - Sludge granulation and the effect of gas-liquid-solid separator (GLSS) design on the efficiency of upflow anaerobic sludge blanket (UASB) and upflow anaerobic sludge filter (UASF) reactors, operating at HRTs ranging from 3 to 12 h were investigated. VSS/TS ratio gradually increased in both the reactors with increasing sludge age (from 0.5 to more than 0.7 for UASB reactor and 0.012 to 0.043 for UASF reactor). X-Ray diffraction analysis of the UASF sludge showed the presence of expanding clays revealing its additional absorption capability. Fuoraphyllite and albite precipitation related to excellular polymers of the microbial shell structure, showed the extended growth of microorganisms during sludge granulation. A gradual decrease (82%-69%) in COD removal with decreasing HRT was apparent in UASF reactor. In case of UASB reactor, this decrease was marginal because addition of GLSS device significantly improved (14%-20%) the overall efficiency of the UASB reactor. GLSS enhanced the efficiency of the UASB reactor by increasing the settleability of suspended particles and accelerating the coagulation of colloidal particles due to the velocity gradient. PMID- 17915684 TI - Particle size distribution and removal by a chemical-biological flocculation process. AB - The particle characterization from the influent and effluent of a chemical biological flocculation (CBF) process was studied with a laser diffraction device. Water samples from a chemically enhanced primary treatment (CEPT) process and a primary sediment tank process were also analyzed for comparison. The results showed that CBF process was not only effective for both the big size particles and small size particles removal, but also the best particle removal process in the three processes of CBF process, CEPT process, and PST process (primary sediment tanks). The results also indicated that CBF process was superior to CEPT process in the heavy metals removal. The high and non-selective removal for heavy metals might be closely related to its strong ability to eliminate small particles. Samples from different locations in CBF reactors showed that small particles were easier to aggregate into big ones and those disrupted flocs could properly flocculate again along CBF reactor because of the biological flocculation. PMID- 17915685 TI - Carbonaceous aerosols in PM10 and pollution gases in winter in Beijing. AB - An intensive observation of organic carbon (OC) and elemental carbon (EC) in PM10 and gaseous materials (SO2, CO, and O3) was conducted continuously to assess the characteristics of wintertime carbonaceous aerosols in an urban area of Beijing, China. Results showed that the averaged total carbon (TC) and PM10 concentrations in observation period are 30.2 +/- 120.4 and 172.6 +/- 198.3 microg/m3, respectively. Average OC concentration in nighttime (24.9 +/- 19.6 microg/m3) was 40% higher than that in daytime (17.7 +/- 10.9 microg/m3). Average EC concentrations in daytime (8.8 +/- 15.2 microg/m3) was close to that in nighttime (8.9 +/- 15.1 microg/m3). The OC/EC ratios in nighttime ranging from 2.4 to 2.7 are higher than that in daytime ranging from 1.9 to 2.0. The concentrations of OC, EC, PM10 were low with strong winds and high with weak winds. The OC and EC were well correlated with PM10, CO and SO2, which implies they have similar sources. OC and EC were not well correlated with 03. By considering variation of OC/EC ratios in daytime and night time, correlations between OC and O3, and meteorological condition, we speculated that OC and EC in Beijing PM10 were emitted as the primary particulate form. Emission of motor vehicle with low OC/EC ratio and coal combustion sources with high OC/EC ratio are probably the dominant sources for carbonaceous aerosols in Beijing in winter. A simple ratio method was used to estimate the relative contribution of sources to carbonaceous aerosols in Beijing PM10. Motor vehicle source accounts for 80% and 68%, while coal combustion accounts for 20% and 32% in daytime and nighttime, respectively in Beijing. Averagely, the motor vehicle and coal combustion accounted for 74% and 26%, respectively, for carbonaceous aerosols during the observation period. It points to the motor vehicle is dominant emission for carbonaceous aerosols in Beijing PM10 in winter period, which should be paid attention to control high level of PM10 in Beijing effectively. PMID- 17915686 TI - Modeling of air quality with a modified two-dimensional Eulerian model: a case study in the Pearl River Delta (PRD) region of China. AB - A modified two-dimensional Eulerian air quality model was used to simulate both the gaseous and particulate pollutant concentrations during October 21-24, 2004 in the Pearl River Delta (PRD) region, China. The most significant improvement to the model is the added capability to predict the secondary organic aerosols (SOA) concentrations because of the inclusion of the SOA formation chemistry. The meteorological input data were prepared using the CALMET meteorological model. The concentrations of aerosol-bound species such as NO3-, NH4+, SO4(2-), and SOA were calculated in the fine particle size range (< 2.5 microm). The results of the two-dimensional model were compared to the measurements at the ground level during the PRD Intensive Monitoring Campaign. Overall, there were good agreements between the measured and modeled concentrations of inorganic aerosol components and O3. Both the measured and the modeled results indicated that the maximum hourly 03 concentrations exceeded the China National Air Quality Standard. The predicted 24-h average SOA concentrations were in reasonable agreement with those predicted by the method of minimum OC/EC ratio. PMID- 17915687 TI - Air quality nearby road traffic tunnel portals: BTEX monitoring. AB - A monitoring campaign of BTEX (benzene, toluene, ethylbenzene, o- m- and p xylene) was carried out nearby two tunnel portals in the urban area of Naples with the aim to verify air quality in this kind of urban sites. Sampling was carried out using the active adsorption technique. Sampling time was 1 h. Ambient temperature and traffic flow measurements were carried out during each sampling operation. The results indicate that average benzene concentrations at both sites exceed the limit value of 10 microg/Nm3 established by the European Community (EC) (Dir. 2000/69). Concentration levels of other BTEX are relatively high as well. A correlation between BTEX concentration and two wheeler vehicle flow was observed. PMID- 17915688 TI - Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China. AB - A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, sigmaDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by sigmaHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and alpha-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD+DDE)/DDT in soils under three land usages were: paddy field > tree land > fallow land, indicating that land usage influenced the degradation of DDT in soils. Ratios of p,p'-(DDD+DDE)/DDT > 1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes. PMID- 17915689 TI - Seed selections for crystallization of calcium phosphate for phosphorus recovery. AB - Seed induces and promotes the crystallization of calcium phosphate, and acts as carrier of the recovered phosphorus (P). In order to select suitable seed for P recovery from wastewater, three seeds including Apatite (AP), Juraperle (JP) and phosphate-modified Juraperle (M-JP) were tested and compared. Batch and fixed-bed column experiments of seeded crystallization of calcium phosphate were undertaken by using synthetic wastewater with 10 mgP/L. It showed that AP had bad enduring property in the crystallization process, while JP had better performance for multiple uses, and M-JP was a hopeful seed for P recovery by crystallization of calcium phosphate. PMID- 17915690 TI - Oxidative stress and DNA damages induced by cadmium accumulation. AB - Experimental evidence shows that cadmium (Cd) could induce oxidative stress and then causes DNA damage in animal cells, however, whether such effect exists in plants is still unclear. In the present study, Vicia faba plants was exposed to 5 and 10 mg/L Cd for 4 d to investigate the distribution of Cd in plant, the metal effects on the cell lipids, antioxidative enzymes and DNA damages in leaves. Cd induced an increase in Cd concentrations in plants. An enhanced level of lipid peroxidation in leaves and an enhanced concentration of H2O2 in root tissues suggested that Cd caused oxidative stress in Vicia faba. Compared with control, Cd-induced enhancement in superoxide dismutase activity was more significant at 5 mg/L than at 10 mg/L in leaves, by contrast, catalase and peroxidase activities were significantly suppressed by Cd addition. DNA damage was detected by neutral/neutral, alkaline/neutral and alkaline/alkaline Comet assay. Increased levels of DNA damages induced by Cd occurred with reference to oxidative stress in leaves, therefore, oxidative stress induced by Cd accumulation in plants contributed to DNA damages and was likely an important mechanism of Cd phytotoxicity in Vicia faba plants. PMID- 17915691 TI - Response of soil fauna to simulated nitrogen deposition: a nursery experiment in subtropical China. AB - We studied the responses of soil fauna to a simulated nitrogen deposition in nursery experimental plots in Subtropical China. Dissolved NH4NO3 was applied to the soil by spraying twice per month for 16 months, starting in January 2003 with treatments of 0, 5, 10, 15 and 30 gN/(m2 x a). Soil fauna was sampled after 6, 9, 13 and 16 months of treatment in three soil depths (0-5 cm, 5-10 cm, 10-15 cm). Soil available N increased in correspondence with the increasing N treatment, whereas soil pH decreased. Bacterial and fungal densities were elevated by the N treatment. Soil fauna increased in the lower nitrogen treatments but decreased in the higher N treatments, which might indicate that there was a threshold around 10 gN/(m2 x a) for the stimulating effects of N addition. The N effects were dependent on the soil depth and sampling time. The data also suggested that the effects of the different N treatments were related to the level of N saturation, especially the concentration of NO3- in the soil. PMID- 17915692 TI - Contribution of additives Cu to its accumulation in pig feces: study in Beijing and Fuxin of China. AB - Massive amounts of pig manure are produced by intensive pig farm in China, and the composition of pig manure has changed much due to the use of feed additives. However, little is known about the exact Cu (copper) feed as additives or present as contaminants in pig feed and the residues in feces. One hundred and thirty seven feeds and one hundred and forty-two fecal samples from 48 pig farms were collected in Beijing and Fuxin cities in 1999 and 2005, respectively. The concentrations of Cu were in the range of 6.86-395.19 mg/kg in the feed samples, and the mean values were in the order of weaner > grower-finisher > sow's feeds. The high concentrations over EU recommendations implied that excessive levels of Cu are fed on many pig farms in Beijing and Fuxin. Cu was also present in high concentrations in feces, and concentrations were highly variable. Cu concentrations in the feces from grower-finisher and weaner pigs were significantly greater than feces of sows. The super-intensive and small-scale farms had higher levels of Cu in feces than the middle farms. Cu concentrations in pig feces were approximately 5-times greater than in pig feeds. Feed management in grower-finisher pigs on super-intensive and small-scale pig farms is needed to reduce high Cu concentrations in feces and risks to soil contamination while feces are land-applied. PMID- 17915693 TI - Determination of alkali-labile phosphoprotein phosphorus from fish plasma using the Tb(3+)-tiron complex as a fluorescence probe. AB - A sensitive method based on the fluorescence quenching effect of the Tb(3+)-Tiron complex is proposed for the determination of alkali-labile phosphoprotein phosphorus (ALP) released from fish plasma. The detection limit was 5.4 ng/ml (S/N = 2), and the relative standard deviation of the quenching effect (6 replicates) was 4.6%. The results obtained by the proposed method were in good agreement with those obtained by the colorimetric assay. The advantages of the present method are its relatively simple detection procedure, the lack of toxic organic solvents, and high sensitivity. PMID- 17915694 TI - Rapid determination of phenolic compounds in water samples by alternating-current oscillopolarographic titration. AB - A rapid, simple and sensitive method was demonstrated for the determination of phenolic compounds in water samples by alternating-current oscillopolarographic titration. With the presence of sulfuric acid, phenol could be transferred into a nitroso-compound by reacting with NaNO2. The titration end-point was obtained by the formation of a sharp cut in the oscillopolarographic with infinitesimal NaNO2 on double platinum electrodes. The results showed that phenol concentration had an excellent linear relationship over the range of 4.82 x 10(-6)-9.65 x 10(-3) mol/L, the RSD of the proposed method was lower than 1.5%, and the spiked recoveries of three real water samples were in the range of 95.6%-106.9%. PMID- 17915695 TI - Process conditions for preparing methanol from cornstalk gas. AB - The low-heat-value cornstalk gas produced in the down-flow fixed bed gasifier was tentatively used for methanol synthesis. The cornstalk gas was purified and the technical procedures such as deoxygenation, desulfurization, catalytic cracking of tar, purification and hydrogenation were studied. The catalytic experiments of methanol synthesis with cornstalk syngas were carried out in a tubular-flow integral and isothermal reactor. The effect of reaction temperature, pressure, catalyst types, catalyst particle size, syngas flow at entering end and composition of syngas was investigated. The optimum process conditions and yield of methanol from cornstalk syngas were obtained. The experimental results indicated that the proper catalyst for the synthetic reaction was C301 and the optimum catalyst size was 0.833 mm x 0.351 mm. The optimum operating temperature and pressure were found to be 235 degrees C and 5 MPa, respectively. The suitable syngas flow 0.9-1.10 mol/h at entering end was selected and the best composition of syngas were CO 10.49%, CO2 8.8%, N2 37.32%, C(n)H(m) 0.95% and H2 40.49%. The best methanol yield was 0.418 g/g cornstalk. This study provided the technical support for the industrial test of methanol production from biomass (cornstalk) gas. PMID- 17915696 TI - Approach of technical decision-making by element flow analysis and Monte-Carlo simulation of municipal solid waste stream. AB - This paper deals with the procedure and methodology which can be used to select the optimal treatment and disposal technology of municipal solid waste (MSW), and to provide practical and effective technical support to policy-making, on the basis of study on solid waste management status and development trend in China and abroad. Focusing on various treatment and disposal technologies and processes of MSW, this study established a Monte-Carlo mathematical model of cost minimization for MSW handling subjected to environmental constraints. A new method of element stream (such as C, H, O, N, S) analysis in combination with economic stream analysis of MSW was developed. By following the streams of different treatment processes consisting of various techniques from generation, separation, transfer, transport, treatment, recycling and disposal of the wastes, the element constitution as well as its economic distribution in terms of possibility functions was identified. Every technique step was evaluated economically. The Mont-Carlo method was then conducted for model calibration. Sensitivity analysis was also carried out to identify the most sensitive factors. Model calibration indicated that landfill with power generation of landfill gas was economically the optimal technology at the present stage under the condition of more than 58% of C, H, O, N, S going to landfill. Whether or not to generate electricity was the most sensitive factor. If landfilling cost increases, MSW separation treatment was recommended by screening first followed with incinerating partially and composting partially with residue landfilling. The possibility of incineration model selection as the optimal technology was affected by the city scale. For big cities and metropolitans with large MSW generation, possibility for constructing large-scale incineration facilities increases, whereas, for middle and small cities, the effectiveness of incinerating waste decreases. PMID- 17915697 TI - Metabolism of benzo[a]pyrene in peroxynitrite/Fe(III) porphyrin system. AB - The peroxynitrite/porphyrin biomimetic system was established to investigate the effects of peroxynitrite on benzo[a]pyrene (B[a]P) metabolism. Three model systems consisting of different iron porphyrins were compared, and the results showed that the peroxynitrite/T(p-Cl)PPFeCl system was the highest catalytic efficiency in the metabolism of B[a]P. We analyzed the B[a]P metabolites produced from this system by RP-HPLC method and firstly identified the formation of nitrobenzo[a]pyrenes which are the special metabolites of B[a]P induced by peroxynitrite. PMID- 17915698 TI - Photodegradation of perfluorooctanoic acid by 185 nm vacuum ultraviolet light. AB - The photodegradation of persistent and bioaccumulative perfluorooctanoic acid (PFOA) in water by 185 nm vacuum ultraviolet (VUV) light was examined to develop an effective technology to deal with PFOA pollution. PFOA degraded very slowly under irradiation of 254 nm UV light. However, 61.7% of initial PFOA was degraded by 185 nm VUV light within 2 h, and defluorination ratio reached 17.1%. Pseudo first-order-kinetics well simulated its degradation and defluorination. Besides, fluoride ion formed in water, 4 shorter-chain perfluorinated carboxylic acids (PFCAs), that is, perfluoroheptanoic acid, perfluorohexanoic acid, perfluoropentanoic acid, and perfluorobutanoic acid. These were identified as intermediates by LC-MS measurement. These PFCAs consecutively formed and further degraded with irradiation time. According to the mass balance calculation, no other byproducts were formed. It was proposed that PFCAs initially are decarboxylated by 185 nm light, and the radical thus formed reacts with water to form shorter-chain PFCA with one less CF2 unit. PMID- 17915699 TI - Phosphorus accumulation by bacteria isolated from a continuous-flow two-sludge system. AB - In this article, polyphosphate-accumulating organisms (PAOs) from a lab-scale continuous-flow two-sludge system was isolated and identified, the different phosphorus accumulation characteristics of the isolates under anoxic and aerobic conditions were investigated. Two kinds of PAOs were both found in the anoxic zones of the two-sludge system, one of them utilized only oxygen as electron acceptor, and the other one utilized either nitrate or oxygen as electron acceptor. Of the total eight isolates, five isolates were capable of utilizing both nitrate and oxygen as electron acceptors to uptake phosphorus to some extent. And three of the five isolates showed good phosphorus accumulative capacities both under anoxic or aerobic conditions, two identified as Alcaligenes and one identified as Pseudomonas. Streptococcus was observed weak anoxic phosphorus accumulation because of its weak denitrification capacity, but it showed good phosphorus accumulation capacity under aerobic conditions. One isolates identified as Enterobacteriaceae was proved to be a special species of PAOs, which could only uptake small amounts of phosphorus under anoxic conditions, although its denitrification capacity and aerobic phosphorus accumulation capacity were excellent. PMID- 17915700 TI - Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes. AB - In this study an effort has been made to use plant polyphenol oxidases; potato (Solanum tuberosum) and brinjal (Solanum melongena), for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation. PMID- 17915701 TI - Surface reaction of Bacillus cereus biomass and its biosorption for lead and copper ions. AB - In this study, the surface chemical functional groups of Bacillus cereus biomass were identified by Fourier transform infrared (FTIR) analytical technique. It had been shown that the B. cereus cells mainly contained carboxyl, hydroxyl, phosphate, amino and amide functional groups. The potentiometric titration was conducted to explain the surface acid-base properties of aqueous B. cereus biomass. The computer program FITEQL 4.0 was used to perform the model calculations. The optimization results indicated that three sites-three pKas model, which assumed the cell surface to have three distinct types of surface organic functional groups based on the IR analysis results, simulated the experimental results very well. Moreover, batch adsorption experiments were performed to investigate biosorption behavior of Cu(II) and Pb(II) ions onto the biomass. Obviously, the adsorption equilibrium data for the two ions were reasonably described by typical Langmuir isotherm. PMID- 17915702 TI - Removal of phenol by activated alumina bed in pulsed high-voltage electric field. AB - A new process for removing the pollutants in aqueous solution-activated alumina bed in pulsed high-voltage electric field was investigated for the removal of phenol under different conditions. The experimental results indicated the increase in removal rate with increasing applied voltage, increasing pH value of the solution, aeration, and adding Fe2+. The removal rate of phenol could reach 72.1% when air aeration flow rate was 1200 ml/min, and 88.2% when 0.05 mmol/L Fe2+ was added into the solution under the conditions of applied voltage 25 kV, initial phenol concentration of 5 mg/L, and initial pH value 5.5. The addition of sodium carbonate reduced the phenol removal rate. In the pulsed high-voltage electric field, local discharge occurred at the surface of activated alumina, which promoted phenol degradation in the thin water film. At the same time, the space-time distribution of gas-liquid phases was more uniform and the contact areas of the activated species generated from the discharge and the pollutant molecules were much wider due to the effect of the activated alumina bed. The synthetical effects of the pulsed high-voltage electric field and the activated alumina particles accelerated phenol degradation. PMID- 17915703 TI - Start-up performances of dry anaerobic mesophilic and thermophilic digestions of organic solid wastes. AB - Two dry anaerobic digestions of organic solid wastes were conducted for 6 weeks in a lab-scale batch experiment for investigating the start-up performances under mesophilic and thermophilic conditions. The enzymatic activities, i.e., beta glucosidase, N-alpha-benzoyl-L-argininamide (BAA)-hydrolysing protease, urease and phosphatase activities were analysed. The BAA-hydrolysing protease activity during the first 2-3 weeks was low with low pH, but was enhanced later with the pH increase. beta-Glucosidase activity showed the lowest values in weeks 1-2, and recovered with the increase of BAA-hydrolysing protease activity. Acetic acid dominated most of the total VFAs in thermophilic digestion, while propionate and butyrate dominated in mesophilic digestion. Thermophilic digestion was confirmed more feasible for achieving better performance against misbalance, especially during the start-up period in a dry anaerobic digestion process. PMID- 17915704 TI - Leaching of styrene and other aromatic compounds in drinking water from PS bottles. AB - Bottled water may not be safer, or healthier, than tap water. The present studies have proved that styrene and some other aromatic compounds leach continuously from polystyrene (PS) bottles used locally for packaging. Water sapmles in contact with PS were extracted by a preconcentration technique called as "purge and trap" and analysed by gas chromatograph-mass spectrometer (GC/MS). Eleven aromatic compounds were identified in these studies. Maximum concentration of styrene in PS bottles was 29.5 microg/L. Apart from styrene, ethyl benzene, toluene and benzene were also quantified but their concentrations were much less than WHO guide line values. All other compounds were in traces. Quality of plastic and storage time were the major factor in leaching of styrene. Concentration of styrene was increased to 69.53 microg/L after one-year storage. In Styrofoam and PS cups studies, hot water was found to be contaminated with styrene and other aromatic compounds. It was observed that temperature played a major role in the leaching of styrene monomer from Styrofoam cups. Paper cups were found to be safe for hot drinks. PMID- 17915705 TI - Artificial neural network model for identifying taxi gross emitter from remote sensing data of vehicle emission. AB - Vehicle emission has been the major source of air pollution in urban areas in the past two decades. This article proposes an artificial neural network model for identifying the taxi gross emitters based on the remote sensing data. After carrying out the field test in Guangzhou and analyzing various factors from the emission data, the artificial neural network modeling was proved to be an advisable method of identifying the gross emitters. On the basis of the principal component analysis and the selection of algorithm and architecture, the Back Propagation neural network model with 8-17-1 architecture was established as the optimal approach for this purpose. It gave a percentage of hits of 93%. Our previous research result and the result from aggression analysis were compared, and they provided respectively the percentage of hits of 81.63% and 75%. This comparison demonstrates the potentiality and validity of the proposed method in the identification of taxi gross emitters. PMID- 17915706 TI - Evaluation of ambient air quality in Guangzhou, China. AB - On the basis of the reported air quality index (API) and air pollutant monitoring data provided by the Guangzhou Environment Monitoring Stations over the last twenty-five years, the characteristics of air quality, prominent pollutants, and variation of the average annual concentrations of SO2, NO2, total suspended particulate (TSP), fine particulates (PM10), CO and dustfall in Guangzhou City were analyzed. Results showed that TSP was the prominent pollutant in the ambient air environment of Guangzhou City. Of the prominent pollutants, TSP accounted for nearly 62%, SO2 12.3%, and NOx 6.4%, respectively. The average API of Guangzhou over 6 years was higher than that of Beijing, Tianjin, Nanjing, Hangzhou, Suzhou and Shanghai, and lower than that of Shenzhen, Zhuhai and Shantou. Concentrations of air pollutants have shown a downward trend in recent years, but they are generally worse than ambient air quality standards for USA, Hong Kong and EU. SO2 and NOx pollution were still serious, impling that waste gas pollution from all kinds of vehicles had become a significant problem for environmental protection in Guangzhou. The possible causes of worsening air quality were also discussed in this paper. PMID- 17915707 TI - ATR-FTIR and XPS study on the structure of complexes formed upon the adsorption of simple organic acids on aluminum hydroxide. AB - Information on the binding of organic ligands to metal (hydr)oxide surfaces is useful for understanding the adsorption behaviour of natural organic matter on metal (hydr)oxide. In this study, benzoate and salicylate were employed as the model organic ligands and aluminum hydroxide as the metal hydroxide. The attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectra revealed that the ligands benzoate and salicylate do coordinate directly with the surface of hydrous aluminum hydroxide, thereby forming inner-sphere surface complexes. It is concluded that when the initial pH is acidic or neutral, monodentate and bridging complexes are to be formed between benzoate and aluminum hydroxide while bridging complexes predominate when the initial pH is alkalic. Monodentate and bridging complexes can be formed at pH 5 while precipitate and bridging complexes are formed at pH 7 when salicylate anions are adsorbed on aluminum hydroxide. The X-ray photoelectron (XP) spectra demonstrated the variation of C 1s binding energy in the salicyate and phenolic groups before and after adsorption. It implied that the benzoate ligands are adsorbed through the complexation between carboxylate moieties and the aluminum hydroxide surface, while both carboxylate group and phenolic group are involved in the complexation reaction when salicylate is adsorbed onto aluminum hydroxide. The information offered by the XPS confirmed the findings obtained with ATR-FTIR. PMID- 17915708 TI - A novel acidophile community populating waste ore deposits at an acid mine drainage site. AB - Waste ore samples (pH 3.0) were collected at an acid mine drainage (AMD) site in Anhui, China. The present acidophilic microbial community in the waste ore was studied with 16S rRNA gene clone library and denaturing gradient gel electrophoresis (DGGE). Eighteen different clones were identified and affiliated with Actinobacteria, low G + C Gram-positives, Thermomicrobia, Acidobacteria, Proteobacteria, candidate division TM7, and Planctomycetes. Phylogenetic analysis of 16S rRNA gene sequences revealed a diversity of acidophiles in the samples that were mostly novel. It is unexpected that the moderately thermophilic acidophiles were abundant in the acidic ecosystem and may play a great role in the generation of AMD. The result of DGGE was consistent with that of clone library analysis. These findings help in the better understanding of the generation mechanism of AMD and in developing a more efficient method to control AMD. PMID- 17915709 TI - Comparison of four supports for adsorption of reactive dyes by immobilized Aspergillus fumigatus beads. AB - Four materials, sodium carboxymethylcellulose (Na-CMC), sodium alginate (SA), polyvinyl alcohol (PVA), and chitosan (CTS), were prepared as supports for entrapping fungus Aspergillus fumigatus. The adsorption of synthetic dyes, Reactive Brilliant Blue KN-R, and Reactive Brilliant Red K-2BP, by these immobilized gel beads and plain gel beads was evaluated. The adsorption efficiencies of Reactive Brilliant Red K-2BP and Reactive Brilliant Blue KN-R by CTS immobilized beads were 89.1% and 93.5% in 12 h, respectively. The adsorption efficiency by Na-CMC immobilized beads was slightly lower than that of mycelial pellets. But the dye culture mediums were almost completely decolorized in 48 h using the above-mentioned two immobilized beads (exceeding 95%). The adsorption efficiency by SA immobilized beads exceeded 92% in 48 h. PVA-SA immobilized beads showed the lowest adsorption efficiency, which was 79.8% for Reactive Brilliant Red K-2BP and 92.5% for Reactive Brilliant Blue KN-R in 48 h. Comparing the adsorption efficiency by plain gel beads, Na-CMC plain gel beads ranked next to CTS ones. SA and PVA-SA plain gel beads hardly had the ability of adsorbing dyes. Subsequently, the growth of mycelia in Na-CMC and SA immobilized beads were evaluated. The biomass increased continuously in 72 h. The adsorption capacity of Reactive Brilliant Red K-2BP and Reactive Brilliant Blue KN-R by Na-CMC immobilized beads was 78.0 and 86.7 mg/g, respectively. The SEM micrographs show that the surface structure of Na-CMC immobilized bead is loose and finely porous, which facilitates diffusion of the dyes. PMID- 17915710 TI - Investigation of basic properties of fly ash from urban waste incinerators in China. AB - Basic properties of fly ash samples from different urban waste combustion facilities in China were analyzed using as X-ray fluorescence (XRF), scanning electron microscopy (SEM), X-ray diffraction (XRD). The leaching toxicity procedure and some factors influencing heavy metals distribution in fly ash were further investigated. Experimental results indicate that the fly ash structures are complex and its properties are variable. The results of XRF and SEM revealed that the major elements (>10000 mg/kg, listed in decreasing order of abundance) in fly ash are O, Ca, Cl, Si, S, K, Na, Al, Fe and Zn. These elements account for 93% to 97%, and the content of Cl ranges from 6.93% to 29.18 %, while that of SiO2 does from 4.48% to 24.84%. The minor elements (1000 to 10000 mg/kg) include Cr, Cu and Pb. Primary heavy metals in fly ash include Zn, Pb, Cr, Cu etc. According to standard leaching test, heavy metal leaching levels vary from 0 to 163.10 mg/L (Pb) and from 0.049 to 164.90 mg/L (Zn), mostly exceeding the Chinese Identification Standard for hazardous wastes. Morphology of fly ash is irregular, with both amorphous structures and polycrystalline aggregates. Further research showed that heavy metals were volatilized at a high furnace temperature, condensed when cooling down during the post-furnace system and captured at air pollution control systems. Generally, heavy metals are mainly present in the forms of aerosol particulates or tiny particulates enriched on surfaces of fly ash particles. The properties of fly ash are greatly influenced by the treatment capacities of incinerators or the variation of waste retention time in chamber. Fly ash from combustors of larger capacities generally has higher contents of volatile component and higher leaching toxicity, while those of smaller capacities often produce fly ash containing higher levels of nonvolatile components and has lower toxicity. The content of heavy metals and leaching toxicity maybe have no convincing correlation, and high alkali content of CaO greatly contribute to leaching toxicity of heavy metal and acid neutralization capacity against acid rain. PMID- 17915711 TI - Pollutant loads of surface runoff in Wuhan City Zoo, an urban tourist area. AB - The pollutant loads of surface runoff in an urban tourist area have been investigated for two years in the Wuhan City Zoo, China. Eight sampling sites, including two woodlands, three animal yards, two roofs and one road, were selected for sampling and study. The results indicate that pollutants ranked in a predictable order of decreasing load (e.g. animal yard > roof > woodland > road), with animal yards acting as the key pollution source in the zoo. Pollutants were transported mainly by particulate form in runoff. Particulate nitrogen and particulate phosphorous accounted on average for 61%, 78% of total pollutant, respectively, over 13 monitored rainfall events. These results indicate the treatment practices should be implemented to improve particulate nutrient removal. Analysis of the M(V) curve indicate that no first flush effect existed in the surface runoff from pervious areas (e.g. woodland, animal ground yard), whereas a first flush effect was evident in runoff from impervious surfaces (e.g. animal cement yard, roof, road). PMID- 17915712 TI - Sustainability: a view from the wind-eroded field. AB - This article explores the assessment of sustainability in fields subject to wind erosion. In the first part, simple sustainability audits are examined, as of soil depth and nutrients. Direct measurement of these characteristics has many problems, largely because of huge variability in space and time at all scales. Modelling still has its problems, but it may be possible to overcome many of them soon. It is true that wind erosion preferentially removes soil nutrients, but there are imponderables even here. The nutrient balance in many of these soils includes considerable input from dust. In West Africa, it has been shown that the amounts of calcium and potassium that are added in dust are sufficient to fertilize dispersed crops. In mildly acidic sandy soils, such as those found on the widespread palaeoaeolian deposits, much of the phosphorus is fixed and unavailable to plants by the time it is removed by wind erosion, so that erosion has no added downside. Most of the nutrients carried by dust have been shown to travel close to the ground (even when they are attached to dust-sized particles), and so are trapped in nearby fallow strips, and are thus not lost to the farming system. Second, the sustainability of a whole semi-arid farming system is explored. Wind erosion in semi-arid areas (like China, the Sahel and Northwestern Europe) generally takes place on aeolian deposits of the recent geological past. Most of these soils are deep enough to withstand centuries of wind erosion before they are totally lost to production, and some of these soils have greater fertility at greater depth (so that wind erosion may even improve the soil). Finally some remarks are made about environmental change in relation to sustainability. PMID- 17915713 TI - Identification of anthropogenic influences on water quality of rivers in Taihu watershed. AB - Surface water bodies are progressively subjected to stress as a result of anthropogenic activities. This study assessed and examined the impact of human activities on spatial variation in the water quality of 19 rivers in the Taihu watershed. Concentrations of physicochemical parameters of surface water quality were determined at the mouth of each river during the period of 2000-2004. Multivariate statistical techniques were applied to identify characteristics of the water quality in the studied rivers. The results showed that rivers strongly influenced by household wastewater have the highest concentrations of nutrients (TN and TP). Moreover, rivers in the vicinity of a metropolis presented low dissolved oxygen (DO) levels. However, organic-chemical pollution (petroleum and volatile phenolics) was identified with high localization. Two rivers influenced by sewage from industry and ships were distinguished from other rivers with high values of petroleum. The Taige channel, a river located in Changzhou City that is strongly influenced by wastewater from industry, was characterized with an extraordinarily high value of volatile phenolics. Rivers passing through countries, especially through hilly countries were characterized with high DO contents and low nutrient and organic-chemical pollution, suggesting that agriculture puts less pressure on water quality in adjacent rivers. Therefore, more effort should be made in controlling point pollution to restore water quality in rivers adjacent to cities. PMID- 17915714 TI - Effects of climate change on water resources in Tarim River Basin, Northwest China. AB - Based on hydrology, temperature, and precipitation data from the past 50 years, the effects of climate change on water resources in Tarim River Basin in Northwest China were investigated. The long-term trends of the hydrological time series were detected using both parametric and nonparametric techniques. The results showed that the increasing tendency of the temperature has a 5% level of significance, and the temperature increased by nearly 1 degree C over the past 50 years. The precipitation showed a significant increase in the 1980s and 1990s, and the average annual precipitation exhibited an increasing trend with a magnitude of 6.8 mm per decade. A step change occurred in both the temperature and precipitation time series around 1986. The streamflow from the headwater of the Tarim River exhibited a significant increase during the last 20 years. The increase in temperature, precipitation, and streamflow may be attributed to global climate change. PMID- 17915715 TI - Multi-indicator assessment of water environment in government environmental auditing. AB - This study focused on a multi-indicator assessment methodology for governmental environmental auditing of water protection programs. The environmental status of Wuli Lake in China was assessed using the global indicators (driver-status response) developed by the Commission on Sustainable Development, and four additional indicators proposed by the author: water quality, pollution load, aquatic ecosystem status, and lake sediment deposition. Various hydrological, chemical, biological and environmental parameters were used to estimate the values of the indicators for assessment of environmental status of the lake based on time series data sets for twenty years. The indicators proposed can be customized to meeting the needs for particular assessment of water protection programs. This method can be used to evaluate the performance of national environmental protection programs and provide technical support for environmental auditors. PMID- 17915716 TI - Elevated CO2 changes the moderate shade tolerance of yellow birch seedlings. AB - To demonstrate the existence of light thresholds in plant growth and to examine the effects of elevated CO2 on the shade tolerance of a tree species, an experiment consisting of a completely randomized design for a total of 96 yellow birch (Betula alleghaniensis Britton) seedlings was conducted with 3 light levels (2.9%, 7.7%, 26.1% of full sunlight) x 2 CO2 levels (350 and 700 +/- 10 ppm) with 4 replications in a phytotron. The study proved that thresholds exist and they vary in different plant organs. In ambient CO2, the thresholds were 13.3%, 18.7%, 15.0%, 15.2%, and 15.6% of full sunlight for stem, leaf, root, total plant biomass, and the averaged value, respectively. In 700 ppm CO2, the corresponding thresholds were 16.7%, 21.3%, 18.1%, 21.7% and 19.5% for stem, leaf, root, total plant biomass, and the averaged value, respectively. The lowest threshold in the stem is an indicator of the minimal light intensity for regular growth for seedlings of this species. Below this threshold, light-stressful growth occurs. The result of a paired t-test indicated that the thresholds in elevated CO2 were significantly higher than in ambient CO2. This suggests that yellow birch will lose its moderate shade tolerance, evolutionally becoming a shade-intolerant species, and that it may become more difficult to naturally regenerate in the future. PMID- 17915717 TI - Evaluation of oasis ecosystem risk by reliability theory in an arid area: a case study in the Shiyang River Basin, China. AB - Ecosystem risk is a new concept in understanding environmental problems. It is important to study and develop quantitative methods for regional ecosystem risk analysis. In this study, some new indicators and methods for measuring oasis ecosystem risk were established using reliability theory. These indicators are linked to water resource, which is the key restricting factor in arid area oasis ecosystems. They have clear meanings and can also be compared in different arid area oases. A case study in the Liangzhou oasis of the Shiyang River Basin in China shows how to calculate these ecosystem risk indicators. The results of the case study are as follows: the reliability indicator, risk indicator, stability indicator, and integrated loss indicator of the Liangzhou oasis are 0.686, 0.314, 0.743, and 0.301, respectively. This means that the reliability degree of the oasis's ecosystem safety is 68.6%; the degree of risk that it is unsafe is 31.4%; the stability degree is 74.3%; and 30.1% of the oasis's area is supported by over exploiting underground water and damaging the lower reaches of the ecosystem. This result can be used as a guide in controlling and managing ecosystem risk in the research area. PMID- 17915718 TI - Feasibility and simulation model of a pilot scale membrane bioreactor for wastewater treatment and reuse from Chinese traditional medicine. AB - The lack and pollution of water resource make wastewater reuse necessary. The pilot scale long-term tests for submerged membrane bioreactor were conducted to treat the effluents of anaerobic or aerobic treatment process for the high strength Chinese traditional medicine wastewater. This article was focused on the feasibility of the wastewater treatment and reuse at shorter hydraulic retention time (HRT) of 5.0, 3.2 and 2.13 h. MLSS growth, membrane flux, vacuum values and chemical cleaning periods were also investigated. The experimental results of treating two-phase anaerobic treatment effluent demonstrated that the CODfilt was less than 100 mg/L when the influent COD was between 500-10000 mg/L at HRT of 5.0 h, which could satisfy the normal discharged standard in China. The experimental results to treat cross flow aerobic reactor effluent demonstrated that the average value of CODfilt was 17.28 mg/L when the average value of influent COD was 192.84 mg/L at HRT of 2.13 h during 106 d, which could completely meet the normal standard for water reuse. The maximum MLSS and MLVSS reached 24000 and 14500 mg/L at HRT of 3.2 h respectively. Membrane flux had maximal resume degrees of 94.7% at vacuum value of 0.02 MPa after cleaning. Chemical cleaning periods of membrane module were 150 d. A simulation model of operational parameters was also established based on the theory of back propagation neural network and linear regression of traditional mathematical model. The simulation model showed that the optimum operational parameters were suggested as follows: HRT was 5.0 h, SRT was 100 d, the range of COD loading rate was between 10.664-20.451 kg/(m3xd), the range of MLSS was between 7543-13694 mg/L. PMID- 17915719 TI - Adsorption of phenolic compounds from aqueous solutions by aminated hypercrosslinked polymers. AB - Two novel polymers (NJ-1 and NJ-2) were synthesized by chemically modified a hypercrosslinked polymer NJ-0 with dimethylamine and trimethylamine, respectively. The comparison of the adsorption properties of the three polymers toward phenol, resorcin and phloroglucin was made. The study focused on the static equilibrium adsorption behaviors and the adsorption thermodynamics. Freundlich equation was found to fit the adsorption results well. The effect of amino groups introduced onto the surface of the resin and the structure of phenolic compounds on the adsorption were also studied. The hydrogen-bonding interaction and electrostatic interaction could happen between the amino groups and the adsorbates. The adsorption impetus increased as quantity of hydroxyl groups increased, but the adsorption capacity decreased due to the drop of the matching degree of the aperture of resins and the diameter of adsorbate molecules. PMID- 17915720 TI - Sludge concentration dynamic distribution and its impact on the performance of UNITANK. AB - UNITANK is a biological wastewater treatment process that combines the advantages of traditional activated sludge process and sequencing batch reactor, which is divided into Tank A, B and C. In this study, the sludge distribution and its impact on performance of UNITANK were carried out in Liede Wastewater Plant (WWTP) of Guangzhou, China. Results showed that there was a strong affiliation between Tank A and B of the system in sludge concentration distribution. The initial sludge concentration in Tank A could present the sludge distribution of the whole system. The sludge distribution was mainly influenced by hydraulic condition. Unsteady sludge distribution had an impact on variations of substrates in reactors, especially in decisive reactor, and this could lead to failure of system. Settler could partially remove substrates such as COD and NO3-N, but there was adventure of sludge deterioration. The rational initial sludge concentration in Tank A should be 4000-6000 mg/L MLSS. PMID- 17915721 TI - Characterization of surface runoff from a subtropics urban catchment. AB - Characteristics of surface runoff from a 0.14-km2 urban catchment with separated sewer in Macau was investigated. Water quality measurements of surface runoff were carried out on five rainfall events during the period of August to November, 2005. Water quality parameters such as pH, turbidity, TSS, COD, TN, Zn, Pb, and Cu were analyzed. The results show that TN and COD are the major pollutants from surface runoff with mean concentration of 8.5 and 201.4 mg/L, both over 4-fold higher compared to the Class V surface water quality standard developed by China SEPA. Event mean concentration (EMC) for major pollutants showed considerable variations between rainfall events. The largest rainfall event with the longest length of antecedent dry weather period (ADWP) produced the highest EMC of TN, TSS and COD. From the pollutographs analysis, the peak concentration of TN precedes the peak runoff flow rate for all three rainfall events. The tendency of the concentration of TSS, turbidity and COD changing with runoff flow varies between rainfall events. The relationship between TSS and other parameters were analyzed to evaluate the efficiency of the physical treatment process to control the surface runoff in the urban catchment. Based on the correlation of parameters with TSS, high treatment efficiency of TSS, TN and COD was expected. The most significant event in term of first flush is the one with the strongest rainfall intensity and longest length of ADWP. TN always showed first flush phenomenon in all three rainfall events, which suggested that the surface runoff in the early stage of surface runoff should be dealt with for controlling TN losses during rainfall events. PMID- 17915722 TI - Treatment of coke plant wastewater by SND fixed biofilm hybrid system. AB - In this article, coke plant wastewater was treated by a simultaneous nitrifying and denitrifying (SND) fixed biofilm hybrid system. The results showed that suitable parameters of the system were important for the performance of the bio degradation system. The chemical oxygen demand (COD) removal efficiency in this system was satisfactory, higher than 94%, and ammonia nitrogen was higher than 95%. The effluent COD concentration could meet the discharge standard, except for very few situations. The results showed that a sufficient carbon source was important for making ammonia nitrogen concentration meet the discharge standard. Then the TN removal efficiency in this system can be brought higher than 94%. Dissolved oxygen (DO) is very important to the performance of the SND bio degradation system, and the suitable DO is about 3.5-4.0 mg/L at the forepart of reactor. In addition, the performance of the system was almost not affected by pH value. The results show that the system is feasible to treat coke plant wastewater. PMID- 17915723 TI - Removal of heavy metals from aqueous solution by sawdust adsorption. AB - The adsorption of lead, cadmium and nicel from aqueous solution by sawdust of walnut was investigated. The effect of contact time, initial metal ion concentration and temperature on metal ions removal has been studied. The equilibrium time was found to be of the order of 60 min. Kinetics fit pseudo first-order, second-order and intraparticle diffusion models, hence adsorption rate constants were calculated. The adsorption data of metal ions at temperatures of 25, 45 and 60 degrees C have been described by the Freundlich and Langmuir isotherm models. The thermodynamic parameters such as energy, entropy and enthalpy changes for the adsorption of heavy metal ions have also been computed and discussed. Ion exchange is probably one of the major adsorption mechanisms for binding divalent metal ions to the walnut sawdust. The selectivity order of the adsorbent is Pb(II) approximately Cd(II)>Ni(II). From these results, it can be concluded that the sawdust of walnut could be a good adsorbent for the metal ions from aqueous solutions. PMID- 17915724 TI - Source profiles of particulate organic matters emitted from cereal straw burnings. AB - Cereal straw is one of the most abundant biomass burned in China but its contribution to fine particulates is not adequately understood. In this study, three main kinds of cereal straws were collected from five grain producing areas in China. Fine particulate matters (PM2.5) from the cereal straws subjected to control burnings, both under smoldering and flaming status, were sampled by using a custom made dilution chamber and sampling system in the laboratory. Element carbon (EC) and organic carbon (OC) was analyzed. 141 compounds of organic matters were measured by gas chromatography-mass spectrum (GC-MS). Source profiles of particulate organic matters emitted from cereal straw burnings were obtained. The results indicated that organic matters contribute a large fraction in fine particulate matters. Levoglucosan had the highest contributions with averagely 4.5% in mass of fine particulates and can be considered as the tracer of biomass burnings. Methyloxylated phenols from lignin degradation also had high concentrations in PM2.5, and contained approximately equal amounts of guaiacyl and syringyl compounds. beta-Sitostrol also made up relatively a large fraction of PM2.5 compared with the other sterols (0.18%-0.63% of the total fine particle mass). Normal alkanes, PAHs, fatty acids, as well as normal alkanols had relatively lower concentrations compared with the compounds mentioned above. Carbon preference index (CPI) of normal alkanes and alkanoic acids showed characteristics of biogenic fuel burnings. Burning status significantly influenced the formations of EC and PAHs. The differences between the emission profiles of straw and wood combustions were displayed by the fingerprint compounds, which may be used to identify the contributions between wood and straw burnings in source apportionment researches. PMID- 17915725 TI - Gaseous elemental mercury concentration in atmosphere at urban and remote sites in China. AB - An investigation of gaseous elemental mercury concentration in atmosphere was conducted at Beijing and Guangzhou urban, Yangtze Delta regional sites and China Global Atmosphere Watch Baseline Observatory (CGAWBO) in Mt. Waliguan of remote continental area of China. High temporal resolved data were obtained using automated mercury analyzer RA-915+. Results showed that the overall hourly mean Hg0 concentrations in Mt. Waliguan were 1.7+/-1.1 ng/m3 in summer and 0.6+/-0.08 ng/m3 in winter. The concentration in Yangtze Delta regional site was 5.4+/-4.1 ng/m3, which was much higher than those in Waliguan continental background area and also higher than that found in North America and Europe rural areas. In Beijing urban area the overall hourly mean Hg0 concentrations were 8.3+/-3.6 ng/m3 in winter, 6.5+/-5.2 ng/m3 in spring, 4.9+/-3.3 ng/m3 in summer, and 6.7+/ 3.5 ng/m3 in autumn, respectively, and the concentration was 13.5+/-7.1 ng/m3 in Guangzhou site. The mean concentration reached the lowest value at 14:00 and the highest at 02:00 or 20:00 in all monitoring campaigns in Beijing and Guangzhou urban areas, which contrasted with the results measured in Yangtze Delta regional site and Mt. Waliguan. The features of concentration and diurnal variation of Hg0 in Beijing and Guangzhou implied the importance of local anthropogenic sources in contributing to the high Hg0 concentration in urban areas of China. Contrary seasonal variation patterns of Hg0 concentration were found between urban and remote sites. In Beijing the highest Hg0 concentration was in winter and the lowest in summer, while in Mt. Waliguan the Hg0 concentration in summer was higher than that in winter. These indicated that different processes and factors controlled Hg0 concentration in urban, regional and remote areas. PMID- 17915726 TI - Transformation of mercury speciation through the SCR system in power plants. AB - Coal-fired utility boilers are now identified as the largest source of mercury in the United States. There is speculation that the installation of selective catalytic reduction (SCR) system for reduction of NOx can also prompt the oxidation and removal of mercury. In this paper, tests at six full-scale power plants with similar type of the SCR systems are conducted to investigate the effect of the SCR on the transformation of mercury speciation. The results show that the SCR system can achieve more than 70%-80% oxidation of elemental mercury and enhance the mercury removal ability in these units. The oxidation of elemental mercury in the SCR system strongly depends on the coal properties and the operation conditions of the SCR systems. The content of chloride in the coal is the key factor for the oxidization process and the maximum oxidation of elemental mercury is found when chloride content changes from 400 to 600 ppm. The sulfur content is no significant impact on oxidation of elemental mercury. PMID- 17915727 TI - Effects of long-term amendment of organic manure and nitrogen fertilizer on nitrous oxide emission in a sandy loam soil. AB - To understand the effects of long-term amendment of organic manure and N fertilizer on N2O emission in the North China Plain, a laboratory incubation at different temperatures and soil moistures were carried out using soils treated with organic manure (OM), half organic manure plus half fertilizer N (HOM), fertilizer NPK (NPK), fertilizer NP (NP), fertilizer NK (NK), fertilizer PK (NK) and control (CK) since 1989. Cumulative N2O emission in OM soil during the 17 d incubation period was slightly higher than in NPK soil under optimum nitrification conditions (25 degrees C and 60% water-filled pore space, WFPS), but more than twice under the optimum denitrification conditions (35 degrees C and 90% WFPS). N2O produced by denitrification was 2.1-2.3 times greater than that by nitrification in OM and HOM soils, but only 1.5 times greater in NPK and NP soils. These results implied that the long-term amendment of organic manure could significantly increase the N2O emission via denitrification in OM soil as compared to NPK soil. This is quite different from field measurement between OM soil and NPK soil. Substantial inhibition of the formation of anaerobic environment for denitrification in field might result in no marked difference in N2O emission between OM and NPK soils. This is due in part to more rapid oxygen diffusion in coarse textured soils than consumption by aerobic microbes until WFPS was 75% and to low easily decomposed organic C of organic manure. This finding suggested that addition of organic manure in the tested sandy loam might be a good management option since it seldom caused a burst of N2O emission but sequestered atmospheric C and maintained efficiently applied N in soil. PMID- 17915728 TI - Spatial variability of organochlorine pesticides (DDTs and HCHs) in surface soils from the alluvial region of Beijing, China. AB - The spatial variability in the concentrations of 1,2,3,4,5,6 hexachlorocyclohexane (HCH) and 1,1,1-trichloro-2,2-bis-(p-chlorophenyl) ethane (DDT) in surface soils was studied on the basis of the analysis of 131 soil samples collected from the surface layer (0-20 cm depth) of the alluvial region of Beijing, China. The concentrations of total HCHs (including alpha-, beta-, gamma-, and delta-isomers) and total DDTs (including p,p'-DDT, p,p'-DDD, p,p' DDE, and o,p'-DDT) in the surface soils tested were in the range from nondetectable to 31.72 microg/kg dry soil, with a mean value of 0.91, and from nondetectable to 5910.83 microg/kg dry soil, with a mean value of 32.13, respectively. It was observed that concentrations of HCHs in all soil samples and concentrations of DDTs in 112 soil samples were much lower than the first grade (50 microg/kg) permitted in "Environment quality standard for soils in China (GB15618-1995)". This suggests that the pollution due to organochlorine pesticides was generally not significant in the farmland soils in the Beijing alluvial region. In this study, the spatial distribution and trend of HCHs and DDTs were analyzed using Geostatistical Analyst and GS+(513). Spatial distribution indicated how these pesticides had been applied in the past. Trend analysis showed that the concentrations of HCHs, DDTs, and their related metabolites followed an obvious distribution trend in the surface soils from the alluvial region of Beijing. PMID- 17915730 TI - Residual dynamics of thiacloprid in medical herbs marjoram, thyme, and camomile in soil. AB - Thiacloprid is a new insecticide of the chloronicotinyl family. To assess its risk after application, residual characteristics of thiacloprid in marjoram, thyme, and camomile and in soil were studied under field conditions. The active ingredient was extracted from the plant material using a mixture of acetone water. After filtration, the extract was concentrated to the aqueous phase, diluted with water, and portioned against ethyl acetate on a matrix solid phase dispersion column. Thiacloprid was extracted from soil using a mixture of methanol-water, filtered, and reextracted (clean up) with dichloromethane. The residues were quantified using HPLC-MS-MS. The methods were validated by recovery experiments. Thiacloprid residues in marjoram, thyme, and camomile and in soil persisted beyond 10, 14, 14, and 21 d but no residues were detected after 14, 21, 21, and 28 d, respectively. The data obtained in this study indicated that the biexponential model is more suitable than the first-order function to describe the decline of thiacloprid in fresh marjoram, fresh thyme, and dried camomile flowers with half-life (t1/2) of 1.1, 0.7, and 1.2 d, respectively. However, both the first-order function and biexponential model were found to be applicable for dissipation of thiacloprid in soil with almost the same t1/2 values of 3.5 and 3.6 d. The results indicated that thiacloprid dissipates rapidly and does not accumulate in the tested herbs and in soil. PMID- 17915729 TI - Distribution and ecological effect of mercury in Laogang landfill, Shanghai, China. AB - Laogang landfill near Shanghai is the largest landfill in China, and receives about 10000 t of daily garbage per day. Samples of topsoil and plants were analyzed to evaluate mercury pollution from the landfill. For topsoil samples, there were significant correlations among total mercury (HgT), combinative mercury (HgC) and gaseous mercury (HgG), and content of total organic carbon (TOC), but, no significantly relationship was found between Hg content and filling time. Hg content changes in vertical profiles with time showed that the average HgT of profiles 1992, 1996, and 2000 was similar, but their average HgG was quite different. HgT was significantly correlated with HgC in profile 1992 and 2000, and HgT was significantly correlated with HgG in profile 1996. HgG/HgT ratio in profile samples decreased in the order of (HgG/HgT)1992 > (HgG/Hg)1996 >> (Hg/HgT)2000. A simple outline of Hg release in landfill could be drawn: with increasing of filling time, degradation undergoes different biodegradation, accordingly, gaseous mercury goes through small, more, and small proportion to total mercury. Distribution of Hg in plants was inhomogeneous, following the order of leaf > root > stem. The highest value of leaf may be associated with higher atmospheric Hg from landfill. Ligneous plants (e.g. Phyllostachys glanca, Prunus salicina and Ligustrum lucidum) are capable of enriching more Hg than herbaceous plants. PMID- 17915731 TI - Single and joint effects of pesticides and mercury on soil urease. AB - The influence of two pesticides including chlorimuron-ethyl and furadan and mercury (Hg) on urease activity in 4 soils (meadow burozem and phaeozem) was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase (up to 13%-21%) in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship (P<0.05) between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose (ED50) values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations. PMID- 17915732 TI - Effects of chitosan on growth of an aquatic plant (Hydrilla verticillata) in polluted waters with different chemical oxygen demands. AB - Effects of chitosan on a submersed plant, Hydrilla verticillata, were investigated. Results indicated that H. verticillata could prevent ultrastructure phytotoxicities and oxidativereaction from polluted water with high chemical oxygen demand (COD). Superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents in H. verticillata treated with 0.1% chitosan in wastewater increased with high COD (980 mg/L) and decreased with low COD (63 mg/L), respectively. Ultrastructural analysis showed that the stroma and grana of chloroplast basically remained normal. However, plant cells from the control experiment (untreated with chitosan) were vacuolated and the cell interval increased. The relict of protoplast moved to the center, with cells tending to disjoint. Our findings indicate that wastewater with high COD concentration can cause a substantial damage to submersed plant, nevertheless, chitosan probably could alleviate the membrane lipid peroxidization and ultrastructure phytotoxicities, and protect plant cells from stress of high COD concentration polluted water. PMID- 17915733 TI - Biodegradation of phenol by free and immobilized Acinetobacter sp. strain PD12. AB - A new phenol-degrading bacterium with high biodegradation activity and high tolerance of phenol, strain PD12, was isolated from the activated sludge of Tianjin Jizhuangzi Wastewater Treatment Facility in China. This strain was capable of removing 500 mg phenol/L in liquid minimal medium by 99.6% within 9 h and metabolizing phenol at concentrations up to 1100 mg/L. DNA sequencing and homologous analysis of 16S rRNA gene identified PD12 to be an Acinetobacter sp. Polyvinyl alcohol (PVA) was used as a gel matrix to immobilize Acinetobacter sp. strain PD12 by repeated freezing and thawing. The factors affecting phenol degradation of immobilized cells were investigated, and the results showed that the immobilized cells could tolerate a high phenol level and protected the bacteria against changes in temperature and pH. Storage stability and reusability tests revealed that the phenol degradation functions of immobilized cells were stable after reuse for 50 times or storing at 4 degrees C for 50 d. These results indicate that immobilized Acinetobacter sp. strain PD12 possesses a good application potential in the treatment of phenol-containing wastewater. PMID- 17915734 TI - Oxidation inhibition of sulfite in dual alkali flue gas desulfurization system. AB - A laboratory-scale well-mixed thermostatic reactor with continuously blasting air was used to investigate the oxidation inhibition of sulfite in dual alkali flue gas desulfurization (FGD) system. The effects of operating parameters such as pH value and catalyst concentration on the oxidation were studied. Sodium thiosulfate was used in the system, and was found that it significantly inhabited the sulfite oxidation. In the absence of catalyst, sodium thiosulfate at 12.67 mmol/L had an inhibition efficiency of approximately 98%. While in the presence of catalyst, sodium thiosulfate at 26.72 mmol/L had an inhibition efficiency less than 85.0%. The oxidation reaction order of sulfite in the sodium thiosulfate was determined to be -1.90 and -0.55 in the absence and presence of the catalyst, respectively. Apparent activation energy of oxidation inhibition was calculated to be 53.9 kJ/mol. Pilot tests showed that the consumption rate of thiosulfate agreed well with the laboratory-scale experimental results. PMID- 17915735 TI - Immunotoxicity of bisphenol A to Carassius auratus lymphocytes and macrophages following in vitro exposure. AB - Bisphenol A (BPA) is the monomer component of polycarbonate plastics and classified as an endocrine disrupting chemical (EDC). The reproductive toxicity of BPA has been extensively studied in mammals; however, relatively little information is available on the immunotoxic responses of fish to BPA. In this study, we investigated the effects of BPA on the immune functions of lymphocytes and macrophages in Carassius auratus. The effects of BPA were compared with those of two natural steroid hormones, estradiol and hydrocortisone. Proliferation of the two types of cells in response to PHA was measured using colorimetric MTT assay. Macrophage respiratory burst stimulated by Con A was measured using chemiluminescence assay. Results showed that BPA (0.054-5.4 mg/L), estradiol (0.0002-2.0 mg/L) and hydrocortisone (5-50 mg/L) significantly induced Carassius auratus lymphocyte proliferation while higher doses of hydrocortisone (500-5000 mg/L) appeared to be inhibitory. BPA (0.005-50 mg/L), estradiol (0.005-800 mg/L) and hydrocortisone (0.005-500 mg/L) markedly enhanced macrophage proliferation, whereas higher doses of BPA (500-1000 mg/L) appeared to inhibit cell proliferation. Furthermore, higher dosage of BPA (50 mg/L) and hydrocortisone (50 and 500 mg/L) suppressed the macrophages respiratory burst while estradiol is stimulative all the doses tested (0.05-500 mg/L). In conclusion, BPA could have immunotoxicity to Carassius auratus and functional changes of lymphocyte and macrophage in Carassius auratus may be different between low and high dosages. PMID- 17915736 TI - Degradation of metabolites of benzo[a]pyrene by coupling Penicillium chrysogenum with KMnO4. AB - Several main metabolites of benzo[a]pyrene (BaP) formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone (BP 1,6-quinone), trans-7,8-dihydroxy-7,8 dihydrobenzo[a]pyrene (BP 7,8-diol), 3-hydroxybenzo[a]pyrene (3-OHBP), were identified by high-performance liquid chromatography (HPLC). The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that (1) the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; (2) the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; (3) the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% (w/v), concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites. PMID- 17915737 TI - Influence of lactic acid on the two-phase anaerobic digestion of kitchen wastes. AB - To evaluate the influence of lactic acid on the methanogenesis, anaerobic digestion of kitchen wastes was firstly conducted in a two-phase anaerobic digestion process, and performance of two digesters fed with lactic acid and glucose was subsequently compared. The results showed that the lactic acid was the main fermentation products of hydrolysis-acidification stage in the two-phase anaerobic digestion process for kitchen wastes. The lactic acid concentration constituted approximately 50% of the chemical oxygen demand (COD) concentration in the hydrolysis-acidification liquid. The maximum organic loading rate was lower in the digester fed with lactic acid than that fed with glucose. Volatile fatty acids (VFAs) and COD removal were deteriorated in the methanogenic reactor fed with lactic acid compared to that fed with glucose. The specific methanogenic activity (SMA) declined to 0.343 g COD/(gVSSxd) when the COD loading were designated as 18.8 g/(Lxd) in the digester fed with lactic acid. The propionic acid accumulation occurred due to the high concentration of lactic acid fed. It could be concluded that avoiding the presence of the lactic acid is necessary in the hydrolysis-acidification process for the improvement of the two-phase anaerobic digestion process of kitchen wastes. PMID- 17915738 TI - Spatial pattern of impervious surfaces and their impacts on land surface temperature in Beijing, China. AB - Land surface temperature (LST), which is heavily influenced by urban surface structures, is a significant parameter in urban environmental analysis. This study examined the effect impervious surfaces (IS) spatial patterns have on LST in Beijing, China. A classification and regression tree model (CART) was adopted to estimate IS as a continuous variable using Landsat images from two seasons combined with QuickBird. LST was retrieved from the Landsat Thematic Mapper (TM) image to examine the relationships between IS and LST. The results revealed that CART was capable of consistently predicting LST with acceptable accuracy (correlation coefficient of 0.94 and the average error of 8.59%). Spatial patterns of IS exhibited changing gradients across the various urban-rural transects, with LST values showing a concentric shape that increased as you moved from the outskirts towards the downtown areas. Transect analysis also indicated that the changes in both IS and LST patterns were similar at various resolution levels, which suggests a distinct linear relationship between them. Results of correlation analysis further showed that IS tended to be positively correlated with LST, and that the correlation coefficients increased from 0.807 to 0.925 with increases in IS pixel size. The findings identified in this study provide a theoretical basis for improving urban planning efforts to lessen urban temperatures and thus dampen urban heat island effects. PMID- 17915739 TI - Healthy student nurse: brighter tomorrow. PMID- 17915740 TI - Lady with the lamp. PMID- 17915741 TI - Emotional problems among elderly people in a selected old age home. PMID- 17915742 TI - Acute toxicity levels and ethological responses of Channa striatus to fertilizer industrial wastewater. AB - The present paper deals with the toxicity of fertilizer's industrial wastewater on snake head fish Channa striatus (Bloch) previously named as Ophiocephalus sp., at different concentrations viz., 20, 40, 60, 80 and 100 percent on the behavioural changes and mortality. The exposed fish showed hyperactivity viz., jumping, operculum movement, distance travelling, somersaulting at higher concentration such as 60, 80, and 100 percent within 48 hr. At lower concentrations such as 20 and 40 percent exposed fish showed hyperactivities viz., linear movement and eye movement within 48 hr. The LC50 values of industrial wastewater were also determined viz. 89, 75, 70 and 69 percent at different duration of exposure time i.e. 48, 72, 96 and 120 hr respectively. The obtained data revealed that LC50 value decreased with increase of exposure. PMID- 17915743 TI - Structure of aldobiouronic acid and glucuronic acid from Agathis australis degraded gum polysaccharide. AB - Agathis australis gum on acid hydrolysis with sulphuric acid yielded L-arabinose and D-galactose in 1:4 molar ratio with traces of L-fucose. The components of aldobiouronic acid and glucuronic acid were obtained by graded hydrolysis of degraded gum polysaccharide. The derivatives of aldobiouronic acid was obtained as methyl ester methyl glycoside. PMID- 17915744 TI - Modeling individual tree mortality for crimean pine plantations. AB - Individual tree mortality model was developed for crimean pine (Pinus nigra subsp. pallasiana) plantations in Turkey. Data came from 5 year remeasurements of the permanent sample plots. The data comprises of 115 sample plots with 5029 individual trees. Parameters of the logistic equation were estimated using weighted nonlinear regression analysis. Approximately 80% of the observations were used for model development and 20% for validation. The explicatory variables in the model were ratio of diameter of the subject tree and basal area mean diameter of the sample plot as measure of competition index for individual trees, basal area and site index. All parameter estimates were found highly significant (p < 0.001) in predicting mortality model. Chi-square statistics indicate that the most important variable is d / d(q), the second most important is site index, and the third most important predictor is stand basal area. Examination of graphs of observed vs. predicted mortality rates reveals that the mortality model is well behaved and match the observed mortality rates quite well. Although the phenomenon of mortality is a stochastic, rare and irregular event, the model fit was fairly good. The logistic mortality model passed a validation test on independent data not used in parameter estimation. The key ingredient for obtaining a good mortality model is a data set that is both large and representative of the population under study and the data satisfy both requirements. The mortality model presented in this paper is considered to have an appropriate level of reliability. PMID- 17915745 TI - Biofouling attractants from a brown marine alga Ecklonia cava. AB - In recent years, industrial pollutants and the mountain forest fire ashes released into seawater cause damage to the marine environment, mainly it reduces the algal productivity in the inter tidal region. To get recover from the stress due to pollutants and to increase the growth and development of biofouling algae (benthic organisms), Ecklonia cava extract was investigated for its biofouling attracting efficiency. Bioactive guided fractions of E. cava extract derived from column chromatography were tested against spore attachment of a fouling alga, Ulva pertusa. Fraction B showed increased spore attachment rate with a maximum of 92 +/- 5%. This fraction was further analysed on HPLC, GC-Mass and NMR, deduced as pentadecanoic acid. PMID- 17915746 TI - Effect of phenol on haematological components of Indian major carps Catla catla, Labeo rohita and Cirrhinus mrigala. AB - The effect of phenol on haematological components of Indian major carps, Catla catla, Labeo rohita and Cirrhinus mrigala were observed. After exposure to sublethal concentrations of 5.17 mg l(-1), 6.06 mg l(-1) and 6.99 mg l(-1), the number of red blood cells, haemoglobin content and packed cell volume all decreased but the glucose level increased. The order of decrease in the haematological components of the three fish is in the order of Catla catla > Labeo rohita > Cirrhinus mrigala. PMID- 17915747 TI - Morphology and cultural behavior of Botryococcus protuberans with notes on the genus. AB - The green alga Botryococcus protuberans was isolated from its natural environment and its morphology under different cultural conditions was examined. The alga was characterized by a high starch content and reddish oil drops as the assimilatory products. Photosynthetic pigments, Chl a, Chl b, carotenoids and xanthophylls are present. Modification of environmental conditions in modified Chu-10 medium resulted in optimum growth of the alga. Fatty acid composition revealed palmitic acid being the major component, while lauric acid, myristic acid and stearic acid were found in less quantity. PMID- 17915748 TI - Characterization of soil erosion and its implication to forest management. AB - Forests have traditionally been managed to maximize timber production or economic profit, completely neglecting other forest values. Nowadays, however, forests are being managed for multiple uses. The basic requirement of multiple use forestry is to identify and quantify forest values and to determine management objectives. The priorities of management objectives, however, must be decided. In this study, a model predicting the soil loss for multi objective forest management was developed. The model was based on data from remeasurement of permanent sample plots. The data were gathered from 132 sample plots. Approximately 80% of the observations were used for model development and 20% for validation. The model was designed for even aged and uneven aged forests, as well as for forests with mixed and pure species composition. The explicatory variables in the model were mean diameter and number of trees. All parameter estimates were found highly significant (p < 0.001) in predicting soil loss. The model fit and validation tests were fairly good. The soil loss model presented in this paper was considered to have an appropriate level of reliability. It can be used in the overall multi-objective forest management planning, but, it should be limited to the conditions for which the data were gathered. PMID- 17915750 TI - Prevalence of Staphylococcus aureus and Pseudomonas aeruginosa in indoor air flora of a district hospital, Mandya, Karnataka. AB - Two year (1998 and 1999) extensive survey was carried out in Mandya district hospital, Karnataka for a period of three years to monitor the prevalence of Staphylococcus aureus, Pseudomonas aeruginosa (S. aureus and Ps. aeruginosa) from the indoor air samples of operation theater labor room, children's ward, male and female general wards postoperative wards etc. A rotary air sampler loaded with manital salt and Pseudomonas selection agar media strips were used to collect the samples from various sites. The survivallence results revealed the prevalence of S. aureus and Ps. aeruginosa in almost all sampling sites, irrespective of season indicating their long-term survival and consequent threat to hospitalized patients as well the working employees. PMID- 17915749 TI - Effect of salicylic acid on morphological and biochemical attributes in cowpea. AB - Effects of salicylic acid (SA) on seed germination, seedling growth, flowering and biochemical activities were studied out in four cowpea (Vigna unguiculata) genotypes in control environments. The results revealed that both germination and seedling growth were negatively affected by 0.02%. SA application, however did not affect the size of full expanded buds, time of 50% flowering and date of flower initiation. A maximum increase in peroxidase (EC1.11.1.7) activity was observed in UPC 4200 over other genotypes. No significant change in the content of total soluble and intercellular fluid proteins was observed except in UPC 4200 genotype. SA induced accumulation of total soluble sugars more at flowering stage than at seed setting stage. It is evident from the present study that UPC 4200 genotype was more responsive to salicylic acid both in terms of increased peroxidase activity and less negative effect on morphological attributes, thus suggesting its wider use without negative impact on environment as salicylic acid has been reported in plants. PMID- 17915751 TI - Screening of hepatoprotective effect of a herbal mixture against CCl4 induced hepatotoxicity in Swiss albino mice. AB - The hepatoprotective potential of a herbal mixture was evaluated against CCl4 induced liver injury in Swiss albino mice. Liv 52, a commercially available polyherbal hepatoprotective drug was evaluated for comparison. The potential toxicity of the above herbal hepatoprotective agents was also compared. It was observed that there was a reduction in the enzyme biomarkers (Aspartate and Alanine Transaminase) of liver injury in the herbal mixture treated groups, which was similar to the reduction initiated by Liv 52. An increase in glutathione was observed in the herbal mixture treated groups and it was assumed that the herbal mixture protects the liver by virtue of its antioxidant nature along with high regeneration initiation potential. From the study it is also concluded that the herbal mixture is safer than Liv 52. PMID- 17915752 TI - Impact of soil types and petroleum effluents on the earthworm, Eudrilus eugeniae. AB - Earthworm, Eudrilus eugeniae, exposed to different concentrations of dump-site soil and petroleum effluents exhibited different morbidity and mortality responses. Lake sediments caused varied fluctuations in weight over a 20 day exposure period. Colour changes and mortality up to 15% were observed in earthworms cultured in 100% lake sediment, while weight loss, coiling and sluggish movement were observed in 50% lake water. The effects of 100% dump-site soils were more pronounced as 40% death, swelling, body lesions, stiffening, coiling and low reproduction were recorded. Earthworms were useful as an organism in testing the toxicity of dump-site soils and effluent from a petroleum industry. Dump-site soils and soils polluted with petroleum effluent reduced populations of earthworms and this could subsequently affect other components of the ecosystems associated with earthworm activities. PMID- 17915753 TI - Effects of irrigation water qualities on biomass and sugar contents of sugar beet and sweet sorghum cultivars. AB - An experiment involving four qualities of irrigation water two sugar beet and three sweet sorghum cultivars was conducted in a split plot design with four replications at Rudasht Drainage and Reclamation Experiment Station in 1999. The results showed salinity of water has an adverse effect on sugar beet and sweet sorghum biomass. Sweet sorghum cultivar SSV108 had the lowest biomass under all qualities of irrigation water Sweet sorghum cultivar Rio had the maximum biomass with water qualities of 2, 5, and 8 dS m(-1). Sugar beet cultivar 7233 had the maximum biomass with 11 dS m(-1). The effect of irrigation water quality was not significant for sugar characteristics such as brix, pol and purity. However, responses of cultivars on the above parameters were significant and sugar beet cultivars had higher brix, pol and purity and lower invert sugar and starch than sweet sorghum cultivars. In conclusion, sweet sorghum cultivars are not recommended to be irrigated with saline water of more than 8 dS m(-1) for sugar production. Under such condition, they may be suitable to be grown for forage purposes. PMID- 17915754 TI - Seasonal variations in different physico-chemical parameters of the effluents of Century Pulp and Paper Mill, Lal Kuan, Uttarakhand. AB - The present study was undertaken with the objective to study the characteristics of the effluent of Century Pulp and Paper Mill, Lalkuan (Uttarakhand) in different seasons. The variations in the physicochemical characteristics were observed and monitored up to 12 months at three different sites. Mean values of temperature, pH, chlorides and total phenols of the effluent were found below, whereas colour, BOD5, COD and lignin concentrations were above the minimum national standards (MINAS). PMID- 17915755 TI - The household garbage in the western coast region of Turkey and its relationship with the socio-economic characterstics. AB - The aim of this study is to detect the daily amount of household garbage generated from districts within the limits of municipality of Aydin, the amount and contents of recyclable materials and to evaluate the effect of socioeconomic state. In this cross sectional survey, garbage was collected for one day from all of the districts, weighed both totally and after separation into groups. The districts were assessed according to literacy, drinking water sources and toilet indexes. For each district, three indexes were calculated viz. literacy, water and sewage index. Points between 1-3 were given and by adding points of each index, socioeconomic development (SED) score was calculated for each district. The daily amount of garbage produced was 0.91 +/- 0.74 kg/person and recyclable material was 0.08 +/- 0.13 kg/person (6.4%). A significant corelation was found between SED point and amount of recyclable materials (p < 0.05). There was a positive, strong and significant correlation between workplaces in the districts and recyclable garbage (r = 0.597, p < 0.05). In further analysis, number of workplaces increased the daily amount of garbage by 9.9 times (p < 0.001; 95% confidence Interval: 4.538-15.189), whereas population 0.6 times (p < 0.05; 95% confidence interval: 0.407-0.708). PMID- 17915756 TI - Utilization of both benthic macroinvertebrates and physicochemical parameters for evaluating water quality of the stream Cekerek (Tokat, Turkey). AB - This study examines the applicability of five European biotic indices and the Gammarus:Asellus ratio (G:A), compared with the measurement of physicochemical parameters, in order to determine the water quality at ten sites along the Tokat part of Cekerek stream, in Anatolia, Turkey, during the period February 2002 to January 2003. The biological and chemical results are in good agreement with respect to the water quality. In particular, the G:A ratio was calculated to be high at the first three stations and this result was correlated with the ETBI and the Chandler scores. Consequently, the water quality of Cekerek stream was classified as class I for biological and physicochemical data, except for phosphate, ammonia nitrogen, nitrate and nitrite at the last seven stations. The high concentrations of these chemicals probably result from agricultural runoff and urban sewage. In total, 55 taxa of benthic macroinvertebrates were identified from the Cekerek stream during this study period. PMID- 17915757 TI - Sediment texture and nutrients of Arasalar estuary, Karaikkal, south-east coast of India. AB - Sediment samples were collected for texture, composition and nutrient such as organic carbon, total nitrogen and total phosphorus for a period of two years from three stations in Arasalar estuary. The sediment temperature, pH and nutrients were higher during summer season and lower during monsoon season. At station 1, sediment texture was loamy sand during summer and premonsoon season. At station 2, clay type soil was recorded. Whereas, at station 3, clay was observed in summer and premonsoon and then it shifted to sandy loam type soil during monsoon. Sediment nutrients were lower than that of values of various ecosystem of India. PMID- 17915758 TI - A study on major inorganic ion composition of atmospheric aerosols. AB - Atmospheric aerosol samples were collected from Akola and Buldana region covering around 40 sqkm area during October-November 2002 and were analyzed for ten major inorganic ions namely F-, Cl-, NO3-, SO4(2-), PO4(2-), Na+, K+, Ca2+, Mg2+ and NH4+ using ion chromatographic technique. The average mass of aerosols was found to be 225.81 microg/m3 with standard deviation of 31.29 and average total water soluble load of total cations and anions was found to be 4.32 microg/m3. The concentration of ions in samples showed a general pattern as SO4(2-) > NO3- > Cl- > PO4(2-) > F- for anions and Na+ > Ca2+ > NH4+ > Mg2+ > K+ for cations. The overall composition of the aerosols was taken into account to identify the sources. The trend showed higher concentration of sodium followed by calcium, sulfate, nitrate, phosphate and ammoinum and found to be influenced by terrestrial sources. The presence of SO4(2-) and NO3- in aerosols may be due to re-suspension of soil particles. Ca2+, Mg2+ and Cl- are to be derived from soil materials. The presence of NH4+ may be attributed to the reaction of NH3 vapors with acidic gases may react or condense on an acidic particle surface of anthropogenic origin. The atmospheric aerosol is slightly acidic due to neutralization of basicity by SO2 and NO(x). PMID- 17915759 TI - Impact of industrial effluents on the biochemical composition of fresh water fish Labeo rohita. AB - In acute toxicity (96 hr) experiment the fingerlings of freshwater fish Labeo rohita was exposed to tannery, electroplating and textile mill effluents. The LC0 and LC50 concentrations were 15% and 20% for tannery effluents, 3% and 6% for electroplating effluents and 18% and 22% for textile mill effluents respectively. It was found that, electroplating effluent was more toxic than tannery and textile mill wastes. After acute toxicity experiments for different industrial effluents, various tissues viz. gill, liver, muscle and kidney were obtained separately from control, LC0 and LC50 groups. These tissues were used for biochemical estimations. The glycogen content in all the tissues decreased considerably upon acute toxicity of three industrial effluents except muscle in LC50 group of tannery effluent and kidney in LC50 group of textile mill effluent, when compared to control group. The total protein content decreased in all tissues in three effluents except gills in LC50 group of tannery effluent, kidney in LC50 group of electroplating effluent and kidney in LC0 group of textile mill effluent. In general total lipid content decreased in all tissues after acute exposure when compared to control group. The results obtained in the present study showed that, the industrial effluents from tannery, electroplating and textile mills caused marked depletion in biochemical composition in various tissues of the fish Labeo rohita after acute exposure. PMID- 17915760 TI - Effect of isopod parasite, Cymothoa indica on gobiid fish, Oxyurichthys microlepis from Parangipettai coastal waters (South-east coast of India). AB - The present study reported for the first time on the effect of isopod parasite, Cymothoa indica infestation on Oxyurichthys microlepis an ecologically important gobiid fish from Parangipettai coastal environment (South-east coast of India). The loss of weight in host fishes (male 20.47 and female 32.84%) were observed due to parasitism. The weight of uninfested female fish was found to be higher than that of infested one. The calus like thickening developed on the gill arch and gill filaments of host fish due to the persistent irritation caused by the appendages of the parasite. The reduction of gill surface area was observed due to the attachment of the parasites. The maximum reduction noticed in the first gill arch is mainly due to the heavy pressure exerted by the parasite. Details of gross lesions observed in the branchial chamber, buccal cavity and body surface was enumerated. Heavy infestations of parasitic juveniles have the potential to kill small fingerlings. The swimming capacity of the fish was also found to be affected. PMID- 17915761 TI - Pollution tolerance and distribution pattern of plants in surrounding area of coal-fired industries. AB - Higher concentration of SO2 and particulate matters was reported in surrounding areas of coal-fired industries which influences the distribution pattern of plants. Sensitive plant species are abolished from such areas, however, only pollution tolerant species survive under stress conditions. The present study was designed to investigate the vegetation composition around coal-fired industries i.e. brick industries. To categorise plants as sensitive or resistant air pollution tolerance index (APTI) value was calculated. Out of 99 plants studied, Ricinus communis with APTI 81.10 was found to be the most resistant wild plant showing uniform distribution at all the polluted sites. On the other hand, Lepidium sativum with APTI 5.27 was recorded as the most sensitive plant and found to be present only at the less polluted sites. PMID- 17915762 TI - Association studies for agro-physiological and quality traits of triticale x bread wheat derivatives in relation to drought and cold stress. AB - Correlation coefficient analysis conducted on 22 triticale x bread wheat derivatives along with six checks to select true- breeding derivative(s) for future hybridization programme with tolerance to drought and cold stress conditions as well as better quality traits revealed significant correlation of grain yield with spikelets per spike, biological yield, harvest index, leaf area index. Interestingly, the grain yield and drought susceptibility index showed no association. However, with cold tolerance it showed significant positive correlation indicating the desirability of certain plant traits under cold stress. The grain yield exhibited no association with quality traits which might assist in the predictability of high yielding varieties with high protein, total sugars, reducing sugars and non-reducing sugars. Path coefficient analysis revealed that biological yield had the highest positive direct effect on grain yield followed by harvest index, specific leaf weight, stomatal number, 1000 grain weight, stomatal size, spikelets per spike and days to heading. Therefore, indirect selection for these plant traits in order should be exercised in selecting drought tolerant genotypes. Two genotypes (RL-124-2P2 and RL 111P2) were found to be drought and cold tolerant with high grain yield, spikes per plant, spikelets per spike and leaf area index. PMID- 17915763 TI - Effect of natural preservatives on the growth of histamine producing bacteria. AB - Present study deals with the hampering of the growth of histamine producing bacteria (HPB), by using NaCl and spices which are easily available and cheaper cost wise. For this experiment, four strains of HPB viz. Vibrio parahaemolyticus, Bacillus cereus, Pseudomonas aeruginosa and Proteus mirabilis were tested against 1 to 10% concentrations of NaCl and 1 to 5% concentrations of natural preservatives (turmeric, ginger and garlic) in a basal medium. HPB showed different growth rates at different concentrations of NaCl and natural preservatives. V. parahaemolyticus, B. cereus and Ps. aeruginosa showed no growth at 10% concentration. When the HPB growth was tested with garlic, turmeric and ginger extracts, growth of all the bacteria was inhibited by garlic and turmeric extracts at 5% concentration. In ginger, V. parahaemolyticus, B. cereus and P. mirabilis were totally inhibited at 5% concentration. But Ps. aeruginosa showed very less growth at this concentration. PMID- 17915764 TI - Role of cortisol on condition factor in the female freshwater fish, Notopterus notopterus during four reproductive phases. AB - The condition of the female fish, Notopterus notopterus was studied during four phases of the reproductive cycle in control and after cortisol hormone treatment. The condition of the fish including condition factor (K) and somatic condition factor (Ks) was determined based on weight of the body, length of the fish and gonad weight. In control fish the condition of the fish improve during prespawning phase compared to other phases. The hormone cortisol level estimated by radioimmunoassay (RIA) technique was found to be increased in preparatory and decreased during prespawning phase, may be because of the hormone involvement in metabolic activity, vitellogenesis. In cortisol treatment, the condition of the fish decrease during prespawning phase, compared to other phase may be because of extra expenditure of energy for progressing reproductive activity such as vitellogenesis. PMID- 17915765 TI - Protective role of nordihydroguaiaretic acid (NDGA) against the genotoxic damage induced by ethynodiol diacetate in human lymphocytes in vitro. AB - Antioxidants and plant products are reported to reduce the genotoxic damage of steroids. In our present study we have tested different dosages of nordihydroguaiaretic acid (NDGA) against the genotoxic damage induced by ethynodiol diacetate in the presence of S9 mix. Treatments with nordihydroguaiaretic acid (NDGA) results in the reduction of the genotoxic damage. A significant decrease was observed at all the tested doses of NDGA in sister chromatic exchanges of number of abnormal cells. The results suggest a protective role of NDGA against the genotoxic damage. PMID- 17915766 TI - Radical scavenging activity of Trianthema triquetra in male albino rats intoxicated with CCl4. AB - Radical scavenging activity of ethanolic extract of Trianthema triquetra root was investigated against CCl4 in rats. The rats were treated with T. triquetra (100 mg, 200 mg/kg b.w.) for a period of 7 days. Antihepatotoxic effect was studied by assaying the activities of thiobarbituric acid (TBARS), reduced glutathione (GSH), glutathione peroxidase (GPx),catalase (CAT), super oxide dismutase (SOD) and vitamin C (Vit. C). Lipid peroxidation is evidenced by an increase in the value of TBARS and also a distinct diminution in the level of GSH, Vit. C at 200 mg/kg b.w. The activity of antioxidant enzymes, such as GPx, CAT SOD and Vit. E is significantly recovered towards an almost normal level in animals coadministrated with T. triquetra. The maximum protection against CCl4 induced hepatic injury was afforded by the dose of 200 mg/kg b.w. of Trianthema triquetra. PMID- 17915767 TI - Monitoring of cellular enzymes in the serum of electroplating workers at Coimbatore. AB - Chromium compounds are potent toxic and carcinogenic substances. With respect to toxicity, hepatic and renal toxicity have been reported both in workers and in animals exposed to chromium (VI). Chromium (VI) compounds induces DNA damage in vivo and in cultured cells as well as the cytotoxicity evaluated by the leakage of lactate dehydrogenase. The present study reports the cytotoxicity of chrome platers who are employed from 8 to 25 years in electroplating industries at Coimbatore, Tamilnadu. Blood samples were collected and estimated for glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatine phosphokinase (CPK) and total protein in the serum. The study revealed that there is a significant elevation in the level of LDH, ALP, CPK and transaminases and a decrease in total protein in serum. The results of the study suggests that chromium (VI), a hepatotoxic chemical may perhaps damage the plasma membrane resulting in leakage of enzymes in to the serum of chromeplaters. PMID- 17915768 TI - Potable groundwater quality in some villages of Haryana, India: focus on fluoride. AB - The fluoride concentration in ground water was determined in ten villages of Rohtak district of Haryana state (India). The fluoride concentration in the underground water of these villages varied from 0.034-2.09 mg/l. Various other water quality parameters, viz., pH, electrical conductivity, total dissolved salts, total hardness, total alkalinity sodium, potassium, calcium, magnesium, carbonate, bicarbonate, chloride and sulfate were also measured. A systematic calculation of correlation coefficients among different physicochemical parameters indicated considerable variations among the analyzed samples with respect to their chemical composition. Majority of the samples do not comply with Indian as well as WHO standards for most of the water quality parameters measured. Overall water quality was found unsatisfactory for drinking purposes. Fluoride content was higher than permissible limit in 50% samples. PMID- 17915769 TI - Natural tree collectives of pure oriental spruce [Picea orientalis (L.) Link] on mountain forests in Turkey. AB - Distribution area of oriental spruce [Picea orientalis (L.) Link.] in the world is only in the north-east of Turkey and Caucasian. Because of being the semi monopoly tree with respect to its distribution and representing the upper forest line, it is necessary to analyse, evaluate and model the stand structures of oriental spruce forests in Turkey. In this research, some sampling plots were selected in timberline and treeline in the subalpine forest zone in Turkey. In these sampling plots some information about occurrence and development of the tree collectives was obtained. A total of 12 sampling plots (6 in timberline and 6 of them in treeline) were studied and horizontal and vertical stand profiles were obtained, while number of trees ranges between 2-86 in the tree collectives in treeline and in timberline 3-12. According to this, area per tree in treeline and in timberline is determined as 1.02 m2 and 3.75 m2 on an average respectively. Mean age of trees to reach breast height is 43 years in treeline sampling plots and 22 years in timberline sampling plots. According to the ratio of h (mean height) / d1.30 (diameter at breast height), stand stability values were calculated and it was determined if the stands were stable on the basis of the sampling plots. Stability values of the sampling plots changed between 33 and 75. PMID- 17915770 TI - Leaf biochemistry of Lycopersicon esculentum Mill. at different stages of plant development as affected by mercury treatment. AB - The effect of mercury (Hg) on the biochemical parameters of Lycopersicon esculentum Mill leaf was studied. Application of mercuric chloride in varying concentrations (0.5, 1.0, 1.5 and 2.0 mM HgCl2 kg(-1) sand) caused significant reduction that went up to 89% and 72% chlorophyll a and chlorophyll b contents respectively (at flowering stage), 69% in carotenoid content, 64% in total soluble protein content and 91% in nitrate reductase activity (all at post flowering stage). The amounts of nitrate and proline increased maximally (151% and 143% respectively) at the flowering stage, whereas total soluble sugar enhanced by 57% at the post-flowering stage. Changes observed in most of the parameters, were concentration dependent. Such studies seem to be able to discover suitable bioindicators of heavy metal pollution. PMID- 17915771 TI - Performance of duckweed (Lemna minor L.) on different types of wastewater treatment. AB - Duckweed (Lemna minor L.) has a wide application in Turkey having suitable climatic conditions. In this study, the growth of duckweed was assessed in laboratory scale experiments. They were fed with municipal and industrial wastewater at constant temprature. COD, total nitrogen (TN), total phosphorus (TP) and ortho-phosphate (OP) removal efficiencies of the reactors were monitored by sampling influent and effluent of the system. Removal efficiency in this study reflects optimal results: 73-84% COD removal, 83-87% TN removal, 70-85% TP removal and 83-95% OP removal. The results show that the duckweed-based wastewater treatment is capable of treating the laboratory wastewater. PMID- 17915772 TI - Effect of polystimulin growth regulators and scion clones on graft success and subsequent growth in Atlantic cedar (Cedrus atlantica Manetti). AB - Atlantic cedar (Cedrus atlantica Manetti) was grown, grafting onto the rootstocks of 2 years old Lebanon cedar (Cedrus libani L.). The mixture of polystimulin (PS) growth regulators was used to determine the effects on graft success and subsequent growth during three growing seasons. Scion clones had no effect on grafts success. PS increased the graft success by 20% in comparison to controls. PS-treated grafts burst their buds 18-20 days earlier than control grafts and increased shoot elongation. The PS-treated grafts had 4-5 cm longer shoots than controls at the end of three growing seasons. Thus, this research indicates the significance of PS-application on graft success and subsequent shoot growth on Atlantic cedar. It suggested that use of PS-treated grafts was more profitable than controls. Polystimulins which were used in small doses contributed significantly to the metabolism of Atlantic cedar seedlings after grafting. PMID- 17915773 TI - Nutrient consumption and production of isoflavones in bioreactor cultures of Pueraria lobata (Willd). AB - This paper reports the successful culture of Pueraria lobata (Willd.) suspension cells in a bioreactor. In vitro culture of this Chinese herb has potential as an alternative production method for industrial applications. Calli of P. lobata obtained from leaf explants were cultured in a 5.0 L bioreactor for two weeks. During this period, the pH of the medium declined from 5.8 to 4.5. By the end of the run about 70% of the sugars and reducing sugars and about 50% of nitrate was consumed. Almost 70% of inorganic phosphate and about 80% of the iron was exhausted. The bioreactor results indicated an isoflavone yield of 328.9 microg/ml, with an increase of about 1.77 fold. The yield of puerarin increased about 2.42 fold and reached 73.4 microg/ml in the bioreactor culture. PMID- 17915774 TI - Artificial UV-B induced changes in pigmentation of marine diatom Coscinodiscus gigas. AB - In vitro studies in marine diatom Coscinodiscus gigas revealed that artificial UV B radiation (313 nm) at a dose level of 0.4W m(-2) for a continuous period of 3 hours in a UV treatment chamber caused disbursement of chromatophores from their normal loci and resulted in clumping / aggregation of chromatophores exhibiting a phenomenon called UV-B induced syntrophism. It is also understood that such clumping could cause only insignificant reduction in photosynthetic oxygen release. PMID- 17915775 TI - Fluorides in groundwater and its impact on health. AB - Fluoride is a naturally occurring toxic mineral present in drinking water and causes yellowing of teeth, tooth problems etc. Fluorspar, Cryolite and Fluorapatite are the naturally occurring minerals, from which fluoride finds its path to groundwater through infiltration. In the present study two groundwater samples, Station I and Station II at Hyderabad megacity, the capital of Andhra Pradesh were investigated for one year from January 2001 to December 2001. The average fluoride values were 1.37 mg/l at Station I and 0.91 mg/l at Station II. The permissible limit given by BIS (1983) 0.6-1.2 mg/l and WHO (1984) 1.5 mg/l for fluoride in drinking water. The groundwaters at Station I exceeded the limit while at Station II it was within the limits. The study indicated that fluoride content of 0.5 mg/l is sufficient to cause yellowing of teeth and dental problems. PMID- 17915776 TI - Studies on fluoride removal using adsorption process. AB - Batch adsorption studies were undertaken to assess the suitability of commercially available activated charcoal to remediate fluoride-contaminated water. The effects of some of the major parameters of adsorption, viz. pH, dose of adsorbent, rate of stirring, contact time and initial adsorbate concentration on fluoride removal efficiency were studied and optimized. The optimum sorbent dose was found to be 2.0 g/100 mL, equilibrium was achieved in 120 minutes and enhanced adsorption was obtained at pH 2. Maximum fluoride removal was observed to be 94% at optimum conditions. Freudlich as well as Langmuir isotherms were plotted and kinetic constants were determined. PMID- 17915777 TI - Statistical assessment of underground drinking water contamination and effect of monsoon at Hasanpur, J. P. Nagar (Uttar Pradesh, India). AB - Water Quality Index (WQI) has been calculated for underground drinking water at Hasanpur in J. P. Nagar district, Uttar Pradesh, India at ten different sites in the pre-monsoon season as well as after the onset of monsoon. Water quality parameters were selected as per the WHO guidelines and seventeen water quality physico-chemical parameters were estimated following the standard methods and procedures. Drinking water at almost all the sites was found to be highly contaminated, except a few sites, where it was found moderately contaminated for both the periods during the year 2005. In general, to some extent water quality showed deterioration after the onset of monsoon. It was found that some effective measures are urgently required for water quality management in this region. PMID- 17915778 TI - Ingress of Bhavnagar City (India). AB - A preliminary survey of the coastal city-Bhavnagar was undertaken to assess salinity ingress probed through groundwater quality. Water samples from the wells and bores located in the study area were collected and analyzed. Bhavnagar City is found significantly affected by the seawater intrusion. The ground water showed very high values of SO4(-2), Cl(-1), PO4(-3) and, Na(+1), K(+1) compared to the permissible limits for drinking purposes. The quality of ground water in some of the areas was found highly saline and can not be used even for irrigation purpose. The results also indicated a gradual encroachment of seawater into the native ground water. PMID- 17915779 TI - Sequencing Batch Reactor (SBR) treatment for simultaneous organic carbon and nitrogen removal--a laboratory study. AB - The performance of a laboratory-scale Sequencing Batch Reactor (SBR) has been studied in aerobic-anoxic sequence for simultaneous organic carbon and nitrogen removal. The reactor was operated under three different variations of aerobic anoxic sequence, viz. 4+4, 5+3 and 3+5 hours with input solutions of soluble COD (SCOD) level 1000 +/- 100 mg/L and initial ammonia nitrogen of 40 and 90 mg/L. It has been observed that 85 to 92% of SCOD removal would be possible at the end of 8.0 hour of overall reaction period, irrespective of the length of the aerobic react period. In the case of 4+4 hour operating cycle, reasonable degree of nitrification (88-100%) and denitrification (73-75%), along with 91-94% of organic carbon removal have been achieved, which has been considered to be the optimum performance of the reactor. PMID- 17915780 TI - Assessment of occupational exposure to VOCs at the Gantry gasoline Terminal. AB - The gasoline components dispersed in the workplace environment have direct impact on human health because of their carcinogenic and mutagenic potential. The volatile portion of these compounds generally consists of a mixture of monoaromatics such as benzene, toluene, ethylbenzene, xylenes and substituted benzene compounds. Investigation of levels of these chemicals at the petroleum liquid storage, and distribution facilities is of growing importance for assessing the occupational exposure of workers at these locations. A continuous monitoring was conducted at the Gantry Terminal of a refinery with portable samplers for determining 8-hr TWA at various locations involving petroleum liquid loading and distribution in road tank trucks, rail cars, and drums. After thermal desorption, the concentrations were determined in the laboratory with advanced Varian Model 2200GC/MS system. TVOCs level ranged between 0.10 to 1.97 mg/m3.The 8-hr TWA for benzene, toluene, and xylenes were low and within the limits for occupational exposure of gasoline components. PMID- 17915781 TI - Heavy metals content in water, water hyacinth and sediments of Lalbagh tank, Bangalore (India). AB - The present study was undertaken for assessing the level of heavy metals such as iron, zinc, copper, nickel, chromium, lead and cadmium in water, water hyacinth and sediment samples of Lalbagh tank, Bangalore. Metals were detected using flame atomic absorption spectrophotometry. The results revealed that by and large all metals are present in all the samples, except cadmium in the sediment samples. The average concentrations of iron in water and sediment samples, and lead in water hyacinth were found exceeding the limits of Indian Standards. In general, the concentrations of iron and zinc were found more, followed by lead, chromium, copper, nickel and cadmium cancentration was low. Bioaccumulation factor for water hyacinth was found maximum for chromium. Geoaccumulation index results revealed that there is moderate input of copper and lead from anthropogenic sources to the tank basin. PMID- 17915782 TI - Degradation of glyphosate in soil and its effect on fungal population. AB - Glyphosate application resulted in a decline in soil pH with consequent increase in soil mycoflora suggesting an indirect relationship. Though the composition of mycoflora unchanged, species of aspergilli, fusaria, penicillia and Trichoderma were predominant. HPLC, IR analysis revealed the presence of sarcosine derivative as an intermediary of glyphosate degradation in soil. PMID- 17915783 TI - Environment quality at Maitri station in Antarctica. AB - A comprehensive study of air, water and soil quality was undertaken during the austral summer of 1999-2000 at the Indian Polar Research Station "Maitri" in compliance with the statutory requirements of the article 3 of Protocol on Environmental Protection to the Antarctic Treaty. The main objective of the study was to assess the impacts of various scientific programs and their associated logistic support facilities on the fragile ecosystem of Antarctica. Identification of major sources of pollution and quantification of pollutants in different environmental components were carried out through an extensive environmental monitoring program spread over a period of 5-7 weeks. Preliminary studies reveal that the levels of pollution are not alarming but there is scope for concern looking into the critical aspects of Antarctic environment and the carrying capacity of the environment surrounding Maitri station. PMID- 17915784 TI - Hydrochemistry of groundwater of Thirumanur area, Tamil Nadu (India). AB - Perambalur is the most backward district in the state of Tamil Nadu (India). It has ten unions, of which Thirumanur union has historical importance and is blessed with fertile lands. Thirumanur union has Kollidam river and Pullambadi canal as rich sources of water for irrigation. Thirumanur union has large number of Chozha temples at various places like Thirumalapadi, Periyamanai, Kamarasavalli, Thoothur, Senapathimudikondan, Kandaradhitham, Keelapalur and Melapalur. Thirumanur union being the south end union of Perambalur district, has Trichy district and Tanjore district as its neighbours. Thirumanur union has an area of 64 hectare, of which 48 hectare is under cultivating land. This union has maximum number of thirty-six villages. Though most of the villages use river water as their major source of drinking, they also use groundwater as their other source. Lot of work has been done and published already on the groundwater quality of many villages in other different unions of the Perambalur district. But in the Thirumanur union, there was need to undertake the study to assess the drinking water quality in the region. Hence, ten villages of Thirumanur union were selected, where the people use groundwater for drinking, and the water samples were subjected to systematic analysis. The depth of the bore wells varied from 100-200 feet. The values obtained for different parameters were compared with the standard values given by ISI / ICMR / WHO and the variations were notable for the parameters like nitrate and total hardness for few samples. Therefore, a medical survey was carried out to study the harmful effects on the society due to these two parameters at the villages-Keelapazhur, Palanganatham and Venganur. PMID- 17915786 TI - Noise pollution in residential areas of Jharsuguda Town, Orissa (India) and its impact. AB - In this paper, an attempt has been made to study noise levels in different residential areas of Jharsuguda town in western Orissa (India). Minimum, maximum, L10, L50 and L90 noise levels have been computed. It was found that noise levels in the residential areas exceed the standards set by the Central Pollution Control Board, India. Vehicular traffic, with air horns of loud noise, was found to be the main reason for these high noise levels. Strict measures need to be taken to reduce and control the noise pollution. PMID- 17915785 TI - Scavenging of Ni(II) metal ions by adsorption on PAC and babhul bark. AB - The removal of toxic nickel metal ions by adsorption, using powder activated charcoal (PAC) and non-conventional adsorbent modified Indian powder babhul bark (PBB), was studied at room temperature. The adsorption isotherms were obtained in a batch reactor. It is observed that, the process of uptake followed first-order adsorption rate expression and obeyed Langmuir and Freundlich models of adsorption. Effects of variations in parameters such as pH, contact time, adsorbent dose, initial Ni(II) concentration and particle size were also studied. PMID- 17915788 TI - The use of testa of groundnut shell (Arachis hypogea) for the adsorption of Ni(II) from the aqueous system. AB - This paper presents the use of testa of groundnut shell for the removal of nickel from dilute aqueous solutions at laboratory scale. The adsorption isotherm of Ni(II) on the testa of groundnut shell was carried out by the batch adsorption process. Various parameters such as initial concentration, pH and amount of the adsorbent doses were studied. The experimental adsorption data obtained followed both Langmuir as well as Freundlich isotherms. Maximum adsorption (85%) was observed at pH 6. The monolayer adsorption capacity was found to be 18.79 mg/g, which is greater than most of the low-cost adsorbents reported. PMID- 17915787 TI - Performance evaluation of common effluent treatment plant for tanneries at Pallavaram CETP. AB - The Pallavaram Tanners Industrial Effluent Treatment Company Limited has been a forerunner in ensuring cleaner and safe environment. The project was implemented by a company formed by the tanners of the Pallavaram area. Tamil Nadu Pollution Control Board (TNPCB) was the counterpart of UNIDO and took the overall responsibility for implementation of the project. The present study has been undertaken to evaluate performance efficiency of the treatment plant. Water samples were collected at different stages of treatment units and analysed for the major water quality parameters, such as pH, biological oxygen demand (BOD), chemical oxygen demand (COD), total suspended solids (TSS) and total dissolved solids (TDS). The performance efficiency of each unit in treating the pollutants was calculated. The generated data presented evidence to that the common effluent treatment plant has been working with the norms of TNPCB and meeting the discharge standard limits. PMID- 17915789 TI - Study on leaching of pollutants from vegetable tanning residue. AB - The processing of heavy leather employs the vegetable tanning method involving use of tan liquor. The solid residue of this vegetable tanning process aggravates the water pollution by means of leaching of tannin and other associatedpollutants. Tannin is a biologically resistant compound causing several problems in animal body. The present study dealt with the pollution hazard arising out of leaching of pollutants from the open dumped vegetable tanning residue by rainfall washing. The concerned pollutants were pH, Chemical Oxygen Demand (COD), tannin,sulfate and chloride concentration. To explore the possible extent of contamination of these pollutants in the leached water, two different masses (2 kg and 3 kg) of tanning residue were employed. The results of the study showed that there was a continuous release of pollutants from the vegetable tanning residue. Moreover, there was no regular variation in various pollutant concentrations in both the cases solely due to non-homogeneity of the residue. PMID- 17915790 TI - Third party reproduction in recent scenario. PMID- 17915791 TI - Epilepsy awareness among parents of school children--a municipal survey. AB - A pilot study was undertaken to assess the knowledge, attitude and practice of parents of school-going children in a semi-urban municipal area of Bengal. An open-ended questionnaire with 14 questions were put in local language. A total of 1068 parents were interviewed with 76.9% males and 23.1% females. Most of them (60.7%) having an educational level above primary but below higher secondary school level; 41.4% surveyed believe epilepsy is caused by an infection and 40.8% have the idea that it is contagious too; 80.3% have the idea that epilepsy can be treated but 7.3% interviewed carry the idea that it cannot be treated at all. Only 35.9% responders are ready to allow an epileptic child to attend school and sit together with other children. In terms of first-aid management only 36.6% mentioned some useful first-aid measure. The general public attitude is reflected in this study. False belief and negative attitude leads to barrier in children education. Larger and comprehensive community-based educational programme is very much essential to bring about a change in negative attitude towards epilepsy. PMID- 17915792 TI - Malarial hepatitis as a component of multiorgan failure--a bad prognostic sign. AB - A prospective study among the patients (n = 301) belonging to the coastal districts of Orissa having complicated falciparum malaria with multiorgan failure fulfilling modified APACHE II criteria, for a period of two years in this hospital setting was carried out with particular emphasis on hepatic involvement. There were 206 males and the rest females. Hepatic involvement in the form of raised serum bilirubin levels > or = 6 mg% and prothombin time > 4 compared to controls was found in 192 cases (63.8%). On analysis out of 192 cases predominantly conjugated hyperbilirubinaemia, mixed patterns and unconjugated hyperbilirubinaemia were seen in 115 (59.9%), 64 (33.3%) and 13 (6.8%) cases respectively. Serum bilirubin ranged from 6 to 38 mg%. Aminotransferase aspartate (AST, SGOT) and aminotransferase alkaline (ALT, SGPT) were raised almost two-fold in 98% cases of multiorgan failure with hepatic failure with mean values of 78 +/ 30.4 IU/l and 81 +/- 29.06 IU/l respectively. Nearly three-fold elevation of alkaline phosphatase was observed in 80% cases with mean (315 +/- 39.4 IU/l). Prothombin time was prolonged with mean 7 +/- 3 seconds. Serum proteins and albumin/globulin ratio were normal. There was no difference in glycaemic status over controls. In selected cases, liver histopathological study showed abnormalities in the form of Kupffer cell hyperplasia, mononuclear cell infiltration, hepatocyte necrosis, fatty changes and cholestasis. Majority of patients in multiorgan failure who died, had hepatic failure. PMID- 17915793 TI - Postcoital intra-uterine aspirate--a unique criterion for selecting patients for natural or assisted conception. AB - The success of any infertility management programme is reflected in its efficacy to achieve a pregnancy either spontaneously or through assistance. For this purpose some routine investigations are carried out to pinpoint the nature of problem(s) which an infertile couple is troubled with. As the basic requisite for conception to occur is the accumulation of motile spermatozoa around the egg at the site of fertilisation, proper assessment of sperm transit from the vagina to the tubal ampulla becomes a very important criterion to predict the possibility of spontaneous pregnancy. Till date postcoital test was the sole investigative tool available to serve this purpose but unfortunately it fails to furnish any information about the concentration and nature of quality of spermatozoa beyond the cervix. This results in the disparity between postcoital test results and pregnancy outcome. In the present study intra-uterine fluid was examined along with cervical mucus 4-6 hours postcoitus in the pre-ovulatory period. The objective was to gather information about the fate of spermatozoa subjected to the uterine environment after being vaginally deposited during coitus. The result of this combined test has been found to be very much informative and helpful for the fertility physician to set the criteria for timed intercourse and / or intra uterine insemination. This simple test thus appears to play a leading role in infertility management in near feature. PMID- 17915794 TI - A study on the effects of diclofenac sodium and etoricoxib in the treatment of osteoarthritis. AB - Diclofenac sodium, a non-selective cyclo-oxygenase inhibitor and etoricoxib, a selective cyclo-oxygenase-2 inhibitor have been widely used in treatment of patients with osteo-arthritis. Five hundred and eighty-five patients with uncomplicated knee osteo-arthritis were randomly allocated into 3 equal groups and received either diclofenac sodium, etoricoxib or placebo in a double-blind manner. The response in both the drug groups was comparable and much more than placebo group. The study shows that etoricoxib provides better clinical efficacy and gastro-intestinal tolerability in osteo-arthritis in comparison to diclofenac sodium presumably due to the selective inhibition of cyclo-oxygenase-2 by etoricoxib. PMID- 17915795 TI - Common orthopaedic problems in day to day general practice (in economically handicapped community). AB - The object for writing this article is to summarise different common orthopaedic problems for which common people of our country, economically handicapped, come to the general practitioners in their day to day practice. 'Pain' is the main ailment whatever may be the cause. Where pain killer is not the answer, it is to aware the general practitioner to find out what is behind the pain. A ready reference knowledge for that condition with its available treatment modalities is essential for the doctor and the patient. Deformities and other complaints are mentioned after the pain. An attempt has been made to classify the causes and in short the treatment modalities according to the disease and site to be addressed. Influence of information media on public opinion for decision making has gained momentum. To gain more knowledge, the general practitioners, who are the pillars, the pillars of national health, should study a little more in detail the Journal of Indian Medical Association (JIMA), available textbooks or other journals to have a first hand knowledge to tackle orthopaedic cases. PMID- 17915796 TI - Solitary neurilemmoma in postaural region. AB - Neurilemmoma in postaural region arising from great auricular nerve is an extremely rare tumour. An 11 years boy presented with pain and swelling behind his left ear for last 3-4 years. The clinical examination revealed the swelling appeared to be diffuse with the margin being ill defined. On radiological examination a diffuse homogeneous mass was seen in the postaural region of the left side. The tumour was completely removed by an incision through postaural route. Histopathological study revealed neurilemmoma. Postoperative period was uneventful. PMID- 17915797 TI - Myotonic dystrophy with pregnancy. AB - Myotonic dystrophy is a rare heredodegenerative muscular disorder in which pregnancy is unusual. Because of the autosomal dominant inheritance of the disease, 50% of children of an affected parent may have the disease; 20% of them are asymptomatic at birth. Foetal involvement may be manifested by polyhydramnios, arthrogryposis multiplex in utero, respiratory difficulties, and floppiness at birth. A case of myotonic dystrophy with pregnancy is presented here. PMID- 17915798 TI - EVOLVE (nebivolol evaluation for efficacy and safety in the treatment of hypertension) postmarketing surveillance study. AB - The objective of EVOLVE [nebivolol (nevol) evaluation for efficacy and safety in the treatment of hypertension], a postmarketing surveillance (PMS) study is to identify, validate and quantify the safety and efficacy associated with the use of nebivolol. EVOLVE study was an open-label, non-comparative, prospective, one month follow-up study of 301 patients of either sex with stage 1 hypertension, as defined by the JNC VII guidelines. The data was collected from 27 centres from all over India during the period August, 2006 to December, 2006. Nebivolol (2.5-5 mg/day) was given for 1 month. Clinical assessment was done at the start of the treatment and at 15th day and 30th day follow-ups. Concomitant medications administered were also recorded. Baseline mean systolic blood pressure (SBP) was 157.73 +/- 14.16 mm Hg which dropped to 135.13 +/- 11.15 mm Hg at the end of the study. At the end of 1 month treatment the change in mean SBP was 22.6 mm Hg ie, 14.32% reduction from baseline which was statistically significant (p < 0.001). Also the baseline mean diastolic blood pressure (DBP) was 97.21 +/- 8.25 mm Hg that dropped to 83.69 +/- 6.63 mm Hg at the end of the study. At the end of one month treatment the change in mean DBP was 13.52 mmHg ie, 13.9% reduction from baseline which was significant (p < 0.001). The heart rate in this study showed a significant decrease from 86.13 +/- 9.35 at basal to 75.09 +/- 7.42 at the end of the study (p < 0.001). It was observed that at the end of one month of treatment, majority of the patients ie, 97.75% of total cases showed good to excellent response to nebivolol. EVOLVE PMS study showed that nebivolol hydrochloride is very safe and only 8.2% of cases (n = 22) reported adverse effects, the commonest being dizziness (3.28%). Less than 1% patients reported nausea, constipation, headache, weakness, tiredness and pedal oedema; 99.25% of patients reported good to excellent tolerability; 82.33% patients achieved the goals recommended by JNC VII. EVOLVE PMS study confirms the safety and efficacy of nebivolol hydrochloride in Indian population. PMID- 17915799 TI - A randomised double-blind study comparing sodium feredetate with ferrous fumarate in anaemia in pregnancy. AB - Iron deficiency anaemia is a major health problem in India especially in women of reproductive age group. The World Health Organisation recommends that the haemoglobin concentration should not fall below 11.0 g/dl at any time during pregnancy. The aim of study was to compare the efficacy and safety of two doses of sodium feredetate with ferrous fumarate in improving haemoglobin profile in pregnant anaemic women. Pregnant women with gestation period between 12 and 26 weeks having serum haemoglobin < 10 g/dl, serum ferritin levels less than 12 microg/l were included in the study. Patients were divided into 3 groups and drugs administered accordingly. A total of 48 patients were available for analysis which included 37 patients who had completed all the visits up to 75 days follow-up and 11 patients who were treatment failures. In group A combination of sodium feredetate (containing 33 mg of elemental iron) along with vitamin B12 (15 microg) and folic acid (1.5 mg) was administered twice a day. In group B combination of sodium feredetate (containing 66 mg of elemental iron) along with vitamin B12 (15 microg) and folic acid (1.5 mg) was administered twice a day. In group C combination of ferrous fumarate (containing 100 mg of elemental iron) along with vitamin B12 (15 microg) and folic acid (1.5 mg) was administered twice a day. Patients were evaluated for Hb, RBC count, MCV, MCH and MCHC at day 0, 30, 45, 60 and 75. Serum ferritin, serum iron, TIBC and transferrin saturation were assessed at recruitment and end study. Mean rise of haemoglobin at the completion of study, over that of basal values was 1.79 g/dl (0.71 to 2.87, 95% CI, p < 0.05) in group A, 1.84 g/dl (0.82 to 2.86, 95% CI, p < 0.05) in group B and 1.63 g/dl (0.38 to 2.88, 95% CI, p < 0.05) in group C. Safety assessment was done by doing liver and kidney function test at the time of recruitment and end study. Low doses of sodium feredetate (33 mg and 66 mg of elemental iron given twice daily) produce comparable results as higher dose of ferrous fumarate (100 mg elemental iron given twice daily). As there were no adverse effects reported with sodium feredetate, it can be concluded from this study that this new formulation appears to be effective in improving haemoglobin profile in pregnant anaemic women and is tolerated well. PMID- 17915800 TI - Principles of geriatric pharmacology. AB - Elderly patients are affected by multiple chronic age-related diseases. In prescribing drugs for them one should have sound knowledge of the physiological changes and social issues. Altered drug behaviour in the elderly is to be taken into account. Increasing age may alter absorption, distribution, metabolism or excretion of drugs which have been discussed in a nutshell. While prescribing drugs to the elderly one must be cautious in selection, right doses, proven efficacy, polypharmacy, cost factor and so on. PMID- 17915801 TI - Modification of theophylline release with alginate gel formed in hard capsules. AB - The aim of this work was to establish whether alginate gel formed spontaneously in hard gelatin capsules which modifies release of a model drug, theophylline. The effects of the alginate composition, the calcium addition, and the dissolution medium on drug release were also investigated. After the capsule shell dissolved in water, at neutral pH the gel layer of sodium alginate was formed immediately as the sodium alginate hydrated and swelled on contact with the aqueous medium. In acidic pH, the contents remained intact and the matrix shape was the same. Theophylline release from capsules containing different grades of alginate demonstrated different release patterns, depending on alginate composition and the pH of the medium. The capsules containing sodium/calcium salts of alginate showed the slowest drug release at neutral pH but the fastest in acidic medium. The presence of calcium acetate in the formulations influenced the drug release kinetics. The drug release in acidic medium showed a non-Fickian diffusion-controlled release, while those in water at neutral pH exhibited a Super Case II transport mechanism. The study also provides evidence that the behavior of alginate in forming the hydrated gel layer may explain the drug release behavior at different pHs. PMID- 17915802 TI - Monitoring ibuprofen release from multiparticulates: in situ fiber-optic technique versus the HPLC method: a technical note. PMID- 17915803 TI - Formulation and optimization of porous osmotic pump-based controlled release system of oxybutynin. AB - The aim of the current study was to design a porous osmotic pump-based drug delivery system for controlled release of oxybutynin. The porous osmotic pump contains pore-forming water-soluble additives in the coating membrane, which after coming in contact with water, dissolve, resulting in an in situ formation of a microporous structure. The dosage regimen of oxybutynin is one 5-mg tablet 2 to 3 times a day. The plasma half-life ranges from approximately 2 to 3 hours. Hence, oxybutynin was chosen as a model drug with an aim to develop a controlled release system for a period of 24 hours. Linear and reproducible release similar to that of Ditropan XL was achieved for optimized formulation (f2 >50) independent of hydrodynamic conditions. The effect of different formulation variables, namely, ratio of drug to osmogent, membrane weight gain, and level of pore former on the in vitro release was studied. Cellulose acetate (CA) was used as the semipermeable membrane. It was found that drug release rate increased with the amount of osmogent because of the increased water uptake, and hence increased driving force for drug release. Oxybutynin release was inversely proportional to the membrane weight gain; however, directly related to the level of pore former, sorbitol, in the membrane. This system was found to deliver oxybutynin at a zero order rate for 20 hours. The effect of pH on drug release was also studied. The optimized formulations were subjected to stability studies as per International Conference on Harmonisation (ICH) guidelines and formulations were stable after a 3 month study. PMID- 17915804 TI - Mucoadhesive bilayered tablets for buccal sustained release of flurbiprofen. AB - The aim of this work was the design of sustained-release mucoadhesive bilayered tablets, using mixtures of mucoadhesive polymers and an inorganic matrix (hydrotalcite), for the topical administration of flurbiprofen in the oral cavity. The first layer, responsible for the tablet retention on the mucosa, was prepared by compression of a cellulose derivative and polyacrylic derivative blend. The second layer, responsible for buccal drug delivery, was obtained by compression of a mixture of the same (first layer) mucoadhesive polymers and hydrotalcite containing flurbiprofen. Nonmedicated tablets were evaluated in terms of swelling, mucosal adhesion, and organoleptic characteristics; in vitro and in vivo release studies of flurbiprofen-loaded tablets were performed as well. The best results were obtained from the tablets containing 20 mg of flurbiprofen, which allowed a good anti-inflammatory sustained release in the buccal cavity for 12 hours, ensuring efficacious salivary concentrations, and led to no irritation. This mucoadhesive formulation offers many advantages over buccal lozenges because it allows for reduction in daily administrations and daily drug dosage and is suitable for the treatment of irritation, pain, and discomfort associated with gingivitis, sore throats, laryngopharyngitis, cold, and periodontal surgery. Moreover, it adheres well to the gum and is simple to apply, which means that patient compliance is improved. PMID- 17915805 TI - Transbuccal delivery of 5-aza-2 -deoxycytidine: effects of drug concentration, buffer solution, and bile salts on permeation. AB - Delivery of 5-aza-2 -deoxycytidine (decitabine) across porcine buccal mucosa was evaluated as an alternative to the complex intravenous infusion regimen currently used to administer the drug. A reproducible high-performance liquid chromatography method was developed and optimized for the quantitative determination of this drug. Decitabine showed a concentration-dependent passive diffusion process across porcine buccal mucosa. An increase in the ionic strength of the phosphate buffer from 100 to 400 mM decreased the flux from 3.57 +/- 0.65 to 1.89 +/- 0.61 microg/h/cm2. Trihydroxy bile salts significantly enhanced the flux of decitabine at a 100 mM concentration (P > .05). The steady-state flux of decitabine in the presence of 100 mM of sodium taurocholate and sodium glycocholate was 52.65 +/- 9.48 and 85.22 +/- 7.61 microg/cm2/h, respectively. Two dihydroxy bile salts, sodium deoxytaurocholate and sodium deoxyglycocholate, showed better enhancement effect than did trihydroxy bile salts. A 38-fold enhancement in flux was achieved with 10 mM of sodium deoxyglycocholate. PMID- 17915806 TI - Design and development of hydrogel beads for targeted drug delivery to the colon. AB - The purpose of this research was to develop and evaluate a multiparticulate system of chitosan hydrogel beads exploiting pH-sensitive property and specific biodegradability for colon-targeted delivery of satranidazole. Chitosan hydrogel beads were prepared by the cross-linking method followed by enteric coating with Eudragit S100. All formulations were evaluated for particle size, encapsulation efficiency, swellability, and in vitro drug release. The size of the beads was found to range from 1.04 +/- 0.82 mm to 1.95 +/- 0.05 mm. The amount of the drug released after 24 hours from the formulation was found to be 97.67% +/- 1.25% in the presence of extracellular enzymes as compared with 64.71% +/- 1.91% and 96.52% +/- 1.81% release of drug after 3 and 6 days of enzyme induction, respectively, in the presence of 4% cecal content. Degradation of the chitosan hydrogel beads in the presence of extracellular enzymes as compared with rat cecal and colonic enzymes indicates the potential of this multiparticulate system to serve as a carrier to deliver macromolecules specifically to the colon and can be offered as a substitute in vitro system for performing degradation studies. Studies demonstrated that orally administered chitosan hydrogel beads can be used effectively for the delivery of drug to the colon. PMID- 17915807 TI - Solventless photocurable film coating: evaluation of drug release, mechanical strength, and photostability. AB - A new solventless photocurable film-coating system was investigated in which nonpareil beads were coated in a mini-coating pan with liquid prepolymer (L) and powdered solid pore-forming agents (S) and cured by UV light. Release from the coating could by altered by changing the material, the number of layers, and the coating thickness. Immediate release of a blue dye contained in the nonpareils was obtained with sodium starch glycolate as a pore former that swelled the coating and yielded large pores; through these pores the dye quickly released while leaving behind the scaffold provided by the photocured prepolymer. Simple pore formers (lactose and sodium chloride) dissolved away without swelling and provided a more sustained release. The nature of the scaffold and pore structure of the coating were determined by simultaneously monitoring the release of sodium chloride from the coating and blue dye from the beads. At least 50% of the sodium chloride that was incorporated into the coating released before the dye released through the coating, except at an S/L ratio (ratio of the amount of solid pore forming agent to the volume of liquid prepolymer) of 2.4, where 40% of the sodium chloride was released before the release of dye. The coupling between dye release and pore formation was found to be dependent on the S/L ratio of the coating. Simulation based on percolation theory showed that the coupling of pore formation and dye release was higher when the variance in tortuosity was lower. The coating was photostable and could withstand normal handling stress. PMID- 17915808 TI - Rapidly dissolving repaglinide powders produced by the ultra-rapid freezing process. AB - The objective of the study was to produce rapidly dissolving formulations of the poorly water-soluble drug repaglinide using an innovative new technology, ultra rapid freezing (URF), and to investigate the influence of excipient type on repaglinide stability. Repaglinide compositions containing different types and levels of excipients and different drug potencies (50%-86%) were produced by the URF technology. Repaglinide/excipient solutions were frozen on a cryogenic substrate, collected, and lyophilized to form a dry powder. Surfactants, including sodium dodecyl sulfate, and alkalizing agents such as diethanolamine (DEA) and tromethamine (TRIS) were incorporated into the compositions. Forced degradation of repaglinide was conducted under stressed conditions (eg, elevated temperature, exposure to peroxide) to determine the stability of the drug in such environments. The solubility of repaglinide increased as a function of increasing pH; therefore, incorporation of an alkalizing agent into the URF formulations increased the drug's solubility. Drug instability resulted when the drug was exposed to pH values above 9.0. URF formulations containing alkalizing agents showed no degradation or spontaneous recrystallization in the formulation, indicating that increased stability was afforded by processing. URF processing created nanostructured drug/excipient particles with higher dissolution rates than were achieved for unprocessed drug. Alkalizing agents such as TRIS and DEA, present at levels of 25% to 33% wt/wt in the formulations, did not cause degradation of the drug when processed using URF. URF processing, therefore, yielded fast-dissolving formulations that were physically and chemically stable, resistant to alkali degradation or spontaneous recrystallization in the formulation. PMID- 17915809 TI - Design and evaluation of deformable talc agglomerates prepared by crystallo-co agglomeration technique for generating heterogeneous matrix. AB - The crystallo-co-agglomeration technique was used to design directly compressible and deformable agglomerates of talc containing the low-dose drug bromhexine hydrochloride (BXH). The process of agglomeration involved the use of dichloromethane as a good solvent and bridging liquid for BXH, water as a poor solvent, talc as diluent, and Tween 80 to aid dispersion of BXH and diluent into the poor solvent. Hydroxypropyl methylcellulose (50 cps) 4% wt/wt was used to impart the desired mechanical strength and polyethylene glycol 6000 5% wt/wt was used to impart the desired sphericity to the agglomerates. Clarity of the supernatant was considered an endpoint for completion of the agglomeration process. The drug-to-talc ratio in optimized batch 1 (BT1) and batch 2 (BT2) was kept at 1:15.66 and 1:24, respectively. The spherical agglomerates obtained were evaluated for topographic, micromeritic, mechanical, deformation, compressional, and drug release properties. The agglomeration yield and drug entrapment for both batches were above 94% wt/wt. Crushing strength and friability studies showed good handling qualities of agglomerates. Heckel plot studies showed low mean yield pressure and high tensile strength, indicating excellent compressibility and compactibility of agglomerates. Diametral and axial fracture of compacts showed deformation of agglomerates revealing formation of a heterogeneous compact. Drug release was sustained for 9 hours and 5 hours from BT1 and BT2, respectively, in 0.1N HCl. Hence, the crystallo-co-agglomeration technique can be successfully used for obtaining spherical, deformable, and directly compressible agglomerates, generating a heterogeneous matrix system and providing sustained drug release. PMID- 17915810 TI - Characterization of beta-lapachone and methylated beta-cyclodextrin solid-state systems. AB - The purpose of this research was to explore the utility of beta cyclodextrin (betaCD) and beta cyclodextrin derivatives (hydroxypropyl-beta-cyclodextrin [HPbetaCD], sulfobutylether-beta-CD [SBbetaCD], and a randomly methylated-beta-CD [RMbetaCD]) to form inclusion complexes with the antitumoral drug, beta-lapachone (betaLAP), in order to overcome the problem of its poor water solubility. RMbetaCD presented the highest efficiency for betaLAP solubilization and was selected to develop solid-state binary systems. Differential scanning calorimetry (DSC), X-ray powder diffractometry (XRPD), Fourier transform infrared (FTIR) and optical and scanning electron microscopy results suggest the formation of inclusion complexes by both freeze-drying and kneading techniques with a dramatic improvement in drug dissolution efficiency at 20-minute dissolution efficiency (DE(20-minute) 67.15% and 88.22%, respectively) against the drug (DE(20-minute) 27.11%) or the betaCD/drug physical mixture (DE(20-minute) 27.22%). However, the kneading method gives a highly crystalline material that together with the adequate drug dissolution profile make it the best procedure in obtaining inclusion complexes of RMbetaCD/betaLAP convenient for different applications of betaLAP. PMID- 17915811 TI - Development of a bladder instillation of the indoloquinone anticancer agent EO-9 using tert-butyl alcohol as lyophilization vehicle. AB - The purpose of this research was to develop a stable bladder instillation of EO-9 for the treatment of superficial bladder cancer. First, stability and dissolution studies were performed. Subsequently, the freeze-drying process was optimized by determination of the freeze-drying characteristics of the selected cosolvent/water system and differential scanning calorimetry analysis of the formulation solution. Furthermore, the influence of the freeze-drying process on crystallinity and morphology of the freeze-dried product was determined with x ray diffraction analysis and scanning electron microscopy, respectively. Subsequently, a reconstitution solution was developed. This study revealed that tert-butyl alcohol (TBA) can be used to both dramatically improve the solubility and stability of EO-9 and to shorten the freeze-drying cycle by increasing the sublimation rate. During freeze drying, 3 TBA crystals were found: TBA hydrate ice crystals, crystals of TBA hydrate, and a third crystal, probably composed of TBA hydrate crystals containing approximately 90% to 95% TBA. Furthermore, it was shown that crystallization of TBA hydrate was inhibited in the presence of both sodium bicarbonate (NaHCO3) and mannitol. Addition of an annealing step resulted in a minor increase in the crystallinity of the freeze-dried product and formation of the delta-polymorph of mannitol. A stable bladder instillation was obtained after reconstitution of the freeze-dried product (containing 8 mg of EO 9, 20 mg of NaHCO3, and 50 mg of mannitol per vial) to 20 mL with a reconstitution solution composed of propylene glycol/water for injection (WfI)/NaHCO3/sodium edetate 60%/40%/2%/0.02% vol/vol/wt/wt, followed by dilution with WfI to a final volume of 40 mL. PMID- 17915812 TI - In vitro dissolution method for evaluation of buprenorphine in situ gel formulation: a technical note. PMID- 17915813 TI - Study of coat quality of tablets coated by an on-line Supercell coater. AB - The aim of this study was to investigate the nature of Supercell coating, an on line tablet coater that employed a unique pattern of airflow. Tablets coated at different spray rates (4, 6, 8, 10, and 12 mL/min) were analyzed to investigate the influence of different wetting conditions on the quality of coats formed. Scanning electron micrographs showed that tablet coats formed at a spray rate of 4 mL/min consisted of spray-dried droplets that did not coalesce. At a spray rate of 6 mL/min, surface roughness was found to be lower than at the other spray rates, and the coat appeared smoothest, whereby droplets seemed fused together. At higher spray rates, the droplets appeared as branching arms and scale-like structures. This was attributed to the spread of spray droplets by the processing air and mass transfer of wet coating materials between tablets. Further tests showed that coats formed at higher spray rates had higher drug yield, drug uniformity, color uniformity, and density. However, the variability in coat thickness was increased due to the mass transfer of coats and dissolution of tablet core surfaces by the coating material. Since coats of different characteristics can be formed in Supercell coating, the choice of wetting conditions would depend on the type of coat required and the coating materials used. PMID- 17915814 TI - Influence of chitosan type on the properties of extruded pellets with low amount of microcrystalline cellulose. AB - The purpose of this research was to study the influence of type of chitosan with different molecular weights, ie, 190 and 419 kDa, on properties of pellets prepared by extrusion/spheronization. The formulations, consisting of acetaminophen as model drug, chitosan, microcrystalline cellulose (MCC), and dibasic calcium phosphate dihydrate with/without sodium alginate, were extruded using a twin-screw extruder and water as the granulating liquid. With 30% wt/wt MCC and no added sodium alginate, spherical pellets were produced containing low and high molecular weight chitosan at a maximum amount of 60% and 40% wt/wt, respectively. With sodium alginate (2.5% wt/wt), pellets with either type of chitosan (60% wt/wt), MCC (17.5% wt/wt), and acetaminophen (20% wt/wt) could be produced indicating an improved pellet-forming ability. Type and amount of chitosan and added sodium alginate affected physical properties of pellets including size, roundness, crushing force, and drug release. Low molecular weight chitosan produced pellets with higher mean diameter, sphericity, and crushing force. Additionally, the pellets made of low molecular weight chitosan and added sodium alginate showed faster drug release in 0.1 N HCl but had slower drug release in pH 7.4 phosphate buffer. This indicated that drug release from pellets could be modified by the molecular weight of chitosan. In conclusion, the molecular weight of chitosan had a major influence on formation, physical properties, and drug release from the obtained pellets. PMID- 17915815 TI - Comparison of the effect of tromethamine and polyvinylpyrrolidone on dissolution properties and analgesic effect of nimesulide. AB - The solubilizing and absorption enhancer properties towards nimesulide (ND) of tromethamine (Tris) and polyvinylpyrrolidone (PVP) have been investigated. Solid binary systems were prepared at various drug-polymer ratios by mixing or coprecipitation, characterized by differential scanning calorimetry, X-ray diffractometry, and Fourier transform infrared spectroscopy, and tested for dissolution behavior. Both carriers improved drug dissolution and their performance depended on concentration of the hydrophilic carrier in coprecipitates. Tris was more effective than PVP, despite the amorphizing power of PVP as revealed by solid state analyses. Complete drug amorphiztion was attained at 1:3 (wt/wt) drug:PVP, 25% (wt/wt) ND in PVP. According to thermal behavior of ND and Tris, ND-Tris systems present a eutectic behavior. The eutectic composition was 30% ND-70% Tris at approximately 129 degrees C. Amorphous ND-PVP and eutectic ND-Tris mixtures showed an improvement of 5.55 and 6.6 times of drug dissolution efficiency, respectively. In vivo experiments in mice demonstrated that administration of 50 mg/kg of drug coprecipitated with PVP or Tris resulted, respectively, in a 50% and 94% reduction of acetic acid-induced writhings in comparison with pure drug, which, instead, was statistically ineffective as compared with the control group. Moreover, the eutectic mixture of ND-Tris demonstrated antiwrithing potency 1.88 times higher than amorphous ND-PVP coprecipitate. Thus, the solubilizing power, dissolution-enhancing effect, and analgesic effect enhancer ability toward the drug make Tris particularly suitable for developing a reduced-dose, fast-release solid oral dosage form of nimesulide. PMID- 17915816 TI - An energy-based population-balance approach to model granule growth and breakage in high-shear wet granulation processes. AB - The trend in granule size distribution during the experiment closely followed the predicted model with an initial increase in the weight fraction of the larger granules. This increase was possibly due to extensive breakage of weaker granules and less extensive breakage, as if by attrition, of stronger granules, accompanied by the attachment of dry powder to the cracked surfaces. Eventually, larger granules experience increased impact energy and break. When excess binder is added and, higher volumes of powder reattach to the crack surface, more large granules form leading to granule overgrowth. This model highlights the importance of the probability of impact per unit time interval (ie, the rate of impact), the strength of the granules and the volume of powder that could attach to the cracked surface in high shear granulation processes where significant granule breakage is encountered. PMID- 17915817 TI - Artemisia arborescens L essential oil loaded beads: preparation and characterization. AB - The purpose of this work was to prepare sodium alginate beads as a device for the controlled release of essential oil for oral administration as an antiviral agent. Different formulations were prepared with sodium alginate as a natural polymer and calcium chloride or glutaraldehyde as a cross-linking agent. Loading capacities of between 86% and 100% were obtained in freshly prepared beads by changing exposure time to the cross-linking agent. Drying of the calcium alginate beads caused only a slight decrease in the loading efficiency. The surface morphology of the different bead formulations were studied using scanning electron microscopy (SEM). Stability studies over a 3-month period showed that glutaraldehyde reacted with some components of Artemisia arborescens L essential oil, changing its composition. Calcium alginate beads showed an in vitro controlled release of the essential oil for the investigated 24 hours, while the use of glutaraldehyde as a cross-linking agent was found not appropriate because of the interactions with azulene derivatives and the low degree of matrix cross linkage. PMID- 17915818 TI - A novel solid dosage form of rifampicin and isoniazid with improved functionality. AB - The aim of the present investigation was to develop a novel dosage form of rifampicin and isoniazid to minimize degradation of rifampicin in acidic medium and to modulate the release of rifampicin in the stomach and isoniazid in the intestine. Gastroretentive tablets of rifampicin (150 mg) were prepared by the wet granulation method using hydroxypropyl methylcellulose, calcium carbonate, and polyethylene glycol 4000. The granules and tablets of rifampicin were characterized. Hard gelatin capsules (size 4) containing a compacted mass of isoniazid (150 mg) and dicalcium phosphate (75 mg) were enteric coated. Two tablets of rifampicin and 1 capsule (size 4) of isoniazid were put into a hard gelatin capsule (size 00). The in vitro drug release and in vitro drug degradation studies were performed. Rifampicin was released over 4 hours by zero order kinetics from the novel dosage form. More than 90% of isoniazid was released in alkaline medium in 30 minutes. The results of dissolution studies with the US Pharmacopeia XXIII method revealed that a substantial amount of rifampicin was degraded from the immediate release capsule containing rifampicin and isoniazid powder owing to drug accumulation in the dissolution vessel and also to the presence of isoniazid. The degradation of rifampicin to 3-formyl rifampicin SV (3FRSV) was arrested (3.6%-4.8% degradation of rifampicin at 4 hours) because of the minimization of physical contact between the 2 drugs and controlled release of rifampicin in acidic medium in the modified Rossett-Rice apparatus. This study concludes that the problem of rifampicin degradation can be alleviated to a certain extent by this novel dosage form. PMID- 17915819 TI - Statistical evaluation of influence of viscosity and content of polymer on dipyridamole release from floating matrix tablets: a technical note. PMID- 17915821 TI - Effect of curing on water diffusivities in acrylate free films as measured via a sorption technique. AB - Studies were performed to investigate the effect of curing on the diffusion coefficients of water, as measured via the sorption technique, in acrylate polymeric films. The mathematical model selected for obtaining diffusion constants from the vapor-phase sorption studies was derived from the long-time Fourier equation used for diffusion into a planar sheet. For Eudragit NE films, the diffusion coefficients of water decreased continuously until a constant minimum value was reached. Diffusion coefficients in Eudragit RS films decreased initially but increased beyond 4 hours of curing at 70 degrees C and 90 degrees C. This latter result suggested the possible evaporation of plasticizer, which also results in a more dramatic increase in glass transition temperature with curing for the Eudragit RS free film in comparison to the Eudragit NE free film. Such loss of plasticizer could also lead to the formation of molecular-scale channels within the films, which would result in increased film permeability. To verify this proposed explanation, the amounts of triethyl citrate plasticizer in Eudragit RS free films were determined using Fourier-transform infrared spectrophotometry. An optimal curing condition was predicted for Eudragit NE and Eudragit RS films based upon the curing conditions at which a minimum value of the diffusion coefficient was reached. PMID- 17915820 TI - A novel fiber-optic photometer for in situ stability assessment of concentrated oil-in-water emulsions. AB - The purpose of this research was to evaluate a novel fiber-optic photometer for its ability to monitor physical instabilities occurring in concentrated emulsions during storage. For this, the fiber-optic photometer was used to measure transmission of oil-in-water emulsions stabilized with hypromellose (HPMC) as a function of oil volume fraction and droplet size distribution (DSD). To detect physical instabilities like creaming and coalescence, the transmissivity of the samples was studied at 2 different height levels over a certain period of time. The corresponding droplet size distributions were determined by laser diffraction with PIDS. Transmissivity was found to depend on the number of dispersed droplets and thus is sensitive to both the variation of phase volume fraction as well as the emulsions droplet size distribution. At constant DSD, light transmission decreased linearly with increasing oil content within a large interval of phase volume fractions from 0.01 to 0.3. At constant phase volume fraction, an increase in droplet size increased light transmission. Investigation of creaming on emulsions with different droplet size distributions showed changes in the initial delay times and creaming velocities. In contrast to creaming phenomenon coalescence can be identified by height independent changes of the transmissivity. In conclusion, transmissivity of oil-in-water emulsions observed by the novel fiber-optic photometer is sensitive to phase volume fraction, droplet size distribution, and thus can be used as a tool for stability studies on concentrated emulsions. PMID- 17915822 TI - Sodium alginate-magnesium aluminum silicate composite gels: characterization of flow behavior, microviscosity, and drug diffusivity. AB - The aims of the present study were to characterize the flow behavior and thixotropic properties of sodium alginate-magnesium aluminum silicate (SA-MAS) composite gels with various ratios of SA and MAS, and to investigate the drug diffusivity and microviscosity of the composite gels. Moreover, interaction of SA and MAS in the form of dry composite was examined by using Fourier Transform Infrared (FTIR), and a possible structure model of SA-MAS composite gel was illustrated. Incorporating MAS into the SA gels provided higher viscosity and changed the flow behavior from Newtonian to pseudoplastic with thixotropy. This was due to the formation of electrostatic force and intermolecular hydrogen bonding between SA and MAS, leading to a denser matrix structure of the composite gels. Increasing the content of MAS decreased the drug diffusivity but increased the microviscosity of the composite gels. The denser matrix structure of the composite gels had a higher tortuosity, resulting in slower drug diffusion through water-filled channels in the gels. This finding suggested that incorporating MAS into the SA gels could improve the flow behavior and sustain drug release from the gels because of the formation of a matrix structure between SA and MAS in the gels. PMID- 17915824 TI - Sintering of wax for controlling release from pellets. AB - The purpose of the present study was to investigate incorporation of hydrophobic (ie, waxy) material into pellets using a thermal sintering technique and to evaluate the pellets in vitro for controlled release. Pellets prepared by extrusion-spheronization technology were formulated with a water-soluble drug, microcrystalline cellulose, and carnauba wax. Powdered carnauba wax (4%-20%) prepared by grinding or by emulsification was studied with an attempt to retard the drug release. The inclusion of ground or emulsified carnauba wax did not sustain the release of theophylline for more than 3 hours. Matrix pellets of theophylline prepared with various concentrations of carnauba wax were sintered thermally at various times and temperatures. In vitro drug release profiles indicated an increase in drug release retardation with increasing carnauba wax concentration. Pellets prepared with ground wax showed a higher standard deviation than did those prepared with emulsified wax. There was incomplete release at the end of 12 hours for pellets prepared with 20% ground or emulsified wax. The sintering temperature and duration were optimized to allow for a sustained release lasting at least 12 hours. The optimized temperature and duration were found to be 100 degrees C and 140 seconds, respectively. The sintered pellets had a higher hydrophobicity than did the unsintered pellets. Scanning electron micrographs indicated that the carnauba wax moved internally, thereby increasing the surface area of wax within the pellets. PMID- 17915823 TI - Development and in vitro evaluation of an oral floating matrix tablet formulation of diltiazem hydrochloride. AB - The purpose of this research was to prepare a floating drug delivery system of diltiazem hydrochloride (DTZ). Floating matrix tablets of DTZ were developed to prolong gastric residence time and increase its bioavailability. Rapid gastrointestinal transit could result in incomplete drug release from the drug delivery system above the absorption zone leading to diminished efficacy of the administered dose. The tablets were prepared by direct compression technique, using polymers such as hydroxypropylmethylcellulose (HPMC, Methocel K100M CR), Compritol 888 ATO, alone or in combination and other standard excipients. Sodium bicarbonate was incorporated as a gas-generating agent. The effects of sodium bicarbonate and succinic acid on drug release profile and floating properties were investigated. A 3(2) factorial design was applied to systematically optimize the drug release profile. The amounts of Methocel K100M CR (X1) and Compritol 888 ATO (X2) were selected as independent variables. The time required for 50% (t50) and 85% (t85) drug dissolution were selected as dependent variables. The results of factorial design indicated that a high level of both Methocel K100M CR (X1) and Compritol 888 ATO (X2) favors the preparation of floating controlled release of DTZ tablets. Comparable release profiles between the commercial product and the designed system were obtained. The linear regression analysis and model fitting showed that all these formulations followed Korsmeyer and Peppas model, which had a higher value of correlation coefficient (r). While tablet hardness had little or no effect on the release kinetics and was found to be a determining factor with regards to the buoyancy of the tablets. PMID- 17915825 TI - Micromatricial metronidazole benzoate film as a local mucoadhesive delivery system for treatment of periodontal diseases. AB - The main objective of this study was to develop a local, oral mucoadhesive metronidazole benzoate (MET) delivery system that can be applied and removed by the patient for the treatment of periodontal diseases. Mucoadhesive micromatricial chitosan/poly(epsilon-caprolactone) (CH/PCL) films and chitosan films were prepared. Thermal behavior, morphology, and particle size measurements were used to evaluate the prepared films. The effect of different molar masses of CH and different ratios of medium Mwt molar mass chitosan (MCH):PCL on water absorption, in vitro bioadhesion, mechanical properties, and in vitro drug release was examined. In vivo performance of the selected formulation was also evaluated. Differential scanning calorimetry examination revealed that MET existed mainly in amorphous form. Under microscopic examination, PCL microparticles were homogeneously dispersed in the films. The use of different molar masses of CH and different ratios of (MCH):PCL affected the size of the entrapped particles. Addition of PCL significantly decreased percentage water uptake and bioadhesion force compared with pure CH film. With regard to mechanical properties, the 2-layered film containing 1:0.625 MCH:PCL had the best tensile properties. At fixed CH:PCL ratio (1:1.25), the slowest drug release was obtained from films containing high molar mass CH. On the other hand, the 2 layered film that consisted of 1:0.625 MCH:PCL had the slowest MET release. In vivo evaluation of the selected film revealed that metronidazole concentration in saliva over 6 hours ranged from 5 to 15 microg/mL, which was within and higher than the reported range of minimum inhibitory concentration for metronidazole. A significant in vitro/in vivo correlation under the adopted experimental conditions was obtained. PMID- 17915827 TI - Mucoadhesive bilayer tablets of propranolol hydrochloride. AB - The purpose of this research was to study mucoadhesive bilayer buccal tablets of propranolol hydrochloride using the bioadhesive polymers sodium alginate (Na alginate) and Carbopol 934P (CP) along with ethyl cellulose as an impermeable backing layer. The tablets were evaluated for weight variation, thickness, hardness, friability, surface pH, mucoadhesive strength, swelling index, in vitro drug release, ex vivo drug permeation, ex vivo mucoadhesion, and in vivo pharmacodynamics in rabbits. Tablets containing Na-alginate and CP in the ratio of 5:1 (F2) had the maximum percentage of in vitro drug release without disintegration in 12 hours. The swelling index was proportional to Na-alginate content and inversely proportional to CP content. The surface pH of all tablets was found to be satisfactory (7.0 +/- 1.5), close to neutral pH; hence, buccal cavity irritation should not occur with these tablets. The mechanism of drug release was found to be non-Fickian diffusion and followed zero-order kinetics. The formulation F4 was optimized based on good bioadhesive strength (28.9 +/- 0.99 g) and sustained in vitro drug permeation (68.65% +/- 3.69% for 12 hours). The behavior of formulation F4 was examined in human saliva, and both the drug and the buccal tablet were found to be stable. The formulation F4 was applied to rabbit oral mucosa for in vivo studies. The formulation inhibited isoprenaline induced tachycardia. The studies conducted in rabbits confirmed the sustained release as compared with intravenous administration. PMID- 17915826 TI - Compressed matrix core tablet as a quick/slow dual-component delivery system containing ibuprofen. AB - The purpose of the present research was to produce a quick/slow biphasic delivery system for ibuprofen. A dual-component tablet made of a sustained release tableted core and an immediate release tableted coat was prepared by direct compression. Both the core and the coat contained a model drug (ibuprofen). The sustained release effect was achieved with a polymer (hydroxypropyl methylcellulose [HPMC] or ethylcellulose) to modulate the release of the drug. The in vitro drug release profile from these tablets showed the desired biphasic release behavior: the ibuprofen contained in the fast releasing component was dissolved within 2 minutes, whereas the drug in the core tablet was released at different times (approximately 16 or >24 hours), depending on the composition of the matrix tablet. Based on the release kinetic parameters calculated, it can be concluded that the HPMC core was suitable for providing a constant and controlled release (zero order) for a long period of time. PMID- 17915828 TI - Influence of bulk and tapped density on the determination of the thermal conductivity of powders and blends. AB - The aim of this study was to evaluate a non-steady-state needle sensor to determine the thermal conductivity (lambda) of powders and their blends. It was investigated how lambda of different powders was influenced by (1) bulk vs tapped density, (2) moisture content of the powders, and (3) blending time of the powders. Different powders were evaluated: 2 lactose powders with different properties, a microcrystalline cellulose powder, a cornstarch powder, and 3 herbal extracts. The results show that the values of lambda are highly dependent on the bulk and tapped density of the powders. Bulk density measurements were generally not sensitive enough to detect the moisture content within a powder. The tapped density measurements were reliable and highly reproducible and could differentiate between the nature of a powder and the powder moisture content. Measurements of lambda were able to be used to monitor the powder blending process. To be able to use thermal conductivity measurements to characterize powder properties in quality control, the powder density must be defined because changes in density affect lambda. Using thermal conductivity as a measure for process analytical technology seems to be feasible and can add valuable information to the process under investigation. PMID- 17915829 TI - Development of polysaccharide gel-coated pellets for oral administration: swelling and release behavior of calcium pectinate gel. AB - The aim of this study was to examine the effect of pellet size, pectin type, pectin concentration, and dissolution medium on the swelling and drug release behavior of spherical pellets containing theophylline and coated with 2 different calcium pectinates, using a multi-level factorial design approach. The spherical pellets were prepared by an extrusion-spheronization method and then coated with calcium pectinate using the diffusion-controlled interfacial complexation technique, which provides a defect-free and uniform coating on solid cores. Theophylline release from the pellets was slowed by the application of the coatings. The time to release 50% of the payload (ie, T50) in an acidic medium was approximately 7 minutes from uncoated small pellets and was 55 minutes after an amidated calcium pectinate coat was applied; a comparable coat on large pellets showed a T50 of 93 minutes. Drug release profiles of dry coated pellets showed a lag time (all less than 20 minutes) when the gel coat hydrated and swelled, followed by a zero-order release. It was found that the release rate was controlled by the pellet size, pectin type, pectin concentration, and dissolution medium. PMID- 17915830 TI - Biomarkers, metabonomics, and drug development: can inborn errors of metabolism help in understanding drug toxicity? AB - Application of "omics" technology during drug discovery and development is rapidly evolving. This review evaluates the current status and future role of "metabonomics" as a tool in the drug development process to reduce the safety related attrition rates and bridge the gaps between preclinical and clinical, and clinical and market. Particularly, the review looks at the knowledge gap between the pharmaceutical industry and pediatric hospitals, where metabonomics has been successfully applied to screen and treat newborn babies with inborn errors of metabolism. An attempt has been made to relate the clinical pathology associated with inborn errors of metabolism with those of drug-induced pathology. It is proposed that extending the metabonomic biomarkers used in pediatric hospitals, as "advanced clinical chemistry" for preclinical and clinical drug development, is immediately warranted for better safety assessment of drug candidates. The latest advances in mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy should help replace the traditional approaches of laboratory clinical chemistry and move the safety evaluation of drug candidates into the new millennium. PMID- 17915831 TI - Inactivation of hepatic enzymes by inhalant nitrite--in vivo and in vitro studies. AB - We examined the effects of acute isobutyl nitrite (ISBN) exposure on the activity of several hepatic enzymes. Two strains of adult male mice (Balb/c and C57BL/6) were exposed to 900 ppm ISBN or ambient air for 45 minutes. The enzyme activity of hepatic cytochrome P450 (CYP)-mediated deethylation, glutathione S-transferase (GST), and carboxylesterase (CBE) was monitored through the substrates 3-cyano-7 ethoxycoumarin (CEC), 1-chloro-2,4-dinitrobenzene, and p-nitrophenyl acetate, respectively. Acute ISBN exposure led to a significant reduction in hepatic CYP mediated CEC deethylation, GST, and CBE activity in Balb/c mice (of 81.5%, 74.7%, and 25.2%, respectively, vs control mice, each at P < .05) when livers were harvested immediately after inhalant exposure. The corresponding decreases in C57BL/6 mice were smaller (with reductions of 21.8%, 18.8%, and 13.3%, respectively, each at P < .05). This enzyme activity, tested in C57BL/6 mice only, returned to control values after a 24-hour period of nonexposure. Follow-up mechanistic investigations using rat liver GST indicated that ISBN-mediated enzyme inactivation was not caused by its metabolites: inorganic nitrite ion (NO2 ) or nitric oxide. This inactivation could be prevented, but not reversed, by added glutathione, suggesting irreversible protein oxidation. Using different NO donors as comparative agents, we found that GST inactivation by ISBN was not associated with protein S-nitrosylation or disulfide formation, but with tyrosine nitration. Inhalant nitrite exposure, therefore, led to a significant reduction in hepatic enzyme activity in mice, possibly through tyrosine nitration of hepatic proteins. This effect raises the possibility of drug-drug metabolic interactions from inhalant nitrite abuse. However, determining the applicability of these findings to humans will require further study. PMID- 17915832 TI - Intellectual property policy in the pharmaceutical sciences: the effect of inappropriate patents and market exclusivity extensions on the health care system. AB - Though patents are effective tools for promoting innovation and protecting intellectual property in the pharmaceutical sciences, there has been growing concern about 2 important ways that patents in this field can have a negative effect on patient care and the practice of medicine. First, inventors can seek and receive patents on pharmaceutical products or research tools that stretch the statutory requirements for patenting. Second, patent holders in the pharmaceutical market can use legal loopholes or aspects of the patent registration system to extend exclusivity for inventions beyond what was anticipated by the Patent Act or subsequent legislation. The monopoly control bestowed by such inappropriate patents or manipulation of the patent system can limit options available to patients, increase the cost of health care delivery, and make cooperative research more difficult. In response, several different government and market-based efforts have emerged to promote more equitable patent policy in health care that encourages dissemination of ideas while still supporting the development of innovative products. PMID- 17915833 TI - The influence of market exclusivity on drug availability and medical innovations. AB - The interpretation and application of intellectual property laws is enormously complex in the pharmaceutical industry, with companies needing to obtain multiple patents to fully protect their innovations. While patents provide important incentives for biomedical innovation and economic growth, concern has been expressed over the growing number of patents, the granting of patents on basic research tools (eg, genetically engineered animals), and the possibility that these legal protections may ultimately inhibit scientific advancement. PMID- 17915835 TI - Silylenes: a unified picture of their stability, acid-base and spin properties, nucleophilicity, and electrophilicity via computational and conceptual density functional theory. AB - Conceptual DFT gives sharp definitions for many long-known, but rather vaguely defined chemical concepts. In this study DFT-based reactivity indices are applied to silylenes in order to elucidate the relationships among their properties: stability, acid-base, and spin properties, nucleophilicity and electrophilicity. On the basis of a detailed, comparative analysis of previously published data, it is shown that the properties of simple silylenes can be tuned by varying one single factor, the pi-electron donating ability of the substituents of the silicon atom leading to well-characterized and systematic changes in the stability/reactivity pattern of the molecule. In order to test the model a series of new compounds are studied: including CH3SiR (where R = CH3, NH2, OH and SH), Si(Si(CH3)3)2, Si(CF3)2 and benzo-, pyrido-, pyridazo-, and pyrimido-anellated 1,3,2lambda2-diazasiloles. PMID- 17915834 TI - Basics of US patents and the patent system. AB - The patent system plays an important role in stimulating the economy and advancing the quality of life in the United States. It serves as an incentive for innovation by giving inventors an exclusive right to their inventions for a limited period of time. It also increases and hastens the publication of useful knowledge by requiring inventors to disclose their invention to the public. Patents are particularly important in the pharmaceutical and biotechnology industries because they provide a mechanism by which the extremely high product development costs may be recouped. The United States Patent and Trademark Office acts as a gatekeeper in the patent system to prevent patents that do not meet the legal requirements from being thrust on the public. The legal requirements for obtaining a patent are discussed, particularly as they relate to pharmaceutical and biotechnological inventions. The process of examining an application for a patent is briefly described, along with some of the burdens faced by examiners when deciding the patentability of therapy-related inventions. PMID- 17915836 TI - Mechanism of thermal unimolecular decomposition of TNT (2,4,6-trinitrotoluene): a DFT study. AB - The widespread and long-term use of TNT has led to extensive study of its thermal and explosive properties. Although much research on the thermolysis of TNT and polynitro organic compounds has been undertaken, the kinetics and mechanism of the initiation and propagation reactions and their dependence on the temperature and pressure are unclear. Here, we report a comprehensive computational DFT investigation of the unimolecular adiabatic (thermal) decomposition of TNT. On the basis of previous experimental observations, we have postulated three possible pathways for TNT decomposition, keeping the aromatic ring intact, and calculated them at room temperature (298 K), 800, 900, 1500, 1700, and 2000 K and at the detonation temperature of 3500 K. Our calculations suggest that at relatively low temperatures, reaction of the methyl substituent on the ring (C-H alpha attack), leading to the formation of 2,4-dinitro-anthranil, is both kinetically and thermodynamically the most favorable pathway, while homolysis of the C-NO(2) bond is endergonic and kinetically less favorable. At approximately 1250-1500 K, the situation changes, and the C-NO(2) homolysis pathway dominates TNT decomposition. Rearrangement of the NO(2) moiety to ONO followed by O-NO homolysis is a thermodynamically more favorable pathway than the C-NO(2) homolysis pathway at room temperature and is the most exergonic pathway at high temperatures; however, at all temperatures, the C-NO(2) --> C-ONO rearrangement homolysis pathway is kinetically unfavorable as compared to the other two pathways. The computational temperature analysis we have performed sheds light on the pathway that might lead to a TNT explosion and on the temperature in which it becomes exergonic. The results appear to correlate closely with the experimentally derived shock wave detonation time (100-200 fs) for which only the C-NO(2) homolysis pathway is kinetically accessible. PMID- 17915837 TI - A density functional study of the various forms of UN4O12 containing uranyl nitrate. AB - In this paper we report the computational results of a density functional study of 73 UN4O12 isomers containing uranyl nitrate, UO2(NO3)2, as a component. The isomers are grouped into three categories and 19 types. Forty-four isomers of 14 types are dinitrogen tetroxide adducts of uranyl nitrate, UO2(NO3)2.N2O4, 22 are nitrosonium salt adducts of uranyl nitrate, NO+UO2(NO3)3-, NO+UO2(NO3)2O(NO2)-, NO+UO2(NO3)2(ONOO)-, or (NO+)2UO2(NO3)2O22-, and 7 are bis(nitrogen dioxide) adducts of uranyl nitrate, UO2(NO3)2.2NO2. The 22 most stable isomers in solution, representing the 20 most stable gas-phase isomers, were selected for analysis. Of these selected structures only two categories and six types were represented. Structures, frequencies, gas-phase and solution energetics, atomic charges, dipole moments, and the bonding within the N2O4 unit and between NO+ and UO2(NO3)3- components have been analyzed in detail. On the basis of relative Gibbs free energy calculations five isomers (the N2O4 adducts a1, a2, and a3 and the nitrosonium salts b1 and b2) were identified as strong candidates to exist and possibly predominate in the gas phase, with a1 and a2 being the strongest candidates. Similarly, four isomers (a6, a5, a8, and a1, all of them N2O4 adducts) were identified as strong candidates to exist and possibly predominate in a nonaqueous solution of nitromethane/dinitrogen tetroxide. Of these, a6 was determined to be the most likely candidate to predominate in solution. The possibility of dissociation in solution has been addressed briefly. In addition, computational evidence for the existence of four new N2O4 isomers 20, 22, 27, and 28 in both the gas and the solution phases is presented for the first time. PMID- 17915838 TI - Photophysics of water soluble perylene diimides in surfactant solutions. AB - Our previous photophysical study of water soluble perylene diimides (WS-PDIs) has suggested that WS-PDIs are present in aqueous solution in partially aggregated form (Tang, T. J.; Qu, J. Q.; Mullen, K.; Webber, S. E. Langmuir 2006, 22, 7610 7616) In this article we present a study of the effect of surfactants (dodecyltrimethylammonium chloride (DTAC), sodium dodecyl sulfate (SDS)) on the photophysics of WS-PDIs. Adding surfactant to WS-PDI solutions is accompanied by their increased fluorescence quantum yield and lifetime and a more structured absorption spectra. We are able to demonstrate that above the surfactant critical micelle concentration (cmc) the WS-PDI moieties are molecularly dispersed and isolated from each other. Our findings are consistent with the existence of weakly interacting aggregates of WS-PDIs in pure water, which can be broken up by surfactants even below the cmc, although we cannot rule out that the observed photophysical changes arise from modifying the local environment of molecularly solubilized WS-PDIs (e.g., local polarity or modification of the molecular planarity). PMID- 17915839 TI - Connecting structure to infrared spectra of molecular and autodissociated HCl- water aggregates. AB - The properties of perdeuterated HCl(H2O)n aggregates with n=1, 2, ..., 6 water molecules are studied by means of ab initio molecular dynamics simulations. The specific focus is on the phenomenon of autodissociation of the acid HCl as a function of the microsolvation environment size. The calculations provide a basis for characterization in terms of autodissociation energetics as well as in terms of the impact of thermal fluctuations on structure including proton fluxionality and in terms of anharmonic infrared vibrational spectra. Structure stabilization is dominated by strong hydrogen bonds resulting in distinct topologies, which, in turn, heavily influence acid dissociation. The latter is favored for the first time when n = 4. In that case, three hydrogen bonds can be donated toward the chlorine while at the same time a hydronium core is perfectly solvated according to the eigencomplex motif. Hydrogen-bonding interactions between DCl and its solvating molecules affect the dynamical behavior of the D-Cl bond significantly. This can be seen by the onset of fluxionality and an emerging tendency toward proton transfer for the larger clusters. Connecting IR spectra to structural information is possible by exploiting the following observations. Zwitterionic species show characteristic differences in the hydronium region, whereas the D-Cl stretching regime is useful to distinguish neutral aggregates. Furthermore, in the case of fluxional protons large-amplitude motion leads to characteristic band shifts and significant band broadening effects. PMID- 17915840 TI - A photochemical mechanism for homochirogenesis. AB - Theoretical analysis of one-step and multiple-step photoreactions initiated with circularly polarized light shows that the enantiomeric excess of a chiral reactant approaches +/- 1 as the amount of unreacted reactant approaches 0. The final product never has a large enantiomeric excess at any stage of its formation and slowly decreases to 0 at the completion of the reaction. For multiple-step reactions the behavior of the intermediate photoproducts is much more interesting. During certain stages of the overall reaction both the size of the enantiomer excess and the amount of a given intermediate photoproduct are large. Furthermore, the sign of the enantiomeric excess of an intermediate may change during the course of the reaction. Multiple-step photoreactions initiated with circularly polarized light may be a method by which the exogenous and endogenous synthesis of optically active molecules occurred in the prebiotic universe. PMID- 17915841 TI - Theoretical study of the fragmentation pathways of norbornane in its doubly ionized ground state. AB - The potential energy surface of norbornane in its dicationic singlet ground state has been investigated in detail using density functional theory along with the nonlocal hybrid and gradient-corrected Becke three-parameter Lee-Yang-Parr functional (B3LYP) and the cc-pVDZ basis set. For the sake of more quantitative insight into the chemical reactions induced by double ionization of norbornane, this study was supplemented by a calculation of basic thermodynamic state functions coupled to a focal point analysis of energy differences obtained using correlation treatments and basis sets of improving quality, enabling an extrapolation of these energy differences at the CCSD(T) level in the limit of an asymptotically complete (cc-pV infinity Z) basis set. Our results demonstrate the likelihood of an ultrafast intramolecular rearrangement of the saturated hydrocarbon cage after a sudden removal of two electrons into a kinetically metastable five-membered cyclic C5H8+-CH+-CH3 intermediate, prior to a Coulomb explosion into C5H7+=CH2 and CH3+ fragments, which might explain a tremendous rise of electron-impact (e, 2e) ionization cross sections at electron binding energies around the double-ionization threshold. The first step is straightforward and strongly exothermic (DeltaH298 = -114.0 kcal mol-1). The second step is also exothermic (DeltaH298 = -10.2 kcal mol-1) but requires an activation enthalpy (DeltaH298) of 39.7 kcal/mol. The various factors governing the structure of this intermediate, such as electrostatic interactions, inductive effects, cyclic strains, and methylenic hyperconjugation interactions, are discussed in detail. PMID- 17915842 TI - Hyperfine coupling in methyl radical isotopomers. AB - The hyperfine coupling constants (hfcs) of two methyl radical isotopomers, CH2Mu and CD2Mu, have been measured over a wide range of temperature in ketene and ketene-d2, from which the radicals were generated. The magnitudes of the hfcs of these muoniated methyl radical isotopomers are larger than those of CH3 and CD3 due to larger zero-point energy in the out-of-plane bending mode. In contrast to CH3 and CD3, where the coupling constants become smaller with increasing temperature, the negative hfcs of the muoniated radicals were found to increase in magnitude (become more negative) with temperature, passing through a maximum near the boiling point of ketene. This behavior is attributed to a solvent induced change in the force constant of the out-of-plane bending mode. The opposite temperature effect known for CH3 and CD3 is explained by excitation of the low frequency out-of-plane bending mode. This effect is much smaller in the muoniated radicals, where the vibrational frequency is significantly higher due to the light mass of muonium; consequently, the solvent effect dominates at low temperatures. PMID- 17915843 TI - Interpretation of the photoelectron spectra of FeS(2)(-) by a multiconfiguration computational approach. AB - The ground states of FeS(2) and FeS(2)(-), and several low-lying excited electronic states of FeS(2) that are responsible for the FeS(2)(-) photoelectron spectrum, are calculated. At the B3LYP level an open, quasi-linear [SFeS](-) conformation is found as the most stable structure, which is confirmed at the ab initio CASPT2 computational level. Both the neutral and the anionic unsaturated complexes possess high-spin electronic ground states. For the first time a complete assignment of the photoelectron spectrum of FeS(2)(-) is proposed. The lowest energy band in this spectrum is ascribed to an electron detachment from the two highest-lying 3dpi antibonding orbitals (with respect to the iron-sulfur bonding) of iron. The next-lowest experimental band corresponds to an electron removal from nonbonding, nearly pure sulfur orbitals. The two highest bands in the spectra are assigned as electron detachments from pi and sigma bonding mainly sulfur orbitals. PMID- 17915844 TI - Hydrogen bonding mediated by key orbital interactions determines hydration enthalpy differences of phosphate water clusters. AB - Electronic structure calculations have been carried out to provide a molecular interpretation for dihydrogen phosphate stability in water relative to that of metaphosphate. Specifically, hydration enthalpies of biologically important metaphosphate and dihydrogen phosphate with one to three waters have been computed with second-order Moller-Plesset perturbation and density functional theory (B3LYP) with up to the aug-cc-pvtz basis set and compared to experiment. The inclusion of basis set superposition error corrections and supplemental diffuse functions are necessary to predict hydration enthalpies within experimental uncertainty. Natural bond orbital analysis is used to rationalize underlying hydrogen bond configurations and key orbital interactions responsible for the experimentally reported difference in hydration enthalpies between metaphosphate and dihydrogen phosphate. In general, dihydrogen phosphate forms stronger hydrogen bonds compared to metaphosphate due to a greater charge transfer or enhanced orbital overlap between the phosphoryl oxygen lone pairs, n(O), and the antibonding O-H bond of water. Intramolecular distal lone pair repulsion with the donor n(O) orbital of dihydrogen phosphate distorts symmetric conformations, which improves n(O) and sigma*(O-H) overlap and ultimately the hydrogen bond strength. Unlike metaphosphate, water complexed to dihydrogen phosphate can serve as both a hydrogen bond donor and a hydrogen bond acceptor, which results in cooperative charge transfer and a reduction of the energy gap between n(O) and sigma*(O-H), leading to stronger hydrogen bonds. This study offers insight into how orbital interactions mediate hydrogen bond strengths with potential implications on the understanding of the kinetics and mechanism in enzymatic phosphoryl transfer reactions. PMID- 17915845 TI - Static dipole polarizability of ytterbium. AB - The static dipole polarizability of the ground state ytterbium atom is calculated using full and approximate relativistic ab initio methods. Our recommended polarizability of 143 au is consistent with experimental atomic spectral data. The corresponding van der Waals coefficient C6 of Yb2, derived using Pade approximants, is 2062 (200) au. PMID- 17915846 TI - Hydrolysis of N-sulfinylamines and isocyanates: a computational comparison. AB - A comparative study of the concerted hydrolysis of H-, CF3-, CH3-, and Ph substituted N-sulfinylamines (R-NSO) and isocyanates (R-NCO) was performed using B3LYP/6-31+G(2d,2p). The "two-water-molecule" model was found to be sufficient for a proper description of the hydrolysis reaction for both classes of compounds. Despite their overall similar reactivity, N-sulfinylamines react across both the N=S and the S=O bonds, whereas isocyanates hydrolyze predominantly through the N=C bond, in agreement with the proton affinities of the nitrogen and oxygen atoms. The charges on sulfur (N=S=O) and carbon (N=C=O) were found to correlate well with the activation enthalpy for hydrolysis. While the reactivity of an N-sulfinylamine toward water increases with increasing electron-withdrawing ability of the substituent, the substituent effect on isocyanates becomes notable only in the presence of the strongly electron withdrawing CF3 group. PMID- 17915847 TI - Stability, reactivity, and aromaticity of compounds of a multivalent superatom. AB - In this article, we analyze the stability, reactivity, and possible aromatic behavior of two recently reported clusters (Reveles, J. U.; Khanna, S. N.; Roach, P. J.; Castleman, A. W., Jr. Proc. Natl. Acad. Sci. 2006, 103, 18405), viz., Al(7)C(-) and Al(7)O(-) in the light of the principles of the maximum hardness and minimum electrophilicity as well as the nucleus-independent chemical shift values. Stability of these clusters in the context of addition/removal of an electron or an Al atom is now clearly understood. PMID- 17915849 TI - Pharmacomodulations around the 4-oxo-1,4-dihydroquinoline-3-carboxamides, a class of potent CB2-selective cannabinoid receptor ligands: consequences in receptor affinity and functionality. AB - CB2 receptor selective ligands are becoming increasingly attractive drugs due to the potential role of this receptor in several physiopathological processes. Thus, the development of our previously described series of 4-oxo-1,4 dihydroquinoline-3-carboxamides was pursued with the aim to further characterize the structure-affinity and structure-functionality relationships of these derivatives. The influence of the side chain was investigated by synthesizing compounds bearing various carboxamido and keto substituents. On the other hand, the role of the quinoline central scaffold was studied by synthesizing several 6 , 7-, or 8-chloro-4-oxo-1,4-dihydroquinolines, as well as 4-oxo-1,4 dihydronaphthyridine and 4-oxo-1,4-dihydrocinnoline derivatives. The effect of these modifications on the affinity and functionality at the CB2 receptor was studied and allowed for the characterization of new selective CB2 receptor ligands. PMID- 17915848 TI - Small molecule inhibitors of integrin alpha2beta1. AB - Interactions between the integrin, alpha2beta1, and extracellular matrix (ECM), particularly collagen, play a pivotal role in platelet adhesion and thrombus formation. Platelets interact with collagen in the subendothelial matrix that is exposed by vascular damage. To evaluate the potential of alpha2beta1 inhibitors for anticancer and antithrombotic applications, we have developed a series of small molecule inhibitors of this integrin based on a prolyl-2,3-diaminopropionic acid (DAP) scaffold using solid-phase parallel synthesis. A benzenesulfonamide substituent at the N-terminus of the dipepetide and a benzyl urea at the DAP side chain resulted in tight and highly selective inhibition of alpha2beta1-mediated adhesion of human platelets and other cells to collagen. PMID- 17915851 TI - 3-substituted 7-phenyl-pyrroloquinolinones show potent cytotoxic activity in human cancer cell lines. AB - A novel series of 3-alkyl-substituted 7-phenyl-3H-pyrrolo[3,2-f]quinolin-9-ones (7-PPyQs) was synthesized with the aim to optimize the cytotoxic activity of recently identified PPyQs, promising inhibitors of tubulin polymerization. All compounds inhibited the growth of 11 human tumor cell lines at submicromolar concentrations as well as two human resistant cancer sublines, A549-T12 and A549 T24. FACS analysis indicated that all compounds caused significant arrest of the A549 cell cycle in G2/M phase at 0.1 and 1 muM and a good correlation between the cytotoxicity IC50 and their ability to block the cell cycle was observed. PMID- 17915850 TI - NMR structural studies of the supramolecular adducts between a liver cytosolic bile acid binding protein and gadolinium(III)-chelates bearing bile acids residues: molecular determinants of the binding of a hepatospecific magnetic resonance imaging contrast agent. AB - The binding affinities of a selected series of Gd(III) chelates bearing bile acid residues, potential hepatospecific MRI contrast agents, to a liver cytosolic bile acid transporter, have been determined through relaxivity measurements. The Ln(III) complexes of compound 1 were selected for further NMR structural analysis aimed at assessing the molecular determinants of binding. A number of NMR experiments have been carried out on the bile acid-like adduct, using both diamagnetic Y(III) and paramagnetic Gd(III) complexes, bound to a liver bile acid binding protein. The identified protein "hot spots" defined a single binding site located at the protein portal region. The presented findings will serve in a medicinal chemistry approach for the design of hepatocytes-selective gadolinium chelates for liver malignancies detection. PMID- 17915852 TI - Structures of human monoamine oxidase B complexes with selective noncovalent inhibitors: safinamide and coumarin analogs. AB - Structures of human monoamine oxidase B (MAO B) in complex with safinamide and two coumarin derivatives, all sharing a common benzyloxy substituent, were determined by X-ray crystallography. These compounds competitively inhibit MAO B with Ki values in the 0.1-0.5 microM range that are 30-700-fold lower than those observed with MAO A. The inhibitors bind noncovalently to MAO B, occupying both the entrance and the substrate cavities and showing a similarly oriented benzyloxy substituent. PMID- 17915853 TI - CYP2C9 structure-metabolism relationships: substrates, inhibitors, and metabolites. AB - The cytochrome P450 (CYP) family is composed of monooxygenases, which mediate the metabolism of xenobiotics and endogenous compounds. The characterization of the interactions between these enzymes and candidate drugs is an important part of the drug discovery process. CYP2C9, one isoform of the CYPs, mediates the oxidation of several important drugs. The aim of this work is to investigate the possibility to study inhibition and substrate interactions with CYP2C9, using docking and the site of metabolism predictions. The model compounds used for the study were the COX-2 inhibitor celecoxib and a series of 13 analogues known to be metabolized by CYP2C9. The results obtained using the two methods gave valuable information about important interactions of inhibitors and substrates with CYP2C9. The two methods could be used to predict the site of metabolism and to determine the productive docking pose for each compound. These predictions were verified by metabolite identification using LC/MS/MS after incubation with recombinant CYP2C9. PMID- 17915854 TI - Cyclooxygenase-2 inhibitors. 1,5-diarylpyrrol-3-acetic esters with enhanced inhibitory activity toward cyclooxygenase-2 and improved cyclooxygenase 2/cyclooxygenase-1 selectivity. AB - The important role of cyclooxygenase-2 (COX-2) in the pathogenesis of inflammation and side effect limitations of current COX-2 inhibitor drugs illustrates a need for the design of new compounds based on alternative structural templates. We previously reported a set of substituted 1,5 diarylpyrrole derivatives, along with their inhibitory activity toward COX enzymes. Several compounds proved to be highly selective COX-2 inhibitors and their affinity data were rationalized through docking simulations. In this paper, we describe the synthesis of new 1,5-diarylpyrrole derivatives that were assayed for their in vitro inhibitory effects toward COX isozymes. Among them, the ethyl 2-methyl-5-[4-(methylsulfonyl)phenyl]-1-[3-fluorophenyl]-1H-pyrrol-3-acetate (1d), which was the most potent and COX-2 selective compound, also showed a very interesting in vivo anti-inflammatory and analgesic activity, laying the foundations for developing new lead compounds that could be effective agents in the armamentarium for the management of inflammation and pain. PMID- 17915857 TI - Isolation of a pentadentate ligand and stepwise synthesis, structures, and magnetic properties of a new family of homo- and heterotrinuclear complexes. AB - A neutral pentadentate ligand, di(pyrazolecarbimido)amine (Hdcadpz), and its adduct with HClO4, [H2dcadpz]+[ClO4]-, were for the first time isolated from our previously reported [Cu3(dcadpz)2(Hpz)2(ClO4)2](ClO4)2.H2O by the use of (NH4)2S to remove the CuII ions and characterized by IR, EA, UV, NMR, MS, and X-ray crystallography. Reactions of copper(II) or nickel(II) nitrate with Hdcadpz in a 1:2 molar ratio generated two mononuclear precursors of [Cu(dcadpz)2] (1) and [Ni(dcadpz)2].2/3DMF (2). Furthermore, three new linear homo- and heterotrinuclear complexes of the same motif [M{M'(dcadpz)2}M] (M=CoII, NiII, M'=CuII, NiII), [{Co(pdm)}2{Cu(dcadpz)2}](NO3)4 (3), [{Ni(pdm)}2{Cu(dcadpz)2}](NO3)4 (4), and [{Ni(MeOH)(H2O)2}2{Ni(dcadpz)2}](NO3)4 (5), were synthesized from these two precursors (pdm=2,6-pyridinedimethanol) and characterized by X-ray crystallography. Magnetic studies show that the central Cu(dcadpz)2 motif is antiferromagnetically coupled with both the terminal Co(II) atoms via the dcadpz- ligand in 3 with a J value of -5.27 cm(-1) and ferromagnetically coupled with both the terminal Ni(II) atoms in 4 with a J value of 2.50 cm(-1), while 5 behaves only as a Curie paramagnet between 2 and 300 K due to the diamagnetic character of the central square-planar Ni(II) atom. PMID- 17915856 TI - Chemical data mining of the NCI human tumor cell line database. AB - The NCI Developmental Therapeutics Program Human Tumor cell line data set is a publicly available database that contains cellular assay screening data for over 40 000 compounds tested in 60 human tumor cell lines. The database also contains microarray assay gene expression data for the cell lines, and so it provides an excellent information resource particularly for testing data mining methods that bridge chemical, biological, and genomic information. In this paper we describe a formal knowledge discovery approach to characterizing and data mining this set and report the results of some of our initial experiments in mining the set from a chemoinformatics perspective. PMID- 17915858 TI - A 3D porous cobalt-organic framework exhibiting spin-canted antiferromagnetism and field-induced spin-flop transition. AB - Two 3D cobalt-organic frameworks formulated as [Co3(2,4-pydc)2(micro3 OH)2]n.5nH2O (1) and [Co3(2,4-pydc)2(micro3-OH)2(H2O)]n.7nH2O (2) (2,4 pydc=pyridine-2,4-dicarboxylate) have been hydrothermally synthesized and characterized. Both compounds 1 and 2 exhibit the 3D porous frameworks with hydroxyl-bridged metal Delta-chains. However, in comparison with only two crystallographically independent CoII ions in a unit of 2, three crystallographically independent CoII ions are found in an asymmetric unit of 1, where their Delta-chains are constructed by two types of vertexes sharing quadrangles formed via edge-sharing triangles. Magnetic studies show that 1 exhibits spin-canted antiferomagnetism and a field-induced spin-flop transition while 2 behaves as a normal antiferromagnet. The magnetic properties are largely retained by the porous frameworks of dehydrated 1 and 2 compounds. Gas adsorption measurements indicate that both the dehydrated compounds absorb H2 into their pores. PMID- 17915859 TI - In situ ligand reactions under hydrothermal conditions afford a novel zinc substituted polyoxovanadate dimer. AB - A novel polyoxovanadate (POV), [Zn(2,2' bpy)3]4[(ppz){{Zn(tepa)}2ZnAs8V13O41(H2O)}2][As8V14O42(0.5H2O)]2.4H2O (1, 2,2' bpy = 2,2'-bipyridine, ppz = piperazine, tepa = tetraethylenepentamine), has been hydrothermally synthesized and structurally characterized by elemental analysis, IR spectra, thermogravimetric analysis, magnetic measurement, and single-crystal X-ray diffraction analysis. Crystal data for 1: orthorhombic, Aba2, a = 28.3414(14) A, b = 27.9995(16) A, c = 41.5819(16) A, V = 32997(3) A3, Z = 4, rho = 2.264 mg.cm-3. X-ray structure analysis shows that the structure of 1 consists of one unique zinc-substituted POV dimer and two POV monomers. Interestingly, the formation of the zinc-substituted POV dimer involves two new in situ ligand reactions under hydrothermal conditions, the intermolecular and intramolecular deamination coupling reactions of dien. Magnetization measurement revealed the presence of antiferromagnetic interaction between V(IV) ions in 1. PMID- 17915860 TI - Structures and physical properties of new semiconducting polyselenides Ba2CudeltaAg4-deltaSe5 with unprecedented linear Se(3)4- units. AB - The title compounds were prepared from the elements in evacuated silica tubes at 650 degrees C, followed by slow cooling. Ba2Ag4Se5 forms a new structure type, space group C2/m, with a=16.189(2) A, b=4.5528(6) A, c=9.2500(1) A, beta=124.572(3) degrees, and V=561.4(1) A3 (Z=2). A maximum of 44% of the Ag atoms may be replaced with Cu atoms without changing the structure type. The crystal structure is composed of Ag4Se(5)4- layers, interconnected via the Ba2+ cations. The Ag atoms show irregular [3+1] coordination by the Se atoms, and the Ba atoms are located in capped square antiprisms formed by Se atoms. Most intriguing is the unprecedented occurrence of linear Se(3)4- units. According to the formulation (Ba2+)2(Ag+)4Se(3)4-(Se2-)2, this selenide is electron-precise with eight positive charges equalizing the eight negative charges. Electronic structure calculations indicated the presence of a band gap, as was experimentally confirmed: the electrical conductivity measurement revealed a gap of 0.6 eV for Ba2CuAg3Se5. PMID- 17915861 TI - Tuneable intramolecular intermetallic interactions as a new tool for programming linear heterometallic 4f-4f complexes. AB - Statistical mechanics predicts that the design of pure organized heteropolymetallic chains of metal ions bound to linear receptors depends on controlled deviations from the mixing rule DeltaE(MiMj) = 1/2 (DeltaE(MiMi) + DeltaE(MjMj)), whereby DeltaE(MiMj) is the intramolecular intermetallic interaction between neighboring metal i and metal j along the receptor. A thorough investigation of linear polymetallic trivalent lanthanide triple stranded helicates shows that such deviations are amplified by an increase in the nuclearity of the final complexes and are thus easily evidenced in the tetranuclear heterobimetallic helicates [La(4-y)Lu(y)(L6)3](12+) (y = 0-4). The chemical and physical origins of this unprecedented behavior are discussed together with its practical consequences for programming pure heteropolymetallic 4f-4f complexes. PMID- 17915862 TI - Dizinc enzyme model/complexometric indicator pairs in indicator displacement assays for inorganic phosphates under physiological conditions. AB - A dizinc phosphohydrolase enzyme model complex employing the dinucleating ligand 2,6-bis-[(bis-pyridin-2-ylmethyl-amino)methyl]-4-methylphenol (L1) was tested for binding to a series of 11 commercially available complexometric indicators in aqueous N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) buffer at pH 7.4, with the aim of determining the applicability of these indicators in indicator displacement assays (IDAs) under physiological conditions. Dissociation constants (Kd) were determined for 11 indicator-Zn2L1 complexes, spanning 2 orders of magnitude from 2.8 x 10(-4) M (alizarin red S) to 2.7 x 10(-6) M (bromo pyrogallol red). Phosphate and pyrophosphate were tested for their ability to displace bound indicator and produce a detectable colorimetric response. Three indicators (bromo pyrogallol red, mordant blue 9, and zincon) complex to Zn2L1 to form an indicator displacement assay selective for pyrophosphate over phosphate. Because selection of an indicator/analyte pair having appropriate relative Kd values is critical for their successful application in IDAs, the binding data for these 11 indicators should assist their extension to IDAs for other analytes. PMID- 17915863 TI - Photochemical synthesis of mer-[Re(bpy)(CO)3Cl]. AB - The mer isomer of Re(bpy)(CO)3Cl (bpy = 2,2'-bipyridine) has been synthesized photochemically and isolated for the first time. The mer complex is a useful starting material for the synthesis of cis,cis-[Re(bpy)(CO)2(L)Cl]-type complexes. PMID- 17915864 TI - Switching in molecular transport junctions: polarization response. AB - We discuss several proposed explanations for the switching and negative differential resistance (NDR) behavior seen in some molecular junctions. Several theoretical models are discussed, and we present results of electronic structure calculations on a series of substituted oligo(phenylene ethynylene) molecules. It is shown that a previously proposed polaron model is successful in predicting NDR behavior, and the model is elaborated with image charge effects and parameters from electronic structure calculations. This model now incorporates substituent effects and includes the effects of conformational change, charging, and image charge stabilization. PMID- 17915866 TI - Structural requirements and reaction pathways in dimethyl ether combustion catalyzed by supported Pt clusters. AB - The identity and reversibility of the elementary steps required for catalytic combustion of dimethyl ether (DME) on Pt clusters were determined by combining isotopic and kinetic analyses with density functional theory estimates of reaction energies and activation barriers to probe the lowest energy paths. Reaction rates are limited by C-H bond activation in DME molecules adsorbed on surfaces of Pt clusters containing chemisorbed oxygen atoms at near-saturation coverages. Reaction energies and activation barriers for C-H bond activation in DME to form methoxymethyl and hydroxyl surface intermediates show that this step is more favorable than the activation of C-O bonds to form two methoxides, consistent with measured rates and kinetic isotope effects. This kinetic preference is driven by the greater stability of the CH3OCH2* and OH* intermediates relative to chemisorbed methoxides. Experimental activation barriers on Pt clusters agree with density functional theory (DFT)-derived barriers on oxygen-covered Pt(111). Measured DME turnover rates increased with increasing DME pressure, but decreased as the O2 pressure increased, because vacancies (*) on Pt surfaces nearly saturated with chemisorbed oxygen are required for DME chemisorption. DFT calculations show that although these surface vacancies are required, higher oxygen coverages lead to lower C-H activation barriers, because the basicity of oxygen adatoms increases with coverage and they become more effective in hydrogen abstraction from DME. Water inhibits reaction rates via quasi-equilibrated adsorption on vacancy sites, consistent with DFT results indicating that water binds more strongly than DME on vacancies. These conclusions are consistent with the measured kinetic response of combustion rates to DME, O2, and H2O, with H/D kinetic isotope effects, and with the absence of isotopic scrambling in reactants containing isotopic mixtures of 18O2-16O2 or 12CH3O12CH3-13CH3O13CH3. Turnover rates increased with Pt cluster size, because small clusters, with more coordinatively unsaturated surface atoms, bind oxygen atoms more strongly than larger clusters and exhibit lower steady-state vacancy concentrations and a consequently smaller number of adsorbed DME intermediates involved in kinetically relevant steps. These effects of cluster size and metal oxygen bond energies on reactivity are ubiquitous in oxidation reactions requiring vacancies on surfaces nearly saturated with intermediates derived from O2. PMID- 17915865 TI - Charge transport parameters of the pentathienoacene crystal. AB - Pentathienoacene, the thiophene equivalent of pentacene, is one of the latest additions to the family of organic crystal semiconductors with a great potential for use in thin film transistors. By using density functional theory and gas phase ultraviolet photoelectron spectroscopy, we investigate the microscopic charge transport parameters of the pentathienoacene crystal. We find that the valence band exhibits a stronger dispersion than those in the pentacene and rubrene single crystals with marked uniaxial characteristics within the molecular layer due to the presence of one-dimensional pi-stacks; a small hole effective mass is also found along the direction perpendicular to the molecular layers. In the conduction band, strong intermolecular sulfur-sulfur interactions give rise to a significant interstack electronic coupling whereas the intrastack dispersion is greatly reduced. The intramolecular vibronic coupling (reorganization energy) is stronger than that in pentacene but comparable to that in sexithiophene; it is larger for holes than for electrons, as a result of low-frequency modes induced by the sulfur atoms. The polarization energy is large, but its effect on the vibronic coupling remains small. Charge transport is discussed in the framework of both band and hopping models. PMID- 17915867 TI - Quantitative inhibitor fingerprinting of metalloproteases using small molecule microarrays. AB - Current methods to identify interactions on small molecule microarrays (SMMs) introduce false positives that are difficult to dissect from the "real" binding events without tedious downstream re-evaluation. To specifically elucidate only activity-dependent ligand binding interactions, we have developed a technique that can be universally applied to present SMM systems. Our method makes use of a dual-color application strategy and is based on the simultaneous application of differentially treated samples. Overcoming the limitations of slide-to-slide variation, this method directly revealed activity-dependent interactions through a one-step application of protein samples on SMMs. Besides providing lead molecules for further development, the high-throughput screening results confer activity-dependent fingerprints for quantitative characterization and differentiation of proteins. The procedure was tested using a synthetic hydroxamate peptide library with 1400 discrete sequences permuted combinatorially across P1', P2', and P3' positions. Functional profiling across a panel of metalloproteases provided 44,800 datapoints within just eight SMM slides. These data were globally analyzed for activities, specificity, potency, and hierarchical clustering providing unique insights into inhibitor design and preference within this group of enzymes. Quantitative K(D) measurements performed on SMMs using one of the enzymes in the panel, Anthrax Lethal Factor, the toxic component of a notorious bioterror agent, unraveled several lead micromolar binders for further development. Overall, the effectiveness of the SMM platform is shown to be enhanced and extended using the strategy presented in this work. PMID- 17915868 TI - A chemical approach to the pharmaceutical optimization of an anti-HIV protein. AB - Chemical protein synthesis is important for dissecting the molecular basis of protein function. Here we advance its scope by demonstrating the significant improvement of the multifaceted pharmaceutical profile of small proteins exclusively via a chemical-based approach. The focus of this work centered on CCL 5 (RANTES) derivatives with potent anti-HIV activity. The overall chemical strategy involved a combination of coded and noncoded amino acid mutagenesis, peptide backbone engineering, and site-specific polymer attachment. The ability to alter specific protein residues, as well as precise control of the position and type of polymer attachment, allows for the exploration of specific molecular designs and resulted in novel CCL-5 analogues with significant differences in their respective biochemical and pharmaceutical properties. Using this approach, the complex-interplay of variables contributing to the noncovalent self association (aggregation) state, CCR-5 specificity, in vivo elimination half life, and anti-HIV activity of CCL-5-based protein analogues could be empirically evaluated via total chemical synthesis. This work has led to the identification of potent (sub-nanomolar) anti-HIV proteins with significantly improved pharmaceutical profiles, and illustrates the increasing value of protein chemical synthesis in contemporary therapeutic discovery. These antiviral molecules provide a novel mechanism of action for the development of a new generation of anti-HIV therapeutics which are still desperately needed. PMID- 17915869 TI - Formation of giant lipid vesiclelike compartments from a planar lipid membrane by a pulsed jet flow. PMID- 17915870 TI - Single molecule junctions formed via Au-thiol contact: stability and breakdown mechanism. AB - The stability and breakdown mechanism of a single molecule covalently bound to two Au electrodes via Au-S bonds were studied at room temperature. The distance over which a molecular junction can be stretched before breakdown was measured using a scanning tunneling microscopy break junction approach as a function of stretching rate. At low stretching rates, the stretching distance is small and independent of stretching rate. Above a certain stretching rate, it increases linearly with the logarithm of stretching rate. At very high stretching rates, the stretching distance reaches another plateau and becomes insensitive to the stretching rate again. The three regimes are well described by a thermodynamic bond-breaking model. A comparative study of Au-Au atomic point contacts indicates that the breakdown of the molecular junctions takes place at Au-Au bonds near the molecule-electrode contact. By fitting the experimental data with the model, the lifetime and binding energy were extracted. Both quantities are found to have broad distributions, owing to large variations in the molecule-electrode contact geometry. Although the molecular junctions are short-lived on average, certain contact geometries are considerably more stable. Several types of stochastic fluctuations were observed in the conductance of the molecule junctions, which are attributed to the atomic level rearrangement of the contact geometry, and bond breakdown and reformation processes. The possibility of bond reformation increases the apparent lifetime of the molecular junctions. PMID- 17915871 TI - Porphyrinoid chemistry in hemoprotein matrix: detection and reactivities of iron(IV)-oxo species of porphycene incorporated into horseradish peroxidase. PMID- 17915872 TI - At the water's edge: nitric acid as a weak acid. PMID- 17915873 TI - Self-construction of core-shell and hollow zeolite analcime icositetrahedra: a reversed crystal growth process via oriented aggregation of nanocrystallites and recrystallization from surface to core. AB - Zeolite analcime with a core-shell and hollow icositetrahedron architecture was prepared by a one-pot hydrothermal route in the presence of ethylamine and Raney Ni. Detailed investigations on samples at different preparation stages revealed that the growth of the complex single crystalline geometrical structure did not follow the classic crystal growth route, i.e., a crystal with a highly symmetric morphology (such as polyhedra) is normally developed by attachment of atoms or ions to a nucleus. A reversed crystal growth process through oriented aggregation of nanocrystallites and surface recrystallization was observed. The whole process can be described by the following four successive steps. (1) Primary analcime nanoplatelets undergo oriented aggregation to yield discus-shaped particles. (2) These disci further assemble into polycrystalline microspheres. (3) The relatively large platelets grow into nanorods by consuming the smaller ones, and meanwhile, the surface of the microspheres recrystallizes into a thin single crystalline icositetrahedral shell via Ostwald ripening. (4) Recrystallization continues from the surface to the core at the expense of the nanorods, and the thickness of the monocrystalline shell keeps on increasing until all the nanorods are consumed, leading to hollow single crystalline analcime icositetrahedra. The present work adds new useful information for the understanding of the principles of zeolite growth. PMID- 17915874 TI - Living radical polymerizations with germanium, tin, and phosphorus catalysts- reversible chain transfer catalyzed polymerizations (RTCPs). AB - A novel class of living radical polymerizations with germanium, tin, and phosphorus catalysts were developed. The polymerizations are based on a new reversible activation mechanism, Reversible chain Transfer (RT) catalysis. Low polydispersity (M(w)/M(n) approximately 1.1-1.3) polystyrene, poly(methyl methacrylate), poly(glycidyl methacrylate), and poly(2-hydroxyethyl methacrylate) with predicted molecular weight were obtained with fairly high conversion in a fairly short time. The pseudo-first-order activation rate constant kact for the styrene/GeI4 (catalyst) system was large enough, even with a small amount of GeI4, explaining why the system provides low-polydispersity polymers from an early stage of polymerization. The retardation in the polymerization rate observed for the styrene/GeI4 system was kinetically proven to be mainly due to the cross-termination between the propagating radical with GeI3*. Attractive features of the germanium, tin, and phosphorus catalysts include their high reactivity hence small amounts (1-10 mM) being required under relatively mild conditions (at 60-100 degrees C), high solubility in organic media without ligands, insensitivity to air hence sample preparation being allowed in the air, and minor color and smell. The germanium and phosphorus catalysts may also be attractive for their low toxicity. The phosphorus catalysts may also be attractive for their low cost. PMID- 17915875 TI - A new type of imido group donor: synthesis and characterization of sulfonylimino lambda(3)-bromane that acts as a nitrenoid in the aziridination of olefins at room temperature under metal-free conditions. PMID- 17915877 TI - Mechanism of alkyne insertion of a cationic zirconocene aryl complex. PMID- 17915876 TI - Engineering and analysis of a self-sufficient biosynthetic cytochrome P450 PikC fused to the RhFRED reductase domain. PMID- 17915878 TI - Deuterium exchange as an indicator of hydrogen bond donors and acceptors. PMID- 17915881 TI - Spiroiminodihydantoin as an oxo-atom transfer product of 8-oxo-2'-deoxyguanosine oxidation by chromium(V). AB - Oxidation of the DNA lesion 8-oxo-2'-deoxyguanosine by the two electron oxidants N,N'-ethylenebis(salicylideneanimato)oxochromium(V) (Cr(V)-salen) and bis(2-ethyl 2-hydroxybutyrato)oxochromium(V) (Cr(V)-ehba) at neutral pH forms spiroiminodihydantoin by an oxo-atom transfer mechanism. The chromium complexes are models of a DNA oxidation pathway caused by the carcinogen chromate. PMID- 17915882 TI - Synthesis of a neamine dimer targeting the dimerization initiation site of HIV-1 RNA. AB - A neamine dimer designed to bind to a specific sequence of HIV-1 RNA has been synthesized. Starting from neomycin B (1), a five-step synthesis efficiently provided a key protected neamine monomer 6 (28%). From the latter, coupling reactions with activated diacids gave dimers. After deprotection, a neamine dimer was obtained as the hexachlorohydrate salt 15 with 13% overall yield over nine steps. PMID- 17915883 TI - Binding affinities of oligonucleotides and PNAs containing phenoxazine and G clamp cytosine analogues are unusually sequence-dependent. AB - Melting temperatures of DNA duplexes containing the phenoxazine (P) and G-clamp (X) cytosine analogues exhibited a strong and unusual dependence on the nucleoside flanking the modified nucleobase, and the same trend was observed in PNA-DNA duplexes incorporating X in the PNA chain. Molecular dynamics simulations of the DNA duplexes show that generalized stacking (including secondary interactions of the ammonium group of X) and hydrogen bonding are good descriptors of the different duplex stabilities. PMID- 17915885 TI - Stereoselective synthesis of 2-deoxy-furanosides from 2,3-anhydro-furanosyl thioglycosides. AB - The stereocontrolled synthesis of 2-thiotolyl-furanosides from 2,3-anhydro furanosyl thioglycosides through a rearrangement-glycosylation process is reported. The efficiency of this reaction is high, providing 70-95% yields of the products. Treatment of the resulting 2-deoxy-2-thiotolyl-glycosides with hydrogen and Raney nickel affords the corresponding 2-deoxy-furanosides with a 1,3-syn relationship. PMID- 17915884 TI - Solid-phase synthesis of symmetrical 5',5'-dinucleoside mono-, di-, tri-, and tetraphosphodiesters. AB - Four classes of phosphitylating reagents were subjected to reactions with aminomethyl polystyrene resin-bound p-acetoxybenzyl alcohol to yield the corresponding polymer-bound mono-, di-, tri-, and tetraphosphitylating reagents. The solid-phase reagents were reacted with unprotected nucleosides (e.g., thymidine, adenosine, 3'-azido-3'-deoxythymidine, cytidine, or inosine) in the presence of 5-(ethylthio)-1H-tetrazole. Polymer-bound nucleosides underwent oxidation with tert-butyl hydroperoxide, deprotection of cyanoethoxy groups with DBU, and the acidic cleavage, respectively, to afford 5',5'-dinucleoside mono-, di-, tri-, and tetraphosphodiesters in 59-78% yield. PMID- 17915886 TI - A novel ambipolar spirobifluorene derivative that behaves as an efficient blue light emitter in organic light-emitting diodes. AB - A novel ambipolar spiro-configured D-A blue-light emitter bearing hole transporting diphenylamino groups and electron-transporting phenylbenzimidazole groups was synthesized, characterized, and incorporated into an efficient single layer organic light-emitting diode (OLED) device exhibiting blue-emission Commission International d'Eclairage (CIE) coordinates of 0.15 and 0.14, a turn on potential of 4 V, a maximum brightness of 2800 cd/m2 at 830 mA/cm2 (19 V), and a maximum quantum efficiency of 0.53% (0.61 cd/A). PMID- 17915887 TI - Aminocarbonylation of aryl halides using a nickel phosphite catalytic system. AB - The nickel and phosphite catalytic system with sodium methoxide enables a very efficient aminocarbonylation reaction to be performed between aryl iodides or bromides and N,N-dimethylformamide (DMF). Phosphite ligand 1, which is very stable to air and moisture and, furthermore, inexpensive, afforded the highest reaction yield. PMID- 17915888 TI - Synthesis of giant rigid pi-conjugated dendrimers. AB - A novel family of giant pi-conjugated dendrimers (G0, G1, and G2) solely constructed by 5,5,10,10,15,15-hexahexyltruxene units has been developed in a convergent manner through a Suzuki cross-coupling reaction. The overall yields to such large rigid conjugated dendrimers are quite satisfying. The structures and purity of these nanosize rigid dendrimers are verified by 1H and 13C NMR, MALDI TOF MS, and elemental analysis. PMID- 17915889 TI - Synthesis of fullerene glycoconjugates via a copper-catalyzed Huisgen cycloaddition reaction. AB - The synthesis of fullerene-carbohydrate conjugates using a copper-catalyzed [3+2] cycloaddition reaction to facilitate the union of an azido-functionalized sugar and a pentaalkynyl[60]fullerene is straightforward. Thus, fullerenes bearing five oligosaccharides such as Gb3-trisaccharide can be readily accessed. Nanometer scale molecular architectures presenting as many as 15 sugar moieties in C5 symmetry are readily produced. The cycloaddition reaction proceeds quantitatively under mild conditions without the need to protect the sugar hydroxyl groups. PMID- 17915891 TI - Solvent-controlled Csp2-->O silyl migration: the "one-pot" synthesis of 2,3 disubsituted thiophenes. AB - An effective "one-pot" synthesis of disubstituted thiophene derivatives employing 3-bromo-2-silyl thiophenes has been developed. A solvent-controlled [1,4] Csp2- >O silyl migration was involved as the key step in this process. PMID- 17915890 TI - Total synthesis of iejimalide B. An application of the Shiina macrolactonization. AB - The potent anticancer compound iejimalide B (1) was prepared by a total synthesis through a strategy that features Julia olefinations, Wittig olefinations, a Carreira enantioselective alkynylation, a Heck reaction, a Marshall propargylation reaction, a Stille coupling, and a Shiina macrolactonization. PMID- 17915893 TI - Determination of amine and aldehyde surface densities: application to the study of aged plasma treated polyethylene films. AB - The aim of this work was to test and to compare different methods reported in the literature to quantify amine and aldehyde functions on the surface of polyethylene (PE) films treated by ammonia plasma and to specify their stability against time. A low pressure ammonia plasma reactor was used to functionalize PE films with amine groups, which could be subsequently used for further immobilization of biomolecules. In order to determine the density of amine groups on the surface of treated films, various molecule probes and spectrophotometric analytical methods have been investigated. Two methods using (i) sulfosuccinimidyl 6-[3'-(2-pyridyldithio)-propionamido] hexanoate (sulfo-LC-SPDP) and (ii) 2-iminothiolane (ITL) associated with bicinchoninic acid (BCA) have been proved to be reliable and sensitive enough to estimate the surface concentration of primary amine functions. The amount of primary amino groups on the functionalized polyethylene films was found to be between 1.2 and 1.4 molecules/nm2. In a second step, the surface concentration of glutaraldehyde (GA), which is currently used as a spacer arm before immobilization of biomolecules, has been assessed: two methods were used to determine the surface density of available aldehyde functions, after the reaction of GA with the aminated polyethylene film. The concentration of GA was found to be in the same range as primary amine concentration. The influence of aging time on the density of available amino and aldehyde groups on the surfaces were evaluated under different storage conditions. The results showed that 50% of the initial density of primary amine functions remained available after storage during 6 days of the PE samples in PBS (pH 7.6) at 4 degrees C. In the case of aldehyde groups, the same percentage of the initial density (50%) remained active after storage in air at RT over a longer period, i.e., 15 days. PMID- 17915894 TI - Thermoresponsive micelles from Jeffamine-b-poly(L-glutamic acid) double hydrophilic block copolymers. AB - Double hydrophilic block copolymers (DHBC) consisting of a Jeffamine block, a statistical copolymer based on ethylene oxide and propylene oxide units possessing a lower critical solution temperature (LCST) of 30 degrees C in water, and poly(L-glutamic acid) as a pH-responsive block were synthesized by ring opening polymerization of gamma-benzyl-L-glutamate N-carboxyanhydride using an amino-terminated Jeffamine macroinitiator, followed by hydrolysis. This DHBC proved thermoresponsive as evidenced by dynamic light scattering and small-angle neutron scattering experiments. Spherical micelles with a Jeffamine core and a poly(L-glutamic acid) corona were formed above the LCST of Jeffamine. The size of the core of such micelles decreased with increasing temperature, with complete core dehydration being achieved at 66 degrees C. Such behavior, commonly observed for thermosensitive homopolymers forming mesoglobules, is thus demonstrated here for a DHBC that self-assembles to generate thermoresponsive micelles of high colloidal stability. PMID- 17915895 TI - Rapid evaluation and screening of interfacial reactions on self-assembled monolayers. AB - This paper reports 16 chemical reactions for elaborating the structures of self assembled monolayers (SAMs) of alkanethiolates on gold. This work takes advantage of matrix-assisted laser desorption/ionization and time-of-flight mass spectrometry (MALDI-TOF MS) to rapidly characterize the products and yields of reactions that occur with molecules attached to monolayers. The paper also describes a method for screening reaction conditions, wherein monolayers are treated with an array of reactants and mass spectrometry is used to identify those regions that undergo reactions to give new, and unanticipated, products in high yield. These examples serve to increase the collection of reactions that can be used to elaborate the structures, and therefore the properties, of self assembled monolayers of alkanethiolates on gold and to introduce label-free methods for screening interfacial reactions. PMID- 17915896 TI - Cell and protein compatibility of parylene-C surfaces. AB - Parylene-C, which is traditionally used to coat implantable devices, has emerged as a promising material to generate miniaturized devices due to its unique mechanical properties and inertness. In this paper we compared the surface properties and cell and protein compatibility of parylene-C relative to other commonly used BioMEMS materials. We evaluated the surface hydrophobicity and roughness of parylene-C and compared these results to those of tissue culture treated polystyrene, poly(dimethylsiloxane) (PDMS), and glass. We also treated parylene-C and PDMS with air plasma, and coated the surfaces with fibronectin to demonstrate that biochemical treatments modify the surface properties of parylene C. Although plasma treatment caused both parylene-C and PDMS to become hydrophilic, only parylene-C substrates retained their hydrophilic properties over time. Furthermore, parylene-C substrates display a higher degree of nanoscale surface roughness (>20 nm) than the other substrates. We also examined the level of BSA and IgG protein adsorption on various surfaces and found that surface plasma treatment decreased the degree of protein adsorption on both PDMS and parylene-C substrates. After testing the degree of cell adhesion and spreading of two mammalian cell types, NIH-3T3 fibroblasts and AML-12 hepatocytes, we found that the adhesion of both cell types to surface-treated parylene-C variants were comparable to standard tissue culture substrates, such as polystyrene. Overall, these results indicate that parylene-C, along with its surface-treated variants, could potentially be a useful material for fabricating cell-based microdevices. PMID- 17915897 TI - Membrane curvature effects on glycolipid transfer protein activity. AB - The glycolipid transfer protein (GLTP) is monomeric in aqueous solutions, and it binds weakly to membrane interfaces with or without glycolipids. GLTP is a surface-active protein and adsorbs to exert a maximal surface pressure value of 19 mN/m. The change in surface pressure following GLTP adsorption decreased linearly with initial surface pressure. The exclusion pressure for different phospholipids and sphingolipids was between 23 and 31 mN/m, being clearly highest for the negatively charged dipalmitoyl-phosphatidylserine. This can be explained by electrostatic forces when GLTP is positively charged at neutral pH (isoelectric point = 9.0) and by phosphatidylserine being negatively charged. If GLTP is injected under a palmitoyl-galactosylceramide monolayer above 30 mN/m, the presence of GLTP leads to a decrease in the surface pressure as a function of time. This suggests that GLTP is able to remove glycolipids from the monolayer without penetrating the monolayer. On the other hand, if phospholipid vesicles with or without glycolipids are also present in the subphase, no change in the surface pressure takes place. This suggests that GLTP in the presence of curved membranes is not able to transfer from or to planar membranes. We also show that transfer of fluorescently labeled galactosylceramide is faster from small highly curved palmitoyl-oleoyl-phosphatidylcholine and dipalmitoyl-phosphatidylcholine bilayer vesicles but not from palmitoyl-sphingomyelin vesicles regardless of the size. PMID- 17915898 TI - Structural characterization of frozen n-heptane solutions of metal-containing reverse micelles. AB - The microstructure of temperature-quenched solutions of reverse micelles formed by sodium, cobalt, ytterbium, and cobalt/ytterbium bis(2 ethylhexyl)sulfosuccinate in n-heptane has been investigated by SAXS and EXAFS. Some changes in the X-ray absorption spectra with respect to the same systems at room temperature have been observed. The analysis of the SAXS spectra leads to the hypothesis that at 77 K the closed spherical structure of reverse micelles is retained and that during the temperature quench they undergo a clustering process involving the transition from a quite random dispersion to the formation of more or less large clusters of strongly packed reverse micelles. This behavior is attributed to competitive effects caused by the temperature decrease. The prevalence of intermicellar attractive interactions with respect to Brownian motions leading to a collapse to more compact structure is in competition with the rapid decrease of reverse micelle diffusion rate involving a freezing of the local structures. In the case of cobalt, ytterbium, and cobalt/ytterbium bis(2 ethylhexyl)sulfosuccinate reverse micelles, further information from EXAFS measurements indicates that within the reverse micelle core exists a quite ordered nanosized domain composed of water, surfactant counterions, and oxygen atoms of the SO3- head groups. The conservation of local order and inverse structure during the clustering phenomenon that results from the fast freezing with liquid nitrogen of solutions of reverse micelles could have biological implications, i.e., the preservation of tissue samples at cryogenic temperatures. PMID- 17915899 TI - Combining hydrogen-bonding complexation in solution and hydrogen-bonding-directed layer-by-layer assembly for the controlled loading of a small organic molecule into multilayer films. AB - We have combined hydrogen-bonding complexation in solution and layer-by-layer assembly for the controlled loading of a water-insoluble small organic molecule, bis-triazine (DTA), an azobenzene derivative containing multiple hydrogen bond donors and acceptors, into layer-by-layer multilayer films of poly(acrylic acid) and diazo-resin. UV-visible spectroscopy indicates that DTA has been loaded into multilayer films, with the loading amount increasing linearly with the number of layers. The loading amount can be well tuned either by changing the concentration of DTA or the solvent composition at the complexation step. Fourier transform infrared spectroscopy has revealed that both the complexation and layer-by-layer assembly are driven by hydrogen bonding. After photo-cross-linking and immersion in dimethyl sulfoxide to release DTA, the film can serve as an absorbent for DTA. This study provides a new unconventional layer-by-layer assembly that combines hydrogen-bonding complexation in solution and hydrogen-bond-driven layer-by-layer assembly at the interface. This method provides a new route to load a variety of water-insoluble functional organic molecules into layer-by-layer films. PMID- 17915900 TI - One-step synthesis of highly dispersed gold nanocrystals on silica spheres. AB - Highly dispersed gold nanocrystals decorating silica spheres were prepared from HAuCl4 and NaOH via a deposition-precipitation (DP) process, in which the isoelectric point (IEP) of the substrate was adjusted during sphere synthesis by interaction of the surface with ammonia molecules. Through the systematic variation of pH (4-8), reaction temperature (65-96 degrees C), and time (10-30 min), a superior product with small (2-5 nm), homogeneously distributed gold crystals was obtained at pH 7 and a reaction temperature of 96 degrees C. These materials will offer enhanced performance as catalysts and contrast enhancers in biomedical imaging. PMID- 17915901 TI - Effect of particle size on the kinetics of the electrocatalytic oxygen reduction reaction catalyzed by Pt dendrimer-encapsulated nanoparticles. AB - Platinum dendrimer-encapsulated nanoparticles (DENs) containing an average of 55, 100, 147, 200, and 240 atoms were prepared within sixth-generation, hydroxyl terminated, poly(amidoamine) dendrimers. These DENs were immobilized on glassy carbon electrodes, and the effect of particle size on the kinetics of the oxygen reduction reaction (ORR) was quantitatively evaluated using rotating disk voltammetry. The total areas of the Pt DENs were determined by electrochemical CO stripping and hydrogen desorption, and the results were found to be in reasonable agreement with calculated values. The largest particles exhibited the highest specific activities for the ORR. PMID- 17915902 TI - Gold nanoparticle-modified ITO electrode for electrogenerated chemiluminescence: well-preserved transparency and highly enhanced activity. AB - Although it is desirable to use transparent indium tin oxide (ITO)-coated glass substrates as working electrodes for electrogenerated chemiluminescence (ECL), their applications in ECL studies of the Ru(bpy)32+ (bpy, 2,2'-bipyridine)/tri-n propylamine (TPrA) system have been limited because of the large overpotential of TPrA oxidation and the instability of the ITO surface at high anodic potentials. Here, we describe a simple method to achieve high ECL activity using ITO electrodes modified with gold nanoparticles (GNPs). The GNPs have been capped with fluorosurfactant ligands (i.e., Zonyl FSO). Much more facile TPrA oxidation was achieved by using the modified electrode, and an intense low-oxidation potential (LOP) ECL signal was observed at approximately 0.88 V versus SCE. The electrode transmittance drop upon modification was generally less than 5% over the visible spectrum when small-sized GNPs (approximately 4 nm) were employed. The well-preserved transparency and highly enhanced activity make the modified electrode promising for ECL studies. PMID- 17915903 TI - Electrochemical synthesis and surface characterization of poly(3,4 ethylenedioxythiophene) films grown in an ionic liquid. AB - We report a facile method to synthesize poly(3,4-ethylenedioxythiophene) (PEDOT) films at room temperature in a waterproof ionic liquid, 1-ethyl-3 methylimidazolium bis(perfluoroethylsulfonyl)imide (EMIPFSI), by electropolymerization. The ionic liquid leads to the formation of randomly oriented nanofibers and particles confined to submicrometer-sized domains in the film microstructure. X-ray photoelectron spectroscopy (XPS) and energy-dispersive X-ray (EDX) studies provide information about the intercalation of the cation apart from the reported anion in the polymer film, and on how the imidazolium ion controls the growth of PEDOT nanostructures. PMID- 17915904 TI - Adsorption characteristics of thionine on gold nanoparticles. PMID- 17915905 TI - Prediction and classification of drug toxicity using probabilistic modeling of temporal metabolic data: the consortium on metabonomic toxicology screening approach. AB - Detection and classification of in vivo drug toxicity is an expensive and time consuming process. Metabolic profiling is becoming a key enabling tool in this area as it provides a unique perspective on the characterization and mechanisms of response to toxic insult. As part of the Consortium on Metabonomic Toxicology (COMET) project, a substantial metabolic and pathological database was constructed. We chose a set of 80 treatments to build a modeling system for toxicity prediction using NMR spectroscopy of urine samples (n=12935) from laboratory rats (n=1652). The compound structures and activities were diverse but there was an emphasis on the selection of hepato and nephrotoxins. We developed a two-stage strategy based on the assumptions that (a) adverse effects would produce metabolic profiles deviating from those of normal animals and (b) such deviations would be similar for treatments having similar physiological effects. To address the first stage, we developed a multivariate model of normal urine, using principal components analysis of specially preprocessed 1H NMR spectra. The model demonstrated a high correspondence between the occurrence of toxicity and abnormal metabolic profiles. In the second stage, we extended a density estimation method, "CLOUDS", to compute multidimensional similarities between treatments. Crucially, the technique allowed a distribution-free estimate of similarity across multiple animals and time points for each treatment and the resulting matrix of similarities showed segregation between liver toxins and other treatments. Using the similarity matrix, we were able to correctly identify the target organ of two "blind" treatments, even at sub-toxic levels. To further validate the approach, we then applied a leave-one-out approach to predict the main organ of toxicity (liver or kidney) showing significant responses using the three most similar matches in the matrix. Where predictions could be made, there was an error rate of 8%. The sensitivities to liver and kidney toxicity were 67 and 41%, respectively, whereas the corresponding specificities were 77 and 100%. In some cases, it was not possible to make predictions because of interference by drug-related metabolite signals (18%), an inconsistent histopathological or urinary response (11%), genuine class overlap (8%), or lack of similarity to any other treatment (2%). This study constitutes the largest validation to date of the metabonomic approach to preclinical toxicology assessment, confirming that the methodology offers practical utility for rapid in vivo drug toxicity screening. PMID- 17915906 TI - "Turnover proteome" of human atrial trabeculae. AB - Most of the biologically relevant data on cardiomyocytes are derived from isolated cells under conditions that are, to some extent, altered compared to the natural milieu of the functional heart. The handling procedure of the dissection, isolation, and short-term culturing induces changes in the cells such that the subsequently measured parameters (among others, the protein synthesis) reflect the actual experimental conduct rather than the intrinsic properties of these terminally differentiated cells. Although it is known that the protein synthetic machinery of isolated cardiomyocytes is operational and functional, the biosynthetic yield of human cardiomyocytes in the natural milieu of the trabeculae remains to be established, with a special emphasis to clarify whether the protein synthesis includes just a limited set of polypeptides or it encompasses all cellular constituents. Knowledge on this issue is a prerequisite for achieving further advances in our understanding of heart remodeling related to hypertrophy in particular, and for attempting interventions leading to repair of damaged heart in general. The experimental system of "organ bath" enables simultaneous registration of contractile forces of portions of cardiac muscle tissue (and other myocyte-containing tissues) and biosynthetic labeling of newly synthesized cellular constituents. The organ bath methodology was adapted for this project such as enabling to measure molecular changes in response to in vitro applied stimuli. Instead of Krebs-Henseleit-solution, as used in classical protocols of organ bath studies, we utilized cell culture media suitable to experimental conditions related to metabolic labeling. Proteome patterns established by performing two-dimensional gel electrophoresis of the extracts from biosynthetically labeled trabeculae revealed that cardiomyocytes synthesize the full spectrum of cellular proteins. Proteomic silver-stain readout was used to obtain samples for spot excisions, as material suitable for mass spectrometric analysis. Protein spots were identified both from the excised spots and also by matching with the in-house- and www-databases (Swiss-Prot/High-Performance Heart). From our findings that protein synthesis in terminally differentiated cardiomyocytes is not confined just to the synthesis of those structures needed for the post-mitotic house-keeping functions, we conclude that this model might serve as a valid experimental system to study and elucidate the effects of various pharmacological compounds under conditions where physiology (contractile forces) and biochemistry (protein synthesis) of intact human heart tissue are monitored simultaneously. PMID- 17915908 TI - Proteomic approach for caudal trauma-induced acute phase proteins reveals that creatine kinase is a key acute phase protein in amphioxus humoral fluid. AB - Elevated creatine kinase (CK) in the circulation was generally regarded to be a passive release from muscle damage. We utilized proteomic methodologies to characterize amphioxus humoral fluid APPs in response to caudal trauma, and found several spots of CK alterations with up-regulation and pI shift. Its amount and enzyme activity showed a dynamic pattern of APP in humoral fluid accompanied with a reduction in enzyme activity of muscle, whereas there was no significant difference in CK amount of muscle and the other tissues and in CK enzyme activity of the other tissues between different time points of sample collection following caudal trauma. In addition, CK phosphorylation regulation during injury was not achieved by monoclonal antibodies separately against phosphothreonine, phosphotyrosine, and phosphoserine. These results suggested that the CK elevation of humoral fluid might be from muscle, being an active response to caudal trauma rather than a passive release from muscle damage. Therefore, CK ability in response to caudal trauma should be highly concerned. PMID- 17915909 TI - Chromophore localization in conjugated polymers: molecular dynamics simulation. AB - Quantum chemical calculations of undistorted poly(phenylene vinylene) chains at zero temperature exhibit chromophores which are delocalized over the whole polymer. We demonstrate with molecular dynamics simulations that chromophore localization in agreement with experiment can be obtained if the system is simulated at finite temperature. The dependence of the chromophore localization on the temperature is investigated. PMID- 17915907 TI - Glycomics of proteoglycan biosynthesis in murine embryonic stem cell differentiation. AB - Glycosaminoglycans (GAGs) play a critical role in binding and activation of growth factors involved in cell signaling critical for developmental biology. The biosynthetic pathways for GAGs have been elucidated over the past decade and now analytical methodology makes it possible to determine GAG composition in as few as 10 million cells. A glycomics approach was used to examine GAG content, composition, and the level of transcripts encoding for GAG biosynthetic enzymes as murine embryonic stem cells (mESCs) differentiate to embryoid bodies (EBs) and to extraembryonic endodermal cells (ExE) to better understand the role of GAGs in stem cell differentiation. Hyaluronan synthesis was enhanced by 13- and 24-fold, most likely due to increased expression of hyaluronan synthase-2. Chondroitin sulfate (CS)/dermatan sulfate (DS) synthesis was enhanced by 4- and 6-fold, and heparan sulfate (HS) synthesis was enhanced by 5- and 8-fold following the transition from mESC to EB and ExE. Transcripts associated with the synthesis of the early precursors were largely unaltered, suggesting other factors account for enhanced GAG synthesis. The composition of both CS/DS and HS also changed upon differentiation. Interestingly, CS type E and highly sulfated HS both increase as mESCs differentiate to EBs and ExE. Differentiation was also accompanied by enhanced 2-sulfation in both CS/DS and HS families. Transcript levels for core proteins generally showed increases or remained constant upon mESC differentiation. Finally, transcripts encoding selected enzymes and isoforms, including GlcNAc-4,6-O-sulfotransferase, C5-epimerases, and 3-O-sulfotransferases involved in late GAG biosynthesis, were also enriched. These biosynthetic enzymes are particularly important in introducing GAG fine structure, essential for intercellular communication, cell adhesion, and outside-in signaling. Knowing the changes in GAG fine structure should improve our understanding the biological properties of differentiated stem cells. PMID- 17915910 TI - Molecular orientation of membrane-anchored mucin glycoprotein mimics. AB - Mucin glycoproteins contribute to a wide range of cell-surface phenomena. Their dense glycosylation is believed to confer structural rigidity as well as molecular extension beyond the glycocalyx, crucial to interaction with the cellular environment. However, controlled investigations of the relationships between glycosylation, rigidity, and extension of membrane-bound mucins or similar macromolecules are lacking, largely because of the absence of tractable experimental models. We have therefore made use of recently developed synthetic mucin mimetics, in which the core alpha-GalNAc monosaccharides of natural mucins are conjugated to a lipidated polymer backbone and anchored to fluid, solid supported lipid membranes, and fluorescence interference contrast microscopy, an optical technique that provides nanometer-scale topographic information about objects near a reflective interface, to measure the orientation of the mucin mimics relative to the membrane plane. Data from two independent probes, fluorophores conjugated directly to the polymer backbone and fluorescent proteins bound to the sugar groups, unexpectedly show that the mucin mimic molecules lie flat along the membrane. Rigidity and core glycosylation are therefore insufficient to ensure molecular projection outward from a membrane surface. PMID- 17915911 TI - Molecular dynamics study of the solvation of calcium carbonate in water. AB - We performed molecular dynamics simulations of diluted solutions of calcium carbonate in water. To this end, we combined and tested previous polarizable models. The carbonate anion forms long-living hydrogen bonds with water and shows an amphiphilic character, in which the water molecules are expelled in a region close to its C(3) symmetry axis. The calcium cation forms a strongly bound ion pair with the carbonate. The first hydration shell around the CaCO(3) pair is found to be very similar to the location of the water molecules surrounding CaCO(3) in ikaite, the hydrated mineral. PMID- 17915912 TI - Oil-induced anomalous thermoresponsive viscoelasticity in fluorinated surfactant systems. AB - We have studied the rheology and structure of a mixed nonionic fluorinated surfactant, perfluoroalkyl sulfonamide ethoxylate, C(8)F(17)SO(2)N(C(3)H(7))(CH(2)CH(2)O)(n)H abbreviated as C(8)F(17)EO(10), and perfluorodecalin (C(10)F(18)) or perfluoropolyether oil, (C(3)F(6)O)(n)COOH, in an aqueous system using rheometry and small-angle X-ray scattering (SAXS) techniques. In the absence of oil, the viscosity of surfactant solutions (10 and 15 wt %) first decreases slightly and then more strongly with temperature. Addition of a small amount of fluorinated oil to the wormlike micellar solution disrupts the network structure and decreases the viscosity sharply at lower temperature indicating a rod-sphere transition. The trend of the viscosity curve changes gradually and an anomalous viscosity maximum as a function of temperature appears. It is found that perfluoropolyether oil decreases the viscosity more effectively than perfluorodecalin. The generalized indirect Fourier transformation (GIFT) analysis of the SAXS data confirmed the formation of long rod-like particles in an oil-free, surfactant/water system at 20 degrees C. Addition of a trace amount of fluorinated oils induces modulation in the structure of the micelles and eventually short rods or spherical particles are formed. The decreasing trend in the viscosity with oil concentration is thus attributed to the microstructure changes induced by the added oils. PMID- 17915913 TI - NMR provides checklist of generic properties for atomic-scale models of periodic mesoporous silicas. AB - MCM-41 and SBA-15 silicas were studied by (29)Si solid-state NMR and (15)N NMR in the presence of (15)N-pyridine with the aim to formulate generic structural parameters that may be used as a checklist for atomic-scale structural models of this class of ordered mesoporous materials. High-quality MCM-41 silica constitutes quasi-ideal arrays of uniform-size pores with thin pore walls, while SBA-15 silica has thicker pore walls with framework and surface defects. The numbers of silanol (Q(3)) and silicate (Q(4)) groups were found to be in the ratio of about 1:3 for MCM-41 and about 1:4 for our SBA-15 materials. Combined with the earlier finding that the density of surface silanol groups is about three per nm(2) in MCM-41 (Shenderovich, et al. J. Phys. Chem. B 2003, 107, 11924) this allows us to discriminate between different atomic-scale models of these materials. Neither tridymite nor edingtonite meet both of these requirements. On the basis of the hexagonal pore shape model, the experimental Q(3):Q(4) ratio yields a wall thickness of about 0.95 nm for MCM-41 silica, corresponding to the width of ca. four silica tetrahedra. The arrangement of Q(3) groups at the silica surfaces was analyzed using postsynthesis surface functionalization. It was found that the number of covalent bonds to the surface formed by the functional reagents is affected by the surface morphology. It is concluded that for high-quality MCM-41 silicas the distance between neighboring surface silanol groups is greater than 0.5 nm. As a result, di- and tripodical reagents like (CH(3))(2)Si(OH)(2) and CH(3)Si(OH)(3) can form only one covalent bond to the surface. The residual hydroxyl groups of surface-bonded functional reagents either remain free or interact with other reagent molecules. Accordingly, the number of surface silanol groups at a given MCM-41 or SBA-15 silica may not decrease but increase after treatment with CH(3)Si(OH)(3) reagent. On the other hand, nearly all surface silanol groups could be functionalized when HN(Si(CH(3))(3))(2) was used. PMID- 17915914 TI - Determination of the parameters controlling swelling of chemically cross-linked pH-sensitive poly(N-vinylimidazole) hydrogels. AB - The number of variables controlling the behavior of ionic gels is large and very often some of them are unknown. The aim of this work is to interpret quantitatively the swelling behavior of pH sensitive gels, with the minimum number of simplifying assumptions. With this purpose, the equilibrium degree of swelling (S) and protonation (alpha) of chemically cross-linked poly(N vinylimidazole) (PVI) immersed in aqueous salt solutions were measured as a function of the ionic strength (mu), in the whole range of pH. In acid solutions with pH in the range 0 to 4, imidazole moieties become protonated, and PVI behaves as a polyelectrolyte gel: S decreases upon increasing mu both for NaCl and for CaCl(2), with HCl as protonating acid. In aqueous solutions with larger pH, between 4 and 12, the hydrogel is practically neutral, and S increases as mu rises, showing a salting-in effect. From the quantitative analysis of these results, the following facts emerged. Protonation induces chain stiffness (as measured by the non-Gaussian factor) and worsening of the solvent quality of the aqueous media (as measured by the polymer-solvent interaction parameter). For alpha below 33%, swelling seems to be governed by the excess of mobile counterions inside the gel with respect to the bath, with a minor but still significantly negative contribution of the osmotic swelling pressure due to polymer-solvent mixing. Above 33% protonation, it is necessary to consider Manning counterion condensation to get parameters with physical meaning. The crossover between polyelectrolyte and salting-in effects corresponds to alpha and mu values with the same ionic and mixing contributions to the osmotic swelling pressure. The formation of ionic nonpermanent cross-links, with H(2)SO(4) as the protonating acid, was discarded. PMID- 17915916 TI - An electrochemical gate based on a stimuli-responsive membrane associated with an electrode surface. AB - The electrochemical gate based on a chemical signal-responsive membrane was assembled on a Au electrode surface. The polyelectrolyte gel membrane was capable to bind cholesterol because of the hydrogen bonding between cholesterol and the polymer backbone resulting in the gel swelling. The membrane channels were reversibly closed and opened upon addition and washing out cholesterol, respectively. Thus, the electrochemical process of a soluble redox probe, [Fe(CN)(6)](3-/4-), at the membrane-modified electrode was reversibly switched "on-off" by the cyclic addition and washing out cholesterol. The electrochemical reaction was also tuned by the variation of the concentration of the added cholesterol that controlled the extent of the channels closing. The switchable and tuneable operation of the chemically controlled electrochemical gate was characterized by Faradaic impedance spectroscopy and atomic force microscopy, indicating that the extent of the pores opening and closing is controlled by the concentration of the membrane-associated cholesterol. The chemical-responsive electrochemical gate was suggested to be a part of future biochemical/electrochemical systems with logic operations. PMID- 17915917 TI - Effect of orientation on the physical properties of potato amylose and high amylose corn starch films. AB - The effect of orientation on the properties of amylose and starch films was studied in order to determine if film strength, flexibility, and water resistance could be improved. Potato amylose and high (70%) amylose corn starch were peracetylated, cast into films, stretched in hot glycerol 1-6 times the original length, and deacetylated. Molecular orientation of potato amylose films was much higher than for high-amylose corn starch films as determined by optical birefringence. For potato amylose films, orientation resulted in large increases in tensile strength and elongation but little change in modulus. For high-amylose corn starch films, tensile strength and modulus did not change with draw ratio but elongation to break increased from about 8% to 27% as draw ratio increased from 1 to 5. Scanning electron micrographs revealed many small crazes in the drawn starch films, suggesting that the improved film toughness was due to energy dissipation during deformation of the crazes. Annealing of drawn films at 100% humidity resulted in partial crystallization and improved wet strength. PMID- 17915918 TI - Smaller building blocks form larger assemblies: aggregation behavior of biaryl based dendritic facial amphiphiles. AB - Synthesis and micellar behavior of biaryl-based benzyl ether dendritic molecules prepared from a new biaryl building block are described. The key objective of the study is to tune the size of individual dendritic molecules and investigate its effect on aggregation behavior of the resulting micelle-like assemblies. We show that the functional group placement in the building block influences flexibility of the dendritic backbone and interior volume available for packing the hydrophobic groups, which is reflected in different aggregation behavior and aggregate size of the two types of micellar assemblies. PMID- 17915920 TI - Sequential electrochemical oxidation of alternating benzene-furan oligomers. AB - Electrochemical oxidation of pentaaryl 2 containing two furan moieties occurs sequentially to give diketone 8 after two-electron transfer. Further oxidation with another two-electron transfer gives the corresponding tetraketone 9. Radical cation intermediate is detected by absorption spectroscopy. The radical intermediates of different regiochemistry have been shown to exhibit different oxidation potentials as revealed by the differential pulse voltammetry. PMID- 17915919 TI - Asymmetric aerobic oxidation of alpha-hydroxy acid derivatives by C4-symmetric, vanadate-centered, tetrakisvanadyl(V) clusters derived from N-salicylidene-alpha aminocarboxylates. AB - A series of chiral vanadyl(V) methoxides bearing 3-t-butyl-5-substituted N salicylene-L-valinate and L-t-leucinate as chiral auxiliaries has been prepared. In all cases except the 3,5-di-t-butyl analogue, they exist as monomers both in solution and in the single crystal state. In the case of the 3,5-di-t-butyl analogue, the architectural nature of the vanadyl(V) complex highly depends on the base used during the complex formation event. A pentanuclear C4-symmetric complex was formed when potassium salts were employed instead of the corresponding sodium salts. A central vanadate(V) unit serves to grip four identical chiral monomeric vanadyl(V) units together, by which a potassium ion sits on top of the four flanking units through carbonyl coordinations and serves to hold the whole cluster by cooperation with the central vanadate(V) unit. In comparison with the corresponding monomeric vanadyl(V) methoxide complex, the cluster complex was utilized to facilitate the asymmetric aerobic oxidations of various racemic alpha-hydroxyesters, -amides, and -thioesters with excellent selectivity factors (krel 40 to >500). PMID- 17915921 TI - All-cis helical polypeptides. AB - The possibility of all-cis open-chain polypeptides is rarely addressed, owing to three main reasons, namely, (i) the extreme scarcity of cis peptide bonds in naturally occurring proteins and peptides, (ii) the lesser thermodynamic stability (by about 2.5 kcal/mol) of cis amide bonds with respect to their trans counterparts, and (iii) widely held preconceptions about the so-called "steric clash" between lateral chains borne by two successive alpha carbons. Quantum chemistry calculations performed on alanine tridecamers show how the latter constraints can be efficiently relieved through proper phi/psi adjustments along the backbone, leading to several helical arrangements--presumably the only permitted regular structures. Four more-or-less regular helices were thus characterized, one of them, a superhelix, exhibiting intramolecular hydrogen bonds. Understanding and anticipating all-cis open-chain structures not only make use of the classical Ramachandran maps at each C alpha i, relating to E = f(phi i,psi i), but also require the profile of a new kind of conformational dependence, the plaque maps, relating to E = f(phi i,psi i-1). The obvious coupling between two such maps enforces conformational dependence between two consecutive C alpha's, somewhat questioning in this context the customary "local effects", and presumably reducing the whole chain plasticity. Whereas cis thermodynamic penalty cannot be abolished locally, energy clues indicate that assembling cis-prepared building units is an exothermic process. Besides, once built up, the all-cis backbone should be difficult to unlock, thus affording reasonable kinetic stability. PMID- 17915922 TI - Interaction of C-Si, C-Sn, and Si-Si sigma-bonds with chalcogen lone pairs. AB - The ability of neighboring C-Si, C-Sn, and Si-Si groups in conformationally constrained cyclic molecules to reduce the lowest ionization energies of sulfur, selenium, and tellurium compounds has been determined by charge-transfer spectroscopy of complexes with tetracyanoethylene. For selected compounds, ionization energies were determined by gas-phase photoelectron spectroscopy. The lowest ionization energies measured by photoelectron spectroscopy, with one exception, correlate with the charge-transfer spectroscopic data. In addition, theoretical analysis has provided insight into the photoelectron spectra and the geometry-dependent interaction between C-Si or C-Sn bonds and chalcogen lone pairs. Substantial lowering of ionization energies is found which is anticipated to have important consequences in the chemistry of these and related species. PMID- 17915923 TI - Synthesis and reactivity of 1-substituted 2-fluoro- and 2,2-difluoroaziridines. AB - A straightforward synthesis toward 2-fluorinated aziridines was developed via ring closure of beta-fluorinated beta-chloroamines, which were obtained via reduction of the corresponding alpha-fluorinated amides by borane. When 1-benzyl 2-fluoroaziridine was treated with methanol, reaction occurred at the 2-position, giving rise to N-benzyl-2,2-dimethoxyethylamine, while in the case of 1-benzyl 2,2-difluoroaziridine the 3-position was attacked, giving rise to N-benzyl-2 methoxyacetamide. These reactions point to the divergent reactive behavior of monofluoro- and difluoroaziridines. PMID- 17915924 TI - A new class of discotic mesogens derived from tris(N-salicylideneaniline)s existing in C3h and Cs keto-enamine forms. AB - Two series of a unique class of columnar liquid crystals derived from tris(N salicylideneaniline)s [TSANs] in which the proton and the electron interact with each other through the H-bonding environment are reported. The synthesis is carried out by condensing 1,3,5-triformylphloroglucinol with the respective dialkoxyanilines or trialkoxyanilines. 1H NMR and 1H-1H COSY NMR studies revealed their existence as an inseparable mixture of two keto-enamine tautomeric forms with C3h and Cs rotational symmetries instead of the expected enol-imine form. The influence of the number of peripheral alkoxy tails on the columnar mesomorphic behavior is investigated by using polarizing optical microscopy, differential scanning calorimetry, and X-ray scattering. The fluid/glassy columnar states probed for a number of representative compounds confirmed the D6h (hexagonal) or D2h (rectangular) symmetry of the columns. The electronic absorption and emission characteristics of these compounds have been studied in both mesomorphic and solution states. Of special interest, the photoluminescence spectra of solution and fluid/glassy two-dimensional structure evidently disclose the promising light generating capability of these new discotics systems. PMID- 17915925 TI - Oligo(2,7-fluorene ethynylene)s with pyrene moieties: synthesis, characterization, photoluminescence, and electroluminescence. AB - A series of highly fluorescent, pyrene-modified oligo(2,7-fluorene ethynylene)s were synthesized and fully characterized. Different emissions were observed between pyrene-end-capped and pyrene-centered oligo(2,7-fluorene ethynylene)s. Moreover, these oligomers were applied to fabricate organic light-emitting diodes (OLEDs), and bright blue or green electroluminescence (EL) with good device performances was achieved in multilayer OLEDs. These pyrene-modified oligo(2,7 fluorene ethynylene)s could be used as optoelectronics materials or models to investigate fluorescent structure-property relationship of fluorene derivatives. PMID- 17915926 TI - Nucleofugality of phenyl and methyl carbonates. AB - A series of X,Y-substituted benzhydryl phenyl carbonates 1 and X,Y-substituted benzhydryl methyl carbonates 2 were subjected to solvolysis in different methanol/water, ethanol/water, and acetone/water mixtures at 25 degrees C. The LFER equation, log k = sf(Ef + Nf), was used to derive the nucleofuge-specific parameters (Nf and sf) for phenyl carbonate (1LG) and methyl carbonate (2LG) leaving groups in a given solvent in SN1 type reaction. Kinetic measurements showed that phenyl carbonates solvolyze one order of magnitude faster than methyl carbonates. Optimized geometries of 1LG and 2LG at B3LYP/6-311G(d,p), B3LYP/6 311++G(d,p), and MP2(full)/6-311++G(d,p) levels revealed that negative charge delocalization in carbonate anions to all three oxygen atoms occurs due to negative hyperconjugation. Phenyl carbonate (1LG) is a better leaving group (Nf = -0.84 +/- 0.07 in 80% v/v aq EtOH) than methyl carbonate 2LG (Nf = -1.84 +/- 0.07 in 80% v/v aq EtOH) because of more pronounced negative hyperconjugation, which is characterized with a more elongated RO-C bond and more increased RO-C-CO angle in 1LG than in 2LG. Calculated affinities of benzhydryl cation toward methyl and phenyl carbonate anions (DeltaDeltaEaff = 11.7 kcal/mol at the B3LYP/6 311++G(d,p) level and DeltaDeltaEaff = 2.7 kcal/mol at the PCM-B3LYP/6 311++G(d,p) level in methanol, respectively) showed that 1LG is more stabilized than 2LG, which is in accordance with greater solvolytic reactivity of 1 than 2. PMID- 17915927 TI - New cruciform structures: toward coordination induced single molecule switches. AB - New cruciform structures 1-4 were synthesized to investigate a new single molecule switching mechanism arising from the interplay between the molecule and the electrode surface. These molecular cruxes consist of two rod-type substructures, namely an oligophenylenevinylene and an oligophenyleneethynyl. While the oligophenylenevinylene rods are functionalized with acetyl protected sulfur anchor groups, the oligophenyleneethynyl rods provide terminal pyridine units. The hypothesized switching mechanism should arise from the electrochemical potential dependent coordination of the pyridine unit to the electrode surface. The assembly of the oligophenylenevinylene substructure was based on a Wittig reaction whereas its perpendicular oligophenyleneethynyl rod was assembled by Sonogashira-Hagihara coupling reactions. Preliminary transport investigations with molecular cruciforms 2 and 4 in a mechanical controllable break junction in a liquid environment displayed the trapping of single molecules between two gold electrodes via the terminally sulfur functionalized oligophenylenevinylene rod. PMID- 17915928 TI - A practical synthesis of differentially protected 2-(hydroxymethyl)piperazines. AB - An efficient and scalable synthesis of three differentially protected 2 (hydroxymethyl)piperazines is presented, starting from optically active and commercially available (2S)-piperazine-2-carboxylic acid dihydrochloride. These synthetic building blocks are useful in the preparation of biologically active compounds and as chemical scaffolds for the construction of combinatorial libraries. PMID- 17915929 TI - Chemoselective activation of nucleoside 3'-O-methylphosphonothioates with 1,3,5 triazinyl morpholinium salts. AB - Chemoselective and stereospecific O-activation of 2'-deoxynucleoside 3'-O methylphosphonothioates 5 with N-methyl-N-4,6-dimethoxy-1,3,5-triazin-2-yl morpholinium salts results in formation with retention of configuration of 5'-O DMT-2'-deoxynucleoside 3'-O-(4,6 dimethoxy-1,3,5-triazin-2-yl methylphosphonothioates (7). Active esters 7 are convenient intermediates for hydrolytic interconversion of RP-5 into SP-5 and can be used as monomers for stereoselective synthesis of dinucleoside (3',5')-methyl phosphonothioates. PMID- 17915930 TI - Vilsmeier-Haack reactions of 2-arylamino-3-acetyl-5,6-dihydro-4H-pyrans toward the synthesis of highly substituted pyridin-2(1H)-ones. AB - A facile and efficient one-pot synthesis of highly substituted pyridin-2(1H)-ones was developed via Vilsmeier-Haack reactions of readily available enaminones, 2 arylamino-3-acetyl-5,6-dihydro-4H-pyrans, and a mechanism involving sequential ring-opening, haloformylation, and intramolecular nucleophilic cyclization reactions is proposed. PMID- 17915931 TI - Scope of the Suzuki-Miyaura aminoethylation reaction using organotrifluoroborates. AB - Potassium beta-aminoethyltrifluoroborates were prepared in good yields via hydroboration of the corresponding enecarbamates using the Snieckus hydroborating reagent. A wide variety of phenethylamines containing a potentially free primary amine after appropriate deprotection have been successfully prepared in good yield using these organotrifluoroborates as partners in Suzuki-Miyaura coupling with aryl bromides, iodides, and triflates. PMID- 17915932 TI - O-attack versus N-attack: electrophilic halocyclization of unsaturated amides with vinylic halogen substitution. AB - Electrophilic iodocyclization of unsaturated amides with an internal vinylic halogen (Cl, Br, or I) substitution afforded exclusively the corresponding cyclic iminoketones via iodolactamization. On the other hand, amides having a terminal vinylic halogen substituent underwent iodolactonization only. Theoretical calculations revealed that the iodocyclization proceeds via the intramolecular iodonium ion transfer from the amide nitrogen to the C=C double bond. PMID- 17915933 TI - Catalytic asymmetric synthesis of alpha-amino acids. PMID- 17915934 TI - Spin-orbit coupling induced interference in quantum corrals. AB - Lack of inversion symmetry at a metallic surface can lead to an observable spin orbit interaction. For certain metal surfaces, such as the Au(111) surface, the experimentally observed spin-orbit coupling results in spin rotation lengths on the order of tens of nanometers, which is the typical length scale associated with quantum corral structures formed on metal surfaces. In this work, multiple scattering theory is used to calculate the local density of states (LDOS) of quantum corral structures composed of nonmagnetic adatoms in the presence of spin orbit coupling. Contrary to previous theoretical predictions, spin-orbit coupling induced modulations are observed in the theoretical LDOS, which should be observable using scanning tunneling microscopy. PMID- 17915936 TI - Copolymers including L-histidine and hydrophobic moiety for preparation of nonbiofouling surface. AB - A new type of copolymer composed of l-histidine (ampholyte) and n-butyl methacrylate (hydrophobic moiety) was developed for the preparation of nonbiofouling surfaces. The copolymer adsorbed onto resin surfaces and made the surface very hydrophilic. The hydrophilization effect was higher than that of bovine serum albumin (BSA). When polystyrene surfaces were coated with the copolymer, both the nonspecific adsorption of protein and the adhesion of cells were significantly reduced in comparison with BSA coating. The newly synthesized polymer is a new and useful candidate for the preparation of nonbiofouling surfaces. PMID- 17915937 TI - Statistics of single-molecule surface enhanced Raman scattering signals: fluctuation analysis with multiple analyte techniques. AB - The mathematical background, based on a variation of the principal component analysis (PCA) method, is developed for the understanding of fluctuating multiple analyte single-molecule (SM) surface enhanced Raman scattering (SERS) signals; with emphasis on the bianalyte SERS technique developed recently. The method and its significance are presented to provide a systematic framework with which several aspects of the statistics of SM-SERS signals can be analyzed in general. We also apply the method to a concrete example of bianalyte statistics in silver colloidal solutions and discuss related topics around experimental issues and the interpretation of single-molecule SERS data. PMID- 17915935 TI - Peptide-functionalized poly(ethylene glycol) star polymers: DNA delivery vehicles with multivalent molecular architecture. AB - Exploring the development of nonviral nucleic acid delivery vectors with progressive, specific, and novel designs in molecular architecture is a fundamental way to investigate how aspects of chemical and physical structure impact the transfection process. In this study, macromolecules comprised of a four-arm star poly(ethylene glycol) and termini modified with one of five different heparin binding peptides have been investigated for their ability to bind, compact, and deliver DNA to mammalian cells in vitro. These new delivery vectors combine a PEG-derived stabilizing moiety with peptides that exhibit unique cell-surface binding ability in a molecular architecture that permits multivalent presentation of the cationic peptides. Five peptide sequences of varying heparin binding affinity were studied; each was found to sufficiently bind heparin for biological application. Additionally, the macromolecules were able to bind and compact DNA into particles of proper size for endocytosis. In biological studies, the PEG-star peptides displayed a range of toxicity and transfection efficiency dependent on the peptide identity. The vectors equipped with peptides of highest heparin binding affinity were found to bind DNA tightly, increase levels of cellular internalization, and display the most promising transfection qualities. Our results suggest heparin binding peptides with specific sequences hold more potential than nonspecific cationic polymers to optimize transfection efficiency while maintaining cell viability. Furthermore, the built-in multivalency of these macromolecules may allow simultaneous binding of both DNA at the core of the polyplex and heparan sulfate on the surface of the cell. This scheme may facilitate a bridging transport mechanism, tethering DNA to the surface of the cell and subsequently ushering therapeutic nucleic acids into the cell. This multivalent star shape is therefore a promising architectural feature that may be exploited in the design of future polycationic gene delivery vectors. PMID- 17915938 TI - Development and application of C60-fullerene bound silica for solid-phase extraction of biomolecules. AB - Sample pretreatment is the most important procedure to remove the matrix for interfacing with mass spectrometry (MS). Additionally, for the samples with low concentration, the process of preconcentration is required before MS analysis. We have newly developed a solid-phase extraction stationary phase based on C60 fullerene covalently bound to silica for purification of biomolecules of different characteristics. Silica particles of different porosity are modified with aminopropyl linker and then covalently bound to C60-fullerenoacetic acid or C60-epoxyfullerenes. The developed materials have been successfully applied as an alternative to commercially available reversed-phase materials for solid-phase extraction. C60-fullerene silica is able to retain small and hydrophilic molecules like phosphopeptides, which can be easily lost by reversed-phase sorbents. The novel materials are applied for desalting and preconcentration of proteins and peptides, especially phosphopeptides. In addition, the C60-fullerene silica is applied for the solid-phase extraction of selected flavonoids with recoveries of approximately 99%. The recoveries are compared with the commercially available solid-phase extraction materials. PMID- 17915939 TI - Determination of surface selection rule of surface plasmon resonance near infrared spectroscopy by using a Langmuir-Blodgett film. AB - Near-infrared (NIR) absorption spectra of a cadmium arachidate Langmuir-Blodgett (LB) film were measured by surface plasmon resonance near-infrared spectroscopy (SPR-NIRS) based on the Kretschmann configuration with a 18.8-nm gold film. An NIR spectrum enhanced severalfold was obtained as a top ridge of the SPR-NIR spectra measured at different incident angles by using the principle of absorption-sensitive SPR. In order to determine the surface selection rule of SPR NIRS, the enhanced NIR absorption spectrum of the LB film was compared to an unenhanced one without the gold film and to a normal incidence transmission spectrum. Moreover, a pair of out-of-plane (OP) and in-plane (IP) spectra were obtained by multiangle infrared spectroscopy analysis from a series of oblique incidence transmission measurements in the NIR region. It became obvious that the salient feature of the enhanced NIR absorption spectrum, i.e., the top ridge of the SPR-NIR spectra is almost equivalent to that of the OP spectrum. On the other hand, the unenhanced spectrum showed IP modes. These experimental results were well explained by calculation of the mean-square electric field based on the Fresnel formula. PMID- 17915941 TI - Conformational dynamics of DNA polymerase probed with a novel fluorescent DNA base analogue. AB - DNA polymerases discriminate between correct and incorrect nucleotide substrates during a "nonchemical" step that precedes phosphodiester bond formation in the enzymatic cycle of nucleotide incorporation. Despite the importance of this process in polymerase fidelity, the precise nature of the molecular events involved remains unknown. Here we report a fluorescence resonance energy transfer (FRET) system that monitors conformational changes of a polymerase-DNA complex during selection and binding of nucleotide substrates. This system utilizes the fluorescent base analogue 1,3-diaza-2-oxophenothiazine (tC) as the FRET donor and Alexa-555 (A555) as the acceptor. The tC donor was incorporated within a model DNA primer/template in place of a normal base, adjacent to the primer 3' terminus, while the A555 acceptor was attached to an engineered cysteine residue (C751) located in the fingers subdomain of the Klenow fragment (KF) polymerase. The FRET efficiency increased significantly following binding of a correct nucleotide substrate to the KF-DNA complex, showing that the fingers had closed over the active site. Fluorescence anisotropy titrations utilizing tC as a reporter indicated that the DNA was more tightly bound by the polymerase under these conditions, consistent with the formation of a closed ternary complex. The rate of the nucleotide-induced conformational transition, measured in stopped flow FRET experiments, closely matched the rate of correct nucleotide incorporation, measured in rapid quench-flow experiments, indicating that the conformational change was the rate-limiting step in the overall cycle of nucleotide incorporation for the labeled KF-DNA system. Taken together, these results indicate that the FRET system can be used to probe enzyme conformational changes that are linked to the biochemical function of DNA polymerase. PMID- 17915943 TI - Dynamic strength of the interaction between lung surfactant protein D (SP-D) and saccharide ligands. AB - In order to investigate the dynamic strength of the interaction between lung surfactant protein D (SP-D) and different sugars, maltose, mannose, glucose, and galactose, we have used an atomic force microscope to monitor the interaction on a single molecule scale. The experiment is performed by measuring the rupture force when the SP-D-sugar bond is subjected to a continuously increasing force. Under these dynamic conditions, SP-D binds strongest to d-mannose and weakest to maltose and d-galactose. These results differ from equilibrium measurements wherein SP-D exhibits preference for maltose. On the basis of this finding, we propose that the binding of the disaccharide maltose to SP-D, which is energetically stronger than the binding of any of the monosacchrides, alters the structure of the binding site in a way that lowers the dynamic strength of the bond. We conclude that determining the strength of a protein-ligand bond under dynamic stress using an atomic force microscope is possibly more relevant for mimicking the actual nonequilibrium physiological situation in the lungs. PMID- 17915942 TI - Solution structure of polymerase mu's BRCT Domain reveals an element essential for its role in nonhomologous end joining. AB - The solution structure and dynamics of the BRCT domain from human DNA polymerase mu, implicated in repair of chromosome breaks by nonhomologous end joining (NHEJ), has been determined using NMR methods. BRCT domains are typically involved in protein-protein interactions between factors required for the cellular response to DNA damage. The pol mu BRCT domain is atypical in that, unlike other reported BRCT structures, the pol mu BRCT is neither part of a tandem grouping, nor does it appear to form stable homodimers. Although the sequence of the pol mu BRCT domain has some unique characteristics, particularly the presence of >10% proline residues, it forms the characteristic alphabetaalpha sandwich, in which three alpha helices are arrayed around a central four-stranded beta-sheet. The structure of helix alpha1 is characterized by two solvent-exposed hydrophobic residues, F46 and L50, suggesting that this element may play a role in mediating interactions of pol mu with other proteins. Consistent with this argument, mutation of these residues, as well as the proximal, conserved residue R43, specifically blocked the ability of pol mu to efficiently work together with NHEJ factors Ku and XRCC4-ligase IV to join noncomplementary ends together in vitro. The structural, dynamic, and biochemical evidence reported here identifies a functional surface in the pol mu BRCT domain critical for promoting assembly and activity of the NHEJ machinery. Further, the similarity between the interaction regions of the BRCT domains of pol mu and TdT support the conclusion that they participate in NHEJ as alternate polymerases. PMID- 17915944 TI - Caulollins from Caulobacter crescentus, a pair of partially unstructured proteins of betagamma-crystallin superfamily, gain structure upon binding calcium. AB - The betagamma-crystallin superfamily comprises members from various taxa and species, which have similar domain topologies as that of lens beta- and gamma crystallins. We have studied new microbial members of this understudied betagamma crystallin superfamily from the bacterium Caulobacter crescentus. These proteins, which we named "caulollins", are paralogues with a single betagamma-crystallin domain, made up of two Greek key motifs with AB-type arrangement seen in gamma crystallin. The second Greek key motif has Cys in place of a generally conserved Phe/Tyr residue, and the Tyr corner, considered important for the proper betagamma-crystallin fold, is missing, making this a sequentially diverse atypical betagamma-crystallin domain. This atypical domain binds two Ca2+ with moderate affinity (0.8-20 microM). In apo form, caulollins are partially unstructured proteins and gain structure upon binding Ca2+. Unlike many other microbial betagamma-crystallin domains, this domain is monomeric, though in the presence of Ca2+ it becomes more compact. Ca2+ binding increases the intrinsic stability of proteins, suggesting the role of Ca2+ as an extrinsic stabilizer. N Terminal extension does not play any role in modulating Ca2+ binding, intrinsic stability, or oligomerization. We noted that there are several such variant domains in the genomes of unrelated species. It appears that caulollins along with these members form a subfamily in the betagamma-crystallin superfamily that would be partially unstructured in apo form, unlike many other domains from lens or microbial crystallins. This work further suggests that Ca2+ binding is a widespread feature of the betagamma-crystallin superfamily. PMID- 17915945 TI - The N-terminal (1-44) and C-terminal (198-243) peptides of apolipoprotein A-I behave differently at the triolein/water interface. AB - Apolipoprotein A-I (apoA-I), the major protein of high-density lipoprotein (HDL), moves between HDL and triacylglycerol-rich lipoproteins during metabolism. We reported that apoA-I is conformationally flexible at the triolein/water (TO/W) interface, partially desorbing at low surface pressure (Pi) but totally desorbing at Pi > 19 mN/m. We now report the different behavior of the N- and C-terminal peptides of apoA-I ([1-44]apoA-I and [198-243]apoA-I) at the TO/W interface. While both peptides are surface active, [198-243]apoA-I is more stable at the TO/W interface. At equilibrium interfacial tension both peptides desorb from the interface when compressed, but [1-44]apoA-I is pushed off at 13 mN/m while [198 243]apoA-I can withstand Pi = 16 mN/m. Neither peptide is very elastic or flexible at the interface. Only at small changes of area (<8%), fast oscillations (4 and 8 s periods), and relatively low concentrations (2 x 10(-7) M) do these peptides show elastic behavior but with a relatively small modulus compared to that of apoA-I. When mixed together, they appear not to interact on the surface. [1-44]ApoA-I binds more rapidly but is replaced by [198-243]apoA-I within minutes. We suggest that when apoA-I partially desorbs from lipoprotein surfaces during lipid metabolism, the N-terminal is the first to detach while the C terminal remains on the interface and only desorbs at higher pressures. Thus, the observations that different domains of apoA-I adsorb or desorb with small variations in surface pressure make apoA-I a very flexible protein with multiple functions, one of which is to stabilize surface pressure during lipoprotein metabolism as lipids move in and out of the lipoprotein surface. PMID- 17915946 TI - Thermodynamic analysis of allosamidin binding to a family 18 chitinase. AB - Inhibition of family 18 chitinases is emerging as a target for pest and fungal control as well as asthma and inflammatory therapy. One of the best known inhibitors for these enzymes is allosamidin, a natural product. While interactions of this compound with family 18 chitinases have been studied in much detail by X-ray crystallography and standard enzymology, details of the driving forces behind its tight binding remain unknown. We have studied the thermodynamics of allosamidin binding to chitinase B (ChiB), a family 18 chitinase from Serratia marcescens, using isothermal titration calorimetry. At pH 6.0, Kd is 0.16 +/- 0.04 microM, and the binding reaction is entropically driven (DeltaSr = 44 cal/K mol) with an enthalpic penalty (DeltaHr = 3.8 +/- 0.2 kcal/mol). Dissection of the entropic term shows that a favorable conformational change in the allosamidin-ChiB complex (DeltaSconf = 37 cal/K mol) is the main contributor to the reaction. At pH 8.5, Kd decreases to 0.03 muM and the binding reaction is less entropically favorable (DeltaSr = 30 cal/K mol). While the solvation entropy change (DeltaSsolv) increases from 15 cal/K mol at pH 6.0 to 46 cal/K mol at pH 8.5, DeltaSconf becomes small and negative (-8 cal/K mol) because of an enthalpy-entropy compensation. Analyses of proton transfer showed that at pH 6.0 binding of allosamidin requires deprotonation of the Asp142-Glu144 catalytic diad. At pH 8.5, the 142-144 diad is ionized in the native enzyme, relieving the deprotonation penalty of binding and explaining why binding becomes enthalpically favorable (DeltaHr = -1.2 +/- 0.2 kcal/mol). PMID- 17915947 TI - Transmembrane domain of myelin protein zero can form dimers: possible implications for myelin construction. AB - Myelin protein zero (MPZ) is the major integral membrane protein of peripheral nerve myelin in higher vertebrates, mediating homoadhesion of the multiple, spiraling wraps of the myelin sheath. Previous studies have shown that full length MPZ can form dimers and tetramers, and biochemical studies on the extracellular domain (ECD) indicate that it can form a tetramer, albeit very weakly. On the basis of cross-linking studies and equilibrium sedimentation of a transmembrane (TM) domain peptide (MPZ-TM), we find that the MPZ-TM can form homodimers. We further characterized the dimer by measuring the effects of alanine and leucine substitutions on the ability of the TM to dimerize in Escherichia coli membranes. Our results indicate that the primary packing interface for the MPZ TM homodimer is a glycine zipper (GxxxGxxxG) motif. We also find that the G134R mutation, which lies within the glycine zipper packing interface and causes Charcot-Marie-Tooth disease type 1B, severely inhibits dimerization, suggesting that dimerization of the TM domain may be important for the normal functioning of MPZ. By combining our new results with prior work, we suggest a new model for an MPZ lattice that may form during the construction of myelin. PMID- 17915948 TI - Thermodynamics of b-HLH-LZ protein binding to DNA: the energetic importance of protein-DNA contacts in site-specific E-box recognition by the complete gene product of the Max p21 transcription factor. AB - The Myc/Mad/Max network of dimeric basic region-helix-loop-helix-leucine zipper (b-HLH-LZ) transcription factors bind to enhancer box sequences (E-box) in the promotors of a large set of genes that control cell metabolism, proliferation, and differentiation. Max (Myc-associated factor X) is the obligate heterodimerization partner of Myc and Mad proteins. On the other hand, Max is the only member of the family capable of forming a stable homodimer. As part of the transcriptional regulation mechanism, Myc/Max and Mad/Max heterodimers and Max homodimers are thought to compete for binding to the E-box target sequences. E box recognition is structurally supported by the b-HLH-LZ structural motif, which also promotes dimerization. However, the actual dimerization and heterodimerization constants of the complete gene products and their affinities for E-box sequences are not known. Also, the detailed thermodynamic characterization of DNA binding by these transcription factors has not been done yet. Such knowledge is necessary for complete understanding of the transcriptional regulation carried out by the Myc/Mad/Max network. Here, we report the first in-depth thermodynamic characterization of the stability and specific DNA binding of a full length gene product of the Myc/Mad/Max family, namely, Max protein isoform p21 (Max p21). Using calorimetric methods (DSC and ITC) we have determined the dimerization constant of Max p21 in the low micromolar range, and the Max p21/E-box complex dissociation constant in the low nanomolar range at 37 degrees C. The association is driven by a large exothermic effect, which is partly compensated by entropic factors. The energetic contribution to binding affinity of seven highly conserved residues that contact the DNA was probed by X-to-Ala mutagenesis. The results demonstrate that high binding affinity critically relies on the side chain of Arg 26. Furthermore, the mutational analysis points to the important role of the persistent helical turn that comprises this residue at the junction of the basic region and helix H1. Altogether, the study supports the idea that Max p21 can bind E-box sequences in vivo and likely participates directly in the regulation of transcription as homodimer. PMID- 17915949 TI - Increased stability and DNA site discrimination of "single chain" variants of the dimeric beta-barrel DNA binding domain of the human papillomavirus E2 transcriptional regulator. AB - Human papillomavirus infects millions of people worldwide and is a causal agent of cervical cancer in women. The HPV E2 protein controls the expression of all viral genes through binding of its dimeric C-terminal domain (E2C) to its target DNA site. We engineered monomeric versions of the HPV16 E2C, in order to probe the link of the dimeric beta-barrel fold to stability, dimerization, and DNA binding. Two single-chain variants, with 6 and 12 residue linkers (scE2C-6 and scE2C-12), were purified and characterized. Spectroscopy and crystallography show that the native structure is unperturbed in scE2C-12. The single chain variants are stabilized with respect to E2C, with effective concentrations of 0.6 to 6 mM. The early folding events of the E2C dimer and scE2C-12 are very similar and include formation of a compact species in the submillisecond time scale and a non native monomeric intermediate with a half-life of 25 ms. However, monomerization changes the unfolding mechanism of the linked species from two-state to three state, with a high-energy intermediate. Binding to the specific target site is up to 5-fold tighter in the single chain variants. Nonspecific DNA binding is up to 7-fold weaker in the single chain variants, leading to an overall 10-fold increased site discrimination capacity, the largest described so far for linked DNA binding domains. Titration calorimetric binding analysis, however, shows almost identical behavior for dimer and single-chain species, suggesting very subtle changes behind the increased specificity. Global analysis of the mechanisms probed suggests that the dynamics of the E2C domain, rather than the structure, are responsible for the differential properties. Thus, the plastic and dimeric nature of the domain did not evolve for a maximum affinity, specificity, and stability of the quaternary structure, likely because of regulatory reasons and for roles other than DNA binding played by partly folded dimeric or monomeric conformers. PMID- 17915950 TI - Biochemical and structural characterization of Pseudomonas aeruginosa Bfd and FPR: ferredoxin NADP+ reductase and not ferredoxin is the redox partner of heme oxygenase under iron-starvation conditions. AB - Among the 118 genes upregulated by Pseudomonas aeruginosa in response to iron starvation [Ochsner, U. A., Wilderman, P. J., Vasil, A. I., and Vasil, M. L. (2002) Mol. Microbiol. 45, 1277-1287], we focused on the products of the two genes encoding electron transfer proteins, as a means of identifying the redox partners of the heme oxygenase (pa-HO) expressed under low-iron stress conditions. Biochemical and spectroscopic investigations demonstrated that the bfd gene encodes a 73-amino acid protein (pa-Bfd) that incorporates a [2Fe 2S]2+/+ center, whereas the fpr gene encodes a 258-residue NADPH-dependent ferredoxin reductase (pa-FPR) that utilizes FAD as a cofactor. In vitro reconstitution of pa-HO catalytic activity with the newly characterized proteins led to the surprising observation that pa-FPR efficiently supports the catalytic cycle of pa-HO, without the need of a ferredoxin. In comparison, electron transfer from pa-Bfd to pa-HO is sluggish, which strongly argues against the possibility that the seven electrons needed by pa-HO to degrade biliverdin are transferred from NADPH to pa-HO in a ferredoxin (Bfd)-dependent manner. Given that pa-HO functions to release iron from exogenous heme acquired under iron starvation conditions, the use of a flavoenzyme rather than an iron-sulfur center containing protein to support heme degradation is an efficient use of resources in the cell. The crystal structure of pa-FPR (1.6 A resolution) showed that its fold is comparable that of the superfamily of ferredoxin reductases and most similar to the structure of Azotobacter vinelandii FPR and Escherichia coli flavodoxin reductase. The latter two enzymes interact with distinct redox partners, a ferredoxin and a flavodoxin, respectively. Hence, findings reported herein extend the range of redox partners recognized by the fold of pa-FPR to include a heme oxygenase (pa-HO). PMID- 17915951 TI - Kinetic evidence is consistent with the rocker-switch mechanism of membrane transport by GlpT. AB - Secondary active transport of substrate across the cell membrane is crucial to many cellular and physiological processes. The crystal structure of one member of the secondary active transporter family, the sn-glycerol-3-phosphate (G3P) transporter (GlpT) of the inner membrane of Escherichia coli, suggests a mechanism for substrate translocation across the membrane that involves a rocker switch-type movement of the protein. This rocker-switch mechanism makes two specific predictions with respect to kinetic behavior: the transport rate increases with the temperature, whereas the binding affinity of the transporter to a substrate is temperature-independent. In this work, we directly tested these two predictions by transport kinetics and substrate-binding experiments, integrating the data on this single system into a coherent set of observations. The transport kinetics of the physiologically relevant G3P-phosphate antiport reaction were characterized at different temperatures using both E. coli whole cells and GlpT reconstituted into proteoliposomes. Substrate-binding affinity of the transporter was measured using tryptophan fluorescence quenching in detergent solution. Indeed, the substrate transport velocity of GlpT increased dramatically with temperature. In contrast, neither the apparent Michaelis constant (Km) nor the apparent substrate-binding dissociation constant (Kd) showed temperature dependence. Moreover, GlpT-catalyzed G3P translocation exhibited a completely linear Arrhenius function with an activation energy of 35.2 kJ mol-1 for the transporter reconstituted into proteoliposomes, suggesting that the substrate loaded transporter is delicately poised between the inward- and outward-facing conformations. When these results are taken together, they are in agreement with a rocker-switch mechanism for GlpT. PMID- 17915952 TI - Mutation of arginine 357 of the CP43 protein of photosystem II severely impairs the catalytic S-state cycle of the H2O oxidation complex. AB - Basic amino acid side chains situated in active sites may mediate critical proton transfers during an enzymatic catalytic cycle. In the case of photosynthetic water oxidation, a strong base is postulated to facilitate the deprotonation of the active site Mn4-Ca cluster, thereby allowing the otherwise thermodynamically constrained transfer of an electron away from the Mn4-Ca cluster to the oxidized redox active tyrosine radical, YZ*, generated by photosynthetic charge separation. Arginine 357 of the CP43 polypeptide may be located in the second coordination shell of the O2-evolving Mn4-Ca cluster of photosystem II (PSII) according to current structural models. An ostensibly conservative substitution mutation, CP43-357K, was investigated using polarographic and fluorescence techniques in evaluating its potential impact on S-state cycling. Cells containing the CP43-357K mutation lost their capacity for autotrophic growth and exhibited a drastic reduction in O2 evolving activity ( approximately 15% of that of the wild type) despite the fact that mutant cells contained more than 80% of the concentration of charge-separating PSII reaction centers and more than half of these contained photooxidizable Mn. Fluorescence kinetics indicated that acceptor side electron transfer, dominated by the transfer of electrons from QA- to QB, was unaffected, but the fraction of centers containing Mn clusters capable of forming the S2 state was reduced to approximately 40% of that of the wild type. Analysis of O2 yields using a bare platinum electrode indicated a severe defect in the S-state cycling properties of the mutant H2O oxidation complexes. Although O2 evolution was delayed to the third flash during a train of single turnover saturating flashes, the pattern of O2 emission did not exhibit a discernible periodicity indicating a very high miss factor, which was estimated to be approximately 45% compared to the wild-type value of approximately 10%. On the other hand, the multiflash fluorescence measurements indicate that the yield of formation of the S2 state from S1 is diminished by approximately 20%, although this latter estimate is complicated by the presence of damaged PSII centers. Taken together, the experiments indicate that the high miss factor observed during S-state cycling is likely due to a defect in the higher S-state transitions. These results are discussed in relation to the idea that CP43-R357 may serve as a ligand to bicarbonate or as the catalytic base proposed to mediate proton-coupled electron transfer (PCET) in the higher S states of the catalytic cycle of H2O oxidation. PMID- 17915954 TI - Irreversible inhibition of the Mycobacterium tuberculosis beta-lactamase by clavulanate. AB - Members of the beta-lactam class of antibiotics, which inhibit the bacterial d,d transpeptidases involved in cell wall biosynthesis, have never been used systematically in the treatment of Mycobacterium tuberculosis infections because of this organism's resistance to beta-lactams. The critical resistance factor is the constitutive production of a chromosomally encoded, Ambler class A beta lactamase, BlaC in M. tuberculosis. We show that BlaC is an extended spectrum beta-lactamase (ESBL) with high levels of penicillinase and cephalosporinase activity as well as measurable activity with carbapenems, including imipenem and meropenem. We have characterized the enzyme's inhibition by three FDA-approved beta-lactamase inhibitors: sulbactam, tazobactam, and clavulanate. Sulbactam inhibits the enzyme competitively and reversibly with respect to nitrocefin. Tazobactam inhibits the enzyme in a time-dependent manner, but the activity of the enzyme reappears due to the slow hydrolysis of the covalently acylated enzyme. In contrast, clavulanate reacts with the enzyme quickly to form hydrolytically stable, inactive forms of the enzyme that have been characterized by mass spectrometry. Clavulanate has potential to be used in combination with approved beta-lactam antibiotics to treat multi-drug resistant (MDR) and extremely drug resistant (XDR) strains of M. tuberculosis. PMID- 17915953 TI - Expression and characterization of full-length human heme oxygenase-1: the presence of intact membrane-binding region leads to increased binding affinity for NADPH cytochrome P450 reductase. AB - Heme oxygenase-1 (HO-1) is the chief regulatory enzyme in the oxidative degradation of heme to biliverdin. In the process of heme degradation, HO-1 receives the electrons necessary for catalysis from the flavoprotein NADPH cytochrome P450 reductase (CPR), releasing free iron and carbon monoxide. Much of the recent research involving heme oxygenase has been done using a 30 kDa soluble form of the enzyme, which lacks the membrane binding region (C-terminal 23 amino acids). The goal of this study was to express and purify a full-length human HO-1 (hHO-1) protein; however, due to the lability of the full-length form, a rapid purification procedure was required. This was accomplished by use of a glutathione-s-transferase (GST)-tagged hHO-1 construct. Although the procedure permitted the generation of a full-length HO-1, this form was contaminated with a 30 kDa degradation product that could not be eliminated. Therefore, attempts were made to remove a putative secondary thrombin cleavage site by a conservative mutation of amino acid 254, which replaces arginine with lysine. This mutation allowed the expression and purification of a full-length hHO-1 protein. Unlike wild type (WT) HO-1, the R254K mutant could be purified to a single 32 kDa protein capable of degrading heme at the same rate as the WT enzyme. The R254K full-length form had a specific activity of approximately 200-225 nmol of bilirubin h-1 nmol-1 HO-1 as compared to approximately 140-150 nmol of bilirubin h-1 nmol-1 for the WT form, which contains the 30 kDa contaminant. This is a 2-3 fold increase from the previously reported soluble 30 kDa HO-1, suggesting that the C-terminal 23 amino acids are essential for maximal catalytic activity. Because the membrane-spanning domain is present, the full-length hHO-1 has the potential to incorporate into phospholipid membranes, which can be reconstituted at known concentrations, in combination with other endoplasmic reticulum resident enzymes. PMID- 17915955 TI - Development of albumin-binding camptothecin prodrugs using a Peptide positional scanning library. AB - Designing truly tumor-specific prodrugs remains a challenge in the field of cancer chemotherapy. As a new strategy, we incubated homogenates of a spectrum of human colon tumor xenografts with a fluorogenic positional scanning tetrapeptide library in order to identify peptide sequences that are preferentially cleaved by colon tumors. Our screening experiments at pH 7.4 revealed that Met, Leu, and Lys were preferred amino acids in the position P(1) and Tyr, Phe, and Met in P(2), whereas in P(3) and P(4), the cleavage profiles were less characteristic. However, similar results were obtained when testing breast tumor material and homogenates from healthy murine organs. On the basis of these results, we developed albumin-binding camptothecin (CPT) prodrugs of the general formula EMC Arg-P(4)-P(3)-P(2)-P(1)-Ala-CPT (EMC = 6-maleimidocaproic acid) that incorporated two peptide linkers: H-Arg-Ala-Phe-Met-OH and H-Arg-Phe-Tyr-Met-OH (P(4)-P(3) P(2)-P(1)). The incorporation of two arginine residues rendered the prodrugs water-soluble (>7 mg/mL), while the use of alanine as an amino acid spacer proved to be beneficial for the release of the active agent. Incubation studies with homogenates of HT-29 colon tumor tissue and murine spleen, liver, and kidneys demonstrated cleavage of the peptide linker with CPT-peptide derivatives and CPT being the major cleavage products. Although the peptide sequence is not selectively cleaved in colon tumors, an in vivo study in a HT-29 xenograft model showed that the prodrug EMC-Arg-Arg-Ala-Phe-Met-Ala-CPT demonstrated enhanced antitumor efficacy when compared to CPT [( T/ C max: 17% for the prodrug (2 x 12.5 mg/kg CPT equivalents) and 40% for CPT (3 x 12.5 mg/kg)]. PMID- 17915956 TI - Targeting lipopolyplexes using bifunctional peptides incorporating hydrophobic spacer amino acids: synthesis, transfection, and biophysical studies. AB - We have developed efficient synthetic routes to two hydrophobic amino acids, suitably protected for solid-phase peptide synthesis, and have successfully synthesized peptides containing these or other hydrophobic amino acids as spacers between a Lys16 moiety and an integrin-targeting motif. These peptides have in turn been used to formulate a range of lipopolyplex vectors with Lipofectin and plasmid DNA. The transfection efficiencies of these vectors and their aggregation behavior in buffers and in serum have been studied. We have shown that vectors containing peptides incorporating long linkers that are entirely hydrophobic are less efficient transfection agents. However, linkers of equivalent length that are in part hydrophobic show improved transfection properties, which is probably due to the improved accessibility of the integrin-binding motif. PMID- 17915957 TI - Facile and rapid direct gold surface immobilization with controlled orientation for carbohydrates. AB - Effective surface immobilization is a prerequisite for numerous carbohydrate related studies including carbohydrate-biomolecule interactions. In the present work, we report a simple and rapid modification technique for diverse carbohydrate types in which direct oriented immobilization onto a gold surface is accomplished by coupling the amine group of a thiol group-bearing aminophenyl disulfide as a new coupling reagent with an aldehyde group of the terminal reducing sugar in the carbohydrate. To demonstrate the generality of this proposed reductive amination method, we examined its use for three types of carbohydrates: glucose (monosaccharide), lactose (disaccharide), and GM1 pentasaccharide. Through successful mass identifications of the modified carbohydrates, direct binding assays on gold surface using surface plasmon resonance and electrochemical methods, and a terminal galactose-binding lectin assay using atomic force microscopy, we confirmed several advantages including direct and rapid one-step immobilization onto a gold surface and exposure of functional carbohydrate moieties through oriented modification of the terminal reducing sugar. Therefore, this facile modification and immobilization method can be successfully used for diverse biomimetic studies of carbohydrates, including carbohydrate-biomolecule interactions and carbohydrate sensor or array development for diagnosis and screening. PMID- 17915958 TI - Protease-sensitive fluorescent nanofibers. AB - We report the design and synthesis of enzyme-responsive nanofibers. The fibers are composed of self-assembled hydrophobic beta-sheet peptides incorporating protease-sensitive domains, fluorescent reporters, and hydrophilic poly(ethylene glycol) (PEG) units. Using urokinase plasminogen activator (uPA) as a model system, nanofibers were developed to release fluorescent fragments upon uPA incubation. These protease-sensitive nanofibers may have considerable biomedical applications as diagnostic sensors or for protease-assisted drug deliveries. PMID- 17915959 TI - Kinetic behavior of Fe(o,o-EDDHA)-humic substance mixtures in several soil components and in calcareous soils. AB - Ferric ethylenediamine- N, N'-bis-(o-hydroxyphenylacetic)acid chelate (Fe(o, o EDDHA)) is one of the most effective Fe fertilizers in calcareous soils. However, humic substances are occasionally combined with iron chelates in drip irrigation systems in order to lower costs. The reactivity of iron chelate-humic substance mixtures in several soil components and in calcareous soils was investigated through interaction tests, and their behavior was compared to the application of iron chelates and humic substances separately. Two commercial humic substances and two Fe(o, o-EDDHA) chelates (one synthesized in the laboratory and one commercial) were used to prepare iron chelate-humic substance mixtures at 50% (w/w). Various soil components (calcium carbonate, gibbsite, amorphous iron oxide, hematite, tenorite, zincite, amorphous Mn oxide, and peat) and three calcareous soils were shaken for 15 days with the mixtures and with iron chelate and humic substance solutions. The kinetic behavior of Fe(o, o-EDDHA) and Fe non (o,o-EDDHA) (Fe bonded to (o,p-EDDHA) and other polycondensated ligands) and of the different nutrients solubilized after the interaction assay was determined. The results showed that the mixtures did not significantly reduce the retention of Fe(o, o-EDDHA) and Fe non-(o,o-EDDHA) in the soil components and the calcareous soils compared to the iron chelate solutions, but they did produce changes in the retention rate. Moreover, the competition between humic substances and synthetic chelating agents for complexing metal cations limited the effectiveness of the mixtures to mobilize nutrients from the substrates. The presence of Fe(o, p-EDDHA) and other byproducts in the commercial iron chelate had an important effect on the evolution of Fe(o, o-EDDHA) and the nutrient solubilization process. PMID- 17915960 TI - Glycoprofiling of bifidobacterial consumption of human milk oligosaccharides demonstrates strain specific, preferential consumption of small chain glycans secreted in early human lactation. AB - The molecular basis by which human breast milk supports the development of a protective intestinal microbiome in infants is unknown. After lactose and lipids, human milk oligosaccharides (HMOs) are quantitatively the third largest and most diverse component of breast milk. In this work, glycomic profiling of HMO consumption by bifidobacteria using Fourier transform ion cyclotron resonance mass spectrometry reveals that one species, Bifidobacterium longum biovar infantis ATCC 15697, an isolate from the infant gut, preferentially consumes small mass oligosaccharides, representing 63.9% of the total HMOs available. These HMOs were detected in human breast milk at the onset and constantly through the first month of lactation by use of high performance liquid chromatography chip time-of-flight mass spectrometry. Further characterization revealed that strain ATCC 15697 possesses both fucosidase and sialidase activities not present in the other tested strains. This work provides evidence that these small mass HMOs are selectively metabolized by select bifidobacterial strains and represent a potential new class of bioactive molecules functioning as prebiotics to facilitate a protective gut colonization in breast-fed newborns. PMID- 17915961 TI - Arsenite-enhanced procoagulant activity through phosphatidylserine exposure in platelets. AB - Numerous epidemiological studies have reported the close relationship between arsenic in drinking water and cardiovascular disease (CVD), yet the exact mechanism underlying this relationship remains unclear. We investigated whether arsenic can affect the procoagulant activity of platelets, which are essential in blood clotting, thrombus formation, and progression of CVD. While arsenite alone did not induce procoagulant activity, it significantly enhanced thrombin-induced procoagulant activity of human platelets in a concentration- and time-dependent manner. In flow cytometric analysis, arsenite potentiated phosphatidylserine (PS) exposure and microparticle (MP) formation, the major mediators of procoagulant activity. Arsenite-enhanced calcium increase and subsequent calpain activation were found to be involved in these effects, as determined by confocal microscopy and gel electrophoresis. Arsenite also inhibited flippase, an enzyme that restores PS to the inner leaflet, suggesting that PS could be retained in outer membranes after exposure. Consistent with these in vitro results, ex vivo studies revealed that PS exposure in platelets was significantly increased after acute or chronic arsenic exposure in rats. Most notably, in a rodent in vivo venous thrombosis model, arsenite indeed led to increased thrombus formation. In conclusion, arsenite-enhanced procoagulant activity in platelets by PS exposure and MP generation ultimately results in accelerated thrombus formation in vivo, suggesting that this enhanced activity is a possible contributing factor in CVD associated with chronic exposure to arsenic through drinking water. PMID- 17915962 TI - Parallel synthesis of 1,3-dihydro-1,4-benzodiazepine-2-ones employing catch and release. AB - An efficient solid-phase method has been developed for the parallel synthesis of 1,3-dihydro-1,4-benzodiazepine-2-one derivatives. A key step in this procedure involves catching crude 2-aminobenzoimine products 4 on an amino acid Wang resin 10. Mild acidic conditions then promote a ring closure and in the same step cleavage from the resin to give pure benzodiazepine products 12. The 2 aminobenzoimines 4 can be synthesized from either 2-aminobenzonitriles 1 and Grignard reagents 2 or from iodoanilines 5 and nitriles 7 allowing a range of diversification. Further diversification can be introduced to the benzodiazepine products by N-alkylation promoted by a resin bound base and alkylating agents 13. PMID- 17915963 TI - Top-down proteomics on a chromatographic time scale using linear ion trap fourier transform hybrid mass spectrometers. AB - Proteomics has grown significantly with the aid of new technologies that consistently are becoming more streamlined. While processing of proteins from a whole cell lysate is typically done in a bottom-up fashion utilizing MS/MS of peptides from enzymatically digested proteins, top-down proteomics is becoming a viable alternative that until recently has been limited largely to offline analysis by tandem mass spectrometry. Here we describe a method for high resolution tandem mass spectrometery of intact proteins on a chromatographic time scale. In a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) run, we have identified 22 yeast proteins with molecular weights from 14 to 35 kDa. Using anion exchange chromatography to fractionate a whole cell lysate before online LC-MS/MS, we have detected 231 metabolically labeled (14N/15N) protein pairs from Saccharomyces cerevisiae. Thirty-nine additional proteins were identified and characterized from LC-MS/MS of selected anion exchange fractions. Automated localization of multiple acetylations on Histone H4 was also accomplished on an LC time scale from a complex protein mixture. To our knowledge, this is the first demonstration of top-down proteomics (i.e., many identifications) on linear ion trap Fourier transform (LTQ FT) systems using high resolution MS/MS data obtained on a chromatographic time scale. PMID- 17915964 TI - NMR metabolomics of planktonic and biofilm modes of growth in Pseudomonas aeruginosa. AB - Bacteria often reside in communities where the cells have secreted sticky, polymeric compounds that allow them to attach to surfaces. This sessile lifestyle, referred to as a biofilm, affords the cells within these communities a tolerance of antibiotics and antimicrobial treatments. Biofilms of the bacterium Pseudomonas aeruginosa have been implicated in cystic fibrosis and are capable of colonizing medical implant devices, such as heart valves and catheters, where treatment of the infection often requires the removal of the infected device. This mode of growth is in stark contrast to planktonic, free floating cells, which are more easily eradicated with antibiotics. The mechanisms contributing to a biofilm's tenacity and a planktonic cell's susceptibility are just beginning to be explored. In this study, we have used a metabolomic approach employing nuclear magnetic resonance (NMR) techniques to study the metabolic distinctions between these two modes of growth in P. aeruginosa. One-dimensional 1H NMR spectra of fresh growth medium were compared with spent medium supernatants from batch and chemostat planktonic and biofilms generated in continual flow system culture. In addition, 1H high-resolution magic angle spinning NMR techniques were employed to collect 1H NMR spectra of the corresponding cells. Principal component analysis and spectral comparisons revealed that the overall metabolism of planktonic and biofilm modes of growth appeared similar for the spent media, while the planktonic and biofilm cells displayed marked differences. To determine the robustness of this technique, we prepared cell samples under slightly different preparation methods. Both techniques showed similar results. These feasibility studies show that there exist chemical differences between planktonic and biofilm cells; however, in order to identify these metabolomic differences, more extensive studies would have to be performed, including 1H-1H total correlated spectroscopy. PMID- 17915965 TI - Method for identifying nonspecific protein-protein interactions in nanoelectrospray ionization mass spectrometry. AB - The nonspecific self-association of proteins in nanoflow electrospray ionization mass spectrometry (nanoES-MS), and the influence of experimental conditions thereon, are investigated using the protein ubiquitin (Ubq) as a model system. Extents of nonspecific protein association generally increase with protein concentration and, interestingly, with decreasing ES spray potential. The extent of self-association is also sensitive to the duration of the accumulation event in an external rf hexapole. Notably, the relative abundance of metal (Na+ and K+) adducts generally increases with the size of nonspecific Ubq multimer. This result suggests that the gaseous ions of monomeric and nonspecific multimeric Ubq have, on average, different ES droplet histories, with monomer ions originating earlier in the ES process than the nonspecific multimeric complexes. This finding forms the basis for a new method for distinguishing between specific and nonspecific protein complexes in ES-MS. A reporter molecule (Mrep), which does not bind specifically to the proteins and protein complexes of interest, is added to the ES solution at high concentration. The distribution of Mrep bound nonspecifically to gaseous ions of the proteins and protein complexes, as determined from the ES mass spectrum, is used to determine whether a given protein complex originates in solution or whether it forms from nonspecific binding during the ES process. The method is demonstrated in cases where the ions of protein complexes detected by nanoES-MS originate exclusively from nonspecific association, exclusively from specific interactions in solution, or from both specific and nonspecific interactions. PMID- 17915967 TI - Managing cognitive deficits and functioning in patients with schizophrenia. PMID- 17915968 TI - Importance of a correct initial diagnosis and stabilization to avoid social and economic consequences. PMID- 17915969 TI - Double-blind, randomized, placebo-controlled trial of topiramate plus cognitive behavior therapy in binge-eating disorder. AB - OBJECTIVE: To evaluate the efficacy and tolerability of adjunctive topiramate compared to placebo in reducing weight and binge eating in obese patients with binge-eating disorder (BED) receiving cognitive-behavior therapy (CBT). METHOD: A double-blind, randomized, placebo-controlled trial of 21 weeks' duration was conducted at 4 university centers. Participants were 73 obese (body mass index >or= 30 kg/m(2)) outpatients with BED (DSM-IV criteria), both genders, and aged from 18 to 60 years. After a 2- to 5-week run-in period, selected participants were treated with group CBT (19 sessions) and topiramate (target daily dose, 200 mg) or placebo (September 2003-April 2005). The main outcome measure was weight change, and secondary outcome measures were binge frequencies, binge remission, Binge Eating Scale (BES) scores, and Beck Depression Inventory (BDI) scores. RESULTS: Repeated-measures random regression analysis revealed a greater rate of weight reduction associated with topiramate over the course of treatment (p < .001), with patients taking topiramate attaining a clinically significant weight loss (-6.8 kg) compared to patients taking placebo (-0.9 kg). Although rates of reduction of binge frequencies, BES scores, and BDI scores did not differ between groups during treatment, a greater number of patients of the topiramate plus CBT group (31/37) attained binge remission compared to patients taking placebo (22/36) during the trial (p = .03). No difference between groups was found in completion rates; 1 patient (topiramate group) withdrew for adverse effect. Paresthesia and taste perversion were more frequent with topiramate, and insomnia was more frequent with placebo (p < .05). CONCLUSIONS: Topiramate added to CBT improved the efficacy of the later, increasing binge remission and weight loss in the short run. Topiramate was well tolerated, as shown by few adverse events during treatment. CLINICAL TRIALS REGISTRATION: ClinicalTrials.gov identifier NCT00307619. PMID- 17915970 TI - Ziprasidone in treatment-resistant schizophrenia: a 52-week, open-label continuation study. AB - OBJECTIVE: To evaluate the efficacy, safety, and tolerability of long-term ziprasidone therapy in treatment-resistant schizophrenia. METHOD: This prospective, 1-year, open-label study of ziprasidone (40-160 mg/day) was conducted in subjects who had participated in a previous randomized 12-week comparison of ziprasi-done and chlorpromazine in treatment-resistant schizophrenia (DSM-III-R criteria). The clinical response of 62 subjects was evaluated (32 subjects had been on ziprasidone treatment and 30 had been on chlorpromazine treatment prior to enrollment in the continuation study). Assessments included the Positive and Negative Syndrome Scale total and subscale scores, movement disorder scales, body weight, and laboratory measures. This study was conducted from May 2000 to April 2002. RESULTS: Thirty-three subjects (53%) completed 1 year of open-label ziprasidone therapy. Ziprasidone maintained clinical improvement (no significant symptom exacerbation) in 30 of 41 subjects (73%) who responded to the initial 12-week double-blind treatment with either ziprasidone or chlorpromazine. Ziprasidone did not increase body weight and was associated with a favorable metabolic profile during the continuation study period. There were no significant changes in standard movement disorder measures from the core study baseline during long-term ziprasidone treatment. CONCLUSION: Ziprasidone was effective and well tolerated in the long-term therapy of patients with treatment-resistant schizophrenia. PMID- 17915971 TI - Diurnal mood variation in outpatients with major depressive disorder: implications for DSM-V from an analysis of the Sequenced Treatment Alternatives to Relieve Depression Study data. AB - OBJECTIVE: Diurnal mood variation (DMV) with early morning worsening is considered a classic symptom of melancholic features in The Diagnostic and Statistical Manual of Mental Disorders (DSM) as well as The International Classification of Diseases (ICD) criteria for somatic major depressive disorder (MDD). Using the unique opportunity afforded by the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) study data, we examined whether DMV with afternoon or evening worsening, in addition to classic early morning worsening, was related to other symptom constructs to determine whether the exclusive reliance on morning worsening is justified in defining melancholic features. METHOD: Baseline demographic and clinical characteristics, as well as depressive symptoms, including DMV, were evaluated in 3744 outpatients with nonpsychotic MDD enrolled in the STAR*D study. RESULTS: DMV in at least one of the time periods was reported by 22.4% (N = 837) of the sample. Only 3.3% (N = 28) of these 837 patients with DMV attributed it to environmental factors. Of the 809 participants (96.7%) with DMV unrelated to environmental events, only 31.9% (N = 258) reported morning worsening, while 19.5% (N = 158) and 48.6% (N = 393) reported afternoon and evening worsening, respectively. Minimal distinctions in demographic characteristics, clinical features, and depressive symptoms were found between participants with morning worsening and those with either afternoon or evening worsening. More importantly, other melancholic symptom features were associated with DMV regardless of time of worsening. CONCLUSION: DMV was meaningfully related to other melancholia criteria regardless of when the DMV occurred. If replicated, these findings suggest that DMV as a component of melancholic features might be expanded to include any DMV, not simply early morning worsening. PMID- 17915972 TI - Family intervention approach to loss of clinical effect during long-term antidepressant treatment: a pilot study. AB - BACKGROUND: The return of depressive symptoms during maintenance antidepressant treatment is a common phenomenon, but has attracted very limited research attention. The aims of this investigation were to explore the feasibility of a family intervention approach to loss of clinical effect during long-term antidepressant therapy and to compare this approach with dose increase. METHOD: Twenty outpatients with recurrent major depressive disorder (diagnosed using Research Diagnostic Criteria, i.e., patients were at their third or greater episode of major depressive disorder, with the immediately preceding episode being no more than 2.5 years before the onset of the episode which led to antidepressant treatment) who lived with a partner and relapsed while taking antidepressant drugs were randomly assigned to (1) family intervention approach according to the McMaster Model and maintenance of the anti-depressant drug at the same dosage or (2) dose increase and clinical management. A 1-year follow-up was performed. The study was conducted from January 2002 to December 2004. RESULTS: Seven of 10 patients responded to an increased dosage; all but 1 relapsed again on that dosage during follow-up. Seven of 10 patients responded to family intervention, but only 1 relapsed during follow-up. The difference in relapse was significant (p < .05). CONCLUSIONS: The data suggest that application of a family intervention approach is feasible when there is a loss of clinical effect during long-term antidepressant treatment, and this approach may carry long-term benefits. The results need to be confirmed by large-scale controlled studies but should alert the physician to explore the psychosocial correlates of loss of clinical effect. PMID- 17915973 TI - Substance use disorders and overweight/obesity in bipolar I disorder: preliminary evidence for competing addictions. AB - OBJECTIVE: This investigation was undertaken to explore the relationship between alcohol/illicit drug dependence and overweight/obesity in individuals with bipolar I disorder. METHOD: The data for this analysis were procured from the Canadian Community Health Survey-Mental Health and Well-Being (CCHS) conducted by Statistics Canada in 2002. Bipolar I disorder was defined as persons screening positive for a lifetime manic episode using the World Mental Health 2000 version of the Composite International Diagnostic Interview (WMH-CIDI). Substance abuse and illicit drug dependence were determined using criteria commensurate with the DSM-IV-TR. Overweight and obesity were defined as a body mass index of 25.0 to 29.9 and greater than or equal to 30.0 kg/m(2), respectively. RESULTS: The total sample comprised 36,984 individuals (>or= 15 years old) screening positive for a lifetime manic episode. Subgroup analysis indicated that overweight/obese bipolar individuals had a significantly lower rate of substance dependence than the normal weight sample (13% vs. 21%, p < .01). Conversely, bipolar individuals who screened positive for substance dependence had a lower rate of overweight/obesity when compared with non-substance-dependent bipolar respondents (39% vs. 54%, p< .01). The inverse association between the presence of these 2 co-morbid conditions in bipolar I disorder continued to be statistically significant in multivariate analysis (OR = 0.57, 95% CI = 0.34 to 0.95, p < .05). CONCLUSION: An inverse relationship between the presence of comorbid overweight/obesity and substance use disorders was observed in bipolar I disorder. These results suggest that comorbid addictive disorders (i.e., substance use and compulsive overeating) may compete for the same brain reward systems. PMID- 17915974 TI - Association of AKT1 gene polymorphisms with risk of schizophrenia and with response to antipsychotics in the Chinese population. AB - BACKGROUND: A number of studies have pointed to the involvement of AKT signaling pathways in the etiology of schizophrenia. The purpose of this study was to determine whether the AKT1 gene is involved in the etiology of schizophrenia and whether it affects therapeutic outcomes in the Chinese population. METHOD: Five single nucleotide polymorphisms (SNPs) were genotyped among 384 schizophrenic patients (DSM-IV criteria) and 384 healthy controls from the Chinese population. We systematically analyzed the association of the AKT1 gene with schizophrenia on the basis of sex, age at onset, therapeutic response to typical antipsychotics and atypical antipsychotics, and presence or absence of extrapyramidal syndrome. The study was conducted from May 2004 to June 2006. RESULTS: We found a positive association of the G allele of the SNP marker rs3803300 with schizophrenia (p = .003), both in early-onset and late-onset subjects, and that a haplotype A-G-C-G A constructed by the 5 SNPs showed significant association (p = .00004886). However, we found no relationship between any of the 5 SNP markers and therapeutic response to typical and atypical antipsychotics and chlorpromazine induced extrapyramidal syndrome. CONCLUSIONS: Our study suggests that AKT1 is a susceptibility gene for schizophrenia in the Chinese population and that the AKT1 gene may play no major role in the therapeutic response to antipsychotics or in chlorpromazine-induced extrapyramidal syndrome. PMID- 17915975 TI - A 24-month prospective outcome study of first-episode schizophrenia and schizoaffective disorder within an early psychosis intervention program. AB - BACKGROUND: Within the schizophrenia spectrum disorders, schizoaffective disorder is conventionally considered to have a better prognosis compared to schizophrenia. We tested the hypothesis that patients with first-episode schizoaffective disorder had better clinical and functional outcomes compared to patients with first-episode schizophrenia. METHOD: The study population consisted of consecutive subjects with first-episode schizophrenia or first-episode schizoaffective disorder (diagnosed according to DSM-IV criteria) enrolled in the national Early Psychosis Intervention Program at the Institute of Mental Health/Woodbridge Hospital in Singapore from March 2001 to March 2003. The subjects' level of psychopathology, insight, socio-occupational functioning, and quality of life were assessed using the Positive and Negative Syndrome Scale (PANSS), the Scale to Assess Unawareness of Mental Disorder, the Global Assessment of Functioning scale, and the World Health Organization Quality of Life-BREF scale, respectively, at baseline and at 6, 12, 18, and 24 months. RESULTS: At baseline, patients with schizo-affective disorder (N = 24) were more likely to be employed (OR = 3.38, 95% CI = 1.27 to 9.02, p < .05), had a shorter duration of untreated psychosis (z = -3.30, p < .005), but had greater general psychopathology subscale scores on PANSS (z = -2.69, p < .01) compared to patients with schizophrenia (N = 254). Patients with schizophrenia had better insight into their psychiatric illness (z = -3.93, p < .001) at 6 months and expressed a better level of quality of life in the psychological health domain (z = -3.83, p < .001) at 12 months compared to patients with schizoaffective disorder. At 18 months, patients with schizoaffective disorder continued to have higher general psychopathology subscale scores on PANSS (z = -3.89, p < .001) compared to patients with schizophrenia. CONCLUSION: Patients with first-episode schizoaffective disorder do not necessarily have less severe psychopathology or better longitudinal outcomes compared to patients with first-episode schizophrenia, a finding that warrants attention in the clinical management of these patients. PMID- 17915976 TI - Antimanic response to aripiprazole in bipolar I disorder patients is independent of the agitation level at baseline. AB - OBJECTIVE: To examine the antimanic efficacy of the relatively nonsedating antipsychotic aripiprazole in patients with bipolar I disorder and high or low baseline levels of agitation. METHOD: Data were pooled for this post hoc analysis from two 3-week, placebo-controlled trials of aripiprazole in acute mania (DSM IV). Patients randomly assigned to aripiprazole 30 mg/day (N = 259) or placebo (N = 254) were classified as having either high (Positive and Negative Syndrome Scale [PANSS] Excited Component [PEC] score of >or=14 and a score of >or= 4 on at least one PEC item) or low (PEC < 14) levels of agitation at baseline. Efficacy measures were changes in Young Mania Rating Scale (YMRS) scores, Clinical Global Impressions-Bipolar (CGI-BP) scores, and PEC scores. Efficacy and agitation measurements were assessed by analysis of covariance. RESULTS: From the first week of therapy onward, aripiprazole-treated subjects had significantly greater reduction from baseline in YMRS total scores than placebo-treated subjects in both the high- and low-agitation groups (p < .05 for both groups) and significantly improved CGI-BP scores vs. placebo at end point (p < .05 for both). In highly agitated patients receiving aripiprazole, PEC scores were significantly decreased versus placebo at end point (p < .05). In patients with low agitation receiving aripiprazole, no increases in PEC scores were seen, and a significant reduction in agitation symptoms was apparent after adjustment for baseline PEC scores. CONCLUSIONS: Aripiprazole was superior to placebo in reducing the severity of both mania and agitation in highly agitated patients with bipolar I disorder and showed significant antimanic activity in patients with low levels of agitation without increasing agitation. These findings suggest that aripiprazole's antimanic effect is specific and not limited to control of agitation through sedation. PMID- 17915977 TI - Lamotrigine treatment of pathologic skin picking: an open-label study. AB - BACKGROUND: Although pathologic skin picking is a relatively common behavior, treatment data are limited. We hypothesized that lamotrigine would reduce the symptoms of pathologic skin picking. METHOD: 24 subjects (19 women [79.2%]; mean +/-SD age = 34.1 +/- 12.2 years) with pathologic skin picking (based on DSM-IV criteria for other impulse control disorders) were treated in a 12-week open label trial of lamotrigine as mono-therapy. Lamotrigine dosing ranged from 25 mg every other day to 300 mg/day. The primary outcome measure was time per day spent picking. Subjects were also assessed with measures examining the symptoms of pathologic skin picking and psychosocial functioning. Data were collected from January 15, 2006, to September 18, 2006. RESULTS: Mean (SD) time per day spent picking decreased from 118.1 (130.0) to 59.9 (115.2) minutes (p < .001). Sixteen subjects (66.7%) were considered either "very much improved" or "much improved" in terms of skin picking symptoms. Seven subjects (29.2%) reported no picking at study endpoint. Significant improvement was seen on scales assessing the symptoms of pathologic skin picking (p = .001) and social functioning (p = .002). Mean time to response (i.e., when the subject was much or very much improved) was 8 weeks, which corresponded to a lamotrigine dose of 200 mg/day. CONCLUSIONS: Lamotrigine was associated with improvements in two thirds of subjects with pathologic skin picking. Placebo-controlled, double-blind studies are needed to evaluate further the safety, tolerability, and efficacy of lamotrigine in the treatment of this problematic behavior. TRIAL REGISTRATION: ClinicalTrials.gov identifier NCT00269594 (http://www.clinicaltrials.gov). PMID- 17915978 TI - Depression and risk of nursing home admission among older adults in home care in Europe: results from the Aged in Home Care (AdHOC) study. AB - OBJECTIVE: Depression is a frequent condition observed among nursing home residents. However, so far, limited data are available on the impact of depression on nursing home admission. The aim of the present study was to assess the effect of depression on the risk of nursing home admission in a group of older adults receiving home care in Europe. METHOD: We conducted a longitudinal analysis using data from the Aged in Home Care (AdHOC) database, which contains information on older adults receiving home care services in 11 European countries. Subjects had been admitted to the home care programs between 2001 and 2003. Depression was diagnosed as a score >or=3 on the Minimum Data Set (MDS) Depression Rating Scale. Information on nursing home admission was collected semiannually for 1 year by trained research personnel. RESULTS: The mean age of 2718 older adults entering the study was 82.4 (SD = 7.3) years, 2047 (75.3%) were women, and 331 (12.2%) were depressed. Overall, 49/331 depressed participants (14.8%) and 252/2387 nondepressed participants (10.6%) were admitted to a nursing home (p = .02). After adjusting for potential confounders, the risk of nursing home admission was significantly higher for depressed participants (hazard ratio = 1.43, 95% CI = 1.02 to 2.02). The risk of nursing home admission progressively and significantly increased as MDS Depression Rating Scale score increased (signifying more severe depression) (p = .001 for linear trend). CONCLUSIONS: In older adults receiving home care in Europe, depression is associated with an increased risk for nursing home admission. This association increases with severity of depression. PMID- 17915980 TI - Smoking cessation in patients with psychiatric disorders. PMID- 17915979 TI - Antidepressants in adult suicides in New York City: 2001-2004. AB - BACKGROUND: The U.S. Food and Drug Administration recently extended the Black Box warning on anti-depressants regarding pediatric suicidality to include young adults. The decision was guided by results from meta-analyses of 372 randomized controlled clinical trials of antidepressants for adults. Nearly all suicidality in those trials was nonfatal suicide attempts and ideation. Here, we consider whether antidepressants are linked with adult suicide deaths. METHOD: Subjects in this medical examiner surveillance study included all suicides, 18 years and older, in New York City from 2001-2004. Postmortem blood was analyzed for the presence of antidepressants. RESULTS: There were 1419 adult suicides in New York City during the study period. Antidepressants were detected at autopsy in 23.1% of the suicides who met criteria for toxicology analyses. Antidepressants were least prevalent in suicides aged 18-24 years (13.9%). CONCLUSIONS: Antidepressants were detected in less than one-quarter of adult suicides in New York City from 2001-2004. The majority of the suicides were not attributable to antidepressant use, and perhaps many could have been prevented with appropriate treatment. Although this study does not provide evidence for a link between antidepressant use and subsequent suicide, careful monitoring of patients receiving antidepressants remains critically important. PMID- 17915982 TI - The phenomenology and assessment of childhood and adolescent mental health: progress in research. PMID- 17915981 TI - Phenomenology and prognostic significance of delusions in major depressive disorder: a 10-year prospective follow-up study. AB - OBJECTIVE: The study explored the phenomenology and prognostic significance of delusions in major depressive disorder. METHOD: From 452 patients with DSM-III major depression, we selected those with at least one belief fulfilling both DSM III prerequisites for a delusion (i.e., being of "delusional proportions" and being maintained with "delusional intensity"). These patients were compared to the others with respect to demographic, historical, and index episode features; time spent in a depressive episode during a prospective observation period; and 10-year outcome. The same comparisons were made between patients with mood incongruent delusions and those with mood-congruent delusions only. The study covered the period between January 1, 1978, and December 31, 2005. RESULTS: About 20% of patients had at least one delusion in their index episode. An additional 5.3% had a belief fulfilling only one of the DSM-III prerequisites for a delusion. In about one quarter of delusional patients, the index episode was not "severe." Almost 10% of delusional patients had both mood-congruent and mood incongruent delusions. In patients with delusions, time to syndromal recovery from index episode was longer and antipsychotic medication was more frequently used (both p < .0001). The presence of delusions predicted a higher depressive morbidity during the prospective observation period (p < .05), but not a poorer 10-year outcome. No variable discriminated patients with mood-incongruent delusions from those with mood-congruent delusions only. CONCLUSION: The presence of delusions in a major depressive episode has significant therapeutic and short term prognostic implications. However, the boundary between delusions and nondelusional sustained preoccupations is somewhat fuzzy, and some DSM-IV assumptions concerning psychotic depression (i.e., that this depression is always "severe"; that in an individual patient, delusions will be either all congruent or all incongruent with depressed mood; and that mood-incongruent delusions are associated with a poorer prognosis) may be unwarranted. PMID- 17915983 TI - Association of internalizing disorders and allergies in a child and adolescent psychiatry clinical sample. AB - OBJECTIVE: To investigate the specificity of the association between internalizing disorders (anxiety and depression) and atopic disorders (asthma, allergic rhinitis, urticaria, and atopic dermatitis) in a child and adolescent psychiatric clinical sample. METHOD: A sample of 184 youths was evaluated for current DSM-IV psychiatric disorders (clinical interview) and lifetime history of atopic disorders (parent report and chart review) in a child and adolescent psychiatry clinic from September 1, 2001, through December 31, 2002. Logistic regression analyses were used to assess the differential likelihood of having a lifetime history of atopic disorders among psychiatrically ill youths with and without internalizing disorders. RESULTS: Youths with internalizing disorders were significantly more likely than those with noninternalizing disorders to have a lifetime history of atopic disorders (odds ratio [OR] = 1.95, 95% CI = 1.02 to 3.73, p = .04). Moreover, analyses distinguishing youths with "pure" internalizing disorders from those with comorbid internalizing and externalizing disorders, "pure" externalizing disorders, and other psychiatric disorders showed that the association with atopic disorders was specific for "pure" internalizing disorders only (OR = 2.40, 95% CI = 1.09 to 5.30, p = .03). CONCLUSIONS: Atopic disorders may be associated specifically with "pure" internalizing disorders in psychiatrically ill youths. Additional studies are needed to identify the underlying mechanisms of this specificity for the subsequent development of effective treatment and prevention interventions that target both disorders. PMID- 17915984 TI - Association between mental health status and sleep status among adolescents in Japan: a nationwide cross-sectional survey. AB - OBJECTIVE: Previous epidemiological studies on relationships between mental health status and sleep status of adolescents have not been sufficiently representative. In the present study, using samples representative of Japanese adolescents nationwide, associations between mental health status and various sleep statuses were examined. METHOD: The survey was conducted in December 2004 and January 2005 among students enrolled in randomly selected junior and senior high schools throughout Japan, using self-administered questionnaires that addressed lifestyle, sleep status, mental health status, and personal data. Of 103,650 questionnaires collected, 99,668 were analyzed. Sleep status was assessed according to sleep duration, subjective sleep assessment, bedtime, and insomnia symptoms. The Japanese version of the 12-item General Health Questionnaire was employed for assessment of mental health status. RESULTS: Mental health status of subjects whose sleep duration was less than 7 hours, and those who slept 9 hours or more, was poorer than that of subjects who slept for 7 hours or more but less than 9 hours. A U-shaped association was observed between mental health status and sleep duration. Furthermore, a linear association was observed between subjective sleep assessment and mental health status; the worse the subjective sleep assessment, the poorer the mental health status. Mental health status was also inversely proportional to the frequency of insomnia symptoms. CONCLUSION: The fact that sleep duration and subjective sleep assessment showed different patterns of association with mental health status indicates that these 2 sleep parameters have independent significance. Considering these associations, it is important to promote mental health care and sleep hygiene education for adolescents. PMID- 17915985 TI - A pilot study of an electronic, adolescent version of the quick inventory of depressive symptomatology. AB - BACKGROUND: Adolescent depression assessments are time-intensive, often requiring separate interviews with an adolescent and a parent/ informant. In adults, a self rated, interactive voice response (IVR) version of the Quick Inventory of Depressive Symptomatology (QIDS-IVR) has been shown to be reliable, valid, and sensitive to change. An adolescent version of the QIDS (QIDS-A-IVR) was created using speaker-independent voice recognition technology. An informant version, QIDS-P-IVR, collects ratings from parents or other knowledgeable adults. METHOD: The study included 27 adolescents ranging from 12 to 17 years of age, 48% of whom were female. During a single office visit, adolescents completed the QIDS-A-IVR and parents completed the QIDS-P-IVR. A clinician completed the clinician-rated adult version of the QIDS separately for adolescents (QIDS-C-A) and parents (QIDS C-P) and the Children's Depression Rating Scale-Revised (CDRS-R). The study was conducted from October 2005 to April 2006. RESULTS: Cronbach alpha of the QIDS-A IVR was .85. The QIDS-A-IVR correlated significantly with the QIDS-C-A (r = 0.95) and the CDRS-R (r = 0.76), both p < .01. Conversely, the correlations of the QIDS A-IVR with the QIDS-P-IVR and the QIDS-C-P were small and nonsignificant. The QIDS-A-IVR required adolescents a mean of 6 minutes and 31 seconds to complete (SD = 41 seconds). The voice recognition technology correctly identified the adolescents' spoken words in 92% of the 483 spoken responses. The system recognized a response from all adolescents on all items. CONCLUSIONS: This study supports the reliability and validity of the QIDS-A-IVR as an adolescent depression measure. The QIDS-A-IVR may provide clinicians and researchers with a sound, technology-based method of assessing adolescent depression. Future research is needed on the informational value of parent ratings of adolescent depression. PMID- 17915987 TI - Olanzapine and blepharoclonus. PMID- 17915988 TI - Physiologic sleep disorders among treatment-responsive depressed patients with residual cognitive and physical symptoms. PMID- 17915990 TI - Is smoking associated with suicide in bipolar patients? PMID- 17915989 TI - Aripiprazole-induced psychosis: a case report of reexposure by stepwise up titration. PMID- 17915993 TI - Computer-assisted evaluation of kinematics of the two bundles of the anterior cruciate ligament. AB - The aim of this cadaveric study was to describe the kinematics of the anterior cruciate ligament (ACL)-intact, posterolateral (PL) bundle-deficient and ACL deficient knee by applying a protocol for computer-assisted evaluation of knee kinematics. The hypothesis that the PL bundle functions mainly at low knee flexion angles was tested. An optical tracking system was used to acquire knee joint motion on 10 knees during clinical evaluations by tracking markers rigidly attached to the bones. The protocol included acquisition of anterior-posterior (AP) translations and internal-external (IE) rotations, and evaluation of three clinical knee laxity tests (anterior drawer, manual and instrumented Lachman). The data demonstrated no significant contribution to AP translation and IE laxity from the PL bundle over the entire range of motion. The clinical knee laxity tests showed no significant differences between the ACL-intact and PL bundle deficient states. The hypothesis could not be proven. Current clinical knee laxity measurements may not be suited for detecting subtle changes such as PL bundle deficiency in the ACL anatomy. The computation of knee laxity might be a step towards a more precise kinematic test of knee stability not only in the native and torn ACL state of the knee but also in the reconstructed knee. PMID- 17915994 TI - [The relation of workspace and installation space of epicyclic kinematics with six degrees of freedom]. AB - INTRODUCTION: The kinematics of a robotic device significantly determines its installation space when it comes to technical realisation. With regard to the deployment of robotic manipulators in surgery, manipulators with a preferably small installation space are needed. MATERIALS AND METHODS: This study describes six versions of novel epicyclic kinematics with six degrees of freedom (DOF). At first, the kinematics functionality was analysed using Gruebler's formula. Subsequently, the quantitative determination of the relation of workspace and installation space was performed using Matlab algorithms. To qualitatively describe the shape of the workspace, the Matlab visualisation features were utilised. For comparison, the well-known Hexapod was used. RESULTS: The assessed kinematics had 6-DOF-functionality. It became apparent that one version of the epicyclic kinematics having two 3-DOF disk systems mounted in a parallel way featured a particularly good relation of workspace and installation space. Compared to the Hexapod, this is approximately four times better. The shape of the workspaces of all epicyclic kinematics assessed was convex and compact. CONCLUSION: It could be shown that a novel epicyclic kinematics has a notably advantageous relation of workspace and installation space. Apparently, it seems to be well suited for the deployment in robotic machines for surgical procedures. PMID- 17915995 TI - [Biomechanical modeling of the cervical spine on the basis of tomographic data]. AB - Navigation systems have been successfully established in spine surgery over the past years. Although there is rather good intraoperative support, preoperative planning still lacks a technologically mature software basis. In particular, the computation of the effect of intervention would be of great importance to the surgeon, which would allow him to choose the right strategy. For the patient, this would result in reduced trauma and optimized postoperative mobility. For a correct simulation of the behavior of the cervical spine, a patient individual finite element model is needed which would precisely reflect the geometrical and biomechanical properties of the spine. Therefore, a modeling process starting from tomographic data and ending in an applicable finite element model must be elaborated. A possible solution will be presented in this paper. PMID- 17915996 TI - [The bending rod prosthesis: an experimental approach to metaphyseal hip arthroplasty]. AB - The aim of our study was to develop a femoral component for total hip arthroplasty that would exclusively anchor in the metaphysis of the femoral neck. To forego trochanteric fixation, the load needs to be transferred to the metaphysis at as many points as possible. A computer simulation model suggested that an implant with a central cylinder and 16 rods aligned along a thread would be the preferable solution. To evaluate primary implantation stability, 14 fresh frozen cadaver femora were used. A special instrument set was developed to allow for centered implantation of the prosthesis without the need to dissect the greater trochanter. For our tests, we used two prototype implants: one made from titanium and the other from a CoCrMo alloy. For the measurement of micromotions at the medial proximal femur, sinusoid dynamic loading with a force between 300 N and 1700 N and a frequency of 1 Hz was employed. In a neutral position of 16 degrees adduction and 9 degrees antetorsion, the average micromotions measured were 119 microm. Despite these convincing in vitro results with regards to primary stability, circular cut-out of the implant, followed by aseptic osteonecrosis, loosening might still occur in a clinical situation. Animal experiments are therefore required to further evaluate this new implant design. PMID- 17915997 TI - [Light microscopic examination of rabbit skulls following conventional and Piezosurgery osteotomy]. AB - INTRODUCTION: The novel ultrasonic osteotomy technique (Piezosurgery) is an alternative to conventional osteotomy devices. The aim of the present study was to carry out morphological comparison of the bone surface using conventional osteotomy techniques in comparison to the rather new ultrasonic osteotomy technique by means of a reflected-light microscopic examination. MATERIALS AND METHODS: Following the sacrifice of 12 rabbits, 24 standardized bone samples were removed from the skull. The osteotomy devices used were a rotating instrument (Lindemann bur), an oscillating micro-saw, and an ultrasonic osteotomy device (Piezosurgery) with insert tips OT6 and OT7. The times needed for osteotomy were measured. The bone surfaces were examined using a reflected-light microscope with a magnification of 40x and 100x. RESULTS: Osteotomy with Piezosurgery is significantly more time consuming than osteotomy with conventional methods (p<0.05). Following osteotomy with the ultrasonic device, the reflected-light microscopic examinations of the unmodified bone samples revealed typical bone structure of the calvaria, including compacta externa, diploe and compacta interna. On the contrary, following osteotomy with the conventional devices, the diploe structure presented distinct modifications. The cancellous spaces were filled with bone debris, and the cancellous structure was demolished. The samples prepared by the micro-saw technique showed a superficially condensed and grooved surface. CONCLUSION: In the present study, well-defined differences were observed following osteotomy with conventional devices and osteotomy with the ultrasonic device. The integrity of the bony structure observed after the ultrasonic technique could benefit the bone healing process. Further studies dealing with the bone healing process after using different osteotomy techniques are recommended. PMID- 17915998 TI - Sudden failure of implantable pulse generators: cause of failure and examination. AB - A sudden failure of implantable pulse generators used for spinal cord stimulation occurred in two patients. To identify the cause of this failure, an intensive destructive analysis of the explanted devices was carried out. A functional diagnosis was carried out by inspecting amplitude, pulse width and frequency on each output channel of the implantable pulse generators. Later, the titanium case of the pulse generators was opened by laser cutting to minimise any additional mechanical stress during the opening procedure. The functional test for both pulse generators showed faultless behaviour. Using light and electron microscopy, hairline cracks could be identified in the electrical connection between battery and electronic circuit. In both devices, the cracks spread through the whole bond wire in the connection to the plus pole of the battery and partially also to the minus pole. The analysis showed that both devices failed by broken bond wires. The electrical connection to the battery exists just by the spring characteristic of the wires. A push to the implant causes a short-term disconnection, resulting in a power on reset of the device. Manufacturing or design issues, allowing micromotion between battery and the hybrid part, may be the reason for this problem. PMID- 17916000 TI - Effects of marrow penetration on bone augmentation within a titanium cap in rabbit calvarium. AB - BACKGROUND: Some studies have advocated marrow penetration in guided bone augmentation (GBA), whereas others have shown that bone can be generated without marrow penetration. This study examined the effect of marrow penetration and the optimal rate of penetration. METHODS: In 10 rabbits, the calvarium was exposed, and circular grooves were prepared bilaterally. Within the circular groove, the external cortical surface of the skull was perforated mechanically using a number 4 round bur (experimental site), whereas the bone surface on the other groove (control site) was left intact. The rate of penetration was standardized as 28% for the experimental site and 0% for the control site. Subsequently, two standardized titanium caps with an inner diameter of 8 mm and an inner height of 4 mm were anchored in the prepared grooves. Five rabbits each were euthanized at 1 and 3 months, and the percent area of newly generated tissue and mineralized bone in the newly generated tissue under the titanium cap was determined. RESULTS: Histomorphometric analysis showed significantly increased bone neogenesis in the experimental site at 3 months. The percent area of mineralized bone in the newly generated tissue was consistently higher in the experimental site at 3 months. CONCLUSION: In the rabbit calvarium using a titanium cap GBA model, bone augmentation was significantly greater with marrow penetration than without penetration. PMID- 17915999 TI - Effect of adjunctive systemic azithromycin with periodontal surgery in the treatment of chronic periodontitis in smokers: a pilot study. AB - BACKGROUND: Along with conventional surgical therapy, systemic antibiotics may provide more effective treatment in smokers by targeting tissue-invasive bacteria. The aim of this randomized, placebo-controlled, double-masked clinical trial was to evaluate the adjunctive effects of systemic azithromycin (AZM) in combination with periodontal pocket reduction surgery in the treatment of chronic periodontitis in smokers. METHODS: Thirty patients with a greater than one pack/day smoking habit and generalized moderate to severe chronic periodontitis were randomized to the test (surgery plus 3 days of AZM, 500 mg) or control group (surgery plus 3 days of placebo). Full-mouth probing depth (PD), clinical attachment level (CAL), bleeding on probing (BOP), gingival index (GI), plaque index, and wound healing indices (WHI) were assessed at baseline and at 2 weeks and 1, 3, and 6 months following surgical intervention. Plaque and gingival crevicular fluid were collected for trypsin-like enzyme activity (benzoyl-dl arginine naphthylamine) and bone biomarker (crosslinked telopeptide of type I collagen [ICTP]) analyses, respectively, at baseline, 2 weeks, and 1, 3, and 6 months. RESULTS: Surgical treatment of moderate (PD = 4 to 6 mm) and deep (PD > 6 mm) pockets significantly improved clinical parameters of treated and untreated teeth (CAL gain, PD reduction, and reduction of BOP). The additional use of AZM did not enhance this improvement nor did it promote reduction of ICTP levels. Compared to the control group, the test group had significantly better WHI scores at 1 month, significantly less GI at 2 weeks, and sustained reductions of red complex bacteria with trypsin-like enzyme activity at 3 months. For non-surgery teeth, only the test group showed significant gains in overall CAL compared to baseline. CONCLUSIONS: The findings of this pilot study demonstrated that in heavy smokers, adjunctive systemic AZM in combination with pocket reduction surgery did not significantly enhance PD reduction or CAL gain. However, the clinical value of adjunctive AZM may be appreciated by more rapid wound healing, less short-term gingival inflammation, and sustained reductions of periopathogenic bacteria. More expanded studies are recommended to better determine the clinical effects of adjunctive AZM in patients who smoke. PMID- 17916001 TI - Chronic administration of lipopolysaccharide and proteases induces periodontal inflammation and hepatic steatosis in rats. AB - BACKGROUND: Epidemiologic studies suggest a relationship between periodontitis and liver diseases. A rat periodontitis model was used to investigate whether a causal relationship exists between periodontitis and liver diseases. METHODS: Fourteen male Wistar rats (8 weeks old) were divided into two groups: a periodontitis group in which Escherichia coli lipopolysaccharide (LPS) and Streptomyces griseus proteases were applied into the gingival sulcus for 8 weeks, and a control group using pyrogen-free water instead. After blood samples were collected, periodontal tissues and liver specimens were analyzed. RESULTS: Chronic administration of LPS and proteases to the gingival sulcus induced periodontitis and liver injury, including steatosis with inflammation and sinusoidal fibrosis. Apoptosis, enhanced concentration of 8 hydroxydeoxyguanosine, and activated production of tumor necrosis factor-alpha in liver were observed in the periodontitis group, with increased gingival inflammation, serum LPS, and reactive oxygen species. CONCLUSION: Periodontal inflammation in a rat model induced fatty liver disease through increased serum LPS. PMID- 17916002 TI - Thank you, Herb Gilman. PMID- 17916007 TI - Pharmacokinetics of gallium maltolate after intragastric administration in neonatal foals. AB - OBJECTIVE: To determine the pharmacokinetics of gallium maltolate (GaM) after intragastric administration in healthy foals. ANIMALS: 6 healthy neonatal foals. PROCEDURES: Each foal received GaM (20 mg/kg) by intragastric administration. Blood samples were obtained before (time 0) and at 0.25, 0.5, 1, 2, 4, 8, 12, 24, 36, and 48 hours after GaM administration for determination of serum gallium concentrations by use of inductively coupled plasma mass spectroscopy. RESULTS: Mean +/- SD pharmacokinetic variables were as follows: peak serum gallium concentration, 1,079 +/- 311 ng/mL; time to peak serum concentration, 4.3 +/- 2.0 hours; area under the serum concentration versus time curve, 40,215 +/- 8,420 ng/mL/h; mean residence time, 39.5 +/- 17.2 hours; area under the moment curve, 1,636,554 +/- 931,458 ng([h](2)/mL); and terminal half-life, 26.6 +/- 11.6 hours. The mean serum concentration of gallium at 12 hours was 756 +/- 195 ng/mL. CONCLUSIONS AND CLINICAL RELEVANCE: Gallium maltolate administered via nasogastric tube at a dose of 20 mg/kg to neonatal foals resulted in gallium serum concentrations considered sufficient to suppress growth or kill Rhodococcus equi in macrophages and other infected tissues. PMID- 17916008 TI - Modulation of the tissue reninangiotensin-aldosterone system in dogs with chronic mild regurgitation through the mitral valve. AB - OBJECTIVE: To investigate whether the tissue and plasma renin-angiotensin aldosterone system (RAAS) is activated in dogs with mild regurgitation through the mitral valve and determine the contribution of chymase and angiotensin converting enzyme (ACE) to the activation of the RAAS and potential production of angiotensin II during the chronic stage of mild mitral valve regurgitation. ANIMALS: 5 Beagles with experimentally induced mild mitral valve regurgitation and 6 clinically normal (control) Beagles. PROCEDURES: Tissue ACE and chymase like activities and plasma RAAS were measured and the RAAS evaluated approximately 1,000 days after experimental induction of mitral valve regurgitation in the 5 dogs. RESULTS: Dogs with experimentally induced mitral valve regurgitation did not have clinical signs of the condition, although echocardiography revealed substantial eccentric hyper- trophy. On the basis of these findings, dogs with mitral valve regurgitation were classified as International Small Animal Cardiac Health Council class Ib. Plasma activity of renin and plasma concentrations of angiotensin I, angiotensin II, and aldosterone were not significantly different between dogs with mitral valve regurgitation and clinically normal dogs. Tissue ACE activity was significantly increased and chymase-like activity significantly decreased in dogs with mitral valve regurgitation, compared with values in clinically normal dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The tissue RAAS was modulated without changes in the plasma RAAS in dogs with mild mitral valve regurgitation during the chronic stage of the condition. An ACE-dependent pathway may be a major route for production of angiotensin II during this stage of the condition. PMID- 17916009 TI - Assessment of longitudinal tissue Doppler imaging of the left ventricular septum and free wall as an indicator of left ventricular systolic function in dogs. AB - OBJECTIVE: To evaluate tissue Doppler imaging (TDI) of the left ventricular (LV) free wall (FW) and ventricular septum (VS) as an indicator of LV systolic function in dogs. ANIMALS: 7 healthy Beagles. PROCEDURES: Doses of dobutamine (5 and 10 microg/kg/min) and esmolol (50 and 100 microg/kg/min) were infused into the LV of each dog. With each dose, heart rate; myocardial performance index (MPI); transmitral inflow and ejection time (determined via pulsed-wave Doppler [PWD] echocardiography); and FW and VS velocities of the mitral valve annulus (determined via TDI during systole [S'], early diastole [E'], and late diastole [A']) were assessed. RESULTS: With each dose, dobutamine significantly increased heart rate and the first derivatives of LV pressure (+dP/dt and -dP/dt), whereas esmolol significantly decreased the +dP/dt and -dP/dt values, compared with baseline. Esmolol (100 microg/kg/min) significantly decreased the VS-TDI-derived S' velocity and FW-TDI-derived E' velocity; dobutamine significantly increased transmitral inflow and TDI velocities. Regression coefficient between VS-TDI derived S' velocity and +dP/dt was higher than that between FW-TDI-derived S' velocity and +dP/dt. Compared with baseline, the PWD- and VS-TDI-derived MPI were significantly decreased by dobutamine and significantly increased by esmolol at each dose. Values of FW-TDI-derived MPI were higher than values derived via the other techniques. Correlation between +dP/dt and VS-TDI-derived MPI was greater than that between +dP/dt and FW-TDI- or PWD-derived MPI. CONCLUSIONS AND CLINICAL RELEVANCE: In healthy dogs, the VS-TDI-derived S' velocity and MPI appear to be reliable assessments for evaluating LV systolic function. PMID- 17916010 TI - Effects of oral administration of furosemide and torsemide in healthy dogs. AB - OBJECTIVE: To investigate the diuretic effects, tolerability, and adverse effects of furosemide and torsemide after short- and long-term administration in healthy dogs. ANIMALS: 8 mixed-breed dogs. PROCEDURES: In a crossover study, furosemide (2 mg/kg), torsemide (0.2 mg/kg), or placebo (bifidobacterium [1 mg/kg]) was administered orally to each dog every 12 hours for 14 days. Blood and urine samples were collected before the study (baseline data) and at intervals on the 1st (short-term administration) and 14th day (long-term administration) of treatment for assessment of urine volume and specific gravity and selected clinicopathologic variables including BUN, creatinine, and aldosterone concentrations, and creatinine clearance. RESULTS: Compared with the baseline value, short-term administration of furosemide or torsemide immediately increased urine volume significantly; after long-term administration of either drug, urine specific gravity decreased significantly. Compared with the effect of placebo, the 24-hour urine volume was significantly increased after short-term administra tion of furosemide or torsemide. In addition, it was significantly increased after long-term administration of torsemide, compared with that of short-term administration. Long-term administration of furosemide or torsemide increased the BUN and plasma creatinine con-centrations, compared with the baseline value. Compared with the baseline value, plasma aldosterone concentration was significantly increased after long-term administration of either drug and was significantly higher after torsemide treatment than after furosemide treatment. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, diuretic resistance developed after 14 days of furosemide, but not torsemide, administration; however, both loop diuretics were associated with increased BUN and plasma creatinine concentrations, compared with values before treatment. PMID- 17916011 TI - Evaluation of thermographic imaging of the limbs of healthy dogs. AB - OBJECTIVE: To describe a thermographic imaging protocol, identify normal thermographic patterns (ie, color map reflecting the skin temperature distribution) for various regions of interest (ROIs) of dog limbs, and evaluate effects of clipping the coat on thermographic patterns and limb temperature in healthy dogs. ANIMALS: 10 healthy dogs. PROCEDURES: Each dog was thermographically evaluated in the same room (ambient temperature, 21 degrees C) via ROIs that included cranial and caudal views of the body, full lateral body views, full views of the limbs, and views of various limb regions. After initial imaging, the coat was clipped on the forelimbs and hind limbs only. Each dog was then evaluated 15 and 60 minutes and 24 hours after clipping by use of the same protocol. RESULTS: For each ROI within a category (intact coat and each time point after clipping), mean temperatures were similar among the 10 dogs. Pairwise comparisons for 15 and 60 minutes and 24 hours established patterns of temperature stabilization among the 3 time points. Temperatures did not differ significantly between the left and right limbs. There was a mean success rate of 75% for use of image pattern analysis for recognition of similar thermographic patterns in the forelimbs and hind limbs. CONCLUSIONS AND CLINICAL RELEVANCE: Thermography can be a viable, noninvasive imaging modality that provides consistent images with reproducible thermal patterns in ROIs examined in healthy dogs. Although the coat had a predictable influence to decrease the mean temperature, thermal patterns remained fairly consistent after the coat was clipped. PMID- 17916012 TI - Digestive enzyme concentrations and activities in healthy pancreatic tissue of horses. AB - OBJECTIVE: To measure concentrations and activities of major digestive enzymes in healthy equine pancreatic tissue. ANIMALS: 7 adult horses with normal pancreatic tissues. PROCEDURES: Small pieces of pancreatic tissue were collected immediately after euthanasia, immersed in liquid nitrogen, and maintained at -80 degrees C until analyzed. Concentrations and activities of amylase, lipase, chymotrypsin, trypsin, and elastase were determined by use of a microtiter technique. Relative pancreatic protein concentrations were determined by use of bovine serum albumin as the standard. Pancreatic DNA was extracted and con-centrations determined by use of the diphenylamine method with calf thymus DNA as the standard. RESULTS: The pancreatic cellular concentration of each enzyme, expressed as units per milligram of DNA, was consistent among horses. Cellular concentration of lipase (1,090.8 +/- 285.3 U/mg of DNA) was highest, followed by amylase (59.5 +/- 9.8 U/mg of DNA). Elastase, trypsin, and chymotrypsin were detected in small concentrations (1.9 +/- 0.6, 3.5 +/- 1.5, and 9.6 +/- 2.9 U/mg of DNA, respectively). Similar results were obtained for specific activities of the enzymes. CONCLUSIONS AND CLINICAL RELEVANCE: Results were unexpected because, under natural conditions, the predominant energy source for horses is carbohydrate. These results may indicate, in part, the reason horses seem to tolerate large amounts of fat added to their diet. PMID- 17916013 TI - Associations between genotypes at codon 171 and 136 of the prion protein gene and production traits in market lambs. AB - OBJECTIVE: To determine whether selection for the homozygous A136 R171 genotype that confers resistance to classic scrapie infection negatively affects production traits in sheep. ANIMALS: 996 commercial lambs obtained from 2 flocks at separate locations across 3 consecutive years. Procedures-Genotyping at codon 136 and 171 was performed by use of commercially available testing or a single nucleotide polymorphism assay. Carcass data were collected without knowledge of genotype approximately 24 hours after slaughter by an experienced grader. The model to analyze associations between prion protein (PRNP) genotype and production traits was based on genotype, breed, or both as fixed effects and days on feed as a covariate. RESULTS: Average daily gain was significantly associated with only combined codons 136 and 171. In flock 1, weaning average daily gain was significantly greater in AA136 sheep than heterozygotes; the difference between QR171 and RR171 sheep, compared with QQ171 sheep, were not significant although QR171 and RR171 sheep had higher values. However, in flock 2, average daily gain was significantly greater in AV136 sheep than AA136 sheep and in QR171 sheep than QQ171 sheep. CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggest there is an advantage for average daily gain in lambs with an arginine allele at codon 171, but there were no other genotype effects on production traits. Thus, selection for the resistant arginine allele at codon 171 to comply with USDA scrapie eradication guidelines should not be detrimental to lamb production in commercial flocks. Effects of codon 136 on average daily gain were ambiguous. PMID- 17916014 TI - Biochemical and genetic evaluation of the role of AMP-activated protein kinase in polysaccharide storage myopathy in Quarter Horses. AB - OBJECTIVE: To evaluate whether biochemical or genetic alterations in AMP activated protein kinase (AMPK) play a role in the development of polysaccharide storage myopathy (PSSM) in Quarter Horses. ANIMALS: 30 PSSM-affected and 30 unaffected (control) Quarter Horses. PROCEDURES: By use of an established peptide phosphotransfer assay, basal and maximal AMPK activities were measured in muscle biopsy samples obtained from 6 PSSM-affected and 6 control horses. In 24 PSSM affected and 24 control horses, microsatellite markers identified from the chromosomal locations of all 7 AMPK subunit genes were genotyped with a fluorescent DNA fragment analyzer. Alleles of 2 of the AMPK gamma subunit genes were genotyped via DNA sequencing. Allele frequencies of DNA markers in or near the AMPK subunit genes were measured in isolated genomic DNA. RESULTS: No differences in basal or maximal muscle AMPK enzyme activities between PSSM affected and control horses were detected. There were also no differences in allele frequencies for microsatellite markers near any of the 7 AMPK subunit genes between the 2 groups. Furthermore, previously known and newly identified alleles of 2 equine AMPK gamma subunit genes were also not associated with PSSM. CONCLUSIONS AND CLINICAL RELEVANCE: These results have provided no evidence to indicate that AMPK plays a causative role in PSSM in American Quarter Horses. PMID- 17916015 TI - Efficacy of tinidazole for treatment of cats experimentally infected with Tritrichomonas foetus. AB - OBJECTIVE: To determine the efficacy of tinidazole for treatment of cats with experimentally induced Tritrichomonas foetus infection. ANIMALS: 8 specific pathogen-free kittens. PROCEDURES: Tinidazole was tested for activity against a feline isolate of T foetus in vitro. Kittens were infected orogastrically with the same isolate and treated or not with tinidazole (30 mg/kg, PO, q 24 h for 14 days). Amoxicillin was administered 28 weeks after completion of tinidazole administration to induce diarrhea. Feces were repeatedly tested for T foetus by use of PCR assay and microbial culture for 33 weeks. RESULTS: Tinidazole killed T foetus at concentrations >or= 10 microg/mL in vitro. In experimentally induced infection, tinidazole administered at 30 mg/kg decreased T foetus below the limit of molecular detection in 2 of 4 cats. Recrudescent shedding of T foetus, as elicited by amoxicillin-induced diarrhea, was diminished in cats that received prior treatment with tinidazole. CONCLUSIONS AND CLINICAL RELEVANCE: Although tinidazole decreased the detection of T foetus and treated cats were resistant to later efforts to incite the infection, inability of tinidazole to eradicate infection in many cats poses a serious impediment to the drug's effectiveness in practice. PMID- 17916016 TI - Accuracy of asymmetry indices of ground reaction forces for diagnosis of hind limb lameness in dogs. AB - OBJECTIVE: To determine the accuracy of asymmetry indices of ground reaction forces (GRF) for diagnosis of hind limb lameness in dogs. ANIMALS: 36 healthy dogs and 13 dogs with naturally acquired cranial cruciate ligament rupture or hip dysplasia. PROCEDURES: Lameness for affected dogs ranged from not detectable to minor and constant. While dogs trotted on an instrumented treadmill, GRF variables were recorded and analyzed with asymmetry indices. Each index was tested for its ability to discriminate between healthy and affected dogs. Combinations of several indices were also assessed. RESULTS: Vertical force variables had better accuracy than craniocaudal force variables. Peak vertical force was the most accurate variable. Partial asymmetry during trotting was detected in healthy dogs. A multivariate approach that used peak vertical force and maximal rising slope yielded the optimum combination to distinguish between healthy and affected dogs. In addition, sensitivity of 92% or specificity of 95% may be achieved with 2 cutoff values while simultaneously maintaining specificity or sensitivity, respectively, at > 85%. CONCLUSIONS AND CLINICAL RELEVANCE: Asymmetry indices of GRFs were accurate for detection of hind limb lameness in dogs. This is particularly relevant for study designs in which only a single gait evaluation is possible. PMID- 17916018 TI - Assessment of antiepileptic drugs as substrates for canine P-glycoprotein. AB - OBJECTIVE: To determine whether antiepileptic drugs (AEDs) are substrates for canine P-glycoprotein (P-gp). Sample Population-OS2.4/Doxo cells (canine osteosarcoma cells induced via exposure to doxorubicin to highly express P-gp). PROCEDURES: Competitive inhibition of rhodamine 123 efflux from OS2.4/Doxo cells was used to determine whether AEDs were substrates for canine P-gp. Flow cytometry was used to quantify mean fluorescence intensity of cells treated with rhodamine alone and in combination with each experimental drug. RESULTS: Known P gp substrate drugs ivermectin and cyclosporin A altered rhodamine efflux by 90% and 95%, respectively. Experimental drugs altered rhodamine efflux weakly (diazepam, gabapentin, lamotrigine, levetiracetam, and phenobarbital) or not at all (carbamazepine, felbamate, phenytoin, topirimate, and zonisamide). CONCLUSIONS AND CLINICAL RELEVANCE: At clinically relevant doses, it appeared that AEDs were weak substrates (diazepam, gabapentin, lamotrigine, levetiracetam, and phenobarbital) or were not substrates (carbamazepine, felbamate, phenytoin, topirimate, and zonisamide) for canine P-gp. Therefore, it seems unlikely that efficacy of these AEDs is affected by P-gp expression at the blood-brain barrier in dogs. PMID- 17916017 TI - Isolation and characterization of bone marrow-derived equine mesenchymal stem cells. AB - OBJECTIVE: To isolate and characterize bone marrow-derived equine mesenchymal stem cells (MSCs) for possible future therapeutic applications in horses. SAMPLE POPULATION: Equine MSCs were isolated from bone marrow aspirates obtained from the sternum of 30 donor horses. PROCEDURES: Cells were cultured in medium (alpha minimum essential medium) with a fetal calf serum content of 20%. Equine MSC features were analyzed to determine selfrenewing and differentiation capacity. For potential therapeutic applications, the migratory potential of equine MSCs was determined. An adenoviral vector was used to determine the transduction rate of equine MSCs. RESULTS: Equine MSCs can be culture-expanded. Equine MSCs undergo cryopreservation in liquid nitrogen without altering morphologic characteristics. Furthermore, equine MSCs maintain their ability to proliferate and differentiate after thawing. Immunocytochemically, the expression of the stem cell marker CD90 can be detected on equine MSCs. The multilineage differentiation potential of equine MSCs was revealed by their ability to undergo adipogenic, osteogenic, and chondrogenic differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: Our data indicate that bone marrow-derived stromal cells of horses can be characterized as MSCs. Equine MSCs have a high transduction rate and migratory potential and adapt to scaffold material in culture. As an autologous cell population, equine MSCs can be regarded as a promising cell population for tissue engineering in lesions of the musculoskeletal system in horses. PMID- 17916019 TI - Comparison of ophthalmic measurements obtained via high-frequency ultrasound imaging in four species of snakes. AB - OBJECTIVE: To measure the dimensions of the eyes of living snakes by use of high frequency ultrasound imaging and correlate those measurements with age, length, and weight. Animals-14 clinically normal snakes. PROCEDURES: Species, age, length, weight, and horizontal spectacle diameter were recorded, and each snake underwent physical and ophthalmic examinations; ultrasonographic examination of both eyes was performed by use of a commercially available ultrasound unit and a 50-MHz transducer. Ultrasonographic measurements included spectacle thickness, subspectacular space depth, corneal thickness, anterior chamber depth, lens thickness, vitreous cavity depth, and globe length. All measurements were made along the visual axis. RESULTS: 2 corn snakes, 5 California king snakes, 1 gopher snake, and 6 ball pythons were examined. There were no significant differences within or between the species with regard to mean spectacle thickness, corneal thickness, or subspectacular space depth. However, mean horizontal spectacle diameter, anterior chamber depth, and axial globe length differed among the 4 species; for each measurement, ball pythons had significantly larger values than California king snakes. CONCLUSIONS AND CLINICAL RELEVANCE: Spectacle thickness, subspectacular space depth, and corneal thickness were similar among the species of snake examined and did not vary significantly with age, length, or weight. Measurements of these dimensions can potentially serve as baseline values to evaluate snakes of these species with a retained spectacle, subspectacular abscess, or subspectacular fluid accumulation. Anterior chamber depth and axial length appeared variable among species, but axial length did not vary with age, length, or weight in the species studied. PMID- 17916020 TI - Pharmacokinetics of voriconazole following intravenous and oral administration and body fluid concentrations of voriconazole following repeated oral administration in horses. AB - OBJECTIVE: To determine the pharmacokinetics of voriconazole following IV and PO administration and assess the distribution of voriconazole into body fluids following repeated PO administration in horses. ANIMALS: 6 clinically normal adult horses. PROCEDURES: All horses received voriconazole (10 mg/kg) IV and PO (2-week interval between treatments). Plasma voriconazole concentrations were determined prior to and at intervals following administration. Subsequently, voriconazole was administered PO (3 mg/kg) twice daily for 10 days to all horses; plasma, synovial fluid, CSF, urine, and preocular tear film concentrations of voriconazole were then assessed. RESULTS: Mean +/- SD volume of distribution at steady state was 1,604.9 +/- 406.4 mL/kg. Systemic bioavailability of voriconazole following PO administration was 95 +/- 19%; the highest plasma concentration of 6.1 +/- 1.4 microg/mL was attained at 0.6 to 2.3 hours. Mean peak plasma concentration was 2.57 microg/mL, and mean trough plasma concentration was 1.32 microg/mL. Mean plasma, CSF, synovial fluid, urine, and preocular tear film concentrations of voriconazole after long-term PO administration were 5.163 +/- 1.594 microg/mL, 2.508 +/- 1.616 microg/mL, 3.073 +/- 2.093 microg/mL, 4.422 +/- 0.8095 microg/mL, and 3.376 +/- 1.297 microg/mL, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that voriconazole distributed quickly and widely in the body; following a single IV dose, initial plasma concentrations were high with a steady and early decrease in plasma concentration. Absorption of voriconazole after PO administration was excellent, compared with absorption after IV administration. Voriconazole appears to be another option for the treatment of fungal infections in horses. PMID- 17916021 TI - Effects of foodborne Fusarium mycotoxins with and without a polymeric glucomannan mycotoxin adsorbent on food intake and nutrient digestibility, body weight, and physical and clinicopathologic variables of mature dogs. AB - OBJECTIVE: To investigate the effects of feeding cereal-based diets that are naturally contaminated with Fusarium mycotoxins to dogs and assess the efficacy of a polymeric glucomannan mycotoxin adsorbent (GMA) in prevention of Fusarium mycotoxicosis. ANIMALS: 12 mature female Beagles. PROCEDURES: Dogs received each of 3 cereal-based diets for 14 days. One diet was uncontaminated (control diet), and the other 2 contained contaminated grains; one of the contaminated diets also contained 0.2% GMA. Contaminants included deoxynivalenol, 15-acetyl deoxynivalenol, zearalenone, and fusaric acid. Food intake and nutrient digestibility, body weight, blood pressure, heart rate, and clinicopathologic variables of the dogs were assessed at intervals during the feeding periods. RESULTS: Food intake and body weight of dogs fed the contaminated diet without GMA were significantly decreased, compared with effects of the control diet. Reductions in blood pressure; heart rate; serum concentrations of total protein, globulin, and fibrinogen; and serum activities of alkaline phosphatase and amylase as well as increases in blood monocyte count and mean corpuscular volume were detected. Consumption of GMA did not ameliorate the effects of the Fusarium mycotoxins. For the GMA-contaminated diet, digestibility of carbohydrate, protein, and lipid was significantly higher than that associated with the control diet, possibly because of physiologic adaptation of the recipient dogs to reduced food intake. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that consumption of grains naturally contaminated with Fusarium mycotoxins can adversely affect dogs' feeding behaviors and metabolism. As a food additive, GMA was not effective in prevention of Fusarium mycotoxicosis in dogs. PMID- 17916023 TI - California court OKs city's declawing ban. PMID- 17916025 TI - Applauds closer ties between human and veterinary medicine. PMID- 17916026 TI - What is your diagnosis? Pectus excavatum. PMID- 17916027 TI - What is your diagnosis? Occipitoatlantoaxial malformation. PMID- 17916028 TI - Risk behaviors for disease transmission among petting zoo attendees. AB - OBJECTIVE: To evaluate risk behaviors for transmission of zoonotic diseases at petting zoos during a period without a recognized disease outbreak. DESIGN: Observational survey with environmental microbiologic sampling. SAMPLE POPULATION: 6 petting zoos in Tennessee. PROCEDURES: Attendees were observed for animal and environmental contact, eating or drinking, hand-to-face contact, and use of a hand sanitizer. Hands were examined via bacteriologic culture on some attendees. Environmental samples were collected at three petting zoos. RESULTS: 991 attendees were observed; of these, 74% had direct contact with animals, 87% had contact with potentially contaminated surfaces in animal contact areas, 49% had hand-to-face contact, and 22% ate or drank in animal contact areas. Thirty eight percent used hand sanitizer; children had better compliance than adults. Results of bacteriologic cultures of hands were negative for Salmonella spp and Escherichia coli O157; Salmonella spp were isolated from 63% and E coli O157 from 6% of the environmental samples. CONCLUSIONS AND CLINICAL RELEVANCE: High risk behaviors were common among petting zoo visitors, and disease prevention guidelines were inconsistently followed. This is an example of the importance of one-medicine, one-health initiatives in protecting the public health. Veterinarians, venue operators, and public health authorities must work together on targeted education to improve implementation of existing disease prevention guidelines. PMID- 17916029 TI - Use of bone morphogenetic proteins for augmentation of bone regeneration. AB - A large body of preclinical and clinical data now documents that recombinant BMPs can be used for skeletal regeneration in humans and animals. Recombinant human BMP-2 and BMP-7 have been approved for use in human patients with long-bone fractures and nonunions and in patients undergoing lumbar fusion or various maxillofacial and dental regenerative procedures. These products have also been made available for veterinary use. PMID- 17916030 TI - Evaluation of cisplatin administered with piroxicam in dogs with transitional cell carcinoma of the urinary bladder. AB - OBJECTIVE: To evaluate the antitumor activity and toxic effects of a conservative dose of cisplatin administered in combination with piroxicam to dogs with transitional cell carcinoma (TCC) of the urinary bladder. DESIGN: Clinical trial (nonrandomized, noncontrolled). ANIMALS: 14 client-owned dogs with histologically confirmed TCC of the urinary bladder. PROCEDURES: Each dog was treated with cisplatin (50 mg/m(2), i.v., q 21 d [reduced to 40 mg/m(2), i.v., q 21 d because of toxic effects]) and piroxicam (0.3 mg/kg [0.14 mg/lb], PO, q 24 h). A CBC, serum biochemical analyses, and urinalysis were performed prior to each cisplatin treatment. Tumor staging (determined from thoracic and abdominal radiographic and urinary bladder ultrasonographic findings) was performed before treatment and at 6-week intervals during treatment. RESULTS: 5 dogs received only 1 dose of cisplatin because of the rapid progression of disease (n = 2) or toxic effects (3). With regard to the neoplastic disease among the other 9 dogs, 1 had partial remission, 5 had stable disease, and 3 had progressive disease after 6 weeks of treatment. Median progression-free interval was 78 days (range, 20 to 112 days). Median survival time was 307 days (range, 29 to 929 days). Moderate to severe renal toxicosis and moderate to severe gastrointestinal toxicosis developed in 5 and 8 dogs, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Because of minimal efficacy and associated renal and gastrointestinal toxicosis, administration of cisplatin (40 to 50 mg/m(2)) with piroxicam cannot be recommended for treatment of dogs with TCC of the urinary bladder. PMID- 17916031 TI - Results of the veterinary enalapril trial to prove reduction in onset of heart failure in dogs chronically treated with enalapril alone for compensated, naturally occurring mitral valve insufficiency. AB - OBJECTIVE: To determine the efficacy of long-term enalapril administration in delaying the onset of congestive heart failure (CHF). DESIGN: Placebo-controlled, double-blind, multicenter, randomized trial. ANIMALS: 124 dogs with compensated mitral valve regurgitation (MR). PROCEDURES: Dogs randomly assigned to receive enalapril or placebo were monitored for the primary endpoint of onset of CHF for < or = 58 months. Secondary endpoints included time from study entry to the combined endpoint of CHF-all-cause death; number of dogs free of CHF at 500, 1,000, and 1,500 days; and mean number of CHF-free days. RESULTS: Kaplan-Meier estimates of the effect of enalapril on the primary endpoint did not reveal a significant treatment benefit. Chronic enalapril administration did have a significant benefit on the combined endpoint of CHF-all-cause death (benefit was 317 days [10.6 months]). Dogs receiving enalapril remained free of CHF for a significantly longer time than those receiving placebo and were significantly more likely to be free of CHF at day 500 and at study end. CONCLUSIONS AND CLINICAL RELEVANCE: Chronic enalapril treatment of dogs with naturally occurring, moderate to severe MR significantly delayed onset of CHF, compared with placebo, on the basis of number of CHF-free days, number of dogs free of CHF at days 500 and study end, and increased time to a combined secondary endpoint of CHF-all cause death. Improvement in the primary endpoint, CHF-free survival, was not significant. Results suggest that enalapril modestly delays the onset of CHF in dogs with moderate to severe MR. PMID- 17916032 TI - Long-term outcome of distal femoral osteotomy for treatment of combined distal femoral varus and medial patellar luxation: 12 cases (1999-2004). AB - OBJECTIVE: To determine long-term outcome of distal femoral osteotomy as a component of treatment for distal femoral varus and medial patellar luxation in large-breed dogs. DESIGN: Retrospective case series. ANIMALS: 12 dogs (16 stifle joints). PROCEDURES: Medical records and radiographs were reviewed to identify large-breed dogs with medial patellar luxation (grade > or = 2) and femoral varus angle > or = 12 degrees treated with distal femoral osteotomy, with a minimum follow-up (by a veterinarian) of 18 months. Signalment, weight, medial patellar luxation and lameness grade, pre- and postoperative femoral varus angle, surgical technique, time to radiographic bone union, and complications were recorded. Follow-up with owners via questionnaire was performed > 18 months after surgery. RESULTS: 16 corrective distal femoral osteotomies were performed with ancillary medial patellar luxation procedures in 12 dogs; 4 dogs had staged bilateral procedures. Mean +/- SD preoperative and postoperative femoral varus angles were 16.3 +/- 4.3 degrees and 3.9 +/- 2.5 degrees , respectively. Mean +/- SD time to radiographic union of the distal femoral osteotomy was 52.6 +/- 13 days. One dog had Kirschner wire migration from the tibial tuberosity. Patellar luxation was not detected after surgery in any dog. Mean +/- SD follow-up by a veterinarian was 1,335 +/- 410 days and by use of an owner questionnaire was 1,497 +/- 464 days. All 10 variables of owner-observed patient comfort and function were significantly improved. CONCLUSIONS AND CLINICAL RELEVANCE: Distal femoral osteotomy in combination with traditional treatment provided predictable osteotomy healing, patellar stabilization, and long-term improvement in patient comfort and function when used to treat combined distal femoral varus and medial patellar luxation in large-breed dogs. PMID- 17916033 TI - Prevalence of and intrinsic risk factors for appendicular osteosarcoma in dogs: 179 cases (1996-2005). AB - OBJECTIVE: To determine the prevalence of appendicular osteosarcoma (OSA) in Greyhounds compared with other breeds and identify potential intrinsic risk factors associated with development of OSA. DESIGN: Retrospective case series. ANIMALS: 179 dogs with primary appendicular OSA. PROCEDURES: Medical records of dogs in which primary appendicular OSA had been diagnosed between 1996 and 2005 were reviewed. Prevalence and crude odds ratios for OSA were calculated for various breeds by comparison with a reference population of mixed-breed dogs. Age and sex were examined as potential risk factors for the 3 breeds with highest prevalence. RESULTS: Breed period prevalence of OSA was highest for Greyhounds (21/339 [6.2%]), followed by Rottweilers (51/969 [5.3%]) and Great Danes (13/297 [4.4%]); all 21 Greyhounds with OSA were identified as having retired from racing. Sex was not identified as a risk factor for OSA in these breeds, but in all 3 breeds, risk of OSA increased with age. Greyhounds were significantly older at the time of OSA diagnosis (mean, 9.9 years) than were Rottweilers (8.3 years) and Great Danes (7.8 years). Rottweilers and Great Danes were more likely to have OSA involving the forelimbs than the hind limbs. The most frequent lesion sites for all 3 breeds were the proximal end of the humerus and distal end of the radius. The proximal end of the femur was also a common site for the Greyhounds. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the present study suggested that Greyhounds, Rottweilers, and Great Danes had an increased risk of developing OSA, compared with mixed-breed dogs. PMID- 17916035 TI - Diagnostic imaging findings and endocrine test results in dogs with pituitary dependent hyperadrenocorticism that did or did not have neurologic abnormalities: 157 cases (1989-2005). AB - OBJECTIVE: To compare imaging findings in dogs with pituitary-dependent hyperadrenocorticism (PDH) that did or did not have neurologic abnormalities. Design-Retrospective case series. ANIMALS: 157 dogs with PDH that did (n = 73) or did not (84) have neurologic abnormalities. PROCEDURES: Medical records were reviewed for the presence and nature of clinical signs of CNS disease, and computed tomographic and magnetic resonance images were reviewed for evidence of a pituitary tumor. RESULTS: 60 of the 84 (71%) dogs without neurologic abnormalities and 48 of the 73 (66%) dogs with neurologic abnormalities had a detectable pituitary tumor. However, 17 of the 84 (20%) dogs without neurologic abnormalities had a pituitary macrotumor (ie, a tumor > or = 10 mm in height), and 41 of the 73 (56%) dogs with neurologic abnormalities did not have a detectable pituitary tumor or had a pituitary microtumor. Vague signs of CNS dysfunction (ie, lethargy, inappetence, and mental dullness) were more specific for detection of pituitary macrotumors than were CNS-specific signs (ie, seizure or blindness). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that there was no apparent relationship between a pituitary tumor and development of neurologic abnormalities in dogs with PDH. In addition, neurologic abnormalities in dogs with pituitary macrotumors were often vague (ie, lethargy, inappetence, and mental dullness). PMID- 17916036 TI - In vitro susceptibility patterns of fungi associated with keratomycosis in horses of the northeastern United States: 68 cases (1987-2006). AB - OBJECTIVE: To determine in vitro susceptibility patterns of fungi associated with keratomycosis in horses in the northeastern United States and compare those patterns with results of studies from other geographic regions. DESIGN: Retrospective case series. ANIMALS: 68 horses with keratomycosis. PROCEDURES: Medical records of horses with a clinical diagnosis of keratomycosis, positive results of corneal fungal cultures, and susceptibility data were reviewed from the years 1987 to 2006. Fungal identification and in vitro antifungal susceptibility test results were recorded. The percentage of susceptible isolates was compared among antifungals for all isolates together and for the most common genera individually. RESULTS: 74 fungal isolates from 68 horses that met inclusion criteria were identified. Aspergillus, Candida, and Fusarium spp were the most frequent isolates. Grouped isolates had the highest percentage of susceptibility to nystatin (87.7%), natamycin (87.5%), and clotrimazole (80.6%). Grouped isolates had the lowest percentage of susceptibility to fluconazole (15.8%) and miconazole (27.5%). Aspergillus spp (> or = 81.0%) were most susceptible to nystatin, clotrimazole, itraconazole, and natamycin. Candida spp (100%) were most susceptible to ketaconazole, natamycin, and nystatin. Fusarium spp (100%) were only consistently susceptible to natamycin. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of in vitro susceptibility testing, nystatin, natamycin, or clotrimazole is recommended for initial topical treatment of keratomycosis in horses from the northeastern United States. Contrary to results of studies of ocular fungal isolates of horses from other regions, Candida spp were identified more frequently and miconazole had lower in vitro efficacy in the present study. PMID- 17916037 TI - Effect of hyperimmunization with an Escherichia coli J5 bacterin in adult lactating dairy cows. AB - OBJECTIVE: To determine the effect of hyperimmunization with an Escherichia coli J5 bacterin on serum IgG2 concentration, incidence of clinical mastitis, and rate of survival to the end of the lactation period (ie, day 305) in adult lactating dairy cattle. DESIGN: Randomized controlled trial. ANIMALS: 1,012 Holstein cows in their second lactation and greater. PROCEDURES: All cows were given 3 doses of the J5 bacterin; cows in the hyperimmunization group were given an additional 3 doses during the first 3 months of lactation. Blood was collected from a small sample of cows to determine anti-J5 IgG2 concentrations. RESULTS: Cows in the hyperimmunization group had higher mean serum anti-J5 IgG2 concentrations than did control cows 28 days after administration of the fourth, fifth, and sixth doses of the J5 bacterin. However, mean serum anti-J5 concentrations during the subsequent lactation were not significantly different between groups. The proportions of cows that developed clinical mastitis were not significantly different between groups. However, control cows were more likely to have severe clinical mastitis than were cows in the hyperimmunization group. The percentage of control cows that remained in the herd to day 305 was significantly lower than the percentage of cows in the hyperimmunization group that did. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that hyperimmunization of mature lactating dairy cattle was associated with increased serum anti-J5 IgG2 concentrations and decreased incidence of severe clinical mastitis, but did not alter survival rate of cows that developed severe clinical mastitis. PMID- 17916038 TI - Fatal gastrointestinal parasitism in goats: 31 cases (2001-2006). AB - OBJECTIVE: To evaluate signalment, history, clinical and pathologic findings, and seasonal weather patterns in association with fatal gastrointestinal parasitism in goats. DESIGN: Retrospective case series. ANIMALS: 152 goats that were > 1 day of age. PROCEDURES: Characteristics including age and counts of coccidia oocysts and trichostrongyle ova (eggs) per gram of feces (EPG) in goats that died because of gastrointestinal parasitism and goats that died because of other causes were compared. Weather data and annual incidence of caprine fatal gastrointestinal parasitism were investigated. RESULTS: Death was attributed to gastrointestinal parasitism in 31 of 152 (20%) goats (median age, 5 months; range, 1 month to 7 years); deaths were attributed to coccidiosis (n = 7 goats; median age, 4 months; median EPG, 2,225), trichostrongylosis (6 goats; median age, 1.25 years; median EPG, 3,700), or dual infection (18 goats; median age, 6.7 months; median EPG, 8,088 coccidia and 5,475 trichostrongyles). Sudden onset of weakness or death was a common historical finding; diarrhea was evident in 15 goats. Common postmortem findings in these goats included cachexia, tissue pallor, poorly formed feces, and mesenteric lymphadenomegaly. Wet weather in spring and summer was associated with increased annual incidence of fatal gastrointestinal parasitism in goats. CONCLUSIONS AND CLINICAL RELEVANCE: Gastrointestinal parasitism is an important cause of death in goats. Clinical signs may not develop until just prior to death, and diagnosis is achieved via parasitologic evaluation of feces and necropsy. Seasonal weather patterns should be considered on an annual basis when designing parasite control programs for goats. PMID- 17916039 TI - Neonatal pig liver-derived progenitors for insulin-producing cells: an in vitro study. AB - Beta (beta)-cell replacement represents an attractive approach for the possible cure of type 1 insulin-dependent diabetes mellitus (IDDM). In a search for potential sources of insulin-secreting cells for IDDM substitution therapy, we have focused on the neonatal pig liver, which is putatively enriched in multipotent stem cells. We then isolated cells measuring 10 to 15 microm in diameter, identified as small cells, characterized by a high proliferation rate and positive staining for immature liver and pancreatic endocrine cell markers (i.e., insulin and pancreatic duodenal homeobox). The ability of these cells to transdifferentiate into pancreatic beta-like cells under culture conditions with exendin-4 (Ex-4) or high glucose concentration was examined. We observed that insulin secretion was not physiological in basal conditions, although it became responsive to glucose after 5 days of exposure to Ex-4. This beta-cell-like phenotype remained physiologically stable, even after stimulus withdrawal. Based on these observations, we contend that the proposed cell and tissue model might offer several advantages as a candidate for substitution cell therapy in IDDM, because the neonatal pig liver seems enriched in cells, with a mixed pancreas liver phenotype, that are easier to purify and grow in culture and are more functional than other beta-like cells upon in vitro single short-term stimulation challenge. PMID- 17916040 TI - In vitro expansion of adipose-derived adult stromal cells in hypoxia enhances early chondrogenesis. AB - Cartilage is an avascular tissue, and chondrocytes in vivo experience a severely hypoxic environment. Using a defined in vitro model of early chondrogenesis, we attempted to enrich for cells with an enhanced ability for chondrogenic differentiation by pre-exposure of mouse adipose-derived adult stromal cells (ADASs) to a hypoxic (2% oxygen) environment. ADASs were subsequently expanded in 2% or 21% oxygen environments, resulting in 2 groups of cells, and then early chondrogenic differentiation was induced at 21% oxygen tension using a 3 dimensional micromass culture system. ADAS chondrogenesis was assessed using Alcian Blue staining for proteoglycans and quantification of sulfated glycosaminoglycans. Osteogenesis of the 2 cell groups was also studied. Two percent oxygen tension profoundly increased the proliferation of ADASs. ADASs expanded in 2% oxygen tension exhibited enhanced early chondrogenic differentiation and diminished osteogenesis, suggesting that the reduced oxygen environment may favor chondroprogenitors. Gene expression analysis suggested that matrix metalloproteinase synthesis was inhibited in cells expanded in 2% oxygen. Furthermore, re-oxygenation of the 2% oxygen-expanded ADASs before differentiation did not significantly affect early chondrogenesis. Thus, priming ADASs with 2% oxygen may have selected for chondrogenic progenitors with an enhanced ability to survive and differentiate. This study is relevant for the future application of cell-based therapies involving cartilage tissue regeneration. PMID- 17916042 TI - The future of cell therapies in the treatment of Parkinson's disease. AB - Parkinson's disease (PD) is a common neurological disorder of the brain which has as a part of its core pathology the progressive degeneration of the dopaminergic nigrostriatal pathway. Therefore, cell therapies that aim to restore this degenerated dopaminergic network represent a promising strategy in helping to cure PD. In this review, the authors start by discussing the progress of research on the use of fetal ventral mesencephalic (VM) tissue in transplantation therapies in PD, both from the clinical and experimental perspectives. Then the issues pertinent to its adoption in the clinic are discussed, including the ethical and practical problems with its use, the varied composition of VM tissue that is implanted with the graft and how this may account for some of the problems seen in the clinical trials using this tissue, especially graft-induced dyskinesia. Finally other promising sources of tissue for PD cell therapy are described, including mesenchymal and embryonic stem cells, before concluding on what is the best approach to the cellular repair of the parkinsonian brain. PMID- 17916041 TI - The potential of muscle-derived stem cells for stress urinary incontinence. AB - The suburethral sling procedures, such as transvaginal tape (TVT), have recently gained popularity for the treatment of stress urinary incontinence (SUI). This TVT procedure can reinforce the weakness of pelvic floor muscles but urethral sphincter deficiency remains. Adult stem cell injection therapy for SUI has recently been at the forefront of the repair of deficient urethral function. Muscle-derived stem cells and adipose-derived stem cells are regarded as candidates for the treatment of SUI because these stem cells can be easily obtained in large quantities under local anesthesia, they have the potential to undergo long-term proliferation, self-renewal and multipotent differentiation, and can serve as a vehicle of releasing neurotrophins, such as nerve growth factor, to repair the deficient urethra. PMID- 17916043 TI - Adult marrow-derived very small embryonic-like stem cells and tissue engineering. AB - A population of CXCR4(+) lin(-) CD45(-) cells that express SSEA, Oct-4 and Nanog has been identified in adult bone marrow. These cells are very small and display several features typical for primary embryonic stem cells such as: i) a large nuclei surrounded by a narrow rim of cytoplasm; ii) open-type chromatin (euchromatin); and iii) high telomerase activity. These cells were named very small embryonic-like stem cells (VSEL-SC). The authors hypothesized that they are direct descendants of the germ lineage. Germ lineage, in order to pass genes on to the next generation, has to create soma and thus becomes a 'mother lineage' for all somatic cell lineages present in the adult body. Germ potential is established after conception in a totipotent zygote and retained subsequently during development in blastomers of morula, cells form the inner cell mass of blastocyst, epiblast and population of primordial germ cells. The authors envision that VSEL-SC are epiblast-derived pluripotent stem cells and could potentially become a less-controversial source of stem cells for regeneration. PMID- 17916044 TI - mRNA stability and cancer: an emerging link? AB - Many oncogenes, growth factor, cytokine and cell-cycle genes are regulated post transcriptionally. The major mechanism is by controlling the rate of mRNA turnover for transcripts bearing destabilizing cis-elements. To date, only a handful of regulatory factors have been identified that appear to control a large pool of target mRNAs, suggesting that a slight perturbation in the control mechanism may generate wide-ranging effects that could contribute to the development of a complex disorder such as cancer. In support of this view, mRNA turnover responds to signalling pathways that are often overactive in cancer, suggesting a post-transcriptional component in addition to the well-recognised transcriptional aspect of oncogenesis. Here the authors review examples of deregulated post-transcriptional control in oncogenesis, discuss post transcriptionally regulated transcripts of oncologic significance, and consider the key role of signalling pathways in linking both processes and as an enticing therapeutic prospect. PMID- 17916045 TI - Cellular uptake and intracellular release are major obstacles to the therapeutic application of siRNA: novel options by phosphorothioate-stimulated delivery. AB - The cellular uptake of oligomeric nucleic acid-based tools and drugs including small-interfering RNA (siRNA) represents a major technical hurdle for the biologic effectiveness and therapeutic success in vivo. Subsequent to cellular delivery it is crucial to direct siRNA to the cellular location where it enters the RNA interference pathway. Here the authors summarise evidence that functionally active siRNA represents a minor fraction in the order of 1% of total siRNA inside a given target cell. Exploiting possibilities of steering intracellular release or trafficking of siRNA bears the potential of substantially increasing the biological activity of siRNA. The recently described phosphorothioate stimulated cellular delivery of siRNA makes use of the caveolar system ending in the Golgi apparatus, which contrasts all other known delivery systems. Therefore, it represents an attractive alternative to study whether promoted intracellular release is related to increased target suppression and, thus, increased phenotypic biologic effectiveness. PMID- 17916046 TI - Tuberculosis subunit vaccines: from basic science to clinical testing. AB - More than 80 years after the introduction of Bacillus Calmette-GuErin, the first tuberculosis vaccine, new vaccines for tuberculosis are finally in clinical trials. The selection of antigens on which new subunit vaccines are based represent the first fulfillment of the promise of proteomics and genomics, and the delivery systems for these antigens are likewise the first fruits of the improved understanding of how the host immune system recognizes pathogens. However, clinical trials are still at Phase I and there remain formidable obstacles to the registration of the first new TB vaccines. Here the authors review the vaccines in clinical trials and discuss the different approaches they take to stimulating immunity to Mycobacterium tuberculosis infection, focusing on recombinant subunit vaccines. The challenges that confront these approaches and how they are being addressed are then discussed. PMID- 17916047 TI - Adjuvants in vaccines and for immunisation: current trends. AB - Vaccines represent one of the most successful strategies in medical science. From the mechanistic perspective, vaccination works by manipulating the immune response through selecting, activating and expanding the memory of B and T cells. To determine the magnitude and quality of immune response, suitable vaccine adjuvants are required; therefore, much effort is going into finding new, effective and non-toxic adjuvant formulations focussed on the activation of key immune targets for inducing a long-term, potent and safe immune response. Significant research is being done in this area, to develop new classes of vaccines for use not only against infectious diseases, but also in the treatment of autoimmune disorders, allergies, chronic inflammatory diseases and cancer. Here the authors review and classify some of the main vaccine adjuvants on the basis of their immunomodulating properties on the immune system. PMID- 17916048 TI - The potential of topical DNA vaccines adjuvanted by cytokines. AB - To improve the efficacy of DNA immunization epidermal Langerhans cells are attractive targets to deliver antigen-encoding plasmid DNA. Topical vaccination with naked plasmid DNA has been shown to induce immune responses, and their potency might be improved by chemical and physical methods aimed to enhance the efficiency of plasmid DNA delivery into the skin. Cytokines have also been evaluated as adjuvants for DNA vaccines because they influence the host immune response. This review focuses on the action of several cytokines tested as molecular adjuvants for DNA vaccines and the combination of them with the DermaVir Patch vaccine. DermaVir vaccine, topically administered under a patch, consists of a plasmid DNA that is chemically formulated into a nanoparticle to support vaccine delivery into epidermal Langerhans cells and to induce antigen specific memory T cells. PMID- 17916049 TI - Natural IgM-mediated innate autoimmunity: a new target for early intervention of ischemia-reperfusion injury. AB - Recent studies showed that innate autoimmunity is an early mechanism for ischemia reperfusion (I/R) injury. Results from different animal models showed that reperfusion of ischemic tissues elicits an acute inflammatory response involving a complement system, which is activated by autoreactive natural IgM. Moreover, ischemia-specific self-targets were identified. In contrast to the unsuccessful attempts in the past to treat I/R injury, targeting natural IgM-mediated innate autoimmunity may open a new avenue for early intervention. PMID- 17916050 TI - Daclizumab. AB - Daclizumab is a humanized monoclonal antibody which binds to the IL-2 receptor on activated lymphocytes and blocks the production of IL-2. Its use is well established in solid organ transplantation as induction therapy, especially in high-risk patients where reduction or delayed dose of standard immunosuppression would be beneficial. It has been used effectively in both 2-dose and 5-dose regimens in conjunction with other standard immunosuppressive agents. The incidence of acute rejection appears reduced without increasing the rates of infection or post-transplant lympho-proliferative disorders. The agent is generally well tolerated in adults and children and there is no need for additional monitoring. Daclizumab has also been used outside the transplant arena in a variety of immune-mediated diseases with limited success. PMID- 17916052 TI - In silico prediction of ADMET properties: how far have we come? AB - There have been considerable advances in the last few years in both the quantity and the quality of in silico ADMET property predictions. Most ADMET properties are now computable, and the accuracy of some of the software predictions for physicochemical properties in particular is close to that of measured data. There is, however, universal agreement that more good experimental ADMET data are needed for use in in silico model development, for models are only as good as the data on which they are based. Many data remain confidential but it is to be hoped that, with projects such as the Vitic toxicity database, being developed by Lhasa Limited, pharmaceutical companies will be prepared to release data to an 'honest broker' on a confidential basis, so that better in silico models can be developed. Incorporation of calculated ADMET properties into drug discovery and development is a multi-factorial problem and really needs a multi-factorial solution. Some progress is being made in this direction and it is hoped that within the foreseeable future software will be available for this purpose. PMID- 17916051 TI - Imatinib mesylate in the treatment of hematologic malignancies. AB - The treatment of hematologic malignancies has been based for many years on chemotherapy and possibly, for the more aggressive forms, stem cell transplantation. In 2001, the signal transduction inhibitor 571 (STI571, imatinib mesylate) was reported to have striking effects in chronic myeloid leukaemia patients. Since then, imatinib became the first molecular-targeted agent approved for the treatment of human cancer and was later on demonstrated to be effective in other malignancies, such as Philadelphia positive acute lymphoid leukemia, hypereosinophilic syndromes, gastrointestinal stromal tumours and more recently, systemic mastocytosis and other myeloprolipherative disease-carrying platelet derived growth factor receptor abnormalities. In this article, the authors review the evidence which led to imatinib approval in the treatment of several of the above mentioned diseases. PMID- 17916053 TI - Maximising use of in vitro ADMET tools to predict in vivo bioavailability and safety. AB - The drastic increase in the costs for discovering and developing a new drug and the high attrition rate of development candidates led to shifting of drug discovery strategy to parallel assessment of comprehensive drug properties along with efficacy. The article reviews the benefits and caveats of implementing comprehensive in vitro tools in early drug discovery and their impact on addressing in vivo ADMET issues. With the proposal of four-barrier profiling paradigm and employment of integrated risk assessment, one can exponentially enhance the predictive power of those in vitro tools by taking into consideration the interplays among those profiling parameters. An 'Exposure Cube' is proposed to promote collective employment of solubility/dissolution, permeability and metabolic clearance to address in vivo exposure and to direct optimization of new chemical entities in drug discovery. PMID- 17916054 TI - Cytochrome P450 reaction-phenotyping: an industrial perspective. AB - It is now widely accepted that the fraction of the dose metabolized by a given drug-metabolizing enzyme is one of the major factors governing the magnitude of a drug interaction and the impact of a polymorphism on (total) drug clearance. Therefore, most pharmaceutical companies determine the enzymes involved in the metabolism of a new chemical entity (NCE) in vitro, in conjunction with human data on absorption, distribution, metabolism and excretion. This so called reaction-phenotyping, or isozyme-mapping, usually involves the use of multiple reagents (e.g., recombinant proteins, liver subcellular fractions, enzyme selective chemical inhibitors and antibodies). For the human CYPs, reagents are readily available and in vitro reaction-phenotyping data are now routinely included in most regulatory documents. Ideally, the various metabolites have been definitively identified, incubation conditions have afforded robust kinetic analyses, and well characterized (high quality) reagents and human tissues have been employed. It is also important that the various in vitro data are consistent (e.g., scaled turnover with recombinant CYP proteins, CYP inhibition and correlation data with human liver microsomes) and enable an integrated in vitro CYP reaction-phenotype. Results of the in vitro CYP reaction-phenotyping are integrated with clinical data (e.g., human radiolabel and drug interaction studies) and a complete package is then submitted for regulatory review. If the NCE receives market approval, information on key routes of clearance and their associated potential for drug-drug interactions are included in the product label. The present review focuses on in vitro CYP reaction-phenotyping and the integration of data. Relatively simple strategies enabling the design and prioritization of follow up clinical studies are also discussed. PMID- 17916055 TI - Use of proteomics for the discovery of early markers of drug toxicity. AB - Toxicity and safety issues remain a significant problem for drug development efforts by pharmaceutical and biotechnology companies. Exisiting early biomarkers of toxicity are insufficient and this is demonstrated by the high failure rate of candidate therapeutics due to toxicity problems. It is anticipated that the advent of 'omic' technologies should facilitate a comprehensive understanding of the perturbation of biological systems by toxic insults and, as such, will lead to better predictive models of toxicity for use in drug development. The field of proteomics continues to develop rapidly and it is already evident that proteomic approaches have much to contribute to the field of 'systems toxicology' and to the development of novel biomarkers of toxicity. Here, the key proteomic approaches are reviewed, their applications in pharmaceutical toxicology are described and what shape future developments in this arena might take is considered. PMID- 17916056 TI - The role of mitochondrial and plasma membrane nucleoside transporters in drug toxicity. AB - Many anticancer and antiviral drugs are nucleoside analogues, which interfere with nucleotide metabolism and DNA replication to produce pharmacological effects. Clinical efficacy and toxicity of nucleoside drugs are closely associated with nucleoside transporters because they mediate the transport of nucleoside drugs across biological membranes. Two families of human nucleoside transporters (equilibrative nucleoside transporters and concentrative nucleoside transporters) have been extensively studied for several decades. They are widely distributed, from the plasma membrane to membranes of organelles such as mitochondria, and the distribution differs in different tissues. In addition, they have different specificities to nucleoside drugs. The characteristics of equilibrative and concentrative nucleoside transporters affect the therapeutic outcomes achieved with anticancer and antiviral nucleoside drugs. In this review, an overview of the role of mitochondrial and plasma membrane nucleoside transporters in nucleoside drug toxicity is provided. Rational design and therapeutic application of nucleoside analogues are also discussed. PMID- 17916057 TI - Drug interactions involving ethanol and alcoholic beverages. AB - Ethanol is likely among the most widely and extensively used drugs in the world. It has also been demonstrated to alter the expression or activity of some drug metabolizing enzymes. Thus, marked ethanol-provoked drug interactions could be of notable clinical importance. To date, relatively few clinically important interactions have been reported, involving cocaine, disulfiram and tacrolimus. Limited or modest interactions with ethanol have also been reported for drugs such as abacavir, cisapride, 'ecstasy' (3,4-methylenedioxymetamfetamine), gamma hydroxybutyrate, methylyphenidate, metronidazole and verapamil. Most of these interactions do not seem to involve CYP2E1, the enzyme initially characterized and cloned based on its ability to metabolize and be induced by ethanol. Important work has elucidated the relationship between CYP2E1-mediated formation of the hepatotoxic metabolite of acetaminophen and alcohol consumption. Lastly, drug interactions involving other components of alcoholic beverages such as flavonoid and other polyphenolic components of red wine have been reported. PMID- 17916058 TI - Erratic absorption of intramuscular antimicrobial delivery in infants and children. AB - In the absence of intravenous or enteral access, an intramuscular (IM) injection of medications is a viable alternative. Vaccines, antibiotics and analgesics are commonly administered by this route in pediatric medicine to enhance adherence to regimens. Although it is expected that the entire dose be administered by the IM route, the absorption (rate and extent) of the medication from the muscle tissue into systemic circulation can be erratic and depends on the drug's physicochemical factors and patient-specific factors. Despite the variability in absorption, administration of medications by the IM route may be considered when medications do not need to attain maximal concentrations rapidly. IM injections are generally well-tolerated, but potential injury to the injected muscle and the proximal nerves should be recognized. PMID- 17916059 TI - The use of escitalopram beyond major depression: pharmacological aspects, efficacy and tolerability in anxiety disorders. AB - Escitalopram, the active (S)-enantiomer of citalopram, has been approved in many countries throughout the world for the treatment of depression and anxiety disorders. It is more potent and selective than citalopram in inhibiting serotonin re-uptake in the CNS, and less potent than various other selective serotonin re-uptake inhibitors in relation to other transporter proteins and receptors: in particular, it is six times less potent than citalopram in binding to the histamine H1 and muscarinic receptors. Escitalopram has favourable pharmacokinetics: it is rapidly absorbed, has a bioavailability of 80% and is not affected by food intake. It has little potential for drug interactions: it has low protein binding and, as it is metabolised by three CYP isozymes, any impairment in the activity of one is unlikely to have a significant effect on metabolic clearance. Caution is necessary only when it is coadministered with drugs metabolised by CYP2D6, such as metoprolol, or administered to the elderly or patients with severe hepatic or renal impairment. The multiple-dose pharmacokinetics of oral escitalopram are proportional at a range of doses including its therapeutic doses. Escitalopram is approved for the treatment of a number of anxiety disorders. It seems to be well tolerated and induces few or no discontinuation symptoms, and may be considered a first-line agent for the pharmacotherapy of obsessive-compulsive disorder, generalised anxiety disorder, panic disorder and social phobia. Further studies are needed to define its activity in impulse control disorders. PMID- 17916060 TI - The role of oseltamivir in the treatment and prevention of influenza in children. AB - The burden of seasonal influenza in children is poorly recognized, in spite of the potential for severe and even life-threatening illness and common secondary complications. Children are a primary reservoir for the spread of influenza to both family members and the community, which imposes a sizeable social and economic strain. Although vaccination is the primary intervention against childhood influenza, the antiviral neuraminidase inhibitors, oseltamivir and zanamivir, provide treatment options. Oseltamivir is administered orally to children aged > 1 year and has been shown to cost-effectively reduce the influenza disease burden and duration of viral shedding. Additionally, oseltamivir postexposure prophylaxis provides protective efficacy for children and families. Oseltamivir has shown excellent tolerability and a low potential for viral resistance in pediatric studies. In the event of an influenza pandemic, oseltamivir is expected to be at the forefront of containment strategies. This article reviews the pharmacology, efficacy and tolerability of oseltamivir as treatment and prophylaxis in children. PMID- 17916061 TI - Nifedipine gastrointestinal therapeutic system: an overview of its antiatherosclerotic effects. AB - Atherosclerosis is the main underlying pathology of cardiovascular disease. Experimental studies in animal models provided early evidence of the antiatherosclerotic effects of nifedipine in reducing and reversing plaque formation and improving endothelial function. Over the past decade, clinical trials, including 'Intervention as a Goal in Hypertension Treatment', 'Prospective Randomized Enalapril Study Evaluating Regression of Ventricular Enlargement', 'Evaluation of Nifedipine and Cerivastatin on Recovery of Coronary Endothelial Function' and 'A Coronary Disease Trial Investigating Outcome with Nifedipine Gastrointestinal System', have further demonstrated that nifedipine gastrointestinal therapeutic system can slow the progression of various markers of atherosclerosis, restore endothelial function, and reduce the incidence of coronary events and the need for coronary interventions. These results are reviewed here, along with the impact they have had on therapy guidelines for patients with hypertension and symptomatic stable angina. PMID- 17916062 TI - Detection and identification of Dehalococcoides species responsible for in situ dechlorination of trichloroethene to ethene enhanced by hydrogen-releasing compounds. AB - Previous studies have shown that Dehalococcoides species are responsible for the anaerobic bioremediation of chloroethene pollution. It has been thought that co operation of several species is required for complete dechlorination to ethene. In the present study, we used quantitative PCR of 16 S rRNA and RDase (reductive dehalogenase) genes to examine species changes and the population of Dehalococcoides species in ground water in which the dechlorination of TCE (trichloroethene) to ethene was enhanced by delivery of hydrogen-releasing compounds. The results have shown that at least two different Dehalococcoides species co-operate in the dechlorination of TCE to ethene. Initially, the number of strains equipped with TCE RDase increased approx. 10(5)-fold. This was followed by a decrease to the original level, according to the exhaustion of TCE. Subsequently, another strain appeared, which had a VC (vinyl chloride) RDase gene similar to bvcA of Dehalococcoides sp. BAV1 and is probably responsible for the dechlorination of VC to ethene. Analysis of several genes has suggested that the former strain is like Dehalococcoides sp. FMC-TCE, and the latter strain is similar to the Dehalococcoides sp. strain that exists in the Dehalococcoides containing mixed culture KB1. These results support the notion that monitoring Dehalococcoides species by the presence of RDase genes as genetic markers provides detailed information on the progress of bioremediation of chloroethenes, which will be useful to improve the efficiency of bioremediation. PMID- 17916063 TI - Activity profiling of human deSUMOylating enzymes (SENPs) with synthetic substrates suggests an unexpected specificity of two newly characterized members of the family. AB - SENPs [Sentrin/SUMO (small ubiquitin-related modifier)-specific proteases] include proteases that activate the precursors of SUMOs, or deconjugate SUMOs attached to target proteins. SENPs are usually assayed on protein substrates, and for the first time we demonstrate that synthetic substrates can be convenient tools in determining activity and specificity of these proteases. We synthesized a group of short synthetic peptide fluorogenic molecules based on the cleavage site within SUMOs. We demonstrate the activity of human SENP1, 2, 5, 6, 7 and 8 on these substrates. A parallel positional scanning approach using a fluorogenic tetrapeptide library established preferences of SENPs in the P3 and P4 positions that allowed us to design optimal peptidyl reporter substrates. We show that the specificity of SENP1, 2, 5 and 8 on the optimal peptidyl substrates matches their natural protein substrates, and that the presence of the SUMO domain enhances catalysis by 2-3 orders of magnitude. We also show that SENP6 and 7 have an unexpected specificity that distinguishes them from other members of the family, implying that, in contrast to previous predictions, their natural substrate(s) may not be SUMO conjugates. PMID- 17916064 TI - Prevalence and predictors of cardiac hypertrophy and dysfunction in patients with Type 2 diabetes. AB - The aim of the present study was to determine the prevalence and predictors of an abnormal echocardiogram in patients with Type 2 diabetes. Cardiac function and structure were rigorously assessed by comprehensive transthoracic echocardiographic techniques in 229 patients with Type 2 diabetes. Cardiovascular risk factors and diabetic complications were assessed, and predictors of an abnormal echocardiogram were identified using multivariate logistic regression analysis. An abnormal echocardiogram was present in 166 patients (72%). LVH (left ventricular hypertrophy) occurred in 116 patients (51%), and cardiac dysfunction was found in 146 patients (64%), of whom 109 had diastolic dysfunction alone and 37 had systolic+/-diastolic dysfunction. Independent predictors of an abnormal echocardiogram were obesity, age, the number of antihypertensive drugs used (all P<0.001) and creatinine clearance (P<0.05). The risk of an abnormal echocardiogram increased by 9% for each year over 50 years of age {OR (odds ratio), 1.09 [95% CI (confidence interval), 1.04-1.15]}, 3-fold if obesity was present [BMI (body mass index) >30; OR, 4.2 (95% CI, 1.9-9.0)] and by 80% for each antihypertensive agent used [OR, 1.8 (95% CI, 1.3-2.4) per agent]. In conclusion, an abnormal cardiac echocardiogram is common in patients with Type 2 diabetes. Importantly, although cardiac abnormalities can be predicted by traditional risk factors, such as age, obesity and renal function, the absence of micro- or macro-vascular complications does not predict a normal echocardiogram. We suggest that an echocardiogram identifies those with Type 2 diabetes at increased cardiovascular risk due to occult LVH and diastolic dysfunction, and this information may lead to more aggressive management of known risk factors in the clinic. PMID- 17916065 TI - High rates of superoxide production in skeletal-muscle mitochondria respiring on both complex I- and complex II-linked substrates. AB - Despite the considerable interest in superoxide as a potential cause of pathology, the mechanisms of its deleterious production by mitochondria remain poorly understood. Previous studies in purified mitochondria have found that the highest rates of superoxide production are observed with succinate-driven reverse electron transfer through complex I, although the physiological importance of this pathway is disputed because it necessitates high concentrations of succinate and is thought not to occur when NAD is in the reduced state. However, very few studies have examined the rates of superoxide production with mitochondria respiring on both NADH-linked (e.g. glutamate) and complex II-linked substrates. In the present study, we find that the rates of superoxide production (measured indirectly as H2O2) with glutamate+succinate (approximately 1100 pmol of H2O2 x min(-1) x mg(-1)) were unexpectedly much higher than with succinate (approximately 400 pmol of H2O2 x min(-1) x mg(-1)) or glutamate (approximately 80 pmol of H2O2 x min(-1) x mg(-1)) alone. Superoxide production with glutamate+succinate remained high even at low substrate concentrations (<1 mM), was decreased by rotenone and was completely eliminated by FCCP (carbonyl cyanide p-trifluoromethoxyphenylhydrazone), indicating that it must in large part originate from reverse-electron transfer through complex I. Similar results were obtained when glutamate was replaced with pyruvate, alpha-ketoglutarate or palmitoyl carnitine. In contrast, superoxide production was consistently lowered by the addition of malate (malate+succinate approximately 30 pmol of H2O2 x min( 1) x mg(-1)). We propose that the inhibitory action of malate on superoxide production can be explained by oxaloacetate inhibition of complex II. In summary, the present results indicate that reverse-electron transfer-mediated superoxide production can occur under physiologically realistic substrate conditions and suggest that oxaloacetate inhibition of complex II may be an adaptive mechanism to minimize this. PMID- 17916066 TI - Validation of spermidine synthase as a drug target in African trypanosomes. AB - The trypanocidal activity of the ODC (ornithine decarboxylase) inhibitor DFMO (difluoromethylornithine) has validated polyamine biosynthesis as a target for chemotherapy. As DFMO is one of only two drugs used to treat patients with late stage African trypanosomiasis, the requirement for additional drug targets is paramount. Here, we report the biochemical properties of TbSpSyn (Trypanosoma brucei spermidine synthase), the enzyme immediately downstream of ODC in this pathway. Recombinant TbSpSyn was purified and shown to catalyse the formation of spermidine from putrescine and dcSAM (decarboxylated S-adenosylmethionine). To determine the functional importance of TbSpSyn in BSF (bloodstream form) parasites, we used a tetracycline-inducible RNAi (RNA interference) system. Down regulation of the corresponding mRNA correlated with a decrease in intracellular spermidine and cessation of growth. This phenotype could be complemented by expressing the SpSyn (spermidine synthase) gene from Leishmania major in cells undergoing RNAi, but could not be rescued by addition of spermidine to the medium due to the lack of a spermidine uptake capacity. These results therefore genetically validate TbSpSyn as a target for drug development and indicate that in the absence of a functional biosynthetic pathway, BSF T. brucei cannot scavenge sufficient spermidine from their environment to meet growth requirements. PMID- 17916067 TI - Side population cells expressing ABCG2 in human adult dental pulp tissue. AB - AIM: To investigate the presence of side population (SP) cells by the Hoechst exclusion method in human adult dental pulp tissue. METHODOLOGY: Human adult dental pulp-derived cells were generated from third molar teeth. The cells were stained with Hoechst 33342 and sorted into SP cells or non-SP cells [main population (MP) cells]. Both cell types were compared with cell growth and RT-PCR analyses. RESULTS: SP cells that express ABCG2, Nestin, Notch-1 and alpha-smooth muscle actin were found at frequencies ranging from 0.67% to 1.02%. This SP profile disappeared in the presence of verapamil. These SP cells expressed dentine sialophosphoprotein and dentine matrix protein-1 when cultured in osteogenic medium. CONCLUSION: Human adult dental pulp tissue contains SP cells that differentiate into odontoblast-like cells. PMID- 17916068 TI - A bio-molecular film ex-vivo model to evaluate the influence of canal dimensions and irrigation variables on the efficacy of irrigation. AB - AIMS: To devise an ex vivo model to test the efficacy of irrigation (static/dynamic) in removing a bio-molecular film from root canal walls. METHODOLOGY: Forty human teeth with single straight canals were randomly allocated to two groups for static (n = 20) or dynamic (n = 20) irrigation. The root canals were prepared to different apical sizes (20, 40) and tapers (0.04, 0.08). The teeth were split longitudinally into two, stained collagen was applied to the canal surfaces and the tooth reassembled in a silicone matrix for dynamic or static irrigation. Digital images of the canal surface were taken before and after irrigation with 9, 18, 27 and 36 mL solution. The percentage of canal surface covered with stained collagen was quantified (ipWin4). The data were analysed using paired t-tests and linear regression models. RESULTS: All the five explanatory variables: 'volume of irrigant used', 'mode of irrigation', 'orientation of open port of needle', 'corono-apical level of canal' and 'root canal dimension' had a significant (P < 0.001) influence on outcome of irrigation. The corono-apical level of canal was the most dominating factor. After irrigation, the apical third had 19.9% and 33.8% less area covered with the bio-molecular film than the middle and coronal thirds respectively. CONCLUSIONS: The stained collagen bio-molecular film could not be removed completely by either static or dynamic irrigation. Factors influencing removal, in rank order of decreasing priority, were: corono-apical level, apical size and taper of canal preparation, and dynamic/static irrigation. PMID- 17916069 TI - The effect of spreader size on fracture resistance of maxillary incisor roots. AB - AIM: To assess the effect of spreader size used during cold lateral compaction of gutta-percha on fracture resistance of roots in maxillary incisor teeth. METHODOLOGY: The crowns of 50 human maxillary incisor teeth having no carious lesions, no fracture or crazing were resected 2 mm coronal to the cemento-enamel junction. The root canals of the teeth were prepared as follows: Group 1: No canal preparation. Group 2: Preparation using the stepback technique to a size 40 master apical file. Group 3: Canal preparation to size 40 and filling with laterally compacted gutta-percha; the first spreader used was equal to size 40. Group 4: Same as group 3 except the first spreader was equal to size 35. Group 5: Same as group 3 except the first spreader was equal to size 25. For each root, a simulated periodontal ligament was prepared. The roots were than mounted in polyester resin and fractured vertically on a universal testing machine (Shimadzu, Tokyo, Japan). The fracture values of teeth were analysed using Kruskal-Wallis and Mann-Whitney U-tests (P = 0.05). RESULTS: The uninstrumented group had the highest fracture resistance; instrumented, but unfilled roots, demonstrated the lowest resistance values (P = 0.009). There were no differences between the uninstrumented group and group 5 in which a size 25 spreader was used during filling. Use of spreaders larger than size 25 caused a significant reduction in fracture resistance of roots (P < 0.05). CONCLUSION: Spreader size used during lateral compaction of gutta-percha can affect the fracture resistance of roots in extracted teeth. PMID- 17916070 TI - Evaluation of the radiopacity of calcium hydroxide- and glass-ionomer-based root canal sealers. AB - AIM: To evaluate the radiopacity of calcium hydroxide-based root canal sealers (Acroseal, Sealapex and Sealer 26), a glass-ionomer-based sealer (Activ GP Sealer) and a zinc oxide and eugenol-based sealer (Intrafill). METHODOLOGY: Five disc-shaped specimens (10 x 1 mm) were fabricated from each material, according to the International Organization for Standardization (ISO) 6876/2001 standard. After setting of the materials, radiographs were taken using occlusal films and a graduated aluminum step-wedge varying from 2 to 16 mm in thickness. The dental X ray unit (GE1000) was set at 50 Kvp, 10 mA, 18 pulses s(-1) and distance of 33.5 cm. The radiographs were digitized and the radiopacity compared with that of the aluminum step-wedge, using WIXWIN-2000 software (Gendex). Data (mm Al) were submitted to ANOVA and Tukey test. RESULTS: Intrafill was the most radiopaque material (7.67 mm Al) followed by Sealer 26 (6.33 mm Al), Sealapex (6.05 mm Al) and Acroseal (4.03 mm Al). Activ GP was the least radiopaque material (1.95 mm Al, P < 0.05). CONCLUSIONS: The sealers evaluated in this study had different radiopacities. However, except for the glass-ionomer-based sealer, all materials had radiopacity values above the minimum recommended by the ISO standard. PMID- 17916071 TI - Bacterial penetration along different root canal filling materials in the presence or absence of smear layer. AB - AIM: To study the effect of the smear layer on the penetration of bacteria along different root canal filling materials and to examine the dentine/sealer and sealer/core material interfaces for the presence of bacteria. METHODOLOGY: A total of 110 human root segments were instrumented to size 80 under irrigation with 1% sodium hypochlorite. Half of the roots were irrigated with a 5-mL rinse of 17% EDTA. Roots with and without smear layer were filled with gutta-percha (GP) and AH Plus sealer (AH), GP and Apexit sealer (AP), or RealSeal cones and sealer (RS). Following storage in humid conditions at 37 degrees C for 7 days, the specimens were mounted into a bacterial leakage test model for 135 days. Survival analyses were performed to calculate the median time of leakage and log rank test was used for pairwise comparisons of groups. The level of significance was set at P = 0.05. Selected specimens were longitudinally sectioned and inspected by scanning electron microscopy for the presence of bacteria at the interfaces. RESULTS: In the presence of the smear layer, RS and AP leaked significantly more slowly than in its absence. In the absence of the smear layer, AH leaked significantly more slowly than RS. SEM results indicated a differential pattern of bacterial penetration among the sealers. CONCLUSIONS: Removal of the smear layer did not impair bacterial penetration along root canal fillings. A comparison of the sealers revealed no difference except that AH performed better than RS in the absence of the smear layer. PMID- 17916072 TI - Tracheal carcinoma ex pleomorphic adenoma: a rare tumour with potential problems in diagnosis. PMID- 17916073 TI - Cyclooxygenase-2 expression correlates with phaeochromocytoma malignancy: evidence for a Bcl-2-dependent mechanism. AB - AIMS: Phaeochromocytomas are rare but potentially life-threatening neuroendocrine tumours of the adrenal medulla or sympathetic nervous system ganglia. There are no histological features which reliably differentiate benign from malignant phaeochromocytomas. The aim of the study was to evaluate cyclooxygenase (COX)-2 and Bcl-2 as tissue-based biomarkers of phaeochromocytoma prognosis. METHODS AND RESULTS: COX-2 and Bcl-2 expression were examined immunohistochemically in tissue from 41 sporadic phaeochromocytoma patients followed up for a minimum of 5 years after diagnosis. There was a statistically significant association between COX-2 histoscore (intensity x proportion) and the development of tumour recurrence or metastases (P = 0.006). A significant relationship was observed between coexpression of COX-2 and Bcl-2 in the primary tumour and the presence of recurrent disease (P = 0.034). A highly significant association was observed between (i) tumour-associated expression of these two oncoproteins (P = 0.001) and (ii) COX-2 histoscore and the presence of Bcl-2 expression (P = 0.002). COX regression analysis demonstrated no significant relationship between (i) the presence or absence of either COX-2 or Bcl-2 and patient survival or (ii) COX-2 histoscore and patient survival. CONCLUSIONS: COX-2 and Bcl-2 may promote phaeochromocytoma malignancy, and these oncoproteins may be valuable surrogate markers of an aggressive tumour phenotype. PMID- 17916074 TI - Role of 14-3-3 proteins in the regulation of neutral trehalase in the yeast Saccharomyces cerevisiae. AB - In higher eukaryotes, 14-3-3 proteins participate in numerous cellular processes, and carry out their function through a variety of different molecular mechanisms, including regulation of protein localization and enzyme activation. Here, it is shown that the two yeast 14-3-3 homologues, Bmh1p and Bmh2p, form a complex with neutral trehalase (Nth1p), an enzyme that is responsible for trehalose degradation and is required in a variety of stress conditions. In a purified in vitro system, either one of the two 14-3-3 yeast isoforms are necessary for complete activation of neutral trehalase (Nth1p) after phosphorylation by PKA. It is further demonstrated that Bmh1p and Bmh2p bind to the amino-terminal region of phosphorylated trehalase, thereby modulating its enzymatic activity. This work represents the first demonstration of enzyme activation mediated by 14-3-3 binding in yeast. PMID- 17916075 TI - Effects of salinity and light on organic carbon and nitrogen uptake in a hypersaline microbial mat. AB - Utilization of dissolved organic matter (DOM) is thought to be the purview of heterotrophic microorganisms, but photoautotrophs can take up dissolved organic nitrogen (DON) and dissolved organic carbon (DOC). This study investigated DOC and DON uptake in a laminated cyanobacterial mat community from hypersaline Salt Pond (San Salvador, Bahamas). The total community uptake of (3)H-labeled substrates was measured in the light and in the dark and under conditions of high and low salinity. Salinity was the primary control of DOM uptake, with increased uptake occurring under low-salinity, 'freshened' conditions. DOC uptake was also enhanced in the light as compared with the dark and in samples incubated with the photosystem II inhibitor 3(3,4-dichlorophenyl)-1, 1-dimethylurea, suggesting a positive association between photosynthetic activity and DOC uptake. Microautoradiography revealed that some DOM uptake was attributed to cyanobacteria. Cyanobacteria DOM uptake was negatively correlated with that of smaller filamentous microorganisms, and DOM uptake by individual coccoid cells was negatively correlated with uptake by colonial coccoids. These patterns of activity suggest that Salt Pond microorganisms are engaged in resource partitioning, and DOM utilization may provide a metabolic boost to both heterotrophs and photoautrophs during periods of lowered salinity. PMID- 17916076 TI - Low amounts of herbivory by root-knot nematodes affect microbial community dynamics and carbon allocation in the rhizosphere. AB - Increased carbon translocation to the rhizosphere via 'leakage' induced by low amounts of plant parasitic nematodes can foster microorganisms. The effects of the root-knot nematode Meloidogyne incognita on microbial biomass (C(mic)) and community structure (phospholipid fatty acids) in the rhizosphere of barley were studied. Inoculation densities of 2000, 4000, and 8000 nematodes were well below the threshold level for plant damage. A (13)CO(2) pulse-labelling was performed to assess the distribution of assimilated (13)C in the rhizosphere. Infection with M. incognita increased the carbon concentration in shoots, and enhanced root biomass slightly. The presence of nematodes did not affect microbial biomass, but significantly changed the allocation of the recent photosynthate. Less plant carbon was sequestered by microorganisms with increasing nematode abundance. Microbial community structure was distinctly altered in the early stages of the plant-nematode interactions. Both, bacteria and fungi, showed a positive response with 2000, and a negative one with 4000 and 8000 M. incognita added. The results suggest that low-level root herbivory still imposes a considerable carbon demand, and that proliferation of microorganisms due to increased rhizodeposition may be short-termed. The carbon flow to rhizosphere microbial communities is likely dependent on the specific nematode-plant association and the developmental stage of the nematode in the host. PMID- 17916077 TI - Saprophytic growth of inoculated Frankia sp. in soil microcosms. AB - The potential of two Frankia strains to grow saprophytically was studied in nonsterile soil microcosms with ground leaf litter of Alnus glutinosa as the sole carbon and nitrogen sources. Strains Ag45/Mut15 and ArI3, which represent two taxonomic subgroups within the Alnus host infection group were inoculated alone, or together to investigate potential competition. Their growth was analyzed by in situ and dot-blot hybridization. A significant increase in cell numbers and filament length was observed during the first 6 weeks after inoculation for strain Ag45/Mut15, both alone and in mixed culture with strain ArI3, followed by a decrease until the end of the study after 12 weeks. The number of filaments remained unchanged. In contrast, the cell numbers and filament length of strain ArI3 were reduced significantly during the first 2 weeks and were undetectable for the remainder of the study. These results were comparable with those obtained in sterile mineral medium amended with leaf litter of A. glutinosa, although reductions in cell numbers and filament length were less pronounced than in soil microcosms. In concomitant control studies without leaf litter amendments for both experimental setups, filaments of both strains could only be detected immediately after inoculation. These results were matched in all experimental setups by concomitant shifts in the rRNA content of both strains, i.e., an immediate decline in the rRNA content for strain ArI3 after inoculation, and an increase in the rRNA content, followed by a late decline during incubation for strain Ag45/Mut15. These results demonstrated that Frankia strain Ag45/Mut15 could grow saprophytically in soil with complex carbon and nitrogen sources such as leaf litter, while the growth of strain ArI3 was not supported. PMID- 17916078 TI - Topiramate: effects on serum lipids and lipoproteins levels in children. AB - The present controlled study aims to evaluate topiramate (TPM) effect on total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein, very low-density lipoprotein, apolipoproteins A1, B and lipoprotein (a). Seventy patients in evolving age suffering from various types of epilepsy, treated with TPM, (age range: 6 months-22 years) were evaluated before and after 12 months of treatment and compared with 110 sex- and age-matched subjects. At baseline, no significant difference was present between controls and children treated with TPM. After a year, the BMI did not show significant change in adults and remained into respective growth curve. No significant difference in lipids and lipoproteins neither between first and second evaluation nor between patients and controls was found. Some intra-group variation has been noticed: whilst controls maintained similar levels, the 70 patients on TPM monotherapy showed a slight decrease in TC, triglycerides and HDL. These fluctuations, however, occurred in the normal range so neither dietary nor pharmacological treatment of hyperlipidaemia after a year of TPM was necessary. PMID- 17916079 TI - Isoform-specific increase of spastin stability by N-terminal missense variants including intragenic modifiers of SPG4 hereditary spastic paraplegia. AB - Hereditary spastic paraplegia (HSP) is a neurodegenerative disorder selectively affecting axons of spinal cord motoneurons. Classical mutations in the most frequent HSP gene SPAST (spastin protein) act through haploinsufficiency by abolishing the activity of a C-terminal ATPase domain or by interfering with expression from the affected allele. N-terminal missense variants have been suggested to represent rare polymorphisms, to cause unusually mild phenotypes, and to aggravate the effect of a classical mutation. We confirm these associations for p.S44L but do not detect two other variants (p.E43Q; p.P45Q) in HSP patients and controls. We show that neither of several disease mechanisms associated with classical SPAST mutations applies to the N-terminal variants. Instead, all three alterations enhance the stability of one of two alternative spastin isoforms. Their phenotypic effect may thus not be mediated by haploinsufficiency but by increasing isoform competition for interacting proteins, substrates or oligomerization partners. PMID- 17916080 TI - Sewage and community shower drains are environmental reservoirs of Fusarium solani species complex group 1, a human and plant pathogen. AB - In two recent studies, clinical isolates in the Fusarium solani species complex (FSSC) were sequenced; one of the most common lineages was FSSC Group 1 (FSSC 1), a phylogenetic species that is synonymous with F. solani f. sp. cucurbitae race 2, a pathogen of cucurbit fruits. FSSC 1 was also identified in sink and shower drains in two hospitals. The environmental sources of FSSC 1 are important for understanding the epidemiology of both human and plant diseases caused by this organism. FSSC 1 was detected in sewage influent at all six tested urban wastewater treatment plants (WWTPs) in California with a concentration ranging from 75 to 413 colony-forming units (cfu) l(-1), a mean of 246 +/- 52 cfu l(-1) and a median of 254 cfu l(-1). During the treatment process, the concentration of FSSC 1 in the solid and liquid fractions diminished. FSSC 1 was detected in five and six of 14 community shower drains by culturing and polymerase chain reaction, respectively, whereas FSSC DNA was detected in all drains. FSSC accounted for 17 +/- 6% (n = 14) of the total fungal DNA in the drains. FSSC 1 was rarely isolated from post-harvest cucurbit fruits and was not found in cucurbit fields in California. PMID- 17916081 TI - Mutational spectrum of delta-globin gene in the Portuguese population. AB - The phenotype of increased Hb A2 typical of beta-thalassaemia (beta-thal) carriers can be reduced to normal or borderline values because of the co inheritance of a delta-globin gene (HBD, MIM #142000) mutation, which may lead to misinterpretation of diagnostic results. To know the spectrum of delta-globin mutations in the Portuguese population we performed a mutational analysis of the delta-globin gene in a group of 51 Portuguese beta-thal carriers presenting microcytosis, hypochromia and a normal/borderline Hb A2 level and in another group of 15 individuals suspected to have delta-globin structural abnormalities. The heterozygosity for the beta(+)IVS-I-6T-->C (HBB:c. 92+6T>C) mutation was the main cause for the mentioned atypical beta-thal carrier phenotype. Furthermore, eight individuals were double heterozygous for one common beta-thal mutation and the delta(+)Cd27G-->T mutation (Hb A2-Yialousa; HBD:c.82G>T). One of them also presented a novel delta-globin gene promoter mutation,-80G-->A (HBD:c.-130G>A), responsible for about 25% decrease of the promoter activity in transient expression assays. One the other hand, in the other group of 15 individuals suspected to have delta-globin structural abnormalities observed by biochemical methods, some known Hb A2 variants were identified - Hb A2' (HBD:c.49G>C), Hb A2 Babinga (HBD:c.410G>A), and Hb A2-Wrens (HBD:c.295G>A), and the novel Hb A2-Fogo [delta64(E8)(Gly-->Ser); (HBD:c.193G>A)]. This novel Hb A2 variant was observed segregating in linkage with Hb E (HBB:c.79G>A) in a three generation family. In conclusion, six different delta-globin mutations were found, being two of them new molecular defects. All delta-alleles identified were found linked to the expected beta-globin cluster haplotype. All mutations caused a low Hb A2 level and through this could lead to misdiagnosis when inherited together with a beta thal allele. PMID- 17916082 TI - Gemtuzumab ozogamicin in combination with attenuated doses of standard induction chemotherapy can successfully induce complete remission without increasing toxicity in patients with acute myeloid leukemia aged 55 or older. AB - BACKGROUND: In this study, the effectiveness and safety of combining gemtuzumab ozogamicin (GO) with an abbreviated schedule of standard induction chemotherapy were assessed in 37 patients (aged > or =55) yr with previously untreated acute myeloid leukemia (AML). METHODS: GO was administered at a dose of 6 mg/m(2) as a single 2-h intravenous infusion on day 1. Following GO, an abbreviated schedule of induction chemotherapy consisting of idarubicin (12 mg/m(2)/d, days 2-4), and N4-behenoyl-1-beta-arabinofuranosyl cytosine (300 mg/m(2)/d, days 2-6) was given. RESULTS: Thirty-seven patients were treated with GO in combination with chemotherapy. Complete remission (CR) and CR with incomplete platelet recovery were achieved in 28 patients (75.7%) and one patient (2.7%) respectively. Two patients (5.4%) died during induction and two patients (5.4%) with grade 4 treatment emergent adverse effects during chemotherapy did not complete induction chemotherapy. The majority of toxicities were mild and manageable. Severe myelosuppression was universal with significantly prolonged thrombocytopenic period. In total, 25 patients who received consolidation treatment, 19 patients remain alive at the time of analysis. Thirteen patients had undergone hematopoietic stem cell transplantation, three are preparing for transplantation and seven are receiving their consolidation chemotherapy course. CONCLUSION: Although only a relatively small number of cases were included in this preliminary study and the follow-up duration was short, frontline GO in combination with attenuated conventional chemotherapy was found to be effective and feasible in elderly patients with AML. PMID- 17916083 TI - Serum transferrin receptor, ferritin, and reticulocyte maturity indices during the first year of life in 'large' preterm infants. AB - BACKGROUND: Preterm infants are at risk of developing iron deficiency; among the iron status and hemopoiesis indices the serum transferrin receptor (sTfr) has been shown to be a useful indicator in assessing iron status, while immature reticulocyte production is regarded as an estimator of erythropoiesis. OBJECTIVE: To investigate age-related changes in iron status infants born 'moderately' preterm, with a gestational age (GA) of 32-36 wk, and identify associations between sTfr and other hematological and biochemical iron indices. DESIGN: Hospital-based prospective, longitudinal study in preterm infants. METHODS: Iron and erythropoiesis parameters were evaluated in 181 formula-fed preterm infants at 2 and 6 wk and 3, 6, 9, and 12 months chronological age. Hemoglobulin (Hb), hematocrit (Hct), mean corpuscular volume (MCV), reticulocytes, serum iron (sFe), serum ferritin (sFer), sTfr, and reticulocyte subpopulations were measured. RESULTS: A total of 756 measurements were performed. After an initial decline, Hb rose from month 3 to 12 of life. SFe and sFer and immature reticulocyte count decreased from the second week to the third month and remained stable thereafter. STfr was lower up to 6 wk and stable from month 3 to 12. Iron deficiency anemia (IDA) was found in 5.5% of infants. In 76 measurements sFer was <12 microg/L, implying storage iron deficiency (SID). A negative correlation was observed between sTfr and other indices of iron status such as Hb, Hct, MCV, sFe, and sFer. Infants with sFer <12 microg/L had lower sTfr than those with sFer >12 microg/L. Reticulocyte production was positively associated with STfr, but this association was dependent on the chronological age of the infant. CONCLUSION: Iron depletion is common in formula-fed preterm (32-36 wk GA) infants between month 3 and 12 of life. STfr appears to be an indicator of iron status in preterm infants during the first year of life. PMID- 17916084 TI - Long-term effects of idiotype vaccination on the specific T-cell response in peripheral blood and bone marrow of multiple myeloma patients. AB - OBJECTIVES: To elucidate long-term effects of idiotype (Id) vaccination on Id specific T cells of multiple myeloma (MM) patients and compare Id-specific T-cell responses of peripheral blood with those of bone marrow (BM). MATERIALS AND METHODS: Id-specific T-cell responses of peripheral blood mononuclear cells (PBMC) were compared with those of BM mononuclear cells (BMMC) in 10 MM patients vaccinated with the Id protein at a median time of 41 months since the last immunization. The PBMC responses at late follow-up were also compared with those during active immunization. The responses were assessed by a proliferation assay, enzyme-linked immunospot (ELISPOT) (gamma-interferon), cytometric bead array (CBA) for secreted cytokines and quantitative real-time polymerase chain reaction (QRT-PCR) for cytokine gene expression. RESULTS: At the late testing time, an Id specific response was detected in PBMC of five patients (ELISPOT, CBA, QRT-PCR) and in BMMC of four patients (CBA, QRT-PCR). A response in both compartments was noted only in three patients. The cytokines gene profile was consistent with a predominance of Th(2) cells [interleukin (IL)-4, IL-5, IL-10]. Comparison of the Id-specific responses of PBMC during active immunization with those at the late follow-up showed that the frequency and magnitude of the responses had decreased significantly by time (proliferation/ELISPOT) (P < 0.02) and shifted at the gene level from a Th(1) to a Th(2) profile (P < 0.05). CONCLUSION: Id-specific T-cells may decline overtime and shift toward a Th(2) response and may be found at a similar frequency of patients in blood and BM. PMID- 17916085 TI - Downregulation of neuropilin-1 in patients with acute myeloid leukemia treated with thalidomide. AB - OBJECTIVE: Neuropilin-1 (NRP-1), a non-tyrosine kinase receptor functioning as a mediator of angiogenesis and neuronal guidance, was recently found to be significantly overexpressed in newly diagnosed acute myeloid leukemia (AML) patients with significant correlation to survival. The role of NRP-1 in refractory or relapsed AML patients and its regulation during anti-angiogenic treatment remain to be elucidated. METHODS: Bone marrow biopsies of 10 patients with refractory or relapsed AML were evaluated for NRP-1 expression by immunohistochemical analysis, and NRP-1 expression level was determined before and after start of thalidomide therapy and correlated to response and growth factor expression. RESULTS: NRP-1 expression was significantly increased in AML patients [median 7 arbitrary units (AU)] when compared with controls (n = 38, median 2.75 AU). Under thalidomide treatment, a marked difference in the course of NRP-1 expression between responders and non-responders was observed, however, without a statistical significance (P = 0.071) being reached. Additionally, a significant correlation of the NRP-1 expression level to microvessel density could be detected under treatment with thalidomide (P = 0.018). CONCLUSION: Our data provide evidence of increased NRP-1 expression in relapsed or refractory AML. Additionally, our results suggest that thalidomide-induced antileukemic properties might at least in part be mediated by NRP-1 downregulation. PMID- 17916087 TI - Mandibular osteosarcoma: a diagnostic pitfall on aspiration cytology of the salivary glands. PMID- 17916088 TI - Tissue verification of stage I sarcoidosis. PMID- 17916086 TI - Krit 1 interactions with microtubules and membranes are regulated by Rap1 and integrin cytoplasmic domain associated protein-1. AB - The small G protein Rap1 regulates diverse cellular processes such as integrin activation, cell adhesion, cell-cell junction formation and cell polarity. It is crucial to identify Rap1 effectors to better understand the signalling pathways controlling these processes. Krev interaction trapped 1 (Krit1), a protein with FERM (band four-point-one/ezrin/radixin/moesin) domain, was identified as a Rap1 partner in a yeast two-hybrid screen, but this interaction was not confirmed in subsequent studies. As the evidence suggests a role for Krit1 in Rap1-dependent pathways, we readdressed this question. In the present study, we demonstrate by biochemical assays that Krit1 interacts with Rap1A, preferentially its GTP-bound form. We show that, like other FERM proteins, Krit1 adopts two conformations: a closed conformation in which its N-terminal NPAY motif interacts with its C terminus and an opened conformation bound to integrin cytoplasmic domain associated protein (ICAP)-1, a negative regulator of focal adhesion assembly. We show that a ternary complex can form in vitro between Krit1, Rap1 and ICAP-1 and that Rap1 binds the Krit1 FERM domain in both closed and opened conformations. Unlike ICAP-1, Rap1 does not open Krit1. Using sedimentation assays, we show that Krit1 binds in vitro to microtubules through its N- and C-termini and that Rap1 and ICAP-1 inhibit Krit1 binding to microtubules. Consistently, YFP-Krit1 localizes on cyan fluorescent protein-labelled microtubules in baby hamster kidney cells and is delocalized from microtubules upon coexpression with activated Rap1V12. Finally, we show that Krit1 binds to phosphatidylinositol 4,5 P(2)-containing liposomes and that Rap1 enhances this binding. Based on these results, we propose a model in which Krit1 would be delivered by microtubules to the plasma membrane where it would be captured by Rap1 and ICAP-1. PMID- 17916089 TI - Cytological criteria of endometrial lesions with emphasis on stromal and epithelial cell clusters: result of 8 years of experience with intrauterine sampling. AB - OBJECTIVE: There are a number of unresolved issues in endometrial cytology. They include the significance of nuclear atypia for the diagnosis of grade1 adenocarcinoma (G1AC) and atypical endometrial hyperplasia (AEH), cytological criteria of endometrial hyperplasia without atypia, and recognition of stromal cell cluster (SC) and its distinction from epithelial cell cluster (EC). METHODS: We examined nuclear atypia, SC and EC in typical cases of five categories: normal endometrium (NEM), simple endometrial hyperplasia without atypia (SEH), complex endometrial hyperplasia without atypia (CEH), G1AC and grade2 adenocarcinoma (G2AC). We classified EC into four types: simple EC (SPEC), large regular EC (LREC), large irregular EC (LIEC) and small irregular EC (SIEC). Based on the results, we developed criteria of endometrial cytology and have evaluated 13 639 cases over 8 years. RESULTS: Nuclear atypia was significantly more frequent in G2AC than in any of the other four categories (P < 0.001). SC was significantly more frequent in NEM and SEH than in the other three categories (P < 0.001). G1AC and G2AC showed significantly higher frequency of LIEC than the other three categories (P < 0.001). CEH exhibited significantly higher frequency of LREC than the four categories (P < 0.001). The sensitivity and the specificity was 88.8% and 99.0% respectively. CONCLUSIONS: We could diagnose G1AC, G2AC and CEH with high accuracy using the established criteria mainly based on SC and EC. We think that the criteria may facilitate an effective screening and an objective interpretation of endometrial samples. PMID- 17916090 TI - Thyroid FNAC cytology: can we do it better? AB - This article reviews recent developments in thyroid fine needle aspiration cytology (FNAC). While thyroid nodules are common, carcinoma is comparatively rare. Although histological assessment is used in most studies as the benchmark, the differential diagnosis on cytology or histology is not always reproducible. The literature shows wide variations in criteria for inadequate thyroid FNAC and study inclusion or exclusion criteria. In-clinic assessment of specimen adequacy and in-clinic reporting of thyroid FNAC has become popular although the costs and resource implications of in-clinic thyroid FNAC assessment and reporting are substantial. Many centres continue to use conventional techniques although liquid based cytology and ultrasound-guided FNAC are gaining in popularity. Standardized categorical systems for FNAC reporting can make results easier to understand for clinicians and give clear indications for therapeutic action. Multidisciplinary case review is also essential, especially when there is diagnostic uncertainty. While currently of limited use, molecular pathology testing holds out some promise for the future. PMID- 17916091 TI - The fine needle aspiration appearances of Kikuchi's lymphadenitis. AB - OBJECTIVE: To describe the fine needle aspiration cytological appearances of Kikuchi's lymphadenitis. METHODS: Cytological review with histological correlation of all cases of Kikuchi's disease (KD) in which there had been an antecedent fine needle aspirate of the involved lymph node prior to lymph node excision between 2001 and 2006. RESULTS: Twelve cases of KD were identified in which cytological and histological material was available. In eight cases the original prospective diagnosis of necrotizing non-granulomatous lymphadenitis consistent with KD had been suggested on the lymph node aspirate. Review of these cytological samples identified abundant extra- and intracellular apoptotic debris - the latter embedded in the cytoplasm of crescentic and phagocytic macrophages, set in a background reactive lymphoid population. Three of 12 cases were initially reported as in keeping with nonspecific reactive lymphadenopathy. Review identified intracellular apoptotic debris but no conspicuous extracellular nuclear debris. One case had originally been reported as possible non-Hodgkin's lymphoma. Histological review of the excised lymph nodes in all 12 cases showed the classical appearances of KD. CONCLUSION: The accurate diagnosis of KD on fine needle aspiration is possible given correct clinical data, an adequately sampled and well-prepared specimen in which the characteristic intra- and extracellular apoptotic nuclear debris with admixed crescentic macrophages are identified on a reactive lymphoid background. PMID- 17916092 TI - Fine needle aspiration cytology of atypical apocrine adenosis of the breast. PMID- 17916093 TI - Fine needle aspiration cytology of the testis as the first-line diagnostic modality in azoospermia: a comparative study of cytology and histology. AB - OBJECTIVES: Male factors are responsible for about half of all infertility cases. Until recently, testicular biopsy was the standard method to ascertain the aetiology of azoospermia. Fine needle aspiration cytology has gained increasing popularity as a simple and minimally invasive procedure that can help in assessing testicular function accurately. This study was aimed at addressing the question whether testicular fine needle aspiration (FNA) may be used as a first line diagnostic modality in azoospermia and to assess its usefulness in the diagnostic protocol. METHODS: The FNA was performed in 78 consecutive azoospermic patients. To obviate sampling errors both testes were aspirated, except when contraindicated. Routine haematoxylin and eosin as well as Romanowsky staining was performed on the smears. RESULTS: The smears were categorized on cytological examination into normal spermatogenesis in 35 (50%) patients, Sertoli cells only syndrome in 22 (31.4%) and maturation arrest at the spermatocyte/spermatid level was seen in 13 (18.4%) patients. There were eight (10.2%) cases with scant smears where cytological diagnosis could not be made. A good correlation between cytological smears and histological sections was found in 54 of 58 testes (93.1%) in which histopathological confirmation was available. CONCLUSIONS: Testicular FNA may be utilized as a first-line investigative modality in patients with azoospermia, provided the procedure is performed and interpreted by experts. PMID- 17916094 TI - Cytological diagnosis of basal cell carcinoma and actinic keratosis, using Papanicolaou and May-Grunwald-Giemsa stained cutaneous tissue smear. AB - OBJECTIVE: Cytology may become the diagnostic method of choice with the advent of new non-invasive treatments for non-melanoma skin cancer, as the sampling technique for cytology entails little tissue disfiguration. The aim of this study was to compare and evaluate the diagnostic performance of scrape cytology using two different cytological staining techniques, and to evaluate additional touch imprint cytology, with that of histopathology of basal cell carcinoma (BCC) and actinic keratosis (AK). METHODS: We investigated 50 BCC and 28 AK histologically verified lesions, from 41 and 25 patients, respectively. Two separate skin scrape samples and one touch imprint sample were taken from each lesion. The smears were stained with Papanicolaou (Pap) or May-Grunwald-Giemsa (MGG) stains. All cytological specimens were examined in random order by pathologists without knowledge of the histology. Cytodiagnostic results were compared with the histopathological report. RESULTS: Scrape cytodiagnosis agreed with histopathology in 48 (Pap) and 47 (MGG) of the 50 BCC cases, and in 26 of 28 (Pap) and 21 of 26 (MGG) AK cases, yielding sensitivities of 96%, 94%, 93% and 81%, respectively. No significant difference in sensitivity between the two staining methods was found but a trend towards higher Pap sensitivity for AK was noted (P = 0.10). Touch imprint cytology confirmed histopathology in 38 of the 77 cases of BCC and AK. CONCLUSION: Cytological diagnosis with either Pap or MGG stain for BCC and AK is reliable, and differentiates well between BCC and AK. Imprint cytology proved to be non-diagnostic in half of the examined cases. PMID- 17916095 TI - The value of calretinin and cytokeratin 5/6 as markers for mesothelioma in cell block preparations of serous effusions. AB - OBJECTIVE: To determine the value of calretinin and cytokeratin (CK) 5/6 in discriminating mesothelioma from adenocarcinoma in serous effusion specimens. METHODS: A total of 101 recent, histologically or clinically confirmed malignant effusions with immunostained cell block preparations were reviewed. The cases consisted of 34 mesotheliomas and 67 adenocarcinomas. This included 17 ascitic fluid and 84 pleural fluid samples. The adenocarcinomas included metastatic carcinomas from the breast (12), lung (19), stomach (3), colon (1), pancreas (2), ovary (6) endometrium (1) and 23 histologically confirmed metastases from unknown primary sites. The cases were assessed as negative or positive (>5% of cells stained). The staining pattern was recorded as cytoplasmic, cell membrane, nuclear or cytoplasmic and nuclear staining. RESULTS: Calretinin staining was present in 97% (33/34) of the mesothelioma cases with a majority of them showing both cytoplasmic and nuclear staining (29/33). Only 3% (2/67) of adenocarcinomas were positive for calretinin, one being a lung adenocarcinoma and the other an adenocarcinoma of unknown primary site in an ascitic fluid. Cytokeratin 5/6 staining was also present in 33/34 (97%) of mesothelioma cases. Six (9%) adenocarcinomas were positive, including metastases from the lung (1), breast (1), ovary (2) and unknown primary site (2). Four of the six adenocarcinoma cases positive for CK5/6 were in ascitic fluids. No cases of mesothelioma were negative for both calretinin and CK5/6. Only one adenocarcinoma case, (which was from unknown primary site in an ascitic fluid sample), was positive for both markers. CONCLUSIONS: The results confirm that calretinin and CK 5/6 are useful markers for mesothelioma in effusion specimens. CK5/6 staining may be less useful for peritoneal fluid specimens where metastatic adenocarcinomas may be more likely to express the antigen. Further study of ascitic/peritoneal specimens is warranted. However, positive staining, particularly for both antigens, is highly indicative of a mesothelial origin for cells. The two markers make a useful addition to EMA and the panel of adenocarcinoma markers routinely applied to effusion specimens. PMID- 17916096 TI - The utility of fine needle aspiration in HIV positive children. AB - OBJECTIVE: To determine the spectrum of disease, diagnostic accuracy and adequacy of fine needle aspirates (FNA) in human immunodeficiency virus (HIV) positive children who present with mass lesions. METHODS: Between January 1997 and December 2002, 95 FNAs were performed in 91 children aged 15 years and younger who were known to be infected with the human immunodeficiency virus (HIV). RESULTS: Head and neck masses including salivary gland swellings were the most common presentation (58.9%) followed by axillary masses (25.3%). Groin masses were aspirated in six children, flank and abdominal masses in four children, buttock masses in three children, a chest wall mass in one child and a sonar guided FNA of a lung mass in one child. Eight FNAs (8.4%) proved inadequate. Reactive lymphadenopathy was diagnosed in 42 cases, mycobacterial infection in 22, four children were diagnosed with abscess, one child had a fungal infection and five were found to have non-Hodgkin's lymphoma. There were four cases each of lymphoepithelial lesion and Kaposi sarcoma. There was one case each of nephroblastoma, rhabdomyosarcoma, myeloma, melanotic progonoma and spindle cells, not otherwise specified. CONCLUSION: Fine needle aspiration in HIV positive children is a worthwhile procedure and in most instances allows a rapid diagnosis obviating the need for surgery and enabling swift treatment to be undertaken where necessary. Ancillary studies form an important diagnostic component. Universal safety precautions must be strictly adhered to. PMID- 17916097 TI - Characterization of de novo microdeletions involving 17q11.2q12 identified through chromosomal comparative genomic hybridization. AB - High-resolution array-comparative genome hybridization (CGH) is a powerful tool for detection of submicroscopic chromosome deletions and duplications. We describe two patients with mild mental retardation (MR) and de novo microdeletions of 17q11.2q12. Although the deletions did not involve the neurofibromatosis type 1 (NF1) gene, they overlap with long-range deletions of the NF1 region which have been encountered in a small group of NF1 patients with more severe MR. Given the overlap of the deletions in our two patients with the large-sized NF1 microdeletions but not with the more frequent and smaller NF1 deletions, we hypothesize that more than one gene in the 17q11.2q12 region may be involved in MR. We discuss candidate genes for MR within this interval that was precisely defined through array-CGH analysis. PMID- 17916098 TI - Differential identification of a rare form of platelet-type (pseudo-) von Willebrand disease (VWD) from Type 2B VWD using a simplified ristocetin-induced platelet-agglutination mixing assay and confirmed by genetic analysis. PMID- 17916099 TI - Progress in understanding and managing natural killer-cell malignancies. AB - The World Health Organization classification of haematolymphoid tumours recognizes three categories of natural killer (NK)-cell neoplasms: blastic NK cell lymphoma, aggressive NK-cell leukaemia, and extranodal NK/T-cell lymphoma, nasal-type. Recent studies indicate that CD4+CD56+ blastic NK-cell lymphoma is of plasmacytoid dendritic cell origin, and true tumours of precursor NK-cell origin may be present mainly in the CD4-CD56+ subset. Myeloid/NK-cell precursor acute leukaemia may also develop from precursor NK cells. However, because the developmental pathway of normal NK cells is not well understood, tumours of precursor NK-cell origin are not clearly identified. Among mature NK-cell tumours, extranodal NK/T-cell lymphoma is relatively common in Asia and Latin America. In localized disease, chemoradiotherapy seems to be promising, and in advanced disease, new combination chemotherapies are under active investigation. Aggressive NK-cell leukaemia is rare and has a poor prognosis. Because NK-cell neoplasms are rare and difficult to manage, rigorous studies are required for their understanding and management. PMID- 17916100 TI - Gene expression profiling of CD34+ cells in patients with the 5q- syndrome. AB - The transcriptome of the CD34+ cells was determined in a group of 10 patients with the 5q- syndrome using a comprehensive array platform, and was compared with the transcriptome of CD34+ cells from 16 healthy control subjects and 14 patients with refractory anaemia and a normal karyotype. The majority of the genes assigned to the commonly deleted region (CDR) of the 5q- syndrome at 5q31-q32 showed a reduction in expression levels in patients with the 5q- syndrome, consistent with the loss of one allele. Candidate genes showing haploinsufficiency in the 5q- syndrome included the tumour suppressor gene SPARC and RPS14, a component of the 40S ribosomal subunit. Two genes mapping to the CDR, RBM22 and CSNK1A1, showed a >50% reduction in gene expression, consistent with the downregulation of the remaining allele. This study identified several significantly deregulated gene pathways in patients with the 5q- syndrome and gene pathway analysis data supports the proposal that SPARC may play a role in the pathogenesis of the 5q- syndrome. This study suggests that several of the genes mapping to the CDR of the 5q- syndrome play a role in the pathogenesis of this disorder. PMID- 17916101 TI - The management of antenatal venous thromboembolism in the UK and Ireland: a prospective multicentre observational survey. AB - This prospective observational study reports on 126 women from 25 UK centres with image-proven antenatal venous thromboembolism (VTE), 62% deep vein thrombosis and 38% pulmonary embolism. Thrombophilia screening was of limited benefit except to identify antithrombin deficiency. Sixteen (13%) patients had previous VTE, all but one was related to previous pregnancy or combined oral contraceptive and 12 received no thromboprophylaxis in the index pregnancy, the other four thus received inadequate low molecular weight heparin (LMWH) doses. Treatment was with dalteparin in 25%, enoxaparin in 47%, tinzaparin in 25% and unfractionated heparin alone in 3%. 66% of patients received once-daily LMWH. Anti-activated factor X (anti-Xa) monitoring was performed at 90% of centres, with a wide range of target values. Thus current management of antenatal VTE, despite widely diverse clinical practice, appeared effective and safe, for there were no recurrent events and postpartum haemorrhage was not increased when compared to known rates. Larger studies are required to confirm this. The need for twice as opposed to once daily LMWH and for anti-Xa monitoring is questioned by this study. The importance of clinical risk assessment and adherence to the Royal College of Obstetricians and Gynaecologists guidelines on antenatal thromboprophylaxis, with adequate LMWH dosing is confirmed. PMID- 17916102 TI - Clinical differences between children and adults with pulmonary hypertension and sickle cell disease. AB - Pulmonary hypertension (PHT) is an important co-morbidity in sickle cell disease (SCD). Despite increasing research in adults, the prevalence and implication of this condition in children is unknown. Charts of 362 SCD patients followed at the Children's Hospital & Research Center Oakland were reviewed to determine clinical variables associated with obtaining echocardiographic screening for PHT, clinical associations of PHT, and associated mortality following diagnosis in adults and children with SCD. In this cohort, patients with underlying lung abnormalities or those on chronic transfusions were more likely to have echocardiograms, however the diagnosis of PHT was often unrecognized. A different clinical phenotype for PHT in adults versus children was identified. Associations with PHT for adults included age, renal and lung disease, hepatitis C, chronic transfusions, and a history of acute chest syndrome (ACS), with ACS being protective. Surprisingly, for children, a history of sepsis, along with a history of ACS, or obstructive lung disease were associated with PHT. Survival analysis found significant mortality for PHT, with a hazard ratio of 17.3 (95% confidence interval 4.9 60.4). The divergent clinical spectrum for PHT between adults and children may point to different age-specific mechanisms or biological expression of PHT. PMID- 17916103 TI - Phase I study of eptifibatide in patients with sickle cell anaemia. AB - The alphaIIbbeta3 antagonist eptifibatide is an effective treatment for patients with acute coronary syndromes (ACS). Platelet reactivity and CD40 ligand (CD40L) may play a role in the pathophysiology of sickle cell anaemia (SCA) similar to that in ACS, suggesting that inhibition of platelet aggregation and CD40L release by eptifibatide may benefit patients with SCA. Following eptifibatide infusion, safety and pharmacodynamic data were obtained from four SCA patients in their non crisis, steady states. Eptifibatide was well tolerated, with no adverse changes in the haematological, biochemical or coagulation parameters studied. Eptifibatide did not increase plasma levels of platelet factor 4 or beta thromboglobulin, P-selectin exposure or platelet:leucocyte aggregate formation. Moreover, decreases in platelet aggregation and soluble CD40L (sCD40L) levels achieved in SCA patients were comparable to those observed in the treatment of ACS. Finally, indicators of inflammation, macrophage inflammatory protein-1alpha, tumour necrosis factor-alpha and myoglobin were reduced following eptifibatide infusion, while vasodilation correlatives, matrix metalloproteinases (MMP-2 and MMP-9) and leptin were increased. Based on these phase I results, eptifibatide may benefit SCA patients by inhibiting platelet aggregation, decreasing sCD40L levels and favourably altering plasma levels of inflammatory mediators. PMID- 17916104 TI - Long-term efficacy and tolerance of rituximab for post-transfusional alloimmune haemolytic anaemia in a thalassaemia patient. PMID- 17916105 TI - Cryofibrinogenaemia. PMID- 17916106 TI - Diagnosis of familial amyloidotic polyneuropathy by bone marrow biopsy. PMID- 17916107 TI - Images in haematology. Hypereosinophilia as a presenting feature of anaplastic large cell lymphoma. PMID- 17916108 TI - Noninvasive markers in the assessment of intestinal inflammation in inflammatory bowel diseases: performance of fecal lactoferrin, calprotectin, and PMN-elastase, CRP, and clinical indices. AB - OBJECTIVES: The aim of this study was to compare the performance of fecal lactoferrin (Lf), calprotectin (Cal), polymorphonuclear neutrophil elastase (PMN e), as well as serum C-reactive protein (CRP) in patients with inflammatory bowel diseases (IBD) to address (a) whether these markers can differentiate IBD patients with endoscopically assessed inflammation from IBD patients without inflammation and from irritable bowel syndrome (IBS); (b) whether they correlate with endoscopic severity of inflammation; and (c) whether a combination of fecal markers with the respective disease-specific activity indices may increase the diagnostic accuracy with reference to the endoscopic severity of inflammation. METHODS: Fecal levels of Lf, Cal, and PMN-e and serum CRP were assessed in 139 patients undergoing diagnostic ileocolonoscopy (54 IBS patients, 42 ulcerative colitis [UC], 43 Crohn's disease [CD]). Disease activity was determined for CU with the colitis activity index (CAI) and for CD with the Crohn's disease activity index (CDAI). The performance of each marker with reference to endoscopic inflammatory activity was assessed by computing correlations, and sensitivity and specificity using published as well as adjusted cutoffs. A comprehensive activity index was computed by combining results from fecal markers, serum CRP, and a clinical activity index. RESULTS: UC or CD patients with active inflammation demonstrated significantly higher levels of Lf, Cal, and PMN-e in feces as well as serum-CRP when compared to patients with inactive inflammation as well as patients with IBS (all P < 0.05). Using adjusted cutoffs enabled a marked improvement of all markers with an overall diagnostic accuracy in IBD of 80.0% for Lf, 80.0% for Cal, 74.1% for PMN-e, 64.0% for CRP, and 79.0% for the respective clinical disease scores. Cal showed the highest diagnostic accuracy in CD (81.4%), whereas Lf was superior to the other markers in UC (83.3%). The comprehensive activity index yielded a further improvement of sensitivity and specificity, with a diagnostic accuracy of 95.3% for UC patients. CONCLUSION: The fecal markers Lf, Cal, and PMN-e are able to differentiate active IBD from inactive IBD as well as from IBS. None of these three stool markers is consistently superior in its ability to reflect endoscopic inflammation, but all three are superior to CRP in their diagnostic accuracy. A combination of the stool markers with the CRP and a disease-specific activity index in a categorical comprehensive activity index can increase the diagnostic accuracy with reference to the endoscopic inflammation in UC. PMID- 17916109 TI - Multicenter, 4-week, double-blind, randomized, placebo-controlled trial of lubiprostone, a locally-acting type-2 chloride channel activator, in patients with chronic constipation. AB - OBJECTIVES: To assess the efficacy and safety of lubiprostone in adults with chronic constipation. METHODS: This multicenter, parallel-group, double-blind controlled trial enrolled 242 patients with constipation and randomized them to receive oral lubiprostone 24 mcg or placebo twice daily for 4 wk. The primary efficacy end point was the number of spontaneous bowel movements (SBMs; those occurring without use of constipation relieving medications) after 1 wk of double blind treatment. Other evaluations included SBMs at weeks 2, 3, and 4; bowel movement (BM) characteristics (i.e., consistency and straining); constipation severity; abdominal bloating/discomfort; global treatment effectiveness ratings; and safety assessments. RESULTS: The 120 lubiprostone-treated patients reported a greater mean number of SBMs at week 1 compared with the 122 placebo-treated patients (5.69 vs 3.46, P= 0.0001), with a greater frequency of SBMs also reported at weeks 2, 3, and 4 (P65; N=477) yielded 256 (53.7%) enrolled subjects, 84 (17.6%) ineligibles, and 120 (25.2%) refusals; 233 of the 256 (87.1%) enrolled patients completed follow-up interviews. INTERVENTION: Nurse training in clinically meaningful use of depression sections of Medicare's mandatory Outcome and Assessment Information Set (OASIS). MEASUREMENTS: Nurse-assessed mood or anhedonia (OASIS) versus research assessments using the Structured Clinical Interview for Axis I Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition Disorders (SCID); referrals for mental health evaluation (agency records), and depression severity (24-item Hamilton Depression Rating Scale; HDRS). RESULTS: Referral rates for patients with (SCID) depressed mood or anhedonia (n=75) varied according to nurse group: 50.0% full intervention, 18.5% minimal, 21.4% control (P=.047). Rates for nondepressed patients (n=180) did not differ (4.9%, 2.0%, 5.8%, respectively; P=.60). In patients with major or minor depression (n=37), referral was associated with symptom improvement. Change in HDRS was 5 points greater in referred patients than others (P=.04). Concordance between OASIS and SCID did not differ between intervention groups. CONCLUSION: TRIAD showed that training nurses to assess for depression using an approach developed in partnership with home healthcare agencies led to appropriate referral and care for depressed patients. PMID- 17916120 TI - Dementia undiagnosed in poor older adults with functional impairment. AB - OBJECTIVES: To identify variables associated with diagnosing dementia in poor older adults by comparing older people with dementia who were diagnosed by their primary care physicians (PCPs) with those not diagnosed by their PCP. DESIGN: Observational study. SETTING: Community-based, in-home cognitive assessment program. PARTICIPANTS: Four hundred eleven adults aged 55 and older with cognitive impairment. MEASUREMENTS: Instrumental activities of daily living (IADLs), activities of daily living (ADLs), Mini-Mental State Examination, Short Blessed Memory Orientation and Concentration Test, and Clinical Dementia Rating. RESULTS: Alzheimer's disease was the most common diagnosis in this group of primarily African-American (73%) older people. Of the 411 participants, 232 (56%) were not diagnosed by their PCP. Participants without a previous diagnosis were older (mean age 81.7 vs 78.7, P=.01), more independent in IADLs (P<.001), and more likely to live alone (P=.001) than persons diagnosed by their PCP. Of the 201 who lived alone, 66% were not diagnosed with dementia by their PCP. Variables associated with PCP diagnosis were more severe cognitive impairment (P<.001), spouse caregiver (P=.009), younger age (P=.02) and care from a university-based PCP (P=.04). CONCLUSION: Persons with dementia who were older and lived alone were less likely to be diagnosed by their PCP. Although persons not diagnosed by their PCP had less cognitive impairment, they had substantial impairment in activities, including handling finances, cooking, and managing medications. PMID- 17916121 TI - Improvement in usual gait speed predicts better survival in older adults. AB - OBJECTIVES: To estimate the relationship between 1-year improvement in measures of health and physical function and 8-year survival. DESIGN: Prospective cohort study. SETTING: Medicare health maintenance organization and Veterans Affairs primary care programs. PARTICIPANTS: Persons aged 65 and older (N=439). MEASUREMENTS: Six measures of health and function assessed at baseline and quarterly over 1 year. Participants were classified as improved at 1 year, transiently improved, or never improved for each measure using a priori definitions of meaningful change: gait speed (usual walking pace over 4 m), 0.1 m/s; Short Physical Performance Battery, 1 point; Medical Outcomes Study 36-item Short Form Health Survey physical function, 10 points; EuroQol, 0.1 point; National Health Interview activity of daily living scale, 2 points; and global health change, two levels or reaching the ceiling. Mortality was ascertained from the National Death Index. Covariates included demographics, comorbidity, cognitive function, and hospitalization. RESULTS: Of the six measures, only improved gait speed was associated with survival. Mortality after 8 years was 31.6%, 41.2%, and 49.3% for those with improved, transiently improved, and never improved gait speed, respectively. The survival benefit for improvement at 1 year persisted after adjustment for covariates (hazard ratio=0.42, 95% confidence interval=0.29-0.61, P<.001) and was consistent across subgroups based on age, sex, ethnicity, initial gait speed, healthcare system, and hospitalization. CONCLUSION: Improvement in usual gait speed predicts a substantial reduction in mortality. Because gait speed is easily measured, clinically interpretable, and potentially modifiable, it may be a useful "vital sign" for older adults. Further research is needed to determine whether interventions to improve gait speed affect survival. PMID- 17916122 TI - The geriatric emergency department. AB - With the aging of the population and the demographic shift of older adults in the healthcare system, the emergency department (ED) will be increasingly challenged with complexities of providing care to geriatric patients. The special care needs of older adults unfortunately may not be aligned with the priorities for how ED physical design and care is rendered. Rapid triage and diagnosis may be impossible in the older patient with multiple comorbidities, polypharmacy, and functional and cognitive impairments who often presents with subtle clinical signs and symptoms of acute illness. The use of Geriatric Emergency Department Interventions, structural and process of care modifications addressing the special care needs of older patients, may help to address these challenges. PMID- 17916124 TI - Depressive symptoms, inflammation, and ischemic stroke in older adults: a prospective analysis in the cardiovascular health study. AB - OBJECTIVES: To investigate the mediator role of inflammation in any relationship between depressive symptoms and ischemic stroke. DESIGN: Longitudinal prospective study. SETTING: Review of medical records, death certificates, and the Medicare healthcare utilization database for hospitalizations. PARTICIPANTS: Total of 5,525 elderly men and women aged 65 and older who were prospectively followed from 1989 to 2000 as participants in the Cardiovascular Health Study. MEASUREMENTS: Depression symptom scores, inflammatory markers. RESULTS: Greater depressive symptoms were associated with risk of ischemic stroke (unadjusted hazard ratio (HR)=1.32, 95% confidence interval (CI)=1.09-1.59; HR=1.26, 95% CI=1.03-1.54, adjusted for traditional risk factors). When a term for inflammation (C-reactive protein (CRP)) was introduced in the model, the HRs were not appreciably altered (unadjusted HR=1.31, 95% CI=1.08-1.58; adjusted HR=1.25, 95% CI=1.02-1.53), indicating that CRP at baseline was not a mediator in this relationship. In analyses stratified according to CRP levels, a J-shaped relationship between depressive symptoms and stroke was evident in the unadjusted analyses; in the fully adjusted model, only CRP in the highest tertile was associated with a higher risk for stroke in the presence of higher depressive symptoms scores. CONCLUSION: The analyses from this prospective study provide evidence of a positive association between depressive symptoms and risk of incident stroke. Inflammation, as measured according to CRP at baseline, did not appear to mediate the relationship between depressive symptoms and stroke. PMID- 17916125 TI - On the identification of clinical Aeromonas by a new restriction fragment length polymorphism of 16S rDNA method. PMID- 17916126 TI - Detection, isolation and characterization of Shiga toxin-producing Escherichia coli in retail-minced beef using PCR-based techniques, immunoassays and colony hybridization. AB - AIMS: To provide information on detection of Shiga toxin-producing Escherichia coli (STEC) in retail-minced beef using an approach combining (i) PCR-based techniques and automated immunoassay for stx screening and detection of the five major serogroups associated with human infection, and (ii) immunomagnetic separation (IMS) and colony hybridization assays for bacterial strain isolation. METHODS AND RESULTS: Twenty-seven out of 164 minced beef samples were stx positive by PCR-ELISA, nine of which were also positive by real-time PCR for at least one marker of the five main serogroups tested (O26, O103, O111, O145 and O157). Two E. coli O103 stx-negative strains were isolated from two out of 10 IMS and nine STEC strains that did not belong to the five main serogroups were isolated by colony hybridization. CONCLUSIONS: PCR techniques are applicable for rapid screening of samples containing both an stx gene and an O-group marker of the five main pathogenic STEC serogroups. Isolation of STEC strains belonging to the main non-O157 serogroups remains difficult. SIGNIFICANCE AND IMPACT OF THE STUDY: This study presents an evaluation of a multi-faceted approach for the detection of the most frequently reported human pathogenic STEC serogroups. The advantages and limits of this strategy are presented. PMID- 17916123 TI - Sleep complaints in community-living older persons: a multifactorial geriatric syndrome. AB - In older persons, sleep complaints in the form of insomnia and daytime drowsiness are highly prevalent and are associated with adverse outcomes. The underlying mechanisms are linked to age-related declines in physiology (normal aging) and age-related increases in disease prevalence (usual aging). This article describes how normal aging leads to less-restorative sleep, characterized by reductions in homeostatic and circadian sleep, and to phase advancement of the sleep-wake cycle, characterized by older persons being more alert in the early morning but drowsier in the early evening. It also describes how usual aging leads to sleep complaints through reductions in health status, loss of physical function, and primary sleep disorders. Psychosocial influences are likewise described, and their relevance to sleep complaints is discussed. These aging-related changes are subsequently incorporated into a conceptual model that describes sleep complaints as a consequence of multiple and interdependent predisposing, precipitating, and perpetuating factors, akin to a geriatric syndrome. The discussion concludes by applying the conceptual model to the sleep-related care of an older person with insomnia and daytime drowsiness and suggesting that the diagnostic assessment consider, in addition to primary sleep disorders, multiple domains, including medical, physical, cognitive, psychological, and social matters, with the intent of developing an overall therapeutic plan and establishing long-term follow-up. PMID- 17916127 TI - Antibiotic resistant Escherichia coli and Salmonella in Russian rooks (Corvus frugilegus) wintering in the Czech Republic. AB - AIMS: To characterize antibiotic resistant Escherichia coli and Salmonella isolates in rooks wintering in the Czech Republic. METHODS AND RESULTS: Three hundred and sixty-three faeces samples from rooks were examined for antibiotic resistant Escherichia coli and Salmonella. Altogether 13.7%E. coli isolates were resistant to antimicrobial agents tested. The dominant type of resistance was to tetracycline. Resistant E. coli isolates were examined for antibiotic resistance genes and class 1 integrons. Five of 29 antibiotic resistant isolates possessed the int1 gene. Nine Salmonella isolates (2.5%) were found in rook faeces. All the isolates belonged to serotype Salmonella enterica serovar Enteritidis phage type PT8 and PT23. CONCLUSIONS: The study suggests that rooks can be infected by antibiotic resistant E. coli and Salmonella isolates, probably reflecting the presence of such isolates in their sources of food and/or water in the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Rooks can serve as reservoirs and vectors of antibiotic resistant E. coli and Salmonella isolates and potentially transmit these isolates over long distances. PMID- 17916128 TI - Variants of eae and stx genes of atypical enteropathogenic Escherichia coli and non-O157 Shiga toxin-producing Escherichia coli from calves. AB - AIMS: To determine the subtypes of stx and eae genes of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic Escherichia coli (EPEC) from calves and to ascertain the typical and atypical nature of EPEC. METHODS AND RESULTS: One hundred and eighty-seven faecal samples from 134 diarrhoeic and 53 healthy calves were investigated for the presence of stx, eae and ehxA virulence genes by polymerase chain reaction and enzyme-linked immunosorbent assay. Subtype analysis of stx(1) exhibited stx(1c) in 13 (31.70%) isolates, while that of stx(2) revealed stx(2c) in eight (24.24%) and stx(2d) in two (6.06%) isolates. Subtyping of eae gene showed the presence of eae-beta, eae-eta and eae-zeta in two, three and four isolates respectively. None of the E. coli isolates possessed stx(2e), stx(2f), eae-alpha, eae-delta, eae-epsilon and eae-xi. All EPEC isolates were atypical. CONCLUSIONS: stx(1), stx(1c), stx(2), stx(2c), stx(2d), eae-beta, eae eta and eae-zeta subtypes are prevalent in STEC and EPEC isolates in India. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first subtype analysis of stx(2) and eae genes of animal E. coli isolates in India and emphasizes the need to investigate their transmission to humans. PMID- 17916129 TI - A novel real-time PCR-based method for the detection of Listeria monocytogenes in food. AB - AIMS: A new real-time PCR-based method was developed for the detection of Listeria monocytogenes in food. METHODS AND RESULTS: A two-step enrichment involving a 24-h incubation in half-Fraser broth followed by a 6-h subculture in Fraser broth was used, followed by cell lysis and real-time PCR with primers and a TaqMan probe previously developed in our laboratory. When the method was evaluated with 144 naturally contaminated food samples, 44 were detected as positive by the PCR-based method and 42 by the standard method EN ISO 11290-1. With 61 food samples artificially contaminated at a level of 10(0) CFU per 25 g, 61 and 58 positive samples were detected by the respective methods. CONCLUSIONS: The developed real-time PCR-based method facilitated the detection of L. monocytogenes in food on the next day after the sample reception, with a reduction of false-positive results because of dead bacterial cells and false negative results because of PCR inhibitors. SIGNIFICANCE AND IMPACT OF THE STUDY: The method can be used for L. monocytogenes detection in food as a faster alternative to current methods. PMID- 17916130 TI - Phenotypic and genotypic diversity of Genista saharae microsymbionts from the infra-arid region of Tunisia. AB - AIMS: Genista saharae, indigenous of Sahara, is a spontaneous shrub that plays an important ecological role for the preservation and fertility of poor and eroded soils. This legume has not been examined for its root nodule bacteria. The taxonomic diversity of bacteria from root nodules of G. saharae growing in the infra-arid region of Tunisia was investigated. METHODS AND RESULTS: A total of 28 bacterial strains isolated from root nodules of G. saharae grown in Tunisian soil were characterized using a polyphasic approach including phenotypic characteristics, PCR-RFLP of 16S rDNA and 16S rRNA gene sequencing. It was found that new isolates are diverse and affiliated to Ensifer (75%), Rhizobium (10%) and Phyllobacterium (15%). The Phyllobacterium isolates lacked the capacity for nodule formation on this plant. CONCLUSIONS: Genista saharae formed nodules with diverse rhizobia in Tunisian soils. Furthermore, our results support the presence of non-nodulating commensal strains (Phyllobacterium) in legumes nodule. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first report on the characterization of G. saharae microsymbionts in Tunisia. PMID- 17916131 TI - Detection of enterotoxigenic Staphylococcus aureus isolates in raw milk cheese. AB - AIM: To develop an easy, rapid and efficient DNA extraction procedure for Staphylococcus aureus detection with a low number of steps and removing completely the PCR inhibitors, applicable to raw milk cheese samples, and to compare phenotypical and genotypical method to detect Staph. aureus isolates and staphylococcal enterotoxins (SEs) production. METHODS AND RESULTS: A total of 33 bovine and caprine raw milk cheese samples were analysed by means of both classic microbiological and molecular techniques. All samples were positive for Staph. aureus contamination. The DNA extraction protocol optimized was found to achieve a detection limit of 100 CFU g(-1) for Staph. aureus. None of the samples tested with immunological assays contained SEs but in 14 of 33 samples a mixture of se positive (sea, sec, sed, seg, sel, sej) isolates were identified. CONCLUSIONS: Staphylococcus aureus is a food-borne pathogen mainly detected in finished dairy products. The rapid and efficient detection of Staph. aureus isolates from dairy products is essential for consumer safety. The direct detection of pathogens from food is possible with careful attention to sample preparation and nucleic acid amplification optimization. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that raw milk cheese samples can be tested for Staph. aureus contamination with a rapid, simple and reproducible procedure. PMID- 17916132 TI - Activity of Nidus Vespae extract and chemical fractions against Streptococcus mutans biofilms. AB - AIMS: To evaluate the effect of Nidus Vespae extract and chemical fractions on the viability and architecture of Streptococcus mutans biofilms. METHODS AND RESULTS: The raw material was first extracted using 95% ethanol/water. Subsequent fractions were prepared from this extract using cyclohexane/ethyl acetate, petroleum ether/ethyl acetate and chloroform/methanol. The biomass dry weight and total protein of samples treated with Nidus Vespae extract and chemical fractions were significantly less than those treated with the vehicle control (P < 0.05). Biofilms treated with Nidus Vespae also resulted in lower percentage of polysaccharide composition. The pH decrease in the biofilm matrix was retarded by Nidus Vespae compared with the vehicle control. Architecture of biofilms treated with Nidus Vespae was different than in the vehicle control and 0.05% chlorhexidine. CONCLUSIONS: Chloroform/methanol fraction was the most effective treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: The significant antibiofilm activity demonstrated by Nidus Vespae shows it to be a promising source of novel anticariogenic agents. PMID- 17916133 TI - Comparison of methods for the detection of methicillin resistance in Staphylococcus aureus isolates from food products. AB - AIMS: To compare several methods for detection of methicillin resistance in Staphylococcus aureus isolates from food. METHODS AND RESULTS: Two hundred S. aureus isolates from food of animal origin were screened for methicillin resistance by a PCR assay specific for the mecA gene, an oxacillin agar screen test and a cefoxitin disk diffusion test. Six out of 200 strains (3%) were found to be methicillin-resistant Staphylococcus aureus (MRSA) by PCR. The oxacillin agar screen test detected only one of the MRSA isolates (sensitivity of 16.7%) and mischaracterized three additional strains as MRSA (specificity of 98.45%). None of the MRSA strains was detected by the cefoxitin test (sensitivity of 0%), while 15 methicillin-susceptible S. aureus (MSSA) strains were misclassified as resistant (specificity of 92.3%). Fifteen MSSA strains displayed a beta-lactamase hyperproducer-like phenotype. The six MRSA (mecA-positive) strains resembled the characteristics of heteroresistant strains. CONCLUSIONS: As MRSA of animal origin may display atypical phenotypes, PCR appears to be more reliable for detection of methicillin resistance in animal strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The study stresses the need for implementing the methods of screening S. aureus from food of animal origin for methicillin resistance. PMID- 17916134 TI - Preventing atrial flutter by preventing inflammation. PMID- 17916135 TI - Anomalous left inferior pulmonary vein crossing midline and draining via common ostium with right inferior pulmonary vein. PMID- 17916136 TI - Ablation of atrial flutter in a patient with situs inversus totalis using integration of real-time three-dimensional electroanatomical mapping. PMID- 17916137 TI - A case of narrow complex tachycardia. PMID- 17916138 TI - Azimilide for the treatment of atrial fibrillation, atrial flutter, and paroxysmal supraventricular tachycardia: results of a randomized trial and insights on the concordance of symptoms and recurrent arrhythmias. AB - INTRODUCTION: Azimilide hydrochloride is an investigational antiarrhythmic medication that had shown evidence of efficacy in prolonging the time to recurrence of atrial fibrillation (AF) or atrial flutter (AFL) and paroxysmal supraventricular tachycardia (PSVT). This study was designed to confirm efficacy of 125 mg daily azimilide. METHODS AND RESULTS: The primary endpoint was ECG documented recurrence of AF, AFL, or PSVT, followed for a maximum of 180 days. Four hundred eighty-two patients were enrolled in the United States and Canada (422 with AF or AFL and 60 with PSVT). The primary efficacy analysis included 402 patients with AF-AFL and 56 patients with PSVT. There was no significant difference in the time to first recurrence of symptomatic supraventricular arrhythmia in the AF-AFL stratum (median of 38 days for azimilide versus 27 days for placebo; hazard ratio [HR] of 1.14, P = 0.29). Similarly, there was no difference in time to recurrence in the PSVT stratum (>180 days for azimilide versus 135 days for placebo; HR = 1.28, P = 0.55). There were three deaths in the azimilide group and one in the placebo group. Four patients had nonsustained ventricular tachycardia (one torsades de pointes), all in the azimilide group. Asymptomatic recurrence was frequent in the AF-AFL group (8% with azimilide and 11% with placebo), but was absent in the PSVT group. False recurrence was common in both groups. CONCLUSION: Azimilide 125 mg daily was not associated with significant prolongation of the time to recurrent symptomatic supraventricular arrhythmias. There was substantial discordance between symptoms and recurrence. PMID- 17916139 TI - High diagnostic yield and accuracy of history, physical examination, and ECG in patients with transient loss of consciousness in FAST: the Fainting Assessment study. AB - BACKGROUND: Transient loss of consciousness (TLOC) is a common clinical problem. OBJECTIVE: The aim of this study was to assess the yield and accuracy of the initial evaluation, consisting of standardized history, physical examination, and ECG performed by attending physicians in patients with TLOC. METHODS AND RESULTS: Five hundred and three adult patients (mean age 53 +/- 19; 56% male) presenting with TLOC to the Academic Medical Center Amsterdam between February 2000 and May 2002 were included in this study. After initial evaluation, the physician made a certain, a highly likely (>80% certain), or no initial diagnosis. Initially undiagnosed patients received additional cardiological testing, additional history taking, and autonomic function tests. After 2 years of follow-up, an expert committee determined the final diagnoses. Two-year follow-up was obtained in 99% of the patients. The yield of certain diagnoses after the initial evaluation was 24%, increasing to 63% after including the highly likely diagnoses. The diagnostic accuracy of the initial certain diagnoses was 93% (95% CI 87-97%), decreasing to 88% (95% CI 84-91%) after inclusion of the initial highly likely diagnoses. CONCLUSION: Attending physicians can make a diagnosis based on initial evaluation in 63% of patients with TLOC, with an overall diagnostic accuracy of 88%. The use of additional testing, beyond history, physical examination, and ECG can be avoided in many patients with TLOC. PMID- 17916140 TI - Electrocardiographic optimization of interventricular delay in cardiac resynchronization therapy: a simple method to optimize the device. AB - INTRODUCTION: Echocardiography is widely used to optimize CRT programming, but it is time-consuming. This study aimed to correlate the optimal interventricular pacing (V-V) interval obtained by echo with the optimal V-V interval obtained by a simpler method based on the surface ECG. METHODS AND RESULTS: Three V-V intervals were tested: LV preactivation at -30 ms, simultaneous biventricular pacing (0 ms), and RV preactivation at +30 ms. The one that achieved the best LV synchrony was chosen as the optimal V-V. This result was then compared with two different ECG measurements. The first ECG method considered the best V-V to be that which achieved the narrowest QRS. The second V-V method consisted in measuring the interval from the pacing spike to the beginning of the fast deflexion of the QRS complex in leads V1, V2, first pacing from the LV (T1), and after from the RV (T2). The T2-T1 interval was considered as a surrogate measurement of interventricular delay and defined as the best V-V. A cohort of 31 consecutive patients treated with CRT was studied. Optimal V-V interval obtained by echo was -30 ms in 25 patients (80%), +30 ms in three patients (10%), and 0 ms in the remaining three patients (10%). Echo results had 32% coincidence with the first ECG method (r = 0.2, P = NS) and 83% coincidence with the second ECG method (r = 0.81 P< 0.001). CONCLUSIONS: The time difference in the fast ventricular depolarization observed between RV and LV stimulation in the surface ECG shows a good correlation with the V-V optimization chosen according to echo. PMID- 17916141 TI - Do the left atrial substrate properties correlate with the left atrial mechanical function? A novel insight from the electromechanical study in patients with atrial fibrillation. AB - BACKGROUND: The atrial substrate is the determinant of occurrence and maintenance of atrial fibrillation (AF), which can induce remodeling of atrial function and structure. This study investigated the relationship between the left atrial (LA) substrate properties and LA mechanical function. METHODS: Forty-four consecutive patients (50.3 +/- 10.7 years old, 33 men) who presented with sinus rhythm during echocardiographic study before receiving catheter ablation for AF were enrolled. The LA diameter, LA volume, ratio of early and late transmitral filling flow velocities (E/A), LA appendage flow velocity, and transmitral velocity-time integral (VTI) were measured by the echocardiography. The LA empty fraction (LAEF), which was obtained via dividing the difference between maximal and minimal LA volume by maximal LA volume, was calculated as a parameter of the global LA contractile function. The LA global contact voltage mapping (NavX system) was performed before pulmonary vein isolation. RESULTS: Mean LA voltage and LA low voltage zone index (LVZ index, area with voltage < 0.5 mV, divided by total LA surface area) showed significant correlation with LA diameter and volume, but only the LA LVZ index showed significant correlation with A-wave velocity, transmitral A-wave VTI, and LAEF (r =-0.340, -0.411, -0.426; P = 0.024, 0.006, 0.005, respectively). We divided the LA LVZ index into three groups (< 10%, 10-20%, > 20%). The LAEF got worse and the transmitral A-wave VTI percentage (divided by transmitral VTI) decreased as LA LVZ index increased. CONCLUSIONS: The LA substrate properties showed close correlation with LA size, but only the LA LVZ index correlated with the LA mechanical function. PMID- 17916142 TI - Electrophysiologic and anatomic characterization of sites resistant to electrical isolation during circumferential pulmonary vein ablation for atrial fibrillation: a prospective study. AB - BACKGROUND: Catheter ablation (CA) by wide encirclement of pulmonary veins (WEPV) restores sinus rhythm in up to 95%. Complex PV-left atrial (LA) connections make achieving electrical isolation (EI) challenging. We examined anatomical and technical features associated with resistance to EI during WEPV in a prospective study. METHODS: One hundred one consecutive patients with symptomatic AF underwent first-time CA guided by electroanatomic mapping and CT integration (Cartomerg). Following double-transseptal access, WEPV was performed. After completion of PV encirclement, the line was mapped and where no signal could be obtained, CA was performed inside the WE line at the site of earliest PV breakthrough on the circular mapping catheter. Sites of EI were tagged. Anatomic studies of corresponding regions of the venoatrial junction in 24 adult hearts were performed. RESULTS: Sites resistant to EI were located at the inferior quadrant (P < 0.001) for the RSPV, superior quadrant (P < 0.001) for the RIPV, and the inferior and anterior quadrants (P < 0.001) for the LSPV. EI was significantly less frequent at the posterior quadrant (P < 0.001) for the LIPV. To achieve EI, CA was necessary inside the WE on the intervenous ridge on the right in 51% and on the left in 41%. The LPV/LAA ridge was investigated by anatomic studies that demonstrated considerable variation in the narrowest width (3-23.7 mm) and transmural thickness (1-5 mm). CONCLUSION: Sites of EI after WEPV have a preferential distribution determined by anatomic features. CA on the intervenous ridge is required in a significant proportion of patients to achieve EI. Atrial folds and ridges increase myocardial thickness creating technical and anatomic challenges for achieving transmural lesions. PMID- 17916143 TI - Autonomic mechanism to explain complex fractionated atrial electrograms (CFAE). AB - OBJECTIVE: To simulate complex fractionated atrial electrograms (CFAE) during sustained atrial fibrillation (AF) in experimental animals. BACKGROUND: The mechanism(s) underlying CFAE has not been fully elucidated. METHODS: Twenty-two dogs were subjected to a right and/or left thoracotomy. A gauze patch soaked with acetylcholine (ACh) was placed on the right atrial appendage (RAA) to induce sustained AF. During AF, varying concentrations of ACh (1, 10, 100 mM) were "painted" on the RA where electrograms showed regular organized activity. In another six dogs, anterior right ganglionated plexi (ARGP) near the sino-atrial node and inferior right GP (IRGP) at the junction of inferior vena cava and atria were sequentially ablated. In five dogs, ACh was injected into ARGP to induce CFAE. RESULTS: During sustained AF, local "painting" with ACh 1 mM and 10 mM induced intermittent CFAE in 1 of 11 and 10 of 11 dogs, respectively. With 100 mM ACh, all 11 showed CFAE (two intermittent, nine continuous). In six other dogs, continuous CFAE induced by topical application of 100 mM ACh were markedly attenuated by ARGP + IRGP ablation. In another five of five dogs, ACh injection into ARGP induced a gradient of CFAE with the continuous CFAE always occurring near the ARGP and CFAE also occurring at left pulmonary vein-atrial junctions. During ARGP ablation, AF was terminated in all five dogs immediately after regularization of the rotor-like electrograms or continuous CFAE. CONCLUSIONS: This study demonstrates an autonomic basis for CFAE formation, suggesting that graded hyperactive states of the autonomic nervous system (ANS) may induce various types of CFAE observed clinically. PMID- 17916144 TI - Coming full circle: contouring the right ventricle in arrhythmogenic right ventricular dysplasia/cardiomyopathy. PMID- 17916145 TI - Learning while burning revisited. PMID- 17916146 TI - Broad complex tachycardia: a broad differential. PMID- 17916147 TI - Early recurrence of ventricular fibrillation after successful defibrillation during prolonged global ischemia in isolated rabbit hearts. AB - INTRODUCTION: The mechanisms that lower the efficacy of electrical defibrillation during prolonged global ischemia remain unclear. METHODS AND RESULTS: Epicardial activation patterns during attempted electrical defibrillation were studied in 18 Langendorff-perfused rabbit hearts at baseline, after 5-minute no-flow global ischemia and after 10-minute reperfusion. DFT(50) (voltage required to achieve 50% probability of successful defibrillation) was determined at each stage. Defibrillation was considered successful if postshock sinus/idioventricular rhythm was present. Prolonged global ischemia converted type 1 VF (multiple wandering wavelets) into type 2 VF (repetitive epicardial breakthroughs, REBs). The mean DFT(50) after 5-minute ischemia (96 +/- 39 V) was significantly lower when compared with that at baseline (154 +/- 47 V, P < 0.0001) and after 10 minute reperfusion (145 +/- 47 V, P < 0.001). However, the incidence of early (within 10 seconds) VF recurrence after successful shock during prolonged global ischemia (23 of 78, 29.5%) was much higher than that at baseline (2 of 60, 3.3%) and after 10-minute reperfusion (5 of 63, 7.9%; P < 0.0001). Mapping data showed that the VF wavefronts during prolonged global ischemia were initially halted by the shock, followed by one to five ventricular escape beats. These beats then triggered REBs and early VF recurrence. In eight out of 11 episodes, the REBs before and after successful shock arose from the same location near the interventricular septum. CONCLUSIONS: There is a significant reduction of DFT(50) during prolonged global ischemia. However, defibrillation appears to fail when the preexisting REBs near the interventricular septum induce early VF recurrence. Shock per se cannot eliminate the substrates of these REBs. PMID- 17916148 TI - Computer-simulated alternative modes of U-wave genesis. AB - OBJECTIVE: Several hypotheses for the origin of the U wave in electrocardiograms have been proposed. We have set out to explore and test alternative modes for U wave genesis via computer simulations. METHODS AND RESULTS: A spatial model of a left ventricle has been constructed from 12 layers composed of cubic cells. Each cell is assigned its own time-dependent action potential with its own contribution to the electrical potential at arbitrary points where ECGs are measured. Simulated ECGs show that U waves can be generated using various combinations of action potentials (APs) across the different layers of the ventricular wall. We demonstrate a new mode of U-wave genesis, even with small differences in the repolarization. CONCLUSION: The U wave can be generated in the presence of strong intercellular coupling. Myocardial layers with prolonged action potentials, like M cells, are not necessarily needed for U-wave genesis. PMID- 17916149 TI - A better mousetrap? The search for an improved CRT optimization technique. PMID- 17916150 TI - DP+1: another simple endpoint for atrial flutter ablation. AB - BACKGROUND: When double potentials (DP) on the line of block are difficult to see, we propose another simple method to verify complete bidirectional block (CBDB) at the end of an atrial flutter ablation. We measured the interval between the electrograms immediately on either side of the line of block on a multipole catheter spanning the isthmus. We called this interval "DP+1" because it is one pair of electrodes away from the DP on the line of block. METHODS: Fifty consecutive patients (age 62 +/- 13 years, LVEF 54 +/- 11%, mean cycle length 241 +/- 34 ms) underwent an atrial flutter ablation using a duodecapolar catheter with 2-10 mm spacing with the distal tip inserted into the mid-coronary sinus and the rest of the poles spanning the isthmus and the low lateral right atrium. Radiofrequency ablation was performed using a 10-mm tip electrode (EP Technologies). The ablation endpoint was the creation of a craniocaudal activation pattern of the opposite wall to the pacing site (septal and lateral of the line of block). RESULTS: The ablation endpoint was achieved in 48 of 50 (96%) patients with 8 +/- 2 RF applications. Adequate DP were found in only 22 of 50 patients (44%), but the DP+1 interval was measurable in all patients. When no block was present, the DP+1 interval was 81 +/- 10 ms, and 160 +/- 18 ms when complete bidirectional block was present (P < 0.001). A DP+1 interval of >140 ms had 100% specificity, 96% sensitivity, 100% positive predictive value for verifying complete bidirectional block. After a follow-up of 528 +/- 253 days, there were no recurrences of AFL, but there were four recurrences of AF (8%). CONCLUSION: When DP cannot be seen, another simple method for verifying CBDB in ablation of typical atrial flutter is a DP+1 interval > 140 ms. PMID- 17916151 TI - Clinical significance of macroscopic T-wave alternans after sodium channel blocker administration in patients with Brugada syndrome. AB - INTRODUCTION: Macroscopic T-wave alternans (TWA) is sometimes observed after sodium channel blocker administration in patients with Brugada syndrome (BS), but little is known about the association between occurrence of TWA and clinical characteristics in BS patients. We investigated the association between spontaneous ventricular fibrillation (VF) occurrence and TWA after pilsicainide, a sodium channel blocker administration in BS patients. METHODS AND RESULTS: We administered pilsicainide at a dose of 1 mg/kg to 77 BS patients (76 males and one female; mean age, 48.4 years) and examined the association between TWA after pilsicainide administration and clinical characteristics, including age, spontaneous VF, syncope, family history of sudden death, spontaneous coved ST elevation, late potentials (LP), induction of VF by programmed electrical stimulation, and SCN5A mutation. None of the patients had TWA before pilsicainide administration, but TWA became apparent in 17 (22.1%) of the patients after pilsicainide administration. Patients with TWA had a significantly higher incidence of spontaneous VF (52.9% vs 8.3%, P < 0.001) and syncope (58.8% vs 26.7%, P < 0.05) than did patients without TWA. Then, we focused on the association between spontaneous VF and clinical characteristics. Patients with spontaneous VF had a significantly higher incidence of TWA (64.3% vs 12.7%, P < 0.001) and LP positive (92.9% vs 56.5%, P < 0.01) than did patients without spontaneous VF. In multivariate analysis, TWA (P = 0.001) and LP (P = 0.047) appeared as the independent predictor for spontaneous VF. CONCLUSION: TWA after pilsicainide administration is associated with a high risk of clinical VF in patients with BS. PMID- 17916152 TI - Genetic basis of hypertrophic cardiomyopathy: from bench to the clinics. AB - Hypertrophic cardiomyopathy (HCM) is a common inherited cardiac disorder that characterized by marked thickening of the left ventricular wall that occurs in the absence of increased external load. HCM is the most common cause of sudden cardiac death under 35 years and in addition causes heart failure. HCM is usually inherited as an autosomal dominant mutation in genes that encode protein constituents of the sarcomere. To date, more than 450 different mutations have been identified within 13 myofilament-related genes. This review focuses current research involved in the discovery of other causative genes, investigation of the mechanisms by which sarcomere genes mutations produce hypertrophy and arrhythmia, and identification of modifying factors that influence clinical expression in HCM patients. The clinical implications of molecular advances in HCM are discussed. PMID- 17916153 TI - Anatomic location of pulmonary vein electrical disconnection with balloon-based catheter ablation. AB - INTRODUCTION: Balloon-based catheters are an emerging technology in catheter ablation for atrial fibrillation, which aim to achieve consistent and rapid ablation encirclement of pulmonary veins (PVs). Recent emphasis has been placed on achieving more proximal electrical isolation within the PV-left atrial (LA) junction. We sought to evaluate the precise anatomic level of PV electrical disconnection with current design balloon-based catheters. METHODS AND RESULTS: Thirteen patients with drug-refractory paroxysmal atrial fibrillation undergoing balloon catheter ablation with the endoscopic laser system (CardioFocus) or the high frequency-focused ultrasound system (ProRhythm) underwent electroanatomic mapping (EAM) of the left atrium. Intracardiac echocardiographic (ICE) imaging was used for visualization of the position of the balloon catheter during energy delivery. Detailed point analysis of the location of electrical disconnection was then documented on EAM and with ICE. Successful electrical isolation was achieved in all 52 PVs. Despite ICE imaging confirming balloon catheter position at the antrum of the PVs, the location of electrical disconnection was demonstrated to be at or near the tubular ostium of the PVs on EAM and on ICE in all patients. CONCLUSION: Current generation balloon-based catheter ablation achieves electrical isolation distal in the LA-PV junction. This may limit the results of such systems in treating nonparoxysmal forms of atrial fibrillation. PMID- 17916154 TI - The transmural activation sequence in porcine and canine left ventricle is markedly different during long-duration ventricular fibrillation. AB - BACKGROUND: Humans are more similar in transmural Purkinje and cardiac ion channel distributions to dogs than pigs. The Purkinje network in pigs is transmural but confined to the endocardium in dogs. Little is known about intramural activation during long-duration ventricular fibrillation (LDVF) given these differences. We tested the hypothesis that the transmural activation sequence is similar in sinus rhythm (SR) and LDVF in dogs as well as pigs, but different between species. METHODS AND RESULTS: In six pigs and seven dogs, 50-60 plunge needles (six electrodes, 2-mm spacing) were placed throughout the left ventricle. Unipolar recordings were made for >10 minutes of LDVF. SR and LDVF activation times were grouped into waves by linking activations along each needle. Origin (earliest activation) and propagation direction were determined for each wave. The mean wave origin was significantly more endocardial in dogs than pigs for SR and 1 through 10 minutes of LDVF. Predominant propagation direction in LDVF and SR was endocardial to epicardial in dogs, but the opposite or equal in both directions in pigs. Fastest activation rate was epicardial in pigs, but endocardial in dogs with an increasing endocardial-to-epicardial activation rate gradient as LDVF progressed in dogs but not pigs. CONCLUSIONS: The transmural activation sequence in SR and LDVF is markedly different between pigs and dogs. These differences may be related to differences in Purkinje fiber and ion channel distributions and suggest that dogs are a better model for investigating activation sequences during LDVF, given the similarities with humans. PMID- 17916155 TI - Effect of atrial electrical remodeling on the efficacy of antiarrhythmic drugs: comparison of amiodarone with I(Kr)- and I(to)/IKur-blockade in vivo. AB - INTRODUCTION: Amiodarone is the gold standard in the prevention of recurrence of atrial fibrillation (AF), but the causes for its superior clinical efficacy are not understood. We hypothesized that atrial electrical remodeling increases the atrial efficacy of amiodarone. METHODS AND RESULTS: We investigated the effect of an acute intravenous dose of amiodarone on atrial refractory periods (AERP) in sinus rhythm (SR) and after 5, 24, and 72 hours of atrial tachypacing in comparison with the I(Kr) blocker dofetilide and the I(to)/IKur blockers AVE1231 and AVE0118 in five instrumented goats. Electrical remodeling progressively increased the AERP-prolonging effect of 3 mg/kg of AVE1231 and AVE0118 (2-fold increase in AERP at 72 hours vs SR, P < 0.01), but strongly decreased that of 10 mug/kg dofetilide (<0.5-fold, P < 0.05, at 300 and 400 ms basic cycle length). After 5 and 24 hours of tachypacing, the effect of 3 mg/kg amiodarone strongly increased (2-fold, P < 0.01 after 24 hours vs SR). This early gain in AERP prolongation was confirmed in anesthetized pigs with 3.5 hours of atrial tachypacing (2.4-fold increase, P < 0.01). At 72 hours of atrial tachypacing in the goat, however, the early gain was lost and the effect of amiodarone was similar again to that in SR. CONCLUSION: Atrial electrical remodeling changed the efficacy of the antiarrhythmic agents in a different way. The favorable efficacy profile of amiodarone during electrical remodeling, particularly the marked increase in AERP prolongation in early electrical remodeling, may explain its superior clinical efficacy over existing antiarrhythmic drugs. PMID- 17916156 TI - Differentiating atrioventricular nodal reentrant tachycardia from junctional tachycardia: novel application of the delta H-A interval. AB - INTRODUCTION: Junctional tachycardia (JT) and atrioventricular nodal reentrant tachycardia (AVNRT) can be difficult to differentiate. Yet, the two arrhythmias require distinct diagnostic and therapeutic approaches. We explored the utility of the delta H-A interval as a novel technique to differentiate these two tachycardias. METHODS: We included 35 patients undergoing electrophysiology study who had typical AVNRT, 31 of whom also had JT during slow pathway ablation, and four of whom had spontaneous JT during isoproterenol administration. We measured the H-A interval during tachycardia (H-A(T)) and during ventricular pacing (H A(P)) from the basal right ventricle. Interobserver and intraobserver reliability of measurements was assessed. Ventricular pacing was performed at approximately the same rate as tachycardia. The delta H-A interval was calculated as the H-A(P) minus the H-A(T). RESULTS: There was excellent interobserver and intraobserver agreement for measurement of the H-A interval. The average delta H-A interval was -10 ms during AVNRT and 9 ms during JT (P < 0.00001). For the diagnosis of JT, a delta H-A interval >or= 0 ms had the sensitivity of 89%, specificity of 83%, positive predictive value of 84%, and negative predictive value of 88%. The delta H-A interval was longer in men than in women with JT, but no gender-based differences were seen with AVNRT. There was no difference in the H-A interval based on age or =3.0 g/l. Global cognitive function was assessed serially with Mini-Mental State Examination (MMSE). The enrolled patients were followed for 2 years to observe if dementia was developed. There were 185 patients diagnosed as MCI, of which 17 (9.2%) deceased, 15 (8.1%) lost to follow-up, and 68 (36.8%) developed dementia during follow-up. Mean of MMSE score of the enrolled patients declined significantly during follow-up (22.0 +/- 3.0 vs. 18.1 +/- 5.8, p < 0.001). Patients with hyperfibrinogenaemia at baseline had greater MMSE decrement during follow-up than patients with normal fibrinogen level (-5.4 +/- 5.4 vs. -3.5 +/- 4.5, p < 0.05). Linear regression indicated that plasma fibrinogen level was associated with cognitive decline (R = 0.17, p < 0.05). Patients with hyperfibrinogenaemia had an increased risk for dementia and vascular dementia compared with patients with normal level of plasma fibrinogen (log rank test, p < 0.05). There was a trend that hyperfibrinogenaemia also increased risk for dementia of Alzheimer's type (p = 0.061). It can be concluded that plasma fibrinogen level may be associated with cognitive decline, and hyperfibrinogenaemia may increase risk for dementia in patients with MCI. PMID- 17916181 TI - Expression of Fas and Fas ligand in human testicular germ cell tumours. AB - In the present study, we analysed the expression of Fas ligand (FasL) and its cognate receptor Fas in 14 seminomatous testicular germ cell tumours (TGCT) and six normal testicular tissues obtained following orchiectomy. Tissue samples have been processed to prepare either total RNA or protein extracts or fixed and embedded in paraffin for immunohistochemistry (IHC) experiments. Quantitative RT PCR experiments demonstrated in TGCT a significant (p < 0.01) increase of the FasL mRNA expression of 21.1 +/- 5.4 fold, with respect to normal tissues. On the contrary, in the same cancer tissues, the levels of Fas mRNA were significantly (p < 0.01) reduced to 0.27 +/- 0.06 fold. These observations were confirmed in western blot experiments showing a significant increase of FasL and a concomitant decrease of Fas proteins in testicular cancer tissues, with respect to normal testis. Moreover, IHC experiments showed a strong FasL immuno-reactivity in six out of eight TGCT samples analysed, while Fas immuno-positivity was found in cancer cells of only two TGCT tissues. In addition, in all tumour samples, infiltrating lymphocytes were Fas positive. However, no correlation could be observed between Fas or FasL mRNA variations and clinical parameters such as patient's age, TNM stage or tumour size. We also compared the serum levels of soluble FasL (sFasL) of 15 patients affected by seminomatous TGCT, of four patients with non-seminomatous TGCT and six age-matched healthy males. No significant differences in sFasL serum level could be identified. In conclusion, our data demonstrated that the majority of seminomas are characterized by an increased expression of FasL and a concomitant reduction of Fas, with respect to human normal testis, and that sFasL serum level is not a tumour marker for patients affected by TGCT. PMID- 17916182 TI - CA repeat and RsaI polymorphisms in ERbeta gene are not associated with infertility in Indian men. AB - Oestrogen Receptor beta (ERbeta) gene plays an important role in the regulation of fertility in both males and females. Polymorphism in CA repeat located in the flanking region of ERbeta has been shown to be associated with several diseases, but its association with male infertility has not been analysed so far. However, RsaI polymorphism (rs1256049) in exon 5 of ERbeta has been shown to be associated with male infertility in Caucasian patients. Hence, we have analysed 695 Indian men, including 443 infertile and 252 fertile men to evaluate the association of CA repeat length and RsaI polymorphisms in male infertility. Our results revealed no significant difference in the distribution of CA repeat length between infertile (mean +/- SD 23.24 +/- 2.06, median 24) and fertile men (mean +/- SD 23.16 +/- 2.27, median 24). The analysis of dosage effect by classifying samples into SS (short/short), SL (short/long) and LL (long/long) groups also did not show any significant difference between infertile and fertile men. Similarly, RsaI polymorphism also did not show any significant difference between infertile and fertile men. Furthermore, the combined analysis of CA repeat and RsaI polymorphisms by haplotyping showed that the distribution of haplotypes was not significantly different between fertile and infertile men. Our results suggest that CA repeat length and RsaI polymorphisms in ERbeta are not associated with infertility in Indian men. PMID- 17916184 TI - Immunomagnetic separation of normal rat testicular cells from Roser's T-cell leukaemia cells is ineffective. AB - Elimination of contaminating malignant cells is crucial to avoiding cancer relapse in association with transplantation of autologous spermatogonial stem cells. In the clinical setting, there is presently no effective procedure for separating testicular cells from cancer cells. Here, CD4, a selective surface marker for Roser's rat leukaemic T-cells, was utilized to eliminate cancer cells from testicular cell samples from leukaemic piebald variegated (PVG) rats by magnetic-activated cell sorting (MACS). All animals receiving MACS-selected testicular cells died within 14-15 days. Only one-third of the contaminating leukaemic cells could be removed from testicular samples. An increase in antibody concentration enhanced the proportion of leukaemic cells removed from 27 to 49%. Variations in the cell size and expression of surface antigens on testicular leukaemic cells were the major obstacles to purification based on this marker. It is concluded that MACS does not prevent transmission of leukaemia to syngenic PVG rats when cells from leukaemic testes are used for testicular cell transplantation. PMID- 17916183 TI - Treatment of erectile dysfunction reduces psychological distress. AB - Mental stress is a risk factor for cardiovascular events in men with vascular risk factors (VRFs) and is also associated with erectile dysfunction (ED), a frequent complaint of men with VRFs. The aim of this study was to investigate the effect of inhibition of phosphodiesterase-5 or of placebo in men with ED and VRFs on self-evaluated psychological distress, erectile function and quality of sexual life. Thirty-six men with ED and VRFs were randomized to 4 weeks of tadalafil (20 mg/every other day) or placebo treatment. Sexual Health Inventory for Men (SHIM), questions 1-3 of Life Satisfaction (LiSat) questionnaire, Symptom Check-List-90R, a multidimensional inventory exploring psychological dimensions were applied before and after treatment. The SHIM score improved after treatment with tadalafil compared with baseline and with placebo (F = 10.38; p = 0.0030). Sexual life satisfaction (LiSat-2) was significantly improved after tadalafil and after placebo, but a strong positive correlation was observed between LiSat-2 and SHIM score after tadalafil treatment (r = 0.59, p = 0.0003) and not after placebo (r = 0.22, p = 0.189). Psychological features were significantly changed after treatment, although a specific effect of tadalafil vs. placebo was observed only for interpersonal sensitivity (F = 4.48; p = 0.042). Obsessive-compulsive dimension, depression, anxiety, psychoticism were significantly improved in the tadalafil group and in the placebo group, although the improvement was always more relevant after treatment with tadalafil. These preliminary data suggest that a short treatment of ED reduced psychological distress and improved quality of sexual life in men with VRFs. PMID- 17916185 TI - The impact of androgen receptor polymorphism and parental ethnicity on semen quality in young men from Latvia. AB - Recent studies on young men from the general population have demonstrated geographic and ethnic differences in semen quality. The aim of this study was to investigate whether reported ethnic differences in semen quality might be associated with the maternally derived CAG and GGN polymorphisms in the androgen receptor gene or paternal ethnicity. In total 114 military conscripts from Latvia were included in the study. Information on maternal and parental ethnicity was collected by questionnaires. CAG and GGN repeats were analysed by direct sequencing of leukocyte DNA. Men with Latvian mothers (n = 83) had marginally shorter CAG repeat length (21.6 +/- 2.9) as compared with those with non-Latvian mothers (22.9 +/- 3.2, n = 31), not reaching statistical significance (p = 0.053). Sperm concentration did not differ significantly between these two groups (76 +/- 59 and 70 +/- 52, p = 0.9 respectively). In contrast, significantly higher sperm concentration and total sperm count were found in men with Latvian fathers (n = 77) as compared with men with non-Latvian fathers (n = 37) (80 +/- 61 vs. 62 +/- 48, p = 0.035, for sperm concentration and 225.7 +/- 209 vs. 158.4 +/- 134.4, p = 0.002, for total sperm count respectively). CAG repeat length did not correlate with any semen parameters in the whole population. However, GGN repeat length correlated with semen volume: men with GGN > 23 presented with higher semen volume (3.2 +/- 2.1) as compared with men with GGN = 23 (2.6 +/- 1.3, p = 0.04) or GGN < 23 (2.0 +/- 1.2, p = 0.006). We conclude that GGN repeat length has an impact on semen volume, whereas differences in sperm numbers are associated with the paternal ethnicity. PMID- 17916186 TI - Misanalysis gave false association of mtDNA mutations with infertility. PMID- 17916187 TI - The isolation and characterization of temperature-dependent ricin A chain molecules in Saccharomyces cerevisiae. AB - Ricin is a heterodimeric plant protein that is potently toxic to mammalian cells. Toxicity results from the catalytic depurination of eukaryotic ribosomes by ricin toxin A chain (RTA) that follows toxin endocytosis to, and translocation across, the endoplasmic reticulum membrane. To ultimately identify proteins required for these later steps in the entry process, it will be useful to express the catalytic subunit within the endoplasmic reticulum of yeast cells in a manner that initially permits cell growth. A subsequent switch in conditions to provoke innate toxin action would permit only those strains containing defects in genes normally essential for toxin retro-translocation, refolding or degradation to survive. As a route to such a screen, several RTA mutants with reduced catalytic activity have previously been isolated. Here we report the use of Saccharomyces cerevisiae to isolate temperature-dependent mutants of endoplasmic reticulum targeted RTA. Two such toxin mutants with opposing phenotypes were isolated. One mutant RTA (RTAF108L/L151P) allowed the yeast cells that express it to grow at 37 degrees C, whereas the same cells did not grow at 23 degrees C. Both mutations were required for temperature-dependent growth. The second toxin mutant (RTAE177D) allowed cells to grow at 23 degrees C but not at 37 degrees C. Interestingly, RTAE177D has been previously reported to have reduced catalytic activity, but this is the first demonstration of a temperature-sensitive phenotype. To provide a more detailed characterization of these mutants we have investigated their N-glycosylation, stability, catalytic activity and, where appropriate, a three-dimensional structure. The potential utility of these mutants is discussed. PMID- 17916188 TI - Molecular basis for specificities of reactivating factors for adenosylcobalamin dependent diol and glycerol dehydratases. AB - Adenosylcobalamin-dependent diol and glycerol dehydratases are isofunctional enzymes and undergo mechanism-based inactivation by a physiological substrate glycerol during catalysis. Inactivated holoenzymes are reactivated by their own reactivating factors that mediate the ATP-dependent exchange of an enzyme-bound, damaged cofactor for free adenosylcobalamin through intermediary formation of apoenzyme. The reactivation takes place in two steps: (a) ADP-dependent cobalamin release and (b) ATP-dependent dissociation of the resulting apoenzyme reactivating factor complexes. The in vitro experiments with purified proteins indicated that diol dehydratase-reactivating factor (DDR) cross-reactivates the inactivated glycerol dehydratase, whereas glycerol dehydratase-reactivating factor (GDR) did not cross-reactivate the inactivated diol dehydratase. We investigated the molecular basis of their specificities in vitro by using purified preparations of cognate and noncognate enzymes and reactivating factors. DDR mediated the exchange of glycerol dehydratase-bound cyanocobalamin for free adeninylpentylcobalamin, whereas GDR cannot mediate the exchange of diol dehydratase-bound cyanocobalamin for free adeninylpentylcobalamin. As judged by denaturing PAGE, the glycerol dehydratase-DDR complex was cross-formed, although the diol dehydratase-GDR complex was not formed. There were no specificities of reactivating factors in the ATP-dependent dissociation of enzyme-reactivating factor complexes. Thus, it is very likely that the specificities of reactivating factors are determined by the capability of reactivating factors to form complexes with apoenzymes. A modeling study based on the crystal structures of enzymes and reactivating factors also suggested why DDR cross-forms a complex with glycerol dehydratase, and why GDR does not cross-form a complex with diol dehydratase. PMID- 17916189 TI - Identification of calreticulin as a ligand of GABARAP by phage display screening of a peptide library. AB - 4-Aminobutyrate type A (GABA(A)) receptor-associated protein (GABARAP) is a ubiquitin-like modifier implicated in the intracellular trafficking of GABA(A) receptors, and belongs to a family of proteins involved in intracellular vesicular transport processes, such as autophagy and intra-Golgi transport. In this article, it is demonstrated that calreticulin is a high affinity ligand of GABARAP. Calreticulin, although best known for its functions as a Ca(2+) dependent chaperone and a Ca(2+) -buffering protein in the endoplasmic reticulum, is also localized to the cytosol and exerts a variety of extra-endoplasmic reticulum functions. By phage display screening of a randomized peptide library, peptides that specifically bind GABARAP were identified. Their amino acid sequences allowed us to identify calreticulin as a potential GABARAP binding protein. GABARAP binding to calreticulin was confirmed by pull-down experiments with brain lysate and colocalization studies in N2a cells. Calreticulin and GABARAP interact with a dissociation constant K(d) = 64 nm and a mean lifetime of the complex of 20 min. Thus, the interaction between GABARAP and calreticulin is the strongest so far reported for each protein. PMID- 17916190 TI - Dual effect of echinomycin on hypoxia-inducible factor-1 activity under normoxic and hypoxic conditions. AB - Hypoxia-inducible factor-1 (HIF-1) is now recognized as a possible target for cancer treatment. This transcription factor is responsible for the overexpression of several genes favouring cancer cell survival and inducing neo-angiogenesis. Echinomycin has recently been described to inhibit HIF-1 DNA binding and transcriptional activity. In this work, it is shown that echinomycin strongly inhibits the activity of HIF-1 under hypoxic conditions, and also interferes with the activity of other transcription factors. These results demonstrate the lack of specificity of this molecule. Moreover, it is demonstrated that echinomycin induces an increase in HIF-1 activity under normoxic conditions, parallel to an increase in the expression of HIF-1 target genes. This effect is caused by an increase in HIF-1alpha protein level, resulting from an increase in the transcription of the HIF-1A gene in the presence of a low concentration of echinomycin. Transfection experiments with HIF-1alpha promoter constructs revealed the presence of an Sp1 binding element responsive to echinomycin. Furthermore, echinomycin enhanced Sp1 activity, as measured by the use of a specific reporter system. These findings show, for the first time, that echinomycin has a dual effect on HIF-1 activity under normoxic and hypoxic conditions, demonstrating that this molecule cannot be used in cancer treatment. PMID- 17916191 TI - The impact of the regulatory design on the response of epidermal growth factor receptor-mediated signal transduction towards oncogenic mutations. AB - Epidermal growth factor receptor (EGFR)-mediated signal transduction is often hyperactivated in tumour cells and therefore considered a promising target for cancer therapy. A number of computational models have been developed which describe the pathway in great detail. These models are similar in their description of the activation events. The deactivation of the EGFR signalling seems to be cell type-specific and is less understood. Deactivation via receptor internalization, feedback inhibition of son of sevenless (SOS) by double phosphorylated, extracellular signal-regulated kinase (ERKPP) or transiently activated Ras-GTPase activating protein (Ras-GAP) proteins is discussed to play a role. In this study we address the question of to what extent the effect of oncogenic perturbations on EGFR signalling depend on the specific regulation structure. This is investigated using a detailed pathway model under two regulatory modes: the negative feedback via ERKPP to SOS and feed-forward deactivation via transiently activated Ras-GAP proteins. We show that the effect of receptor overexpression differs qualitatively under both regulations. In the system with transiently activated Ras-GAP it may result in an attenuation of the ERK activation. Such a nonintuitive effect was also observed experimentally. In general we find the model with transiently activated Ras-GAP to have a higher robustness towards receptor overexpression and Ras mutations. In particular, we demonstrate that this model can compensate for these oncogenic perturbations if the regulation is strong. The negative feedback can not protect the system against Ras mutations. A general sensitivity analysis, however, shows a higher robustness of the model under negative feedback, indicating the limited significance of such analyses for the prediction of specific oncogenic perturbations. PMID- 17916192 TI - Respiratory involvement in toxic epidermal necrolysis portends a poor prognosis that may not be reflected in SCORTEN. PMID- 17916193 TI - Basal cell carcinomas of the inner canthus: incidence of incomplete excision according to topographical localization of tumours. PMID- 17916194 TI - Transient macular erythema with eosinophilia in a patient carrying the FIP1L1 PDGFRA fusion gene. PMID- 17916195 TI - Recurrent KIND1 (C20orf42) gene mutation, c.676insC, in a Brazilian pedigree with Kindler syndrome. PMID- 17916196 TI - Treatment of pyoderma gangrenosum with intravenous immunoglobulin. AB - BACKGROUND: Intravenous immunoglobulin (IVIG) is increasingly being used to treat inflammatory and autoimmune disease. OBJECTIVES: To elucidate the efficacy of IVIG as an adjunct treatment for pyoderma gangrenosum (PG). PATIENTS/METHODS: Ten patients with PG were treated with IVIG at Johns Hopkins Department of Dermatology. All patients had severe mutilating and/or refractory disease requiring multi-agent therapy. The charts were reviewed retrospectively. RESULTS: Seven of the ten patients had clearance of PG lesions in the setting of IVIG and six of these patients maintained efficacy with repeated IVIG treatment. Five patients complained of nausea with treatment, and in one case nausea was severe and intractable. One patient developed an immune reaction requiring diphenhydramine and methylprednisolone and another experienced aseptic meningitis. CONCLUSIONS: IVIG may be an effective adjuvant in the treatment of PG and has an acceptable side-effect profile. Randomized, placebo-controlled, double blinded trials are needed to confirm this hypothesis. PMID- 17916198 TI - Adalimumab for treatment of pyoderma gangrenosum. PMID- 17916197 TI - Environmental factors, parental atopy and atopic eczema in primary-school children: a cross-sectional study in Taiwan. AB - BACKGROUND: Parental atopy and environmental exposure are recognized risk factors for atopic eczema (AE) in childhood. However, the relative contributions of specific risk factors and the overall contributions of hereditary and environmental exposure remain unexplored. OBJECTIVES: To identify risk factors, estimate the population attributable risk (PAR) of environmental exposure, and compare the AE data for boys vs. girls in primary-school children. METHODS: During a February to June 2001 cross-sectional, Taiwan-based questionnaire survey, we investigated 23 980 children from 22 primary schools, all located within 1 km of an air-monitoring station. RESULTS: The 12-month prevalence of AE was reported as 6.1% in boys and 4.9% in girls. In both sexes, the risk of AE was strongly associated with parental atopy and perceived ambient air pollution. The presence of cockroaches [odds ratio (OR) 1.18, 95% confidence interval (CI) 1.00 1.40] and visible mould on walls at home (OR 1.46, 95% CI 1.22-1.70) were also significantly related to AE for girls; however, only visible mould on walls (and not the presence of cockroaches) at home was related to AE for boys (OR 1.40, 95% CI 1.18-1.66). While mutually adjusted models were applied, we found adjusted ORs and PARs were similar in boys and girls in hereditary and outdoor environmental factors. The PAR of indoor environmental factors was higher in girls (8.4%) than in boys (5.5%). There was no interaction between parental atopy and environmental factors. CONCLUSIONS: Parental atopy contributed more to AE than indoor or outdoor environmental factors. Girls may be more susceptible to indoor environmental factors than boys. PMID- 17916199 TI - A sporadic case of early-onset sarcoidosis resembling Blau syndrome due to the recurrent R334W missense mutation on the NOD2 gene. AB - Sarcoidosis is a multisystem granulomatous disorder characterized by the infiltration of noncaseating granulomata in the affected tissues. We report here the clinical case of a Caucasian Spanish patient suffering from sporadic early onset sarcoidosis (EOS) with simultaneous cutaneous and articular symptoms. NOD2 (nucleotide-binding oligomerization domain; previously known as CARD15, caspase recruitment domain) gene mutational analysis revealed the presence of the recurrent R334W missense mutation. As in previously reported EOS cases, our patient was initially misdiagnosed with dermatitis. PMID- 17916200 TI - Melanocortin 1 receptor (MC1R) genotype influences erythemal sensitivity to psoralen-ultraviolet A photochemotherapy. AB - BACKGROUND: The melanocortin 1 receptor (MC1R) is a highly polymorphic G protein coupled receptor. Inheritance of various MC1R alleles has been associated with a red hair/fair skin phenotype, increased incidence of skin cancer and altered sensitivity to ultraviolet (UV) radiation. OBJECTIVES: To investigate whether MC1R genotype influences erythemal sensitivity to psoralen-UVA photochemotherapy (PUVA) in patients with psoriasis and other common skin diseases. METHODS: Patients (n = 111) about to start PUVA were recruited to the study. Erythemal responses were assessed visually at 72 h and 96 h following PUVA by assessment of the minimal phototoxic dose (MPD). MC1R genotype was determined by direct sequencing. RESULTS: Inheritance of the MC1R Arg(151)Cys allele was associated with a red hair phenotype (odds ratio 25.19, P = 0.0004). In contrast, inheritance of the Val(60)Leu and Arg(163)Gln SNPs was associated with increased PUVA erythemal sensitivity (reduced MPD) 72 h following treatment in all patients (n = 111; Val(60)Leu chi(2) = 5.764, P = 0.016; Arg(163)Gln chi(2) = 5.469, P = 0.019) and in a subset of patients with psoriasis (n = 55; Val(60)Leu chi(2) = 4.534, P = 0.033; Arg(163)Gln chi(2) = 7.298, P = 0.007). Inheritance of two or more MC1R SNPs was also associated with increased PUVA erythemal sensitivity (reduced MPD) in both patient groups (n = 111; chi(2) = 8.166, P = 0.017; n = 55; chi(2) = 10.303, P = 0.016). CONCLUSIONS: Our data demonstrate that MC1R genotype influences PUVA erythemal sensitivity in patients with psoriasis and other common skin diseases. PMID- 17916201 TI - Herlitz junctional epidermolysis bullosa: laminin-5 mutational profile and carrier frequency in the Italian population. AB - BACKGROUND: Herlitz junctional epidermolysis bullosa (HJEB; MIM 226700) is a rare epithelial adhesion disorder caused by null mutations in any of the three genes encoding the alpha3, beta3 and gamma2 chains of laminin-5, and is mainly characterized by extensive mucocutaneous blistering, recurrent infections and early lethality. OBJECTIVES: To perform immunoepitope mapping, electron microscopy and molecular analysis of five Italian patients with HJEB in order to complete the clinical and molecular characterization of patients with HJEB collected in the Italian Registry of hereditary epidermolysis bullosa (IRHEB) and to calculate the HJEB carrier frequency in this population. METHODS: Skin biopsies from perilesional skin of all patients were employed for immunoepitope mapping and electron microscopy examination. Blood genomic DNA was used for mutation analysis in the LAMA3, LAMB3 and LAMC2 genes by heteroduplex scanning, preceded by a search for Italian recurrent mutations. Carrier frequency calculation was performed assuming Hardy-Weinberg equilibrium. RESULTS: Two novel mutations in the LAMA3 (p.R782X) and LAMC2 (c.3235delA) genes, as well as three known and recurrent mutations in the LAMB3 (c.31insC and p.R81X) and LAMC2 (p.Y355X) genes were identified. Based on disease incidence reported in the IRHEB and the prevalence of mutations in each laminin-5 gene, the population carrier risk for HJEB was calculated to be one in 375. CONCLUSIONS: Our delineation of a laminin-5 mutational spectrum in the general Italian population provides a solid basis for expedited diagnosis, accurate genetic counselling and DNA-based prenatal testing for Italian families at risk for recurrence of HJEB. PMID- 17916202 TI - Understanding spitzoid tumours: new insights from molecular pathology. AB - Spitzoid tumours are a morphologically diverse group of lesions that share histological similarity to the Spitz naevus, a benign melanocytic skin tumour. Distinguishing classic Spitz naevi from cutaneous malignant melanoma is usually achievable on standard histology sections, but occasionally equivocal lesions are encountered that show features intermediate between these two entities and consequently generate considerable clinical and histopathological concern. The nomenclature and diagnostic criteria for spitzoid lesions are not standardized and this article begins by considering the adverse effect this has on our understanding of spitzoid tumour biology. Investigations of some of the hallmark features of cancer and neoplasia in spitzoid tumours are described, and the contribution of these studies to our understanding of spitzoid tumour biology is considered, along with their potential diagnostic utility. These studies compare spitzoid tumours with better-characterized melanocytic lesions, and from such comparisons assumptions concerning the biological nature of different spitzoid tumours can be made. In contrast, investigations of the mitogen-activated protein kinase (MAPK) pathway and DNA gains and losses have suggested that Spitz naevi may be genetically distinct from other melanocytic tumours. The studies that led to this conclusion are reviewed, as well as subsequent work examining whether the same applies to all spitzoid tumours. Possible explanations for the considerable inconsistencies within some of these data are explored. Finally, potential pathways of tumour progression within spitzoid lesions are considered, with an emphasis placed upon insights gained from investigations of MAPK genes and DNA gains and losses. PMID- 17916203 TI - A novel homozygous mutation of the EVER1/TMC6 gene in a Japanese patient with epidermodysplasia verruciformis. PMID- 17916204 TI - Effects of etanercept therapy on fatigue and symptoms of depression in subjects treated for moderate to severe plaque psoriasis for up to 96 weeks. PMID- 17916205 TI - Cutaneous Langerhans cell histiocytosis in an elderly man successfully treated with narrowband ultraviolet B. PMID- 17916207 TI - Pyoderma gangrenosum and interleukin 8. PMID- 17916206 TI - Reduction of immunosuppression for transplant-associated skin cancer: thresholds and risks. AB - BACKGROUND: Although evidence supports the efficacy of reducing immunosuppression for transplant-associated skin cancer, clinical thresholds for and risks associated with reduction are not well defined. OBJECTIVES: In this study, experienced transplant physicians were surveyed regarding appropriate thresholds for consideration of reduction of immunosuppression and the likelihood of rejection and allograft compromise associated with various levels of reduction. PATIENTS AND METHODS: Fifty-two transplant physicians reviewed 13 hypothetical patient scenarios with graduated morbidity and mortality risk and provided opinions on the degree of reduction of immunosuppression that was warranted and the risks associated with various degrees of reduction. RESULTS: Renal, liver and cardiac transplant physicians generally concurred on the level of reduction of immunosuppression warranted by various degrees of skin cancer. As morbidity and mortality from skin cancer increased, physicians were more likely to accept risk to allograft function from more aggressive reduction. CONCLUSIONS: Reduction of immunosuppression is considered a reasonable adjuvant strategy in recipients of solid organ transplants who have substantial morbidity and mortality risk from skin cancer. Physicians are willing to accept an increased risk of allograft compromise when confronted by severe or extensive skin cancer. Further research is needed to define the precise correlation among levels of reduction of immunosuppression, therapeutic efficacy, and concomitant risks. PMID- 17916208 TI - CTACK /CCL27 expression in psoriatic skin and its modification after administration of etanercept. AB - BACKGROUND: Tumour necrosis factor-alpha upregulates the expression of a cutaneous T cell-attracting chemokine (CTACK/CCL27), that promotes migration of cutaneous lymphocyte-associated antigen-positive lymphocytes into the skin. The role of CTACK/CCL27 in pathogenesis of psoriasis has recently been documented but no data are available at the present time on its modification in psoriatic cutaneous tissue after administration of etanercept. OBJECTIVES: To evaluate modifications of CTACK/CCL27 expression in skin of patients with psoriasis after administration of etanercept and their relation with disease activity. METHODS: Twenty-two patients with moderate to severe psoriasis underwent clinical, histological and immunohistochemical evaluations of disease activity at baseline and at 12 and 24 weeks after starting treatment with etanercept. RESULTS: All selected patients experienced an improvement of Psoriasis Area and Severity Index (PASI) score (P < 0.001) and Dermatology Life Quality Index score (P < 0.001) during the treatment. Skin histological abnormalities showed statistically significant modifications during treatment (P < 0.001). Immunohistochemical expression of CTACK/CCL27 decreased significantly (P < 0.001) and its relation with final PASI score was statistically significant (P < 0.05); the pattern of distribution of CTACK/CCL27 immunoreactivity significantly moved from diffuse and predominantly suprabasal to basal (P < 0.001) and the restoration of basal distribution of CTACK/CCL27 was also significantly related to clinical improvement of cutaneous disease (P < 0.001). CONCLUSIONS: Etanercept induces a clinical and histological improvement of psoriatic disease, promoting a reduction in CTACK/CCL27 cutaneous immunostaining and favouring the restoration of physiological CTACK/CCL27 epidermal expression. Moreover, CTACK/CCL27 reduction in cutaneous expression during administration of etanercept could be considered a favourable prognostic marker. PMID- 17916209 TI - A case of primary anetoderma in an infant. PMID- 17916210 TI - Potassium channel openers accelerate epidermal barrier recovery. AB - BACKGROUND: Maintenance of a competent permeability barrier in the face of external and internal stressors requires signals between the stratum corneum interface and the metabolic machinery in the underlying nucleated layers. For example, reductions in the ion gradients for Ca2+ after acute barrier disruption stimulate lamellar body (LB) secretion, a response required to restore barrier homeostasis. Although alterations in external K+ levels also regulate barrier recovery after acute insults, the mechanisms whereby K+ regulates barrier function remain unknown. OBJECTIVES: To evaluate effects of regulators of K+ channels on barrier homeostasis in hairless mice. METHODS: We tested a number of chemically different drugs that alter intracellular K+ levels. Results Single applications of either K+ channel openers (i.e. 1-EBIO, minoxidil, diazoxide) or the K+ ionophore, valinomycin, accelerated barrier recovery after acute insults to murine skin, paralleled by a reduction in intracellular K+ levels in cultured human keratinocytes. In contrast, applications of K+ channel blockers (i.e. gilbenclamide, dequalinium) delayed barrier recovery. Alterations in intracellular K+ regulated barrier homeostasis by either stimulating (reduced K+) or inhibiting (elevated K+) LB secretion. Finally, development of epidermal hyperplasia, a downstream consequence of barrier disruption, was also inhibited by agents that reduce intracellular K+ levels. CONCLUSIONS: These results demonstrate that changes in K+ levels that can be presumed to occur after barrier disruption signal metabolic responses, i.e. LB secretion, which accelerates normalization of barrier function. PMID- 17916211 TI - Prevalence of Staphylococcus aureus toxins and nasal carriage in furuncles and impetigo. AB - BACKGROUND: The precise role of Staphylococcus aureus toxins and nasal carriage in common skin infections remains unclear. OBJECTIVES: To seek correlations between toxin expression, S. aureus nasal carriage and clinical manifestations in patients with community-acquired furuncles and impetigo. METHODS: From November 2004 to August 2005, we studied clinical data and bacteriological samples prospectively collected from 121 patients presenting with furuncles or impetigo. RESULTS: Sixty-four patients (31 with furuncles and 33 with impetigo) had S. aureus-positive skin culture. Panton-Valentine leukocidin (PVL) genes were present in 13 of 31 (42%) isolates from furuncles and were associated with epidemic furunculosis. Exfoliative toxin genes were present in 10 of 10 (100%) and 12 of 21 (57%) bullous and nonbullous impetigo isolates, respectively. Nasal carriage of S. aureus was found in 58% of patients overall. It was strongly associated with chronic furunculosis but not with simple furuncles (88% vs. 29%, P < 0.007). Skin and nose isolates from a given patient always had identical characteristics. Methicillin-resistant S. aureus accounted for four of 64 (6%) positive skin cultures. CONCLUSIONS: PVL is not involved in all types of furuncles but is associated with epidemic furunculosis. Both bullous and nonbullous forms of impetigo are associated with exfoliative toxins. Staphylococcus aureus nasal carriage is associated with the chronicity of furuncles. PMID- 17916212 TI - Cathelicidin LL-37 induces the generation of reactive oxygen species and release of human alpha-defensins from neutrophils. AB - BACKGROUND: Psoriasis is characterized by epidermal infiltration of neutrophils that destroy invading microorganisms via a potent antimicrobial arsenal of oxidants and antimicrobial agents. In contrast to atopic dermatitis, psoriasis exhibits low levels of skin infections due to the presence of antimicrobial agents, including cathelicidin LL-37. LL-37 kills a broad spectrum of microbes, and activates neutrophil chemotaxis. OBJECTIVE: To determine whether or not LL-37 could regulate additional neutrophil functions such as production of cytokines/chemokines, reactive oxygen species and release of neutrophil antimicrobial peptides. METHODS: Human peripheral blood neutrophils were used in this study. The production of interleukin (IL)-8 and release of alpha-defensins were analysed by enzyme-linked immunosorbent assay, and real-time polymerase chain reaction (PCR) was used to quantify alpha-defensin gene expression. Phosphorylation of mitogen-activated protein kinase (MAPK) was determined by Western blotting. The generation of reactive oxygen species was examined using flow cytometry, and intracellular Ca(2+) mobilization was measured using a calcium assay kit. RESULTS: LL-37 enhanced the production of IL-8 under the control of MAPK p38 and extracellular signal regulated kinase (ERK), as evidenced by the inhibitory effects of p38 and ERK1/2 inhibitors on LL-37-mediated IL-8 production. Furthermore, LL-37 induced phosphorylation of p38 and ERK. We also revealed that LL-37 stimulated the generation of reactive oxygen species dose- and time-dependently, most probably via NADPH oxidase activation and intracellular Ca(2+) mobilization. Finally, LL-37 induced both mRNA expression and protein release of alpha-defensins, known as human neutrophil peptide 1-3. CONCLUSION: Taken together, we suggest that in addition to its microbicidal properties, LL-37 may contribute to innate immunity by enhancing neutrophil host defence functions at inflammation and/or infection sites. PMID- 17916213 TI - Increased nuclear beta-catenin in suprabasal involved psoriatic epidermis. AB - BACKGROUND: Psoriasis is a common inflammatory skin disease characterized by abnormal keratinocyte proliferation and differentiation, increased angiogenesis and inflammation. There is evidence that some keratinocyte differentiation events are controlled by changes in cell-cell adhesion. beta-catenin is a 94-kDa protein which has a dual function as a component of intercellular adherens junctions and also as a transcription factor as part of the Wnt signalling pathway. beta catenin is not required for keratinocyte proliferation but has been shown to regulate keratinocyte stem cells and hair follicle morphogenesis. OBJECTIVES: To investigate the distribution and function of beta-catenin in involved psoriatic epidermis and in epidermal keratinocytes. METHODS: Biopsies were obtained from patients with psoriasis and from normal controls. The distribution of beta catenin was investigated using antibodies to both total and unphosphorylated active beta-catenin. Luciferase assays were used to measure transcriptional activation of transglutaminase 1 (TGase 1) and involucrin and to investigate the functional role of beta-catenin in interfollicular keratinocytes. RESULTS: Increased nuclear beta-catenin was seen in lesional suprabasal psoriatic epidermis compared with uninvolved or normal skin. Increased active unphosphorylated beta-catenin was also detected within the differentiating compartment of involved psoriatic epidermis. Expression of TGase 1 overlapped with beta-catenin in suprabasal lesional psoriasis. The TGase 1 promoter was positively regulated by activated beta-catenin and by the glycogen synthase kinase binding protein, suggesting that beta-catenin and glycogen synthase kinase 3beta may regulate TGase 1 expression. CONCLUSIONS: This is the first report to convincingly demonstrate increased beta-catenin in involved psoriasis and to implicate beta-catenin in the regulation of TGase 1. This evidence suggests a role for beta-catenin signalling in regulating keratinocyte differentiation in interfollicular skin in addition to previously reported functions in stem cell fate determination, hair follicle regulation and skin tumorigenesis. PMID- 17916214 TI - Treatment of xanthelasma palpebrarum by 1064-nm Q-switched Nd:YAG laser: a study of 11 cases. AB - BACKGROUND: Xanthelasma palpebrarum (xanthelasma) is a common disease involving the eyelids. It consists of yellowish plaques of esterified cholesterol situated in the middle and superficial layers of the dermis. Many techniques, including laser treatments, have been proposed to destroy these lesions. A preliminary study demonstrated the efficacy of 1064-nm Q-switched Nd:YAG laser in treating xanthelasma. OBJECTIVES: To present the results of a prospective study using 1064 nm Q-switched Nd:YAG laser to treat xanthelasma. METHODS: Thirty-eight lesions in 11 patients were treated. RESULTS: The results observed after a single treatment by Q-switched Nd:YAG laser were scored as good or excellent in eight patients, i.e. for 26 of 38 lesions. CONCLUSIONS: The treatment of xanthelasma by 1064-nm Q switched Nd:YAG laser is a valuable treatment option to eliminate lesions, with rapid and good quality healing. The absence of any associated skin destruction allows treatment to be repeated when necessary. PMID- 17916215 TI - Cutaneous Malassezia flora in atopic dermatitis differs between adults and children. AB - BACKGROUND: Malassezia species are suspected to be involved in the development of skin lesions in atopic dermatitis (AD) when the response of adult AD to anti inflammatory treatments is poor. However, a comparative analysis of Malassezia flora between adults and children with AD has not been performed. OBJECTIVES: To compare the cutaneous Malassezia flora between adults and children with AD. METHODS: Scale samples were collected from skin lesions of 58 patients with AD in the head and neck regions (28 males and 30 females; 31 children and 27 adults), and fungal DNA was extracted from the samples directly. The number and identities of the Malassezia species were analysed with high accuracy using a polymerase chain reaction-based culture-independent method. The in vivo level of anti Malassezia IgE antibody was also assayed. RESULTS: Malassezia restricta was the predominant species in the children with AD, while both M. restricta and M. globosa predominated in the adults. The adults showed increased sensitization in terms of anti-Malassezia-specific IgE responses in the sera to both M. globosa and M. restricta in comparison with the children. CONCLUSIONS: The cutaneous Malassezia flora differs significantly between the two age groups. PMID- 17916216 TI - COL7A1 mutational analysis in Korean patients with dystrophic epidermolysis bullosa. PMID- 17916217 TI - Psoriasis patients show signs of insulin resistance. AB - BACKGROUND: Recent observations suggest that psoriasis is a risk factor for the development of coronary artery calcification which in turn represents an indicator for atherosclerosis. OBJECTIVE: To clarify a possible pathogenetic link between psoriasis and atherosclerosis, we studied the metabolic state of patients with psoriasis. METHODS: Thirty-nine consecutive patients with moderate-to-severe plaque-type psoriasis were enrolled in the study. Detailed information was obtained on the patients' clinical picture and history of psoriasis, smoking habits and medication. The body mass index (BMI) of the patients was calculated. Laboratory investigations focused on values for inflammation, lipid profile and multiple cytokines. The intima-media thickness of the carotid artery was measured by ultrasound, and an oral glucose tolerance test was performed to calculate the homeostasis model assessment of insulin resistance (HOMA). RESULTS: Numerous well recognized correlations such as between BMI and HOMA (P < 0.02) as well as BMI and vessel wall thickness (P < 0.05) were successfully reproduced, thus confirming consistency of our dataset. With regard to psoriasis, we observed a significant correlation between the Psoriasis Area and Severity Index (PASI) score and insulin secretion. Moreover, the PASI score was significantly correlated with serum resistin levels--a cytokine known to be increased in insulin resistance. CONCLUSIONS: Taken together, several measurements indicative of insulin resistance were found to be significantly correlated with the PASI score. The concept of insulin resistance as a consequence of chronic inflammation and possible pathogenetic cause for comorbidities known to be associated with psoriasis is supported by these data. Our findings validate further studies on larger cohorts as well as interventional studies. PMID- 17916218 TI - Late-onset neutropenia following rituximab treatment in patients with autoimmune diseases. PMID- 17916219 TI - Hypertriglyceridaemia during treatment with adalimumab in psoriatic arthritis. PMID- 17916220 TI - Recovery from Sezary syndrome following Mycobacterium avium spondylitis. PMID- 17916221 TI - Successful treatment of severe psoriatic natal cleft fissuring with tissue adhesive. PMID- 17916222 TI - Does early onset of non-medical use of prescription drugs predict subsequent prescription drug abuse and dependence? Results from a national study. AB - AIMS: The present study examined the associations between early onset of non medical use of prescription drugs (NMUPD) (i.e. sedatives, tranquilizers, opioids, stimulants) and the development of prescription drug abuse and dependence in the United States. DESIGN: Data were collected from structured diagnostic interviews using the National Institute on Alcohol Abuse and Alcoholism (NIAAA) Alcohol Use Disorder and Associated Disabilities Interview Schedule: Diagnostic and Statistical Manual version IV (DSM-IV). SETTING: National prevalence estimates were derived from the 2001-2002 National Epidemiologic Survey on Alcohol and Related Conditions (NESARC, n = 43,093). PARTICIPANTS: A nationally representative cross-sectional sample of civilian non institutionalized adults aged 18 years or older in the United States, of whom 52% were women, 71% white, 12% Hispanic, 11% African American, 4% Asian and 2% Native American or of other racial background. FINDINGS: A higher percentage of individuals who began using prescription drugs non-medically at or before 13 years of age were found to have developed prescription drug abuse and dependence versus those individuals who began using at or after 21 years of age. Multivariate logistic regression analyses indicated that the odds of developing any life-time prescription drug abuse among non-medical users was reduced by approximately 5% with each year non-medical use was delayed [adjusted odds ratio (AOR) = 0.95, 95% CI = 0.94, 0.97], and that the odds of developing any life-time prescription drug dependence were reduced by about 2% with each year onset was delayed (AOR = 0.98, 95% CI = 0.96, 1.00) when controlling for relevant covariates. CONCLUSIONS: The results of this study indicate that early onset of NMUPD was a significant predictor of prescription drug abuse and dependence. These findings reinforce the importance of developing prevention efforts to reduce NMUPD and diversion of prescription drugs among children and adolescents. PMID- 17916223 TI - Relationship between menthol cigarettes and smoking cessation among African American light smokers. AB - AIMS: To determine whether African American light smokers who smoked menthol cigarettes had lower cessation when treated with nicotine replacement therapy and counseling. DESIGN: Data were derived from a clinical trial that assessed the efficacy of 2 mg nicotine gum (versus placebo) and counseling (motivational interviewing counseling versus Health Education) for smoking cessation among African American light smokers (smoked < or = 10 cigarettes per day). PARTICIPANTS: The sample consisted of 755 African American light smokers. MEASUREMENTS: The primary outcome variable was verified 7-day point-prevalence smoking cessation at 26 weeks follow-up. Verification was by salivary cotinine. FINDINGS: Compared to non-menthol smokers, menthol smokers were younger and less confident to quit smoking (P = 0.023). At 26 weeks post-randomization, 7-day verified abstinence rate was significantly lower for menthol smokers (11.2% versus 18.8% for non-menthol, P = 0.015). CONCLUSIONS: Among African American light smokers, use of menthol cigarettes is associated with lower smoking cessation rates. Because the majority of African American smokers use menthol cigarettes, a better understanding of the mechanism for this lower quit rate is needed. PMID- 17916224 TI - Developing limits for driving under cannabis. AB - OBJECTIVE: Development of a rational and enforceable basis for controlling the impact of cannabis use on traffic safety. METHODS: An international working group of experts on issues related to drug use and traffic safety evaluated evidence from experimental and epidemiological research and discussed potential approaches to developing per se limits for cannabis. RESULTS: In analogy to alcohol, finite (non-zero) per se limits for delta-9-tetrahydrocannabinol (THC) in blood appear to be the most effective approach to separating drivers who are impaired by cannabis use from those who are no longer under the influence. Limited epidemiological studies indicate that serum concentrations of THC below 10 ng/ml are not associated with an elevated accident risk. A comparison of meta-analyses of experimental studies on the impairment of driving-relevant skills by alcohol or cannabis suggests that a THC concentration in the serum of 7-10 ng/ml is correlated with an impairment comparable to that caused by a blood alcohol concentration (BAC) of 0.05%. Thus, a suitable numerical limit for THC in serum may fall in that range. CONCLUSIONS: This analysis offers an empirical basis for a per se limit for THC that allows identification of drivers impaired by cannabis. The limited epidemiological data render this limit preliminary. PMID- 17916225 TI - Melatonin for perceived sleep disturbances associated with benzodiazepine withdrawal among patients in methadone maintenance treatment: a double-blind randomized clinical trial. AB - AIMS: To evaluate the effectiveness of melatonin in attenuating sleep difficulties during benzodiazepine (BDZ) withdrawal. DESIGN: Double-blind cross over control study. SETTING: Methadone maintenance treatment clinic. PARTICIPANTS: Eighty patients enrolled at a community methadone maintenance clinic recruited to a BDZ withdrawal programme. INTERVENTION: Melatonin (5 mg/day) or placebo: 6 weeks one arm, 1 week washout, 6 weeks other arm. MEASUREMENTS: Urine BDZ; self-reported Pittsburgh Sleep Quality Index (PSQI) and the Center for Epidemiologic Studies Depression (CES-D) questionnaires administered at baseline, and at 6, 7 and 13 weeks. FINDINGS: Sixty-one patients (77.5% in the 'melatonin first' condition and 75% in the 'placebo first' condition) completed 6 weeks of treatment, showing a similar BDZ discontinuation rate of 11/31 and 11/30, respectively. PSQI scores were significantly lower (indicating better sleep quality) in the 22 patients who discontinued BDZ (8.9 +/ 0.9) than in 39 with urine BDZ (11.2 +/- 0.7, P = 0.04). Sleep quality in patients who continued abusing BDZ improved more in the 'melatonin first' group than in the 'placebo first' group, with no differences in sleep quality improvement in patients who stopped BDZ. CONCLUSION Most improvement in sleep quality was attributed to BDZ discontinuation. Although melatonin did not enhance BDZ discontinuation, it improved sleep quality, especially in patients who did not stop BDZ. PMID- 17916226 TI - Cytosol to lysosome transport of intracellular antigens during immune surveillance. AB - The delivery of intracellular substrates such as misfolded proteins and damaged organelles from the cytosol to the lysosome for degradation is crucial for cell survival. Multiple transport pathways including bulk autophagy (microautophagy and macroautophagy) and chaperone-mediated autophagy (CMA) have been identified to efficiently facilitate this transit of macromolecules from the cytoplasm to acidic vacuolar organelles. While autophagy plays a role in the general housekeeping of cells, it also functions in more specialized processes such as development and differentiation, responses to physiological stress and immunity. The presentation of both exogenous and endogenous antigens (Ag) by major histocompatibility complex (MHC) class II molecules to CD4(+) T lymphocytes is critical for the induction of tolerance to self Ag as well as the development of immunity against intracellular pathogens and tumors. Here, we discuss the class II-mediated presentation of several endogenous Ag, dependent on either macroautophagy or CMA for their transport from the cytosol to endosomal/lysosomal compartments. Thus, the various pathways of autophagy as routes of cytoplasmic Ag delivery to lysosomes have significant implications for the MHC class II-mediated immune response to intracellular pathogens and cancer. PMID- 17916228 TI - Inflammation triggers colony forming endothelial cell mobilization after angioplasty in chronic lower limb ischemia. PMID- 17916227 TI - Dominant-negative behavior of mammalian Vps35 in yeast requires a conserved PRLYL motif involved in retromer assembly. AB - The retromer protein complex assists in recycling selected integral membrane proteins from endosomes to the trans Golgi network. One protein subcomplex (Vps35p, Vps26p and Vps29p) combines with a second (Vps17p and Vps5p) to form a coat involved in sorting and budding of endosomal vesicles. Yeast Vps35p (yVps35) exhibits similarity to human Vps35 (hVps35), especially in a completely conserved PRLYL motif contained within an amino-terminal domain. Companion studies indicate that an R(98)W mutation in yVps35 causes defective retromer assembly in Saccharomyces cerevisiae. Herein, we find that the expression of hVps35 in yeast confers dominant-negative vacuolar proenzyme secretion and defective secretory proprotein processing. The mutant phenotype appears to be driven by hVps35 competing with endogenous yVps35, becoming incorporated into defective retromer complexes and causing proteasomal degradation of endogenous Vps26 and Vps29. Increased expression of yVps35 displaces some hVps35 to a 100 000 x g supernatant and suppresses the dominant-negative phenotype. Remarkably, mutation of the conserved R(107)W of hVps35 displaces some of the protein to the 100 000 x g supernatant, slows protein turnover and restores stability of Vps26p and Vps29p and completely abrogates dominant-negative trafficking behavior. We show that hVps35 coprecipitates Vps26, whereas the R(107)W mutant does not. In pancreatic beta cells, the R(107)W mutant shifts hVps35 from peripheral endosomes to a juxtanuclear compartment, affecting both mannose phosphate receptors and insulin. These data underscore importance of the Vps35 PRLYL motif in retromer subcomplex interactions and function. PMID- 17916229 TI - Effects of selective COX-2 inhibition on prostanoids and platelet physiology in young healthy volunteers. AB - BACKGROUND: Selective inhibitors of cyclooxygenase-2 (COX-2) called coxibs, are effective anti-inflammatory and analgesic drugs. Recently, these drugs were associated with an increased risk for myocardial infarction and atherothrombotic events. The hypothesis of thromboxane-prostacyclin imbalance has been preferred to explain these unwanted effects. METHODS: We studied the effects of 14 days intake of rofecoxib (25 mg q.d.), celecoxib (200 mg b.i.d.), naproxen (500 mg b.i.d.) and placebo in a randomized, blinded, placebo-controlled study in young healthy volunteers (median age 25-30 years, each group n = 10). We assessed prostanoid metabolite excretion (PGE-M, TXB(2), 6-keto-PGF(1alpha), 11-dehydro TXB(2), 2,3-dinor-TXB(2), and dinor-6-keto-PGF(1alpha)), the expression of platelet activation markers (CD62P, PAC-1, fibrinogen), platelet-leukocyte formation, the endogenous thrombin potential, platelet cAMP content and plasma thrombomodulin level. RESULTS: Naproxen suppressed biosynthesis of PGE-M, prostacyclin metabolites and thromboxane metabolites and thrombomodulin levels. In contrast, both coxibs had an inhibitory effect only on PGE-M, 6-keto PGF(1alpha), and on dinor-6-keto-PGF(1alpha), whereas TXB(2), 2,3-dinor-TXB(2) and 11-dehydro-TXB(2) excretion were unaffected. None of the coxibs exerted significant effects on the expression of platelet activation markers, cAMP generation, platelet-leukocyte formation, or on thrombomodulin plasma levels. Interestingly, platelet TXB(2) release during aggregation was enhanced after coxib treatment following arachidonic acid or collagen stimulation. CONCLUSION: In young healthy volunteers coxibs inhibit systemic PGE(2) and PGI(2) synthesis. Platelet function and expression of platelet aggregation markers are not affected; however, coxibs can stimulate TXB(2) release from activated platelets. Combined decrease in vasodilatory PGE(2) and PGI(2) together with increased TXA(2) in proaggregatory conditions may contribute to coxib side effects. PMID- 17916230 TI - LPS induces IL-10 production by human alveolar macrophages via MAPKinases- and Sp1-dependent mechanisms. AB - BACKGROUND: IL-10 is a cytokine mainly produced by macrophages that plays key roles in tolerance to inhaled antigens and in lung homeostasis. Its regulation in alveolar macrophages (HAM), the resident lung phagocytes, remains however unknown. METHODS: The present study investigated the role of intracellular signalling and transcription factors controlling the production of IL-10 in LPS activated HAM from normal nonsmoking volunteers. RESULTS: LPS (1-1000 pg/ml) induced in vitro IL-10 production by HAM, both at mRNA and protein levels. LPS also activated the phosphorylation of ERK, p38 and JNK MAPkinases (immunoblots) and Sp-1 nuclear activity (EMSA). Selective inhibitors of MAPKinases (respectively PD98059, SB203580 and SP600125) and of Sp-1 signaling (mithramycin) decreased IL-10 expression in HAM. In addition, whilst not affecting IL-10 mRNA degradation, the three MAPKinase inhibitors completely abolished Sp-1 activation by LPS in HAM. CONCLUSION: These results demonstrate for the first time that expression of IL-10 in lung macrophages stimulated by LPS depends on the concomitant activation of ERK, p38 and JNK MAPKinases, which control downstream signalling to Sp-1 transcription factor. This study further points to Sp-1 as a key signalling pathway for IL-10 expression in the lung. PMID- 17916232 TI - PAZAR: a framework for collection and dissemination of cis-regulatory sequence annotation. AB - PAZAR is an open-access and open-source database of transcription factor and regulatory sequence annotation with associated web interface and programming tools for data submission and extraction. Curated boutique data collections can be maintained and disseminated through the unified schema of the mall-like PAZAR repository. The Pleiades Promoter Project collection of brain-linked regulatory sequences is introduced to demonstrate the depth of annotation possible within PAZAR. PAZAR, located at http://www.pazar.info, is open for business. PMID- 17916231 TI - A genetic code alteration generates a proteome of high diversity in the human pathogen Candida albicans. AB - BACKGROUND: Genetic code alterations have been reported in mitochondrial, prokaryotic, and eukaryotic cytoplasmic translation systems, but their evolution and how organisms cope and survive such dramatic genetic events are not understood. RESULTS: Here we used an unusual decoding of leucine CUG codons as serine in the main human fungal pathogen Candida albicans to elucidate the global impact of genetic code alterations on the proteome. We show that C. albicans decodes CUG codons ambiguously and tolerates partial reversion of their identity from serine back to leucine on a genome-wide scale. CONCLUSION: Such codon ambiguity expands the proteome of this human pathogen exponentially and is used to generate important phenotypic diversity. This study highlights novel features of C. albicans biology and unanticipated roles for codon ambiguity in the evolution of the genetic code. PMID- 17916233 TI - Changes in BMI-distribution from 1966-69 to 1995-97 in adolescents. The Young HUNT study, Norway. AB - BACKGROUND: The aim of this study was to explore changes in the BMI-distribution over time among Norwegian adolescents. METHODS: Height and weight were measured in standardised ways and BMI computed in 6774 adolescents 14-18 years who participated in the Young-HUNT study, the youth part of the Health-study of Nord Trondelag County, Norway in 1995-97. The results were compared to data from 8378 adolescents, in the same age group and living in the same geographical region, collected by the National Health Screening Service in 1966-69. RESULTS: From 1966 69 to 1995-97 there was an increased dispersion and a two-sided change in the BMI distribution. Mean BMI did not increase in girls aged 14-17, but increased significantly in 18 year old girls and in boys of all ages. In both sexes and all ages there was a significant increase in the upper percentiles, but also a trend towards a decrease in the lowest percentiles. Height and weight increased significantly in both sexes and all ages. CONCLUSION: The increased dispersion of the BMI-distribution with a substantial increase in upper BMI-percentiles followed the same pattern seen in other European countries and the United States. The lack of increase in mean BMI among girls, and the decrease in the lowest percentiles has not been acknowledged in previous studies, and may call for attention. PMID- 17916234 TI - Family expansion and gene rearrangements contributed to the functional specialization of PRDM genes in vertebrates. AB - BACKGROUND: Progressive diversification of paralogs after gene expansion is essential to increase their functional specialization. However, mode and tempo of this divergence remain mostly unclear. Here we report the comparative analysis of PRDM genes, a family of putative transcriptional regulators involved in human tumorigenesis. RESULTS: Our analysis assessed that the PRDM genes originated in metazoans, expanded in vertebrates and further duplicated in primates. We experimentally showed that fast-evolving paralogs are poorly expressed, and that the most recent duplicates, such as primate-specific PRDM7, acquire tissue specificity. PRDM7 underwent major structural rearrangements that decreased the number of encoded Zn-Fingers and modified gene splicing. Through internal duplication and activation of a non-canonical splice site (GC-AG), PRDM7 can acquire a novel intron. We also detected an alternative isoform that can retain the intron in the mature transcript and that is predominantly expressed in human melanocytes. CONCLUSION: Our findings show that (a) molecular evolution of paralogs correlates with their expression pattern; (b) gene diversification is obtained through massive genomic rearrangements; and (c) splicing modification contributes to the functional specialization of novel genes. PMID- 17916235 TI - Radiological informed consent in cardiovascular imaging: towards the medico-legal perfect storm? AB - Use of radiation for medical examinations and tests is the largest manmade source of radiation exposure. No one can doubt the immense clinical and scientific benefits of imaging to the modern practice of medicine. Every radiological and nuclear medicine examination confers a definite (albeit low) long-term risk of cancer, but patients undergoing such examinations often receive no or inaccurate information about radiological dose exposure and corresponding risk directly related to the radiological dose received. Too detailed information on radiological dose and risk may result in undue anxiety, but information "economical with the truth" may violate basic patients' rights well embedded in ethics (Oviedo convention 1997) and law (97/43 Euratom Directive 1997). Informed consent is a procedure needed to establish a respectful and ethical relation between doctors and patients. Nevertheless, in an "ideal" consent process, the principle of patient autonomy in current radiological practice might be reinforced by making it mandatory to obtain explicit and transparent informed consent form for radiological examination with high exposure (>or= 500 chest x rays). The form may spell-out the type of examination, the exposure in effective dose (mSv), derived from reference values in guidelines or - better - from actual values from their department. The dose equivalent might be also expressed in number of chest radiographs and the risk of cancer as number of extra cases in the exposed population, derived from most recent and authorative guidelines (e.g., BEIR VII Committee, release 2006). Common sense, deontological code, patients'rights, medical imaging guidelines, Euratom law, all coherently and concordantly encourage and recommend a justified, optimized, responsible and informed use of testing with ionizing radiation. Although the idea of informed consent for radiation dose does not seem to be on the immediate radar screen at least in the US, the current practice clashes against these guidelines and laws. PMID- 17916236 TI - Hypophosphatasia. AB - Hypophosphatasia is a rare inherited disorder characterized by defective bone and teeth mineralization, and deficiency of serum and bone alkaline phosphatase activity. The prevalence of severe forms of the disease has been estimated at 1/100 000. The symptoms are highly variable in their clinical expression, which ranges from stillbirth without mineralized bone to early loss of teeth without bone symptoms. Depending on the age at diagnosis, six clinical forms are currently recognized: perinatal (lethal), perinatal benign, infantile, childhood, adult and odontohypophosphatasia. In the lethal perinatal form, the patients show markedly impaired mineralization in utero. In the prenatal benign form these symptoms are spontaneously improved. Clinical symptoms of the infantile form are respiratory complications, premature craniosynostosis, widespread demineralization and rachitic changes in the metaphyses. The childhood form is characterized by skeletal deformities, short stature, and waddling gait, and the adult form by stress fractures, thigh pain, chondrocalcinosis and marked osteoarthropathy. Odontohypophosphatasia is characterized by premature exfoliation of fully rooted primary teeth and/or severe dental caries, often not associated with abnormalities of the skeletal system. The disease is due to mutations in the liver/bone/kidney alkaline phosphatase gene (ALPL; OMIM# 171760) encoding the tissue-nonspecific alkaline phosphatase (TNAP). The diagnosis is based on laboratory assays and DNA sequencing of the ALPL gene. Serum alkaline phosphatase (AP) activity is markedly reduced in hypophosphatasia, while urinary phosphoethanolamine (PEA) is increased. By using sequencing, approximately 95% of mutations are detected in severe (perinatal and infantile) hypophosphatasia. Genetic counseling of the disease is complicated by the variable inheritance pattern (autosomal dominant or autosomal recessive), the existence of the uncommon prenatal benign form, and by incomplete penetrance of the trait. Prenatal assessment of severe hypophosphatasia by mutation analysis of chorionic villus DNA is possible. There is no curative treatment for hypophosphatasia, but symptomatic treatments such as non-steroidal anti-inflammatory drugs or teriparatide have been shown to be of benefit. Enzyme replacement therapy will be certainly the most promising challenge of the next few years. PMID- 17916238 TI - Do social networks affect the use of residential aged care among older Australians? AB - BACKGROUND: Older people's social networks with family and friends can affect residential aged care use. It remains unclear if there are differences in the effects of specific (with children, other relatives, friends and confidants) and total social networks upon use of low-level residential care and nursing homes. METHODS: Data were drawn from the Australian Longitudinal Study of Ageing. Six waves of data from 1477 people aged > or = 70 collected over nine years of follow up were used. Multinomial logistic regressions of the effects of specific and total social networks on residential care use were carried out. Propensity scores were used in the analyses to adjust for differences in participant's health, demographic and lifestyle characteristics with respect to social networks. RESULTS: Higher scores for confidant networks were protective against nursing home use (odds ratio [OR] upper versus lower tertile of confidant networks = 0.50; 95%CI 0.33-0.75). Similarly, a significant effect of upper versus lower total network tertile on nursing home use was observed (OR = 0.62; 95%CI 0.43 0.90). Evidence of an effect of children networks on nursing home use was equivocal. Nursing home use was not predicted by other relatives or friends social networks. Use of lower-level residential care was unrelated to social networks of any type. Social networks of any type did not have a significant effect upon low-level residential care use. DISCUSSION: Better confidant and total social networks predict nursing home use in a large cohort of older Australians. Policy needs to reflect the importance of these particular relationships in considering where older people want to live in the later years of life. PMID- 17916237 TI - Functional and evolutionary analysis of alternatively spliced genes is consistent with an early eukaryotic origin of alternative splicing. AB - BACKGROUND: Alternative splicing has been reported in various eukaryotic groups including plants, apicomplexans, diatoms, amoebae, animals and fungi. However, whether widespread alternative splicing has evolved independently in the different eukaryotic groups or was inherited from their last common ancestor, and may therefore predate multicellularity, is still unknown. To better understand the origin and evolution of alternative splicing and its usage in diverse organisms, we studied alternative splicing in 12 eukaryotic species, comparing rates of alternative splicing across genes of different functional classes, cellular locations, intron/exon structures and evolutionary origins. RESULTS: For each species, we find that genes from most functional categories are alternatively spliced. Ancient genes (shared between animals, fungi and plants) show high levels of alternative splicing. Genes with products expressed in the nucleus or plasma membrane are generally more alternatively spliced while those expressed in extracellular location show less alternative splicing. We find a clear correspondence between incidence of alternative splicing and intron number per gene both within and between genomes. In general, we find several similarities in patterns of alternative splicing across these diverse eukaryotes. CONCLUSION: Along with previous studies indicating intron-rich genes with weak intron boundary consensus and complex spliceosomes in ancestral organisms, our results suggest that at least a simple form of alternative splicing may already have been present in the unicellular ancestor of plants, fungi and animals. A role for alternative splicing in the evolution of multicellularity then would largely have arisen by co-opting the preexisting process. PMID- 17916239 TI - All duplicates are not equal: the difference between small-scale and genome duplication. AB - BACKGROUND: Genes in populations are in constant flux, being gained through duplication and occasionally retained or, more frequently, lost from the genome. In this study we compare pairs of identifiable gene duplicates generated by small scale (predominantly single-gene) duplications with those created by a large scale gene duplication event (whole-genome duplication) in the yeast Saccharomyces cerevisiae. RESULTS: We find a number of quantifiable differences between these data sets. Whole-genome duplicates tend to exhibit less profound phenotypic effects when deleted, are functionally less divergent, and are associated with a different set of functions than their small-scale duplicate counterparts. At first sight, either of these latter two features could provide a plausible mechanism by which the difference in dispensability might arise. However, we uncover no evidence suggesting that this is the case. We find that the difference in dispensability observed between the two duplicate types is limited to gene products found within protein complexes, and probably results from differences in the relative strength of the evolutionary pressures present following each type of duplication event. CONCLUSION: Genes, and the proteins they specify, originating from small-scale and whole-genome duplication events differ in quantifiable ways. We infer that this is not due to their association with different functional categories; rather, it is a direct result of biases in gene retention. PMID- 17916240 TI - Curcumin enhances the apoptosis-inducing potential of TRAIL in prostate cancer cells: molecular mechanisms of apoptosis, migration and angiogenesis. AB - BACKGROUND: We have recently shown that curcumin (a diferuloylmethane) inhibits growth and induces apoptosis, and also demonstrated that TRAIL induces apoptosis by binding to specific cell surface death receptors in prostate cancer cells. The objectives of this paper were to investigate the molecular mechanisms by which curcumin enhanced the apoptosis-inducing potential of TRAIL in prostate cancer cells. RESULTS: Curcumin enhanced the apoptosis-inducing potential of TRAIL in androgen-unresponsive PC-3 cells and sensitized androgen-responsive TRAIL resistant LNCaP cells. Curcumin inhibited the expressions of Bcl-2, Bcl-XL, survivin and XIAP, and induced the expressions Bax, Bak, PUMA, Bim, and Noxa and death receptors (TRAIL-R1/DR4 and TRAIL-R2/DR5) in both cell lines. Overexpression of dominant negative FADD inhibited the interactive effects of curcumin and TRAIL on apoptosis. Treatment of these cells with curcumin resulted in activation of caspase-3, and caspase-9, and drop in mitochondrial membrane potential, and these events were further enhanced when combined with TRAIL. Curcumin inhibited capillary tube formation and migration of HUVEC cells and these effects were further enhanced in the presence of MEK1/2 inhibitor PD98059. CONCLUSION: The ability of curcumin to inhibit capillary tube formation and cell migration, and enhance the therapeutic potential of TRAIL suggests that curcumin alone or in combination with TRAIL can be used for prostate cancer prevention and/or therapy. PMID- 17916241 TI - Genetic diversity of the obligate intracellular bacterium Chlamydophila pneumoniae by genome-wide analysis of single nucleotide polymorphisms: evidence for highly clonal population structure. AB - BACKGROUND: Chlamydophila pneumoniae is an obligate intracellular bacterium that replicates in a biphasic life cycle within eukaryotic host cells. Four published genomes revealed an identity of > 99 %. This remarkable finding raised questions about the existence of distinguishable genotypes in correlation with geographical and anatomical origin. RESULTS: We studied the genetic diversity of C. pneumoniae by analysing synonymous single nucleotide polymorphisms (sSNPs) that are under reduced selection pressure. We conducted an in silico analysis of the four sequenced genomes, chose 232 representative sSNPs and analysed the loci in 38 C. pneumoniae isolates. We identified 15 different genotypes that were separated in four major clusters. Clusters were not associated with anatomical or geographical origin. However, animal lineages are basal on the C. pneumomiae phylogeny, suggesting a recent transmission to humans through successive bottlenecks some 150,000 years ago. A lack of detectable variation in 17 isolates emphasizes the extraordinary genetic conservation of this species and the high clonality of the population. Moreover, the largest cluster, which encompasses 80% of all analysed strains, is an extremely young clade, that went through an important population expansion some 3,300 years ago. CONCLUSION: sSNPs have proven useful as a sensitive marker to gain new insights into genetic diversity, population structure and evolutionary history of C. pneumoniae. PMID- 17916242 TI - The BRCA1 Ashkenazi founder mutations occur on common haplotypes and are not highly correlated with anonymous single nucleotide polymorphisms likely to be used in genome-wide case-control association studies. AB - BACKGROUND: We studied linkage disequilibrium (LD) patterns at the BRCA1 locus, a susceptibility gene for breast and ovarian cancer, using a dense set of 114 single nucleotide polymorphisms in 5 population groups. We focused on Ashkenazi Jews in whom there are known founder mutations, to address the question of whether we would have been able to identify the 185delAG mutation in a case control association study (should one have been done) using anonymous genetic markers. This mutation is present in approximately 1% of the general Ashkenazi population and 4% of Ashkenazi breast cancer cases. We evaluated LD using pairwise and haplotype-based methods, and assessed correlation of SNPs with the founder mutations using Pearson's correlation coefficient. RESULTS: BRCA1 is characterized by very high linkage disequilibrium in all populations spanning several hundred kilobases. Overall, haplotype blocks and pair-wise LD bins were highly correlated, with lower LD in African versus non-African populations. The 185delAG and 5382insC founder mutations occur on the two most common haplotypes among Ashkenazim. Because these mutations are rare, even though they are in strong LD with many other SNPs in the region as measured by D-prime, there were no strong associations when assessed by Pearson's correlation coefficient, r (maximum of 0.04 for the 185delAG). CONCLUSION: Since the required sample size is related to the inverse of r, this suggests that it would have been difficult to map BRCA1 in an Ashkenazi case-unrelated control association study using anonymous markers that were linked to the founder mutations. PMID- 17916243 TI - Acute cholecystitis - early laparoskopic surgery versus antibiotic therapy and delayed elective cholecystectomy: ACDC-study. AB - BACKGROUND: Acute cholecystitis occurs frequently in the elderly and in patients with gall stones. Most cases of severe or recurrent cholecystitis eventually require surgery, usually laparoscopic cholecystectomy in the Western World. It is unclear whether an initial, conservative approach with antibiotic and symptomatic therapy followed by delayed elective surgery would result in better morbidity and outcome than immediate surgery. At present, treatment is generally determined by whether the patient first sees a surgeon or a gastroenterologist. We wish to investigate whether both approaches are equivalent. The primary endpoint is the morbidity until day 75 after inclusion into the study. DESIGN: A multicenter, prospective, randomized non-blinded study to compare treatment outcome, complications and 75-day morbidity in patients with acute cholecystitis randomized to laparoscopic cholecystectomy within 24 hours of symptom onset or antibiotic treatment with moxifloxacin and subsequent elective cholecystectomy. For consistency in both arms moxifloxacin, a fluorquinolone with broad spectrum of activity and high bile concentration is used as antibiotic. DURATION: October 2006 - November 2008. ORGANISATION/RESPONSIBILITY: The trial was planned and is being conducted and analysed by the Departments of Gastroenterology and General Surgery at the University Hospital of Heidelberg according to the ethical, regulatory and scientific principles governing clinical research as set out in the Declaration of Helsinki (1989) and the Good Clinical Practice guideline (GCP). TRIAL REGISTRATION: ClinicalTrials.gov NCT00447304. PMID- 17916245 TI - Effects of affective picture viewing on postural control. AB - BACKGROUND: Emotion theory holds that unpleasant events prime withdrawal actions, whereas pleasant events prime approach actions. Recent studies have suggested that passive viewing of emotion eliciting images results in postural adjustments, which become manifest as changes in body center of pressure (COP) trajectories. From those studies it appears that posture is modulated most when viewing pictures with negative valence. The present experiment was conducted to test the hypothesis that pictures with negative valence have a greater impact on postural control than neutral or positive ones. Thirty-four healthy subjects passively viewed a series of emotion eliciting images, while standing either in a bipedal or unipedal stance on a force plate. The images were adopted from the International Affective Picture System (IAPS). We analysed mean and variability of the COP and the length of the associated sway path as a function of emotion. RESULTS: The mean position of the COP was unaffected by emotion, but unipedal stance resulted in overall greater body sway than bipedal stance. We found a modest effect of emotion on COP: viewing pictures of mutilation resulted in a smaller sway path, but only in unipedal stance. We obtained valence and arousal ratings of the images with an independent sample of viewers. These subjects rated the unpleasant images as significantly less pleasant than neutral images, and the pleasant images as significantly more pleasant than neutral images. However, the subjects rated the images as overall less pleasant and less arousing than viewers in a closely comparable American study, pointing to unknown differences in viewer characteristics. CONCLUSION: Overall, viewing emotion eliciting images had little effect on body sway. Our finding of a reduction in sway path length when viewing pictures of mutilation was indicative of a freezing strategy, i.e. fear bradycardia. The results are consistent with current knowledge about the neuroanatomical organization of the emotion system and the neural control of behavior. PMID- 17916244 TI - Genes2Networks: connecting lists of gene symbols using mammalian protein interactions databases. AB - BACKGROUND: In recent years, mammalian protein-protein interaction network databases have been developed. The interactions in these databases are either extracted manually from low-throughput experimental biomedical research literature, extracted automatically from literature using techniques such as natural language processing (NLP), generated experimentally using high-throughput methods such as yeast-2-hybrid screens, or interactions are predicted using an assortment of computational approaches. Genes or proteins identified as significantly changing in proteomic experiments, or identified as susceptibility disease genes in genomic studies, can be placed in the context of protein interaction networks in order to assign these genes and proteins to pathways and protein complexes. RESULTS: Genes2Networks is a software system that integrates the content of ten mammalian interaction network datasets. Filtering techniques to prune low-confidence interactions were implemented. Genes2Networks is delivered as a web-based service using AJAX. The system can be used to extract relevant subnetworks created from "seed" lists of human Entrez gene symbols. The output includes a dynamic linkable three color web-based network map, with a statistical analysis report that identifies significant intermediate nodes used to connect the seed list. CONCLUSION: Genes2Networks is powerful web-based software that can help experimental biologists to interpret lists of genes and proteins such as those commonly produced through genomic and proteomic experiments, as well as lists of genes and proteins associated with disease processes. This system can be used to find relationships between genes and proteins from seed lists, and predict additional genes or proteins that may play key roles in common pathways or protein complexes. PMID- 17916246 TI - Longer pregnancy and slower fetal development in women with latent "asymptomatic" toxoplasmosis. AB - BACKGROUND: The purpose of this study was to confirm that women with latent toxoplasmosis have developmentally younger fetuses at estimated pregnancy week 16 and to test four exclusive hypotheses that could explain the observed data. METHODS: In the present retrospective cohort study we analysed by the GLM (general linear model) method data from 730 Toxoplasma-free and 185 Toxoplasma infected pregnant women. RESULTS: At pregnancy week 16 estimated from the date of the last menstruation, the mothers with latent toxoplasmosis had developmentally younger fetuses based on ultrasound scan (P = 0.014). Pregnancy of Toxoplasma positive compared to Toxoplasma-negative women was by about 1.3 days longer, as estimated both from the date of the last menstruation (P = 0.015) and by ultrasonography (P = 0.025). CONCLUSION: The most parsimonious explanation for the observed data is retarded fetal growth during the first weeks of pregnancy in Toxoplasma-positive women. The phenomenon was only detectable in multiparous women, suggesting that the immune system may play some role in it. PMID- 17916247 TI - Mitochondrial DNA mutations in oxyphilic and chief cell parathyroid adenomas. AB - BACKGROUND: The potential pathogenetic significance of mitochondrial DNA (mtDNA) mutations in tumorigenesis is controversial. We hypothesized that benign tumorigenesis of a slowly replicating tissue like the human parathyroid might constitute an especially fertile ground on which a selective advantage conferred by mtDNA mutation could be manifested and might contribute to the oxyphilic phenotype observed in a subset of parathyroid tumors. METHODS: We sought acquired mitochondrial DNA mutations by sequencing the entire 16.6 kb mitochondrial genome of each of thirty sporadic parathyroid adenomas (18 chief cell and 12 oxyphil cell), eight independent, polyclonal, parathyroid primary chief cell hyperplasias plus corresponding normal control samples, five normal parathyroid glands, and one normal thyroid gland. RESULTS: Twenty-seven somatic mutations were identified in 15 of 30 (9 of 12 oxyphil adenomas, 6 of 18 chief cell) parathyroid adenomas studied. No somatic mutations were observed in the hyperplastic parathyroid glands. CONCLUSION: Features of the somatic mutations suggest that they may confer a selective advantage and contribute to the molecular pathogenesis of parathyroid adenomas. Importantly, the statistically significant differences in mutation prevalence in oxyphil vs. chief cell adenomas also suggest that mtDNA mutations may contribute to the oxyphil phenotype. PMID- 17916250 TI - The Framingham Heart Study, on its way to becoming the gold standard for Cardiovascular Genetic Epidemiology? AB - The Framingham Heart Study, founded in 1948 to examine the epidemiology of cardiovascular disease in a small town outside of Boston, has become the worldwide standard for cardiovascular epidemiology. It is among the longest running, most comprehensively characterized multi-generational studies in the world. Such seminal findings as the effects of smoking and high cholesterol on heart disease came from the Framingham Heart Study. At the time of publication these were novel cardiovascular disease (CVD) risk factors, now they are the basis of treatment and prevention in the US. Is the Framingham study now on it's way to becoming the gold standard for genetic epidemiology of CVD? Will the novel genetic findings of today become the health care standards of tomorrow? The accompanying articles summarizing the results of genome-wide association studies (GWAS) give the reader a first glimpse into the possibilities. PMID- 17916249 TI - Elastic intramedullary nailing and DBM-bone marrow injection for the treatment of simple bone cysts. AB - BACKGROUND: Simple or unicameral bone cysts are common benign fluid-filled lesions usually located at the long bones of children before skeletal maturity. METHODS: We performed demineralized bone matrix and iliac crest bone marrow injection combined with elastic intramedullary nailing for the treatment of simple bone cysts in long bones of 9 children with a mean age of 12.6 years (range, 4 to 15 years). RESULTS: Two of the 9 patients presented with a pathological fracture. Three patients had been referred after the failure of previous treatments. Four patients had large lesions with impending pathological fractures that interfered with daily living activities. We employed a ratio to ascertain the severity of the lesion. The extent of the lesion on the longitudinal axis was divided with the normal expected diameter of the long bone at the site of the lesion. The mean follow-up was 77 months (range, 5 to 8 years). All patients were pain free and had full range of motion of the adjacent joints at 6 weeks postoperatively. Review radiographs showed that all 7 cysts had consolidated completely (Neer stage I) and 2 cysts had consolidated partially (Neer stage II). Until the latest examination there was no evidence of fracture or re-fracture. CONCLUSION: Elastic intramedullary nailing has the twofold benefits of continuous cyst decompression, and early immediate stability to the involved bone segment, which permits early mobilization and return to the normal activities of the pre-teen patients. PMID- 17916248 TI - Wide diversity in structure and expression profiles among members of the Caenorhabditis elegans globin protein family. AB - BACKGROUND: The emergence of high throughput genome sequencing facilities and powerful high performance bioinformatic tools has highlighted hitherto unexpected wide occurrence of globins in the three kingdoms of life. In silico analysis of the genome of C. elegans identified 33 putative globin genes. It remains a mystery why this tiny animal might need so many globins. As an inroad to understanding this complexity we initiated a structural and functional analysis of the globin family in C. elegans. RESULTS: All 33 C. elegans putative globin genes are transcribed. The translated sequences have the essential signatures of single domain bona fide globins, or they contain a distinct globin domain that is part of a larger protein. All globin domains can be aligned so as to fit the globin fold, but internal interhelical and N- and C-terminal extensions and a variety of amino acid substitutions generate much structural diversity among the globins of C. elegans. Likewise, the encoding genes lack a conserved pattern of intron insertion positioning. We analyze the expression profiles of the globins during the progression of the life cycle, and we find that distinct subsets of globins are induced, or repressed, in wild-type dauers and in daf 2(e1370)/insulin-receptor mutant adults, although these animals share several physiological features including resistance to elevated temperature, oxidative stress and hypoxic death. Several globin genes are upregulated following oxygen deprivation and we find that HIF-1 and DAF-2 each are required for this response. Our data indicate that the DAF-2 regulated transcription factor DAF-16/FOXO positively modulates hif-1 transcription under anoxia but opposes expression of the HIF-1 responsive globin genes itself. In contrast, the canonical globin of C. elegans, ZK637.13, is not responsive to anoxia. Reduced DAF-2 signaling leads to enhanced transcription of this globin and DAF-16 is required for this effect. CONCLUSION: We found that all 33 putative globins are expressed, albeit at low or very low levels, perhaps indicating cell-specific expression. They show wide diversity in gene structure and amino acid sequence, suggesting a long evolutionary history. Ten globins are responsive to oxygen deprivation in an interacting HIF-1 and DAF-16 dependent manner. Globin ZK637.13 is not responsive to oxygen deprivation and regulated by the Ins/IGF pathway only suggesting that this globin may contribute to the life maintenance program. PMID- 17916251 TI - Impact of delayed initiation of erythropoietin in critically ill patients. AB - BACKGROUND: The purpose of this study was to evaluate the impact of recombinant human erythropoietin (rHuEPO) use for anemia of critical illness at a practice site where delayed initiation is common. METHODS: Retrospective medical record review involving patients treated with rHuEPO for anemia of critical illness. Those patients given rHuEPO or diagnosed with end-stage renal disease (ESRD) prior to ICU admission were excluded. The primary endpoints were rHuEPO use and RBC transfusion patterns. RESULTS: Complete data were collected for consecutive admissions of 126 patients. Average age (SD) and APACHE II score were 56.5 (18.6) years and 25 (7.8), respectively. The median ICU (IQR) and hospital length of stay (LOS) were 24 (11.25, 39) and 29 (17, 44.75) days, respectively. Treatment with rHuEPO was started an average of 12.5 +/- 10.5 days after ICU admission and given for 3.8 +/- 3.8 doses. Eighty percent of patients were transfused with an average total of 5.42 +/- 5.08 units received. RBC exposure inversely correlated with a lower mean hemoglobin response to rHuEPO. ICU LOS (p < 0.0001), hemoglobin at 24 hours (p = 0.055), transfusion within 48 hours of admit (p < 0.0001), and postoperative status (p = 0.019) were the best predictors of transfusion requirements (r2 = 0.37). CONCLUSION: Delayed initiation of rHuEPO for anemia of critical illness resulted in comparable hemoglobin and transfusion benefits. Future studies are needed to establish clinical benefit and role in therapy. RBC exposure may blunt the erythropoietic effects of rHuEPO, potentially frustrating benefits to those of greatest apparent need. PMID- 17916252 TI - Perturbation of gene expression of the chromatin remodeling pathway in premature newborns at risk for bronchopulmonary dysplasia. AB - BACKGROUND: One-third to one-half of all infants born before the 28th week of gestation develop bronchopulmonary dysplasia (BPD). Inflammatory regulators appear to be involved in the pathogenesis of BPD, possibly beginning in fetal life. To evaluate the feasibility of using expression profiling in umbilical cord tissue to discover molecular signatures for developmental staging and for determining risk of BPD, we conducted a cross-sectional study of infants born at less than 28 weeks of gestation (n = 54). Sections of umbilical cord were obtained at birth from 20 infants who later developed BPD and from 34 of their peers who did not develop BPD. RESULTS: Umbilical cord expression profiles at birth exhibited systematic differences in bioenergetic pathways with respect to gestational age. Infants who subsequently developed BPD had distinct signatures involving chromatin remodeling and histone acetylation pathways, which have previously been implicated in several adult onset lung diseases. These findings are consistent with prior work on inflammatory processes and bioenergetics in prematurity. CONCLUSION: This study of gene expression of the newborn umbilical cord implicates the chromatin remodeling pathways in those premature infants who subsequently develop BPD. Larger sample sizes will be required to generate prognostic markers from umbilical cord profiles. PMID- 17916253 TI - Lack of cortisol response in patients with posttraumatic stress disorder (PTSD) undergoing a diagnostic interview. AB - BACKGROUND: According to DSM-IV, the diagnosis of posttraumatic stress disorder (PTSD) requires the experience of a traumatic event during which the person's response involved intense fear, helplessness, or horror. In order to diagnose PTSD, clinicians must interview the person in depth about his/her previous experiences and determine whether the individual has been traumatized by a specific event or events. However, asking questions about traumatic experiences can be stressful for the traumatized individual and it has been cautioned that subsequent "re-traumatization" could occur. This study investigated the cortisol response in traumatized refugees with PTSD during a detailed and standardized interview about their personal war and torture experiences. METHODS: Participants were male refugees with severe PTSD who solicited an expert opinion in the Psychological Research Clinic for Refugees of the University of Konstanz. 17 patients were administered the Vivo Checklist of War, Detention, and Torture Events, a standardized interview about traumatic experiences, and 16 subjects were interviewed about absorption behavior. Self-reported measures of affect and arousal, as well as saliva cortisol were collected at four points. Before and after the experimental intervention, subjects performed a Delayed Matching-to Sample (DMS) task for distraction. They also rated the severity of selected PTSD symptoms, as well as the level of intrusiveness of traumatic memories at that time. RESULTS: Cortisol excretion diminished in the course of the interview and showed the same pattern for both groups. No specific response was detectable after the supposed stressor. Correspondingly, ratings of subjective well-being, memories of the most traumatic event(s) and PTSD symptoms did not show any significant difference between groups. Those in the presumed stress condition did not perform worse than persons in the control condition after the stressor. However, both groups performed poorly in the DMS task, which is consistent with memory and concentration problems demonstrated in patients with PTSD. CONCLUSION: A comprehensive diagnostic interview including questions about traumatic events does not trigger an HPA-axis based alarm response or changes in psychological measures, even for persons with severe PTSD, such as survivors of torture. Thus, addressing traumatic experiences within a safe and empathic environment appears to impose no unacceptable additional load to the patient. PMID- 17916254 TI - Primary care patients in psychiatric clinical trials: a pilot study using videoconferencing. AB - BACKGROUND: While primary care physicians play a pivotal role in the treatment of depression, collaboration between primary care and psychiatry in clinical research has been limited. Primary care settings provide unique opportunities to improve the methodology of psychiatric clinical trials, by providing more generalizable and less treatment-resistant patients. We examined the feasibility of identifying, recruiting, screening and assessing primary care patients for psychiatric clinical trials using high-quality videoconferencing in a mock clinical trial. METHODS: 1329 patients at two primary care clinics completed a self-report questionnaire. Those screening positive for major depression, panic, or generalized anxiety were given a diagnostic interview via videoconference. Those eligible were provided treatment as usual by their primary care physician, and had 6 weekly assessments by the off-site clinician via videoconferencing. RESULTS: 45 patients were enrolled over 22 weeks, with 36 (80%) completing the six-week study with no more than two missed appointments. All diagnostic groups improved significantly; 94% reported they would participate again, 87% would recommend participation to others, 96% felt comfortable communicating via videoconference, and 94% were able to satisfactorily communicate their feelings via video. CONCLUSION: Results showed that primary care patients will enroll, participate in and complete psychiatric research protocols using remote interviews conducted via videoconference. PMID- 17916255 TI - Androgen-induced masculinization in rainbow trout results in a marked dysregulation of early gonadal gene expression profiles. AB - BACKGROUND: Fish gonadal sex differentiation is affected by sex steroids treatments providing an efficient strategy to control the sexual phenotype of fish for aquaculture purposes. However, the biological effects of such treatments are poorly understood. The aim of this study was to identify the main effects of an androgen masculinizing treatment (11beta-hydroxyandrostenedione, 11betaOHDelta4, 10 mg/kg of food for 3 months) on gonadal gene expression profiles of an all-female genetic population of trout. To characterize the most important molecular features of this process, we used a large scale gene expression profiling approach using rainbow trout DNA microarrays combined with a detailed gene ontology (GO) analysis. RESULTS: 2,474 genes were characterized as up-regulated or down-regulated in trout female gonads masculinized by androgen in comparison with control male or female gonads from untreated all-male and all female genetic populations. These genes were classified in 13 k-means clusters of temporally correlated expression profiles. Gene ontology (GO) data mining revealed that androgen treatment triggers a marked down-regulation of genes potentially involved in early oogenesis processes (GO 'mitotic cell cycle', 'nucleolus'), an up-regulation of the translation machinery (GO 'ribosome') along with a down-regulation of proteolysis (GO 'proteolysis', 'peptidase' and 'metallopeptidase activity'). Genes considered as muscle fibres markers (GO 'muscle contraction') and genes annotated as structural constituents of the extracellular matrix (GO 'extracellular matrix') or related to meiosis (GO 'chromosome' and 'meiosis') were found significantly enriched in the two clusters of genes specifically up-regulated in androgen-treated female gonads. GO annotations 'Sex differentiation' and 'steroid biosynthesis' were enriched in a cluster of genes with high expression levels only in control males. Interestingly none of these genes were stimulated by the masculinizing androgen treatment. CONCLUSION: This study provides evidence that androgen masculinization results in a marked dysregulation of early gene expression profiles when compared to natural testicular or ovarian differentiation. Based on these results we suggest that, in our experimental conditions, androgen masculinization proceeds mainly through an early inhibition of female development. PMID- 17916256 TI - Taking stock of current societal, political and academic stakeholders in the Canadian healthcare knowledge translation agenda. AB - BACKGROUND: In the past 15 years, knowledge translation in healthcare has emerged as a multifaceted and complex agenda. Theoretical and polemical discussions, the development of a science to study and measure the effects of translating research evidence into healthcare, and the role of key stakeholders including academe, healthcare decision-makers, the public, and government funding bodies have brought scholarly, organizational, social, and political dimensions to the agenda. OBJECTIVE: This paper discusses the current knowledge translation agenda in Canadian healthcare and how elements in this agenda shape the discovery and translation of health knowledge. DISCUSSION: The current knowledge translation agenda in Canadian healthcare involves the influence of values, priorities, and people; stakes which greatly shape the discovery of research knowledge and how it is or is not instituted in healthcare delivery. As this agenda continues to take shape and direction, ensuring that it is accountable for its influences is essential and should be at the forefront of concern to the Canadian public and healthcare community. This transparency will allow for scrutiny, debate, and improvements in health knowledge discovery and health services delivery. PMID- 17916257 TI - Across-province standardization and comparative analysis of time-to-care intervals for cancer. AB - BACKGROUND: A set of consistent, standardized definitions of intervals and populations on which to report across provinces is needed to inform the Provincial/Territorial Deputy Ministries of Health on progress of the Ten-Year Plan to Strengthen Health Care. The objectives of this project were to: 1) identify a set of criteria and variables needed to create comparable measures of important time-to-cancer-care intervals that could be applied across provinces and 2) use the measures to compare time-to-care across participating provinces for lung and colorectal cancer patients diagnosed in 2004. METHODS: A broad-based group of stakeholders from each of the three participating cancer agencies was assembled to identify criteria for time-to-care intervals to standardize, evaluate possible intervals and their corresponding start and end time points, and finalize the selection of intervals to pursue. Inclusion/exclusion criteria were identified for the patient population and the selected time points to reduce potential selection bias. The provincial 2004 colorectal and lung cancer data were used to illustrate across-province comparisons for the selected time-to-care intervals. RESULTS: Criteria identified as critical for time-to-care intervals and corresponding start and end points were: 1) relevant to patients, 2) relevant to clinical care, 3) unequivocally defined, and 4) currently captured consistently across cancer agencies. Time from diagnosis to first radiation or chemotherapy treatment and the smaller components, time from diagnosis to first consult with an oncologist and time from first consult to first radiation or chemotherapy treatment, were the only intervals that met all four criteria. Timeliness of care for the intervals evaluated was similar between the provinces for lung cancer patients but significant differences were found for colorectal cancer patients. CONCLUSION: We identified criteria important for selecting time to-care intervals and appropriate inclusion criteria that were robust across the agencies that did not result in an overly selective sample of patients to be compared. Comparisons of data across three provinces of the selected time-to-care intervals identified several important differences related to treatment and access that require further attention. Expanding this collaboration across Canada would facilitate improvement of and equitable access to quality cancer care at a national level. PMID- 17916258 TI - The impact of migraine prevention on daily activities: a longitudinal and responder analysis from three topiramate placebo-controlled clinical trials. AB - BACKGROUND: Topiramate is approved for the prophylaxis (prevention) of migraine headache in adults. The most common adverse events in the three pivotal, randomized, double-blind, placebo-controlled trials were paresthesia, fatigue, cognitive impairment, anorexia, nausea, and taste alteration. In these trials, topiramate 100 mg/d significantly improved Migraine-Specific Questionnaire (MSQ) scores versus placebo (p < 0.001). The MSQ measures how much migraine limits/interrupts daily performance. Pooled analyses of pivotal trial data were conducted to further assess how topiramate 100 mg/d affects daily activities and patient functioning. METHODS: Mean MSQ and Medical Outcome Study Short Form 36 (SF-36) change scores (baseline to each double-blind assessment point) were calculated for pooled intent-to-treat (ITT) patients. Additionally, pooled ITT patients receiving topiramate 100 mg/d or placebo were combined and divided into two responder groups according to percent reduction in monthly migraine frequency: < 50% responders or >or= 50% responders. Between-group differences were assessed using analysis of covariance. RESULTS: Of 756 patients (mean age 39.8 years, 86% female), 384 received topiramate 100 mg/d and 372 placebo. Topiramate significantly improved all three MSQ domains throughout the double blind phase versus placebo (p = 0.024 [week 8], p < 0.001 [weeks 16 and 26] for role prevention; p < 0.001 for role restriction and emotional function [all time points]). Topiramate 100 mg/d significantly improved SF-36 physical component scores (PCS) throughout the double-blind phase versus placebo (p < 0.001, all time points) and significantly improved mental component scores (MCS) at week 26 (p = 0.043). The greatest topiramate-associated improvements on SF-36 subscales were seen for bodily pain and general health perceptions (p < 0.05; weeks 8, 16, and 26), and physical functioning, vitality, role-physical, and social functioning (p < 0.05; weeks 16 and 26). Significantly greater improvements in all three MSQ domains, as well as the PCS and MCS of SF-36, were observed for >or= 50% responders versus < 50% responders (p < 0.001). Significantly greater percentages of topiramate-treated patients were >or= 50% responders versus placebo (46% versus 23%; p < 0.001). CONCLUSION: Topiramate 100 mg/d significantly improved daily activities and patient functioning at all time points throughout the double-blind phase. Daily function and health status significantly improved for those achieving a >or= 50% migraine frequency reduction. PMID- 17916259 TI - A randomised controlled trial of the effects of a web-based PSA decision aid, Prosdex. Protocol. AB - BACKGROUND: Informed decision making is the theoretical basis in the UK for men's decisions about Prostate Specific Antigen (PSA) testing for prostate cancer testing. The aim of this study is to evaluate the effect of a web-based PSA decision-aid, Prosdex, on informed decision making in men. The objective is to assess the effect of Prosdex on six specific outcomes: (i) knowledge of PSA and prostate cancer-related issues - the principal outcome of the study; (ii) attitudes to testing; (iii) decision conflict; (iv) anxiety; (v) intention to undergo PSA testing; (vi) uptake of PSA testing. In addition, a mathematical simulation model of the effects of Prosdex will be developed. METHODS: A randomised controlled trial with four groups: two intervention groups, one viewing Prosdex and the other receiving a paper version of the site; two control groups, the second controlling for the potential Hawthorn effect of the questionnaire used with the first control group. Men between the ages of 50 and 75, who have not previously had a PSA test, will be recruited from General Practitioners (GPs) in Wales, UK. The principal outcome, knowledge, and four other outcome measures - attitudes to testing, decision conflict, anxiety and intention to undergo testing - will be measured with an online questionnaire, used by men in three of the study groups. Six months later, PSA test uptake will be ascertained from GP records; the online questionnaire will then be repeated. These outcomes, and particularly PSA test uptake, will be used to develop a mathematical simulation model, specifically to consider the impact on health service resources. PMID- 17916260 TI - Interpretation of uniocular and binocular trials of glaucoma medications: an observational case series. AB - BACKGROUND: To predict the effectiveness of topical glaucoma medications based on initial uniocular and binocular treatment. To test a traditional hypothesis that effectiveness following a uniocular trial is associated with the change in IOP in the initially treated eye minus the change in the initially untreated eye. To determine whether uniocular or binocular treatment trials are superior. METHODS: Based on a review of medical records, we identified 168 instances in 154 patients with bilateral primary open angle glaucoma of initial uniocular use of a topical glaucoma medication with well-documented intraocular pressure (IOP) readings at baseline (IOP(A)), during the trial (IOP(B)), and at follow-up (IOP(C)). Abstracted data included demographic data, IOP, and medication use. Predictors of the IOP following the trial (IOP(C)) in each eye were identified by multivariable linear regression. In 70 cases, the predictive ability of initial uniocular and binocular treatment could be directly compared. RESULTS: In a multivariable analysis, the follow-up pressure in the initially treated eye (IOP(1C)) was directly correlated with treated eye IOP during initial uniocular use (IOP(1B), p < 0.001). In a multivariable analysis, the follow-up pressure in the initially untreated eye (IOP(2C)) was directly correlated with its baseline IOP(2A) (p < 0.001), and also tended to be associated with treated IOP(1B) (p = 0.07). The multivariable regression coefficient (b) for the IOP change in the initially untreated eye was generally not close to the value of -1 expected by the classic teaching (for eye 1, b = 0.04, p = 0.35; for eye 2, b = 0.07, p = 0.50). In 70 cases, the uniocular and binocular trials predicted a similar fraction of the variance in follow-up IOP(1C) (r(2) = 0.56 and 0.57, respectively) and IOP(2C) (r(2) = 0.39 and 0.38, respectively). CONCLUSION: 1) For uniocular trials, the IOP change in the untreated eye should not be subtracted from that in the treated eye. 2) Uniocular and binocular trials have similar predictive value when interpreted correctly. Either may be selected based on clinical circumstances. PMID- 17916261 TI - From transcriptome to biological function: environmental stress in an ectothermic vertebrate, the coral reef fish Pomacentrus moluccensis. AB - BACKGROUND: Our understanding of the importance of transcriptional regulation for biological function is continuously improving. We still know, however, comparatively little about how environmentally induced stress affects gene expression in vertebrates, and the consistency of transcriptional stress responses to different types of environmental stress. In this study, we used a multi-stressor approach to identify components of a common stress response as well as components unique to different types of environmental stress. We exposed individuals of the coral reef fish Pomacentrus moluccensis to hypoxic, hyposmotic, cold and heat shock and measured the responses of approximately 16,000 genes in liver. We also compared winter and summer responses to heat shock to examine the capacity for such responses to vary with acclimation to different ambient temperatures. RESULTS: We identified a series of gene functions that were involved in all stress responses examined here, suggesting some common effects of stress on biological function. These common responses were achieved by the regulation of largely independent sets of genes; the responses of individual genes varied greatly across different stress types. In response to heat exposure over five days, a total of 324 gene loci were differentially expressed. Many heat responsive genes had functions associated with protein turnover, metabolism, and the response to oxidative stress. We were also able to identify groups of co regulated genes, the genes within which shared similar functions. CONCLUSION: This is the first environmental genomic study to measure gene regulation in response to different environmental stressors in a natural population of a warm adapted ectothermic vertebrate. We have shown that different types of environmental stress induce expression changes in genes with similar gene functions, but that the responses of individual genes vary between stress types. The functions of heat-responsive genes suggest that prolonged heat exposure leads to oxidative stress and protein damage, a challenge of the immune system, and the re-allocation of energy sources. This study hence offers insight into the effects of environmental stress on biological function and sheds light on the expected sensitivity of coral reef fishes to elevated temperatures in the future. PMID- 17916263 TI - Two methods of assessing the mortality factors affecting the larvae and pupae of Cameraria ohridella in the leaves of Aesculus hippocastanum in Switzerland and Bulgaria. AB - The horse-chestnut leaf miner, Cameraria ohridella, is an invasive alien species defoliating horse-chestnut, a popular ornamental tree in Europe. This paper presents quantitative data on mortality factors affecting larvae and pupae of the leaf miner in Switzerland and Bulgaria, both in urban and forest environments. Two sampling methods were used and compared: a cohort method, consisting of the surveying of pre-selected mines throughout their development, and a grab sampling method, consisting of single sets of leaves collected and dissected at regular intervals. The total mortality per generation varied between 14 and 99%. Mortality was caused by a variety of factors, including parasitism, host feeding, predation by birds and arthropods, plant defence reaction, leaf senescence, intra specific competition and inter-specific competition with a fungal disease. Significant interactions were found between mortality factors and sampling methods, countries, environments and generation. No mortality factor was dominant throughout the sites, generations and methods tested. Plant defence reactions constituted the main mortality factor for the first two larval stages, whereas predation by birds and arthropods and parasitism were more important in older larvae and pupae. Mortality caused by leaf senescence was often the dominant mortality factor in the last annual generation. The cohort method detected higher mortality rates than the grab sampling method. In particular, mortality by plant defence reaction and leaf senescence were better assessed using the cohort method, which is, therefore, recommended for life table studies on leaf miners. PMID- 17916262 TI - F2 screen for resistance to a Bacillus thuringiensis-maize hybrid in the sugarcane borer (Lepidoptera: Crambidae). AB - A novel F2 screening technique was developed for detecting resistance in sugarcane borer, Diatraea saccharalis (F.), to transgenic Bacillus thuringiensis (Bt)-maize expressing the Cry1Ab insecticidal protein. The F2 screening method involved (i) collecting larvae from maize fields; (ii) establishing two-parent families; (iii) screening F2 neonates for survival on Bt-maize leaf tissues; and (iv) confirming resistance on commercial Bt-maize plants. With the F2 screening method, 213 iso-line families of D. saccharalis were established from field collections in northeast Louisiana, USA and were screened for Bt resistance. One family was confirmed to carry a major Bt resistance allele(s). In a laboratory bioassay, larval mortality of the Bt-resistant D. saccharalis on Bt-maize leaf tissues was significantly lower than that of a Bt-susceptible strain. This Bt resistant D. saccharalis population is the first corn stalk borer species that has completed larval development on commercial Bt-maize. The F2 screening protocol developed in this study could be modified for detecting Bt resistance alleles in other similar corn stalk borers, such as the European corn borer, Ostrinia nubilalis (Hubner), and the southwestern corn borer, D. grandiosella Dyar. PMID- 17916264 TI - Effects of kaolin particle film on Myzus persicae (Hemiptera: Aphididae) behaviour and performance. AB - The emergence of resistance mechanisms to, and revocation of, many insecticides used in the control of the polyphagus aphid pest, Myzus persicae (Sulzer), has increased the pressure to develop novel approaches for the control of the pest in many crops. Kaolin-based particle films provide a physical barrier against insect pests and show considerable potential for controlling M. persicae. We conducted a series of laboratory experiments to investigate the mode of action of kaolin against aphids. The material appeared to have no direct effect on M. persicae; spraying adult aphids with aqueous kaolin suspension had no significant impact on their subsequent survival or reproduction on untreated plants. Similarly, when aphids were placed on kaolin-treated host-plants (Brassica oleracea), their performance (survival, growth rate and reproduction) was not significantly different from aphids on untreated plants. However, when M. persicae were given a choice between kaolin-treated and untreated (or water solvent-treated) leaf areas, both adults and nymphs exhibited a significant preference for non-kaolin treated host-plant material. Rejection of kaolin-treated plant material occurred very rapidly (within 20 min) and this behavioural effect may be related to the efficacy of kaolin in controlling aphids under field conditions. PMID- 17916265 TI - Molecular differences in the mitochondrial cytochrome oxidase I (mtCOI) gene and development of a species-specific marker for onion thrips, Thrips tabaci Lindeman, and melon thrips, T. palmi Karny (Thysanoptera: Thripidae), vectors of tospoviruses (Bunyaviridae). AB - A quick and developmental-stage non-limiting method of the identification of vectors of tospoviruses, such as Thrips tabaci and T. palmi, is important in the study of vector transmission, insecticide resistance, biological control, etc. Morphological identification of these thrips vectors is often a stumbling block in the absence of a specialist and limited by polymorphism, sex, stage of development, etc. Molecular identification, on the other hand, is not hampered by the above factors and can easily be followed by a non-specialist with a little training. The mitochondrial cytochrome oxidase I (mtCOI) exhibits reliable inter species variations as compared to the other markers. In this communication, we present the differences in the mtCOI partial sequence of morphologically identified specimens of T. tabaci and T. palmi collected from onion and watermelon, respectively. Species-specific markers, identified in this study, could successfully determine T. tabaci and T. palmi, which corroborated the morphological identification. Phylogenetic analyses showed that both T. tabaci and T. palmi formed different clades as compared to the other NCBI accessions. The implication of these variations in vector efficiency has to be investigated further. The result of this investigation is useful in the quick identification of T. tabaci and T. palmi, a critical factor in understanding the epidemiology of the tospoviruses, their management and also in quarantine. PMID- 17916266 TI - Copulation behaviour of Glossina pallidipes (Diptera: Muscidae) outside and inside the female, with a discussion of genitalic evolution. AB - If species-specific male genitalia are courtship devices under sexual selection by cryptic female choice, then species-specific aspects of the morphology and behaviour of male genitalia should often function to stimulate the female during copulation. The morphology and behaviour of the complex, species-specific male genitalia of the tsetse fly, Glossina pallidipes Austen, were determined from both direct observations and dissections of flash-frozen copulating pairs; we found that some male genitalic traits probably function to stimulate the female, while others function to restrain her. The male clamps the ventral surface of the female's abdomen tightly with his powerful cerci. Clamping does not always result in intromission. Clamping bends the female's body wall and her internal reproductive tract sharply, posteriorly and dorsally, and pinches them tightly. The male performed sustained, complex, stereotyped, rhythmic squeezing movements with his cerci that were not necessary to mechanically restrain the female and appeared instead to have a stimulatory function. Six different groups of modified setae on and near the male's genitalia rub directly against particular sites on the female during squeezing. The designs of these setae correlate with the force with which they press on the female and the probable sensitivity of the female surfaces that they contact. As expected under the hypothesis that these structures are under sexual selection by female choice, several traits suspected to have stimulatory functions have diverged in G. pallidipes and its close relative, G. longipalpis. Additional male non-genitalic behaviour during copulation, redescribed more precisely than in previous publications, is also likely to have a courtship function. The elaborate copulatory courtship behaviour and male genitalia may provide the stimuli that previous studies showed to induce female ovulation and resistance to remating. PMID- 17916267 TI - How can alien species inventories and interception data help us prevent insect invasions? AB - Information relevant to invasion processes and invasive alien insect species management in Central Europe was extracted from two databases: a compilation of two inventories of alien insects in Austria and Switzerland, and a list of interceptions of non-indigenous plant pests in Europe gathered by the European and Mediterranean Plant Protection Organisation (EPPO) for the period 1995-2004. For one-third of the insects established in Switzerland and Austria, the region of origin is unclear. Others come mainly from North America, Asia and the Mediterranean region. Among the intercepted insects, 40% were associated with commodities from Asia, 32% from Europe and only 2% from North America. Sternorrhyncha, Coleoptera and Psocoptera were particularly well represented in the alien fauna compared to the native fauna. In the interception database, Sternorrhyncha were also well represented but Diptera accounted for the highest number of records. Sap feeders and detritivores were the dominant feeding niches in the alien insect fauna. In contrast, external defoliators, stem borers, gall makers, root feeders, predators and parasitoids were underrepresented. Nearly 40% of the alien insects in Switzerland and Austria live only indoors. Another 15% live outdoors but exclusively or predominantly on exotic plants. Less than 20% are found mainly in 'natural' environments. The majority of introductions of alien insects in Europe are associated with the international trade in ornamental plants. An economic impact was found for 40% of the alien insects in Switzerland and Austria, whereas none is known to have an ecological impact. The implications of these observations for further studies and the management of alien species in Europe are discussed. PMID- 17916269 TI - Differential electroantennogram response of females and males of two parasitoid species to host-related green leaf volatiles and inducible compounds. AB - Parasitoids employ different types of host-related volatile signals for foraging and host-location. Host-related volatile signals can be plant-based, originate from the herbivore host or produced from an interaction between herbivores and their plant host. In order to investigate potential sex- and species-related differences in the antennal response of parasitoids to different host-related volatiles, we compared the electroantennogram (EAG) responses of both sexes of the specialist parasitoid, Microplitis croceipes (Cresson), and the generalist, Cotesia marginiventris (Cresson), to varying doses of selected plant-based host related volatiles: two green leaf volatiles (cis-3-hexenol and hexanal) and three inducible compounds (cis-3-hexenyl acetate, linalool, and (E,E)-alpha-farnesene). Mating had no significant effect on EAG response. Females of both species showed significantly greater EAG responses than conspecific males to green leaf volatiles, which are released immediately after initiation of herbivore feeding damage. In contrast, males showed greater responses than conspecific females to inducible compounds released much later after initial damage. Cotesia marginiventris females and males showed greater EAG responses than counterpart M. croceipes to the tested compounds at various doses, suggesting that the generalist parasitoid shows greater antennal sensitivity than the specialist to the tested host-plant volatiles. These results are discussed in relation to the possible roles of green leaf volatiles and inducible compounds in the ecology of female and male parasitoids. PMID- 17916268 TI - Development of silverleaf assay, protein and nucleic acid-based diagnostic techniques for the quick and reliable detection and monitoring of biotype B of the whitefly, Bemisia tabaci (Gennadius). AB - The aim of this study was to develop and optimize silverleaf bioassay, esterase analysis and PCR-based techniques to distinguish quickly and reliably biotype B of the whitefly, Bemisia tabaci (Gennadius), from Indian indigenous biotypes. Zucchini and squash readily develop silverleaf symptoms upon feeding by the B biotype, but they are not readily available in Indian markets. A local pumpkin variety 'Big' was, therefore, used in silverleaf assay, which developed symptoms similar to those on zucchini and squash and can be used reliably to detect B biotype. Analysis of non-specific esterases of B and the indigenous biotypes indicated both quantitative and qualitative differences in esterase patterns. Two high molecular weight bands were unique to B biotype and they occurred in abundance. These esterases were used to develop quick and field-based novel detection methods for differentiating B from the indigenous biotypes. Development of these simple and cost-effective protocols has wider application as they can be potentially used to identify other agricultural pests. Mitochondrial cytochrome oxidase I gene sequences and randomly amplified polymorphic DNA (RAPD) polymorphisms, generated using the primer OpB11, were also found useful for detecting B. tabaci biotypes. A B biotype-specific RAPD band of 800 bp was sequenced, which was used to a develop sequence characterized amplified region (SCAR) marker. The SCAR marker involved the development of B biotype-specific primers that amplified 550 bp PCR products only from B biotype genomic DNA. Silverleaf assay, esterases, RAPDs or a SCAR marker were used in combination to analyse whitefly samples collected from selected locations in India, and it was found that any of these techniques can be used singly or in combination to detect B biotype reliably. The B biotype was found in southern parts of India but not in the north in 2004-06. PMID- 17916270 TI - Exploring plant responses to aphid feeding using a full Arabidopsis microarray reveals a small number of genes with significantly altered expression. AB - The aim of this study was to determine which Arabidopsis thaliana (L.) genes had significantly altered expression following 2-36 h of infestation by the aphid Myzus persicae (Sulzer). Six biological replicates were performed for both control and treatment at each time point, allowing rigorous statistical analysis of any changes. Only two genes showed altered expression after 2 h (one up- and one down-regulated) while two were down-regulated and twenty three were up regulated at 36 h. The transcript annotation allowed classification of the significantly altered genes into a number of classes, including those involved in cell wall modification, carbon metabolism and signalling. Additionally, a number of genes were implicated in oxidative stress and defence against other pathogens. Five genes could not currently be assigned any function. The changes in gene expression are discussed in relation to current models of plant-insect interactions. PMID- 17916271 TI - Early season natural control of the brown planthopper, Nilaparvata lugens: the contribution and interaction of two spider species and a predatory bug. AB - Nilaparvata lugens Stal, the Brown Planthopper (BPH), is a major pest in rice. The lycosid Pardosa pseudoannulata (Bosenberg & Strand) and the linyphiid Atypena formosana (Oi) are the early dominant predators in irrigated rice. Later, predatory bugs, including the mirid Cyrtorhinus lividipennis Reuter, become dominant. In unsprayed rice, BPH numbers normally remain low. While P. pseudoannulata is known to be a key natural enemy of BPH, the contribution to BPH reduction by the smaller A. formosana is less well known. Due to the size difference, A. formosana may also be an intraguild prey of P. pseudoannulata. To investigate predation on BPH by the two spider species alone, together and in combination with C. lividipennis, two cage experiments were conducted, the first with adult spiders, the second with immature/unmated spiders. In the latter, spiders were introduced with a delay to allow a better establishment of C. lividipennis. In both experiments, BPH numbers were lowest in treatments with more than one predator species present. Intermediate BPH reduction was obtained with P. pseudoannulata followed by A. formosana and C. lividipennis. While P. pseudoannulata can prey upon large nymphs and adult BPH, A. formosana favour early instar BPH, leading to a more even control across instars in treatments with both spider species. Pardosa pseudoannulata numbers increased more with A. formosana or A. formosana and C. lividipennis present. In contrast, P. pseudoannulata reduced A. formosana and C. lividipennis numbers. The presence of C. lividipennis apparently lessened intraguild predation on A. formosana and may be an intraguild prey of A. formosana. Competition for prey may have added to the antagonistic interactions found. From a biological control perspective, the presence of both spiders in early rice is an advantage for the biological control of BPH in rice. In early rice, results show that A. formosana is an important predator of BPH and, like C. lividipennis, can complement control by P. pseudoannulata and serve as intraguild prey for the latter species. PMID- 17916272 TI - Fatty acids in component of milk enhance the expression of the cAMP-response element-binding-protein-binding protein (CBP)/p300 gene in developing rats. AB - Fatty acids in milk are thought to play an important role in intestinal maturation and gene expression in the rat small intestine during the suckling weaning period. In the present study, we determined the jejunal mRNA level of the cAMP-response-element-binding-protein-binding protein (CBP)/p300, which is one of the chromatin remodelling factors and regulates histone acetylation, during the postnatal period in rats. The mRNA level of CBP/p300 was high during the suckling and middle of the weaning period (day 5 to 20) and then declined sharply to a low level at the end of the weaning period and after weaning. In situ hybridisation also showed that CBP/p300 mRNA levels in the villus as well as the basal membrane clearly decreased after weaning. Rat pups at age 17 d, weaned to a high-fat diet, showed higher levels of CBP/p300 mRNA than those weaned to a low-fat diet. Oral administration of caprylic acid, oleic acid and linoleic acid, which are major fatty acid components in milk, induced jejunal CBP/p300 gene expression. The present results suggest that fatty acids in components of milk enhance expression of the CBP/p300 genes in the small intestine. PMID- 17916274 TI - Obesity indices in relation to cardiovascular disease risk factors among young adult female students. AB - The objective of the present study was to compare the percentage of body fat (%BF), BMI, and central fat distribution anthropometric measures as indices of obesity and to assess the respective associations with cardiovascular risk factors in young female students. Subjects were 220 healthy Greek female students. Dual-energy X-ray absorptiometry was used to estimate %BF, anthropometric measurements were obtained and blood samples were analysed for CVD risk factors. Results showed that 48.6 % of students had increased adiposity, while a considerable proportion was characterised by central fat distribution irrespective of the anthropometric index used. The proportion of subjects with at least one metabolic risk factor present was 60.4 %. Although %BF was not associated with any of the CVD risk factors, waist circumference, waist:hip ratio and waist:height ratio were all associated with CVD risk factors. Higher levels of these anthropometric variables demonstrated higher prevalence of CVD risk factors. The lack of association between %BF and CVD risk factors could be attributed to the fact that females with undesirable adiposity had a tendency for the gynaecoid type of obesity. In contrast, the present results suggest that central body fat distribution in young women may reflect increased risk due to high visceral and particularly intra-abdominal fat levels. Recent epidemiological data from Greece show a high prevalence of overweight and obesity in young adults. Therefore, assessing the risk for the presence of CVD risk factors is of particular importance. Central obesity anthropometric indices seem to be valuable screening tools for young women. PMID- 17916273 TI - The efficacy of black tea in ameliorating endothelial function is equivalent to that of green tea. AB - Consumption of tea has been shown to improve endothelial function. It is assumed that catechins are the tea components responsible for these beneficial effects. In black tea, catechin concentrations are significantly lower than in green tea. The present study was designed to compare green and black tea with regard to amelioration of endothelial function. Endothelial function in response to both teas was assessed in bovine aortic endothelial cells (BAEC) and rat aortic rings. To elucidate whether these findings are also applicable to humans, flow-mediated dilation (FMD) and nitro-mediated dilation (NMD) were assessed by ultrasound in twenty-one healthy women before and 2 h after consumption of green and black tea (2 h of FMD and NMD), in comparison with water (control). In BAEC, green and black tea significantly increased endothelial NO synthase activity to the same extent. Similarly, both teas induced comparable endothelial-dependent vasodilation in rat aortic rings. In human subjects, ingestion of green and black tea led to significant increases in FMD: from 5.4 (sd 2.3) to 10.2 (sd 3) % (baseline-adjusted difference (BAD) for 2 h of FMD, green tea v. water: 5.0 (95 % CI 3.0, 7.0) %; P < 0.001) and from 5 (sd 2.6) to 9.1 (sd 3.6) % (BAD for 2 h of FMD, black tea v. water: 4.4 (95 % CI 2.3, 6.5) %; P < 0.001), respectively. The increase in FMD was not significantly different between the two tea preparations (BAD for 2 h of FMD, green tea v. black tea: 0.66 (95 % CI - 0.76, 2.09) %; P = 0.36). NMD did not vary between any of the groups. In conclusion, green and black tea are equally effective in improving endothelial function. PMID- 17916275 TI - A comparison of methods to assess changes in dietary patterns from pregnancy to 4 years post-partum obtained using principal components analysis. AB - Few studies have examined the longitudinal nature of dietary patterns obtained using principal components analysis (PCA); the methods used are inconsistent. This paper investigates the methodologies used to assess stability and changes in such patterns. Pregnant women recorded frequency of consumption of various food items as part of regular self-completed questionnaires in the Avon Longitudinal Study of Parents and Children. This was repeated when their children were 4 years of age; 8953 women provided data at both times. Dietary patterns were identified using PCA and component scores were calculated at each time point. Additional 'applied' scores were created at 4 years using the loadings obtained from the PCA on the pregnancy data. Correlations were similar for each component across the time points, though slightly larger using the applied method. The applied scores were considerably lower on average than those obtained from separate PCA at 4 years. Women's scores decreased on 'health conscious' and 'confectionery' components while 'processed' and 'vegetarian' scores both increased over the 4 year period. In contrast, applied scores were systematically lower for all components. When split into quintiles, weighted kappa was slightly higher between pregnancy and applied 4-year scores compared to the separate scores. In this cohort it was felt that the 'applied' method to obtain scores at the second time point was inappropriate, primarily due to the differences in FFQ between the two time points. We recommend that future studies using such 'applied' scores compare them with cross-sectional scores and consider the implications of any differences. PMID- 17916276 TI - Comparative effects of olive oil-based and soyabean oil-based emulsions on infection rate and leucocyte count in critically ill patients receiving parenteral nutrition. AB - Soyabean oil-based emulsions high in linoleic acid used in parenteral nutrition (PN) could interfere with immune function and may increase the risk of septic complications. Olive oil-based emulsions, high in oleic acid, could have fewer immune effects. We compared the effects of a soyabean oil-based emulsion v. an olive oil-based emulsion on infection rate, appearance of new infection episodes, leucocyte count (peak and evolution), acute-phase proteins, and major health outcomes in intensive care unit (ICU) adult patients receiving PN. The study was designed as an observational, retrospective, single-centre, cohort study in a general ICU. Patients in the SOYA cohort (n 16) received a soyabean oil-based emulsion and patients in the OLIVE cohort (n 23), an olive oil-based emulsion. Both cohorts had similar basal characteristics and received a similar energy load. The SOYA cohort received an oleic acid:linoleic acid ratio of 0.43 and the OLIVE cohort 2.99 (P < 0.001). No differences were observed in infection rate and appearance, acute-phase proteins, and major health outcomes. At the end of PN, blood leucocyte count decreased by 3.25 x 109 cells/l in the SOYA cohort and increased by 4.51 x 109 cells/l in the OLIVE cohort from baseline values (P = 0.036). Peak leucocyte count presented a trend for a higher value in the OLIVE cohort v. the SOYA cohort (18.86 v. 15.28 x 109 cells/l; P = 0.078). The use of an olive oil-based emulsion in PN had no effect on infection, acute-phase proteins, major health outcomes, and presented higher leucocyte count at the end of PN and a trend to higher peak leucocyte count when compared with soyabean oil based emulsion in ICU patients. PMID- 17916277 TI - Walnut extract (Juglans regia L.) and its component ellagic acid exhibit anti inflammatory activity in human aorta endothelial cells and osteoblastic activity in the cell line KS483. AB - Epidemiological studies suggest that the incidence of CVD and postmenopausal osteoporosis is low in the Mediterranean area, where herbs and nuts, among others, play an important role in nutrition. In the present study, we sought a role of walnuts (Juglans regia L.) in endothelial and bone-cell function. As the endothelial cell expression of adhesion molecules has been recognised as an early step in inflammation and atherogenesis, we examined the effect of walnut methanolic extract and ellagic acid, one of its major polyphenolic components (as shown by HPLC analysis), on the expression of vascular cell adhesion molecule (VCAM)-1 and intracellular adhesion molecule (ICAM)-1 in human aortic endothelial cells. After incubating the cells with TNF-alpha (1 ng/ml) in the absence and in the presence of walnut extract (10-200 microg/ml) or ellagic acid (10- 7-10- 5 m), the VCAM-1 and ICAM-1 expression was quantified by cell-ELISA. We further evaluated the effect of walnut extract (10-50 microg/ml), in comparison with ellagic acid (10- 9-10- 6m), on nodule formation in the osteoblastic cell line KS483. Walnut extract and ellagic acid decreased significantly the TNF-alpha induced endothelial expression of both VCAM-1 and ICAM-1 (P < 0.01; P < 0.001). Both walnut extract (at 10-25 microg/ml) and ellagic acid (at 10- 9-10- 8 m) induced nodule formation in KS483 osteoblasts. The present results suggest that the walnut extract has a high anti-atherogenic potential and a remarkable osteoblastic activity, an effect mediated, at least in part, by its major component ellagic acid. Such findings implicate the beneficial effect of a walnut enriched diet on cardioprotection and bone loss. PMID- 17916278 TI - Oxidised cholesterol is more hypercholesterolaemic and atherogenic than non oxidised cholesterol in hamsters. AB - The present study was to test the relative hypercholesterolaemic and atherogenic potency of oxidised cholesterol (OxC) and non-oxidised cholesterol in hamsters. An OxC mixture, prepared by heating pure cholesterol (100 g) at 160 degrees C in air for 72 h, contained 78 % cholesterol and 22 % OxC. Fifty Golden Syrian hamsters were randomly divided into five groups of ten animals and fed the control diet, a 0.05 % cholesterol diet (C-0.05), a 0.10 % cholesterol diet (C 0.1), a 0.05 % OxC mixture diet (OxC-0.05) or a 0.10 % OxC mixture diet (OxC 0.1), respectively. The OxC-0.05 and OxC-0.1 groups were more hypercholesterolaemic and had serum total cholesterol 22 and 12 % higher than the corresponding C-0.05 and C-0.1 hamsters (P < 0.05). The OxC-0.1 group demonstrated greater deposition of cholesterol and had a larger area of atherosclerotic plaque in the aorta than the corresponding C-0.1 hamsters (P < 0.05). Similarly, the aorta in the OxC-0.1 group showed greater inhibition on acetylcholine-induced relaxation compared with that in the C-0.1 hamsters. It was concluded that OxC was much more hypercholesterolaemic and atherogenic than cholesterol. PMID- 17916279 TI - The cross-bridge dynamics during ventricular contraction predicted by coupling the cardiac cell model with a circulation model. AB - The force-velocity (F-V) relationship of filament sliding is traditionally used to define the inotropic condition of striated muscles. A simple circulation model combined with the Laplace heart was developed to get a deeper insight into the relationship between the F-V characteristics and the cardiac ventricular inotropy. The circulation model consists of a preload and an afterload compartments. The linear F-V relationship for filament sliding in the NL model (Negroni and Lascano 1996) was replaced by the exponential F-V relation observed by Piazzesi et al. (2002). We also modified the NL model to a hybrid model to benefit from the Ca(2+) cooperativity described by the Robinson model (Robinson et al. 2002). The model was validated by determining the diastolic ventricular pressure-volume relationship of the Laplace heart and the F-V relation of the new hybrid model. The computed parameters of the cardiac cycle agreed well with the physiological data. Computational results showed that the cross-bridge elongation (h in the NL model) temporally undershot the equilibrium h(c) during the ejection period and overshot it during the rapid refilling phase. Thereby the time course of ejection and refilling was retarded. In a simulation where the velocity of the mobile myosin head (dX/dt) was varied, the systolic peak pressure of the ventricle varied from a minimum value at dX/dt = 0 to a saturating value obtained with a constant h(c), providing in silico evidence for a functional impact of the cross-bridge sliding rate on the ventricular inotropy. PMID- 17916280 TI - Forearm and calf tissue oxygenation in term neonates measured with near-infrared spectroscopy. AB - Peripheral tissue oxygenation has been studied with near-infrared spectroscopy (NIRS) on either the forearm or calf with questionable comparability. The aim was to compare forearm and calf tissue oxygenation in healthy term neonates measured with NIRS. Fractional oxygen extraction, tissue oxygenation index, and mixed venous oxygenation were similar in both extremities, whereas oxygen delivery and oxygen consumption of calf tissue were higher. PMID- 17916281 TI - Spatial knowledge of geographical globes: evidence of vertical alignment effects. AB - This study investigated human spatial memory of geographical globes. In two experiments, participants studied locations presented on a three-dimensional globe. Subsequently, participants' knowledge of the locations was tested employing two types of pointing task. Directional judgments from imagined locations on the globe were performed either vertically through the ground (as if digging a straight tunnel between the locations) or horizontally along the surface of the globe (the shortest distance for an aircraft to fly to a given destination). In the vertical pointing task, judgments originating from imagined locations in the upper hemisphere were superior by comparison with those originating from imagined locations in the lower hemisphere. Performance in the horizontal pointing task was more variable, with a tendency for superior judgments originating from imagined locations in the lower hemisphere. The results provide evidence of a novel alignment effect involving vertical judgments. PMID- 17916282 TI - [Colorectal surgery: justification for a specific area of knowledge]. PMID- 17916283 TI - [Body packer: review and experience in a referral hospital]. AB - Smuggling of illicit drugs by concealing them within the human body (body packer) is a medical-legal issue that has increased in the last few decades. Physicians, especially those working in the emergency department, should be familiar with the diagnostic and therapeutic management -usually conservative management- of this type of patient and their possible complications. The present article reviews the general concepts and physiopathology associated with transport of packages in the digestive tract and describes the experience of a referral hospital with a protocol specifically designed for these patients. PMID- 17916284 TI - [Consensus meeting on sentinel lymph node biopsy in breast cancer. Spanish society of mastology and breast disease]. AB - Sentinel lymph node biopsy is a useful diagnostic technique in the management of breast cancer and is widely used and accepted in clinical practice. The results of this technique allow adequate staging with lower associated morbidity. However, at present, there are numerous methodological issues that remain to be resolved in on-going trials and investigations and Consensus Meetings are required to standardize the methodological variations and indications of this procedure. The conclusions of the Consensus Meeting held in Murcia, Spain, organized by The Spanish Society of Mastology and Breast Disease, are reported in this document. PMID- 17916285 TI - [Experience with the Bogota bag technique for temporary abdominal closure]. AB - INTRODUCTION: The Bogota bag technique is a reliable method for open abdominal closure. The aim of this study was to describe our experience with this technique. MATERIAL AND METHOD: We retrospectively evaluated our experience with the Bogota Bag technique between January 2000 and March 2006. Descriptive statistical techniques were applied and percentages and means were calculated. RESULTS: The Bogota bag technique was applied in 12 patients. The technique was the preferred closure system to prevent abdominal compartment syndrome in 11 patients (91.66%) and was required to treat abdominal compartment syndrome in one patient (8.34%). No complications occurred in relation to placement or withdrawal of the Bogota bag. There were no intestinal fistulas or intra-abdominal abscesses. The mean length of hospital stay was 46.33 days and the mean length of stay in the intensive care unit was 16.58 days. The survival rate was 41.66%. CONCLUSIONS: In our experience, the Bogota bag is a useful technique and is the preferred closure system to prevent or treat abdominal compartment syndrome. The high mortality rates described are due to the underlying diseases leading to open abdominal closure and not directly to the Bogota bag technique itself. PMID- 17916286 TI - [Evaluation of parathyroid function in presternal subcutaneous grafting after total parathyroidectomy for renal hyperparathyroidism]. AB - INTRODUCTION: In the setting of total parathyroidectomy (TPT) and parathyroid transplantation (PTx) for renal hyperparathyroidism (RHP), we evaluated long-term parathyroid graft function after subcutaneous presternal transplantation (SCPTx). HYPOTHESIS: Parathyroid glands are surrounded by fatty tissue. Therefore, we postulated that subcutaneous implantation of parathyroid tissue after TPT for RHP could be at least as effective as intramuscular grafting and would avoid the complications of the latter technique. MATERIAL AND METHOD: We performed a study in a university hospital and its dialysis unit. DESIGN: Prospective open efficacy study of a postoperative diagnostic monitoring method of intact parathyroid hormone (iPTH) in a cohort of surgical patients without loss to follow-up. PATIENTS AND INTERVENTIONS: Thirty-five patients (19 women and 16 men) underwent TPT and SCPTx for RHP at the Department of General Surgery and Department of Nephrology, Donostia Hospital, San Sebastian, Gipuzkoa, Spain, from January 2002 to December 2005. Follow-up ranged from 6 to 42 months (mean, 15.4 months). Graft function was evaluated by measurement of plasma iPTH levels before surgery and 24 hours and 1, 3, 5, 15, 30, 60, 100 and 150 weeks after surgery. Reference values for PTH in our laboratory were 20-65 pg/mL. RESULTS: The mean preoperative iPTH values were 1245 +/- 367.9 pg/mL (mean +/- SD) (range, 493-2160). After TPT and SCPTx, iPTH levels became undetectable in all patients at 24 hours. A value of 50 pg/mL was established as the criterion for adequate parathyroid graft function. The following values were obtained: 15.54 +/- 10.61 pg/mL (mean +/- SD) (range, 6 44) after 1 week, 57.2 +/- 1.9 pg/mL (mean +/- SD) (range, 43-74) after 5 weeks, 64.21 +/- 9.73 pg/mL (mean +/- SD) (range, 11.3-89) after 15 weeks, 75.12 +/- 9.05 pg/mL (mean +/- SD) (range, 24.6-104.2) after 30 weeks, 101.63 +/- 19.85 pg/mL (mean +/- SD) (range, 65-143) after 60 weeks, 121.63 +/- 27.85 pg/mL (mean +/- SD) (range, 62-179) after 100 weeks, 63 +/- 19.85 pg/mL (mean +/- SD) (range, 68-723) after 150 weeks and 102 +/- 18.65 pg/mL (mean +/- SD) (range, 68-113) after 200 weeks. The prevalence of hypoparathyroidism (serum iPTH level of < 20 pg/mL with a normal or low serum calcium concentration) was 2 out of 35 patients (5.71%) by week 60, with recovery of normal values by week 100. Graft-related recurrence occurred in one out of 35 patients (2.85%). CONCLUSIONS: SCPTx after TPT and PTx for secondary RHP is an adequate method to replace muscular forearm parathyroid transplantation and avoid its complications. The functional results of TPT and SCPTx compare favorably with published data on other surgical techniques proposed for the treatment of RHP. Long-term follow-up of this series is currently being performed. PMID- 17916287 TI - [Laparoscopic adrenalectomy]. AB - INTRODUCTION: In the last few years, laparoscopic adrenalectomy has become widely used in the management of adrenal disease. MATERIAL AND METHOD: We reviewed our experience of 24 patients who underwent laparoscopic adrenalectomy between 1998 and 2006. RESULTS: Surgery was indicated for Cushing's syndrome in 46% of the patients, aldosteronoma in 25%, incidentaloma in 21% and pheochromocytoma in 8%. A lateral transabdominal approach was employed in all patients. The mean age of the patients was 50.4 years (17 women and 7 men). Left unilateral adrenalectomy was performed in 63% of the patients, right unilateral adrenalectomy in 29% and bilateral adrenalectomy in 8%. The conversion rate was 4%. The mean operating time was 134 minutes in unilateral approaches and 245 minutes in bilateral approaches. The mean size of the gland was 4 cm. The complications rate was 4% and there was no mortality. Disease control was achieved in 96% of the patients after a mean follow-up of 49 months. CONCLUSIONS: Laparoscopic adrenalectomy should be considered the procedure of choice for the surgical management of benign adrenal disease. PMID- 17916288 TI - [Effect of neoadjuvant chemotherapy on the results of resection of colorectal liver metastases]. AB - INTRODUCTION: Surgery is the treatment of choice in patients with colorectal liver metastases. However, only 10% to 20% of these cases are resectable. The use of neoadjuvant chemotherapy may allow surgery in patients with tumors initially considered unresectable. The aim of this study was to compare the results of liver resection due to colorectal liver metastases in patients with and without neoadjuvant chemotherapy. PATIENTS AND METHOD: We studied 105 patients who underwent surgery for liver metastases from colorectal cancer. The patients were divided into two groups according to treatment: surgery in patients with initially resectable tumors (group 1) and neoadjuvant chemotherapy plus surgery (group 2) in patients with initially irresectable tumors, who were considered for surgery after response to chemotherapy. Age, sex, origin of primary tumor, time of presentation, number, maximum size and location of metastases, CEA, resection margin, postoperative morbidity and mortality, length of hospital stay, recurrence rate, survival and disease-free survival were compared between the 2 groups of patients. RESULTS: When group 1 was compared with group 2, statistically significant differences were observed in synchronicity (30.8% vs 77.4%), bilobarity (13.5% vs 58.5%), number and size of metastases (1 vs 3 nodules and 4 cm vs 2 cm), resectability rate (96.1% vs 81.1%), disease-free interval (25 vs 11 months) and long-term survival at 1, 3 and 5 years (93%, 67% and 36% vs 78%, 26% and 12%). However, no statistically significant differences were found in postoperative morbidity and mortality (28.8% and 0% in group 1 and 22.6% and 1.8% in group 2, respectively). CONCLUSIONS: Neoadjuvant chemotherapy was not associated with greater postoperative morbidity and mortality after resection of colorectal liver metastases, but long-term survival was lower in the group of patients receiving this treatment modality than in those with tumors initially considered resectable. PMID- 17916289 TI - [Retroperitoneal sarcomas. Report of five new cases and review of the current situation]. AB - INTRODUCTION: Soft-tissue sarcomas represent a heterogeneous group of rare tumors arising from the mesenchymal cells of the connective tissue. Approximately 15% of these tumors arise in the retroperitoneum. These neoplasms are locally aggressive and the only curative treatment is surgical resection "en-bloc". The main cause of mortality is locoregional recurrence. Five-year survival is 3-58%, depending on the histologic subtype and grade. PATIENTS AND METHOD: Over a 5-year period (2001 to 2006), we performed surgery in five patients with retroperitoneal sarcomas. The mean age was 59 years (range, 46-76) with a male-to-female ratio of 3:2. The most frequent signs were abdominal pain and the appearance of a mass. In 2 patients, an incidental diagnosis was made during the surgical intervention. RESULTS: Surgical resection was performed in 5 patients but was incomplete in two patients. We removed 2 liposarcomas, 1 leiomyosarcoma, 1 chondrosarcoma and 1 fusocellular sarcoma arising in the kidney. Locoregional recurrence occurred in four patients, requiring between 1 and 2 new relaparotomies (using the retroperitoneal approach in 2 patients). There was one death. CONCLUSIONS: Retroperitoneal sarcomas are a heterogeneous group of tumors, in which the only common factor is the anatomical location. New randomized, prospective, multicenter trials are required to apply different therapeutic options according to the subtype of retroperitoneal sarcoma. PMID- 17916290 TI - [Giant hepatic echinococcus cyst with cysto-pleural fistula and pleural echinococcosis]. AB - Extrahepatic complications from hepatic cystic echinococcosis are rare and may be life threatening. Although the prevalence of echinococcosis in Spain had decreased, the number of cases of this disease and its severity has risen again due to immigration. We report the case of a patient with a giant hepatic echinococcus cyst diagnosed during investigation of an abdominal mass. The mass was associated with three other cysts: the first cyst was fistulized to the biliary tract, without clinical or laboratory alterations, the second cyst was fistulized to the right pleural cavity, leading to pleural echinococcosis and respiratory distress due to massive pleural effusion, and the third cyst was calcified. The challenge in this case layed in its diagnosis and treatment. The clinical presentation was unusual due to the extension of the disease, hampering complete cystic excision. PMID- 17916291 TI - [Gallstones in a retained gastric antrum]. AB - Retained gastric antrum arises when there is incomplete excision of the gastric antrum during Billroth II gastrectomy for peptic ulcer disease. We report the case of a patient with gallstones in a retained gastric antrum, without biliodigestive fistula. This finding is extremely rare and we have found no previously reported cases in the literature. PMID- 17916292 TI - [Intrahepatic cholangiocarcinoma on a hydatidic cyst]. AB - Cholangiocarcinomas are malignant tumors arising from the bile ducts. One of the factors associated with the development of malignancy is parasitic infections by helminths trematodes that cause liver fluke infections. This association is very common in south-east Asia, but association with liver hydatidosis has not been reported in the scientific literature. We report a case in which these two diseases were associated and required non-radical surgical treatment, since radiological findings were not suspicious for these entities. PMID- 17916293 TI - [Pneumoretroperitoneum]. PMID- 17916295 TI - [Small is better]. PMID- 17916294 TI - [Transgastric percutaneous drainage of pancreatic pseudocysts: also of use in traumatic pseudocysts]. PMID- 17916296 TI - [Survival above 5-years in malignant peritoneal mesothelioma]. PMID- 17916297 TI - [Hemorrhagic shock due to spontaneous rupture of a hepatocellular carcinoma]. PMID- 17916298 TI - [Atypical pancreatic somatostatinoma]. PMID- 17916299 TI - [The law on effective equality between men and women: a step forward]. PMID- 17916300 TI - [The advertising message of drugs advertisements in Spanish medical journals]. AB - OBJECTIVE: To describe the characteristics of advertising in Spanish medical journals and to analyze the advertising message in drug advertisements. METHODS: Six issues of 4 Spanish medical journals published in 2001 were reviewed to identify the number of advertisements and their characteristics. The journals selected were Atencion Primaria, Anales Espanoles de Pediatria, Medicina Clinica and Gaceta Sanitaria. The advertising message was analyzed by evaluating 5 factors: communication aim, sales argument, communication treatment, and use of text and image. RESULTS: 609 advertisements were found. Drug advertisements were the most numerous (69.9%). Advertising pressure was highest in Atencion Primaria (36%), followed by Anales Espanoles de Pediatria (22%), Medicina Clinica (12%) and Gaceta Sanitaria (4%). Of the 195 drug advertisements analyzed, the aim of the advertising message was mostly to present or remind readers of an existing product (70.8%). The sales argument was rational in 86.5%. The communication treatment was advertising in 72.6%. The text of the advertisement contained a headline in 82.4% or a slogan in 50.8%. The advertisement image was a photograph in 74.7% and the aim of the image was to present the product (48.7%), to promise a benefit (45.1%), or to argue its qualities (31.1%). CONCLUSIONS: In the journals aimed at prescribing physicians advertising pressure was higher and advertisements were intercalated in article text. Advertising concerned already existing products, used rational arguments, and the communication treatment was advertising. PMID- 17916301 TI - [Trends in social inequalities in pregnancy care in Barcelona (Spain), 1994-97 versus 2000-03]. AB - OBJECTIVE: To describe and compare social inequalities in pregnancy care among pregnant women living in Barcelona (Spain) in 2 periods. METHODS: Two 4-year periods were compared: 1994-1997 and 2000-2003. The study population consisted of pregnant women living in Barcelona and the control sample was drawn from the Barcelona Birth Defects Registry (n = 905 in 1994-1997; n = 927 in 2000-2003). Medical records and personal interviews with the mothers were used as information sources. The dependent variables were pregnancy planning, prenatal use of folic acid, smoking, the number of obstetric visits, trimester of the first visit, the number of obstetric ultrasound scans, fifth-month diagnostic ultrasound scan, invasive procedures, and smoking cessation. The independent variables were maternal age and social class. Maternal age-adjusted logistic regression models for each dependent variable according to social class were calculated and the results for both 4-year periods were compared. RESULTS: Pregnant women in both manual and non-manual occupational classes showed better results in the second period in 7 out of 10 variables (although the results were not identical in the 2 occupational classes). However, when interclass variations between the 2 periods were compared, differences in 8 out of 10 variables were found: 7 indicators were more favorable in the more privileged classes and only one was more favorable in the less privileged classes. CONCLUSIONS: Except for one of the variables analyzed (more than 3 ultrasound scans), the less privileged classes showed poorer results than the more privileged classes when the tendencies in indicators were compared between the two periods. The gap between social classes in pregnancy care is increasing over time. PMID- 17916302 TI - [Changes in physicians' attitudes to computerized ambulatory medical record systems: a longitudinal qualitative study]. AB - OBJECTIVES: To explore physicians' beliefs about a computerized ambulatory medical record system at different stages of its implementation. METHODS: We performed a longitudinal qualitative in-depth interview study (July 2001 to December 2003) in the Hospital Italiano, Buenos Aires, Argentina. Semi-structured interviews were conducted in 20 primary care cardiologists purposively selected before, during and after the system's implementation process (10 interviews per stage). The interviews were independently analyzed by 2 researchers, who jointly designed an agreed category list. RESULTS: Both before and during the first stage of the implementation process, the physicians expected that that the system would improve healthcare-related administration and increase accessibility to individual data. However, they did not foresee that the system's shared information could modify the clinical aspects of patient care. By the end of the implementation process, the physicians realized that the system provided them with a broader perspective on their patients, which in turn improved their own professional performance. Throughout the implementation, the physicians were against using the computer while the patient was present. This opposition prevented them from regarding the system as part of the medical consultation and from considering data from the system as direct patient-related signs. CONCLUSIONS: The system's implementation modified the physicians' views on computerized ambulatory medical records, as they eventually considered them as an ancillary tool to clinical activity. The value assigned to the system depends on its relevance within the institutional framework. PMID- 17916303 TI - [Descriptive epidemiology of non-meningococcal bacterial meningitis in the province of Saragossa [Spain] from 1999 to 2004. Evaluation of the Epidemiological Surveillance System]. AB - OBJECTIVES: To describe non-meningococcal bacterial meningitis (nMM) and to evaluate the Epidemiological Surveillance System (ESS) in the province of Saragossa (Spain) between 1999 and 2004. METHODS: Information was obtained from the register of diseases subject to mandatory reporting and the Minimum Basic Data Set (MBDS). The ESS was evaluated by using the Centers for Disease Control and Prevention (CDC) criteria and by estimating the completeness of the system through the capture-recapture technique. RESULTS: 111 cases of nMM were notified (62.2% in males) and the mean age was 40.7 years. The largest proportion of cases (16.5%) occurred in children under 2 years of age. The clinical presentation was meningitis in 81.1%. Diagnosis was through bacterial culture in 70.3%. Streptococcus sp. was found in 54% (82% due to S. pneumoniae), enterobacteria in 5.4%, Listeria and Staphylococcus in 4.5%, Pseudomonas aeruginosa in 1.8%, and Haemophilus influenzae in 0.9%. The incidence rate per 100,000 inhabitants was 1.6 in 2004 and 2.6 in 2001. The case fatality was 7.3%. The completeness of the mandatory reporting system reached a peak in 2004 at 84.4%. The combined completeness of the MBDS and the mandatory reporting system was above 85% and timeliness of reporting was 2 days. The acceptability of the system was high since 75% of all variables were complete in 97% of the questionnaires. CONCLUSION: We highlight the importance of evaluation of the ESS, based on its results. nMM due to S. pneumoniae represent an important group of diseases and their case fatality is high. The completeness of the ESS in Saragossa was over 80% when the mandatory reporting system and the MBDS were combined. Incorporating the MBDS into surveillance would facilitate the estimation of the real incidence of various diseases subject to mandatory reporting. PMID- 17916304 TI - [Two methods to analyze trends in the incidence of heroin and cocaine use in Barcelona [Spain]]. AB - OBJECTIVES: To describe 2 statistical methods for estimating trends in the incidence of heroin and cocaine use in Barcelona. METHODS: Admissions for treatment of heroin and cocaine consumption recorded by the Barcelona Drug Information System between 1991 and 2003 were used. We selected 4,367 subjects initiating treatment for the first time for heroin use, and 2,147 for cocaine use. Two statistical techniques were employed: Reporting Delay Adjustment (RDA) and the Log-linear Model (LLM). RDA was used in subjects who initiated drug consumption between 1991 and 2003, and LLM for those who began heroin use between 1967 and 2003 and cocaine use between 1971 and 2003. In addition, for each drug and method the latency period (LP) was determined (years between first consumption and first treatment). RESULTS: Comparison of the distributions of the LP for each drug revealed that heroin users initiated treatment for the first time sooner than cocaine users, regardless of the method employed. In general, the estimated incidence of heroin use in Barcelona fell progressively after 1982. In contrast, the incidence of cocaine use rose rapidly until 1998, and has been irregular since. The incidence of cocaine use began to be substantial in the early 1990s, but took several years to manifest itself as problematic. CONCLUSION: The estimated incidence was underestimated by RDA compared with LLM, but the incidence of heroin use could be biased before 1991 due to changes in treatment provisions. Although the estimated incidence is relative to individuals who are admitted for treatment at some time in their life, trends in incidence can be used to plan future actions. PMID- 17916305 TI - [Differences between prescription and dispensing of generic medicines in primary care. When the exception is the rule]. AB - OBJECTIVES: To quantify the frequency and cost of switching prescribed generic brand drugs for others at dispensation in pharmacies. METHODS: We performed a descriptive cross-sectional study to compare prescription and dispensation of generic drugs in a primary care quota by using computerized data from medical consulting rooms and the Pharmaceutical Service Management Program in 2005. RESULTS: Of 16,181 prescriptions, 3,759 generic prescriptions were included in the study, with a mean price of 7.61 euros as opposed to 8.01 euros for those dispensed in the retail pharmacy. In 62.9% of drugs, the brand prescribed by the doctor was not dispensed. In 65.1% of cases in which the brand was switched, the drug was changed for another generic drug with a higher price. CONCLUSIONS: Changing the prescribed generic drug was common in the quota studied with a tendency to dispense another drug that was more expensive, thereby increasing costs. PMID- 17916306 TI - [Comment. Substitution at pharmacies and generic market]. PMID- 17916307 TI - Scientific production in bioethics in Spain through MEDLINE. AB - OBJECTIVE: To describe Spain's scientific production in the field of bioethics from 1966 to 2003. METHODS: Manuscripts published by Spanish authors between 1966 and 2003 and containing key word references to bioethics, ethics, and 22 other related terms were retrieved from the Medline database. RESULTS: 858 documents were selected: 78 (9.1%) were published between 1966 and 1983, 163 (19%) between 1984 and 1993, and 617 (71.9%) between 1994 and 2003. The main subject areas treated were laws and rights (15.4%) and research and ethics committees (13.1%). The last of these periods witnessed an increase in publications on genetics and human cloning and a decrease in those treating abortion. Institutional affiliations referred mainly to universities (38.9%) and hospitals (38.5%). CONCLUSIONS: There was a progressive increase in the number of scientific publications on bioethics by Spanish authors during the study period. PMID- 17916308 TI - [A survey of gastroprotective drugs: prescription-indication in hospitalized patients]. AB - Prescription rates of anti-ulcer drugs in hospitals and their spill-over to general practice have risen over the last few years, increasing pharmaceutical expenses. The aim of this study was to analyze gastroprotective drug prescription habits in our hospital by assessing both prevalence and adherence to approved indications. An observational study of prescription-indication was performed with 2 cross sections separated by 6 months to avoid selection bias. We found overprescription of gastroprotective drugs, mainly proton pump inhibitors for the prevention of non-steroidal antiinflammatory drug-induced ulcer (17.1%). Overall, 77.6% of prescriptions had no acceptable indication, despite the availability of a specific protocol produced by the Pharmacy and Therapeutics Committee in our center. There was a high prevalence of prescriptions for non-approved indications such as prophylaxis in patients administered corticosteroids without non steroidal anti-inflammatory agents (25.7%). New programs to train clinicians and clinical managers in rational drug use are required. PMID- 17916309 TI - [Statistics and public health: the argument of the numerical method]. PMID- 17916310 TI - [Influence of big cities' lifestyle on the health and longevity of their population: study applied to our city. 1888]. PMID- 17916311 TI - [Systematic review of studies on the socioeconomic status of men who batter their intimate partners]. AB - BACKGROUND: Despite the visibility of intimate partner violence against women in low socioeconomic groups, the association of low socioeconomic status and violent behavior is unclear. We performed a systematic review of the empirical evidence on the causal role of batterers' socioeconomic status in this phenomenon. METHODS: We performed a systematic review, using the following databases: Eric (1966-2004), Sociological Abstracts (1963-2005), Science Citation Index (1945 2005), Social Science Citation Index (1956-2005), Medline through Pubmed (1966 2005), Social Service Abstracts (1980-2005), Lilacs (1982-2005) and Psycinfo (1972-2005). We included empirical papers with aims or hypothesis related to the causal relation between low socioeconomic status (employment, education, income) in men and IPV. RESULTS: A total of 251 studies were identified, but only 10 met the inclusion criteria. There was one cohort study, one case-control study, one ecological study and one study based on a series of cases. Two studies calculated odds ratio in the analyses. One of these studies reported an odds ratio of 1.4 (95% CI, 1.1-1.9) with unemployment and the other study showed no significant positive associations with this factor or with low income or education. CONCLUSIONS: More information and better quality data are required to establish conclusive results on the causal role of the socioeconomic status of men who batter their intimate partners. The empirical evidence on the relationship between violent male behavior against their partners and low socioeconomic status is still insufficient. PMID- 17916313 TI - [Sanitary progress = more? Or less and better?]. PMID- 17916312 TI - [Is racism an important issue in public health in Spain?]. PMID- 17916314 TI - Intravitreal bevacizumab therapy for neovascular age-related macular degeneration with large submacular hemorrhage. AB - PURPOSE: To evaluate functional and anatomic effects of intravitreal bevacizumab (Avastin; Roche Pharma, Vienna, Austria) in patients with neovascular age-related macular degeneration (AMD) with large submacular hemorrhages. DESIGN: Retrospective, clinical study. METHODS: Twenty-one eyes of 19 AMD patients with choroidal neovascularization and large submacular hemorrhage involving the fovea comprising more than 50% of the total lesion area were evaluated. All patients completed at least four months of follow-up; 12 patients fulfilled 12 months or more of follow-up. Patients were treated with up to six intravitreal bevacizumab injections (1 mg/0.04 ml) at a minimum of four-week intervals. Changes from baseline visual acuity (VA) scores, retinal measurements by optical coherence tomography (OCT), angiographic lesion characteristics, and hemorrhage size were analyzed. A safety assessment was performed at all visits. RESULTS: Intravitreal bevacizumab injections were well tolerated in all patients. At month 4, VA was stable or improved (visual loss of 3 acuity lines or fewer) in 100% and improved by at least 3 lines in 9.5%. Comparable results were found at month 12. On average, the central foveal thickness decreased significantly by 55 microm four weeks after the first injection (P < .001) and by 52 microm at month 4 (P = .002). A significant anatomic improvement also was found for maximum retinal thickness, minimum retinal thickness, and foveal volume (P < .05) and was maintained during four months of follow-up. Mean size of hemorrhage was significantly reduced from 19.7 mm(2) at baseline to 2.5 mm(2) at the four-month follow-up (P < .001). CONCLUSIONS: Intravitreal bevacizumab seems to be a promising therapeutic option in eyes with neovascular AMD and large submacular hemorrhages, with a stabilization in VA and anatomic improvement. PMID- 17916315 TI - Retinal detachment after laser in situ keratomileusis in myopic eyes. AB - PURPOSE: To investigate the incidence, characteristics, and surgical management of retinal detachment (RD) after laser in situ keratomileusis (LASIK) surgery in myopia. DESIGN: Retrospective, observational case series. METHODS: We retrospectively reviewed the RDs observed in 18,342 eyes (9,598 patients) that underwent LASIK for the correction of myopia. All patients had no history of corneal diseases, and preoperative examinations, including a thorough fundus examination, were performed. Patients were followed up for a mean of 20 months (range, four to 27 months), and the clinical features of the RD eyes after LASIK were investigated. RESULTS: RD developed in six patients, including two males and four females. The incidence of a RD after LASIK in this study was 0.033%. Mean degree of pre-LASIK myopia in these eyes was -9.33 diopters (D; range, -6.25 to 14.00 D). The mean interval between LASIK and RD development was 9.25 months (range, two to 18 months). All RDs occurred spontaneously and were managed with vitreoretinal surgeries. Retinal reattachment was achieved at the first RD surgery in all six eyes (100%) at a mean follow-up of 9.3 months (range, three to 18 months). CONCLUSIONS: RD after LASIK for correction of myopia is uncommon. This study suggested no cause-and-effect relationship could be proven between RD development and LASIK procedure in myopia. Clinicians should still be aware of retinal pathologic features in patients undergoing LASIK. PMID- 17916316 TI - Octreotide long-acting repeatable for the treatment of chronic macular edema in uveitis. AB - PURPOSE: To report on the efficacy of the somatostatin analog octreotide long acting repeatable (LAR), in the treatment of uveitic chronic macular edema (CME). DESIGN: Case series, retrospective analysis. METHODS: In 20 patients, 20 episodes of recurrent CME during otherwise quiescent uveitis were treated with intramuscular octreotide LAR injections. Patients were included if CME control with acetazolamide or systemic and periocular steroids had failed during previous CME episodes or if contraindications existed for persistent use of these therapies. Mean outcome points were CME and visual acuity changes. Correlation of prognostic factors with these outcomes was analyzed. RESULTS: The included CME episodes occurred 7.6 +/- 1.4 years after onset of uveitis. Octreotide LAR treatment started 7.0 +/- 7.3 months after diagnosis of CME. CME decreased in 70% of episodes, after 2.7 +/- 1.3 months of treatment. After arrest of successful treatment, CME recurred instantly (27.2%) or within six months (36.4%). In 36.4% of successfully treated episodes, CME was absent for more than one year. A probable prognostic factor for success was the duration of CME before treatment. CONCLUSIONS: Octreotide LAR had an edema-reducing effect in 70% of treated CME episodes. Successful response was related to duration of CME before start of treatment. The early recurrence of CME (63.6%) after arrest of octreotide LAR advocates a long-term treatment in recent episodes of macular edema in otherwise quiescent uveitis. PMID- 17916317 TI - The impact of nasal conjunctivochalasis on tear functions and ocular surface findings. AB - PURPOSE: To investigate the effect of grade 3 nasal conjunctivochalasis (NCCh) on the tear film inflammation, ocular surface findings, and tear function findings. DESIGN: Prospective, observational case series. METHODS: Eleven eyes with Yokoi grade 3 NCCh in which the chalatic conjunctiva occluded the punctum and 18 eyes without NCCh but with central or temporal conjunctivochalasis, or both, and 16 eyes of healthy controls were recruited prospectively. Enzyme-linked immunosorbent assay for inflammatory tear cytokines, tear film break-up time (BUT), Schirmer I test measurements, and fluorescein and rose bengal vital stainings and impression cytologic and brush cytologic analysis for real-time reverse-transcriptase polymerase chain reaction analysis of MUC5AC messenger ribonucleic acid (mRNA) expression were performed. RESULTS: Eyes with grade 3 NCCh had significantly delayed tear clearance. All inflammatory cytokines showed higher values in eyes with grade 3 NCCh compared with the eyes without nasal chalasis with a comparably significant elevation in interleukin-1b and tumor necrosis factor alpha levels. The mean rose bengal score in eyes with grade 3 NCCh was significantly higher compared with eyes without nasal chalasis and eyes of controls. The mean goblet cell density was significantly lower in eyes with grade 3 NCCh with downregulation of the relative MUC5AC mRNA expression. CONCLUSIONS: Inflammation plays an important role in the pathogenesis of conjunctivochalasis and is more pronounced in eyes with nasal chalasis. Pooling of inflammatory cytokines in tears of patients with NCCh associated with delayed tear clearance induces distinct adverse effects that affect the ocular surface health. PMID- 17916318 TI - Long-term outcomes of Ahmed glaucoma valve implantation in refractory glaucomas. AB - PURPOSE: To evaluate the long-term efficacy of intraocular pressure (IOP) reduction and complications of Ahmed glaucoma valve (AGV) implantation (New World Medical, Inc, Rancho Cucamonga, California, USA) in refractory glaucoma. DESIGN: Retrospective cohort study. METHODS: Retrospective medical records from 64 patients (78 eyes) with refractory glaucoma who underwent AGV implantation with a minimum of three years of follow-up were reviewed. Data regarding age, gender, race, eye laterality, specific glaucoma diagnosis, best-corrected visual acuity (BCVA), number of medications, IOP, visual fields, surgical complications, and follow-up interval were collected from all visits and were analyzed. The primary outcome measure was cumulative probability of success defined as IOP of less than 21 mm Hg and of 5 mm Hg or more with a minimum of 15% reduction from baseline IOP, without additional glaucoma surgery or loss of light perception. Secondary outcomes included IOP and number of medications at three, six, 12, 24, 36, 48, and 60 months after surgery, surgical complications, and final BCVA. RESULTS: The cumulative probability of success was 80% and 49% at one and five years, respectively. IOP was reduced from a mean of 30.4 +/- 10.7 mm Hg to 17.0 +/- 5.0 mm Hg at 12 months and 15.9 +/- 3.0 mm Hg at 60 months (P < .001). The number of medications decreased from 3.2 +/- 1.0 medications at baseline to 1.6 +/- 0.4 at 12 months and 2.1 +/- 0.2 at 60 months (P < .001). Prior glaucoma surgery and the silicone type of AGV were statistically significant risk factors for failure (P < .001). CONCLUSIONS: Approximately 50% of single-plate AGV implantations in refractory glaucoma were considered successful after five years of follow-up. Prior glaucoma surgery was a statistically significant risk factor for failure. PMID- 17916319 TI - Central retinal vein occlusion case-control study. AB - PURPOSE: To investigate risk factors for central retinal vein occlusion (CRVO). DESIGN: Retrospective case-control study. METHODS: Consecutive patients with CRVO examined from July 1, 2005 through July 31, 2006 were compared with an historical gender- and age-matched control group of patients with ocular problems other than vascular occlusive disease from the same referral practice. Risk factors for CRVO were evaluated. RESULTS: The 144 patients in the CRVO group, 87 males and 57 females, had a mean age of 69.6 years (+/-13.6 years). CRVO was associated with hypertension (P < .001), diabetes mellitus (P = .047), glaucoma (P < .001), atrial fibrillation (P = .036), angiotensin-converting enzyme inhibitor use (P = .022), aspirin use (P < .001), and warfarin use (P = .011) by univariate analyses. Postmenopausal estrogen use was more common among women in the control group (P = .029). Multivariate logistic regression found the independent predictors for CRVO to be: glaucoma (adjusted odds ratio [OR], 4.75; P < .001), aspirin use (adjusted OR, 2.66; P = .001), and warfarin use (adjusted OR, 3.34; P = .005). CONCLUSIONS: We found many of the same risk factors previously identified for CRVO by other studies, but we identified both aspirin and warfarin use to be independent risk factors for CRVO. Although these findings suggest the vasculopathic and prothrombotic risks in some patients may not be addressed adequately by antithrombotic therapy, they also suggest that the pathogenesis of CRVO may be more complicated than just the development of a primary thrombus within the vein. PMID- 17916320 TI - Early bevacizumab treatment of central retinal vein occlusion. AB - PURPOSE: To evaluate the change in visual acuity and retinal appearance in patients after early initiation of intravitreal bevacizumab treatment for central retinal vein occlusion (CRVO). DESIGN: Retrospective, interventional case series. METHODS: Patients with CRVO of fewer than three months' duration receiving intravitreal bevacizumab as primary treatment were evaluated. Patients received an intravitreal 1.25 mg (0.05 ml) bevacizumab injection. Changes in visual acuity, central macular thickness, venous tortuosity and diameter, and optic disk edema were noted. RESULTS: Six eyes of five consecutive patients with CRVO treated with intravitreal bevacizumab injection were reviewed retrospectively. The patients did not have other ocular conditions that could have compromised visual acuity. The mean baseline visual acuity was 20/428 (logarithm of the minimum angle of resolution [logMAR] units, 1.33). The mean follow-up period was 12 months (range, seven to 15 months), and the number of bevacizumab injections ranged from four to 10. The patients showed a statistically significant decrease in optic nerve head swelling, venous tortuosity, and venous diameter, with the largest proportion of change occurring within one month of the first bevacizumab injection. The mean visual acuity at last follow-up was 20/53 (logMAR units, 0.42; P = .035, as compared with baseline). In no patient did collateral vessels at the optic nerve head develop. CONCLUSIONS: The patients experienced a dramatic improvement in the visual acuity and clinical fundus appearance, without collateral vessel formation. These findings are difficult to explain with current theories of the pathophysiologic features of CRVO. These findings also suggest early initiation of anti-vascular endothelial growth factor (VEGF) treatment should be studied in a larger trial for CRVO. PMID- 17916321 TI - Transmembrane proteins of tight junctions. AB - Tight junctions contribute to the paracellular barrier, the fence dividing plasma membranes, and signal transduction, acting as a multifunctional complex in vertebrate epithelial and endothelial cells. The identification and characterization of the transmembrane proteins of tight junctions, claudins, junctional adhesion molecules (JAMs), occludin and tricellulin, have led to insights into the molecular nature of tight junctions. We provide an overview of recent progress in studies on these proteins and highlight their roles and regulation, as well as their functional significance in human diseases. PMID- 17916322 TI - 1-Alkanols and membranes: a story of attraction. AB - Although 1-alkanols have long been known to act as penetration enhancers and anesthetics, the mode of operation is not yet understood. In this study, long time molecular dynamics simulations have been performed to investigate the effect of 1-alkanols of various carbon chain lengths onto the structure and dynamics of dimyristoylphosphatidylcholine bilayers. The simulations were complemented by microcalorimetry, continuous bleaching and film balance experiments. In the simulations, all investigated 1-alkanols assembled inside the lipid bilayer within tens of nanoseconds. Their hydroxyl groups bound preferentially to the lipid carbonyl group and the hydrocarbon chains stretched into the hydrophobic core of the bilayer. Both molecular dynamics simulations and experiments showed that all 1-alkanols drastically affected the bilayer properties. Insertion of long-chain 1-alkanols decreased the area per lipid while increasing the thickness of the bilayer and the order of the lipids. The bilayer elasticity was reduced and the diffusive motion of the lipids within the bilayer plane was suppressed. On the other hand, integration of ethanol into the bilayer enlarged the area per lipid. The bilayer became softer and lipid diffusion was enhanced. PMID- 17916323 TI - The gamma-core motif correlates with antimicrobial activity in cysteine containing kaliocin-1 originating from transferrins. AB - Kaliocin-1 is a 31-residue peptide derived from human lactoferrin, and with antimicrobial properties that recapitulate those of its 611 amino acid parent holoprotein. As kaliocin-1 is a cysteine-stabilized peptide, it was of interest to determine whether it contained a multidimensional gamma-core signature recently identified as common to virtually all classes of disulfide-stabilized antimicrobial peptides. Importantly, sequence and structural analyses identified an iteration of this multidimensional antimicrobial signature in kaliocin-1. Further, the gamma-core motif was found to be highly conserved in the transferrin family of proteins across the phylogenetic spectrum. Previous studies suggested that the mechanism by which kaliocin-1 exerts anti-candidal efficacy depends on mitochondrial perturbation without cell membrane permeabilization. Interestingly, results of a yeast two-hybrid screening analysis identified an interaction between kaliocin-1 and mitochondrial initiation factor 2 in a Saccharomyces cerevisiae model system. Taken together, these data extend the repertoire of antimicrobial peptides that contain gamma-core motifs, and suggest that the motif is conserved within large native as well as antimicrobial peptide subcomponents of transferrin family proteins. Finally, these results substantiate the hypothesis that antimicrobial activity associated with host defense effector proteins containing a gamma-core motif may correspond to targets common to fungal mitochondria or their bacterial ancestors. PMID- 17916324 TI - Ontogeny up-regulates renal Na(+)/Cl(-)/creatine transporter in rat. AB - Creatine plays a role in energy storage and transport/shuttle of high-energy phosphate in heart, brain, retina, testis and skeletal muscle. These tissues take creatine from the plasma via a 2Na(+)/1Cl(-)/1creatine cotransporter (CRT). We have previously demonstrated that renal apical membrane presents a 2Na(+)/1Cl( )/1creatine cotransport activity. The goal of this study was to determine whether this transporter is ontogenically regulated. Na(+)/Cl(-)/creatine transport activity was evaluated by measuring [(14)C]-creatine uptake into renal brush border membrane vesicles. CRT mRNA expression was measured by Northern and real time PCR assays. E20 foetuses, newborn, suckling, weaning and adult (2- and 8 month-old) Wistar rats were used. The results revealed that neither the vesicular volume, the binding of creatine to the brush-border membrane vesicles, nor the purity of the brush-border membrane vesicle preparations was affected by maturation. Fetal and neonatal kidneys contained a creatine transporter that was qualitatively indistinguishable from that in the adult: it was concentrative, Na(+)- and Cl(-)-dependent, electrogenic and inhibited by guanidinopropionic acid. Maturation increased this transport activity by increasing the maximal rate of transport (V(max)) without significantly changing the apparent K(m). Northern analysis revealed two transcripts for CRT of 2.7 kb and 4.2 kb in all the ages tested. Northern and real-time PCR assays showed that, as seen with NaCl dependent creatine transport activity, maturation increased CRT mRNA expression. This study reports for the first time that: (i) an apical renal Na(+)/Cl( )/creatine cotransporter is already active in rat foetuses and (ii) development regulates Na(+)/Cl(-)/creatine cotransport activity by increasing the density and/or turnover of the transporters. PMID- 17916325 TI - NMR structural studies of the bacterial outer membrane protein OmpX in oriented lipid bilayer membranes. AB - The beta-barrels found in the outer membranes of prokaryotic and eukaryotic organisms constitute an important functional class of proteins. Here we present solid-state NMR spectra of the bacterial outer membrane protein OmpX in oriented lipid bilayer membranes. We show that OmpX is folded in both glass-supported oriented lipid bilayers and in lipid bicelles that can be magnetically oriented with the membrane plane parallel or perpendicular to the direction of the magnetic field. The presence of resolved peaks in these spectra demonstrates that OmpX undergoes rotational diffusion around an axis perpendicular to the membrane surface. A tightly hydrogen-bonded domain of OmpX resists exchange with D2O for days and is assigned to the transmembrane beta-barrel, while peaks at isotropic resonance frequencies that disappear rapidly in D2O are assigned to the extracellular and periplasmic loops. The two-dimensional 1H/15N separated local field spectra of OmpX have several resolved peaks, and agree well with the spectra calculated from the crystal structure of OmpX rotated with the barrel axis nearly parallel (5 degrees tilt) to the direction of the magnetic field. The data indicate that it will be possible to obtain site-specific resonance assignments and to determine the structure, tilt, and rotation of OmpX in membranes using the solid-state NMR methods that are currently being applied to alpha-helical membrane proteins. PMID- 17916326 TI - Membrane interactions of hemoglobin variants, HbA, HbE, HbF and globin subunits of HbA: effects of aminophospholipids and cholesterol. AB - The interaction of hemoglobin with phospholipid bilayer vesicles (liposomes) has been analyzed in several studies to better understand membrane-protein interactions. However, not much is known on hemoglobin interactions with the aminophospholipids, predominantly localized in the inner leaflet of erythrocytes, e.g., phosphatidylserine (PS), phosphatidylethanolamine (PE) in membranes containing phosphatidylcholine (PC). Effects of cholesterol, largely abundant in erythrocytes, have also not been studied in great details in earlier studies. This work therefore describes the study of the interactions of different hemoglobin variants HbA, HbE and HbF and the globin subunits of HbA with the two aminophospholipids in the presence and absence of cholesterol. Absorption measurements indicate preferential oxidative interaction of HbE and alpha-globin subunit with unilamellar vesicles containing PE and PS compared to normal HbA. Cholesterol was found to stabilize such oxidative interactions in membranes containing both the aminophospholipids. HbE and alpha-globin subunits were also found to induce greater leakage of membrane entrapped carboxyfluorescein (CF) using fluorescence measurements. HbE was found to induce fusion of membrane vesicles containing cholesterol and PE when observed under electron microscope. Taken together, these findings might be helpful in understanding the oxidative stress-related mechanism(s) involved in the premature destruction of erythrocytes in peripheral blood, implicated in the hemoglobin disorder, HbE/beta-thalassemia. PMID- 17916327 TI - Structural biology of bacterial iron uptake. AB - To fulfill their nutritional requirement for iron, bacteria utilize various iron sources which include the host proteins transferrin and lactoferrin, heme, and low molecular weight iron chelators termed siderophores. The iron sources are transported into the Gram-negative bacterial cell via specific uptake pathways which include an outer membrane receptor, a periplasmic binding protein (PBP), and an inner membrane ATP-binding cassette (ABC) transporter. Over the past two decades, structures for the proteins involved in bacterial iron uptake have not only been solved, but their functions have begun to be understood at the molecular level. However, the elucidation of the three dimensional structures of all components of the iron uptake pathways is currently limited. Despite the low sequence homology between different bacterial species, the available three dimensional structures of homologous proteins are strikingly similar. Examination of the current three-dimensional structures of the outer membrane receptors, PBPs, and ABC transporters provides an overview of the structural biology of iron uptake in bacteria. PMID- 17916328 TI - The neural circuitry mediating shifts in behavioral response and cognitive set in autism. AB - BACKGROUND: Recent studies have suggested that the social and cognitive impairments in autism are associated with neural processing deficits in specific brain regions. However, these studies have primarily focused on neural systems responsible for face processing and social behaviors. Although repetitive, stereotyped behaviors are a hallmark of autism, little is known about the neural mechanisms underlying these behaviors in the disorder. METHODS: We used functional magnetic resonance imaging (fMRI) to investigate the neural correlates of shifts in behavioral response and cognitive set in 18 individuals with high functioning autism and 15 neurotypical control participants. Participants performed a target detection task specifically designed to distinguish shifts in response from shifts in cognitive set. RESULTS: Individuals with autism showed lower accuracy on response shifting trials, independent of whether those trials also required a shift in cognitive set. Compared with control subjects, participants with autism showed reduced activation in frontal, striatal, and parietal regions during these trials. In addition, within the autism group, the severity of restricted, repetitive behaviors was negatively correlated with activation in anterior cingulate and posterior parietal regions. CONCLUSIONS: These results suggest that executive deficits and, by extension, repetitive behaviors associated with autism might reflect a core dysfunction within the brain's executive circuitry. PMID- 17916329 TI - Rostral anterior cingulate cortex volume correlates with depressed mood in normal healthy children. AB - BACKGROUND: The rostral anterior cingulate cortex (rACC) has been implicated as a structural neural correlate of familial major depressive disorder, raising the possibility that the structure of this region may act as a biologic marker of depression vulnerability. The aim of the current study was to determine whether children and adolescents with depressive symptoms have lower rACC volume relative to those without symptoms and examine how a positive family history of depression affects this relationship. METHODS: One hundred twelve normal healthy children (59 boys, 53 girls), age 7 to 17, without a current diagnosis or history of depression or other psychiatric illness, were recruited from the community. Mood symptoms were collected using the Pediatric Behavior Scale, a parent- and teacher reported questionnaire. Volumetric measures of the rACC were generated using structural magnetic resonance imaging (MRI). The relationship of depressive symptoms and rACC volume was examined. RESULTS: 1) The rACC volume was significantly lower in boys with subclinical depressive symptoms compared with boys with no depressive symptoms, particularly on the left side (14.6% reduction; F = 8.90, p = .005). 2) There was a negative correlation of rACC volume and depressive symptoms in boys, a finding that was more robust in subjects with a positive family history of depression. 3) In girls, there was not a significant association of depressive symptoms and rACC volume. CONCLUSIONS: These findings lend further support to the notion that rACC structure may act as a biologic marker of vulnerability or trait marker of depression. PMID- 17916331 TI - Anterior cingulotomy for major depression: clinical outcome and relationship to lesion characteristics. AB - BACKGROUND: Anterior cingulotomy (ACING) is a neurosurgical treatment for chronic refractory depression, pain, and obsessive-compulsive disorder. Anterior cingulotomy involves the placement of bilateral lesions in the anterior cingulate under stereotactic guidance. Although a long-established therapeutic intervention, the optimal location and volume of lesions are not known, but it is generally believed that efficacious lesions interrupt the fibers of the cingulum bundle. METHODS: Using T2-weighted magnetic resonance imaging, we tested the hypothesis that lesions placed more anteriorly would be associated with a better clinical response. We also tested a secondary hypothesis that a superior clinical response would be associated with larger lesion volumes. RESULTS: When assessed 12 months following surgery, a superior clinical response was associated with more anterior lesions but, unexpectedly, with smaller lesion volumes. Specifically, the best clinical response was associated with total (right plus left hemisphere) lesion volumes of 1000 to 2000 mm(3) centered at Montreal Neurological Institute (MNI) coordinates (+/- 9,19,30). CONCLUSIONS: There is considerable evidence from neuroimaging studies that more rostral areas within the anterior cingulate cortex are functionally and structurally abnormal in patients with major depressive disorder. Anteriorly placed ACING lesions would target and modify function within such regions. It should not be assumed that larger lesions are associated with a better response. These findings of relationships between lesion characteristics and clinical response argue against the suggestion that ACING represents a placebo treatment. PMID- 17916330 TI - Neural responses to monetary incentives in major depression. AB - BACKGROUND: Reduced responsiveness to positive incentives is a central feature of Major Depressive Disorder (MDD). In the present study, we compared neural correlates of monetary incentive processing in unmedicated depressed participants and never-depressed control subjects. METHODS: Fourteen currently depressed and 12 never-depressed participants underwent functional magnetic resonance imaging while participating in a monetary incentive delay task. During the task, participants were cued to anticipate and respond to a rapidly presented target to gain or avoid losing varying amounts of money. RESULTS: Depressed and never depressed participants did not differ in nucleus accumbens (NAcc) activation or in affective or behavioral responses during gain anticipation. Depressed participants did, however, exhibit increasing anterior cingulate activation during anticipation of increasing gains, whereas never-depressed participants showed increasing anterior cingulate activation during anticipation of increasing loss. Depressed participants also showed reduced discrimination of gain versus nongain outcomes. CONCLUSIONS: The present findings indicate that although unmedicated depressed individuals have the capacity to experience positive arousal and recruit NAcc activation during gain anticipation, they also exhibit increased anterior cingulate cortex activation, suggestive of increased conflict during anticipation of gains, in addition to showing reduced discrimination of gain versus nongain outcomes. PMID- 17916332 TI - Nonverbal delayed recognition in the relatives of schizophrenia patients with or without schizophrenia spectrum. AB - BACKGROUND: There is increased interest in the study of cognitive deficits as possible endophenotypic markers for schizophrenia. The main goal of this study was to determine how familiality and schizophrenia spectrum personality symptomatology are related to performance of auditory and visuospatial delayed recognition memory tasks. METHODS: The study sample consisted of 162 subjects divided into five groups. The groups included 39 patients with a DSM-IV diagnosis of schizophrenia or schizophreniform disorder; first-degree relatives of schizophrenia patients, 22 with and 31 without schizophrenia spectrum personality traits; and healthy control subjects with no family history of psychosis, 22 with and 48 without schizophrenia spectrum traits. Auditory and visuospatial delayed recognition memory performance was assessed. RESULTS: Significant differences were observed between patients and healthy control subjects in both auditory [F(1,79) = 7.358 p = .008] and visual [F(1,47) = 34.67, p < .001] delayed recognition tasks. When comparing the four non-patient groups, auditory and visuospatial discriminability decreased as a function of familiality of schizophrenia (p < .05). Deficits were more pronounced in relatives with schizophrenia spectrum traits [auditory d = .7114; visual d = 1.0199]. CONCLUSIONS: A biological relationship to schizophrenia increases the likelihood of impaired delayed recognition memory. Likewise, poorer performance is associated with schizophrenia spectrum phenotype only when combined with familiality. PMID- 17916333 TI - Altered emotional behaviors in the diabetes mellitus OLETF type 1 congenic rat. AB - GPR10 is a G-protein-coupled receptor expressed in thalamic and hypothalamic brain regions, including the reticular thalamic nucleus (RTN) and periventricular nucleus (Pev), and the endogenous ligand for this receptor, prolactin-releasing peptide (PrRP), has demonstrated regulatory effects on the stress response. We produced a congenic rat by introducing the Dmo1 allele from the OLETF rat which encodes the amino acid sequences of GPR10 with a truncated NH2-terminus, into the Brown-Norway background. Using receptor autoradiography, we determined a lack of specific [125I]PrRP binding in the RTN and Pev of these mutant rats compared to the control rats. Furthermore, intracerebroventricular injection of PrRP did not induce a significant increase of c-fos-like immunoreactivity in the paraventricular nucleus of the mutant rats compared to the control rats. The mutant rats also displayed a less anxious-like phenotype in three behavioral based models of anxiety-like behavior (open field, elevated plus maze and defensive withdrawal test). These data show the mutant congenic rat, of which GPR10 neither binds nor responds to PrRP, expresses less anxious-like phenotypes. On the basis of these observations, the GPR10 might be a novel target for the developing new drugs against anxiety and/or other stress-related diseases. PMID- 17916334 TI - Regional specificity and practice: dynamic changes in object and spatial working memory. AB - Working memory (WM) tasks engage a network of brain regions that includes primary, unimodal, and multimodal associative cortices. Little is known, however, about whether task practice influences these types of regions differently. In this experiment, we used event-related fMRI to examine practice-related activation changes in different region types over the course of a scanning session while participants performed a delayed-recognition task. The task contained separate WM processing stages (encoding, maintenance, retrieval) and different materials (object, spatial), which allowed us to investigate the influence of practice on different component processes. We observed significant monotonic decreases, and not increases, in fMRI signal primarily in unimodal and multimodal regions. These decreases occurred during WM encoding and retrieval, but not during maintenance. Finally, regions specific to the type of memoranda (e.g., spatial or object) showed a lesser degree of sensitivity to practice as compared to regions activated by both types of memoranda, suggesting that these regions may be specialized more for carrying out processing within a particular modality than for experience-related flexibility. Overall, these findings indicate that task practice does not have a uniform effect on stages of WM processing, the type of WM memoranda being processed or on different types of brain regions. Instead, regions engaged during WM encoding and retrieval may have greater capacity for functional plasticity than WM maintenance. Additionally, the degree of specialization within brain regions may determine processing efficiency. Unimodal and multimodal regions that participate in both object and spatial processing may be specialized for flexible experience-related change, while those supporting primary sensorimotor processing may operate at optimal efficiency and are less susceptible to practice. PMID- 17916335 TI - Evidence for a role of GABAA receptor in the acute restraint stress-induced enhancement of spatial memory. AB - Stress exerts complex effects on learning and memory; however, the understanding of the molecular mechanisms involved in stress effects on brain and behavior is rather limited. In this study, we investigated the regulation of the activation of MAPK (mitogen-activated protein kinase) cascades in the rat brain by GABAA receptor in a learning and memory task under acute restraint stress conditions. We found that the acute restraint stress improved the performance of the rats in the Morris water maze. Furthermore, the acute restraint stress significantly increased the phosphorylation of ERK and JNK in the hippocampus and prefrontal cortex (PFC), but not in the striatum. The increase paralleled the time course of the decrease of the level of GABAA receptor alpha1 subunit. The increase of P-ERK levels was inhibited by the agonist of GABAA receptor, muscimol, and further increased by the antagonist of the receptor, bicuculline. However, neither muscimol nor bicuculline affected the levels of P-JNK and P-p38. Finally, injection of muscimol partly reversed the acute restraint stress-induced enhancement of performance in the Morris water maze, and injection of bicuculline improved it. These results demonstrated that the changes in ERK phosphorylation in hippocampus and PFC were regulated by GABAA receptor in a learning and memory paradigm under acute restraint stress conditions. PMID- 17916336 TI - Activation of the serotonin 1A receptor alters the temporal characteristics of auditory responses in the inferior colliculus. AB - Serotonin, like other neuromodulators, acts on a range of receptor types, but its effects also depend on the functional characteristics of the neurons responding to receptor activation. In the inferior colliculus (IC), an auditory midbrain nucleus, activation of a common serotonin (5-HT) receptor type, the 5-HT 1A receptor, depresses auditory-evoked responses in many neurons. Whether these effects occur differentially in different types of neurons is unknown. In the current study, the effects of iontophoretic application of the 5-HT 1A agonist 8 OH-DPAT on auditory responses were compared with the characteristic frequencies (CFs), recording depths, and control first-spike latencies of the same group of IC neurons. The 8-OH-DPAT-evoked change in response significantly correlated with first-spike latency across the population, so that response depressions were more prevalent in longer-latency neurons. The 8-OH-DPAT-evoked change in response did not correlate with CF or with recording depth. 8-OH-DPAT also altered the temporal characteristics of spike trains in a subset of neurons that fired multiple spikes in response to brief stimuli. For these neurons, activation of the 5-HT 1A receptor suppressed lagging spikes proportionally more than initial spikes. These results suggest that the 5-HT 1A receptor, by affecting the timing of the responses of both individual neurons and the neuron population, shifts the temporal profile of evoked activity within the IC. PMID- 17916337 TI - Amyloid beta deposition is related to decreased glucose transporter-1 levels and hippocampal atrophy in brains of aged APP/PS1 mice. AB - The amount of the glucose transporter type-1 (GLUT-1) is decreased in the hippocampus and cerebral cortex of AD patients. In this study we therefore wanted to investigate the causal relationship between beta-amyloid (Abeta), GLUT-1 and hippocampal atrophy in the brains of young (8 months) and old (18 months) APP/PS1 mice. METHODS: Abeta and GLUT-1 were visualized immunohistochemically. Abeta load, GLUT-1 amount, capillary density and GLUT-1 amount per capillary density were determined in cortical and hippocampal areas using computer-assisted analysis systems. Hippocampal atrophy was determined by calculating the width of the outer molecular layer of the dentate gyrus (DG). RESULTS: In 18-month-old APP/PS1 mice we found a reduced GLUT-1 amount in the hippocampus but no differences in capillary density. The DG of these mice contained the highest level of Abeta in combination with hippocampal atrophy, and a reduced GLUT-1 amount per capillary density. At 8 months, no differences were observed. The highest Abeta deposition was found in the DG, although fourfold less compared to 18-month-old mice. CONCLUSIONS: We conclude that the GLUT-1 amount and capillary density in both wild type and transgenic mice decrease due to ageing. Further, a decreased amount of GLUT-1 is caused by decreased GLUT-1 amount/capillary density and not due to a reduced capillary density. We suggest that Abeta load in the hippocampus precedes the reduction of GLUT-1. A certain level of Abeta must be reached in the hippocampus, before it affects GLUT-1 amount/capillary density leading to further impairment of energy metabolism and hippocampal atrophy. PMID- 17916339 TI - Pathway and gene ontology based analysis of gene expression in a rat model of cerebral ischemic tolerance. AB - Ischemic tolerance is a phenomenon whereby a sublethal ischemic insult [ischemic preconditioning (IPC)] provides robust protection against subsequent lethal ischemia. Activation of N-methyl-D-aspartate (NMDA) receptors and subsequent new gene transcription are required for tolerance. We utilized the NMDA antagonist, MK801, prior to the IPC stimulus to separate candidate genes from epiphenomenona. Rats were divided into four groups: vehicle/IPC (preconditioned), MK801/IPC (attenuated preconditioning), vehicle/sham (non-preconditioned), and MK801/sham (non-preconditioned). Hippocampi (5/group/time point) were harvested immediately after ischemia as well as 1, 4, and 24 h post-ischemia to profile gene expression patterns using microarray analyses. Extracted mRNAs were pooled and subsequently hybridized to Affymetrix arrays. In addition, groups of rats were sacrificed for Western blot analysis and histological studies. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and gene ontology (GO) analyses were used to identify functionally related groups of genes whose modulation was statistically significant, while hierarchical cluster analysis was used to visualize the fold expression within these groups. Significantly modulated pathways included: MAP kinase signaling pathway, Toll receptor pathway, TGF-beta signaling pathways, and pathways associated with ribosome function and oxidative phosphorylation. Our data suggest that the tolerant brain responds to subsequent ischemic stress by partially downregulating inflammatory and upregulating protein synthesis and energy metabolism pathways. PMID- 17916338 TI - fMRI evidence for both generalized and specialized components of attentional control. AB - A central question in the study of selective attention is whether top-down attentional control mechanisms are generalized or specialized for the type of information that is to be attended. The current study examined this question using a voluntary orienting task that cued observers to attend to either one of two locations or to one of two colors. Location (spatial) and color (nonspatial) conditions were presented either randomly intermixed within the same block of trials or in separate blocks. Functional magnetic resonance imaging revealed that directing attention to a location or to a color activated a network of overlapping dorsal frontal and parietal areas, previously implicated in attentional control. The pattern of observed overlap was not affected by the intermixed versus blocked presentation of location and color conditions. Although portions of the frontal-parietal network were more active in response to location cues than to color cues, a secondary analysis also revealed that medial dorsal frontal and parietal cortex were specifically engaged in shifting visual attention regardless of the cued dimension (location or color). Together, the present results support the conclusion that attentional control is the combination of a generalized network that works in concert with subregions of the frontoparietal network that are highly specialized for directing attention based on the content of the to-be-attended information. PMID- 17916340 TI - Explicit versus implicit gaze processing assessed by ERPs. AB - Gaze processing was investigated using event-related potentials in two different tasks in which front-view and 3/4-view faces were presented, with eyes gazing straight ahead or averted. Task alternated between an explicit gaze direction judgment and a judgment on head orientation where gaze was irrelevant. Accuracy and reaction times were affected by the congruency of gaze and head directions in both tasks suggesting gaze was processed implicitly in the head orientation task. In both tasks, larger P1 and N170 were found for 3/4-view faces compared to front view faces that were not due to the luminance or contrast of the pictures. The N170 was also larger for averted than straight gaze but for front-view faces only. In contrast, larger amplitudes for straight than averted gaze were reliably measured around 400-600 ms regardless of head orientation or task demands, and likely reflected the outcome of gaze processing. The results suggest that head orientation and gaze direction discrimination occur regardless of task demands and interact at the decision making level. Neural markers of head orientation occur before those for gaze direction and the early structural encoding stages of face processing are view-dependant. PMID- 17916341 TI - Rare isoform of alkaline phosphatase in necrotic granulomatous inflammation of liver. PMID- 17916342 TI - Effects of two different types of exercise on GH/IGF axis in athletes. Is the free/total IGF-I ratio a new investigative approach? AB - BACKGROUND: Human growth hormone (hGH) responds to bouts of exercise by increasing, while the insulin-like growth factor-I (IGF-I) responses are conflicting. METHODS: Twenty well-trained male cyclists completed a brief duration exercise (A: warm up+increasing workload until exhaustion, lasting 25 min) and a medium duration exercise (B: warm up+70-80%VO(2 max)+increasing workload until exhaustion, lasting 40 min). The immunoreactivity of plasma hGH, the IGF-I in its total and free fraction were measured before and at the end of the exercise, and the free/total IGF-I ratio response to the two cycling exercise bouts was examined. RESULTS: Both A and B demonstrated increased hGH (from 77+/ 122 to 544+/-327 and 28+/-68 to 369+/-276 pmol/l respectively) and total IGF-I (from 67+/-10 to 70+/-10 and 55+/-14 to 61+/-15 nmol/l respectively). The free IGF-I was decreased only in A (from 0.38+/-0.16 to 0.32+/-0.14 nmol/l). Both A and B demonstrated a decreased free/total IGF-I ratio (from 0.57+/-0.30 to 0.46+/ 0.22 and 0.61+/-0.37 to 0.52+/-0.29). CONCLUSION: Brief and medium duration physical exercise influences the hGH, the total and free IGF-I concentrations. The free/total IGF-I ratio was also influenced and it might be related to the GH/IGF system. Its investigation might be a way of studying the training condition. PMID- 17916343 TI - Characterization of circulating DNA in healthy human plasma. AB - BACKGROUND: The importance of circulating DNA has been recognized since the detection of mutated oncogene products in cancer patients; however, there is little information about circulating DNA in normal human plasma. We characterized circulating DNA in normal human plasma to obtain basic information. METHODS: Circulating DNA was purified from plasma samples obtained from 10 healthy donors and examined. Purified DNA was cloned and their sequence determined and analyzed. The terminal structure was examined by a labeling method. RESULTS: The DNA levels in normal plasma samples were quite low (3.6-5.0 ng/ml). All 556 clones analyzed were independent, and obtained from various chromosomes and various regions of the gene. The mean values of their length and GC content were 176 bp and 53.7%, respectively. Their 5' and 3' ends were rich in C and G, respectively, and they presented as 5' protruding forms of double-stranded DNA in plasma. CONCLUSION: Circulating DNA in normal human plasma is derived from apoptotic cells but not from necrotic cells. Structural characteristics of the circulating DNA might be associated with their stability in plasma. PMID- 17916344 TI - Bone marrow transplant relapse with loss of an allele. AB - BACKGROUND: Detection of the extent of chimerism after transplantation is an important method for monitoring the engraftment of donor stem cells or diagnosing relapse. METHODS: The AmpFlSTR Identifiler amplification kit (Applied Biosystem, CA) was used to perform STR-PCR for the study of engraftment. RESULTS: At 6 months post-transplantation, the peripheral blood genotype shows a mixture of donor and recipient alleles, consistent with disease relapse. Interestingly, the D18S51 locus shows 2 donor alleles and only 1 recipient allele, the other recipient allele is missing. To further confirm chromosome loss, we used the D18S53 and D18S1129 in the ABI PRISM Linkage Mapping Sets. The D18S53 locus showed the recipient allele (179 bp) and shared allele (171 bp). Interestingly, the D18S1129 locus showed almost only 1 recipient allele (243 bp); the other recipient allele and shared allele (251 bp) is missing. CONCLUSION: The case illustrates that chromosome loss in tumor cells during the course of disease may cause corresponding loss of an STR locus. This circumstance is a potential source of error in the interpretation of engraftment analysis, especially if only one informative allele is used to monitor engraftment. PMID- 17916345 TI - The role of MAPK signaling in patterning and establishing axial symmetry in the gastropod Haliotis asinina. AB - Gastropods are members of the Spiralia, a diverse group of invertebrates that share a common early developmental program, which includes spiral cleavage and a larval trochophore stage. The spiral cleavage program results in the division of the embryo into four quadrants. Specification of the dorsal (D) quadrant is intimately linked with body plan organization and in equally cleaving gastropods occurs when one of the vegetal macromeres makes contact with overlying micromeres and receives an inductive signal that activates a MAPK signaling cascade. Following the induction of the 3D macromere, the embryo begins to gastrulate and assumes a bilateral cleavage pattern. Here we inhibit MAPK activation in 3D with U0126 and examine its effect on the formation and patterning of the trochophore, using a suite of territory-specific markers. The head (pretrochal) region appears to maintain quadri-radial symmetry in U0126-treated embryos, supporting a role for MAPK signaling in 3D in establishing dorsoventral polarity in this region. Posterior (posttrochal) structures - larval musculature, shell and foot--fail to develop in MAPK inhibited trochophores. Inhibition of 3D specification by an alternative method--monensin treatment--yields similar abnormal trochophores. However, genes that are normally expressed in the ectodermal structures (shell and foot) are detected in U0126- and monensin-perturbed larvae in patterns that suggest that this region has latent dorsoventral polarity that is manifested even in the absence of D quadrant specification. PMID- 17916346 TI - CAF-1 is essential for Drosophila development and involved in the maintenance of epigenetic memory. AB - DNA synthesis during S-phase and upon DNA repair is accompanied by chromatin assembly. The chromatin assembly factor CAF-1 has been biochemically well characterized to deposit histones onto newly synthesized DNA. To gain insights into the in vivo functions of CAF-1 in Drosophila, we generated null mutants of the largest subunit of dCAF-1, dCAF-1-p180. We show that, unlike CAF-1 mutant yeast, dCAF-1-p180 mutant flies are hemizygous lethal. Removal of maternal dCAF-1 p180 activity by germline clones blocks oogenesis. Tissue-specific deletion of dCAF-1-p180 in the eye primordia disrupts eye development. In addition, reduction of dCAF-1-p180 activity suppresses gene silencing at heterochromatin and antagonizes Polycomb-mediated cell fate determination. Furthermore, heterozygous dCAF-1-p180 mutant flies display an increased sensitivity to gamma-irradiation and a reduced efficiency in recombinational double strand break (DSB) repair. Our experiments also show that human hCAF-1-p150 can rescue the dCAF-1-p180 mutant flies, demonstrating a functional conservation of eukaryotic CAF-1 activities in vivo. Together, our results establish that dCAF-1-p180 is an essential gene for Drosophila development and further underscore the importance of dCAF-1 in regulating gene expression and DNA repair in vivo. PMID- 17916347 TI - The C. elegans M3 neuron guides the growth cone of its sister cell M2 via the Kruppel-like zinc finger protein MNM-2. AB - The invariant cell-cell interactions occurring during C. elegans development offer unique opportunities to discover how growing axons may receive guidance cues from neighboring cells. The mnm-2 mutant was isolated because of its defects in the axon trajectory of the bilateral M2 pharyngeal neurons in C. elegans. We found that mnm-2 enhances the effects of many growth cone guidance mutations on these axons, suggesting that it performs a novel function during axon guidance. We cloned mnm-2 and found that it encodes a protein with three C2H2 zinc finger domains related to the Kruppel-like Factor protein family. mnm-2 is expressed only transiently in the M2 neuron, but exhibits a sustained expression in its sister cell, the M3 neuron. Strikingly, the expression of mnm-2 is not sustained in the M3 cell of the mnm-2 mutant, indicating that this gene positively regulates itself in that cell. Electropharyngeograms also indicate that the M3 cell is functionally impaired in the mnm-2 mutant. We used an M3-specific promoter to show that the M2 axon defect can be rescued by expression of mnm-2 in its sister cell M3. The same promoter was used to express the pro-apoptotic gene egl-1 to kill the M3 cell, which resulted in an M2 axon guidance defect similar to that found in the mnm-2 mutant. Our results suggest an M2 axon guidance model in which the M3 cell provides an important signal to the growth cone of its sister M2 and that this signal and the proper differentiation of M3 both depend on mnm-2 expression. This mechanism of axon guidance regulation allows fine tuning of trajectories between sister cells. PMID- 17916348 TI - Smad signaling in the neural crest regulates cardiac outflow tract remodeling through cell autonomous and non-cell autonomous effects. AB - Neural crest cells (NCCs) are indispensable for the development of the cardiac outflow tract (OFT). Here, we show that mice lacking Smad4 in NCCs have persistent truncus arteriosus (PTA), severe OFT cushion hypoplasia, defective OFT elongation, and mispositioning of the OFT. Cardiac NCCs lacking Smad4 have increased apoptosis, apparently due to decreased Msx1/2 expression. This contributes to the reduction of NCCs in the OFT. Unexpectedly, mutants have MF20 expressing cardiomyocytes in the splanchnic mesoderm within the second heart field (SHF). This may result from abnormal differentiation or defective recruitment of differentiating SHF cells into OFT. Alterations in Bmp4, Sema3C, and PlexinA2 signals in the mutant OFT, SHF, and NCCs, disrupt the communications among different cell populations. Such disruptions can further affect the recruitment of NCCs into the OFT mesenchyme, causing severe OFT cushion hypoplasia and OFT septation failure. Furthermore, these NCCs have drastically reduced levels of Ids and MT1-MMP, affecting the positioning and remodeling of the OFT. Thus, Smad-signaling in cardiac NCCs has cell autonomous effects on their survival and non-cell autonomous effects on coordinating the movement of multiple cell lineages in the positioning and the remodeling of the OFT. PMID- 17916349 TI - A dynamic gradient of Wnt signaling controls initiation of neurogenesis in the mammalian cortex and cellular specification in the hippocampus. AB - Neurogenesis in the developing neocortex is a strictly regulated process of cell division and differentiation. Here we report that a gradual retreat of canonical Wnt signaling in the cortex from lateral-to-medial and anterior-to-posterior is a prerequisite of neurogenesis. Ectopic expression of a beta-catenin/LEF1 fusion protein maintains active canonical Wnt signaling in the developing cortex and delays the expression onset of the neurogenic factors Pax6, Ngn2 and Tbr2 and subsequent neurogenesis. Contrary to this, conditional ablation of beta-catenin accelerates expression of the same neurogenic genes. Furthermore, we show that a sustained canonical Wnt activity in the lateral cortex gives rise to cells with hippocampal characteristics in the cortical plate at the expense of the cortical fate, and to cells with dentate gyrus characteristics in the hippocampus. This suggests that the dose of canonical Wnt signaling determines cellular fate in the developing cortex and hippocampus, and that recession of Wnt signaling acts as a morphogenetic gradient regulating neurogenesis in the cortex. PMID- 17916350 TI - Inducible nitric oxide synthase and cyclooxygenase-2 participate in anti inflammatory and analgesic effects of the natural marine compound lemnalol from Formosan soft coral Lemnalia cervicorni. AB - Lemnalol (8-isopropyl-5-methyl-4-methylene-decahydro-1,5-cyclo-naphthalen-3-ol) is a natural compound isolated from the marine soft coral Lemnalia cervicorni. In the present study, the anti-inflammatory and anti-nociceptive properties of lemnalol were investigated in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and carrageenan-injected rats, respectively. Our results demonstrate that lemnalol significantly inhibited the expression of the pro-inflammatory proteins, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), in LPS stimulated RAW 264.7 cells. An in vivo inflammation model was induced by intraplantar injection of carrageenan into rat hind paws. An intramuscular injection of lemnalol (15 mg/kg) 10 min before carrageenan injection resulted in significant inhibition of carrageenan-induced rat paw edema and thermal hyperalgesia behavior. Western blot experiments revealed that the carrageenan induced expression of iNOS and COX-2 in paw tissue was significantly down regulated by lemnalol. Moreover, post-intrathecal injection of lemnalol produced a dose-dependent anti-nociceptive effect in carrageenan-injected rats (1 and 5 microg). The present results indicate that the marine-derived compound lemnalol had anti-inflammatory and analgesic effects in LPS-stimulated RAW 264.7 cells and carrageenan-injected rats, respectively. In addition, inhibition of elevated iNOS and COX-2 protein expression as well as neurophil infiltration of carrageenan injected paws may be involved in the beneficial effects of lemnalol. PMID- 17916351 TI - 24 bp duplication of CHIT1 gene is not correlated with coronary artery disease in Corsica Island (France). AB - In this study we analyzed allele and genotype distributions of 24 bp duplication of the CHIT1 gene in a sample of patients (N=300) with coronary artery disease (CAD) and in a control group (N=300) from central Corsica (France), with the aim to investigate the possible association between CHIT1 genotypes and CAD in Corsican population. Serum chitotriosidase activity is increased in individuals experiencing an ischemic stroke of atherothrombotic etiology and in subjects with ischemic heart disease. Our results suggest that 24 bp duplication of CHIT1 gene is not correlated with CAD in Corsican population, according to a previous study carried out on a Spanish sample. Gene prevalence and perhaps gene-disease associations vary according to ethnicity. Further studies, based on different ethnic groups, could be suitable to determine the implication of this polymorphism with respect to CAD. PMID- 17916352 TI - Id-1 promotes TGF-beta1-induced cell motility through HSP27 activation and disassembly of adherens junction in prostate epithelial cells. AB - Id-1 (inhibitor of differentiation or DNA binding-1) has been positively associated with cell proliferation, cell cycle progression, and invasiveness during tumorigenesis. In addition, Id-1 has been shown to modulate cellular sensitivity to TGF-beta1 (transforming growth factor beta1). Here we demonstrate a novel role of Id-1 in promoting TGF-beta1-induced cell motility in a non malignant prostate epithelial cell line, NPTX. We found that Id-1 promoted F actin stress fiber formation in response to TGF-beta1, which was associated with increased cell-substrate adhesion and cell migration in NPTX cells. In addition, this positive effect of Id-1 on TGF-beta1-induced cell motility was mediated through activation of MEK-ERK signaling pathway and subsequent phosphorylation of HSP27 (heat shock protein 27). Furthermore, Id-1 disrupted the adherens junction complex in TGF-beta1-treated cells through down-regulation of E-cadherin, redistribution of beta-catenin, along with up-regulation of N-cadherin. These lines of evidence reveal a novel tumorigenic role of Id-1 through reorganization of actin cytoskeleton and disassembly of cell-cell adhesion in response to TGF beta1 in human prostate epithelial cells, and suggest that intracellular Id-1 levels might be a determining factor for switching TGF-beta1 from a growth inhibitor to a tumor promoter during prostate carcinogenesis. PMID- 17916353 TI - Modulation of the cAMP signaling pathway after traumatic brain injury. AB - Traumatic brain injury (TBI) results in both focal and diffuse brain pathologies that are exacerbated by the inflammatory response and progress from hours to days after the initial injury. Using a clinically relevant model of TBI, the parasagittal fluid-percussion brain injury (FPI) model, we found injury-induced impairments in the cyclic AMP (cAMP) signaling pathway. Levels of cAMP were depressed in the ipsilateral parietal cortex and hippocampus, as well as activation of its downstream target, protein kinase A, from 15 min to 48 h after moderate FPI. To determine if preventing hydrolysis of cAMP by administration of a phosphodiesterase (PDE) IV inhibitor would improve outcome after TBI, we treated animals intraperitoneally with rolipram (0.3 or 3.0 mg/kg) 30 min prior to TBI, and then once per day for 3 days. Rolipram treatment restored cAMP to sham levels and significantly reduced cortical contusion volume and improved neuronal cell survival in the parietal cortex and CA3 region of the hippocampus. Traumatic axonal injury, characterized by beta-amyloid precursor protein deposits in the external capsule, was also significantly reduced in rolipram-treated animals. Furthermore, levels of the pro-inflammatory cytokines, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), were significantly decreased with rolipram treatment. These results demonstrate that the cAMP-PKA signaling cascade is downregulated after TBI, and that treatment with a PDE IV inhibitor improves histopathological outcome and decreases inflammation after TBI. PMID- 17916354 TI - Viral vectors for in vivo gene transfer in Parkinson's disease: properties and clinical grade production. AB - Because Parkinson's disease is a progressive degenerative disorder that is mainly confined to the basal ganglia, gene transfer to deliver therapeutic molecules is an attractive treatment avenue. The present review focuses on direct in vivo gene transfer vectors that have been developed to a degree that they have been successfully used in animal model of Parkinson's disease. Accordingly, the properties of recombinant adenovirus, recombinant adeno-associated virus, herpes simplex virus, and lentivirus are described and contrasted. In order for viral vectors to be developed into clinical grade reagents, they must be manufactured and tested to precise regulatory standards. Indeed, clinical lots of viral vectors can be produced in compliance with current Good Manufacturing Practices (cGMPs) regulations using industry accepted manufacturing methodologies, manufacturing controls, and quality systems. The viral vector properties themselves combined with physiological product formulations facilitate long-term storage and direct in vivo administration. PMID- 17916355 TI - Cyclic AMP regulates bicarbonate secretion in cholangiocytes through release of ATP into bile. AB - BACKGROUND & AIMS: Bicarbonate secretion is a primary function of cholangiocytes. Either adenosine 3',5'-cyclic monophosphate (cAMP) or cytosolic Ca(2+) can mediate bicarbonate secretion, but these are thought to act through separate pathways. We examined the role of the inositol 1,4,5-trisphosphate receptor (InsP3R) in mediating bicarbonate secretion because this is the only intracellular Ca(2+) release channel in cholangiocytes. METHODS: Intrahepatic bile duct units (IBDUs) were microdissected from rat liver then luminal pH was examined by confocal microscopy during IBDU microperfusion. Cyclic AMP was increased using forskolin or secretin, and Ca(2+) was increased using acetylcholine (ACh) or adenosine triphosphate (ATP). Apyrase was used to hydrolyze extracellular ATP, and suramin was used to block apical P2Y ATP receptors. In selected experiments, IBDUs were pretreated with short interfering RNA (siRNA) to silence expression of specific InsP3R isoforms. RESULTS: Both cAMP and Ca(2+) agonists increased luminal pH. The effect of ACh on luminal pH was reduced by siRNA for basolateral (types I and II) but not apical (type III) InsP3R isoforms. The effect of forskolin on luminal pH was reduced by a cystic fibrosis transmembrane conductance regulator (CFTR) inhibitor and by siRNA for the type III InsP3R. Luminal apyrase or suramin blocked the effects of forskolin but not ACh on luminal pH. CONCLUSIONS: Cyclic AMP-induced ductular bicarbonate secretion depends on an autocrine signaling pathway that involves CFTR, apical release of ATP, stimulation of apical nucleotide receptors, and then activation of apical, type III InsP3Rs. The primary role of CFTR in bile duct secretion may be to regulate secretion of ATP rather than to secrete chloride and/or bicarbonate. PMID- 17916356 TI - Viral and host factors induce macrophage activation and loss of toll-like receptor tolerance in chronic HCV infection. AB - BACKGROUND & AIMS: Persistent inflammation contributes to progression of liver damage in chronic HCV (cHCV) infection. Repeated exposure to toll-like receptor (TLR) ligands results in tolerance, a protective mechanism aimed at limiting inflammation. METHODS: Monocytes/macrophages were repeatedly stimulated via proinflammatory cytokine-inducing TLRs and evaluated for activation markers. RESULTS: Unlike monocytes of controls or patients with nonalcoholic steatohepatitis, the monocytes of cHCV patients were hyperresponsive and failed to show homo- or heterotolerance to TLR ligands, manifested by elevated tumor necrosis factor (TNF)-alpha production. Serum levels of interferon (IFN)-gamma, endotoxin (TLR4 ligand), and HCV core protein (TLR2 ligand) were elevated in cHCV patients suggesting potential mechanisms for in vivo monocyte preactivation. Treatment of normal monocytes with IFN-gamma resulted in loss of tolerance to lipopolysaccharide (LPS) or HCV core protein. Furthermore, we found increased levels of MyD88-IRAK1 complexes and nuclear factor (NF)-kappaB activity both in monocytes of cHCV patients and in normal monocytes that lost TLR tolerance after IFN-gamma + LPS pretreatment. In vitro differentiation of TLR non-tolerant cHCV monocytes into macrophages restored their capacity to exhibit TLR tolerance to LPS and HCV core protein, and this could be reversed by administration of IFN gamma. cHCV patients exhibited increased TNF-alpha in the circulation and in the liver. In cHCV livers, we found Kupffer cell/macrophage activation indicated by increased CD163 and CD33 expression. CONCLUSIONS: We identified that host-derived factors (IFN-gamma and endotoxin) and viral factors (HCV core protein) act in tandem to induce and maintain monocyte/macrophage activation, thus favoring persistent inflammation in patients with cHCV infection. PMID- 17916360 TI - The role of vocal self-stimulation in female responses to males: implications for state-reading. AB - As research neurobiologists, we pursue specific questions, and the answers rendered are also correspondingly specific. Our goal, however, is to understand an entire system or the whole organism. To that end, it is not only useful, but sometimes also necessary, that we periodically reappraise a body of specific data in light of current knowledge of the field at large. In this spirit, the present paper reviews my work on the neural and hormonal mechanisms underlying the reproductive system of ring doves and others' studies of songbirds. By integrating these fields I then advance the concept that inherent in the avian breeding system is the mechanism of "state-reading" (a term fashioned after "mindreading", which was coined by cognitive neuroscientists). State-reading helps to coordinate a sequence of endocrine and behavioral events to realize a desired objective, in this case, successful reproduction. PMID- 17916358 TI - Functional characteristics of the bovine hypothalamic-pituitary-adrenal axis vary with temperament. AB - Hypothalamic-pituitary-adrenal (HPA) axis function, in Brahman heifers of differing temperament, was evaluated using separate challenges with CRH and ACTH. Exit velocity (EV) measurement was used to classify heifer temperament as calm [C; consisted of 6 slowest heifers (EV=1.05+/-0.05 m/s)] or temperamental [T; 6 fastest heifers (EV=3.14+/-0.22 m/s)]. During the 6 h prior to CRH challenge, areas under the ACTH (P=0.025) and cortisol (P<0.001) curves were greater in the temperamental heifers. Baseline cortisol (P<0.001) but not ACTH (P=0.10) differed between temperament groups. Following CRH challenge, areas under the ACTH (P=0.057) and cortisol (P<0.01) response curves were greater in the calm animals. The same animals were subjected to an ACTH challenge 14 d following their utilization in the CRH stimulation experiment. Prior to ACTH challenge, baseline cortisol concentrations were higher (P<0.001) in the temperamental heifers (T=18+/-2.6, C=4.3+/-0.6 ng/mL). Following ACTH administration, area under the cortisol response curve was greater (P=0.07) in the calm heifers. After declining below baseline concentrations during the post-challenge recovery period, cortisol in temperamental animals was again greater (P=0.02) than in the calm heifers. These data demonstrate that cattle with an excitable temperament exhibit increased stress responsiveness to handling, increased baseline adrenal function but not increased basal pituitary function, and a muted responsiveness to pharmacological stimulus. Thus, functional characteristics of the HPA axis vary with animal temperament. PMID- 17916357 TI - Pelvic nerve input mediates descending modulation of homovisceral processing in the thoracolumbar spinal cord of the rat. AB - BACKGROUND & AIMS: Colonic afferents project to the lumbosacral and thoracolumbar spinal cord via the pelvic and hypogastric/lumbar colonic nerves, respectively. Both spinal regions process inflammatory colonic stimuli. The role of thoracolumbar segments in processing acute colorectal pain is questionable, however, because the lumbosacral spinal cord appears sufficient to process reflex responses to acute pain. Here, we show that activity in pelvic nerve colonic afferents actively modulates thoracolumbar dorsal horn neuron processing of the same colonic stimulus through a supraspinal loop: homovisceral descending modulation. METHODS: Dorsal horn neurons were recorded in the rat thoracolumbar spinal cord after acute or chronic pelvic neurectomy and cervical cold block. RESULTS: Acute pelvic neurectomy or lidocaine inhibition of lumbosacral dorsal roots facilitated the excitatory response of thoracolumbar dorsal horn neurons to colorectal distention (CRD) and decreased the percentage of neurons inhibited by CRD, suggesting colonic input over the pelvic nerve inhibits thoracolumbar processing of the same stimulus. Ectopic activity developed in the proximal pelvic nerve after chronic neurectomy reactivating the inhibitory circuit, inhibiting thoracolumbar neurons. Cervical cold block alleviated the inhibition in intact or chronic neurectomized rats. However, the facilitated response after acute pelvic neurectomy was inhibited by cervical cold block, exposing an underlying descending facilitation. Inhibiting pelvic nerve input after cervical cold block had minimal effect. CONCLUSIONS: These data demonstrate that input over the pelvic nerve modulates the response of thoracolumbar spinal neurons to CRD by a supraspinal loop and that increasing thoracolumbar processing increases visceral hyperalgesia. PMID- 17916361 TI - Degree of counterion binding on water in oil microemulsions. AB - An ion-exchange process has been used to prepare HOT from NaOT (sodium bis(2 ethylhexyl)sulfosuccinate), where the Na(+) counterion has been replaced by H(+). The acidity function, H(0), of the aqueous core of HOT-based microemulsions decreases with W from H(0) approximately 0.6 at W>20 to H(0)=-1.4 at W=2. On the basis of the H(0) acidity function of the aqueous core and the dependence of H(0) on acid concentration, we have been able to determine the degree of counterion binding (beta) in microemulsions with a value of beta approximately 0.92 which is practically independent of the water content of the system. PMID- 17916362 TI - Sirt1 modulates premature senescence-like phenotype in human endothelial cells. AB - Yeast Sir2 plays critical roles in gene silencing, stress resistance and longevity. Mammalian Sirt1 NAD(+)-dependent protein deacetylase, the closest homolog of Sir2, regulates cell cycle, cellular senescence, apoptosis and metabolism, by functional interactions with a number of biological molecules such as p53. To investigate a role of Sirt1 in endothelial dysfunction and premature senescence, we examined the effects of Sirt1 inhibition in human umbilical vein endothelial cells (HUVEC). Sirt1 inhibition by sirtinol, which is a 2-hydroxy-1 napthaldehyde derivative, or siRNA for Sirt1-induced premature senescence-like phenotype, as judged by increased senescence-associated beta-galactosidase (SA beta-gal) activity, sustained growth arrest and enlarged and flattened cell morphology at 10 days after the treatment. Sixty-four percent of sirtinol (60 mumol/L)-treated HUVEC was SA-beta-gal-positive, whereas only 17% of vehicle treated cells were positive. Sirt1 inhibition by sirtinol or Sirt1 siRNA increased PAI-1 expression and decreased both protein expression and activity of eNOS. Treatment with sirtinol or Sirt1 siRNA increased acetylation of p53, while p53 expression was unaltered. Impaired epidermal growth factor-induced activation of mitogen-activated protein kinases was associated with Sirt1 inhibition-induced senescence-like growth arrest. Conversely, overexpression of Sirt1 prevented hydrogen peroxide-induced SA-beta-gal activity, morphological changes and deranged expression of PAI-1 and eNOS. These results showed that Sirt1 inhibition increased p53 acetylation and induced premature senescence-like phenotype in parallel with increased PAI-1 and decreased eNOS expression. Our data suggest that Sirt1 may exert protective effects against endothelial dysfunction by preventing stress-induced premature senescence and deranged expression of PAI-1 and eNOS. PMID- 17916363 TI - Relationship between propeptide pH unfolding and inhibitory ability during ProDer p 1 activation mechanism. AB - The major allergen Der p 1 of the house dust mite Dermatophagoides pteronyssinus is a papain-like cysteine protease (CA1) produced as an inactive precursor and associated with allergic diseases. The propeptide of Der p 1 exhibits a specific fold that makes it unique in the CA1 propeptide family. In this study, we investigated the activation steps involved in the maturation of the recombinant protease Der p 1 expressed in Pichia pastoris and the interaction of the full length and truncated soluble propeptides with their parent enzyme in terms of activity inhibition and BIAcore interaction analysis. According to our results, the activation of protease Der p 1 is a multistep mechanism that is characterized by at least two intermediates. The propeptide strongly inhibits unglycosylated and glycosylated recombinant Der p 1 (K(D)=7 nM) at neutral pH. This inhibition is pH dependent. It decreases from pH 7 to pH 4 and can be related to conformational changes of the propeptide characterized by an increase of its flexibility and formation of a molten globule state. Our results indicate that activation of the zymogen at pH 4 is a compromise between activity preservation and propeptide unfolding. PMID- 17916364 TI - Allosteric communication in dihydrofolate reductase: signaling network and pathways for closed to occluded transition and back. AB - Escherichia coli dihydrofolate reductase (DHFR) catalyzes the reduction of dihydrofolate to tetrahydrofolate. During the catalytic cycle, DHFR undergoes conformational transitions between the closed (CS) and occluded (OS) states that, respectively, describe whether the active site is closed or occluded by the Met20 loop. The CS-->OS and the reverse transition may be viewed as allosteric transitions. Using a sequence-based approach, we identify a network of residues that represents the allostery wiring diagram. Many of the residues in the allostery wiring diagram, which are dispersed throughout the adenosine-binding domain as well as the loop domain, are not conserved. Several of the residues in the network have been previously shown by NMR experiments, mutational studies, and molecular dynamics simulations to be linked to equilibration conformational fluctuations of DHFR. To further probe the nature of events that occur during conformational fluctuations, we use a self-organized polymer model to monitor the kinetics of the CS-->OS and the reverse transitions. During the CS-->OS transition, coordinated changes in a number of residues in the loop domain enable the Met20 loop to slide along the alpha-helix in the adenosine-binding domain. Sliding is triggered by pulling of the Met20 loop by the betaG-betaH loop and the pushing action of the betaG-betaH loop. The residues that facilitate the Met20 loop motion are part of the network of residues that transmit allosteric signals during the CS-->OS transition. Replacement of M16 and G121, whose C(alpha) atoms are about 4.3 A in the CS, by a disulfide cross-link impedes that CS-->OS transition. The order of events in the OS-->CS transition is not the reverse of the forward transition. The contact Glu18-Ser49 in the OS persists until the sliding of the Met20 loop is nearly complete. The ensemble of structures in the transition state in both the allosteric transitions is heterogeneous. The most probable transition-state structure resembles the OS (CS) in the CS-->OS (OS- >CS) transition, which is in accord with the Hammond postulate. Structures resembling the OS (CS) are present as minor ( approximately 1-3%) components in equilibrated CS (OS) structures. PMID- 17916365 TI - Affinity maturation increases the stability and plasticity of the Fv domain of anti-protein antibodies. AB - The somatic mutations accumulated in variable and framework regions of antibodies produce structural changes that increase the affinity towards the antigen. This implies conformational and non covalent bonding changes at the paratope, as well as possible quaternary structure changes and rearrangements at the V(H)-V(L) interface. The consequences of the affinity maturation on the stability of the Fv domain were studied in a system composed of two closely related antibodies, F10.6.6 and D44.1, which recognize the same hen egg-white lysozyme (HEL) epitope. The mAb F10.6.6 has an affinity constant 700 times higher than D44.1, due to a higher surface complementarity to HEL. The structure of the free form of the Fab F10.6.6 presented here allows a comparative study of the conformational changes produced upon binding to antigen. By means of structural comparison, kinetics and thermodynamics of binding and stability studies on Fab and Fv fragments of both antibodies, we have determined that the affinity maturation process of anti protein antibodies affects the shape of the combining site and the secondary structure content of the variable domain, stabilizes the V(H)-V(L) interaction, and consequently produces an increase of the Fv domain stability, improving the binding to antigen. PMID- 17916366 TI - Attitude of outpatients with neuromuscular diseases in Japan to pain and use of analgesics. AB - The prevalence of pain and its impact on outpatients with neuromuscular disease, and their attitude towards the use of analgesics were studied. Seventy-eight outpatients at the university hospital, Tokyo, diagnosed with Parkinson's disease, spinocerebellar degeneration, amyotrophic lateral sclerosis, or multiple sclerosis were asked whether they had experienced pain in the preceding week. The Brief Pain Inventory, Japanese version was used to interview participants reporting pain, about its intensity and interference with activities, the way they dealt with it, attitudes to pain and use of analgesics, and desire for treatment. Forty-six participants experienced pain in the preceding week (59%). The mean pain intensity was 4.1 out of 10, and 20% of participants reported that the degree of interference with mobility was at least 6 out of 10. Most participants dealt with their pain without medication, by changing posture frequently or massage. Approximately 80% of participants regarded pain as something they should endure. Half of the participants wanted more information on methods for pain relief. Approximately 80% of participants were anxious about adverse reactions of analgesics. These findings suggest that medical staffs should provide appropriate information and educate their patients. PMID- 17916367 TI - Higher amount of MyHC IIX in a wrist flexor in tetraplegic compared to hemiplegic cerebral palsy. AB - Spastic cerebral palsy can be divided into diagnostic groups by the relative severity of the arm impairment. This study investigates if hemiplegic, tetraplegic or diplegic cerebral palsy (CP) results in different patterns of myosin heavy chain (MyHC) expression in the flexor carpi ulnaris muscle from 17 young patients with CP. Using enzyme-immunohistochemistry and gel electrophoresis techniques we found a higher percentage of fibers expressing fast MyHC IIx (52%) in tetraplegic CP compared to hemiplegic patients (32%), (p<0.05). Tetraplegic CP also resulted in a lower amount of fibers expressing slow MyHC I (18%) compared to hemiplegic CP (40%), (p<0.005). The proportion of muscle fibers containing fetal MyHC was higher in tetraplegic CP compared to other groups, (p<0.005). Taken together theses results indicate that tetraplegic CP is associated with a shift from slow to fast myosins and that regenerative events are more prominent in tetraplegic CP compared with milder brain damage. PMID- 17916368 TI - Cardiovascular and pulmonary effects of NOS inhibition in endotoxemic conscious rats subjected to swimming training. AB - Sepsis is characterized by systemic hypotension, hyporeactiveness to vasoconstrictors, impaired tissue perfusion, and multiple organ failure. During exercise training (ET), dynamic cardiovascular adjustments take place to maintain proper blood pressure and adjust blood supply to different vascular beds. The aim of this study was to investigate whether ET protects against the cardiovascular abnormalities induced by LPS, a model of experimental endotoxemia, and to evaluate the role of nitric oxide (NO) in pulmonary edema. Wistar rats were subjected to swimming training (up to 1 h/day, 5 days/week for 4 weeks) after which their femoral artery and vein were catheterized. LPS (5 mg/kg, i.v.), injected in control (C) and trained animals (ET), promoted 3 distinct phases in mean arterial pressure (MAP) and heart rate (HR). After ET the alterations in MAP were attenuated. The ET animals showed a lower pulmonary edema index (PEI) after LPS (C=0.65+/-0.01; ET=0.60+/-0.02), which was attenuated after treatment with aminoguanidine in both groups (C=0.53+/-0.02; ET=0.53+/-0.02, p<0.05). After l NAME, PEI was enhanced numerically in the C and was statistically higher in the ET group (C=0.73+/-0.05; ET=1.30+/-0.3, p<0.05). 7-nitroindazole did not promote any alteration in either group. The adaptations promoted by ET seem to be beneficial, counteracting the cardiovascular abnormalities and pulmonary edema seen in septicemia induced by LPS. The results suggest that iNOS aggravates and cNOS protects against this pulmonary edema. PMID- 17916369 TI - Metallothionein, zinc, and mercury levels in tissues of young rats exposed to zinc and subsequently to mercury. AB - Several studies have described mercury toxicity and the role of metallothioneins (MT) in the detoxification and regulation of metal homeostasis. However, little data exist on this topic during the specific post-natal developmental phase in young mammals. This developmental phase is particularly important since young animals are more sensitive to toxicants than adults. The objective of this work was to investigate whether MT participates in the mechanism of protection conferred by zinc pre-treatment on the toxic effects induced by mercury in neonate rats. Pups were exposed to ZnCl(2) (5 doses of 27 mg/kg/day, s.c.) and subsequently to HgCl(2) (5 doses of 5 mg/kg/day, s.c.); metal (Zn and Hg) and MT contents were analyzed in the liver, kidney, and blood. MT was induced in the liver and kidney of pups of both Zn-sal and Zn-Hg groups, although the greatest increase was in neonates exposed to Zn only. A direct relationship exists between MT and metals for both hepatic and renal tissues, which indicates that the increase in metal levels occurs in parallel to the increase in MT content. Although the heat-treated cytosolic fraction is rich in MT and metals, higher Zn and Hg contents were detected in the insoluble fraction of all tissues. These results suggest that MT is, at least in part, responsible for preventing Hg accumulation in the liver and blood and decreasing renal toxicity. PMID- 17916370 TI - [Orbital epidermoid cyst. Case report]. AB - Epidermoid tumors represent 1% of all primary intracranial tumors. Most of them occur intradurally in cerebellopontine angle and parasellar citerns. The intra orbital location accounts for 4 to 5% of all primary intra-orbital tumors. We report the case of a 23-year-old girl with progressive right proptosis who had developed 6 months earlier. The tumor was removed via an external orbitotomy approach. Histology confirmed the diagnosis. PMID- 17916371 TI - [Epithelioid hemangioendothelioma mimicking an occipital artery aneurysm]. AB - INTRODUCTION: Epithelioid hemangioendothelioma (EHE) is a unique tumor of soft tissues, primarily occurring in adults. It is characterized by the proliferation of epithelioid or histiocytoid endothelial cells. Its clinical course being close to that of hemangioma and conventional angiosarcoma, the term epithelioid hemangioendothelioma was suggested. CASE REPORT: The authors report the case of 29-year-old women complaining of a right occipital nodule of the scalp, diagnosed later as an epithelioid hemangioendothelioma located in the scalp and appearing like an aneurysm of the occipital artery on clinical examination and on radiological investigations. On first clinical examination this tumor mimicked a sebaceous cyst. However during history taking we noted headaches and palpation revealed a pulsatile mass with a thrill. An aneurysm of the occipital artery was suspected according to ultra-sound and MRI findings. The treatment was a wide and complete excision of the lesion. Finally it was the histological examination which confirmed the diagnosis of epithelioid hemangioendothelioma. There was no evidence of local recurrence at the twelve-month follow-up. DISCUSSION: This case is unusual on the following points: a rare occurrence of this kind of tumors, especially in the scalp region, arterial origin of the tumor, but also the fact that it mimicked an aneurysm of the occipital artery. Epithelioid hemangioendotheliomas belong to the epithelioid vascular tumor spectrum. They have in common the morphologic epithelioid aspect of endothelial tumor cells. Cutaneous involvement is rare and often associated with multi-systemic localizations. PMID- 17916372 TI - [Hybrid osteosynthesis in orthognathic surgery: 28 cases of Le Fort I osteotomy]. AB - INTRODUCTION: The use of resorbable materials in orthognathic surgery is presently the object of numerous clinical and experimental studies. Their main drawbacks are their lack of rigidity and induced chronic inflammatory reactions. The purpose of this study was to propose and to test an original system of hybrid osteosynthesis associating a single titanium plate with three resorbable plates in Le Fort I osteotomies in the absence of maxillary expansion, or two titanium plates with two resorbable plates in case of maxillary expansion. METHODS: Our retrospective study concerned 28 patients operated between 2002 and 2005. A clinical follow-up of over one year detected the specific complications of the hybrid system and a cephalometric study analyzed the secondary relapse 5 months after surgery. These results were compared to the literature. RESULTS: One case of mobility and a case of instability of the jaw were observed, along with 5 cases of local chronic inflammatory reaction. The radiological results found a secondary sub-clinical relapse. The removal of the titanium material under local anesthesia was performed without difficulties in ambulatory care, which avoided a new hospitalization for the patient and a new general anesthesia. DISCUSSION: The hybrid system allowed obtaining a postoperative stability of the jaw comparable to that obtained by the exclusive use of titanium plates with an acceptable morbidity. This method of osteosynthesis combined the advantages of the resorbable material (removal of titanium plates under local anesthesia) and of titanium material (stability of the procedure). However this method requires a good experience of working with resorbable material, a good follow-up, and cooperative patients. These encouraging results urge us to extend the indications to mandibular sagittal osteotomy and to bi-maxillary osteotomies. PMID- 17916373 TI - Function analysis of sequences in human APOBEC3G involved in Vif-mediated degradation. AB - Human APOBEC3G (hA3G) has been identified as an anti-HIV cellular factor. As a counter measure, the HIV-1 protein Vif causes the degradation of hA3G by binding to it and directing it to the cellular proteasome. In this work, we have used hA3G deletion mutants to map the region in hA3G required for its degradation by Vif to hA3G amino acids 105-245, the linker region between the two zinc coordination motifs. A small fragment of hA3G containing only amino acids 105-245 will undergo Vif-induced degradation. However, while amino acids 105-156 of hA3G are required for Vif interaction with hA3G, they are not themselves sufficient for hA3G degradation, a process that further requires amino acids 157-245. While expression of hA3G fragments 1-156 or 157-384 (but not 246-384) can dominantly inhibit the Vif-mediated degradation of full-length hA3G, only the N-terminal fragment inhibits the Vif/hA3G interaction. Inhibition of hA3G degradation by the C-terminal hA3G fragment 157-384 appears to be related to its ability to prevent the polyubiquitination of hA3G induced by Vif, a process that is required for Vif mediated proteosomal degradation of hA3G. Non-permissive cells stably expressing hA3G 1-156 or hA3G 157-384 are able to inhibit the replication of wild-type HIV 1, thereby verifying the inhibitory effect of these fragments upon Vif-mediated hA3G degradation and suggesting their potential in anti-HIV-1 therapy. PMID- 17916374 TI - Biological and genetic analysis of a bovine-like coronavirus isolated from water buffalo (Bubalus bubalis) calves. AB - We describe the isolation, biological and genetic characterization of a host range variant of bovine coronavirus (BCoV) detected in water buffalo (Bubalus bubalis). By conventional and real-time RT-PCR assays, the virus was demonstrated in the intestinal contents of two 20-day-old buffalo calves dead of a severe form of enteritis and in the feces of additional 17 buffalo calves with diarrhea. Virus isolation, hemagglutination and receptor-destroying enzyme activity showed that the buffalo coronavirus (BuCoV) is closely related to BCoV but possesses some different biological properties. Sequence and phylogenetic analyses of the 3' end (9.6 kb) of the BuCoV RNA revealed a genomic organization typical of group 2 coronaviruses. Moreover, the genetic distance between BuCoV and BCoV was proven to be the same or even higher than the distance between other ruminant coronaviruses and BCoV. In conclusion, our data support the existence of a host range variant of BCoV associated with enteritis in buffaloes. PMID- 17916375 TI - Age-related change in the retinoid X receptor beta gene expression in peripheral blood mononuclear cells of healthy volunteers: effect of 13-cis retinoic acid supplementation. AB - The regulation of cell growth and differentiation and also expression of a number of genes by retinoids are mediated by nuclear retinoid receptors (RARs and/or RXRs). In this study we investigated age-related alteration in both RAR and RXR receptor subtypes gene expression and tissue transglutaminase (tTG) activity before and after supplementation with 13-cis retinoic acid (13cRA) in human peripheral blood mononuclear cells (PBMCs). Healthy men (40) were divided in two groups according to their age (young group: 26.1+/-4.1 years and old group: 65.4+/-3.8 years). Each volunteer received 13cRA (Curacne), 0.5mg/(kgday)) during a period of 4 weeks. We have shown that RXRbeta expression was decreased significantly (p=0.0108) in PBMCs of elderly men when compared to that of young volunteers. Distribution of retinoic acid receptor subtype expression in PBMCs was found in the order: RXRbeta>RARgamma>RXRalpha>RARalpha. The tTG activity in PBMCs reflected a trend to be enhanced after 13-cis retinoic acid supplementation. In conclusion, we demonstrate a significant decrease in the expression of RXRbeta subtype of rexinoid receptors in PBMCs of healthy elderly men. Our data suggest that in healthy elderly men reduction of RXRbeta expression in PBMCs might be a common feature of physiological senescence. PMID- 17916376 TI - Comparative study of central corneal thickness measurement with slit-lamp optical coherence tomography and visante optical coherence tomography. AB - PURPOSE: To evaluate the repeatability and reproducibility of central corneal thickness (CCT) measurements obtained by 2 anterior segment optical coherence tomography (OCT) imaging systems and to examine their agreements with ultrasound pachymetry. DESIGN: Observational cross-sectional study. PARTICIPANTS: Fifty eyes from 50 healthy normal subjects were recruited. METHODS: In one randomly selected eye in each subject, CCT was measured by slit-lamp OCT (SLOCT), Visante OCT, and ultrasound pachymetry. For anterior segment OCT measurements, both automatic and manual CCTs were obtained. Twenty-five of the 50 subjects were invited for 2 more visits within a week to evaluate repeatability and reproducibility of CCT measurement. MAIN OUTCOME MEASURES: Central corneal thickness measurement obtained by the 3 methods and their agreements. Intrasession and intersession within-subject standard deviation (S(w)), precision (1.96xS(w)), coefficient of variation (CV(w)) (100xS(w)/overall mean), and intraclass correlation coefficient (ICC) were calculated to evaluate repeatability and reproducibility. RESULTS: Good repeatability and reproducibility were found for both automatic and manual CCT measurements obtained by SLOCT and Visante OCT. For intrasession repeatability, CV(w) and ICC values ranged between 0.9% and 1.2% and 0.96 and 0.98, respectively. For intersession reproducibility, the respective CV(w) and ICC values ranged between 1.2% and 1.4% and 0.94 and 0.96. Although no significant difference was found between automatic/manual SLOCT measurements and ultrasound pachymetry, automatic Visante OCT CCT (535.7+/-30.2 microm) was significantly less than CCT with ultrasound pachymetry (550.3+/-31.14 microm) (P<0.001). In contrast, manual Visante OCT measurement (558.8+/-32.8 microm) was slightly higher than ultrasound pachymetry (P<0.001). Nevertheless, SLOCT and Visante OCT measurement of CCT had 95% limits of agreement comparable to that of ultrasound pachymetry. The best agreement was observed in the manual SLOCT measurement (95% limits of agreement between -15.5 and 11.7 microm). CONCLUSIONS: Both SLOCT and Visante OCT automatic and manual CCT measurements were reliable and showed comparable agreement with ultrasound pachymetry. Although the 2 anterior segment OCT imaging systems have similar design and working principles, clinicians should be aware of the differences in CCT measurement between the 2 anterior segment OCTs. PMID- 17916377 TI - Comparison of gonioscopy and anterior segment ocular coherence tomography in detecting angle closure in different quadrants of the anterior chamber angle. AB - PURPOSE: To compare the performance of gonioscopy and anterior segment (AS) optical coherence tomography (OCT) in detecting angle closure in the different quadrants of the anterior chamber angle (ACA). DESIGN: Cross-sectional observational study. PARTICIPANTS: Five hundred two consecutive subjects more than 50 years of age with no previous ophthalmic problems recruited from a community clinic in Singapore. METHODS: All subjects underwent gonioscopy and AS OCT imaging in the dark. Using gonioscopy, the ACA was graded using the Scheie system by a single examiner masked to AS OCT findings. MAIN OUTCOME MEASURES: The ACA in a particular quadrant was classified as closed if the posterior trabecular meshwork could not be seen on gonioscopy. A closed ACA on AS OCT imaging was defined by the presence of any contact between the iris and angle wall anterior to the scleral spur. RESULTS: After excluding eyes with poor image quality, a total of 423 right eyes were included in the analysis. A closed angle in at least 1 quadrant was observed in 59% of the eyes by AS OCT and in 33% of the eyes by gonioscopy (P<0.001), with fair agreement between the two methods (kappa = 0.40). The frequency of closed angles by AS OCT and gonioscopy were 48% versus 29% superiorly, 43% versus 22% inferiorly, 18% versus 14% nasally, and 12% versus 20% temporally, respectively. Of the 119 of 1692 quadrants that were closed on gonioscopy but open on AS OCT, a steep iris profile was present in 61 (51%) of 119 quadrants on AS OCT, and of the 276 of 1692 quadrants that were open on gonioscopy but closed on AS OCT, 196 (71%) of 276 quadrants showed short iridoangle contact on AS OCT. CONCLUSIONS: The highest rates of closed angles on gonioscopy and AS OCT images were observed in the superior quadrant. Anterior segment OCT tended to detect more closed ACAs than gonioscopy, particularly in the superior and inferior quadrants. Variations in the iris profile and level of iridoangle contact also may explain some of the differences seen between gonioscopy and AS OCT. PMID- 17916378 TI - Incidence of endophthalmitis after 20- and 25-gauge vitrectomy. AB - PURPOSE: To assess the incidence rate of endophthalmitis after 25-gauge pars plana vitrectomy and to compare it with the endophthalmitis rate after 20-gauge pars plana vitrectomy. DESIGN: Retrospective, interventional, comparative cohort study. PARTICIPANTS: Eight thousand six hundred one consecutive pars plana vitrectomy surgery patients. METHODS: Surgeries performed at a single institution between January 1, 2004, and September 1, 2006, were reviewed. MAIN OUTCOME MEASURES: Incidence of postvitrectomy endophthalmitis. RESULTS: Endophthalmitis developed in 1 of 5498 eyes after 20-gauge vitrectomy (0.018%) and in 7 of 3103 eyes after 25-gauge vitrectomy cases (0.23%; P = 0.004). Median final visual acuity was counting fingers or hand movements (range, 20/50-no light perception), with comparable results between 20-gauge and 25-gauge endophthalmitis cases. CONCLUSIONS: The visual outcomes of vitrectomy-associated endophthalmitis, for both 20-gauge and 25-gauge vitrectomy, is poor. In this study population, 25 gauge vitrectomy had a statistically significant 12-fold higher incidence of endophthalmitis compared with 20-gauge vitrectomy. PMID- 17916379 TI - Beta-1 and beta-2 adrenoceptor polymorphisms: functional importance, impact on cardiovascular diseases and drug responses. AB - Beta-1 and beta-2 adrenoceptors (AR) play a pivotal role in regulation of the activity of the sympathetic nervous system and agonists and antagonists at both beta AR subtypes are frequently used in treatment of cardiovascular diseases. Both beta-1 and beta-2 AR genes have several polymorphisms that encode different amino acids. This review summarizes new insights into the functional importance of these polymorphisms, as well as their relationship to cardiovascular diseases and their impact on responses to adrenergic drug treatment. At present, it seems that, for cardiovascular diseases, beta-1 and beta-2 AR polymorphisms do not play a role as disease-causing genes; they might, however, be associated with disease related phenotypes. In addition they could influence adrenergic drug responses. Thus, the Arg389Gly beta-1 AR polymorphism might predict responsiveness to beta-1 AR agonist and blocker treatment: patients homozygous for the Arg389 beta-1 AR polymorphism should be good responders, while patients homozygous for the Gly389 beta-1 AR polymorphism should be poor or nonresponders. Furthermore, the Arg16Gln27 beta-2 AR seems to have strong impact on long-term agonist-induced beta-2 AR desensitization. Thus, patients carrying this haplotype appear to suffer from rapid loss of therapeutic efficacy of chronic agonist treatment, as has been demonstrated in asthma patients. Moreover, the Arg16Gln27 beta-2 AR haplotype might have some predictive value for poor outcome of heart failure. Future large prospective studies have to replicate these findings in order to reach the final goal of pharmacogenomic research: to optimize and individualize drug therapy based on the patient's genetic determinants of drug efficacy. PMID- 17916380 TI - Initiation of locomotion in lampreys. AB - The spinal circuitry underlying the generation of basic locomotor synergies has been described in substantial detail in lampreys and the cellular mechanisms have been identified. The initiation of locomotion, on the other hand, relies on supraspinal networks and the cellular mechanisms involved are only beginning to be understood. This review examines some of the findings relative to the neural mechanisms involved in the initiation of locomotion of lampreys. Locomotion can be elicited by sensory stimulation or by internal cues associated with fundamental needs of the animal such as food seeking, exploration, and mating. We have described mechanisms by which escape swimming is elicited in lampreys in response to mechanical skin stimulation. A rather simple neural connectivity is involved, including sensory and relay neurons, as well as the brainstem rhombencephalic reticulospinal cells, which act as command neurons. We have shown that reticulospinal cells have intrinsic membrane properties that allow them to transform a short duration sensory input into a long-lasting excitatory command that activates the spinal locomotor networks. These mechanisms constitute an important feature for the activation of escape swimming. Other sensory inputs can also elicit locomotion in lampreys. For instance, we have recently shown that olfactory signals evoke sustained depolarizations in reticulospinal neurons and chemical activation of the olfactory bulbs with local injections of glutamate induces fictive locomotion. The mechanisms by which internal cues initiate locomotion are less understood. Our research has focused on one particular locomotor center in the brainstem, the mesencephalic locomotor region (MLR). The MLR is believed to channel inputs from many brain regions to generate goal directed locomotion. It activates reticulospinal cells to elicit locomotor output in a graded fashion contrary to escape locomotor bouts, which are all-or-none. MLR inputs to reticulospinal cells use both glutamatergic and cholinergic transmission; nicotinic receptors on reticulospinal cells are involved. MLR excitatory inputs to reticulospinal cells in the middle (MRRN) are larger than those in the posterior rhombencephalic reticular nucleus (PRRN). Moreover at low stimulation strength, reticulospinal cells in the MRRN are activated first, whereas those in the PRRN require stronger stimulation strengths. The output from the MLR on one side activates reticulospinal neurons on both sides in a highly symmetrical fashion. This could account for the symmetrical bilateral locomotor output evoked during unilateral stimulation of the MLR in all animal species tested to date. Interestingly, muscarinic receptor activation reduces sensory inputs to reticulospinal neurons and, under natural conditions, the activation of MLR cholinergic neurons will likely reduce sensory inflow. Moreover, exposing the brainstem to muscarinic agonists generates sustained recurring depolarizations in reticulospinal neurons through pre-reticular effects. Cells in the caudal half of the rhombencephalon appear to be involved and we propose that the activation of these muscarinoceptive cells could provide additional excitation to reticulospinal cells when the MLR is activated under natural conditions. One important question relates to sources of inputs to the MLR. We found that substance P excites the MLR, whereas GABA inputs tonically maintain the MLR inhibited and removal of this inhibition initiates locomotion. Other locomotor centers exist such as a region in the ventral thalamus projecting directly to reticulospinal cells. This region, referred to as the diencephalic locomotor region, receives inputs from several areas in the forebrain and is likely important for goal-directed locomotion. In summary, this review focuses on the most recent findings relative to initiation of lamprey locomotion in response to sensory and internal cues in lampreys. PMID- 17916381 TI - Postnatal ontogeny of expression of the corticosteroid receptor genes in mammalian brains: inter-species and intra-species differences. AB - Corticosteroids are important mediators of homeostasis and stress, and exert their effects via two transcription-factor receptors, mineralocorticoid receptor (MR) and glucocorticoid receptor (GR). Both receptors are expressed in the brain in a region-specific manner, and regulate neuroendocrine and behavioral functions. Stress during early development has been demonstrated to lead to long term alterations in MR and GR levels and in the phenotypes that they mediate. To date, however, nearly all of this evidence has been obtained in rats, and there is actually no clear basis for extrapolation to other species. The current comparative review presents data, as available, on the following aspects of GR and MR gene expression in mouse and rat (Rodentia), tree shrew (Scandentia), common marmoset, squirrel monkey, rhesus macaque and human (Primates): (1) species-typical adult expression of MR mRNA and GR mRNA in hypothalamus, amygdala, hippocampus and neocortex; (2) species-typical neonate, infant, juvenile/adolescent and adult expression of MR mRNA and GR mRNA in hippocampus. (1) and (2) allow for identification of inter-species consistencies and differences in the relative levels of MR and GR expression across brain regions and ontogenetic stages. In addition, data are presented on (3) within-species inter-individual variation in MR and GR expression and causes thereof, including polymorphism and early life stress. Integrating the evidence in (1)-(3), it is noted that, should the expression levels of MR and GR at the time of early-life stress determine the latter's effects on the formers' long-term expression levels and functioning, then the long-term effects of early life stress on corticosteroid receptor expression and function will be species-, brain-region- and receptor-type-specific. PMID- 17916382 TI - Neural bases of goal-directed locomotion in vertebrates--an overview. AB - The different neural control systems involved in goal-directed vertebrate locomotion are reviewed. They include not only the central pattern generator networks in the spinal cord that generate the basic locomotor synergy and the brainstem command systems for locomotion but also the control systems for steering and control of body orientation (posture) and finally the neural structures responsible for determining which motor programs should be turned on in a given instant. The role of the basal ganglia is considered in this context. The review summarizes the available information from a general vertebrate perspective, but specific examples are often derived from the lamprey, which provides the most detailed information when considering cellular and network perspectives. PMID- 17916383 TI - Investigation of coenzyme Q biosynthesis in human fibroblast and HepG2 cells. AB - Coenzyme Q (CoQ) deficiency occurs in genetic disorders, during aging and various diseases. Diagnosis requires skin fibroblasts in tissue culture. [3H]Mevalonate incorporation was appropriate to measure the rate of CoQ synthesis in fibroblasts and hepatoblastoma cells. [14C]p-Hydroxybenzoate had limited permeability, but it could be increased with Fugene and cyclodextrin. Inhibition of decaprenyl-4 hydroxybenzoate transferase results in the accumulation of decaprenyl diphosphate, an indicator of enzyme deficiency. Also, analysis of the corresponding mRNAs in this case is useful. In vitro assays to measure trans prenyltransferase and decaprenyl-4-hydroxybenzoate transferase activities are not available. Neither measurement of methyltransferases is reliable in human cells. In vitro reconstruction of CoQ synthesis, in opposite to cholesterol synthesis, proved to be unsuccessful. Thus, the biochemical characterization of the CoQ biosynthetic system in human cells is restricted to a few reliable analytical procedures. PMID- 17916384 TI - Premeditated aggression is associated with serum cholesterol in abstinent drug and alcohol dependent men. AB - Relationships between aggressive subtypes and lipids were explored in 18 adult males undergoing treatment for substance dependence. A positive association was observed between a measure of premeditated aggression and total cholesterol. This was in contrast to an inverse association between lower cholesterol and higher impulsivity and anxiety. PMID- 17916385 TI - Response inhibition and impulsivity in schizophrenia. AB - Response inhibition in schizophrenia remains controversial, with behavioral correlates largely unknown. Inpatients with schizophrenia and controls completed a stop task and an impulsiveness questionnaire. Slower inhibitory processes were evident in schizophrenia, but there was no association with impulsivity. The nature of inhibition and impulsivity in schizophrenia is complex, and could reflect schizophrenia subgroups or disease states. PMID- 17916386 TI - Ultimatum bargaining behavior of people affected by schizophrenia. AB - Forty-nine people suffering from schizophrenia performed an interactive bargaining task involving small monetary rewards, known in classical game theory as the Ultimatum Game. In this task, the subject, in the role of the Proposer, has to offer his or her (anonymous) counterpart, the Responder, a share of a given sum of money. If the Responder accepts the offer, then the sum is split accordingly between the two. Otherwise, if he or she decides to reject the offer, both receive nothing. The patients' strategic behavior in both roles was compared with that of healthy and clinical controls. It was hypothesized that cognitive deficits characterizing schizophrenia, together with difficulties in social judgment, would impair the patients' bargaining ability. We found that in general schizophrenic patients did not fully exploit their strategic power as Proposers. In contrast, as Responders, schizophrenic patients acted not significantly different from controls. Further investigation is needed to establish the links between cognitive and symptomatic mediators and strategic decision-making ability. PMID- 17916387 TI - Complicated grief and uncomplicated grief are distinguishable constructs. AB - With data from 242 mourners who received help post-loss and were at least 6 months removed from their loss, it was found that symptoms proposed as denoting complicated grief (CG) are distinguishable from reactions representing uncomplicated grief (UG). With data from a subgroup of 130 mourners, CG but not UG was found to relate to concurrent distress and disability. PMID- 17916389 TI - Amylin causes anorexigenic effects via the hypothalamus and brain stem in chicks. AB - This study was conducted to determine the effects of amylin on appetite-related processes in chicks. Broiler chicks were centrally and peripherally injected with amylin, and feed and water intake were quantified. Feed intake was reduced after both central and peripheral amylin, but water intake was not affected. To determine if the hypothalamus and brainstem were involved in the anorexigenic effect, chicks were centrally and peripherally injected with amylin, and c-Fos immunoreactivity was quantified in the lateral hypothalamus (LH), ventromedial hypothalamus (VMH), area postrema (AP) and the nucleus of the solitary tract (NTS). Amylin decreased c-Fos immunoreactivity in the LH, did not affect the VMH, and increased c-Fos immunoreactivity in the AP and NTS. To determine if alimentary transit time was affected, chicks received central amylin and were gavaged with chicken feed slurry containing a visible marker. Amylin-treated chicks had increased alimentary canal transit time. Chicks also responded to central amylin with increased anxiety-related behaviors and increased plasma corticosterone concentration. These results demonstrate that amylin affects feeding, alimentary canal transit, and behavior through hypothalamic and brainstem mechanisms in chicks. PMID- 17916390 TI - Morphological interaction between galanin-like peptide- and dopamine-containing neurons in the rat arcuate nucleus. AB - Galanin-like peptide (GALP) is a 60-amino acid neuropeptide that plays an important role in the neuronal regulation of feeding, energy balance and reproduction. GALP is produced in the hypothalamic arcuate nucleus, an area containing, amongst other neuron types, two populations of neurons in which we were interested: a population of GALP-containing neurons which regulate energy balance and reproduction, and a second population consisting of tuberoinfundibular dopaminergic neurons which suppress prolactin secretion from the adenohypophysis. To characterize morphologically the relationship between GALP and dopamine-containing neurons in the arcuate nucleus, a double immunofluorescence study was performed on cryosections from rat brain. Immunohistochemical double labeling studies revealed that GALP-immunoreactive nerve fibers made direct contact on tyrosine hydroxylase (TH)-containing neuronal cell bodies in the arcuate nucleus. These results suggest that GALP-containing neurons innervate tuberoinfundibular dopaminergic neurons. PMID- 17916388 TI - The dynamics of effector T cells and Foxp3+ regulatory T cells in the promotion and regulation of autoimmune encephalomyelitis. AB - The Th1/Th2 paradigm of T helper cell subsets had to be revised when IL-17 producing T cells (Th17) were identified as a distinct T helper cell lineage. Th17 cells are very efficient inducers of tissue inflammation and crucial initiators of organ-specific autoimmunity. Whereas Th17 cells promote autoimmune tissue inflammation, Foxp3+ regulatory T cells (T-reg) are necessary and sufficient to prevent autoimmunity throughout the life span of an individual. Here, we review recent findings of how responses of effector T cells and T-reg cells with a defined antigen-specificity develop in autoimmune encephalomyelitis. Moreover, Th17 cells and Foxp3+ T-reg seem to be dichotomously related in that TGF-beta induces Foxp3 in naive T cells, but TGF-beta and IL-6 together drive the generation of Th17 cells. Thus, we give an overview of how Th17 cells, induced Foxp3+ T-reg, as well as how naturally occurring T-reg cells might cooperate to promote and regulate autoimmune inflammation of the central nervous system (CNS). The monitoring of the population dynamics of these T cell subsets in reporter mice in vivo will enable us to revisit the pathogenic concept of autoimmune inflammation in the CNS and design rational and phase-specific therapeutic interventions. PMID- 17916391 TI - TF2 binds to the regulatory promoter of alkaline phosphatase in Dicytostelium. AB - Alkaline phosphatase (ALP) activity becomes restricted to PstO cells at the prestalk-prespore boundary during the later stages of development, suggesting a novel function in the regulation of prestalk cell differentiation. To identify regulatory control sequences within the alp promoter, a series of 5' and internal deletions were generated and fused to the LacZ reporter gene. In vitro assays of reporter activity from Dicytostelium transformants containing the deleted promoter-LacZ fusion constructs showed that the -683 to -468 bp sequence is required for proper activation of the reporter in developing slugs. To identify DNA-protein interactions involved in the regulation of alp, EMSAs were preformed using a series of short overlapping PCR probes that span the regulatory promoter sequence. A sequence-specific DNA-binding protein was identified that interacts with the -665 to -635 bp sequence. This DNA-binding protein was sequentially purified using DEAE-Sephacel, heparin-Sepharose, DNA Affinity, and gel filtration chromatography. A polypeptide with a molecular weight of 28 kDa was identified on an SDS-PAGE. The purified protein was identified as TF2 by mass spectrometry. TF2 may, therefore, bind to the regulatory promoter of alp and function in the developmental control of PstO differentiation in Dicytostelium. PMID- 17916392 TI - Embryonic stem cell differentiation studied by FT-IR spectroscopy. AB - We propose, here, an FT-IR method to monitor the spontaneous differentiation of murine embryonic stem (ES) cells in their early development. Principal component analysis and subsequent linear discriminant analysis enabled us to segregate stem cell spectra into separate clusters - corresponding to different differentiation times - and to identify the most significant spectral changes during differentiation. Between days 4 to 7 of differentiation, these spectral changes in the protein amide I band (1700-1600 cm(-1)) and in the nucleic acid absorption region (1050-850 cm(-1)) indicated that mRNA translation was taking place and that specific proteins were produced, reflecting the appearance of a new phenotype. The DNA/RNA hybrid bands (954 cm(-1) and 899 cm(-1)) were also observed, suggesting that the transcriptional switch of the genome started at this stage of differentiation. As confirmed by cytochemical assays, the FT-IR approach presented here allows to detect at molecular level the biological events of ES cell differentiation as they take place and to monitor in a rapid way the temporal evolution of the ES cell culture. PMID- 17916393 TI - CacyBP/SIP interacts with tubulin in neuroblastoma NB2a cells and induces formation of globular tubulin assemblies. AB - CacyBP/SIP, originally identified as a S100A6 (calcyclin) target, was later shown to interact with some other members of the S100 family as well as with Siah-1 and Skp1 proteins. Recently, it has been shown that CacyBP/SIP is up-regulated during differentiation of cardiomyocytes. In this work we show that the level of CacyBP/SIP is higher in differentiated neuroblastoma NB2a cells than in undifferentiated ones and that in cells overexpressing CacyBP/SIP the level of GAP-43, a marker of differentiation, was increased. Since the process of differentiation is accompanied by an extensive rearrangement of microtubules, we examined whether CacyBP/SIP interacted with tubulin. By applying cross-linking experiments we found that these two proteins bind directly. The dissociation constant of the tubulin-CacyBP/SIP complex determined by the surface plasmon resonance technique is 1.57 x 10(-7 )M which suggests that the interaction is tight. The interaction and co-localization of CacyBP/SIP and tubulin was also demonstrated by co-immunoprecipitation, affinity chromatography and immunofluorescence methods. Light scattering measurements and electron microscopy studies revealed that CacyBP/SIP, but not its homologue, Sgt1, increased tubulin oligomerization. Altogether, our results suggest that CacyBP/SIP, via its interaction with tubulin, might contribute to the differentiation of neuroblastoma NB2a cells. PMID- 17916395 TI - The use of Bartonella henselae-specific age dependent IgG and IgM in diagnostic models to discriminate diseased from non-diseased in Cat Scratch Disease serology. AB - Cat Scratch Disease (CSD) is caused by Bartonella henselae infection and is a common cause of regional lymphadenopathy. The diagnosis of CSD largely depends on serology, but is hampered by both low sensitivity and specificity of the applied IgG and IgM assays. Using an in-house ELISA, we detected a significant age dependent increase in the IgG levels in the general population compared to CSD patients. With this knowledge, we developed diagnostic models to differentiate diseased from non-diseased persons. Evaluation of these models using samples from PCR-positive patients (n=155) and age-matched controls (n=244) showed an important increase in the assay performance if the combination of the IgG and IgM results were taken into account. If the specificity was set at 98% the sensitivity was only 45% and 32% for the IgM and IgG ELISA, respectively but increased to 59% when these results were combined. Also the use of age-dependent factors further improved the clinical relevance of the outcome raising the sensitivity to 64%. Although the sensitivity of the ELISA remains low we conclude that the use of models using the combination of both IgM and IgG test results and age-depending factors can be a useful diagnostic tool in the serodiagnosis of CSD. PMID- 17916396 TI - Nanofiber technology: designing the next generation of tissue engineering scaffolds. AB - Tissue engineering is an interdisciplinary field that has attempted to utilize a variety of processing methods with synthetic and natural polymers to fabricate scaffolds for the regeneration of tissues and organs. The study of structure function relationships in both normal and pathological tissues has been coupled with the development of biologically active substitutes or engineered materials. The fibrillar collagens, types I, II, and III, are the most abundant natural polymers in the body and are found throughout the interstitial spaces where they function to impart overall structural integrity and strength to tissues. The collagen structures, referred to as extracellular matrix (ECM), provide the cells with the appropriate biological environment for embryologic development, organogenesis, cell growth, and wound repair. In the native tissues, the structural ECM proteins range in diameter from 50 to 500 nm. In order to create scaffolds or ECM analogues, which are truly biomimicking at this scale, one must employ nanotechnology. Recent advances in nanotechnology have led to a variety of approaches for the development of engineered ECM analogues. To date, three processing techniques (self-assembly, phase separation, and electrospinning) have evolved to allow the fabrication of nanofibrous scaffolds. With these advances, the long-awaited and much anticipated construction of a truly "biomimicking" or "ideal" tissue engineered environment, or scaffold, for a variety of tissues is now highly feasible. This review will discuss the three primary technologies (with a focus on electrospinning) available to create tissue engineering scaffolds that are capable of mimicking native tissue, as well as explore the wide array of materials investigated for use in scaffolds. PMID- 17916397 TI - Growth factor binding to the pericellular matrix and its importance in tissue engineering. AB - In addition to its typical role as a scaffold, molecular filter, and cell modulator, the pericellular matrix can bind bioactive molecules and serve as a repository, while regulating their activation, synthesis, and degradation. This review focuses on interactions between bioactives, specifically growth factors and cytokines, with various components of the pericellular matrix. For example, biglycan and betaglycan, proteoglycans of the pericellular matrix, and decorin, a proteoglycan of the interstitial extracellular matrix, bind and regulate the activity and availability of transforming growth factor-beta. From evidence presented in this paper, it is obvious that the presence of growth factors in the pericellular matrix is integral to the spatiotemporal coordination of cellular activities to ensure proper tissue/organ formation during wound healing. It is believed by many researchers that the delivery of the right growth factors at the right time is instrumental to the orchestration of tissue regeneration. Thus, the interplay between the pericellular environment and bioactive molecules provides an underutilized knowledge base in the design and creation of tissue engineered constructs. PMID- 17916398 TI - Site-specific modification and PEGylation of pharmaceutical proteins mediated by transglutaminase. AB - Transglutaminase (TGase, E.C. 2.3.2.13) catalyzes acyl transfer reactions between the gamma-carboxamide groups of protein-bound glutamine (Gln) residues, which serve as acyl donors, and primary amines, resulting in the formation of new gamma amides of glutamic acid and ammonia. By using an amino-derivative of poly(ethylene glycol) (PEG-NH(2)) as substrate for the enzymatic reaction with TGase it is possible to covalently bind the PEG polymer to proteins of pharmaceutical interest. In our laboratory, we have conducted experiments aimed to modify proteins of known structure using TGase and, surprisingly, we were able to obtain site-specific modification or PEGylation of protein-bound Gln residue(s) in the protein substrates. For example, in apomyoglobin (apoMb, myoglobin devoid of heme) only Gln91 was modified and in human growth hormone only Gln40 and Gln141, despite these proteins having many more Gln residues. Moreover, we noticed that these proteins suffered highly selective limited proteolysis phenomena at the same chain regions being attacked by TGase. We have analysed also the results of other published experiments of TGase-mediated modification or PEGylation of several proteins in terms of protein structure and dynamics, among them alpha-lactalbumin and interleukin-2, as well as disordered proteins. A noteworthy correlation was observed between chain regions of high temperature factor (B-factor) determined crystallographically and sites of TGase attack and limited proteolysis, thus emphasizing the role of chain mobility or local unfolding in dictating site-specific enzymatic modification. We propose that enhanced chain flexibility favors limited enzymatic reactions on polypeptide substrates by TGases and proteases, as well as by other enzymes involved in a number of site-specific post-translational modifications of proteins, such as phosphorylation and glycosylation. Therefore, it is possible to predict the site(s) of TGase-mediated modification and PEGylation of a therapeutic protein on the basis of its structure and dynamics and, consequently, the likely effects of modifications on the functional properties of the protein. PMID- 17916400 TI - On-line social decision making and antisocial behavior: some essential but neglected issues. AB - The last quarter century has witnessed considerable progress in the scientific study of social information processing (SIP) and aggressive behavior in children. SIP research has shown that social decision making in youth is particularly predictive of antisocial behavior, especially as children enter and progress through adolescence. In furtherance of this research, more sophisticated, elaborate models of on-line social decision making have been developed, by which various domains of evaluative judgment are hypothesized to account for both responsive decision making and behavior, as well as self-initiated, instrumental functioning. However, discussions of these models have neglected a number of key issues. In particular, the roles of nonconscious cognitive factors, learning and development, impulsivity and behavioral disinhibition, emotion, and other internal and external factors (e.g., pharmacological influences and audience effects) have been largely absent from scholarly writings. In response, this article introduces discussion of these factors and reviews their possible roles in on-line social decision making and antisocial behavior in youth. PMID- 17916399 TI - Heparin-mimetic sulfated peptides with modulated affinities for heparin-binding peptides and growth factors. AB - Heterogeneity in the composition and in the polydispersity of heparin has motivated the development of homogeneous heparin mimics, and peptides of appropriate sequence and chemical function have therefore recently emerged as potential replacements for heparin in selected applications. Here, we report the assessment of the binding affinities of multiple sulfated peptides (SPs) for a set of heparin-binding peptides (HBPs) and for vascular endothelial growth factor isoform 165 (VEGF165); these binding partners have application in the selective immobilization of proteins and in hydrogel formation through non-covalent interactions. Sulfated peptides were produced via solid-phase methods, and their affinity for the HBPs and VEGF165 was assessed via affinity liquid chromatography (ALC), surface plasmon resonance (SPR), and in selected cases, isothermal titration calorimetry (ITC). The shortest peptide, SP(a), showed the highest affinity binding of HBPs and VEGF165 in both ALC and SPR measurements, with slight exceptions. Of the investigated HBPs, a peptide based on the heparin binding domain of human platelet factor 4 showed greatest binding affinities toward all of the SPs, consistent with its stronger binding to heparin. The affinity between SP(a) and PF4(ZIP) was indicated via SPR (K(D)=5.27 microM) and confirmed via ITC (K(D)=8.09 microM). The binding by SP(a) of both VEGF and HBPs suggests its use as a binding partner to multiple species, and the use of these interactions in assembly of materials. Given that the peptide sequences can be varied to control binding affinity and selectivity, opportunities are also suggested for the production of a wider array of matrices with selective binding and release properties useful for biomaterials applications. PMID- 17916401 TI - First trimester uterine Doppler and three-dimensional ultrasound placental volume calculation in predicting pre-eclampsia. AB - OBJECTIVE: To compare the efficacy of uterine artery Doppler velocimetry and three-dimensional ultrasound placental volume calculation alone or in combination in predicting at 11-14 weeks of gestation those pregnancies who will develop pre eclampsia. STUDY DESIGN: This was a prospective study of 348 nulliparous women scheduled for a routine prenatal ultrasound examination at 11-14 weeks. Color and pulsed wave Doppler was used to obtain uterine artery flow velocity waveforms transabdominally and the mean pulsatility index (PI) of the uterine arteries was calculated. The placental volume was measured by three-dimensional ultrasound using the virtual organ computer-aided analysis. Outcome variables considered were pre-eclampsia and pre-eclampsia requiring delivery <32 weeks. RESULTS: Pre eclampsia developed in 4.1% of the patients studied and in 1.7% a delivery before 32 weeks was required. Placental volume resulted significantly lower in pregnancies who will develop pre-eclampsia (t=4.636, p<0.003) and this was particularly evident in those pregnancies delivering <32 weeks (t=9.704, p<0.0002). No relationship was found between placental volume and mean uterine artery PI (r=-0.08, p=0.327). Uterine artery PI and placental volume showed similar sensitivities in predicting pre-eclampsia (50% vs. 56%) and pre-eclampsia with delivery <32 weeks (66.7% vs. 66.7%). The combination of uterine artery PI and placental volume gave better results when compared to the single use of one of these parameters (pre-eclampsia sensitivity 68.7%, pre-eclampsia requiring delivery <32 weeks 83.3%). CONCLUSIONS: The combination of abnormal uterine artery Doppler and low placental volume at 11-14 weeks achieves better results than does either test alone in the prediction of pre-eclampsia. PMID- 17916402 TI - Insertion of a magnesium(II)-octacarboranyl(hexylsulfanyl) porphyrazine into liposomes: a physico-chemical study. AB - The synthesis, characterization and liposome insertion of a novel magnesium(II) carboranyl-porphyrazine, i.e. [2,3,7,8,12,13,17,18-octakis-(1,2-dicarba-closo dodecaboranyl)-hexylthio-5,10,15,20-porphyrazine]magnesium(II) complex, MgHECSPz, are described. MgHECSPz was designed to improve the potentiality in multiple approach anticancer therapy. Liposomal formulations with different surface charge were prepared as delivering agents. The obtained loaded vectors were characterized by DLS, SAXS, SANS and zeta potential measurements in order to define the overall properties and structural details of loaded liposomes. PMID- 17916403 TI - High prevalence of pain in patients with cancer in a large population-based study in The Netherlands. AB - At present, no definite conclusions can be drawn about the real extent of the pain suffered by cancer patients. A population-based study was conducted to obtain reliable information about the prevalence and severity of pain in cancer patients (all phases) and about predictors of pain. A representative sample of cancer patients was recruited in the area from a cancer registry. Pain was assessed by the Brief Pain Inventory (BPI). Adequacy of pain treatment was assessed with the Pain Management Index (PMI). We found that 55% of the 1429 respondents had experienced pain past week; in 44% (n=351), the pain was moderate to severe (BPI score>or= 4). Total prevalence of pain/moderate to severe pain was present in 49%/41% in patients with curative treatment >or=6 months ago, 57%/43% in patients with current curative treatment or treatment <6 months ago, 56%/43% in patients with current palliative anti-cancer treatment and in 75%/70% in patients for whom treatment was no longer feasible. Positive predictors of the prevalence of pain were lower education level, more advanced disease and haematological (excluding (non)-Hodgkin lymphoma), gastro-intestinal, lung, or breast malignancies. According to the PMI, analgesic treatment was inadequate in 42% of the patients. Negative predictors of adequate treatment were current curative anti-cancer treatment and low education level. CONCLUSION: A substantial proportion of cancer patients does suffer from moderate to severe pain and does not receive adequate pain treatment. PMID- 17916404 TI - Interleukin-4 induces specific pp-GalNAc-T expression and alterations in mucin O glycosylation in colonic epithelial cells. AB - Mucus hypersecretion occurs as a consequence of the Th2 immune response in epithelia, yet it was not previously known whether the degree of O-glycosylation was modulated under such conditions. A colonic carcinoma cell line LS174T was used to assess the effect of interleukin (IL)-4 on the mRNA levels of eight pp GalNAc-Ts. A three- to four-fold increase in pp-GalNAc-T1, T4, and T7 levels was observed. Lysates of untreated or IL-4-treated cells were examined for their ability to transfer GalNAc residues onto a peptide corresponding to the tandem repeat portion of human MUC2. The number of incorporated GalNAc residues was greater after incubation with lysates of IL-4-treated cells than with lysates of untreated cells. Mucin-like large glycoproteins secreted by IL-4-treated cells had higher binding capacity to PNA and VVA-B(4) than those secreted by untreated cells. The results indicated that IL-4-treated LS174T cells are able to produce mucins with a higher degree of O-glycosylation than untreated counterparts. PMID- 17916405 TI - Isolation and characterization of a benzoylformate decarboxylase and a NAD+/NADP+ dependent benzaldehyde dehydrogenase involved in D-phenylglycine metabolism in Pseudomonas stutzeri ST-201. AB - Following induction with D-phenylglycine both d-phenylglycine aminotransferase activity and benzoylformate decarboxylase activity were observed in cultures of Pseudomonas stutzeri ST-201. Induction with benzoylformate, on the other hand, induced only benzoylformate decarboxylase activity. Purification of the benzoylformate decarboxylase, followed by N-terminal sequencing, enabled the design of probes for hybridization with P. stutzeri ST-201 genomic DNA libraries. Sequencing of two overlapping genomic DNA restriction fragments revealed two open reading frames which were denoted dpgB and dpgC. Sequence alignments suggested that the genes encoded a thiamin-diphosphate-dependent decarboxylase and an aldehyde dehydrogenase, respectively. Both genes were isolated and expressed in Escherichia coli. The dpgB gene product was confirmed as a benzoylformate decarboxylase while the dpgC gene product was characterized as a NAD+/NADP+ dependent benzaldehyde dehydrogenase. In keeping with their high sequence identities (both greater than 85%) the kinetic properties of the two enzymes were similar to those of the homologous enzymes in the mandelate pathway of Pseudomonas putida ATCC 12633. However, Pseudomonas stutzeri ST-201 was unable to grow on either isomer of mandelate, and sequencing indicated that the dpgB gene did not form part of an operon. Thus it appears that the two enzymes form part of a d-phenylglycine, rather than mandelate, degrading pathway. PMID- 17916406 TI - Isomer-specific retinoic acid biosynthesis in HeLa cells expressing recombinant class I aldehyde dehydrogenases. AB - Retinal dehydrogenase type 1 (RALDH1) catalyzes the oxidation of all-trans and 9 cis retinal to the respective retinoic acids (RAs), whereas another member of the aldehyde dehydrogenase family, the phenobarbital-induced aldehyde dehydrogenase (PB-ALDH), is very poorly active. We have previously generated chimeras between these two enzymes that displayed selectivity for retinal isomers in crude bacterial extracts. To examine whether the selectivity of the recombinant enzymes is retained in intact cells, we first assessed whether retinoid-isomerizing activity is present in cultured eukaryotic cells. Our results demonstrate that the only RA isomers detected in RALDH1-expressing or non-expressing cells corresponded to the same steric conformation as the supplied retinoids, indicating a lack of measurable 9-cis/all-trans retinoid-isomerizing activity. Finally, HeLa cells transfected with RALDH1 derivatives that were retinal isomer selective in vitro produced only the corresponding RA isomers, establishing these enzymes as useful tools to assess the respective roles of the two RA isomers in vivo. PMID- 17916407 TI - Effects of prenatal exposure to the CB-1 receptor agonist WIN 55212-2 or CO on the GABAergic neuronal systems of rat cerebellar cortex. AB - The aim of this study was to assess the effects of prenatal exposures to cannabinoids or carbon monoxide (CO) in an animal experimental model reproducing the environmental conditions in which a fetus develops whose mother, during pregnancy, ingests by smoking low doses of cannabinoids or CO. Particular attention was devoted to analyses of the long-term effects of the exposures at the level of the cerebellar cortex, where already during prenatal development the GABAergic neuronal systems may be modulated by both cannabinoids and CO. Three groups of rats were subjected to the following experimental conditions: exposure to cannabinoids by maternal treatment during pregnancy with the cannabinoid CB-1 receptor agonist WIN 55212-2 (WIN) (0.5 mg/kg/day, s.c.); exposure to CO by maternal exposure during pregnancy to CO (75 parts per million, by inhalation); and exposure to WIN+CO at the above doses and means of administration; a fourth group was used as control. The body weight of dams, length of pregnancy, litter size at birth, body weight and postnatal mortality of pups were monitored in order to evaluate possible effects of the exposures on reproduction and on prenatal and postnatal development. In the different groups, the long-term effects of the exposures were studied in adult rats (120-150 days) by light microscopy analyses of the structure of the cerebellar cortex and of the distribution in the cortex of markers of GABAergic neurons, such as GAD and GABA itself. Results. Exposures to WIN or CO did not affect reproduction or prenatal/postnatal development. Moreover, the exposed rats showed no structural alterations of the cerebellar cortex and displayed qualitative distribution patterns of GAD and GABA immunoreactivities similar to those of the controls. However, quantitative analyses indicated significant changes of both of these immunoreactivities: in comparison with the controls, they were significantly increased in WIN-exposed rats and reduced in CO-exposed rats, but not significantly different in WIN+CO-exposed rats. The changes were detected in the molecular and Purkinje neuron layers, but not in the granular layer. Prenatal exposures of rats to WIN or CO, at doses that do not affect reproduction, general processes of development and histomorphogenesis of the cerebellar cortex, cause significant changes of GAD and GABA immunoreactivities in some GABAergic neuronal systems of the adult rat cerebellar cortex, indicating selective up-regulation of GABA-mediated neurotransmission as a long-term consequence of chronic prenatal exposures to cannabinoids or CO. Because the changes consist of overexpression or, vice versa, underexpression of these immunoreactivities, functional alterations of opposite types in the GABAergic systems of the cerebellum following exposure to WIN or CO can be postulated, in agreement with the results of behavioral and clinical studies. No changes in immunoreactivities were detected after prenatal exposure to WIN and CO in association. PMID- 17916408 TI - Nitric oxide and GABA mediate bi-directional cardiovascular effects of orexin in the nucleus tractus solitarii of rats. AB - The present study investigated the cardiovascular effects of orexin (OX)-A and OX B in the nucleus tractus solitarii (NTS) and delineated the engagement of nitric oxide (NO) and GABA in OX-induced cardiovascular responses. In adult male Sprague Dawley rats maintained under propofol anesthesia, microinjection bilaterally into the NTS of OX-A or OX-B evoked bi-directional cardiovascular effects in a dose dependent manner. At a lower dose (5 pmol), OX-A or OX-B decreased systemic arterial pressure (SAP), heart rate (HR), and power density of the vasomotor components of SAP signals, our experimental index for sympathetic neurogenic vasomotor tone. At higher doses (>20 pmol), these two compounds elicited cardiovascular excitatory responses. These bi-directional cardiovascular effects of OX were abolished by co-injection of an OX(1) receptor antagonist, 1-(2 methylbenzoxazol-6-yl)-3-[1,5]naphthyridin-4-yl-urea hydrochloride (SB-334867, 0.75 nmol) or the OX(2) receptor antiserum (1:20). In addition, the vasodepressor effects of low dose (5 pmol) OX-A or OX-B in the NTS were attenuated by a nitric oxide synthase (NOS) inhibitor, N(G)-nitro-l-arginine methyl ester (l-NAME, 5 nmol), a neuronal nitric oxide synthase (nNOS) inhibitor, 7-nitroindazole (2.5 pmol) or the soluble guanylate cyclase (sGC) inhibitor, 1H-[1,2,4]oxadiazole[4,3 alpha]quinoxalin-1-one (250 pmol). The vasopressor effects of high dose (200 pmol) OX were reversed by co-administration with GABA(A) or GABA(B) receptor antagonist, bicuculline methiodine (10 pmol) or 2-hydroxy saclofen (100 pmol), or l-NAME (5 nmol). Our results indicate that OX-A or OX-B elicited bi-directional cardiovascular effects via OX receptor-dependent mechanisms. The vasodepressor effects of OX were induced by the nNOS-derived NO and activation of sGC associated signaling pathway, whereas the vasopressor effects were mediated by interaction with GABAergic or nitrergic neurotransmission in the NTS. PMID- 17916409 TI - Microinjection of the preferential dopamine receptor D3 agonist 7-hydroxy-N,N-di n-propylaminotetralin hydrobromide into the hypothalamic medial preoptic area induced ejaculation in anesthetized rats. AB - The pivotal role of the medial preoptic area (MPOA) of the hypothalamus in the dopaminergic cerebral control of ejaculation has been investigated for years; nevertheless the function of different dopamine receptors subclasses and their exact interrelations merit additional research. One hundred nanograms of a preferential D(3) agonist 7-OH-DPAT (7-hydroxy-N,N-di-n-propylaminotetralin hydrobromide) was microinjected unilaterally into the MPOA of male rats anesthetized with urethane. An ejaculation-related response (bulbospongiosus muscles rhythmic contractions and/or seminal vesicle pressure increases and/or expulsion of a semen plug) was observed in 8 of 10 rats devoid of sexual stimuli, while a similar response was observed in only one rat administered with 10 ng of 7-OH-DPAT. The effect of 7-OH-DPAT 100 ng was mostly abolished by simultaneous MPOA microinjection of 300 ng of a preferential D(3) antagonist N-[(n-butyl-2 pyrrolidinyl) methyl]-1-methoxy-4-cyanonaphthalene-2-carboxamide tartrate (nafadotride). Our results support the hypothesis that supraspinal command of ejaculation is mediated by D(2)-like receptors, probably by D(3) receptors, in the MPOA, and draw attention to the idea of these receptors serving as a promising target for pharmacological treatment of human ejaculatory disorders. PMID- 17916410 TI - Sympathectomy decreases size and invasiveness of tongue cancer in rats. AB - The sympathetic nervous system plays a role in carcinogenesis wherein locally released sympathetic neurotransmitters affect proliferation, angiogenesis, vessel permeability, lymphocyte traffic and cytokine production. The present in vivo study was designed to investigate whether surgical sympathectomy, both unilateral and bilateral, had an effect on tumor growth, interstitial fluid pressure (IFP) and lymphatics in rat tongue cancer. We used 4-nitroquinoline-1-oxide (4-NQO) in drinking water for 19 weeks to induce tongue cancer in 20 Dark Agouti rats. After 11 weeks, one group underwent unilateral sympathectomy and another underwent bilateral sympathectomy, while the third group underwent sham surgery. By 19 weeks, tumors in the bilaterally sympathectomized (BL-SCGx) rats were significantly smaller (P<0.05), more diffuse in appearance and less invasive (P<0.05) compared with the large exophytic tumors in the sham-operated rats. The relative lymphatic area was significantly decreased (P<0.05) in tumors in the BL SCGx rats compared with the sham group. Interestingly, the tumors in rats that underwent unilateral or bilateral sympathectomy had a significantly lower (P<0.05) IFP than those in sham rats. Lack of tyrosine hydroxylase (TH) immunoreactive nerves and few neuropeptide Y (NPY) positive fibers indicate absence of sympathetic nerve fibers in the bilateral sympathectomized group. The peritumoral lymph vessel area was correlated with the tumor size (P<0.001), depth of invasion (P<0.001), weight of rats (P<0.005) and IFP (P<0.05). In conclusion, the present study presents evidence that deprivation of sympathetic nerves decreases tumor growth in rat tongue, probably caused by decreasing IFP and lymph vessel area. PMID- 17916411 TI - Homer splice variants modulation within cortico-subcortical regions by dopamine D2 antagonists, a partial agonist, and an indirect agonist: implication for glutamatergic postsynaptic density in antipsychotics action. AB - Homer proteins are linked to both dopamine and glutamate neurotransmission and are putatively involved in the mechanisms of action of psychoactive drugs. In the present study, we evaluated the effects of compounds differently impacting dopaminergic neurotransmission on the transcripts of different Homer isoforms in rat forebrain by means of in situ hybridization histochemistry. Animals were treated with the typical antipsychotic haloperidol 0.8 mg/kg, the atypical antipsychotic clozapine 15 mg/kg, the dopamine partial agonist aripiprazole 12 mg/kg and 30 mg/kg and the dopamine transporter inhibitor GBR12909 30 mg/kg in acute and chronic paradigms. Homer 1a and ania-3 were induced in the caudate putamen by acute administration of aripiprazole 12 mg/kg, while aripiprazole 30 mg/kg had no significant effects. Furthermore, acute haloperidol and GBR12909 induced both the splice variants of Homer 1 in the caudate-putamen. In the nucleus accumbens, aripiprazole 30 mg/kg and clozapine increased Homer 1a and ania-3 transcripts in the shell, whereas haloperidol induced expression of both isoforms in core and shell. Aripiprazole 30 mg/kg increased Homer 1a in the frontal cortex. Homer 1 splice variants were both up-regulated by GBR12909 in the frontal cortex, whereas GBR12909 induced only ania-3 in the parietal cortex. In the chronic paradigm, results showed a significant induction of Homer 1a and ania 3 in the striatum by haloperidol and aripiprazole. The constitutive Homer 2 isoform was overexpressed in the lateral septum by chronic administration of haloperidol and clozapine. In the cortex the expression of Homer 1a and ania-3 was down-regulated by chronic clozapine and aripiprazole. These results may indicate a differential modulation of Homer genes by compounds differently regulating dopaminergic neurotransmission in discrete regions of the rat forebrain and suggest that Homer could be a molecular marker of the involvement of the glutamatergic postsynaptic density in antipsychotic mechanisms of action. PMID- 17916414 TI - Diarylheptanoids from the rhizomes of Alpinia officinarum and their anticancer activity. AB - Two new diarylheptanoids (1, 2), together with two known analogs (3, 4), were isolated from the rhizomes of Alpinia officinarum. The new compounds were elucidated to be (5S)-5-hydroxy-7-(3, 4-dihydroxyphenyl)-1-phenyl-3-heptanone (1) and (5R)-5-hydroxy-7-(3-methoxy-4, 5-dihydroxyphenyl)-1-phenyl-3-heptanone (2) based on their spectral analysis. Compound 4 showed moderate cytotoxicity against human tumor cell lines, HepG2, MCF-7 and SF-268, while no significant effect were found for compounds 1-3. PMID- 17916413 TI - A 24-h access I.V. self-administration schedule of morphine reinforcement and the estimation of recidivism: Pharmacological modification by arecoline. AB - Central cholinergic neurons are known to play a role in the pharmacological actions of opiates. The purpose of the study was to determine whether the muscarinic receptor agonist arecoline, administered during morphine self administration, would mitigate the subsequent return to self-administration behavior. Rats self-administered increasing concentrations of morphine in operant chambers according to a schedule that permitted unlimited access to lever activated i.v. infusions on a continuous 24 h basis from 10 to 14 days. Abstinence was induced by discontinuation of the morphine solution and mild withdrawal symptoms were evident from 14 to 74 h. Thereafter the rats remained in their home cages for a 6-week period of protracted abstinence. They were then returned to the operant chambers where lever responding had no reward consequence. The cholinergic muscarinic agonist arecoline was administered twice daily (0.25 or 1 mg/kg, s.c.) throughout the self-administration schedule of morphine. Arecoline treatment partly decreased the self-administration of morphine, it prevented the abstinence-induced decrease in body weight, and it reduced lever responding after protracted withdrawal (by 56%). In animals already dependent on morphine, arecoline failed to alter ongoing self-administration behavior, but responding induced by lever reinstatement 6 weeks after withdrawal was significantly reduced (by 33%). There was a significant relationship between the degree of self-administration activity and the degree of lever responding during reinstatement after protracted abstinence. The results of this study support the role of cholinergic systems in self-administration behavior and context-induced post-withdrawal drug seeking. PMID- 17916412 TI - Synaptophysin and postsynaptic density protein 95 in the human prefrontal cortex from mid-gestation into early adulthood. AB - Previous studies of postnatal synaptic development in human frontal cortex have shown that synaptic density rises after birth, reaches a plateau in childhood and then decreases to adult levels by late adolescence. A similar pattern has been seen in nonhuman primate cortex. These earlier studies in human cortex are limited, however, by significant age gaps in study subjects at critical inflection points of the developmental curve. Additionally, it is unclear if synaptic development occurs in different patterns in different cortical layers in prefrontal cortex (PFC). The purpose of this study was to examine synaptic density in human PFC across development by measuring two synaptic marker proteins: synaptophysin (presynaptic), and postsynaptic density protein 95 (PSD 95; postsynaptic). Western blotting was used to assess the relative levels of synaptophysin and PSD-95 in dorsolateral PFC of 42 subjects, distributed in age from 18 weeks gestation to 25 years. In addition, synaptophysin immunoreactivity was examined in each layer of areas 9 and 46 of PFC in 24 subjects, ranging in age from 0.1-25 years. Synaptophysin levels slowly increased from birth until age 5 and then increased more rapidly to peak in late childhood around age 10. Synaptophysin subsequently decreased until the adult level was reached by mid adolescence, around age 16. PSD-95 levels increased postnatally to reach a stable plateau by early childhood with a slight reduction in late adolescence and early adulthood. The pattern of synaptophysin immunoreactivity seen with immunohistochemistry was similar to the Western experiments but the changes across age were more subtle, with little change by layer within and across age. The developmental patterns exhibited by these synaptic marker proteins expand upon previous studies of developmental synaptic changes in human frontal cortex; synaptic density increases steadily from birth to late childhood, then decreases in early adolescence to reach adult levels by late adolescence. PMID- 17916415 TI - [Staphylococcus aureus infections carrying Panton-Valentine leukocidin genes in hospitals: which statute should be given to these strains?]. AB - OBJECTIVES: The aim of the present study was to investigate the prevalence Staphylococcus aureus infections carrying Panton-Valentine leukocidin (LPV) genes in our hospital by screening patients that are hospitalised or admitted for consultation, as well as to study the characteristics of these strains and the respectively infected patients. METHODS: A descriptive and retrospective study over the course of a 14 month period was conducted. The isolates of S. aureus were tested for antimicrobial resistance, in which detection of the virulence gene was performed by way of PCR, such as is the case for gene luk-PV which encoding the LPV. The genetic diversity of the strains carrying gene, luk-PV, was determined by way of pulsed-field gel electrophoresis and by the MLVA (Multiple Loci VNTR Analysis; VNTR, Variable Number of Tandem Repeats) method. RESULTS: 7.14% of the S. aureus isolates carried genes for LPV, which are primarily sourced from surgery, emergency, and outpatient consultation services. The nature of the reported infections is often surface, immediately collected, and more rarely deep. Genotyping revealed three principal clones that were gathering 55% of the strains, which in turn highlighted transmission to the nursing staff. COMMENTS: These strains of S. aureus LPV+ have the capacity for diffusion and pathogenicity, which leads to the need to take some specific measures at hospitals: the tracking of the LPV during repeat or deep infections with S. aureus, possibly the search for conveyance and individual measures for the eradication of the strain. Lastly, it is necessary to increase the nursing staff's awareness of the appropriate hygiene measures when they come into contact with these patients. PMID- 17916416 TI - [Evaluation of the in vitro activity of two betalactams on the oxidative metabolism of polymorphonuclear neutrophils]. AB - STUDY AIMS: The aim was to evaluate the in vitro effects of amoxicillin and its combination with clavulanic acid, two beta-lactams intravenously injected, on the oxidative metabolism of polymorphonuclear neutrophils. These cells play the major role in the "respiratory burst" as they produce superoxide anion to kill the infectious agent. An activation of this process by the injected antibiotics could enhance the bactericidal action or explain some of adverse effects. MATERIALS AND METHODS: Two models were used to estimate the O(2)(-) amounts produced in the presence of the antimicrobial agents. In the cellular model, O(2)(-) was generated by neutrophils artificially stimulated or not (separated by a gradient centrifugation through Histopaque 1077). In the acellular model, O(2)(-) was produced by the xanthine-xanthine oxidase system. O(2)(-) was measured by spectrophotometry using the ferricytochrome C reduction. RESULTS: The O(2)(-) production by polymorphonuclear neutrophils was increased when both antibiotics were added to the reaction mixture. A significant activation of the cell oxidative metabolism was observed with amoxicillin using various stimulating agents, that was higher without stimulation and lower when amoxicillin and clavulanic acid were associated. CONCLUSION: Amoxicillin could either activate polymorphonuclear neutrophils NADPH-oxidase or cause its activation by a membrane effect, or interfere with the zymosan activation way. It could then be supposed that this antimicrobial agent intensified the bactericidal effects. PMID- 17916417 TI - [Comparison of three selective chromogenic media for Methicillin-Resistant Staphylococcus aureus detection]. AB - PURPOSE: Rapid detection of Methicillin-Resistant Staphylococcus aureus (MRSA) is of major importance in hospital hygiene. In order to reduce the response time from screening laboratories, new selective chromogenic media have been developed and marketed by major microbiology companies. In this context, an evaluation of their performances was needed. MATERIALS AND METHODS: Media produced by Bio-Rad Laboratories (MRSASelect), Becton Dickinson (CHROMagar MRSA) and bioMerieux (chromID MRSA) were studied by 203screening samples, 110 of which were MRSA positive. Each Stahylococcus aureus was identified by catalase detection, Staphytect Plus Dryspot latex agglutination test and free coagulase detection, in addition to mannitol fermentation and Voges-Proskauer tests in the case of doubtful identification. Resistance was verified by checking the inhibition zone diameter of under 20 mm on 30 microg cefoxitin disks. RESULTS: Bio-Rad Laboratories, Becton Dickinson and bioMerieux media read at 24 or 24/48 hours have a respective sensitivity of 91, 71 and 85/92%. The specificity of these media is 99, 100 and 99/93%. CONCLUSION: These media proved to be new powerful and rapid tools used in screening for MRSA detection. MRSASelect is one of the most effective media showing high sensitivity and specificity and the easiest interpretation. chromID MRSA exhibits similar performance but needs more time to be as effective as Bio-Rad media while CHROMagar MRSA isn't enough efficient with its slightly lack of sensitivity to perform a reliable screening. PMID- 17916419 TI - Accuracy of commercial systems for identification of Burkholderia pseudomallei versus Burkholderia cepacia. AB - Infection caused by Burkholderia pseudomallei or Burkholderia cepacia may result in fatal outcome unless the causative agent is accurately identified in a short period, which is critical for treatment. We evaluated the reliability of the commonly used commercial systems, API 20NE, VITEK 2, and WalkAway 96, for comparative identification of B. pseudomallei versus B. cepacia clinical isolates. Based on biochemical and molecular tests as reference methods, API 20NE was probably the most reliable, with an accuracy of 87% and 93%, for the identification of B. pseudomallei and B. cepacia, respectively. The VITEK 2 and WalkAway 96 systems resulted in a number of misidentification and, thus, were less reliable. The performance of each system and identification guidelines for B. pseudomallei and B. cepacia are discussed. Our study emphasized that laboratories should carefully interpret the identification of B. pseudomallei and B. cepacia when using commercial systems. PMID- 17916420 TI - Pharmacokinetics of polymyxin B1 in patients with multidrug-resistant Gram negative bacterial infections. AB - Polymyxin B is increasingly used clinically for the treatment of multidrug resistant Gram-negative infections, despite very limited understanding of its disposition in humans. The disposition of intravenous polymyxin B1 in 9 adult patients was characterized. Random blood samples (specifically timed in relation to the dose administered) were obtained, and the serum concentrations of polymyxin B1 were assayed using a validated methodology by liquid chromatography mass spectroscopy. The serum concentration profiles of all the patients were analyzed by a population pharmacokinetic analysis using the nonparametric adaptive grid program. The mean volume of distribution and elimination half-life were found to be 47.2 L and 13.6 h, respectively. This is the 1st case series to date in which the pharmacokinetics of polymyxin B1 after intravenous administration are described. The results of the series in conjunction with pharmacodynamic and susceptibility surveillance studies could facilitate an approach to the design of optimal dosing regimens. PMID- 17916421 TI - A new polymerase chain reaction/restriction fragment length polymorphism protocol for Plasmodium vivax circumsporozoite protein genotype (VK210, VK247, and P. vivax-like) determination. AB - For the molecular diagnosis of Plasmodium vivax variants (VK210, VK247, and P. vivax-like) using DNA amplification procedures in the laboratory, the choice of rapid and inexpensive identification products of the 3 different genotypes is an important prerequisite. We report here the standardization of a new polymerase chain reaction/restriction fragment length polymorphism technique to identify the 3 described P. vivax circumsporozoite protein (CSP) variants using amplification of the central immunodominant region of the CSP gene of this protozoan. The simplicity, specificity, and sensitivity of the system described here is important to determine the prevalence and the distribution of infection with these P. vivax genotypes in endemic and nonendemic malaria areas, enabling a better understanding of their phylogeny. PMID- 17916422 TI - Toward a quantitative DNA-based definition of pneumococcal pneumonia: a comparison of Streptococcus pneumoniae target genes, with special reference to the Spn9802 fragment. AB - The current shift from phenotypically toward genotypically based microbial diagnosis is not unproblematic. A novel quantitative real-time polymerase chain reaction (PCR) assay based on the Spn9802 DNA fragment was therefore developed for detection of Streptococcus pneumoniae. Out of 44 bacterial species, only S. pneumoniae and Streptococcus pseudopneumoniae were positive in Spn9802 PCR. In an evaluation on nasopharyngeal aspirates from 166 patients with community-acquired pneumonia, the assay was positive in 49 of 50 culture-positive cases. Of 19 culture-negative but Spn9802 PCR-positive cases, 12 were confirmed as S. pneumoniae by rnpB sequence analysis. With an expanded reference standard, including culture and rnpB sequencing, Spn9802 had a sensitivity of 94% and a specificity of 98%. A cutoff for clinically significant positivity was 10(4) DNA copies/mL, giving 71% sensitivity and 100% specificity. In conclusion, Spn9802 real-time PCR is highly sensitive and specific. The quantification it provides enables differentiation between pneumococcal pathogenicity and commensalism. PMID- 17916423 TI - [Adoption from Haiti and lead poisoning]. PMID- 17916418 TI - Identification and sequencing of ESTs from the halophyte grass Aeluropus littoralis. AB - Aeluropus littoralis (Gouan) Parl. is a C4 perennial halophyte monocotyledonous plant belonging to the same family as wheat. Growing as weed in dry salty areas or marshes, it is salt-secreting, rhizomatous and is used as forage. It is diploid (2n=2X=14) and has a relative small genome of around 342 Mb. A. littoralis is highly salt-tolerant since this plant has the ability to secrete salt. Thus, A. littoralis has the potential to become an important genetic resource for biotechnological strategies to improve salt and drought tolerance in economically important crops such as wheat. We have constructed SSH (Suppression Subtractive Hybridization) cDNA libraries from root (RSD45) and leaf (LSD45) tissues of 45 days old plants grown in the presence of 300 mM NaCl. We have also constructed full-length cDNA library from 15 days old salt stressed (300 mM NaCl) roots (RSTL15). Sequencing revealed 25 and 42 independent transcripts from the RSD45 and LSD45 cDNA libraries respectively, in both cases this was less than 25% of the clones sequenced. In contrast, 425 (60%) of the clones from the RSTL15 library revealed independent transcripts. After comparison with protein databases using BlastX, 335 (68%) ESTs (Expressed Sequence Tag) were classified into putative known functions and unclassified proteins, 59 (12%) have homology only to unidentified homologous sequences. A total of 98 (20%) of the ESTs have no homologies to known sequences in the protein databases which can be considered as novel. PMID- 17916424 TI - [An excessive diagnosis of constipation in children complaining of abdominal pain in a pediatric emergency department: an example of practice patterns]. AB - Prescriptions of abdominal x-ray and diagnosis of constipation seemed too frequent in children evaluated for abdominal pain (AP) in a paediatric emergency unit. OBJECTIVE: To evaluate the prevalence of constipation in children with AP, to determine clinical and radiological signs related to this diagnosis, and to compare frequencies of clinical criteria of functional chronic constipation (FCC) in children with AP with those of controls (children with mild traumatisms without AP). METHODS: This prospective observational study included children older than 4 years of age consulting for AP. RESULTS: Among 196 patients with AP, 53% had a diagnosis of constipation on discharge (28% isolated and 25% associated with an other diagnosis), and 88% had a plain abdominal x-ray. A faecal retention was considered by emergency practitioners in 92% of children with the diagnosis of constipation vs 22% with other diagnosis (OR=38; CI 95%: 15-101). Clinical criteria of FCC were associated with the diagnosis of constipation (OR=2.7; CI 95%: 1.4-5.2). A FCC was as prevalent in the control population as in AP population (32 vs 34%). CONCLUSION: An excessive prevalence of diagnosis of constipation was mainly associated with contestable x-ray diagnosis of faecal retention secondary to inappropriate indications of abdominal x-ray. PMID- 17916425 TI - [Is there a risk with the skin-to-skin practice at birth?]. PMID- 17916427 TI - [Drug-facilitated crime and sexual abuse: a pediatric observation]. AB - Drug-facilitated crime in sexual assault situations remains insufficiently recognized by physicians. In the possible context of an assault and in front of recent neuropsychicological disturbances in a child, such an issue has to be considered. The quality of sampling, the use of ultra-sensitive and specific toxicologic methods and a clinical-biological collaboration allow to recognize this form of delinquency whose consequences are both medical and legal. PMID- 17916428 TI - [Childhood obesity: the necessity of a consensus on prevention]. PMID- 17916426 TI - [Persistent central diabetes insipidus in a very low birth weight infant]. AB - Central diabetes insipidus (DI) is extremely rare during the neonatal period. Most cases of central DI are secondary to a known aetiology. Substitutive treatment with desmopressin is effective with nasal or oral preparation, but doses are variable and must be tailored individually. We report on a case in a very low birth weight infant with an idiopathic central DI during the first month of life. He was successfully treated with oral desmopressin. The treatment was maintained after discharge with low doses desmospressin. PMID- 17916429 TI - Neurogenic responses to amyloid-beta plaques in the brain of Alzheimer's disease like transgenic (pPDGF-APPSw,Ind) mice. AB - Formation and accumulation of amyloid-beta (A beta) plaques are associated with declined memory and other neurocognitive function in Alzheimer's disease (AD) patients. However, the effects of A beta plaques on neural progenitor cells (NPCs) and neurogenesis from NPCs remain largely unknown. The existing data on neurogenesis in AD patients and AD-like animal models remain controversial. For this reason, we utilized the nestin second-intron enhancer controlled LacZ (pNes LacZ) reporter transgenic mice (pNes-Tg) and Bi-transgenic mice (Bi-Tg) containing both pPDGF-APPSw,Ind and pNes-LacZ transgenes to investigate the effects of A beta plaques on neurogenesis in the hippocampus and other brain regions of the AD-like mice. We chose transgenic mice at 2, 8 and 12 months of age, corresponding to the stages of A beta plaque free, plaque onset and plaque progression to analyze the effects of A beta plaques on the distribution and de novo neurogenesis of (from) NPCs. We demonstrated a slight increase in the number of NPCs in the hippocampal regions at the A beta plaque free stage, while a significant decrease in the number of NPCs at A beta plaque onset and progression stages. On the other hand, we showed that A beta plaques increase neurogenesis, but not gliogenesis from post-mitotic NPCs in the hippocampus of Bi-Tg mice compared with age-matched control pNes-Tg mice. The neurogenic responses of NPCs to A beta plaques suggest that experimental approaches to promote de novo neurogenesis may potentially improve neurocognitive function and provide an effective therapy for AD. PMID- 17916430 TI - Ameliorating effects of preadolescent aniracetam treatment on prenatal ethanol induced impairment in AMPA receptor activity. AB - Ethanol-induced damage in the developing hippocampus may result in cognitive deficits such as those observed in fetal alcohol spectrum disorder (FASD). Cognitive deficits in FASD are partially mediated by alterations in glutamatergic synaptic transmission. Recently, we reported that synaptic transmission mediated by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs) is impaired following fetal ethanol exposure. This finding led us to develop a rational approach for the treatment of alcohol-related cognitive deficits using aniracetam, an allosteric AMPAR modulator. In the present study, 28 to 34-day-old rats exposed to ethanol in utero were treated with aniracetam, and subsequently exhibited persistent improvement in mEPSC amplitude, frequency, and decay time. Furthermore, these animals expressed positive changes in synaptic single channel properties, suggesting that aniracetam ameliorates prenatal ethanol-induced deficits through modifications at the single channel level. Specifically, single channel open probability, conductance, mean open and closed times, and the number and burst duration were positively affected. Our findings emphasize the utility of compounds which slow the rate of deactivation and desensitization of AMPARs such as aniracetam. PMID- 17916432 TI - mGlu4 potentiation of K(2P)2.1 is dependant on C-terminal dephosphorylation. AB - Two-pore domain potassium (K(2P)) channels are proposed to underlie the background or leak current found in many excitable cells. Extensive studies have been performed investigating the inhibition of K(2P)2.1 by Galpha(q)- and Galpha(s)-coupled G-protein-coupled receptors (GPCRs), whereas in the present study we investigate the mechanisms underlying Galpha(i)/Galpha(o)-coupled GPCR increases in K(2P)2.1 activity. Activation of mGlu4 increases K(2P)2.1 activity, with pharmacological inhibition of protein kinases and phosphatases revealing the involvement of PKA whereas PKC, PKG or protein phosphatases play no role. Mutational analysis of potential C-terminal phosphorylation sites indicates S333 to control approximately 70%, with S300 controlling approximately 30% of the increase in K(2P)2.1 activity following mGlu4 activation. These data reveal that activation of mGlu4 leads to an increase in K(2P)2.1 activity through a reduction in C-terminal phosphorylation, which represents a novel mechanism by which group III mGlu receptors may regulate cell excitability and synaptic activity. PMID- 17916431 TI - Regulation of low affinity neurotrophin receptor (p75(NTR)) by early growth response (Egr) transcriptional regulators. AB - The low affinity neurotrophin receptor p75(NTR) is a multifunctional receptor with important roles in neurotrophin signaling, axon outgrowth, and oligodendroglia and neuron survival. It is transcriptionally regulated with spatial and temporal precision during nervous system development, injury and regeneration. Very little is known about how p75(NTR) expression is dynamically regulated but it is likely to influence how p75(NTR) signals in particular cellular contexts. Here, we identify the early growth response (Egr) transcriptional regulators, Egr1 and Egr3, as direct modulators of p75(NTR) gene expression. Egr1 and Egr3 bind and transactivate the p75(NTR) promoter in vitro and in vivo, using distinct response elements on the p75(NTR) promoter. Consistent with these results, p75(NTR) expression is greatly diminished in muscle spindle stretch receptors and in peripheral nerve Schwann cells in Egr gene deficient mice. Taken together, the results elucidate a novel mechanism whereby Egr proteins can directly modulate p75(NTR) expression and signaling in vivo. PMID- 17916434 TI - Transcriptional complexity of the HOXA9 locus. AB - Evolutionarily conserved HOX genes play an important role during development and hematopoiesis. HOX protein products are transcription factors whose precise mechanism of action is still poorly understood. Regulation of HOX gene expression has been the topic of various studies. While alternative splicing and alternative promoter usage have been known to increase the number of transcripts across the HOX clusters, more recently high-throughput analyses have identified a number of new coding and noncoding RNA molecules whose function is not known. Here we review the transcriptome of the most studied HOX locus, HOXA9. Strict control of HOXA9 expression has been shown to play a critical role in hematopoiesis while aberrant expression has been shown to be important to the development of leukemia. However, it is still unclear how various transcripts from this locus are regulated and what specific role(s) each one of them plays. PMID- 17916435 TI - Hematologically important mutations: Shwachman-Diamond syndrome. AB - Shwachman-Diamond syndrome (SDS) is a rare autosomal recessive disorder characterized by exocrine pancreatic insufficiency, bone marrow dysfunction, and skeletal abnormalities. The Shwachman-Bodian-Diamond syndrome (SBDS) gene was identified as a causative gene for SDS in 2003, and genetic analyses of SDS have been performed. Over the last 4 years, a number of different mutations affecting the SBDS gene have been described. In this report, a summary of documented SDS associated mutations is provided. PMID- 17916433 TI - Gephyrin clustering is required for the stability of GABAergic synapses. AB - Although gephyrin is an important postsynaptic scaffolding protein at GABAergic synapses, the role of gephyrin for GABAergic synapse formation and/or maintenance is still under debate. We report here that knocking down gephyrin expression with small hairpin RNAs (shRNAs) in cultured hippocampal pyramidal cells decreased both the number of gephyrin and GABA(A) receptor clusters. Similar results were obtained by disrupting the clustering of endogenous gephyrin by overexpressing a gephyrin-EGFP fusion protein that formed aggregates with the endogenous gephyrin. Disrupting postsynaptic gephyrin clusters also had transsynaptic effects leading to a significant reduction of GABAergic presynaptic boutons contacting the transfected pyramidal cells. Consistent with the morphological decrease of GABAergic synapses, electrophysiological analysis revealed a significant reduction in both the amplitude and frequency of the spontaneous inhibitory postsynaptic currents (sIPSCs). However, no change in the whole-cell GABA currents was detected, suggesting a selective effect of gephyrin on GABA(A) receptor clustering at postsynaptic sites. It is concluded that gephyrin plays a critical role for the stability of GABAergic synapses. PMID- 17916436 TI - Fatty acid composition of wild anthropoid primate milks. AB - Fatty acids in milk reflect the interplay between species-specific physiological mechanisms and maternal diet. Anthropoid primates (apes, Old and New World monkeys) vary in patterns of growth and development and dietary strategies. Milk fatty acid profiles also are predicted to vary widely. This study investigates milk fatty acid composition of five wild anthropoids (Alouatta palliata, Callithrix jacchus, Gorilla beringei beringei, Leontopithecus rosalia, Macaca sinica) to test the null hypothesis of a generalized anthropoid milk fatty acid composition. Milk from New and Old World monkeys had significantly more 8:0 and 10:0 than milk from apes. The leaf eating species G. b. beringei and A. paliatta had a significantly higher proportion of milk 18:3n-3, a fatty acid found primarily in plant lipids. Mean percent composition of 22:6n-3 was significantly different among monkeys and apes, but was similar to the lowest reported values for human milk. Mountain gorillas were unique among anthropoids in the high proportion of milk 20:4n-6. This seems to be unrelated to requirements of a larger brain and may instead reflect species-specific metabolic processes or an unknown source of this fatty acid in the mountain gorilla diet. PMID- 17916437 TI - [Degeneration of dermoid cysts: a case study of malignant transformation]. AB - Malignant transformation of ovarian dermoid cyst (mature cystic teratoma) is rare and most often established in postmenopausal women after surgery by sample anatomopathological analysis. We report the case of a 58-year-old woman showing abdominal pain associated with constipation episodes. The diagnosis of dermoid cyst was established upon ultrasonographic and tomodensitometric criteria. Its histopathological analysis confirmed the presence of a well differentiated epidermoid carcinoma. In line with the literature, our study highlights the importance of the age of the patient, the size of dermoid cyst and its growth rate as well as squamous cell carcinoma (SCC) antigen serum level. These date are collected in order to establish a correct diagnosis and provide an appropriate cure. When a cyst is discovered these data must pathological event, mostly observed in menopaused women. Certainty diagnosis is collected in order to establish a correct diagnosis and provide an appropriate cure. PMID- 17916438 TI - [Abruptio placentae. Management in a reference Nigerien maternity. Prospective study about 118 cases during one year]. AB - OBJECTIVE: Placental abruption is a syndrome, which occurs in the third trimester of the pregnancy or during labour. It is the main cause of pregnancy last term bleeding and is also responsible for a high stillbirth rate. The objective is to describe the epidemiological, clinical, paraclinical and therapeutic characteristics in order to decrease the fetal and maternal mortality and morbidity. PATIENTS AND METHODS: It is a prospective and descriptive study in continuous series, over a twelve-month period, from 1st January 2003 to 31st December 2003, at Issaka Gazobi maternity of Niamey, Niger. RESULTS: During the study period, 3255 deliveries have been done. One hundred and eighteen placental abruptions have been observed, which corresponds to a frequency of 3.6% with the highest rate during raining season. The average age and parity were: 31 years and 5th with a predominance of grand multiparous (38.2%). The majority of the patients were in-utero transfers (83,1%) and had done at least 10 km before arriving. One hundred and eleven patients had a caesarean section (94,1%) and 7 delivered through the vaginal route (5.9%). The fetal prognosis was characterized by a high stillbirth rate of 71.3% (87/122), fetal hypotrophy (64.8%) and prematurity (23.8%). The main cause of maternal morbidity was anaemia (76.3%) and Disseminated Intravascular Coagulation (5.9%). Further, 81 patients have been transfused (68.7%). The maternal mortality was 5.1% (6/118). DISCUSSION AND CONCLUSION: Placental abruption, a medical and obstetrical emergency, is a serious obstetric condition, especially in our country. Physicians must be aware that patients with high blood pressure, preeclampsia, eclampsia, particularly in case of multiparity are at increased risk of placental abruption. Early diagnosis, prenatal follow-up and caesarean section improve the maternal and fetal prognosis. PMID- 17916439 TI - [Increased risk of relapse in multiple sclerosis patients after ovarian stimulation for in vitro fertilization]. AB - In this preliminary study we analysed the impact of ovarian stimulations and the different protocols used for in vitro fertilizations (IVF) on the clinical activity of multiple sclerosis (MS). By matching the databases on MS and IVF of the past 10 years at the university hospital of Nantes, six patients have been found and, for five of them MS relapse rate seemed to be increased in the three month period following IVF as compared to the previous three months and to two other control periods of three months (P<0.05, Friedman test). The increased relapse rate mainly concerned patients treated by GnRH agonists but not the patients treated by GnRH antagonists. This preliminary work suggests a possible impact of the treatments used for IVF on MS relapse rate. Further studies are now underway to validate these results on a larger scale, by including all cases reported in France. PMID- 17916440 TI - Soluble CD14 and toll-like receptor-2 are potential salivary biomarkers for oral lichen planus and burning mouth syndrome. AB - Oral lichen planus (OLP) and burning mouth syndrome (BMS) are chronic conditions affecting the oral mucosa characterized by pain and burning sensation. Saliva plays a significant role in the maintenance of physical and functional integrity of normal oral mucosa. Identification of potential "salivary biomarkers" for early diagnosis and/or monitoring of human diseases is being explored. We investigated the soluble forms of innate immune associated proteins CD14 and toll like receptor-2 in unstimulated whole saliva (UWS) as potential biomarkers for OLP and BMS. Our results suggest that the levels of sCD14 and sTLR-2 in UWS were upregulated in OLP and BMS respectively. In addition, oral epithelial cells in the saliva of patients with OLP and BMS exhibited elevated levels of CD14 mRNA and decreased levels of TLR-2 mRNA. Interestingly, presence of co-existent oral candidiasis nullified these changes. PMID- 17916441 TI - Immunophenotypic alterations in acute and early HIV infection. AB - To understand the extent of immune dysregulation in primary HIV infection (PHI) and the impact of antiretroviral therapy (ART) on restoring these abnormalities, we longitudinally evaluated 52 subjects (Acute-Treated (AT); Early-Treated (ET); Early Untreated (EU)) for markers of activation, proliferation, and function on T cells. ET and AT patients differed by 0.54 log viral load (VL) at baseline but did not differ thereafter by more than 0.34 log10 VL. AT subjects had higher CD8(+) T cell counts and expression of markers indicative of CD8(+) T cell activation (CD38), and proliferation (Ki67), at baseline, than ET subjects but were not different 48 weeks post-ART. Although acute PHI is marked by higher level of immune activation than early PHI, virologic and immunologic responses were similar post-ART, suggesting that the extent of immunologic recovery is not negatively impacted by a delay of treatment beyond the acute stage of disease. PMID- 17916442 TI - Chronic innate immune activation as a cause of HIV-1 immunopathogenesis. AB - Human immunodeficiency virus (HIV)-1 infection causes progressive impairment of the immune system in humans, characterized by depletion of CD4 T cells and loss of T cell function. Increased markers of T cell activation and lymphoid hyperplasia suggest that chronic T cell activation persists in immunocompromised hosts, and contributes to the exhaustion of immune functions. Here we propose a revision of this hypothesis, in which we suggest that chronic activation of innate immunity may negatively affect adaptive T cell-mediated responses. We hypothesize that constant exposure of the effector cells of innate immunity to HIV results in their chronic hyperactivation, with deleterious effects on T cells. In particular, plasmacytoid dendritic cells (pDC) may be highly susceptible to HIV-induced activation due to its interaction with the cellular receptor CD4, expressed by pDC. Subsequent production of type I interferon and indoleamine 2,3-dioxygenase may exert suppressive and cytotoxic effects on T cells. PMID- 17916443 TI - Human thymic dendritic cells: regulators of T cell development in health and HIV 1 infection. AB - Thymic dendritic cells (DCs) are a unique subset of bone marrow-derived professional antigen presenting cells (APCs) that interact closely with developing thymocytes and play a crucial role in the process of negative selection and subsequent deletion of potential auto-reactive T cell clones. HIV-1 infection of the thymus has been implicated in the defective regeneration of the CD4(+) T cell pool in infected individuals. Thymic DCs are permissive to infection by HIV-1 and given their important role in T cell development, infected DCs within the thymus may contribute to the depletion of T cells. Here we review the phenotype and function of different DC subsets found within the human thymus and discuss potential mechanisms of how DCs may be important in CD4(+) T cell dysfunction in HIV-1 infection. PMID- 17916446 TI - Developmental changes of FOXP3-expressing CD4+CD25+ regulatory T cells and their impairment in patients with FOXP3 gene mutations. AB - FOXP3 is required for the generation and function of CD4(+)CD25(+) regulatory T (Treg) cells. To elucidate the biological role of Treg cells, we used a monoclonal anti-FOXP3 antibody to examine the frequencies of Treg cells during child development. The percentages of CD4(+)CD25(+)FOXP3(+) T cells were constant shortly from after birth through adulthood. CD4(+)CD25(+)FOXP3(+) T cells in cord blood showed the naive CD45RA(+)CD45RO(-) phenotype, whereas adult CD4(+)CD25(+)FOXP3(+) T cells expressed mostly the memory CD45RA(-)CD45RO(+) phenotype. The age-dependent dominance of memory CD4(+)CD25(+)FOXP3(+) T cells implies functional differences between naive and memory Treg cells. Notably, four patients with FOXP3 gene mutations revealed a paucity of CD4(+)CD25(+)FOXP3(+) T cells. Importantly, one patient with a frame shift mutation, who showed typical symptoms of IPEX (immune dysregulation, polyendocrinopathy, enteropathy, X linked), exhibited marked T cell activation, whereas others with missense mutations, who were clinically milder, did not. This observation suggests a possible genotype-phenotype correlation in IPEX. PMID- 17916447 TI - Phase I/II study of immunotherapy using autologous tumor lysate-pulsed dendritic cells in patients with metastatic renal cell carcinoma. AB - This phase I/II study was conducted to evaluate the feasibility, safety and efficacy of immunotherapy using tumor lysate (TL)-pulsed dendritic cells (DC) in patients with metastatic renal cell carcinoma (RCC). DC were generated by culturing peripheral blood mononuclear cells in the presence of GM-CSF and IL-4 and were pulsed with autologous TL and keyhole limpet hemocyanin (KLH). Maturation of DC was induced by a combined treatment of CD40 ligand, IFN and monocyte-conditioned medium. The patients were administered two cycles of TL pulsed DCs vaccination, each of which comprised of four doses injected subcutaneously at biweekly intervals. Nine patients were included. The immunotherapy was well tolerated without severe side effects. One patient achieved an objective partial response (PR). Five patients showed stable disease (SD), and the remaining three had progressive disease (PD). With a median follow up of 17.5 months, the median time to progression was 5.2 months and the median overall survival was 29 months. In the antigen-specific lymphocyte proliferation assay, eight patients showed a proliferative response, which tended to be stronger in patients with SD or PR than in patients with PD. The ELISPOT assay was performed in two patients and indicated that one patient with PR showed a much stronger response than another with PD. Our results suggest that TL-pulsed DC immunotherapy in combination with nephrectomy affect the natural course of RCC and are associated with clinical benefits for patients with metastatic diseases. PMID- 17916444 TI - TNFalpha blockade in human diseases: mechanisms and future directions. AB - Tumor necrosis factor-alpha (TNFalpha) antagonists have shown remarkable efficacy in a variety of immune-mediated inflammatory diseases (IMIDs). Therapeutic scope and limitations of these agents are reviewed in a recently published article in the Journal. In spite of their therapeutic popularity, little is known about their modes of action in vivo and factors that limit their scope of therapeutic use. Intriguingly, while all TNFalpha antagonists including blocking antibodies and soluble receptors are effective in certain IMIDs, only anti-TNFalpha antibodies are effective in other IMIDs. Early efforts at understanding how TNFalpha antagonists act in IMIDs centered on their ability to neutralize soluble TNFalpha or to block TNF receptors from binding to their ligands. Subsequent studies suggested a role of complement-mediated lysis or antibody-dependent cell cytotoxicity in their therapeutic effects. More recent models postulate that TNFalpha blockers may act by affecting intracellular signaling, with the end result being a hastened cell cycle arrest, apoptosis, suppression of cytokine production, or improved Treg cell function. TNFalpha antagonists can also modulate the functions of myofibroblasts and osteoclasts, which might explain how TNFalpha antagonists reduce tissue damage in chronic IMIDs. Focusing on the human therapeutic experience, this analytical review will review the biology of mechanisms of action, the limiting factors contributing to disease restriction in therapeutic efficacy, and the mechanism and frequency of treatment-limiting adverse responses of TNFalpha antagonists. It is hoped that the overview will address the needs of clinicians to decide on optimal use, spur clinical innovation, and incite translational researchers to set priorities for in vivo human investigations. PMID- 17916445 TI - TNFalpha blockade in human diseases: an overview of efficacy and safety. AB - Tumor necrosis factor-alpha (TNFalpha) antagonists including antibodies and soluble receptors have shown remarkable efficacy in various immune-mediated inflammatory diseases (IMID). As experience with these agents has matured, there is an emerging need to integrate and critically assess the utility of these agents across disease states and clinical sub-specialties. Their remarkable efficacy in reducing chronic damage in Crohn's disease and rheumatoid arthritis has led many investigators to propose a new, 'top down' paradigm for treating patients initially with aggressive regimens to quickly control disease. Intriguingly, in diseases such as rheumatoid arthritis and asthma, anti-TNFalpha agents appear to more profoundly benefit patients with more chronic stages of disease but have a relatively weaker or little effect in early disease. While the spectrum of therapeutic efficacy of TNFalpha antagonists widens to include diseases such as recalcitrant uveitis and vasculitis, these agents have failed or even exacerbated diseases such as heart failure and multiple sclerosis. Increasing use of these agents has also led to recognition of new toxicities as well as to understanding of their excellent long-term tolerability. Disconcertingly, new cases of active tuberculosis still occur in patients treated with all TNFalpha antagonists due to lack of compliance with recommendations to prevent reactivation of latent tuberculosis infection. These safety issues as well as guidelines to prevent treatment-associated complications are reviewed in detail in this article. New data on mechanisms of action and development of newer TNFalpha antagonists are discussed in a subsequent article in the Journal. It is hoped that these two review articles will stimulate a fresh assessment of the priorities for research and clinical innovation to improve and extend therapeutic use and safety of TNFalpha antagonism. PMID- 17916448 TI - Vestigial-like-2b (VITO-1b) and Tead-3a (Tef-5a) expression in zebrafish skeletal muscle, brain and notochord. AB - The vestigial gene has been shown to control skeletal muscle formation in Drosophila and the related Vestigial-like 2 (Vgl-2) protein plays a similar role in mice. Vgl-family proteins are thought to regulate tissue-specific gene expression by binding to members of the broadly expressed Scalloped/Tef/TEAD transcription factor family. Zebrafish have at least four Vgl genes, including two Vgl-2s, and at least three TEAD genes, including two Tead3s. We describe the cloning and expression of one member from each family in the zebrafish. A novel gene, vgl-2b, with closest homology to mouse and human vgl-2, is expressed transiently in nascent notochord and in muscle fibres as they undergo terminal differentiation during somitogenesis. Muscle cells also express a TEAD-3 homologue, a possible partner of Vgl-2b, during myoblast differentiation and early fibre assembly. Tead-3a is also expressed in rhombomeres, eye and epiphysis regions. PMID- 17916449 TI - Prevalence and genetic diversity of simian immunodeficiency virus infection in wild-living red colobus monkeys (Piliocolobus badius badius) from the Tai forest, Cote d'Ivoire SIVwrc in wild-living western red colobus monkeys. AB - Numerous African primates are infected with simian immunodeficiency viruses (SIVs). It is now well established that the clade of SIVs infecting west-central African chimpanzees (Pan troglodytes troglodytes) and western gorillas (Gorilla gorilla gorilla) represent the progenitors of human immunodeficiency virus type 1 (HIV-1), whereas HIV-2 results from different cross-species transmissions of SIVsmm from sooty mangabeys (Cercocebus atys atys). We present here the first molecular epidemiological survey of simian immunodeficiency virus (SIVwrc) in wild-living western red colobus monkeys (Piliocolobus badius badius) which are frequently hunted by the human population and represent a favourite prey of western chimpanzees (Pan troglodytes verus). We collected faecal samples (n=88) and we assessed individual discrimination by microsatellite analyses and visual observation. We tested the inferred 53 adult individuals belonging to two neighbouring habituated groups for presence of SIVwrc infection by viral RNA (vRNA) detection. We amplified viral polymerase (pol) (650 bp) and/or envelope (env) (570 bp) sequences in 14 individuals, resulting in a minimal prevalence of 26% among the individuals sampled, possibly reaching 50% when considering the relatively low sensitivity of viral RNA detection in faecal samples. With a few exceptions, phylogenetic analysis of pol and env sequences revealed a low degree of intragroup genetic diversity and a general viral clustering related to the social group of origin. However, we found a higher intergroup diversity. Behavioural and demographic data collected previously from these communities indicate that red colobus monkeys live in promiscuous multi-male societies, where females leave their natal group at the sub-adult stage of their lives and where extra-group copulations or male immigration have been rarely observed. The phylogenetic data we obtained seem to reflect these behavioural characteristics. Overall, our results indicate that wild-living red colobus represent a substantial reservoir of SIVwrc. Moreover, because of their frequent association with other monkey species, the predation pressure exerted by chimpanzees (Pan troglodytes verus) and by poachers around and inside the park, simian to simian and simian to human SIVwrc cross-species transmission cannot be excluded. PMID- 17916450 TI - Hemocyanin conformational changes associated with SDS-induced phenol oxidase activation. AB - The enzymatic activity of phenoloxidase is assayed routinely in the presence of SDS. Similar assay conditions elicit phenoloxidase activity in another type 3 copper protein, namely hemocyanin, which normally functions as an oxygen carrier. The nature of the conformational changes induced in type 3 copper proteins by the denaturant SDS is unknown. This comparative study demonstrates that arthropod hemocyanins can be converted from being an oxygen carrier to a form which exhibits phenoloxidase activity by incubation with SDS, with accompanying changes in secondary and tertiary structure. Structural characterisation, using various biophysical methods, suggests that the micellar form of SDS is required to induce optimal conformational transitions in the protein which may result in opening a channel to the di-copper centre allowing bulky phenolic substrates access to the catalytic site. PMID- 17916451 TI - Effect of salt on the activity of Streptomyces prolyl aminopeptidase. AB - A salt-tolerant prolyl aminopeptidase from Streptomyces aureofaciens TH-3 (TH 3PAP) was purified from a culture supernatant. The gene encoding TH-3PAP was cloned and sequenced. The primary structure of TH-3PAP showed 65% identity with that of PAP from Streptomyces lividans (SLPAP) and possessed a conserved catalytic motif, GxSxGG, which is conserved in the alpha/beta hydrolase fold family. The characterization of the recombinants TH-3PAP and SLPAP indicated a difference: in 4.0 M NaCl, TH-3PAP showed enzyme activity, whereas SLPAP was inactive. Next, we constructed chimeras between TH-3PAP and SLPAP using an in vivo DNA shuffling system and a sandwich chimera (sc-PAP), whose region from 63 to 78 amino acids of TH-3PAP was substituted with that of SLPAP. Comparison of the biochemical properties between TH-3PAP and the salt-sensitive sc-PAP suggested that the fine tuning of the N-terminal conformation of TH-3PAP by hydrophobic interaction is important for the salt tolerance mechanism of the enzyme. PMID- 17916452 TI - Increased cathepsin K and tartrate-resistant acid phosphatase expression in bone of streptozotocin-induced diabetic rats. AB - The effect of insulin-dependent diabetes mellitus (IDDM) on bone metabolism was evaluated using the streptozotocin (STZ)-induced diabetic rat 1 week after the induction of diabetes. The urinary excretion of cross-linked N-telopeptides of type I collagen (NTx) and deoxypyridinoline (Dpd) in diabetic rats increased to 3.6-fold and 1.2-fold the control level, respectively. The amount of hydroxyproline and calcium in the distal femur of diabetic rats significantly decreased to 76% and 90% of the control, respectively. The levels of serum osteocalcin and alkaline phosphatase (ALP) activity in the distal femur of the diabetic rats were significantly reduced to about 40% and 70% of the control levels, respectively. The decrease in the expression osteocalcin was observed in distal femur of the diabetic rats, although the level of ALP mRNA was unchanged. The activity and the mRNA level of tartrate-resistant acid phosphatase (TRAP) increased to 1.5- and 2.3-fold the control level, respectively, in distal femur of the diabetic rats. The activity, protein, and mRNA levels of cathepsin K of diabetic rats also elevated to about 2-, 2.3-, and 2-fold the control levels, respectively. These results suggest that IDDM contributes to bone loss through changes in gene expression of TRAP and cathepsin K in osteoclasts as well as osteocalcin in osteoblasts resulting in increased bone resorptive activity and decreased bone formation. PMID- 17916454 TI - Inferior vena cava filters in the management of venous thromboembolism. AB - Inferior vena cava (IVC) filters, both retrievable and permanent, are indicated for the prevention of pulmonary embolism (PE) in patients contraindicated for anticoagulant therapy, in those with anticoagulant therapy complications, and perhaps for those with recurrent PE despite therapeutic anticoagulation. Because of the lack of randomized controlled trials (only 1 has been published), clinicians have little evidence-based information to assist them in determining proper use of IVC filters. The introduction of retrievable filters and the ease of insertion have stimulated increased use of these devices without strong evidence or follow-up to assess either efficacy or longer-term clinical outcomes. Current evidence-based guidelines recommend IVC filter insertion only in patients with proven venous thromboembolism and an absolute contraindication for anticoagulation. PMID- 17916455 TI - Outpatient management of stable acute pulmonary embolism: proposed accelerated pathway for risk stratification. AB - Pulmonary embolism (PE) is a major health problem and a cause of worldwide morbidity and mortality. The current standard therapy for acute PE encourages admitting patients to the hospital for administration of parenteral anticoagulation therapy as a bridge to oral vitamin K antagonists. Prognostic models that identify patients with stable (nonmassive) acute PE (SPE) who are at low risk for adverse outcome have recently been reported. Based on these risk stratification models, hospital-based therapy is warranted for patients with PE who meet the criteria associated with a high risk for adverse outcome. However, a growing body of evidence suggests the feasibility of partial outpatient management and accelerated hospital discharge (AHD) in a subset of patients with SPE. Prospective validation of these risk stratification models for predicting patient suitability for AHD is needed. PMID- 17916456 TI - Newer modalities for detection of pulmonary emboli. AB - Pulmonary embolism (PE) is the third most common cardiovascular disease after myocardial infarction and stroke in the United States. Early and accurate diagnosis of this condition is imperative because many patients die within hours of presentation. Clinical and laboratory tests can be used to accurately determine the pretest probability of PE. When necessary, imaging techniques are then used to exclude or diagnose PE. Pulmonary angiography is the reference standard for the diagnosis of PE, but it is invasive and has a high morbidity and mortality rate. Ventilation and perfusion (V/Q) scanning in the past has been recommended as the initial diagnostic test for PE; however, this technique also has limitations. Recently, new modalities for the diagnosis and exclusion of PE have been evaluated. These techniques include V/Q single photon emission computed tomography (SPECT), single- and multi-detected computed tomography, and magnetic resonance angiography (MRA) including gadolinium-enhanced MRA, real-time magnetic resonance imaging (RT-MR), and magnetic resonance perfusion imaging. PMID- 17916457 TI - Prevention and management of venous thromboembolism in pregnancy. AB - Normal pregnancy is accompanied by an increase in clotting factors. The resulting hypercoagulable state has likely evolved to protect women from hemorrhage at the time of miscarriage and childbirth. During pregnancy, women are 4 times more likely to suffer from venous thromboembolism (VTE) compared with when they are not pregnant. Relative to pregnancy, the risk postpartum is even higher. The incidence of VTE is approximately 2 per 1,000 births, and VTE accounts for 1 death per 100,000 births, or approximately 10% of all maternal deaths. The most important risk factors during pregnancy are thrombophilia and a history of thrombosis. A history of thrombosis increases the risk for VTE to 2% to 12%. Thrombophilia increases not only the risk for maternal thrombosis but also the risk of poor pregnancy outcome. Despite the increased risk for thrombosis during pregnancy and the postpartum period, most women do not require anticoagulation. Those who do require anticoagulation include women with current VTE, women on lifelong anticoagulation, and many women with thrombophilia or a history of thrombosis. Recommended options for anticoagulation in pregnancy are limited to heparins, which do not cross the placenta. Low-molecular-weight heparin (LMWH) is preferred over unfractionated heparin because LMWH has a longer half-life and is presumed to have fewer side effects. The longer half-life is a disadvantage around the time of delivery, when unfractionated heparin, with its shorter half life, is easier to manage. For women who develop or are at high risk for heparin induced thrombocytopenia or severe cutaneous reactions, fondaparinux is probably the agent of choice. Women who do not require lifelong anticoagulation, but require anticoagulation during pregnancy, will still require anticoagulation for the first 6 weeks postpartum. PMID- 17916458 TI - The pharmacoeconomics of deep vein thrombosis treatment. AB - Venous thromboembolism (VTE), encompassing both deep vein thrombosis (DVT) and pulmonary embolism, remains a common and costly condition that is associated with significant morbidity and mortality. Treatment options for initial management of DVT include unfractionated heparin (UFH), low-molecular-weight heparins (LMWHs), and fondaparinux, which is the first of a new class of pentasaccharide antithrombotic agents with anti-factor Xa activity. LMWHs are an important tool in DVT management, offering advantages over UFH such as ease of dosing, lack of need for coagulation monitoring, and reduced risk for heparin-induced thrombocytopenia (HIT). Fondaparinux is also characterized by a simple dosing regimen, no need for coagulation monitoring, and potentially a lower risk of HIT compared with LMWH. In a recent clinical trial of DVT management, efficacy and bleeding rates with fondaparinux appeared similar to those observed with LMWH. In contrast to LMWH, fondaparinux is generally given as a fixed dose across a range of patient weights rather than calculated per individual patient weight. Given the increasing economic burden of VTE, particularly due to its increased rate among the elderly, pharmacoeconomic analyses have become a particularly useful tool to aid in selecting among similarly effective and safe agents for VTE treatment. A recent cost-effective analysis demonstrated that fondaparinux use offers an attractive economic alternative to other agents for initial DVT therapy that could yield cost savings without compromising clinical outcomes or patient safety. PMID- 17916459 TI - Neuroplasticity as a target for the pharmacotherapy of psychiatric disorders: new opportunities for synergy with psychotherapy. PMID- 17916460 TI - Editorial. PMID- 17916461 TI - Clinical proteomics: discovery of cancer biomarkers using mass spectrometry and bioinformatics approaches--a prostate cancer perspective. AB - Prostate cancer (PCa) is an intractable disease, where diagnosis and clinical prediction of the disease course and response to treatment is compromised by the lack of objective and robust biomarker assays. In late stage metastatic disease, treatment options are limited, although it is recognized that some patients may benefit from immunotherapy and in particular vaccine therapy. However, research into biomarkers that correlate with the clinical outcome of immunotherapy has lagged behind vaccine development. Thus, proteomic tools are increasingly being utilized for the discovery of biomarkers which will allow us to make clinical decisions about patient treatment at an earlier stage and should aid in shortening the development time for vaccines. In this review we will summarize the various proteomic platforms used to investigate new biomarkers in PCa for better patient diagnosis, prognosis, patient stratification, treatment monitoring and clinical surrogate endpoints. We will discuss method limitations and highlight the key areas of research required for understanding the etiology of PCa. PMID- 17916462 TI - Cell based cancer vaccines: regulatory and commercial development. AB - There is both clinical and regulatory drive to expedite development of safe, efficacious cancer therapies. Stimulation of the patients immune system through vaccination with tumour cells has long been at the vanguard of cancer therapeutic vaccines, and several have been demonstrated to be safe and to have efficacy in early clinical trials for a range of cancers including melanoma, renal cell carcinoma, prostate and colorectal cancers. A number of development-stage vaccines and strategies are currently being tested, utilising either autologous or allogeneic tumour cells, which may also have been ex vivo manipulated (e.g. cytokine transfected cells). It seems likely that clinical trial success, and hence patient benefit, could be improved through better patient identification, possibly by the discovery and use of novel immune response biomarkers. In this review, we aim to summarise the state of tumour cell vaccines in commercial development and to explore not only the difficulties of determining efficacy, but also the production challenges faced when developing a vaccine from proof of principle to pivotal phase III trials. PMID- 17916463 TI - GSK's antigen-specific cancer immunotherapy programme: pilot results leading to Phase III clinical development. AB - From the first evidence that the immune system could recognize tumors, different types of tumor antigens have been identified and deeply characterized. Several different approaches aimed at targeting these antigens have already been the subject of clinical studies. In this field, the GSK Biologicals' approach relying on recombinant proteins combined with an immunological Adjuvant System in a specific clinical setting, has entertained hopes of developing a new class of well tolerated anti-cancer therapy. This methodology led to promising advances with MAGE-A3 immunotherapy in NSCLC and has the potential to be applied to all tumor types. PMID- 17916464 TI - Enhancing cancer vaccines with immunomodulators. AB - Harnessing the immune system to control cancer has been a challenge for cancer immunotherapists for many years. However, while specific immune responses to tumour-associated antigenic targets have been successfully induced in some patients, these responses have not always been sufficient to reproducibly and consistently mediate useful anti-tumour clinical activity. Many checks and balances have been incorporated into the immune response by nature to prevent or reduce the likelihood of autoimmunity or exaggerated protective inflammatory responses. Tolerance to self-antigens expressed on tumours is a major limitation in generating functional anti-tumour responses. In recent years, many of the pathways that mediate this tolerance have been identified, and reagents that can be used to manipulate these pathways have been clinically evaluated. These include: (a) pathways to activate professional antigen presenting cells, such as through Toll-like receptors, growth factors, such as GM-CSF, and the CD40 pathway; (b) use of cytokines, such as IL-2, IL-12, and Interferon alpha to enhance immune activation; and (c) pathways that inhibit T cell inhibitory signals, or Tregs. This article reviews clinical trials that have evaluated these approaches, and highlights promising combination vaccine/immunomodulator combination treatments based upon published clinical trial results. PMID- 17916465 TI - Lessons from randomized phase III studies with active cancer immunotherapies- outcomes from the 2006 meeting of the Cancer Vaccine Consortium (CVC). AB - After years of effort to develop active cancer immunotherapies, seven candidate products achieved promising results in phase I/II studies that triggered phase III randomized studies. One candidate to date has received an approvable letter from the United States Food and Drug Administration (FDA), defining a clear path to licensure for sipuleucel-T (Provenge, Dendreon) within the next couple of years. The other phase III studies failed to achieve statistical criteria for some or all of the critical endpoints. Yet, there is widespread recognition that using a patient's own immune system to target and destroy cancer cells may offer an effective biological therapy with less toxicity than presently available anti cancer therapies, and several candidates are still being evaluated in clinical studies. This review summarizes the lessons learned from these case studies, evaluates scientific, study design, and business factors that can affect study outcomes, identifies common challenges faced by sponsors developing these innovative therapies, and provides considerations for future study designs that may increase the likelihood of success. PMID- 17916467 TI - Results of modified Latarjet reconstruction in patients with anteroinferior instability and significant bone loss. AB - PURPOSE: The purpose of this study was to analyze the results of the modified Latarjet procedure for shoulder instability associated with an inverted-pear glenoid (bone loss of at least 25% of the width of the inferior glenoid) or an engaging Hill-Sachs lesion. METHODS: From March 1996 to December 2002, 102 patients underwent an open Latarjet procedure for shoulder instability with an inverted-pear glenoid, with or without an associated engaging Hill-Sachs lesion, by the 2 senior authors (S.S.B. and J.F.D.), and 47 of them were available for follow-up physical examination. The remaining 55 patients were contacted by telephone or letter to see if they had had recurrent dislocation or subluxation. The mean age of the patients was 26.5 +/- 6.6 years (range, 16 to 41 years). There were 46 male patients and 1 female patient. Preoperatively, mean forward elevation was 177.2 degrees +/- 13.6 degrees (range, 90 degrees to 180 degrees) and mean external rotation with the arm at the side was 55.3 degrees +/- 16.1 degrees (range, 0 degrees to 80 degrees). All patients had a positive apprehension sign preoperatively. The median number of dislocations before surgery was 6, with 20 patients having had more than 15 dislocations preoperatively. RESULTS: The mean follow-up time for the 47 patients who were personally examined was 59.0 +/- 18.5 months (range, 32 to 108 months). Postoperatively, mean forward elevation was 179.6 degrees +/- 2.0 degrees (range, 170 degrees to 180 degrees; gain of 2.4 degrees) and external rotation with the arm at the side was 50.2 degrees +/- 12.6 degrees (range, 22 degrees to 78 degrees; loss of 5.1 degrees). As for postoperative functional scores, the mean Constant score was 94.4 and the mean Walch-Duplay score was 91.7. None of these 47 patients showed any further dislocation, and 1 of them still had a positive apprehension sign (2.2%) indicating subluxation. However, 4 patients out of the total 102 who underwent the modified Latarjet procedure had a recurrence. With 4 recurrent dislocations and 1 recurrent subluxation, there was a 4.9% recurrence rate. The 4 patients with recurrent dislocations were not among the 47 who returned for personal follow-up evaluation. CONCLUSIONS: The 2 senior authors (S.S.B. and J.F.D.) have previously reported an unacceptably high recurrence rate (67%) for arthroscopic Bankart repair in the presence of an inverted-pear glenoid with or without an engaging Hill-Sachs lesion. They have recommended an open modified Latarjet procedure in such patients. The present study confirms the validity of that recommendation, because the same 2 surgeons have had only a 4.9% recurrence rate in that same category of patient at a mean follow-up of 59 months. Furthermore, the results of this study show the efficacy of the modified Latarjet procedure in the extremely challenging category of patients who present with such dramatic bone loss that soft-tissue reconstruction, either open or arthroscopic, is not a reasonable option. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17916468 TI - Radiofrequency-based plasma microtenotomy compared with arthroscopic subacromial decompression yields equivalent outcomes for rotator cuff tendinosis. AB - PURPOSE: This study aimed to determine whether radiofrequency (RF)-based plasma microtenotomy (microdebridement) was effective for treating chronic supraspinatus tendinosis. METHODS: The institutional ethics committee approved the study design, and all patients signed informed consent forms. Patients (age range, 30 to 70 years) were considered for enrollment if 6 months of active conservative treatment had failed and they had Neer stage II impingement syndrome, positive radiographic evidence of type II acromion, and magnetic resonance imaging or ultrasound evidence of supraspinatus tendinosis. Patients (N = 60) were randomly assigned to undergo arthroscopic subacromial decompression or RF-based plasma microtenotomy. For microtenotomy, a bipolar RF-based probe (TOPAZ; ArthroCare, Austin, TX) was used to perform microdebridement in the supraspinatus tendon; patients did not undergo acromioplasty. Outcomes evaluation consisted of self reported pain via a visual analog scale, as well as functional assessment (American Shoulder and Elbow Surgeons [ASES] survey, Constant score, and University of California, Los Angeles [UCLA] questionnaire). Statistical analyses were performed by use of factorial dependent-measures analysis of variance tests. RESULTS: Age and baseline scores on the visual analog scale (mean +/- SD) were 52.0 +/- 6.7 and 53.2 +/- 6.6 years and 8.4 +/- 0.9 and 8.2 +/- 0.8 points in the microtenotomy and arthroscopic subacromial decompression groups, respectively. A significant reduction in pain (P < .001) and improved function (P < .001 for all measures) were observed in both groups postoperatively. Both treatment groups had almost identical longitudinal recovery profiles for pain relief (P = .416) and restoration of function (P = .964 for ASES score, P = .978 for Constant score, and P = .794 for UCLA score). At 1 year, the median pain score was 1.0, and all patients had ASES, Constant, and UCLA scores of greater than 90, greater than 80, and greater than 30, respectively. CONCLUSIONS: Both procedures were associated with significant improvement postoperatively, but the RF-based plasma microtenotomy procedure draws into question the need for a more extensive procedure such as subacromial decompression in this patient population. LEVEL OF EVIDENCE: Level I, therapeutic randomized controlled study. PMID- 17916469 TI - Frequency of various tear patterns in full-thickness tears of the rotator cuff. AB - PURPOSE: We define specific rotator cuff tear patterns, prospectively document their occurrences, and emphasize the importance of tear pattern recognition during arthroscopy. METHODS: We prospectively analyzed 193 full-thickness rotator cuff tears intraoperatively from a single surgeon's practice. We address specific tear patterns, tear size, tissue mobility, tissue quality, and chronicity. RESULTS: Most of the tears were able to be categorized into 6 morphologic patterns. Of the tears, 3% could not be categorized. Appreciation of various rotator cuff tear patterns guided the positioning of bone and tissue sutures to achieve accurate apposition of tendon fibers. Transverse tears were the smallest and most common tear pattern, whereas tongue-shaped and U-shaped tears were larger patterns of comparable size. U-shaped tears had more retraction and less mobility and were of poorer tissue quality. Transverse tears were easily repaired, whereas U-shaped tears could not be repaired in 38% of cases (5/13). CONCLUSIONS: The quality and mobility of rotator cuff tissue were correlated with tear pattern, size, retraction, and chronicity. We describe a comprehensive rotator cuff tear classification scheme that encompasses 97% of all tears. Tear type was correlated with tendon retraction, tear size, cuff mobility, and tissue quality. On the basis of this information, the surgeon can anticipate tear patterns, which may improve pattern recognition and facilitate anatomic repair. LEVEL OF EVIDENCE: Level IV, prognostic case series. PMID- 17916470 TI - Arthroscopic-assisted treatment of coronoid fractures. AB - PURPOSE: Little information exists regarding arthroscopic treatment of coronoid fractures; this study reports outcomes in a series of patients. METHODS: Forty one coronoid fractures were identified by retrospective chart review; 7 were treated arthroscopically; 6 with >12 months of available follow-up. Regan-Morrey fracture types were II (n = 4) and III (n = 3). The age of the patients averaged 37 years; the mechanism of injury was a fall in 6 patients and a motocross accident in 1 patient. Fracture fixation included: plate-and-screws following arthroscopic reduction (1); screws (2) and threaded Steinmann pins (2); 2 fractures were debrided. Three patients had immediate (2) or delayed (1) lateral ulnar collateral ligament repair. RESULTS: Follow-up averaged 31.8 months. The range of motion averaged 9 degrees to 133 degrees in flexion-extension and 87 degrees/79 degrees in prono-supination. Mayo Elbow Performance scores were 100 in 5 of 6 patients and unavailable in 1 patient. Complications included asymptomatic heterotopic ossification (1) and delayed ulnar neuropathy (1). CONCLUSIONS: In our series of coronoid fractures treated with arthroscopic means, all patients had a functional, pain-free result. All patients returned to preoperative avocations and occupations. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17916471 TI - Surgical outcome of 2-stage management of multiple knee ligament injuries after knee dislocation. AB - PURPOSE: The purpose of this study was to determine the effectiveness of a new 2 stage surgical treatment for acute traumatic knee dislocation. METHODS: The study involved 15 knees in 14 patients treated between October 1997 and November 2001. The mean follow-up was 88.9 months (range, 35 to 110 months). In the first surgical stage, medial or lateral ligament complexes were repaired or reconstructed within 2 weeks of injury. In the second surgical stage, once full range of motion was obtained 3 to 6 months later, the anterior cruciate ligament (ACL) or posterior cruciate ligament (PCL) was reconstructed if significant laxity was present. The final outcomes were assessed based on stress radiographs, range of motion, Lysholm score, Tegner activity stage, and International Knee Documentation Committee rating. RESULTS: There were 10 cases of medial collateral ligament (MCL) tear and 8 cases of lateral collateral ligament (LCL) tear. All MCL and LCL injuries were either repaired or reconstructed. All cases had both ACL and PCL tears. After the first stage of MCL/LCL complex surgery, the second stage surgery involving ACL and PCL reconstruction was deemed necessary in 3 and 7 cases, respectively. Five cases did not require ACL or PCL reconstruction. On stress radiography at the last follow-up, MCL, LCL, ACL, and PCL instability was graded as 0 to 1 in 15, 14, 15, and 11 cases, respectively. PCL instability was graded as 2 in 4 cases. The mean postoperative Lysholm score was 87.6 points. CONCLUSIONS: The 2-stage surgical approach described resulted in good outcomes for acute knee dislocation patients in terms of range of motion and stability. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17916472 TI - Arthroscopic techniques for the fixation of a three-dimensional scaffold for autologous chondrocyte transplantation: structural properties in an in vitro model. AB - PURPOSE: The aim of the present study was to evaluate the structural properties of matrix-associated autologous chondrocyte implantation with multiple fixation techniques implanted in fresh porcine knees after they had undergone load to failure. METHODS: We evaluated the ultimate failure load, yield load, and stiffness of 3 different techniques for the fixation of a 2-mm thick polymer fleece: (1) fixation with biodegradable polylevolactide pins, (2) a transosseous anchoring technique, and (3) conventional suture fixation. Techniques 1 (pin) and 2 (transosseous anchoring) can be used arthroscopically. RESULTS: Maximum load and yield load were significantly higher in the group 1 (pin fixation) and group 2 (transosseous anchoring) compared to group 3 (conventional suture). Stiffness was significantly higher in group 1 than in groups 2 or 3. CONCLUSIONS: Our biomechanical data show that two fixation techniques (pin fixation and transosseous anchoring) have a higher ultimate load, yield load, and stiffness than the conventional suture technique at time point zero. However, these data can be interpreted only with the Bioceed-C matrix (BioTissue Technologies GmbH, Freiburg, Germany). CLINICAL RELEVANCE: Our biomechanical data show outstanding fixation strength with arthroscopic techniques that use Bioceed-C matrix scaffolds during autologous chondrocyte transplantation. Thus, arthroscopic fixation done with this biomaterial should benefit patients, which, in turn, should lead to further research on these arthroscopic techniques and this biomaterial. PMID- 17916473 TI - The reproducibility of radiographic measurement of lateral meniscus horn position. AB - PURPOSE: The objective of our investigation was to evaluate whether it is possible to locate and reproduce the tibial insertion areas of the anterior and posterior horns of the lateral meniscus on preoperative radiographs. METHODS: In 20 tibia heads, we prepared anterior and posterior horn insertions and marked their circumference with radiopaque steel balls of 1.6 mm in diameter. Standardized anteroposterior and lateral radiographs were made. On these radiographs, different landmarks were defined, their distances measured (tibial width and depth, distance from lateral tibia border to meniscus insertion midpoint, distance from anterior tibia border to meniscus insertion midpoint, distance from anterior and lateral tibia border to lateral intercondylar spine), and ratios determined. RESULTS: The anterior horn midpoint is located at 45.1% +/ 1.3% of tibial width and 41.9% +/- 3.2% of tibial depth, and the posterior horn midpoint is located at 49.8% +/- 1.9% of tibial width and 72.1% +/- 2.3% of tibial depth. The statistical analysis of these measures showed a precise and constant positioning of the lateral meniscus insertions on the tibia plateau. We also found constant topographic relations to the lateral intercondylar spine. CONCLUSIONS: Anterior and posterior lateral meniscus horn insertions can be determined on radiographs with a high precision and reproducibility. CLINICAL RELEVANCE: We have developed a technique for precise radiographic tibial horn determination in lateral meniscus transplantation. PMID- 17916474 TI - Evaluation of tendon graft fixation using alpha-BSM calcium phosphate cement. AB - PURPOSE: Many ligament reconstruction procedures require tendon healing within a surgically prepared bone tunnel. The interface between the graft and tunnel wall is the "weak link" in the initial phase of this process. By providing additional fixation and serving as a carrier for bone-derived growth factors, calcium phosphate cements may augment healing. The purpose of this study was to evaluate alpha-BSM calcium phosphate cement as the sole method of fixation for tendon and bone-tendon-bone grafts in an in vitro porcine model. METHODS: Ten tendon grafts and ten bone-tendon-bone grafts were fixed within porcine femoral and tibial bone tunnels by use of alpha-BSM calcium phosphate cement. The grafts were then loaded to failure. The ultimate pullout strength and the site of failure were recorded for each specimen. RESULTS: The bone-tendon-bone grafts achieved a mean pullout strength of 224.8 N (+/-79.9 N). Of the grafts, 6 failed at the cement-bone plug interface, 2 failed because of fracture of the bone plug, and 2 failed at the tendon-bone plug interface. The tendon grafts had a mean load to failure of 81.2 N (+/-23.7 N), and all 10 failed at the tendon-cement interface. The difference in fixation strength between these graft types was significant (P < .05). CONCLUSIONS: Though inadequate as the sole means of graft fixation, alpha-BSM calcium phosphate cement can help to augment bone plug fixation during the early phase of tendon-bone healing. In addition, alpha-BSM may provide added graft fixation and increased stiffness and act as a growth factor carrier for both tendon grafts and bone-tendon-bone grafts in ligament reconstruction surgery. CLINICAL RELEVANCE: This study evaluated the ability of alpha-BSM cement to augment tendon-bone healing because it may play a role as a growth factor carrier in future tendon healing models. PMID- 17916475 TI - Femoral bioabsorbable cross-pin fixation in anterior cruciate ligament reconstruction. AB - PURPOSE: The purpose of this study was to retrospectively evaluate clinical results after arthroscopic anterior cruciate ligament (ACL) reconstruction by use of hamstring autograft with femoral fixation via 2 bioabsorbable cross pins (Rigidfix system; DePuy Mitek, Raynham, MA). METHODS: We evaluated the results of 117 knees (117 patients) that had been treated with arthroscopic ACL reconstruction by use of hamstring autograft with femoral fixation via 2 bioabsorbable cross pins from September 2001 to November 2002. The mean follow-up period was 26.9 months (range, 25 to 32 months). Patients were evaluated by the Lachman test, pivot-shift test, KT-2000 arthrometer (MEDmetric, San Diego, CA) test, Lysholm score, and International Knee Documentation Committee grade at the preoperative and follow-up examinations. Of the knees, 74 were assessed by second look arthroscopy. RESULTS: The mean Lysholm score was 72.6 (range, 51 to 86) preoperatively and 93.7 (range, 71 to 99) postoperatively. On the basis of the final International Knee Documentation Committee grade, 75 knees were normal (A), 36 were nearly normal (B), 6 were abnormal (C), and none were severely abnormal (D). KT-2000 arthrometer analysis revealed that 112 knees (95.7%) were grade A or B, with a median laxity of 1.3 mm (range, 1 to 6 mm) at final follow-up. Of the 74 knees that were assessed by second-look arthroscopy, 52 had preserved good tension, 22 had some laxity, and none had graft failure or rupture. CONCLUSIONS: ACL reconstruction via 4-strand hamstring autograft with femoral fixation by use of 2 bioabsorbable cross pins eliminated anterior tibial translation in 93.1% of patients at a mean follow-up of 26.9 months. The bioabsorbable cross-pin femoral fixation method via hamstring tendon autograft can be effective, useful, and reproducible. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17916476 TI - Do anterior cruciate ligament allograft culture results correlate with clinical infections? AB - PURPOSE: In 1998, four cases of contaminated allografts for anterior cruciate ligament (ACL) reconstruction resulted in Clostridium infection, and a patient with Clostridium infection from a femoral condylar allograft died. It was subsequently published that implanting surgeons should culture ACL allografts so that action could be taken should highly pathogenic bacteria be encountered. The purpose of this study is to test the hypothesis that ACL allograft cultures correlate with clinical infections. METHODS: Since October 2003, a single surgeon performing ACL reconstruction prospectively cultured all allografts in the operating room before implantation. After culture, grafts were thawed in warm saline mixed with bacitracin. All patients received a single dose of preoperative antibiotics. Final culture results were obtained in all patients, and all patients were followed for a minimum of 90 days to evaluate for postoperative infection. The cost of cultures was determined by multiplying hospital charges by the hospital cost-to-charges ratio. RESULTS: Two hundred and ten cases were included. Ten allografts (4.8%) had positive culture results (6 coagulase negative Staphylococci, 1 alpha-Streptococcus-not-group-B, 1 Enterobacter, 1 Clostridium, and 1 polymicrobial [Klebsiella, Escherichia coli, and Enterococcus]). None of these patients had signs of infection; the three positive highly pathogenic bacteria (Enterobacter, Clostridium, and polymicrobial) graft recipients were treated with antibiotics. The others were observed. One patient with negative cultures developed Staphylococcus aureus infection. Mean culture cost was $127 (USD). CONCLUSIONS: Our results demonstrate that ACL allograft cultures do not correlate with clinical infections. LEVEL OF EVIDENCE: Level I, diagnostic study (testing of previously developed diagnostic criteria [culture]) in a series of consecutive patients (with universally applied reference gold standard [clinical evaluation for knee sepsis]). PMID- 17916477 TI - Reducing the "killer turn" in posterior cruciate ligament reconstruction by fixation level and smoothing the tibial aperture. AB - PURPOSE: The aim of this study was to evaluate the influence of the operative technique on reducing the so called "killer turn" at the tibial bone tunnel exit in posterior cruciate ligament (PCL) reconstruction. METHODS: We investigated the benefit of reducing the sharp edge of the tibial bone tunnel exit using a custom made rasp in combination with extracortical and aperture (interference screw) graft fixation. Grafts were fixed in a porcine tibia model. Ten reconstructions each (sharp and rounded edge) were cyclically loaded (2,000 times) between 50 and 150 N, and another 10 each were subjected to 2,000 cycles between 50 and 300 N force. The surviving PCL reconstructions were loaded to failure using a material testing machine. RESULTS: Being subjected to the sharp edge of the posterior tibia, 5 of 10 extracortical and 8 of 10 aperture fixed grafts survived cyclic loading between 50 and 150 N. All extracortical fixed and 8 of 10 aperture fixed grafts failed before 2,000 cycles when loaded between 50 and 300 N. Structural properties of grafts fixed with interference screw were statistically significant higher when compared to extracortical fixation. After rounding the sharp edge of the tunnel, all grafts survived cycles between 50 and 150 N and 6 out of 10 extracortical and 8 of 10 aperture fixed grafts survived 2,000 cycles between 50 and 300 N. CONCLUSIONS: The results of this study suggest that a rounded posterior aspect of the tunnel exit at the tibial tunnel exit leads to significant less graft damage when compared to the typical sharp edge of the bone tunnel exit ("killer turn"). Additionally, the results show that aperture fixation of soft tissue grafts in PCL reconstruction is superior when compared to an extracortical fixation site. CLINICAL RELEVANCE: Aperture fixation and a rounded tibial bone tunnel exit seem to be a reasonable alternative in PCL reconstruction. PMID- 17916478 TI - The risk of vascular injury to the femoral head when using the posterolateral arthroscopy portal: cadaveric investigation. AB - PURPOSE: The aim was to clarify the relationship between the posterolateral arthroscopy portal to the hip joint and the deep branch of the medial femoral circumflex artery (MFCA) and its terminal divisions. METHODS: In 7 fresh fixated human cadavers, the deep femoral arteries were injected with latex. In all specimens, the standard anterior, anterolateral, and posterolateral arthroscopy portals to the central compartment of the hip were established in a standardized procedure as done in our operating room. Subsequently, all specimens underwent surgical dissection to identify the tract of the posterolateral portal, the course of the deep branch of the MFCA, and its terminal divisions. The distance between the portal tract and the deep branch of the MFCA was measured at its shortest point. RESULTS: None of the specimens revealed injury to the deep branch of the MFCA and its terminal divisions; however, 1 specimen did not reveal filling of the MFCA, and did not allow for accurate measurement. For the remaining 6 specimens, the mean distance of the portal tract to the vessels at the closest point was 10.16 mm (standard deviation, 4.4 mm; range, 5 to 15 mm). In all specimens, the greater trochanter seemed to prevent closer contact between the vessel and the portal tract. CONCLUSIONS: As long as the shape of the posterior aspect of the greater trochanter has not been changed by previous surgery, the posterolateral portal to the hip joint can be performed safely with respect to the vital blood supply to the femoral head, because the MFCA is protected by the overhang of the posterior aspect of the greater trochanter, giving a minimum of 3 mm and an average 10 mm of clearance. CLINICAL RELEVANCE: At the level of the piriformis muscle, the posterolateral portal has a minimum of 3 mm and an average of 10 mm of clearance to the deep branch of the MFCA. As long as the shape of the posterior aspect of the greater trochanter has not been changed, and the skin penetration remains close to this bony margin with direction of the portal toward the joint space, this portal can be performed safely with respect to the vital blood supply to the femoral head. PMID- 17916479 TI - Modified posterior portals for hindfoot arthroscopy. AB - PURPOSE: The purpose of this study was to determine the course and safe distances achieved with modified coaxial portals for hindfoot arthroscopy and report the clinical results. METHODS: We used 30 embalmed cadaveric ankle specimens and 10 fresh-frozen ankle specimens for anatomic measurements and trial operations. The posteromedial portal via the posterior tibial tendon sheath was first established. The posterolateral portal was subsequently created immediately behind the posterior border of the lateral malleolus and anterior to the peroneal tendons via an inside-out technique. The coaxial portals were finally established with cannulas left in place. In the clinical series, posterior ankle arthroscopy was performed on 18 ankles in 15 patients. All patients were evaluated for any complications with a mean follow-up of 38 months. RESULTS: The posterior tibial nerve, posterior tibial artery, and peroneal artery were located a mean distance of 8.7 mm, 10.1 mm, and 12.9 mm, respectively, from the near edge of the Kirschner wire as a reference to the coaxial portals. The sural nerve and lesser saphenous vein were at greater distances of 27.6 mm and 28.3 mm, respectively. The mean West Point score at the time of the latest follow-up was 91.5 points (range, 76 to 100 points), and there were 9 excellent results, 3 good results, and 1 fair result. No patients showed any complications related to the modified coaxial portals. CONCLUSIONS: The modified coaxial portals seemed to have large distances to the neurovascular structures in our anatomic study. Clinically, this technique was safe, effective, and reproducible. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17916480 TI - The use of anchors in shoulder surgery: a shift from metallic to bioabsorbable anchors. AB - With major advances in arthroscopy, suture anchors became the primary devices used to assist in fixing soft tissues to bone. Metallic anchors were first produced and used in soft tissue fixation around the shoulder. However, their use resulted in some reported complications, including articular surface damage from migrating implants and distortion and artifact production in postoperative magnetic resonance imaging. Bioabsorbable anchors were developed to avoid these problems. Their newer versions were proven to have pulling-out strength equal to that of metallic anchors, with a reported lower complication rate. This had led to a major shift away from metallic anchors toward bioabsorbable anchors. PMID- 17916481 TI - Arthroscopic treatment of unresolved Osgood-Schlatter lesions. AB - Osgood-Schlatter disease is a self-limiting condition in most cases. Those with unresolved pain after conservative treatment can obtain relief with surgical debridement of the mobile ossicles and tibial tuberosity. We present an arthroscopic technique for debridement. The location of the inferomedial and lateral parapatella tendon portals can be raised slightly to allow improved instrumentation and visualization in the anterior interval. An anterior interval release is performed with the mechanical shaver and radiofrequency ablation device. Care is taken to visualize the meniscal anterior horns and intermeniscal ligament. By staying anterior to these structures, debridement can be performed aggressively onto the anterior tibial slope. The bony lesions are shelled out from their soft-tissue attachments. Small and loose fragments are removed with a pituitary ronguer, whereas larger lesions are removed with an arthroscopic burr. Working deep along the anterior tibial slope is facilitated by extending the knee and taking tension off the patellar tendon. Postoperatively, patients are allowed full weight bearing and unrestricted range of motion. The advantages of this technique include the avoidance of the patellar tendon longitudinal split required for open procedures and the ability to address concomitant intra articular pathology. PMID- 17916482 TI - Arthroscopic identification of the anterior cruciate ligament posterolateral bundle: the figure-of-four position. AB - Anatomic double-bundle reconstruction in anterior cruciate ligament (ACL) tears has been developed during the last few years. Although anteromedial (AM) bundle reconstruction is routinely performed, finding the femoral attachment of the posterolateral (PL) bundle remains a problem. We describe how a classic arthroscopic position, the figure-of-4 position, allows the PL bundle to be easily recognized. During flexion, the femoral attachment of the PL bundle describes an arc around the AM femoral attachment. The femoral attachment of the AM bundle is the center of rotation of the ACL, which explains the isometric behavior of this bundle. After 45 degrees of flexion, the PL femoral attachment becomes anterior to the AM femoral attachment. The AM bundle is tight during flexion, whereas the PL bundle is under tension when the knee is extended and becomes lax with knee flexion. At 90 degrees of flexion, the PL bundle is difficult to identify because it is lax; only its femoral insertion lies anterior to the AM bundle close to the articular cartilage of the lateral condyle. The use of an additional tibial varus torque and internal rotation (i.e., the figure-of-4 position) tightens the PL bundle and enhances the visualization of its insertion, allowing for easy identification of this bundle. PMID- 17916483 TI - All-inside suture technique for Palmer class 1B triangular fibrocartilage repair. AB - After completion of the diagnostic procedure, the ulnar-sided tear is identified and tear debridement is performed. A 1.5-cm incision is made to isolate the sensory branch of the ulnar nerve. This is made in the region of the 6U portal. The arthroscope is left in the 3-4 portal, and the slotted needle is inserted through the capsule wall, avoiding the isolated sensory ulnar branch, penetrating the ulnar-sided tear of the triangular fibrocartilage complex. A No. 2 PDS monofilament suture (Ethicon, Somerville, NJ) is inserted into the needle and visualized in the joint. The end of the suture is pulled through the 6R portal. The slotted needle is then removed and detached from the suture. The same needle is inserted through the tear about 0.3 cm from the point where the needle exited on the previous suture. The other end of the suture is inserted into the needle, and once visualized within the joint, it is withdrawn again through the 6R portal. In this way, we create an extra-articular loop with both ends of the same suture passing through the tear by way of the 6R portal. A knot is then tied and fixed inside the joint on the ulnar side of the triangular fibrocartilage complex. By use of the same technique, 2 or 3 more stitches can be placed, improving tear security. PMID- 17916484 TI - Arthroscopic-assisted reduction and percutaneous external fixation of lateral condyle fractures of the humerus. AB - Lateral condyle fractures in children are the second most common fracture about the elbow. Anatomic reduction of intra-articular displacement is considered a priority. Most experts agree that the treatment of choice of mildly displaced (>2 mm) lateral condyle fractures is open reduction-internal fixation or intraoperative arthrography followed by closed reduction and percutaneous pinning. In this report we described an alternative approach using closed arthroscopic-assisted reduction. Arthroscopy was performed by use of a 30 degree, 4.5-mm arthroscope through a proximal anteromedial portal with no pump, and a proximal anterolateral portal was created for instrumentation. The fracture line was easily visualized after blood, fibrin, and loose articular debris were irrigated from the joint. The fracture was manually manipulated and reduced by use of a periosteal elevator placed through the proximal anterolateral portal into the fracture and via rotation until there was no step in the articular cartilage. Two lateral Kirschner wires were placed percutaneously, which result in firm fixation of the fracture. We think that the advantages of arthroscopy are further superior to intraoperative arthrography because arthroscopy not only allows for better anatomic reduction and visualization with minimal surgical trauma and less radiation time but also provides a valid diagnostic and treatment alternative for associated injuries. PMID- 17916485 TI - Minimally invasive coracoclavicular ligament augmentation with a flip button/polydioxanone repair for treatment of total acromioclavicular joint dislocation. AB - Treatment of complete acromioclavicular joint disruption remains controversial and ranges from rehabilitation to extensive surgical reconstruction. However, high-grade injuries (type IV, V, and VI) are typically treated surgically. Most reconstruction techniques addressing these injuries selectively focus on coracoclavicular ligament augmentation because it has been shown to be the primary stabilizer of the acromioclavicular joint. The conventional coracoclavicular polydioxanone (PDS) loop, which is widely performed, has been detected to have some pivotal disadvantages, including anterior subluxation of the clavicle, extensive preparation of the coracoid, and bony avulsion of the clavicle as a result of rotational clavicle movement. Therefore we present an augmentation technique that reduces these complications by replicating the orientation of the native coracoclavicular ligament complex and providing a minimally invasive subcoracoid and clavicular fixation of a double PDS loop by use of 2 flip buttons, typically used for extracortical anterior cruciate ligament graft fixation. The key step of the procedure includes the anatomic, secure, and stable placement of the double PDS cerclage under the coracoid base transferring a flip button through a coracoid bone tunnel. Our clinical experience shows that the presented technique is easy to perform and has a comparable invasiveness to recently presented arthroscopic techniques. PMID- 17916486 TI - Arthroscopic double-row suture anchor fixation of minimally displaced greater tuberosity fractures. AB - In cases of displaced greater tuberosity fractures, treatments by arthroscopic assisted reduction and percutaneous screw fixation have been reported. However, in cases in which there is a comminuted fracture or a minimally displaced fracture combined with concomitant lesions such as rotator cuff tear or labral pathology, it is difficult to reduce the fracture and to treat other pathologies by use of a percutaneous screw. Recently, many surgeons have used the double-row repair method in rotator cuff repair, which provides a tendon-bone interface better suited for biologic healing and restoring normal anatomy. In accordance with this method, we used the arthroscopic technique of double-row suture anchor fixation for a minimally displaced greater tuberosity fracture without additional incision. Initially, debridement was performed on the fracture surface by use of a shaver, and the medial-row anchor was inserted through the anterior portal or the intact cuff. Two lateral-row anchors were inserted just anterior and posterior to the lower margin of the fractured fragment under C-arm guidance. The medial-row sutures and lateral-row sutures were then placed. Arthroscopic double row suture anchor fixation of a displaced greater tuberosity fracture restores the original footprint of the rotator cuff and normal tendon-bone interface of the displaced greater tuberosity fracture. PMID- 17916487 TI - Arthroscopic release of the superior transverse ligament and SLAP refixation in a case of suprascapular nerve entrapment. AB - We report a case of suprascapular nerve entrapment at the suprascapular notch combined with a type II SLAP lesion resulting in supraspinatus and infraspinatus muscle weakness and shoulder pain in a 27-year-old female professional handball player. The magnetic resonance imaging scan showed significant atrophy of the supraspinatus and infraspinatus muscles. Electromyography revealed an isolated proximal lesion of the suprascapular nerve. The patient was treated by an arthroscopic release of the superior transverse ligament and repair of the type II SLAP lesion. Follow-up evaluations were performed 6 weeks, 3 months, and 6 months postoperatively. The Constant score improved from 51 to 84 points. Electromyography studies 3 and 6 months after surgery showed significant improvement with normal reinnervation of the supraspinatus and infraspinatus muscles. To our knowledge, this is the first report of proximal suprascapular nerve entrapment with coincidence of a SLAP lesion that was treated arthroscopically. On the basis of this case, we found that arthroscopic release of the superior transverse ligament is an effective procedure for decompression of the suprascapular nerve. Although it is a technically demanding procedure, the arthroscopic approach has the advantage of detecting concomitant lesions such as SLAP lesions. PMID- 17916488 TI - Superior Bennett lesion: a bone fragment at the posterosuperior glenoid rim in 5 athletes. AB - In throwing athletes, a Bennett lesion is a bony spur that usually forms at the posteroinferior glenoid rim and sometimes becomes painful. We encountered superior Bennett lesions in the shoulders of 5 athletes, which were detected at the posterosuperior glenoid rim as a separate bone fragment. In all 5 athletes this lesion appeared to be the main cause of shoulder pain during throwing or overhead activity. The lesions were easily visualized on plain radiographs (axial view, scapular Y view, and 45 degrees craniocaudal view) or computed tomography scans. Tenderness was detected over the posterior aspect of the glenohumeral joint, the pain provocation test specific for SLAP lesions was positive, and posterior shoulder pain was elicited by forced external rotation at 90 degrees of abduction in all 5 cases. The pain was diminished or decreased by injection of local anesthetic around the lesion. At arthroscopy, detachment of the posterosuperior labrum and posterior capsular tightness were detected in all 5 cases. Through the detached portion of the posterosuperior labrum, we could easily reach an unstable mobile bone fragment, which was resected arthroscopically. Although the pathologic mechanism is unclear, the mobility of the bone fragment appeared to be related to the occurrence of symptoms. PMID- 17916489 TI - Bilateral recurrent patellar dislocation in a patient with isolated patella aplasia-hypoplasia. AB - Patella aplasia-hypoplasia is a rare condition characterized by the congenital absence or marked reduction of the patellar bone. It is well known that patella aplasia-hypoplasia occurs in nail-patella syndrome, small patella syndrome, and several other syndromes. However, isolated patella aplasia-hypoplasia without associated clinical or radiologic anomalies, first described by Kutz in 1949, is an extremely rare condition. This report describes 12 years' follow-up after medial patellofemoral ligament reconstruction surgery performed for bilateral recurrent patellar dislocation in a 29-year-old patient with isolated patella aplasia-hypoplasia. The patient had no radiographic findings of elbow abnormalities, exostoses of the ilia, or abnormal ossification of the ischiopubic junction. The right patella was severely subluxated, and the left patella was mildly subluxated. The preoperative Kujala score was 28 points for the right patella and 23 points for the left. Medial patellofemoral ligament reconstruction, wide lateral release, and patellaplasty with multiple drilling were done in July 1993 for the left knee and in August 1993 for the right. At 12 years' follow-up, the Kujala score was 68 points for the right patella and 73 points for the left, and the patient was satisfied with the result. Magnetic resonance images showed both patellae without subluxation and bulky regenerated ligaments. PMID- 17916490 TI - Quantitative information from ultrasound evaluation of articular cartilage should be interpreted with care. PMID- 17916492 TI - Single-portal SLAP lesion repair. PMID- 17916494 TI - Current and emerging trends in the treatment of early-stage colorectal cancer: importance of a multidisciplinary approach. PMID- 17916496 TI - The sentinel node biopsy under local anesthesia in breast cancer: advantages and problems, how the technique influenced the activity of a breast surgery department; update from the European Institute of Oncology with more than 1000 cases. AB - Sentinel lymph node biopsy (SLNB) is a staging technique with a significant impact on patients' quality of life: the oncological effectiveness in a large number of patients affected by breast carcinoma has been already demonstrated, and the clinical research is now focusing on new indication for the biopsy and widespread adoption of the technique. At the European Institute of Oncology we are applying SLNB under local anesthesia: our aim is to improve the management of the disease with low costs for the structure and patients, and to improve patients' acceptance of breast cancer treatments. We are now discussing the impact of the SLNB under local anesthesia on the activity of a breast surgery department. We also present an update of our experience. PMID- 17916497 TI - REPAT, a new family of proteins induced by bacterial toxins and baculovirus infection in Spodoptera exigua. AB - Insect larvae spend most of their time eating and the digestive tract is the most crucial barrier for the entrance of many pathogens. In our study, suppression subtractive hybridization (SSH) was used to compare Spodoptera exigua midgut gene expression between larvae exposed to the Bacillus thuringiensis Cry1Ca toxin and non-exposed insects. Based on the SSH results, full cDNA sequences coding for four homologous proteins were obtained. Quantitative and semi-quantitative RT-PCR showed the increased expression of the genes coding for these proteins after exposure to different B. thuringiensis toxins as well as after infection with baculovirus. The proteins were named REPAT after their increased expression in Response to Pathogen. REPAT1, a member of this family, was recombinantly expressed using the baculovirus expression system, revealing the glycosylated nature of the protein. Recombinant baculoviruses expressing REPAT1 were used to infect larvae from S. exigua, showing that expression of REPAT1 was reducing the virulence of baculovirus to the infected larvae. Together, these results suggest a role for REPAT1 in mitigating pathological effects. PMID- 17916498 TI - Host plant-dependent metabolism of 4-hydroxybenzylglucosinolate in Pieris rapae: substrate specificity and effects of genetic modification and plant nitrile hydratase. AB - After ingestion of transgenic Arabidopsis thaliana CYP79A1 containing sinalbin (4 hydroxybenzylglucosinolate) due to genetic modification, only one major sinalbin derived sulphate ester (the sulphate ester of 4-hydroxyphenylacetonitrile) was excreted by Pieris rapae caterpillars (corresponding to 69mol% of ingested sinalbin). An additional sulphate ester (the sulphate ester of 4 hydroxyphenylacetamide) was excreted when the caterpillars were reared on two plant species (Sinapis alba and Sinapis arvensis) that contained sinalbin naturally. Artificial addition of sinalbin to S. arvensis leaves resulted in increased levels of the sulphated amide, and an enzymatic activity (nitrile hydratase) explaining the formation of the sulphated amide from sinalbin was detected in both Sinapis species, but not in A. thaliana. In agreement with the suggested minor metabolic pathway, the caterpillars were able to sulphate 4 hydroxyphenylacetamide offered as part of an artificial diet. In fact, phenol and seven para-substituted phenol derivatives with substituents of moderate size were sulphated and excreted, but all tested phenols devoid of a nitrile functional group were less efficiently sulphated than the primary sinalbin detoxification product, 4-hydroxyphenylacetonitrile. This suggests that the specificity of the sulphation step involved in sinalbin metabolism may be adapted to nitriles formed as metabolites of phenolic glucosinolates. On the contrary, there was no specificity for products (4-hydroxybenzylascorbigen and 4-hydroxybenzylalcohol) derived from the semistable isothiocyanate produced from sinalbin in the absence of nitrile specifier protein. PMID- 17916499 TI - Characterization of novel esterases in insecticide-resistant mosquitoes. AB - In the mosquito Culex pipiens complex (Diptera: Culicidae), the amplification of carboxylesterase genes is an important mechanism providing resistance to organophosphate insecticides. Various amplified alleles at the Ester locus have been identified over the world. In this study, two newly detected Ester alleles, Ester(B10) and Ester(11) (including associated Ester(A11) and Ester(B11)), coding for esterases B10 and A11-B11, respectively, are characterized qualitatively and quantitatively. A high molecular identity is observed both at the nucleotide level and at the deduced amino acid level among the known Ester alleles. Real time quantitative PCR results suggest 2.5-fold amplification of the Ester(B10) allele, 36.5-fold amplification of the Ester(A11) allele, and 19.1-fold amplification of the Ester(B11) allele. The ca. 2-fold difference in amplification level between Ester(A11) and Ester(B11) may indicate a new model for the esterase amplification. Bioassays show that these two resistant Ester alleles only can confer moderate or low resistance to the tested organophosphate insecticides. PMID- 17916500 TI - Functional expression and characterisation of a gut facilitative glucose transporter, NlHT1, from the phloem-feeding insect Nilaparvata lugens (rice brown planthopper). AB - Phloem-sap feeding Hemipteran insects have access to a sucrose-rich diet but are dependent on sucrose hydrolysis and hexose transport for carbon nutrition. A cDNA library from Nilaparvata lugens (rice brown planthopper) was screened for clones encoding potential transmembrane transporters. A selected cDNA, NlHT1, encodes a 53kDa polypeptide with sequence similarity to facilitative hexose transporters of eukaryotes and prokaryotes, including GLUT1, the human erythrocyte hexose transporter. NlHT1 was expressed as a recombinant protein in the methylotropic yeast Pichia pastoris, and was identified in a membrane fraction isolated from transformed yeast cells. Transport experiments using membrane vesicles containing NlHT1 showed that the protein is a saturable, sodium independent transporter, with a relatively low affinity for glucose (K(m) 3.0mM), which can be inhibited by cytochalasin B. Competition experiments with fructose demonstrate NlHT1 is glucose specific. In situ localisation studies revealed that NlHT1 mRNA is expressed in N. lugens gut tissue, mainly in midgut regions, and that expression is absent in hindgut and Malpighian tubules. NlHT1 is therefore likely to play an important role in glucose transport from the gut, and in carbon nutrition in vivo. This is the first report of a facilitative glucose transporter from a phloem-feeding insect pest. PMID- 17916501 TI - A proteomic approach to the identification of salivary proteins from the argasid ticks Ornithodoros moubata and Ornithodoros erraticus. AB - The saliva of ticks contains anti-haemostatic, anti-inflammatory and immunomodulatory molecules that allow these parasites to obtain a blood meal from the host and help tick-borne pathogens to infect the vertebrate host more efficiently. This makes the salivary molecules attractive targets to control ticks and tick-borne pathogens. Although Ornithodoros moubata and O. erraticus are important argasid ticks that transmit severe diseases, to date only a few of their salivary proteins have been identified. Here we report our initial studies using proteomic approaches to characterize the protein profiles of salivary gland extracts (SGE) from these two argasids. The present work describes the proteome of the SGEs of both tick species, their antigenic spots, and the identification of several of their proteins. The whole number of identifications was low despite the good general quality of the peptide mass maps obtained. In the O. moubata SGE, 18 isoforms of a protein similar to O. savignyi TSGP1 were identified. In the O. erraticus SGE we identified 6 novel proteins similar to unknown secreted protein DS-1 precursor, NADPH dehydrogenase subunit 5, proteasome alpha subunit, ATP synthase F0 subunit 6, lipocalin and alpha tubulin. Finally, the current drawbacks of proteomics when applied to the identification of acarine peptides and proteins are discussed. PMID- 17916502 TI - Upregulation of a desaturase is associated with the enhancement of cold hardiness in the onion maggot, Delia antiqua. AB - Cold-acclimated non-diapause pupae, and summer- and winter-diapause pupae of the onion maggot, Delia antiqua (Diptera: Anthomyiidae), show marked cold hardiness as compared with intact non-diapause pupae. Homeoviscous adaptation of cellular membranes is crucial to enhance the cold hardiness of organisms, and Delta9-acyl CoA desaturases have been assumed, albeit without experimental evidence in insects, to play a key role in the adaptation. We cloned the cDNA of a desaturase gene (Dadesat) from D. antiqua, which is most likely to encode Delta9-acyl-CoA desaturase. Expression of Dadesat mRNA in the brain, midgut, and Malpighian tubules of cold-acclimated and diapause pupae was upregulated 2-10 fold, correlating well with the increase in cold hardiness. In the pupae with enhanced cold hardiness, palmitoleic and oleic acids, the presumed products of Dadesat, in the phospholipids were significantly increased. These findings suggest that the increase in the expression of Dadesat contributes to enhanced cold hardiness in D. antiqua through the production of these unsaturated fatty acids. PMID- 17916503 TI - Genetic sexing through the use of Y-linked transgenes. AB - Sterile insect technique (SIT)-based pest control programs rely on the mass release of sterile insects to reduce the wild target population. In many cases, it is desirable to release only males. Sterile females may cause damage, e.g., disease transmission by mosquitoes or crop damage via oviposition by the Mediterranean fruit fly (Medfly). Also, sterile females may decrease the effectiveness of released males by distracting them from seeking out wild females. To eliminate females from the release population, a suitable sexual dimorphism is required. For several pest species, genetic sexing strains have been constructed in which such a dimorphism has been induced by genetics. Classical strains were based on the translocation to the Y chromosome of a selectable marker, which is therefore expressed only in males. Recently, several prototype strains have been constructed using sex-specific expression of markers or conditional lethal genes from autosomal insertions of transgenes. Here, we describe a novel genetic sexing strategy based on the use of Y-linked transgenes expressing fluorescent proteins. We demonstrate the feasibility of this strategy in a major pest species, Ceratitis capitata (Wiedemann), and discuss the advantages and disadvantages relative to other genetic sexing methods and potential applicability to other species. PMID- 17916504 TI - Proteomic profiling of a parasitic wasp exposed to constant and fluctuating cold exposure. AB - When insects are exposed to fluctuating thermal regimes (FTRs) (i.e., cold exposure alternating with periodic short pulses to high temperature), in contrast to constant low temperature (CLT), mortality due to accumulation of chill injuries is markedly reduced. To investigate the physiological processes behind the positive impact of FTR, based on a holistic approach, two-dimensional electrophoresis (2-DE) analysis were performed with the parasitic wasp Aphidius colemani. Parasitoid proteomes revealed 369 well-distinguishable protein spots, where the overall response to cold exposure was clearly specific to treatments (CLT versus FTR). The reduced mortality under FTR was associated with up regulation of several proteins playing key roles in energy metabolism (glycolysis, TCA cycle, synthesis and conversion of ATP), protein chaperoning (Hsp70/Hsp90), and protein degradation (proteasome). Our results also support the idea that cytoskeleton components, particularly actin arrangement, could play a role in the higher survival rates of insects under FTR. PMID- 17916505 TI - Intimate roles for cyanogenic glucosides in the life cycle of Zygaena filipendulae (Lepidoptera, Zygaenidae). AB - Zygaena larvae sequester the cyanogenic glucosides (CNglcs) linamarin and lotaustralin from their food plants (Fabaceae) and also de novo biosynthesize these compounds. In Zygaenidae, CNglcs serve as defence compounds during the entire life cycle, and their content and ratio are tightly regulated. We demonstrate that Z. filipendulae males transfer a nuptial gift of CNglcs to females during mating, and that females prefer males with a higher content of CNglcs for mating. Average HCN emission from female imagines is 19 times higher than from males, suggesting that plumes of HCN emitted from the perching female may serve to attract flying males. Analysis of the linamarin and lotaustralin content and ratio within different tissues in Z. filipendulae larvae shows that integument and haemolymph constitute the main sites of CNglc deposition. The data suggest that CNglcs may serve an additional role as storage compounds of reduced nitrogen that is mobilized during the transition of the last instar larva to imago, most likely to provide nitrogen for chitin synthesis. At least one of the enzymes responsible for de novo biosynthesis of CNglcs in Z. filipendulae is located in the integument. In conclusion, CNglcs play many important and different roles during the entire life cycle of Z. filipendulae in addition to defence. PMID- 17916506 TI - Cloning, expression and characterization of a dipteran farnesyl diphosphate synthase. AB - Farnesyl diphosphate synthase (FPPS) of the dipteran Drosophila melanogaster has been cloned and its catalytic properties have been assessed. Analysis of the D. melanogaster genome and of ESTs indicates that FPPS is a single copy gene that produces two transcripts, which differ only by 5' extension. The cDNA of shorter and longer D. melanogaster FPPSs (DmFPPS-1a and DmFPPS-1b, respectively) were each subcloned into pET28a and expressed as an N-His6 fusion protein in BL21 E. coli cells. The DmFPPSs similarly catalyzed the coupling of the allylic substrates, GPP and DMAPP, with IPP, producing FPP as product. The longer protein was further characterized. The enzyme required divalent metal for activity, and was activated by 0.1% Triton X-100. Higher detergent concentration and the addition of glycerol, conditions that activate certain insect FPPSs, inhibited prenyl coupling by DmFPPS-1b. Although DmFPPS-1b does not efficiently couple homologous GPP compounds, homodimethylallyl diphosphate (HDMAPP), which is precursor to all homologous JH structures, was a reactive substrate. PMID- 17916507 TI - Biochemical properties of the major proteins from Rhodnius prolixus eggshell. AB - Two proteins from the eggshell of Rhodnius prolixus were isolated, characterized and named Rp30 and Rp45 according to their molecular masses. Purified proteins were used to obtain specific antiserum which was later used for immunolocalization. The antiserum against Rp30 and Rp45 detected their presence inside the follicle cells, their secretion and their association with oocyte microvilli. Both proteins are expressed during the final stage of vitellogenesis, preserved during embryogenesis and discarded together with the eggshell. The amino terminals were sequenced and both proteins were further cloned using degenerated primers. The amino acid sequences appear to have a tripartite arrangement with a highly conserved central domain which presents a repetitive motif of valine-proline-valine (VPV) at intervals of 15 amino acid residues. Their amino acid sequence showed no similarity to any known eggshell protein. The expression of these proteins was also investigated; the results demonstrated that this occurred strictly in choriogenic follicles. Antifungal activity against Aspergillus niger was found to be associated with Rp45 but not with Rp30. A. niger exposed to Rp45 protein induced growth inhibition and several morphological changes such as large vacuoles, swollen mitochondria, multi-lamellar structures and a disorganized cell wall as demonstrated by electron microscopy analysis. PMID- 17916508 TI - Alternative splicing generates multiple transcripts of the inhibitor of apoptosis protein 1 in Aedes and Culex spp. mosquitoes. AB - We determined the sequences of cDNA encoding Inhibitor of Apoptosis Protein 1 (IAP1) homologues from Aedes triseriatus, Aedes albopictus, Aedes aegypti, Culex pipiens and Culex tarsalis. The cDNAs encode translation products that share > or = 84% sequence similarity. The IAP1 mRNA of each mosquito species exists as 3-5 distinct variants due to the presence of heterogeneous sequences at the distal end of their 5'UTRs. Partial genomic sequencing upstream of the 5' end of the Ae. triseriatus IAP1 gene, and analysis of the Ae. aegypti genomic sequence, suggest that these mRNA variants are generated by alternative splicing. Each IAP1 mRNA variant from Ae. triseriatus and Cx. pipiens was detected by RT-PCR in all mosquito life-stages and adult tissues examined, and the relative concentration of each Ae. triseriatus IAP mRNA variant in various tissues was determined. PMID- 17916510 TI - Brennan phenotype and its downstream expression. PMID- 17916509 TI - Identification and characterization of a novel sericin gene expressed in the anterior middle silk gland of the silkworm Bombyx mori. AB - Sericin is a group of proteins expressed in the middle silk gland that covers the surface of fibroin in the cocoon filament of Bombyx mori. Sericin consists of several serine-rich proteins with different molecular masses. Sericin A is one of the proteins and is produced in the anterior portion of the middle silk gland. To identify the gene coding for the protein, we determined the primary structures of its partial peptides, and the gene was searched using the silkworm genomic databases. Three contigs containing the corresponding nucleotide sequences were identified and categorized as one group. The gene structure covering the 5' flanking and the 3' end was determined by PCR fragments from genomic DNA, RT-PCR, and 5' and 3' RACE. The amino acid sequence deduced from the nucleotide sequence mainly consists of two serine-rich regions of 86-amino acid motif and 8-amino acid repeated sequence. The expression of the gene is limited to the anterior and middle parts of the middle silk gland. In addition, because the sericin gene appeared different from the sericin 1 and 2 genes reported earlier, we designated the newly discovered gene as sericin 3. PMID- 17916512 TI - Brennan-isms and the process of mentoring. PMID- 17916513 TI - An interim analysis from Dr Brennan's investment portfolio. PMID- 17916514 TI - How can we inspire residents to become surgical oncologists? PMID- 17916515 TI - Does intraoperative tumor cell dissemination matter? PMID- 17916516 TI - Role of surgery in Zollinger-Ellison syndrome. PMID- 17916517 TI - Minimally invasive parathyroidectomy and preoperative MIBI scans: correlation of gland weight and preoperative PTH. AB - BACKGROUND: Minimally invasive parathyroidectomy (MIP) has become increasingly popular for the treatment of patients with primary hyperparathyroidism (pHPT), and the specific techniques used vary from surgeon to surgeon. This strategy depends on preoperative localization tests, of which the MIBI scan is the most commonly used. This study details one surgeon's MIP experience and examines factors that correlate with the results of sestamibi (MIBI) scans. STUDY DESIGN: A retrospective review of parathyroidectomies performed between 1996-2006 was performed to create a database including laboratory and imaging results, symptoms, length of stay, and complications. MIBI scans were classified as correct when they showed one area of uptake at the site of a single adenoma (SA) found at exploration. Correlation between patient factors and MIBI scan results were assessed with Spearman correlation and Wilcoxon rank-sum tests. RESULTS: Of 263 patients having parathyroidectomy for pHPT, 205 had SA, 40 double adenomas, 15 hyperplasia, and 3 negative explorations. Normocalcemia was achieved in 98% of patients at 1 year and 95% by intraoperative parathyroid hormone (PTH) criteria. Factors that were significantly correlated with MIBI scan results in SA patients were preoperative PTH (p = 0.0025) and adenoma weight (p < 0.0001). The median PTH in those with correct scans was 137 versus 101 pg/mL in those with incorrect scans, and the median adenoma weights were 920 and 280 mg, respectively. CONCLUSIONS: The MIBI scan is a good localization test that made unilateral exploration and MIP possible in 76% of the cases. MIBI scans are more likely to be correct with higher preoperative PTH and larger adenomas. PMID- 17916519 TI - Adenocarcinoma of the esophagogastric junction: competition between Barrett and gastric cancer. PMID- 17916518 TI - New directions in the treatment of thyroid cancer. PMID- 17916520 TI - Adjuvant therapy for gastric cancer. PMID- 17916521 TI - A three and one-half decade nutritional and metabolic iliad. PMID- 17916522 TI - A new model of nutrition influenced inflammatory risk. PMID- 17916523 TI - Herpes viral oncolysis: a novel cancer therapy. PMID- 17916524 TI - Pancreatic adenocarcinoma: putting a hump in survival. PMID- 17916525 TI - Surgical treatment of pancreatic cancer. PMID- 17916526 TI - Pancreatic cancer--progress made. PMID- 17916527 TI - Sarcoma nomogram: validation and a model to evaluate impact of therapy. PMID- 17916528 TI - Surgical leaders and educators for the 21st century. PMID- 17916529 TI - "The little red book"--sayings of chairman Brennan. PMID- 17916531 TI - Global child health: burden of disease, achievements, and future challenges. PMID- 17916533 TI - Unusual cause of abdominal pain: Peutz-Jeghers syndrome. PMID- 17916534 TI - Gastrointestinal [corrected] stromal tumor of the ampulla of vater mimicking a duodenal ulcer. PMID- 17916535 TI - Meckel's diverticulum diagnosed by double-balloon enteroscopy. PMID- 17916536 TI - Meckel's diverticulum with recurrent bleeding diagnosed by double-balloon enteroscopy. PMID- 17916537 TI - Meckel's diverticulum on small-bowel follow-through series and computerized tomography scan in a 7-year-old boy with anemia. PMID- 17916541 TI - A 51-year-old with irritable bowel syndrome: test or treat for bacterial overgrowth? PMID- 17916540 TI - The narcotic bowel syndrome: clinical features, pathophysiology, and management. AB - Narcotic bowel syndrome (NBS) is a subset of opioid bowel dysfunction that is characterized by chronic or frequently recurring abdominal pain that worsens with continued or escalating dosages of narcotics. This syndrome is underrecognized and may be becoming more prevalent. In the United States this may be the result of increases in using narcotics for chronic nonmalignant painful disorders, and the development of maladaptive therapeutic interactions around its use. NBS can occur in patients with no prior gastrointestinal disorder who receive high dosages of narcotics after surgery or acute painful problems, and among patients with functional gastrointestinal disorders or other chronic gastrointestinal diseases who are managed by physicians who are unaware of the hyperalgesic effects of chronic opioids. The evidence for the enhanced pain perception is based on the following: (1) activation of excitatory antianalgesic pathways within a bimodal opioid regulation system, (2) descending facilitation of pain at the rostral ventral medulla and pain facilitation via dynorphin and cholecystokinin activation, and (3) glial cell activation that produces morphine tolerance and enhances opioid-induced pain. Treatment involves early recognition of the syndrome, an effective physician-patient relationship, graded withdrawal of the narcotic according to a specified withdrawal program, and the institution of medications to reduce withdrawal effects. PMID- 17916542 TI - Stiffness and impedance: the new liver biomarkers. PMID- 17916543 TI - An eosinophil hypothesis for functional dyspepsia. PMID- 17916544 TI - Angiotensin-converting enzyme inhibitors and risk of esophageal and gastric cancer: a nested case-control study. AB - BACKGROUND & AIMS: There are contradictory results regarding the association between angiotensin-converting enzyme (ACE) inhibitors and cancer. We aimed to investigate whether ACE inhibitors protect against esophageal and gastric cancer. METHODS: We conducted a population-based case-control study nested within the General Practitioners' Research Database in the United Kingdom. All individuals in the General Practitioners' Research Database aged 40-84 years between 1994 and 2001 were followed up until detection of an esophageal or gastric cancer (cases), other cancer, age of 85 years, death, or end of study period. RESULTS: Among 4.34 million person-years, 909 cases of esophageal and 1023 cases of gastric cancer were identified, and 10,000 matched controls were selected at random. Adjustments included smoking, body mass index, concurrent medication, and gastrointestinal disorders. Current use of ACE inhibitors decreased the risk of esophageal adenocarcinoma by 29% (odds ratio [OR], 0.71; 95% confidence interval [CI], 0.43 1.17), but not of squamous-cell carcinoma (OR, 1.27; 95% CI, 0.71-2.28) compared with nonusers. A high daily dose of ACE inhibitors decreased the risk of both adenocarcinoma and squamous-cell cancer of the esophagus and rendered a 45% decrease of total esophageal cancer (OR, 0.55; 95% CI, 0.33-0.93). Our data showed no clear association between the use of ACE inhibitors and risk of gastric cancer (OR, 1.07; 95% CI, 0.84-1.36). CONCLUSIONS: The use of ACE inhibitors may decrease the risk of developing esophageal cancer, particularly among users with a high daily dose. No association was found between gastric cancer and ACE inhibitors. PMID- 17916546 TI - Does early colectomy increase desmoid risk in familial adenomatous polyposis? AB - BACKGROUND & AIMS: Desmoid tumors are non-metastasizing fibromatoses that occur in 10%-20% of subjects with familial adenomatous polyposis (FAP). Intra-abdominal desmoid tumors are a major cause of mortality in FAP. FAP-associated desmoid tumors are linked to trauma, particularly abdominal surgery, family history of desmoids, hormonal factors, and the location of the APC mutation. We hypothesized that prophylactic colectomy at an early age might increase the risk of developing desmoids. The aim of this study was to determine whether colectomy earlier in life is a risk factor for the development of desmoid tumors. METHODS: An analysis was made of the association between development of desmoid and age at colectomy, family history of desmoids, gender, and APC mutation in FAP patients in the Registry (1980-2005) at Mount Sinai Hospital, Toronto, Ontario, Canada. RESULTS: FAP patients (n = 930) from 365 kindreds were identified. Desmoid prevalence was 14% (n = 121). Female patients were more likely to develop desmoids than male patients (17% vs 11%, P = .03). Female patients who had an early colectomy were more than 2 times more likely to develop a desmoid, compared with women who had a colectomy at >18 years (P = .01). Early colectomy did not increase risk of developing a desmoid in male patients (P = .42). Female patients who had an early colectomy (/=F2) and cirrhosis (F4) in the training set. Receiver operating characteristic curves were constructed for these factors to evaluate the diagnostic accuracy of significant hepatic fibrosis and cirrhosis in the training set, and in the validation set to evaluate the reproducibility. RESULTS: Multivariate logistic regression revealed that the splenic arterial pulsatility index (SAPI) and the mean portal vein velocity were predictive of significant fibrosis (>/=F2) and cirrhosis (F4). Receiver operating characteristic analysis showed the areas under the curves of regression models and SAPI were comparable in predicting significant fibrosis (0.88 vs 0.87, P = .22) and cirrhosis (0.92 vs 0.90, P = .12) in the training set. Areas under the curves of SAPI were 0.89 and 0.92 in predicting significant hepatic fibrosis and cirrhosis in the validation set. By choosing optimized cut-off levels, 54% and 76% of the patients with significant hepatic fibrosis and cirrhosis could be predicted correctly. CONCLUSIONS: SAPI is accurate and reproducible for assessing the severity of hepatic fibrosis in patients with CHC. Applying this simple Doppler index can decrease the need for staging liver biopsy. PMID- 17916549 TI - Ultrasound-based transient elastography for the detection of hepatic fibrosis: systematic review and meta-analysis. AB - BACKGROUND & AIMS: Ultrasound-based transient elastography is a promising noninvasive alternative to liver biopsy for detecting hepatic fibrosis. However, its overall test performance in various settings remains unknown. The aims of this study were to perform a systematic review and meta-analysis of diagnostic accuracy studies comparing ultrasound-based transient elastography with liver biopsy for hepatic fibrosis. METHODS: Electronic and manual bibliographic searches to identify potential studies were performed. Selection of studies was based on reported accuracy of ultrasound-based transient elastography compared with liver biopsy. Data extraction was performed independently by 2 reviewers. Meta-analysis combined the sensitivities, specificities, and likelihood ratios of individual studies. Extent and reasons for heterogeneity were assessed. RESULTS: Nine studies in full publication were identified. For patients with stage IV fibrosis (cirrhosis), the pooled estimates for sensitivity were 87% (95% confidence interval [CI], 84%-90%), specificity 91% (95% CI, 89%-92%), positive likelihood ratio 11.7 (95% CI, 7.9-17.1), and negative likelihood ratio 0.14 (95% CI, 0.10-0.20). Among 7 investigations reporting patients with stages II-IV fibrosis, the pooled estimates for sensitivity were 70% (95% CI, 67%-73%), specificity 84% (95% CI, 80%-88%), positive likelihood ratio 4.2 (95% CI, 2.4 7.2), and negative likelihood ratio 0.31 (95% CI, 0.23-0.43). Diagnostic threshold (or cut-off value) bias was identified as an important cause of heterogeneity for pooled results in both patient groups. CONCLUSIONS: Ultrasound based transient elastography appears to be a clinically useful test for detecting cirrhosis. PMID- 17916548 TI - Assessment of hepatic fibrosis with magnetic resonance elastography. AB - BACKGROUND & AIMS: Accurate detection of hepatic fibrosis is crucial for assessing prognosis and candidacy for treatment in patients with chronic liver disease. Magnetic resonance (MR) elastography, a technique for quantitatively assessing the mechanical properties of soft tissues, has been shown previously to have potential for noninvasively detecting liver fibrosis. The goal of this work was to obtain preliminary estimates of the sensitivity and specificity of the technique in diagnosing liver fibrosis, and to assess its potential for identifying patients who potentially can avoid a biopsy procedure. METHODS: MR elastography was performed in 35 normal volunteers and 50 patients with chronic liver disease. MR imaging measurements of hepatic fat to water ratios were obtained to assess the potential for fat infiltration to affect stiffness-based detection of fibrosis. RESULTS: Liver stiffness increased systematically with fibrosis stage. Receiver operating curve analysis showed that, with a shear stiffness cut-off value of 2.93 kilopascals, the predicted sensitivity and specificity for detecting all grades of liver fibrosis is 98% and 99%, respectively. Receiver operating curve analysis also provided evidence that MR elastography can discriminate between patients with moderate and severe fibrosis (grades 2-4) and those with mild fibrosis (sensitivity, 86%; specificity, 85%). Hepatic stiffness does not appear to be influenced by the degree of steatosis. CONCLUSIONS: MR elastography is a safe, noninvasive technique with excellent diagnostic accuracy for assessing hepatic fibrosis. Based on the high negative predictive value of MR elastography, an initial clinical application may be to triage patients who are under consideration for biopsy examination to assess possible hepatic fibrosis. PMID- 17916550 TI - Quantifying social distancing arising from pandemic influenza. AB - Local epidemic curves during the 1918-1919 influenza pandemic were often characterized by multiple epidemic waves. Identifying the underlying cause(s) of such waves may help manage future pandemics. We investigate the hypothesis that these waves were caused by people avoiding potentially infectious contacts-a behaviour termed 'social distancing'. We estimate the effective disease reproduction number and from it infer the maximum degree of social distancing that occurred during the course of the multiple-wave epidemic in Sydney, Australia. We estimate that, on average across the city, people reduced their infectious contact rate by as much as 38%, and that this was sufficient to explain the multiple waves of this epidemic. The basic reproduction number, R0, was estimated to be in the range of 1.6-2.0 with a preferred estimate of 1.8, in line with other recent estimates for the 1918-1919 influenza pandemic. The data are also consistent with a high proportion (more than 90%) of the population being initially susceptible to clinical infection, and the proportion of infections that were asymptomatic (if this occurs) being no higher than approximately 9%. The observed clinical attack rate of 36.6% was substantially lower than the 59% expected based on the estimated value of R0, implying that approximately 22% of the population were spared from clinical infection. This reduction in the clinical attack rate translates to an estimated 260 per 100000 lives having been saved, and suggests that social distancing interventions could play a major role in mitigating the public health impact of future influenza pandemics. PMID- 17916551 TI - An analysis of the link between behavioural, biological and social risk factors and subsequent hospital admission in Scotland. AB - OBJECTIVE: To determine the association between risk factors and hospital admission. METHODS: The 1998 Scottish Health Survey was linked to the Scottish hospital admission database. FINDINGS: Smoking was the most important behavioural risk factor (hazard ratio: 1.90, 95% CI: 1.59-2.27). Other behavioural risk factors yielded small but largely anticipated results. Hazard ratios for biological risks increased predictably but with some exceptions (blood pressure and total cholesterol). The top quintile for C-reactive protein showed almost double the risk of admission compared with the bottom quintile (hazard ratio: 1.93, 95% CI: 1.52-2.46). Elevated body mass index (BMI) increased the risk of serious admission (hazard ratio: 1.23, 95% CI: 1.03-1.47) and raised gamma-GT increased this risk by 20% (hazard ratio: 1.20, 95% CI: 1.04-1.38). Forced expiratory volume was the 'biological' factor with the largest risk (hazard ratio for lowest category: 1.82, 95% CI: 1.49-2.22). All the measures of social position showed variable effects on the risk of hospital admission. Large effects on risk were associated with self assessed health, longstanding illness and previous admission. CONCLUSION: The linkage of national surveys with a prospective hospitalization database will develop into an increasingly powerful tool. PMID- 17916552 TI - Piggy-back pacing: implantation of pacemaker and defibrillator on top of each other. AB - Following the publication of several landmark trials, there has been a significant increase in the cardiac device implantation. Within this population there are a number of patients who have pre-existing cardiac devices that have been placed for a number of different conditions. While the usual approach is to remove the existing unit and replace it with a new device with the removal or capping of existing lead systems, this practice often sacrifices an existing unit that still possesses good battery longevity. We explored the possibility of separating the pacing and defibrillating functions by implanting a new device on the top of the old device in a 'piggy-back fashion'. We report a series of four cases (with various indications) with differing combinations of devices. The procedure was performed safely in every one of them, and no device-device interaction was noted. Combining the new with existing units in a 'piggy-back' manner may be a safe and cost-effective technique in the selected cases. PMID- 17916553 TI - A subpopulation of macrophages infiltrates hypertrophic adipose tissue and is activated by free fatty acids via Toll-like receptors 2 and 4 and JNK-dependent pathways. AB - Obesity and type 2 diabetes are characterized by decreased insulin sensitivity, elevated concentrations of free fatty acids (FFAs), and increased macrophage infiltration in adipose tissue (AT). Here, we show that FFAs can cause activation of RAW264.7 cells primarily via the JNK signaling cascade and that TLR2 and TLR4 are upstream of JNK and help transduce FFA proinflammatory signals. We also demonstrate that F4/80(+)CD11b(+)CD11c(+) bone marrow-derived dendritic cells (BMDCs) have heightened proinflammatory activity compared with F4/80(+)CD11b(+)CD11c(-) bone marrow-derived macrophages and that the proinflammatory activity and JNK phosphorylation of BMDCs, but not bone marrow derived macrophages, was further increased by FFA treatment. F4/80(+)CD11b(+)CD11c(+) cells were found in AT, and the proportion and number of these cells in AT is increased in ob/ob mice and by feeding wild type mice a high fat diet for 1 and 12 weeks. AT F4/80(+)CD11b(+)CD11c(+) cells express increased inflammatory markers compared with F4/80(+)CD11b(+)CD11c(-) cells, and FFA treatment increased inflammatory responses in these cells. In addition, we found that CD11c expression is increased in skeletal muscle of high fat diet-fed mice and that conditioned medium from FFA-treated wild type BMDCs, but not TLR2/4 DKO BMDCs, can induce insulin resistance in L6 myotubes. Together our results show that FFAs can activate CD11c(+) myeloid proinflammatory cells via TLR2/4 and JNK signaling pathways, thereby promoting inflammation and subsequent cellular insulin resistance. PMID- 17916554 TI - Structural basis for the recognition and cross-linking of amyloid fibrils by human apolipoprotein E. AB - Apolipoprotein (apo) E is a well characterized lipid-binding protein in plasma that also exists as a common nonfibrillar component of both cerebral and systemic amyloid deposits. A genetic link between a common isoform of apoE, apoE4, and the incidence of late onset Alzheimer disease has drawn considerable attention to the potential roles of apoE in amyloid-related disease. We examined the interactions of apoE with amyloid fibrils composed of apoC-II and the amyloid-beta (Abeta) peptide. Aggregates of apoE with Abeta and apoC-II are found in Alzheimer and atherosclerotic plaques, respectively. Sedimentation velocity and fibril size distribution analysis showed that apoE3 and E4 isoforms bind and noncovalently cross-link apoC-II fibrils in a similar manner. This ability to cross-link apoC II fibrils was abolished by the dissociation of the apoE tetramer to monomers or by thrombin cleavage to yield separate N- and C-terminal domains. Preparative ultracentrifuge binding studies indicated that apoE and the isolated N- and C terminal domains of apoE bind with submicromolar affinities to both apoC-II and Abeta fibrils. Fluorescence quenching and resonance energy transfer experiments confirmed that both domains of apoE interact with apoC-II fibrils and demonstrated that the binding of the isolated N-terminal domain of apoE to apoC II or Abeta fibrils is accompanied by a significant conformational change with helix three of the domain moving relative to helix one. We propose a model involving the interaction of apoE with patterns of aligned residues that could explain the general ability of apoE to bind to a diverse range of amyloid fibrils. PMID- 17916555 TI - Butyl isocyanide as a probe of the activation mechanism of soluble guanylate cyclase. Investigating the role of non-heme nitric oxide. AB - Nitric oxide (NO) is a physiologically relevant activator of the hemoprotein soluble guanylate cyclase (sGC). In the presence of NO, sGC is activated several hundredfold above the basal level by a mechanism that remains to be elucidated. The heme ligand n-butyl isocyanide (BIC) was used to probe the mechanism of NO activation of sGC. Electronic absorption spectroscopy was used to show that BIC binds to the sGC heme, forming a 6-coordinate complex with an absorbance maximum at 429 nm. BIC activates sGC 2-5-fold, and synergizes with the allosteric activator YC-1, to activate the enzyme 15-25-fold. YC-1 activates the sGC-BIC complex, and leads to an increase in both the V(max) and K(m). BIC was also used to probe the mechanism of NO activation. The activity of the sGC-BIC complex increases 15-fold in the presence of NO, without displacing BIC at the heme, which is consistent with previous reports that proposed the involvement of a non heme NO binding site in the activation process. PMID- 17916556 TI - Crystal structures of yeast beta-alanine synthase complexes reveal the mode of substrate binding and large scale domain closure movements. AB - Beta-alanine synthase is the final enzyme of the reductive pyrimidine catabolic pathway, which is responsible for the breakdown of uracil and thymine in higher organisms. The fold of the homodimeric enzyme from the yeast Saccharomyces kluyveri identifies it as a member of the AcyI/M20 family of metallopeptidases. Its subunit consists of a catalytic domain harboring a di-zinc center and a smaller dimerization domain. The present site-directed mutagenesis studies identify Glu(159) and Arg(322) as crucial for catalysis and His(262) and His(397) as functionally important but not essential. We determined the crystal structures of wild-type beta-alanine synthase in complex with the reaction product beta alanine, and of the mutant E159A with the substrate N-carbamyl-beta-alanine, revealing the closed state of a dimeric AcyI/M20 metallopeptidase-like enzyme. Subunit closure is achieved by a approximately 30 degrees rigid body domain rotation, which completes the active site by integration of substrate binding residues that belong to the dimerization domain of the same or the partner subunit. Substrate binding is achieved via a salt bridge, a number of hydrogen bonds, and coordination to one of the zinc ions of the di-metal center. PMID- 17916557 TI - A smooth muscle Cav1.2 calcium channel splice variant underlies hyperpolarized window current and enhanced state-dependent inhibition by nifedipine. AB - Native smooth muscle L-type Ca(v)1.2 calcium channels have been shown to support a fraction of Ca(2+) currents with a window current that is close to resting potential. The smooth muscle L-type Ca(2+) channels are also more susceptible to inhibition by dihydropyridines (DHPs) than the cardiac channels. It was hypothesized that smooth muscle Ca(v)1.2 channels exhibiting hyperpolarized shift in steady-state inactivation would contribute to larger inhibition by DHP, in addition to structural differences of the channels generated by alternative splicing that modulate DHP sensitivities. In addition, it has also been shown that alternative splicing modulates DHP sensitivities by generating structural differences in the Ca(v)1.2 channels. Here, we report a smooth muscle L-type Ca(v)1.2 calcium channel splice variant, Ca(v)1.2SM (1/8/9(*)/32/Delta33), that when expressed in HEK 293 cells display hyperpolarized shifts for steady-state inactivation and activation potentials when compared with the established Ca(v)1.2b clone (1/8/9(*)/32/33). This variant activates from more negative potentials and generates a window current closer to resting membrane potential. We also identified the predominant cardiac isoform Ca(v)1.2CM clone (1a/8a/Delta9(*)/32/33) that is different from the established Ca(v)1.2a (1a/8a/Delta9(*)/31/33). Importantly, Ca(v)1.2SM channels were shown to be more sensitive to nifedipine blockade than Ca(v)1.2b and cardiac Ca(v)1.2CM channels when currents were recorded in either 5 mM Ba(2+) or 1.8 mM Ca(2+) external solutions. This is the first time that a smooth muscle Ca(v)1.2 splice variant has been identified functionally to possess biophysical property that can be linked to enhanced state-dependent block by DHP. PMID- 17916558 TI - Deletion of the aryl hydrocarbon receptor-associated protein 9 leads to cardiac malformation and embryonic lethality. AB - The aryl hydrocarbon receptor-associated protein 9, ARA9 (also known as XAP2 or AIP1), is a chaperone that is found in complexes with certain xenobiotic receptors, such as the aryl hydrocarbon receptor (AHR) and the peroxisome proliferator-activated receptor alpha (PPARalpha). In an effort to better understand the physiological role of ARA9 outside of its role in xenobiotic signal transduction, we generated a null allele at the Ara9 locus in mice. Mice with a homozygous deletion of this gene die at various time points throughout embryonic development. Embryonic lethality is accompanied by decreased blood flow to head and limbs, as well as a range of heart deformations, including double outlet right ventricle, ventricular-septal defects, and pericardial edema. The early cardiovascular defects observed in Ara9-null mice suggest an essential role for the ARA9 protein in cardiac development. The observation that the developmental aberrations in Ara9-null mice are distinct from those observed for disrupted alleles at Ahr or Pparalpha indicates that the role of ARA9 in cardiac development is independent of its interactions with its known xenobiotic receptor partners. PMID- 17916559 TI - Sts1 can overcome the loss of Rad23 and Rpn10 and represents a novel regulator of the ubiquitin/proteasome pathway. AB - A rad23Delta rpn10Delta double mutant accumulates multi-Ub proteins, is deficient in proteolysis, and displays sensitivity to drugs that generate damaged proteins. Overexpression of Sts1 restored normal growth in rad23Delta rpn10Delta but did not overcome the DNA repair defect of rad23Delta. To understand the nature of Sts1 suppression, we characterized sts1-2, a temperature-sensitive mutant. We determined that sts1-2 was sensitive to translation inhibitors, accumulated high levels of multi-Ub proteins, and caused stabilization of proteolytic substrates. Additionally, ubiquitinated proteins that were detected in proteasomes were inefficiently cleared in sts1-2. Despite these proteolytic defects, overall proteasome activity was increased in sts1-2. We propose that Sts1 is a new regulatory factor in the ubiquitin/proteasome pathway that controls the turnover of proteasome substrates. PMID- 17916560 TI - Binding of Rac1, Rnd1, and RhoD to a novel Rho GTPase interaction motif destabilizes dimerization of the plexin-B1 effector domain. AB - Plexins are the first known transmembrane receptors that interact directly with small GTPases. On binding to certain Rho family GTPases, the receptor regulates the remodeling of the actin cytoskeleton and alters cell movement in response to semaphorin guidance cues. In a joint solution NMR spectroscopy and x-ray crystallographic study, we characterize a 120-residue cytoplasmic independent folding domain of plexin-B1 that directly binds three Rho family GTPases, Rac1, Rnd1, and RhoD. The NMR data show that, surprisingly, the Cdc42/Rac interactive binding-like motif of plexin-B1 is not involved in this interaction. Instead, all three GTPases interact with the same region, beta-strands 3 and 4 and a short alpha-helical segment of the plexin domain. The 2.0 A resolution x-ray structure shows that these segments are brought together by the tertiary structure of the ubiquitin-like fold. In the crystal, the protein is dimerized with C2 symmetry through a four-stranded antiparallel beta-sheet that is formed outside the fold by a long loop between the monomers. This region is adjacent to the GTPase binding motifs identified by NMR. Destabilization of the dimer in solution by binding of any one of the three GTPases suggests a model for receptor regulation that involves bidirectional signaling. The model implies a multifunctional role for the GTPase-plexin interaction that includes conformational change and a localization of active receptors in the signaling mechanism. PMID- 17916561 TI - Plasma membrane rafts complete cholesterol synthesis by participating in retrograde movement of precursor sterols. AB - Mammalian cells synthesize significant amounts of precursor sterols, in addition to cholesterol, at the endoplasmic reticulum (ER). The newly synthesized sterols rapidly move to the plasma membrane (PM). The mechanism by which precursor sterols move back to the ER for their enzymatic processing to cholesterol is essentially unknown. Here we performed pulse-chase experiments and showed that the C29/C30 sterols rapidly move from the PM to the ER and are converted to cholesterol. The retrograde precursor sterol transport is largely independent of the Niemann-Pick type C proteins, which play important roles in late endosomal cholesterol transport. In contrast, disrupting lipid rafts significantly retards the conversion of C29/C30 and C28 sterols to cholesterol, causing the accumulation of precursor sterols at the PM. Our results reveal a previously undisclosed function of the PM lipid rafts: they bring cholesterol biosynthesis to completion by participating in the retrograde movement of precursor sterols back to the ER. PMID- 17916562 TI - Dynamics of the hypoxia-inducible factor-1-vascular endothelial growth factor promoter complex. AB - Some transactivator-promoter complexes are highly dynamic due to active disruption of the complex by proteolytic or nonproteolytic mechanisms, and this appears to be an important mechanism by which their activity is governed tightly and eventually terminated. However, the generality of these mechanisms is unclear. In this report, we address the dynamics of hypoxia-inducible factor-1 (HIF-1) binding to the vascular endothelial growth factor promoter. HIF-1 is a heterodimeric transcription factor whose activity is triggered by an increase in HIF-1alpha levels in hypoxic cells. A "competition ChIP" assay is employed to demonstrate that HIF-1alpha forms a kinetically stable complex with the native vascular endothelial growth factor promoter that has a half-life in excess of 1 h. Thus, HIF-1 activity does not require rapid proteolytic turnover of the promoter-bound transactivator, nor is the activator-promoter complex constantly disassembled by chaperones. However, we do find that after cessation of the inducing signal, HIF-1 activity is slowly returned to basal levels by proteasome mediated proteolysis of the promoter-bound HIF-1alpha protein. PMID- 17916563 TI - NM23-H1 tumor suppressor and its interacting partner STRAP activate p53 function. AB - p53 plays a critical role in a variety of growth inhibitory responses, including cell cycle arrest, differentiation, and apoptosis, and contributes to tumor suppression. Here we show that NM23-H1 and its binding partner STRAP (serine threonine kinase receptor-associated protein) interact with p53 and potentiate p53 activity. Both NM23-H1 and STRAP directly interact with the central DNA binding domain within residues 113-290. The use of NM23-H1 and STRAP mutants revealed that Cys(145) of NM23-H1 and Cys(152) (or Cys(270)) of STRAP were responsible for p53 binding. Furthermore, Cys(176) and Cys(135) of p53 were required to bind NM23-H1 and STRAP, respectively. Ectopic expression of wild-type NM23-H1 and STRAP, but not NM23-H1(C145S) and STRAP(C152S/C270S), positively regulated p53-mediated transcription in a dose-dependent manner. Knockdown of endogenous NM23-H1 or STRAP produced an opposite trend and inhibited the p53 mediated transcription. Similarly, NM23-H1 and STRAP stimulated p53-induced apoptosis and growth inhibition, whereas the NM23-H1(C145S) and STRAP(C152S/C270S) mutants had no effect. We also demonstrated that p53 activation by NM23-H1 and STRAP was mediated by removing Mdm2, a negative regulator of p53, from the p53-Mdm2 complex. These results suggest that NM23-H1 and its interacting partner STRAP physically interact with p53 and positively regulate its functions, including p53-induced apoptosis and cell cycle arrest. PMID- 17916568 TI - Morphology and meiotic segregation in spermatozoa from men of proven fertility. AB - Estimates of semen parameters are important for defining normal ranges, which are currently established by 1999 World Health Organization guidelines. However, it is well known that semen evaluation is questionable because it is necessary for only 1 sperm to be able to reach and fertilize the oocyte. Spermiogram parameters and sperm morphology, evaluated by transmission electron microscopy (TEM), were performed on semen samples from 25 men of proven fertility. Despite a generally normal sperm concentration, progressive motility was reduced in 9 cases. Sperm characteristics were evaluated with an established technique, and the mean of the percentages of sperm pathologies were confirmed by comparing to previous reports. A comparison of apoptosis and necrosis in the samples, as detected by TEM and an annexin V/propidium iodide assay, was also performed. Fluorescence in situ hybridization was carried out on the same samples using probes for chromosomes 18, X, and Y. The mean value of the frequency of total aneuploidy in the analyzed group was 0.627% (25th percentile = 0.563%; median = 0.625%; 75th percentile = 0.690%). This study of the incidence of disomy and diploidy in spermatozoa from fertile, apparently normal individuals is important for making comparisons with infertile cohorts to determine the real increase of aneuploidy in those cohorts. PMID- 17916567 TI - Downregulation of thymosin beta4 expression by androgen in prostate cancer LNCaP cells. AB - Androgen ablation therapy is an effective treatment for advanced prostate cancer, but the tumor often progresses toward a more aggressive phenotype. We determined the changes in genes associated with the malignant progression and found increased thymosin beta4, involved in tumor metastasis, in androgen-sensitive LNCaP cells grown in the medium with androgen-deficient, charcoal-stripped fetal calf serum. The mRNA expression of thymosin beta4 was determined by real-time polymerase chain reaction analysis. The transcriptional activity of thymosin beta4 was measured by luciferase assay using reporter plasmid containing 5' flanking region of thymosin beta4. Thymosin beta4 mRNA expression was increased in LNCaP cells in the androgen-deficient condition and decreased by dihydrotestosterone treatment. Androgen receptor antagonist bicalutamide inhibited thymosin beta4 expression in a dose-dependent manner. In androgen receptor-negative PC-3 cells, no significant effects on thymosin beta4 gene expression were observed. The regulation of thymosin beta4 mRNA expression by androgen is due to the transcriptional activation. Deletion analysis revealed that the region between -83 bp and -46 bp of the thymosin beta4 gene is responsible for the regulation of the transcriptional activity by androgen. Thymosin beta4 expression is negatively controlled at the transcriptional level by androgen. PMID- 17916569 TI - A dose-response study of testosterone on sexual dysfunction and features of the metabolic syndrome using testosterone gel and parenteral testosterone undecanoate. AB - The objective of this study was to observe the dose-response effects of testosterone (T) treatment on symptoms of sexual dysfunction and the metabolic syndrome. Two cohorts of elderly men with late-onset hypogonadism were followed over 9 months. Group 1, consisting of 28 men (mean age, 61 years; mean T level, 2.07 +/- 0.50 ng/mL), received long-acting T undecanoate (TU; 1000 mg); group 2, composed of 27 men (mean age, 60 years; mean T level, 2.24 +/- 0.41 ng/mL), received T gel (50 mg/day) for 9 months. In patients treated with T gel, plasma T levels rose from 2.24 +/- 0.41 to 2.95 +/- 0.52 (statistically significant) at 3 months, 3.49 +/- 0.89 (statistically significant) at 6 months, and 3.80 +/- 0.73 ng/mL at 9 months (T level at 6 months was compared with T level at 3 months). With TU, plasma T levels rose from 2.08 +/- 0.56 to 4.81 +/- 0.83 (statistically significant) at 3 months, 5.29 +/- 0.91 at 6 months, and 5.40 +/- 0.77 ng/mL at 9 months. With TU, the plasma T levels were statistically significantly higher than with T gel With TU, there was a greater improvement in sexual symptoms and in symptoms of the metabolic syndrome. With both treatments, changes in waist circumference correlated with changes in total, low-density, and high-density lipoprotein cholesterol. Parameters of safety were not different between the 2 treatments. T administration had a beneficial effect on sexual dysfunction and symptoms of the metabolic syndrome in elderly men. The higher plasma levels of T generated with TU than with T gel were clearly more effective, indicating that there is a T dose-effect relationship. PMID- 17916570 TI - PPT-DB: the protein property prediction and testing database. AB - The protein property prediction and testing database (PPT-DB) is a database housing nearly 30 carefully curated databases, each of which contains commonly predicted protein property information. These properties include both structural (i.e. secondary structure, contact order, disulfide pairing) and dynamic (i.e. order parameters, B-factors, folding rates) features that have been measured, derived or tabulated from a variety of sources. PPT-DB is designed to serve two purposes. First it is intended to serve as a centralized, up-to-date, freely downloadable and easily queried repository of predictable or 'derived' protein property data. In this role, PPT-DB can serve as a one-stop, fully standardized repository for developers to obtain the required training, testing and validation data needed for almost any kind of protein property prediction program they may wish to create. The second role that PPT-DB can play is as a tool for homology based protein property prediction. Users may query PPT-DB with a sequence of interest and have a specific property predicted using a sequence similarity search against PPT-DB's extensive collection of proteins with known properties. PPT-DB exploits the well-known fact that protein structure and dynamic properties are highly conserved between homologous proteins. Predictions derived from PPT DB's similarity searches are typically 85-95% correct (for categorical predictions, such as secondary structure) or exhibit correlations of >0.80 (for numeric predictions, such as accessible surface area). This performance is 10-20% better than what is typically obtained from standard 'ab initio' predictions. PPT DB, its prediction utilities and all of its contents are available at http://www.pptdb.ca. PMID- 17916571 TI - 4DXpress: a database for cross-species expression pattern comparisons. AB - In the major animal model species like mouse, fish or fly, detailed spatial information on gene expression over time can be acquired through whole mount in situ hybridization experiments. In these species, expression patterns of many genes have been studied and data has been integrated into dedicated model organism databases like ZFIN for zebrafish, MEPD for medaka, BDGP for Drosophila or GXD for mouse. However, a central repository that allows users to query and compare gene expression patterns across different species has not yet been established. Therefore, we have integrated expression patterns for zebrafish, Drosophila, medaka and mouse into a central public repository called 4DXpress (expression database in four dimensions). Users can query anatomy ontology-based expression annotations across species and quickly jump from one gene to the orthologues in other species. Genes are linked to public microarray data in ArrayExpress. We have mapped developmental stages between the species to be able to compare developmental time phases. We store the largest collection of gene expression patterns available to date in an individual resource, reflecting 16 505 annotated genes. 4DXpress will be an invaluable tool for developmental as well as for computational biologists interested in gene regulation and evolution. 4DXpress is available at http://ani.embl.de/4DXpress. PMID- 17916572 TI - Greglist: a database listing potential G-quadruplex regulated genes. AB - The double helix is a conformation that genomic DNA usually assumes; under certain conditions, however, guanine-rich DNA sequences can form a four-stranded structure, G-quadruplex, which is found to play a role in regulating gene expression. Indeed, it has been demonstrated that the G-quadruplex formed in the c-MYC promoter suppresses its transcriptional activity. Recent studies suggest that G-quadruplex motifs (GQMs) are enriched in human gene promoters. To facilitate the research of G-quadruplex, we have constructed Greglist, a database listing potentially G-quadruplex regulated genes. Greglist harbors genes that contain promoter GQMs from genomes of various species, including humans, mice, rats and chickens. Many important genes are found to contain previously unreported promoter GQMs, such as ATM, BAD, AKT1, LEPR, UCP1, APOE, DKK1, WT1, WEE1, WNT1 and CLOCK. Furthermore, we find that not only protein coding genes, 126 human microRNAs also contain promoter GQMs. Greglist therefore provides candidates for further studying G-quadruplex functions and is freely available at http://tubic.tju.edu.cn/greglist. PMID- 17916573 TI - DB-PABP: a database of polyanion-binding proteins. AB - The interactions between polyanions (PAs) and polyanion-binding proteins (PABPs) have been found to play significant roles in many essential biological processes including intracellular organization, transport and protein folding. Furthermore, many neurodegenerative disease-related proteins are PABPs. Thus, a better understanding of PA/PABP interactions may not only enhance our understandings of biological systems but also provide new clues to these deadly diseases. The literature in this field is widely scattered, suggesting the need for a comprehensive and searchable database of PABPs. The DB-PABP is a comprehensive, manually curated and searchable database of experimentally characterized PABPs. It is freely available and can be accessed online at http://pabp.bcf.ku.edu/DB_PABP/. The DB-PABP was implemented as a MySQL relational database. An interactive web interface was created using Java Server Pages (JSP). The search page of the database is organized into a main search form and a section for utilities. The main search form enables custom searches via four menus: protein names, polyanion names, the source species of the proteins and the methods used to discover the interactions. Available utilities include a commonality matrix, a function of listing PABPs by the number of interacting polyanions and a string search for author surnames. The DB-PABP is maintained at the University of Kansas. We encourage users to provide feedback and submit new data and references. PMID- 17916574 TI - Shanghai RAPESEED Database: a resource for functional genomics studies of seed development and fatty acid metabolism of Brassica. AB - The Shanghai RAPESEED Database (RAPESEED, http://rapeseed.plantsignal.cn/) was created to provide the solid platform for functional genomics studies of oilseed crops with the emphasis on seed development and fatty acid metabolism. The RAPESEED includes the resource of 8462 unique ESTs, of which 3526 clones are with full length cDNA; the expression profiles of 8095 genes and the Serial Analysis of Gene Expression (SAGE, 23,895 unique tags) and tag-to-gene data during seed development. In addition, a total of approximately 14,700 M3 mutant populations were generated by ethylmethanesulfonate (EMS) mutagenesis and related seed quality information was determined using the Foss NIR System. Further, the TILLING (Targeting Induced Local Lesions IN Genomes) platform was established based on the generated EMS mutant population. The relevant information was collected in RAPESEED database, which can be searched through keywords, nucleotide or protein sequences, or seed quality parameters, and downloaded. PMID- 17916575 TI - Discovery of a mRNA mitochondrial localization element in Saccharomyces cerevisiae by nonhomologous random recombination and in vivo selection. AB - In budding yeast, over 100 nuclear-encoded mRNAs are localized to the mitochondria. The determinants of mRNA localization to the mitochondria are not well understood, and protein factors involved in this process have not yet been identified. To reveal the sequence determinants for mitochondrial localization in a comprehensive and unbiased manner, we generated highly diversified libraries of 3' UTR regions from a known mitochondrially localized mRNA by nonhomologous random recombination (NRR) and subjected the resulting sequences to an in vivo selection that links cell survival to mitochondrial mRNA localization. When applied to the yeast ATP2 mRNA, this approach rapidly identified a 50-nt consensus motif, designated Min2, as well as two Min2-homologous regions naturally present downstream of the ATP2 stop codon, which are sufficient when appended to the 3' end of various reporter mRNAs to induce mitochondrial localization. Site-directed mutagenesis of Min2 revealed primary and secondary structure elements that contribute to localization activity. In addition, the Min2 motif may facilitate the identification of proteins involved in this mode of establishing cellular asymmetry. PMID- 17916576 TI - C to U editing at position 32 of the anticodon loop precedes tRNA 5' leader removal in trypanosomatids. AB - In all organisms, precursor tRNAs are processed into mature functional units by post-transcriptional changes. These involve 5' and 3' end trimming as well as the addition of a significant number of chemical modifications, including RNA editing. The only known example of non-organellar C to U editing of tRNAs occurs in trypanosomatids. In this system, editing at position 32 of the anticodon loop of tRNA(Thr)(AGU) stimulates, but is not required for, the subsequent formation of inosine at position 34. In the present work, we expand the number of C to U edited tRNAs to include all the threonyl tRNA isoacceptors. Notably, the absence of a naturally encoded adenosine, at position 34, in two of these isoacceptors demonstrates that A to I is not required for C to U editing. We also show that C to U editing is a nuclear event while A to I is cytoplasmic, where C to U editing at position 32 occurs in the precursor tRNA prior to 5' leader removal. Our data supports the view that C to U editing is more widespread than previously thought and is part of a stepwise process in the maturation of tRNAs in these organisms. PMID- 17916577 TI - The small RNA repertoire of Dictyostelium discoideum and its regulation by components of the RNAi pathway. AB - Small RNAs play crucial roles in regulation of gene expression in many eukaryotes. Here, we report the cloning and characterization of 18-26 nt RNAs in the social amoeba Dictyostelium discoideum. This survey uncovered developmentally regulated microRNA candidates whose biogenesis, at least in one case, is dependent on a Dicer homolog, DrnB. Furthermore, we identified a large number of 21 nt RNAs originating from the DIRS-1 retrotransposon, clusters of which have been suggested to constitute centromeres. Small RNAs from another retrotransposon, Skipper, were significantly up-regulated in strains depleted of the second Dicer-like protein, DrnA, and a putative RNA-dependent RNA polymerase, RrpC. In contrast, the expression of DIRS-1 small RNAs was not altered in any of the analyzed strains. This suggests the presence of multiple RNAi pathways in D. discoideum. In addition, we isolated several small RNAs with antisense complementarity to mRNAs. Three of these mRNAs are developmentally regulated. Interestingly, all three corresponding genes express longer antisense RNAs from which the small RNAs may originate. In at least one case, the longer antisense RNA is complementary to the spliced but not the unspliced pre-mRNA, indicating synthesis by an RNA-dependent RNA polymerase. PMID- 17916578 TI - Escherichia coli HU protein has a role in the repair of abasic sites in DNA. AB - HU is one of the most abundant DNA binding proteins in Escherichia coli. We find that it binds strongly to DNA containing an abasic (AP) site or tetrahydrofuran (THF) (apparent K(d) approximately 50 nM). It also possesses an AP lyase activity that cleaves at a deoxyribose but not at a THF residue. The binding and cleavage of an AP site was observed only with the HUalphabeta heterodimer. Site-specific mutations at K3 and R61 residues led to a change in substrate binding and cleavage. Both K3A(alpha)K3A(beta) and R61A(alpha)R61A(beta) mutant HU showed significant reduction in binding to DNA containing AP site; however, only R61A(alpha)R61A(beta) mutant protein exhibited significant loss in AP lyase activity. Both K3A(alpha)K3A(beta) and R61K(alpha)R61K(beta) showed slight reduction in AP lyase activities. The function of HU protein as an AP lyase was confirmed by the ability of hupA or hupB mutations to further reduce the viability of an E. coli dut(Ts) xth mutant, which generates lethal AP sites at 37 degrees C; the hupA and hupB derivatives, respectively, had a 6-fold and a 150 fold lower survival at 37 degrees C than did the parental strain. These data suggest, therefore, that HU protein plays a significant role in the repair of AP sites in E. coli. PMID- 17916579 TI - Site-specific interaction of the murine pre-replicative complex with origin DNA: assembly and disassembly during cell cycle transit and differentiation. AB - Eukaryotic DNA replication initiates at origins of replication by the assembly of the highly conserved pre-replicative complex (pre-RC). However, exact sequences for pre-RC binding still remain unknown. By chromatin immunoprecipitation we identified in vivo a pre-RC-binding site within the origin of bidirectional replication in the murine rDNA locus. At this sequence, ORC1, -2, -4 and -5 are bound in G1 phase and at the G1/S transition. During S phase, ORC1 is released. An ATP-dependent and site-specific assembly of the pre-RC at origin DNA was demonstrated in vitro using partially purified murine pre-RC proteins in electrophoretic mobility shift assays. By deletion experiments the sequence required for pre-RC binding was confined to 119 bp. Nucleotide substitutions revealed that two 9 bp sequence elements, CTCGGGAGA, are essential for the binding of pre-RC proteins to origin DNA within the murine rDNA locus. During myogenic differentiation of C2C12 cells, we demonstrated a reduction of ORC1 and ORC2 by immunoblot analyses. ChIP analyses revealed that ORC1 completely disappears from chromatin of terminally differentiated myotubes, whereas ORC2, -4 and -5 still remain associated. PMID- 17916580 TI - Comparative metagenomics revealed commonly enriched gene sets in human gut microbiomes. AB - Numerous microbes inhabit the human intestine, many of which are uncharacterized or uncultivable. They form a complex microbial community that deeply affects human physiology. To identify the genomic features common to all human gut microbiomes as well as those variable among them, we performed a large-scale comparative metagenomic analysis of fecal samples from 13 healthy individuals of various ages, including unweaned infants. We found that, while the gut microbiota from unweaned infants were simple and showed a high inter-individual variation in taxonomic and gene composition, those from adults and weaned children were more complex but showed a high functional uniformity regardless of age or sex. In searching for the genes over-represented in gut microbiomes, we identified 237 gene families commonly enriched in adult-type and 136 families in infant-type microbiomes, with a small overlap. An analysis of their predicted functions revealed various strategies employed by each type of microbiota to adapt to its intestinal environment, suggesting that these gene sets encode the core functions of adult and infant-type gut microbiota. By analysing the orphan genes, 647 new gene families were identified to be exclusively present in human intestinal microbiomes. In addition, we discovered a conjugative transposon family explosively amplified in human gut microbiomes, which strongly suggests that the intestine is a 'hot spot' for horizontal gene transfer between microbes. PMID- 17916581 TI - Classification of the cardiomyopathies: a position statement from the European Society Of Cardiology Working Group on Myocardial and Pericardial Diseases. AB - In biology, classification systems are used to promote understanding and systematic discussion through the use of logical groups and hierarchies. In clinical medicine, similar principles are used to standardise the nomenclature of disease. For more than three decades, heart muscle diseases have been classified into primary or idiopathic myocardial diseases (cardiomyopathies) and secondary disorders that have similar morphological appearances, but which are caused by an identifiable pathology such as coronary artery disease or myocardial infiltration (specific heart muscle diseases). In this document, The European Society of Cardiology Working Group on Myocardial and Pericardial Diseases presents an update of the existing classification scheme. The aim is to help clinicians look beyond generic diagnostic labels in order to reach more specific diagnoses. PMID- 17916582 TI - In vivo response to high-resolution variation of Tbx1 mRNA dosage. AB - Mouse modeling of haploinsufficiency syndromes and, in general, of syndromes caused by gene dosage imbalance, is often unsatisfactory because loss (or gain) of one copy of the gene of interest is insufficient to recapitulate the disease phenotype. In this study, we use Tbx1 mutants, which model one of the most common haploinsufficiency disorders, the 22q11.2DS/DiGeorge/Velocardiofacial syndrome, to test the feasibility of high resolution dosage manipulation to generate mouse models that more closely resemble the human syndrome. We used nine different genotypes at the Tbx1 locus that are associated with progressively lower mRNA levels in vivo. We show that penetrance and expressivity of different phenotypic features became more severe as the dosage diminished, as expected, but the response was strikingly non-linear, with extreme examples such as neonatal lethality, which changed from 2 to 100% after a dosage reduction of just approximately 16%. Furthermore, heart phenotype variability, extreme in the human syndrome but very limited, or absent, in the standard knockout model, was seen when mRNA level was approximately 20% of normal level, suggesting that there is a threshold level associated with unstable balance, which can be perturbed by chance events. Overall, our data suggest that there are developmental process specific gene dosage thresholds beyond which the phenotype worsens very rapidly with very small mRNA level reductions. PMID- 17916583 TI - Human angiogenin is a neuroprotective factor and amyotrophic lateral sclerosis associated angiogenin variants affect neurite extension/pathfinding and survival of motor neurons. AB - Amyotrophic lateral sclerosis (ALS) is a late onset neurodegenerative disorder affecting upper and lower motor neurons (MNs). The molecular mechanisms underlying ALS are poorly understood. Mutations in SOD1 is one of the known causes of ALS but occur only in a very small number of cases of ALS. Interestingly, mutations in human angiogenin (hANG), a member of the ribonuclease A (RNase A) superfamily known to be involved in neovascularization, have been recently reported in patients with ALS, but the effects of these mutations on MN differentiation and survival has not been investigated. We have used the well characterized pluripotent P19 embryonal carcinoma (EC) cell culture model of neuro-ectodermal differentiation to study the effects of hANG-ALS variants on MN differentiation and survival. Here we report that P19 EC cells induced to differentiate in the presence of hANG and hANG-ALS-associated variants internalize the wild-type and variant proteins. The P19 EC cells differentiate to form neurons but the ability of the neurites to extend and make contacts with neighbouring neurites is compromised when treated with the hANG-ALS variants. In addition, hANG-ALS variants also have a cytotoxic effect on MNs leading to their degeneration. hANG was able to protect neurons from hypoxia-induced cell death, but the variants of hANG implicated in ALS lacked the neuroprotective activity. Our findings show that ANG plays an important role in neurite extension/pathfinding and survival providing a causal link between mutations in hANG and ALS. PMID- 17916584 TI - Xyloglucan endotransglucosylase activity loosens a plant cell wall. AB - BACKGROUND AND AIMS: Plant cells undergo cell expansion when a temporary imbalance between the hydraulic pressure of the vacuole and the extensibility of the cell wall makes the cell volume increase dramatically. The primary cell walls of most seed plants consist of cellulose microfibrils tethered mainly by xyloglucans and embedded in a highly hydrated pectin matrix. During cell expansion the wall stress is decreased by the highly controlled rearrangement of the load-bearing tethers in the wall so that the microfibrils can move relative to each other. Here the effect was studied of a purified recombinant xyloglucan endotransglucosylase/hydrolase (XTH) on the extension of isolated cell walls. METHODS: The epidermis of growing onion (Allium cepa) bulb scales is a one-cell thick model tissue that is structurally and mechanically highly anisotropic. In constant load experiments, the effect of purified recombinant XTH proteins of Selaginella kraussiana on the extension of isolated onion epidermis was recorded. KEY RESULTS: Fluorescent xyloglucan endotransglucosylase (XET) assays demonstrate that exogeneous XTH can act on isolated onion epidermis cell walls. Furthermore, cell wall extension was significantly increased upon addition of XTH to the isolated epidermis, but only transverse to the net orientation of cellulose microfibrils. CONCLUSIONS: The results provide evidence that XTHs can act as cell wall-loosening enzymes. PMID- 17916585 TI - Normal bone mineral content but unfavourable muscle/fat ratio in Klinefelter syndrome. AB - OBJECTIVE: To evaluate body composition and bone mineral content (BMC) in children and adolescents with Klinefelter syndrome (KS). DESIGN: Retrospective cross-sectional study. SETTING: Tertiary endocrine clinic at the University Hospital, Copenhagen. PATIENTS: Eighteen untreated boys with KS and six boys with KS receiving androgen substitution with a median age of 11.0 years (range 4.3 18.6) participated in the study. INTERVENTION: Dual energy x ray absorptiometry and anthropometric measurements were analysed. MAIN OUTCOME MEASURES: Lumbar and whole body BMC, lean body mass (LBM), body fat mass (BFM), body fat percentage (BF%), height and body mass index (BMI) were compared between treated and untreated boys with KS and compared to normal age-matched boys. RESULTS: LBM (untreated -0.3 (-2.4 to +2.1) and treated +1.1 (-1.6 to +2.1)) was normal, while BFM (untreated +0.5 (-1.0 to +2.3), p = 0.02 and treated +1.6 (-0.2 to +2.4), p = 0.01) was significantly increased, all expressed as standard deviation scores. Lumbar bone mineral density (BMD; untreated -0.4 (-3.1 to +0.9) and treated +1.0 (-1.4 to +3.0)) and whole body BMC (untreated +0.1 (-1.8 to +3.3) and treated +1.5 (-1.1 to +2.5)) were normal. CONCLUSION: We found significantly increased BFM and BF% despite normal LBM, suggesting the presence of an unfavourable muscle/fat ratio. Lumbar BMD and whole body BMC were normal. These findings suggest that the unfavourable metabolic profile seen in adult KS may already be present in childhood as evidenced by the increased fat mass, whereas the reported low BMD seems to develop after puberty. PMID- 17916586 TI - Parents' journey through treatment for their child's obesity: a qualitative study. AB - BACKGROUND: Treatment for childhood obesity is characterised by patient non attendance and drop-out, and widespread failure to achieve weight maintenance. Qualitative methods may improve our understanding of patient perceptions and so improve treatment for childhood obesity. AIM: To provide insight into the perceptions of parents of obese children as they "journey" from pre-treatment to end of treatment. METHODS: We used purposive sampling and studied 17 parents of children (mean (SD) age 8.4 (2.1) years) attending 6-month outpatient treatments for obesity (BMI>98th percentile). Parent's perceptions were explored by in-depth interviews, analysed using Framework methods. RESULTS: Parents were characterised as being unaware of their child's weight, in denial or actively seeking treatment. Parents were consistently motivated to enter treatment due to perceived benefits to their child's self-esteem or quality of life, and weight outcomes appeared typically less important. During treatment parents felt there was a lack of support for lifestyle changes outside the clinic, and noted that members of the extended family often undermined or failed to support lifestyle changes. Parents generally felt that treatment should have continued beyond 6 months and that it had provided benefits to their child's well-being, self-esteem and quality of life, and this is what motivated many to remain engaged with treatment. DISCUSSION: This study may help inform future treatments for childhood obesity by providing insights into the aspects of treatment of greatest importance to parents. Future treatments may need to consider providing greater support for lifestyle changes within the extended family, and may need to focus more on psycho-social outcomes. PMID- 17916588 TI - The implications of health literacy on patient-provider communication. AB - Limited health literacy has been associated with a range of adverse health outcomes including decreased use of preventive health services, poorer disease specific outcomes for certain chronic conditions and increased risk of hospitalisation and mortality. Although the majority of research has been conducted in the adult population, there is a small and growing body of research on this subject in the paediatric literature. In this article, we will review the research on health literacy, consider the range of other communication skills associated with limited health literacy and explore strategies to improve patient provider communication for clinicians who care for families with limited health literacy skills. PMID- 17916587 TI - Infant morbidity in an Indian slum birth cohort. AB - OBJECTIVE: To establish incidence rates, clinic referrals, hospitalisations, mortality rates and baseline determinants of morbidity among infants in an Indian slum. DESIGN: A community-based birth cohort with twice-weekly surveillance. SETTING: Vellore, South India. SUBJECTS: 452 newborns recruited over 18 months, followed through infancy. MAIN OUTCOME MEASURES: Incidence rates of gastrointestinal illness, respiratory illness, undifferentiated fever, other infections and non-infectious morbidity; rates of community-based diagnoses, clinic visits and hospitalisation; and rate ratios of baseline factors for morbidity. RESULTS: Infants experienced 12 episodes (95% confidence interval (CI) 11 to 13) of illness, spending about one fifth of their infancy with an illness. Respiratory and gastrointestinal symptoms were most common with incidence rates (95% CI) of 7.4 (6.9 to 7.9) and 3.6 (3.3 to 3.9) episodes per child-year. Factors independently associated with a higher incidence of respiratory and gastrointestinal illness were age (3-5 months), male sex, cold/wet season and household involved in beedi work. The rate (95% CI) of hospitalisation, mainly for respiratory and gastrointestinal illness, was 0.28 (0.22 to 0.35) per child year. CONCLUSIONS: The morbidity burden due to respiratory and gastrointestinal illness is high in a South Indian urban slum, with children ill for approximately one fifth of infancy, mainly with respiratory and gastrointestinal illnesses. The risk factors identified were younger age, male sex, cold/wet season and household involvement in beedi work. PMID- 17916589 TI - Is reflux nephropathy preventable, and will the NICE childhood UTI guidelines help? PMID- 17916590 TI - Are there strategies to reduce the length of stay for well near-term babies? PMID- 17916591 TI - Integral protein microarrays for the identification of lung cancer antigens in sera that induce a humoral immune response. AB - The identification of biomarkers (both molecules and profiles) in patient sera offers enormous interest for the diagnosis of cancers. In this context, the detection of antibodies to tumor cell autologous antigens possesses great potential. The humoral immune response represents a form of biological amplification of signals that are otherwise weak because of very low concentrations of antigen, especially in the early stages of cancers. Herein we present the use of integral microarrays spotted with tumor-derived proteins to investigate the antibody repertoire in the sera of lung cancer patients and controls. The use of two-dimensional liquid chromatography allowed us to separate proteins from the lung adenocarcinoma cell line A549 into 1760 fractions, which were printed in duplicate, along with various controls, onto nitrocellulose coated slides. The sensitivity and specificity of the microarrays to detect singular antibodies in fluids were first validated through the recognition of fractions containing a lung marker antigen by antibody probing. Twenty fractions were initially selected as highly reactive against the anti-PGP9.5 antibody, and subsequent mass spectrometry analyses confirmed the identity of PGP9.5 protein in four of them. As a result, the importance of neighboring fractions in microarray detection was revealed due to the spreading of proteins during the separation process. Next, the microarrays were individually incubated with 14 serum samples from patients with lung cancer patients, 14 sera from colon cancer patients, and 14 control sera from normal subjects. The reactivity of the selected fractions was analyzed, and the level of immunoglobulin bound to each fraction by each serum sample was quantified. Eight of the 20 fractions offered p values < 0.01 and were recognized by an average of four reacting patients, whereas no serum from normal individuals was positive for those fractions. Protein microarrays from tumor-derived fractions hold the diagnostic potential of uncovering antigens that induce an immune response in patients with certain types of cancers. PMID- 17916592 TI - Biobanks: transnational, European and global networks. AB - Biobanks contain biological samples and associated information that are essential raw materials for advancement of biotechnology, human health, and research and development in life sciences. Population-based and disease-oriented biobanks are major biobank formats to establish the disease relevance of human genes and provide opportunities to elucidate their interaction with environment and lifestyle. The developments in personalized medicine require molecular definition of new disease subentities and biomarkers for identification of relevant patient subgroups for drug development. These emerging demands can only be met if biobanks cooperate at the transnational or even global scale. Establishment of common standards and strategies to cope with the heterogeneous legal and ethical landscape in different countries are seen as major challenges for biobank networks. The Central Research Infrastructure for Molecular Pathology (CRIP), the concept for a pan-European Biobanking and Biomolecular Resources Research Infrastructure (BBMRI), and the Organization for Economic Co-operation and Development (OECD) global Biological Resources Centres network are examples for transnational, European and global biobank networks that are described in this article. PMID- 17916593 TI - Geographically based investigation of the influence of very-preterm births on routine mortality statistics from the UK and Australia. AB - BACKGROUND: Comparisons of national perinatal and neonatal mortality often neglect the underlying causes. OBJECTIVE: To assess effects of very-preterm births in the UK and Australia. SETTING: Two geographically defined populations: the former Trent Health Region of the UK and New South Wales (NSW)/the Australian Capital Territory (ACT), Australia. METHOD: All births 22(+0) to 31(+6) weeks in 2000, 2001 and 2002 were identified by established surveys of perinatal care. Rates of birth and death were compared. RESULTS: The population of NSW/ACT was 35% higher and there were 66% more births than in Trent (273 495 vs 164 824). The proportion of liveborn infants between 22 and 31 weeks gestation was about 25% higher in Trent (NSW/ACT 2945, rate per 1000 live births 10.82 (95% CI 10.43 to 11.22); Trent 2208, rate per 1000 live births 13.47 (95% CI 12.92 to 14.05)). The proportion of these infants admitted to a neonatal unit was also higher in Trent (91.2% vs 94.4%; OR 1.63 (95% CI 1.30 to 2.05)). Unadjusted mortality in infants admitted to a neonatal unit was similar: NSW/ACT 332/2686 (12.4%); Trent 284/2085 (13.6%); unadjusted OR 1.12 (95% CI 0.94 to 1.33; p = 0.21). CONCLUSIONS: The higher rates of very premature birth and more ready admission to neonatal intensive care for infants in the UK may help to explain why perinatal and neonatal mortality are higher there than in Australia. Efforts to understand why the rate of premature birth in the UK is so high should be a national priority. PMID- 17916594 TI - A cohort study of low Apgar scores and cognitive outcomes. AB - OBJECTIVE: To investigate the association of brief (0-5 minutes) and prolonged (>5 minutes) low Apgar scores (<7) in non-encephalopathic infants with educational achievement at age 15-16 and intelligence quotients (IQs) at age 18. DESIGN: Population-based record-linkage cohort study of 176 524 male infants born throughout Sweden between 1973 and 1976. PATIENTS AND METHODS: Data from the Medical Birth Register were linked to Population and Housing Censuses, conscription medical records (IQ), and school registers (summary school grade). Infants were classified according to the time for their Apgar score to reach 7 or above. Premature infants and those with encephalopathy were excluded. RESULTS: Infants with brief (OR = 1.14 (1.03-1.27)) or prolonged (OR = 1.35 (1.07-1.69)) low Apgar scores were more likely to have a low IQ score. There was an increased risk of a low IQ score (p = 0.003) the longer it took the infant to achieve a normal Apgar score. There was no association between brief (OR = 0.96 (0.87 1.06)) or prolonged (OR = 1.01 (0.81-1.26)) low Apgar scores and a low summary school grade at age 15-16, or evidence for a trend in the risk of a low school grade (p = 0.61). The estimated proportion with an IQ score below 81 due to transiently low Apgar scores was only 0.7%. CONCLUSIONS: Infants in poor condition at birth have increased risk of poor functioning in cognitive tests in later life. This supports the idea of a "continuum of reproductive casualty", although the small individual effect suggests that these mild degrees of fetal compromise are not of clinical importance. PMID- 17916595 TI - Neuroendocrine and inflammatory factors associated with positive affect in healthy men and women: the Whitehall II study. AB - Positive affective states are associated with favorable health outcomes, but the underlying mechanisms are poorly understood. The authors assessed associations between positive affect, cortisol sampled over the day, and inflammatory markers (C-reactive protein and interleukin-6) among 2,873 healthy members of the Whitehall II study. Data for this study were collected in 2002-2004 in London, United Kingdom. Saliva free cortisol was assessed on waking, 30 minutes later, and four times over the day and evening. Positive affect was indexed by aggregating ecological momentary assessments of positive mood over the day. Salivary cortisol averaged over the day was inversely associated with positive affect after controlling for age, gender, income, ethnicity, body mass index, waist/hip ratio, smoking, paid employment, time of waking in the morning, and depression (p = 0.003). There was no association with cortisol responses to waking. The adjusted odds of C-reactive protein >/=3.00 mg/liter was 1.89 (95% confidence interval: 1.08, 3.31) in low- compared with high-positive-affect women, and plasma interleukin-6 was also inversely related to positive affect in women (p = 0.016). Neither inflammatory marker was related to positive affect in men. These results confirm findings from smaller studies relating cortisol with positive affect while suggesting that in women, positive affect is associated with reduced levels of inflammatory markers. PMID- 17916596 TI - Stromal fibroblasts in colorectal liver metastases originate from resident fibroblasts and generate an inflammatory microenvironment. AB - Cancer-associated stromal fibroblasts (CAFs) are the main cellular constituents of reactive stroma in primary and metastatic cancer. We analyzed phenotypical characteristics of CAFs from human colorectal liver metastases (CLMs) and their role in inflammation and cancer progression. CAFs displayed a vimentin(+), alpha smooth-muscle actin(+), and Thy-1(+) phenotype similar to resident portal-located liver fibroblasts (LFs). We demonstrated that CLMs are inflammatory sites showing stromal expression of interleukin-8 (IL-8), a chemokine related to invasion and angiogenesis. In vitro analyses revealed a striking induction of IL-8 expression in CAFs and LFs by tumor necrosis factor-alpha (TNF-alpha). The effect of TNF alpha on CAFs is inhibited by the nuclear factor-kappaB inhibitor parthenolide. Conditioned medium of CAFs and LFs similarly stimulated the migration of DLD-1, Colo-678, HuH7 carcinoma cells, and human umbilical vein endothelial cells in vitro. Pretreatment of CAFs with TNF-alpha increased the chemotaxis of Colo-678 colon carcinoma cells by conditioned medium of CAFs; however, blockage of IL-8 activity showed no inhibitory effect. In conclusion, these data raise the possibility that the majority of CAFs in CLM originate from resident LFs. TNF alpha-induced up-regulation of IL-8 via nuclear factor-kappaB in CAFs is an inflammatory pathway, potentially permissive for cancer invasion that may represent a novel therapeutic target. PMID- 17916597 TI - Snai2 expression enhances ultraviolet radiation-induced skin carcinogenesis. AB - Snai2, encoded by the SNAI2 gene, has been shown to modulate epithelial mesenchymal transformation (EMT), the conversion of sessile epithelial cells attached to adjacent cells and to the basement membrane into dissociated and motile fibroblastic cells. EMT occurs during development, wound healing, and carcinoma progression. Using Snai2-null mice (Snai2(lacZ)), we evaluated the role of Snai2 in UV radiation (UVR)-induced skin carcinogenesis. In chronically UVR exposed nontumor-bearing skin from Snai2-null mice, inflammation and epidermal proliferation were decreased compared with wild-type (+/+) skin. Snai2-null mice had a consistently lower tumor burden than +/+ mice. In addition, null mice developed fewer aggressive spindle cell tumors, believed to arise from squamous cell carcinomas that have undergone EMT, than +/+ mice; however, the difference in tumor type distribution between the two genotypes was not statistically significant. No metastases were observed in either the +/+ or Snai2-null mice. Using quantitative reverse transcriptase-polymerase chain reaction and immunohistochemistry, we showed that the spindle cell tumors in the Snai2-null mice demonstrated impaired EMT, as shown by decreased vimentin and increased cadherin 1 expression. This study confirms a role for Snai2 in EMT, but demonstrates that Snai2 expression is not required for the development or progression of UVR-induced skin tumors. PMID- 17916598 TI - Caveolin-1 is critical for the maturation of tumor blood vessels through the regulation of both endothelial tube formation and mural cell recruitment. AB - In the normal microvasculature, caveolin-1, the structural protein of caveolae, modulates transcytosis and paracellular permeability. Here, we used caveolin-1 deficient mice (Cav(-/-)) to track the potential active roles of caveolin-1 down modulation in the regulation of vascular permeability and morphogenesis in tumors. In B16 melanoma-bearing Cav(-/-) mice, we found that fibrinogen accumulated in early-stage tumors to a larger extent than in wild-type animals. These results were confirmed by the observations of a net elevation of the interstitial fluid pressure and a relative deficit in albumin extravasation in Cav(-/-) tumors (versus healthy tissues). Immunostaining analyses of Cav(-/-) tumor sections further revealed a higher density of CD31-positive vascular structures and a dramatic deficit in alpha-smooth muscle actin-stained mural cells. The increase in blood plasma volume in Cav(-/-) tumors was confirmed by dynamic contrast enhanced-magnetic resonance imaging and found to be associated with a more rapid tumor growth. Finally, an in vitro wound test and the aorta ring assay revealed that silencing caveolin expression could directly impair the migration and the outgrowth of smooth muscle cells/pericytes, particularly in response to platelet-derived growth factor. In conclusion, a decrease in caveolin abundance, by promoting angiogenesis and preventing its termination by mural cell recruitment, appears as an important control point for the formation of new tumor blood vessels. Caveolin-1 therefore has the potential to be a marker of tumor vasculature maturity that may help adjusting anticancer therapies. PMID- 17916599 TI - Increased Goodpasture antigen-binding protein expression induces type IV collagen disorganization and deposit of immunoglobulin A in glomerular basement membrane. AB - Increased expression of Goodpasture antigen-binding protein (GPBP), a protein that binds and phosphorylates basement membrane collagen, has been associated with immune complex-mediated pathogenesis. However, recent reports have questioned this biological function and proposed that GPBP serves as a cytosolic ceramide transporter (CERT(L)). Thus, the role of GPBP in vivo remains unknown. New Zealand White (NZW) mice are considered healthy animals although they convey a genetic predisposition for immune complex-mediated glomerulonephritis. Here we show that NZW mice developed age-dependent lupus-prone autoimmune response and immune complex-mediated glomerulonephritis characterized by elevated GPBP, glomerular basement membrane (GBM) collagen disorganization and expansion, and deposits of IgA on disrupted GBM. Transgenic overexpression of human GPBP (hGPBP) in non-lupus-prone mice triggered similar glomerular abnormalities including deposits of IgA on a capillary GBM that underwent dissociation, in the absence of an evident autoimmune response. We provide in vivo evidence that GPBP regulates GBM collagen organization and its elevated expression causes dissociation and subsequent accumulation of IgA on the GBM. Finally, we describe a previously unrecognized pathogenic mechanism that may be relevant in human primary immune complex-mediated glomerulonephritis. PMID- 17916600 TI - Multiplex PCR detection of GSTM1, GSTT1, and GSTP1 gene variants: simultaneously detecting GSTM1 and GSTT1 gene copy number and the allelic status of the GSTP1 Ile105Val genetic variant. AB - The glutathione S-transferase (GST) genes GSTM1, GSTT1, and GSTP1 are involved in the detoxification of a broad range of toxic substances. Genetic polymorphisms in these genes have been studied intensively for their potential role in cancer susceptibility and drug response. In Caucasians, the enzyme activity of GSTM1 and GSTT1 is absent in approximately 50 and 15% of the population, respectively, due to deletions of both chromosomal copies of the genes. A trimodal phenotype pattern exists in which individuals with two, one, or no functional genes are fast, intermediate, or slow "conjugators," respectively. Most studies investigating the effect of the GSTM1 and GSTT1 deletions do not distinguish between fast and intermediate conjugators because the applied genotyping assays only detect if at least one copy of either gene is present. We present a multiplex PCR assay that detects if an individual has none, one, or two copies of the GSTM1 and GSTT1 genes and simultaneously detects the allelic status of the GSTP1 Ile105Val genetic variant. A total of 200 Danes, 100 Somalis, and 100 Greenlanders were genotyped. This multiplex PCR assay enables future large-scale studies to investigate the role of GSTs. PMID- 17916601 TI - A molecular fraction collecting tool for the ABI 310 automated sequencer. AB - Several methods exist to retrieve and purify DNA fragments after agarose or polyacrylamide gel electrophoresis for subsequent analyses. However, molecules present in low concentration and molecules similar in size to their neighbors are difficult to purify. Capillary electrophoresis has become popular in molecular diagnostic laboratories because of its automation, excellent resolution, and high sensitivity. In the current study, the ABI Prism 310 Genetic Analyzer was reconfigured into a fraction collector by adapting the standard gel block to accommodate a collection tube at the distal end of capillary. The time to collect the desired peaks was estimated by extrapolating from standard capillary electrophoresis using the original gel block. Fraction collection from a mixture of DNA fragments amplified from wild type and several internal tandem duplication mutations of the FMS-like tyrosine kinase 3 (Flt3) gene yielded highly purified DNA fragments containing internal tandem duplication mutations and predictable electrokinetics using the reconstructed gel block. The reconfigured instrument could successfully isolate DNA amplicons from extremely low-amplitude peaks (110 relative fluorescent units), which were undetectable using polyacrylamide gel electrophoresis. In addition, we successfully isolated bands that were only three bases apart that comigrated on polyacrylamide gel electrophoresis. DNA sequencing was used to confirm that the correct peaks were recovered at sufficient purity. PMID- 17916602 TI - A new dimethyl sulfoxide-based method for gene promoter methylation detection. AB - The identification of gene promoter methylation is a useful tool for the molecular diagnosis of human diseases. We have developed a new PCR-based technique for detecting the methylation status of CpG islands of gene promoters. This new method, named methyl-sensitive dimethyl sulfoxide-PCR (Ms-DMSO-PCR), is based on the finding that methylated and unmethylated DNAs show a different sensitivity to the amount of DMSO used in the PCR reaction. For the amplification of methylated DNA, more DMSO is required in comparison to unmethylated DNA. This finding resulted in the development of a simple PCR screening of CpG islands with addition of DMSO in the range from 0 to 8% (v/v), and the same pair of primers is sufficient for distinguishing hyper- or hypomethylated gene promoters from normally methylated sequences. This new technique is a one-step procedure and does not require any modifications of DNA or expensive equipment. Therefore, Ms DMSO-PCR has the potential to be widely used for clinical applications as well in basic research. PMID- 17916603 TI - Is occult lymph node disease in colorectal cancer patients clinically significant? A review of the relevant literature. AB - The clinical significance of micrometastasis of colorectal cancer (CRC) to regional lymph nodes remains controversial. In this review, we analyze publications that have evaluated the clinical significance of occult lymph node metastasis in CRC. An extensive literature search identified 19 publications that evaluated the clinical significance of micrometastatic CRC by various methods, including immunohistochemistry (IHC; n = 13) and reverse transcription-polymerase chain reaction (RT-PCR, n = 6). These studies were reviewed for methodology and findings. Significant limitations in methodology were identified, including inconsistent histological definitions of micrometastatic disease, poor sampling because of an inadequate number of lymph nodes or number of sections per lymph node analyzed, lack of conformity with respect to IHC antibody or RT-PCR marker, and inadequate power because of small sample size. Micrometastatic lymph node metastasis identified by RT-PCR was consistently found to be prognostically significant, but this was not true of micrometastatic disease identified by IHC. RT-PCR analysis of lymph nodes with specific markers can help identify pN0 (pathological-negative lymph node) CRC patients at increased risk for recurrence. The identification of occult disease by IHC techniques may also ultimately prove to be associated with worse outcome, but a number of inadequately powered studies have concluded conversely. PMID- 17916604 TI - Time to give nutrition interventions a higher profile: cost-effectiveness of 10 nutrition interventions. AB - The aim of this study was to evaluate the economic performance of 10 nutrition interventions. The interventions included Mediterranean Diet, Intensive Lifestyle Change (nutrition and physical activity) to Prevent Diabetes, Reduced Fat Diet for persons with IGT, Nutritional Counselling in GP (GP, general practice/primary care), Nurse Counselling in GP, Oxcheck Nurse Health Checks in GP, Gutbusters Workplace (for men), Talking Computer, Multi Media 2 fruit 5 veg Campaign and the FFFF (Fighting Fit, Fighting Fat) Media Campaign. Markov models were constructed in order to estimate economic performance expressed as cost per QALY (quality adjusted life year) gained. Data from original clinical trial reports were used to populate the models, supplemented by the wider literature where required. Performance of the Mediterranean Diet and Intensive Lifestyle Change to Prevent Diabetes interventions could be estimated with most certainty and both were highly cost-effective interventions, at AU $1020 (US $760, 410 pounds) and AU $1880 (US $1410, 750 pounds) per QALY gained, respectively. The media campaign interventions appear highly cost-effective at AU $46 (US $34, 18 pounds) for '2 fruit 5 veg' and AU $5600 (US $4200, 2200 pounds) per QALY gained for FFFF, but are associated with considerable uncertainty, and may be dominated under certain assumptions. Several interventions were cost-saving under plausible sets of assumptions, whereas a small number were potentially dominated. All interventions subject to economic evaluation appeared cost-effective relative to societal norms. Nutrition interventions can constitute a highly efficient component of a strategy to reduce the growing disease burden linked to over/poor nutrition. There is an urgent need for high-quality trial data from which economic performance of nutrition interventions can be modelled. PMID- 17916605 TI - Insights into the role of the respiratory muscle metaboreflex. PMID- 17916606 TI - pH dependence of melanoma cell migration: protons extruded by NHE1 dominate protons of the bulk solution. AB - Migration and morphology of human melanoma cells (MV3) depend on extracellular pH (pHe) and the activity of the Na+/H+ exchanger NHE1. To distinguish effects of NHE1 activity per se from effects of pHe we compared an NHE1-deficient mutant with rescued and wild-type cells. Time lapse video microscopy was used to investigate migratory and morphological effects caused by pHe and NHE1 activity, and a membrane-bound fluorescein conjugate was employed for ratiometric pH measurements at the outer leaflet of the cell membrane. As long as NHE1 remained inactive due to deficiency or inhibition by cariporide (HOE642) neither migration nor morphology was affected by changes in pHe. Under these conditions pH at the outer leaflet of the plasma membrane was uniform all over the cell surface. The typical pH dependence of MV3 cell migration and morphology could be reconstituted by restoring NHE1 activity. At the same time the proton gradient at the outer leaflet of the plasma membrane with the higher proton concentration at the leading edge and the lower one at the cell rear was re-established as well. Hence, NHE1 activity generates a proton gradient at the cell surface accompanied by the cells' ability to respond to changes in pHe (bulk pH). We conclude that NHE1 activity contributes to the generation of a well-defined cell surface pH by creating a proton gradient at the outer leaflet of the plasma membrane that is needed for (i) the development of a variety of morphologies including a distinct polarity and (ii) migration. A missing proton gradient at the cell surface cannot be compensated for by varying pHe. PMID- 17916608 TI - Synaptic heterogeneity between mouse paracapsular intercalated neurons of the amygdala. AB - GABAergic medial paracapsular intercalated (Imp) neurons of amygdala are thought of as playing a central role in fear learning and extinction. We report here that the synaptic network formed by these neurons exhibits distinct short-term plastic synaptic responses. The success rate of synaptic events evoked at a frequency range of 0.1-10 Hz varied dramatically between different connected cell pairs. Upon enhancing the frequency of stimulation, the success rate increased, decreased or remained constant, in a similar number of cell pairs. Such synaptic heterogeneity resulted in inhibition of the firing of the postsynaptic neurons with different efficacies. Moreover, we found that the different synaptic weights were mainly determined by diversity in presynaptic release probabilities rather than postsynaptic changes. Sequential paired recording experiments demonstrated that the same presynaptic neuron established the same type of synaptic connections with different postsynaptic neurons, suggesting the absence of target cell specificity. Conversely, the same postsynaptic neuron was contacted by different types of synaptic connections formed by different presynaptic neurons. A detailed anatomical analysis of the recorded neurons revealed discrete and unexpected peculiarities in the dendritic and axonal patterns of different cell pairs. In contrast, several intrinsic electrophysiological responses were homogeneous among neurons, and synaptic failure counts were not affected by presynaptic cannabinoid 1 or GABA B receptors. We propose that the heterogeneous functional connectivity of Imp neurons, demonstrated by this study, is required to maintain the stability of firing patterns which is critical for the computational role of the amygdala in fear learning and extinction. PMID- 17916607 TI - Increased Ca2+ influx through Na+/Ca2+ exchanger during long-term facilitation at crayfish neuromuscular junctions. AB - Intense motor neuron activity induces a long-term facilitation (LTF) of synaptic transmission at crayfish neuromuscular junctions (NMJs) that is accompanied by an increase in the accumulation of presynaptic Ca2+ ions during a test train of action potentials. It is natural to assume that the increased Ca2+ influx during action potentials is directly responsible for the increased transmitter release in LTF, especially as the magnitudes of LTF and increased Ca2+ influx are positively correlated. However, our results indicate that the elevated Ca2+ entry occurs through the reverse mode operation of presynaptic Na+/Ca2+ exchangers that are activated by an LTF-inducing tetanus. Inhibition of Na+/Ca2+ exchange blocks this additional Ca2+ influx without affecting LTF, showing that LTF is not a consequence of the regulation of these transporters and is not directly related to the increase in [Ca2+]i reached during a train of action potentials. Their correlation is probably due to both being induced independently by the strong [Ca2+]i elevation accompanying LTF-inducing stimuli. Our results reveal a new form of regulation of neuronal Na+/Ca2+ exchange that does not directly alter the strength of synaptic transmission. PMID- 17916609 TI - Force generation examined by laser temperature-jumps in shortening and lengthening mammalian (rabbit psoas) muscle fibres. AB - We examined the tension change induced by a rapid temperature jump (T-jump) in shortening and lengthening active muscle fibres. Experiments were done on segments of permeabilized single fibres (length (L0) approximately 2 mm, sarcomere length 2.5 microm) from rabbit psoas muscle; [MgATP] was 4.6 mm, pH 7.1, ionic strength 200 mm and temperature approximately 9 degrees C. A fibre was maximally Ca2+-activated in the isometric state and a approximately 3 degrees C, rapid (< 0.2 ms), laser T-jump applied when the tension was approximately steady in the isometric state, or during ramp shortening or ramp lengthening at a limited range of velocities (0-0.2 L0 s(-1)). The tension increased to 2- to 3 x P0 (isometric force) during ramp lengthening at velocities > 0.05 L0 s(-1), whereas the tension decreased to about < 0.5 x P0 during shortening at 0.1-0.2 L0 s(-1); the unloaded shortening velocity was approximately 1 L0 s(-1) and the curvature of the force-shortening velocity relation was high (a/P0 ratio from Hill's equation of approximately 0.05). In isometric state, a T-jump induced a tension rise of 15-20% to a new steady state; by curve fitting, the tension rise could be resolved into a fast (phase 2b, 40-50 s(-1)) and a slow (phase 3, 5-10 s(-1)) exponential component (as previously reported). During steady lengthening, a T-jump induced a small instantaneous drop in tension, followed by recovery, so that the final tension recorded with and without a T-jump was not significantly different; thus, a T-jump did not lead to a net increase of tension. During steady shortening, the T-jump induced a pronounced tension rise and both its amplitude and the rate (from a single exponential fit) increased with shortening velocity; at 0.1-0.2 L0 s(-1), the extent of fibre shortening during the T-jump tension rise was estimated to be approximately 1.2% L(0) and it was shorter at lower velocities. At a given shortening velocity and over the temperature range of 8-30 degrees C, the rate of T-jump tension rise increased with warming (Q10 approximately 2.7), similar to phase 2b (endothermic force generation) in isometric muscle. Results are discussed in relation to the previous findings in isometric muscle fibres which showed that a T-jump promotes an early step in the crossbridge-ATPase cycle that generates force. In general, the finding that the T jump effect on active muscle tension is pronounced during shortening, but is depressed/inhibited during lengthening, is consistent with the expectations from the Fenn effect that energy liberation (and acto-myosin ATPase rate) in muscle are increased during shortening and depressed/inhibited during lengthening. PMID- 17916610 TI - Postnatal development of synaptic transmission in local networks of L5A pyramidal neurons in rat somatosensory cortex. AB - The probability of synaptic transmitter release determines the spread of excitation and the possible range of computations at unitary connections. To investigate whether synaptic properties between neocortical pyramidal neurons change during the assembly period of cortical circuits, whole-cell voltage recordings were made simultaneously from two layer 5A (L5A) pyramidal neurons within the cortical columns of rat barrel cortex. We found that synaptic transmission between L5A pyramidal neurons is very reliable between 2 and 3 weeks of postnatal development with a mean unitary EPSP amplitude of approximately 1.2 mV, but becomes less efficient and fails more frequently in the more mature cortex of approximately 4 weeks of age with a mean unitary EPSP amplitude of 0.65 mV. Coefficient of variation and failure rate increase as the unitary EPSP amplitude decreases during development. The paired-pulse ratio (PPR) of synaptic efficacy at 10 Hz changes from 0.7 to 1.04. Despite the overall increase in PPR, short-term plasticity displays a large variability at 4 weeks, ranging from strong depression to strong facilitation (PPR, range 0.6-2.1), suggesting the potential for use-dependent modifications at this intracortical synapse. In conclusion, the transmitter release probability at the L5A-L5A connection is developmentally regulated in such a way that in juvenile animals excitation by single action potentials is efficiently transmitted, whereas in the more mature cortex synapses might be endowed with a diversity of filtering characteristics. PMID- 17916611 TI - NOS isoform-specific regulation of basal but not exercise-induced mitochondrial biogenesis in mouse skeletal muscle. AB - Nitric oxide is a potential regulator of mitochondrial biogenesis. Therefore, we investigated if mice deficient in endothelial nitric oxide synthase (eNOS-/-) or neuronal NOS (nNOS-/-) have attenuated activation of skeletal muscle mitochondrial biogenesis in response to exercise. eNOS-/-, nNOS-/- and C57Bl/6 (CON) mice (16.3 +/- 0.2 weeks old) either remained in their cages (basal) or ran on a treadmill (16 m min(-1), 5% grade) for 60 min (n = 8 per group) and were killed 6 h after exercise. Other eNOS-/-, nNOS-/- and CON mice exercise trained for 9 days (60 min per day) and were killed 24 h after the last bout of exercise training. eNOS-/- mice had significantly higher nNOS protein and nNOS-/- mice had significantly higher eNOS protein in the EDL, but not the soleus. The basal mitochondrial biogenesis markers NRF1, NRF2alpha and mtTFA mRNA were significantly (P< 0.05) higher in the soleus and EDL of nNOS-/- mice whilst basal citrate synthase activity was higher in the soleus and basal PGC-1alpha mRNA higher in the EDL. Also, eNOS-/- mice had significantly higher basal citrate synthase activity in the soleus but not the EDL. Acute exercise increased (P< 0.05) PGC-1alpha mRNA in soleus and EDL and NRF2alpha mRNA in the EDL to a similar extent in all genotypes. In addition, short-term exercise training significantly increased cytochrome c protein in all genotypes (P< 0.05) in the EDL. In conclusion, eNOS and nNOS are differentially involved in the basal regulation of mitochondrial biogenesis in skeletal muscle but are not critical for exercise-induced increases in mitochondrial biogenesis in skeletal muscle. PMID- 17916612 TI - Antioxidant administration attenuates mechanical ventilation-induced rat diaphragm muscle atrophy independent of protein kinase B (PKB Akt) signalling. AB - Oxidative stress promotes controlled mechanical ventilation (MV)-induced diaphragmatic atrophy. Nonetheless, the signalling pathways responsible for oxidative stress-induced muscle atrophy remain unknown. We tested the hypothesis that oxidative stress down-regulates insulin-like growth factor-1 phosphotidylinositol 3-kinase-protein kinase B serine threonine kinase (IGF-1 PI3K-Akt) signalling and activates the forkhead box O (FoxO) class of transcription factors in diaphragm fibres during MV-induced diaphragm inactivity. Sprague-Dawley rats were randomly assigned to one of five experimental groups: (1) control (Con), (2) 6 h of MV, (3) 6 h of MV with infusion of the antioxidant Trolox, (4) 18 h of MV, (5) 18 h of MV with Trolox. Following 6 h and 18 h of MV, diaphragmatic Akt activation decreased in parallel with increased nuclear localization and transcriptional activation of FoxO1 and decreased nuclear localization of FoxO3 and FoxO4, culminating in increased expression of the muscle-specific ubiquitin ligases, muscle atrophy factor (MAFbx) and muscle ring finger-1 (MuRF-1). Interestingly, following 18 h of MV, antioxidant administration was associated with attenuation of MV-induced atrophy in type I, type IIa and type IIb/IIx myofibres. Collectively, these data reveal that the antioxidant Trolox attenuates MV-induced diaphragmatic atrophy independent of alterations in Akt regulation of FoxO transcription factors and expression of MAFbx or MuRF-1. Further, these results also indicate that differential regulation of diaphragmatic IGF-1-PI3K-Akt signalling exists during the early and late stages of MV. PMID- 17916613 TI - Kinetics of both synchronous and asynchronous quantal release during trains of action potential-evoked EPSCs at the rat calyx of Held. AB - We studied the kinetics of transmitter release during trains of action potential (AP)-evoked excitatory postsynaptic currents (EPSCs) at the calyx of Held synapse of juvenile rats. Using a new quantitative method based on a combination of ensemble fluctuation analysis and deconvolution, we were able to analyse mean quantal size (q) and release rate (xi) continuously in a time-resolved manner. Estimates derived this way agreed well with values of q and quantal content (M) calculated for each EPSC within the train from ensemble means of peak amplitudes and their variances. Separate analysis of synchronous and asynchronous quantal release during long stimulus trains (200 ms, 100 Hz) revealed that the latter component was highly variable among different synapses but it was unequivocally identified in 18 out of 37 synapses analysed. Peak rates of asynchronous release ranged from 0.2 to 15.2 vesicles ms(-1) (ves ms(-1)) with a mean of 2.3 +/- 0.6 ves ms(-1). On average, asynchronous release accounted for less than 14% of the total number of about 3670 +/- 350 vesicles released during 200 ms trains. Following such trains, asynchronous release decayed with several time constants, the fastest one being in the order of 15 ms. The short duration of asynchronous release at the calyx of Held synapse may aid in generating brief postsynaptic depolarizations, avoiding temporal summation and preserving action potential timing during high frequency bursts. PMID- 17916614 TI - Properties of the major classes of mechanoreceptors in the guinea pig bladder. AB - Sensory neurons represent an attractive target for pharmacological treatment of various bladder disorders. However the properties of major classes of mechano sensory neurons projecting to the bladder have not been systematically established. An in vitro bladder preparation was used to examine the effects of a range of mechanical stimuli (stretch, von Frey hair stroking and focal compression of receptive fields) and chemical stimuli (1 mm alpha,beta-methylene ATP, hypertonic solutions (500 mm NaCl) and 3 microm capsaicin) during electrophysiological recordings from guinea pig bladder afferents. Four functionally distinct populations of bladder sensory neurons were distinguished by these stimuli. The first class, muscle mechanoreceptors, were activated by stretch but not by mucosal stroking with light (0.05-0.1 mN) von Frey hairs or by hypertonic saline, alpha,beta-methylene ATP or capsaicin. Removal of the urothelium did not affect their stretch-induced firing. The second class, muscle mucosal mechanoreceptors, were activated by both stretch and mucosal stroking with light von Frey hairs or by hypertonic saline and by alpha,beta-methylene ATP, but not by capsaicin. Removal of the urothelium reduced their stretch- and stroking-induced firing. The third class, mucosal high-responding mechanoreceptors, were stretch-insensitive but could be activated by mucosal stroking with light von Frey hairs or by hypertonic saline, alpha,beta-methylene ATP and capsaicin. Stroking-induced firing was significantly reduced by removal of the urothelium. The fourth class, mucosal low-responding mechanoreceptors, were stretch insensitive but could be weakly activated by mucosal stroking with light von Frey hairs but not by hypertonic saline, alpha,beta-methylene ATP or capsaicin. Removal of the urothelium reduced mucosal stroking-induced firing. All four populations of afferents conducted in the C-fibre range and showed class dependent differences in spike amplitude and duration. At least four functional classes of bladder mechanoreceptors can be readily distinguished by different mechanisms of activation and are likely to transmit different types of information to the central nervous system. PMID- 17916616 TI - Abstracts from the 15th Annual European Public Health Association (EUPHA) Conference. October 11-13 2007. Helsinki, Finland. PMID- 17916615 TI - Neural drive to human genioglossus in obstructive sleep apnoea. AB - One postulated mechanism for obstructive sleep apnoea (OSA) is insufficient drive to the upper-airway musculature during sleep, with increased (compensatory) drive during wakefulness. This generates more electromyographic activity in upper airway muscles including genioglossus. To understand drives to upper airway muscles, we recorded single motor unit activity from genioglossus in male groups of control (n = 7, 7 +/- 2 events h(-1)) and severe OSA (n = 9, 54 +/- 4 events h(-1)) subjects. One hundred and seventy-eight genioglossus units were recorded using monopolar electrodes. Subjects were awake, supine and breathing through a nasal mask. The distribution of the six types of motor unit activity in genioglossus (Inspiratory Phasic, Inspiratory Tonic, Expiratory Phasic, Expiratory Tonic, Tonic and Tonic Other) was identical in both groups. Single unit action potentials in OSA were larger in area (by 34%, P < 0.05) and longer in duration (by 23%, P < 0.05). Inspiratory units were recruited earlier in OSA than control subjects. In control subjects, Inspiratory Tonic units peaked earlier than Inspiratory Phasic units, while in OSA subjects, Inspiratory Tonic and Phasic units peaked simultaneously. Onset frequencies did not differ between groups, but the peak discharge frequency for Inspiratory Phasic units was higher in OSA (22 +/- 1 Hz) than control subjects (19 +/- 1 Hz, P = 0.003), but conversely, the peak discharge frequency of Inspiratory Tonic units was higher in control subjects (28 +/- 1 Hz versus 25 +/- 1 Hz, P < 0.05). Increased motor unit action potential area indicates that neurogenic changes have occurred in OSA. In addition, the differences in the timing and firing frequency of the inspiratory classes of genioglossus motor units indicate that the output of the hypoglossal nucleus may have changed. PMID- 17916623 TI - Locust plagues, climate variation, and the rhythms of nature. PMID- 17916625 TI - Genetic analysis reveals functional redundancy and the major target genes of the Arabidopsis miR159 family. AB - Currently, there are very few loss-of-function mutations in micro-RNA genes. Here, we characterize two members of the Arabidopsis MIR159 family, miR159a and miR159b, that are predicted to regulate the expression of a family of seven transcription factors that includes the two redundant GAMYB-like genes, MYB33 and MYB65. Using transfer DNA (T-DNA) insertional mutants, we show that a mir159ab double mutant has pleiotropic morphological defects, including altered growth habit, curled leaves, small siliques, and small seeds. Neither mir159a nor mir159b single mutants displayed any of these traits, indicating functional redundancy. By using reporter-gene constructs, it appears that MIR159a and MIR159b are transcribed almost exclusively in the cells in which MYB33 is repressed, as had been previously determined by comparison of MYB33 and mMYB33 (an miR159-resistant allele of MYB33) expression patterns. Consistent with these overlapping transcriptional domains, MYB33 and MYB65 expression levels were elevated throughout mir159ab plants. By contrast, the other five GAMYB-like family members are transcribed predominantly in tissues where miR159a and miR159b are absent, and consequently their expression levels are not markedly elevated in mir159ab. Additionally, mMYB33 transgenic plants can phenocopy the mir159ab phenotype, suggesting that its phenotype is explained by deregulated expression of the redundant gene pair MYB33 and MYB65. This prediction was confirmed; the pleiotropic developmental defects of mir159ab are suppressed through the combined mutations of MYB33 and MYB65, demonstrating the narrow and specific target range of miR159a and miR159b. PMID- 17916624 TI - Neuroglobin dynamics observed with ultrafast 2D-IR vibrational echo spectroscopy. AB - Neuroglobin (Ngb), a protein in the globin family, is found in vertebrate brains. It binds oxygen reversibly. Compared with myoglobin (Mb), the amino acid sequence has limited similarity, but key residues around the heme and the classical globin fold are conserved in Ngb. The CO adduct of Ngb displays two CO absorption bands in the IR spectrum, referred to as N(3) (distal histidine in the pocket) and N(0) (distal histidine swung out of the pocket), which have absorption spectra that are almost identical with the Mb mutants L29F and H64V, respectively. The Mb mutants mimic the heme pocket structures of the corresponding Ngb conformers. The equilibrium protein dynamics for the CO adduct of Ngb are investigated by using ultrafast 2D-IR vibrational echo spectroscopy by observing the CO vibration's spectral diffusion (2D-IR spectra time dependence) and comparing the results with those for the Mb mutants. Although the heme pocket structure and the CO FTIR peak positions of Ngb are similar to those of the mutant Mb proteins, the 2D-IR results demonstrate that the fast structural fluctuations of Ngb are significantly slower than those of the mutant Mbs. The results may also provide some insights into the nature of the energy landscape in the vicinity of the folded protein free energy minimum. PMID- 17916626 TI - Evidence that androgens regulate early developmental events, prior to sexual differentiation. AB - Androgen signaling is critical for normal fetal development but is not thought to regulate events in early embryogenesis. Given the interest in factors controlling the differentiation of embryonic stem (ES) cells, we have explored the possibility that androgens may play a role. This study demonstrates expression of androgen receptor (AR) RNA and protein in four independent mouse ES (mES) cell lines, and shows that the AR is functional and can interact with transfected androgen response elements to promote green fluorescent protein expression. AR mRNA was detected throughout 10-d differentiation in embryoid bodies (EBs). Exposure of EBs to testosterone (T) or dihydrotestosterone, at doses of 1 and 0.1 mum, respectively, promoted formation of beating cardiomyocytes. Flow cytometric analyses demonstrated a significant increase in the number of alpha-actinin and tropomyosin (cardiac markers) positive cells after these treatments. Addition of flutamide (1 microM) to T-treated EBs inhibited the T-induced proliferation of cardiomyocytes, confirming that, in this instance, androgens act via the classical AR-mediated genomic pathway. We also report that mES cells express key steroidogenic enzymes, as detected by RT-PCR, and during 24-h incubations secrete T at concentrations of 1.38 +/- 0.22 nM, levels comparable to those secreted by cultured Leydig cells. These novel data demonstrate the capacity of androgens to stimulate increased differentiation of mouse ES cells to cardiomyocytes, and are in keeping with recent observations that AR-deficient mice exhibit cardiac impairment in adulthood. PMID- 17916627 TI - Gonadotropin-releasing hormone-1 neuronal activity is independent of cyclic nucleotide-gated channels. AB - Pulsatile release of GnRH-1 is essential for secretion of gonadotropin hormones. The frequency of GnRH-1 pulses is regulated during the reproductive cycle by numerous neurotransmitters. Cyclic nucleotide-gated (CNG) channels have been proposed as a mechanism to integrate the cAMP signal evoked by many neurotransmitters. This study reports the expression of the CNGA2 subunit in GnRH 1 neurons obtained from mouse nasal explants and shows the ability of GnRH-1 neurons to increase their activity in response to forskolin (activator of adenylyl cyclases), or 3-isobutyl-1-methylxanthine (inhibitor of phosphodiesterases) even after removal of gamma-aminobutyric acid (A)-ergic input. Next, the endogenous activity of adenylyl cyclases was evaluated as a component of the oscillatory mechanism of GnRH-1 neurons. Inhibition of endogenous activity of adenylyl cyclases did not alter GnRH-1 activity. The potential involvement of CNGA2 subunit in basal or induced activity was tested on GnRH-1 neurons obtained from CNGA2-deficient mice. Without up-regulation of CNGA1 or CNGA3, the absence of functional CNGA2 did not alter either the endogenous GnRH-1 neuronal activity or the response to forskolin, negating CNG channels from cAMP-sensitive mechanisms leading to changes in GnRH-1 neuronal activity. In addition, the potential role of CNGA2 subunit in the synchronization of calcium oscillations previously described was evaluated in GnRH-1 neurons from CNGA2 deficient explants. Synchronized calcium oscillations persisted in CNGA2 deficient GnRH-1 neurons. Taken together, these results indicate that CNGA2 channels are not necessary for either the response of GnRH-1 neurons to cAMP increases or the basal rhythmic activity of GnRH-1 neurons. PMID- 17916630 TI - Human chorionic gonadotropin induces nestin expression in endothelial cells of the ovary via vascular endothelial growth factor signaling. AB - The intermediate filament protein nestin was originally found to be expressed in neuronal progenitor cells, but recent studies have shown that other cell types, including endocrine and vascular endothelial cells, express nestin. In the present study, we examined the expression and localization of nestin in the ovaries of developing, peripubertal, and adult rats. RT-PCR and Western blot analyses revealed that nestin mRNA and proteins were expressed in adult rat ovaries. Immunohistochemical analyses using adult rat ovaries showed that nestin was mainly localized to capillary endothelial cells of theca interna in follicles with more than two layers of granulosa cells and that its expression increased with follicle growth. Ontogenetically, ovarian nestin expression started at the peripubertal period when the first gonadotropin surge occurs. To test the possibility that gonadotropins induce nestin expression, prepubertal (postnatal d 21) rats were sc injected with equine chorionic gonadotropin (eCG) and/or human chorionic gonadotropin (hCG). A single injection of hCG, but not eCG, was sufficient to induce nestin expression in follicles, mainly in capillary endothelial cells of theca interna. Furthermore, pretreatment with an inhibitor of vascular endothelial growth factor receptor prevented the induction of the nestin expression by hCG. These findings demonstrate that the endogenous LH surge induces nestin expression in capillary endothelial cells of theca interna via the vascular endothelial growth factor signaling pathway. Nestin may be involved in angiogenesis in growing follicles, which is followed by follicle maturation and subsequent ovulation. PMID- 17916628 TI - Molecular cloning and characterization of estrogen, androgen, and progesterone nuclear receptors from a freshwater turtle (Pseudemys nelsoni). AB - Steroid hormones are essential for the normal function of many organ systems in vertebrates. Reproductive activity in females and males, such as the differentiation, growth, and maintenance of the reproductive system, requires signaling by the sex steroids. Although extensively studied in mammals and a few fish, amphibians, and bird species, the molecular mechanisms of sex steroid hormone (estrogens, androgens, and progestins) action are poorly understood in reptiles. Here we evaluate hormone receptor ligand interactions in a freshwater turtle, the red-belly slider (Pseudemys nelsoni), after the isolation of cDNAs encoding an estrogen receptor alpha (ERalpha), an androgen receptor (AR), and a progesterone receptor (PR). The full-length red-belly slider turtle (t)ERalpha, tAR, and tPR cDNAs were obtained using 5' and 3' rapid amplification cDNA ends. The deduced amino acid sequences showed high identity to the chicken orthologs (tERalpha, 90%; tAR, 71%; tPR, 71%). Using transient transfection assays of mammalian cells, tERalpha protein displayed estrogen-dependent activation of transcription from an estrogen-responsive element-containing promoter. The other receptor proteins, tAR and tPR, also displayed androgen- or progestin-dependent activation of transcription from androgen- and progestin-responsive murine mammary tumor virus promoters. We further examined the transactivation of tERalpha, tAR and tPR by ligands using a modified GAL4-transactivation system. We found that the GAL4-transactivation system was not suitable for the measurement of tAR and tPR transactivations. This is the first report of the full coding regions of a reptilian AR and PR and the examination of their transactivation by steroid hormones. PMID- 17916629 TI - Insulin receptor substrate-1 deficiency promotes apoptosis in the putative intestinal crypt stem cell region, limits Apcmin/+ tumors, and regulates Sox9. AB - Reduced apoptosis of crypt stem/progenitor cells and elevated insulin and IGFs are linked to colon cancer risk. Insulin receptor substrate-1 (IRS-1) mediates the actions of insulin, IGF-I, and IGF-II, but the role of endogenous IRS-1 in crypt apoptosis and cancer is undefined. Using IRS-1(-/-), IRS-1(+/-), and IRS 1(+/+) mice, we tested the hypothesis that reduced IRS-1 expression increases apoptosis of intestinal crypt cells and protects against Apc(min/+) (Min)/beta catenin-driven intestinal tumors. Expression of Sox9, a transcriptional target of Tcf/beta-catenin and putative biomarker of crypt stem cells, was assessed in intestine of different IRS-1 genotypes and cell lines. Irradiation-induced apoptosis was significantly increased in the crypts and crypt stem cell region of IRS-1-deficient mice. Tumor load was significantly reduced by 31.2 +/- 14.6% in IRS-1(+/-)/Min and by 64.1 +/- 7.6% in IRS-1(-/-)/Min mice, with more prominent reductions in tumor number than size. Compared with IRS-1(+/+)/Min, IRS-1(-/ )/Min mice had fewer Sox9-positive cells in intestinal crypts and reduced Sox9 mRNA in intestine. IRS-1 overexpression increased Sox9 expression in an intestinal epithelial cell line. We conclude that even small reductions in endogenous IRS-1 increase apoptosis of crypt stem or progenitor cells, protect against beta-catenin-driven intestinal tumors, and reduce Sox9, a Tcf/beta catenin target and putative stem/progenitor cell biomarker. PMID- 17916631 TI - Contraction in human myometrium is associated with changes in small heat shock proteins. AB - The myometrium undergoes substantial remodeling at the time of labor including rearrangement of the cellular contractile machinery. The regulation of this process in human myometrium at the time of labor is poorly defined, but evidence in other muscle types suggests modulation by small heat shock proteins (sHSP). The aim of this study was to investigate whether similar changes in sHSP occur in the myometrium at labor. Using a quantitative proteomic approach (two-dimensional difference gel electrophoresis), we found a 69% decrease in the sHSP alphaB crystallin in the myometrium at labor plus multiple isoforms of HSP27. Immunoblotting using phosphospecific HSP27 antibodies (HSP27-serine15, -78, and 82) detected marked changes in HSP27 phosphorylation at labor. Although total HSP27 levels were unchanged, HSP27-Ser15 was 3-fold higher at labor. Coimmunoprecipitation studies showed that HSP27 coprecipitates with alphaB crystallin and also smooth muscle alpha-actin. Coimmunofluorescence studies demonstrated a relocation of HSP27 from the perinuclear region to the actin cytoskeleton at labor. The functional significance of these changes was demonstrated in vitro where myometrial strips stimulated to contract with oxytocin exhibited increased HSP27-Ser15 phosphorylation. Our findings provide data consistent with a novel pathway regulating human myometrial contraction at labor and identify HSP27 and alphaB-crystallin as potential targets for future tocolytic design. PMID- 17916632 TI - Tyrosine phosphorylation of Munc18c regulates platelet-derived growth factor stimulated glucose transporter 4 translocation in 3T3L1 adipocytes. AB - Platelet-derived growth factor (PDGF) stimulation of skeletal muscle, cultured myotubes, and 3T3L1 adipocytes results in glucose transporter 4 (Glut4) translocation, albeit to a reduced level compared with insulin. To address the mechanism of PDGF action, we have determined that the Syntaxin 4 negative regulatory protein, Munc18c, undergoes PDGF-stimulated phosphorylation on tyrosine residue 521. The tyrosine phosphorylation of Munc18c on Y521 occurred concomitant with the dissociation of the Munc18c protein from Syntaxin 4 in a time frame consistent with Glut4 translocation. Moreover, expression of the wild type Munc18c protein did not inhibit PDGF-induced Glut4 translocation, whereas expression of Y521A-Munc18c mutant was inhibitory and failed to dissociate from Syntaxin 4. In contrast, expression of either wild-type Munc18c or the Y521A Munc18c mutant both resulted in a marked inhibition of insulin-stimulated Glut4 translocation. Together, these data demonstrate that one mechanism accounting for the PDGF induction of Glut4 translocation is the suppression of the Munc18c negative regulation of Syntaxin 4 function. PMID- 17916633 TI - Exogenous ghrelin attenuates the progression of chronic hypoxia-induced pulmonary hypertension in conscious rats. AB - Chronic exposure to hypoxia, a common adverse consequence of most pulmonary disorders, can lead to a sustained increase in pulmonary arterial pressure (PAP), right ventricular hypertrophy, and is, therefore, closely associated with heart failure and increased mortality. Ghrelin, originally identified as an endogenous GH secretagogue, has recently been shown to possess potent vasodilator properties, likely involving modulation of the vascular endothelium and its associated vasoactive peptides. In this study we hypothesized that ghrelin would impede the pathogenesis of pulmonary arterial hypertension during chronic hypoxia (CH). PAP was continuously measured using radiotelemetry, in conscious male Sprague Dawley rats, in normoxia and during 2-wk CH (10% O(2)). During this hypoxic period, rats received a daily sc injection of either saline or ghrelin (150 microg/kg). Subsequently, heart and lung samples were collected for morphological, histological, and molecular analyses. CH significantly elevated PAP in saline-treated rats, increased wall thickness of peripheral pulmonary arteries, and, consequently, induced right ventricular hypertrophy. In these rats, CH also led to the overexpression of endothelial nitric oxide synthase mRNA and protein, as well as endothelin-1 mRNA within the lung. Exogenous ghrelin administration attenuated the CH-induced overexpression of endothelial nitric oxide synthase mRNA and protein, as well as endothelin-1 mRNA. Consequently, ghrelin significantly attenuated the development of pulmonary arterial hypertension, pulmonary vascular remodeling, and right ventricular hypertrophy. These results demonstrate the therapeutic benefits of ghrelin for impeding the pathogenesis of pulmonary hypertension and right ventricular hypertrophy, particularly in subjects prone to CH (e.g. pulmonary disorders). PMID- 17916634 TI - Ethanol acutely stimulates islet blood flow, amplifies insulin secretion, and induces hypoglycemia via nitric oxide and vagally mediated mechanisms. AB - Hypoglycemia induced by alcohol ingestion is a well-known problem in diabetic patients. However, the mechanisms underlying this phenomenon have largely remained elusive. Because insulin secretion in vivo can be rapidly tuned by changes in pancreatic microcirculation, we evaluated the influence of acute alcohol administration on pancreatic islet blood flow (IBF), and dynamic changes in insulin secretion and glycemia in the rat. Ethanol (10%) or saline was iv injected as a bolus into Wistar rats, yielding serum ethanol concentrations of approximately 8 mmol/liter. Measurements of pancreatic blood flow (PBF) were performed by a microsphere technique in combination with a freeze-thawing technique after 10-min injection. Ethanol preferentially and significantly increased pancreatic IBF approximately 4-fold, whereas not influencing whole PBF. The alcohol also augmented late-phase insulin secretion and induced late hypoglycemia upon ip glucose tolerance tests. The nitric oxide synthase inhibitor N-w-nitro-L-arginine methyl ester and atropine prevented the increased pancreatic IBF, enhanced insulin secretion, and hypoglycemia evoked by ethanol. Thus, our findings demonstrate that ethanol acutely exerts substantial influences on pancreatic microcirculation by evoking a massive redistribution of PBF from the exocrine into the endocrine part via mechanisms mediated by nitric oxide and vagal stimuli, augmenting late-phase insulin secretion, and thereby evoking hypoglycemia. This effect may in part underlie the well-known hypoglycemic properties of alcohol in diabetic patients or in alcoholics with hepatic failure. PMID- 17916635 TI - Expression of Delta-like protein 4 in the human endometrium. AB - Activation of Delta-Notch signaling pathway promotes the development of the vascular system in embryo, normal adult tissues, and cancerous lesions. Delta and Notch genes are known to be expressed in endothelial cells, and little is known of their expression beyond the vascular system. The purpose of this study was to investigate whether Delta gene would be expressed in cells of the uterine endometrium. In this study, we found that the human endometrial cells expressed one of the Delta ligands, Delta-like 4 protein (Dll4). Dll4 was expressed in human endometrium in a spatiotemporal fashion. Immunohistochemistry studies showed the cytoplasm as well as membrane staining with apical localization both in the luminal and glandular epithelium and moderate diffuse staining in the cytoplasm of the stromal cells. Western blot analysis showed that the size of the endometrial Dll4 was identical to that in the human umbilical endothelial cells. The expression of Dll4 mRNA in human endometrial cells was quantitatively determined by real-time PCR. Dll4 mRNA expressed in the glandular epithelium showed large variations, and it was significantly elevated in the mid and late proliferative and early secretory endometrium. Endometrial stromal cells contained less Dll4 mRNA and had no clear correlation with the menstrual cycle. The effect of hormones was studied in the primary culture of isolated glandular epithelial and stromal cells. In glandular cells, estradiol had little effect, and medroxyprogesterone acetate significantly reduced the mRNAs compared with that of control. Relaxin induced the Dll4 mRNA. In stromal cells, both estradiol and medroxyprogesterone acetate reduced the Dll4 mRNA. To our knowledge, this is the first report of the expression of Dll4 in the endometrium. We propose that endometrial Dll4 may enhance the development of the endometrial microvascular system and facilitate the implantation of blastocyst in a fertile cycle. PMID- 17916636 TI - Comparative proteomic analysis of NaCl stress-responsive proteins in Arabidopsis roots. AB - NaCl stress is a major abiotic stress limiting the productivity and the geographical distribution of many plant species. Roots are the primary site of salinity perception. To understand better NaCl stress responses in Arabidopsis roots, a comparative proteomic analysis of roots that had been exposed to 150 mM NaCl for either 6 h or 48 h was conducted. Changes in the abundance of protein species within roots were examined using two-dimensional electrophoresis. Among the >1000 protein spots reproducibly detected on each gel, the abundance of 112 protein spots decreased and 103 increased, at one or both time points, in response to NaCl treatment. Through liquid-chromatography-tandem mass spectrometry, identity was assigned to 86 of the differentially abundant spots. The proteins identified included many previously characterized stress-responsive proteins and others related to processes including scavenging for reactive oxygen species; signal transduction; translation, cell wall biosynthesis, protein translation, processing and degradation; and metabolism of energy, amino acids, and hormones. At the resolution of individual genes and proteins, poor statistical correlation (6 h, r= -0.13; 48 h, r=0.11) of these protein expression data with previous microarray results was detected, supporting the concept that post-transcriptional regulation plays an important role in stress-responsive gene expression, and highlighting the need for combined transcriptomic and proteomic analyses. PMID- 17916637 TI - Gene expression profiles for cell wall-modifying proteins associated with soybean cyst nematode infection, petiole abscission, root tips, flowers, apical buds, and leaves. AB - Changes in transcript accumulation for cell wall-modifying proteins were examined in excised soybean root pieces colonized by soybean cyst nematodes (SCN), Heterodera glycines, using RT-PCR and soybean Affymetrix GeneChips. Sequence specific PCR primer pairs were prepared from sequence data for core sequences in the GenBank soybean database and consensus sequences derived from the assembly of soybean ESTs. In addition, to identify previously uncharacterized soybean transcripts, degenerate primers were prepared for conserved motifs in cellulases (endo-1,4-beta-glucanases, EGases) and polygalacturonases (PGs) and these were used to amplify segments of transcripts that were then extended with 3' and 5' RACE. Several novel EGase and PG transcripts were identified. Gene expression patterns were determined by real-time RT-PCR for 11 EGases, three expansins (EXPs), 14 PGs, two pectate lyases (PLs), and two xyloglucan endotransglucosylase/hydrolases (XTHs) in soybean roots inoculated with SCN, non inoculated roots, serial dissections of root tips, leaf abscission zones, flowers, apical buds, and expanding leaves. A large number of genes associated with cell wall modifications are strongly up-regulated in root pieces colonized by SCN. However, in contrast to most of the transcripts for cell wall proteins, two XTH transcripts were specifically down-regulated in the colonized root pieces. Gene expression in serial dissections of root tips (0-2 mm, and 2-7 mm) and whole roots indicate that the SCN up-regulated genes are associated with a wide range of developmental processes in roots. Also of interest, many of the cDNAs examined were up-regulated in petiole abscission zones induced to abscise with ethylene. PMID- 17916638 TI - Expression of ASCORBATE PEROXIDASE 8 in roots of rice (Oryza sativa L.) seedlings in response to NaCl. AB - Reactive oxygen species are thought to play an important role in NaCl stress. Therefore, the expression patterns of the gene family encoding the H(2)O(2) scavenging enzyme ascorbate peroxidase (APx; EC1.11.1.11) were analysed in roots of etiolated rice (Oryza sativa L.) seedlings in response to NaCl stress. Applying semi-quantitative RT-PCR, the mRNA levels were quantified for two cytosolic (OsAPx1 and OsAPx2), two peroxisomal (OsAPx3 and OsAPx4), and four chloroplastic (OsAPx5, OsAPx6, OsAPx7, and OsAPx8) isoforms identified in the rice genome. NaCl at 150 mM and 200 mM increased the expression of OsAPx8 and the activities of APx, but had no effect on the expression of OsAPx1, OsAPx2, OsAPx3, OsAPx4, OsAPx5, OsAPx6, and OsAPx7 in rice roots. However, NaCl at 300 mM up regulated OsAPx8 expression, increased APx activity, and down-regulated OsAPx7 expression, but had no effect on the expression of OsAPx1, OsAPx2, OsAPx3, OsAPx4, OsAPx5, and OsAPx6. The accumulation of abscisic acid (ABA) in response to NaCl was observed in rice roots. Exogenously applied ABA also specifically enhanced the expression of OsAPx8 in rice roots. The accumulation of ABA in rice roots in response to NaCl was inhibited by fluridone (Flu), an inhibitor of carotenoid biosynthesis. Flu treatment also suppressed NaCl-enhanced OsAPx8 expression and APx activity. The effect of Flu on the expression of OsAPx8 and increase in APx activity was reversed by the application of ABA. It appears that NaCl-enhanced expression of OsAPx8 in rice roots is mediated through an accumulation of ABA. Evidence is provided to show that Na(+) but not Cl(-) is required for enhancing OsAPx8 expression, APx activity, and ABA accumulation in rice roots treated with NaCl. H(2)O(2) treatment resulted in an enhancement of OsAPx8 induction but no accumulation of ABA. Diphenylene iodonium treatment, which is known to inhibit NaCl-induced accumulation of H(2)O(2) in rice roots, did not suppress OsAPx8 induction and ABA accumulation by NaCl. It appears that H(2)O(2) is not involved in the regulation of NaCl-induced OsAPx8 expression in rice roots. PMID- 17916639 TI - Decreased shoot stature and grain alpha-amylase activity following ectopic expression of a gibberellin 2-oxidase gene in transgenic wheat. AB - Ectopic expression of a gibberellin 2-oxidase gene (PcGA2ox1) decreased the content of bioactive gibberellins (GAs) in transgenic wheat, producing a range of dwarf plants with different degrees of severity. In at least one case, a single transformation event gave rise to T(1) plants with different degrees of dwarfism, the phenotypes being stably inherited over at least four generations. The dwarf phenotype, which included dark-green leaves, increased tillering and, in severe cases, a prostrate growth habit, was replicated by the application of a GA biosynthesis inhibitor to the wild type. Ear rachis length, grain set, and grain size were also decreased in the wheat transformants, compared with an azygous (null) line. The extent of post-germination alpha-amylase production in grains reflected the severity of the shoot phenotype of the transformants and both developmental processes were restored to normal by the application of gibberellic acid (GA(3)). Expression of two GA biosynthesis genes (TaGA20ox1 and TaGA3ox2) was up-regulated, and that of two alpha-amylase gene families (alpha-Amy1 and alpha-Amy2) down regulated, in scutella of semi-dwarf lines, compared with controls. The marked decline in transcript abundance of both alpha-amylase gene families in aleurone was associated with a decreased content of bioactive GAs in grains of the semi-dwarf lines. PMID- 17916640 TI - The role of mitochondrial and oxidative injury in BDE 47 toxicity to human fetal liver hematopoietic stem cells. AB - The polybrominated diphenyl ethers (PBDEs) are a group of flame retardants whose residues have markedly increased in the environment and in human tissues during the last decade. Of the various congeners, BDE 47 (2,2',4,4'-tetrabromodiphenyl ether) is typically the predominant congener observed in fish and wildlife samples, as well as in human tissues. Several studies indicate in utero transfer of PBDEs during pregnancy with residues accumulating in fetal tissues, and thus the potential for BDE 47-mediated injury in utero is of concern. In this study, we examined the mechanisms of BDE 47-mediated injury to primary human fetal liver hematopoietic stem cells (HSCs), which comprise a large proportion of fetal hepatic cells and play a key role in hematopoiesis during fetal development. Incubation of fetal liver HSCs with BDE 47 led to a loss of mitochondrial membrane potential and the onset of apoptosis. These effects were observed in the low micromolar range of BDE 47 exposures. At higher concentrations, BDE 47 elicited a loss of viability, which was accompanied by the generation of reactive oxygen species and peroxidation of HSC lipids. Preincubation of fetal liver HSCs with N-acetylcysteine, a glutathione (GSH) precursor, caused an increase in cellular GSH concentrations, restored mitochondrial redox status, and ameliorated the toxicity of BDE 47. BDE 47-mediated cytotoxicity or oxidative injury was not evident at the lower concentrations (< 1microM). Collectively, these data support a role for oxidative stress in the cytotoxicity of BDE 47 and indicate that oxidative stress-associated biomarkers may be useful in assessing the sublethal effects of BDE 47 toxicity in other models. However, the fact that BDE 47 undergoes a concentration-dependent accumulation in other primary cells in media that can underestimate cellular concentrations (W. R. Mundy et al., 2004, Toxicol. Sci. 82, 164-169) suggests that the HSC cell injury observed in our study may be of less relevance to human in utero PBDE exposures. PMID- 17916641 TI - Breed relationships facilitate fine-mapping studies: a 7.8-kb deletion cosegregates with Collie eye anomaly across multiple dog breeds. AB - The features of modern dog breeds that increase the ease of mapping common diseases, such as reduced heterogeneity and extensive linkage disequilibrium, may also increase the difficulty associated with fine mapping and identifying causative mutations. One way to address this problem is by combining data from multiple breeds segregating the same trait after initial linkage has been determined. The multibreed approach increases the number of potentially informative recombination events and reduces the size of the critical haplotype by taking advantage of shortened linkage disequilibrium distances found across breeds. In order to identify breeds that likely share a trait inherited from the same ancestral source, we have used cluster analysis to divide 132 breeds of dog into five primary breed groups. We then use the multibreed approach to fine-map Collie eye anomaly (cea), a complex disorder of ocular development that was initially mapped to a 3.9-cM region on canine chromosome 37. Combined genotypes from affected individuals from four breeds of a single breed group significantly narrowed the candidate gene region to a 103-kb interval spanning only four genes. Sequence analysis revealed that all affected dogs share a homozygous deletion of 7.8 kb in the NHEJ1 gene. This intronic deletion spans a highly conserved binding domain to which several developmentally important proteins bind. This work both establishes that the primary cea mutation arose as a single disease allele in a common ancestor of herding breeds as well as highlights the value of comparative population analysis for refining regions of linkage. PMID- 17916642 TI - Lateral gene transfer between obligate intracellular bacteria: evidence from the Rickettsia massiliae genome. AB - Rickettsia massiliae is a tick-borne obligate intracellular alpha-proteobacteria causing spotted fever in humans. Here, we present the sequence of its genome, comprising a 1.3-Mb circular chromosome and a 15.3-kb plasmid. The chromosome exhibits long-range colinearity with the other Spotted Fever Group Rickettsia genomes, except for a large fragment specific to R. massiliae that contains 14 tra genes presumably involved in pilus formation and conjugal DNA transfer. We demonstrate that the tra region was acquired recently by lateral gene transfer (LGT) from a species related to Rickettsia bellii. Further analysis of the genomic sequences identifies additional candidates of LGT between Rickettsia. Our study indicates that recent LGT between obligate intracellular Rickettsia is more common than previously thought. PMID- 17916643 TI - Biological role, protein expression, subcellular localization, and oxidative stress response of paraoxonase 2 in the intestine of humans and rats. AB - Oxidative stress is a cardinal manifestation of various intestinal disorders. However, very little knowledge is available on the intestine's inherent defense mechanisms against free radicals. This study was designed to determine the protein expression, subcellular localization and oxidative stress response of paraoxonase 2 (PON2), a member of a powerful antioxidant family in human and rat intestine. Biochemical and ultrastructural experiments all showed a substantial expression of PON2 in human and rat intestine. Western blot analysis disclosed higher levels of PON2 in the jejunum than in the duodenum, ileum, and colon. Cell fractionation revealed a predominant PON2 association with microsomes and lysosomes in the human jejunum, which differed from that in rats. PON2 was detected in the intestine as early as week 15 of gestation and was significantly increased by week 20. Iron ascorbate-mediated lipid peroxidation induced a marked decrease in PON2 expression in intestinal specimens coincidental to an abundant rise in malondialdehyde (MDA). On the other hand, preincubation with potent antioxidants, such as butylated hydroxytoluene, Trolox, and N-acetylcysteine, prevented iron-ascorbate-generating PON2 reduction in parallel with MDA suppression. Finally, the preincubation of permeabilized Caco-2 cells with purified PON2 led to a protection against iron-ascorbate-induced lipid peroxidation. These observations demonstrate that the human intestine is preferentially endowed with a marked PON2 expression compared with the rat intestine and this expression shows a developmental and intracellular pattern of distribution. Furthermore, our observations suggest PON2 protective effects against prooxidant stimuli in the small intestine. PMID- 17916644 TI - Early growth response-1 contributes to galactosamine/lipopolysaccharide-induced acute liver injury in mice. AB - Early growth response (Egr)-1 is a transcription factor that regulates genes involved in inflammation, innate and adaptive immunity, coagulation, and wound healing; however, little is known about the role of Egr-1 in acute liver injury. We tested the hypothesis that Egr-1 is involved in acute liver injury induced by galactosamine/lipopolysaccharide (GalN/LPS). GalN/LPS exposure biphasically increased hepatic egr-1 mRNA accumulation at 1 h and again at 4-5.5 h after treatment in wild-type mice. Within 4-5.5 h after GalN/LPS exposure, wild-type mice exhibited histological evidence of hepatocyte injury, cell death, and extensive areas of hemorrhage, as well as increased plasma alanine aminotransferase activities. In contrast, these parameters were largely attenuated in egr-1(-/-) mice. The initial expression of tumor necrosis factor alpha, macrophage inflammatory protein-2, monocyte chemoattractant protein-1, and intercellular adhesion molecule-1 mRNA or protein was equivalent between genotypes at 1 h after GalN/LPS administration. However, at subsequent time points, hepatic expression of these genes was decreased in egr-1(-/-) compared with wild-type mice. In addition, neutrophil extravasation from hepatic sinusoids into the liver parenchyma was decreased in egr-1(-/-) compared with wild-type mice 4 h after GalN/LPS. Whereas caspase-3 activation and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive nuclei were detected in wild-type mice at 4 and 5.5 h after GalN/LPS administration, respectively, these markers of apoptosis were delayed in egr-1(-/-) mice. Delayed development of apoptosis was associated with an extension of survival by 1 h in egr-1(-/-) compared with wild-type mice. These data demonstrate that Egr-1 plays an important role in acceleration of hepatic inflammation, apoptosis, and subsequent mortality in GalN/LPS-induced acute liver injury. PMID- 17916646 TI - Duodenal brush border intestinal alkaline phosphatase activity affects bicarbonate secretion in rats. AB - We hypothesized that duodenal HCO(3)(-) secretion alkalinizes the microclimate surrounding intestinal alkaline phosphatase (IAP), increasing its activity. We measured AP activity in rat duodenum in situ in frozen sections with the fluorogenic substrate ELF-97 phosphate and measured duodenal HCO(3)(-) secretion with a pH-stat in perfused duodenal loops. We examined the effects of the IAP inhibitors L-cysteine or L-phenylalanine (0.1-10 mM) or the tissue nonspecific AP inhibitor levamisole (0.1-10 mM) on AP activity in vitro and on acid-induced duodenal HCO(3)(-) secretion in vivo. AP activity was the highest in the duodenal brush border, decreasing longitudinally to the large intestine with no activity in stomach. Villous surface AP activity measured in vivo was enhanced by PGE(2) intravenously and inhibited by luminal L-cysteine. Furthermore, incubation with a pH 2.2 solution reduced AP activity in vivo, whereas pretreatment with the cystic fibrosis transmembrane regulator (CFTR) inhibitor CFTR(inh)-172 abolished AP activity at pH 2.2. L-Cysteine and L-phenylalanine enhanced acid-augmented duodenal HCO(3)(-) secretion. The nonselective P2 receptor antagonist suramin (1 mM) reduced acid-induced HCO(3)(-) secretion. Moreover, L-cysteine or the competitive AP inhibitor glycerol phosphate (10 mM) increased HCO(3)(-) secretion, inhibited by suramin. In conclusion, enhancement of the duodenal HCO(3)(-) secretory rate increased AP activity, whereas inhibition of AP activity increased the HCO(3)(-) secretory rate. These data support our hypothesis that HCO(3)(-) secretion increases AP activity by increasing local pH at its catalytic site and that AP hydrolyzes endogenous luminal phosphates, presumably ATP, which increases HCO(3)(-) secretion via activation of P2 receptors. PMID- 17916645 TI - CD4 T cells activated in the mesenteric lymph node mediate gastrointestinal food allergy in mice. AB - A localized Th2 milieu has been observed in the intestine of subjects with food allergic disorders; however, the role of T cells in the pathophysiology of these disorders remains poorly understood. Our aim was to examine sites of T cell activation in response to food challenge, identify potential factors responsible for T cell recruitment to the gut, and determine the role of T cells in disease. BALB/c mice were systemically sensitized to ovalbumin (OVA) and repeatedly fed with OVA to induce allergic diarrhea. Local cytokine and chemokine expressions were assessed by quantitative PCR, and cytokine secretion levels in the mesenteric lymph node (MLN) were determined by ELISA. Homing molecule expression was determined by flow cytometry, and the role of CD4(+) T cells in promoting disease was tested by adoptive transfer. Mice developed diarrhea associated with changes in epithelial ion transport, mast cell infiltration, intestinal IgE secretion, and local upregulation of Th2 cytokines and the Th2 chemokines CCL1, CCL17, and CCL22 in the small intestine. T cell activation occurred in the MLN before symptom onset, and a single feed of OVA induced T cell proliferation, alpha(4)beta(7) upregulation, and CD62L downregulation. Cells from the MLN, including purified CD4(+) T cells, were able to transfer allergic diarrhea to naive mice. A gut-homing phenotype induced in the MLN and selective upregulation of Th2 chemoattractants are likely important factors in the gastrointestinal recruitment of pathological Th2-skewed CD4(+) T cells in food allergy. PMID- 17916647 TI - Effect of retinoic acid on cell proliferation and differentiation as well as on lipid synthesis, lipoprotein secretion, and apolipoprotein biogenesis. AB - Dietary vitamin A and its active metabolites are essential nutrients for many functions as well as potent regulators of gene transcription and growth. Although the epithelium of the small intestine is characterized by rapid and constant renewal and enterocytes play a central role in the absorption and metabolism of alimentary retinol, very little is known about the function of retinoids in the human gastrointestinal epithelium and mechanisms by which programs engage the cell cycle are poorly understood. We have assessed the effects of 10 microM 9- and 13-cis-retinoic acid (RA) on proliferation and differentiation processes, lipid esterification, apolipoprotein (apo) biogenesis and lipoprotein secretion along with nuclear factor gene transcription. Treatment of Caco-2 cells with RA at different concentrations and incubation periods revealed the reduction of thymidine incorporation in 60% preconfluent or 100% confluent cells. Concomitantly, RA 1) modulated D-type cyclins by reducing the mitogen-sensitive cyclin D1 and upregulating cyclin D3 expressions and 2) caused a trend of increase in p38 MAPK, which triggers CDX2, a central protein in cell differentiation. RA remained without effect on lipoprotein output and apo synthesis, even for apo A-I that possesses RARE in its promoter. RA, in combination with 22-hydroxycholesterol, could induce apo A-I gene expression without any impact on apo A-I mass. Only the gene expression of peroxisome proliferator-activated receptor (PPAR)beta, retinoic receptor (RAR)beta, and RARgamma was augmented and no alteration was noted in PPARalpha, PPARgamma, liver X receptor (LXR)alpha, LXRbeta, and retinoid X receptors. Taken together, these data highlight RA-induced cell differentiation via specific signaling without a significant impact on apo A-I synthesis. PMID- 17916649 TI - Fenofibrate inhibits intestinal Cl- secretion by blocking basolateral KCNQ1 K+ channels. AB - Fibrates are peroxisome proliferator-activated receptor-alpha (PPARalpha) ligands in widespread clinical use to lower plasma triglyceride levels. We investigated the effect of fenofibrate and clofibrate on ion transport in mouse intestine and in human T84 colonic adenocarcinoma cells through the use of short-circuit current (I(sc)) and ion flux analysis. In mice, oral administration of fenofibrate produced a persistent inhibition of cAMP-stimulated electrogenic Cl( ) secretion by isolated jejunum and colon without affecting electroneutral fluxes of (22)Na(+) or (86)Rb(+) (K(+)) across unstimulated colonic mucosa. When applied acutely to isolated mouse intestinal mucosa, 100 microM fenofibrate inhibited cAMP-stimulated I(sc) within 5 min. In T84 cells, fenofibrate rapidly inhibited approximately 80% the Cl(-) secretory responses to forskolin (cAMP) and to heat stable enterotoxin STa (cGMP) without affecting the response to carbachol (Ca(2+)). Both fenofibrate and clofibrate inhibited cAMP-stimulated I(sc) with an IC(50) approximately 1 muM, whereas other PPARalpha activators (gemfibrozil and Wy-14,643) were without effect. Membrane permeabilization experiments on T84 cells indicated that fenofibrate inhibits basolateral cAMP-stimulated K(+) channels (putatively KCNQ1/KCNE3) without affecting Ca(2+)-stimulated K(+) channel activity, whereas clofibrate inhibits both K(+) pathways. Fenofibrate had no effect on apical cAMP-stimulated Cl(-) channel activity. Patch-clamp analysis of HEK-293T cells confirmed that 100 microM fenofibrate rapidly inhibits K(+) currents associated with ectopic expression of human KCNQ1 with or without the KCNE3 beta-subunit. We conclude that fenofibrate inhibits intestinal cAMP stimulated Cl(-) secretion through a nongenomic mechanism that involves a selective inhibition of basolateral KCNQ1/KCNE3 channel complexes. Our findings raise the prospect of fenofibrate as a safe and effective antidiarrheal agent. PMID- 17916648 TI - Molecular mechanisms contributing to glutamine-mediated intestinal cell survival. AB - Glutamine, the most abundant amino acid in the bloodstream, is the preferred fuel source for enterocytes and plays a vital role in the maintenance of mucosal growth. The molecular mechanisms regulating the effects of glutamine on intestinal cell growth and survival are poorly understood. Here, we show that addition of glutamine (1 mmol/l) enhanced rat intestinal epithelial (RIE)-1 cell growth; conversely, glutamine deprivation increased apoptosis as noted by increased DNA fragmentation and caspase-3 activity. To delineate signaling pathways involved in the effects of glutamine on intestinal cells, we assessed activation of extracellular signal-related kinase (ERK), protein kinase D (PKD), and phosphatidylinositol 3-kinase (PI3K)/Akt, which are important pathways in cell growth and survival. Addition of glutamine activated ERK and PKD in RIE-1 cells after a period of glutamine starvation; inhibition of ERK, but not PKD, increased cell apoptosis. Conversely, glutamine starvation alone increased phosphorylated Akt; inhibition of Akt enhanced RIE-1 cell DNA fragmentation. The role of ERK was further delineated using RIE-1 cells stably transfected with an inducible Ras. Apoptosis was significantly increased following ERK inhibition, despite Ras activation. Taken together, these results identify a critical role for the ERK signaling pathways in glutamine-mediated intestinal homeostasis. Furthermore, activation of PI3K/Akt during periods of glutamine deprivation likely occurs as a protective mechanism to limit apoptosis associated with cellular stress. Importantly, our findings provide novel mechanistic insights into the antiapoptotic effects of glutamine in the intestine. PMID- 17916650 TI - Excitatory effects of synchronized intestinal electrical stimulation on small intestinal motility in dogs. AB - The aim of this study was to investigate effects of synchronized intestinal electrical stimulation (SIES) on small intestinal motility in dogs. Seventeen dogs were equipped with a duodenal cannula for the measurement of small bowel motility using manometry; an additional cannula was equipped in six of the dogs with 1.5 m distal to the first one for the measurement of small intestinal transit. Two pairs of bipolar electrodes were implanted on the small intestinal serosa with an interval of 5 cm; glucagon was used to induce postprandial intestinal hypomotility. Eleven dogs were used for the assessment of the small intestinal contractions in both fasting and fed states. The other six dogs were used for the measurement of small intestinal transit. We found that 1) SIES induced small intestinal contractions during phase I of the migrating motor complex (MMC) (contractile index or CI: 5.2 +/- 0.6 vs. 10.3 +/- 0.7, P = 0.003); 2) in the fed state, SIES significantly improved glucagon-induced small intestinal postprandial hypomotility (CI: 3.4 +/- 0.5 vs. 6.0 +/- 0.3, P = 0.03); 3) SIES significantly accelerated small intestinal transit delayed by glucagon (70.4 +/- 3.1 vs. 44.5 +/- 3.1 min, P < 0.01); 4) there was a negative correlation between the CI and transit time (r = -0.427, P = 0.048); and 5) the excitatory effect of SIES was blocked by atropine. SIES may have a therapeutic potential for treating patients with small intestinal disorders. PMID- 17916651 TI - Obstructive cholestasis induces TNF-alpha- and IL-1 -mediated periportal downregulation of Bsep and zonal regulation of Ntcp, Oatp1a4, and Oatp1b2. AB - Inverse acinar regulation of Mrp2 and 3 represents an adaptive response to hepatocellular cholestatic injury. We studied whether obstructive cholestasis (bile duct ligation) and LPS treatment affect the zonal expression of Bsep (Abcb11), Mrp4 (Abcc4), Ntcp (Slc10a1), and Oatp isoforms (Slco1a1, Slco1a4, and slco1b2) in rat liver, as analyzed by semiquantitative immunofluorescence. Contribution of TNF-alpha and IL-1beta to transporter zonation in obstructive cholestasis was studied by cytokine inactivation. In normal liver Bsep, Mrp4, Ntcp, and Oatp1a1 were homogeneously distributed in the acinus, whereas Oatp1a4 and Oatp1b2 expression increased from zone 1 to 3. Glutamine synthetase-positive pericentral hepatocytes exhibited markedly lower Oatp1a4 expression than the remaining zone 3 hepatocytes. In cholestatic liver Bsep and Ntcp immunofluorescence in periportal hepatocytes significantly decreased to 66 +/- 4% (P < 0.01) and 67 +/- 7% (P < 0.05), whereas it was not altered in pericentral hepatocytes. Oatp1a4 was significantly induced in hepatocytes with a primarily low expression, i.e., in periportal hepatocytes and in glutamine synthetase positive pericentral hepatocytes. Likewise, Oatp1b2 was upregulated in periportal hepatocytes. Mrp4 zonal induction was homogeneous. Inactivation of TNF-alpha and IL-1beta prevented periportal downregulation of Bsep. Recruitment of neutrophils and polymorphonuclear cells mainly occurred in the periportal zone. Likewise, IL 1beta induction was largely found periportally. No significant transporter zonation was seen following LPS treatment. In conclusion, zonal downregulation of Bsep in obstructive cholestasis is associated with portal inflammation and is mediated by TNF-alpha and IL-1beta. Periportal downregulation of Ntcp and induction of Oatp1a4 and Oatp1b2 may represent adaptive mechanisms to reduce cholestatic injury in hepatocytes with profound downregulation of Bsep and Mrp2. PMID- 17916652 TI - Prostaglandin E2 modulates TNF-alpha-induced MCP-1 synthesis in pancreatic acinar cells in a PKA-dependent manner. AB - Cyclooxygenase (COX)-2 is increased in human chronic pancreatitis. We recently demonstrated in a model of chronic pancreatitis (WBN/Kob rat) that inhibition of COX-2 activity reduces and delays pancreatic inflammation and fibrosis. Monocyte chemoattractant protein (MCP)-1 mRNA and PGE(2) were significantly reduced, correlating with a decreased infiltration of macrophages. MCP-1 plays an important role in the recruitment of macrophages to the site of tissue injury. The aim of our study is to identify mechanisms by which macrophages and acinar cells maintain an inflammatory reaction. The expression profile of E prostanoid receptors EP(1-4) and MCP-1 was analyzed by RT-PCR from pancreatic specimens and AR42J cells. MCP-1 secretion was detected by ELISA from rat pancreatic lobuli. We determined EP(1-4) mRNA levels in WBN/Kob rats with chronic pancreatic inflammation. Individual isoforms were highly increased in rat pancreas, concurrent with MCP-1 mRNA expression. In supernatants of pancreatic lobuli and AR42J cells, MCP-1 was detectable by ELISA. In the presence of TNF-alpha, MCP-1 was upregulated. Coincubation with PGE(2) enhanced the TNF-alpha-induced MCP-1 synthesis significantly. Similarly, TNF-alpha mRNA was synergistically upregulated by TNF-alpha and PGE(2). Furthermore, the synergistic effect of TNF alpha and PGE(2) was abolished by inhibition of PKA but not of PKC. We conclude that EP receptors are upregulated during chronic pancreatic inflammation. PGE(2) modulates the TNF-alpha-induced MCP-1 synthesis and secretion from acinar cells. This synergistic effect is controlled by PKA. This mechanism might explain the COX-2-dependent propagation of pancreatic inflammation. PMID- 17916653 TI - Prostaglandin F2alpha stimulates the expression and secretion of transforming growth factor B1 via induction of the early growth response 1 gene (EGR1) in the bovine corpus luteum. AB - In most mammals, prostaglandin F2alpha (PGF2alpha) is believed to be a trigger that induces the regression of the corpus luteum (CL), whereby progesterone synthesis is inhibited, the luteal structure involutes, and the reproductive cycle resumes. Studies have shown that the early growth response 1 (EGR1) protein can induce the expression of proapoptotic proteins, suggesting that EGR1 may play a role in luteal regression. Our hypothesis is that EGR1 mediates the actions of PGF2alpha by inducing the expression of TGF beta1 (TGFB1), a key tissue remodeling protein. The levels of EGR1 mRNA and protein were up-regulated in the bovine CL during PGF2alpha-induced luteolysis in vivo and in PGF2alpha-treated luteal cells in vitro. Using chemical and genetic approaches, the RAF/MAPK kinase (MEK) 1/ERK pathway was identified as a proximal signaling event required for the induction of EGR1 in PGF2alpha-treated cells. Treatment with PGF2alpha increased the expression of TGFB1 mRNA and protein as well as the binding of EGR1 protein to TGFB1 promoter in bovine luteal cells. The effect of PGF2alpha on TGFB1 expression was mimicked by a protein kinase C (PKC)/RAF/MEK1/ERK activator or adenoviral-mediated expression of EGR1. The stimulatory effect of PGF2alpha on TGFB1 mRNA and TGFB1 protein secretion was inhibited by blockade of MEK1/ERK signaling and by adenoviral-mediated expression of NAB2, an EGR1 binding protein that inhibits EGR1 transcriptional activity. Treatment of luteal cells with TGFB1 reduced progesterone secretion, implicating TGFB1 in luteal regression. These studies demonstrate that PGF2alpha stimulates the expression of EGR1 and TGFB1 in the CL. We suggest that EGR1 plays a role in the expression of genes whose cognate proteins coordinate luteal regression. PMID- 17916654 TI - Fibroblast growth factor receptor-1 signaling in pancreatic islet beta-cells is modulated by the extracellular matrix. AB - Maintenance of pancreatic beta-cell mass depends on extracellular stimuli that promote survival and proliferation. In the islet, these stimuli come from the beta-cell microenvironment and include extracellular matrix deposited by associated vascular endothelial cells. Fibroblast growth factor receptor-1 (FGFR1) has recently been implicated as a signaling pathway that is important for normal beta-cell function. We would like to understand how extracellular matrix and FGFR1 signaling interact to promote beta-cell survival and proliferation. To examine beta-cell-specific receptor responses, we created lentiviral vectors with rat insulin promoter-driven expression of Venus fluorescent protein-tagged full length (R1betav) and kinase-deficient (KDR1betav) FGFR1. Significant FGF-1 dependent activation of ERK1/2 was observed in betaTC3 cells, dispersed beta cells, and beta-cells in intact islets. This response was enhanced by R1betav expression and reduced by KDR1betav expression. Plating-dispersed beta-cells on collagen type IV resulted in enhanced expression of endogenous FGFR1 that was associated with sustained activation of ERK1/2. Conversely, plating cells on laminin reduced expression of FGFR1, and this reduction was associated with transient activation of ERK1/2. Addition of neutralizing antibodies to inhibit beta-cell attachment to laminin via alpha(6)-integrin increased high-affinity FGF 1-binding at the plasma membrane and resulted in sustained ERK1/2 activity similar to cells plated on collagen type IV. These data show that the FGF stimulated beta-cell response is negatively affected by alpha(6)-integrin binding to laminin and suggest regulation associated with vascular endothelial cell remodeling. PMID- 17916655 TI - Defective protein folding and intracellular retention of thyroglobulin-R19K mutant as a cause of human congenital goiter. AB - It has been suggested that a thyroglobulin (Tg)-R19K missense mutation may be a newly identified cause of human congenital goiter, which is surprising for this seemingly conservative substitution. Here, we have examined the intracellular fate of recombinant mutant Tg expressed in COS-7 cells. Incorporation of the R19K mutation largely blocked Tg secretion, and this mutant was approximately 90% degraded intracellularly over a 24-h period after synthesis. Before its degradation, the Tg-R19K mutant exhibited abnormally increased association with molecular chaperones BiP, calnexin, and protein disulfide isomerase, and was unable to undergo anterograde advance from the endoplasmic reticulum (ER) through the Golgi complex. Inhibitors of proteasomal proteolysis and ER mannosidase-I both prevented ER-associated degradation of the Tg-R19K mutant and increased its association with ER molecular chaperones. ER quality control around Tg residue 19 is not dependent upon charge but upon side-chain packing, because Tg-R19Q was efficiently secreted. Whereas a Tg mutant truncated after residue 174 folds sufficiently well to escape ER quality control, introduction of the R19K point mutation blocked its secretion. The data indicate that the R19K mutation induces local misfolding in the amino-terminal domain of Tg that has global effects on Tg transport and thyroid hormonogenesis. PMID- 17916656 TI - Neural correlates of proprioceptive integration in the contralesional hemisphere of very impaired patients shortly after a subcortical stroke: an FMRI study. AB - BACKGROUND: The effects of physiotherapy are difficult to assess in very impaired early stroke patients. OBJECTIVE: The aim of the study was to characterize the impact of 4 weeks of passive proprioceptive training of the wrist on brain sensorimotor activation after stroke. METHODS: Patients with a subcortical ischemic lesion of the pyramidal tract were randomly assigned to a control or a wrist-training group. All patients had a single pure motor hemiplegia with severe motor deficit. The control group (6 patients) underwent standard Bobath rehabilitation. The second, "trained," group (7 patients) received Bobath rehabilitation plus 4 weeks of proprioceptive training with daily passive calibrated wrist extension. Before and after the training period, patients were examined with validated clinical scales and functional MRI (fMRI) while executing a passive movement versus rest. The effect of standard rehabilitation on sensorimotor activation was assessed in the control group on the wrist, and the effect of standard rehabilitation plus proprioceptive training was assessed in the trained group. The effect of 4-week proprioceptive training alone was statistically evaluated by difference between groups. RESULTS: Standard rehabilitation along with natural recovery mainly led to increases in ipsilesional activation and decreases in contralesional activation. On the contrary, standard rehabilitation and paretic wrist proprioceptive training increased contralesional activation. Proprioceptive training produced change in the supplementary motor area (SMA), prefrontal cortex, and a contralesional network including inferior parietal cortex (lower part of BA 40), secondary sensory cortex, and ventral premotor cortex (PMv). CONCLUSION: We have demonstrated that purely passive proprioceptive training applied for 4 weeks is able to modify brain sensorimotor activity after a stroke. This training revealed fMRI change in the ventral premotor and parietal cortices of the contralesional hemisphere, which are secondary sensorimotor areas. Recent studies have demonstrated the crucial role of these areas in severely impaired patients. We propose that increased contralesional activity in secondary sensorimotor areas likely facilitates control of recovered motor function by simple proprioceptive integration in those patients with poor recovery. PMID- 17916657 TI - Short-term changes in and predictors of participation of older adults after stroke following acute care or rehabilitation. AB - BACKGROUND: Stroke can lead to restrictions in participation in daily activities and social roles. Although considered an important rehabilitation outcome, little is known about participation after stroke and its predictors, and about the differences associated with the types of services provided following stroke. OBJECTIVE: The aims of this study were 1) to follow and compare changes in participation of older adults discharged home after stroke from acute care or postacute rehabilitation, and 2) to identify the best predictors of participation after stroke from physical, cognitive, perceptual, and psychological ability measures taken shortly after discharge. METHODS: Level of participation in daily activities and social roles of 197 older adults who had a stroke was evaluated at 2 to 3 weeks (T1), 3 months (T2), and 6 months (T3) after being discharged home from acute care (n = 86) or rehabilitation (n = 111). Physical, cognitive, perceptual, and psychological abilities were assessed at T1. RESULTS: A significant increase in participation was found over time for both groups, mainly in the first 3 months. The best predictors of participation differed between the groups and between the daily activities and social roles domains. Walking and acceptance of the stroke or fewer depressive symptoms were the best predictors of the level of participation after stroke. CONCLUSIONS: Participation was not optimal at discharge because it continued to increase after the return home. The importance of psychological factors in participation after stroke is undeniable. Many predictors are amenable to interventions. PMID- 17916658 TI - The influence of hand dominance on the response to a constraint-induced therapy program following stroke. AB - BACKGROUND: Following stroke it is common to exhibit deficits in mobility of the upper extremity. Constraint-induced therapy (CIT) is a rehabilitation technique used to promote use of the more affected hand via constraint of the less affected hand. One factor that could impact the outcome following CIT is hand dominance. Years of preferred use of one hand may give individuals with the dominant hand affected by stroke an advantage in improving the mobility of the more affected hand compared to those individuals with the nondominant hand affected by stroke. In addition, the diminished use of the less affected hand during CIT may also create changes. OBJECTIVE: Our goal was to better understand how hand dominance may influence the response to a CIT program both cortically and behaviorally in both the more affected hand and less affected constrained hand. METHODS: A repeated measures design with a double baseline was used to assess changes in clinical tests and functional magnetic resonance imaging (fMRI) in individuals with their dominant or nondominant hand affected by stroke involved in a CIT program. RESULTS: No significant differences were found between groups in their responses to CIT. Overall subjects demonstrated behavioral and cortical changes with the more affected hand and the less affected constrained hand did not significantly change. CONCLUSION: CIT promotes improvement of the more affected hand particularly on complex tests without decrements to the less affected constrained hand. Cortically, statistically significant changes in activation were noted after the intervention for the more affected hand; no changes were noted with the less affected constrained hand. PMID- 17916660 TI - Workload at the heart rate of 100 beats/min and mortality in middle-aged men with known or suspected coronary heart disease. AB - OBJECTIVE: To investigate whether a workload which an individual is able to perform at the heart rate (HR) of 100 beats/min (WL(100)) independently predicts mortality in middle-aged men with known or suspected coronary heart disease (CHD). DESIGN: Prospective population-based study based on 365 middle-aged men with known or suspected CHD at baseline. RESULTS: During an average follow-up of 11.1 years, there were 75 deaths (20.5%). In Cox multivariable models mortality increased by 72% (95% CI 32% to 122%, p<0.001) with 1 SD (34 Watts) decrement in WL(100) after adjustment for age, examination year, alcohol consumption, body mass index, cigarette smoking, cardiac insufficiency, history of myocardial infarction, diabetes, myocardial ischaemia during exercise test, serum low density lipoprotein and high-density lipoprotein cholesterol, systolic and diastolic blood pressure at rest, testing protocol, and use of HR-lowering medication. The risk of death was 2.4 (95% CI 1.5 to 4.0, p<0.001) times higher in 130 men with WL(100) <55 W than in 235 men with WL(100) >or=55 W. In men using and not using HR-lowering medication the risk of death increased 72% (95% CI 14% to 163%, p = 0.01), and 54% (95% CI 14% to 108%, p = 0.005) with 1 SD decrement in WL(100), respectively. WL(100 )improved the predictive power of the adjusted Cox models including other HR and exercise test variables. CONCLUSIONS: WL(100) predicts mortality in men with known or suspected CHD. The association of WL(100) with mortality was not explained by other well-established HR and exercise test variables. WL(100) is derived from a submaximal test which avoids the cardiovascular risks associated with a high-intensity exertion. PMID- 17916661 TI - Performance of the QRISK cardiovascular risk prediction algorithm in an independent UK sample of patients from general practice: a validation study. AB - AIM: To assess the performance of the QRISK score for predicting cardiovascular disease (CVD) in an independent UK sample from general practice and compare with the Framingham score. DESIGN: Prospective open cohort study. SETTING: UK general practices contributing to the THIN and QRESEARCH databases. COHORT: The THIN validation cohort consisted of 1.07 million patients, aged 35-74 years registered at 288 THIN practices between 1 January 1995 and 1 April 2006. The QRESEARCH validation cohort consisted of 0.61 million patients from 160 practices (one third of the full database) with data until 1 January 2007. Patients receiving statins, those with diabetes or CVD at baseline were excluded. END POINT: First diagnosis of CVD (myocardial infarction, coronary heart disease (CHD), stroke and transient ischaemic attack) recorded on the clinical computer system during the study period. EXPOSURES: Age, sex, smoking status, systolic blood pressure, total/high-density lipoprotein cholesterol ratio, body mass index, family history of premature CHD, deprivation and antihypertensive medication. RESULTS: Characteristics of both cohorts were similar, except that THIN patients were from slightly more affluent areas and had lower recording of family history of CHD. QRISK performed better than Framingham for every discrimination and calibration statistic in both cohorts. Framingham overpredicted risk by 23% in the THIN cohort, while QRISK underpredicted risk by 12%. CONCLUSION: This analysis demonstrated that QRISK is better calibrated to the UK population than Framingham and has better discrimination. The results suggest that QRISK is likely to provide more appropriate risk estimates than Framingham to help identify patients at high risk of CVD in the UK. PMID- 17916662 TI - Chronic treatment with tetrahydrobiopterin reverses endothelial dysfunction and oxidative stress in hypercholesterolaemia. AB - BACKGROUND: Reduced availability of tetrahydrobiopterin (BH(4)), an essential cofactor of nitric oxide (NO) synthase (NOS), decreases NO production and increases reactive oxygen species. Both mechanisms contribute to atherosclerotic vascular disease. Although acute supplementation of BH(4) improves endothelial dysfunction, the effect of chronic BH(4) in humans is unknown. OBJECTIVE: To investigate the effect of chronic BH(4) supplementation on endothelial function and oxidative stress in hypercholesterolaemia. DESIGN: Randomised double-blind, placebo-controlled trial. SETTING: University Hospital. PATIENTS: 22 hypercholesterolaemic patients (low-density lipoprotein (LDL) >4.5 mmol/l) were randomised to 4 weeks of oral BH(4) (400 mg twice daily) or placebo. Age-matched healthy volunteers served as controls. MAIN OUTCOME MEASURES: Endothelium dependent and -independent vasodilatation was assessed by venous occlusion plethysmography. To elucidate the mechanisms of BH(4) effect, NO release and superoxide anion (O(2)(-)) production were measured in human aortic endothelial cells exposed to native LDL (2.6 mmol cholesterol/l). RESULTS: BH(4) plasma levels were significantly increased by oral supplementation. NO-mediated vasodilatation to acetylcholine was reduced in patients compared with controls and restored by BH(4). No effect of BH(4) on endothelium-independent vasodilatation was seen. Furthermore, 8-F(2 )isoprostane plasma levels, a marker of vascular oxidative stress, were reduced by BH(4). In LDL-treated endothelial cells, BH(4) levels and NO release were reduced and O(2)(-) production increased compared with control cells. Exogenous BH(4) normalised NO and O(2)(-) production. CONCLUSIONS: In hypercholesterolaemia, endothelial dysfunction and oxidative stress can be reversed by chronic oral treatment with BH(4). Thus, BH(4) availability is essential for maintaining NO synthesis and low O(2)(-) production by endothelial NOS in vivo, and may provide a rational therapeutic approach to prevent cardiovascular disease. PMID- 17916663 TI - Long-term outcome of low-risk patients attending a rapid-assessment chest pain clinic. AB - OBJECTIVE: To examine the long-term outcome of patients evaluated in a rapid assessment chest pain clinic (RACPC): are "low-risk" patients safely reassured? DESIGN: Retrospective cohort study. SETTING: Staff grade-led RACPC in an urban teaching hospital. PARTICIPANTS: 3378 patients (51% male), attending the RACPC between April 1996 and February 2000. MAIN OUTCOME MEASURES: Death, coronary mortality, morbidity and revascularisation over a median follow-up of 6 years. Coronary standardised mortality ratio (SMR). RESULTS: 2036 (60.3%) patients were categorised as "low risk", 957 (28.3%) as having "stable coronary artery disease" and 214 (6.3%) as being an "acute coronary syndrome". During the study, 3.6% of patients in the low risk category, 11.9% in the stable coronary artery disease category and 24.6% in the acute coronary syndrome category died from coronary artery disease or had a myocardial infarction. 5.5%, 18.2% and 18.4%, respectively, died from any cause. Compared to the local population (coronary SMR = 100), our "low risk/non-coronary chest pain" cohort had a coronary SMR of 51 (95% CI 31 to 83), the "stable coronary artery disease" cohort 240 (187 to 308) and the "acute coronary syndrome" cohort 780 (509 to 1196). CONCLUSION: The RACPC was effective at triaging patients with chest pain. Patients identified as at "low risk" were unlikely to have an adverse coronary outcome and were appropriately reassured. PMID- 17916664 TI - Smoking status, sports participation and mortality from coronary heart disease. AB - BACKGROUND: Since smoking and exercise have opposite effects on coronary risk factors, the hypothesis was proposed that smoking might weaken the protective effect of exercise on prevention of coronary heart disease. OBJECTIVE: To determine the effect of smoking on the relationship between sports participation and mortality from coronary heart disease. DESIGN: Population-based prospective cohort study in Japan. PARTICIPANTS: A total of 76 832 Japanese men and women, aged 40-79 years with no history of stroke, coronary heart disease, or cancer, completed a self-administered questionnaire between 1988 and 1990. MAIN OUTCOME MEASURES: Systematic mortality surveillance was carried out through 2003, and 638 deaths from coronary heart disease (496 myocardial infarction) were identified. RESULTS: People who reported the longest time in sports participation (>or=5 hours/week) had an approximately 50-80% lower age-adjusted risk of mortality from coronary heart disease compared with those in the second lowest category (1-2 hours/week) among never and ex-smokers, but no association was found among current smokers. Adjustment for known risk factors and exclusion of subjects who died within 2 years of the baseline inquiry did not substantially alter these associations. The multivariable hazard ratios (95% confidence interval) of coronary heart disease for the >or=5 hours/week versus 1-2 hours/week of sports participation were 0.44 (0.23 to 0.86) among never smokers, 0.18 (0.05 to 0.60) among ex-smokers, and 0.82 (0.47 to 1.40) among current smokers. Similar associations were found for men and women. CONCLUSIONS: Smoking may reduce the beneficial effect of sports participation for reduction of fatal coronary heart disease. PMID- 17916666 TI - Quantitative dobutamine stress echocardiography for the early detection of cardiac allograft vasculopathy in heart transplant recipients. AB - BACKGROUND: A non-invasive method to detect the presence of cardiac allograft vasculopathy (CAV) remains an important goal in clinical cardiology. OBJECTIVE: To assess the value of quantitative dobutamine stress echocardiography (DSE) for the early detection of CAV. METHODS: 42 heart transplant recipients underwent DSE with acquisition of both conventional two-dimensional and colour tissue Doppler data. All studies were analysed conventionally and quantitatively using regional deformation parameters-that is, peak systolic longitudinal strain (in(peak sys)), strain rate (SR(peak sys)) and post-systolic strain index. Myocardial segments were classified as normal, mildly abnormal or severely abnormal based on correlative angiographic findings. RESULTS: At baseline, in(peak sys) was significantly lower in severely abnormal segments than in normal ones. However, at peak stress, in(peak sys) was able to separate three groups of segments. Receiver operating characteristic analysis showed an SR(peak sys) response of <0.5/s to identify patients with CAV with a sensitivity of 88%, specificity of 85% and a negative predictive value of 92%. CONCLUSION: Regional myocardial function is impaired in heart transplant recipients with CAV even when the disease is considered to be non-significant on conventional angiography. Systolic deformation parameters tended to detect the existence of CAV more accurately than conventional visual DSE assessment. Strain rate imaging during stress can therefore safely be used as a non-invasive screening test for detecting CAV in heart transplant recipients. PMID- 17916665 TI - Economic evaluation of ASCOT-BPLA: antihypertensive treatment with an amlodipine based regimen is cost effective compared with an atenolol-based regimen. AB - OBJECTIVE: To compare the cost effectiveness of an amlodipine-based strategy and an atenolol-based strategy in the treatment of hypertension in the UK and Sweden. DESIGN: A prospective, randomised trial complemented with a Markov model to assess long-term costs and health effects. SETTING: Primary care. PATIENTS: Patients with moderate hypertension and three or more additional risk factors. INTERVENTIONS: Amlodipine 5-10 mg with perindopril 4-8 mg added as needed or atenolol 50-100 mg with bendroflumethiazide 1.25-2.5 mg and potassium added as needed MAIN OUTCOME MEASURES: Cost per cardiovascular event and procedure avoided, and cost per quality-adjusted life-year gained. RESULTS: In the UK, the cost to avoid one cardiovascular event or procedure would be euro18 965, and the cost to gain one quality-adjusted life-year would be euro21 875. The corresponding figures for Sweden were euro13 210 and euro16 856. CONCLUSIONS: Compared with the thresholds applied by NICE and in the Swedish National Board of Health and Welfare's Guidelines for Cardiac Care, an amlodipine-based regimen is cost effective for the treatment of hypertension compared with an atenolol-based regimen in the population studied. PMID- 17916667 TI - Muscle metabolic, SR Ca(2+) -cycling responses to prolonged cycling, with and without glucose supplementation. AB - This study investigated the effects of prolonged exercise, with and without glucose supplementation, on metabolism and sarcoplasmic reticulum (SR) Ca(2+) handling properties in working vastus lateralis muscle. Fifteen untrained volunteers [peak O(2) consumption (Vo(2peak)) = 3.45 +/- 0.17 l/min; mean +/- SE] cycled at approximately 60% Vo(2peak) on two occasions, during which they were provided with either an artificially sweetened placebo beverage (NG) or a 6% glucose (G) beverage (~1.00 g carbohydrate/kg body mass). Beverage supplementation started at 30 min of exercise and continued every 15 min thereafter. SR Ca(2+) handling, metabolic, and substrate responses were assessed in tissue extracted from the vastus lateralis at rest, after 30 min and 90 min of exercise, and at fatigue in both conditions. Plasma glucose during G was 15-23% higher (P < 0.05) than those observed during NG following 60 min of exercise until fatigue. Cycle time to fatigue was increased (P < 0.05) by approximately 19% during G (137 +/- 7 min) compared with NG (115 +/- 6 min). Prolonged exercise reduced (P < 0.05) maximal Ca(2+)-ATPase activity (-18.4%), SR Ca(2+) uptake ( 27%), and both Phase 1 (-22.2%) and Phase 2 (-34.2%) Ca(2+)-release rates during NG. The exercise-induced reductions in SR Ca(2+)-cycling properties were not altered during G. The metabolic responses to exercise were all unaltered by glucose supplementation, since no differences in respiratory exchange ratios, carbohydrate and lipid oxidation rates, and muscle metabolite and glycogen contents were observed between NG and G. These results indicate that the maintenance of blood glucose homeostasis by glucose supplementation is without effect in modifying the muscle metabolic, endogenous glycogen, or SR Ca(2+) handling responses. PMID- 17916668 TI - Physiological role of free radicals in skeletal muscles. PMID- 17916669 TI - If I die before I wake: not a worry for sleep apnea patients. PMID- 17916670 TI - Muscle fiber type-specific response of Hsp70 expression in human quadriceps following acute isometric exercise. AB - To investigate the time course of fiber type-specific heat shock protein 70 (Hsp70) expression in human skeletal muscle after acute exercise, 10 untrained male volunteers performed single-legged isometric knee extensor exercise at 60% of their maximal voluntary contraction (MVC) with a 50% duty cycle (5-s contraction and 5-s relaxation) for 30 min. Muscle biopsies were collected from the vastus lateralis before (Pre) exercise in the rested control leg (C) and immediately after exercise (Post) in the exercised leg (E) only and on recovery days 1 (R1), 2 (R2), 3 (R3), and 6 (R6) from both legs. As demonstrated by Western blot analysis, whole muscle Hsp70 content was unchanged (P > 0.05) immediately after exercise (Pre vs. Post), was increased (P < 0.05) by approximately 43% at R1, and remained elevated throughout the entire recovery period in E only. Hsp70 expression was also assessed in individual muscle fiber types I, IIA, and IIAX/IIX by immunohistochemistry. There were no fiber type differences (P > 0.05) in basal Hsp70 expression. Immediately after exercise, Hsp70 expression was increased (P < 0.05) in type I fibers by approximately 87% but was unchanged (P > 0.05) in type II fibers (Pre vs. Post). At R1 and throughout recovery, Hsp70 content in E was increased above basal levels (P < 0.05) in all fiber types, but Hsp70 expression was always highest (P < 0.05) in type I fibers. Hsp70 content in C was not different from Pre at any time throughout recovery. Glycogen depletion was observed at Post in all type II, but not type I, fibers, suggesting that the fiber type differences in exercise induced Hsp70 expression were not related to glycogen availability. These results demonstrate that the time course of exercise-induced Hsp70 expression in human skeletal muscle is fiber type specific. PMID- 17916671 TI - Short-term treadmill running in the rat: what kind of stressor is it? AB - The use of short-term (1-5 days) treadmill running is becoming increasingly common as a model to study physiological adaptations following the exercise. Although the beneficial effects of acute exercise seem clear, a paucity of data exist describing potential markers of stress in response to forced running. We subjected male and female Sprague-Dawley rats to 0, 1, 2, 5, or 10 days of treadmill running. Twenty-four to 32 h after the last bout of exercise animals were killed and examined for training-induced changes in several physiological variables. No effect of skeletal citrate synthase activity was observed in the male animals after any duration, and only at 10 days of running did females show a significant increase in citrate synthase. Myocardial heat shock protein 72 (HSP72) content was higher in male rats than female rats, and exercise led to increased HSP72 in both sexes, although the time course was different between males and females. Animals displayed several markers of systemic stress in response to the treadmill running, and this was done in a sex-dependent manner. Serum corticosterone was significantly elevated in both sexes 24 h after exercise in three of four exercise groups. Corticosterone-binding globulin was higher in females, and decreased after running in female rats. Body and spleen weights decreased in males (but not females) in response to the exercise training, and running did not alter adrenal gland weights in either sex. These data indicate that in response to short-term treadmill running both male and female rats show signs of systemic stress, but that the pattern of changes occurs in a sex specific manner. PMID- 17916672 TI - Dexamethasone treatment of post-MI rats attenuates sympathetic innervation of the infarct region. AB - Sympathetic fiber innervation of the damaged region following injury represents a conserved event of wound healing. The present study tested the hypothesis that impaired scar healing in post-myocardial infarction (post-MI) rats was associated with a reduction of sympathetic fibers innervating the infarct region. In 1-wk post-MI rats, neurofilament-M-immunoreactive fibers (1,116 +/- 250 microm(2)/mm(2)) were detected innervating the infarct region and observed in close proximity to a modest number of endothelial nitric oxide synthase immunoreactive scar-residing vessels. Dexamethasone (Dex) treatment (6 days) of post-MI rats led to a significant reduction of scar weight (Dex + MI 38 +/- 4 mg vs. MI 63 +/- 2 mg) and a disproportionate nonsignificant decrease of scar surface area (Dex + MI 0.54 +/- 0.06 cm(2) vs. MI 0.68 +/- 0.06 cm(2)). In Dex treated post-MI rats, the density of neurofilament-M-immunoreactive fibers (125 +/- 47 microm(2)/mm(2)) innervating the infarct region was significantly reduced and associated with a decreased expression of nerve growth factor (NGF) mRNA (Dex + MI 0.80 +/- 0.07 vs. MI 1.11 +/- 0.08; P < 0.05 vs. MI). Previous studies have demonstrated that scar myofibroblasts synthesize NGF and may represent a cellular target of Dex. The exposure of 1st passage scar myofibroblasts to Dex led to a dose-dependent suppression of [(3)H]thymidine uptake and a concomitant attenuation of NGF mRNA expression (untreated 3.47 +/- 0.35 vs. Dex treated 2.28 +/- 0.40; P < 0.05 vs. untreated). Thus the present study has demonstrated that impaired scar healing in Dex-treated post-MI rats was associated with a reduction of neurofilament-M-immunoreactive fibers innervating the infarct region. The attenuation of scar myofibroblast proliferation and NGF mRNA expression may represent underlying mechanisms contributing to the diminished neural response in the infarct region of Dex-treated post-MI rats. PMID- 17916673 TI - Anaerobic and aerobic relative contribution to total energy release during supramaximal effort in patients with left ventricular dysfunction. AB - Energetic metabolism during effort is impaired in patients with left ventricular dysfunction (Dysf), but data have been lacking up to now on the relative anaerobic vs. aerobic contribution to total energy release during supramaximal effort. Recently, the maximal accumulated oxygen deficit (MAOD) has been shown to be measurable in Dysf patients, making it possible to evaluate the anaerobic/aerobic interaction under conditions of maximal stress of both anaerobic and aerobic metabolic pathways in this population. Nineteen Dysf patients and 17 normal patients (N) underwent one ramp cardiopulmonary, three moderate-intensity constant-power, and three supramaximal constant-power (1- to 2 min, 2- to 3-min, and 3- to 4-min duration) exercise tests. MAOD was the difference between accumulated O(2) demand (accO(2)dem; estimated from the moderate-intensity O(2) uptake/watt relationship) and uptake during supramaximal tests. Percent anaerobic (%Anaer) and aerobic (%Aer) energetic release were [(MAOD/accO(2)dem).100] and 100 - %Anaer, respectively. MAOD did not vary between 1-2, 2-3, and 3-4 min supramaximal tests, whereas accO(2)dem increased significantly with and was linearly related to test duration in both Dysf and N. Consequently, %Anaer and %Aer decreased and increased, respectively, with increasing test duration but did not differ between Dysf and N in 1-2 min, 2-3 min, and 3-4 min tests. Our study demonstrates a similar relative anaerobic vs. aerobic contribution to total energy release during supramaximal effort in Dysf and N. This finding indicates that energetic metabolism during supramaximal exercise is exercise tolerance independent and that relative anaerobic vs. aerobic contribution in this effort domain remains the same within the physiology or pathology-induced limits to individual peak exercise performance. PMID- 17916674 TI - Pulmonary C-fiber receptor activation abolishes uncoupled facial nerve activity from phrenic bursting during positive end-expired pressure in the rat. AB - Phasic respiratory bursting in the facial nerve (FN) can be uncoupled from phrenic bursting by application of 9 cmH(2)O positive end-expired pressure (PEEP). This response reflects excitation of expiratory-inspiratory (EI) and preinspiratory (Pre-I) facial neurons during the Pre-I period and inhibition of EI neurons during inspiration (I). Because activation of pulmonary C-fiber (PCF) receptors can inhibit the discharge of EI and Pre-I neurons, we hypothesized that PCF receptor activation via capsaicin would attenuate or abolish uncoupled FN bursting with an increase from 3 cmH(2)O (baseline) to 9 cmH(2)O PEEP. Neurograms were recorded in the FN and phrenic nerve in anesthetized, ventilated, vagally intact adult Wistar rats. Increasing PEEP to 9 cmH(2)O resulted in a persistent rhythmic discharge in the FN during phrenic quiescence (i.e., uncoupled bursting). Combination of PEEP with intrajugular capsaicin injection severely attenuated or eliminated uncoupled bursting in the FN (P < 0.05). Additional experiments examined the pattern of facial motoneuron (vs. neurogram) bursting during PEEP application and capsaicin treatment. These single-fiber recordings confirmed that Pre-I and EI (but not I) neurons continued to burst during PEEP induced phrenic apnea. Capsaicin treatment during PEEP substantially inhibited Pre-I and EI neuron discharge. Finally, analyses of FN and motoneuron bursting across the respiratory cycle indicated that the inhibitory effects of capsaicin were more pronounced during the Pre-I period. We conclude that activation of PCF receptors can inhibit FN bursting during PEEP-induced phrenic apnea by inhibiting EI and I facial motoneuron discharge. PMID- 17916676 TI - Strong galvanic vestibular stimulation obscures arterial pressure response to gravitational change in conscious rats. AB - Galvanic vestibular stimulation (GVS) is known to create an imbalance in the vestibular inputs; thus it is possible that the simultaneously applied GVS obscures adequate gravity-based inputs to the vestibular organs or modifies an input-output relationship of the vestibular system and then impairs the vestibular-mediated response. To examine this, arterial pressure (AP) response to gravitational change was examined in conscious rats with and without GVS. Free drop-induced microgravity and centrifugation-induced hypergravity were employed to elicit vestibular-mediated AP response. GVS itself induced pressor response in an intensity-dependent manner. This pressor response was completely abolished by vestibular lesion, suggesting that the GVS-induced response was mediated by the vestibular system. The pressor response to microgravity (35 +/- 3 mmHg) was significantly reduced by simultaneously applied GVS (19 +/- 1 mmHg), and pressor response to 3-G load was also significantly reduced by GVS. However, GVS had no effect on air jet-induced pressor response. The effects of GVS on pressor response to gravitational change were qualitatively and quantitatively similar to that caused by the vestibular lesion, effects of which were demonstrated in our previous studies (Gotoh TM, Fujiki N, Matsuda T, Gao S, Morita H. Am J Physiol Regul Integr Comp Physiol 286: R25-R30, 2004; Matsuda T, Gotoh TM, Tanaka K, Gao S, Morita H. Brain Res 1028: 140-147, 2004; Tanaka K, Gotoh TM, Awazu C, Morita H. Neurosci Lett 397: 40-43, 2006). These results indicate that GVS reduced the vestibular-mediated pressor response to gravitational change but has no effect on the non-vestibular-mediated pressor response. Thus GVS might be employed for the acute interruption of the AP response to gravitational change. PMID- 17916675 TI - TGF-beta1 and TNF-alpha are involved in the transcription of type I collagen alpha2 gene in soleus muscle atrophied by mechanical unloading. AB - The aim of this study was to examine the effect of hindlimb suspension (HS) on the expressions of COL1A2 (type I collagen alpha(2) chain) mRNA and its regulatory factors, transforming growth factors (TGF)-beta(1), -beta(2), and beta(3), phosphorylated Smad3, and tumor necrosis factor-alpha (TNF-alpha) in rat hindlimb muscles. Forty-eight male Wistar rats (age, 5 wk) were randomly assigned to HS for 1, 3, 7, and 14 days and control (n = 6 for each). During the exposure to HS, COL1A2 mRNA expression decreased in the soleus muscle at day 3 and recovered to control level at day 7. The content of TNF-alpha, one of the negative regulatory factors for COL1A2, increased from day 3 until day 14. On the other hand, the contents of TGF-beta(1), TGF-beta(3), and Smad3, positive regulatory factors for COL1A2, increased at day 7. The in situ hybridization for COL1A2 and the immunohistochemistry of TGF-beta(1) and TNF-alpha revealed their expressions around nerve-related tissues, including muscle spindles and connective tissue sheath. The results indicate that the transcriptional activity of COL1A2 in the soleus muscle initially decreases in response to unloading through an increase in TNF-alpha production; thereafter, it returns toward normal level through the activated TGF-beta/Smad pathway. PMID- 17916678 TI - Counterpoint: Chronic hypoxia-induced pulmonary hypertension does not lead to loss of pulmonary vasculature. PMID- 17916677 TI - Bleomycin-induced pulmonary fibrosis is attenuated by a monoclonal antibody targeting HER2. AB - The importance of HER2/HER3 signaling in decreasing the effects of lung injury was recently demonstrated. Transgenic mice unable to signal through HER2/HER3 had significantly less bleomycin-induced pulmonary fibrosis and showed a survival benefit. Based on these data, we hypothesized that pharmacological blockade of HER2/HER3 in vivo in wild-type mice would have the same beneficial effects. We tested this hypothesis in a bleomycin lung injury model using 2C4, a monoclonal antibody directed against HER2 that blocks HER2/HER3 signaling. The administration of 2C4 before injury decreased the effects of bleomycin at days 15 and 21 after injury. HER2/HER3 blockade resulted in less collagen deposition (362.8 +/- 37.9 compared with 610.5 +/- 27.1 microg/mg; P = 0.03) and less lung morphological changes (injury score of 1.99 +/- 1.55 vs. 3.90 +/- 0.76; P < 0.04). In addition, HER2/HER3 blockade resulted in a significant survival advantage with 50% vs. 25% survival at 30 days (P = 0.04). These results confirm that HER2 signaling can be pharmacologically targeted to reduce lung fibrosis and remodeling after injury. PMID- 17916681 TI - Chronic hypoxia-induced pulmonary hypertension does/does not lead to loss of pulmonary vasculature. PMID- 17916682 TI - Last Word on Point:Counterpoint "Chronic hypoxia-induced pulmonary hypertension does/does not lead to loss of pulmonary vasculature". PMID- 17916683 TI - Commentary on "The role of the large airways on smooth muscle contraction in asthma". PMID- 17916684 TI - Commentary on "The role of the large airways on smooth muscle contraction in asthma". PMID- 17916685 TI - Commentary on "The role of the large airways on smooth muscle contraction in asthma". PMID- 17916686 TI - Commentary on "The role of the large airways on smooth muscle contraction in asthma". PMID- 17916687 TI - Commentary on "The role of the large airways on smooth muscle contraction in asthma". PMID- 17916688 TI - Commentary on "The role of the large airways on smooth muscle contraction in asthma". PMID- 17916689 TI - Commentary on "The role of the large airways on smooth muscle contraction in asthma". PMID- 17916691 TI - Ordered phosphorylation governs oscillation of a three-protein circadian clock. AB - The simple circadian oscillator found in cyanobacteria can be reconstituted in vitro using three proteins-KaiA, KaiB, and KaiC. The total phosphorylation level of KaiC oscillates with a circadian period, but the mechanism underlying its sustained oscillation remains unclear. We have shown that four forms of KaiC differing in their phosphorylation state appear in an ordered pattern arising from the intrinsic autokinase and autophosphatase rates of KaiC and their modulation by KaiA. Kinetic and biochemical data indicate that one of these phosphoforms inhibits the activity of KaiA through interaction with KaiB, providing the crucial feedback that sustains oscillation. A mathematical model constrained by experimental data quantitatively reproduces the circadian period and the distinctive dynamics of the four phosphoforms. PMID- 17916692 TI - Telomeric repeat containing RNA and RNA surveillance factors at mammalian chromosome ends. AB - Telomeres, the DNA-protein complexes located at the end of linear eukaryotic chromosomes, are essential for chromosome stability. Until now, telomeres have been considered to be transcriptionally silent. We demonstrate that mammalian telomeres are transcribed into telomeric repeat-containing RNA (TERRA). TERRA molecules are heterogeneous in length, are transcribed from several subtelomeric loci toward chromosome ends, and localize to telomeres. We also show that suppressors with morphogenetic defects in genitalia (SMG) proteins, which are effectors of nonsense-mediated messenger RNA decay, are enriched at telomeres in vivo, negatively regulate TERRA association with chromatin, and protect chromosome ends from telomere loss. Thus, telomeres are actively transcribed into TERRA, and SMG factors represent a molecular link between TERRA regulation and the maintenance of telomere integrity. PMID- 17916693 TI - Video cameras on wild birds. AB - New Caledonian crows (Corvus moneduloides) are renowned for using tools for extractive foraging, but the ecological context of this unusual behavior is largely unknown. We developed miniaturized, animal-borne video cameras to record the undisturbed behavior and foraging ecology of wild, free-ranging crows. Our video recordings enabled an estimate of the species' natural foraging efficiency and revealed that tool use, and choice of tool materials, are more diverse than previously thought. Video tracking has potential for studying the behavior and ecology of many other bird species that are shy or live in inaccessible habitats. PMID- 17916695 TI - Sputnik nostalgia. PMID- 17916694 TI - Nanomechanical basis of selective gating by the nuclear pore complex. AB - The nuclear pore complex regulates cargo transport between the cytoplasm and the nucleus. We set out to correlate the governing biochemical interactions to the nanoscopic responses of the phenylalanineglycine (FG)-rich nucleoporin domains, which are involved in attenuating or promoting cargo translocation. We found that binding interactions with the transport receptor karyopherin-beta1 caused the FG domains of the human nucleoporin Nup153 to collapse into compact molecular conformations. This effect was reversed by the action of Ran guanosine triphosphate, which returned the FG domains into a polymer brush-like, entropic barrier conformation. Similar effects were observed in Xenopus oocyte nuclei in situ. Thus, the reversible collapse of the FG domains may play an important role in regulating nucleocytoplasmic transport. PMID- 17916696 TI - AIDS research. Promising AIDS vaccine's failure leaves field reeling. PMID- 17916697 TI - Human rights. Myanmar's secret history exposed in satellite images. PMID- 17916698 TI - Plant biology. At long last, pathologists hear plants' cry for help. PMID- 17916699 TI - Paleoanthropology. Nariokotome boy to go on the road despite protests. PMID- 17916700 TI - Climate change. Is battered Arctic Sea ice down for the count? PMID- 17916702 TI - Astronomy. Europeans lay down their wish list for next 2 decades. PMID- 17916701 TI - European science by the numbers. PMID- 17916703 TI - Sustainable science. Greening the meeting. PMID- 17916704 TI - Sustainability. Offsets: worth the price of emission? PMID- 17916705 TI - Sustainability. This man wants to green your lab. PMID- 17916707 TI - Sustainability. Do-it-yourself recycling. PMID- 17916706 TI - Sustainability. Energy-efficient freezers for everyone. PMID- 17916709 TI - Pseudoscience in Bosnia. PMID- 17916708 TI - A world with corals: what will it take? PMID- 17916710 TI - Effect of poor census data on population maps. PMID- 17916711 TI - Light-splitting method not new. PMID- 17916713 TI - Sustainability. Learning from 10 years of climate outlook forums in Africa. PMID- 17916712 TI - Comment on "Top-down versus bottom-up control of attention in the prefrontal and posterior parietal cortices". AB - Buschman and Miller (Reports, 30 March 2007, p. 1860) described the activity of ensembles of neurons in parietal and frontal cortex of monkeys performing visual search for targets that were easy or hard to distinguish from distractors. However, their conclusions are called into question by discrepancies between their results and publications from other laboratories measuring the same neural process. PMID- 17916714 TI - History of science. Sputnik and the Soviets. PMID- 17916715 TI - HISTORY OF SCIENCE: Science and Sputnik. AB - Sputnik led to an energetic U.S. space program and new discoveries about the cosmos. PMID- 17916716 TI - History of science. Sputnik and satellite astronomy. PMID- 17916717 TI - Evolution. Feathers, females, and fathers. PMID- 17916718 TI - Medicine. Testing hypotheses about autism. PMID- 17916719 TI - Biophysics. Going with the flow. PMID- 17916720 TI - Materials science. There's room in the middle. PMID- 17916722 TI - Life with oxygen. AB - The survival of all metazoan organisms is dependent on the regulation of O2 delivery and utilization to maintain a balance between the generation of energy and production of potentially toxic oxidants. Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that functions as a master regulator of oxygen homeostasis and has essential roles in metazoan development, physiology, and disease pathogenesis. Remarkable progress has been made in delineating the molecular mechanisms whereby changes in cellular oxygenation are transduced to the nucleus as changes in gene transcription through the activity of HIF-1. Pharmacologic agents that activate or inhibit the hypoxia signal transduction pathway may be useful therapies for ischemic and neoplastic disorders, respectively, which are the major causes of mortality in industrialized societies. PMID- 17916723 TI - PI3Kgamma is a key regulator of inflammatory responses and cardiovascular homeostasis. AB - Class I phosphoinositide 3-kinase (PI3K) signaling pathways regulate several important cellular functions, including cellular growth, division, survival, and movement. Class IB PI3K (also known as PI3Kgamma) links heterotrimeric GTP binding protein-coupled receptors to these pathways. Activation of class IB PI3K results in the rapid synthesis of phosphatidylinositol-3,4,5-trisphosphate [PtdIns(3,4,5)P3] and its dephosphorylation product PtdIns(3,4)P2 in the plasma membrane. These two lipid messengers bind to pleckstrin homology domain containing effectors that regulate a complex signaling web downstream of receptor activation. Characteristic features of this pathway are the regulation of protein kinases and the regulation of small guanosine triphosphatases that control cellular movement, adhesion, contraction, and secretion. Most of the ligands that activate class IB PI3K are involved in coordinating the body's response to injury and infection, and recent studies suggest that small molecule inhibitors of this enzyme may represent a novel class of anti-inflammatory therapeutic agents. PMID- 17916724 TI - Deconstructing the hedgehog pathway in development and disease. AB - The Hedgehog (Hh) family of secreted signaling proteins is a master regulator of cell fate determination in metazoans, contributing to both pattern formation during embryonic development and postembryonic tissue homeostasis. In a universally used mode of action, graded distribution of Hh protein induces differential cell fate in a dose-dependent manner in cells that receive Hh. Though much of this pathway has been elucidated from genetically based studies in model organisms, such as Drosophila and mice, the importance of Hh-mediated signaling in humans is clearly evident from malformations and a broad range of cancers that arise when the pathway is corrupted. PMID- 17916725 TI - Advances in cytokinin signaling. AB - Cytokinins are essential plant hormones that control various processes in plants' development and response to external stimuli. The Arabidopsis cytokinin signal transduction pathway involves hybrid histidine protein kinase sensors, phosphotransfer proteins, and regulators as transcription activators and repressors in a phosphorelay system. Each step is executed by components encoded by multigene families. Recent findings have revealed new functions, new feedback loops, and connections to other signaling pathways. PMID- 17916727 TI - Polymer gate dielectric surface viscoelasticity modulates pentacene transistor performance. AB - Nanoscopically confined polymer films are known to exhibit substantially depressed glass transition temperatures (Lg's) as compared to the corresponding bulk materials. We report here that pentacene thin films grown on polymer gate dielectrics at temperatures well below their bulk Tg's exhibit distinctive and abrupt morphological and microstructural transitions and thin-film transistor (TFT) performance discontinuities at well-defined growth temperatures. The changes reflect the higher chain mobility of the dielectric in its rubbery state and are independent of dielectric film thickness. Optimization of organic TFT performance must recognize this fundamental buried interface viscoelasticity effect, which is detectable in the current-voltage response. PMID- 17916726 TI - Odor-mediated push-pull pollination in cycads. AB - The reproductive organs of some plants self-heat, release scent, and attract pollinators. The relations among these processes are not well understood, especially in the more ancient, nonflowering gymnosperm lineages. We describe the influence of plant volatiles in an obligate pollination mutualism between an Australian Macrozamia cycad (a gymnosperm with male and female individuals) and its specialist thrips pollinator, Cycadothrips chadwicki. Pollen-laden thrips leave male cycad cones en masse during the daily thermogenic phase, when cone temperatures and volatile emissions increase dramatically and thrips are repelled. As thermogenesis declines, total volatile emissions diminish and cones attract thrips, resulting in pollination of female cones. Behavioral and electrophysiological tests on thrips reveal that variations in b-myrcene and ocimene emissions by male and female cones are sufficient to explain the observed sequential thrips' repellence (push) and attraction (pull). These dynamic interactions represent complex adaptations that enhance the likelihood of pollination and may reflect an intermediate state in the evolution of biotic pollination. PMID- 17916728 TI - Ultrastrong and stiff layered polymer nanocomposites. AB - Nanoscale building blocks are individually exceptionally strong because they are close to ideal, defect-free materials. It is, however, difficult to retain the ideal properties in macroscale composites. Bottom-up assembly of a clay/polymer nanocomposite allowed for the preparation of a homogeneous, optically transparent material with planar orientation of the alumosilicate nanosheets. The stiffness and tensile strength of these multilayer composites are one order of magnitude greater than those of analogous nanocomposites at a processing temperature that is much lower than those of ceramic or polymer materials with similar characteristics. A high level of ordering of the nanoscale building blocks, combined with dense covalent and hydrogen bonding and stiffening of the polymer chains, leads to highly effective load transfer between nanosheets and the polymer. PMID- 17916729 TI - Major Australian-Antarctic plate reorganization at Hawaiian-Emperor bend time. AB - A marked bend in the Hawaiian-Emperor seamount chain supposedly resulted from a recent major reorganization of the plate-mantle system there 50 million years ago. Although alternative mantle-driven and plate-shifting hypotheses have been proposed, no contemporaneous circum-Pacific plate events have been identified. We report reconstructions for Australia and Antarctica that reveal a major plate reorganization between 50 and 53 million years ago. Revised Pacific Ocean sea floor reconstructions suggest that subduction of the Pacific-Izanagi spreading ridge and subsequent Marianas/Tonga-Kermadec subduction initiation may have been the ultimate causes of these events. Thus, these plate reconstructions solve long standing continental fit problems and improve constraints on the motion between East and West Antarctica and global plate circuit closure. PMID- 17916730 TI - Absence of cooling in New Zealand and the adjacent ocean during the Younger Dryas chronozone. AB - As the climate warmed at the end of the last glacial period, a rapid reversal in temperature, the Younger Dryas (YD) event, briefly returned much of the North Atlantic region to near full-glacial conditions. The event was associated with climate reversals in many other areas of the Northern Hemisphere and also with warming over and near Antarctica. However, the expression of the YD in the mid- to low latitudes of the Southern Hemisphere (and the southwest Pacific region in particular) is much more controversial. Here we show that the Waiho Loop advance of the Franz Josef Glacier in New Zealand was not a YD event, as previously thought, and that the adjacent ocean warmed throughout the YD. PMID- 17916731 TI - A Cretaceous scleractinian coral with a calcitic skeleton. AB - It has been generally thought that scleractinian corals form purely aragonitic skeletons. We show that a well-preserved fossil coral, Coelosmilia sp. from the Upper Cretaceous (about 70 million years ago), has preserved skeletal structural features identical to those observed in present-day scleractinians. However, the skeleton of Coelosmilia sp. is entirely calcitic. Its fine-scale structure and chemistry indicate that the calcite is primary and did not form from the diagenetic alteration of aragonite. This result implies that corals, like other groups of marine, calcium carbonate-producing organisms, can form skeletons of different carbonate polymorphs. PMID- 17916732 TI - Sex chromosome-linked species recognition and evolution of reproductive isolation in flycatchers. AB - Interbreeding between species (hybridization) typically produces unfit offspring. Reduced hybridization should therefore be favored by natural selection. However, this is difficult to accomplish because hybridization also sets the stage for genetic recombination to dissociate species-specific traits from the preferences for them. Here we show that this association is maintained by physical linkage (on the same chromosome) in two hybridizing Ficedula flycatchers. By analyzing the mating patterns of female hybrids and cross-fostered offspring, we demonstrate that species recognition is inherited on the Z chromosome, which is also the known location of species-specific male plumage traits and genes causing low hybrid fitness. Limited recombination on the Z chromosome maintains associations of Z-linked genes despite hybridization, suggesting that the sex chromosomes may be a hotspot for adaptive speciation. PMID- 17916733 TI - Microbial population structures in the deep marine biosphere. AB - The analytical power of environmental DNA sequences for modeling microbial ecosystems depends on accurate assessments of population structure, including diversity (richness) and relative abundance (evenness). We investigated both aspects of population structure for microbial communities at two neighboring hydrothermal vents by examining the sequences of more than 900,000 microbial small-subunit ribosomal RNA amplicons. The two vent communities have different population structures that reflect local geochemical regimes. Descriptions of archaeal diversity were nearly exhaustive, but despite collecting an unparalleled number of sequences, statistical analyses indicated additional bacterial diversity at every taxonomic level. We predict that hundreds of thousands of sequences will be necessary to capture the vast diversity of microbial communities, and that different patterns of evenness for both high- and low abundance taxa may be important in defining microbial ecosystem dynamics. PMID- 17916734 TI - Genetic effects of captive breeding cause a rapid, cumulative fitness decline in the wild. AB - Captive breeding is used to supplement populations of many species that are declining in the wild. The suitability of and long-term species survival from such programs remain largely untested, however. We measured lifetime reproductive success of the first two generations of steelhead trout that were reared in captivity and bred in the wild after they were released. By reconstructing a three-generation pedigree with microsatellite markers, we show that genetic effects of domestication reduce subsequent reproductive capabilities by approximately 40% per captive-reared generation when fish are moved to natural environments. These results suggest that even a few generations of domestication may have negative effects on natural reproduction in the wild and that the repeated use of captive-reared parents to supplement wild populations should be carefully reconsidered. PMID- 17916736 TI - Chimpanzees are rational maximizers in an ultimatum game. AB - Traditional models of economic decision-making assume that people are self interested rational maximizers. Empirical research has demonstrated, however, that people will take into account the interests of others and are sensitive to norms of cooperation and fairness. In one of the most robust tests of this finding, the ultimatum game, individuals will reject a proposed division of a monetary windfall, at a cost to themselves, if they perceive it as unfair. Here we show that in an ultimatum game, humans' closest living relatives, chimpanzees (Pan troglodytes), are rational maximizers and are not sensitive to fairness. These results support the hypothesis that other-regarding preferences and aversion to inequitable outcomes, which play key roles in human social organization, distinguish us from our closest living relatives. PMID- 17916735 TI - Glia promote local synaptogenesis through UNC-6 (netrin) signaling in C. elegans. AB - Neural circuits are assembled through the coordinated innervation of pre- and postsynaptic partners. We show that connectivity between two interneurons, AIY and RIA, in Caenorhabditis elegans is orchestrated by a pair of glial cells that express UNC-6 (netrin). In the postsynaptic neuron RIA, the netrin receptor UNC 40 (DCC, deleted in colorectal cancer) plays a conventional guidance role, directing outgrowth of the RIA process ventrally toward the glia. In the presynaptic neuron AIY, UNC-40 (DCC) plays an unexpected and previously uncharacterized role: It cell-autonomously promotes assembly of presynaptic terminals in the immediate vicinity of the glial cell endfeet. These results indicate that netrin can be used both for guidance and local synaptogenesis and suggest that glial cells can function as guideposts during the assembly of neural circuits in vivo. PMID- 17916737 TI - Widespread role for the flowering-time regulators FCA and FPA in RNA-mediated chromatin silencing. AB - The RRM-domain proteins FCA and FPA have previously been characterized as flowering-time regulators in Arabidopsis. We show that they are required for RNA mediated chromatin silencing of a range of loci in the genome. At some target loci, FCA and FPA promote asymmetric DNA methylation, whereas at others they function in parallel to DNA methylation. Female gametophytic development and early embryonic development are particularly susceptible to malfunctions in FCA and FPA. We propose that FCA and FPA regulate chromatin silencing of single and low-copy genes and interact in a locus-dependent manner with the canonical small interfering RNA-directed DNA methylation pathway to regulate common targets. PMID- 17916738 TI - Methyl salicylate is a critical mobile signal for plant systemic acquired resistance. AB - In plants, the mobile signal for systemic acquired resistance (SAR), an organism wide state of enhanced defense to subsequent infections, has been elusive. By stimulating immune responses in mosaic tobacco plants created by grafting different genetic backgrounds, we showed that the methyl salicylate (MeSA) esterase activity of salicylic acid-binding protein 2 (SABP2), which converts MeSA into salicylic acid (SA), is required for SAR signal perception in systemic tissue, the tissue that does not receive the primary (initial) infection. Moreover, in plants expressing mutant SABP2 with unregulated MeSA esterase activity in SAR signal-generating, primary infected leaves, SAR was compromised and the associated increase in MeSA levels was suppressed in primary infected leaves, their phloem exudates, and systemic leaves. SAR was also blocked when SA methyl transferase (which converts SA to MeSA) was silenced in primary infected leaves, and MeSA treatment of lower leaves induced SAR in upper untreated leaves. Therefore, we conclude that MeSA is a SAR signal in tobacco. PMID- 17916739 TI - In situ imaging of the endogenous CD8 T cell response to infection. AB - Mounting a protective immune response is critically dependent on the orchestrated movement of cells within lymphoid organs. We report here the visualization, using major histocompatability complex class I tetramers, of the CD8-positive (CD8) T cell response in the spleens of mice to Listeria monocytogenes infection. A multistage pathway was revealed that included initial activation at the borders of the B and T cell zones followed by cluster formation with antigenpresenting cells leading to CD8 T cell exit to the red pulp via bridging channels. Strikingly, many memory CD8 T cells localized to the B cell zones and, when challenged, underwent rapid migration to the T cell zones where proliferation occurred, followed by egress via bridging channels in parallel with the primary response. Thus, the ability to track endogenous immune responses has uncovered both distinct and overlapping mechanisms and anatomical locations driving primary and secondary immune responses. PMID- 17916740 TI - Extranuclear steroid receptors: nature and actions. AB - Rapid effects of steroid hormones result from the actions of specific receptors localized most often to the plasma membrane. Fast-acting membrane-initiated steroid signaling (MISS) leads to the modification of existing proteins and cell behaviors. Rapid steroid-triggered signaling through calcium, amine release, and kinase activation also impacts the regulation of gene expression by steroids, sometimes requiring integration with nuclear steroid receptor function. In this and other ways, the integration of all steroid actions in the cell coordinates outcomes such as cell fate, proliferation, differentiation, and migration. The nature of the receptors is of intense interest, and significant data suggest that extranuclear and nuclear steroid receptor pools are the same proteins. Insights regarding the structural determinants for membrane localization and function, as well as the nature of interactions with G proteins and other signaling molecules in confined areas of the membrane, have led to a fuller understanding of how steroid receptors effect rapid actions. Increasingly, the relevance of rapid signaling for the in vivo functions of steroid hormones has been established. Examples include steroid effects on reproductive organ development and function, cardiovascular responsiveness, and cancer biology. However, although great strides have been made, much remains to be understood concerning the integration of extranuclear and nuclear receptor functions to organ biology. In this review, we highlight the significant progress that has been made in these areas. PMID- 17916741 TI - Uteroglobin: a steroid-inducible immunomodulatory protein that founded the Secretoglobin superfamily. AB - Blastokinin or uteroglobin (UG) is a steroid-inducible, evolutionarily conserved, secreted protein that has been extensively studied from the standpoint of its structure and molecular biology. However, the physiological function(s) of UG still remains elusive. Isolated from the uterus of rabbits during early pregnancy, UG is the founding member of a growing superfamily of proteins called Secretoglobin (Scgb). Numerous studies demonstrated that UG is a multifunctional protein with antiinflammatory/ immunomodulatory properties. It inhibits soluble phospholipase A(2) activity and binds and perhaps sequesters hydrophobic ligands such as progesterone, retinols, polychlorinated biphenyls, phospholipids, and prostaglandins. In addition to its antiinflammatory activities, UG manifests antichemotactic, antiallergic, antitumorigenic, and embryonic growth-stimulatory activities. The tissue-specific expression of the UG gene is regulated by several steroid hormones, although a nonsteroid hormone, prolactin, further augments its expression in the uterus. The mucosal epithelia of virtually all organs that communicate with the external environment express UG, and it is present in the blood, urine, and other body fluids. Although the physiological functions of this protein are still under investigation, a single nucleotide polymorphism in the UG gene appears to be associated with several inflammatory/autoimmune diseases. Investigations with UG-knockout mice revealed that the absence of this protein leads to phenotypes that suggest its critical homeostatic role(s) against oxidative damage, inflammation, autoimmunity, and cancer. Recent studies on UG binding proteins (receptors) provide further insight into the multifunctional nature of this protein. Based on its antiinflammatory and antiallergic properties, UG is a potential drug target. PMID- 17916742 TI - Neither DNA hypomethylation nor changes in the kinetics of erythroid differentiation explain 5-azacytidine's ability to induce human fetal hemoglobin. AB - 5-azacytidine (5-Aza) is a potent inducer of fetal hemoglobin (HbF) in people with beta-thalassemia and sickle cell disease. Two models have been proposed to explain this activity. The first is based on the drug's ability to inhibit global DNA methylation, including the fetal globin genes, resulting in their activation. The second is based on 5-Aza's cytotoxicity and observations that HbF production is enhanced during marrow recovery. We tested these models using human primary cells in an in vitro erythroid differentiation system. We found that doses of 5 Aza that produce near maximal induction of gamma-globin mRNA and HbF do not alter cell growth, differentiation kinetics, or cell cycle, but do cause a localized demethylation of the gamma promoter. However, when we reduced gamma promoter methylation to levels equivalent to those seen with 5-Aza or to the lower levels seen in primary fetal erythroid cells using DNMT1 siRNA and shRNA, we observed no induction of gamma-globin mRNA or HbF. These results suggest that 5-Aza induction of HbF is not the result of global DNA demethylation or of changes in differentiation kinetics, but involves an alternative, previously unrecognized mechanism. Other results suggest that posttranscriptional regulation plays an important role in the 5-Aza response. PMID- 17916743 TI - The impact of regulatory T cells on T-cell immunity following hematopoietic cell transplantation. AB - Regulatory T cells (Tregs) prevent graft-versus-host disease (GvHD) by inhibiting the proliferation and function of conventional T cells (Tcons). However, the impact of Tregs on T-cell development and immunity following hematopoietic cell transplantation (HCT) is unknown. Using a murine GvHD model induced by Tcons, we demonstrate that adoptive transfer of Tregs leads to (1) abrogration of GvHD, (2) preservation of thymic and peripheral lymph node architecture, and (3) an accelerated donor lymphoid reconstitution of a diverse TCR-Vbeta repertoire. The resultant enhanced lymphoid reconstitution in Treg recipients protects them from lethal cytomegalovirus (MCMV) infection. By contrast, mice that receive Tcons alone have disrupted lymphoid organs from GvHD and remain lymphopenic with a restricted TCR-Vbeta repertoire and rapid death on MCMV challenge. Lymphocytes from previously infected Treg recipients generate secondary response specific to MCMV, indicating long-term protective immunity with transferred Tregs. Thymectomy significantly reduces survival after MCMV challenge in Treg recipients compared with euthymic controls. Our results indicate that Tregs enhance immune reconstitution by preventing GvHD-induced damage of the thymic and secondary lymphoid microenvironment. These findings provide new insights into the role of Tregs in affording protection to lymphoid stromal elements important for T-cell immunity. PMID- 17916745 TI - The factor VIII C1 domain contributes to platelet binding. AB - Activated factor VIII (FVIIIa) forms a procoagulant complex with factor IXa on negatively charged membranes, including activated platelet surfaces. Membrane attachment involves the FVIII C2 domain; involvement of the adjacent C1 domain has not been established. Binding of recombinant FVIII C1C2 and C2 proteins to platelets was detected by flow cytometry using (1) anti-C2 monoclonal antibody ESH8 followed by a phycoerythrin-labeled secondary antibody; (2) biotinylated C1C2 detected by phycoerythrin-labeled streptavidin, and (3) C1C2 and C2 site specifically labeled with fluorescein. Highest binding and lowest background were obtained using fluorescein-conjugated proteins. More than 90% of activated platelets bound C1C2, compared with approximately 50% for equimolar C2. Estimates using fluorescent microbeads indicated approximately 7,000 C1C2-binding sites per platelet, approximately 1,400 for C2, and approximately 3,000 for fluorescein labeled FVIIIa. Unlike C2 or FVIII(a), C1C2 bound to approximately 700 sites/platelet before activation. C1C2 binding to activated platelets appeared independent of von Willebrand factor and was competed effectively by FVIII(a), but only partially by excess C2. Fluorescein-labeled FVIIIa was competed much more effectively by C1C2 than C2 for binding to activated platelets. Two monoclonal antibodies that inhibit C2 binding to membranes competed platelet binding of C2 more effectively than C1C2. Thus, the C1 domain of FVIII contributes to platelet-binding affinity. PMID- 17916746 TI - Imaging in staging of malignant lymphoma: a systematic review. AB - Computed tomography (CT) is currently the most commonly used means for staging malignant lymphoma. 18F-fluoro-2-deoxyglucose positron emission tomography (FDG PET), FDG-PET/CT fusion, and whole-body magnetic resonance imaging (WB-MRI) are potential alternatives. The purpose of this study was to systematically review published data on the diagnostic performance of CT, FDG-PET, FDG-PET/CT fusion, and WB-MRI in staging of malignant lymphoma. In addition, technical aspects, procedures, advantages, and drawbacks of each imaging modality are outlined. Three CT studies, 17 FDG-PET studies, and 4 FDG-PET/CT fusion studies were included in this systematic review. The studies were of moderate methodological quality and used different scoring systems to stage malignant lymphoma. CT remains the standard imaging modality for initial staging of malignant lymphoma, while FDG-PET has an essential role in restaging after treatment. Early results suggest that FDG-PET/CT fusion outperforms both CT alone and FDG-PET alone. Data on the diagnostic performance of WB-MRI are lacking. Future well-designed studies, expressing their results according to the Ann Arbor staging system, are needed to determine which imaging modality is most accurate and cost-effective in staging malignant lymphoma. PMID- 17916744 TI - Outcomes after allogeneic hematopoietic cell transplantation with nonmyeloablative or myeloablative conditioning regimens for treatment of lymphoma and chronic lymphocytic leukemia. AB - Allogeneic conventional hematopoietic cell transplantation (HCT) can be curative treatment for lymphoid malignancies, but it has been characterized by high nonrelapse mortality (NRM). Here, we compared outcomes among patients with lymphoma or chronic lymphocytic leukemia given either nonmyeloablative (n = 152) or myeloablative (n = 68) conditioning. Outcomes were stratified by the HCT specific comorbidity index. Patients in the nonmyeloablative group were older, had more previous treatment and more comorbidities, more frequently had unrelated donors, and more often had malignancy in remission compared with patients in the myeloablative group. Patients with indolent versus aggressive malignancies were equally distributed among both cohorts. After HCT, patients without comorbidities both in the nonmyeloablative and myeloablative cohorts had comparable NRM (P = .74), overall survival (P = .75), and progression-free survival (P = .40). No significant differences were observed (P = .91, P = .89, and P = .40, respectively) after adjustment for pretransplantation variables. Patients with comorbidities experienced lower NRM (P = .009) and better survival (P = .04) after nonmyeloablative conditioning. These differences became more significant (P < .001 and .007, respectively) after adjustment for other variables. Further, nonmyeloablative patients with comorbidities had favorable adjusted progression free survival (P = .01). Patients without comorbidities could be enrolled in prospective randomized studies comparing different conditioning intensities. Younger patients with comorbidities might benefit from reduced conditioning intensity. PMID- 17916747 TI - Activator-specific requirement of properdin in the initiation and amplification of the alternative pathway complement. AB - Properdin is a positive regulator of alternative pathway (AP) complement. The current understanding of properdin function is that it facilitates AP complement activation by stabilizing the C3 convertase C3bBb. Properdin-deficient patients are susceptible to lethal meningococcal infection, but the mechanism of this selective predisposition is not fully understood. By gene targeting in the mouse, we show here that properdin is essential for AP complement activation induced by bacterial lipopolysacharride (LPS) and lipooligosacharride (LOS) and other, but not all, AP complement activators. LPS- and LOS-induced AP complement activation was abolished in properdin-/- mouse serum, and properdin-/- mice were unable to clear Crry-deficient erythrocytes, which are known to be susceptible to AP complement-mediated extravascular hemolysis. In contrast, zymosan- and cobra venom factor-induced AP complement activation, and classical pathway-triggered AP complement amplification were only partially or minimally affected in properdin-/ mice. We further show that the ability of human properdin to restore LPS dependent AP complement activity in properdin-/- mouse serum correlated with the human properdin-binding affinity of the LPS. These results reveal a novel role of properdin in AP complement initiation and have implications for understanding the selective predisposition of properdin-deficient patients to meningococcal infection. PMID- 17916748 TI - M-CSF: a novel plasmacytoid and conventional dendritic cell poietin. AB - The critical importance of plasmacytoid dendritic cells (pDCs) in viral infection, autoimmunity, and tolerance has focused major attention on these cells that are rare in blood and immune organs of humans and mice. The recent development of an Flt-3 ligand (FL) culture system of bone marrow cells has led to the simple generation of large numbers of pDCs that resemble their in vivo steady-state counterparts. The FL system has allowed unforeseen insight into the biology of pDCs, and it is assumed that FL is the crucial growth factor for these cells. Surprisingly we have found that a cell type with high capacity for interferon-alpha (IFN-alpha) production in response to CpG-containing oligonucleotides, a feature of pDCs, develop within macrophage-colony-stimulating factor (M-CSF)-generated bone marrow cultures. Analysis of this phenomenon revealed that M-CSF is able to drive pDCs as well as conventional DCs (cDCs) from BM precursor cells in vitro. Furthermore, application of M-CSF to mice was able to drive pDCs and cDCs development in vivo. It is noteworthy that using mice deficient in FL indicated that the M-CSF-driven generation of pDCs and cDCs in vitro and in vivo was independent of endogenous FL. PMID- 17916751 TI - Extracellular localization of epimorphin/syntaxin-2. PMID- 17916753 TI - Prognostic impact of elevated pretransplantation serum ferritin in patients undergoing myeloablative stem-cell transplantation. PMID- 17916754 TI - Immunogenicity of factor VIII concentrates in patients with hemophilia: a randomized clinical trial is warranted. PMID- 17916749 TI - Small-molecule XIAP antagonist restores caspase-9 mediated apoptosis in XIAP positive diffuse large B-cell lymphoma cells. AB - Clinical outcome in patients with primary nodal diffuse large B-cell lymphomas (DLBCLs) is correlated with expression of inhibitors of the intrinsic apoptosis pathway, including X-linked inhibitor of apoptosis protein (XIAP). XIAP suppresses apoptosis through inhibiting active caspase-3, caspase-7, and caspase 9. In this study, we investigated to see if the small-molecule XIAP antagonist 1396-12 induces cell death in cultured lymphoma cells of patients with DLBCL. Treatment with this XIAP antagonist resulted in relief of caspase-3 inhibition and in induction of apoptosis in 16 of 20 tested DLBCL samples. Sensitivity to the XIAP antagonist was observed in both chemotherapy-refractory and -responsive DLBCL, but did not affect peripheral blood mononuclear cells and tonsil germinal center B cells from healthy donors. XIAP antagonist-sensitive samples were characterized by high expression levels of XIAP, relatively low expression levels of Bcl-2, and by constitutive caspase-9 activation. These data indicate that the small-molecule XIAP antagonist can induce apoptosis in cultured DLBCL cells and therefore should be considered for possible development as a therapy for these patients. In vitro sensitivity to the XIAP antagonist can be predicted based on biological markers, suggesting the possibility of predefining patients most likely to benefit from XIAP antagonist therapy. PMID- 17916755 TI - Chronic megakaryocytic leukemia, misnamed chronic idiopathic myelofibrosis, has neoplastic not hyperplastic megakaryocytopoiesis. PMID- 17916757 TI - Philadelphia-negative clonal hematopoiesis is a significant feature of dasatinib therapy for chronic myeloid leukemia. PMID- 17916758 TI - Modulation of nicotinamide adenine dinucleotide phosphate oxidase expression and function by 3',4'-dihydroxyflavonol in phagocytic and vascular cells. AB - Previously we have demonstrated that 3',4'-dihydroxyflavonol (DiOHF), a novel synthetic flavonol, protects against ischemia reperfusion injury in both heart and brain. In this study, we characterized the pharmacological effects of DiOHF on phagocytic and vascular NADPH oxidase. Superoxide release (lucigenin-enhanced chemiluminescence or cytochrome c reduction), NADPH oxidase activation (membrane translocation of p47phox), and subunit expression (real-time polymerase chain reaction and Western blot) were examined in differentiated HL-60 cells, human neutrophils, vascular endothelial and smooth muscle cells, and mouse aorta. DiOHF concentration dependently suppressed superoxide accumulation (EC(50) = 8.4 +/- 1.7 microM) in vascular smooth muscle cells, which appears to be attributable to its superoxide scavenging activity (EC(50) = 6.1 +/- 1.1 microM measured in a cell-free system). DiOHF had similar effects in HL-60 cells and isolated aortic rings. In HL-60 cells, but not endothelial or smooth muscle cells, DiOHF and quercetin (10 and 30 microM) significantly reduced the protein expression of p47phox, whereas p67phox was not altered. DiOHF did not affect phorbol ester induced membrane translocation of either p47phox or protein kinase C in leukocytes. Our results suggest that suppression of NADPH oxidase-dependent superoxide accumulation may contribute to the cytoprotective actions of DiOHF during ischemia-reperfusion injury. PMID- 17916759 TI - Protective effects of estradiol on ethanol-induced bone loss involve inhibition of reactive oxygen species generation in osteoblasts and downstream activation of the extracellular signal-regulated kinase/signal transducer and activator of transcription 3/receptor activator of nuclear factor-kappaB ligand signaling cascade. AB - Bone loss occurs following chronic ethanol (EtOH) consumption in males and cycling females in part as a result of increased bone resorption. We have demonstrated in vivo that estradiol treatment can reverse this effect. Using osteoclast precursors from bone marrow and osteoblast/preosteoclast coculture, we found that EtOH-induced receptor activator of nuclear factor-kappaB ligand (RANKL) expression in osteoblasts was able to promote osteoclastogenesis. These effects were blocked by pretreatment of cells with either 17beta-estradiol (E(2)) or the anti-oxidant N-acetyl cysteine (NAC). EtOH treatment of stromal osteoblasts increased the intracellular level of reactive oxygen species (ROS). This was associated with induction of NADPH oxidase (NOX) and a downstream signaling cascade involving sustained activation of extracellular signal regulated kinase (ERK) and activation of signal transducer and activator of transcription 3, resulting in increased gene expression of RANKL. In the presence of EtOH, sustained nuclear ERK translocation >24 h was observed in calvarial osteoblasts and UMR-106 cells transfected with green fluorescent protein-ERK2 plasmid. This was abolished by pretreatment with either E(2) or NAC. NOX subtypes 1, 2, and 4, but not 3, were expressed in stromal osteoblasts. Chemical inhibition of NOX by diphenylene iodonium also reversed the ability of EtOH to phosphorylate ERK and induce RANKL mRNA expression. Down-regulation of EtOH induced ROS generation in osteoblasts was also observed after treatment with E(2) or NAC. These data suggest that the molecular mechanisms whereby E(2) prevents EtOH-induced bone loss involve interference with ROS generation and cytoplasmic kinase activation. PMID- 17916760 TI - Pharmacological characterization of indacaterol, a novel once daily inhaled 2 adrenoceptor agonist, on small airways in human and rat precision-cut lung slices. AB - Indacaterol is a novel once daily inhaled beta(2) adrenoceptor agonist in clinical development. This study compared the properties of indacaterol with salmeterol, formoterol, and albuterol on small airways in precision-cut lung slices from human and rat contracted with carbachol and serotonin, respectively. In human lung slices, the rank order of potency was formoterol >/= salmeterol > indacaterol > albuterol, respectively. Indacaterol had similar intrinsic efficacy to formoterol, followed by albuterol and salmeterol. The onset of action was fast for albuterol, formoterol, and indacaterol, whereas it was significantly slower for salmeterol. The duration of action ranking was indacaterol > salmeterol > formoterol > albuterol. When compared with human lung slices, in the rat lung slices, similar potency, intrinsic efficacy, and onset of action were observed for indacaterol, formoterol, and salmeterol. Albuterol had an increased potency when compared with human lung slices and a slower onset of action. In conclusion, our results show that the human lung slice system seems to be a good model to study the clinical properties of inhaled long-acting beta(2) adrenoceptor agonists and that caution is needed extrapolating from rat model to humans. Finally, using the human lung slice model, we have characterized indacaterol as a fast acting compound with a longer duration of action than salmeterol and formoterol. PMID- 17916761 TI - Interleukin-6 174G/C polymorphism and ischemic stroke: a systematic review. AB - BACKGROUND AND PURPOSE: Interleukin-6 (IL-6) is associated with atherosclerotic disease and is also a key mediator in the inflammatory response to cerebral ischemia. Although the IL-6 -174G/C promoter polymorphism has been associated with carotid artery atherosclerosis and coronary heart disease, its relation to ischemic stroke is unclear. This review summarizes the current literature and discusses methodological considerations for future studies. METHODS: Electronic searches were conducted in the PubMed MEDLINE, Scopus, and ISI Web of Science databases. Two investigators independently reviewed all abstracts to identify studies examining the association between the IL-6 -174G/C polymorphism and ischemic cerebrovascular events. RESULTS: Twelve relevant publications were identified. Three reported on a subset of patients from a later publication, leaving 9 independent studies. Two studies found an association between ischemic stroke and the G allele or GG genotype, whereas 4 found an association with the C allele or CC genotype. One study found the CC genotype to be significantly less frequent in retinal artery occlusion patients. Two studies found no association between the -174G/C polymorphism and stroke. CONCLUSIONS: Studies investigating stroke and the -174G/C polymorphism report conflicting results, which may reflect the complex physiology of IL-6 and true differences between stroke subtypes and populations. However, interpretation of published results is hindered by methodological limitations, and greater rigor and consistency in future studies will help unravel the relationship between the -174G/C polymorphism and stroke. PMID- 17916763 TI - Development and validation of a short form of the Fugl-Meyer motor scale in patients with stroke. AB - BACKGROUND AND PURPOSE: The 50-item Fugl-Meyer motor scale (FM) is commonly used in outcome studies. However, the lengthy administration time of the FM keeps it from being widely accepted for routine clinical use. We aimed to develop a short form of the FM (the S-FM) with sound psychometric properties for stroke patients. METHODS: The FM was administered to 279 patients. It was then simplified based on expert opinions and the results of Rasch analysis. The psychometric properties (including Rasch reliability, concurrent validity, predictive validity, and responsiveness) of the S-FM were examined and were compared with those of the FM. The concurrent validity and responsiveness of the S-FM were further validated in a sample from the Netherlands. RESULTS: We selected 6 items for each subscale to construct a 12-item S-FM. The S-FM demonstrated high Rasch reliability, high concurrent validity with the original scale, moderate responsiveness, and moderate predictive validity with the comprehensive activities of daily living function. The S-FM also showed sufficient concurrent validity and responsiveness on the Dutch sample. CONCLUSIONS: Our results provide strong evidence that the psychometric properties of the S-FM are comparable with those of the FM. The S-FM contains only 12 items, making it a very efficient measure for assessing the motor function of stroke patients in both clinical and research settings. PMID- 17916765 TI - Novel end point analytic techniques and interpreting shifts across the entire range of outcome scales in acute stroke trials. AB - BACKGROUND AND PURPOSE: Stroke treatments are generally not curative, but rather alter patient outcome over the entire range of functional measures. Dichotomizing outcome scales reduces computational complexity, but discards substantial outcome information, artificially privileges only a single health state transition as clinically meaningful, and often reduces study power. Newer approaches to endpoint analysis have several advantageous properties. Summary of Review- The global statistic assesses treatment effects on multiple outcome measures simultaneously. However, translating the global statistic multidimensional vector effect at the population level into benefit or harm expected in the individual patient is problematic. Responder analysis adjusts outcome thresholds to patient stroke severity at study entry, identifying achievable goals for each patient. However, responder analysis still discards substantial outcome information. Shift analysis gauges change in outcome distributions over the full range of ascertained outcomes, incorporating benefit and harm at all health state transitions valued by patients and clinicians, and often increasing study power. Translation of findings of shift analyses into clinically accessible terms may be accomplished using the recently developed joint outcome table specification technique, which yields the following values for the number needed to treat for 1 patient to improve in a clinically important manner: nimodipine in subarachnoid hemorrhage, 6.8; coiling over clipping, 5.9; intra-arterial pro-urokinase in acute cerebral ischemia, 4.8; intravenous tissue plasminogen activator, 3.3. CONCLUSIONS: Dichotomized, global statistic, responder, and shift analyses each offer distinctive benefits and drawbacks. Choice of primary end point analytic technique should be tailored to the study population, expected treatment response, and study purpose. Shift analysis generally provides the most comprehensive index of a treatment's clinical impact. PMID- 17916764 TI - NADPH oxidase plays a central role in blood-brain barrier damage in experimental stroke. AB - BACKGROUND AND PURPOSE: Cerebral ischemia/reperfusion is associated with reactive oxygen species (ROS) generation, and NADPH oxidases are important sources of ROS. We hypothesized that NADPH oxidases mediate blood-brain barrier (BBB) disruption and contribute to tissue damage in ischemia/reperfusion. METHODS: Ischemia was induced by filament occlusion of the middle cerebral artery in mice for 2 hours followed by reperfusion. BBB permeability was measured by Evans blue extravasation. Monolayer permeability was determined from transendothelial electrical resistance of cultured porcine brain capillary endothelial cells. RESULTS: BBB permeability was increased in the ischemic hemisphere 1 hour after reperfusion. In NADPH oxidase-knockout (gp91phox(-/-)) mice, middle cerebral artery occlusion-induced BBB disruption and lesion volume were largely attenuated compared with those in wild-type mice. Inhibition of NADPH oxidase by apocynin prevented BBB damage. In porcine brain capillary endothelial cells, hypoxia/reoxygenation induced translocation of the NADPH oxidase activator Rac-1 to the membrane. In vivo inhibition of Rac-1 by the 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor atorvastatin or Clostridium difficile lethal toxin B also prevented the ischemia/reperfusion-induced BBB disruption. Stimulation of porcine brain capillary endothelial cells with H(2)O(2) increased permeability, an effect attenuated by inhibition of phosphatidyl inositol 3-kinase or c-Jun N terminal kinase but not blockade of extracellular signal-regulated kinase-1/2 or p38 mitogen-activated protein kinase. Inhibition of Rho kinase completely prevented the ROS-induced increase in permeability and the ROS-induced polymerization of the actin cytoskeleton. CONCLUSIONS: Activation of Rac and subsequently of the gp91phox containing NADPH oxidase promotes cerebral ROS formation, which then leads to Rho kinase-mediated endothelial cell contraction and disruption of the BBB. Inhibition of NADPH oxidase is a promising approach to reduce brain injury after stroke. PMID- 17916766 TI - Carnosine is neuroprotective against permanent focal cerebral ischemia in mice. AB - BACKGROUND AND PURPOSE: Carnosine is a naturally occurring dipeptide with multiple neuroprotective properties. In addition, it is well tolerated in high doses with minimal side effects. The purposes of this study were to determine whether carnosine is neuroprotective in permanent focal cerebral ischemia and to determine potential mechanisms of neuroprotection. METHODS: We investigated the efficacy of carnosine in a mouse model of permanent focal cerebral ischemia. The effects of carnosine were investigated with respect to neuronal damage and infarct formation, endogenous antioxidant status, and matrix metalloproteinase activity. RESULTS: Carnosine significantly decreased infarct size and neuronal damage when administered at time points both before and after the induction of ischemia. Carnosine also decreased reactive oxygen species levels in the ischemic brain, preserved normal glutathione levels, and decreased matrix metalloproteinase protein levels and activity. CONCLUSIONS: Carnosine is neuroprotective in focal cerebral ischemia and appears to influence deleterious pathological processes that are activated after the onset of ischemia. PMID- 17916767 TI - Stroke incidence, prognosis, 30-day, and 1-year case fatality rates in Matao, Brazil: a population-based prospective study. AB - BACKGROUND AND PURPOSE: Stroke is the main cause of death in Brazil and no prospective population-based study has been conducted in the country. The purpose of this study was to determine the incidence of stroke, stroke subtypes, case fatality, and prognosis after 1 year of follow-up in the city of Matao, Brazil. METHODS: Using overlapping methods of case ascertainment, all stroke cases that occurred in the city of Matao (population, 75,053) from November 1, 2003 to October 31, 2004 were followed-up at 1 month and 1 year after the episode. Standard criteria for population-based stroke studies were followed. RESULTS: Of 141 suspect stroke cases identified, 81 were first-ever-in-a-lifetime strokes. All patients underwent a CT scan. The crude annual incidence rate per 100,000 per year was 108 (95% CI, 85.7-134.1) and the rate adjusted for sex and age to the Segi population was 137 (95% CI, 112.0 to 166.4) per 100,000 inhabitants per year. Ischemic stroke occurred in 69 (85.2%) subjects, intracerebral hemorrhage in 11 (13.6%), and subarachnoid hemorrhage in 1 (1.2%). The 30-day case fatality rate was 18.5 and the 1-year case fatality rate was 30.9%. After 1 year of follow up, 43% of the patients were independent in activities of daily living, 49.4% had independent gait, and 15.9% had a recurrent stroke. CONCLUSIONS: The present results are similar to those obtained in other stroke population-based studies. Future studies in Matao will provide the opportunity to discover ways to decrease the stroke burden in Brazil. PMID- 17916768 TI - Reversed Robin Hood syndrome in acute ischemic stroke patients. AB - BACKGROUND AND PURPOSE: Recurrent hemodynamic and neurological changes with persisting arterial occlusions may be attributable to cerebral blood flow steal from ischemic to nonaffected brain. METHODS: Transcranial Doppler monitoring with voluntary breath-holding and serial NIH Stroke Scale (NIHSS) scores were obtained in patients with acute middle cerebral artery or internal carotid artery occlusions. The steal phenomenon was detected as transient, spontaneous, or vasodilatory stimuli-induced velocity reductions in affected arteries at the time of velocity increase in normal vessels. The steal magnitude (%) was calculated as [(MFVm-MFVb)/MFVb]x100, where m=minimum and b=baseline mean flow velocities (MFV) during the 15- to 30-second period of a total 30 second of breath-holding. RESULTS: Six patients had steal phenomenon on transcranial Doppler (53 to 73 years, NIHSS 4 to 15 points). Steal magnitude ranged from -15.0% to -43.2%. All patients also had recurrent neurological worsening (>2 points increase in NIHSS scores) at stable blood pressure. In 3 of 5 patients receiving noninvasive ventilatory correction for snoring/sleep apnea, no further velocity or NIHSS score changes were noted. CONCLUSIONS: Our descriptive study suggests possibility to detect and quantify the cerebral steal phenomenon in real-time. If the steal is confirmed as the cause of neurological worsening, reversed Robin Hood syndrome may identify a target group for testing blood pressure augmentation and noninvasive ventilatory correction in stroke patients. PMID- 17916769 TI - IGF-1 reduces inflammatory responses, suppresses oxidative stress, and decreases atherosclerosis progression in ApoE-deficient mice. AB - OBJECTIVE: Whereas growth factors, via their ability to stimulate vascular smooth muscle cell (VSMC) proliferation and migration, have been thought to play a permissive role in atherosclerosis initiation and progression, the role of insulin-like growth factor-1 (IGF-1) is unknown. Here we report for the first time that IGF-1 infusion decreased atherosclerotic plaque progression in ApoE deficient mice on a Western diet. METHODS AND RESULTS: ApoE-null mice (8 weeks) were infused with vehicle or recombinant human IGF-1 and fed a high-fat diet for 12 weeks. Analysis of aortic sinuses revealed that IGF-1 infusion decreased atherosclerotic plaque progression and macrophage infiltration into lesions. Furthermore, IGF-1 decreased vascular expression of the proinflammatory cytokines interleukin-6 and tumor necrosis factor-alpha, reduced aortic superoxide formation and urinary 8-isoprostane levels, and increased aortic pAkt and eNOS expression and circulating endothelial progenitor cells, consistent with an antiinflammatory, antioxidant, and prorepair effect on the vasculature. CONCLUSIONS: Our data indicate that an increase in circulating IGF-1 reduces vascular inflammatory responses, systemic and vascular oxidant stress and decreases atherosclerotic plaque progression. These findings have major implications for the treatment of atherosclerosis. PMID- 17916770 TI - Intrinsic pathway of coagulation and arterial thrombosis. AB - Formation of a fibrin clot is mediated by a group of tightly regulated plasma proteases and cofactors. While this system is essential for minimizing blood loss from an injured blood vessel (hemostasis), it also contributes to pathologic fibrin formation and platelet activation that may occlude vessels (thrombosis). Many antithrombotic drugs target key elements of the plasma coagulation mechanism such as thrombin and factor Xa, based on the premise that plasma elements contributing to thrombosis are primarily those involved in hemostasis. Recent studies with genetically altered mice raise questions about this paradigm. Deficiencies of the intrinsic pathway proteases factor XII and factor XI are not associated with abnormal hemostasis in mice, but impair formation of occlusive thrombi in arterial injury models, indicating that pathways not essential for hemostasis participate in arterial thrombosis. If factor XII or factor XI make similar contributions to thrombosis in humans, these proteases could be ideal targets for drugs to treat or prevent thromboembolic disease with minimal risk of therapy-associated bleeding. PMID- 17916771 TI - Rosiglitazone reduces glucose-induced oxidative stress mediated by NAD(P)H oxidase via AMPK-dependent mechanism. AB - OBJECTIVE: Hyperglycemia is the main determinant of long-term diabetic complications, mainly through induction of oxidative stress. NAD(P)H oxidase is a major source of glucose-induced oxidative stress. In this study, we tested the hypothesis that rosiglitazone (RSG) is able to quench oxidative stress initiated by high glucose through prevention of NAD(P)H oxidase activation. METHODS AND RESULTS: Intracellular ROS were measured using the fluoroprobe TEMPO-9-AC in HUVECs exposed to control (5 mmol/L) and moderately high (10 mmol/L) glucose concentrations. NAD(P)H oxidase and AMPK activities were determined by Western blot. We found that 10 mmol/L glucose increased significantly ROS production in comparison with 5 mmol/L glucose, and that this effect was completely abolished by RSG. Interestingly, inhibition of AMPK, but not PPARgamma, prevented this effect of RSG. AMPK phosphorylation by RSG was necessary for its ability to hamper NAD(P)H oxidase activation, which was indispensable for glucose-induced oxidative stress. Downstream of AMPK activation, RSG exerts antioxidative effects by inhibiting PKC. CONCLUSIONS: This study demonstrates that RSG activates AMPK which, in turn, prevents hyperactivity of NAD(P)H oxidase induced by high glucose, possibly through PKC inhibition. Therefore, RSG protects endothelial cells against glucose-induced oxidative stress with an AMPK-dependent and a PPARgamma-independent mechanism. PMID- 17916772 TI - Blocking thrombospondin-1/CD47 signaling alleviates deleterious effects of aging on tissue responses to ischemia. AB - OBJECTIVE: Decreased blood flow secondary to peripheral vascular disease underlies a significant number of chronic diseases that account for the majority of morbidity and mortality among the elderly. Blood vessel diameter and blood flow are limited by the matricellular protein thrombospondin-1 (TSP1) through its ability to block responses to the endogenous vasodilator nitric oxide (NO). In this study we investigate the role TSP1 plays in regulating blood flow in the presence of advanced age and atherosclerotic vascular disease. METHODS AND RESULTS: Mice lacking TSP1 or CD47 show minimal loss of their resistance to ischemic injury with age and increased preservation of tissue perfusion immediately after injury. Treatment of WT and apolipoprotein E-null mice using therapeutic agents that decrease CD47 or enhance NO levels reverses the deleterious effects of age- and diet-induced vasculopathy and results in significantly increased tissue survival in models of ischemia. CONCLUSIONS: With increasing age and diet-induced atherosclerotic vascular disease, TSP1 and its receptor CD47 become more limiting for blood flow and tissue survival after ischemic injury. Drugs that limit TSP1/CD47 regulation of blood flow could improve outcomes from surgical interventions in the elderly and ameliorate vascular complications attendant to aging. PMID- 17916774 TI - Mapping, genetic isolation, and characterization of genetic loci that determine resistance to atherosclerosis in C3H mice. AB - OBJECTIVE: C3H/HeJ (C3H) mice are extremely resistant to atherosclerosis. To identify the genetic factors involved in lesion initiation, we studied a cross between C3H and the susceptible strain C57BL/6J (B6) on a hyperlipidemic (apolipoprotein E-null) background. METHODS AND RESULTS: Whereas a previous cross in mice fed a Western diet for 16 weeks revealed a very complex inheritance pattern with many significant lesion QTLs, the present cross, on a chow diet, revealed a single major locus on chromosome 9 (lod=5.0, Ath29*), and a suggestive locus on chromosome 4 (lod=2.6, Ath8). QTLs for plasma HDL, total cholesterol, and triglyceride levels were found on chromosome 1 over the ApoA2 gene. Neither of the lesion QTLs were associated with differences in plasma lipid levels or other systemic risk factors, consistent with the concept that genetic factors affecting cellular functions of the vessel wall are important determinants of atherosclerosis susceptibility. We generated a congenic strain for Ath29 and confirmed its contribution to lesion development. Toll-like receptor 4 (Tlr4), the lipopolysaccharide (LPS) receptor, is located in the Ath8 region and is known to be defective in C3H/HeJ mice. We constructed a congenic strain carrying a normal Tlr4 gene on the C3H Apoe-null background and found that the defective Tlr4 does not contribute significantly to lesion resistance during early lesion development. CONCLUSIONS: We identified one major QTL on chromosome 9, Ath29, for early lesion development in the BXH ApoE(-/-) cross fed on a chow diet and confirmed its contribution in congenic mice. We have also determined that Tlr4 on the C3H ApoE(-/-) background does not contribute to early lesion development. *Ath29 is referred to as Ath22 in Su et al 2006. PMID- 17916773 TI - Statin treatment and 3' polyadenylation of eNOS mRNA. AB - OBJECTIVE: Statins have been shown to increase endothelial nitric oxide synthase expression via enhanced mRNA stability. Because the poly(A) tail is an important determinant of transcript stability, we sought to characterize the effect of statins on eNOS mRNA 3' polyadenylation. METHODS AND RESULTS: Endothelial cells treated with statins had a time- and dose-dependent increase in eNOS transcripts with long poly(A) tails (75 to 160 adenosines). This effect was dependent on 3 hydroxy-3-methylglutaryl (HMG)-coenxyme A (CoA) reductase inhibition and was observed with both lipophilic (simvastatin) and hydrophilic (rosuvastatin) statins. In mRNA stability assays, polyadenylated eNOS transcripts from statin treated cells were 2- to 3-fold more stable than transcripts from untreated cells. The effect of statins on eNOS polyadenylation was related to cytoskeleton organization; there was increased eNOS mRNA polyadenylation after Rho inhibition and cytochalasin D treatment. Further, we found increased phosphorylation of RNA polymerase II in statin-treated cells, suggesting that statin-induced polyadenylation involved modulation of RNA polymerase II activity. CONCLUSIONS: Our data provide insight into a mechanism by which statins enhance eNOS mRNA stability and increase eNOS protein: statins increase eNOS mRNA polyadenylation through Rho-mediated changes in the actin cytoskeleton. PMID- 17916775 TI - Low c1-inhibitor levels predict early restenosis after eversion carotid endarterectomy. AB - OBJECTIVE: Homozygotes for the normal (A) allele of mannose-binding lectin (MBL2) gene have higher risks to develop an early restenosis after eversion carotid endarterectomy (CEA). Activation of the lectin pathway is regulated by C1 inhibitor (C1-INH). The objective of the present study was to determine the predictive value of C1-INH in restenosis after CEA. METHODS AND RESULTS: C1-INH and MBL-associated serine protease-2 (MASP-2) were determined in samples serially taken from 64 patients with CEA, who were followed-up with carotid duplex scan (CDS) examinations for 14 months. MBL2 genotypes were also determined. Patients with >50% restenosis had lower C1-INH levels at 6 weeks (P=0.0052) and at 4 days (P=0.0277) postsurgery. C1-INH levels at 6 weeks correlated inversely with the CDS values at 14 months (r=-0.3415, P=0.0058), but only in MBL2 A/A homozygotes (r=-0.5044, P=0.0015). Patients with low C1-INH levels (C1-INH <115%) had higher CDS values already at 7 months postsurgery. Patients with MBL2 A/A and low C1-INH levels at 6 weeks postsurgery had 13.97 (95% CI:1.95 to 100.21, P=0.0087) times higher risk to develop an early restenosis. Differences in the MASP-2 concentration were not associated with restenosis. CONCLUSIONS: Determining C1 INH levels at 6 weeks postsurgery-together with genotyping of MBL2-might be a useful marker in the identification of patients with high risk for early carotid restenosis. PMID- 17916776 TI - Increased enzyme activity and beta-adrenergic mediated vasodilation in subjects expressing a single-nucleotide variant of human adenylyl cyclase 6. AB - OBJECTIVE: cAMP is a critical regulator of metabolic and cardiovascular function. However, the role of genetic variability in the regulation of cAMP-mediated effects is unclear. Therefore, we assessed the effect of the expression of a recently identified missense genetic variant of adenylyl cyclase isoform 6 (ADCY6 S674). METHODS AND RESULTS: In rat vascular smooth muscle cells, gene transfer of ADCY6 S674 increased adenylyl cyclase activity and arborization to a greater extent than gene transfer of ADCY6 A674. Similarly, in adherent mononuclear leukocyte cells isolated from ADCY6 S674-expressing human subjects, both adenylyl cyclase activity and adenylyl cyclase-mediated cell retraction were significantly increased. Additionally, in dorsal hand vein LVDT studies, subjects expressing the hyper-functional ADCY6 S674 variant had significantly greater vascular sensitivity to the beta-adrenergic agonist isoproterenol as assessed by both a greater potency and greater maximal effect than subjects expressing the ADCY6 A674 enzyme. CONCLUSIONS: These data indicate that the expression of a novel, relatively common variant of ADCY6 parallels an increase in adenylyl cyclase activity and adenylyl cyclase-mediated function in humans. PMID- 17916778 TI - Preconditioning results in S-nitrosylation of proteins involved in regulation of mitochondrial energetics and calcium transport. AB - Nitric oxide has been shown to be an important signaling messenger in ischemic preconditioning (IPC). Accordingly, we investigated whether protein S nitrosylation occurs in IPC hearts and whether S-nitrosoglutathione (GSNO) elicits similar effects on S-nitrosylation and cardioprotection. Preceding 20 minutes of no-flow ischemia and reperfusion, hearts from C57BL/6J mice were perfused in the Langendorff mode and subjected to the following conditions: (1) control perfusion; (2) IPC; or (3) 0.1 mmol/L GSNO treatment. Compared with control, IPC and GSNO significantly improved postischemic recovery of left ventricular developed pressure and reduced infarct size. IPC and GSNO both significantly increased S-nitrosothiol contents and S-nitrosylation levels of the L-type Ca2+ channel alpha1 subunit in heart membrane fractions. We identified several candidate S-nitrosylated proteins by proteomic analysis following the biotin switch method, including the cardiac sarcoplasmic reticulum Ca2+-ATPase, alpha-ketoglutarate dehydrogenase, and the mitochondrial F1-ATPase alpha1 subunit. The activities of these enzymes were altered in a concentration dependent manner by GSNO treatment. We further developed a 2D DyLight fluorescence difference gel electrophoresis proteomic method that used DyLight fluors and a modified biotin switch method to identify S-nitrosylated proteins. IPC and GSNO produced a similar pattern of S-nitrosylation modification and cardiac protection against ischemia/reperfusion injury, suggesting that protein S nitrosylation may play an important cardioprotective role in heart. PMID- 17916777 TI - Magnetic resonance-based anatomical analysis of scar-related ventricular tachycardia: implications for catheter ablation. AB - In catheter ablation of scar-related monomorphic ventricular tachycardia (VT), substrate voltage mapping is used to electrically define the scar during sinus rhythm. However, the electrically defined scar may not accurately reflect the anatomical scar. Magnetic resonance-based visualization of the scar may elucidate the 3D anatomical correlation between the fine structural details of the scar and scar-related VT circuits. We registered VT activation sequence with the 3D scar anatomy derived from high-resolution contrast-enhanced MRI in a swine model of chronic myocardial infarction using epicardial sock electrodes (n=6, epicardial group), which have direct contact with the myocardium where the electrical signal is recorded. In a separate group of animals (n=5, endocardial group), we also assessed the incidence of endocardial reentry in this model using endocardial basket catheters. Ten to 12 weeks after myocardial infarction, sustained monomorphic VT was reproducibly induced in all animals (n=11). In the epicardial group, 21 VT morphologies were induced, of which 4 (19.0%) showed epicardial reentry. The reentry isthmus was characterized by a relatively small volume of viable myocardium bound by the scar tissue at the infarct border zone or over the infarct. In the endocardial group (n=5), 6 VT morphologies were induced, of which 4 (66.7%) showed endocardial reentry. In conclusion, MRI revealed a scar with spatially complex structures, particularly at the isthmus, with substrate for multiple VT morphologies after a single ischemic episode. Magnetic resonance based visualization of scar morphology would potentially contribute to preprocedural planning for catheter ablation of scar-related, unmappable VT. PMID- 17916779 TI - Targeted deletion of thioredoxin-interacting protein regulates cardiac dysfunction in response to pressure overload. AB - Biomechanical overload induces cardiac hypertrophy and heart failure, and reactive oxygen species (ROS) play a role in both processes. Thioredoxin Interacting Protein (Txnip) is encoded by a mechanically-regulated gene that controls cell growth and apoptosis in part through interaction with the endogenous dithiol antioxidant thioredoxin. Here we show that Txnip is a critical regulator of the cardiac response to pressure overload. We generated inducible cardiomyocyte-specific and systemic Txnip-null mice (Txnip-KO) using Flp/frt and Cre/loxP technologies. Compared with littermate controls, Txnip-KO hearts had attenuated cardiac hypertrophy and preserved left ventricular (LV) contractile reserve through 4 weeks of pressure overload; however, the beneficial effects were not sustained and Txnip deletion ultimately led to maladaptive LV remodeling at 8 weeks of pressure overload. Interestingly, these effects of Txnip deletion on cardiac performance were not accompanied by global changes in thioredoxin activity or ROS; instead, Txnip-KO hearts had a robust increase in myocardial glucose uptake. Thus, deletion of Txnip plays an unanticipated role in myocardial energy homeostasis rather than redox regulation. These results support the emerging concept that the function of Txnip is not as a simple thioredoxin inhibitor but as a metabolic control protein. PMID- 17916781 TI - Introduction. Extremity War Injuries: development of clinical treatment principles. PMID- 17916780 TI - The use of free vascularized fibular grafts in skeletal reconstruction for bone tumors in children. AB - The reconstruction of large skeletal defects in children following resection of a bone tumor presents a unique challenge to the orthopaedic surgeon. Issues in this population that are not present in the adult population include significant remaining growth potential, the desire for biologic preservation of the joint surface, and the need for a long-term viable reconstruction in patients who are anticipated to survive for decades. The use of a free vascularized fibular graft, supplied by the peroneal vessels in intercalary fibular grafts and the anterior tibial vessels in proximal fibular grafts, has been shown to provide biologic reconstruction that successfully addresses these issues in the pediatric population. Specific techniques are applied in the upper and lower extremity to provide long-term excellent functional results. Experience in microvascular surgery and careful postoperative care are required for the success of these procedures. PMID- 17916782 TI - Extremity War Injuries: Development of Clinical Treatment Principles. AB - The AAOS/OTA Extremity War Injuries: Development of Clinical Treatment Principles symposium, held in January 2007, was a follow-up to the first Extremity War Injuries symposium held a year earlier. Discussion focused on four specific areas: prehospital management of extremity wounds, initial debridement, early stabilization, and postoperative wound management during air evacuation. Liberal emergency use of field tourniquets likely is contributing to lower overall mortality and is associated with very low rates of complications. Additional tools for extremity hemorrhage control, such as chitosan-based patches and granular zeolite hemostat, were postulated to be effective. Consensus opinion was that necrotic, devitalized, and contaminated tissue must be removed although objective assessment of completeness of initial debridement is difficult. Definitive open reduction and internal fixation for US and Coalition forces in the theater of operations should be limited to fracture patterns associated with significant perceived risk of delay in treatment. Finally, primary skin closure should be avoided in theater. In addition, because of the time and complications involved in transporting patients to level 4 care facilities, surgeons should consider release of compartments prior to patient transport whenever significant perceived potential for compartment syndrome is present. PMID- 17916783 TI - Chronic whiplash and whiplash-associated disorders: an evidence-based approach. AB - Whiplash is neck pain experienced as a result of a motor vehicle collision or similar trauma. Following a motor vehicle collision, 15% to 40% of patients with acute neck pain develop chronic neck pain. The cervical facet joint is the most common source of chronic neck pain after whiplash injury, followed by disk pain. Some patients experience pain from both structures. Initial management recommendations need not be directed toward an exact structural cause, but treatment includes advising the patient to remain active, prescribing medications when necessary, and providing advice regarding the generally favorable outcome. When neck pain persists, the physician should recommend medial branch blocks of the dorsal rami of the spinal nerves that supply the putative painful facet joint or joints; this is done to determine whether the facet joints are the cause of pain. When significant relief occurs on two occasions, radiofrequency neurotomy typically provides substantial relief for approximately 8 to 12 months and can be repeated indefinitely as needed. Occasionally, long-term treatment with medication may be indicated. Anterior cervical diskectomy and fusion is necessary on rare occasions. PMID- 17916784 TI - Fractures of the greater tuberosity of the humerus. AB - Isolated fractures of the greater tuberosity of the humerus can occur in anterior shoulder dislocations or as the result of an impaction injury against the acromion or superior glenoid. Greater tuberosity fractures may be associated with partial-thickness rotator cuff tears and labral tears, which may be the cause of persistent pain after fracture healing. Nondisplaced and minimally displaced fractures are typically treated successfully nonsurgically. Surgical fixation is recommended for fractures with >5 mm of displacement in the general population or >3 mm of displacement in active patients involved in frequent overhead activity. Open surgical repair is performed with suture or screw fixation. Recently, arthroscopic techniques have produced promising results. Careful follow-up and supervised rehabilitation optimize results after both nonsurgical and surgical treatment. PMID- 17916785 TI - Fixation systems of greater trochanteric osteotomies: biomechanical and clinical outcomes. AB - The development of cerclage systems for fixation of greater trochanteric osteotomies has progressed from monofilament wires to multifilament cables to cable grip and cable plate systems. Cerclage wires and cables have various clinical indications, including fixation for fractures and for trochanteric osteotomy in hip arthroplasty. To achieve stable fixation and eventual union of the trochanteric osteotomy, the implant must counteract the destabilizing forces associated with pull of the peritrochanteric musculature. The material properties of cables and cable grip systems are superior to those of monofilament wires; however, potential complications with the use of cables include debris generation and third-body polyethylene wear. Nevertheless, the cable grip system provides the strongest fixation and results in lower rates of nonunion and trochanteric migration. Cable plate constructs show promise but require further clinical studies to validate their efficacy and safety. PMID- 17916786 TI - Gout affecting the hand and wrist. AB - Tophaceous gout in the hand and wrist often presents de novo as the first sign of the disease process in the elderly. Tophaceous material may present in a liquid, pasty, or chalky/granular state. Treatment may be as simple as aspirating the liquid or squeezing out pasty tophaceous material. Other nonsurgical treatment options include lifestyle and dietary modifications and drug therapy. Surgery is often indicated for the patient with significant tendon and joint compromise as well as skin breakdown and for decompression of compressive peripheral neuropathy. PMID- 17916788 TI - Keynote address: Closing the research-to-practice gap in emergency medicine. AB - Emergency medicine in the United States is facing tremendous challenges due to recent public health emergencies, continuing threats of bioterrorism, and an increasing and unprecedented demand for emergency department services. These challenges include overcrowding; long waiting times; "boarding" of patients; ambulance diversion; a need for better, more reliable tools for triaging patients; and medical errors and other patient safety concerns. These challenges and concerns were brought to the forefront several years ago by the Institute of Medicine in several landmark reports that call for closing the research-to practice gap in emergency medicine. The Agency for Healthcare Research and Quality is funding a number of projects that address many of the concerns raised in the reports, including the use of an advanced access appointment scheduling system to improve access to care; the use of an electronic medical record system to reduce waiting times and errors and improve patient and provider satisfaction; and the refinement of the Emergency Severity Index, a five-level triage scale to get patients to the right resources at the right time. The agency's Healthcare Cost and Utilization Project is gathering data that will allow researchers to examine a broad range of issues affecting the use, quality, and cost of emergency services. Although progress has been made over the past few years in closing the research-to-practice gap in emergency medicine, many challenges remain. The Agency for Healthcare Research and Quality has supported and will continue to support a broad portfolio of research to address the many challenges confronting emergency medicine, including ways to improve emergency care through the application of research findings. PMID- 17916789 TI - Cytochrome P450 1 genes in early deuterostomes (tunicates and sea urchins) and vertebrates (chicken and frog): origin and diversification of the CYP1 gene family. AB - Cytochrome P450 family 1 (CYP1) proteins are important in a large number of toxicological processes. CYP1A and CYP1B genes are well known in mammals, but the evolutionary history of the CYP1 family as a whole is obscure; that history may provide insight into endogenous functions of CYP1 enzymes. Here, we identify CYP1 like genes in early deuterostomes (tunicates and echinoderms), and several new CYP1 genes in vertebrates (chicken, Gallus gallus and frog, Xenopus tropicalis). Profile hidden Markov models (HMMs) generated from vertebrate CYP1A and CYP1B protein sequences were used to identify 5 potential CYP1 homologs in the tunicate Ciona intestinalis genome. The C. intestinalis genes were cloned and sequenced, confirming the predicted sequences. Orthologs of 4 of these genes were found in the Ciona savignyi genome. Bayesian phylogenetic analyses group the tunicate genes in the CYP1 family, provisionally in 2 new subfamilies, CYP1E and CYP1F, which fall in the CYP1A and CYP1B/1C clades. Bayesian and maximum likelihood analyses predict functional divergence between the tunicate and vertebrate CYP1s, and regions within CYP substrate recognition sites were found to differ significantly in position-specific substitution rates between tunicates and vertebrates. Subsequently, 10 CYP1-like genes were found in the echinoderm Strongylocentrotus purpuratus (sea urchin) genome. Several of the tunicate and echinoderm CYP1-like genes are expressed during development. Canonical xenobiotic response elements are present in the upstream genomic sequences of most tunicate and sea urchin CYP1s, and both groups are predicted to possess an aryl hydrocarbon receptor (AHR), suggesting possible regulatory linkage of AHR and these CYPs. The CYP1 family has undergone multiple rounds of gene duplication followed by functional divergence, with at least one gene lost in mammals. This study provides new insight into the origin and evolution of CYP1 genes. PMID- 17916790 TI - Rapid kinetic microassay for catalase activity. AB - Catalase is a commonly assayed enzyme found in many bacteria and eukaryotes. In this report, we examined the applicability of a kinetic microassay to quantify catalase from two different sources. The assay was found to be linear over a wide range (0.1-1.0 units), but was limited at high values (>1 unit) by oxygen evolution. Nonetheless, the microassay allows simultaneous evaluation of many samples (up to 96) in a short time (<5 min) and is thus well-suited to applications, such as high-throughput screening, where many parallel assays are required. PMID- 17916791 TI - A novel approach for ganglioside structural analysis based on electrospray multiple-stage mass spectrometry. AB - A powerful method for detailed structural analysis based on electrospray ionization high-capacity ion-trap multiple-stage mass spectrometry (MS) is for the first time introduced in glycolipidomics. The method was optimized for accurate structural elucidation of human brain gangliosides and specifically applied to normal adult human hippocampus-associated structures. The multiple stage MS experiments reported here allowed for a complete structural characterization of the oligosaccharide moiety of a GM1 ganglioside species. This was achieved by elucidating the sequence and identification of the GM1a structural isomer from the sialic acid attachment site at the neutral oligosaccharide chain. Moreover, the determination of the d18:1/18:0 sphingoid base/fatty acid composition of the ceramide moiety could be confirmed by this method. The novel protocol developed here proves high potential for rapid, reliable, and reproducible investigation of complex lipid-linked carbohydrates such as polysialylated gangliosides or species carrying some other groups that easily cleave off. PMID- 17916792 TI - A two-stage differential hydrogen deuterium exchange method for the rapid characterization of protein/ligand interactions. AB - The peroxisome proliferator-activated receptor is a member of the nuclear receptor superfamily of transcriptional regulators. Regulation of the nuclear receptors occurs through changes to the structure and dynamics of the ligand binding domain. Therefore, the need has arisen for a rapid method capable of detecting changes in the dynamics of nuclear receptors following ligand binding. We recently described how solution-phase amide hydrogen/deuterium exchange (HDX) provides a biophysical technique for probing changes in protein dynamics induced by ligand interaction. Building from this platform, we have optimized the robustness of the differential HDX experiment by minimizing systematic errors, and have increased the efficiency of the chromatographic separation through the use of high-pressure liquid chromatography. Using knowledge gained previously from comprehensive HDX experiments of PPARgamma, a modest throughput method to probe changes in the dynamics of key regions of the receptor was developed. A collection of ten synthetic and endogenous PPARgamma ligands were characterized with this new method requiring approximately 24 h of analysis. This is a dramatic improvement over the 10 d of analysis that would have been required with our previous approach for comprehensive differential HDX analysis. In addition to demonstrating the utility of this approach, the study presented here is the first to measure changes to the dynamics of PPARgamma upon the binding of putative endogenous ligands. PMID- 17916793 TI - Bioinformatic analysis of neural stem cell differentiation. AB - Regulated mRnAs during differentiation of rat neural stem cells were analyzed using the ABi1700 microarray platform. This microarray, while technically advanced, suffers from the difficulty of integrating hybridization results into public databases for systems-level analysis. This is particularly true for the rat array, since many of the probes were designed for transcripts based on predicted human and mouse homologs. using several strategies, we increased the public annotation of the 27,531 probes from 43% to over 65%. To increase the dynamic range of annotation, probes were mapped to numerous public keys from several data sources. consensus annotation from multiple sources was determined for well-scoring alignments, and a confidence-based ranking system established for probes with less agreement across multiple data sources. previous attempts at genomic interpretation using the celera annotation model resulted in poor overlap with expected genomic sequences. since the public keys are more precisely mapped to the genome, we could now analyze the relationships between predicted transcription-factor binding sites and expression clusters. Results collected from a differentiation time course of two neural stem cell clones were clustered using a model-based algorithm. Transcription-factor binding sites were predicted from upstream regions of mapped transcripts using position weight matrices from either JAspAR or TRAnsFAc, and the resulting scores were used to discriminate between observed expression clusters. A classification and regression tree analysis was conducted using cluster numbers as gene identifiers and TFBs scores as predictors, pruning back to obtain a tree with the lowest gene class prediction error rate. Results identify several transcription factors, the presence or absence of which are sufficient to describe clusters of mRnAs changing over time-those that are static, as well as clusters describing cell line differences. public annotation of the AB1700 rat genome array will be valuable for integrating results into future systems-level analyses. PMID- 17916794 TI - A density-based proteomics sample fractionation technology: folate deficiency induced oxidative stress response in liver and brain. AB - Folate deficiency (FD) alters hepatic methionine metabolism and is associated with increased hepatocellular apoptosis. Additionally, mice deprived of folate showed increased oxidative damage in brain tissue, leading to cognitive impairment. Most previous studies have focused independently on either liver, the main tissue of folate storage and metabolism, or brain, where folate regulates neurogenesis and programs cell death. The aim of this study was to apply a powerful, rapid proteomics approach to understand potential subcellular correlations of folate deficiency in both brain and liver of the same rat. This approach combined a new density-based sample fractionation technology (enhanced density gradient extraction = Edge technology) with other conventional proteomics techniques, such as western blot analysis, 2DE, and mass spectrometry. The brain and the liver from individual rats, fed normal or FD diets for 6 wks, were homogenized and then fractionated using the Edge 200 Separation System. Subsequently, all fractions from brain and liver, from control and treated rats, were analyzed by western blot using two markers of oxidative stress: glutathione peroxidase 1 (GPx1) and glucose-regulated protein 75 (GRP75). certain fractions were selected based on western blot analysis and were further analyzed by 2DE. protein spots of interest were identified by MALDI-TOF/TOF. The results demonstrated that edge technology provides a powerful density based separation and enrichment method for rapid screening of potential FD markers and their possible correlations to both liver and brain diseases. PMID- 17916795 TI - A comparison of nLC-ESI-MS/MS and nLC-MALDI-MS/MS for GeLC-based protein identification and iTRAQ-based shotgun quantitative proteomics. AB - The use of nLC-ESI-MS/MS in shotgun proteomics experiments and GeLC-MS/MS analysis is well accepted and routinely available in most proteomics laboratories. However, the same cannot be said for nLC-MALDI MS/MS, which has yet to experience such widespread acceptance, despite the fact that the MALDI technology offers several critical advantages over ESI. As an illustration, in an analysis of moderately complex sample of E. coli proteins, the use MALDI in addition to ESI in GeLC-MS/MS resulted in a 16% average increase in protein identifications, while with more complex samples the number of additional protein identifications increased by an average of 45%. The size of the unique peptides identified by MALDI was, on average, 25% larger than the unique peptides identified by ESI, and they were found to be slightly more hydrophilic. The insensitivity of MALDI to the presence of ionization suppression agents was shown to be a significant advantage, suggesting it be used as a complement to ESI when ion suppression is a possibility. Furthermore, the higher resolution of the TOF/TOF instrument improved the sensitivity, accuracy, and precision of the data over that obtained using only ESI-based iTRAQ experiments using a linear ion trap. Nevertheless, accurate data can be generated with either instrument. These results demonstrate that coupling nanoLC with both ESI and MALDI ionization interfaces improves proteome coverage, reduces the deleterious effects of ionization suppression agents, and improves quantitation, particularly in complex samples. PMID- 17916796 TI - Streamlining plant sample preparation: the use of high-throughput robotics to process echinacea samples for biomarker profiling by MALDI-TOF mass spectrometry. AB - Several species in the genus Echinacea are beneficial herbs popularly used for many ailments. The most popular Echinacea species for cultivation, wild collection, and herbal products include E. purpurea (L.) Moench, E. pallida (Nutt.) Nutt., and E. angustifolia (DC). Product adulteration is a key concern for the natural products industry, where botanical misidentification and introduction of other botanical and nonbotanical contaminants exist throughout the formulation and production process. Therefore, rapid and cost-effective methods that can be used to monitor these materials for complex product purity and consistency are of benefit to consumers and producers. The objective of this continuing research was to develop automated, high-throughput processing methods that, teamed with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, differentiate Echinacea species by their mass profiles. Small molecules, peptide, and proteins from aerial parts (leaf/stem/flowers), seeds, and roots from E. purpurea and E. angustifolia; seeds and roots from E. pallida; and off-the-shelf Echinacea supplements were extracted and analyzed by MS using methods developed on the ProPrep liquid handling system (Genomic Solutions). Analysis of these samples highlighted key MS signal patterns from both small molecules and proteins that characterized the individual Echinacea materials analyzed. Based on analysis of pure Echinacea samples, off the-shelf products containing Echinacea could then be evaluated in a streamlined process. Corresponding analysis of dietary supplements was used to monitor for product composition, including Echinacea species and plant materials used. These results highlight the potential for streamlined, automated approaches for agricultural species differentiation and botanical product evaluation. PMID- 17916797 TI - Antibody microarray analysis of inflammatory mediator release by human leukemia T cells and human non small cell lung cancer cells. AB - Cytokines and chemokines are responsible for regulating inflammation and the immune response. Cytokine and chemokine release is typically measured by quantitative enzyme-linked immunosorbant assay (ELISA) or Western blot analysis. To expedite the analysis of samples for multiple cytokines/chemokines, we have developed slide-based Thermo Scientific ExcelArray Antibody Sandwich Microarrays. Each slide consists of 16 subarrays (wells), each printed with 12 specific antibodies in triplicate and positive and negative control elements. This 16-well format allows for the analysis of 10 test samples using a six-point standard curve. The array architecture is based on the "sandwich" ELISA, in which an analyte protein is sandwiched between an immobilized capture antibody and a biotinylated detection antibody, using streptavidin-linked Thermo Scientific DyLight 649 Dye for quantitation. The observed sensitivity of this assay was <10 pg/mL. In our experiments, the Jurkat cell line was used as a model for human T cell leukemia, and the A549 cell line was used as a model for human non-small cell lung cancer. To evoke a cytokine/chemokine response, cells were stimulated with tumor necrosis factor alpha (TNFalpha), phorbol-12-myristate-13-acetate (PMA, TPA), and phytohemagglutinin (PHA). Cell supernatants derived from both untreated and stimulated cells were analyzed on four different arrays (Inflammation I, Inflammation II, Angiogenesis, and Chemotaxis), enabling the quantitation of 41 unique analytes. Stimulated cells showed an increase in the expression level of many of the test analytes, including IL-8, TNF-alpha, and MIP 1alpha, compared to the non-treated controls. Our experiments clearly demonstrate the utility of antibody microarray analysis of cell-culture supernatants for the profiling of cellular inflammatory mediator release. PMID- 17916798 TI - Analysis of glycoproteins in human serum by means of glycospecific magnetic bead separation and LC-MALDI-TOF/TOF analysis with automated glycopeptide detection. AB - Comprehensive proteomic analyses require efficient and selective pre fractionation to facilitate analysis of post-translationally modified peptides and proteins, and automated analysis workflows enabling the detection, identification, and structural characterization of the corresponding peptide modifications. Human serum contains a high number of glycoproteins, comprising several orders of magnitude in concentration. Thereby, isolation and subsequent identification of low-abundant glycoproteins from serum is a challenging task. selective capturing of glycopeptides and -proteins was attained by means of magnetic particles specifically functionalized with lectins or boronic acids that bind to various structural motifs. Human serum was incubated with differentially functionalized magnetic micro-particles (lectins or boronic acids), and isolated proteins were digested with trypsin. Subsequently, the resulting complex mixture of peptides and glycopeptides was subjected to LC-MALDI analysis and database searching. In parallel, a second magnetic bead capturing was performed on the peptide level to separate and analyze by LC-MALDI intact glycopeptides, both peptide sequence and glycan structure. Detection of glycopeptides was achieved by means of a software algorithm that allows extraction and characterization of potential glycopeptide candidates from large LC-MALDI-MS/MS data sets, based on N glycopeptide-specific fragmentation patterns and characteristic fragment mass peaks, respectively. By means of fast and simple glycospecific capturing applied in conjunction with extensive LC-MALDI-MS/MS analysis and novel data analysis tools, a high number of low-abundant proteins were identified, comprising known or predicted glycosylation sites. According to the specific binding preferences of the different types of beads, complementary results were obtained from the experiments using either magnetic ConA-, LCA-, WGA-, and boronic acid beads, respectively. PMID- 17916799 TI - Measuring the binding stoichiometry of HIV-1 Gag to very-low-density oligonucleotide surfaces using surface plasmon resonance spectroscopy. AB - The interaction of the HIV Gag polyprotein with nucleic acid is a critical step in the assembly of viral particles. The Gag polyprotein is composed of the matrix (MA), capsid (CA), and nucleocapsid (NC) domains. The NC domain is required for nucleic acid interactions, and the CA domain is required for Gag-Gag interactions. Previously, we have investigated the binding of the NC protein to d(TG)(n) oligonucleotides using surface plasmon resonance (SPR) spectroscopy. We found a single NC protein is able to bind to more than one immobilized oligonucleotide, provided that the oligonucleotides are close enough together. As NC is believed to be the nucleic acid binding domain of Gag, we might expect Gag to show the same complex behavior. We wished to analyze the stoichiometry of Gag binding to oligonucleotides without this complication due to tertiary complex formation. We have therefore analyzed Gag binding to extremely low oligonucleotide density on SPR chips. Such low densities of oligonucleotides are difficult to accurately quantitate. We have determined by Fourier transform ion cyclotron (FTICR) mass spectrometry that four molecules of NC bind to d(TG)(10) (a 20-base oligonucleotide). We developed a method of calibrating low-density surfaces using NC calibration injections. Knowing the maximal response and the stoichiometry of binding, we can precisely determine the amount of oligonucleotide immobilized at these very-low-density surfaces (<1 Response Unit). Using this approach, we have measured the binding of Gag to d(TG)(10). Gag binds to a 20-mer with a stoichiometry of greater than 4. This suggests that once Gag is bound to the immobilized oligonucleotide, additional Gag molecules can bind to this complex. PMID- 17916800 TI - Toll-like receptors on human mesenchymal stem cells drive their migration and immunomodulating responses. AB - Adult human bone marrow-derived mesenchymal stem cells (hMSCs) are under study as therapeutic delivery agents that assist in the repair of damaged tissues. To achieve the desired clinical outcomes for this strategy requires a better understanding of the mechanisms that drive the recruitment, migration, and engraftment of hMSCs to the targeted tissues. It is known that hMSCs are recruited to sites of stress or inflammation to fulfill their repair function. It is recognized that toll-like receptors (TLRs) mediate stress responses of other bone marrow-derived cells. This study explored the role of TLRs in mediating stress responses of hMSCs. Accordingly, the presence of TLRs in hMSCs was initially established by reverse transcription-polymerase chain reaction assays. Flow cytometry and fluorescence immunocytochemical analyses confirmed these findings. The stimulation of hMSCs with TLR agonists led to the activation of downstream signaling pathways, including nuclear factor kappaB, AKT, and MAPK. Consequently, activation of these pathways triggered the induction and secretion of cytokines, chemokines, and related TLR gene products as established from cDNA array, immunoassay, and cytokine antibody array analyses. Interestingly, the unique patterns of affected genes, cytokines, and chemokines measured identify these receptors as critical players in the clinically established immunomodulation observed for hMSCs. Lastly, hMSC migration was promoted by TLR ligand exposure as demonstrated by transwell migration assays. Conversely, disruption of TLRs by neutralizing TLR antibodies compromised hMSC migration. This study defines a novel TLR-driven stress and immune modulating response for hMSCs that is critical to consider in the design of stem cell-based therapies. PMID- 17916801 TI - Human multipotent stromal cells undergo sharp transition from division to development in culture. AB - Human mesenchymal stem cells, or multipotent stromal cells (MSCs), are of interest for clinical therapy, in part because of their capacity for proliferation and differentiation. However, results from clinical trials and in vitro models have been variable, possibly because of MSC heterogeneity and a lack of standardization between MSC in vitro expansion protocols. Here we defined changes in MSCs during expansion in vitro. In low-density cultures, MSCs expand through distinct lag, exponential growth, and stationary phases. We assayed cultures of passage 2 human MSCs from three donors at low density (50 cells per cm(2)) at approximately 5% confluence on day 2 and after the cultures had expanded to approximately 70% confluence on day 7. On day 2, genes involved in cell division were upregulated. On day 7, genes for cell development were upregulated. The variations among three donors were less than the variation within the expansion of MSCs from a single donor. The microarray data for selected genes were confirmed by real-time polymerase chain reaction, enzyme linked immunosorbent assay, and FACScan analysis. Approximately 50% of cells at day 2 were in S-phase compared with 10% at day 7. The results demonstrated major differences in early and late stage cultures of MSCs that should be considered in using the cells in experiments and clinical applications. PMID- 17916802 TI - Myocardin a enhances telomerase activities in adipose tissue mesenchymal cells and embryonic stem cells undergoing cardiovascular myogenic differentiation. AB - Acting as a reverse transcriptase that maintains nuclear telomere length and replication potential, telomerase usually decreases in expression and activities when mammalian stem cells undergo terminal differentiation. This study identified, in adult adipose tissue, a subpopulation of mesenchymal stem cells (MSCs) that coexpresses telomerase and myocardin A, a key regulator of cardiovascular myogenic development. The telomerase/myocardin A-positive MSCs differentiated into cardiovascular myogenic cells while retaining expression and activation of the telomerase catalytic unit, telomerase reverse transcriptase (TERT), at a level comparable to that of ESCs. Both myocardin A and TERT could be coimmunoprecipitated from the developing MSCs and ESC-derived EBs with either anti-TERT or anti-myocardin A antibodies, suggesting the formation of TERT myocardin A complexes in the MSCs and EBs. The proteins pulled down with anti myocardin antibodies showed almost the same levels of telomerase activities as those precipitated with anti-TERT antibodies. Overexpression of myocardin A by cDNA transfection significantly increased telomerase activities and promoted telomere synthesis by MSCs. The data from this study indicate a potentially novel function of myocardin A in maintaining the myogenic stemness in developing MSCs and EBs by enhancing telomerase activation and promoting myogenic gene expression. PMID- 17916804 TI - MicroRNA-134 modulates the differentiation of mouse embryonic stem cells, where it causes post-transcriptional attenuation of Nanog and LRH1. AB - Hundreds of microRNAs (miRNAs) are expressed in mammalian cells, where they aid in modulating gene expression by mediating mRNA transcript cleavage and/or regulation of translation rate. Functional studies to date have demonstrated that several of these miRNAs are important during development. However, the role of miRNAs in the regulation of stem cell growth and differentiation is not well understood. We show herein that microRNA (miR)-134 levels are maximally elevated at day 4 after retinoic acid-induced differentiation or day 2 after N2B27-induced differentiation of mouse embryonic stem cells (mESCs), but this change is not observed during embryoid body differentiation. The elevation of miR-134 levels alone in mESCs enhances differentiation toward ectodermal lineages, an effect that is blocked by a miR-134 antagonist. The promotion of mESC differentiation by miR-134 is due, in part, to its direct translational attenuation of Nanog and LRH1, both of which are known positive regulators of Oct4/POU5F1 and mESC growth. Together, the data demonstrate that miR-134 alone can enhance the differentiation of mESCs to ectodermal lineages and establish a functional role for miR-134 in modulating mESC differentiation through its potential to target and regulate multiple mRNAs. PMID- 17916803 TI - SFRP2 regulates cardiomyogenic differentiation by inhibiting a positive transcriptional autofeedback loop of Wnt3a. AB - Wnts comprise a family of 20 lipid-modified glycoproteins in mammals and play critical roles during embryological development and organogenesis of several organ systems, including the heart. They are required for mesoderm formation and have been implicated in promoting cardiomyogenic differentiation of mammalian embryonic stem cells, but the underlying mechanisms regulating Wnt signaling during cardiomyogenesis remain poorly understood. In this report, we show that in a pluripotent mouse embryonal carcinoma stem cell line, SFRP2 inhibits cardiomyogenic differentiation by regulating Wnt3a transcription. SFRP2 inhibited early stages of cardiomyogenesis, preventing mesoderm specification and maintaining the cells in the undifferentiated state. Using a gain- and loss-of function approach, we demonstrate that although addition of recombinant SFRP2 decreased Wnt3a transcription and cardiomyogenic differentiation, silencing of Sfrp2 led to enhanced Wnt3a transcription, mesoderm formation, and increased cardiomyogenesis. We show that the inhibitory effects of SFRP2 on Wnt transcription are secondary to interruption of a positive feedback effect of Wnt3a on its own transcription. Wnt3a increased its own transcription via the canonical pathway and TCF4 family of transcription factors, and the inhibitory effects of SFRP2 on Wnt3a transcription were associated with disruption of downstream canonical Wnt signaling. The inhibitory effects of Sfrp2 on Wnt3a expression identify Sfrp2 as a "checkpoint gene," which exerts its control on cardiomyogenesis through regulation of Wnt3a transcription. PMID- 17916805 TI - Impairment of apoptotic cell engulfment by pyocyanin, a toxic metabolite of Pseudomonas aeruginosa. AB - RATIONALE: Cystic fibrosis lung disease is characterized by accumulation of apoptotic neutrophils, indicating impaired clearance of dying cells. Pseudomonas aeruginosa, the principal microbial pathogen in cystic fibrosis, manipulates apoptosis induction via production of toxic metabolites. Whether these metabolites, particularly pyocyanin, can also modulate apoptotic cell engulfment is unknown. OBJECTIVES: To assess the effects of pyocyanin on apoptotic cell engulfment by macrophages in vitro and in vivo and to investigate potential mechanisms of the observed effects. METHODS: Human monocyte-derived macrophages were treated with pyocyanin before challenge with apoptotic neutrophils, apoptotic Jurkat cells, or latex beads, and phagocytosis was assessed by light microscopy and flow cytometry. Effects of pyocyanin production on apoptotic cell clearance in vivo were assessed in a murine model, comparing infection by wild type or pyocyanin-deficient P. aeruginosa. Oxidant production was investigated using fluorescent probes and pharmacologic inhibition and Rho GTPase signaling by immunoblotting and inhibitor studies. MEASUREMENTS AND MAIN RESULTS: Pyocyanin treatment impaired macrophage engulfment of apoptotic cells in vitro, without inducing significant macrophage apoptosis, whereas latex bead uptake was preserved. Macrophage ingestion of apoptotic cells was reduced and late apoptotic/necrotic cells were increased in mice infected with pyocyanin-producing P. aeruginosa compared with the pyocyanin-deficient strain. Inhibition of apoptotic cell uptake involved intracellular generation of reactive oxygen species (ROS) and effects on Rho GTPase signaling. Antioxidants or blockade of Rho signaling substantially restored apoptotic cell engulfment. CONCLUSIONS: These studies demonstrate that P. aeruginosa can manipulate the inflammatory microenvironment through inhibition of apoptotic cell engulfment, and suggest potential strategies to limit pulmonary inflammation in cystic fibrosis. PMID- 17916806 TI - The prevention of chronic obstructive pulmonary disease exacerbations by salmeterol/fluticasone propionate or tiotropium bromide. AB - RATIONALE: Exacerbations are key drivers of morbidity and mortality in chronic obstructive pulmonary disease (COPD). OBJECTIVES: We compared the relative efficacy of the long-acting inhaled bronchodilator/antiinflammatory combination (salmeterol/fluticasone propionate) 50/500 microg twice daily and the long-acting bronchodilator (tiotropium) 18 microg once daily in preventing exacerbations and related outcomes in severe and very severe COPD. METHODS: A total of 1,323 patients (mean age, 64 yr, post-bronchodilator FEV1, 39% predicted) were randomized in this 2-year, double-blind, double-dummy parallel study. MEASUREMENTS AND MAIN RESULTS: Primary endpoint was health care utilization exacerbation rate. Other endpoints included health status measured by St. George's Respiratory Questionnaire (SGRQ), mortality, adverse events, and study withdrawal. Probability of withdrawing from the study was 29% greater with tiotropium than salmeterol/fluticasone propionate (P = 0.005). The modeled annual exacerbation rate was 1.28 in the salmeterol/fluticasone propionate group and 1.32 in the tiotropium group (rate ratio, 0.967; 95% confidence interval [CI], 0.836-1.119]; P = 0.656). The SGRQ total score was statistically significantly lower at 2 years on salmeterol/fluticasone propionate versus tiotropium (difference 2.1 units; 95% CI, 0.1-4.0; P = 0.038). Mortality was significantly lower in the salmeterol/fluticasone propionate group; 21 (3%) of patients in this group died compared with 38 (6%) in the tiotropium group (P = 0.032). More pneumonias were reported in the salmeterol/fluticasone propionate group relative to tiotropium (P = 0.008). CONCLUSIONS: We found no difference in exacerbation rate between salmeterol/fluticasone propionate and tiotropium. More patients failed to complete the study while receiving tiotropium. A small statistically significant beneficial effect was found on health status, with an unexpected finding of lower deaths in salmeterol/fluticasone propionate-treated patients. Clinical trial registered with www.clinicaltrials.gov (NCT 00361959). PMID- 17916807 TI - Variable prostaglandin E2 resistance in fibroblasts from patients with usual interstitial pneumonia. AB - RATIONALE: Prostaglandin (PG) E2, a cyclooxygenase-derived lipid mediator, is a potent down-regulator of fibroblast activation in normal lung fibroblasts. Although fibroblasts from patients with idiopathic pulmonary fibrosis are known to exhibit a defect in PGE2 synthesis, there is little information about their responsiveness to this lipid mediator. OBJECTIVES: To compare responses to PGE2 in normal, usual interstitial pneumonia (UIP), and other diffuse parenchymal lung disease (DPLD) fibroblasts. METHODS: Fibroblasts were grown in vitro from well characterized control (n = 7), UIP (n = 17), or other DPLD (n = 13) lung tissue. The effects of PGE2 on fibroblast proliferation and collagen expression were determined. MEASUREMENTS AND MAIN RESULTS: Only 3 of 12 UIP fibroblast lines exhibited PGE2-mediated inhibition of both collagen synthesis and cell proliferation, as opposed to 6 of 6 nonfibrotic control cell lines. The degree of PGE2 resistance in DPLD fibroblasts was quite variable, with UIP cells exhibiting the greatest degree of resistance to PGE2, whereas other DPLD fibroblasts manifested a degree of resistance intermediate to control and UIP. The resistance to suppression of collagen expression correlated with worse lung function. Molecular mechanisms for resistance included altered E prostanoid receptor profiles and diminished expression of the downstream kinase, protein kinase A. CONCLUSIONS: The recognition that UIP fibroblasts manifest variable refractoriness to PGE2 suppression sheds new light on the activation phenotype of these cells and on the pathogenesis of fibrotic lung disease. PMID- 17916808 TI - Inhibition of integrin alpha(v)beta6, an activator of latent transforming growth factor-beta, prevents radiation-induced lung fibrosis. AB - RATIONALE: In experimental models, lung fibrosis is dependent on transforming growth factor (TGF)-beta signaling. TGF-beta is secreted in a latent complex with its propeptide, and TGF-beta activators release TGF-beta from this complex. Because the integrin alpha(v)beta6 is a major TGF-beta activator in the lung, inhibition of alpha(v)beta6-mediated TGF-beta activation is a logical strategy to treat lung fibrosis. OBJECTIVES: To determine, by genetic and pharmacologic approaches, whether murine radiation-induced lung fibrosis is dependent on alpha(v)beta6. METHODS: Wild-type mice, alpha(v)beta6-deficient (Itgb6-/-) mice, and mice heterozygous for a Tgfb1 mutation that eliminates integrin-mediated activation (Tgfb1(+/RGE)) were exposed to 14 Gy thoracic radiation. Some mice were treated with an anti-alpha(v)beta6 monoclonal antibody or a soluble TGF-beta receptor fusion protein. Alpha(v)beta6 expression was determined by immunohistochemistry. Fibrosis, inflammation, and gene expression patterns were assessed 20-32 weeks postirradiation. MEASUREMENTS AND MAIN RESULTS: Beta6 integrin expression increased within the alveolar epithelium 18 weeks postirradiation, just before onset of fibrosis. Itgb6-/- mice were completely protected from fibrosis, but not from late radiation-induced mortality. Anti alpha(v)beta6 therapy (1-10 mg/kg/wk) prevented fibrosis, but only higher doses (6-10 mg/kg/wk) caused lung inflammation similar to that in Itgb6-/- mice. Tgfb1 haploinsufficient mice were also protected from fibrosis. CONCLUSIONS: Alpha(v)beta6-mediated TGF-beta activation is required for radiation-induced lung fibrosis. Together with previous data, our results demonstrate a robust requirement for alpha(v)beta6 in distinct fibrosis models. Inhibition of alphavbeta6-mediated TGF-beta activation is a promising new approach for antifibrosis therapy. PMID- 17916809 TI - Partial inhibition of integrin alpha(v)beta6 prevents pulmonary fibrosis without exacerbating inflammation. AB - RATIONALE: Transforming growth factor (TGF)-beta has a central role in driving many of the pathological processes that characterize pulmonary fibrosis. Inhibition of the integrin alpha(v)beta6, a key activator of TGF-beta in lung, is an attractive therapeutic strategy, as it may be possible to inhibit TGF-beta at sites of alpha(v)beta6 up-regulation without affecting other homeostatic roles of TGF-beta. OBJECTIVES: To analyze the expression of alpha(v)beta6 in human pulmonary fibrosis, and to functionally test the efficacy of therapeutic inhibition of alpha(v)beta6-mediated TGF-beta activation in murine bleomycin induced pulmonary fibrosis. METHODS: Lung biopsies from patients with a diagnosis of systemic sclerosis or idiopathic pulmonary fibrosis were stained for alpha(v)beta6 expression. A range of concentrations of a monoclonal antibody that blocks alpha(v)beta6-mediated TGF-beta activation was evaluated in murine bleomycin-induced lung fibrosis. MEASUREMENTS AND MAIN RESULTS: Alpha(v)beta6 is overexpressed in human lung fibrosis within pneumocytes lining the alveolar ducts and alveoli. In the bleomycin model, alpha(v)beta6 antibody was effective in blocking pulmonary fibrosis. At high doses, there was increased expression of markers of inflammation and macrophage activation, consistent with the effects of TGF-beta inhibition in the lung. Low doses of antibody attenuated collagen expression without increasing alveolar inflammatory cell populations or macrophage activation markers. CONCLUSIONS: Partial inhibition of TGF-beta using alpha(v)beta6 integrin antibodies is effective in blocking murine pulmonary fibrosis without exacerbating inflammation. In addition, the elevated expression of alpha(v)beta6, an activator of the fibrogenic cytokine, TGF-beta, in human pulmonary fibrosis suggests that alpha(v)beta6 monoclonal antibodies could represent a promising new therapeutic strategy for treating pulmonary fibrosis. PMID- 17916810 TI - Boxing and the risk of chronic brain injury. PMID- 17916811 TI - Amateur boxing and risk of chronic traumatic brain injury: systematic review of observational studies. AB - OBJECTIVE: To evaluate the risk of chronic traumatic brain injury from amateur boxing. SETTING: Secondary research performed by combination of sport physicians and clinical academics. DESIGN, DATA SOURCES, AND METHODS: Systematic review of observational studies in which chronic traumatic brain injury was defined as any abnormality on clinical neurological examination, psychometric testing, neuroimaging studies, and electroencephalography. Studies were identified through database (1950 to date) and bibliographic searches without language restrictions. Two reviewers extracted study characteristics, quality, and data, with adherence to a protocol developed from a widely recommended method for systematic review of observational studies (MOOSE). RESULTS: 36 papers had relevant extractable data (from a detailed evaluation of 93 studies of 943 identified from the initial search). Quality of evidence was generally poor. The best quality studies were those with a cohort design and those that used psychometric tests. These yielded the most negative results: only four of 17 (24%) better quality studies found any indication of chronic traumatic brain injury in a minority of boxers studied. CONCLUSION: There is no strong evidence to associate chronic traumatic brain injury with amateur boxing. PMID- 17916812 TI - Paramedic practitioners and emergency admissions. PMID- 17916813 TI - Effectiveness of paramedic practitioners in attending 999 calls from elderly people in the community: cluster randomised controlled trial. AB - OBJECTIVE: To evaluate the benefits of paramedic practitioners assessing and, when possible, treating older people in the community after minor injury or illness. Paramedic practitioners have been trained with extended skills to assess, treat, and discharge older patients with minor acute conditions in the community. DESIGN: Cluster randomised controlled trial involving 56 clusters. Weeks were randomised to the paramedic practitioner service being active (intervention) or inactive (control) when the standard 999 service was available. SETTING: A large urban area in England. PARTICIPANTS: 3018 patients aged over 60 who called the emergency services (n=1549 intervention, n=1469 control). MAIN OUTCOME MEASURES: Emergency department attendance or hospital admission between 0 and 28 days; interval from time of call to time of discharge; patients' satisfaction with the service received. RESULTS: Overall, patients in the intervention group were less likely to attend an emergency department (relative risk 0.72, 95% confidence interval 0.68 to 0.75) or require hospital admission within 28 days (0.87, 0.81 to 0.94) and experienced a shorter total episode time (235 v 278 minutes, 95% confidence interval for difference -60 minutes to -25 minutes). Patients in the intervention group were more likely to report being highly satisfied with their healthcare episode (relative risk 1.16, 1.09 to 1.23). There was no significant difference in 28 day mortality (0.87, 0.63 to 1.21). CONCLUSIONS: Paramedics with extended skills can provide a clinically effective alternative to standard ambulance transfer and treatment in an emergency department for elderly patients with acute minor conditions. TRIAL REGISTRATION: ISRCTN27796329 [controlled-trials.com]. PMID- 17916814 TI - Encouraging children and adolescents to be more active. PMID- 17916815 TI - Occlusion therapy for amblyopia. PMID- 17916816 TI - Reform of the coroner system and death certification. PMID- 17916817 TI - Diagnosing left ventricular hypertrophy in arterial hypertension. PMID- 17916818 TI - Direct to consumer advertisement: The world of the market place. PMID- 17916819 TI - Familial hypercholesterolaemia: Child-parent screening may have adverse psychological effects. PMID- 17916820 TI - Familial hypercholesterolaemia: Cascade testing is tried and tested and cost effective. PMID- 17916821 TI - Bare below the elbows: A cheap soundbite. PMID- 17916822 TI - Medical migration: The real elephant in the room. PMID- 17916823 TI - Medical migration: Full house. PMID- 17916824 TI - Future of US children's health insurance still uncertain. PMID- 17916825 TI - Africans die in pain because of fears of opiate addiction. PMID- 17916826 TI - UK does well on giving information to patients but poorly on access to new treatments. PMID- 17916827 TI - Netherlands considers introducing preconception care. PMID- 17916828 TI - African-American leaders call for concerted action on AIDS in US. PMID- 17916829 TI - Italian court upholds couple's demand for preimplantation genetic diagnosis. PMID- 17916830 TI - Patients win right to have their advance decisions honoured. PMID- 17916834 TI - Treat elderly people with dignity, trusts warned. PMID- 17916835 TI - Doctors who help give lethal injections should be punished, says Amnesty. PMID- 17916839 TI - FDA is failing to oversee human clinical trials, report says. PMID- 17916840 TI - Doctors disagree about directive on European clinical trials. PMID- 17916841 TI - Australian efforts to tackle abuse of Aboriginal children raise alarm. PMID- 17916850 TI - Industry funded patient information and the slippery slope to New Zealand. PMID- 17916851 TI - Should general practitioners resume 24 hour responsibility for their patients? Yes. PMID- 17916852 TI - Should general practitioners resume 24 hour responsibility for their patients? No. PMID- 17916853 TI - An age old problem. PMID- 17916854 TI - Adding fluoride to water supplies. PMID- 17916855 TI - Colorectal cancer. PMID- 17916856 TI - Assessment, investigation, and early management of head injury: summary of NICE guidance. PMID- 17916857 TI - Uterine artery embolisation to treat symptomatic uterine fibroids. PMID- 17916863 TI - Peer support in an occupational setting preventing LBP-related sick leave. AB - BACKGROUND: Low back pain (LBP) is among the most frequent causes of sickness absence in Norway, and it is thought that it could be reduced by 30-50% if present day knowledge was implemented in the workplace. Evidence-based interventions in occupational settings to prevent sickness absence are still lacking. AIM: To evaluate whether peer support would be able to modify general beliefs about LBP, pain experiences, health care utilization and sickness absence due to back pain. METHODS: In addition to a media campaign in two Norwegian counties in 2002-05, aiming at improving beliefs about LBP in the general public, the 'Active Back' project trained a peer adviser in six participating workplaces. The task of this peer adviser was to provide information aimed at reducing fear of the pain, supportive advice and arrange for modifications of workloads, etc., for a limited period of time. RESULTS: The prevalence of back pain remained constant throughout the study period, but self-reported intensity of LBP decreased at the end. There was a small decline in use of health care professionals and significant improvements in beliefs, in line with the messages of the campaign. Total sickness absence decreased by 27% and the LBP-related sickness absence by 49%. CONCLUSION: The combination of peer support and modified workload seemed to have additional effects to the general media campaign, and resulted in decline in sickness absence and improvements in beliefs about back pain. PMID- 17916864 TI - A case of hepatomegaly. PMID- 17916865 TI - Once weekly thyroxine treatment as a strategy to treat non-compliance. AB - Hypothyroidism is a common disorder, which is mainly treated in primary rather than secondary care. Once daily thyroxine replacement restores euthyroidism in most patients; some patients, however, remain hypothyroid despite adequate thyroxine replacement. Non-compliance is the most common cause of lack of response to thyroxine treatment. We describe two cases of primary hypothyroidism in which daily thyroxine treatment did not restore biochemical euthyroidism but once weekly thyroxine treatment was successful. In addition we review the evidence and discuss the differential diagnosis of lack of response to thyroxine treatment. Once weekly thyroxine treatment can be a safe, well-tolerated, and effective therapy for patients with non-compliance. PMID- 17916867 TI - Herbal medicine: buy one, get two free. PMID- 17916866 TI - One-year-old male with accelerated growth and development. AB - A 1-year-old male child with isosexual central (gonadotropin-dependent) precocious puberty caused by hypothalamic hamartoma is reported. Details of the diagnosis based solely on neuromaging characteristics, and satisfactory results of medical treatment with gonadotropin releasing hormone agonist analogues, are highlighted. PMID- 17916868 TI - A proposal justifying an alternative referral practice from primary care for three common hand surgery diagnoses. PMID- 17916869 TI - Sarcoid heart disease. AB - To this day the aetiology of sarcoidosis continues to elude definition. Partially as a consequence of this, little in the way of new therapies has evolved. The enigma of this condition is that, unusually for a disease with the potential for devastating consequences, many patients show spontaneous resolution and recover. Cardiac involvement can affect individuals of any age, gender or race and has a predilection for the conduction system of the heart. Heart involvement can also cause a dilated cardiomyopathy with consequent progressive heart failure. The most common presentation of this systemic disease is with pulmonary infiltration, but many cases will be asymptomatic and are detected on routine chest radiography revealing lymphadenopathy. Current advances lie in the newer methods of imaging and diagnosing this unusual heart disease. This review describes the pathology and diagnosis of this condition and the newer imaging techniques that have developed for determining cardiac involvement. PMID- 17916870 TI - Testis cancer. AB - Testis cancer is an increasing problem, especially in northern European male populations. However, survival has improved dramatically over one generation. Environmental factors may have a role in the aetiology with high oestrogen concentrations implicated. Testis cancer is subdivided between seminoma and non seminoma. At presentation, a testicular lump is the most common finding and radical inguinal orchidectomy is recommended for most. Further multidisciplinary management is determined by histological subtype and stage and involves chemotherapy, radiotherapy and surgery, with many patients only undergoing surveillance. There is increasing emphasis on reducing toxicity of treatments in long term survivors. Treatment refractory testis cancer remains a significant challenge. PMID- 17916871 TI - A systematic review of randomised clinical trials of individualised herbal medicine in any indication. AB - AIM: To summarise and critically evaluate the evidence from randomised clinical trials for the effectiveness of individualised herbal medicine in any indication. METHODS: Search of electronic databases and approaches to experts in the field to identify randomised, controlled clinical trials of individualised herbal medicine in any indication. Independent data extraction and assessment of methodological quality by two authors and best evidence synthesis. RESULTS: Three randomised clinical trials of individualised herbal medicine were identified. Statistically non-significant trends favouring active over placebo treatment in osteoarthritis of the knee probably result from large baseline differences and regression to the mean. Individualised treatment was superior to placebo in four of five outcome measures in the treatment of irritable bowel syndrome, but was inferior to standardised herbal treatment in all outcomes. Individualised herbal treatment was no better than placebo in the prevention of chemotherapy-induced toxicity. CONCLUSIONS: There is a sparsity of evidence regarding the effectiveness of individualised herbal medicine and no convincing evidence to support the use of individualised herbal medicine in any indication. PMID- 17916872 TI - Current management of male-to-female gender identity disorder in the UK. AB - Gender identity disorder (GID), or transsexualism as it is more commonly known, is a highly complex clinical entity. Although the exact aetiology of GID is unknown, several environmental, genetic and anatomical theories have been described. The diagnosis of GID can be a difficult process but is established currently using standards of care as defined by the Harry Benjamin International Gender Dysphoria Association. Patients go through extensive psychiatric assessment, including the Real Life Experience, which entails living in the desired gender role 24 h a day for a minimum period of 12 months. The majority of GID patients will eventually go on to have gender realignment surgery, which includes feminising genitoplasty. The clinical features, diagnostic approach and management of male-to-female GID in the UK are reviewed, including the behavioural, psychological and surgical aspects. PMID- 17916873 TI - "Please, I want to go home": ethical issues raised when considering choice of place of care in palliative care. AB - Many palliative care patients would prefer to receive care, and to die, at home. Despite this many die in institutions. In response to this, politicians and charities have adopted policies aimed at increasing the opportunities for care and death at home. The need to discuss plans for discharge with most inpatients reinforces expectations of a choice of place of care. However, many palliative patients do not have a choice of care at home. This article will explore the circumstances in which patients are unable to choose home care and consider changes in clinical practice that can help to maximise choice. We shall argue that there is a distinction between the preferences of patients and the choices actually available to them. In attempting to make this distinction we advocate consideration of the balance between the ethical principles relevant to each case. PMID- 17916875 TI - A description of the methods used to obtain information on ancient disease and medicine and of how the evidence has survived. AB - This paper summarises the common modalities that are available for researching ancient medicine and disease as well as explaining how some of these sources have survived to modern day. These are explained under the three broad headings of palaeopathology, artefacts, and texts. The descriptions use a variety of examples from ancient societies including in the Bronze Age, Babylonia and Assyria, ancient Egypt, ancient Greece, and ancient Rome to help explain these modalities. In addition, a review of the advantages and disadvantages of using these tools is included to help current and future historians in stimulating future research in this fascinating area. PMID- 17916874 TI - Recent advances and controversies in adult cardiopulmonary resuscitation. AB - Since its introduction more than four and half decades ago, the science of cardiopulmonary resuscitation has been enriched with a significant amount of scientific evidence. This in turn has led to the birth of new evidence based guidelines for resuscitation published by the European Resuscitation Council and the American Heart Association in late 2005. This article aims to review the recent advances and controversies in the science of resuscitation. PMID- 17916876 TI - Effect of an acute medical admission unit on key quality indicators assessed by funnel plots. AB - OBJECTIVE: To determine the effect of the introduction of an acute medical admissions unit (AMAU) on key quality efficiency and outcome indicator comparisons between medical teams as assessed by funnel plots. METHODS: A retrospective analysis was performed of data relating to emergency medical patients admitted to St James' Hospital, Dublin between 1 January 2002 and 31 December 2004, using data on discharges from hospital recorded in the hospital in patient enquiry system. The base year was 2002 during which patients were admitted to a variety of wards under the care of a named consultant physician. In 2003, two centrally located wards were reconfigured to function as an AMAU, and all emergency patients were admitted directly to this unit. The quality indicators examined between teams were length of stay (LOS) <30 days, LOS >30 days, and readmission rates. RESULTS: The impact of the AMAU reduced overall hospital LOS from 7 days in 2002 to 5 days in 2003/04 (p<0.0001). There was no change in readmission rates between teams over the 3 year period, with all teams displaying expected variability within control (95%) limits. Overall, the performance in LOS, both short term and long term, was significantly improved (p<0.0001), and was less varied between medical teams between 2002 and 2003/04. CONCLUSIONS: Introduction of the AMAU improved performance among medical teams in LOS, both short term and long term, with no change in readmissions. Funnel plots are a powerful graphical technique for presenting quality performance indicator variation between teams over time. PMID- 17916877 TI - Intravenous administration of metoclopramide by 2 min bolus vs 15 min infusion: does it affect the improvement of headache while reducing the side effects? AB - OBJECTIVE: To determine the therapeutic effect (alleviation of vascular type headache) and side effects of a slow intravenous metoclopramide infusion over 15 min compared with those effects of a bolus intravenous metoclopramide infusion over 2 min in the treatment of patients with recent onset vascular type headache. MATERIAL AND METHODS: All adults treated with metoclopramide for vascular type headache were eligible for entry into this clinical randomised double blinded trial. This study compared the effects of two different rates of intravenous infusion of metoclopramide over a period of 13 months at a university hospital emergency department. During the trial, side effects and headache scores were recorded at baseline (0 min), and then at 5, 15, 30 and 60 min. Repeated measures analysis of variance was used to compare the medication's efficacy and side effects. RESULTS: A total of 120 patients presenting to the emergency department met the inclusion criteria. Of these, 62 patients (51.7%) were given 10 mg metoclopramide as a slow intravenous infusion over 15 min (SIG group) and 58 patients (48.3%) were given 10 mg metoclopramide intravenous bolus infusion over 2 min (BIG group). 17 of the 58 patients in the BIG group (29.3%) and 4 of the 62 patients (6.5%) in the SIG group had akathisia (p = 0.001). There were no significant differences between the BIG and SIG groups in terms of mean headache scores (p = 0.34) and no adverse reactions in the study period. Metoclopramide successfully relieved the headache symptom(s) of patients in both the BIG and SIG groups. CONCLUSION: Slowing the infusion rate of metoclopramide is an effective strategy for the improvement of headache and reducing the incidence of akathisia in patients with vascular type headache. PMID- 17916878 TI - ABCG1 and ABCG4 are coexpressed in neurons and astrocytes of the CNS and regulate cholesterol homeostasis through SREBP-2. AB - Here, we describe the initial characterization of Abcg4(-/-) mice and identify overlapping functions of ABCG4 and ABCG1 in the brain. Histological examination of tissues from Abcg4(+/-)/nlsLacZ and Abcg1(+/-)/nlsLacZ mice demonstrates that coexpression of Abcg4 and Abcg1 is restricted to neurons and astrocytes of the central nervous system (CNS). Interestingly, Abcg4 mRNA is undetectable outside the CNS, in contrast with the broad tissue and cellular expression of Abcg1. We also used primary astrocytes, microglia, neurons, and macrophages to demonstrate that the expression of Abcg1, but not Abcg4, is induced after the activation of liver X receptor. Cellular localization studies demonstrated that both proteins reside in RhoB-positive endocytic vesicle membranes. Furthermore, overexpression of either ABCG1 or ABCG4 increased the processing of sterol-regulatory element binding protein 2 (SREBP-2) to the transcriptionally active protein, thus accounting for the observed increase in the expression of SREBP-2 target genes and cholesterol synthesis. Consistent with these latter results, we show that the expression levels of the same SREBP-2 target genes are repressed in the brains of Abcg1(-/-) and, to a lesser extent, Abcg4(-/-) mice. Based on the results of the current study, we propose that ABCG1 and ABCG4 mediate the intracellular vesicular transport of cholesterol/sterols within both neurons and astrocytes to regulate cholesterol transport in the brain. PMID- 17916880 TI - Comings and goings, hail and farewell! PMID- 17916879 TI - 20-HETE inhibits the proliferation of vascular smooth muscle cells via transforming growth factor-beta. AB - 20-Hydroxyeicosatetraenoic acid (20-HETE), a cytochrome P450 arachidonic acid metabolite, has been shown to modulate the growth of vascular smooth muscle cells (VSMCs). We asked whether 20-HETE modulates the proliferation of R22D cells, a clonal VSMC from neonatal rats, by releasing transforming growth factor-beta (TGF beta). Incubation of R22D cells with 20-HETE for 24 h attenuated [(3)H]thymidine incorporation in a concentration-dependent manner without causing the release of lactate dehydrogenase. 20-HETE also inhibited platelet-derived growth factor (PDGF)-induced [(3)H]thymidine incorporation in R22D cells and human VSMCs. At 5 muM, 20-HETE reduced [(3)H]thymidine incorporation by 34 +/- 6%; anti-TGF-beta neutralizing antibody, but not nonspecific IgG, completely reversed the attenuated [(3)H]thymidine incorporation induced by 20-HETE. In addition, 20-HETE attenuated fetal bovine serum- and PDGF-induced expression of cyclin D1, a downstream effector of TGF-beta(1), which was reversed by anti-TGF-beta antibody. Further studies demonstrated that 20-HETE may increase TGF-beta release to a level high enough to inhibit [(3)H]thymidine incorporation without altering the steady-state mRNA level of TGF-beta. Nevertheless, pretreatment of indomethacin (a cyclooxygenase inhibitor) or paxilline (a potassium channel inhibitor) did not affect the inhibitory effect on DNA synthesis induced by 20-HETE. These results demonstrate for the first time a growth-inhibitory effect induced by 20-HETE, which may be mediated by TGF-beta. PMID- 17916881 TI - Protecting children from dangerous products: fundamental change needed. PMID- 17916882 TI - Evidence-based practice in road casualty reduction. PMID- 17916883 TI - Overdoses are injuries too. PMID- 17916884 TI - Obesity and workplace traumatic injury: does the science support the link? AB - OBJECTIVE: To explore whether obesity is associated with non-fatal traumatic occupational injury. DESIGN: Systematic literature review. METHODS: The peer reviewed literature was searched from 1 January 1980 to 31 December 2005 for studies on the risk of overweight and obesity on non-fatal traumatic occupational injuries among non-office employees. The search was conducted using Medline, eLCOSH, NIOSHTIC-2, CINAHL, PsycLit, and OSH-ROM. Studies were excluded that focused on military populations, chronic/repetitive workplace injuries, back pain, only height as a risk factor, or were not written in English. RESULTS: The search identified only 12 studies. The risk of injury for obese versus non-obese employees overall was slightly increased, although many of the estimates were not statistically significant. In studies in which increased risk estimates were shown, there was limited exploration of the mechanism of obesity-related injury, but the influence of chronic disease, fatigue or sleepiness, ergonomics, and physical limitations were most often hypothesized. DISCUSSION: With the current growing prevalence of obesity worldwide, more research is needed to better establish its impact on workplace injuries and lost work time. Studies are needed that use large diverse samples, advanced statistical methods, and control for potential confounders, and explore issues related to temporality. Gaining a better understanding of how obesity influences workplace injury may foster the development of interventions that address weight, while still emphasizing the important environmental and sociocultural risk factors for injury. PMID- 17916885 TI - Smooth handling: the lack of safety-related consumer information in car advertisements. AB - OBJECTIVE: To examine the content and trends of safety-related consumer information in magazine vehicle advertisements, as a case study within the worldwide marketing of vehicles. METHODS: Content analysis of popular current affairs magazines in New Zealand for the 5-year period 2001-2005 was undertaken (n = 514 advertisements), supplemented with vehicle data from official websites. RESULTS: Safety information in advertisements for light passenger vehicles was relatively uncommon with only 27% mentioning one or more of nine key safety features examined (average: 1.7 out of nine features in this 27%). Also included were potentially hazardous features of: speed imagery (in 29% of advertisements), power references (14%), and acceleration data (4%). The speed and power aspects became relatively more common over the 5-year period (p < 0.05 for trend). CONCLUSIONS: To enhance informed consumer choice and improve injury prevention, governments should consider regulating the content of vehicle advertisements and vehicle marketing - as already occurs with many other consumer products. PMID- 17916886 TI - Prevention of motor-vehicle deaths by changing vehicle factors. AB - OBJECTIVE: To estimate the effect of changing vehicle factors to reduce mortality in a comprehensive study. Design/ METHODS: Odds of death in the United States during 2000-2005 were analyzed, involving specific makes and models of 1999-2005 model year cars, minivans, and sport utility vehicles using logistic regression after selection of factors to be included by examination of least-squares correlations of vehicle factors to maximize independence of predictors. Based on the regression coefficients, percentages of deaths preventable by changes in selected factors were calculated. Correlations of vehicle characteristics to environmental and behavioral risk factors were also examined to assess any potential confounding. RESULTS: Deaths in the studied vehicles would have been 42% lower had all had electronic stability control (ESC) systems. Improved crashworthiness as measured by offset frontal and side crash tests would have produced an additional 28% reduction, and static stability improvement would have reduced the deaths 11%. Although weight-power that reduces fuel economy is associated with lower risk to drivers, it increases risk of deaths to pedestrians and bicyclists but has an overall minor effect compared to the other factors. CONCLUSION: A large majority of motor-vehicle-related fatalities could be avoided by universal adoption of the most effective technologies. PMID- 17916887 TI - Suicide-associated comorbidity among US males and females: a multiple cause-of death analysis. AB - OBJECTIVE: To describe physical and mental comorbidity among male and female suicides in the US. This research replicates a seminal Australian study, which permits inference on comparative cause-of-death data quality. DESIGN AND SETTING: National cross-sectional study of suicides and unintentional injury decedents (comparison group) using Multiple Cause of Death (MCOD) public use files for 1999 2003. MAIN OUTCOME MEASURES: Prevalence of medical conditions; relative odds of suicide-associated comorbidity. RESULTS: For 14% of male suicides and 19% of female suicides, comorbidity was shown on their death certificates. Respective prevalences for mental and physical comorbidity were 7.2% and 7.1% for males and 10.0% and 9.3% for females. Mean numbers of psychopathologies mentioned were virtually identical for male and female suicides (1.22 and 1.21) for whom comorbidity was registered. Mean mentions of physical disease were lower for male suicides: 1.64 vs 1.77. Multiple medical conditions were registered for 4% of male suicides and 6% of female suicides. Suicides manifested excess comorbidity for depression and mood disorders (adjusted odds ratio (AOR) = 34.6, 95% CI = 32.41 to 36.92), schizophrenia (AOR = 2.5, 95% CI = 2.16 to 2.88), and cancer (AOR = 2.1, 95% CI = 1.93 to 2.19), but unexpectedly no excess comorbidity for HIV. CONCLUSIONS: The US system of death certification appears to be less fastidious than the Australian system. Although comorbidity patterns were very similar, only half as much psychopathology was reported for US suicides as for Australian suicides. A questionable deficit of comorbid physical disease was also documented for US suicides. Findings have important implications for medical training, as well as for suicide surveillance, policy, and prevention. PMID- 17916888 TI - Pediatric hospitalizations for bicycle-related injuries. AB - OBJECTIVES: To determine the incidence of bicycle-related injury hospitalizations among children and adolescents 20 years of age and younger and to examine the associated use of healthcare resources. DESIGN: Nationally representative data from the 2003 Healthcare Cost and Utilization Project's Kids' Inpatient Database (KID). OUTCOME MEASURES: National estimates of hospitalization for bicycle related injuries according to patient demographics, type of injury, total hospital charges, and length of hospital stay. RESULTS: In 2003, an estimated 10,700 children were hospitalized for a bicycle-related injury in the USA. Inpatient charges totaled nearly $200 million with a mean charge of $18,654 per hospitalization. The national rate was 12.7 hospitalizations per 100,000 children. Young adolescents aged 10-13 years accounted for the highest percentage of cases (36.6%) followed by children aged 6-9 years (25.1%). Most patients were male (76.7%) and resided in an urban area (94.4%). A head injury was diagnosed in one out of three hospitalized bicyclists; 30% were due to a motor vehicle collision. CONCLUSIONS: Pediatric bicycle-related hospitalizations are a significant public health problem. The morbidity and mortality among children and the economic costs to society are large. The patient characteristics and injury types identified by this study should be used to develop targeted prevention strategies. PMID- 17916889 TI - Area socioeconomic status and childhood injury morbidity in New South Wales, Australia. AB - OBJECTIVE: To explore the relationship between child injury morbidity and socioeconomic status. DESIGN: A cross-sectional analysis of routinely collected hospital separation data for unintentional injury for the period 1999/2000 2004/2005. SETTING: All statistical local areas of New South Wales (NSW), Australia SUBJECTS: 110 549 unintentional injury-related hospital separations for NSW children aged 0-14 years. MAIN OUTCOME MEASURE: Adjusted incidence rate ratios (IRRs) for hospital separations for unintentional injury (for all injury and by individual injury mechanisms) by quintile of socioeconomic disadvantage for children aged 0-14 years. RESULTS: There was no clear relationship between socioeconomic status and injury when all injury mechanisms were combined. However, children in the more disadvantaged quintiles were more likely to be hospitalized than children in the least disadvantaged quintile for the following injury mechanisms: motor cycle (point estimates for IRRs across the socioeconomic status quintiles ranged from 2.95 to 4.02 relative to the least disadvantaged quintile), motor-vehicle occupant (IRR range 1.33-2.27), pedestrian (IRR range 1.43-2.54 for ages 0-4 years), pedal cyclist (IRR range 1.30-1.50), fire and burns (IRR range 1.37-2.00), and poisoning (IRR range 1.32-1.91). Similarly, hospital separation rates for foreign body, other transport, and pedestrian (aged 5-9 years) injuries were also greater, but the differences were not statistically significant across all quintiles. These injury mechanisms accounted for about 25% of the hospital separations. CONCLUSIONS: The relationship between relative socioeconomic disadvantage and injury risk in NSW children is strongest for transport-related injuries, fires and burns, and poisoning. Interventions that address these specific injury mechanisms may help to reduce the disparity between high and lower socioeconomic groups. PMID- 17916890 TI - Investigating the over-representation of older persons in do-it-yourself home maintenance injury and barriers to prevention. AB - OBJECTIVE: To examine why older persons undertake high-risk do-it-yourself (DIY) home maintenance and under what circumstances, what constitutes acceptable low risk alternatives to DIY, and to assess if alternatives are feasible in the current context. DESIGN: Exploratory qualitative study using focus-group methodology. SETTING AND SUBJECTS: Fifteen focus groups were conducted, involving 118 persons aged 60 years and older, in two Melbourne communities. Participants resided locally, participated in local seniors groups, or received treatment for a DIY injury at one of two public hospitals serving these communities. RESULTS: Older persons' involvement in DIY ranged from necessity to choice. A number chose DIY for general fitness enhancement, satisfaction and pride in a job well done, and giving meaning and enjoyment to daily tasks. However, some older, frailer seniors were forced into DIY because of difficulties in choosing appropriate alternatives; lack of knowledge of some available resources and services; the challenge of accessing cost-effective and reliable private service providers; and fear of vulnerability to overcharging, overservicing or their personal security. Preferred DIY alternatives were local government providers, local paper advertised services, recommendations to private service providers and family, friends or neighbors. Lack of knowledge of other existing alternatives was an impediment to preventing DIY injury, or accessing DIY alternatives. A number of potentially feasible alternatives to DIY were identified from our review. CONCLUSIONS: This research is an important first step in understanding issues facing community-dwelling seniors remaining at home, and provides a basis on which government agencies and other providers can develop services to meet increasing needs. PMID- 17916891 TI - Effects of the Occupational Safety and Health Administration's control of hazardous energy (lockout/tagout) standard on rates of machinery-related fatal occupational injury. AB - OBJECTIVE: To evaluate the impact of the United States' federal Occupational Safety and Health Administration's control of hazardous energy (lockout/tagout) standard on rates of machinery-related fatal occupational injury. The standard, which took effect in 1990, requires employers in certain industries to establish an energy control program and sets minimum criteria for energy control procedures, training, inspections, and hardware. DESIGN: An interrupted time series design was used to determine the standard's effect on fatality rates. Machinery-related fatalities, obtained from the National Traumatic Occupational Fatalities surveillance system for 1980 through 2001, were used as a proxy for lockout/tagout-related fatalities. Linear regression was used to control for changes in demographic and economic factors. RESULTS: The average annual crude rate of machinery-related fatalities in manufacturing changed little from 1980 to 1989, but declined by 4.59% per year from 1990 to 2001. However, when controlling for demographic and economic factors, the regression model estimate of the standard's effect is a small, non-significant increase of 0.05 deaths per 100 000 production worker full-time equivalents (95% CI -0.14 to 0.25). When fatality rates in comparison groups that should not have been affected by the standard are incorporated into the analysis, there is still no significant change in the rate of machinery-related fatalities in manufacturing. CONCLUSIONS: There is no evidence that the lockout/tagout standard decreased fatality rates relative to other trends in occupational safety over the study period. A possible explanation is voluntary use of lockout/tagout by some employers before introduction of the standard and low compliance by other employers after. PMID- 17916892 TI - Child drowning deaths in Xiamen city and suburbs, People's Republic of China, 2001 5. AB - BACKGROUND: Risk-factor research and prevention programs targeting drowning deaths in children have been well developed in industrialized countries, but little research has been undertaken in developing countries where the majority of drowning deaths occur. We conducted an epidemiological study to describe the sociodemographic characteristics, drowning circumstances, and medical service in drowning deaths of children in Xiamen city and suburbs, People's Republic of China. MATERIAL AND METHODS: Drowning deaths in 1-14-year-old children between 2001 and 2005 were identified using death certificates. Parents of each case were interviewed face to face about the sociodemographics of the family and child, the drowning event, and medical care received. Mortalities were calculated using census data for urban and rural areas, and Poisson regression was used to evaluate confounding effects and interactions of several major risk factors for drowning death. RESULTS: Of 67 drowning deaths identified, 52 (77.6%) were males. A higher proportion of deaths were in children aged 5-9 years (40.3%) and 10-14 years (40.3%). The drowning mortality per 100 000 population was 5.84 in rural areas and 0.75 in urban areas. Drowning events occurred most commonly during the summer months (56.7% from June to August), during the hours of 13:00-17:59 (62.7%), and in natural or man-made bodies of water (eg, ponds, ditches, construction sites, and wells). None of the children were proficient swimmers, the majority of drowning events (88.1%) occurred in the absence of adult supervision, and 86.6% children died at the scene without any medical care. Results from muiltivariable Poisson regression analysis indicated that 10-14-year old boys were at the highest risk of drowning deaths in this area. DISCUSSION AND CONCLUSIONS: Drowning deaths in children in Xiamen city and suburbs follow trends that are markedly different from patterns observed in other countries. Different prevention strategies may be required for preventing child drowning deaths in Xiamen and other developing regions. PMID- 17916893 TI - Intracluster correlation coefficient in multicenter childhood trauma studies. AB - OBJECTIVE: To calculate the intracluster correlation coefficient (ICC) for emergency department (ED) shock rate, early trauma death (ie, death during the first 24 h after arrival at hospital), and in-hospital trauma death rate for multicenter childhood injuries. METHODS: The National Trauma Data Bank (5th revision), the largest multicenter trauma registry in the US, was used. Data from 80 trauma centers were used to calculate the ICC for in-hospital trauma death rate. Thirty three states provided data for calculation of the ICC for ED shock and early trauma death rate. RESULTS: From 2000 to 2004, 13% of the 952 242 patients in the National Trauma Data Bank were <15 years old. Approximately 17 000 of these children had injuries with an injury severity score >15, of whom 84% (14 095 subjects) were hospitalized at 80 level I or II trauma centers in 33 states. The ICCs for ED shock rate, early trauma death rate, and in-hospital death rate were 0.005 (95% CI 0.000 to 0.010), 0.014 (95% CI 0.004 to 0.024), and 0.023 (95% CI 0.013 to 0.033), respectively. These ICCs were calculated for boys and girls and also for blunt and penetrating injuries. CONCLUSION: Clustered childhood trauma studies that aim to compare different aspects of pre-hospital and hospital trauma care should incorporate these ICCs for sample calculation. When cluster randomized clinical trials are mounted, if sample sizes are calculated without adjustment for ICC, then the planned trial is likely to be seriously underpowered. PMID- 17916895 TI - Providing car seat checks with well-child visits at an urban health center: a pilot study. AB - OBJECTIVE: To evaluate a pilot program of providing child restraint system (CRS) checks by certified technicians with well-child care in an urban health center serving a low-income community. METHODS: During well-child care, nationally certified child passenger safety technicians assessed CRS use, educated care givers, corrected misuse, and provided a new CRS if necessary. The program's effect was assessed at a subsequent medical visit. RESULTS: A total of 3650 CRS checks were performed. CRS non-use was found for 307 (17%) infants, 604 (50%) toddlers, and 593 (88%) booster seat-sized children. Exposure to the program was associated with a significant positive effect on CRS use (p<0.001) and significant improvements in the major components of misuse (p<0.05) months later. CONCLUSIONS: This urban health center has high rates of CRS non-use and near universal misuse. Providing CRS checks by certified technicians during well-child care is a promising means of promoting sustained and improved CRS use. PMID- 17916894 TI - Behavioral assessment of child-directed canine aggression. AB - OBJECTIVE: To characterize behavioral circumstances of bites to children by dogs presented to a veterinary behavior clinic. METHODS: Retrospective case series examining medical records of dogs presenting by referral to a university veterinary hospital for aggression and which had bitten a child <18 years old. Behavioral data included age of victim, familiarity with dog, and circumstances of bites. RESULTS: Records of bites to 111 children were examined. Children <6 years old were most commonly bitten in association with resource guarding (44%), whereas older children were most commonly bitten in association with territory guarding (23%). Similarly, food guarding was the most common circumstance for bites to familiar children (42%) and territory guarding for bites to unfamiliar children (53%). Behavioral screening of the 103 dogs examined revealed resource guarding (61%) and discipline measures (59%) as the most common stimuli for aggression. Anxiety screens revealed abnormalities in 77% of dogs. Potential contributory medical conditions were identified/suspected in 50% of dogs. When history before presentation was known, 66% of dogs had never previously bitten a child, and 19% had never bitten any human. Most dogs (93%) were neutered, and 66% of owners had taken their dogs to obedience training classes. CONCLUSIONS: Most children were bitten by dogs with no history of biting children. There is a high rate of behavioral abnormalities (aggression and anxiety) in this canine population. Common calming measures (neutering, training) were not routinely effective deterrents. PMID- 17916896 TI - Demographic, socioeconomic, and attitudinal associations with children's cycle helmet use in the absence of legislation. AB - The association of demographic, socioeconomic, and attitudinal factors with children's cycle-helmet use was investigated using self-reported data from 9775 children aged 8-12 in England, where there are currently no legal requirements that cyclists (of any age) wear helmets. The results suggest that demographic factors, socioeconomic status, and attitudes influence whether children wear bicycle helmets but that attitudinal factors are most important. PMID- 17916897 TI - Finding studies for inclusion in systematic reviews of interventions for injury prevention the importance of grey and unpublished literature. PMID- 17916899 TI - The alarm anti-protease, secretory leukocyte protease inhibitor, is a proliferation and survival factor for ovarian cancer cells. AB - Alarm anti-proteases are secreted locally in response to inflammation and have been shown to be elevated in cancers. Secretory leukocyte protease inhibitor (SLPI), an alarm anti-protease, is amplified in ovarian carcinoma and is induced and binds to and protects progranulin (prgn) in inflammation. We reported prgn is a survival protein in ovarian cancer and now hypothesize that SLPI/prgn would promote proliferation and survival. Neutralizing anti-SLPI antibody treatment of HEY-A8 and OVCAR3 ovarian cancer cells decreased cell number (P < 0.001), induced apoptosis and reduced prgn quantity. This was confirmed using SLPI small interfering RNA. Prgn and SLPI were co-immunoprecipitated and co-localized by confocal microscopy. Prgn is a substrate of the serine protease elastase and SLPI is an inhibitor of elastase. Elastase reduced prgn expression, inhibited proliferation in a dose-dependent manner (P G) and IL6 (rs1800795, -174G>C) polymorphisms were significantly associated with risk of glioma in the pooled analysis (P trend = 0.006 and 0.04, respectively), although these became attenuated after controlling for the false discovery rate (P trend = 0.07 and 0.22, respectively). Our results underscore the importance of pooled analyses in genetic association studies and suggest that SNPs in cytokine genes may influence susceptibility to glioma. PMID- 17916901 TI - 3-Morpholinopropyl isothiocyanate is a novel synthetic isothiocyanate that strongly induces the antioxidant response element-dependent Nrf2-mediated detoxifying/antioxidant enzymes in vitro and in vivo. AB - The induction of NF-E2-related factor-2 (Nrf2)-mediated detoxifying/antioxidant enzymes is recognized as an effective strategy for cancer chemoprevention. Here, we report that 3-morpholinopropyl isothiocyanate (3MP-ITC) is an exceptionally strong chemical inducer of these enzymes. Exposure of 3MP-ITC in HepG2C8 cells not only induced endogenous Nrf2 protein but also suppressed endogenous Kelch like ECH-associated protein 1, resulting in an increased nuclear accumulation of Nrf2. Using chemical inhibitors of protein synthesis (cycloheximide) and 26S proteosomal degradation (MG-132), we observed that the induction of Nrf2 protein by 3MP-ITC appeared to be post-translationally regulated. 3MP-ITC activated ERK1/2 and JNK1/2 and the activation of antioxidant response element (ARE) by 3MP ITC was significantly attenuated by chemical inhibition of PKC and PI3K signaling pathways in HepG2C8 cells. Treatment with 3MP-ITC significantly depleted the intracellular level of glutathione (GSH) in HepG2C8 cells and oral administration of 3MP-ITC increased the protein expression of hepatic NAD[P]H:quinone oxidoreductase-1 and Nrf2 in Nrf2 (+/+) but not in Nrf2 (-/-) mice, whereas UDP glucuronosyl transferase 1A1 was induced in both genotypes. Our results indicate that 3MP-ITC is a novel ITC that strongly induces Nrf2-dependent ARE-mediated detoxifying/antioxidant enzymes in vitro and in vivo via the Nrf2 signaling pathway coupled with GSH depletion and activation of multiple signaling kinase pathways, which could be potentially useful agent for cancer chemoprevention. PMID- 17916902 TI - NDRG1, a growth and cancer related gene: regulation of gene expression and function in normal and disease states. AB - N-myc downstream-regulated gene 1 (NDRG1) is an intracellular protein that is induced under a wide variety of stress and cell growth-regulatory conditions. NDRG1 is up-regulated by cell differentiation signals in various cancer cell lines and suppresses tumor metastasis. Despite its specific role in the molecular cause of Charcot-Marie-Tooth type 4D disease, there has been more interest in the gene as a marker of tumor progression and enhancer of cellular differentiation. Because it is strongly up-regulated under hypoxic conditions, and this condition is prevalent in solid tumors, its regulation is somewhat complex, governed by hypoxia-inducible factor 1 alpha (HIF-1alpha)- and p53-dependent pathways, as well as its namesake, neuroblastoma-derived myelocytomatosis, and probably many other factors, at the transcriptional and translational levels, and through mRNA stability. We survey the data for clues to the NDRG1 gene's mechanism and for indications that the NDRG1 gene may be an efficient diagnostic tool and therapy in many types of cancers. PMID- 17916903 TI - Comparative genomics of susceptibility to mammary carcinogenesis among inbred rat strains: role of reduced prolactin signaling in resistance of the Copenhagen strain. AB - To elucidate the molecular basis for differential susceptibilities to mammary carcinogenesis, we compared the transcriptomes of normal mammary glands from pubescent female rats of the resistant Copenhagen (Cop) strain with those of the susceptible Fischer 344 (F344), August x Copenhagen Irish (ACI), Buffalo/N (Buf/N), Wistar-Furth (WF) strains and F1 (Cop x F344) progeny (F1). Gene expression profiles in mammary tissue within each rat strain were remarkably similar, indicating that gene expression was determined by genetic background. We next identified the subset of genes that were differentially expressed in all susceptible strains relative to the resistant Cop strain. Among these, the messenger RNAs encoding prolactin (Prl) and its cell surface receptor were significantly elevated in all susceptible strains. The expression levels of several Prl-regulated genes were also significantly elevated, indicating the presence of increased Prl signaling in mammary glands of all susceptible strains. Pathway analysis of gene expression profiles further identified the Prl-activated Jak/STAT-signaling pathway among the pathways that most distinguished sensitive rat strains from the resistant Cop rat. To test the hypothesis that reduced levels of the Prl signaling in mammary tissue partially contributed to the genetic resistance to mammary carcinogenesis, we used the neuroleptic drug, perphenazine, to transiently elevate serum Prl levels in the Cop strain. Whereas Cop rats are resistant to N-nitroso-N-methylurea (NMU)-induced mammary carcinogenesis, approximately 5% of pubescent Cop females treated with perphenazine and NMU exposure developed mammary adenocarcinomas with latencies comparable with those of sensitive strains. Together, these finding indicated that in the rat, the molecular mechanisms underlying genetic susceptibility to mammary carcinogenesis include de-regulation of Prl signaling. PMID- 17916904 TI - Decreased 7,12-dimethylbenz[a]anthracene-induced carcinogenesis coincides with the induction of antitumor immunities in adult female B6C3F1 mice pretreated with genistein. AB - The objective of this study was to determine if genistein (GEN) modulation of the immune responses might contribute to the increased host resistances to tumors. A time-course study was performed in adult female B6C3F1 mice that had been exposed to GEN for 1-4 weeks at the dose level of 20 mg/kg by gavage. A significant increase in ex vivo cytotoxic T lymphocyte (CTL) activity was observed in the periods of 2 weeks and 4 weeks. Moreover, increased activities of CTLs were associated with a decrease in the percentage of CD4(+)CD25(+) T cells and an increase in the production of interferon-gamma and activation of STAT1 (signal transducer and activator of transcription 1) and STAT4. Additionally, exposure of mice to GEN increased the activities of in vivo CTLs. An increased activity of natural killer (NK) cells was also observed. Further study in the B16F10 tumor model suggested that GEN-mediated enhancement in host resistance to B16F10 tumor was partially related to its potentiating effect on NK cells. Finally, 7,12 dimethylbenz[a]anthracene (DMBA)-induced tumor model was employed to determine the chemopreventive effect of oral GEN treatment. Mice pretreated with GEN for 2 weeks by gavage, the time when an enhanced CTL activity had been produced, had a decreased susceptibility toward DMBA-mediated carcinogenesis, while treatment with GEN after tumor induction conferred no protection. In conclusion, pretreatment with GEN by gavage could enhance host resistances to the B16F10 tumor and DMBA-induced carcinogenesis, suggesting that GEN modulation of immune response was, at least partially, responsible for the antitumor effect of this compound. PMID- 17916905 TI - Polymorphisms of GSTP1 and GSTT1, but not of CYP2A6, CYP2E1 or GSTM1, modify the risk for esophageal cancer in a western population. AB - Esophageal cancer is among the most common and fatal tumors in the world. Eighty percent of esophageal tumors are esophageal squamous cell carcinoma (ESCC). Brazil is one of the high incidence areas in the West, where tobacco and alcohol consumption have been associated with ESCC. However, polymorphisms in xenobiotic metabolizing genes may also contribute to the risk. Therefore, in this study, we analyzed the risk of ESCC associated with tobacco and alcohol consumption and with polymorphisms of CYP2A6 (CYP2A6*2), CYP2E1 (CYP2E1*5B, CYP2E1*6), GSTP1 (Ile105Val), GSTM1 and GSTT1 null genotypes in 126 cases and 252 age- and gender matched controls. Data on the amount, length and type of tobacco and alcohol consumed were collected, and DNA was extracted from blood lymphocytes from all individuals. Polymorphisms were analyzed by polymerase chain reaction (PCR) multiplex (GSTM1 and T1), PCR-Restriction Fragment Length Polymorphism (CYP2E1*5B and *6 and GSTP1 Ile105Val) or allele-specific PCR amplification (CYP2A6*2). Risks were evaluated by multivariate conditional regression analysis. As expected, tobacco [odds ratio (OR) = 6.71, 95% confidence interval (95% CI) 3.08 14.63] and alcohol (OR = 16.98, CI 7.8-36.98) consumption, independently or together (OR = 26.91, CI 13.39-54.05) were risk factors. GSTP1 Ile105Val polymorphism was an independent risk factor (OR = 2.12, CI 1.37-3.29), whereas GSTT1 wild-type was an independent protective factor for ESCC (OR = 0.37, CI 0.16 0.79). There was approximately 80% statistical power to detect both results. There was no risk associated with CYP2A6, CYP2E1 and GSTM1 polymorphisms. In conclusion, this study suggests an opposite role of GSTP1 and GSTT1 polymorphisms for the risk for ESCC. PMID- 17916906 TI - GRB-7 facilitates HER-2/Neu-mediated signal transduction and tumor formation. AB - Growth factor receptor-bound protein-7 (GRB-7), an adaptor molecule, can interact with multiple signal transduction molecules. GRB-7 is amplified concurrently with HER-2/Neu in most, if not all, of breast cancer with chromosome 17q11-21 amplification. GRB-7 gene amplification is associated with RNA over-expression. We show GRB-7 protein is over-expressed by immunoblotting in breast cancer cell lines and primary breast tumors with HER-2/Neu protein over-expression. Over expression of GRB-7 in MCF-7 breast cancer cells that over-express HER-2/Neu leads to activation of tyrosine phosphorylation of HER-2/Neu. Knockdown of GRB-7 expression in SKBR-3 breast cancer cells with naturally occurring HER-2/Neu gene amplification decreases tyrosine phosphorylation of HER-2/Neu. Activation of HER 2/Neu phosphorylation is associated with increase in tyrosine phosphorylation of phosphoinositide-specific lipase C-gamma-1 (PLC-gamma-1) and recruitment of PLC gamma-1 to HER-2/Neu protein molecule. Activation of downstream protein kinase C (PKC) pathway is evidenced by increase in the phosphorylation of a common PKC substrate-myristoylated alanine-rich protein kinase C substrate (MARCKS). In addition, over-expression of GRB-7 in MCF-7 breast cancer cells that over-express HER-2/Neu leads to activation of AKT phosphorylation. Knockdown of GRB-7 expression in MB-453 and SKBR-3 breast cancer cells results in decrease in AKT phosphorylation. GRB-7 over-expression therefore facilitates activation of phosphorylation of HER-2/Neu and AKT in breast cancer cells with HER-2/Neu over expression. GRB-7 over-expression in MCF-7 cells over-expressing HER-2/Neu leads to morphologic change of cells and promotes tumor xenograft growth in nude mice. GRB-7 over-expression therefore plays pivotal roles in activating signal transduction and promoting tumor growth in breast cancer cells with chromosome 17q11-21 amplification. PMID- 17916907 TI - Involvement of matrix metalloproteinase-9 in stromal cell-derived factor-1/CXCR4 pathway of lung cancer metastasis. AB - Lung caner cells have a striking tendency to metastasize to bone. The chemokine stromal cell-derived factor-1 (SDF-1) is constitutively secreted by osteoblasts and bone marrow stromal cells and plays a key role for homing of hematopoietic cells to the bone marrow. Reverse transcriptase-polymerase chain reaction and flow cytometry studies demonstrated SDF-1 receptor (CXCR4) messenger RNA (mRNA) and surface expression of CXCR4 in lung cancer cell lines, especially higher in those with high invasiveness (A549) as compared with lower level in H928 cells and H1299 cells. SDF-1, osteoblast-conditioned medium (OBCM) and stromal cell conditioned medium all induced the invasiveness of lung cancer cells. Matrix metalloproteinase (MMP)-9 small interfering RNA inhibited the SDF-1alpha- or OBCM induced MMP-9 expression and thereby significantly inhibited the SDF-1alpha- or OBCM-induced cell invasion. Furthermore, mitogen-activated protein kinase kinase inhibitor PD98059 suppressed SDF-1alpha-induced MMP-9 mRNA expression. Transient transfection with dominant-negative extracellular signal-regulated kinase (ERK) mutant also showed that the ERK-signaling pathway was involved in SDF-1alpha induced MMP-9 expression. Moreover, nuclear factor-kappaB (NF-kappaB) decoy oligodeoxynucleotide suppressed the MMP-9 promoter activity enhanced by SDF 1alpha. Both mitogen-activated protein kinase kinase inhibitor and ERK mutant also antagonized SDF-1alpha-induced NF-kappaB-driven luciferase promoter activity. Taken together, our results indicated that bone marrow-derived-SDF 1alpha enhances the invasiveness of lung cancer cells by increasing MMP-9 expression through the CXCR4/ERK/NF-kappaB signal transduction pathway. PMID- 17916909 TI - Silibinin suppresses in vivo growth of human prostate carcinoma PC-3 tumor xenograft. AB - Chemoprevention is an upcoming approach to control cancer including prostate cancer (PCa). Here, we studied the efficacy and associated mechanisms of a chemopreventive agent silibinin against ectopically growing and established advanced human prostate carcinoma PC-3 tumor xenografts in athymic nude mice. Dietary silibinin (0.5%, w/w) did not show any adverse health effect in mice. In first protocol, silibinin started 1 week prior to xenograft implantation and continued for 60 additional days, whereas in the second protocol, silibinin treatment was started after 25 days of established tumors for 4, 8 and 16 days. Silibinin inhibited tumor growth rate in both protocols showing up to 35% (P = 0.010) and 18-56% (P = 0.002 to <0.001) decrease in tumor volume per mouse and 27% (P < 0.01) and 44% (P = 0.014) decrease in tumor weight per mouse, respectively. In first protocol, silibinin decreased (P < 0.001) tumor cell proliferation and microvessel density but increased (P < 0.001) apoptosis. An increase in insulin-like growth factor-binding protein-3 (IGFBP-3) expression with a concomitant decrease in vascular endothelial growth factor (VEGF) expression was noted. Silibinin strongly increased phospho-extracellular signal regulated kinase 1/2 (ERK1/2), Cip1/p21 and Kip1/p27 (cyclin-dependent kinase inhibitors) levels but moderately decreased Bcl-2 and survivin levels. In established tumors, similar biomarkers and molecular changes were observed due to silibinin corresponding to its antitumor efficacy. These findings identified in vivo antitumor efficacy of silibinin against PC-3 human PCa in both intervention protocols accompanied with its anti-proliferative, pro-apoptotic and anti angiogenic activities. At molecular level, silibinin increased IGFBP-3, Cip1/p21, Kip1/p27 levels and ERK1/2 activation and decreased Bcl-2, survivin and VEGF levels in tumors. PMID- 17916910 TI - Exposure of Fischer 344 rats to a weak power frequency magnetic field facilitates mammary tumorigenesis in the DMBA model of breast cancer. AB - The possibility that long-term exposure to relatively weak power frequency magnetic fields (MFs) emanating from the generation, transmission and use of electricity could increase the risk of breast cancer is a matter of ongoing debate. Laboratory studies using well-defined exposure conditions are useful to examine whether exposure to MF affects mammary tumorigenesis. Previous studies from different laboratories using the 7,12-dimethylbenz[a]anthracene (DMBA) model of breast cancer in female Sprague-Dawley (SD) rats have been inconclusive, which has been related to differences in MF sensitivity between SD substrains used in these studies. When we compared the effects of MF exposure on cell proliferation in the mammary gland of various outbred and inbred rat strains, Fischer 344 was the only inbred strain that exhibited a marked increase in cell proliferation. Based on these data, we suggested that MF exposure should significantly facilitate development and growth of mammary tumors in Fischer 344 rats, which was tested in the present study. Groups of 108 DMBA-treated rats were either MF exposed (100 muT, 50 Hz) or sham exposed for 26 weeks. MF exposure significantly facilitated mammary tumorigenesis. The incidence of rats with grossly recorded, histologically verified adenocarcinomas was increased by 45% (P = 0.0095). The most pronounced MF effect on tumor incidence was seen in the cranial inguinal complexes (L/R5). These data indicate that Fischer 344 rats are a suitable inbred strain to study the mechanisms underlying the effects of MF exposure on mammary tumorigenesis. PMID- 17916908 TI - Survivin repression by p53, Rb and E2F2 in normal human melanocytes. AB - The inhibitor of apoptosis protein survivin is a dual mediator of apoptosis resistance and cell cycle progression and is highly expressed in cancer. We have shown previously that survivin is up-regulated in melanoma compared with normal melanocytes, is required for melanoma cell viability, and that melanocyte expression of survivin predisposes mice to ultraviolet-induced melanoma and metastasis. The mechanism of survivin up-regulation in the course of melanocyte transformation and its repression in normal melanocytes, however, has not been clearly defined. We show here that p53 and retinoblastoma (Rb), at basal levels and in the absence of any activating stimuli, are both required to repress survivin transcription in normal human melanocytes. Survivin repression in melanocytes does not involve alterations in protein stability or promoter methylation. p53 and Rb (via E2Fs) regulate survivin expression by direct binding to the survivin promoter; p53 also affects survivin expression by activating p21. We demonstrate a novel role for E2F2 in the negative regulation of survivin expression. In addition, we identify a novel E2F-binding site in the survivin promoter and show that mutation of either the p53- or E2F-binding sites is sufficient to increase promoter activity. These studies suggest that compromise of either p53 or Rb pathways during melanocyte transformation leads to up regulation of survivin expression in melanoma. PMID- 17916911 TI - High red meat diets induce greater numbers of colonic DNA double-strand breaks than white meat in rats: attenuation by high-amylose maize starch. AB - Human population studies show that dietary red and processed, but not white, meats are associated with increased risk of colorectal cancer but dietary fibre appears to be protective. We examined whether dietary cooked red or white meat had differential effects on colonic DNA damage in rats and if resistant starch (RS), a dietary fibre component, provided protection. Rats were fed diets containing approximately 15, 25 or 35% of cooked beef or chicken, both with or without 20% high-amylose maize starch (HAMS) as a source of RS, for 4 weeks. DNA single-strand breaks (SSB) and double-strand breaks (DSB) were measured in isolated colonocytes (by comet assay) along with apoptosis levels, colonic mucus thickness and large bowel short-chain fatty acids (SCFA). Both red and white meat increased colonocyte SSB and DSB dose dependently but damage was substantially greater with red meat. Dietary HAMS prevented these increases. Apoptotic cell numbers were increased dose dependently by red meat irrespective of HAMS feeding, whereas white meat only increased apoptotic cell numbers in the presence of HAMS. Red meat induced greater colonic mucus layer thinning than white meat but HAMS was protective in both cases. HAMS induced increases in large bowel SCFA, including butyrate, and significantly lowered concentrations of phenols and cresols. We have demonstrated that dietary red meat causes greater levels of colonic DNA SSB and DSB than white meat, consistent with the epidemiological data. Dietary RS protects against this damage and also against loss of the mucus barrier, probably through increased butyrate production. PMID- 17916912 TI - Inclusion in physical education: a review of the literature from 1995-2005. AB - The purpose of the review is to critically analyze English-written research articles pertaining to inclusion of students with disabilities in physical education published in professional journals both within and outside of the United States from 1995-2005. Each study included in this review had to meet seven a priori criteria. Findings of the 38 selected studies were divided into six focus areas: (a) support, (b) affects on peers without disabilities, (c) attitudes and intentions of children without disabilities, (d) social interactions, (e) ALT-PE of students with disabilities, and (f) training and attitudes of GPE teachers. Recommendations for future practice and research are embedded throughout the article. PMID- 17916914 TI - Moving beyond the stigma: the impression formation benefits of exercise for individuals with a physical disability. AB - This study examined whether the positive impressions formed by others of able bodied exercisers extend to people with a physical disability. Participants (226 women and 220 men) read a description of a man or woman with a spinal cord injury who was described as an exerciser, nonexerciser, or control, and then rated the target (i.e., the person being described in the vignette) on 17 personality and 9 physical dimensions. Results revealed significantly more favorable ratings for the exerciser than both the nonexerciser and control on almost all dimensions. Additionally, the male control target was rated more favorably than the female counterpart on three personality and two physical attributes. Evidently, the exerciser stereotype may undermine negative impressions of people with physical disabilities. PMID- 17916913 TI - Physical performance of individuals with intellectual disability: a 30 year follow up. AB - Physical performance of Finnish adolescents (33 females, 44 males) with moderate intellectual disability (ID) was studied over a 30 year period. This study is an extension of Lahtinen's previous work on documenting the performance of individuals with intellectual disabilities over time. This study consisted of analyzing data from a total of four data collection periods (1973, 1979, 1996 and 2003 in which participants ranged in age from 11-16, 17-22, 34-39 and 41-46 years old, respectively. Improvement from early to late adolescence, and decline during adulthood in abdominal strength/endurance, static balance, and manual dexterity were identified. The male adults with ID were moderately overweight (BMI), but the females with ID were obese. The IQ effect was significant on balance and manual dexterity. The gender differences in adulthood were significant, but differences were not noted for DS when controlling for IQ. PMID- 17916915 TI - Psychometric properties and analysis of the physical activity Social Influence Scale for adults with traumatic brain injuries. AB - The study consisted of (a) assessing the validity and reliability of the Social Influence Scale (SIS) for adults with brain injuries and (b) examining the kinds and sources of social influence and demographic variables that affect participants' physical activity behaviors. Three confirmatory factor analyses, intraclass correlations, and a repeated measures MANOVA were completed on responses from 402 adults with brain injuries. Results provide evidence of the validity and reliability of the SIS, and post hoc analyses revealed that participants reported receiving different types of social influence from family, friends, and caregivers. Findings have implications for specialists and suggestions are provided on ways to enhance the physical activity participation of adults with brain injuries. PMID- 17916916 TI - Effects of sport participation on the basketball skills and physical self of adolescents with conduct disorders. AB - The purpose of this study was to examine the long-term effects of sport participation on the basketball skills and physical self-concept of adolescents with conduct disorders (CD). Participants were 24 adolescent males with CD, divided equally into three groups: (a) inter-establishment basketball (IEBB), (b) integrated scholastic basketball (ISBB), and (c) control-adapted physical activity (APA). The basketball skills tests and physical self-concept were both administrated 4 times over an 18-month period. Results indicated (a) an improvement in basketball skills in both competitive groups (i.e., ISBB, IEBB), (b) a significant curvilinear trend of physical self-worth scale in the three groups, and (c) no significant changes in physical self-concept in the three groups (ISBB, IEBB, and APA). In conclusion, the integrated and segregated competitive programs did not represent an effective means for improving the physical self-concept of adolescents with CD. PMID- 17916917 TI - Kinetic movement analysis in adults with vision loss. AB - The purpose of this investigation was to assess movement capabilities of adults with visual impairments in comparison to sighted peers. Thirty participants (n = 15 visually impaired; n = 15 without vision loss) were age and gender matched and assessed on three functional movement measures. A Walk Across, Forward Lunge, and Sit to Stand were completed on a long force plate and analyzed using Group MANOVAs. Individuals with visual impairment were more cautious and conservative in their movement as evident by significantly reduced performance on both the Walk Across assessment and the Forward Lunge task. Performance between groups was similar on the Sit to Stand maneuver. It was concluded that individuals with visual impairments are more cautious and have more difficulty performing tasks when their center of gravity is outside of their base of support. PMID- 17916918 TI - Accuracy of voice-announcement pedometers for youth with visual impairment. AB - Thirty-five youth with visual impairments (13.5 +/- 2.1 yrs, 13 girls and 22 boys) walked four 100-meter distances while wearing two units (right and left placement) of three brands of voice-announcement (VA) pedometers (Centrios Talking Pedometer, TALKiNG Pedometer, and Sportline Talking Calorie Pedometer 343) and a reference pedometer (NL2000). Registered pedometer steps for each trial were recorded, compared to actual steps assessed via digital video. Inter unit agreement between right and left VA pedometer placement was low (ICC range .37 to .76). A systematic error was observed for the VA pedometers on the left placement (error range 5.6% to 12.2%), while right placement VA pedometers were at or below +/- 3% from actual steps (range 2.1% to 3.3%). The reference pedometer was unaffected by placement (ICC .98, error approximately 1.4%). Overall, VA pedometers demonstrated acceptable accuracy for the right placement, suggesting this position is necessary for youth with visual impairments. PMID- 17916919 TI - Accuracy of pedometer steps and time for youth with disabilities. AB - The purpose of the study was to examine the accuracy of pedometer steps and activity time (Walk4Life, WL) for youth with developmental disabilities. Eighteen youth (11 girls, 7 boys) 4-14 years completed six 80-meter self-paced walking trials while wearing a pedometer at five waist locations (front right, front left, back right, back left, middle back). Trials were video taped to determine actual steps and activity time. Time exhibited a smaller percent error in comparison to steps across locations. Apart from the front left, location had minimal influence on accuracy. The WL demonstrates acceptable accuracy for steps and activity time. PMID- 17916920 TI - Reliability of scores from physical activity monitors in adults with multiple sclerosis. AB - We examined the reliability of scores from physical activity monitors in a sample of 193 individuals with multiple sclerosis (MS) who wore a pedometer and an accelerometer for a 7-day period. There were no significant differences among days for the pedometer (p = .12) or the accelerometer (p = .15) indicating that week and weekend days can be analyzed in a single intra-class correlation (ICC) analytic model. The 7 days of monitoring yielded ICC estimates of .93 for both the pedometer and accelerometer, and a minimum of 3 days yielded a reliability of .80 for both the pedometer and accelerometer. Results indicated that physical activity monitor scores are reliable measures of physical activity for individuals with MS. PMID- 17916921 TI - Visual behavior and motor responses of novice and experienced wheelchair tennis players relative to the service return. AB - The purpose of this study was to determine visual behavior and motor responses between experienced and novice wheelchair tennis players relative to the return in tennis. Novice (n = 7) and Experienced (n = 5) wheelchair tennis players took part in the study. Two series of serves performed to the forehand and the backhand sides were examined in both groups. One series was performed in a video based setting (two dimensional) and the other one on court (three dimensional). Experienced participants focused initially on the head/shoulders and the free arm, while novice players focused on the expected ball toss area or followed the ball from the toss to the apex. Results suggest that the experienced players obtain useful information from racket-arm cues during the stroke phase. They also performed faster motor responses as well. PMID- 17916922 TI - Trace element concentration in primary liver cancers--a systematic review. AB - BACKGROUND: The incidence of primary liver cancer varies between countries. Many of the etiological factors contributing to the geographical variations in incidence are unknown. Development of hepatocellular carcinoma has been linked to levels of trace elements. This review summarizes the evidence associating HCC with trace elements. METHODS: MEDLINE, EMBASE, and CENTRAL databases were searched. Various inclusion and exclusion criteria were applied to select the articles for inclusion. Data extraction was performed using a custom designed data extraction form. RESULTS: A total of 12,344 references were identified. Duplicates, 1,597, were excluded. Clearly irrelevant references, 10,676, were excluded through reading titles and abstracts. Some references (59) were excluded by applying the exclusion criteria. Twelve studies including 646 patients and measuring iron content (8), copper content (11), zinc (9), and selenium (2) qualified for the review. Although a meta-analysis was not possible due to heterogeneity between the studies, a clear pattern of distribution of the trace elements was discernible. CONCLUSION: Iron and zinc content are lower in HCC than in surrounding tissues or normal controls. Copper content is lower in HCC than in surrounding tissues and cirrhotic controls. Epidemiological and physiological reasons for the trace element alterations should be further investigated. PMID- 17916924 TI - Effects of selenium supplementation on antioxidant defense and glucose homeostasis in experimental diabetes mellitus. AB - The objective of this study was to investigate the effects of different forms of Se supplementation on the antioxidant defense and glucose homeostasis in experimental diabetes. Sodium selenate (SS) or selenomethionine (SM) were administered (2 micromol Se kg(-1) day(-1)) via orogastric route to streptozotocine (STZ)-induced diabetic rats in addition to basal diet for 12 weeks. Glucose levels in whole blood, glutathione peroxidase (GSH-Px) activity in erythrocytes, Se and fructosamine levels in plasma were evaluated monthly. Plasma Se levels increased significantly in all diabetic groups compared to basal measurements, being more prominent in SM group [p(SM(3)/SM(0)) = 0.018]. The increase in GSH-Px activities was significant at the end of the second month in SS [p(SS(2)/SS(0)) = 0.028], whereas at the end of the third month in SM the value was lower [p(SM3/SM0) = 0.018] and the unsupplemented diabetic control (DC) groups, p(DC(3)/DC(0)) = 0.012. Glucose increased significantly only in DC group. Fructosamine increased gradually in all diabetic groups, being significant in DC and SS groups. At the end of the third month, highest fructosamine levels were observed in SS group, which were significantly higher than the SM group [p(SM/SS) = 0.010]. In conclusion, Se augmented the antioxidant defense by increasing GSH Px activity and this effect was more prominent when Se was supplemented as SM, which exerted positive effects also on glucose homeostasis. PMID- 17916923 TI - Characterization and distribution of the selected metals in the scalp hair of cancer patients in comparison with normal donors. AB - Eighteen metals were estimated in the scalp hair samples from cancer patients (n = 111) and normal donors (n = 113). Nitric acid-perchloric acid wet digestion procedure was used for the quantification of the selected metals by flame atomic absorption spectrophotometry. In the scalp hair of cancer patients, highest average levels were found for Ca (861 microg/g), followed by Na (672 microg/g), Zn (411 microg/g), Mg (348 microg/g), Fe (154 microg/g), Sr (129 microg/g), and K (116 microg/g), whereas in comparison, the dominant metals in the scalp hair of normal donors were Ca (568 microg/g), Zn (177 microg/g), Mg (154 micraog/g), Fe (110 microg/g), and Na (103 microg/g). The concentrations of Ca, Cd, Co, Cr, Fe, K, Mg, Mn, Na, Ni, Pb, Sb, Sr, and Zn were notably higher in the hair of cancer patients as compared with normal donors, which may lead to a number of physiological disorders. Strong positive correlations were found in Mn-Pb (0.83), Cd-Cr (0.82), Cd-Li (0.57), Fe-Pb (0.56), and Fe-Mn (0.55) in the hair of cancer patients whereas Na-Cd, Li-Cr, Li-Co, Co-Cd, Li-Cd, Na-Co, Na-Li, Ca-Mg and Na-Cr exhibited strong relationships (r > 0.50) in the hair of normal donors. Principal Component Analysis (PCA) of the data revealed seven PCs, both for cancer patients and normal donors, but with significantly different loadings. Cluster Analysis (CA) was also used to support the PCA results. The study evidenced significantly different pattern of metal distribution in the hair of cancer patients in comparison with normal donors. The role of trace metals in carcinogenesis was also discussed. PMID- 17916925 TI - Impact of Zn, Cu, and Fe on the activity of carbonic anhydrase of erythrocytes in ducks. AB - The impact of zinc, copper, and iron on the duck erythrocyte carbonic anhydrase (CA) activity and the hemoglobin content in vitro culture were studied. The increase of zinc or iron addition at a low level induced the rise of CA activity, and the CA activity was inhibited by zinc or iron at a high addition level. The duck erythrocyte CA was strongly inhibited by cupric ion. The inhibition constant of duck erythrocyte CA to cupric ion is about 3.5 microM. Carbonic anhydrase compared to hemoglobin is more sensitive to zinc and copper in the environment. These findings suggest that some characteristics of duck erythrocyte CA are different from both CAI and CAII of mammals. The increase of Fe addition below 8 microM in the minimal essential medium brought about the rise of CA activity and resulted in the maximum of CA activity exceeding that induced by Zn. It provided a new evidence for the role of ferrous ion in CA. PMID- 17916926 TI - Transport of intranasally instilled fine Fe2O3 particles into the brain: micro distribution, chemical states, and histopathological observation. AB - It has been demonstrated that inhaled fine (d < 2.5 microm) and ultrafine (d < 100 nm) particles produce more severe toxicity than coarse particles. Some recent data support the concept that the central nervous system (CNS) may be a target for the inhaled fine particulates. This work describes initial observation of the transport of intranasally instilled fine ferric oxide (Fe2O3) particles in animal brain. The iron micro-distribution and chemical state in the mice olfactory bulb and brain stem on day 14 after intranasal instillation of fine Fe2O3 particle (280 +/- 80 nm) suspension at a single dose of 40 mg/kg body weight were analyzed by synchrotron radiation x-ray fluorescence and x-ray absorption near-edge structure (XANES). The micro-distribution map of iron in the olfactory bulb and brain stem shows an obvious increase of Fe contents in the olfactory nerve and the trigeminus of brain stem, suggesting that Fe2O3 particles were possibly transported via uptake by sensory nerve endings of the olfactory nerve and trigeminus. The XANES results indicate that the ratios of Fe (III)/Fe (II) were increased in the olfactory bulb and brain stem. The further histopathological observation showed that the neuron fatty degeneration occurred in the CA3 area of hippocampus. Such results imply an adverse impact of inhalation of fine Fe2O3 particles on CNS. PMID- 17916927 TI - Pretreatment effect of resveratrol on streptozotocin-induced diabetes in rats. AB - This study was planned to investigate the pretreatment effect of resveratrol on streptozotocin-induced diabetic rats. The control group consisted of 10 male albino Sprague-Dawley rats, 10-12 weeks of age, weighing approximately 295 g. The first experimental group consisted of 15 albino Sprague-Dawley rats, 10-12 weeks of age, weighing approximately 305 g. This group was administered streptozotocin (55 mg/kg, intraperitoneally). The second experimental group (n = 15) was administered resveratrol (0.5 ml/day) 10 days before streptozotocin induction. A training period was performed for all groups before the experimental procedure, and systolic arterial blood pressures and heart rates were recorded daily. At the end of the 10th day, blood samples of control and experimental groups were drawn. Total nitrite, nitrite, nitrate, malondialdehyde (MDA), copper, and zinc concentrations in plasma were measured both in control and experimental groups. Additionally, superoxide dismutase, catalase activities, and copper and zinc concentrations in red cell were determined in each group. At the end of the study, increases in catalase activity, nitric oxide level, and zinc concentrations and decreases in lipid peroxidation product MDA and copper concentrations were found in the resveratrol-pretreated diabetic group when compared to the diabetic group. PMID- 17916928 TI - Resveratrol: is there any effect on healthy subject? AB - This preliminary study was planned to investigate the effects of resveratrol on oxidative-nitrosative stress markers and on trace element concentrations in blood and on circulatory system parameters in rats. Twenty-five Sprague-Dawley male rats, 10-12 weeks old, with mean body weight of 295 g were used in the study. Administration of resveratrol (0.5 ml/day) was performed in experimental group in 10 days. In control (n = 10) and in experimental groups (n = 15), after 1 week training period, systolic arterial blood pressures and heart rates were recorded daily. At the end of the tenth day, blood samples of control and experimental groups were drawn. Total nitrite, nitrite, nitrate, malondialdehyde, copper, zinc concentrations in plasma, superoxide dismutase, and catalase activities and copper, zinc concentrations in red cell were determined both in control and experimental groups. Alterations in oxidative and nitrosative stress markers, trace element concentrations, and circulatory system parameters in experimental group compared to controls were observed. The results of this study were discussed according to the effect of resveratrol. PMID- 17916929 TI - Effect of homocysteine on bone mineral density of rats. AB - The relationship between plasma levels of homocysteine (Hcy) and femur bone mineral density (BMD) was studied in Wistar rats. After 8 weeks of treatment with 0.5 and 1.0 g kg(-1) day(-1) l-methionine the mean plasma levels of Hcy were 7.67 +/- 1.25 and 61.2 +/- 11.4 micromol/l, respectively. Only rats treated with the higher dose had Hcy levels significantly higher than those of controls, 6.38 +/- 0.90 micromol/l (p < 0.001). Dual Energy X-ray Absorptiometry was used to measure the BMD, which was significantly lower only in the animals with the highest plasma levels of Hcy (p < 0.001). This led us to conclude that increased levels of Hcy are associated with risk of decreased BMD. PMID- 17916930 TI - Effect of long-term fluoride exposure on lipid peroxidation and histology of testes in first- and second-generation rats. AB - This experiment was designed to investigate the histological and lipid peroxidation effects of chronic fluorosis on testes tissues of first- and second generation rats. Sixteen virgin female Wistar rats were mated with eight males (2:1) for approximately 12 h to obtain first-generation rats. Pregnant rats were divided into two groups: controls and fluoride-given group, each of which containing five rats. Pregnant rats in the fluoride-given group were exposed to a total dose of 30 mg/l sodium fluoride (NaF) in commercial drinking water containing 0.07 mg/l of NaF throughout the gestation and lactation periods. After the lactation period, the young animals (first generation, F1) were exposed to the same dose of NaF in drinking water for 4 months. At the end of the 4 months of experimental period, nine randomly chosen male rats (F1) were killed and testes tissues were taken for histopathological and biochemical analysis. The remaining eight female rats were mated with four males (2:1) for approximately 12 h to obtain second-generation rats. Six female were identified as pregnant and treated with similarly throughout the gestation and the lactation periods. After the lactation period, the young male animals (second generation, F2) were also treated in the same way for 4 months. At the end of the 4 months of experimental period, nine randomly chosen male rats (F2) were killed and testes tissues were collected for histopathological and biochemical analysis. The rats in the control group were applied the same procedure without NaF administration. In biochemical analysis of the fluoride given F1 and F2 rats, it has been found that plasma fluoride levels and testes thiobarbituric acid reactive substance levels were significantly increased when compared with the control group. In F1 and F2 rats, similar histopathological changes were observed. In both groups, spermatogenesis was severely reduced. Spermatogonia and primary spermatocytes were normal, however, there was a widespread degeneration in other spermatogenic cell lines of the seminiferous epithelium. The histological structures of the Sertoli and interstitial Leydig cells were normally observed. It is concluded that chronic fluorosis exposure leads to a remarkable destruction in testes tissues of F1 and F2 rats via lipid peroxidation. PMID- 17916931 TI - Mimosine mitigates oxidative stress in selenium deficient seedlings of Vigna radiata. Part II: mitochondrial uptake of 75selenium and mimosine. AB - During the growth of selenium (Se)-deficient seedlings of Vigna radiata, exposure to mimosine [2-amino-3-(3-hydroxy-4-oxo-1H-pyridin-1-yl)-propanoic acid], a nonprotein plant amino acid, effectively mitigated stress at 0.1 mM, as reflected in enhancement of growth and efficiency of mitochondrial functions. Since the changes in the seedlings elicited by exposure to mimosine were similar to those effected by Se at an optimal exposure level of 0.75 ppm (Sreekala et al., Biol Trace Elem Res 70:193-207, 1999), the uptake of Se and that of mimosine itself was individually studied in the respiring mitochondria of Se-deficient seedlings (-Se-stressed group) in comparison with those exposed to mimosine during growth at 0.1 mM (Mim 0.1 group). In both groups, the mitochondrial uptake of (75)Se at 10 microM added Na(2)(75)SeO(3), increased linearly up to 2 min, attaining steady state levels thereafter. Uptake levels were 2.3-fold higher in the Mim 0.1 group than in the -Se-stressed group. Double-reciprocal plots of mitochondrial (75)Se uptake against 2-20 microM Na(2)(75)SeO(3) in the medium were nonlinear and negative cooperative effects during the uptake were confirmed by Scatchard plots, whereas Hill coefficients were 0.8 and 0.85 for the two groups. Mitochondrial uptake of mimosine, at added levels of 25 or 50 microM, increased linearly up to 1 min and decelerated thereafter. Initial uptake levels of mimosine at 1 min were higher by 6.5-fold at 25 microM and 4-fold at 50 microM in the Mim 0.1 group than those in the -Se-stressed group. Initial uptake levels with added mimosine up to 50 or 100 microM yielded nonlinear double-reciprocal plots; and kinetic analyses at 5 to 50 microM revealed the prevalence of positive cooperativity in the -Se stressed group and negative cooperativity in the Mim 0.1 group. Involvement of active thiol groups in the uptake of both Se and mimosine were indicated by inhibition studies. Evidence presented for mimosine mediated increase in mitochondrial Se uptake and cooperative interactions thereof underscores the metabolic significance of mimosine. PMID- 17916932 TI - Effect of magnesium supplementation on some plasma elements in athletes at rest and exhaustion. AB - The effects of magnesium supplementation on plasma magnesium, zinc, and copper levels were determined in young adult tae-kwon-do athletes and sedentary controls at rest and exhaustion. After a 4-week supplementation period with 10 mg/day/kg Mg, the plasma magnesium, copper, and zinc levels significantly increased in sedentary and training (90-120 min training 5 days a week) subjects when compared to nonsupplemented controls (p < 0.05). PMID- 17916933 TI - Moderate accumulation of calcium and phosphorus in the splenic artery with aging and low accumulation of those in the pulmonary artery with aging. AB - To elucidate compositional changes of the arteries with aging, the authors investigated age-related changes of elements in the splenic and pulmonary arteries, which supplied blood to contractile organs. After ordinary dissection by medical students at Nara Medical University was finished, the splenic and pulmonary arteries were resected from the subjects, ranging in age from 58 to 94 years. The element contents were determined by inductively coupled plasma-atomic emission spectrometry. It was found that a moderate accumulation of Ca and P occurred in the splenic artery with aging, but it hardly occurred in the pulmonary artery with aging. Regarding the relationship among elements, the finding that there were significant direct correlations among the contents of Ca, P, Mg, and Na was commonly obtained in both the splenic and pulmonary arteries. The accumulation of Ca and P in the splenic artery with aging occurred independently of that in the pulmonary artery. Histologic observation indicated that a major part of Ca deposits was seen in the middle tunica, but not in the internal tunica. Therefore, the calcification occurring in the splenic artery belonged to middle sclerosis. PMID- 17916934 TI - Increases of calcium, phosphorus, and magnesium in both the right and left fibrous trigones of human heart with aging. AB - To elucidate compositional changes of the cardiac skeleton with aging, the authors investigated age-related changes of elements in both the right and left fibrous trigones of human heart by inductively coupled plasma atomic emission spectrometry (ICP-AES). After ordinary dissection by medical students was finished, hearts were resected from the subjects and both the right and left fibrous trigones were removed from the hearts. The subjects consisted of 10 men and 13 women, ranging in age from 62 to 99 years. The element content was determined by ICP-AES. The Ca, P, and Mg content began to increase in the seventies in both the right and left fibrous trigones, and increased markedly in the eighties. Regarding the relationships among element contents, extremely significant direct correlations were found among the contents of Ca, P, and Mg in both the right and left fibrous trigones. PMID- 17916935 TI - High accumulation of calcium and phosphorus in the pineal bodies with aging. AB - To elucidate compositional changes of the pineal body with aging, the authors investigated age-related changes of elements in the pineal body. After the ordinary dissection by medical students was finished, the pineal bodies and seven arteries were resected from the subjects ranging in age from 58 to 94 years. The element contents were determined by inductively coupled plasma atomic emission spectrometry. It was found that a high accumulation of Ca and P occurred in the pineal bodies with aging. Regarding the relationships among the elements, it was found that there were significant direct correlations among the contents of Ca, P, and Mg. With regard to the relationships between the pineal body and the arteries, no significant correlations were found in the Ca content between the pineal body and the arteries, such as the thoracic and abdominal aortas and the coronary, common carotid, pulmonary, splenic, and common iliac arteries. PMID- 17916936 TI - Changes of porcine growth hormone and pituitary nitrogen monoxide production as a response to cadmium toxicity. AB - The present study was designed to investigate the effects of various cadmium concentrations on porcine growth hormone (GH) secretion in serum and cultured pituitary cells and to explore the possible mechanisms of cadmium toxicity. In feeding trial, 192 barrows (Duroc x Landrace x Yorkshire), with similar initial body weights, were randomly divided into four different treatment groups with three replicates for each treatment. The diets were supplemented for 83 days with 0, 0.5, 5.0, and 10.0 mg/kg cadmium (as CdCl2). For the cell culture trial, dispersed pituitary cells were incubated with graded doses of cadmium (0, 5, 10, 15, or 20 microM) for 24 h. Pigs treated with 10 mg/kg cadmium had significantly decreased serum GH content. 3-(4,5-dimethyl-2-yl)-2,5-diphenyl tetrazolium bromide assay showed that Cd toxicity was dose-dependent. Cell viability was reduced to 50% at 15 microM concentration. Administration of cadmium significantly reduced GH secretion, whereas cellular NO content and inducible nitric oxide synthase activity increased to a certain extent. These findings suggest that the decrease of GH might be related to NO production and to a change of NO signal pathway caused by cadmium. PMID- 17916937 TI - Ultrastructural changes in the kidney of rats with acute exposure to cadmium and effects of exogenous metallothionein. AB - Ultrastructural changes in the kidneys of rats after acute cadmium exposure and the effects of exogenous metallothionein (MT) were studied by transmission electron microscopy. Thirty-six adult Wistar rats were divided into three groups. Cadmium chloride (CdCl2) (3.5 mg/kg/day) was injected subcutaneously in the first group. In the second group, 30 micromol/kg MT was administered in addition to CdCl2. Control rats received 0.5 ml subcutaneous saline solution. Four rats from each group were killed on days 1, 3, 5, and 7 after administration of the compounds. Kidney tissues were taken and fixed in 2.5% glutaraldehyde solution for electron microscopic observations. Tissue damage in kidney increased as time passed since the administration of CdCl2 in the first group. Degeneration in the proximal and distal tubules was observed. Increased apoptosis was seen in the proximal tubules epithelium, especially on day 7. Peritubular capillaries became dilated, there was degeneration of the endothelial cells, and the amount of intertubular collagen fibers was increased. On day 1, irregular microvilli in the proximal tubules, deepening of the basal striations, and myelin figures; on day 3, multiple vesicular mitochondria and regions of edema around tubules; on days 5 and 7, increased apoptotic cell in the proximal tubules and widened rough endoplasmic reticulum of the endothelial cells of glomerular capillaries were observed. We observed that the structural alterations that increased depending on the day of Cd administration decreased after exogenous MT administration, the dilation of the peritubular capillaries persisted, and there were degenerated proximal tubules. It was established that cadmium chloride was toxic for kidney cortex and caused structural damage. Exogenous MT partly prevents CdCl2-induced damage. PMID- 17916938 TI - Anticarcinogenic effects of hexaamminecobalt(III) chloride in mice initiated with diethylnitrosamine. AB - Hexaamminecobalt(III) chloride ([Co(NH3)6]Cl3) was investigated for its antineoplastic role in relation to tumor marker enzymes, drug metabolizing enzymes, oxidative stress-related parameters, and histopathological analysis of liver and lung tissues of mice. Initiation was performed using a single intraperitoneal injection of diethylnitrosamine (DENA) at a carcinogenic dose of 90 mg/kg body weight. The cobalt complex supplementation at a dose of 100 ppm in drinking water was given ad libitum throughout the experimental period of 14 weeks. In comparison to lung, the cobalt complex supplementation was found to reverse DENA-induced biochemical changes more effectively in liver. Histological examination of liver and lung from DENA-initiated and cobalt-complex-supplemented mice showed considerable protection in the case of liver compared to that of lung. The involvement of the [Co(NH3)6]Cl3 in modulating several factors associated with carcinogenesis induced by DENA thus showed its anticarcinogenic potential against chemically induced hepatocarcinogenesis. PMID- 17916939 TI - Zinc distribution and expression pattern of ZnT3 in mouse brain. AB - To explore the relationship between the zinc distribution and zinc transporter 3 (ZnT3) mRNA expression in the mouse brain, zinc contents and its distribution were determined by synchrotron radiation x-ray fluorescence (SRXRF), and ZnT3 mRNA expression was examined by reverse-transcription polymerase chain reaction and in situ hybridization. The results showed that the zinc contents were not distributed evenly in various brain tissues. The zinc contents in cerebral cortex and hippocampus were nearly 5-10 times higher than that in other neural locations. Correspondingly, ZnT3 mRNA expression was observed in high abundance in the cerebral cortex, hippocampus, and testis, but was not detected in other organs and tissues. In the nervous system, ZnT3 mRNA was detected mainly in hippocampus, cerebral cortex, and spinal ganglion. The present results show the coincident distribution of zinc and ZnT3 mRNA in mouse brain. The high zinc contents might be determined by the high expression of ZnT3. More meaningfully, the results showed the feasibility of applying of SRXRF in examining the distribution of minerals in different organs and tissues. In addition, it was observed for the first time that ZnT3 mRNA was expressed in the facial nucleus. The function of ZnT3 in facial nucleus awaits further study. PMID- 17916940 TI - Microcalorimetric study on the toxic effect of Pb2+ to Tetrahymena. AB - The toxic effect of Pb2+ has been studied in eukaryotic cells by using Tetrahymena as a target. The maximum power (P(m)) and the growth rate constant (k) were determined, which showed that values of P(m) and k were linked to the concentration (C) of Pb2+. The addition of Pb2+ caused a decrease of the maximum heat production and growth rate constant, indicating that Tetrahymena growth was inhibited in the presence of Pb2+, and Pb2+ took part in the metabolism of cells. From micrographs, morphological changes of Tetrahymena were observed with addition of Pb2+, indicating that the toxic effect of Pb2+ derived from destroying the membrane of surface of Tetrahymena. According to the thermogenic curves and photos of Tetrahymena under different conditions, it is clear that metabolic mechanism of Halobacterium halobium R1 growth has been changed with the addition of Pb2+. PMID- 17916941 TI - Promotion of energy transfer and oxygen evolution in spinach photosystem II by nano-anatase TiO2. AB - Being a proven photocatalyst, nano-anatase is capable of undergoing electron transfer reactions under light. In previous studies we had proven that nano anatase improved photosynthesis and greatly promoted spinach growth. The mechanisms by which nano-anatase promotes energy transfer and the conversion efficiency of the process are still not clearly understood. In the present paper, we report the results obtained with the photosystem II (PSII) isolated from spinach and treated by nano-anatase TiO2 and studied the effect of nano-anatase TiO2 on energy transfer in PSII by spectroscopy and on oxygen evolution. The results showed that nano-anatase TiO2 treatment at a suitable concentration could significantly change PSII microenvironment and increase absorbance for visible light, improve energy transfer among amino acids within PSII protein complex, and accelerate energy transport from tyrosine residue to chlorophyll a. The photochemical activity of PSII (fluorescence quantum yield) and its oxygen evolving rate were enhanced by nano-anatase TiO2. This is viewed as evidence that nano-anatase TiO2 can promote energy transfer and oxygen evolution in PSII of spinach. PMID- 17916942 TI - Proceedings of the international symposium on selenium-mercury interactions. PMID- 17916943 TI - Mercury-selenium species ratio in representative fish samples and their bioaccessibility by an in vitro digestion method. AB - The potential toxicity of mercury (Hg) content in fish has been widely evaluated by the scientific community, with Methylmercury (MeHg) being the only legislated species (1 mg kg-1, maximum concentration allowed in predatory fish). On the other hand, selenium (Se) is recognized to decrease its toxicity when both elements are simultaneously administrated. In the present paper, the total content of Se and Hg and their species in fish of high consumption, such as tuna, swordfish, and sardine, have been evaluated. The percentage of MeHg is higher than 90% of total Hg content. The results show that, for all of them, the Se/Hg ratio is significantly higher than one, being the maximum ratio for sardine. As only studying the bioaccessible fraction the extent of a toxic effect caused by an element can be predicted, the bioaccessibility of both analytes through an in vitro digestion method has been carried out. The results show that MeHg in all fishes is very low bioaccessible in both gastric and intestinal digestion. Because the MeHg bioaccessible fraction might be correlated to the Se content, the potential toxicity cannot be only related to the total Hg content but also to Se/Hg ratio. PMID- 17916944 TI - Selenium-mercury interactions in man and animals. AB - Selenium-mercury interactions were most extensively studied in relation to alleviation of Hg toxicity by added selenium. This presentation considers the influence of mercury on endogenous selenium, on its tissue and cellular "status" after lifelong or acute exposure to mercury vapor (Hg o). Discussed are data obtained from (1) humans living near or working in a mercury mine, and (2) rats experimentally exposed in the mine. Mercury vapor is unique--or similar to methylmercury--because of its ability to penetrate cell membranes and so invade all cells, where it is oxidized in the biologically active form (Hg++) by catalase. Such in situ-generated ions can react with endogenously generated highly reactive Se metabolites, like HSe-, and render a part of the selenium unavailable for selenoprotein synthesis. Data on human populations indicate that in moderate Hg exposure combined with an adequate selenium supply through diet, Se bioavailability can be preserved. On the other hand, the results of an acute exposure study emphasize the dual role of selenium in mercury detoxification. Besides the well-known Se coaccumulation through formation of nontoxic Hg-Se complexes, we observed noticeable Se (co)excretion, at least at the beginning of exposure. The higher Hg accumulation rate in the group of animals with lower basal selenium levels can also point to selenium involvement in mercury excretion. In such conditions there is a higher probability for decreased selenoprotein levels (synthesis) in some tissues or organs, depending on the synthesis hierarchy. PMID- 17916945 TI - Factors in fish modifying methylmercury toxicity and metabolism. AB - We report here some results of a long-term (19 month) study with cats fed methylmercury (MeHg) in nutritionally balanced diets based on fish. By using either freshwater pike (low in Se) or canned tuna (high in Se) as the major protein source, basal diets with low levels of MeHg were prepared having different Se content, all Se being of natural origin. The basal diets produced no signs of toxicity or pathological changes over the l9-month period. In cats fed basal diets spiked with medium or high levels of MeHg, evidence for delayed onset of toxic effects from the added MeHg was observed with the tuna diets compared to pike diets. In brain, muscle, and blood, the activity of GSH peroxidase, a selenoenzyme, was decreased by Hg. In liver, substantial accumulation of Hg with Se occured (molar Hg/Se ratio approximately 1.4 to 1.8) but GSH peroxidase activity was unaffected. We suggest that the coaccumulation of Hg and Se in liver measures the extent to which MeHg has been metabolically transformed by metabolism to Hg++, and inactivated by deposition as a Hg/Se complex of low bioavailability. The accumulation of Hg and Se in liver was much greater in cats fed tuna compared to pike, out of proportion to the relatively small differences in Hg and Se content of the tuna and pike basal diets. Some mechanisms are described by which selenium, vitamin E, and other factors might facilitate MeHg breakdown to inorganic Hg during long term low level exposure to MeHg. PMID- 17916946 TI - New developments in selenium biochemistry: selenocysteine biosynthesis in eukaryotes and archaea. AB - We used comparative genomics and experimental analyses to show that (1) eukaryotes and archaea, which possess the selenocysteine (Sec) protein insertion machinery contain an enzyme, O-phosphoseryl-transfer RNA (tRNA) [Ser]Sec kinase (designated PSTK), which phosphorylates seryl-tRNA [Ser]Sec to form O phosphoseryl-tRNA [Ser]Sec and (2) the Sec synthase (SecS) in mammals is a pyridoxal phosphate-containing protein previously described as the soluble liver antigen (SLA). SecS uses the product of PSTK, O-phosphoseryl-tRNA[Ser]Sec, and selenophosphate as substrates to generate selenocysteyl-tRNA [Ser]Sec. Sec could be synthesized on tRNA [Ser]Sec from selenide, adenosine triphosphate (ATP), and serine using tRNA[Ser]Sec, seryl-tRNA synthetase, PSTK, selenophosphate synthetase, and SecS. The enzyme that synthesizes monoselenophosphate is a previously identified selenoprotein, selenophosphate synthetase 2 (SPS2), whereas the previously identified mammalian selenophosphate synthetase 1 did not serve this function. Monoselenophosphate also served directly in the reaction replacing ATP, selenide, and SPS2, demonstrating that this compound was the active selenium donor. Conservation of the overall pathway of Sec biosynthesis suggests that this pathway is also active in other eukaryotes and archaea that contain selenoproteins. PMID- 17916947 TI - Selenium and mercury in pelagic fish in the central north pacific near Hawaii. AB - Protective effects of selenium against mercury toxicity have been demonstrated in all animal models evaluated. As interactions between selenium and mercury and their molar ratios in seafood are essential factors in evaluating risks associated with dietary mercury exposure, considering mercury content alone is inadequate. In this study, the absolute and molar concentrations of mercury and selenium were determined in edible portions from 420 individual fish representing 15 species of pelagic fish collected from the central North Pacific Ocean near Hawaii. Selenium was in molar excess of mercury in almost all fish species evaluated. The rank order of mean Se/Hg molar ratios was striped marlin (17.6) > yellowfin tuna (14.1) > mahimahi (13.1) > skipjack tuna (12.8) > spearfish (11.4) > wahoo (10.8) > sickle pomfret (6.7) > albacore tuna (5.3) > bigeye tuna (5.2) > blue marlin (4.1) > escolar (2.4) > opah (2.3) > thresher shark (1.5) > swordfish (1.2) > mako shark (0.5). With a Se/Hg molar ratio of less than 1, mako shark was the only fish containing a net molar excess of mercury. A selenium health benefit value based on the absolute amounts and relative proportions of selenium and mercury in seafood is proposed as a more comprehensive seafood safety criterion. PMID- 17916948 TI - Importance of molar ratios in selenium-dependent protection against methylmercury toxicity. AB - The influence of dietary selenium (Se) on mercury (Hg) toxicity was studied in weanling male Long Evans rats. Rats were fed AIN-93G-based low-Se torula yeast diets or diets augmented with sodium selenite to attain adequate- or rich-Se levels (0.1, 1.0 or 15 micromol/kg, respectively) These diets were prepared with no added methylmercury (MeHg) or with moderate- or high-MeHg (0.2, 10 or 60 micromol/kg, respectively). Health and weights were monitored weekly. By the end of the 9-week study, MeHg toxicity had impaired growth of rats fed high-MeHg, low Se diets by approximately 24% (p < 0.05) compared to the controls. Growth of rats fed high-MeHg, adequate-Se diets was impaired by approximately 8% (p < 0.05) relative to their control group, but rats fed high-MeHg, rich-Se diets did not show any growth impairment. Low-MeHg exposure did not affect rat growth at any dietary Se level. Concentrations of Hg in hair and blood reflected dietary MeHg exposure, but Hg toxicity was more directly related to the Hg to Se ratios. Results support the hypothesis that Hg-dependent sequestration of Se is a primary mechanism of Hg toxicity. Therefore, Hg to Se molar ratios provide a more reliable and comprehensive criteria for evaluating risks associated with MeHg exposure. PMID- 17916949 TI - The impact of high-dose sodium selenite therapy on Bcl-2 expression in adult non Hodgkin's lymphoma patients: correlation with response and survival. AB - The present study was undertaken to explore the effect of administration of high doses of sodium selenite on the expression of Bcl-2 in patients with non Hodgkin's lymphoma (NHL). Fifty patients with newly diagnosed NHL were randomly divided into two groups. Group A-I received standard chemotherapy whereas group A II received adjuvant sodium selenite 0.2 mg kg-1 day-1 for 30 days in addition to chemotherapy. Enzyme-linked immunosorbent assay was used to assess Bcl-2 at the time of diagnosis and after therapy in the two groups. Sodium selenite administration resulted in significant decline of Bcl-2 level after therapy in group A-II (8.6 +/- 6.9 ng/ml vs 3 6.9 +/- 7.9 ng/ml, P < 0.05). Also, complete response reached 60% in group A-II compared to 40% in group A-I. Significant increase in CD4/CD8 ratio was noticed in group A-II compared to group A-I after therapy (1.45 +/- 0.36 vs 1.10 +/- 0.28 p 0.04). Overall survival time in months was significantly longer in complete remission patients in group A-II (21.87 +/- 1.41) compared to group A-I (19.70 +/- 1.95) (p = 0.01). It is concluded that sodium selenite administration at the dosage and duration chosen acts as a down regulator of Bcl-2 and improves clinical outcome. PMID- 17916950 TI - Zinc nutritional status in adolescents with Down syndrome. AB - Studies have evidenced that zinc metabolism is altered in presence of Down syndrome, and zinc seems to have a relationship with the metabolic alterations usually present in this syndrome. In this work, the Zn-related nutritional status of adolescents with Down syndrome was evaluated by means of biochemical parameters and diet. A case-control study was performed in a group of adolescents with Down syndrome (n = 30) and a control group (n = 32), of both sexes, aged 10 to 19 years. Diet evaluation was accomplished by using a 3-day dietary record, and the analysis was performed by the NutWin program, version 1.5. Antropometric measurements were performed for evaluation of body composition. The Zn-related nutritional status of the groups was evaluated by means of zinc concentration determinations in plasma and erythrocytes, and 24-h urinary zinc excretion, by using the method of atomic absorption spectroscopy. The diet of both groups presented adequate concentrations of lipids, proteins, carbohydrates, and zinc. The mean values found for zinc concentration in erythrocytes were 49.2 +/- 8.5 microg Zn/g Hb for the Down syndrome group and 35.9 +/- 6.1 microg Zn/g Hb for the control group (p = 0.001). The average values found for zinc concentration in plasma were 67.6 +/- 25.6 microg/dL for the Down syndrome group and 68.9 +/- 22.3 microg/dL for the control group. The mean values found for zinc concentration in urine were 244.3 +/- 194.9 microg Zn/24 h for the Down syndrome group and 200.3 +/- 236.4 microg Zn/24 h for the control group. Assessment of body composition revealed overweight (26.7%) and obesity (6.6%) in the Down syndrome group. In this study, patients with Down syndrome presented altered zinc levels for some cellular compartments, and the average zinc concentrations were low in plasma and urine and elevated in erythrocytes. PMID- 17916951 TI - Increased plasma manganese, partially reduced ascorbate, 1 and absence of mitochondrial oxidative stress in type 2 diabetes mellitus: implications for the superoxide uncoupling protein 2 (UCP-2) pathway. AB - Oxidative stress is an important component of diabetes and its complications. Manganese (Mn), the key component of the Mitochondrial antioxidant (MnSOD), plays a key role in the superoxide uncoupling protein 2 (UCP-2) pathway in inhibiting of glucose-stimulated insulin secretion (GSIS). The interactions of Mn with ascorbate and other components of this pathway have not been defined in type-2 diabetes. Fifty established type 2 diabetics (30 males, 20 females) and 30 non diabetics (controls; 18 males, 12 females) matched for age and sex were investigated. Dietary intake, particularly of micronutrients as assessed by 24-h dietary recall was similar between diabetics and controls. Weight and height of all subjects were determined and body mass index (BMI) computed after clinical assessment. Fasting plasma glucose, manganese, ascorbic acid, creatinine and K+ levels were determined; K+ was to assess the K+ channels, whereas creatinine was to assess probability of oxidative stress nephropathy. Body mass index was greater in DM than in controls (p < 0.001). Fasting plasma glucose and Mn levels (p < 0.00 and p < 0.01, respectively) were higher in diabetes than in the controls. Manganese level was greater than twice the levels in controls. Ascorbic acid was not significantly different (p > 0.05), but was 50% lower than the level in non-diabetics. Potassium like Mn and glucose was significantly higher in diabetes mellitus (DM) than in controls (p < 0.001). Creatinine was not significantly different between diabetics and controls (p > 0.05). Correlations among all parameters were not significantly different. These findings suggest absence of significant oxidative stress in the mitochondria, probably excluding a role for UCP-2-superoxide pathway in the inhibition of glucose-stimulated insulin secretion (GSIS), calling for caution in the precocious conclusion that interruption of UCP-2 activity may provide a viable strategy to improve beta-cell dysfunction in type 2 diabetes mellitus. PMID- 17916952 TI - Clinical evaluation of the bioavailability of zinc-enriched yeast and zinc gluconate in healthy volunteers. AB - Zinc (Zn)-enriched yeast and gluconate are considered two of the more biologically available supplements. However, there have been few reports comparing the bioavailability of these supplements. The objective of this study was to demonstrate whether Zn was absorbed better by healthy male volunteers when given supplements where the mineral is found organically bound in yeast or as a salt gluconate form. The trial used a randomized, two-way crossover design. Urine, blood, and fecal samples were collected and analyzed over a 48-h period after a single dose of supplement. The net Zn balance and the relative bioavailability were calculated. No differences were observed in urine excretion of the two supplements. Zinc gluconate gave higher Zn concentrations in the blood in the first 6 h but also showed greater losses in the feces. Zinc yeast also increased in blood with time but showed significantly less loss in the feces. Thus, the net Zn balance after 48 h for Zn yeast was 9.46 but for Zn gluconate it was -2.00, indicating that Zn gluconate supplementation contributed to a net loss of Zn. It was concluded that organic Zn yeast supplements are more biologically available than Zn gluconate salts. PMID- 17916953 TI - Urinary profile of 6 beta-hydroxycortisol in workers exposed to aluminum. AB - The aim of the present study was to evaluate whether aluminum exposure may cause alterations of cytochrome P4503A4 by measuring the urinary excretion of 6 beta hydroxycortisol as biomarker. There is a positive correlation between urinary 6 beta-hydroxycortisol and aluminum excretion (p < 0.05). The correlations between occupational duration, smoking habits, and age were also evaluated. The increased excretion of the biomarker suggests that this element may induce formation of cytochrome P 450 in exposed individuals. PMID- 17916954 TI - Urinary excretion of zinc and metabolic control of patients with diabetes type 2. AB - The objective of this study was to assess urinary excretion of zinc and evaluation parameters of metabolic control in type 2 diabetic patients. Thirty one type 2 diabetic patients, of both genders, with 5.8 +/- 5.6 years average time of the disease, age range 20-60 years, were selected. Evaluation of the nutritional status was performed using anthropometric measurements. To evaluate food consumption, the 3-day alimentary log method was used, and its analysis was performed using a software. Determination of urinary zinc was by atomic absorption spectrophotometry. From the obtained results, it was concluded that 51.6% of the patients were overweight. The mean of found waist circumference was 100.4 and 92.2 cm for men and women, respectively. Blood glucose and glycated hemoglobin values were higher than reference values, and plasma albumin concentration was adequate. The median of found urinary zinc excretion was 474.9 mug/24 h, within normal standards (300-600 mug/day). Regarding diet composition, calorie and protein concentration were above recommendation, while mean zinc concentration was adequate. This data allow the conclusion that the evaluated patients presented adequate urinary zinc excretion in comparison with reference values. PMID- 17916955 TI - The interaction between selenium status, sex hormones, and thyroid metabolism in adolescent girls in the luteal phase of their menstrual cycle. AB - The objective of the present work was to study all physiological relationships among selenium status (SeS), sex hormones secretion (SH), and thyroid metabolism (ThM) in healthy adolescent girls, at one time. Forty-four girls aged 13.4-16.6 years (mean age, 14.5 +/- 0.5 years) entered the statistical model. Parameters reflecting SeS: plasma selenium concentration (Se) and plasma glutathione peroxidase activity (GPX3); SH: serum estradiol (E2) and progesterone (P4); age of menarche (AoM); and ThM: thyroid stimulating hormone (TSH), free thyroxine (fT4), free triiodothyronine (fT3), antithyroid peroxidase antibodies (anti-TPO) in serum, and thyroid volume (ThV), were determined, and the interactions between them were evaluated by means of the partial least squares method (PLS). PLS method was, for the first time, successfully applied to the problem of selenium and hormone interactions and revealed that selenium status and female reproductive system are interrelated and affect thyroid physiology in adolescent girls in the luteal phase. The strongest associations were revealed for the pairs of parameters, Se and fT4/fT3, Se and P4, the modest ones for the pairs, Se and ThV, P4 and fT4/fT3, Se and AoM, and P4 and AoM. There was no correlation between E2, GPX3, and TSH, and any other considered parameter. Se and P4 had the greatest influence on ThM parameters. PMID- 17916956 TI - Mineral factors controlling essential hypertension--a study in the Chandigarh, India population. AB - Essential hypertension (EH) is a major public health problem world over and in India. Recent data on EH in the population of Chandigarh (Union Territory and capital of Punjab and Haryana States of India) revealed that the prevalence of EH has become double in the last 30 years in the residents of Chandigarh (26.9 to 45.80% in the year 1968 and 2002). Zinc (Zn), copper (Cu), magnesium (Mg), and manganese (Mn) in the serum are considered important in maintaining the human hypertension. The high Zn intake was considered to increase the blood pressure (BP) and to affect the other mineral status in the body. Recent survey on the trace metal status of different vegetables in the State of Punjab around Chandigarh (India) revealed that Zn level is significantly higher (40 mg/kg or more in above ground vegetables and 120 mg/kg or above in underground vegetables) in underground water-irrigated vegetables, but the levels of Cu and Mg are within prescribed limit. The present study was conducted on Chandigarh population to evaluate the levels of Zn, Cu, Mg, and Mn in the blood and urine of normotensive (NT) control and hypertensive (HT) subjects matched with number, age and sex. Atomic absorption spectrophotometer studies reevaluated that the levels of serum Zn, Mg, and Mn were significantly higher (p < 0.001), but the level of Cu was low in the HT subjects (BP = 160/93) compared to NT control (BP = 140/83). Higher levels of urinary Zn, Cu, Mg, and Mn were observed in the HT subject vs NT control (p < 0.001). Positive correlations were evaluated between the levels of serum Zn, Mg, and Mn vs systolic and diastolic pressures (DP and SP), respectively (r = 0.928, 0.863, 0.876, 0.808, 0.404, 0.326, p < 0.01), but negative and positive nonsignificant correlations between the serum Cu with SP and DP were recorded (r = -0.032, r = 0.024). Positive correlations were also evaluated between urinary levels of Zn, Cu, Mg, and Mn vs SP and DP (r = 0.718, 0.657, 0.750, 0.681, 0.630, 0.578, 0.516, 0.461, p < 0.01). Prevalence of essential hypertension may be due to higher Zn level in the food chain that makes the individuals vulnerable to other diseases over the time related to essential hypertension. PMID- 17916957 TI - Differences in the element contents between gunshot entry wounds with full jacketed bullet and lead bullet. AB - To elucidate characteristics of gunshot residues in gunshot entry wounds with full-jacketed and lead bullets, element contents in entry gunshot wounds and control skins were analyzed by inductively coupled plasma-atomic emission spectrometry (ICP-AES). It was found that a high content of Fe and Zn was deposited in the gunshot entry wounds with full-jacketed bullet, whereas a high content of Pb was deposited in the gunshot entry wounds with lead (unjacked) bullet. It should be noted that the content of Pb was significantly higher in the gunshot entry wounds with lead bullet than in those with full-jacketed bullet. Regarding the relationships among elements, it was found that there were significant direct correlations between Pb and either Sb or Ba contents in both gunshot entry wounds with full-jacketed and lead bullets. As Pb increased in both gunshot entry wounds, Sb and Ba also increased in the wounds. PMID- 17916958 TI - High levels of cadmium and lead in seminal fluid and blood of smoking men are associated with high oxidative stress and damage in infertile subjects. AB - We measured the levels of malondialdehyde (MDA), protein carbonyls, glutathione S transferase (GST) and reducte glutathione (GSH) in seminal plasma and spermatozoa from 95 subjects including 50 infertile patients to evaluate the association between oxidative stress and damage and the components of the anti-oxidant defenses in seminal plasma and spermatozoa of infertile subjects and concentrations of cadmium (Cd) and lead (Pb) in the blood and seminal plasma because of tobacco smoke exposure. The reactive oxygen species (ROS) in spermatozoa were also evaluated by luminol (5-amino-2,3-dihydro-1,4 phthalazinedione)-enhanced chemiluminescence assay. The sperm count, motility, and morphology in the smokers infertile group were found to be lower than those in the fertile male group and nonsmokers infertile group (p < 0.001). Concentrations of Cd, Pb, MDA, protein carbonyls, and ROS levels in the smokers infertile group were significantly higher than those in the fertile male and nonsmokers infertile male groups (p < 0.001). However, GSH levels and GST activities were decreased in the smokers infertile male group than those in the fertile male and nonsmokers infertile male groups (p < 0.001). The results indicate that smoking could affect semen quality and oxidative lipid and protein damage in human spermatozoa. From Pearson correlation analysis, positive correlations were demonstrated between the seminal plasma Cd and seminal plasma protein carbonyls and between seminal plasma Pb and spermatozoa ROS levels in smokers of the subfertile group, while there was a significant positive correlation between blood Cd and ROS levels in smokers of the fertile group. There was also a significant negative correlation of the Cd level of the blood and GSH levels of the sperm and seminal plasma. These findings suggest that cigarette smoking enhances the levels of Cd and Pb in seminal plasma and blood and the extent of oxidative damage associated with a decrease in components of the anti-oxidant defenses in the sperm of infertile males. PMID- 17916959 TI - Acquisition of visuomotor abilities and intellectual quotient in children aged 4 10 years: relationship with micronutrient nutritional status. AB - Lethargy, poor attention, and the high rate and severity of infections in malnourished children affect their educational achievement. We therefore studied the association between visuomotor abilities and intelligence quotient (IQ) and their relationship with iron, zinc, and copper. A cross-sectional study was carried out on a sample of 89 healthy children (age range, 4-10 years). Evaluations of visuomotor ability and IQ were performed with the Developmental Test of Visual Motor Integration (VMI) and the Scale for Measurement of Intelligence for children aged 3-18 years, respectively. Nutritional status was assessed using anthropometry and biochemical assessments, which included serum ferritin, zinc and copper levels, and Hb. The sample was classified as having low or normal VMI scores: 47 children (52.8%, mean age 7 +/- 1.5 years) had low VMI, and 42 (47.2%, mean age 7 +/- 2.06 years) had normal VMI. There were no statistically significant differences in socioeconomic and cultural condition between both groups. We found significantly higher serum copper and ferritin levels in normal as compared to low VMI, but we did not find any differences with zinc. IQ was significantly higher in normal vs low VMI children. The fact that children with abnormal VMI presented low mean serum copper and ferritin concentrations could indicate that copper and iron deficiencies in this sample could be related with visuomotor abilities. PMID- 17916960 TI - Effect of various clinical variables on total intracellular magnesium in hospitalized normomagnesemic diabetic patients before discharge. AB - Deficiency of intracellular magnesium (icMg) may coexist with normal serum Mg levels. Little is known about clinical and pharmacological factors affecting icMg in normomagnesemic patients with diabetes mellitus (DM). Moreover, no information exists regarding the icMg state in diabetic patients after acute illness and before hospital discharge. We have evaluated the effect of antihyperglycemic medications and other relevant clinical variables on icMg in 119 such patients. Total icMg was measured in peripheral blood mononuclear cells. Twenty healthy volunteers served as controls. IcMg content (microg/mg cell protein) was lower in DM compared to controls (1.74 +/- 0.44 vs 2.4 +/- 0.39, p < 0.001). It was also significantly lower in patients treated with insulin (1.57 +/- 0.31 vs 1.8 +/- 0.46, p = 0.01), while metformin treatment was associated with higher icMg (1.86 +/- 0.49 vs 1.63 +/- 0.35, p = 0.003). After adjustment for age, gender, and concomitant use of other hypoglycemic drugs, only treatment with metformin was independently associated with increased icMg (p = 0.03). No statistically significant association or correlation was found between icMg content and age, causes of hospitalization, comorbid conditions, treatment with other drugs, concentrations of HbA1c, serum glucose, Mg, or creatinine. In conclusion, icMg is depleted in normomagnesemic DM patients. Insulin treatment is associated with worsening of icMg status, while metformin treatment may confer protective effect. PMID- 17916961 TI - Bioavailability studies of stabilized iron (II) sulfate by means of the prophylactic-preventive method. AB - The bioavailability of stabilized ferrous sulfate was studied by means of the prophylactic-preventive test in rats. For comparative purposes, ferrous sulfate was used as reference standard. The test was performed in male weaned rats during 3 weeks, which were randomized into three groups of ten animals each. A control group received a basal diet of low iron content while the other groups received the same diet added with iron at a dose of 15 mg/kg as FeSO4 7H2O and stabilized ferrous sulfate, respectively. Individual hemoglobin concentrations and weights were determined at the beginning and at the end of the study, and food intake was daily registered. Iron bioavailability (BioFe) of each source was calculated as the ratio between the amount of iron incorporated into hemoglobin during the treatment and the total iron intake per animal. A relative biological value was obtained as the ratio between the BioFe of stabilized ferrous sulfate and the reference standard given a value of 96%. Stabilized ferrous sulfate showed a high bioavailability, and when it is used to fortify dairy products as cheese and fluid milk in a dose of 15-20 mg of iron per kilogram, it behaved inertly in relation to the sensorial properties of the fortified food. These results suggest that this iron compound is a promising source to be use in food fortification. PMID- 17916962 TI - Fluoride ingestion after brushing with a gel containing a high concentration of fluoride. AB - The aim of the study was to evaluate the amount of fluoride remaining in the oral cavity of children after brushing with fluoride gel (1.25% F). The study involved six groups of 7-year-old and six groups of 11-year-old children. The procedure was carried out according to the manufacturer's recommendations. Fluoride concentrations were determined using ion-selective fluoride electrode. No statistically significant difference was found between the amount of fluorides that remained in the oral cavity of younger and older age group (1.2 and 1.3 mg, respectively; p > 0.05). The amount of fluorides swallowed during the procedure in both age groups proves to be within acceptable limit, as far as risk of acute poisoning symptoms is concerned. The individual daily fluoride exposure during the day of procedure seems to be twice as high compared to average fluoride intake from diet and dentifrice, and it does not exceed Tolerable Upper Intake Level for children more than 8. In younger children, it seems justifiable to reduce the amount of the preparation applied on a toothbrush, especially when daily use of the gel is recommended. PMID- 17916963 TI - Effects of environmental lead pollution, smoking, and smokeless tobacco (Maras powder) use on blood lead level. AB - One hundred sixty-four adult male volunteers (29 controls [Group 1] and 135 combi drivers) enrolled in the study. The combi drivers were divided into three groups as nonusers of either Maras powder or cigarette (Group 2), smokers (Group 3), and users of Maras powder (Group 4). Blood lead levels (BLLs) were analyzed by atomic absorption spectrophotometer. BLL was detected as 2.8 +/- 2.3 microg/dL in Group 1 (n = 29); however, it was 3.5 +/- 1.6 microg/dL in Group 2 (n = 33), 3.8 +/- 2.4 microg/dL in Group 3 (n = 62), and 3.9 +/- 2.4 microg/dL in Group 4 (n = 40). BLL in Group 1 was found significantly lower than other groups (p < 0.05). The use of cigarette or Maras powder by the drivers did not give rise to a marked difference on the BLLs (p > 0.05). BLL of (combi) drivers was detected to be significantly higher than nondrivers; however, it was still under the hazardous level of 10 microg/dL announced by WHO. Although there are publications reporting that usage of tobacco increases the level of lead in blood, both smoking and use of Maras powder did not affect BLL markedly in our study. PMID- 17916964 TI - Disruption of elements uptake due to excess chromium in Indian medicinal plants. AB - Chromium and its compounds may cause disturbance in the nutrient level of the plants. Iron, manganese, copper, and zinc are essential nutrient elements and required for balanced growth and development of plants, but chromium uptake sometimes disturbed their concentration in plants. Therefore, in the present paper, an effort has been made to observe the effect of different levels of Cr on nutrient uptake of Phyllanthus amarus and Solanum nigrum, the medicinally important plants of indigenous systems of medicine having hepatoprotective and diuretic properties. The study revealed that Cr causes significant changes in nutrient uptake as compared to control plants. Besides, Cr-treated plants showed growth depression and decrease in fresh and dry weight too. With the increase in Cr supply, accumulation of Cr in roots was increased significantly. Concentration of manganese and zinc was also increased. However, copper concentration in both the plants seemed less affected by Cr. PMID- 17916965 TI - Effect of zinc upon human and murine cell viability and differentiation. AB - Most zinc studies show its benefits or changes that coincide with its deficiency, but some have reported damages by supplements. In this work, the effects of zinc in different cell lines (U-937, human monocytes, and murine bone marrow cells) were analyzed. The cells were put in their specific culture medium either alone or with a stimulant [1-phorbol 12-myristate 13-acetate (PMA) for U-937 and monocytes, granulocyte macrophage colony stimulating factor (GM-CSF) for bone marrow cells]. These preparations, with or without zinc (0.05 to 1.0 mM), were incubated and microscopically analyzed on days 3, 9, and 11. The viability of all cells cultivated with 0.05 and 0.1 mM of zinc was similar to that of the controls without zinc (90%). With 1.0 mM of zinc, the viability diminished (p < 0.005) to 80% in U-937 and to 50% in monocytes and bone marrow cells; the number of cells increased in the three lines, but there was no differentiation. We conclude that the effects observed with different doses of zinc vary not only among the different species but also according to the time the cells were exposed to the metal. The same doses of zinc can have either a stimulatory or an inhibitory effect. PMID- 17916966 TI - Structural basis for the biological effects of Pr(III) ions: alteration of cell membrane permeability. AB - The biological effects of rare-earth ions on the organism have been studied using Pr3+ as a probe ion and Escherichia coli cell as a target. Atomic force microscopy (AFM) observation of the surface of E. coli cells shows that the presence of Pr3+ substantially changes the structure of the outer membrane. By induced coupled plasma-mass spectrometry (ICP-MS), more Cu2+ was found in the cells grown in the presence of Pr3+, indicating changes of cell permeability. Using energy dispersive X-ray spectroscopy (EDX), Ca2+ is found on the outer surface of the original cell. It is proposed that Pr3+ can replace Ca2+ from the binding sites because of their close ionic radii and similar ligand speciality. PMID- 17916967 TI - Availability of essential elements in nutrient supplements used as antidiabetic herbal formulations. AB - Five brands of antidiabetic herbal formulations as tablets, Diabetex, Divya Madhu Nashini, Jambrushila, Diabeticin, and Madhumeh Nashini, from different pharmacies were analyzed for six minor (Na, K, Ca, Cl, Mg, and P) and 20 trace (As, Ba, Br, Ce, Co, Cr, Cs, Cu, Fe, Hg, La, Mn, Rb, Sb, Sc, Se, Sm, Th, V, and Zn) elements by thermal neutron irradiation followed by high-resolution gamma ray spectrometry. Further Ni, Cd, and Pb were determined by atomic absorption spectrophotometry. Most elements vary in a narrow range by a factor of 2-4 while a few others vary in a wide range, e.g., Na (0.05-0.67 mg/g), Mn (26.7-250 microg/g), and V (0.26-2.50 microg/g). All the five brands contain K, Cl, Mg, P, and Ca as minor constituents along with mean trace amounts of Cr (2.11 +/- 0.67 microg/g), Cu (15.7 +/- 7.11 microg/g), Fe (459 +/- 171 microg/g), Mn (143 +/- 23 microg/g), Se (238 +/- 112 ng/g), and V (0.99 +/- 0.93 microg/g). Jambrushila is enriched in Na, Ca, Mg, Cl, Fe, Cu, Se, and Zn, essential nutrients responsible for curing diabetes. Dietary intake of Mn, Fe, and Cu are greater than 10% of the recommended dietary allowance, whereas that for Zn and Se is less than 2%. Mean contents of toxic elements (As, Cd, Hg, and Pb) were found below permissible limits except in Jambrushila. Cr and Zn were inversely correlated with r = -0.81, whereas Rb and Cs exhibit linear correlation (r = 0.93) in five brands. C, H, N analysis showed C approximately 55%, H approximately 12%, and N approximately 2% with a total of approximately 70% organic matter. However, thermal decomposition studies at 700 degrees C suggest less than 5% nonvolatile metal oxides. Herbal formulations contain minor and trace elements in bioavailable forms that favorably influence glucose tolerance and possibly increase the body's ability to ameliorate development of diabetes. PMID- 17916968 TI - Changed clinical chemistry pattern in blood after removal of dental amalgam and other metal alloys supported by antioxidant therapy. AB - This study aimed to investigate a possible connection between removal of dental amalgam restorations supported by antioxidant therapy and indicative changes of clinical chemistry parameters. A group of 24 patients, referred for complaints related to amalgam restorations, underwent a removal of their amalgams. All patients were treated with antioxidants (vitamin B-complex, vitamin C, vitamin E, and sodium selenite). An age- and sex-matched control group of 22 individuals was also included. The mercury (Hg) and selenium (Se) concentration in plasma, Hg concentration in erythrocytes, and 17 clinical chemistry variables were examined in three groups: patients before amalgam removal (Before), patients after amalgam removal (After), and control individuals (Control). The Hg and Se values decreased (p < 0.05) in plasma, and the Hg concentration decreased (p < 0.05) in erythrocytes after amalgam removal. The variables serum lactate dehydrogenase (serum LDH) and serum sodium differed significantly both when comparing Control with Before (p < 0.01) and Before with After (p < 0.01). The variables white blood cell count (WBC), blood neutrophil count, blood eosinophil count, blood basophil count, blood lymphocyte count, blood monocyte count, serum potassium, and serum creatinine differed in the Before/After test (p < 0.05). Multivariate statistics (discriminant function analysis) could separate the groups Before and After with only one misclassification. PMID- 17916969 TI - Effects of dietary copper (II) sulfate and copper proteinate on performance and blood indexes of copper status in growing pigs. AB - 160 crossbred (Duroc x Landrace x Yorkshire) gilts averaged 21.25 kg body weight were used to study the effects of dietary copper (II) sulfate (CuSO4) and copper proteinate (Cu-Pr) on growth performance, plasma Cu concentration, ceruloplasmin activity, and erythrocyte Cu/Zn-superoxide dismutase (SOD) activity. All pigs were allotted to four treatments and fed with basal diets supplemented with 0 (control), 250 mg /kg Cu as CuSO4, and 50 and 100 mg/kg Cu as Cu-Pr. Growth performance was determined based on two growth phase (phase 1: days 0 to 15, phase 2: days 15 to 30). After 30 days of the treatment, 16 pig blood samples (four per treatment) were collected for indexes of copper status determination. The experimental results showed that compared with control, pigs fed with 250 mg Cu/kg as CuSO4 and 100 mg Cu/kg as Cu-Pr had higher average daily gain and average daily feed intake in the whole growth phase (d 0 to 30). In addition, 250 mg Cu/kg as CuSO4 and 100 mg/kg Cu as Cu-Pr enhanced plasma ceruloplasmin activity (P < 0.05), and 100 mg/kg Cu as Cu-Pr increased erythrocyte Cu/Zn-SOD activity (P < 0.01) compared with the control. There was no obvious treatment response on plasma Cu concentration in the present study. PMID- 17916971 TI - Effect of garlic supplementation on erythrocytes antioxidant parameters, lipid peroxidation, and atherosclerotic plaque formation process in oxidized oil-fed rabbits. AB - Effect of garlic supplementation on blood antioxidant status, lipid peroxidation, and coronary plaque formation process was investigated in oxidized oil-fed rabbits. Eighteen adult male mixed European rabbits were given a balanced diet (21 g% protein, 34 g% fat, 45 g% carbohydrate), which contained isocaloristic addition of nonoxidized or oxidized rapeseed oil in the presence and absence of garlic. The experiment lasted 24 weeks. At the beginning and every 6 weeks, rabbits were weighed, and blood was taken. To evaluate the antioxidant status of the rabbits, erythrocytes malondialdehyde (MDA) concentration, total superoxide dismutase (t-SOD), and glutathione peroxidase (GPX) activations were determined. After the experiment was completed, aortas were dissected for histological examinations. Changes in the contents of the above parameters and histological examinations showed that oxidized rapeseed, oil administered to rabbits, caused the development of atherosclerotic changes and disturbed antioxidant status. The addition of garlic in such diets inhibited atherosclerotic changes in the aorta wall, and it is related to the homeostatic activity of antioxidative enzymes and lipid peroxidation. PMID- 17916970 TI - The influence of methionine, selenomethionine, and vitamin E on liver metabolic pathways and steatosis in high-cholesterol fed rabbits. AB - Significant disorders of liver metabolic pathways enzymes after high-cholesterol diet could give information on liver steatosis development. This process could probably also be inhibited by some compounds, as examined in rabbits. Forty-two male rabbits were served a high-cholesterol diet (2 g%) (0.67 g/kg b.m./24 h) with addition of d,l-methionine (70 mg/kg b.m./24 h) or seleno-d,l-methionine (12.5 microg/kg b.m./24 h) or alpha-tocopherol (10 mg/kg b.m./24 h) for 3 months to compare the protection effect of used compounds on liver metabolism and steatosis. At the beginning and every month, blood was taken. After the experiment was completed, livers were dissected for histological examinations. The concentration of total cholesterol (t-CH), triacylglycerol (TG), and the activities of aldolase (ALD), sorbitol dehydrogenase (SDH), glutamate dehydrogenase (GLDH), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were determined. Plasma t-CH and TG concentrations were significantly higher in all experimental groups vs control group. Blood serum AST and ALT activities did not undergo change but there were observed not significant increase in the CH group vs control group. Activities of SDH, GLDH, and LDH increased in blood serum and decreased in the liver in all experimental groups. Activities of LDH and SDH increased in the liver in the CH+Met group vs CH group. ALD activity decreased in the liver only in the CH and CH+Se groups. This data support a lipotoxic model of cholesterol-mediated hepatic steatosis. Prolonged administration of high-cholesterol diet not only disturbs the structure of cell membranes, which is expressed by decreased activity of enzymes in the liver and the migration of those enzymes to plasma but as well leads to steatosis of the liver, which has been confirmed by histological examinations. The applied compounds appear to have a varying influence upon the activity of enzymes determined in serum and liver. Obtained results showed a beneficial influence of methionine and vitamin E supplementation on liver steatosis development. PMID- 17916972 TI - Protective effects of antioxidants against cadmium-induced oxidative damage in rat testes. AB - The protective effects of melatonin, vitamin E, and selenium alone or in combination were tested against cadmium-induced oxidative damage in rat testes. A total of 60 male rats were equally divided into five study groups, one of which acted as control receiving subcutaneous injections of physiological saline. The remaining four groups were treated with subcutaneous injections of cadmium chloride at a dose of 1 mg/kg weight. The first study group received no treatment. The second group was treated with a combination of 60 mg/kg vitamin E and 1 mg/kg sodium selenite. Group 3 was treated with 10 mg/kg melatonin, and the fourth group received a combination of vitamin E, sodium selenite, and melatonin at the doses mentioned above. After 1 month, the animals were killed, and the testes were excised for histological inspection and determination of tissue malondialdehyde and the activity of superoxide dismutase. The animals receiving no treatment showed significantly higher malondialdehyde levels and reduced activity of the enzyme (p < 0.05). Treatment with antioxidants resulted in a significant reduction in malondialdehyde when compared to the nontreated animals (p < 0.05) and an increase in the superoxide dismutase activity that was almost the same as the controls. The combination of melatonin, vitamin E, and selenium appears to have the more profound effect against cadmium-induced testicular injury. PMID- 17916973 TI - Protective effect of selenium on nicotine-induced testicular toxicity in rats. AB - Effect of exogenous selenium at a dose of 10 mug/kg body weight on the testicular toxicity induced by nicotine in rats was investigated. Male albino rats were maintained for 60 days as follows: (1) control group (normal diet), (2) nicotine group (0.6 mg /kg body weight), (3) selenium (10 microg/kg body weight), and (4) nicotine (0.6 mg/kg body weight) + selenium (10 microg/ kg body weight). Administration of nicotine caused reduction in sperm count and sperm motility. Activity of HMG CoA reductase and concentration of cholesterol were increased in the testes of the nicotine administered group. Activities of testicular enzymes 3beta hydroxysteroid dehyrogenase (3 betaHSD), 17beta hydroxysteroid dehyrogenase (17 betaHSD) were decreased. Levels of testosterone in the serum were also reduced. However, the extent of these alterations was lesser in the group administered with nicotine along with selenium. Analysis of plasma revealed reduced quantity of cotinine in the group co-administered with nicotine along with selenium in comparison with the nicotine group. Nondetectable levels of nicotine were present in the co-administered group. This indicates altered metabolism of nicotine when administered along with selenium. PMID- 17916974 TI - 131I induced hematological alterations in rat blood: protection by zinc. AB - The present study was planned to determine the potential of zinc in attenuating the toxicity induced by 131I in rat blood. Female wistar rats were segregated into four main groups. Animals in Group I served as normal controls; Group II animals were administered a dose of 3.7 Mbq of 131I (carrier free) intraperitoneally, Group III was supplemented with Zinc in the form of ZnSo4.7H2O (227 mg/l drinking water), and Group IV was given a combined treatment of Zinc as well as 131I, in a similar way as was given to Groups IV and II animals, respectively. The effects of different treatments were studied on various parameters in rat blood including hemoglobin (Hb) levels, % hematocrit, zinc protoporphyrins (ZPP), activities of enzymes which included aminolevulinic acid dehydratase (delta-ALAD) and Na+ K+ ATPase and uptake of 65Zn in blood. The study revealed an increase in the levels of hemoglobin, % hematocrit, activities of delta-ALAD, Na+ K+ ATPase and uptake of 65Zn, 7 days after the 131I treatment. On the contrary, the levels of ZPP were found to be significantly decreased after 131I treatment. However, zinc treatment to 131I-treated animals significantly attenuated the various biochemical and hematological indices. Moreover, zinc treatment to the 131I-treated animals could significantly decrease the uptake of 65Zn, which was increased after 131I treatment. Based upon these data, the present study suggests that zinc has the potential to attenuate 131I induced toxicity by restoring the altered hematological indices and biochemical changes. PMID- 17916975 TI - Garlic (Allium sativum L.) as a potential antidote for cadmium and lead intoxication: cadmium and lead distribution and analysis in different mice organs. AB - Analysis and distribution of Pb and Cd in different mice organs including liver, kidney, spleen, heart and blood were evaluated after treatment with different aqueous concentrations of garlic (12.5-100 mg/l). Atomic absorption spectrometry (AAS) was used for analysis of Pb and Cd in these organs. Treatment of Cd-Pb exposed mice with garlic (12.5-100 mg/l) reduced Pb concentrations by 44.65, 42.61, 38.4, 47.56, and 66.62% in liver, kidney, heart, spleen and blood respectively. Moreover, garlic reduced Cd levels by 72.5, 87.7, 92.6, 95.6, and 71.7% in liver, kidney, heart, spleen and blood respectively. The suppressed immune responses in mice pretreated with Cd-Pb mixture were reversed by 48.85, 55.82, 81.4 and 90.7 in the presence of 100, 50, 25, and 12.5 mg/ml of garlic extract. PMID- 17916976 TI - Essential metal status, prooxidant/antioxidant effects of MiADMSA in male rats: age-related effects. AB - Thiols are known to act as protectants in the biological system for their involvement in a number of metabolic regulations. In this study, we investigated the effect of a new and potent thiol-chelating agent, monoisoamyl 2,3 dimercaptosuccinic acid (MiADMSA), an analog of meso 2,3-dimercaptosuccinic acid, to find out if it could act as a prooxidant (because of its lipophilic character) or antioxidant (because of thiol moiety) that could supplement its chelating properties in different age groups of male rats (young, adult, and old rats) and produce effective clinical recoveries in the treatment of metal intoxication. Animals were treated with 25, 50, and 100 mg/kg of MiADMSA, i.p, once daily for 1 week to assess the effect on the antioxidant system in major organs based on sensitive biochemical variables indicative of oxidative stress. Results suggested that MiADMSA administration increased the activity of d-aminolevulinic acid dehydratase in all the age groups and increased blood glutathione (GSH) levels in young rats. MiADMSA also potentiated the synthesis of metallothioneine in liver and kidneys and GSH levels in liver and brain. Apart from this it also significantly reduced the glutathione disulfide levels in tissues. However, administration of MiADMSA caused some concern over the copper loss. MiADMSA was found to be safe in rats of all ages. PMID- 17916977 TI - Effects of nanoparticles on the adhesion and cell viability on astrocytes. AB - In recent years, both pharmaceutical companies and manufacturing industries have expressed heightened interest in the potential applications of magnetic nanoparticles for therapeutic and technological purposes. Specifically, pharmaceutical companies seek to employ magnetic nanoparticles as carriers to facilitate effective drug delivery, especially in areas of the brain. Manufacturing industries desire to use these nanoparticles as ferrofluids and in magnetic resonance imaging. However, data concerning the effects of magnetic nanoparticles on the nervous system is limited. This study tested the hypotheses that nanoparticles can (1) inhibit adherence of astrocytes to culture plates and (2) cause cytotoxicity or termination of growth, both end points representing surrogate markers of neurotoxicity. Using light microscopy, changes in plating patterns were determined by visual assessment. Cell counting 4 days after plating revealed a significant decrease in the number of viable astrocytes in nanoparticle treated groups (p < 0.0001). To determine the cytotoxic effects of nanoparticles, astrocytes were allowed to adhere to culture plates and grow to maturity for 3 weeks before treatment. Membrane integrity and mitochondrial function were measured using colorimetric analysis lactate dehydrogenase (LDH) and 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTS), respectively. Treatment with nanoparticles did not significantly alter astrocytic LDH release (p > 0.05) in the control group (100% +/- 1.56) vs the group receiving treatment (97.18% +/- 2.03). However, a significant increase in MTS activity (p < 0.05) between the control (100% +/- 3.65) and treated groups (112.8% +/- 3.23) was observed, suggesting astrocytic mitochondrial uncoupling by nanoparticles. These data suggest that nanoparticles impede the attachment of astrocytes to the substratum. However, once astrocytes attach to the substratum and grow to confluence, nanoparticles may cause mitochondrial stress. PMID- 17916978 TI - Reduction of aluminum toxicity by 2-isopropylmalic acid in the budding yeast Saccharomyces cerevisiae. AB - The budding yeast Saccharomyces cerevisiae secretes 2-isopropylmalic acid (2 iPMA), an intermediate in leucine biosynthesis. Because 2-iPMA binds Al(III) in the culture medium, it is thought to reduce toxicity by Al(III). The effects of 2 iPMA and malic acid (MA) on Al toxicity were investigated in a medium with a low pH and low concentrations of phosphates and magnesium. The reduction in the growth of S. cerevisiae observed in the presence of 100 muM Al(III) ions was relieved more by the addition of 1.0 mM 2-iPMA than by 1.0 mM MA, indicating that 2-iPMA possesses superior Al(III)-ion detoxification ability. Investigations using the wild type and the deltaleu4 and deltaleu9 mutant strains indicated that secretion of a sufficient level of 2-iPMA was required to enhance the Al tolerance. It is thought that 2-iPMA secreted from the yeast cells chelates Al ions and prevents them from entering the cells, resulting in Al tolerance. PMID- 17916979 TI - Exposure of Dunaliella tertiolecta to lead and aluminum: toxicity and effects on ultrastructure. AB - The growth response of the marine alga Dunaliella tertiolecta to different concentrations of lead and aluminum was investigated. Both metals had a stimulatory effect at low concentration and an inhibitory effect at high concentration (hormesis). The IC25 values of lead are 8.43, 7.29, and 6.74 mg L-1 for 24, 48, and 72 h, respectively. The corresponding values for aluminum are 30.54, 22.42, and 18.16 mg L-1. Although it seems that the two metals are not directly toxic to the alga at the concentrations found in the environment, as implied by the IC25 values and the environmental concentrations of the metals, low concentrations of both metals, alone and in combination, affected the ultrastructure. The growth of batch-grown cells exposed to 0.5 mg L-1 lead and aluminum, alone and combined, during the 24-h exponential phase was investigated. The same cells were also examined under an electron microscope to determine the biological effects of the two metals on the ultrastructure. The most obvious effects of lead were disrupted thylakoidal membranes, accumulated polyphosphate bodies and vacuoles, and lead precipitates on the cell surface. These ultrastructural alterations were partially present in aluminum-treated and lead aluminum-treated cells. In joint exposure, the most important change was the lysis of the cell membrane. Aluminum and lead seem to act synergistically on the cell membrane leading to cell membrane lysis. PMID- 17916980 TI - Effects of nano-anatase on spectral characteristics and distribution of LHCII on the thylakoid membranes of spinach. AB - In the article, we report that effects of nano-anatase on the spectral characteristics and content of light-harvesting complex II (LHCII) on the thylakoid membranes of spinach were investigated. The results showed that nano anatase treatment could increase LHCII content on the thylakoid membranes of spinach and the trimer of LHCII; nano-anatase could enter the spinach chloroplasts and bind to PSII. Meanwhile, spectroscopy assays indicated that the absorption intensity of LHCII from nano-anatase-treated spinach was obviously increased in the red and the blue region, fluorescence quantum yield near 685 nm of LHCII was enhanced, the fluorescence excitation intensity near 440 and 480 nm of LHCII significantly rose and F 480/F 440 ratio was reduced. Oxygen evolution rate of PSII was greatly improved. Together, nano-anatase promoted energy transferring from chlorophyll (chl) b and carotenoid to chl a, and nano-anatase TiO2 was photosensitized by chl of LHCII, which led to enhance the efficiency of absorbing, transferring, and converting light energy. PMID- 17916981 TI - APS--more systemic disease than SLE. AB - The antiphospholipid syndrome is a systemic autoimmune disease that can have serious consequences for patients. Importantly, there is a wide range of clinical presentations. In this issue we have attempted to provide an overview of these features and place it in the context of autoimmunity. PMID- 17916982 TI - Clinical thrombotic manifestations in SLE patients with and without antiphospholipid antibodies: a 5-year follow-up. AB - OBJECTIVE: To analyze the association of antiphospholipid antibodies (aPL) with the development of clinical thrombotic manifestations and to characterize the efficacy of anti-thrombotic therapies used. METHODS: 272 systemic lupus erythematosus (SLE) patients participated in the study. Patient files and a cumulative database were used to collect patients' medical histories. Anti cardiolipin (aCL), anti-beta2-glycoprotein I (abeta2GPI) antibodies, and lupus anticoagulant (LAC) were measured according to international recommendations. New thrombotic events were registered during follow-up. RESULTS: The patients were prospectively studied for 5 years, of whom 107 were aPL negative (aPL- group). Criteria for antiphospholipid syndrome (APS) were fulfilled by 84 of 165 aPL positive patients (APS+ group) indicating that SLE patients with aPL have around 50% risk to develop thrombotic complications. The aPL+ group (n = 81) consisted of aPL+ but APS- patients. LAC was the most common aPL (n = 27, 32.1%) in patients with APS. The cumulative presence of aPL further increased the prevalence of thrombotic events. During the follow-up period, aPL developed in 8 of 107 patients (7.5%) from the aPL- group, of whom 3 (2.8%) presented with thrombotic complications. Other types of aPL developed in 7 of 165 (4.2%) aPL+ patients within 5 years. New thrombotic events occurred in 3.7% of aPL+ (n = 3) and 8.3% (n = 7) of the APS group. During follow-up, 52 of 81 aPL+ patients received primary prophylaxis, and 1 (1.9%) had transient ischemic attack (TIA). In the non-treatment group, 2 (6.9%) had stroke. Seventy-nine of 84 of the APS patients received secondary prophylaxis, and myocardial infarction occurred in 2 patients (on cumarine therapy maintaining an international normalized ratio around 2.5-3.0), and 5 suffered a stroke/TIA (1 on aspirin and 4 on aspirin + cumarine). CONCLUSION: The findings emphasize the importance of determining both aCL and abeta2GPI antibodies and LAC in SLE patients and the need for adequate anticoagulant therapy. PMID- 17916983 TI - Livedo reticularis as a criterion for antiphospholipid syndrome. AB - Many consensus meetings were organized in an attempt to improve the present criteria for antiphospholipid syndrome (APS) classification. In this regard, a high prevalence of antiphospholipid antibodies in systemic lupus erythematosus patients was reported in association with the presence of livedo reticularis (LR). In these studies, the association between LR, migraine, and the development of thrombosis (strokes, valvular dysfunctions) was evident. During the last decade, it was strongly suggested that many clinical symptoms (LR, valvular dysfunctions) or laboratory features (thrombocytopenia) should be considered as "minor criteria" for APS. The inclusion of these clinical symptoms in the criteria for APS classification could become of additive value especially when they exist together in one patient. This review summarizes the data that question or support this idea. PMID- 17916985 TI - Pulmonary hypertension, antiphospholipid antibodies, and syndromes. AB - Antiphospholipid antibodies have been associated with two types of pulmonary hypertension (PHT), the thromboembolic type, after deep venous thromboses in the lower limbs complicated by pulmonary embolism and the "primary" plexogenic type. The PHT may occur in the absence of any other manifestations of the antiphospholipid syndrome (APS), and cases have been recorded with very high levels of antiphospholipid antibodies. It may also accompany systemic lupus erythematosus (SLE) and may manifest with or without other features of the APS. It may also form part of the clinical presentation of a "primary" APS. Its prevalence is of the order of 1.8-3.5% of the manifestations of the APS depending on the series. Primary "idiopathic" PHT has long been regarded as an "immunological" disorder. Its manifestations are essentially to the primary type seen with the connective tissue disorders such as SLE, APS, mixed connective tissue disease, calcinosis, Raynaud phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia variety of systemic sclerosis and Sjogren's syndrome. The high prevalence of PHT in patients with human immunodeficiency virus infection who demonstrate low CD4 counts points to a close relationship between the T regulatory cells (Treg) and the development of PHT, and this hypothesis is discussed in this review. Genetic and chromosomal aspects of PHT are also discussed. PMID- 17916986 TI - Anti-phospholipid antibodies and infertility. AB - Antiphospholipid syndrome (APS) or the presence of antiphospholipid antibodies (aPL), usually presents as pregnancy loss. However, aPL have also been reported to affect implantation, placentation, and early embryonic development. The binding of aPl to beta2GP1 may lead to breakdown of the phospholipid adhesion molecules between different elements of trophoblast. As aPL affect placental growth and function, aPl may prevent implantation presenting as infertility. Lupus anticoagulant and anticardiolipin antibody have been implicated in the prothrombotic effects of APS. Antibodies to other phospholipids such as anti phosphatidylserine, phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl glycerol, phosphatidyl Inositol etc. may be more relevant in infertility. Their role remains to be clarified. There is theoretical evidence from animal models and clinical infertility practice that aPL has a role in infertility. However, a large-scale meta-analysis has failed to confirm the association. To determine whether infertility or even pregnancy loss is associated with aPL, it is necessary to know that the embryo is chromosomally normal. Pregestational diagnosis has shown that up to 60% of embryos may be chromosomally aneuploid in failed in vitro fertilization (IVF); hence, may confound our understanding concerning the association between aPL and infertility, failed IVF or even pregnancy loss. PMID- 17916987 TI - Primary anetoderma and antiphospholipid antibodies--review of the literature. AB - Primary anetoderma (PA) is a rare elastolytic disorder that develops in clinically normal skin or following a nonspecific inflammatory process. The old literature contains numerous reports of the association between PA and lupus eryhtematosus, although the relationship has not been clearly established. In recent years, there has been a growing body of literature linking PA with a wide range of immunologic abnormalities, the most common of which is the presence of antiphospholipid antibodies, with or without antiphospholipid syndrome. The present review summarizes the literature, from the early descriptions pointing toward an immunologic basis of PA and up to the present recognition that PA is a cutaneous sign for autoimmune disorders, in general, and the presence of antiphospholipid in particular. PMID- 17916984 TI - Novel insights into associations of antibodies against cardiolipin and beta2 glycoprotein I with clinical features of antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is a clinical autoimmune disorder characterized by arterial and/or venous thrombosis and/or pregnancy morbidity, associated with the persistence of lupus anticoagulant or anticardiolipin (aCL) antibodies. Accumulating evidence indicates that phospholipid binding protein, beta2 glycoprotein I (beta2GPI) represents the major target antigen for antiphospholipid (aPL) antibodies and plays a role in the pathogenesis of APS. It is widely accepted that aPL antibodies detected by conventional solid phase assays in patients with APS are mainly directed against a complex of aCL and anti beta2GPI, although antibodies against beta2GPI protein can now also be detected by specific ELISA using purified proteins in solid phase. Despite the fact that these antibodies are not listed in the new diagnostic criteria, a high specificity of anti-beta2GPI assay for the clinical features of APS was established. During the last decade, numerous studies have investigated the clinical link between aCL and/or anti-beta2GPI antibodies and diverse features of APS. This manuscript reviews the current studies published recently in this field and discusses the relationship between the existence of aCL and anti-beta2GPI antibodies and the main and unusual manifestations of APS. PMID- 17916988 TI - Orthopedic involvement in antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is a common autoimmune disease, manifested by vascular thrombosis and fetal loss in the presence of antiphospholipid antibodies. Orthopedic involvement is a relatively novel and under-recognized feature of APS. In this article we review the association of primary, secondary, and catastrophic APS with diverse orthopedic conditions, including osteonecrosis in adult and pediatric patients, bone marrow necrosis, nontraumatic fractures, and some other disorders. PMID- 17916989 TI - Systemic antiphospholipid syndrome and atherosclerosis. AB - Atherosclerosis (AT) is a metabolic, systemic inflammatory/immune disease characterized by lipoproteins metabolism alteration that leads to immune/inflammatory system activation with the consequent proliferation of smooth muscle cells, narrowing arteries and atheroma formation. Antiphospholipid syndrome (APS) is a systemic autoimmune disease characterized by thrombophilic state and circulating antiphospholipid antibodies (aPL) including anti beta2-GPI. Experimental studies and human observations suggest that APS is associated with AT. In fact, innate and adaptive immune responses participate in the pathogenesis of both diseases. Anti-oxLDL, anti-aPL, anti beta2GPI, anti-HSP antibodies, among others, has been found in patients with APS and AT. Endothelial dysfunctions, oxidative stress, increase of cell adhesion molecules, active platelets, are common findings in both diseases. Macrophages, dendritic cells, T-cell activation, CD40-CD40 ligand interaction, are considered as pathogenic mechanism of AT and APS. Premature AT may be the first symptom of APS. Thrombophilia, aPL antibodies, and APS may be present in patients with premature AT. An association between AT and venous thrombosis (a clinical hallmark of APS) has been proposed in unselected patients with deep venous thrombosis of the legs without symptomatic AT. Asymptomatic AT, defined in terms of carotid intima media thickness and lumen diameter decrease, was observed in patients with APS. Premenopausal female patients with PAPS have a higher prevalence of cerebrovascular disease in comparison with male patients. Accelerated AT and hormones could be the explanation of these findings. High levels of aCLs, significantly predict the risk of future ischemic stroke in women but not in men. AT is one of the main features of systemic APS and offer opportunities for new treatment strategies. PMID- 17916990 TI - The Heart and APS. AB - The heart is a target organ in antiphospholipid syndrome (APS). Endocardial disease, intracardiac thrombosis, myocardial involvement including coronary heart disease and microvascular thrombosis, as well as pulmonary hypertension have all been described in APS patients. Valvular involvement is the most common manifestation with a prevalence of 82% detected by transesophageal echocardiography. Symmetrical, nodular thickening of the mitral and/or aortic valves is characteristic. Anticoagulant/antiplatelet treatment is ineffective in terms of valvular lesion regression. Some patients require cardiac valve replacement. However, patients with APS have shown an increased perioperative morbidity and mortality. Intracardiac thrombosis, although a rare complication, can cause pulmonary and systemic emboli. Differential diagnosis with myxoma may be very difficult. PMID- 17916991 TI - Antiphospholipid syndrome and cancer. AB - Thrombosis is a frequent complication of cancer that is a substantial cause of morbidity and mortality. The association of antiphospholipid antibodies (aPL) and cancer has been under investigation for several years. Recent findings suggest an increased prevalence of certain cancers in aPL-positive patients; thus, an intensive search for an occult malignancy is prompted in these patients. In addition, several studies reported on elevated levels of aPL in various malignancies; it seems, however, that aPL levels do not reflect their pathogenicity; therefore, their pathological significance in these subset of patients is still elusive. Continuing research on the association between the antiphospholipid syndrome/aPL and malignancies is important, given the potential impact on the understanding and treatment of both antiphospholipid syndrome and cancer. PMID- 17916992 TI - Antiphospholipid syndrome and cognition. AB - In addition to the well-defined neurologic events due to arterial and venous thrombotic vascular occlusions of antiphospholipid syndrome (APS), a broad spectrum of neuropsychiatric has been related to antiphospholipid (aPL). Experimental evidence of a pathogenic role of aPL in mice with impaired neurological function disclosed inflammatory reaction as a hallmark. The process that leads to neurological dysfunction seems to be both structurally destructive and functionally impairing. The most modern resources of neuroimmaging also suggest that, in addition to the micro-infarcts that occur in strategic areas, other metabolic impairments are related to progressive dementia and aPL presence. Although there is a lot of confusion among APS and lupus' cognitive involvement, there is a body of experimental and clinical evidence that aPL causes this kind of damage. PMID- 17916993 TI - Looking into the eyes of patients with antiphospholipid syndrome. AB - Diagnosis of antiphospholipid syndrome (APS) should be considered in all patients with recurrent systemic or ocular thrombosis in the absence of known risk factors. Because of the frequent ocular involvement in APS patients (as many as 80%), an ophthalmologic assessment should become a routine part of the clinical work-up of all patients in whom APS is highly suspected. The presence of isolated ocular thrombosis with persistently increased titers of antiphospholipid antibodies should be considered as definite APS. Ocular involvement in APS is frequently associated with other manifestations of the central nervous system (CNS), such as transient ischemic attack or cerebral vascular events. Diagnostic tools are needed to better establish a diagnosis of retinal vascular thrombosis. The treatment of isolated ocular APS should be based on the same principles as in all patients with systemic APS. Anticoagulation is aimed to prevent recurrent ocular or cerebral thromboses. PMID- 17916995 TI - The cytoplasmic expression of MUC1 in papillary thyroid carcinoma of different histological variants and its correlation with cyclin D1 overexpression. AB - This study addressed the immunohistochemical expression of MUC1 in papillary thyroid carcinoma (PTC) of different histotypes, sizes, and morphological features of aggressiveness, and its correlation with the overexpression of cyclin D1, a target molecule of the Wnt pathway. MUC1 expression was examined in a total of 209 PTCs. Cytoplasmic MUC1 expression was elevated in the tall, columnar cell and oncocytic variants (100%), Warthin-like (78%), and conventional PTCs (61%), and in papillary microcarcinoma (PMC) with the conventional growth pattern (52%). On the contrary, it was low in the follicular variant (27%) of PTC and PMCs with follicular architecture (13%). Cytoplasmic MUC1 accumulation did not associate with any clinicopathological features except peritumoral lymphoid infiltration in PTCs and in PMCs with the conventional growth pattern. MUC1 staining correlated with cyclin D1 overexpression in conventional PTCs and PMCs and PMCs with follicular architecture. The results demonstrate that MUC1 expression varies broadly in different histological variants of PTC, being the lowest in tumors with follicular structure. In general, it does not prove to be a prognosticator of PTC aggressiveness. A high correlation between MUC1 and cyclin D1 implies MUC1 involvement in the Wnt cascade functioning in a large subset of human PTCs and PMCs. PMID- 17916996 TI - AIP Mutations are not identified in patients with sporadic pituitary adenomas. AB - The pathogenesis of pituitary adenomas remains a subject of interest. Recently, mutations in the aryl hydrocarbon receptor-interacting protein (AIP) were identified as germline events leading to pituitary tumor predisposition in Finnish and Italian families with familial growth hormone-secreting pituitary adenomas and acromegaly. We examined the frequency of AIP mutations in pituitary tumors and blood of Canadian patients with sporadic pituitary somatotroph adenomas and sporadic pituitary adenomas of other types. Genomic DNA was extracted from pituitary tumors and white blood cells obtained from peripheral blood. Three PCR reactions were carried out to amplify the sites of known mutation, and amplified products were sequenced. AIP mutations were not detected as germline events in blood or as somatic alterations in tumors of 66 patients with pituitary adenomas. These included 50 acromegalics and 16 patients with other types of pituitary tumor. No mutations were detected in the blood of 22 controls and 10 patients with other endocrinopathies. Our results indicate that mutations in AIP are not identified in sporadic pituitary adenomas of Canadian patients. This rare mechanism of pituitary tumorigenesis appears to be unique to the initial Finnish and Italian families described. PMID- 17916994 TI - RET signaling in endocrine tumors: delving deeper into molecular mechanisms. AB - The rearranged during transfection (RET) proto-oncogene encodes a receptor tyrosine kinase that is implicated in the development of endocrine tumors of the thyroid and adrenal glands. In humans, activating RET mutations are found in the inherited cancer syndrome multiple endocrine neoplasia 2 and in sporadic medullary and papillary thyroid carcinomas. The specific type and location of RET mutations are strongly correlated with the disease phenotype and have both diagnostic and prognostic value. Recent advances in the molecular characterization of the RET receptor and its mutants have begun to define the mechanisms underlying the transforming ability of the different RET mutant forms. This information has revealed key functional features of these mutant proteins that distinguish the different clinically recognized mutations and provide clues as to the functional origins of the phenotypes associated with specific RET mutations. The elucidation of molecular mechanisms involved in RET-mediated transformation is a key step in the development of much needed therapeutics that target RET's oncogenic properties. Recent advances have begun to provide a deeper understanding of the receptor's function, and dysfunction, in human tumors that may guide this process. PMID- 17916997 TI - Differential expression of the five somatostatin receptor subtypes in human benign and malignant insulinomas - predominance of receptor subtype 4. AB - Insulinomas constitute a subgroup of pancreatic endocrine tumors showing B cell differentiation and clinical symptoms related to inappropriate insulin secretion (WHO). Many endocrine tumors express somatostatin receptors (sstrs), which can be visualized by octreotide scintigraphy; however, about half of all insulinomas are reported to be negative. Previous immunohistochemical investigations with antibodies to sstr subtypes 1, 2, 3, and 5 have revealed differences in expression between various neuroendocrine tumors. In the present study, the immunoreactivity to all five human sstr was studied in ten benign and six malignant human insulinomas. Sstr4 was the receptor subtype most frequently expressed in both benign and malignant tumors. A difference in the immunohistochemical sstr5 expression pattern was seen between benign and malignant tumors: Three of the six malignant tumors, but none of the benign tumors, expressed sstr5. The other receptor subtypes were expressed in low numbers with no difference between benign and malignant tumors. The finding of a strong expression of sstr4 in both benign and malignant insulinomas suggests that this receptor subtype could be of importance for diagnostic and therapeutic use. PMID- 17916998 TI - Pituitary adenoma with tumoral granulomatous reaction. AB - Herein, we report a unique case of an adult male with a corticotrophic pituitary adenoma of silent subtype 1 exhibiting conspicuous idiopathic tumoral noncaseating granulomatous inflammation. The lesion was unassociated with clinical or laboratory evidence of either systemic sarcoidosis or infection. Histochemical and polymerase chain reaction (PCR) studies revealed neither fungi nor tubercle bacilli. We suggest that tumoral production of an as yet uncharacterized antigen may have induced the granulomatous inflammatory reaction. PMID- 17916999 TI - A large sellar granular cell tumor in a 21-year-old woman. AB - We report here the case of a 21-year-old woman with a large sellar tumor, extending to the suprasellar area associated with growth hormone deficiency, hypogonadism, hypocorticism, and hyperprolactinemia. Transsphenoidal surgery was performed, and histologic, immunohistochemical, and electron microscopic study lead to the diagnosis of granular cell tumor. These tumors are, in most cases, very small and are found incidentally at autopsy of older patients. Our case is exceptional because the tumor developed in a young woman, extended to the suprasellar region, and caused clinical symptoms. PMID- 17917000 TI - Cushing's syndrome in a child with pancreatic acinar cell carcinoma. AB - A case of pancreatic acinar cell tumor (ACC) is presented in a 10-year-old boy. The tumor manifested clinically with Cushing's syndrome, high serum adrenocorticotropic hormone (ACTH) and cortisol concentrations. In addition, excessive serum levels of alpha-fetoprotein (AFP) were detected. Surgical resection was not possible due to retroperitoneal invasion. Biopsy of the mass showed a solid, poorly differentiated ACC of the pancreas. Periodic acid Schiff positive cytoplasmic granules, trypsinogen, keratins, alpha-1-antitrypsin, and AFP were identified in the tumor cells. Electron microscopy demonstrated zymogen granules as well as isolated dense core granules. Using immunochemiluminometric assay, a high quantity of ACTH was found in the fresh frozen tumor extract. ACTH, chromogranin A, and corticotropin-releasing factor were identified only in a few cells by immunohistochemistry. Combined radiochemotherapy was temporarily effective in reducing the tumor mass and serum AFP. Serum ACTH and cortisol levels dropped progressively and definitively to normal values after chemotherapy, and the Cushing's syndrome subsided. Two years later, the patient died with metastatic disease. The presented case of ACC is interesting due to high serum AFP values and ectopic ACTH secretion resulting in Cushing's syndrome. PMID- 17917003 TI - Introduction: Robert A. Good Immunology Conference 2007. PMID- 17917001 TI - Myelolipoma with adrenocortical adenoma: an unusual combination that can resemble carcinoma. AB - Myelolipoma is a benign tumor that occurs in the adrenal gland and rarely in extra-adrenal sites. Commonly, it is diagnosed as an incidental finding. However, some reports indicate synchronous occurrence with other lesions including adrenocortical carcinoma. It has very rarely been reported to occur in association with adrenocortical adenoma. In this paper, we report three cases of adrenal myelolipoma associated with adrenocortical adenoma; in all three patients, the radiological appearance resembled adrenocortical carcinoma. These cases emphasize the importance of this combination as a pitfall in the correct diagnosis and management of patients with adrenal masses. PMID- 17917007 TI - The impact of physician education and public awareness on early diagnosis of primary immunodeficiencies: Robert A. Good Immunology Symposium. PMID- 17917008 TI - Immunology today and new discoveries: building upon legacies of Dr. Robert A. Good. PMID- 17917009 TI - Robert A. Good, the March of Dimes, and immunodeficiency: an historical perspective. AB - Dr. Robert A. Good and the March of Dimes Birth Defects Foundation maintained a close association for a quarter century in the fight against immunodeficiency diseases. The March of Dimes, whose mission is to prevent birth defects, premature birth, and infant mortality, awarded an initial grant to Dr. Good in 1960 to conduct basic clinical and experimental studies on arthritis and collagen diseases. By 1966, this support broadened to include Dr. Good's research on agammaglobulinemia, ataxia telangiectasia, Chediak-Higashi disease, and Wiskott Aldrich syndrome. Dr. Good led three historic March of Dimes conferences on immunodeficiency and, in 1968, conducted the first bone marrow transplant to correct an immunological birth defect, memorialized by the March of Dimes in its educational film, Decision (1970). March of Dimes grants to Dr. Good for his research in cellular engineering to genetically correct the defined birth defects approached $1 million for the period 1960-1985. PMID- 17917005 TI - Acquired immunologic tolerance: with particular reference to transplantation. AB - The first unequivocally successful bone marrow cell transplantation in humans was recorded in 1968 by the University of Minnesota team of Robert A. Good (Gatti et al. Lancet 2: 1366-1369, 1968). This achievement was a direct extension of mouse models of acquired immunologic tolerance that were established 15 years earlier. In contrast, organ (i.e. kidney) transplantation was accomplished precociously in humans (in 1959) before demonstrating its feasibility in any experimental model and in the absence of a defensible immunologic rationale. Due to the striking differences between the outcomes with the two kinds of procedure, the mechanisms of organ engraftment were long thought to differ from the leukocyte chimerism associated ones of bone marrow transplantation. This and other concepts of alloengraftment and acquired tolerance have changed over time. Current concepts and their clinical implications can be understood and discussed best from the perspective provided by the life and times of Bob Good. PMID- 17917010 TI - "Experiments of Nature" and the New Era of Immunology: an historical perspective. AB - This review of the events leading to the New Era of Immunology focuses on the important leads provided by the Experiments of Nature. Not only did they provide the path that led to our present knowledge of the ontogeny of the immune system, but also to an entirely new perspective of lymphocytic malignancies. The power of taking clinical clues to the laboratory to conduct experiments not otherwise envisioned and then returning with that information to impact the clinical problem that motivated the studies is elaborated. Finally, the ongoing utility of this approach is emphasized. PMID- 17917011 TI - Severe combined immunodeficiency: new advances in diagnosis and treatment. AB - Severe combined immunodeficiency (SCID) is a condition characterized by lack of cellular and humoral immunity. Uniformly fatal before 1968, SCID was first cured by allogeneic bone marrow transplantation (BMT). Despite improvements in BMT, particularly for cases in which there is no matched related donor, difficulties in SCID treatment persist. Because of incomplete reconstitution and transplant related complications, gene therapy has been pioneered in SCID with success, but also adverse events in the form of leukemic proliferations related to retroviral insertional mutagenesis. Infectious complications are a major limitation to effective treatment. Early diagnosis of SCID in the pre-symptomatic period could be achieved by population-based newborn screening. PMID- 17917012 TI - Immunodeficiencies due to defects of class-switch recombination. AB - Maturation of the antibody response is dependent on class-switch recombination (CSR) and somatic hypermutation (SHM), that modify the structure and the affinity of immunoglobulins, respectively. The cellular and molecular mechanisms involved in these processes have long remained obscure. During the last years, careful investigation of a cohort of rare patients with defective antibody responses have led to the identification of several genes that are critically involved in CSR and SHM. At the same time, recognition that defective maturation of antibody responses may result from different mechanisms, has been essential to better define prognosis and to tailor more appropriate and specific forms of treatment. PMID- 17917013 TI - Common variable immune deficiency: reviews, continued puzzles, and a new registry. AB - BACKGROUND: Common variable immune deficiency (CVID) is a clinically and immunologically heterogenous primary immune deficiency first described more than 50 years ago. The main features are hypogammaglobulinemia, recurrent infections, and other complications. While CVID is considered as a genetic immune defect, and several genes have been reported as leading to the CVID phenotype, one of the most puzzling features of CVID is the sporadic inheritance pattern and the relatively late onset. In most cases, no other family members have any immune defect. The mean age at diagnosis is between 25 and 45 years of age. These features suggest the interplay between either several or numerous genes with or without potential environmental factors. PMID- 17917014 TI - Immunodeficiency, radiosensitivity, and the XCIND syndrome. AB - Through the analysis of a rare disorder called ataxia-telangiectasia (A-T), many important biological lessons have been gleaned. Today, it is clear that the underlying defect of A-T lies in the nucleus, as an inability to repair or process double strand breaks. More important, by the A-T phenotype now allows us to appreciate a much more general distinction between immunodeficiencies that are radiosensitive and those that are not. PMID- 17917015 TI - TACI, isotype switching, CVID and IgAD. AB - Common Variable Immunodeficiency (CVID) is the most prevalent human primary immunodeficiency requiring medical attention. Until recently the only known genetic defect specific to CVID was ICOS deficiency that accounts for about 1% of the patients analyzed. Mutations in the TNFR family member TACI (transmembrane activator and calcium-modulator and cyclophilin ligand interactor), which mediates isotype switching in B cells, were found to be present in 5% of patients with CVID. Mutations in TACI were also found in relatives of patients with CVID who suffered from IgA deficiency (IgAD) as well as in a patient with isolated IgAD. In the majority of patients described to date only one TACI allele is mutated, showing an autosomal dominant transmission of the disease. B cells from individuals with TACI mutations did not produce IgG and IgA in response to the TACI ligand, APRIL (a proliferation-inducing ligand), probably reflecting impaired isotype switching. These results suggest that TACI mutations can lead to CVID. PMID- 17917016 TI - IPEX, FOXP3 and regulatory T-cells: a model for autoimmunity. AB - FOXP3 is the key mediator of regulatory T-cell development in the thymus. Naturally occurring mutations of FOXP3 interfere with this process, resulting in the generation of autoaggressive lymphocyte clones that are directly responsible for the syndrome Immune Dysregulation, Polyendocrinopathy, Enteropathy, X-Linked (IPEX) in humans and scurfy in mice. Stem cell transplantation is the only cure for IPEX patients. The study of this rare disease has provided important insight into the mechanisms of immunosuppression, autoimmunity and tolerance and future studies may lead to novel strategies to treat not only patients with IPEX, but also those suffering from autoimmunity, graft-versus-host disease or cancer. PMID- 17917021 TI - A brief history of HLA. PMID- 17917017 TI - Safety of IGIV therapy and infusion-related adverse events. AB - Patients with primary immunodeficiency diseases have been treated with intravenous gammaglobulin (IGIV) for over 20 years. Gamma globulins were first introduced as a therapeutic modality in 1952 by Robert A. Good, who injected gamma globulins by the intramuscular (IM) route to treat patients with X-linked agammaglobulinemia. Subsequently, the clinical indications for gamma globulins therapy expanded from patients with Bruton's disease and other primary immune deficiency disorders to include a variety of autoimmune and inflammatory diseases. This expansion in clinical indications paralleled two critical steps in the development of gamma globulins preparations. First, proteolytic enzyme treated immunoglobulins became available for intravenous (IV) administration; unlike earlier IM preparations, IV preparations could be administered in larger volumes, were less painful, rapidly achieved physiologic concentrations of serum IgG, and were similar to native immunoglobulins with respect to their distribution of IgG subclasses and their pharmacokinetic features. Newer, highly purified IGIVs also became available. Second, significant efforts have been made to enhance the viral safety of IGIVs. Manufacturers no longer depend on Cohn Oncley or cold ethanol treatment as the only viral elimination step; rigorous donor screening, plasma testing by sensitive nucleic acid testing, and additional viral inactivation steps in the manufacturing processes for IGIVs have been implemented. In spite of the fact that this therapy has been life-saving in patients with primary immunodeficiency diseases, a number of adverse events have been reported as a result of its use. This paper will review some of the more frequent and important adverse events associated with the administration of IGIV, and discuss the mechanisms by which these reactions may occur. Our approach for replacement therapy with IGIV of patients with primary immunodeficiency diseases will be described, and some of the concerns related to the availability of commercial IGIV for this important patient population will be presented. Availability of IGIV product has been of particular concern to the clinical immunologists who take care of patients with primary immune deficiency disorders since there are no alternative therapies for these patients. PMID- 17917023 TI - Long-term follow-up in patients with severe combined immunodeficiency treated by bone marrow transplantation. AB - Immune reconstitution was studied in 31 long-term surviving patients after bone marrow transplantation for severe combined immunodeficiency. Donors in 7 cases were HLA-identical and in 25 cases HLA-haploidentical family members, and in 13 of these latter cases cytoreductive conditioning had been used prior to transplantation. At a mean follow-up of 15 years after transplantation (range 10 to 22 years), T cell numbers and functions had remained stable and within normal limits in the majority of patients. Marked variability however was observed with regard to reconstitution of B cell immunity. Furthermore numbers of circulating naive CD4+ T cells were variable and markedly diminished in a substantial proportion of patients at recent evaluations. Normal B cell immunity and persistently normal naive T cell numbers were strongly correlated with the continued detection of donor type CD34+ precursor cells in the patients marrow, which were absent in non conditioned patients. These findings indicate that stable donor precursor cell engraftment in the marrow may be of relevance for complete and stable long-term immune reconstitution in transplanted SCID patients. PMID- 17917022 TI - Blood and marrow transplantation: a perspective from the University of Minnesota. AB - On the occasion of the first meeting of the Robert A. Good Immunology Society in June of 2006, I was asked to provide a perspective on the history and progress of the field of bone marrow transplantation. I was honored to provide this perspective at that meeting and subsequently in this manuscript. This review has a strong University of Minnesota bias, as Minneapolis is a place where important roots in this field were developed. Minnesota is also where I have spent my career in this field learning the excitement of laboratory research beginning as a medical student under Bob Good and Carlos Martinez in 1960, and clinical research in pediatrics under Bill Krivit and Mark Nesbit beginning in 1970. This review is dedicated to two of my recently deceased mentors: Bob Good was a pioneer in so many ways and a true giant in immunology and blood and marrow transplantation. Bill Krivit taught me a great deal about genetic diseases and the critical role of compassion and understanding patients and their families in dealing with fatal illness and new treatments such as bone marrow transplantation that are often risky and themselves may result in suffering and death. My affection for Bob Good and Bill Krivit is unending. PMID- 17917024 TI - Hematopoietic stem cell transplantation across major genetic barriers. AB - The first successful demonstration that effective T cell depletion can enable immune reconstitution without causing graft versus host disease (GVHD) was achieved in 1980 using lectin-separated hematopoietic stem cells. In leukemia patients undergoing supralethal radio- and chemotherapy, T cell-depleted transplants are vigorously rejected by residual host T cells; this barrier was first overcome in 1993 by the use of megadose stem cell transplants. This clinical observation can be explained, in part, by the demonstration that cells within the CD34 compartments, as well as their immediate early myeloid progeny, are endowed with veto activity. Engraftment of mismatched hematopoietic stem cells following reduced intensity conditioning, still represents a major challenge. Progress has been made recently by using anti-3rd party veto CTLs and T regulatory cells. PMID- 17917025 TI - Matched unrelated bone marrow transplant for severe combined immunodeficiency. AB - Severe combined immunodeficiency (SCID) is a lethal disease unless allogeneic bone marrow transplantation (BMT), preferably from a family related HLA identical donor (RID) is given. Previously, some patients received HLA-mismatched related donors (MMRD) BMT, which often resulted in slow immune reconstitution and variable survival. Alternatively, HLA-matched unrelated donors (MUD) BMT have been suggested. Recently, we have directly compared outcome of patients with SCID who received either MMRD or MUD BMT. Survival after MUD BMT was significantly better than after MMRD BMT. Patients who received MUD BMT also had better engraftment of donor cells and immune reconstitution. Recent reports from other centers confirm these results finding that MUD BMT provides excellent survival and better immune reconstitution for patients with SCID. In conclusion, MUD BMT appears vastly superior to MMRD BMT and should be offered as first choice of treatment for patients with SCID when RID is unavailable. PMID- 17917026 TI - Immunological lessons learnt from patients transplanted with fully mismatched stem cells. AB - Fully HLA-mismatched stem cells from human fetal livers were transplanted into 17 infants and two fetuses to treat severe combined immunodeficiency disease in 1976 2000. Donor cell engraftment and immunological reconstitution were obtained in 14/19 patients, three of whom have been extensively and repeatedly studied immunologically during prolonged follow-up. T-cells were derived totally from donor cells; B-cells and antigen-presenting cells (APC) remained mainly of host origin. Due to class I and II mismatches between T-cells and all other cells (APC, B-cells, virus-infected target cells), limitations in the defense against infections in vivo and in T-cell functions in vitro (helper and cytotoxic activities) were predicted; however, these did not occur. Anti-tetanus toxoid responses (including specific antibody production) developed despite HLA disparities between T-cells and B-cells or APC in the chimeric children. Similarly, cytotoxic T-cells (of donor HLA phenotype) recognized host Epstein Barr virus-infected target cells. Recognition of antigenic peptide by T-cells under these conditions involved presentation by host allogeneic HLA molecules and not by self HLA antigens. Tolerance to donor antigens was acquired by clonal deletion; tolerance to host antigens existed despite the presence of many host reactive T-cells and involved clonal anergy. PMID- 17917030 TI - Innovative BMT methods for intractable diseases. AB - We have recently established new bone marrow transplantation (BMT) methods for the treatment of intractable diseases. The methods include the perfusion method (PM) for the collection of bone marrow cells, and intra-bone marrow (IBM)-BMT for the direct injection of collected whole bone marrow cells into the bone marrow cavity. The PM, in comparison with the conventional aspiration method, can minimize the contamination of bone marrow cells (BMCs) with T cells from the peripheral blood. Therefore, without removing T cells, no graft-versus-host disease (GvHD) develops in the case of the PM. Since BMCs collected by the PM contain not only hemopoietic stem cells (HSCs) but also mesenchymal stem cells (MSCs), the injection of both cells directly into the bone marrow cavity (IBM BMT) facilitates the engraftment of donor hemopoietic cells. In organ allografts with IBM-BMT, no graft failure occurs even if the radiation dose is reduced. In addition, IBM-BMT is applicable to regeneration therapy and various age associated diseases such as osteoporosis, since it can efficiently recruit donor derived normal MSCs. Finally, we show that IBM-BMT in conjunction with donor lymphocyte infusion (DLI) can prevent GvHD but suppress tumor growth. We believe that this strategy heralds a revolution in the field of transplantation (BMT and organ allografts) and regeneration therapy. PMID- 17917029 TI - Adoptive transfer of antigen-specific T-cells of donor type for immunotherapy of viral infections following allogeneic hematopoietic cell transplants. AB - Allogeneic marrow and cytokine-mobilized peripheral blood stem cells adequately depleted of T cells prevent acute and chronic forms of graft versus host disease in HLA-matched and non-identical hosts without any posttransplant immunosuppressive prophylaxis. Current cytoreductive regimens secure consistent durable engraftment, and full donor chimerism. The risk of relapse following such transplants in patients with AML and ALL has been low, and not different from that recorded following unmodified transplants. However, in HLA-disparate hosts the risk of infections caused by EBV, CMV, and certain fungi are increased. To address this limitation, others and we are exploring adoptive immunotherapies with in vitro generated, pathogen-specific T cells. Early clinical trials already indicate the potential of such T cells to treat and prevent life threatening diseases caused by these pathogens, particularly in recipients of T cell depleted grafts who do not require ongoing treatment with immunosuppressive agents, and therefore provide a permissive environment for the expansion and persistence of the T cells following adoptive transfer. New more predictable strategies are under development, which should allow such therapies to be broadly applicable. PMID- 17917028 TI - Chimerism and tetragametic chimerism in humans: implications in autoimmunity, allorecognition and tolerance. AB - The presence of cells or tissues from two individuals, chimeras, or the presence of cells and tissues that include the gonads, tetragametic chimerism can be detected by the analysis of cytogenetics and analysis of polymorphic genetic markers, using patterns of pedigree inheritance. These methodologies include determination of sex chromosomes, major histocompatibility complex (MHC) polymorphisms and panels of short tandem repeats (STRs) that include mitochondrial DNA markers. Studies routinely involve cases of temporal chimerism in blood transfusion, or following allotransplantation to measure the outcome of the organ, lymphopoietic tissues or bone marrow grafts. Demonstration of persistent chimerism is usually discovered in cases of inter-sexuality due to fusion of fraternal twins or in cases of fusion of embryos with demonstrable allogeneic monoclonality of blood which, excluded maternity or paternity when blood alone is used as the source of DNA. In single pregnancies it is possible to produce two kinds of microchimerism: feto-maternal and materno-fetal, but in cases of fraternal twin pregnancies it is possible to identify three different kinds which are related to cases of vanishing twins that can be identified during pregnancy by imaging procedures; (1) hematopoietic, (2) gonadal, and (3) freemartins when the twins have different sex and the individual born is a female with either gonadal or both gonadal and hematopoietic tissues. Fraternal twin pregnancies can also produce fusion of embryos. Such cases could be of different sex presenting with inter-sexuality or in same sex twins. One of such cases, the best studied, showed evidence of chimerism and tetragametism. In this regard, the case was studied because of disputed maternity of two of her three children. All tissues studied, except for the blood, demonstrated four genetic components but only two in her blood of four possible showed allogeneic monoclonality consistent with the interpretation that her blood originated from one hematopoietic stem cell. Also, microchimerism, due to traffic of cells via materno-fetal or feto maternal has been prompted by reports of their potential association with the development of autoimmune disorders including systemic lupus erythematosus (SLE) and systemic sclerosis, and in allotransplantation. In addition, their relevance of chimerism in the positive and negative selection of T cells in the thymus has not been addressed. T lymphocytes play a central role in controlling the acquired immune response and furthermore serve as crucial effector cells through antigen specific cytotoxic activity and the production of soluble mediators. Central tolerance is established by the repertoire selection of immature T lymphocytes in the thymus, avoiding the generation of autoreactive T cells. Expression of chimeric antigens in the thymus could modify the generation of specific T cell clones in chimeric subjects and these mechanisms could be important in the induction of central tolerance against foreign antigens important in allo transplantation. In this review, we discuss the genetics of chimerism and tetragametism and its potential role in thymic selection and the relevance in allotransplantation and autoimmune disorders. PMID- 17917031 TI - Growing organs for transplantation from embryonic precursor tissues. AB - Our recent data pinpoint a window of time in human and pig kidney organogenesis that may be optimal for transplantation into mature recipients. 'Window' transplants are defined by their remarkable ability to grow, differentiate and undergo vascularization, achieving successful organogenesis of urine-producing miniature kidneys. The transplanted tissue shows no evidence of trans differentiation into non-renal cell types or tumorogenicity, and displays reduced immunogenicity compared to its adult counterparts. Very recently, we demonstrated that this approach can be extended to transplantation of embryonic pig liver, pancreas, and lung tissue. Furthermore, it was demonstrated that E42 pancreatic tissue is optimally suited for induction of normoglycemia in diabetic mice. These results emphasize the importance of selecting precursors of the correct gestational age for optimal growth and function, and with reduced immunogenicity, and provide a proof of principle for the curative potential of E42 embryonic pig pancreatic tissue for transplantation in diabetic patients. PMID- 17917032 TI - What is the status of gene therapy for primary immunodeficiency? AB - The efforts to find satisfactory treatments for seriously ill patients with primary immunodeficiency have resulted in the development of important new therapeutic procedures with benefits reaching far beyond the relatively small number of patients affected with these rare disorders. Allogeneic bone marrow transplantation, immunoglobulin and enzyme replacement treatments and more recently gene therapy have all been introduced into clinical medicine as treatments for one or more of the primary immunodeficiency diseases. Beginning in 1990, gene-corrected T cells were first used to treat ADA deficiency SCID. With this demonstration that the gene-transfer procedure could be safely used to introduce functional transgenes into patient cells, clinical trials for a broad range of inherited disorders and cancer were started in the mid 90s. Of all these early clinical experiments, those addressing primary immunodeficiency have also been the most successful. Both ADA and X-SCID have now been cured using gene insertion into autologous bone marrow stem cells. In addition some patients with chronic granulomatous disease (CGD) have shown an unexpectedly high level of functionally corrected granulocytes in their blood following infusion of autologous gene-corrected bone marrow. There remain however a great many significant challenges to be overcome before gene therapy becomes the treatment of choice for these and other disorders. The use of genes as medicines is the most complex therapeutic system ever attempted and it may rake several more decades of work before its real potential as a treatment for both inherited and sporadic disorders if finally realized. PMID- 17917037 TI - Immunoglobulin variable regions in molecules exhibiting characteristics of innate and adaptive immune receptors. AB - The antigen combining sites of immunoglobulin (Ig) and T cell antigen receptors (TCRs), which are present in all jawed vertebrates, consist of a paired variable (V) domain heterodimer that exhibits varying degrees of germline- and extraordinarily high levels of somatically-derived variation. The near limitless variation in receptor specificity on the surface of individual lymphocytes is the basis for clonal selection in the adaptive immune response. A basic question arises as to whether or not there are other forms of immune-type receptors in vertebrates as well as in invertebrates that derive immune specificity through sequence differences in V domains. Our laboratory has discovered two such families of molecules, the novel immune-type receptors and the variable region containing chitin-binding proteins. Both families of molecules encode V domains that share some characteristics of adaptive immune receptors but likely mediate innate functions. PMID- 17917039 TI - Th1 or Th2 balance regulated by interaction between dendritic cells and NKT cells. AB - If Th1 or Th2 polarization could be artificially manipulated, effective immune responses would be generated depending on nature of the targets. In this study we attempted to regulate CD40 expressions on dendritic cells (DCs) in order to modify the T cell response. It was found that reducing agents selectively inhibited surface expression of CD40 on DCs. This finding may provide a new strategy of DC-mediated modulation of the Th1/Th2 balance. It was also shown that NKT-produced Th1/Th2 cytokine balance was under control of negative feedback loop through DCs. Th1 cytokine-pretreated DCs mainly induced Th2 cytokine production, whereas Th2 cytokine-pretreated DCs induced Th1 cytokine production by alpha galactosylceramide-stimulated NKT cells. The negative feedback regulation system could be applicable to therapeutics of various diseases based on immunological disorders. PMID- 17917040 TI - Impact of lymphocyte apoptosis on the innate immune stages of infection. AB - Infection of mice with Listeria monocytogenes has led to a puzzling observation: mice deficient in lymphocytes are more resistant during the early innate immune response. This is counterintuitive, because mice deficient in the adaptive immune response are unable to clear the infection and eventually die. This work will highlight some of our recent work on Listeria induced apoptosis and its immunological consequences. We show that Listeria produces a toxin, listeriolysin O (LLO), which causes apoptosis of lymphocytes in vitro and in vivo. Early during the infection, type I interferon sensitizes lymphocytes to die by LLO-induced apoptosis. The cell death peaks during the first two days of the infection, leading to the production of IL-10 and downregulation of anti-microbial activity in the spleen. The induction of apoptosis by Listeria creates an infective niche in the peri-arteriolar lymphoid sheaths of the spleen which allows the bacteria to grow exponentially. PMID- 17917038 TI - Immunity and protection, the unfolding of a tale. AB - Immunological memory is at the core of protective mechanisms against microbial pathogens and possibly of defenses against tumors. Here, a new perspective is offered on the qualitative and quantitative aspects of the T cell response as it relates to protection. Two main points are proposed. First, the conditions of the initial immune response (priming) are critically important in the induction of T cell memory and protection. Second, at the present time, protection against microbial pathogens appears to correlate with the function of central memory T cells. A series of considerations and suggestions are being made for new ways to optimize the induction of protective T cell responses by vaccination both in the immunologically naive and experienced individual; emphasis is placed on: dose of antigen, the availability of T cell help, avoidance of overt inflammatory conditions and efforts to decelerate cellular senescence in responding T cells. PMID- 17917041 TI - Interferon gamma, IL-12, IL-12R and STAT-1 immunodeficiency diseases: disorders of the interface of innate and adaptive immunity. AB - Susceptibility to mycobacterial infection has long been associated with defects in T cell immunity, such as those conferred by HIV infection or iatrogenic immune suppression. However, despite these well-recognized predispositions to clinical disease with tuberculosis and nontuberculous mycobacteria, the genetic disorders that are relatively specific for mycobacterial infection with nontuberculous bacteria and bacille Calmette Guerin (BCG) involve the innate immune pathways, and all engage interferon gamma and IL-12 production, signaling, and availability. PMID- 17917042 TI - Inherited human IRAK-4 deficiency: an update. AB - Interleukin-1 receptor-associated kinase-4 (IRAK-4), a member of the IRAK family, plays an essential role in Toll-like receptor- and IL-1 receptor-mediated signaling. We briefly review inherited human IRAK-4 deficiency, a recently described primary immunodeficiency leading to recurrent, invasive, pyogenic bacteria infection, and invasive pneumococcal disease in particular. PMID- 17917048 TI - Robert A Good and Ludwig van Beethoven: immortal beloveds: summary of the Robert A Good Immunology Society: Perspective in Immunology 2006 June 9-12 2006 St. Pete Beach, Florida. PMID- 17917046 TI - Human autoimmune diseases are specific antigen-driven T-cell diseases: identification of the antigens. AB - We are investigating the hypothesis that most human autoimmune diseases are specific antigen-driven T-cell diseases. T-cell clones responding to specific antigenic epitopes are responsible for the initiation and/or the propagation of these diseases. Similarly, specific antigen-driven T-cell responses are responsible for the rejection of organ allografts and the immune response to tumors. Activated T cells provide the "engine" for the chronic inflammation that is associated with autoimmune diseases, organ graft rejection and tumor immunity. The best way to identify whether specific antigen-driven T cell responses are involved in the initiation and/or propagation of these disorders is to investigate whether T cells that infiltrate relevant tissues from these diseases contain monoclonal or oligoclonal, that is to say clonally expanded, populations of T cells. Identification of the T-cell antigen receptor (TCR) transcripts employed by the clonally expanded T cells in these patients permits the identification of the specific antigens that elicit these T-cell responses. These antigens may be responsible for the pathogenesis of these diseases. We will summarize here certain of our findings in this area of research. PMID- 17917047 TI - Role of common gamma chain utilizing cytokines for immune reconstitution in HIV infection. AB - Many cytokines that utilize the common gamma (Cgamma) chain signaling pathway, viz Interleukin (IL)-2, IL-15, and IL-7 are known to be important for inducing T cell maturation, proliferation, or survival. Untreated chronic HIV infection is associated with profound quantitative and qualitative deficiency of CD4 T cells, which is partially reversed following highly active antiretroviral therapy (HAART). A subset of patients, however, fail to recover CD4 T cells despite virologic suppression. The role of Cgamma chain cytokines in influencing immune reconstitution following potent antiretroviral therapy is discussed. Maturation markers (naive, central memory, effector memory, and effector), cytokine receptors IL-2Rbeta, Cgamma chain, IL-7Ralpha, IL-15Ralpha, and cytokine-induced proliferative responses of T cells in a cohort of HIV-infected pediatric patients and adults classified on the basis of immunologic and virologic response to antiretroviral therapy were examined. The studies indicated that patients had increased percentages of effector memory CD8+ T cells in comparison to healthy volunteers. While patients with partially controlled viremia and poor CD4 T cell reconstitution manifested poor proliferative responses to anti-CD3 or HIV gag antigen stimulation, proliferative responses to Cgamma chain utilizing cytokines IL-2, IL-7, and IL-15 were robust. Another Cgamma chain utilizing cytokine, IL-21 had no influence on cellular proliferation but enhanced perforin expression in effector CD8 T cells. Thus, cytokine receptor deficiencies may contribute to immune deficiency in HIV-infected patients, and Cgamma chain cytokines may play an important role in vivo in immune homeostasis in lymphopenic patients by maintaining the memory subsets of T cells in patients with CD4 T cell deficiency. PMID- 17917049 TI - X-linked hypogammaglobulinemia and isolated growth hormone deficiency: an update. AB - X-linked hypogammaglobulinemia and isolated growth hormone deficiency (XLH-GHD, OMIM # 307200) is a primary immunodeficiency disorder characterized by pan hypogammaglobulinemia and isolated growth hormone deficiency. The disease, which is only known to occur in a single family, shares many features with X-linked agammaglobulinemia (XLA, OMIM # 300300). The current review summarizes the clinical, laboratory and genetic features of the disease as they have unfolded over the past quarter century since its description. PMID- 17917050 TI - Immunology at the University of Iowa. PMID- 17917051 TI - Structural requirements and applications of inhibitory oligodeoxyribonucleotides. AB - Synthetic oligodeoxyribonucleotides (ODN) bearing certain sequence characteristics mimic bacterial DNA by activating B cells and dendritic cells through Toll-like receptor (TLR) 9, an event that potentiates both humoral and cell-mediated immunity. ODN sharing some of the sequence characteristics of strong stimulatory (ST-) ODN, but substituting GGG for CGTT, competitively inhibit ST-ODN-driven events. An ODN with the same length and base composition as a strong ST-ODN, but lacking both ST- and IN-sequence requirements, has neither ST- nor IN-activity. Whereas, certain sequence changes strongly influence ST-ODN activity in human cells relative to mouse cells and B cells relative to non B cells, the strongest IN-ODN appear to work well in both species and multiple cell types. Converting from the natural phosphodiester backbone to a nuclease resistant phosphorothioate backbone increases the sensitivity to ST-ODN about 2 logs and to IN-ODN 3 logs, while increasing the impact of critical base changes in ST-ODN and diminishing it in IN-ODN. Examples where IN-ODN have been used in vivo to interrupt autoimmune and other TLR-9-induced inflammatory states are described. PMID- 17917052 TI - Modulation of NK cell activity by CpG oligodeoxynucleotides. AB - Oligodeoxynucleotides (ODN) with hypomethylated CpG motifs have been found to be potent stimulators of various aspects of innate and adaptive immunity. One of their major effects is the activation of natural killer (NK) killing activity in vitro and in vivo. There are several categories of CpG classified as type A, type B, and type C, although another category with inhibitory activity is being characterized further. CpG type A (CpG-A) is the most potent at activating NK cells. Examination of the cells and soluble mediators involved in this activation has led to an understanding of an interesting cascade of events. It appears that CpG activates dendritic cells (DC) which in turn activate NK-cells. This is not surprising since NK-cells do not seem to express TLR9, the CpG receptor. Of the various cytokines involved in NK-cell activation, it appears that type 1 interferon plays a pivotal role. Having activated NK-cells, DC themselves appear to become susceptible to lysis by the NK-cells they activated but with a delayed time kinetic. CpG ODN have been examined as monotherapeutic agents in murine tumor models. In one model, B16 melanoma, CpG ODN were very effective and NK cells were both necessary and sufficient for that effect. In another model, EL4 lymphoma, NK cells were necessary but not sufficient. Moreover, CpG were able to induce long-term survival in mice with established tumor. Studies in humans show similar results with potent activation in vitro. In a limited Phase I dose escalation study it also appeared that CpG ODN induce NK cell activation in humans in vivo. PMID- 17917053 TI - Multiple roles of TRAF3 signaling in lymphocyte function. AB - Members of the tumor necrosis factor receptor (TNFR) superfamily employ cytoplasmic adapter proteins called TNF-R associated factors (TRAF) to initiate and regulate signaling pathways. Although many of these receptors associate with TRAF3, it has been unclear how this TRAF functions in immune responses. New information appearing through the use of novel experimental models reveals that TRAF3 can mediate both activating and inhibitory signals, and can participate in regulation of multiple members of the TNFR superfamily. TRAF3 is also important for signaling via innate immune receptors, as well as an oncogenic mimic of a normal receptor that is implicated in promoting both malignancies and autoimmunity. PMID- 17917055 TI - The role of the T-cell costimulatory molecule Tim-1 in the immune response. AB - Upon encountering antigen, CD4+ T-cells become activated and can differentiate into subsets with distinct functional characteristics. One of these subsets is the Th2 cell, which generates large amounts of interleukin (IL)-4, -5, -10 and 13 in response to a subsequent encounter with antigen. In the context of a protective immune response, Th2 cells promote immune cell activation, antibody production, and inflammatory responses that help clear infections. However, aberrant responses by Th2 cells can lead to debilitating allergic diseases such as asthma. Thus, a reasonable approach toward gaining novel insights into immunity and allergic disease is to define the mechanisms that control Th2 cell differentiation and mature Th2 cell function. Recent work suggests that a protein called Tim-1 (T-cell immunoglobulin and mucin protein-1) is expressed on CD4+ T cells and plays a central role in regulating Th2 responses. Genetic analysis has linked polymorphisms in the human TIM1 gene to susceptibility to allergic disease, while studies involving mice have shown that ligation of Tim-1 promotes CD4+ T-cell activation. The signal transduction pathways downstream of Tim-1 are a relatively unexplored area. Continued study will undoubtedly reveal novel insights regarding the relationships between Tim-1, Th2 responses, and allergic disease. PMID- 17917054 TI - The isolator piglet: a model for studying the development of adaptive immunity. AB - The period from late gestation to weaning in neonatal mammals is a critical window when the adaptive immune system develops and replaces the protection temporarily provided by passive immunity and pre-adaptive antibodies. It is also when oral tolerance to dietary antigen and the distinction between commensal and pathogenic gut bacteria becomes established resulting in immune homeostasis. The reproductive biology of swine provides a unique model for distinguishing the effects of different factors on immune development during this critical period because all extrinsic factors are controlled by the experimenter. This chapter reviews this early stage of development and the usefulness of the piglet model for understanding events during this transitional stage. The review also describes the major features of the porcine immune system and the immune stimulatory and dysregulatory factors that act during this period. The value of the model to medical science in such areas as food allergy, organ transplantation, cystic fibrosis and the production of humanized antibodies for immuno-therapy is discussed. PMID- 17917056 TI - Natural Tregs, CD4+CD25+ inhibitory hybridomas, and their cell contact dependent suppression. AB - The natural CD4+CD25+ T regulatory (Treg) lymphocyte has emerged as a critical cell for controlling immune responses to self, foreign proteins, and pathogens. Identified initially by the constitutive expression of CD4 and CD25, natural Tregs suppress a variety of immune cells and responses, including CD4+CD25- proliferation and IL-2 production, and CD8 cell proliferation, IFNgamma production and CTL activity. Although natural Tregs require activation with specific antigen to attain their suppressive phenotype, once activated they execute inhibition in an antigen specific as well as non-specific (bystander) fashion. Treg suppression depends on IL-2, CD25, and cell:cell contact. The use of live cell imaging in vivo and in vitro to visualize the dynamic cell:cell interactions involving natural Tregs as well as the CD4+CD25+ Treg inhibitory hybridoma RD6 has refined the mechanistic models of contact dependent Treg suppression. PMID- 17917057 TI - Neutrophils and TRAIL: insights into BCG immunotherapy for bladder cancer. AB - Bladder cancer is a huge economic burden on the healthcare system and is responsible for approximately 5% of all cancer deaths in humans. Mycobacterium bovis BCG-based therapy is the treatment of choice for superficial bladder cancer. Bacillus Calmette-Guerin (BCG) instillation in the bladder results in a massive local inflammatory response that has secondary antitumor properties. Recent studies have demonstrated that neutrophils present in the bladder after BCG instillation release large amounts of the apoptosis-inducing molecule TRAIL, as well as chemokines that recruit other immune cells, suggesting that neutrophils play a key role in the antitumor response to BCG therapy. This review discusses the impact of these findings on the understanding of the antitumor mechanisms underlying BCG-based immunotherapy for bladder cancer. PMID- 17917058 TI - CD8 T cell memory development: CD4 T cell help is appreciated. AB - An important goal of vaccination strategies is to elicit long term, effective immunity. Therefore it is imperative to define the parameters that regulate the development and preservation of the numbers and functional quality of cells that confer this property to the host. CD8 T cells are a key component of the host adaptive immune response that helps eradicate invading viruses and other cell associated pathogens. Once the primary infection is controlled, the CD8 T cells transition from being effector cells into memory cells that act as sentinels of the immune system capable of rapidly purging the host of recurrent infections by the same pathogen. The factors that regulate and orchestrate this transition from effector CD8 T cells into functionally robust memory CD8 T cells are poorly understood. In recent years it has been determined that CD4 T cells play a vital role in the survival and functional responsiveness of memory CD8 T cells. However, the mechanism(s) of this interaction are still unclear. PMID- 17917059 TI - Roles of TRAF6 in CD40 signaling. AB - CD40 provides signals crucial to the activation of antigen-presenting cells during humoral and cell-mediated immune responses. A complex cohort of proteins interacts with the cytoplasmic domain of CD40 and mediates signaling. One member of this cohort is TNF receptor associated factor six (TRAF6). TRAF6 contributes to the CD40-mediated activation of NF-kappaB, stress-activated protein kinases, and perhaps other signaling molecules. TRAF6 may have roles as an adapter molecule, an activator of mitogen-activated protein kinases, and as a repressor of certain signaling circuits. Establishing the significance and interplay of these roles will lead to a more complete understanding of mechanisms important to the CD40-mediated activation of the immune system and will reveal novel targets for the development of therapeutic agents. PMID- 17917060 TI - The regulation of dendritic cell function by calcium-signaling and its inhibition by microbial pathogens. AB - Dendritic cells (DC) are the sentinels of the immune system, linking innate with adaptive responses. The functional responses of DC are subject to complex regulation and serve as targets for pathogens. Ca2+-mediated signal transduction pathways serve a central regulatory role in DC responses to diverse antigens, including TLR ligands, intact bacteria, and microbial toxins. This review summarizes the major mechanisms of Ca2+-signaling that DC utilize to regulate maturation and antigen presentation, including a Ca2+-calmodulin (CaM)-CaM kinase II pathway that is localized to phagosomes and is targeted by the human intracellular pathogen, Mycobacterium tuberculosis. Restoration of functional Ca2+ signaling in DC may provide a novel mechanism to enhance therapy and promote vaccine efficacy to infectious diseases, including tuberculosis. PMID- 17917061 TI - Respiratory dendritic cells: mediators of tolerance and immunity. AB - The respiratory tract is under constant bombardment from both innocuous and pathogenic material. The decision of how to respond to these challenges is mediated by a specialized set of antigen presenting cells within the lungs called dendritic cells (DC). Proper respiratory homeostasis requires that these respiratory DC (rDC) utilize both the local lung inflammatory environment as well as recognition of pathogen-specific patterns to determine whether to maintain homeostasis by either driving tolerance or immunity to the inhaled material. This review will focus on rDC and highlight how rDC regulate tolerance and immunity. PMID- 17917062 TI - Transcriptional control of complement receptor gene expression. AB - Immune complement is a critical system in the immune response and protection of host cells from damage by complement is critical during inflammation. The expression of the receptors for the inflammatory anaphylatoxin molecules is also key in immunity. In order to fully appreciate the biology of complement, a basic understanding of the molecular regulation of complement receptor gene expression is critical, yet these kinds of studies are lacking for many genes. Importantly, recent genetic studies have demonstrated that promoter-enhancer polymorphisms can contribute to pathology in diseases such as atypical hemolytic uremic syndrome. This review will focus on what is currently known about the genetic regulation of key protective complement receptors genes including CR1 (CD35), CR2 (CD21), Crry, MCP (CD46), DAF (CD55), and CD59. In addition, the regulation of the anaphylatoxin receptors genes, C3aR and C5aR (CD88) will also be discussed. Since new research continuously uncovers novel functions for these proteins, a greater appreciation of the mechanisms involved in gene regulation will be critical for understanding the biology of these molecules. PMID- 17917063 TI - Pathogenesis of acute and chronic central nervous system infection with variants of mouse hepatitis virus, strain JHM. AB - Infection of mice with variants of mouse hepatitis virus, strain JHM (MHV-JHM), provide models of acute and chronic viral infection of the central nervous system (CNS). Through targeted recombination and reverse genetic manipulation, studies of infection with MHV-JHM variants have identified phenotypic differences and examined the effects of these differences on viral pathogenesis and anti-viral host immune responses. Studies employing recombinant viruses with a modified spike (S) glycoprotein of MHV-JHM have identified the S gene as a major determinant of neurovirulence. However, the association of S gene variation and neurovirulence with host ability to generate anti-viral CD8 T cell responses is not completely clear. Partially protective anti-viral immune responses may result in persistent infection and chronic demyelinating disease characterized by myelin removal from axons of the CNS and associated with dense macrophage/microglial infiltration. Demyelinating disease during MHV-JHM infection is immune-mediated, as mice that lack T lymphocytes fail to develop disease despite succumbing to encephalitis with high levels of infectious virus in the CNS. However, the presence of T lymphocytes or anti-viral antibody can induce disease in infected immunodeficient mice. The mechanisms by which these immune effectors induce demyelination share an ability to activate and recruit macrophages and microglia, thus increasing the putative role of these cells in myelin destruction. PMID- 17917064 TI - Cytokine signaling to the cell cycle. AB - The proliferation of many myeloid and lymphoid cell populations is directly controlled by cytokine growth factors acting through a related family of cytokine receptors. This regulation implies that the signaling pathways activated by cytokine receptors must communicate with mechanisms that control mammalian cell cycle progression. Evidence for how these signaling pathways promote hematopoietic cell proliferation is considered along with their likely targets among the cell cycle regulators. PMID- 17917065 TI - The role of platelet CD154 in the modulation in adaptive immunity. AB - Platelets' primary role is hemostasis. However, a growing body of research has demonstrated that platelets are integral to the initiation of an inflammatory response and are potent effector cells of the innate immune response. Activated platelets express CD154, a molecule critical to adaptive immune responses, which has been implicated in platelet-mediated modulation of innate immune responses and inflammation. Recent studies utilizing CD154 knockout mice extend the role of platelet-derived CD154 to the modulation of adaptive immune response by enhancing antigen presentation, improving CD8+ T cell responses, and playing a critical function in T-dependent humoral immunity under physiological conditions. Together these data provide a basis for the expansion of the current paradigm of B cell activation and germinal center formation to include a role for platelets. PMID- 17917068 TI - Characterization of splenic CD21hi T2 B cells. AB - B cell development is a highly regulated process that initiates in the bone marrow (BM) of adult mice. After reaching the IgM+ immature stage in the BM, these B cells migrate to the spleen to complete maturation and incorporation into the long-lived peripheral lymphocyte pool. Studies have identified these splenic immature B cells, and have further attempted to delineate the sequence whereby they transition into mature B cells. As such, these immature splenic populations are termed transitional B cells and have been the focus of intense study. The review summarizes the phenotype and currently known functions of the four putative transitional B cell subsets identified to date. Although most appear to represent short-lived transitional B cells, the CD21hi T2 B cell population exhibits a number of qualities that question its label as a transitional B cell subset. PMID- 17917067 TI - Understanding respiratory syncytial virus (RSV) vaccine-enhanced disease. AB - Respiratory syncytial virus (RSV) is the most common cause of lower respiratory tract infection in infants and children worldwide. In addition, RSV causes serious disease in elderly and immune compromised individuals. RSV infection of children previously immunized with a formalin-inactivated (FI)-RSV vaccine is associated with enhanced disease and pulmonary eosinophilia that is believed to be due to an exaggerated memory Th2 response. As a consequence, there is currently no licensed RSV vaccine and detailed studies directed towards prevention of vaccine-associated disease are a critical first step in the development of a safe and effective vaccine. The BALB/c mouse model of RSV infection faithfully mimics the human respiratory disease. Mice previously immunized with either FI-RSV or a recombinant vaccinia virus (vv) that expresses the attachment (G) glycoprotein exhibit extensive lung inflammation and injury, pulmonary eosinophilia, and enhanced disease following challenge RSV infection. CD4 T cells secreting Th2 cytokines are necessary for this response because their depletion eliminates eosinophilia. Intriguing recent studies have demonstrated that RSV-specific CD8 T cells can inhibit Th2-mediated pulmonary eosinophilia in vvG-primed mice by as yet unknown mechanisms. Information gained from the animal models will provide important information and novel approaches for the rational design of a safe and efficacious RSV vaccine. PMID- 17917066 TI - PIP3 pathway in regulatory T cells and autoimmunity. AB - Regulatory T cells (Tregs) play an important role in preventing both autoimmune and inflammatory diseases. Many recent studies have focused on defining the signal transduction pathways essential for the development and the function of Tregs. Increasing evidence suggest that T-cell receptor (TCR), interleukin-2 (IL 2) receptor (IL-2R), and co-stimulatory receptor signaling are important in the early development, peripheral homeostasis, and function of Tregs. The phosphoinositide-3 kinase (PI3K)-regulated pathway (PIP3 pathway) is one of the major signaling pathways activated upon TCR, IL-2R, and CD28 stimulation, leading to T-cell activation, proliferation, and cell survival. Activation of the PIP3 pathway is also negatively regulated by two phosphatidylinositol phosphatases SHIP and PTEN. Several mouse models deficient for the molecules involved in PIP3 pathway suggest that impairment of PIP3 signaling leads to dysregulation of immune responses and, in some cases, autoimmunity. This review will summarize the current understanding of the importance of the PIP3 pathway in T-cell signaling and the possible roles this pathway performs in the development and the function of Tregs. PMID- 17917071 TI - Monoclonal antibody mechanisms of action in cancer. AB - Monoclonal antibodies (mAbs) are used extensively as treatment for cancer. A variety of mechanisms are thought to play important roles in mediating the observed anti-tumor effects of mAb. These include signaling mediated by cross linking of surface antigen that leads to cell death, blocking an activation signal that is necessary for continued cell growth, antibody-dependent cellular cytotoxicity (ADCC), complement mediated cytotoxicity (CMC) and the ability of mAb to alter the cytokine milieu or enhance development of an active anti-tumor immune response. In this review the in vitro, animal model and clinical results supporting each of these varied mechanisms is summarized as is the potential for these mechanisms to interact with each other. PMID- 17917070 TI - Strategies for differentiating embryonic stem cells (ESC) into insulin-producing cells and development of non-invasive imaging techniques using bioluminescence. AB - Diabetes is a chronic autoimmune disease that affects 4-5% of the world's population. If the present trends continue, diabetes would soon become a major/leading health problem worldwide. Hence there is an urgent need to develop novel approaches for the treatment of diabetes. While transplantation of the pancreas or that of isolated pancreatic islets can lead to the cure of the disease in some patients, immunological complications and the chronic shortage of donors makes it impossible to adequately treat all patients. Interestingly, embryonic stem cells (ESC) have emerged as a possible source of pluripotent cells that can be coaxed into insulin-producing cells (IPCs) that can be used to treat diabetes. However, until appropriate protocols have been established, this new technology will be difficult to tap into. Our laboratory is interested in developing new strategies for harnessing the pluripotency of ESC and differentiating them into IPCs that are stable and will continue to produce insulin in vivo. A second aspect is the non-availability of non-invasive imaging protocols. We show here that transcriptionally targeted luciferase expression can be used successfully to non-invasively monitor the transplanted cells in vivo. PMID- 17917069 TI - Regulation of interactions of Gram-negative bacterial endotoxins with mammalian cells. AB - Host defense against many invading Gram-negative bacteria (GNB) depends on innate immune recognition of endotoxin (lipopolysaccharides, LPS), unique surface glycolipids of GNB. Host responses to endotoxin must be highly sensitive but self limited. In mammals, optimal sensitivity is achieved by ordered interactions of endotoxin with several different extracellular and cell surface proteins-the LPS binding protein (LBP), CD14, MD-2, and Toll-like receptor (TLR) 4-reflecting the requirement for specific protein-endotoxin and protein-protein interactions. This complex reaction pathway also provides many ways to attenuate endotoxin-driven inflammation and can explain how differences in endotoxin structure, either intrinsic among GNB or induced by metabolic remodeling, can alter host responsiveness and thus the outcome of host-GNB interactions. Major goals of our research are to better understand: (1) the structural bases of specific host endotoxin interactions; (2) functional diversity among host endotoxin-binding proteins; and (3) how the actions of various endotoxin-binding proteins are regulated to permit optimal host responses to GNB infection. In addition, the identification of a water-soluble endotoxin:MD-2 complex that, depending on the structure of endotoxin or MD-2, has potent TLR4 agonist or antagonist properties suggests novel pharmacologic approaches to immuno-modulation. PMID- 17917072 TI - Immunotherapy of asthma using CpG oligodeoxynucleotides. AB - Asthma and other atopic disorders have increased in prevalence and severity over the past three decades. Reduced risk of atopic disease associated with early life exposure to infections and microbes has raised the possibility that pathogen associated molecular patterns (PAMPs) may confer protection against allergic disorders, a concept that has been named the "Hygiene Hypothesis". This relationship is most likely mediated through the induction of specific patterns of anti-atopic immune responses that follow engagement of innate immune mechanisms. Bacterial DNA is one such immunostimulatory microbe-associated ligand, whose properties can be mimicked by oligodeoxynucleotides (ODN) containing unmethylated cytosine-guanine dinucleotides in specific base sequences (CpG motifs), motifs characteristic of prokaryotic DNA that have been suppressed in eukaryotic DNA. Based initially on observations that CpG ODN induced Th1-type patterns of immune responses, we proposed that CpG ODN might represent a novel therapeutic strategy for the prevention and treatment of atopic disorders. Current understanding suggests multiple mechanisms of action of CpG ODN, but our initial hypothesis has been supported by extensive studies demonstrating, in animal models, efficacy in both incipient and established atopic asthma. These preclinical studies are now being translated into clinical trials exploring this new approach to immunotherapy for atopic disease. PMID- 17917073 TI - Analysis of candidate genes at the IBGC1 locus associated with idiopathic basal ganglia calcification ("Fahr's disease"). AB - Basal ganglia calcification (striatopallidodentate calcifications) can be caused by several systemic and neurological disorders. Familial Idiopathic Basal Ganglia Calcification (IBGC, "Fahr's disease"), is characterized by basal ganglia and extrabasal ganglia calcifications, parkinsonism and neuropsychiatric symptoms. Because of an increased use of neuroimaging procedures, calcifications of the basal ganglia are visualized more often and precociously. In 1999, a major American family with IBGC was linked to a locus on chromosome 14q (IBGC1). Another small kindred, from Spain, has also been reported as possibly linked to this locus. Here we report the main findings of the first 30 candidate genes sequenced at the IBGC1 locus during the process of searching for a mutation responsible for familial IBGC. During the sequencing process, we identified a heterozygous nonsynonymous single nucleotide polymorphism (exon 20 of the MGEA6/c TAGE gene) shared by the affected and not present in the controls. This SNP was randomly screened in the general population (348 chromosomes) in a minor allele frequency to 0.0058 (two heterozygous among 174 subjects). Another variation in this gene, in the exon 9, was found in the Spanish family. However, this variation was extremely common in the general population. Functional and population studies are necessary to fully access the implications of the MGEA6 gene in familial IBGC, and a complete sequencing of the IBGC1 locus will be necessary to define a gene responsible for familial IBGC. PMID- 17917074 TI - The role of TNF-alpha and its receptors in the production of beta-1,4 galactosyltransferase I and V mRNAs by rat primary astrocytes. AB - Glycosylation is one of the most important post-translational modifications. It is clear that the single step of beta-1,4-galactosylation is performed by a family of beta-1, 4-galactosyltransferases (beta-1,4-GalTs), and that each member of this family may play a distinct role in different tissues and cells. beta-1,4 GalT I and V are involved in the biosynthesis of N-linked oligosaccharides. beta 1,4-GalT I and V mRNAs are present in control astrocytes and affected by TNF alpha and lipopolysaccharide (LPS) stimuli. In this study, we examined the regulatory mechanisms of tumor necrosis factor-alpha (TNF-alpha)-affected production of beta-1,4-GalT I and V mRNAs. We show here that cultured astrocytes express TNF-alpha receptor 1 (TNFR1) and increased slightly after exposure to LPS. TNF-alpha and TNFR2 are not detected in control astrocytes and upregulated significantly with LPS stimulation and that activation of these receptors by TNF alpha affects expressions of beta-1,4-GalT I and V mRNAs. In addition, we observed that not only exogenous TNF-alpha but also TNF-alpha produced by astrocytes affected beta-1,4-GalT I and V mRNAs production in astrocytes. These results suggest that an autocrine loop involving TNF-alpha contributes to the production of beta-1,4-GalT I and V mRNA in response to inflammation. PMID- 17917075 TI - VEGF immunopositivity related to malignancy degree, proliferative activity and angiogenesis in ENU-induced gliomas. AB - Growth of solid tumors is highly dependent on angiogenesis. During tumor development, neoplastic cells switch to an angiogenic phenotype, playing a significant role in the expression of the vascular endothelial growth factor (VEGF). Seventy-two brain gliomas were induced in Sprague Dawley rats by prenatal exposure to ethylnitrosourea (ENU). Screening and location of tumors was carried out using magnetic resonance imaging (MRI). Conventional histology and immunocytochemistry for antibodies against glial fibrillary acidic protein (GFAP), S-100, NF, oligodendrocyte Ab-2, Ki-67, and VEGF165 were performed. The proliferation index (PI) was calculated from the Ki-67 labeling index, and the concentration of VEGF165 was quantified by enzyme-linked immunosorbent assay (ELISA). In vivo identification of macro- and microtumor appears to be useful to lead morphological and biochemical studies. Histopathology allows us to identify microtumors as classic oligodendrogliomas (CO; mean PI of 6.01 +/- 2.8%) and macrotumors as anaplastic oligodendrogliomas (AO; mean PI of 14.06 +/- 5%). Classic oligodendrogliomas show scarce VEGF165 expression whereas anaplastic ones display VEGF165 protein level 100-fold increased respect to CO. Astrocytes, neoplastic, and endothelial cells show differential immunostaining patterns from the border to the core of neoplasm. Positive structures for VEGF and their distribution vary according to PI increase. Anaplastic gliomas displaying VEGF positive intratumor capillaries correspond to the highest PI values. To identify the "angiogenic switch," we propose the glioma stage characterized by VEGF immunopositive neoplastic cells inside the tumor and positive endothelial cells surrounding it. PMID- 17917076 TI - A genetic variant in cytoskeleton motors amplifies susceptibility to leukoaraiosis in hypertensive smokers: gene-environmental interactions behind vascular white matter demyelinization. AB - One of the most frequent causes of an age-associated cognitive decline is the vascular white matter demyelinization of the brain referred to as leukoaraiosis (LA). The wide range of severity of the cognitive decline caused by LA can have numerous deleterious effects on the quality of life, leading overall to far reaching public health problems. Besides clinical risk factors such as hypertension and advanced age, genetic susceptibility factors are presumed to be of great importance in the development of LA. The protein kinesin, which is the main motor protein in the trafficking system of the mitochondria, can undergo functional damage under the circumstances of chronic hypoxia. This may give rise to a slowly developing metabolic crisis in the glia cells, a phenomenon hypothesized to account for the evolution of LA. Setting out from this assumption, we examined how the kinesin light-chain 1 (KNS2) G56836C single nucleotide polymorphism in intron 13 affects the susceptibility to LA. This genetic variant was found to be associated with cognitive disturbances and neurodegeneration, and it was presumed to affect the function of kinesin. The association analysis of the above genetic variant was performed in 229 patients with LA and 264 neuroimaging alteration-free controls. The KNS2 56836CC variant increased the risk of LA 7.76-fold in hypertensive smokers as compared with those not carrying this variant. This finding may be useful in everyday clinical practice by indicating the need for stricter preventive measures in CC carriers. PMID- 17917077 TI - Stable expression of a novel fusion peptide of thioredoxin-1 and ABAD-inhibiting peptide protects PC12 cells from intracellular amyloid-beta. AB - Amyloid-beta peptide-binding alcohol dehydrogenase (ABAD) inhibiting peptide, as a specific inhibitor between ABAD and amyloid-beta (Abeta), has been demonstrated to effectively inhibit Abeta peptide cytotoxicity. However, a major drawback is its short half-life, which results in the need for multiple applications and high synthesis costs. To overcome this, we established a lentiviral expression system that allowed the stable expression of the small ABAD-inhibiting peptide by fusion with cytosolic thioredoxin-1 (TRX). The fusion peptide, TA aptamer, was observed within PC12 cytoplasm and maintained both Abeta-binding ability and antioxygenic property similar to TRX. Our data showed that overexpression of both TRX and TA aptamer could protect PC12 cells from intracellular Abeta cytotoxicity. The present study suggests that TRX, as a cytosolic protein and a fusion motif, could not only assist ABAD-inhibiting peptide expression, cytoplasmic localization, but rebalance the disturbed "redox equilibrium" caused by intracellular Abeta in PC12 cells. PMID- 17917078 TI - Chronic mild stressors and diet affect gene expression differently in male and female rats. AB - While depression is reportedly more prevalent in women than men, a neurobiological basis for this difference has not been documented. Chronic mild stress (CMS) is a widely recognized animal model, which uses mild and unpredictable environmental stressors to induce depression. Studies of chronic stress, mainly in males, have reported an increase in the relative intake of "comfort food" as a means of counteracting the effects of stress. This study was designed to test the hypothesis that genes for certain neurotrophic factors, stress markers, and appetite regulators would be expressed differentially in male and female rats exposed to chronic, mild stressors with access to a preferred diet. Gene expression for neuropeptide Y was upregulated in females purely in response to stressors, whereas that for the epidermal growth factor receptor (EGFR) and arginine vasopressin (AVP) in males and fatty acid synthase (FASN) in females responded primarily to diet. Genes for brain-derived neurotrophic factor (BDNF), AVP, and the cocaine-amphetamine regulator of transcription (CART) in males, and leptin in females, showed a significant response to the interaction between stressors and diet. Every affected gene showed a different pattern of expression in males and females. This study confirms the intimate relationship between dietary intake and response to stress at the molecular level, and emphasizes the sex- and gene-specific nature of those interactions. Therefore, it supports a neurobiological basis for differences in the affective state response to stress in males and females. PMID- 17917080 TI - High salt culture conditions inhibit serum- and NGF- but not PMA-induced Egr-1 gene transcription in rat C6 glioma cells. AB - Recent studies have suggested that glial cells may play a physiologically important role in the retention and restoration of neuronal cell integrity, proposing the possibility that the proliferation and/or differentiation of glial cells may be related to pathological changes in neural functions in neurodegenerative diseases, and hence, it seems interesting to investigate the expression of genes related to the proliferation and differentiation of glial cells. Following this basic concept, we have previously examined the influence of culture conditions on egr-1 gene expression in rat C6 glioma cells and have shown that brief exposure of these cells to high salt culture medium can induce the down-regulation of egr-1 gene expression. In contrast, the long-term culture of these cells in high salt medium has been shown to primarily reduce their proliferation and secondarily elevate egr-1 gene transcription as a consequence of arresting the cell-cycle progression. Therefore, the effect of high salt culture medium on egr-1 gene expression seems practically unconfirmed, and remains to be further investigated. Then, the effects of various egr-1 gene inducers, such as serum, NGF and phorbol ester PMA, on Egr-1 mRNA levels in the glioma cells were examined under the high salt culture conditions. The brief exposure to high salt culture medium inhibited the elevation of Egr-1 mRNA levels induced by serum replenishment and NGF, but not induced by PMA. These results suggest that the suppression of serum action on egr-1 gene transcription may be the primary and essential event leading to the down-regulation of egr-1 gene expression under the high salt culture conditions. PMID- 17917079 TI - Molecular pathways associated with stress resilience and drug resistance in the chronic mild stress rat model of depression: a gene expression study. AB - The current antidepressant drugs are ineffective in 30 to 40% of the treated patients; hence, the pathophysiology of the disease needs to be further elucidated. We used the chronic mild stress (CMS) paradigm to induce anhedonia, a core symptom of major depression, in rats. A fraction of the animals exposed to CMS is resistant to the development of anhedonia; they are CMS resilient. In the CMS-sensitive animals, the induced anhedonic state is reversed in 50% of the animals when treating with escitalopram, whereas the remaining animals are treatment resistant. We used the microarray and the real-time quantitative reverse transcription polymerase chain reaction technique, as well as the ingenuity pathway analysis software to identify the differential gene expression pathways, which are associated with the occurrence of the treatment resistance and the stress-resilient rats. In the hippocampus, we found a significant upregulation of apoptotic pathways in the treatment-resistant animals and significantly increased expression levels of genes involved in hippocampal signaling in the CMS-resilient rats. We hypothesize that sensitivity to the stress-induced anhedonia in rats is correlated with the impairment of hippocampal neurogenesis. PMID- 17917082 TI - Tissue levels of adiponectin in breast cancer patients. AB - BACKGROUND: Adiponectin is a new adipocyte-secreted protein and associated with insulin-resistant status, such as type 2 diabetes mellitus and obesity. The inverse correlation between serum adiponectin levels and breast cancer risk was recently documented. On the other hand, the association of tissue adiponectin levels with breast cancer has not been previously reported. Thus, in the present study, the relationship between tissue adiponectin levels and breast cancer was evaluated. METHODS: We analyzed the correlation between tissue adiponectin levels and the occurrence of breast cancer in a case-control study comprising 27 women with diagnosed and histologically confirmed breast cancer and 33 women with fibroadenoma. In addition, the association of tissue adiponectin levels with the various classical risk factors, such as body mass index, menopausal status and, tumor size, stage, lymph node status, hormonal status were also studied. RESULTS: Tissue adiponectin levels in patients with breast cancer (0.75 +/- 0.06) were significantly higher than those in controls (0.68 +/- 0.1) (P = 0.02). The high tissue adiponectin levels were associated with significantly (P = 0.001) an increased risk for breast cancer compared with those in the low tissue adiponectin levels (OR, 1.34; 95% CI, 1.12-1.84) in breast cancer patients. In addition, postmenopausal women with the high tissue adiponectin levels showed a significantly (P = 0.003) an increased risk for breast cancer compared with women in low tissue adiponectin levels (OR, 1.63; 95% CI, 1.23-1.90). The correlation between BMI and breast cancer was not found (P > 0.05). Furthermore, the status of estrogen receptor, progesterone receptor, HER-2/neu receptor and lymph nodes involvement were established, no effect on the tissue adiponectin levels in patients with breast cancer and no correlations were detected among tumor stage, tumor size and the levels of tissue adiponectin (P > 0.05). CONCLUSION: Our results suggest that the high tissue adiponectin levels significantly detected in breast cancer patients and associated with an increased risk for breast cancer. PMID- 17917084 TI - Combination chemotherapy employing bispecific antisense oligonucleotides having binding sites directed against an autocrine regulated growth pathway and bcl-2 for the treatment of prostate tumors. AB - In previous studies we demonstrated that antisense oligonucleotides (oligos) against transforming growth factor-alpha (TGF-alpha [MR1]), its binding site the epidermal growth factor receptor (EGFR [MR2]), and the anti-apoptosis protein bcl 2 (MR4) are efficacious against prostate tumors. In recent reports we also describe how two of these mRNA directed binding sites can be synthesized sequentially within a single linear complementary strand and administered either in the presence or absence of additional therapeutic agents. In these continuing experiments "bispecific" oligo pairs were further evaluated in the presence or absence of Cytoxan, Taxol, or DES. One oligo pair recognized the binding sites for TGF-alpha and EGFR mRNA (TGF-alpha/EGFR [MR12] and EGFR/TGF-alpha [MR21]); another pair recognized binding sites for EGFR and bcl-2 (EGFR/bcl-2 [MR24] and bcl-2/EGFR [MR42]). Oligo pairs differ in their linear 5' to 3' binding site orientations, and were tested in vitro against PC-3 and LNCaP prostate tumor cell lines. Following cell attachment, incubations were for 2 days with the agents followed by 2 days in their absence. When tested against PC-3 cells and combined with LD50 Cytoxan, MR2, MR4, MR24, MR42 significantly inhibited 47.3, 45.7, 68.3, and 64.9%; with LD50 Taxol MR2, MR4, MR24, MR42 significantly inhibited 49.8, 45.8, 64.1, and 59.2%; and with LD50 DES MR2, MR4, MR24, MR42 significantly inhibited 66.6, 67.6, 64.3, and 67.2% respectively. Each agent significantly increased the inhibition produced by either oligo alone.LNCaP cells were also incubated with mono- and bispecific oligos in either the presence or absence of chemotherapeutics. MR2, MR4, MR24, MR42 produced significant inhibitions of 57.4, 58.4, 69.4, and 68.6% with LD50 Cytoxan; 70.4, 70.1, 73.6, and 74.0% with LD50 Taxol; and 49.8, 50.1, 59.6, and 53.9%, respectively with LD50 DES.A complete PC 3 experiment compared MR1, MR2, MR4, MR12, MR21, MR24 and MR42, in the presence of LD50 Cytoxan. Each oligo combined with Cytoxan significantly inhibited: MR1 by 51.0, MR2 by 55.0, MR4 by 58.0; MR12 by 56.0; MR21 by 61.1, MR24 by 65.5 and MR42 by 66.0%. Bispecifics directed against two different pathways, MR24, and MR42 were the most effective.A complete LNCaP experiment compared the same series of oligos also in the presence of LD50 Cytoxan. Each oligo combined with Cytoxan significantly inhibited: MR1 by 49.0, MR2 by 50.0, MR4 by 53.0; MR12 by 52.0; MR21 by 58.6, MR24 by 53.9 and MR42 by 58.0%. PMID- 17917083 TI - A case-control study of non-alcoholic fatty liver disease in breast cancer. AB - BACKGROUND: Patients with breast cancer sometimes present with increased liver enzymes during follow-up period that may be consistent with hepatic steatosis. This effect known as non-alcoholic fatty liver disease may be associated with the malignancy itself, drugs or some other well-known risk factors that may induce steatosis. We studied the influences of primary disease and treatment on steatosis in patients with breast cancer. MATERIALS AND METHODS: There were four groups of patients in our study. Group 1: 40 newly diagnosed, previously untreated breast cancer; Group 2: 45 cases of breast cancer treated with systemic therapy; Group 3: 40 cases of ovarian cancer; Group 4: 40 healthy women. Hepatic steatosis was evaluated by sonography by two radiologist, independently. We also evaluated major risk factors, biochemical findings, and influences of treatment on hepatic steatosis. RESULTS: We detected steatosis in 63%, 72%, 77%, and 48% of patients in groups 1, 2, 3, and 4, respectively. There was a statistically significant difference only between groups 3 and 4 (P = 0.045). However, grade 2 and 3 steatosis were more frequent in breast cancer patients (group 1 and 2), compared with mild steatosis in ovarian cancer patients and healthy women. Although a good correlation was found between tamoxifen use and chemotherapy on development of non-alcoholic fatty liver disease, no association of hepatic steatosis with transaminase levels was found, which might be of help for earlier detection of steatosis. AST/ALT ratio was found to have no impact on the rate of hepatic steatosis, contrary to the literature. CONCLUSION: Hepatic steatosis, excluding patients with grade 1 steatosis, which may be a normal variant, were more readily detected in patients with breast cancer. This effect was aggravated by use of tamoxifen, but not the chemotherapy. Non-alcoholic fatty liver disease in patients with breast cancer may be associated with the primary tumor itself or some well-known risk factors such as obesity, hyperlipidemia, and diabetes mellitus, which needs to be explored. PMID- 17917085 TI - The results of hypofractionated radiotherapy in 31 patients with high-grade gliomas. AB - In this prospective study, we investigated the effects of hypofractionated radiotherapy for patients with high-grade gliomas. About 31 patients with glioblastoma multiforme or anaplastic astrocytoma were studied between October 2003 and December 2004. Hypofractionated radiotherapy (3 Gy/fraction/day) was delivered to a total dose of 45 Gy in 15 fractions in 10 patients (32%) who had total excision before radiotherapy and to a total dose of 54 Gy in 18 fractions in 21 patients (68%) who had subtotal excision or biopsy alone. Sex, age, type of surgery, tumor grade, Karnofsky performance status, time between surgery and initiation of radiotherapy, and total radiotherapy dose were analyzed as potential prognostic factors for survival using the univariate log-rank method. The median follow-up was 15 months (4-16 months). A total of 15 patients (48%) died of their illness; 16 patients (52%) were still alive at the last follow-up. The median survival time was 8 months. Actuarial 1-year overall survival was 40%. Type of surgery, timing of radiotherapy after surgery, and initial Karnofsky performance status were significant prognostic factors for survival. No grade 3-4 acute or late neurotoxicity was observed. The tolerance of patients to hypofractionated RT was not different from that for conventional radiotherapy. This treatment schedule can be used for patients with high-grade gliomas. Future investigations are needed to determine the optimal fractionation for high-grade gliomas. PMID- 17917086 TI - Adjuvant chemoradiation for patients with adenocarcinoma of the pancreas: an expirence of single institute. AB - Only a small percentage of patients with pancreatic cancer have limited disease suitable for curative resection. Even with surgery, patients often have poor long term survival due to relapse of the disease. There are controversies about the adjuvant treatment of these patients. We reported the survival of resected pancreatic cancer from a single institute. About 128 consecutive patients who had complete resection of the pancreatic ductal adenocarcinoma were evaluated, retrospectively. Chemoradiotherapy (45 Gy plus 5-fluorouracil) was given to 63 patients. Fifty-five patients declined to take chemoradiotherapy or with poor performance status were observed without additional treatment. Eight patients took only chemotherapy and two patients took only radiotherapy. The median survival of chemoradiotherapy group was significantly higher than the observation group (13 months vs. 4 months, respectively; P < 0.001). In multivariate analyses the most important factors improving survival were the application of chemoradiation (P < 0.001), low-level serum LDH (P = 0.026), good performance status (P = 0.033) and low serum CA19-9 (P = 0.037). Although adjuvant chemoradiotherapy has a significant survival benefit when compared with the observation group, the survival data are still poor for pancreatic cancer. Therefore, we need more effective additional or adjuvant treatment modalities. PMID- 17917087 TI - Lack of prognostic value of EGFR mutations in primary resected non-small cell lung cancer. AB - PURPOSE: To investigate the prognostic value of epidermal growth factor receptor (EGFR) mutations in a series of Taiwanese patients with primary resected lung adenocarcinomas never treated with epidermal growth factor receptor tyrosine kinase (TK) inhibitor. PATIENTS AND METHODS: A total of 27 of 35 patients whose EGFR mutational status in exons 18, 19, and 21 of the TK domain had been previously determined using nested polymerase chain reaction (PCR) were included in this retrospective study. All 27 patients underwent potentially curative pulmonary resection. Clinicopathological information was obtained from patient records and pathology reports. Disease-free survival (DFS) and overall survival (OS) of patients were estimated with the Kaplan-Meier method, and Cox regression model was used for multivariate analysis. RESULTS: Heterozygous EGFR mutations were detected in 15 of 27 patients (55.5%). There was no significant difference in DFS between patients with wild-type EGFR (median, 16.87 months) and mutant EGFR (median, 18.13 months; P = 0.83). No significant difference in OS was also noted between the wild-type and mutant groups (P = 0.45, median follow-up 22.6 months). Cox regression model and multivariate analysis of survival difference by age, stage, histology, and adjuvant treatment did not reach statistical significance. CONCLUSION: EGFR mutations do not have significant prognostic value in primary resected non-small cell lung cancer (NSCLC), and further well-designed large prospective studies are warranted to determine the prognostic value of EGFR mutations in NSCLC. PMID- 17917088 TI - Inhibition of cell invasion and MMP production by a nutrient mixture in malignant liposarcoma cell line SW-872. AB - Liposarcoma, a malignancy of fat cells, is the most common soft tissue sarcoma. Though rare, poorly differentiated liposarcomas commonly metastasize to lungs and liver, leading to poor prognosis. Prevention of Extracellular matrix (ECM) degradation by inhibition of matrix metalloproteinases (MMPs) activity has been shown to be a promising therapeutic approach to inhibition of cancer progression. A nutrient mixture (NM) containing lysine, proline, ascorbic acid, and green tea extract has shown significant anticancer activity against a number of cancer cell lines. We investigated the effect of NM on liposarcoma cell line SW-872 proliferation (MTT assay), MMP secretion (gelatinase zymography), invasion through Matrigel, and apoptosis and morphology (live green caspase kit and H&E). Liposarcoma cell growth was inhibited by 36 and 61% at 500 and 1,000 microg/ml NM. Zymography demonstrated both MMP-2 and MMP-9 secretion, with PMA-enhanced MMP 9 activity. NM inhibited both MMPs with virtual total inhibition at 500 microg/ml NM. Invasion through Matrigel was inhibited at 100, 500, and 1,000 microg/ml by 44, 75, and 100%, respectively. Dose-dependent apoptosis of liposarcoma cells was evident with NM challenge, with virtually all cells exposed to 1,000 microg/ml NM in late apoptosis. H&E staining did not demonstrate any changes in morphology at lower concentrations. However, some apoptotic changes were evident at higher concentrations. In conclusion, NM significantly inhibited liposarcoma cell growth, MMP activity, and invasion and induced apoptosis in vitro-important parameters for cancer development, suggesting NM as a potential treatment strategy for liposarcoma. PMID- 17917089 TI - Phase I study of paclitaxel, carboplatin and UFT in chemo-naive patients with advanced non-small cell lung cancer (NSCLC). AB - OBJECTIVE: We conducted a phase I study of paclitaxel (PTX), carboplatin (CBDCA), and UFT in chemo-naive patients with advanced non-small cell lung cancer (NSCLC). METHOD: Twenty-one chemo-naive patients with advanced NSCLC were enrolled. The study was conducted as a phase I dose-escalation study of various doses of systemic PTX followed by CBDCA on day 1 and oral UFT (400 mg/m2) on days 1-5 and 8-12, with the cycle repeated at 21-day intervals. At least three patients were enrolled in each step. RESULTS: The main toxicities were neutropenia and paresthesia, but were tolerable and reversible in all cases. Overall response rate was 57% (12 out of 21). The MTD was not reached at the highest dose level after the first cycle. Given previous recommends of PTX at 225 mg/m2 and CBDCA AUC 6 for two-drug therapy, the recommended dose for the phase II study under our regimen was set at PTX 225 mg/m2 on day 1, CBDCA AUC 6 on day 1, and UFT 400 mg/m2 on days 1-5 and 8-12. CONCLUSION: The combination of PTX, CBDCA, and UFT showed promising activity and acceptable toxicity in these chemo-naive patients, supporting the development of this combination as a feasible chemotherapeutic option for advanced NSCLC. PMID- 17917090 TI - Phase II studies on docetaxel alone every third week, or weekly in combination with gemcitabine in patients with primary locally advanced, metastatic, or recurrent esophageal cancer. AB - BACKGROUND: The purpose of these studies was to compare efficacy and toxicity of docetaxel alone with the combination of gemcitabine and docetaxel for treatment of metastatic esophageal carcinoma. PATIENTS AND METHODS: These studies enrolled patients with histopathologically verified squamous cell carcinoma or adenocarcinoma of the esophagus or cardia. Between March 1997 and June 1999, 52 patients were enrolled in the initial Phase II study (Study 1). They were scheduled for treatment with docetaxel 100 mg/m2 every third week as a 1-h infusion. The second Phase II study between September 2000 and March 2003 included 65 patients (Study II). They were given docetaxel 30 mg/m2, administered as a 30-min i.v. infusion weekly for four times, followed by 2 weeks of rest, and gemcitabine starting with a dose of 750 mg/m2 (if well-tolerated 1,000 mg/m2) on days 1 and 15, followed by 3 weeks of rest. A new cycle began on day 36. Patients were premedicated with betamethasone 8 mg p.o. on the evening before, and 8 mg i.v. 30-60 min before the docetaxel infusion. Response was confirmed by computed tomography and assessed at 12 and 24 weeks. Toxicity was assessed according to WHO scales. RESULTS: In study I, 38 out of the 52 enrolled patients were valuable. Two patients experienced complete remission (CR) (5%), 10 patients partial remission (PR) (26%), nine patients stable disease (SD) (24%), and 17 patients showed progressive disease (PD) (45%). Toxicity mainly involved leukopenia, which in some cases required hospitalization and treatment with antibiotics. In Study II, 46 out of the 65 enrolled patients (70%) were assessable. Out of these, three patients (7%) had CR, eight patients (17%) had PR, 10 patients (22%) had SD, and 25 (54%) PD. Overall response was 24% while an additional 22% showed stable disease. Toxicity mainly consisted of leucopenia and pain. CONCLUSION: Docetaxel as a single agent is active in esophageal cancer, both in treatment naive and in previously treated patients with recurrent disease. The overall response rate was 31%, with a good-safety profile. The addition of gemcitabine is well tolerated, but adds no efficacy. Weekly administration of docetaxel may be less effective. It demonstrates moderate efficacy and the doses used provide an acceptable safety profile. PMID- 17917091 TI - Immunophenotypic profile and clinical characteristics in patients with advanced stage mantle cell lymphoma. AB - The objective of this study was to evaluate immunophenotypic profile along with clinical follow-up in patients with advanced stage mantle cell lymphoma (MCL), and their possible influence on overall survival (OS). Bone marrow (BM) cell and/or peripheral blood mononuclear cell flow cytometric analyses of the following antigens were performed: HLA-DR, CD19, CD20, CD22, CD23, CD25, CD10, SmIg, kappa, lambda, CD79b, CD38, FMC7, CD3, CD2, and CD5. There were 14 patients in IV CS, and 26 patients in CS V. All patients were treated with CHOP. Immunological markers showed a typical phenotype (CD5+ CD23-, Cyclin D1) in all cases. Pathohistological type of BM infiltration was predominantly diffuse (72.5%), and in remainder of patients, nodular. Comparison of patients with leukemic phase of MCL with CSIV (BM), has shown significantly higher expression of CD19, CD20, and CD23, followed by permanently negative expression of CD23. Patients with blastic variant of MCL had higher expression of CD23, compared to typical MCL (P < 0.001). Median OS was 20 months, and there were no significant OS-differences between CS IV and leukemic phase patients. Survival analyses showed that negative prognostic influence had high IPI (P < 0.01), presence of extranodal localization (P < 0.01), and diffuse type of BM involvement (P < 0.01). Using Cox regression according to OS, IPI had independent prognostic value (P < 0.001). Our results demonstrated that in the advanced MCL patients the most powerful prognostic factor was IPI, while extranodal localization and type of BM infiltration were of a limited value. PMID- 17917092 TI - The cell growth, morphology and immunocytochemistry of novel cell line established from a bone marrow of the patient with therapy-related myelodysplastic syndrome, entitled PC-MDS. AB - We report on cell growth, morphology, and immunocytochemistry of the first human cell line, PC-MDS, derived from a bone marrow of a patient with therapy-related myelodysplastic syndrome who had no overt leukemia post-MDS phase. This cell population consisted of fast-growing mononuclear cells. Standard cytochemistry methods for detection of MPO, lipids, glycogen and ANAE gave results as follows: MPO and SBB negative while PAS and ANAE positive. Positive cytochemical staining and immunophenotype analyses indicated that PC-MDS cells have some characteristics of the early myeloid precursor cell. As the first t-MDS derived cell line it could be a new tool in evaluation of complex biology of MDS and also serves as a model for diverse in-vitro research. PMID- 17917093 TI - Prognostic factors for malignant pericardial effusion treated by pericardial drainage in solid-malignancy patients. AB - PURPOSE: Malignant pericardial effusion is a frequent complication of advanced incurable malignancies and requires treatment. The purpose of this study was to identify prognostic factors for cytology-positive malignant pericardial effusion in patients treated by pericardial drainage. METHODS: We retrospectively analyzed a series of consecutive patients diagnosed with cytologically positive malignant pericardial effusion who were treated by pericardial drainage at the National Cancer Center Hospital, Tokyo. RESULTS: A total of 88 patients with pericardial effusion were treated by pericardial drainage, 60 patients were diagnosed with cytological positive malignant pericardial effusion including 32 with non-small cell lung cancer, 13 with breast cancer, 8 with gastrointestinal cancer, and 7 with miscellaneous cancers. Subxiphoid pericardiostomy was performed in 50 of the patients and percutaneous tube pericardiostomy in the other 10 patients. Malignant pericardial effusion recurred in 14 patients, and pericardial drainage was performed again in 9 of them. The median overall survival time was 6.1 months, and the 1-year survival rate was 28%. A multivariate analysis revealed the following significant negative prognostic factors: performance status, development of malignant pericardial effusion during chemotherapy, mediastinal lymph node enlargement, and cytologic type. (P = 0.03, 0.02, 0.01, 0.001, respectively). CONCLUSION: Patients with poor prognostic factors may be better to consider as indication of palliative therapy, even if oncologic emergency had been resolved rapidly by drainage. PMID- 17917094 TI - Capecitabine plus oxaliplatin (xelox) in the treatment of chemotherapy-naive patients with metastatic colorectal cancer. AB - BACKGROUND: Capecitabine and oxaliplatin are both synergistically active against metastatic colorectal cancer (MCRC). We evaluated our experience at two centers with capecitabine and oxaliplatin combination (XELOX) in previously untreated patients with MCRC. PATIENTS AND METHODS: We reviewed medical records of 85 previously untreated patients with MCRC who received first-line XELOX regimen. Oxaliplatin was given at a dose of 130 mg/m2 on day 1 in combination with capecitabine 1500 mg/m2/day on days 1-14 every 3 weeks. RESULTS: Seventy-six of 85 patients were evaluated for response and toxicity. Patients with a follow up of less than 6 months were excluded from the study. Objective response rate was 46% including 8 complete responses (10.5%) and 27 partial responses (35.5%). Additionally, 20 patients (26.3%) had disease stabilization at least 3 months after the treatment. The patients were followed for a median 12.5 months (range 2 32). Median time to disease progression (TTP) was 11 months (range 2-27 months). Median overall survival (OS) time has not yet been reached. One-year survival rate was 66%. Toxicity was modest with infrequent grade 3-4 adverse effects. CONCLUSION: XELOX is an active regimen against MCRC in the first-line setting with favorable toxicity profile. Our results appear to be comparable, if not superior, to the results of other reports of first-line XELOX therapy in respect to objective response rates, survival data, and safety profile. Convenience with oral administration of every 3-week schedule makes XELOX regimen a compelling therapeutic option in the treatment of first-line MCRC. PMID- 17917096 TI - Systemic lupus Erythematosus and IgA multiple myeloma: a rare association? AB - The coexistence of systemic lupus Erythematosus (SLE) and multiple myeloma (MM) is uncommon and the pathogenetic mechanisms underlying this association remain unclear. We report the case of a woman who was diagnosed with SLE in 1993 aged 57, then developing IgA lambda type MM in the IIB clinical stage 7 years later. The SLE was treated successfully with methylprednisolone and chloroquine, and low dose maintenance steroid was continued with bisphosphonate protection until December 1994 when she suffered multiple vertebral fractures. She continued to receive 4 mg alternate day methylprednisolone and calcitonin until she decided to discontinue her own treatment 2 years later. In 2000, while still in stable SLE remission, she was diagnosed with MM. Protein electrophoresis revealed the IgA lambda paraprotein (40.5 g/l) and she had a Bence Jones (BJ) proteinuria of the lambda light chain type. Bone marrow trephine biopsy revealed a massive patchy infiltrate of abnormal plasmocytes (70%), while an extensive x-ray skeletal survey did not show any new fractures or osteolysis. The patient was treated according to the VMCP protocol without attaining a plateau phase. There was a similar poor clinical response to second and third line treatments (VAD, Thalidomide, Melphalan, and high dose dexamethasone). After 4 years of refractory disease the patient died from severe bilateral pneumonia. This case is discussed with reference to the literature. PMID- 17917095 TI - Position-dependent expression of GADD45alpha in rat brain tumours. AB - Although the complex and multifactorial process of tumour growth has been extensively studied for decades, our understanding of the fundamental relationship between tumour growth dynamics and genetic expression profile remains incomplete. Recent studies of tumour dynamics indicate that gene expression in solid tumours would depend on the distance from the centre of the tumour. Since tumour proliferative activity is mainly localised to its external zone, and taking into account that generation and expansion of genetic mutations depend on the number of cell divisions, important differences in gene expression between central and peripheral sections of the same tumour are to be expected. Here, we have studied variations in the genetic expression profile between peripheral and internal samples of the same brain tumour. We have carried out microarray analysis of mRNA expression, and found a differential profile of genetic expression between the two cell subsets. In particular, one major nuclear protein that regulates cell responses to DNA-damaging and stress signals, GADD45alpha, was expressed at much lower levels in the peripheral zone, as compared to tumour core samples. These differences in GADD45alpha mRNA transcription levels have been confirmed by quantitative analysis via real time PCR, and protein levels of GADD45alpha also exhibit the same pattern of differential expression. Our findings suggest that GADD45alpha might play a major role in the regulation of brain tumour invasive potential. PMID- 17917097 TI - Malignant mesothelioma of the tunica vaginalis testis: a case report and review of the literature. AB - Most often, mesotheliomas involve the serosal membranes of the pleura and peritoneum. Sometimes, mesothelial proliferations are identified in other locations. A mesothelioma, within the tunica vaginalis of the paratesticular region is rare but often fatal malignancy of the male genitalia. Despite aggressive surgical and systemic therapy the prognosis remains poor with only rare long-term survivors. We report a case of malignant mesothelioma of the tunica vaginalis in 45-years-old and review of the literature is presented. PMID- 17917099 TI - Tumor lysis syndrome after treatment with gemcitabine for metastatic transitional cell carcinoma. AB - Tumor lysis syndrome is a set of life threatening complication that can arise from treatment of high tumor burden, drug sensitive, and rapidly proliferating neoplasm particularly of hematological origin. It is rarely described in patients with solid tumors. We report the first case of tumor lysis syndrome in a man with metastatic renal pelvic transitional cell carcinoma after gemcitabine treatment. Despite aggressive therapy, he died 2 weeks after TLS was diagnosed. Our experience demonstrates that administration of gemcitabine for metastatic renal pelvic transitional cell carcinoma may induce acute tumor lysis syndrome, which necessitates frequent laboratory monitoring and prompt initiation of appropriate therapeutic measures. PMID- 17917098 TI - Tamoxifen induced-thrombocytopenia: it does occur. AB - Tamoxifen has been in the center of management of hormone-sensitive breast cancer. Furthermore, it represents the best example of chemo-prevention: It reduces the incidence of invasive breast cancer. However, it has some side effects, several of them are severe. Oncologists, hematologists, internal medicine specialists and gynecologists must know this agent in detail. Authors report a very rare side effect, thrombocytopenia, and discuss it briefly. PMID- 17917100 TI - Lung, bone, skeletal muscles and cutaneous metastases from adenoid cystic carcinoma of the parotid gland: a case report and review of the literature. AB - Adenoid cystic carcinoma is the second most common malignancy of the major and minor salivary glands after mucoepidemoid carcinoma. The risk of distant metastases is approximately 20-50%. Although bone, the central nervous system and the other organs may become involved, the lungs are favored sites for metastases. Skeletal muscle and cutaneous metastases from adenoid cystic carcinoma of the parotid gland are extremely rare. In this case, a 40-year-old man with lung and bone metastases followed by skeletal muscle and cutaneous metastases from adenoid cystic carcinoma of the right parotid gland is presented. PMID- 17917101 TI - Solitary iris metastasis from breast cancer with dramatic course: case report. AB - Intraocular metastases are the most common malignancy of eye. Breast cancer is more frequently a cause of intraocular metastases. As a first metastatic site, iris and ciliary body are relatively rare. We report a case of a 52-year-old woman, operated for breast cancer 16 months ago and diagnosed multiple brain metastases 1 month ago. After first course of chemotherapy she was admitted to hospital with the complaints of eye pain and she recognized a solid mass on iris. Iris and ciliary metastases were diagnosed by ophthalmological examination. Because of the patient's poor general condition, diagnostic aspiration from eye metastasis could not be performed. Intramedullary mass was determined 1 month later and she died 2 months later. Ciliary body and iris metastases of breast cancer must be considered as a manifestation of aggressive clinical course and poor prognosis. The eye metastases of breast cancer are a part of systemic illness and must be treated by systemic chemotherapy. PMID- 17917102 TI - Two cases of sarcoidosis-lymphoma syndrome. AB - An association between sarcoidosis and lymphoproliferative diseases (LD), the sarcoidosis-lymphoma syndrome, has been previously described, and may be attributed to the underlying immunological abnormalities that occur during the sarcoidosis disease process. We report two patients who developed Hodgkin's disease and diffuse large B-cell non-Hodgkin's lymphoma (NHL) subsequent to their diagnosis of sarcoidosis after latency periods of 6 years and 18 years respectively. Both patients developed histologically-proven sarcoidosis late in life, at 46 years and 58 years, and had differing clinical courses. The first had radiographically staged II chronic progressive respiratory sarcoidosis (RS) and required long-term methotrexate to control the disease, while the second achieved a spontaneous remission of her stage I intrathoracic RS. After treatment, the patient with Hodgkin's disease remains in remission 2.5 years following six cycles of ABVD protocol chemotherapy and involved-field radiotherapy, while the NHL patient remains in remission at 3 years following six cycles of R-COP protocol chemotherapy. Clinicians should be aware of the potential risks of malignancy, and especially of LD in sarcoidosis patients. They should be alerted to the possibility of additional pathology by any atypical clinical features, and should biopsy new lesions and adenopathy to exclude any coexistent neoplasm. PMID- 17917104 TI - Tracking TrkA's trafficking: NGF receptor trafficking controls NGF receptor signaling. AB - Growth factors such as the neurotrophins promote neuronal survival and shape neuronal morphology. Neurotrophin receptors are located on the surface of axons and dendrites and must convey their signal retrogradely to the nucleus to influence transcription of target genes. The distance between the site of receptor activation and the nucleus is tremendous. How is the retrograde transmission of survival signals being achieved? Recent work showed that signaling endosomes containing neurotrophin receptors and associated downstream kinases undergo retrograde vesicular transport along microtubules, propelled by the molecular motor dynein. The next objective in the "neurotrophin receptor trafficking meets signal transduction field" will be to elucidate the traffic control mechanisms governing the directed movement of signaling endosomes. Much is already known on the trafficking of the receptor for epidermal growth factor, EGFR. We will summarize the known traffic control mechanisms for EGFR and hypothesize whether EGFR-relevant traffic control mechanisms might also be relevant for neurotrophin receptor traffic control. Moreover, we speculate about potential implications of neurotrophin receptor traffic jams for neurodegenerative diseases. PMID- 17917103 TI - Ionic channel function in action potential generation: current perspective. AB - Over 50 years ago, Hodgkin and Huxley laid down the foundations of our current understanding of ionic channels. An impressive progress has been made during the following years that culminated in the revelation of the details of potassium channel structure. Nevertheless, even today, we cannot separate well currents recorded in central mammalian neurons. Many modern concepts about the function of sodium and potassium currents are based on experiments performed in nonmammalian cells. The recent recognition of the fast delayed rectifier current indicates that we need to reevaluate the biophysical role of sodium and potassium currents. This review will consider high quality voltage clamp data obtained from the soma of central mammalian neurons in the view of our current knowledge about proteins forming ionic channels. Fast sodium currents and three types of outward potassium currents, the delayed rectifier, the subthreshold A-type, and the D-type potassium currents, are discussed here. An updated current classification with biophysical role of each current subtype is provided. This review shows that details of kinetics of both sodium and outward potassium currents differ significantly from the classical descriptions and these differences may be of functional significance. PMID- 17917106 TI - Searching for the ligands of odorant receptors. AB - Through the sense of smell mammals can detect and discriminate between a large variety of odorants present in the surrounding environment. Odorants bind to a large repertoire of odorant receptors located in the cilia of olfactory sensory neurons of the nose. Each olfactory neuron expresses one single type of odorant receptor, and neurons expressing the same type of receptor project their axons to one or a few glomeruli in the olfactory bulb, creating a map of odorant receptor inputs. The information is then passed on to other regions of the brain, leading to odorant perception. To understand how the olfactory system discriminates between odorants, it is necessary to determine the odorant specificities of individual odorant receptors. These studies are complicated by the extremely large size of the odorant receptor family and by the poor functional expression of these receptors in heterologous cells. This article provides an overview of the methods that are currently being used to investigate odorant receptor-ligand interactions. PMID- 17917105 TI - Modulation of ligand-gated ion channels by antidepressants and antipsychotics. AB - It is generally accepted that antidepressants and antipsychotics mediate their therapeutic effects via specific interaction with processes related to synaptic neurotransmission in the central nervous system. Besides their well-known classical mechanisms of action, antidepressants and antipsychotics show widely unknown effects, which might also contribute to the pharmacological profile of these agents. There is growing evidence that an interaction of these drugs with allosteric modulatory sites of ligand-gated ion channels (LGICs) might represent a yet unknown principle of action. Such interactions of psychopharmacological drugs with LGICs might play an important role both for the therapeutic efficacy and the side effect profile of these agents. In this review, we focus on the direct interaction of antidepressants and antipsychotics with LGICs, which may provide a basis for the development of novel psychopharmacological drugs. PMID- 17917107 TI - Hormonal regulation of clonal, immortalized hypothalamic neurons expressing neuropeptides involved in reproduction and feeding. AB - The hypothalamus has been particularly difficult to study at the molecular level because of the inherent cellular heterogeneity and complexity of neuronal circuits within. We have generated a large number of immortalized, clonal cell lines through retroviral gene transfer of the oncogene SV40 T-Ag into primary murine hypothalamic neuronal cell cultures. A number of these neuronal cell lines express neuropeptides linked to the control of feeding behavior and reproduction, including neuropeptide Y (NPY) and neurotensin (NT). We review recent studies on the direct regulation of NPY gene expression by estrogen, and the leptin-mediated control of signal transduction pathways and NT transcription. These studies provide new insights into the direct control of neuropeptide synthesis by hormones and nutrients at a mechanistic level in the individual neuron, not yet possible in the whole brain. Using these novel cell models, we expect to contribute substantially to the understanding of how individual neuronal cell types control overall endocrine function, especially with regard to two of the most well-known roles of distinct peptidergic neurons; these being the control of reproduction and energy homeostasis. PMID- 17917108 TI - Searching for a role of NCX/NCKX exchangers in neurodegeneration. AB - Control of intracellular calcium signaling is essential for neuronal development and function. Maintenance of Ca2+ homeostasis depends on the functioning of specific transport systems that remove calcium from the cytosol. Na+/Ca2+ exchange is the main calcium export mechanism across the plasma membrane that restores resting levels of calcium in neurons after stimulation. Two families of Na+/Ca2+ exchangers exist, one of which requires the co-transport of K+ and Ca2+ in exchange for Na+ ions. The malfunctioning of Na+/Ca2+ exchangers has been related to the development of pathological conditions in the regulation of neuronal death after hypoxia-anoxia, brain trauma, and nerve injury. In addition, the Na+/Ca2+ exchanger function has been associated with impaired Ca2+ homeostasis during aging of the brain, as well as with a role in Alzheimer's disease by regulating beta-amyloid toxicity. In this review, we summarize the current knowledge about the Na+/Ca2+ exchanger families and their implications in neurodegenerative disorders. PMID- 17917110 TI - Exercise-induced neuroprotection in SMA model mice: a means for determining new therapeutic strategies. AB - Due to the prevalence of neuromuscular disorders such as amyotrophic lateral sclerosis and spinal muscular atrophy in modern societies, defining new and efficient strategies for the treatment of these two neurodegenerative diseases has become a vital and still unfulfilled urge. Several lines of experimental evidence have emphasized the benefits of regular exercise training in mouse models for these affections in terms of life span increase and improvement of both motor capacities and motoneuron survival. Identifying molecules that could mimic the neuroprotective effects of exercise represents a promising way to find novel therapies. Some of the effects of exercise are caused by the overproduction of circulating neurotrophic factors, such as IGF-I, whereas others may be due to modifications of the intrinsic properties of the motoneurons within the spinal cord. The causal relationship that links these potential effects of exercise training and the improvement of motor capacity and life span expectancy is consequently discussed. PMID- 17917109 TI - Heat shock proteins and amateur chaperones in amyloid-Beta accumulation and clearance in Alzheimer's disease. AB - The pathologic lesions of Alzheimer's disease (AD) are characterized by accumulation of protein aggregates consisting of intracellular or extracellular misfolded proteins. The amyloid-beta (Abeta) protein accumulates extracellularly in senile plaques and cerebral amyloid angiopathy, whereas the hyperphosphorylated tau protein accumulates intracellularly as neurofibrillary tangles. "Professional chaperones", such as the heat shock protein family, have a function in the prevention of protein misfolding and subsequent aggregation. "Amateur" chaperones, such as apolipoproteins and heparan sulfate proteoglycans, bind amyloidogenic proteins and may affect their aggregation process. Professional and amateur chaperones not only colocalize with the pathological lesions of AD, but may also be involved in conformational changes of Abeta, and in the clearance of Abeta from the brain via phagocytosis or active transport across the blood-brain barrier. Thus, both professional and amateur chaperones may be involved in the aggregation, accumulation, persistence, and clearance of Abeta and tau and in other Abeta-associated reactions such as inflammation associated with AD lesions, and may, therefore, serve as potential targets for therapeutic intervention. PMID- 17917111 TI - Pain facilitation and activity-dependent plasticity in pain modulatory circuitry: role of BDNF-TrkB signaling and NMDA receptors. AB - Pain modulatory circuitry in the brainstem exhibits considerable synaptic plasticity. The increased peripheral neuronal barrage after injury activates spinal projection neurons that then activate multiple chemical mediators including glutamatergic neurons at the brainstem level, leading to an increased synaptic strength and facilitatory output. It is not surprising that a well established regulator of synaptic plasticity, brain-derived neurotrophic factor (BDNF), contributes to the mechanisms of descending pain facilitation. After tissue injury, BDNF and TrkB signaling in the brainstem circuitry is rapidly activated. Through the intracellular signaling cascade that involves phospholipase C, inositol trisphosphate, protein kinase C, and nonreceptor protein tyrosine kinases; N-methyl-D-aspartate (NMDA) receptors are phosphorylated, descending facilitatory drive is initiated, and behavioral hyperalgesia follows. The synaptic plasticity observed in the pain pathways shares much similarity with more extensively studied forms of synaptic plasticity such as long-term potentiation (LTP) and long-term depression (LTD), which typically express NMDA receptor dependency and regulation by trophic factors. However, LTP and LTD are experimental phenomena whose relationship to functional states of learning and memory has been difficult to prove. Although mechanisms of synaptic plasticity in pain pathways have typically not been related to LTP and LTD, pain pathways have an advantage as a model system for synaptic modifications as there are many well-established models of persistent pain with clear measures of the behavioral phenotype. Further studies will elucidate cellular and molecular mechanisms of pain sensitization and further our understanding of principles of central nervous system plasticity and responsiveness to environmental challenge. PMID- 17917112 TI - New molecular insights into cellular survival and stress responses: neuroprotective role of cellular prion protein (PrPC). AB - Knowledge of the physiological function of cellular prion protein has been acquired from prion diseases such as Creutzfeldt-Jakob disease, as well as PRNP knock out and transgenic mice. Recent progress in neurobiology has further delineated the neuroprotective role played by cellular prion protein. In this paper, we review the role of cellular prion protein in cell survival including its antiapoptotic effect on Bax-mediated cell death and its responses to various environmental stresses including oxidative stress, and ischemia. Finally, we discuss the significance of cellular prion protein in different neurodegenerative diseases and the possible development of future therapies. PMID- 17917113 TI - Roles of transient receptor potential vanilloid subtype 1 and cannabinoid type 1 receptors in the brain: neuroprotection versus neurotoxicity. AB - Transient receptor potential vanilloid subtype 1 (TRPV1), also known as vanilloid receptor 1 (VR1), is a nonselective cation channel that is activated by a variety of ligands, such as exogenous capsaicin (CAP) or endogenous anandamide (AEA), as well as products of lipoxygenases. Cannabinoid type 1 (CB1) receptor belongs to the G protein-coupled receptor superfamily and is activated by cannabinoids such as AEA and exogenous Delta-9-tetrahydrocannabinol (THC). TRPV1 and CB1 receptors are widely expressed in the brain and play many significant roles in various brain regions; however, the issue of whether TRPV1 or CB1 receptors mediate neuroprotection or neurotoxicity remains controversial. Furthermore, functional crosstalk between these two receptors has been recently reported. It is therefore timely to review current knowledge regarding the functions of these two receptors and to consider new directions of investigation on their roles in the brain. PMID- 17917114 TI - TWEAK and the central nervous system. AB - Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is a member of the tumor necrosis factor superfamily that acts on responsive cells via binding to a cell surface receptor named fibroblast growth factor-inducible 14 (Fn14). TWEAK can regulate numerous cellular responses in vitro and in vivo. Recent studies have indicated that TWEAK and Fn14 are expressed in the central nervous system (CNS), and that in response to a variety of stimuli, including cerebral ischemia, there is an increase in TWEAK and Fn14 expression in perivascular astrocytes, microglia, endothelial cells, and neurons with subsequent increase in the permeability of the blood-brain barrier (BBB) and cell death. Furthermore, there is a growing body of evidence indicating that TWEAK induces the activation of the NF-kappaB in the CNS with release of proinflammatory cytokines and matrix metalloproteinases. In addition, inhibition of TWEAK activity by either treatment with a Fn14-Fc fusion protein or neutralizing anti-TWEAK antibodies has shown therapeutic efficacy in animal models of ischemic stroke, cerebral edema, and multiple sclerosis. PMID- 17917115 TI - Function and dysfunction of CNG channels: insights from channelopathies and mouse models. AB - Channels directly gated by cyclic nucleotides (CNG channels) are important cellular switches that mediate influx of Na+ and Ca2+ in response to increases in the intracellular concentration of cAMP and cGMP. In photoreceptors and olfactory receptor neurons, these channels serve as final targets for cGMP and cAMP signaling pathways that are initiated by the absorption of photons and the binding of odorants, respectively. CNG channels have been also found in other types of neurons and in non-excitable cells. However, in most of these cells, the physiological role of CNG channels has yet to be determined. CNG channels have a complex heteromeric structure. The properties of individual subunits that assemble in specific stoichiometries to the native channels have been extensively investigated in heterologous expression systems. Recently, mutations in human CNG channel genes leading to inherited diseases (so-called channelopathies) have been functionally characterized. Moreover, mouse knockout models were generated to define the role of CNG channel proteins in vivo. In this review, we will summarize recent insights into the physiological and pathophysiological role of CNG channel proteins that have emerged from genetic studies in mice and humans. PMID- 17917116 TI - Morphogenic signaling in neurons via neurotransmitter receptors and small GTPases. AB - Several neurotransmitters including serotonin and glutamate have been shown to be involved in many aspects of neural development, such as neurite outgrowth, regulation of neuronal morphology, growth cone motility and dendritic spine shape and density, in addition to their well-established role in neuronal communication. This review focuses on recent advances in our understanding of the molecular mechanisms underlying neurotransmitter-induced changes in neuronal morphology. In the first part of the review, we introduce the roles of small GTPases of the Rho family in morphogenic signaling in neurons and discuss signaling pathways, which may link serotonin, operating as a soluble guidance factor, and the Rho GTPase machinery, controlling neuronal morphology and motility. In the second part of the review, we focus on glutamate-induced neuroplasticity and discuss the evidence on involvement of Rho and Ras GTPases in functional and structural synaptic plasticity triggered by the activation of glutamate receptors. PMID- 17917117 TI - Laminins in peripheral nerve development and muscular dystrophy. AB - Laminins are extracellular matrix (ECM) proteins that play an important role in cellular function and tissue morphogenesis. In the peripheral nervous system (PNS), laminins are expressed in Schwann cells and participate in their development. Mutations in laminin subunits expressed in the PNS and in skeleton muscle may cause peripheral neuropathies and muscular dystrophy in both humans and mice. Recent studies using gene knockout technology, such as cell-type specific gene targeting techniques, revealed that laminins and their receptors mediate Schwann cell and axon interactions. Schwann cells with disrupted laminin expression exhibit impaired proliferation and differentiation and also undergo apoptosis. In this review, we focus on the potential molecular mechanisms by which laminins participate in the development of Schwann cells. PMID- 17917120 TI - Are pheromones detected through the main olfactory epithelium? AB - A major sensory organ for the detection of pheromones by animals is the vomeronasal organ (VNO). Although pheromones control the behaviors of various species, the effect of pheromones on human behavior has been controversial because the VNO is not functional in adults. However, recent genetic, biochemical, and electrophysiological data suggest that some pheromone-based behaviors, including male sexual behavior in mice, are mediated through the main olfactory epithelium (MOE) and are coupled to the type 3 adenylyl cyclase (AC3) and a cyclic nucleotide-gated (CNG) ion channel. These recent discoveries suggest the provocative hypothesis that human pheromones may signal through the MOE. PMID- 17917119 TI - Myotubularin-related (MTMR) phospholipid phosphatase proteins in the peripheral nervous system. AB - Myotubularin-related proteins (MTMRs) constitute a broad family of ubiquitously expressed phosphatases with 14 members in humans, of which eight are catalytically active phosphatases, while six are catalytically inactive. Active MTMRs possess 3-phosphatase activity toward both PtdIns3P and PtdIns(3, 5)P 2 poliphosphoinositides (PPIn), suggesting an involvement in intracellular trafficking and membrane homeostasis. Among MTMRs, catalytically active MTMR2 and inactive MTMR13 have a nonredundant function in nerve. Loss of either MTMR2 or MTMR13 causes Charcot-Marie-Tooth type 4B1 and B2 neuropathy, respectively, characterized by demyelination and redundant loops of myelin known as myelin outfoldings. In Mtmr2-null mouse nerves, these aberrant foldings occur at 3-4 weeks after birth, a time when myelination is established, and Schwann cells are still elongating to reach the final internodal length. Moreover, Mtmr2-specific ablation in Schwann cells is both sufficient and necessary to provoke CMT4B1 with myelin outfoldings. MTMR2 phospholipid phosphatase might regulate intracellular trafficking events and membrane homeostasis in Schwann cells during postnatal nerve development. In this review, we will discuss recent findings on the MTMR family with a major focus on MTMR2 and MTMR13 and their putative role in Schwann cell biology. PMID- 17917118 TI - Group I metabotropic glutamate receptors: a role in neurodevelopmental disorders? AB - Group I metabotropic glutamate receptors (mGlu1 and mGlu5) are coupled to polyphosphoinositide hydrolysis and are involved in activity-dependent forms of synaptic plasticity, both during development and in the adult life. Group I mGlu receptors can also regulate proliferation, differentiation, and survival of neural stem/progenitor cells, which further support their role in brain development. An exaggerated response to activation of mGlu5 receptors may underlie synaptic dysfunction in Fragile X syndrome, the most common inherited form of mental retardation. In addition, group I mGlu receptors are overexpressed in dysplastic neurons of focal cortical dysplasia and hemimegaloencephaly, which are disorders of cortical development associated with chronic epilepsy. Drugs that block the activity of group I mGlu receptors (in particular, mGlu5 receptors) are potentially helpful for the treatment of Fragile X syndrome and perhaps other neurodevelopmental disorders. PMID- 17917121 TI - FXTAS in spanish patients with ataxia: support for female FMR1 premutation screening. AB - Fragile X-associated tremor/ataxia syndrome (FXTAS) is a newly described disorder characterized by progressive action tremor and ataxia that occurs in premutation carriers of the FMR1 gene. The incidence of FMR1 premutated carriers in the general population is relatively high, and therefore FXTAS might explain a considerable number of sporadic, late-onset ataxias. To better establish the prevalence of FXTAS among undiagnosed Spanish patients with ataxia, we have performed a FMR1 premutation screening. Our results evidenced three individuals carrying premutated alleles, giving an estimated FXTAS prevalence of 1.95% among patients with late-onset ataxia (1.15% for males and 3% for females). Molecular characterization of premutation carriers evidences lower fragile X mental retardation 1 protein levels and increased FMR1 mRNA levels. Clinical and neuroimaging findings support FXTAS diagnosis in these patients. Because of the high prevalence of FMR1 premutation in the general population, the description and characterization of the FXTAS syndrome is of great interest as it may represent one of the more common monogenic causes of ataxia, tremor, and dementia. The results obtained in this study demonstrate that FXTAS should be incorporated to spinocerebellar ataxia genetic screening protocols. Early diagnosis of these patients benefits not only them but also the rest of the family that should be advised for the fragile X syndrome. PMID- 17917124 TI - Value added by data sharing: long-term potentiation of neuroscience research. A commentary on the 2007 SfN Satellite Symposium on data sharing. PMID- 17917125 TI - Mapping the human brain: new insights from FMRI data sharing. AB - The sharing of primary data in the field of neuroscience has received considerable scrutiny from scientific societies and from science journals. Many see this as value added for science publishing that can enhance and inform secondary examination of data and results. Still others worry that data sharing is an undue burden for researchers with little long term value to science. But examples of how data sharing can be done successfully do exist. The fMRI Data Center, established at Dartmouth College in 2000 and now based at the University of California Santa Barbara, has worked to facilitate the open sharing of neuroimaging data from peer-reviewed papers to foster progress in cognitive science. The fMRI study on the representation of objects in the human occipital and temporal cortex, published in 2000 in the Journal of Cognitive Neuroscience (JOCN), marked the first deposition in the new database. Despite initial concerns about fMRI data sharing, this data set was frequently downloaded. We describe the original results of distributed brain activation patterns elicited by faces and objects in the human visual system, and overview several secondary analyses by independent investigators. A philosopher tested Husserl's temporal components of consciousness, whereas other brain imagers deployed new analytic tools, from Dynamic Causal Modeling, which estimates the neural interactions between cortical regions, to a novel method for constructing reproducibility maps. These re analyses revealed new findings not reported in the original study, provided new perspectives on visual perception, generated new predictions, and resulted in new collaborations and publications in high profile journals. PMID- 17917126 TI - Successes and rewards in sharing digital reconstructions of neuronal morphology. AB - The computer-assisted three-dimensional reconstruction of neuronal morphology is becoming an increasingly popular technique to quantify the arborization patterns of dendrites and axons. The resulting digital files are suitable for comprehensive morphometric analyses as well as for building anatomically realistic compartmental models of membrane biophysics and neuronal electrophysiology. The digital tracings acquired in a lab for a specific purpose can be often re-used by a different research group to address a completely unrelated scientific question, if the original investigators are willing to share the data. Since reconstructing neuronal morphology is a labor-intensive process, data sharing and re-analysis is particularly advantageous for the neuroscience and biomedical communities. Here we present numerous cases of "success stories" in which digital reconstructions of neuronal morphology were shared and re-used, leading to additional, independent discoveries and publications, and thus amplifying the impact of the "source" study for which the data set was first collected. In particular, we overview four main applications of this kind of data: comparative morphometric analyses, statistical estimation of potential synaptic connectivity, morphologically accurate electrophysiological simulations, and computational models of neuronal shape and development. PMID- 17917122 TI - Heterotrimeric G proteins and the single-transmembrane domain IGF-II/M6P receptor: functional interaction and relevance to cell signaling. AB - The G protein-coupled receptor (GPCR) family represents the largest and most versatile group of cell surface receptors. Classical GPCR signaling constitutes ligand binding to a seven-transmembrane domain receptor, receptor interaction with a heterotrimeric G protein, and the subsequent activation or inhibition of downstream intracellular effectors to mediate a cellular response. However, recent reports on direct, receptor-independent G protein activation, G protein independent signaling by GPCRs, and signaling of nonheptahelical receptors via trimeric G proteins have highlighted the intrinsic complexities of G protein signaling mechanisms. The insulin-like growth factor-II/mannose-6 phosphate (IGF II/M6P) receptor is a single-transmembrane glycoprotein whose principal function is the intracellular transport of lysosomal enzymes. In addition, the receptor also mediates some biological effects in response to IGF-II binding in both neuronal and nonneuronal systems. Multidisciplinary efforts to elucidate the intracellular signaling pathways that underlie these effects have generated data to suggest that the IGF-II/M6P receptor might mediate transmembrane signaling via a G protein-coupled mechanism. The purpose of this review is to outline the characteristics of traditional and nontraditional GPCRs, to relate the IGF-II/M6P receptor's structure with its role in G protein-coupled signaling and to summarize evidence gathered over the years regarding the putative signaling of the IGF-II/M6P receptor mediated by a G protein. PMID- 17917128 TI - Software development vis a vis collaboration in interdisciplinary research. PMID- 17917127 TI - Sharing and reusing gene expression profiling data in neuroscience. AB - As public availability of gene expression profiling data increases, it is natural to ask how these data can be used by neuroscientists. Here we review the public availability of high-throughput expression data in neuroscience and how it has been reused, and tools that have been developed to facilitate reuse. There is increasing interest in making expression data reuse a routine part of the neuroscience tool-kit, but there are a number of challenges. Data must become more readily available in public databases; efforts to encourage investigators to make data available are important, as is education on the benefits of public data release. Once released, data must be better-annotated. Techniques and tools for data reuse are also in need of improvement. Integration of expression profiling data with neuroscience-specific resources such as anatomical atlases will further increase the value of expression data. PMID- 17917129 TI - Clinical diagnosis based on bayesian classification of functional magnetic resonance data. AB - We describe a method for classifying subjects based on functional magnetic resonance (fMR) data, using a method combining a Bayesian-network classifier with inverse-tree structure (BNCIT), and ensemble learning. The central challenge is to generate a classifier from a small sample of high-dimensional data. The principal strengths of our method include the nonparametric multivariate Bayesian network representation, and joint performance of feature selection and classification. Preliminary results indicate that this method can detect regions characterizing group differences, and can, on the basis of activation levels in these regions, accurately classify new subjects. PMID- 17917130 TI - Automated axon tracking of 3D confocal laser scanning microscopy images using guided probabilistic region merging. AB - This paper presents a new algorithm for extracting the centerlines of the axons from a 3D data stack collected by a confocal laser scanning microscope. Recovery of neuronal structures from such datasets is critical for quantitatively addressing a range of neurobiological questions such as the manner in which the branching pattern of motor neurons change during synapse elimination. Unfortunately, the data acquired using fluorescence microscopy contains many imaging artifacts, such as blurry boundaries and non-uniform intensities of fluorescent radiation. This makes the centerline extraction difficult. We propose a robust segmentation method based on probabilistic region merging to extract the centerlines of individual axons with minimal user interaction. The 3D model of the extracted axon centerlines in three datasets is presented in this paper. The results are validated with the manual tracking results while the robustness of the algorithm is compared with the published repulsive snake algorithm. PMID- 17917131 TI - hESC adaptation, selection and stability. PMID- 17917133 TI - Hematopoietic stem cell aging: wrinkles in stem cell potential. AB - Hematopoietic stem cells (HSC) continuously replenish the blood and immune systems. Their activity must be sustained throughout life to support optimal immune responses. It has been thought that stem cells may be somewhat protected from age because of their perpetual requirement to replenish the blood, however studies over the past 10 years have revealed dramatic changes in HSC function and phenotype with respect to age. When the number of HSC within murine bone marrow is measured, an increase in concentration and absolute number of HSC within the bone marrow is observed as the animal ages, paralleled with increased homogeneity of stem cell marker expression. Results from transplantation studies demonstrate that although there is a decline in hematopoietic output on a per-cell basis, the increase in number provides sufficient, yet abnormal, blood production throughout the lifespan of the animal. HSC may play a role in immunosenescence through cell fate decisions leading to an overproduction of myeloid cells and an underproduction of lymphocytes. When examining gene expression of aged HSC, recent studies have highlighted several key factors contributing to increased inflammation, stress response and genomic instability. Here, we will review the general phenotype observed with aging of the hematopoietic system, focusing on the HSC, and compile recent expression profiling efforts that have examined HSC aging. PMID- 17917132 TI - Aging and the germ line: where mortality and immortality meet. AB - Germ cells are highly specialized cells that form gametes, and they are the only cells within an organism that contribute genes to offspring. Germline stem cells (GSCs) sustain gamete production, both oogenesis (egg production) and spermatogenesis (sperm production), in many organisms. Since the genetic information contained within germ cells is passed from generation to generation, the germ line is often referred to as immortal. Therefore, it is possible that germ cells possess unique strategies to protect and transmit the genetic information contained within them indefinitely. However, aging often leads to a dramatic decrease in gamete production and fecundity. In addition, single gene mutations affecting longevity often have a converse effect on reproduction. Recent studies examining age-related changes in GSC number and activity, as well as changes to the stem cell microenvironment, provide insights into the mechanisms underlying the observed reduction in gametogenesis over the lifetime of an organism. PMID- 17917134 TI - Aging, graying and loss of melanocyte stem cells. AB - Hair graying is one of the prototypical signs of human aging. Maintenance of hair pigmentation is dependent on the presence and functionality of melanocytes, neural crest derived cells which synthesize pigment for growing hair. The melanocytes, themselves, are maintained by a small number of stem cells which reside in the bulge region of the hair follicle. The recent characterization of the melanocyte lineage during aging has significantly accelerated our understanding of how age-related changes in the melanocyte stem cell compartment contribute to hair graying. This review will discuss our current understanding of hair graying, drawing on evidence from human and mouse studies, and consider the contribution of melanocyte stem cells to this process. Furthermore, using the melanocyte lineage as an example, it will discuss common theories of tissue and stem cell aging. PMID- 17917137 TI - Breast cancer risk assessment and genetic testing: complexities, conundrums, and community. PMID- 17917135 TI - Endothelial precursor cells. AB - The discovery and subsequent characterization of endothelial precursor cells (EPCs) has stimulated interest in their potential use in older persons. Understanding the mechanisms that underlie EPC availability and function has important clinical implications for this age group. In this review, we discuss aspects of EPCs that are relevant to their role in angiogenesis and cardiovascular disease. We then review the limited data on features of EPCs that are known to be altered in aging and might better define their clinical utility in older persons. PMID- 17917136 TI - Intrinsic changes and extrinsic influences of myogenic stem cell function during aging. AB - The myogenic stem cell (satellite cell) is almost solely responsible for the remarkable regeneration of adult skeletal muscle fibers after injury. The availability and the functionality of satellite cells are the determinants of efficient muscle regeneration. During aging, the efficiency of muscle regeneration declines, suggesting that the functionality of satellite cells and their progeny may be altered. Satellite cells do not sit in isolation but rather are surrounded by, and influenced by, many extrinsic factors within the muscle tissue that can alter their functionality. These factors likely change during aging and impart both reversible and irreversible changes to the satellite cells and on their proliferating progeny. In this review, we discuss the possible mechanisms of impaired muscle regeneration with respect to the biology of satellite cells. Future studies that enhance our understanding of the interactions between stem cells and the environment in which they reside will offer promise for therapeutic applications in age-related diseases. PMID- 17917139 TI - Susceptibility to breast cancer: hereditary syndromes and low penetrance genes. AB - Several genes are associated with hereditary susceptibility to breast cancer. Most notably these include BRCA1 and BRCA2; however, other less common gene mutations which confer elevated breast cancer risk are associated with Cowden syndrome, Li-Fraumeni syndrome, Peutz-Jeghers syndrome, ataxia-telangiectasia heterozygosity and hereditary diffuse gastric cancer. In this article we highlight the genetic epidemiology, gene function, genotype-phenotype correlations, cancer risks and clinicopathologic findings for the cancer susceptibility genes related to these syndromes. We also examine genes, such as CHEK2, which confer a lower penetrance for breast cancer in comparison to these highly penetrant genes. PMID- 17917138 TI - Assessing breast cancer risk and BRCA1/2 carrier probability. AB - By identifying individuals with an increased risk of breast cancer, health professionals can offer prevention strategies tailored to individual risk levels. Such strategies may include early initiation of cancer screening, more frequent screening, targeted therapeutic or behavioral interventions, or prophylactic surgery. In order to achieve clinical benefits with this approach, however, risk assessment strategies and effective prevention measures must be available. In this article we review current knowledge about cancer risk assessment for unaffected women and probability models for identifying individuals who are carriers of a mutation in BRCA1 or BRCA2, the two genes most commonly implicated in hereditary breast cancer. We review BRCA1 and BRCA2 mutations in various ethnic populations and how this information factors into risk assessment. Additionally, we summarize the current guidelines for when to make a referral to genetic services for risk assessment and evaluation. PMID- 17917140 TI - Management of women at increased risk for hereditary breast cancer. AB - This chapter reviews the management of women at increased risk for hereditary breast cancer. Screening strategies, emphasizing new data regarding magnetic resonance imaging (MRI), are discussed. It is recommended that women at risk for hereditary breast cancer consider annual MRI in addition to mammography and clinical breast examinations. Next, currently available data regarding chemoprevention and surgical prophylaxis for women at high risk for hereditary breast cancer are reviewed. Chemoprevention with tamoxifen reduces the risk of breast cancer by approximately 50% in a heterogeneous population of high risk women; however, the benefit in women at risk for hereditary breast cancer is less clear. In contrast, surgical prophylaxis with bilateral mastectomy reduces the risk of breast cancer by at least 90% and bilateral oophorectomy reduces the risk of breast cancer by approximately 50% among high risk women. Next, ways in which BRCA mutation-associated breast cancers differ from sporadic breast cancers are reviewed. Unique management issues pertaining to women with hereditary breast cancer are then discussed with an emphasis on surgical decision making and peri diagnostic genetic counseling and testing. Finally, the management of women at risk for hereditary breast cancer in whom deleterious mutations are not identified is reviewed. PMID- 17917141 TI - Genetic counseling for breast cancer risk: general concepts, challenging themes and future directions. AB - This article reviews the elements and process of genetic counseling for breast cancer risk, including contracting, informed consent, and psychosocial assessment and counseling. Case studies and pedigrees are utilized to illustrate current approaches to issues and challenges in the field. For example, the following topics are explored: test result interpretation, including uninformative BRCA1/2 test results; testing strategies and test selection; family concerns; patient follow-up and recontact; risk counseling in double heterozygotes; and reproductive options for mutation carriers. Concerns in specific populations such as newly diagnosed breast cancer patients, young unaffected high-risk women, and males are also reviewed. Alternative forms of and adjuncts to traditional face-to face genetic counseling are discussed. PMID- 17917142 TI - Psychosocial and behavioral impact of genetic counseling and testing. AB - Over a decade has passed since the clinical availability of BRCA1/2 mutation testing for Hereditary Breast and Ovarian Cancer (HBOC). The purpose of this article is to review key areas of psychosocial and behavioral research related to genetic counseling and testing for BRCA1/2 mutations. Special attention will be given to understudied issues within each of these key areas. Where appropriate, the article will also highlight the clinical and research experiences of the authors. The first area that will be reviewed is the impact of genetic testing on psychological well-being. This will be followed by a brief discussion of a practical assessment strategy for psychosocial distress in clinical settings. Next, published data on the uptake of risk management options based on genetic testing results as well as the psychosocial impact of these behaviors will be reviewed. Thirdly, research focused on understanding the decision making at various points in the genetic counseling and testing process will also be examined. Finally, the available research on genetic counseling and testing in minority communities will be presented. By recognizing and addressing the psychosocial and behavioral issues faced by patients undergoing BRCA1/2 genetic counseling and testing, researchers and providers have the potential to maximize opportunities for prevention, early detection, and healthy coping. PMID- 17917143 TI - International perspectives on genetic counseling and testing for breast cancer risk. AB - Familial cancer services have been developed in many countries in response to a rapidly evolving demand for genetic counseling and testing for breast cancer risk. This article presents a synthesis of the literature on international aspects of genetic counseling and testing and discusses similarities and differences in the provision of genetic counseling and testing, taking seven countries with well established familial cancer services as a case study (Australia, Canada, France, Germany, Netherlands, UK and US). Potential international differences are discussed in terms of: provider and patient attitudes to genetic counseling and genetic testing; utilization rates of genetic testing and prophylactic surgery; and the psychological impact of genetic testing for breast cancer risk. The comparative analyses of utilization rates and the psychological impact of testing indicate a wide range of variability in uptake rates and psychological outcomes, most likely reflecting sample variability and methodological differences in measurement. International comparison studies using controlled designs would be required to ascertain whether international differences exist, and to disentangle the differential role of clinical, individual and family context factors, on the one hand, and the cultural and health system-related factors unique to particular countries, on the other. PMID- 17917144 TI - Life after BRCA1/2 testing: family communication and support issues. AB - The process of genetic testing is often deemed a family affair. Several studies have indicated that individuals undergo BRCA1/2 testing not only to learn about their own cancer risks and options for screening and prevention, but also to gather information for potentially at-risk relatives. However, many individuals are not prepared for the medical and emotional implications that accompany the genetic testing process. This can be complicated by a moral or ethical obligation to disclose result status to other family members. Several characteristics including gender, BRCA1/2 carrier status, and cultural and ethnic background may influence the communication process between the proband and his/her potentially at-risk kin. In addition, the age of family members and their degree of relatedness may affect whether or not they are told the results of their relative's genetic testing. While genetic providers have an obligation to inform individuals of the implications of BRCA1/2 test results for at-risk relatives, they must also strive to respect and maintain autonomy and confidentiality. This paper will examine the characteristics that influence the disclosure of BRCA1/2 test results to relatives. In addition, methods of post-test support and follow up to facilitate the disclosure process for patients and their family members as well as foster positive communication, will be discussed. PMID- 17917145 TI - A principle-based approach to ethical issues in predictive genetic testing for breast cancer. AB - Genetic breast cancer susceptibility testing presents ethical challenges for healthcare providers and their patients. The familial aspects of genetic information, recognition of DNA as a shared history and present common thread for all people and widespread misunderstandings of genetic tests all contribute to these challenges. In this article an ethical framework internationally developed as a charter for medical professionalism is used to guide approaches to ethical dilemmas of breast cancer genetic testing. Specifically, three ethical principles are explored as they relate to testing: primacy of patient, patient autonomy, and social justice. Approaching breast cancer genetic testing from this framework could help to ensure thoughtful and ethical practices in this rapidly evolving field. PMID- 17917147 TI - Mammary stem and progenitor cell regulation. AB - Although mammary stem cells were first identified almost two decades ago, recent progress in the characterization of mouse and human mammary stem and progenitor cells has provided new insight into the regulation of their developmental hierarchy. In this review, we discuss the parameters that distinguish stem and progenitor cells from differentiated cells, as well as the signal transduction pathways that drive multilineage expansion. Lastly, we present the utility of modulating stem cell self-renewal through the use of inhibitors of the Notch and Wnt pathways. PMID- 17917148 TI - Stem cells in chronic myeloid leukaemia. PMID- 17917149 TI - Regulation of the self-renewal ability of tissue stem cells by tumor-related genes. AB - Stem cells are defined by both their ability to produce stem cells, a property known as self-renewal, and their ability to give rise to differentiated progeny. One of the hallmarks of tissue stem cell is its adoption of a non-dividing, undifferentiated state called quiescence. The ability to sustain quiescence is crucial for stem cell function in several tissues. In this review, we discuss how tissue stem cell properties are affected by the products of tumor-related genes such as ATM, PTEN and FOXO. Recent advances in stem cell research achieved using mouse genetics have provided novel evidence that numerous tumor-related genes, which are known to control genomic stability, cell proliferation and survival, are also closely associated with the regulation of tissue stem cell self-renewal. These findings support the notion that tissue stem cells and cancer cells share common properties. Further investigation of stem cell regulation by tumor-related genes may pave the way for successful stem cell-based approaches to regenerative medicine and cancer therapy. PMID- 17917150 TI - Stem cells of the melanocyte lineage. AB - Melanocytes are pigment-producing cells responsible for coloration of skin and hair. Studies using mouse models have allowed identification of putative melanocyte stem cells within the hair follicle and understanding of hair graying caused by abnormal melanocyte stem cell maintenance. The malignant transformation of melanocytes results in melanoma, the sixth most common cancer in the United States. Recent studies have offered compelling evidence for the existence of cancer stem cells in numerous tumor types, including melanoma. In this review we provide an overview of some of the current findings on follicular melanocyte stem cells, the genetic pathways involved in their regulation and maintenance, and discuss recent studies that support the existence of cancer stem cells in melanoma. PMID- 17917151 TI - Markers in normal and cancer stem cells. AB - In an effort to better understand and address the challenges of cancer research and treatment, a new model of tumorigenesis is being developed - the cancer stem cell model. Building upon traditional concepts of cancer and stem cells, this model is intended to shed new light on the continued struggle with treatment issues such as tumor drug-resistance and recurrence. This review describes the cancer stem cell model with an emphasis on markers that represent the "stemness" phenotype. A thorough understanding of normal and cancer stem cells is necessary for a precise delineation of cancer stem cells. The objective of such an improved delineation is to develop targeted therapy for selective elimination of cancer stem cells with minimal toxicity to normal stem cells. Specific targeting of cancer stem cells has proved to be a significant challenge due to the commonality of many markers between normal and cancer stem cells. However, research in the area of cancer biomarkers is slowly, but steadily, progressing. PMID- 17917152 TI - Unraveling the complex nature of prostate cancer stem cells. AB - The identification of distinct prostate cancer stem cell biomarkers is necessary as researchers attempt to isolate, characterize, and therapeutically target these tumor initiating cells. However, in reading the current literature it is frequently difficult to discern which biomarkers and cellular characteristics identify prostate cancer stem cells as opposed to the general population of prostate cancer cells or normal prostate stem cells. Within this review, we address this issue by dissecting out the cell surface markers, adhesion and cytoskeletal proteins, signaling pathways, epigenetic modifications, and the effects of androgens that are specific to and help to define the prostate cancer stem cell. PMID- 17917153 TI - Stem cell-like cancer cells in cancer cell lines. AB - Both stem cells and cancer cells are thought to be capable of unlimited proliferation. Moreover, a small number of cancer cells express stem cell markers, including CD133 and ATP-binding cassette transporters, by which the cells can pump out specific fluorescence dyes, such as Hoechst33342, as well as anti-cancer drugs, suggesting that either cancer cells resemble stem cells or cancers contain stem cell-like cancer cells, called "cancer stem cells (CSCs)". Using the common characteristics of tissue-specific stem cells, it was demonstrated that many types of tumors and cancer cell lines contain CSCs, which self-renew, express stem cell markers, and are tumorigenic. It was also shown that CSCs are resistant to anti-cancer drugs and irradiation. Thus CSCs might be a crucial target for the therapy. Because tumors contain CSCs and recruited normal stem cells, both of which contribute to tumorigenesis, it is difficult to separate CSCs from tumors. By contrast, cancer cell lines do not have any contaminating normal stem cells that quickly loose mulitpotentiality and differentiate in normal culture condition, suggesting that cancer cell lines could be an attractive alternative source of cells for CSC research. In this review I summarize the recent progress in CSC research using cancer cell lines. PMID- 17917154 TI - Hepatocellular stem cells. AB - The liver has enormous regenerative capacity. Restitution of the liver in response to different injuries involves proliferation of cells at different levels of liver lineage. Mature hepatocytes, which are normally dormant, could undergo rapid replication with a near infinite capacity to proliferate. When the replication of mature hepatocytes is inhibited, a reserve compartment of bipotential hepatic progenitor/stem cells is activated. The degree of activation appears to correlate with the degree of inflammation and stage of chronic liver disease. Deregulation of key regulatory signaling pathways such as transforming growth factor-beta, Wnt, hepatocyte growth factor, insulin-like growth factor, transforming growth factor-alpha and epidermal growth factor in this progenitor/stem cell population could give rise to HCC. Further understanding of these key signaling pathways and the molecular and genetic alterations associated with HCC could provide major advances in new therapeutic and diagnostic modalities. PMID- 17917155 TI - Biomarkers of WNT signaling. AB - WNT signaling plays a key role in the developing embryo and in the maintenance of a stem/progenitor cell compartment in adult tissues. However, WNT signaling is also believed to play an important role in carcinogenesis. WNT signaling may be activated directly through WNT ligand overexpression, or more often through mutations downstream in the WNT signaling cascade. The common endpoint is an inappropriate expansion of a stem cell compartment and proliferation of more differentiated daughter cells which ultimately acquire additional mutations leading to the development of a malignant, invasive cancer cell. It is likely that WNT signaling is at the core of many human cancers, but no definitive biomarker of WNT activity has been established. Furthermore, while therapies targeting WNT-responsive gene products such as COX-2 and VEGF have been developed, no therapy directly targeting WNT signaling itself has yet made it into the clinic. We present here a brief summary of the WNT signaling cascade, and focus particularly on markers of WNT activity that have diagnostic, prognostic, or therapeutic relevance. Finally, we propose the development of novel ligands of the steroid hormone family of nuclear receptors to be used as therapies that specifically target and inhibit WNT/beta-catenin/TCF-mediated transcriptional activity. PMID- 17917156 TI - Longitudinal brain corticotropin releasing factor and somatostatin in a transgenic mouse (TG2576) model of Alzheimer's disease. AB - Neuropeptides corticotropin releasing factor (CRF) and somatostatin (SRIF) are substantially decreased in cortical regions of Alzheimer's disease (AD) post mortem brain tissue. The accumulation of amyloid-beta (Abeta) in AD brain has been postulated to be neurotoxic. Using male Tg2576 mice transgenic over expressing amyloid-beta protein precursor (APP), we examined brain concentrations of CRF and SRIF at 12, 18 and 24 months. Mice were evaluated for locomotor activity and spatial memory. The APP mice had continued increased locomotor activity from 6 months of age compared to controls. Spatial memory was impaired beginning at 12 months in the APP mice relative to controls. APP mice at 24 months had a significantly higher number of amyloid plaques when compared to the 12 and 18 month time points. Brain concentrations of SRIF and CRF were significantly altered in a number of cortical and sub-cortical brain regions relative to controls, but in most regions were increased rather than decreased as in clinical AD. This data shows that although the insertion of the APP gene does cause age dependent increase in plaque load, it does not cause a change in regional neuropeptides consistent with AD, suggesting that neuropeptide changes in AD are not solely due to Abeta load. PMID- 17917157 TI - Androgens in the etiology of Alzheimer's disease in aging men and possible therapeutic interventions. AB - Animal experiments and cell biology studies have provided evidence that both estrogens and androgens can play a protective role against Alzheimer's disease (AD) related neurodegeneration. Males who become hypogonadal in later life often report problems with their memory. Lower than normal testosterone levels have also been detected in patients prior to the onset of AD, as well as in younger late-onset male AD patients, when compared to appropriate controls. The results of some small clinical trials suggest that testosterone can improve cognitive function in andropause. Although such improvement in cognitive function is subtle, patients on testosterone replacement therapy have reported memory improvements in both declarative and procedural domains. In contrast, there is no clinical evidence to date which suggest that the hormone dihydroepiandrosterone (DHEA) can improve cognitive function. Rises in the levels of the gonadotropins, follicle stimulating hormone (FSH) and luteinizing hormone (LH), have been associated with AD, but the clinical effects of reducing their levels remain to be determined. We hypothesize that androgens, gonadotropin modulators, or perhaps selective androgen receptor modulators may be useful components of therapy aimed at preventing the onset or delaying the progression of AD in male patients. PMID- 17917158 TI - Thiophilic interaction chromatography (TIC) of amyloid-beta protein precursor. AB - Neuritic plaques, one of the diagnostic characteristics of an AD, contain extracellular deposits of amyloid-beta (Abeta) derived from amyloid-beta protein precursor (AbetaPP). The objective of this study was to extract AbetaPP out of HEK293 cells and to purify it. Two procedures were chosen for purification of AbetaPP: Thiophilic Interaction Chromatography (TIC) and molecular sieving. Using Superdex 75, Superose 12, and Fractogel gel matrices, AbetaPP was isolated on HPLC. The chromatograms illustrate the purification of AbetaPP. Our method describes a new and elegant way for the extraction and purification of AbetaPP from HEK293 cell lines using thiophilic interaction chromatography (TIC). PMID- 17917159 TI - Elevated serum C-reactive protein concentration in Bosnian patients with probable Alzheimer's disease. AB - Studies indicate that inflammatory mechanisms may play an important role in the pathogenesis of Alzheimer's disease (AD). C-reactive protein (CRP), marker and mediator of inflammation, has been detected in lesions typical for the affected areas of AD brain. There have been conflicting reports on serum CRP concentration in AD. Scarce data exist on association of CRP and measures of adiposity in AD patients. Thus, we investigated serum CRP concentration in fifteen overweight institutionalized patients with probable AD and fifteen age-matched control subjects. Body mass index (BMI) and waist/hip ratio (WHR) were calculated for each subject included in the study. Age, systolic and diastolic blood pressure, BMI and WHR did not differ significantly between the two groups. Serum CRP concentration was significantly higher in patients with AD compared to controls (p<0.0001). Although not significant, positive correlations between serum levels of CRP and BMI and WHR were found. Obtained results support the notion that low grade inflammation is present in patients with AD. Absence of significant association between CRP and measures of total and central adiposity in overweight AD patients needs further investigation and explanation. PMID- 17917160 TI - About the presence of hemosiderin in the hippocampus of Alzheimer patients. PMID- 17917161 TI - Cajal's contributions to the study of Alzheimer's disease. AB - Last year 2006, we commemorated two important events in the history of Neuroscience. One hundred years ago, on November 3, Alois Alzheimer (1864-1915) presented the first case of a patient with symptoms of a disease that later would be called Alzheimer's disease. One month later, on December 10, Santiago Ramon y Cajal (1852-1934) and Camilo Golgi (1843-1926) received the Nobel Prize "in recognition of their work on the structure of the Nervous System". These facts seem not to be related, but working in the Museum Cajal we found 37 histological preparations of material from patients suffering from Alzheimer's disease, revealing that Cajal also studied this disease. This paper deals with Cajal's contribution to the study of Alzheimer's disease and it is fully illustrated by original pictures of Cajal's slides preserved in the Cajal Museum, Madrid. PMID- 17917164 TI - Lipoic acid and N-acetyl cysteine decrease mitochondrial-related oxidative stress in Alzheimer disease patient fibroblasts. AB - In this study, we evaluated the effect of lipoic acid (LA) and N-acetyl cysteine (NAC) on oxidative [4-hydroxy-2-nonenal, N(epsilon)-(carboxymethyl)lysine and heme oxygenase-1] and apoptotic (caspase 9 and Bax) markers in fibroblasts from patients with Alzheimer disease (AD) and age-matched and young controls. AD fibroblasts showed the highest levels of oxidative stress, and the antioxidants, lipoic acid (1 mM) and/or N-acetyl cysteine (100 microM) exerted a protective effect as evidenced by decreases in oxidative stress and apoptotic markers. Furthermore, we observed that the protective effect of LA and NAC was more pronounced when both agents were present simultaneously. AD-type changes could be generated in control fibroblasts using N-methylprotoporphyrin to inhibit cytochrome oxidase assembly indicating that the the oxidative damage observed was associated with mitochondrial dysfunction. The effects of N-methylprotoporphyrine were reversed or attenuated by both lipoic acid and N-acetyl cysteine. These data suggest mitochondria are important in oxidative damage that occurs in AD. As such, antioxidant therapies based on lipoic acid and N-acetyl cysteine supplementation may be promising. PMID- 17917163 TI - Mitochondrial effects of lipid-derived neurotoxins. AB - Mitochondria play a pivotal role in the life and death of cells and likely contribute importantly to the initiation or progression of many neurodegenerative disorders. Brain is especially vulnerable to lipid peroxidation because of its enrichment in polyunsaturated fatty acids relative to other organs. Multiple studies demonstrate the elevation of numerous lipid peroxidation products and ensuing lipid-derived carbonyls in Alzheimer's disease. In this review, we examine the data describing the effects of lipid-derived carbonyls such as 4 hydroxy-trans-2-nonenal (HNE) and acrolein upon the function of brain mitochondria as well as review the detoxification routes of these carbonyls. From these data, we show that the physical-chemical properties of these carbonyls influence their effects on mitochondria and that multiple essential mitochondrial functions are altered by these endogenous reactive carbonyls. Lastly, we examine the role of mitochondrial aldehyde detoxification pathways and their potential role in the development of AD. PMID- 17917162 TI - Amyloid-beta-induced mitochondrial dysfunction. AB - As an important molecule in the pathogenesis of Alzheimer's disease (AD), amyloid beta (Abeta) interferes with multiple aspects of mitochondrial function, including energy metabolism failure, production of reactive oxygen species (ROS) and permeability transition pore formation. Recent studies have demonstrated that Abeta progressively accumulates within mitochondrial matrix, providing a direct link to mitochondrial toxicity. Abeta-binding alcohol dehydrogenase (ABAD) is localized to the mitochondrial matrix and binds to mitochondrial Abeta. Interaction of ABAD with Abeta exaggerates Abeta-mediated mitochondrial and neuronal perturbation, leading to impaired synaptic function, and dysfunctional spatial learning/memory. Thus, blockade of ABAD/Abeta interaction may be a potential therapeutic strategy for AD. PMID- 17917165 TI - Concussion and sports. PMID- 17917166 TI - The effects of explanatory style on concussion outcomes in sport. AB - Individuals with an optimistic explanatory style have generally been linked with improved mental and physical health across a variety of chronic and serious conditions. The purpose of the current study was to examine the effects of explanatory style on recovery time and number of sport-related concussions suffered in the last 12 months. University varsity athletes (n=170) suffering from at least one concussion over the last 12 months from six contact or collision team sports completed both the Attributional Style Questionnaire and the Sport History Questionnaire. The results indicated that athletes with an optimistic explanatory style took longer to recover than athletes with a pessimistic or average explanatory style. More specifically, optimistic athletes who suffered a complex concussion (requiring more than 7 days to recover) took significantly longer to return to play. However, the results showed that explanatory style did not influence whether an athlete suffered subsequent concussions. Overall, the current results can be used to better understand the psychology of concussions, as well as concussion prevention efforts and management strategies. PMID- 17917167 TI - Use of computer based testing of youth hockey players with concussions. AB - Concussion is a potentially serious injury for athletes. Recent statistics suggest that approximately 300,000 sports-related traumatic brain injuries occur annually in the United States. Soccer, rugby, football, and ice hockey are all considered high-risk team sports for concussion. Hockey-related concussions are of particular concern in Canada, where over 500,000 players compete annually in ice hockey. The United States is now registering similar numbers of players. Return to play issues are one of the most difficult issues for physicians caring for concussed athletes. The advent of computerized neuropsychological testing adds another tool to assist in this process. It also appears to enhance the education process for players, coaches, and parents on the potential seriousness of concussion for these young athletes. PMID- 17917168 TI - Concussion sideline management intervention for rugby union leads to reduced concussion claims. AB - The effectiveness of a concussion management education programme (CMEP) in rugby in reducing the number and cost of concussion/brain injury (CBI) moderate to serious claims (MSC) was assessed. A RugbySmart educational video and a sideline concussion check (SCC) tool comprised the CMEP. Over 30,000 SCC, providing information on management of suspected concussion among community level rugby players prior to seeking medical treatment, were distributed from July 2003 to June 2005. Each year approximately 10,000 coaches and 2,000 referees participated in RugbySmart. From 2003 to 2005 new rugby CBI MSC reduced by 10.7% (actual) and 58.2% (forecast). Rugby player numbers, new non-sport CBI MSC and new sport MSC all increased by 13.6%, 16.9% and 24.6% respectively in the same period. The median number of days between CBI injury and the player seeking medical treatment decreased from six days to four days. Cost savings after CMEP were 690 USD dollars, 690 (actual) to 3,354,780 USD dollars (forecast). The two-year cost of CMEP was 54,810 USD dollars returning 12.60 USD dollars (actual) and 61.21 USD dollars (forecast) for every 1 USD dollar invested (ROI). CMEP provided community coaches and managers with education on minimum best practice for managing suspected concussion, contributed towards ROI and savings for CBI MSC in rugby. PMID- 17917169 TI - Use of the ICECI and ICD-10 E-Coding structures to evaluate causes of head injury and concussion from sport and recreation participation in a school population. AB - The purpose of this study was to identify the most common causes of head injury and associated symptoms of concussion in a population of school children (n=1,372,979). Using standardized injury report forms, we identified the mechanisms of head injury associated with various sports/recreation activities and assessed each injury for the presence of concussion symptoms. Head injury reports (n=7,765) were coded using the WHO's ICECI and ICD-10 E-codes. 1,338 Cases reported symptoms of concussion. The majority of head injuries occurred during School Free-Play/Recess (59.8%). Combative Sports and Wheeled Non-Motored Sports were the activities most often associated with concussion symptoms (rate of concussion (RC): 48.3% and 44.4%, respectively, p<0.001). School Free Play/Recess and Physical Education Classes were significantly less likely to have head injuries associated with concussion (RC: 16.0%, p<0.001, and 12.4%, p=0.034, respectively). The most common causes of head injury were (1) Struck by an Object (24.9%) and (2) Falling on the Same Level (22.8%). Falling from a Transport was the only etiological code significantly associated with concussion symptoms after head injury (RC: 28.7%, p<.001). Results were similar when using the two coding structures in combination. Prevention efforts should focus on activities where children are moving at high speeds since these are more likely to cause a concussion when a head injury occurs. PMID- 17917170 TI - Regulatory and autoregulatory physiological dysfunction as a primary characteristic of post concussion syndrome: implications for treatment. AB - Although most patients with concussion recover within days to weeks, a small but significant minority develop persistent signs and symptoms of post-concussion syndrome (PCS). The standard treatments of PCS, rest and cognitive adaptation, have limited effectiveness. PCS patients are advised not to exercise because of the concern for symptom exacerbation. Prolonged rest, however, leads to deconditioning (especially in athletes) and may cause secondary effects including depressive symptoms. Concussion is associated with metabolic and physiological changes in the brain and in other organ systems (for example, autonomic function of the heart and altered cerebral autoregulation, sleep, and circadian rhythms). We propose that PCS results from ongoing central and systemic physiologic regulatory dysfunction after traumatic brain injury (TBI) and we further propose that this physiologic dysfunction may be reduced or alleviated by individualized controlled sub-symptom threshold aerobic exercise rehabilitation. PMID- 17917171 TI - The relation between post concussion symptoms and neurocognitive performance in concussed athletes. AB - The objective of this study was to examine differences in neurocognitive performance between symptomatic concussed athletes, a group of concussed athletes with no subjective symptoms, and a non-concussed control group of athletes. All concussed athletes were evaluated within one week of injury using the ImPACT computerized test battery. Results indicate that concussed athletes who denied subjective symptoms demonstrated poorer performance than control subjects on all four composite scores of the ImPACT test batters (Verbal Memory, Visual Memory, Reaction Time and Processing Speed However, the concussed but asymptomatic group demonstrated significantly better performance than did the concussed and symptomatic group. Thus, concussed athletes who did not report subjective symptoms were not fully recovered based on neurocognitive testing. This study underscores the importance of neurocognitive testing in the assessment of concussion sequelae and recovery. PMID- 17917172 TI - Contributions of functional magnetic resonance imaging (fMRI) to sport concussion evaluation. AB - Mild traumatic brain injury (mTBI) in contact sport is a problem of such magnitude that improved approaches to diagnosis, investigation and management are urgent. Concussion has traditionally been described as a transient, fully reversible, cerebral dysfunction. However, this seemingly 'mild' injury sometimes results in long-lasting and disabling post-concussion symptoms (PCS) and abnormal neuropsychological profiles characteristic of frontal and/or temporal lobe dysfunction. At present, the pathological changes following concussion remain unclear, but it is now widely accepted that concussion results mainly in functional disturbance rather than structural damage. Therefore, functional imaging techniques can help in demonstrating brain abnormalities undetectable by structural imaging methods. This paper will review the use of functional magnetic resonance imaging (fMRI) in studies of concussion. Our existing and ongoing fMRI studies will be described as examples to highlight the potential and contribution of this non-invasive functional neuroimaging technique in the assessment of sports-related concussion and its management. PMID- 17917173 TI - Post concussion syndrome ebb and flow: longitudinal effects and management. AB - This research identified persistent post concussion symptoms (PCS) in a group of 20 adult subjects. PCS generally lasted for two years with a mean of 3.35 years. Typical symptoms included physical and cognitive fatigue, depressive behaviors, sensitivity to noise, social withdrawal, irritability, concentration and problem solving difficulties, loss of libido and much difficulty making decisions at even the simplest strategic level. They represented a hard core group for whom the original symptoms persisted well beyond the 6~month period. Participants identified their PCS according to sensory, somatic affective and cognitive items immediately following their trauma (01) and two years later (02). Counseling and psychotherapy intervention took place between 01 and 02. Items on the PCS schedules and the Beck Depression Inventory (II) demonstrated significant decline in the presence of overall symptoms most noticeably in reduction of agitation, irritability and suicidal wishes. However, subjects throughout generally experienced the feeling that they were being punished which equated with behaviors comparable with learned helplessness. The PCS group considered themselves to be different people after trauma. They had different goals, changing lifestyle, relationships and employment and were more often in a dependent state. Comparability with other conditions such as PTSD and chronic fatigue syndrome (CFS) was demonstrated by individuals who experienced persistent and invasive post concussion symptoms. PMID- 17917174 TI - Retest reliability in adolescents of a computerized neuropsychological battery used to assess recovery from concussion. AB - We examined in a group of 15-year-old adolescents the retest reliability over one week of 7 subscales of the Automated Neuropsychological Metrics (ANAM), a computerized battery based on standard neuropsychological test measures that is one of several such batteries available to assess concussion effects. Since the principle behind these computerized batteries is to assess athletes before injury and after injury to determine the level of deficit and whether the individual is safe to return to play, it is critical that such batteries have excellent retest reliability. Retest reliability of the ANAM was good, especially for the aggregate of throughput scores, reaching 0.87, but lower for individual subtests, especially for those measuring only speed of processing. Thus, the ANAM aggregated score appears to have robust reliability for cognitive measures involving memory and attention in 15-year-olds. Limitations related to assessing return-to-baseline after concussion in adolescents are discussed. PMID- 17917175 TI - Sociotechnologies of care: visions and realities. PMID- 17917176 TI - Designing and evaluating healthcare ICT innovation: a cognitive engineering view. AB - Many healthcare ICT innovations fail in practice because of a failure to take into account user needs. User needs should be identified broadly, encompassing different levels of organization of the healthcare system and different stakeholder concerns. Full-scale simulators are starting to be used to help in the design and evaluation of novel biomedical devices and displays. Although promising, simulators have significant technical and operational limitations for this purpose and they do not address important aspects of the sociotechnical systems context in which healthcare ICT will be embedded. This argument is illustrated with a case study in which advanced auditory displays for patient monitoring were successfully evaluated in a full-scale patient simulator, but many further questions remain prior to successful translation to practice. PMID- 17917177 TI - Does it work on Sundays, too? Healthcare technology for older people. AB - The development of assisted living technology today lies within the realm of ambient computing, making assistance automatic and the systems invisible. Unfortunately, this invisibility is also the reason why the users of these systems have no means to remedy even very simple fault situations. By focusing on the needed complementarities between user control and automation, we identify three main issues which are critical when introducing new technology in the homes of older people: Individual need for representation of data, the need for the user to construct a conceptual model of the system and the need for systems to change over time. PMID- 17917178 TI - Embracing standard treatment manuals: information transfer to primary health workers in Papua New Guinea. AB - Papua New Guinean primary health care workers describe their Standard Treatment Manuals as like "a teacher" or "a doctor". This paper explores this glowing reference, looking at how Standard Treatment Manuals are utilised, through data collected in a study which identifies what influences primary health care workers in Papua New Guinea to access and utilise information for diagnostic and treatment decisions. In addition to presenting a unique perspective on this method for transferring information to health workers across the world, this paper presents an opportunity to consider some of the factors which both enable and inhibit the process of providing information to health workers in a non western culture. PMID- 17917179 TI - Complexity and its implications for health systems implementation. AB - The National Programme for IT in England is an ambitious programme comprising a variety of systems from the tried and tested to the new based on new government policy that is being implemented at the same time. For such a large and complex programme, it is not surprising that there are a variety of outcomes emerging. While there are many successes, there are also delays and concerns. This paper looks at two very different systems in the programme (Choose and Book and PACS). It compares and contrasts their implementation within a health community and identifies implications, based on complexity theory, this has for the choice of approach to implementation of associated change. PMID- 17917180 TI - Structuration and sensemaking: frameworks for understanding the management of health information systems in the ICU. AB - This paper will describe two alternate conceptual frameworks (i.e. Structuration and Sensemaking) that will help to describe and provide insight into how best to implement health information systems in ICUs throughout the globe. Structuration and sensemaking are two competing ways to view the social world within hospitals. To examine the impact of information technology in health care organizations, it is important to explore the dynamic interplay between clinical decisionmaking, outcomes of HIT implementation, and individual characteristics of the organizational setting. The adaptation of information technology within health care organizations is by its very nature quite complex. The recursive pattern of social interactions that shape the implementation of technologies within that setting is key. Structuration theory provides an understanding of human work as social interaction within that organizational culture, mediated by artifacts such as tools, language, rules and procedures, and open to change. The ICU provides multiple opportunities for sensemaking. It involves caring for multiple patients simultaneously; is subject to high levels of uncertainty and is provided under significant time constraints. It is highly interdependent work, necessitating shared sensemaking as well as individual sensemaking. Sensemaking is made partially visible in this context as clinicians communicate to each other what they think is the cause of the patient's symptoms and how to treat them in the form of discussions about patient care, consultation requests, ancillary testing, and the electronic medical record. The collaborative nature of work in the ICU lends itself to the application of sensemaking and structuration theories. PMID- 17917181 TI - Whose work practice? Situating an electronic triage system within a complex system. AB - An electronic triaging system was introduced into a busy children's hospital emergency department. Within 18 months of its introduction, amidst complaints from staff about patient safety related to work slow downs, a decision was made to stop using the system. In this paper we examine issues that arose with the introduction of the electronic triaging system, and discuss these in relation to decision making in complex systems. We suggest that difficulties with the triage system resulted partly because data resulting from the triage encounter are used in several domains including the care domain, the access domain, the federal equity and accountability domain, the local accounting and quality domain and the research domain, each of which has different primary stakeholders, with varying needs. Greater attention to identification of data requirements for each of these domains and acknowledgement of varied stakeholder interests prior to software selection and implementation may improve future implementations. PMID- 17917182 TI - Evaluating the implementation and use of a computerized physician order entry system: a case study. AB - A study at the urology clinic, Frederiksberg Hospital Denmark, the clinicians working procedures, when prescribing, dispensing and administrating drugs using a computerized physician order entry system (CPOE) showed an overwhelming majority of the clinicians who believed it does support the patient safety and quality of treatment despite the CPOE was not used to the expected degre. PMID- 17917183 TI - Telemedical teamwork between home and hospital: a synergetic triangle emerges. AB - The aim of this paper is to describe the effect on the clinical cooperation when introducing video consultations in the home of the patient. The study was conducted as a Participatory Design process containing workshops, field studies, clinical experiments and pilot tests involving participants from the Danish primary and secondary care sectors as well as patients and relatives. The video consultation set-up constitutes a new organisational way of working, described as "a new triangle", based on immediate inter-individual cooperation and team-work. In the triangle, competences were combined which led to a more holistic treatment and a more active role of the patient. Furthermore, a spreading of knowledge between all participants was seen, resulting in an upgrading of the competences of especially the visiting nurse. The introduction of a real-time, on-line link between hospital and home constitutes the basis for simultaneous communication between all participants, resulting in a "witnessing" situation potentially securing or even enhancing quality of treatment. PMID- 17917184 TI - ICT & OTs: a model of information and communications technology acceptance and utilisation by occupational therapists (part 2). AB - The research reported in this paper describes the development, empirical validation and analysis of a model of technology acceptance by Australian occupational therapists. The study described involved the collection of quantitative data through a national survey. The theoretical significance of this work is that it uses a thoroughly constructed research model, with one of the largest sample sizes ever tested (n=1605), to extend technology acceptance research into the health sector. Results provide strong support for the model. This work reveals the complexity of the constructs and relationships that influence technology acceptance and highlights the need to include sociotechnical and system issues in studies of technology acceptance in healthcare to improve information system implementation success in this arena. The results of this study have practical and theoretical implications for health informaticians and researchers in the field of health informatics and information systems, tertiary educators, Commonwealth and State Governments and the allied health professions. PMID- 17917186 TI - Analysis of communicative behaviour: profiling roles and activities. AB - In this paper, we discuss how profiles of communicative behaviour can be used to present and analyse information about role activity recorded through structured observation of specific situations. The role activities are encoded as distinctive speech acts. Example profiles resulting from the analysis of three clinicians' communicative behaviour during pre-rounds meetings and ward rounds are given. The examples are based on an observational study performed at a Norwegian university hospital. One fifth-year medical student spent 20 days in two different hospital wards, following 7 physicians from one to seven days each. The observer recorded data from several ward situations such as pre-rounds meetings, ward rounds, and discharge situations. The data was recorded by means of an observation form consisting of a mixture of codes and free-text fields. The data has been post-processed by associating each event with one communicative act. The approach is an efficient and useful means for studying clinicians' information and communication patterns in hospital wards, which can serve as an important tool in the design of new clinical information systems. PMID- 17917185 TI - Initiators of interruption in workflow: the role of MDs and RNs. AB - The healthcare environment has been characterized as interrupt-driven with medical doctors (MDs) and registered nurses (RNs) receiving many interruptions during a shift. Previous research studies have focused on the recipient because of the negative impact on task performance. It is equally important to understand the initiator of an interruption to help design strategies to lessen the number of interruptions and the possible negatives consequences. The purpose of this instrumental study was to examine MDs and RNs as initiators of interruptions. Results of this study indicate that MDs and RNs initiate interruptions most often through face-to-face situations and use of the telephone. Strategies to successfully manage interruptions must consider both the role of initiator as well as the recipient in an interruption event. PMID- 17917187 TI - Pathology's front line - a comparison of the experiences of electronic ordering in the clinical chemistry and haematology departments. AB - Socio-technical approaches to health information systems evaluation are particularly relevant to the study of Computerised Provider Order Entry (CPOE) systems. Pathology services are made up of a number of departments each with unique and complex tasks and requirements. These different components of pathology have received very little research attention. This study used qualitative methods to identify key organisational and work process along with repercussions of the implementation of CPOE through a comparison of the Haematology and Clinical Chemistry departments of a hospital pathology service. The results focus attention on areas where the departments face similar challenges along with those areas where work practices diverged. This underlined the key importance of understanding the context and setting of pathology laboratories. The study also draws attention to the importance of cross departmental and multi-disciplinary negotiation in the implementation process and highlights the potential for technology to affect and be affected by the organisational context in which it is placed. PMID- 17917188 TI - Towards automated observational analysis of leadership in clinical networks. AB - Clinical networks are being increasingly employed to drive innovation in health services by encouraging multi-disciplinary clinical engagement in management processes. The effectiveness of a network, however, depends critically on the ability of its leader to coordinate group interactions. This paper discusses leadership of clinical networks, and in this context reviews technologies for analyzing the way team members interact in group conversations. This review will form the foundation for ongoing research to develop the profile of an effective clinical network leader, along with techniques and tools for evaluation and professional development. PMID- 17917189 TI - Constructing technology-in-use practices: EPR-adaptation in Canada and Norway. AB - Investigating two longitudinal ethnographic studies of managing gradual adaptation of electronic patient records in Canada and Norway, we conduct a cross case analysis of the enabling factors that support a continuous transformation of technology and health care practices. In line with previous research, our study shows that large-scale information systems adaptation in health care should be managed by a project-group including not only IT-developers, but also representatives of future users and management. While we also argue for the importance of these meetings, we complement and expand the notion of project meetings by providing a conceptualization of the essential aspects of these meetings. Our conceptualization is composed of continuous reflection-on-practice activities to construct technology-in-use practices. Reflection-on-action activities are internally initiated, and comprise critical reflections by the participants, who continuously evaluate and question work practices in relation to technology. PMID- 17917190 TI - Measuring nurses' time in medication related tasks prior to the implementation of an electronic medication management system. AB - A substantial proportion of nurses' work consists of medication related tasks undertaken in a current environment of high medication error rates. Electronic medication management systems (EMMS) are anticipated to address many of the factors which contribute to errors. These factors, for example illegible hand written drug orders, reputedly require nurses to spend additional time in clarification and discussion. We report an observational modified time and motion study of 44 registered nurses (215 hours of observation) in a major academic hospital to quantify the time nurses spend in medication related tasks prior to EMMS introduction. On average nurses spent seven minutes in an 8.75 hour shift clarifying or discussing medication issues. The majority of this time (5.4 mins) was spent talking with other nurses. Only 1.6 minutes was spent clarifying or discussing medication with doctors. Our results call into question some of the predicted efficiency benefits to nurses' work following EMMS introduction. PMID- 17917191 TI - Generation Y in healthcare: the need for new socio-technical consideration for future technology design in healthcare. AB - While identifying reasons for the failure of information communication technology (ICT) to transform the healthcare system and constructing models of better designed technology with socio-technical integration is relatively straightforward, implementing these solutions into the rapidly changing medical world has proven considerably more difficult. From a technologist's perspective, the promise of technology remains powerful. New technologies, with high level of socio-technical integration have long been considered as one of the most important factors to transform the medical world in order to deliver better and safer care. From the socio-cultural perspective, however, there is an equally powerful force, which has largely been ignored by the greater community: the entry of generation Y into the healthcare system. Generation Y has generated significant changes in many other industries. This powerful socio-cultural change within the healthcare system needs to be more clearly investigated to guide the design and implementation of sociotechnical integrated ICT solutions. This research-in -progress paper presents a methodological approach that both generates an in-depth understanding of generation Y and illuminates criteria that can be used to meaningfully identify the guiding principles for future socio technical integrated ICT design and implementation. It aims to make a significant contribution to the field of socio-technical approach to ICT design by alluding the audience to this new generation Y phenomenon in healthcare. It provides some preliminary data to support the need to consider generation Y in future ICT design in healthcare. PMID- 17917192 TI - A longitudinal study of usability in health care - does time heal? AB - We report from a longitudinal laboratory-based usability evaluation of a health care information system. A usability evaluation was conducted with novice users when an electronic patient record system was being deployed in a large hospital. After the nurses had used the system in their daily work for 15 months, we repeated the evaluation. Our aim was to inquire into the nature of usability problems experienced by novice and expert users, and to see to what extend usability problems of a health care information system may or may not disappear over time, as the nurses get more familiar with it - if time heals poor design. On the basis of our study, we present findings on the usability of the electronic patient system as experienced by the nurses at these two different points in time and discuss implications for evaluating usability in health care. PMID- 17917193 TI - Using blogging tools to help individuals record their experiences: an exploration and review of two commercial web applications in the Netherlands. AB - Blogs, short for "web logs," together with podcasts and wikis are currently important foci of general internet research. These three applications are part of the larger body of next-generation communication applications that comprises "Web 2.0." Within the specific area of health care, however, little attention has been devoted to understanding these technologies and how they are being used by lay health publics. In this article, I will discuss the emergent findings from a new project that looks at blogging interfaces as potential tools for disease prevention and health promotion. I use a literature review combined with "front stage" web analyses of two cases and interviews with the supporting institutions for these sites to discuss the relevant informatics questions that arise with respect to these applications. I further introduce the idea of "goal-oriented" blogging that is found in the first case study. Because this research project is still in preliminary phases, this should be viewed as an exploration into the topic and work in progress. In addition to raising questions, I will outline the important subsequent research steps. PMID- 17917194 TI - Heuristic evaluation performed by usability-educated clinicians: education and attitudes. AB - Heuristic evaluation is a usability testing method aiming to improve the user interface design. Traditionally, a panel of experts in usability and human factor issues evaluate and judge the compliance of computer software according to recognized usability principles, the heuristics. In this paper, we investigate clinicians' attitudes towards learning and performing a heuristic evaluation and present the procedure of educating the healthcare staff and their accomplishment of the evaluation. 18 clinicians were recruited for a 2-hours education and filled in a post-education questionnaire regarding their opinions of the evaluation method when applied by clinicians. Six of the clinicians participated later in a heuristic evaluation of a web-based virtual health record,Their time spent for evaluation and analysis of results was approximately four hours each. Opinions from the six "clinical evaluators" were gathered in an post-evaluation form and compared to the post-education questionnaire. The results of 18 clinicians indicate that there is an interest in learning and participating in such evaluations. Our interpretation is that it is feasible to educate healthcare staff to perform rapid usability inspections to locate usability defects and additionally emphasize the domain specific problems residing in health information systems. PMID- 17917195 TI - The medication advice-seeking network of staff in an Australian hospital renal ward. AB - Electronic medication systems may impact communication in hospital wards. To identify the ways in which communication patterns may be altered it is necessary to compare processes both before and after system introduction. This paper reports the use of a social network approach to examine the medication advice seeking network of an Australian hospital renal ward before the introduction of an electronic medication management system. A social network questionnaire was completed by 96% of staff members (doctors, nurses, allied health professionals and administrative staff) on the ward (n=45). Survey data were analysed to produce a sociogram to display the medication advice-seeking network of the staff in the ward. The results showed that there was a relatively low level of advice seeking about medication-related decisions and tasks. Most communication occurred within professional groups. Several key individuals were pivotal in providing advice both within and across professional groups. PMID- 17917196 TI - Using scenarios to capture work processes in shared home care. AB - Shared home care is increasingly common, and in order to develop ICT that support such complex cooperative work it is crucial obtain an understanding of the work routines, information demands, and other central preconditions at the clinical level before the development is initiated. Scenarios are proposed as a technique that can be useful for capturing work processes in shared home care and experiences from the Old@Home project are presented. The scenarios are useful not only in the initial phases of the development project but throughout the development process, improving the accessibility of end user requirements and usability issues for the design team, and as a basis for use cases and further design. PMID- 17917197 TI - Introducing an innovative, multidisciplinary concept of care - communication problems and possible solutions. AB - Introducing an innovative, multidisciplinary concept of care means the change of existing structures both in work and communication flows which often involves problems. In this paper we describe exemplary the communication problems which arose during the introduction of the multidisciplinary concept of Family-Centred, Individualized Developmental Care of Premature Infants and Newborns at the Children's Hospital Heidelberg. We suggest workflow adaptations to achieve timeliness of information and present ideas to prevent communication problems e.g. caused by inadequate knowledge of staff. We used interviews, analyzed video recordings and modified the Communication Observation Method [7] to analyze the present state of communication flows and structures. PMID- 17917198 TI - Enhancing immunization coverage through health information systems: a system dynamics approach. AB - This paper demonstrates how qualitative System Dynamics methodology can be used to provide a better understanding of health systems thus facilitating better development and design of computer-based health information systems. In an earlier paper by the same authors, system dynamics modeling and field study research methods are used to capture the complex and dynamic nature of the immunization process, to enhance the understanding of the immunization health care problems and to generate insights that may increase the immunization coverage effectiveness. Through qualitative modeling, causal loop diagrams which are used to show the key issues that need to be addressed when developing health information systems are drawn with the aim of improving the immunization services. The paper shows the benefits of using System Dynamics to understand systems with complex interactions thus facilitating the development of information systems that meet the stakeholder requirements. PMID- 17917199 TI - Differences in public and private sector adoption of telemedicine: Indian case study for sectoral adoption. AB - Telemedicine is the use of communication networks to exchange medical information for providing healthcare services and medical education from one site to another. The application of telemedicine is more promising in economically developing countries with agrarian societies. The American Telemedicine Association (ATA) identifies three healthcare services: clinical medical services, health and medical education, and consumer health information. However, it is not clear how these services can be adopted by different sectors: public and private. This paper looks at four Indian case studies, two each in public and private sectors to understand two research questions: Are there differences in telemedicine adoption between public and private hospitals. If there are differences: What are the differences in telemedicine adoption between public and private sectors? Authors have used the extant literature in telemedicine and healthcare to frame theoretical background, describe the research setting, present the case studies, and provide discussion and conclusions about their findings. Authors believe that as India continues to develop its telemedicine infrastructures, especially with continued government support through subsidies to private telemedicine initiatives, its upward trend in healthcare will continue. This is expected to put India on the path to increase its life expectancy rates, especially for it rural community which constitute over 70% of its populace. PMID- 17917200 TI - Conceptualisation of socio-technical integrated information technology solutions to improve incident reporting through Maslow's hierarchy of needs: a qualitative study of junior doctors. AB - Medical errors are common, especially within the acute healthcare delivery. The identification of systemic factors associated with adverse events and the construction of models to improve the safety of the healthcare system seems straightforward, this process has been proven to be much more difficult in the realism of medical practice due to the failure of the incident reporting system to capture the essential information, especially from the perspective of junior doctors. The failure of incidence reporting system has been related to the lack of socio-technical consideration for both system designs and system implementations. The main reason of non-reporting can be conceptualised through the motivation psychology model: Maslow's hierarchy of needs; in order to achieve a change in the socio-cultural domain for incident reporting. This paper presents a qualitative research methodology approach to generate contextual-rich insights into the socio-cultural and technological factors of incident reporting among junior doctors. The research illuminates the guiding principles for future socio technical integrated information communication technology designs and implementations. Using Maslow's hierarchy of needs as the conceptual framework, the guiding principles aim to design electronic incident reporting systems which will motivate junior doctors to participate in the process. This research paper aims to make a significant contribution to the fields of socio-technical systems and medical errors management. The design and implementation of the new incident reporting system has great potential to motivate junior doctors to change the culture of incident reporting and to work towards a safer future healthcare system. PMID- 17917201 TI - Socio-cultural issues and patient safety: a case study into the development of an electronic support tool for clinical handover. AB - This paper describes a case study into the development of an electronic support tool for clinical handover conducted in the Royal Hobart Hospital's Department of General Internal Medicine. By directly involving clinicians as co-participants in the development, and by conceptualising the system to be built as a support tool rather than as a 'total solution' this case study outlines the practical experience of dealing with a diversity of user requirements. The approach involved in-depth fieldwork to understand the factors and their inter relationships in clinical handover processes. From an analysis of the data generated key issues relating to work processes and potential impacts on patient safety were identified and discussed with clinicians. A support tool incorporating a series of design features aimed at improving patient safety and supporting existing work processes identified as important by the clinicians was developed. Through early and continual involvement of clinicians in the project, this case study highlights how socio-cultural analysis can be translated meaningfully (in terms of the end-users) into systems design. The paper aims to contribute to a stronger recognition within the domain of eHealth of user-centred approach to systems development for patient safety. PMID- 17917202 TI - Methodology for analysis of work practice with video observation. AB - Prior to developing and implementing health informatics systems it is essential to acquire information about the work practice. Interviews are often used but they don't always capture important details. In this project we used video cameras to observe the work practice of two medical secretaries at Department of Urology at Arhus University Hospital, Skejby. The project was carried out with a clear user influence in all phases of the project. The observation and the analysis focused on interruptions of the work processes, of which we found numerous that we categorized in three different types. The use of video and audio proved to be powerful tools for analysis of work practice, although it is very labor intensive method. PMID- 17917204 TI - Double complexes [Co(NH(3))(5)(H(2)O)](2)[Zr(3)F(18)].6H(2)O and [Co(NH(3))(6)](2)[Zr(3)F(18)].6H(2)O. AB - The structures of orthorhombic bis[pentaammineaquacobalt(III)] tetra-mu(2) fluorido-tetradecafluoridotrizirconium(IV) hexahydrate (space group Ibam), [Co(NH(3))(5)(H(2)O)](2)[Zr(3)F(18)].6H(2)O, (I), and bis[hexaamminecobalt(III)] tetra-mu(2)-fluorido-tetradecafluoridotrizirconium(IV) hexahydrate (space group Pnna), [Co(NH(3))(6)](2)[Zr(3)F(18)].6H(2)O, (II), consist of complex [Co(NH(3))(x)(H(2)O)(y)](3+) cations with either m [in (I)] or overline{1} and 2 [in (II)] symmetry, [Zr(3)F(18)](6-) anionic chains located on sites with 222 [in (I)] or 2 [in (II)] symmetry, and water molecules. PMID- 17917203 TI - Modelling the effect of limited or vulnerable resources on the use of computerised hospital information systems (CHISs) in South Africa. AB - A conceptual model of computerised hospital information system (CHIS) use has been developed, based on the results of case studies in four South African regional (level 2) hospitals, interviews with local experts, and related work on modelling and evaluation of health information systems. In addition to factors within hospitals, factors which reflect the decisionmaking and resource allocation processes at provincial level have also been included in the model. The applicability of the model is demonstrated through an analysis of the effects of limited or vulnerable resources on CHIS implementation and use at hospital level. Some potential approaches to overcoming these effects are suggested. PMID- 17917205 TI - A new pillared lithium bismuth phosphate, LiBi(7.37)P(3)O(19), with elliptical channels. AB - The structure of a new lithium bismuth phosphate, LiBi(7.37)P(3)O(19), consists of infinite OBi(4) edge-sharing tetrahedral chains in the ac plane, which form Bi(2)O(2) layers parallel to the b axis. They are sandwiched between PO(4) tetrahedral and Bi polyhedral layers. The PO(4)-Bi-PO(4) layers are bridged by columns formed by one Bi polyhedron flanked on each side by LiO(4) tetrahedra. This bridging Bi atom lies on a twofold axis, special position 4e of the C2/c space group. This arrangement creates pillared open elliptical channels parallel to [010]. PMID- 17917207 TI - Kolbeckite, ScPO(4).2H(2)O, isomorphous with metavariscite. AB - This study presents the first structural report of kolbeckite, with the ideal formula ScPO(4).2H(2)O (scandium phosphate dihydrate), based on single-crystal X ray diffraction data. Kolbeckite belongs to the metavariscite mineral group, in which each PO(4) tetrahedron shares four vertices with four ScO(4)(H(2)O)(2) octahedra and vice versa, forming a three-dimensional network of polyhedra. PMID- 17917206 TI - Isokite, CaMg(PO(4))F(0.8)(OH)(0.2), isomorphous with titanite. AB - This study presents the first structural report of natural isokite (calcium magnesium phosphate fluoride), with the formula CaMg(PO(4))F(0.8)(OH)(0.2) (i.e. some substitution of OH for F), based on single-crystal X-ray diffraction data. Isokite belongs to the C2/c titanite mineral group, in which Mg is on an inversion centre and the Ca, P and F/OH atoms are on twofold axes. The structure is composed of kinked chains of corner-sharing MgO(4)F(2) octahedra that are crosslinked by isolated PO(4) tetrahedra, forming a three-dimensional polyhedral network. The Ca(2+) cations occupy the interstitial sites coordinated by six O atoms and one F anion. PMID- 17917208 TI - Yttrium pyrogermanate, Y(2)Ge(2)O(7). AB - The structure of diyttrium digermanate, Y(2)Ge(2)O(7), has been determined in the tetragonal space group P4(3)2(1)2. It contains one Y, one Ge (both site symmetry 1 on general position 8b) and four O atoms [one on special position 4a (site symmetry ..2) and the remaining three on general positions 8b]. The basic units of the structure are isolated Ge(2)O(7) groups, sharing one common O atom and displaying a Ge-O-Ge angle of 134.9 (3) degrees , and infinite helical chains of pentagonal YO(7) dipyramids, parallel to the 4(3) screw axis. The crystal investigated here represents the left-handed form of the tetragonal R(2)Ge(2)O(7) compounds (R = Eu(3+), Tb(3+), Er(3+), Tm(3+) and Lu(3+)). PMID- 17917209 TI - SrZn(11): a new binary compound with the BaCd(11) structure. AB - Single crystals of strontium undecazinc, SrZn(11), were obtained when decomposing SrZn(2) under conditions of high pressure and high temperature. The new binary Sr Zn compound crystallizes in the space group I4(1)/amd (BaCd(11) structure type) with one Sr position (?overline{4}m2) and three Zn sites (?overline{4}m2, .2/m., 1). The structure is described in terms of all-face-capped Zn(8) tetrahedra as the central building unit, defined by the Zn atoms on .2/m. and 1. The building units are condensed into chains by the central tetrahedra sharing edges, and the chains are interconnected by shared capping atoms. The resulting three dimensional framework of Zn atoms yields channels that are occupied by Sr and Zn atoms on the high-symmetry ?overline{4}m2 positions. PMID- 17917210 TI - catena-Poly[[bis(nitrato-kappa(2)O,O')copper(II)]-mu-propane-1,3 diylbis(diphenylphosphine oxide)-kappa(2)O:O']. AB - The structure of the title complex, [Cu(NO(3))(2)(C(27)H(26)O(2)P(2))](n), consists of polymeric chains formed by propane-1,3-diylbis(diphenylphosphine oxide) ligands bridging between metal centres. The Cu atom lies on a twofold rotation axis and a further symmetry centre bisects the bridging bisphosphine dioxide ligand. The CuO(6) coordination geometry is a distorted octahedron, with the bidentate chelating nitrate groups adopting a cis configuration. PMID- 17917211 TI - 2,6-Bis(azaindole)pyridine: reactivity with iron(III) and copper(II) salts. AB - 1-[6-(1H-Pyrrolo[2,3-b]pyridin-1-yl)pyridin-2-yl]-1H-pyrrolo[2,3-b]pyridin-7-ium tetrachloridoferrate(III), (C(19)H(14)N(5))[FeCl(4)], (II), and [2,6-bis(1H pyrrolo[2,3-b]pyridin-1-yl-kappaN(7))pyridine-kappaN]bis(nitrato kappaO)copper(II), [Cu(NO(3))(2)(C(19)H(13)N(5))], (III), were prepared by self assembly from FeCl(3).6H(2)O or Cu(NO(3))(2).3H(2)O and 2,6-bis(1H-pyrrolo[2,3 b]pyridin-1-yl)pyridine [commonly called 2,6-bis(azaindole)pyridine, bap], C(19)H(13)N(5), (I). Compound (I) crystallizes with Z' = 2 in the P?overline{1} space group, with both independent molecules adopting a trans-trans conformation. Compound (II) is a salt complex with weak C-H...Cl interactions giving rise to a zigzag network with pi-stacking down the a axis. Complex (III) lies across a twofold rotation axis in the C2/c space group. The Cu(II) center in (III) has an N(3)O(2) trigonal-bipyramidal environment. The nitrate ligand coordinates in a monodentate fashion, while the bap ligand adopts a twisted tridentate binding mode. C-H...O interactions give rise to a ribbon motif. PMID- 17917212 TI - Hydrated hexacyanometallate(III) salts of triaqua(18-crown-6)lanthanoid(III) and tetraaqua(18-crown-6)lanthanoid(III) cations containing nine- and ten-coordinate lanthanoids. AB - Tetraaqua(18-crown-6)cerium(III) hexacyanoferrate(III) dihydrate, [Ce(C(12)H(24)O(6))(H(2)O)(4)][Fe(CN)(6)].2H(2)O, and tetraaqua(18-crown 6)neodymium(III) hexacyanoferrate(III) dihydrate, [Nd(C(12)H(24)O(6))(H(2)O)(4)][Fe(CN)(6)].2H(2)O, are isomorphous and isostructural in the C2/c space group, where the cations, which contain ten coordinate lanthanoid centres, lie across twofold rotation axes and the anions lie across inversion centres. In these compounds, an extensive series of O-H...O and O-H...N hydrogen bonds links the components into a continuous three dimensional framework. Triaqua(18-crown-6)lanthanoid(III) hexacyanoferrate(III) dihydrate, [Ln(C(12)H(24)O(6))(H(2)O)(3)][Fe(CN)(6)].2H(2)O, where Ln = Sm, Eu, Gd or Tb, are all isomorphous and isostructural in the P?overline{1} space group, as are triaqua(18-crown-6)gadolinium(III) hexacyanochromate(III) dihydrate, [Gd(C(12)H(24)O(6))(H(2)O)(3)][Cr(CN)(6)].2H(2)O, and triaqua(18-crown 6)gadolinium(III) hexacyanocobaltate(III) dihydrate, [Gd(C(12)H(24)O(6))(H(2)O)(3)][Co(CN)(6)].2H(2)O. In these compounds, there are two independent anions, both lying across inversion centres, and the lanthanoid centres exhibit nine-coordination; in the crystal structures, an extensive series of hydrogen bonds links the components into a three-dimensional framework. PMID- 17917213 TI - Bis(mu-4-carboxy-2-sulfonatobenzoato)bis[aqua(2,2'-bipyridyl)manganese(II)]. AB - In the title compound, [Mn(2)(C(8)H(4)O(7)S)(2)(C(10)H(8)N(2))(2)(H(2)O)(2)], pairs of hexacoordinated manganese(II) centres are bridged by 2 sulfonatoterephthalate(2-) anions to form cyclic centrosymmetric dimers, which are linked into sheets by O-H...O hydrogen bonds. PMID- 17917214 TI - Hydrogen bonding in diaquatetrakis(urea-kappaO)M(II) dinitrates, with M = Ni and Co. AB - The isomorphous title compounds, [Ni{(NH(2))(2)CO}(4)(H(2)O)(2)](NO(3))(2) and [Co{(NH(2))(2)CO}(4)(H(2)O)(2)](NO(3))(2), feature discrete centrosymmetric cations in octahedral coordinations, formed by four urea molecules linked via their O atoms to the central ion in equatorial positions and two water molecules in trans positions. The complexes are packed in a pseudo-hexagonal manner separated by the nitrate counter-ions. All H atoms are involved in moderate hydrogen bonds of four types: N-H...O=C, N-H...O-N, O-H...O-N and N-H...O-H. Graph-set analysis was applied to distinguish the hydrogen-bond patterns at unitary and higher level graph sets. PMID- 17917215 TI - A novel framework of N-H...Br hydrogen bonds forming Br(4,4'-bipyridinium)(4) supramolecular synthons: bis(4,4'-bipyridinium) tris[tetrabromidoferrate(III)] bromide. AB - In the asymmetric unit of the title compound, (C(10)H(10)N(2))(2)[FeBr(4)](3)Br, the Fe atoms are in a distorted tetrahedral environment. The crystal structure contains a novel arrangement of Br(4,4'-bipyridinium)(4) supramolecular synthons assembled via short N-H...Br hydrogen bonds (H...Br = 2.55, 2.40, 2.38 and 2.55 A), where four cations surround one nonbonded bromide ion in a tetrahedral arrangement. These synthons are further connected by hydrogen bonds using the remaining terminal NH hydrogens in each cation and the Br(-) ions to form an adamantoid-like network and thus produce a three-dimensional supramolecular architecture with the [FeBr(4)](-) ions located in the cavities. The structure shows no significant intermolecular Br...Br, Br...aryl or aryl-aryl interactions. PMID- 17917216 TI - Catena-poly[[dinitratocadmium(II)]bis[mu-bis(2-methyl-1H-imidazol-1-yl)methane kappa(2)N(3):N(3')]]. AB - The title compound, [Cd(NO(3))(2)(C(9)H(12)N(4))(2)](n), has a one-dimensional double-bridged chain polymer structure with a 16-membered macrometallacyclic tetragonal structural motif. The Cd(II) ion occupies a crystallographic inversion centre and is coordinated by four equatorial N atoms from four distinct bis(2 methylimidazol-1-yl)methane ligands and two apical nitrate O atoms to form a slightly distorted octahedral coordination geometry. PMID- 17917217 TI - [2,6-Bis(5-methyl-2-pyridyl)phenyl-kappa(3)N,C(1),N']chloridoplatinum(II). AB - In the title compound, [Pt(C(18)H(15)N(2))Cl], the Pt(II) centre adopts a distorted square-planar coordination geometry due to the pincer-type monoanionic N-C-N tridentate ligand. The planar complexes stack via pi-pi interactions to form two-dimensional accumulated sheets. This packing pattern is in contrast to that in related pincer-type N-C-N complexes, which exhibit a one-dimensional columnar stacking. PMID- 17917218 TI - Tetraaquabis(2-methoxybenzaldehyde isonicotinoylhydrazone)cadmium(II) dinitrate. AB - The Cd(II) centre in the title complex, [Cd(C(14)H(13)N(3)O(2))(2)(H(2)O)(4)](NO(3))(2), occupies a crystallographic inversion centre and is coordinated by two donor N atoms from two 2 methoxybenzldehyde isonicotinoylhydrazone ligands and by four O atoms from four coordinated water molecules, giving a slightly distorted octahedral geometry. There is an extended three-dimensional network structure resulting from O-H...O hydrogen bonds between coordinated water and nitrate anions, and between coordinated water and carbonyl O atoms, and from N-H...O hydrogen bonds between NH groups and nitrate O atoms. PMID- 17917219 TI - Aqua-2kappaO-mu-{N,N'-bis[2-(dimethylamino)ethyl]oxamidato(2-)} 1kappa(2)O,O':2kappa(4)N,N',N'',N'''-bis(4,4'-dimethyl-2,2'-bipyridine 1kappa(2)N,N')copper(II)nickel(II) bis(perchlorate) methanol hemisolvate. AB - In the crystal structure of the title complex, [CuNi(C(10)H(20)N(4)O(2))(C(12)H(8)N(2))(2)(H(2)O)](ClO(4))(2).0.5CH(4)O or [CuNi(dmaeoxd)(phen)(2)(H(2)O)](ClO(4))(2).0.5CH(3)OH {H(2)dmaeoxd is N,N'-bis[2 (dimethylamino)ethyl]oxamide and phen is 1,10-phenanthroline}, the deprotonated dmaeoxd(2-) ligand is in a cis conformation and bridges the Cu(II) and Ni(II) atoms. The Cu(II) atom is located in a slightly distorted square-based pyramidal environment, while the Ni(II) atom is in an irregular octahedral environment. The binuclear Cu-Ni units interact with each other via pi-pi interactions, which results in an extended chain along the b axis. PMID- 17917220 TI - cis-bis(dimethyl sulfide)dinitratoplatinum(II) at 295 and 150 K. AB - The title compound, [Pt(NO(3))(2)(C(2)H(6)S)(2)], crystallizes in the P2(1)/n space group (Z' = 2), with pseudo-square-planar coordination geometry. The complex forms dimers with pseudosymmetry C(i) arranged in columns along the b axis direction, with Pt...Pt distances of 6.3056 (3) and 4.2382 (2) A (at 150 K). Each column is surrounded by six other columns in a honeycomb rod-like packing. The coordination mode of the nitrate ligands is monodentate, with Pt-O-N angles ranging from approximately 117 to approximately 118 degrees and a tilt between the nitrate ligands and the coordination planes in the range approximately 63-70 degrees (at 150 K). The coordination mode of the nitrate ligands is compared with that observed in reported Pt(NO(3))(2)L(2) complexes (where L is a ligand with a donor atom from group 15 or 16), all of which are monodentate, with an average Pt O-N angle of 118 (2) degrees and a tilt in the range 90 +/- 30 degrees (with two exceptions, in which the nitrates are approximately in the coordination plane). PMID- 17917221 TI - Aqua(dipicolinato-kappa(3)O,N,O')(1H-imidazole-kappaN(3))(1,10-phenanthroline N,N')manganese(II). AB - In the title compound, [Mn(C(7)H(3)NO(4))(C(3)H(4)N(2))(C(12)H(8)N(2))(H(2)O)], the Mn(II) centre is surrounded by one bidentate phenanthroline ligand [Mn-N = 2.383 (3) and 2.421 (3) A], one tridentate dipicolinate ligand [Mn-N = 2.300 (3) A, and Mn-O = 2.300 (2) and 2.357 (2) A], one monodentate imidazole ligand [Mn-N = 2.238 (3) A] and one water molecule [Mn-O = 2.157 (3) A]. It displays a distorted pentagonal-bipyramidal geometry, with neighbouring angles within the equatorial plane ranging from 68.05 (9) to 77.48 (10) degrees . Intermolecular O H...O hydrogen bonds link the molecules into infinite chains. The chains are crosslinked by hydrogen bonds involving the carboxyl O atoms of the dipicolinate ligand and the protonated imidazole N atom, leading to an infinite two dimensional network sheet packing mode. The complete solid-state structure can be described as a three-dimensional supramolecular framework, stabilized by these intermolecular hydrogen-bonding interactions and pi-pi stacking interactions involving the phenanthroline rings. PMID- 17917222 TI - Two substituted 2-pyrrolin-5-ones: chains built from a single N-H...O hydrogen bond. AB - In each of methyl 2-methyl-5-oxo-2-pyrroline-3-carboxylate, C(7)H(9)NO(3), and 3 acetyl-2-methyl-2-pyrrolin-5-one, C(7)H(9)NO(2), the pyrrolinone ring is planar. In each structure, molecules are linked into simple chains by way of a single N H...O hydrogen bond. PMID- 17917223 TI - pi-Stacked dimers in 6-methoxy-3,3-dimethyl-3H-benzo[f]chromene, and centrosymmetric dimers containing C-H...pi(arene) hydrogen bonds in racemic 3 bromo-2,2,6,6-tetramethyl-3,4-dihydro-2H,6H-1,5-dioxatriphenylene. AB - The title compounds, namely 6-methoxy-3,3-dimethyl-3H-benzo[f]chromene, C(16)H(16)O(2), (III), and racemic 3-bromo-2,2,6,6-tetramethyl-3,4-dihydro-2H,6H 1,5-dioxatriphenylene, C(20)H(21)BrO(2), (IV), were both synthesized in one-step regioselective Wittig reactions from substituted 1,2-naphthoquinones. The new ring in both compounds adopts a screw-boat conformation. A single pi-pi stacking interaction links the molecules of (III) into centrosymmetric dimeric aggregates, and a single C-H...pi(arene) hydrogen bond links the molecules of (IV) into centrosymmetric dimers. PMID- 17917224 TI - N-acetyl-L-tyrosine methyl ester monohydrate at 293 and 123 K. AB - The crystal structure of a protected L-tyrosine, namely N-acetyl-L-tyrosine methyl ester monohydrate, C(12)H(15)NO(4).H(2)O, was determined at both 293 (2) and 123 (2) K. The structure exhibits a network of O-H...O and N-H...O hydrogen bonds, in which the water molecule plays a crucial role as an acceptor of one and a donor of two hydrogen bonds. Molecules of water and of the protected L-tyrosine form hydrogen-bonded layers perpendicular to [001]. C-H...pi interactions are observed in the hydrophobic regions of the structure. The structure is similar to that of N-acetyl-L-tyrosine ethyl ester monohydrate [Soriano-Garcia (1993). Acta Cryst. C49, 96-97]. PMID- 17917225 TI - 2,3,10,11-Tetrathiatricyclo[10.4.0.0(4,9)]hexadeca-4,6,8,12,14,16-hexaene (precession camera versus CCD). AB - The title compound, C(12)H(8)S(4), has crystallographic ?overline{1} symmetry, the benzene groups thus being anti with respect to the plane of the four S atoms. The S-S and C-S bond lengths of the sulfur-carbon eight-membered ring were found to be similar to those in other structures containing such sulfur-carbon rings. There is evidence for pi-pi interactions between the aromatic rings of neighboring molecules, linking them into sheets. PMID- 17917226 TI - 3-Deoxy-beta-D-ribo-hexopyranose (3-deoxy-beta-D-glucopyranose). AB - The beta-pyranose form, (III), of 3-deoxy-D-ribo-hexose (3-deoxy-D-glucose), C(6)H(12)O(5), crystallizes from water at 298 K in a slightly distorted (4)C(1) chair conformation. Structural analyses of (III), beta-D-glucopyranose, (IV), and 2-deoxy-beta-D-arabino-hexopyranose (2-deoxy-beta-D-glucopyranose), (V), show significantly different C-O bond torsions involving the anomeric carbon, with the H-C-O-H torsion angle approaching an eclipsed conformation in (III) (-10.9 degrees ) compared with 32.8 and 32.5 degrees in (IV) and (V), respectively. Ring carbon deoxygenation significantly affects the endo- and exocyclic C-C and C-O bond lengths throughout the pyranose ring, with longer bonds generally observed in the monodeoxygenated species (III) and (V) compared with (IV). These structural changes are attributed to differences in exocyclic C-O bond conformations and/or hydrogen-bonding patterns superimposed on the direct (intrinsic) effect of monodeoxygenation. The exocyclic hydroxymethyl conformation in (III) (gt) differs from that observed in (IV) and (V) (gg). PMID- 17917228 TI - Hydrogen-bonded sheets in benzylmethylammonium hydrogen maleate. AB - In the title compound, C(8)H(12)N(+).C(4)H(3)O(4)(-), there is a short and almost linear but asymmetric O-H...O hydrogen bond in the anion. The ions are linked into C(2)(2)(6) chains by two short and nearly linear N-H...O hydrogen bonds and the chains are further weakly linked into sheets by a single C-H...O hydrogen bond. PMID- 17917227 TI - 8-tert-Butyl-7-(4-chlorophenyl)-10-phenyl-5,6-dihydro-10H-benzo[h]pyrazolo[3,4 b]quinoline. AB - The title compound, C(30)H(26)ClN(3), crystallizes with Z' = 3 in the P?overline{1} space group. The three independent molecules have very similar, although not identical, conformations, with almost perfect screw-boat forms adopted by the non-aromatic carbocyclic rings. Four independent C-H...pi(arene) hydrogen bonds link the molecules into centrosymmetric six-component aggregates. PMID- 17917229 TI - A new polymorph of tetraphenyldiboroxane. AB - A new polymorph of tetraphenyldiboroxane [or oxybis(diphenylborane)], C(24)H(20)B(2)O, (Ia), has been found. It is monoclinic, like the already known form, (Ib), and can be refined in the same space group, namely P2(1)/c, or in the equivalent setting P2(1)/n. The molecular conformations of the two polymorphs differ in the rotations of two of the phenyl rings about the B-C bonds, leading to markedly different packing patterns and cell dimensions. PMID- 17917230 TI - Hexagonal packing of cis,cis-cyclohexane-1,3,5-tricarboxamide. AB - The title compound, C(9)H(15)N(3)O(3), which has crystallographically imposed threefold symmetry, crystallizes as a hexagonal columnar structure. The crystal structure is stabilized by a less common amide-amide synthon, where one amide group is hydrogen bonded to four others. The amide groups form cyclic amide-amide hexamers via N-H...O hydrogen bonds. PMID- 17917231 TI - A new conformer of 1,4,7-tris(p-tolylsulfonyl)-1,4,7-triazacyclononane. AB - A second polymorphic form (form II) of the previously reported 1,4,7-tris(p tolylsulfonyl)-1,4,7-triazacyclononane (form I), C(27)H(33)N(3)O(6)S(3), is presented. The molecular structures of the two forms display very different conformations, thus prompting the two forms to crystallize in two different space groups and exhibit quite diverse crystal structure assemblies. Form I crystallizes in the triclinic space group P?overline{1}, while form II crystallizes in the monoclinic space group P2(1)/n. The main differences between the two molecular structures are the conformations of the p-tosyl groups relative to each other and to the macrocyclic ring. The resulting crystal packing displays no classical hydrogen bonds, but different supramolecular synthons give rise to different packing motifs. PMID- 17917232 TI - Acyclic Schiff base salts derived from 1,3-diaminopropane and 1,3-diamino-2 hydroxypropane. AB - Two salts of acyclic Schiff base cationic ligands, namely N,N'-bis(2 nitrobenzyl)propane-1,3-diammonium dichloride monohydrate, C(17)H(22)N(4)O(4)(2+).2Cl(-).H(2)O, (I), and 2-hydroxy-N,N'-bis(2 nitrobenzyl)propane-1,3-diammonium dichloride, C(17)H(22)N(4)O(5)(2+).2Cl(-), (II), were synthesized as precursors in order to obtain new acyclic and macrocyclic multidentate ligands and complexes. The cation conformations in compounds (I) and (II) are different in the solid state, although the cations are closely related chemically. Similarly, the hydrogen-bonding networks involving ammonium cations, hydroxyl groups and chloride anions are also different. In the cation of compound (II), the hydroxyl group is disordered over two sets of sites, with occupancies of 0.785 (8) and 0.215 (8). PMID- 17917233 TI - The 7-bromo derivative of 2-amino-2'-deoxytubercidin fluorinated at the sugar moiety. AB - The title compound, 2,4-diamino-5-bromo-7-(2-deoxy-2-fluoro-beta-D arabinofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine, C(11)H(13)BrFN(5)O(3), shows two conformations of the exocyclic C4'-C5' bond, with the torsion angle gamma (O5' C5'-C4'-C3') being 170.1 (3) degrees for conformer 1 (occupancy 0.69) and 60.7 (7) degrees for conformer 2 (occupancy 0.31). The N-glycosylic bond exhibits an anti conformation, with chi = -114.8 (4) degrees . The sugar pucker is N-type (C3'-endo; (3)T(4)), with P = 23.3 (4) degrees and tau(m) = 36.5 (2) degrees . The compound forms a three-dimensional network that is stabilized by several intermolecular hydrogen bonds (N-H...O, O-H...N and N-H...Br). PMID- 17917234 TI - 2,3,5,6-Tetrakis[3,5-bis(trifluoromethyl)phenoxy]-2,5-bis(dimethylamino)2,3,5,6 tetrabora-1,4-dioxane diethyl ether 0.667-solvate. AB - The title compound, C(36)H(26)B(4)F(24)N(2)O(6).0.667C(4)H(10)O, has centrosymmetric tetraboradioxane molecules, half each of three of these comprising the asymmetric unit together with a molecule of diethyl ether. Disorder affects most of the CF(3) groups and one ethyl group of the solvent molecule. The B(4)O(2) rings are approximately planar and contain two B atoms with trigonal geometry and two with distorted tetrahedral geometry, the B-O bonds for the four-coordinate B atoms being longer than those for the three-coordinate B atoms. N-H...O hydrogen bonds link two of the crystallographically independent molecules together in chains, while the third molecule forms discrete trimolecular clusters with two solvent molecules via N-H...O hydrogen bonds. This is the first crystallographically characterized example of a tetrabora-dioxane molecule containing both four- and three-coordinate B atoms. PMID- 17917235 TI - 5-(5-Nitropyridin-2-yldithio)-1-phenyl-1H-tetrazole. AB - In the title disulfide, C(12)H(8)N(6)O(2)S(2), all bond lengths and angles are within normal ranges, and the molecules are linked into centrosymmetric R(2)(2)(20) dimers by simple C-H...N interactions. Weak intermolecular C H...pi(arene) and pi-pi interactions, involving the benzene CH groups and the benzene rings, and the pyridine rings, respectively, further stabilize and reinforce the crystal structure. PMID- 17917236 TI - Ammonium violurate: a compact structure with extensive hydrogen bonding in three dimensions. AB - The title compound [systematic name: ammonium pyrimidine-2,4-5,6(1H,3H)-tetrone 5 oximate], NH(4)(+).C(4)H(2)N(3)O(4)(-), crystallizes from water in the triclinic space group P?overline{1} and is ismorphous with a known rubidium complex [Gillier (1965). Bull. Soc. Chim. Fr. pp. 2373-2384]. The principal feature of the structure is hydrogen bonding; each ammonium H atom acts as a bifurcated donor and three of the four violurate O atoms are bifurcated acceptors, with the fourth acting as a trifurcated acceptor. The pattern of hydrogen bonding around the cation is very similar to the rubidium coordination environment in the related structure. The violurate anions pack as hydrogen-bonded crinkled tapes, which are linked and separated by the ammonium cations to give a compact three dimensional structure. PMID- 17917237 TI - Trierixin, a novel Inhibitor of ER stress-induced XBP1 activation from Streptomyces sp. 1. Taxonomy, fermentation, isolation and biological activities. AB - In the course of screening for an inhibitor of ER stress-induced XBP1 activation, we isolated a new member of the triene-ansamycin group compound, trierixin, from a culture broth of Streptomyces sp. AC 654. Trierixin was purified by column chromatography on silica gel and by HPLC. The molecular formula of trierixin is C(37)H(52)N(2)O(8)S. Trierixin inhibited thapsigargin-induced XBP1-luciferase activation in HeLa/XBP1-luc cells and endogenous XBP1 splicing in HeLa cells with an IC(50) of 14 ng/ml and 19 ng/ml, respectively. Moreover, in the process of isolating trierixin, we isolated structurally related mycotrienin II and trienomycin A as inhibitors of ER stress-induced XBP1 activation from a culture broth of a trierixin-producing strain. This study provides the first observation that triene-ansamycins have a novel inhibitory effect against XBP1 activation. PMID- 17917238 TI - Isolation and structure elucidation of thiazomycin- a potent thiazolyl peptide antibiotic from Amycolatopsis fastidiosa. AB - Thiazolyl peptides are a class of rigid macrocyclic compounds richly populated with thiazole rings. They are highly potent antibiotics but none have been advanced to clinic due to poor aqueous solubility. Recent progress in this field prompted a reinvestigation leading to the isolation of a new thiazolyl peptide, thiazomycin, a congener of nocathiacins. Thiazomycin possesses an oxazolidine ring as part of the amino-sugar moiety in contrast to the dimethyl amino group present in nocathiacin I. The presence of the oxazolidine ring provides additional opportunities for chemical modifications that are not possible with other nocathiacins. Thiazomycin is extremely potent against Gram-positive bacteria both in vitro and in vivo. The titer of thiazomycin in the fermentation broth was very low compared to the nocathiacins I and III. The lower titer together with its sandwiched order of elution presented significant challenges in large scale purification of thiazomycin. This problem was resolved by the development of an innovative preferential protonation based one- and/or two-step chromatographic method, which was used for pilot plant scale purifications of thiazomycin. The isolation and structure elucidation of thiazomycin is herein described. PMID- 17917239 TI - Antibacterial evaluations of thiazomycin- a potent thiazolyl peptide antibiotic from Amycolatopsis fastidiosa. AB - Thiazomycin is a novel thiazolyl peptide closely related to nocathiacin I. It was isolated from Amycolatopsis fastidiosa by chemical and biological screening. Thiazomycin showed highly potent bactericidal activity against Gram-positive pathogens (MIC range 0.002 approximately 0.064 microg/ml) and did not show cross resistance to clinically relevant antibiotic classes such as beta-lactams, vancomycin, oxazolidinone and quinolones. It was highly efficacious against Staphylococcus aureus infection in mice exhibiting an ED(99) value of 0.15 mg/kg by subcutaneous administration. It inhibited bacterial growth by selective inhibition of protein synthesis and it was thought to interact with L11 protein and 23S rRNA of the 50S ribosome. Structurally, it possesses an oxazolidine ring in the amino-sugar residue that provides further opportunities for selective chemical modifications that are not feasible with other thiazolyl peptides. More importantly such a modification can potentially lead to semi-synthetic compounds that overcome problems that have hampered clinical development of this class of compounds. Despite its positive attributes, emergence of an unacceptable frequency of resistance poses significant challenges for further development of thiazomycin and this class of molecules for therapeutic use. PMID- 17917240 TI - In vitro activity of novel rifamycins against gram-positive clinical isolates. AB - We describe novel rifamycins that have improved activity, compared with rifampin, against clinical isolates of staphylococci and streptococci, with MIC(90)s of 0.008 and 0.0005 microg/ml, respectively. This enhanced antibacterial activity, along with their potential lack of drug-drug interactions, are considerations that suggest the potential of these novel rifamycins in combination therapy to treat serious Gram-positive infections. PMID- 17917241 TI - Comparison of the bioactive secondary metabolites from the scale insect pathogens, Anamorph Paecilomyces cinnamomeus, and Teleomorph Torrubiella luteorostrata. AB - A scale insect pathogen Paecilomyces cinnamomeus BCC 9616 and its teleomorph Torrubiella luteorostrata BCC 9617, collected on the same host specimen, were fermented and chemically explored. Both fungi produced paecilodepsipeptide A (1) and zeorin (4) as major constituents of mycelia extracts. The culture broth extract of BCC 9616 provided a known diketopiperazine, terezine D (5), and a new xanthone glycoside, norlichexanthone-6-O-beta-(4-O-methylglucopyranoside) (6). On the other hand, the broth extract of BCC 9617 contained small amounts of a new naphthopyrone glycoside, rubrofusarin-6-O-beta-(4-O-methylglucopyranoside) (7) along with 5. Structures of the new compounds, 6 and 7, were elucidated by interpretation of NMR and mass spectroscopic data. The overall results demonstrated that the metabolite profiles of the cultured anamorph (BCC 9616) and teleomorph (BCC 9617) originating from the same host specimen resemble each other closely. The (1)H-NMR spectroscopic analysis of the culture extracts from other strains of P. cinnamomeus and T. luteorostrata revealed that zeorin is the most commonly occurring fermentation product of these fungi, whereas paecilodepsipeptide A was the metabolite specific to the particular isolate BCC 9616/BCC 9617. PMID- 17917243 TI - Prenylterphenyllin and its dehydroxyl analogs, new cytotoxic substances from a marine-derived Fungus Aspergillus candidus IF10. AB - Three novel cytotoxic substances named prenylterphenyllin (1), 4'' deoxyprenylterphenyllin (2), and 4''-deoxyisoterprenin (3) were isolated from a cultured marine-derived fungus of Aspergillus candidus IF10 together with 4'' deoxyterprenin (4). Their chemical structures were elucidated on the basis of 2D NMR analysis. These compounds 1 approximately 4 showed cytotoxic activity against human epidermoid carcinoma KB cells (KB3-1) with IC(50) of 8.5, 3.0, 2.5, and 4.5 microg/ml, respectively. PMID- 17917242 TI - Trierixin, a novel Inhibitor of ER stress-induced XBP1 activation from Streptomyces sp. II. structure elucidation. AB - Trierixin, a new member of the triene-ansamycin group, has been isolated from the fermentation broth of Streptomyces sp. AC654 as an inhibitor of ER stress-induced XBP1 activation. The structure of trierixin was determined on the basis of its spectroscopical and chemical properties. Trierixin possessed a 21-membered macrocyclic lactam, which contains a methylthio-benzenediol structure, and a cyclohexanecarbonylalanine moiety. Trierixin is thus elucidated as 21 thiomethylmycotrienin II. PMID- 17917244 TI - The Fgf families in humans, mice, and zebrafish: their evolutional processes and roles in development, metabolism, and disease. AB - Fibroblast growth factors (Fgfs) were originally isolated as growth factors for fibroblasts. However, Fgfs are now recognized as polypeptide growth factors of ca. 150-250 amino acid residues with diverse biological activities and expression profiles. The Fgf signaling system has been identified in multicelluar but not in unicellular organisms. In contrast to the only two Fgf genes and one Fgf receptor (Fgfr) gene in Caenorhabditis elegans, both the human and mouse Fgf and Fgfr gene families comprise twenty-two and four members, respectively. Their evolutional processes indicate that the Fgf and Fgfr gene families greatly co-expanded during the evolution of early vertebrates. The expansion of the Fgf and Fgfr gene families has enabled this signaling system to acquire diversity of function and a nearly ubiquitous involvement in many developmental and physiological processes. The zebrafish fgf gene family comprises twenty-seven members with several paralogs generated by an additional genome duplication. The mouse and zebrafish are useful models for studying gene functions. Fgf knockout mice have been generated. Several Fgf knockout mice die in the embryonic or early postnatal stages, indicating crucial roles for these genes in various developmental processes. However, other Fgf knockout mice survive with subtle phenotypic alterations. Their functions might be redundant. Studies using zebrafish embryos with mutated or knockdown fgfs also indicate that fgfs play crucial roles in development in that species. Although most Fgfs act in development in a paracrine and/or autocrine manner, some have potential roles in metabolism in an endocrine manner. In humans, Fgf signaling disorders result in hereditary diseases and cancers. PMID- 17917245 TI - Production and regulation of eotaxin-2/CCL24 in a differentiated human leukemic cell line, HT93. AB - When a human leukemic cell line, HT93 was incubated with all-trans retinoic acid (ATRA), IL-5, or both, this cell line was differentiated into eosinophic lineage, in that an eosinophilic specific granule proteins, major basic protein (MBP) and eosinophil peroxidase (EPO) appeared. Both CD11b and CC chemokine receptor, CCR3 expression were upregulated, while CD71 expression was downregulated by ATRA or ATRA+IL-5. Concomitantly, marked production of eotaxin-2/CCL24 was observed, but no production of eotaxin-1/CCL11 and eotaxin-3/CCL26 was detected. Since only 20 to 30% cells incubated with ATRA became positive for CCR3, CCR3(+) population was enriched by a magnetic activated cell sorter (MACS). Enriched CCR3(+) population produced higher eotaxin-2/CCL24 than the CCR3(-) population, indicating that differentiated eosinophils are capable of producing eotaxin-2/CCL24. During the ATRA-induced differentiation, expression of a transcriptional factor, GATA-1 was significantly increased. Introduction of siRNA against GATA-1 markedly reduced the ATRA-induced differentiation markers including CD11b and CCR3, as well as reduced eotaxin-2/CCL24 production. Finally, ATRA-induced differentiation and eotaxin-2/CCL24 production were greatly enhanced in the GATA-1-overexpressed clones. These results indicate that the ability to produce eotaxin-2/CCL24 is acquired during the differentiation into eosinophilic lineage which is dependent on GATA-1 expression. PMID- 17917246 TI - Glycated human serum albumin induces interleukin 8 mRNA expression through reactive oxygen species and NADPH oxidase-dependent pathway in monocyte-derived U937 cells. AB - Glycated human serum albumin (Glc-HSA) has previously been reported (Higai K., et al., 2006) to induce E-selectin expression on human umbilical vein endothelial cells through activation of NADPH oxidase; however, Glc-HSA signaling in monocytes remains obscure. To clarify the influence on human monocyte-derived U937 cells, U937 cells were stimulated with Glc-HSA and glycoaldehyde-dimer modified HSA (GA-HSA) for 2 h in the absence and presence of the protein kinase C (PKC) inhibitor calphostin and the reactive oxygen species (ROS) scavenger N acetyl-L-cysteine (NAC) and NADPH oxidase inhibitor apocynin; interleukin-8 (IL 8) mRNA expression was determined by RT-PCR. As a result, IL-8 mRNA expression in U-937 cells was time- and dose-dependently enhanced by stimulation with Glc-HSA and GA-HSA. Furthermore, promoter activity of the IL-8 reporter gene was enhanced approximately 2-fold by stimulation with Glc-HSA and GA-HSA. Nuclear factor kappaB (NFkappaB) and activator protein-1 (AP-1) reporter genes were also enhanced although CCAAT/enhancer binding protein (C/EBP) was not affected. IL-8 mRNA expression was suppressed by NAC and apocynin but not calphostin in cells stimulated with Glc-HSA; however, its expression in cells stimulated with GA-HSA was suppressed by calphostin but not NAC. These results indicated that IL-8 mRNA expression was upregulated by NFkappaB and AP-1 in U937 cells stimulated with Glc HSA and GA-HSA, but the signaling pathways were different. PMID- 17917247 TI - Amine modification of digested peptide at C-terminal end during protein digestion by protease. AB - We recently reported that C-terminal polyamine modification occurs when proteins are digested with trypsin in the presence of polyamine [Biochem. Biophys. Res. Commun., 356, 159-162 (2007)]. In the present study, the characteristics of this C-terminal modification in the presence of protease and amine were investigated. When hemoglobin (HB) was digested with trypsin in the presence of N-(2-pyridyl) 1,4-diaminobutane (Py4), formation of the modified peptide was dependent on time and on HB or Py4 concentration. When synthetic peptide was treated with trypsin in the presence of Py4, ca. 0.1% of the peptide was modified with Py4. When HB or cytochrome C was treated with a range of serine proteases in the presence of various amines (Py4, N-(2-pyridyl)-1,3-diaminopropane, tranexamic acid, isonicotinic acid hydrazide and ampicillin), the modified peptide was detected in all cases tested, thus suggesting that amine modification widely accompanies digestion by proteases. PMID- 17917248 TI - Establishment of evaluation method for siRNA delivery using stable cell line carrying the luciferase reporter gene. AB - We determined the influence of siRNA (short interfering RNA) for expression of plasmid DNA (pDNA), when mismatched siRNA and pDNA encoding beta-galactosidase (beta-gal) were transfected into HeLa cells by the cotransfection method in which they were simultaneously added to the cells. Cationic liposomes (Lipofectamine2000) were used as a gene transfection reagent. The knockdown effect on beta-gal was observed even when mismatched siRNA was used, and the effect depended on the amount of added mismatched siRNA. But, there was not a distinct difference of introduction of pDNA into cells between using mismatched siRNA and without using it. We considered that the cotransfection method should be avoided when we confirm RNAi efficiency. The reliable evaluation method for siRNA delivery in vitro was thus established by using NFAT reporter HeLa stable cell line or CHO (pMAM-luc) cell line that had DNA encoding luciferase. The following experimental conditions for each cell line were optimized: cell numbers seeded, total incubation times, concentrations of added inducers, and incubation times after addition of inducers. Transfection performance was compared for six commercially available reagents by this method. No commercially available transfection reagent, however, could reduce luciferase activity by less than one tenth without causing cellular cytotoxicity. Development of novel reagents providing higher transfection effects without cytotoxicity is needed. PMID- 17917249 TI - Cloning and functional expression of ubiquitin-like protein specific proteases genes from Candida albicans. AB - The small ubiquitin-like modifier (SUMO) modification occurred at bud necks and sites of septum formation in hyphae of the opportunistic fungal pathogen Candida albicans. Three genes encoding putative SUMO deconjugation enzymes (Ulp, ubiquitin-like protein specific proteases) of C. albicans were obtained through sequence database searching with Ulp domain of Saccharomyces cerevisiae Ulp1 (ScUlp1). These genes were designated as CaULP1, CaULP2 and CaULP3. The open reading frames of three putative ULPs were cloned and expressed in Pichia pastoris, resulting recombinant proteins. Functional analysis of recombinant CaUlp1, CaUlp2 and CaUlp3 confirms that these proteins exhibit SUMO-processing activity. CaULP1, CaULP2 and CaULP3 only expressed active form enzyme in P. pastoris but not in Escherichia coli. The molecular weights of CaUlp1, CaUlp2 and CaUlp3 proteins expressed in P. pastoris were larger than theoretical molecular weights. This observation was in good agreement with result of Western blot analysis of CaUlp1 and CaUlp3 proteins in C. albicans. It was assumed that CaUlp1, CaUlp2 and CaUlp3 proteins may need post-translational modifications to exhibit SUMO-processing activity. To our knowledge, this is the first report on cloning and expression of Ulp genes from C. albicans. Furthermore RT-PCR and Western blot analysis show that CaULP2 has no detectable expression both in yeast and in hyphal forms of C. albicans. PMID- 17917250 TI - Blood cell and vessel formation following transplantation of activin-treated explants in Xenopus. AB - Treatment of Xenopus blastula with activin converts undifferentiated presumptive ectoderm (animal cap) into mesoderm and endoderm in a dose-dependent manner. At low concentrations, activin induces ventral mesoderm such as blood cells. Here we show that activin-treated aggregates of animal cap cells prepared from undifferentiated presumptive ectoderm and transplanted into Xenopus embryos differentiated to form red blood cells and vascular endothelial cells. We compared gene expression profiles of the activin-treated with untreated aggregates of animal cap cells using microarray analysis. This revealed 838 clones including vascular-related genes that were expressed at levels at least 2 fold greater in the activin-treated aggregates than in the untreated controls. Of these, 356 were known Xenopus genes, 296 had homologues, and 186 were unknown genes. These findings identified novel vascular-related genes and provided insights into how the blood vessel system establishes in normal development. PMID- 17917252 TI - In vitro antimicrobial activity and the mode of action of indole-3-carbinol against human pathogenic microorganisms. AB - Indole-3-carbinol (I3C) is a naturally occurring constituent of cruciferous vegetables. The aim of this study was to assess the in vitro antimicrobial activity of I3C and its mode of action. By using an NCCLS broth microdilution assay, the activity of I3C was evaluated against human pathogenic microorganisms including clinically isolated antibiotic-resistant bacterial strains. The results indicated that I3C exhibited broad spectrum antimicrobial activities. To elucidate the physiological changes of the fungal cells induced by I3C, we performed a flow cytometric analysis for a cell cycle. The results showed that I3C arrested the cell cycle at the G(2)/M phase in Candida albicans. To understand the antifungal mode of action of I3C, the change in the membrane dynamics was monitored by using fluorescence changing experiments against C. albicans. The results suggest that I3C may exert antifungal activity by disrupting the structure of the cell membrane. The present study indicates that I3C has considerable antimicrobial activity, deserving further investigation for clinical applications. PMID- 17917251 TI - LIF- and IL-6-induced acetylation of STAT3 at Lys-685 through PI3K/Akt activation. AB - Signal transducer and activator of transcription 3 (STAT3), which mediates biological actions in many physiological processes, is activated by cytokines and growth factors via specific tyrosine or serine phosphorylation, dimerization and nuclear translocation. A recent study has demonstrated, by using antibody to acetylated lysine, and a STAT3 mutant with Lys-685-to-Arg substitution, that STAT3 is acetylated at Lys-685 by histone acetyltransferase p300, and that acetylation at Lys-685 is critical for STAT3 activation. In the present study, we created an acetyl-specific antibody against STAT3 acetylated at Lys-685, and found that leukemia inhibitory factor (LIF) or interleukin (IL)-6 induced acetylation of STAT3 at Lys-685 in 293T and Hep3B cells. Moreover, acetylation of STAT3 at Lys-685 was suppressed by PI3K inhibitor LY294002, or a dominant negative Akt. Taken together, our findings demonstrate that endogenous STAT3 is acetylated at Lys-685 by LIF or IL-6 through PI3K/Akt activation. PMID- 17917253 TI - Comparison of pathogenicity of various Candida albicans and C. stellatoidea strains. AB - In order to clarify the pathogenicity and the pathogenic factors of various Candida species strains, three strains, NIH A-207 and J-1012 (serotype A), and NIH B-792 (serotype B) of Candida albicans and two strains, ATCC 20408 (karyotype II) and ATCC 36232 (karyotype I) of C. stellatoidea, a synonym for C. albicans, were tested for their lethality to mice, adherence to Hela cells, hydrophobicity, and cell growth under acidic conditions, pH 2.0-5.9. The pathogenicity for mice of all the strains was observed in the order NIH B-792, ATCC 36232, J-1012, NIH A 207, and ATCC 20408. The pathogenicity for mice by all the strains used was well correlated with adherence to the Hela cells, the hydrophobicity, and the cell growth under the acidic condition, pH 2.0. These results emphasize that these specific properties of the C. albicans and C. stellatoidea strains play an important role in the pathogenesis of candidosis. PMID- 17917254 TI - Experimental model of lower limb ischemia in rats and the effect of YM466, an oral direct factor Xa inhibitor. AB - A simple, quantitative, and reproducible model of lower limb ischemia was developed. Vascular injury was induced by ferric chloride (FeCl(3)) solution to the rat iliac artery, after which blood flow in all of the lower limbs were continuously monitored using a scanning laser Doppler blood flowmeter. After FeCl(3) injury, a distinct decrease in blood flow in the ischemic lower limb was observed and blood flow did not recover during the 30 min after vascular injury. YM466, an oral direct factor Xa inhibitor, dose-dependently inhibited the reduction of peripheral blood flow. The area under the blood flow-time curve during 30 min after vascular injury improved dose-dependently, with significance at doses of 3 and 10 mg/kg. These results suggest that factor Xa inhibitors are effective in patients with peripheral arterial disease, and that this vascular injury model is a useful tool for the screening and evaluation of the efficacy of new antithrombotic agents. PMID- 17917255 TI - Isobavachalcone, a chalcone constituent of Angelica keiskei, induces apoptosis in neuroblastoma. AB - Six chalcones from Angelica keiskei KOIDZUMI (Ashitaba in Japanese) and two chalcones from Humulus lupulus L. (hop) were examined for their cytotoxicity in two human neuroblastoma cell lines (IMR-32 and NB-39) and normal cells (primary culture of rat cerebellar granule cells) by [3-(4,5)-dimethyl-2-thiazolyl]-2,5 diphenyl-2H-tetrazolium bromide (MTT) assay. All chalcones exhibited cytotoxicity against neuroblastoma cells, and two of them (isobavachalcone and xanthoangelol H) had no effect on normal cells even at high concentration (10(-4) M) exposure. Typical morphologic features of apoptosis, including cell shrinkage, chromatin condensation, nuclear fragmentation and formation of apoptotic bodies, were observed in isobavachalcone-treated cells by Hoechst 33342 staining. Western blot analysis showed that isobavachalcone significantly reduced pro-caspase-3 and pro caspase-9, and subsequently increased the level of cleaved caspase-3 and cleaved caspase-9 in both neuroblastoma cell lines. Moreover, Bax was markedly induced by isobavachalcone application. These results suggest that isobavachalcone induces apoptotic cell death in neuroblastoma via the mitochondrial pathway and has no cytotoxicity against normal cells. Therefore, isobavachalcone may be applicable as an efficacious and safe drug for the treatment of neuroblastoma. PMID- 17917256 TI - Component and antioxidant properties of polysaccharide fractions isolated from Angelica sinensis (OLIV.) DIELS. AB - An analytical method of high performance capillary electrophoresis (HPCE) was developed to simultaneously separate and identify the component monosaccharides of Angelica sinensis polysaccharide fractions (APFs), named APF1, APF2 and APF3. The predominant sugars in APFs were identified as arabinose, glucose, rhamnose, galactose and galacturonic acid as well as trace amount of mannose and glucuronic acid, and the fractionation altered significantly the distribution of component monosaccharides in APFs. APF3 was the most active fraction to effectively inhibit H(2)O(2)-caused decrease of cell viability, lactate dehydrogenase (LDH) leakage and malondialdehyde (MDA) formation, and also reduced H(2)O(2)-caused decline of superoxide dismutase (SOD) activity and glutathione (GSH) depletion (p<0.05), followed by APF2 and APF1 in decreasing order. Furthermore, it was found that APFs (100 microg/ml) could protect macrophages by inhibiting the release of excess NO and reactive oxygen species (ROS) induced by high concentrations of H(2)O(2) (0.8-1.6 mM). PMID- 17917257 TI - Asp125 and Thr130 in transmembrane domain 3 are major sites of alpha1b-adrenergic receptor antagonist binding. AB - Site-directed mutagenesis was used to investigate the molecular interactions involved in prazosin binding to the human alpha(1b)-adrenergic receptor (alpha(1b)-AR) receptor. Based on molecular modeling studies, Thr130 and Asp125 in transmembrane region III of the alpha(1b)-AR receptor were found to interact with prazosin. Thr130 and Asp125 were mutated to alanine (Ala) and expressed in HEK293 cells. The radioligand [(3)H]prazosin did not show any binding to Asp125Ala mutant of alpha(1b)-AR. Therefore, it was not possible to find any prazosin affinity to the mutant using the radioligand [(3)H]prazosin. The mutation also abolished phenylephrine-stimulated inositol phosphate (IP) formation of [(3)H]myo-inositol. On the other hand, the Thr130Ala mutant showed reduced binding affinity for [(3)H]prazosin (dissociation constant, K(d) 674.27 pM versus 90.27 pM for the wild-type receptor) and had reduced affinity for both tamsulosin and prazosin (11-fold and 9-fold, respectively). However, the Thr130Ala mutant receptor retained the ability to stimulate the formation of [(3)H]myo-inositol. The results provide direct evidence that Asp125 and Thr130 are responsible for the interactions between alpha(1b)-AR receptor and radioligand [(3)H]prazosin as well as tamsulosin. PMID- 17917258 TI - Effects of urocortin, corticotropin-releasing factor (CRF) receptor agonist, and astressin, CRF receptor antagonist, on the sleep-wake pattern: analysis by radiotelemetry in conscious rats. AB - Stress has been known to release corticotropin-releasing factor (CRF) and have an affect on sleep-wake patterns. However, there is no direct evidence of CRF receptor agonist and antagonist on sleep-wake patterns. Therefore, this study aimed to clarify this point by using radiotelemetry system in conscious rats. Wake, non-rapid eye-moving (NREM) sleep and rapid eye-moving (REM) sleep were analyzed by computer software, simultaneously measuring electroencephalogram and electromyogram. In the light period, urocortin (CRF receptor agonist: i.v.) significantly increased wake duration, and decreased NREM sleep duration. REM sleep was not affected. Astressin (CRF receptor antagonist: i.p.) significantly attenuated the changes induced by urocortin, although astressin itself did not affect the sleep-wake pattern in the light period at this dosage. These findings show that urocortin changes the sleep-wake pattern in the light period. Moreover, urocortin was found to change the sleep-wake pattern by acting on CRF receptor, as astressin significantly attenuated the urocortin-induced changes on sleep-wake patterns. PMID- 17917259 TI - Protective mechanism of glycyrrhizin on acute liver injury induced by carbon tetrachloride in mice. AB - Glycyrrhizin is the major active component extracted from licorice (Glycyrrhiza glabra) roots, one of the most widely used herbal preparations for the treatment of liver disorders. This study evaluated the potential beneficial effect of glycyrrhizin in a mouse model of carbon tetrachloride (CCl(4))-induced liver injury. The mice were treated intraperitoneally with CCl(4) (0.5 ml/kg). They received glycyrrhizin (50, 100, 200, 400 mg/kg) 24 h and 0.5 h before and 4 h after administering CCl(4). The serum activities of aminotransferase and the hepatic level of malondialdehyde were significantly higher 24 h after the CCl(4) treatment, while the concentration of reduced glutathione was lower. These changes were attenuated by glycyrrhizin. CCl(4) increased the level of circulating tumor necrosis factor-alpha markedly, which was reduced by glycyrrhizin. The levels of hepatic inducible nitric oxide synthase, cyclooxygenase-2, and heme oxygenase-1 protein expression were markedly higher after the CCl(4) treatment. Glycyrrhizin diminished these alterations for inducible nitric oxide and cyclooxygenase-2 but the protein expression of heme oxygenase-1 was further elevated by the treatment of glycyrrhizin. CCl(4) increased the level of tumor necrosis factor-alpha, inducible nitric oxide synthase, cyclooxygenase-2, and heme oxygenase-1 mRNA expressions. The mRNA expression of heme oxygenase-1 was augmented by the glycyrrhizin treatment, while glycyrrhizin attenuated the increase in tumor necrosis factor-alpha, inducible nitric oxide synthase, and cyclooxygenase-2 mRNA expressions. These results suggest that glycyrrhizin alleviates CCl(4)-induced liver injury, and this protection is likely due to the induction of heme oxygenase-1 and the downregulation of proinflammatory mediators. PMID- 17917260 TI - Sex difference in ischemic acute renal failure in rats: approach by proteomic analysis. AB - It is known that female rats are resistant to ischemic acute renal failure (ARF), compared with male rats. To elucidate sex differences in ischemic ARF, we searched global protein expression in post-ischemic kidneys using proteomic techniques. Ischemic ARF was induced by 45-min ischemia followed by reperfusion. By proteomic analysis, many male- or female-dominant proteins were detected in sham-operated rat kidneys, and significantly increased or decreased proteins were found in post-ischemic kidneys 2 h after reperfusion, at which there were no significant deterioration in renal function of both sexes. We detected 86 proteins showing more than 1.5-fold significant alterations (p<0.01) in both sexes by ischemia/reperfusion (I/R) treatment. Among the altered proteins, we identified a significantly up-regulated protein in male rat kidneys, meprin alpha, a subunit of meprin which had been reported to play a role in the pathophysiology of I/R-induced ARF. In addition, it is known that a potent meprin alpha inhibitor, actinonin, can protect against I/R-induced renal injury when administered to male rats. We therefore compared the effect of actinonin on I/R induced renal dysfunction between male and female rats. Renal function of both males and females showed significant deterioration when measured at 24 h after the reperfusion, although the degree of renal dysfunction was much less in females than in males. Pre-ischemic treatment with actinonin (30 mg/kg, i.v.) prevented the I/R-induced renal dysfunction in males but not in females. Our results provide information on differences in protein expression at an early phase after the reperfusion between male and female rats. Moreover, the present study suggests that up-regulation of meprin alpha in the post-ischemic kidney is at least partly involved in aggravation of I/R-induced renal injury in male rats. The possibility that meprin alpha is a key component of the sex difference in ischemic ARF, warrants further attention. PMID- 17917261 TI - The glycosylation and pharmacokinetics of CTLA4Ig produced in rice cells. AB - Cytotoxic T-lymphocyte antigen 4-immunoglobulin (CTLA4Ig) has immunosuppressive activity and the ability to induce immune tolerance. There has been no report of its glycosylation ratio or of the role of its glycans. We investigated the terminal sialylation of rice cell-derived recombinant human CTLA4Ig (rrhCTLA4Ig) using lectins. The glycosylation ratios of rrhCTLA4Ig and Chinese hamster ovary (CHO) cell-derived recombinant human CTLA4Ig (crhCTLA4Ig) were evaluated by chemical deglycosylation. After intravenous (i.v.) or subcutaneous (s.c.) administration to rats, the pharmacokinetics of rrhCTLA4Ig and crhCTLA4Ig as well as of their deglycosylated forms were evaluated. rrhCTLA4Ig does not have terminal sialic acids and its glycosylation ratio was slightly lower than that of crhCTLA4Ig. Its terminal elimination half-life (T(1/2)) was shorter than that of crhCTLA4Ig following i.v. administration. However, the half-life was significantly prolonged and was similar with that of crhCTLA4Ig following s.c. administration. Moreover, the deglycosylated forms of both preparations were cleared from the circulation faster than the native forms. These results suggest that the presence of glycans on rrhCTLA4Ig and crhCTA4Ig are important for their in vivo stability. In addition, the glycan structure of rrhCTLA4Ig is more effective in maintaining in vivo stability after s.c. administration than after i.v. administration although the glycans on rrhCTLA4Ig lack terminal sialic acids, suggesting that its glycans have the potential for in vivo stability. PMID- 17917262 TI - In situ intestinal absorption behaviors of tanshinone IIA from its inclusion complex with hydroxypropyl-beta-cyclodextrin. AB - In this paper, the intestinal permeability of the inclusion complex of tanshinone IIA (TS IIA) with 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was investigated. The corresponding complexation of TS IIA-HP-beta-CD was obtained by coevaporation and characterized by differential scanning calorimetry and X-ray diffraction. The recirculation intestinal perfusion technique in rats was used to study the absorption behavior of free and complexed TS IIA. The change of concentration of TS IIA was separately calculated according to Michaelis-Menten and the Fick's equation to investigate its absorption rate-limiting step. Using the mathematical models above, it was concluded that the limit step to absorption of TS IIA was the dissolution process. Different concentrations of complexed TS IIA were administrated to three intestinal segments, with the intestinal permeability ranging from 3.16x10(-5) cm.s(-1) in the duodenum (50 microg.ml(-1)) to 4.11x10(-5) cm.s(-1) in the jejunum (100 microg.ml(-1)). With the increase of dosage of complex, TS IIA's absorption did not show saturated phenomenon, suggesting its transport mechanism in vivo might primary be passive transport. Besides, the permeability of TS IIA was not apparently influenced by the perfusion section studied, which indicated that there might not exist specific absorption site for TS IIA. PMID- 17917263 TI - Promising antitumor activity of a novel quinoline derivative, TAS-103, against fresh clinical specimens of eight types of tumors measured by flow cytometric DNA analysis. AB - TAS-103, 6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno-[2,1-c]quinolin-7 one dihydrochloride, is a dual topoisomerases I and II inhibitor. Antitumor activities of TAS-103 against fresh surgical specimens resected from 525 patients (32 types of tumors) were examined by flow cytometric (FCM) analysis of DNA integrity of tumor cells, and compared with those of five other investigational new drugs and 31 clinically available anticancer agents. Concentrations of clinically available anticancer agents were set at one-tenth of the peak plasma concentration (PPC) of the clinically recommended doses. On the other hand, since PPCs of investigational new drugs in humans were frequently unknown, these were estimated by a method that determines the theoretically achievable concentration in body fluid (TAC method). Correlations between TAC and PPC were examined for 16 clinically available anticancer agents, and it was found that TAC at 7n (the modified Fibonacci's dose-escalation scheme) of 14 drugs corresponded well with each one-tenth of PPC. By defining a 30% or more reduction in the integrated diploid peak as effective and a 60% or more reduction as definitely effective, TAS-103 at 5 microg/ml (7n) showed significantly higher effective rates and definitely effective rates than those of all other investigational new drugs, as well as almost all clinically available anticancer agents, against various malignancies, including non-small cell lung cancer, brain tumor and renal cancer. These results strongly suggest that TAS-103 will be expected to show excellent antitumor activities against a wide range of human tumors. PMID- 17917264 TI - Stereoselective metabolism of carvedilol by the beta-naphthoflavone-inducible enzyme in human intestinal epithelial Caco-2 cells. AB - Treatment of Caco-2 cells with beta-naphthoflavone (beta-NF) and 1alpha,25 dihydroxyvitamin D(3) (VD(3)) induces UDP-glucuronosyltransferases (UGTs) and cytochrome P450 (CYP) 3A4, respectively. In the present study, we evaluated the metabolism of carvedilol in beta-NF- and VD(3)-treated Caco-2 cells. The metabolism of R-carvedilol was not significant in non-treated Caco-2 cells, whereas S-carvedilol was significantly metabolized in the cells. The metabolism of R- and S-carvedilol was significantly increased by the treatment of Caco-2 cells with 50 microM beta-NF for 3 d. In contrast, the treatment of Caco-2 cells with 250 nM VD(3) for 2 weeks did not induce a significant change in the metabolism of R- and S-carvedilol. The metabolism of carvedilol in beta-NF treated Caco-2 cells was markedly inhibited by a substrate of UGTs, baicalein. In addition, the expression of UGT1A1, 1A6, and 1A9 mRNA was increased in beta-NF treated Caco-2 cells as compared with non-treated cells. These findings indicated that carvedilol was metabolized stereoselectively by the beta-NF-inducible enzyme in Caco-2 cells. The UGT1A subfamily in intestinal epithelial cells may be partly responsible for first-pass (presystemic) metabolism of the drug. PMID- 17917265 TI - Preventive effect of water containing magnesium ion on paw edema in adjuvant induced arthritis rat. AB - We demonstrate the preventive effect of bittern water (BW), which enables the effective intake of magnesium ion (Mg(2+)), on paw edema in adjuvant-induced arthritis (AA) rat. BW (five kinds; BW-1, 2, 3, 4, 5) containing 10-200 mg/l Mg(2+) was used in this study. Arthritis was induced by the injection of 50 microl of a suspension of 10 mg/ml heat-killed butyricum (Mycobacterium butyricum) in Bayol F oil into the plantar region of the right hind foot and tail of rats. Paw edema of the right and left hind feet of AA rats were reduced by the administration of BW for 14 d after adjuvant injection in comparison with those of AA rats administered purified water. The preventive effect increased with the increasing Mg(2+) content of the BW. In addition, a combination of indomethacin (IM, 2 mg/kg) and BW-5 (200 mg/l Mg(2+)) prevented paw edema of the right and left hind feet of AA rats in comparison with IM alone. The fate of plasma IM after the oral administration of the combined IM (2 mg/kg/d) and BW-5 was similar to that after the administration of IM alone. In conclusion, the oral administration of Mg(2+) to AA rats potently prevents the development of inflammation, and the combination of IM and Mg(2+) may provide an effective therapy of arthritic edema. PMID- 17917267 TI - Electrospray ionization and time-of-flight mass spectrometric method for simultaneous determination of spermidine and spermine. AB - A sensitive method for the determination of polyamines in mammalian cells was described using electrospray ionization and time-of-flight mass spectrometer. This method was 50-fold more sensitive than the previous method using ionspray ionization and quadrupole mass spectrometer. The method employed the partial purification and derivatization of polyamines, but allowed a measurement of multiple samples which contained picomol amounts of polyamines. Time required for data acquisition of one sample was approximately 2 min. The method was successfully applied for the determination of reduced spermidine and spermine contents in cultured cells under the inhibition of aminopropyltransferases. In addition, a new proper internal standard was proposed for the tracer experiment using (15)N-labeled polyamines. PMID- 17917266 TI - Constrictor responses to noradrenaline, hemodynamic profile, and superoxide levels measured by hydroethidine oxidation in diabetic rats. AB - The aim of this study was to test the hypothesis that vascular dysfunction in neonatal streptozotocin (n-STZ)-induced diabetic rats could be associated with alterations in blood pressure, hemodynamic profile, and levels of superoxide anion. Diabetes was induced by STZ injection (160 mg/kg, i.p.) in neonate (2-d old) Wistar rats. Using intravital microscopy the changes in mesenteric arteriolar diameters to vasoconstrictor agent noradrenaline (NA) and the levels of superoxide anion, measured by hydroethidine microfluorography, were determined in anaesthetized control and n-STZ rats. Blood pressure (BP) was determined in anaesthetized and unanaesthetized animals. Heart rate, shear rate, and blood flow velocity were also assessed. n-STZ rats showed, after 8 weeks of STZ injection, increased BP (unanaesthetized animals), hyperactivity to NA, and increased superoxide anion levels. However, heart rate, arteriolar shear rate, and blood flow velocity were unchanged in n-STZ. In conclusion, the results of the current study describe a significant increase in blood pressure, hyperactivity to NA mediated vasoconstriction, and increased superoxide levels measured by hydroethidine oxidation. Taken together, our findings demonstrate that the compromised ability of mesenteric microvessels to respond properly in n-STZ diabetic rats is associated with several vascular alterations. PMID- 17917268 TI - The validation of plasma darunavir concentrations determined by the HPLC method for protease inhibitors. AB - Darunavir (DRV) is a new protease inhibitor used to treat human immunodeficiency virus (HIV) type-1. The aim of this study was to validate the determination of plasma DRV concentrations using the HPLC method, a simple procedure for simultaneous determination of seven HIV protease inhibitors and efavirenz. The calibration curve was linear (range of 0.13 to 10.36 microg/ml). The average accuracy ranged from 100.7 to 105.6%. Both the interday and intraday coefficients of variation were less than 6.7%, which was similar to or much lower than previously reported values by the LC/MS/MS method. It is concluded that HPLC can be used to determine plasma DRV concentrations and routinely in the clinical setting; thus, this HPLC method enables further study of DRV pharmacokinetics in conventional hospital laboratories. PMID- 17917269 TI - Caspase-3-like protease activity-independent apoptosis at the onset of neuronal cell death in the gerbil hippocampus after global ischemia. AB - To investigate the relationship between caspase-3-like protease activity, which has been suggested to be related to apoptosis, and DNA fragmentation, we measured changes in caspase-3-like activity and DNA fragmentation in the hippocampus of gerbils exposed to global ischemia induced by bilateral occlusion of the carotid arteries for 5 min. Caspase-3-like protease activity began to increase at day 4 post-ischemia, reached a peak at day 5, and declined thereafter. The levels of DNA fragmentation, evaluated in terms of terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL) staining and cytosolic nucleosomes, in the ischemic hippocampus began to increase significantly at day 3 after ischemia, reached a peak at day 4, and decreased thereafter. Our data suggest that DNA fragmentation in ischemic hippocampus of gerbils precedes caspase-3-like protease activation. Our results indicate that a caspase-3-like protease-independent apoptotic pathway operates, at least at the onset of neuronal cell death, in the hippocampus of gerbils after global ischemia. PMID- 17917270 TI - Changes in the expression of cytochrome P450 genes in hemin-induced differentiated K562 cells. AB - We have previously reported the expression of CYP genes in human myeloblastic and lymphoid cell lines, and the induction of the CYP3A4 and GSTP1 genes by oxidative stress in the human erythroleukemia cell line, K562. To further elucidate the role of drug metabolizing enzymes in hematogenesis, we have characterized the expression of CYP genes in hemin-induced differentiated K562 cells. After incubation with 50 microM hemin for 3 d, the expression of CYP1A1 and CYP3A4 genes was induced by 2.5- and 3.5-fold, respectively. In contrast, the CYP1B1 and CYP2E1 genes were downregulated in these cells to below 10% of the control levels. Moreover, these changes correlated with the hemin dose and culture time. Metabolism of midazolam, a probe substrate for CYP3A4, in the differentiated K562 cells increased by 2-folds, suggesting that the induction of CYP3A4 activity is consistent with the mRNA level. If these changes in the CYP expression profile in hematopoietic cells occurred, the susceptibility to xenobiotics and/or the therapeutic drugs of the cells might be influenced, and it also affects the metabolism of endogenous substrates, such as steroids and prostaglandins. PMID- 17917271 TI - Increased chemotherapeutic activity of camptothecin in cancer cells by siRNA induced silencing of WRN helicase. AB - Werner syndrome helicase (WRN) participates in a wide range of DNA activities, including replication, double-strand DNA break repair, telomere and retrovirus long terminal repeat maintenance. Mutations of the WRN gene cause Werner syndrome (WS), an autosomal recessive premature ageing disorder associated with various symptoms related to ageing. In this study, we investigated the siRNA that specifically down-regulates WRN expression. WRN silencing increased markedly the chemotherapeutic activity of camptothecin (CPT) on cancer cells in terms of the extent of efficacy and lowering effective drug dosage, accompanied by suppressing recovery from DNA damage caused by CPT. Here, we propose a potential combination therapy of WRN-siRNA and CPT, looking forward to minimizing the inevitable adverse effects associated with cancer chemotherapy. PMID- 17917272 TI - Gene cloning and characterization of KdeA, a multidrug efflux pump from Klebsiella pneumoniae. AB - We cloned a gene related to multidrug resistance from a drug-resistant Klebsiella pneumoniae strain MGH78578. We designated the gene kdeA, which encodes a protein possessing 12 hydrophobic regions. The deduced amino acid sequence of KdeA is similar to that of MdfA, a well-characterized multidrug efflux pump from Escherichia coli. Introduction of the kdeA gene into cells of the drug hypersusceptible E. coli strain KAM32 resulted in elevated minimum inhibitory concentrations of chloramphenicol, norfloxacin, acriflavine, and ethidium bromide. We observed elevated energy-dependent ethidium efflux activity with cells carrying kdeA compared with control cells. We also observed expression of kdeA in cells of K. pneumoniae under normal growth conditions. PMID- 17917273 TI - Kakkalide and irisolidone: HMG-CoA reductase inhibitors isolated from the flower of Pueraria thunbergiana. AB - As part of our search for anti-arteriosclerosis agents from traditional Chinese medicines, the 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase (HCR)-inhibitory constituent, kakkalide, was isolated from the flower of Pueraria thunbergiana (PT, family Leguminosae). The antihyperlipidemic effects of kakkalide and its metabolite, irisolidone, which may be a bioactive form in vivo and potently inhibit the HCR activity, were investigated in vivo. Both the oral and interperitoneal administrations of kakkalide and irisolidone, with the exception of intraperitoneally treated kakkalide, potently lowered the serum levels of total cholesterol (TC) and triglyceride (TG) in Trition WR1339-induced hyperlipidemic mice. The oral administrations of kakkalide and irisolidone in hyperlipidemic mice induced, by the long-term feeding of a high fat diet, also potently reduced the serum levels of TC and TG and epididymal fat pad weight. These findings suggest that PT can improve hyperlipidemia, and the hypolipidemic effect may be due to HMG-CoA reductase. PMID- 17917274 TI - S-phase accumulation of Candida albicans by anticandidal effect of amentoflavone isolated from Selaginella tamariscina. AB - Amentoflavone, which is a biflavone derived from various plants, has been known to possess various biological activities including anti-tumor and anti inflammatory activity. In the previous study, we reported antifungal effect of amentoflavone isolated from an ethyl acetate extract of Selaginella tamariscina on human pathogenic fungi. Amentoflavone significantly inhibited the growth of fungal cells without hemolysis of human erythrocytes. In the present study, we investigated antifungal acting mode of amentoflavone in human pathogenic yeast Candida albicans. Anticandidal activity was exerted in an energy-independent manner presented by an antifungal assay in the presence of NaN(3), which is ATP depleting agent as a metabolic inhibitor. To investigate the effects of amentoflavone on cellular physiology in C. albicans, we performed cell cycle analysis, and the analysis showed that amentoflavone significantly arrested cell cycles during the S-phase. These results demonstrated that amentoflavone has potent anticandidal activity with significant physiological changes inducing S phase arrest in intracellular environment. Therefore, amentoflavone may be applied to a lead compound for the development of therapeutic agents, which can treat candidiasis resulted from candidal infections. PMID- 17917276 TI - The effects of glycine and L-arginine on heat stability of ginsenoside Rb1. AB - To identify the effects of amino acids on the heat stability of ginsenoside Rb(1) (Rb(1)), Rb(1) was heat-processed at 120 degrees C with or without glycine or L arginine. Rb(1) was changed into 20(S)-Rg(3), 20(R)-Rg(3), Rk(1), and Rg(5) by heat-processing through glycosyl elimination and epimerization of carbon-20 by SN1 reaction. Similarly, Rb(1) was changed into 20(S)-Rg(3), 20(R)-Rg(3), Rk(1), and Rg(5) when it was heat-processed with the same amount of glycine, but the generated amount of 20(S)-Rg(3) was higher than when Rb(1) was heat-processed without amino acids, and a significant increase in Maillard reaction products (MRPs) was noted. On the other hand, there were no structural changes in Rb(1) and the generation of MRPs when Rb(1) was heat-processed with the same amount of L-arginine. The improved heat stability of Rb(1) brought about by the addition of L-arginine was thought to be closely related to its characteristics of interfering with nonenzymatic glycation and forming hydrogen bonds with Rb(1). PMID- 17917275 TI - N-trans-feruloyltyramine as a melanin biosynthesis inhibitor. AB - In this study, we examined the effect of N-trans-feruloyltyramine (FA) on melanogenesis in mouse B16 melanoma cells. Melanogenesis was inhibited by FA in a dose-dependent manner. FA exhibited a greater potency than kojic acid as a standard inhibitor of melanogenesis. Moreover, treatment of B16 melanoma cells with FA was found to cause marked decreases in the expression levels of tyrosinase. FA-induced downregulation of tyrosinase resulted in suppression of melanin biosynthesis in murine B16 melanoma cells. PMID- 17917277 TI - Inhibitory effects of ginsenosides and their hydrolyzed metabolites on daunorubicin transport in KB-C2 cells. AB - We studied the effects of ginsenosides and their metabolites on daunorubicin transport in multidrug-resistant P-glycoprotein (P-gp)-overexpressing KB-C2 cells. Ginsenoside Rg1, which is a protopanaxatriol-type ginseng saponin, did not have any effects on the accumulation of P-gp substrate daunorubicin. On the other hand, its metabolite M4, which has no sugar moiety, increased the accumulation 3.6-fold at 5 microM. Metabolites of protoanaxadiol-type saponin M1 and M12 also increased accumulation, but the effects were less than that of M4. The findings showed larger effects of metabolites without glucose moieties. Analysis of verapamil-stimulated ATPase activity in membrane vesicles expressing human P-gp suggested that the increased daunorubicin accumulation by M4 was at least partly due to ATPase inhibition of P-gp. PMID- 17917278 TI - Effects of capsaicin on cellular damage and monolayer permeability in human intestinal Caco-2 cells. AB - Recent studies suggest that capsaicin (Cap), a major constituent of hot pepper, may affect the function and permeability of the intestinal mucosa in vitro. However, the relationships between the dose of Cap and the barrier and/or transporter functions on intestinal epithelial cells are unknown. The aim of this study was to investigate whether Cap initiates cellular injury and alter epithelial permeability in Caco-2 cells. Cellular toxicity, as measured using a lactate dehydrogenase release assay, was not observed at high concentrations of Cap (up to 300 microM). When cell viability was measured by a WST-1 assay (tetrazolium salt-based assay), damage to Caco-2 monolayers was observed at doses of 200 and 300 microM of Cap. The barrier function of tight junctions was assessed by measuring transepithelial electrical resistance (TEER) in Caco-2 cells. Treatment of Caco-2 cells with Cap at doses above 100 microM significantly decreased the TEER compared to treatment with buffer alone for 2 h (p<0.05). We next examined the effects of Cap on the activity of P-glycoprotein (P-gp) found on transcellular transporters. At doses of 100 and 200 microM, Cap inhibited the transport of rhodamine 123 by P-gp-mediated efflux in Caco-2 cells. Cap thus exhibited inhibitory effects on P-gp. The results of this study indicate that Cap, a dietary phytochemical, causes functional and structural changes in Caco-2 cell monolayers at noncytotoxic doses (less than 100 microM of Cap). The concomitant administration of Cap with drugs that are substrates of P-gp might increase the plasma concentrations of such drugs. PMID- 17917279 TI - Kinetic analysis of effects of mouth washing on removal of drug residues following inhalation of fluticasone propionate dry powder. AB - Fluticasone propionate dry powder inhaler (FP-DPI) is widely used for the treatment of asthma. However, local adverse effects such as oropharyngeal candidiasis are often seen and mouth washing after inhaling is recommended. In our previous study, we reported a nonlinear relationship between the amount of drug residue and number of times mouth washing was employed. Thus, we developed a compartment model, in which the inhaled drugs were distributed in both easy and difficult to remove areas. Using this model, we analyzed drug removal efficiency in each area with different mouth washing procedures. Three methods of mouth washing were studied; gargling and rinsing in combination, rinsing alone, and gargling alone, following administration of FP-DPI by sprinkling or inhaling. The amounts of drugs recovered from areas considered to be easy to remove (X(1)) and difficult to remove (X(2)) were determined using a nonlinear least-squares program, while the removal efficiency of each of the 3 methods was also calculated. The ratios of X(1) after sprinkling and inhalation were 63.9% and 21.8%, respectively, while those of X(2) were 6.0% and 12.4%, respectively. The numbers of mouth washings required to remove half doses from easy and difficult to remove areas were 0.2 and 1.4 times, respectively, with a combination of gargling and rinsing following inhalation of FP-DPI, while those were 0.3 and 3.6 times, respectively, with rinsing alone, and 0.4 and 5.8 times, respectively, with gargling only, thus demonstrating significant differences among the mouth washing methods for efficiency in the difficult to remove area. The present results show that the employed methods of mouth washing had a significant influence on the removal of drug residues following inhalation of FP-DPI, with gargling and rinsing in combination considered to be the most effective. PMID- 17917280 TI - Identification of a tryptanthrin metabolite in rat liver microsomes by liquid chromatography/electrospray ionization-tandem mass spectrometry. AB - Tryptanthrin originally isolated from Isatis tinctoria L. has been characterized to have anti-inflammatory activities through the dual inhibition of cyclooxygenase-2 and 5-lipoxygenase mediated prostaglandin and leukotriene syntheses. To characterize phase I metabolite(s), tryptanthrin was incubated with rat liver microsomes in the presence of NADPH-generating system. One metabolite was identified by liquid chromatography/electrospray ionization-tandem mass spectrometry. M1 could be identified as a metabolite mono-hydroxylated on the aromatic ring of indole moiety from the MS(2) spectra of protonated tryptanthrin and M1. The structure of metabolite was confirmed as 8-hydroxytryptanthrin with a chemically synthesized authentic standard. The formation of M1 was NADPH dependent and was inhibited by SKF-525A, a general CYP-inhibitor, indicating the cytochrome P450 (CYP)-mediated reaction. In addition, it was proposed that M1 might be formed by CYP 1A in rat liver microsomes from the experiments with enriched rat liver microsomes. PMID- 17917281 TI - No association of the G1287A polymorphism in the norepinephrine transporter gene and susceptibility to major depressive disorder in a Japanese population. AB - Norepinephrinergic neurotransmission in the central nervous system have a major impact on the symptomatology in major depressive disorder (MDD), and genetic polymorphisms of norepinephrine transporter (NET) have a possibility to be involved in susceptibility to MDD. We investigated the association of the G1287A (rs5569) polymorphism of the NET gene and susceptibility to MDD by comparing 145 major depressive patients with 164 healthy individuals first in a Japanese population. The genotype frequencies in depressed patients and health volunteers of the NET G1287A polymorphism were 52.4% (G/G), 39.3% (G/A) and 8.3% (A/A) in depressed patients, 61.6% (G/G), 29.9% (G/A allele) and 8.5% (A/A) in healthy volunteers, respectively. The allele frequencies in depressed patients and health volunteers of the NET G1287A polymorphism were 72.1% (G allele) and 27.9% (A allele) in depressed patients, 76.5% (G allele) and 23.5% (A allele) in healthy volunteers, respectively. The genotype distribution and allele frequencies were not significantly different between major depressive patients and healthy volunteers. NET G1287A polymorphism appears not to be an important factor in susceptibility to MDD in a Japanese population. PMID- 17917282 TI - Determination of diclofenac sodium in commercial pharmaceutical formulations and human control serum using a kinetic-spectrophotometric method. AB - A kinetic method for the determination of micro quantities of diclofenac sodium (DS) is described in this paper. The method is based on a ligand-exchange reaction. The reaction was followed spectrophotometrically by monitoring the rate of appearance of the cobalt diclofenac complex at 376 nm. The optimum operating conditions regarding reagent concentrations and temperature were established. The optimized conditions yielded a theoretical detection limit of 1.29 microg ml(-1) based on the 3S(b) criterion. The interference effects of certain drugs, foreign ions and amino acids upon the reaction rate were studied in order to assess the selectivity of the method. The developed procedure was successfully applied to the rapid determination of diclofenac sodium in commercial pharmaceutical preparations and human control serum. The unique features of this procedure are that determination can be carried out at room temperature and the analysis time is short. The newly developed method is simple, inexpensive, and efficient for use in the analysis of a large number of samples. PMID- 17917283 TI - Optimization of an HPLC method for determination of gabapentin in dosage forms through derivatization with 1-fluoro-2,4-dinitrobenzene. AB - A rapid, sensitive and accurate high performance liquid chromatography with UV detection method was developed and validated for the quantification of gabapentin in dosage forms. Gabapentin was quantified after pre-column derivatization with 1 fluoro-2,4-dinitrobenzene. Amlodipine was used as an internal standard. The chromatographic separation was carried out on a Nova-Pak C(18) column using a mixture of acetonitrile-sodium dihydrogenphosphate (pH 2.5; 0.05 M) (70:30, v/v) as mobile phase with UV detection at 360 nm. The method was linear over the range of 10-500 microg/ml of gabapentin (r(2)>0.999). The within-day and between-day precision values were in the range of 0.86-1.11%. The method was successfully used for quantitative determination and dissolution rate study of Neurontin capsules. PMID- 17917284 TI - Study of compounds suppressing free radical generation from UV-exposed ketoprofen. AB - Ketoprofen [(RS)-2-(3-benzoylphenyl)propanoic acid] is widely used for the treatment of inflammatory diseases and musculoskeletal injury. However, there is concern regarding its potential for photosensitization as a side effect. Free radicals and active oxygen species generated from ketoprofen on exposure to ultraviolet (UV) light have been implicated in phototoxicity and photosensitization. In this study, we examined the suppressing ability of some compounds for the free radicals and active oxygen species generated by the photodynamic reaction of ketoprofen, to determine a new resist of photosensitization by ketoprofen. Eight compounds, including six known free radical scavengers were individually mixed with ketoprofen, and the mixtures were exposed to UV. Then, the free radicals and the active oxygen species were determined by the electron spin resonance spectrometry to estimate suppressing and scavenging ability of compounds. The compounds that show promise in suppressing superoxide anion generation from UV-exposed ketoprofen were further evaluated using the on-line photo-irradiated superoxide anion detection system. It was confirmed that quercetin, a flavonoid, strongly suppresses the generation of free radicals and active oxygen species from UV-exposed ketoprofen. The experiments using the experimental formulation of an adhesive skin patch of ketoprofen containing quercetine and the Chemiluminescence analyzer (CLA) indicated that quercetin has high potential for use as an excipient in ketoprofen ointments to suppress phototoxicity and photosensitization by ketoprofen. PMID- 17917285 TI - Selective synthesis and utility of one tripyrrolic compound and its intermediates. AB - Highly selective syntheses of tri(2,4-dimethyl-3-carbethoxypyrrolyl)-methane 8 and its dipyrrolic intermediate 6 and pyrrolic one 1 are described based on the successful correction of the wrong process for 1 in literature. Tripyrrolic compounds have attracted much attention recently and been developed in diverse fields. 1 was the key intermediate for some tyrosine kinase inhibitors, including newly-launched Sutent, and most recently we have found 6 was also synthetically useful in the synthesis of 11 that has been discovered as a novel histone deacetylases (HDAC) inhibitor with an IC(50) value of about 1 microM in our assessments and represents a promising lead for the development of more potent histone deacetylase inhibitors (HDACIs). PMID- 17917286 TI - Apotirucallane and tirucallane triterpenoids from Cedrela sinensis. AB - Nine new triterpenoids, 1-9, were isolated from the cortex of Cedrela sinensis (Meliaceae), together with six known compounds, sapelin E acetate, grandifoliolenone, azadirone, bourjotinolone A, piscidinol A, and hispidol B. The structures of 1-9 were determined by the 2D NMR experiments, chemical methods, and X-ray crystallography. PMID- 17917287 TI - Application of polyglycolized glycerides in protection of amorphous form of etoricoxib during compression. AB - Polymorphic transition and stability problems during amorphous drug formulation are the major limiting factors in pharmaceutical technology. The purpose of the study was to evaluate the ability of polyglycolized glycerides (Gelucire) in protection of amorphous form of drug during compression and shelf life with lower proportion. Amorphous etoricoxib (AET) was prepared by spray drying technique. Tablets of AET and melt granules of AET (MG-AET) with Gelucire 50/13 were prepared. Tablets parameters like hardness, disintegration and content uniformity were evaluated. Tablets were evaluated immediately after compression and on storage for 3 months at ambient conditions to determine degree of transformation using X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC) and dissolution profiles. Spray drying yielded the amorphous etoricoxib. Content uniformity in the tablet was in between 95 to 105%. Other parameters like disintegration and hardness were well within the limits. The results showed significant difference in the degree of crystallinity between AET tablet and MG AET tablet. MG-AET tablet showed absence of crystallinity after 3 months storage. The reason could be formation of hydrogen bonding between the Gelucire and AET. Also Gelucire can be tableted very easily under low pressure and showed elastic recovery. Gelucire yielded a soft embedding during tableting, which prevented the polymorphic transformation. Polyglycolized glycerides (Gelucire 50/13) are able to protect amorphous etoricoxib during compression. As excipient required is low, it became possible to prepare tablet formulation as compared to other excipient like polyvinylpyrrolidon (PVP). PMID- 17917288 TI - The suppression of enhanced bitterness intensity of macrolide dry syrup mixed with an acidic powder. AB - The aim of the present study was to identify a medicine which strongly enhanced the bitterness of clarithromycin dry syrup (CAMD) when administered concomitantly and to develop a method to suppress this enhanced bitterness. The bitterness enhancement was evaluated not only by gustatory sensation tests but also using pH and taste sensor measurements of the mixed sample. A remarkable bitterness enhancement was found when CAMD was mixed with the acidic powder L-carbocysteine. The acidic pH (pH 3.40) of the suspension made from these two preparations, seemed to be due to enhanced release of clarithromycin caused by the dissolution of the alkaline polymer film-coating. Several methods for preventing this bitterness enhancement were investigated. Neither increasing the volume of water taken with the mixture, nor changing the ratio of CAMD:L-carbocysteine in the mixture, were effective in reducing the bitterness intensity of the CAMD/L carbocysteine mixture. The best way to achieve taste masking was to first administer CAMD mixed with chocolate jelly, which has a neutral pH, followed by the L-carbocysteine suspension. Similar results were obtained for the bitterness suppression of azithromycin fine granules with L-carbocysteine. The chocolate jelly will be useful for taste masking of bitter macrolide drug formulations, when they need to be administered together with acidic drug formulations. PMID- 17917289 TI - Preparation of new nitrogen-bridged heterocycles. 60. Syntheses and conformational analyses of bis(indolizin-1-yl) disulfides. AB - Some bis(indolizin-1-yl) disulfides, readily obtainable from the treatment of 1 (benzoylthio)indolizines with piperidine, were prepared and their conformations were investigated. In comparison with those of 1-(benzoylthio)indolizines, the (1)H-NMR spectra of these disulfides showed considerable high field shifts (delta 0.13-0.82 ppm) on each pyridine ring proton and the UV spectra exhibited significant bathochromic and hyperchromic shifts. These results supported strongly the participation of an intramolecular pi-pi interaction between the two indolizine rings in these molecules and, hence, of a particular gauche (cis) conformation. However, the conformational considerations and molecular calculations (Mopac PM3) for some bis(indolizin-1-yl) disulfides showed the presence of four more stable gauche forms in which two are enantiomeric, resulting in three types of gauche structures. These three types of gauche structures were confirmed by X-ray analyses. PMID- 17917290 TI - Improvement of dissolution properties of a new Helicobacter pylori eradicating agent (TG44) by inclusion complexation with beta-cyclodextrin. AB - The interaction of a newly developed Helicobacter pylori eradicating agent (TG44, 4-methylbenzyl-4'-[trans-4-(guanidinomethyl)cyclohexylcarbonyloxy]biphenyl-4 carboxlylate monohydrochloride) with beta-cyclodextrin (beta-CyD) in aqueous solution and in solid state was studied to gain insight into the high in-vivo H. pylori eradicating activity of TG44/beta-CyD complex. The interaction was studied by the solubility method, spectroscopic methods, powder X-ray diffractometry and differential scanning colorimetry (DSC). TG44 gave A(L)-type phase solubility diagram with beta-CyD in water, showing a linear increase in solubility of the drug up to 8 mM beta-CyD concentration. The solubility of TG44 (0.04 mM in water at 25 degrees C) increased about 70-folds at 8 mM beta-CyD. Ultraviolet, circular dichroism, fluorescence and (1)H-nuclear magnetic resonance spectroscopic studies indicated that TG44 forms the inclusion complex with beta-CyD in a 1:1 stoichiometry and the biphenyl moiety of TG44 is preferably included in the beta CyD cavity in water. The Giordano plot made by monitoring changes in the fusion enthalpy of TG44 (about 184 degrees C) suggested that TG44 forms the 1:1 complex with beta-CyD in the solid state. The TG44/beta-CyD solid complex in a 1:1 stoichiometry was prepared by the grinding and spray-drying methods and confirmed by powder X-ray diffractometry and DSC that the complex is in an amorphous state. The initial dissolution rate of TG44/beta-CyD complex was significantly faster than those of the drug alone and the physical mixture of both components, maintaining higher supersaturated concentrations of the drug for a long time. The results suggested that the higher eradicating activity of TG44/beta-CyD complex to Helicobacter pylori, compared with that of the drug alone, is attributable at least partly to the faster dissolving property of the complex and its ability to maintain the supersaturated state of the drug in the gastric fluid. PMID- 17917291 TI - Xeniaphyllane-derived terpenoids from the formosan soft coral Sinularia gibberosa. AB - New xeniaphyllane-derived metabolites (1-7) were isolated from the EtOAc extract of the Formosan soft coral Sinularia gibberosa. The structures and relative configurations of these compounds were elucidated on the basis of extensive spectroscopic analysis (including 2D NMR) and by comparison of their spectral data with those of related compounds. In vitro cytotoxic evaluation of the above metabolites towards a limited panel of cancer cell lines is also described. PMID- 17917292 TI - Estrogenic and anti-estrogenic activities of Cassia tora phenolic constituents. AB - Through an estrogenic activity bioassay-guided fractionation of the 70% ethanolic extract of Cassia tora seeds two new phenolic triglucosides, torachrysone 8-O [beta-D-glucopyranosyl(1-->3)-O-beta-D-glucopyranosyl(1-->6)-O-beta-D glucopyranoside] (1) and toralactone 9-O-[beta-D-glucopyranosyl-(1-->3)-O-beta-D glucopyranosyl-(1-->6)-O-beta-D-glucopyranoside] (2), along with seven known compounds were isolated. The structures of the new compounds were elucidated on the basis of spectroscopic and chemical evidence. The estrogenic activity of the fractions and the isolated compounds were investigated using the estrogen dependent proliferation of MCF-7 cells. In addition, the yeast two hybrid assay expressing estrogen receptor alpha (ERalpha) and beta (ERbeta) and the ERalpha competitor screening assay (ligand binding screen) were used to verify the binding affinities of the isolated compounds to ER. Furthermore, a naringinase pre-treatment of the 70% alcoholic extract of Cassia tora seeds resulted in a significant increase in its estrogenic activity. From the naringinase pre-treated extract six compounds were isolated, among which 6-hydroxymusizin and aurantio obtusin showed the most potent estrogenic activity, while torachrysone, rubrofusarin and toralactone showed a significant anti-estrogenic activity. Finally, the structure requirements responsible for the estrogenic activity of the isolated compounds were studied by investigating the activity of several synthetic compounds and chemically modifying the isolated compounds. The basic nucleus 1,3,8-trihyroxynaphthalene (T(3)HN) was found to play a principal role in the binding affinity of these compounds to ER. PMID- 17917293 TI - Electroreduction of the muscle relaxant drug dantrolene sodium at the mercury electrode and its determination in bulk form and pharmaceutical formulation. AB - The electroreduction of the muscle relaxant drug dantrolene sodium at the mercury electrode has been studied in the Britton-Robinson universal buffer of pH 2.5 11.5 containing 20% (v/v) methanol by means of dc-polarography, cyclic voltammetry and controlled-potential coulometry. Its reduction took place via three irreversible cathodic steps in solutions of pH < or =6, two steps in solutions of 6 or =10 through the consumption of 10, 8 or 4 electrons, respectively. This behavior was attributed to the reduction of NO(2) group (1st and 2nd steps at pH < or =6 or the single step at pH > or =10) and the -CH=N- double bond (3rd step at pH <10). Two polarographic procedures (direct current and differential-pulse modes) and three adsorptive cathodic stripping voltammetric procedures (linear-sweep, differential pulse and square-wave modes) were described and successfully applied for quantification of dantrolene sodium in its bulk form and in pharmaceutical formulation (Dantrolex tablets). PMID- 17917294 TI - Catalytic aerobic oxidation of nor-binaltorphimine (nor-BNI) analogs without 4,5 epoxy bridge affords a more selective ligand for kappa opioid receptor than the representative kappa antagonist nor-BNI. AB - An analog of nor-binaltorphimine (nor-BNI) without the 4,5-epoxy bridge, 17,17' bis(cyclopropylmethyl)-6,6',7,7'-tetrahydro-6,6'-imino-14beta,14'alpha-dihydroxy 3,3'-dimethoxy-7,7'-bimorphinan (4), which was the precursor of the designed compound 1 as a selective kappa(3) opioid receptor antagonist, was catalytically oxidized with oxygen in the presence of platinum to give the 5'-oxo derivative 3 with some other oxidized products. Morphinan derivatives without the 4,5-epoxy moiety were labile to oxygen, although the corresponding 4,5-epoxymorphinan derivatives resisted aerobic oxidation. One of the oxidized nor-BNI analogs without 4,5-epoxy bridge, compound 18, showed high affinity and selectivity for kappa opioid receptor. PMID- 17917295 TI - Cinnamylindoline derivatives: synthesis and factor Xa (FXa) inhibitory activities. AB - A series of cinnamylindoline derivatives were synthesized, and their factor Xa (FXa) inhibitory activities and selectivity over trypsin were evaluated. Among them, some novel derivatives showed potent FXa inhibitory activities and good selectivity over trypsin. Especially, (E)-2-{5-[1-(acetimidoyl)piperidin-4-yloxy] 2-[2-(5-amidino-2-hydroxyphenyl)ethen-1-yl]indolin-1-ylsulfonyl}acetic acid (22f) having 2-hydroxycinnamyl moiety exhibited the most potent FXa inhibitory activity in vitro. Furthermore, 22f also exhibited potent anticoagulant activities in vitro. PMID- 17917297 TI - Microbial metabolism. Part 8. The pyranocoumarin, decursin. AB - Microbial transformation of the cancer chemopreventive agent, decursin (1) with Sepedonium chrysospermem (ATCC 13378) yielded two metabolites, (+)-decursinol (2) and (-)-cis-decursidinol (3). The structures were established by spectroscopic data. PMID- 17917296 TI - Bioactive constituents from Chinese natural medicines. XXVI. Chemical structures and hepatoprotective effects of constituents from roots of Rhodiola sachalinensis. AB - The methanolic extract from the roots of Rhodiola sachalinensis was found to show a protective effect on D-galactosamine-induced cytotoxicity in primary cultured mouse hepatocytes. From the methanolic extract, five new glycosides, two monoterpene glycosides, two flavonol bisdesmosides, and a cyanogenic glycoside, were isolated together with 34 known compounds. The structures of new constituents were elucidated on the basis of chemical and physicochemical evidence. In addition, the principal constituents, sachalosides III and IV, rhodiosin, and trans-caffeic acid, displayed hepatoprotective effects. PMID- 17917298 TI - A facile synthesis of p- and m-(amidinomethyl)phenyl esters derived from amino acid and tryptic hydrolysis of these synthetic inverse substrates. AB - A facile synthetic method for p- and m-(amidinomethyl)phenyl esters derived from a variety of amino acids is presented. We analyzed the kinetic behavior of trypsin towards these synthetic esters, which are inverse substrates. The substituent (meta- and para-isomers) and isosteric effects of (amidinomethyl)phenyl esters are discussed. PMID- 17917299 TI - Chemical phenotypes of the hmg1 and hmg2 mutants of Arabidopsis demonstrate the in-planta role of HMG-CoA reductase in triterpene biosynthesis. AB - Plants produce a wide variety of cyclic triterpenes, such as sterols and triterpenoids, which are the major products of the mevalonate (MVA) pathway. It is important to understand the physiological functions of HMG-CoA reductase (HMGR) because HMGR is the rate-limiting enzyme in the MVA pathway. We have previously isolated Arabidopsis mutants in HMG1 and HMG2. Although the biochemical function of HMGR2 has been thought to be almost equal to that of HMGR1, based on similarities in their sequences, the phenotypes of mutants in these genes are quite different. Whereas hmg2 shows no abnormal phenotype under normal growth conditions, hmg1 shows pleiotropic phenotypes, including dwarfing, early senescence, and male sterility. We previously postulated that the 50% decrease in the sterol content of hmg1, as compared to that in the wild type, was a cause of these phenotypes, but comprehensive triterpene profiles of these mutants had not yet been determined. Here, we present the triterpene profiles of hmg1 and hmg2. In contrast to hmg1, hmg2 showed a sterol content 15% lower than that of the wild type. A precise triterpenoid quantification using synthesized deuterated compounds of beta-amyrin (1), alpha-amyrin (2), and lupeol (3) showed that the levels of triterpenoids in hmg1 and hmg2 were 65% and 25% lower than in the wild type (WT), respectively. These results demonstrate that HMGR2 as well as HMGR1 is responsible for the biosynthesis of triterpenes in spite of the lack of visible phenotypes in hmg2. PMID- 17917300 TI - Marked production of ginsenosides Rd, F2, Rg3, and compound K by enzymatic method. AB - The hydrolysis of protopanaxadiol-type saponin mixture by various glycoside hydrolases was examined. Among these enzymes, crude preparations of lactase from Aspergillus oryzae, beta-galactosidase from A. oryzae, and cellulase from Trichoderma viride were found to produce ginsenoside F(2) [3-O-(beta-D glucopyranosyl)-20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol], compound K [20 O-beta-D-glucopyranosyl-20(S)-protopanaxadiol], and ginsenoside Rd {3-O-[beta-D glucopyranosyl-(1-->2)-beta-D-glucopyranosyl]-20-O-beta-D-glucopyranosyl-20(S) protopanaxadiol}, respectively, from protopanaxadiol-type saponin mixture in large quantities. Moreover, the crude preparation of lactase from Penicillium sp. having a high producing activity of ginsenoside Rh(1) (6-O-beta-D-glucopyranosyl 20(S)-protopanaxatriol) from protopanaxatriol-type saponin mixture gave ginsenoside Rd as a main product, ginsenoside Rg(3) {3-O-[beta-D-glucopyranosyl (1-->2)-beta-D-glucopyranosyl]-20(S)-protopanaxadiol}, and compound K from protopanaxadiol-type saponin mixture. The hydrolytic pathways of ginsenosides Rb(1), Rb(2), and Rc to ginsenosides Rd, Rg(3), and F(2), and compound K by crude preparations of four glycoside hydrolases were also studied. This is the first report on the enzymatic preparation of an intestinal bacterial metabolite, ginsenoside F(2), in quantity, and a considerable amount of a minor saponin, ginsenoside Rg(3), from a protopanaxadiol-type saponin mixture. PMID- 17917301 TI - Compositions of royal jelly II. Organic acid glycosides and sterols of the royal jelly of honeybees (Apis mellifera). AB - Two organic acid glycosides (1, 2) and 16 sterols were isolated from the royal jelly of honeybees (Apis mellifera). The former two were monoglucosides of 10 hydroxy-2E-decenoic and 10-hydroxydecanoic acids. They are the first examples of glycosides isolated from royal jelly. The latter 16 were sterols mainly composed of 28 or 29 carbons. Among them, four compounds were new isofucosterol derivatives, and their structures were characterized as (24Z)-stigmasta-5,24(28) dien-3beta-ol-7-one (3), (24Z)-stigmasta-5,24(28)-diene-3beta,7beta-diol (4), (24Z)-stigmasta-5,24(28)-diene-3beta,7alpha-diol (5), and (24Z)-stigmast-24(28) ene-3beta,5alpha,6beta-triol (6) on the basis of various NMR spectroscopic data. PMID- 17917302 TI - Clerodane diterpenoids and flavonoids with NGF-potentiating activity from the aerial parts of Baccharis gaudichaudiana. AB - A new clerodane diterpene, 15-hydroxy-16-acetoxy-ent-clerod-3-en-18-oic acid (1), together with three known clerodane diterpenes (2-4) and three known flavones (5 7), were isolated from the aerial parts of Baccharis gaudichaudiana. Their structures were elucidated on the basis of spectroscopic analysis. Compounds 2, 3, and 5 showed enhancing activity of nerve growth factor (NGF)-induced neurite outgrowth in PC 12D cells. PMID- 17917303 TI - Resveratrol tetramers with a C6-C3 or a C1 unit from Upuna borneensis. AB - Investigation of the chemical constituents in the stem of Upuna borneensis (Dipterocarpaceae) resulted in the isolation of three new resveratrol derivatives, upunaphenols L (1), M (2) (resveratrol tetramers with a C(6)-C(3) unit) and N (3) (resveratrol tetramer with a C(1) unit). The structures have the same partial structure as vaticanol B (4). Upunaphenols L and M are new complex polyphenol compounds, lignostilbene. Their structures were determined by spectroscopic analysis including two dimensional NMR. Upunaphenol M was found to be an artifact generated by silica gel catalyzed methanolysis of 1. PMID- 17917304 TI - Selective hydrogenation of alkene in (3-trifluoromethyl) phenyldiazirine photophor with Wilkinson's catalyst for photoaffinity labeling. AB - Selective hydrogenation of carbon-carbon double bond in the presence of nitrogen nitrogen double bond in (3-trifluoromethyl) phenyldiazirine achieved with Wilkinson's catalyst. PMID- 17917305 TI - A novel function of plant histone H1: microtubule nucleation and continuous plus end association. AB - In higher plant cells, various microtubular arrays can be seen despite of their lack of structurally defined microtubule-organizing centers (MTOCs) like centrosomes in animal cells. Little is known about the molecular properties of the microtubule-organizing centers in higher plant cells. The nuclear surface contains one of these microtubule-organizing centers and generates microtubules radially toward the cell periphery (radial microtubules). Previously, we reported that histone H1 possessed the microtubule-organizing activity, and it was suggested that histone H1 localized on the nuclear surfaces in Tobacco BY-2 cells (Nakayama, T., Ishii, T., Hotta, T., and Mizuno, K. J. Biol. Chem. (submitted)). Here we show that histone H1 forms ring-shaped complexes with tubulin, and these complexes nucleated and elongated the radial microtubules continuously (processively) associating with their proximal ends where the incorporation of tubulin occurred. Furthermore, the polarity of radial microtubules was determined to be proximal end plus. Immunofluorescence microscopy of the isolated nuclei revealed that histone H1 localized on the nuclear surfaces, distinct from that in the chromatin. These results indicate that radial microtubules are organized by a novel MTOC that is totally different from MTOCs previously found in either plant or animal cells. PMID- 17917306 TI - Successful intrauterine therapy for fetal goitrous hypothyroidism during late gestation. AB - We experienced a case of fetal goitrous hypothyroidism in an infant delivered by a 33-year-old woman receiving 300 mg/day of propylthiouracil (PTU) for hyperthyroidism due to Graves' disease. A large fetal goiter (maximum diameter, 60 mm) was detected by magnetic resonance imaging (MRI) at 36 weeks of gestation. Initial fetal blood sampling revealed hypothyroidism with a serum thyroid stimulating hormone (TSH) of 99 microIU/mL, free triiodothyronine (T(3)) of 1.97 pg/mL, and free thyroxine (T(4)) of 0.29 ng/dL. Consequently, a diagnosis of fetal goitrous hypothyroidism due to transplacental passage of maternal PTU was made. To reduce the risk of perinatal complications, 300 microg of levothyroxine sodium (L-T(4)) was administered into the maternal amniotic fluid twice between 37 and 38 weeks of gestation. Subsequent fetal MRI showed that the size of goiter had decreased. At 38 weeks and 5 days of gestation, a 3042-g male infant was born by cesarean section. There were no severe complications at delivery, although mild tachypnea was observed and the infant's thyroid gland was slightly enlarged. He was treated with L-T(4) for two weeks. At present, his growth and neurological development are normal. This case indicates that intrauterine therapy by the intraamniotic administration of L-T(4) can be effective in treating fetal goitrous hypothyroidism even during late gestation. PMID- 17917307 TI - Clinical significance of 1-year treatment with raloxifene on bone and lipid metabolism in Japanese postmenopausal women with osteoporosis. AB - It has been well established that raloxifene (RLX) has beneficial effects on bone primarily in Caucasian women. However, to date, there is a dearth of data for Japanese postmenopausal women. In this study, we prospectively evaluated the effects of RLX on bone and lipid metabolism in fifty Japanese postmenopausal patients with untreated osteoporosis. We measured bone mineral density (BMD) by dual-energy X-ray absorptiometry at 7 sites including the lumbar spine, femoral neck, and distal radius. BMD was significantly increased at the lumbar spine both at 6 months and at 12 months compared with at baseline (p<0.01 for both), although the possibility could not be completely excluded that this increase may be partly explained by an apparent increase induced by degenerative changes in lumbar vertebrae since we had no control subjects to compare and be more certain of the findings in this study. Both bone-specific alkaline phosphatase (BAP) and serum N-terminal telopeptide of type I collagen (NTx) significantly decreased both at 6 months (p<0.01 for both) and at 12 months (p<0.01 for both) compared with at baseline, but not below the lower limit of the reference value. Total cholesterol and low-density lipoprotein cholesterol were significantly improved while triglycerides and high-density lipoprotein cholesterol were unaltered. Although longer and larger studies with fracture endpoints are needed to draw definite conclusions, our findings suggest the favorable effects of RLX on bone and lipid metabolism in Japanese postmenopausal women with osteoporosis as in Caucasian women. PMID- 17917308 TI - Toxic thyroid adenoma presenting as hypokalemic periodic paralysis. AB - Toxic thyroid adenoma presenting as hypokalemic periodic paralysis is extraordinarily rare. We describe a 26-year-old Japanese man who suffered from acute and painful muscle weakness of extremity in the morning. Physical examination showed a left anterior neck mass and laboratory tests revealed hypokalemia during his paralysis, and thyrotoxicosis. Neck sonogram showed a solitary nodule in the left lobe of the thyroid. Thyroid scintigraphy revealed a hot nodule of the tumor region with suppressed uptake in the other thyroid area. The tumor was surgically removed and his paralytic attack ceased. No somatic mutation of TSH receptor was found in his thyroid adenoma and no known genetic mutations of ionic channel genes, such as calcium (CACN1S), sodium (SCN4A) and potassium (KCNE3), were found. Although thyrotoxic periodic paralysis is usually accompanied with Graves' disease, thyrotoxicosis of other conditions including Plummer's disease should be considered. PMID- 17917309 TI - Defective expression of prohormone convertase 1/3 in silent corticotroph adenoma. AB - Silent corticotroph adenoma (SCA) is defined as an ACTH-producing pituitary tumor not associated with clinical and endocrine feartures of Cushing's syndrome, but its underlying molecular mechanism(s) remains unknown thus far. We tested the hypothesis that reduced expression of prohormone convertase (PC) 1/3 responsible for proteolytic processing of proopiomelanocortin (POMC) in SCA may lead to production of unprocessed, biologically inactive POMC and/or precursor of ACTH. Among 30 non-functioning pituitary macroadenomas (NFA) examined, we found 6 SCAs by immunohistochemical study using anti-ACTH antibody. Preoperative endocrine and diagnostic image tests did not reveal any differences between SCA and the remaining NFA except for the higher recurrence rate of SCA. While steady-state PC1/3 mRNA levels determined by RT-PCR were almost comparable between SCAs and NFAs, immunohistochemical study showed negative immunostaining for PC1/3 in all 6 SCAs. Our data suggest that defective PC1/3 expression may lead to preferential production of unprocessed, biologically inactive ACTH variants in SCA. PMID- 17917310 TI - Impaired autonomic function in patients with obstructive sleep apnea. PMID- 17917311 TI - Assessment of nocturnal blood pressure by home blood pressure monitoring. PMID- 17917312 TI - The proportion of individuals with alcohol-induced hypertension among total hypertensives in a general Japanese population: NIPPON DATA90. AB - Japanese men consume more alcoholic beverages than men in many other developed countries. The high consumption rate of alcoholic beverages among Japanese men may contribute to the high prevalence of hypertension in Japan. In the present study, we calculated the odds ratio for hypertension in alcohol drinkers based on recent criteria using data from a nationwide survey conducted in Japan in 1990, and estimated, among total hypertensives in a general Japanese population, the percentage of hypertensives whose condition was due to alcohol consumption. Of 3,454 male participants, 64.8% were drinkers (1 gou/day, 28.9%; 2 gou/day, 20.1%; 3 gou/day or more, 8.7%; ex-drinkers, 7.0%) and 49.8% were hypertensive, whereas 7.6% of 4,808 female participants were drinkers (1 gou/day, 5.2%; 2 gou/day or more, 1.3%; ex-drinkers, 1.1%) and 43.1% were hypertensive (1 gou=23.0 g of alcohol). In both sexes, drinkers had a higher odds ratio for hypertension than never drinkers, and there was a significant dose-response relationship between the amount of alcohol consumed and the odds ratio for hypertension. Among all hypertensives, the percentage whose hypertension was due to alcohol consumption was 34.5% (95% confidence interval, 10.9%-51.9%) for men and 2.6% (0.8%-5.8%) for women. The corresponding proportion based on daily alcohol intake was 12.7% for 1 gou/day, 11.1% for 2 gou/day, 5.8% for 3 gou/day or more, and 4.8% for ex drinkers in men, and 1.8% for 1 gou/day, 0.7% for 2 gou/day or more, and -0.1% for ex-drinkers in women. In conclusion, we found that a large percentage of the hypertensives in a general Japanese male population had alcohol-induced hypertension. PMID- 17917313 TI - Continuous positive airway pressure improves daytime baroreflex sensitivity and nitric oxide production in patients with moderate to severe obstructive sleep apnea syndrome. AB - Individuals with obstructive sleep apnea syndrome (OSAS) are at high risk for cardiovascular morbidity and mortality. The effects of OSAS severity and nocturnal continuous positive airway pressure (CPAP) on daytime baroreflex sensitivity (BRS) and nitric oxide (NO) production were investigated in OSAS patients. Fifty-one consecutive males with OSAS and 29 age-matched healthy men underwent the Valsalva test and standard polysomnography. Patients with an apnea hypopnea index (AHI) of >or=20 episodes per hour were randomized to receive CPAP treatment for 3 months (n=14) or no such treatment (n=19). The BRS index measured from the overshoot phase (phase IV) of the Valsalva maneuver and plasma NO concentration were significantly lower, whereas the AHI, oxygen desaturation time, arousal index, percentage of sleep stage 1, and systolic blood pressure were significantly greater, in patients with an AHI of >or=20/h than in those with an AHI of <20/h or in controls. The 24-h urinary excretion of norepinephrine was significantly reduced and the plasma NO concentration was significantly increased after one night of CPAP. The BRS index for phase IV and the Valsalva ratio were significantly increased in the CPAP group after the 3-month treatment period but remained unchanged in the non-CPAP group of OSAS patients. The daytime BRS index and NO production were thus inversely related to the severity of OSAS, and successful CPAP treatment improved these parameters in patients with moderate to severe OSAS. CPAP may therefore reduce the risk of cardiovascular complications due to endothelial dysfunction or increased sympathetic activity. PMID- 17917314 TI - Influence of hypertension on the incidence of cardiovascular disease in two rural communities in Japan: the Tanno-Sobetsu [corrected] study. AB - The purpose of this study was to determine the relationship between hypertension and onset of cardiovascular disease in Japan. As part of an ongoing epidemiological survey of cardiovascular diseases in Hokkaido, Japan, 1,798 subjects (806 males and 992 females; mean age in the initial year of the survey, 58.6+/-11.8 years) were selected, after excluding subjects who had been taking antihypertensive drugs, from a total of 2,136 subjects who had undergone medical examinations in 1991 in the town of Tanno and in 1992 in the town of Sobetsu, two rural communities in Hokkaido. Height, weight, casual systolic and diastolic blood pressures in the sitting position and blood biochemical values of all subjects were measured, and the subjects were divided into blood pressure level groups according to the 1999 World Health Organization/International Society of Hypertension (WHO/ISH) criteria. The follow-up survey was concluded at the end of August in 1999. The endpoints in this study were onset of circulatory disease or death due to circulatory disease. During the follow-up period, circulatory diseases (ischemic heart disease or stroke) occurred in 94 of the subjects. The incidence rates of cardiovascular disease (per 1,000 persons/year) for subjects divided into blood pressure groups according to the 1999 WHO/ISH blood pressure classification were 6.24 for the optimal+normal blood pressure level group, 11.26 for the normal high blood pressure level group, and 15.83 for the grade 1-3 hypertension group. Thus, the incidence rate of circulatory disease increased as the blood pressure level increased, and there was a significant difference between the incidence rate in subjects in the grade 1-3 hypertension group and the incidence rate in subjects in the optimal+normal blood pressure level group (p<0.05). In a Cox's proportional hazards model with onset of circulatory disease as the endpoint, diastolic blood pressure was shown to be an independent risk factor with a relative risk of 1.01. The results suggest that hypertension is an independent risk factor for onset of circulatory disease. PMID- 17917315 TI - Association study between hypertension and A/G polymorphism at codon 637 of the transporter associated with antigen processing 1 gene. AB - To explore the effect of A/G polymorphisms at codon 637 of the transporter associated with antigen processing 1 (TAP1) gene on the risk of hypertension. A case-control study of epidemiology was conducted. The case group included 277 community-based patients (136 males and 141 females; mean age 58.7+/-12.1 years) diagnosed with hypertension, and the control group consisted of 227 healthy subjects (95 males and 132 females; mean age 51.29+/-12.16 years) from the same community. The A/G polymorphisms at codon 637 of the TAP1 gene was examined by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method with genomic DNA. The effect of A/G polymorphisms at codon 637 of the TAP1 gene on hypertension was analyzed by using multivariate unconditional logistic regression models. The contribution of TAP1 637 A/G allele frequencies of the control group was consistent with that predicted by the Hardy-Weinberg equilibrium test (x2=230, p=0.632). There was a significant difference in the frequency of the A/G polymorphisms at codon 637 of the TAP1 gene between hypertensive patients (74.4/25.6%) and controls (82.4%/17.6%), x2=9.324, p=0.002. Genotype model (AA-AG-GG) analysis showed that there was a significant difference in the frequency of the recessive genotype between cases and controls (AA/AG vs. GG: odds ratio [OR]=3.046, 95% confidence interval [CI]=1.138-8.153) after adjustment for the covariates of age, serum total cholesterol, triglycerides, body mass index (BMI) and smoking. But there were no significant differences in the frequency of the genotype for the dominant model (AA vs. AG/GG: p=0.293) or additive model (AA vs. AG vs. GG: p=0.081) after adjustment. One-way ANOVA analysis showed that the systolic blood pressure, diastolic blood pressure, and BMI levels of the GG genotype were significantly higher than those of the AA or AG genotypes. In conclusion, our findings suggest that the A/G polymorphisms at codon 637 of the TAP1 gene contributes to the risk of hypertension, possibly via the increases in blood pressure and BMI. PMID- 17917316 TI - Comparison of the effects of efonidipine and amlodipine on aldosterone in patients with hypertension. AB - To prevent cardiovascular disease, targeting aldosterone synthesis and release may be clinically important. Aldosterone production in the adrenal gland is mediated mainly by the T-type calcium channel in vitro. Efonidipine inhibits both L- and T-type Ca channels. To compare the effects of efonidipine on neurohumoral factors with those of amlodipine, an L-type Ca channel blocker, we studied 40 essential hypertensive outpatients. Forty patients who had been administered amlodipine for more than 1 year were treated with efonidipine for 6 months in place of amlodipine. Substituting efonidipine for amlodipine had no significant effect on clinic systolic blood pressure or the plasma levels of brain natriuretic peptide, norepinephrine or active renin. However, the heart rate was significantly decreased (72.0+/-1.3 vs. 69.8+/-1.3 beats/min, p<0.01) and the plasma aldosterone level was also significantly decreased after efonidipine treatment (97.7+/-7.9 vs. 79.7+/-5.6 pg/mL, p<0.0001). Changes in the aldosterone level correlated with the baseline value before the replacement of amlodipine by efonidipine (r=-0.769, p<0.0001). These findings indicate that at the effective antihypertensive doses of efonidipine and amlodipine, efonidipine significantly decreases heart rate and plasma aldosterone level compared with those under amlodipine treatment in hypertensive patients. PMID- 17917317 TI - Angiotensin receptor blocker improves coronary flow velocity reserve in hypertensive patients: comparison with calcium channel blocker. AB - Large-scale clinical studies have indicated that angiotensin receptor blockers (ARBs) have beneficial effects against cardiovascular diseases. We designed this study to compare the effects of an ARB and a calcium channel blocker (CCB) on coronary flow velocity reserve (CFVR), a predictor of cardiovascular events, as estimated using transthoracic Doppler echocardiography. Sixteen hypertensive patients (63.1+/-9.6 years old; 10 males) were randomly allocated in a double blind fashion to valsartan (n=8, 40-80 mg/day) or nifedipine (n=8, 20-40 mg/day) groups. Age- and gender-matched subjects without hypertension were enrolled as a control group (n=12). CFVR was calculated by dividing the adenosine triphosphate induced hyperemic flow velocity by the basal flow velocity in the left anterior descending coronary artery. Baseline characteristics and reduction in systolic and diastolic blood pressure after 6 months were similar in both groups. CFVR in the valsartan group increased from 2.34+/-0.38 to 3.10+/-0.84 at 2 months (p<0.05), and to 3.04+/-1.09 at 6 months (p<0.01). Both values became comparable to that in the control group (2.81+/-0.60). CFVR in the valsartan group was significantly higher (p<0.001) than that in the nifedipine group, which was little changed at 6 months. This discrepancy was derived from the significant increase of hyperemic velocity in the valsartan group, from 36.6+/-17.3 cm/s to 41.1+/-12.7 cm/s at 2 months, and to 48.1+/-20.2 cm/s at 6 months. We concluded that the ARB valsartan not only reduced high blood pressure but improved CFVR in hypertensive patients. However, these effects were not seen with the CCB nifedipine. PMID- 17917318 TI - Reproducibility of nocturnal blood pressure assessed by self-measurement of blood pressure at home. AB - To assess the reproducibility of nocturnal blood pressure (BP) during sleep as measured using a self-measurement device at home, we obtained repeated nocturnal home BP at 0200 h and quality of sleep assessment from a diary in 556 subjects (71% women, 62.4+/-11.1 years) in the general population. We used an Omron device (HEM-747IC-N, Omron Healthcare Co., Ltd., Kyoto, Japan), with which the time and frequency of monitoring can be preset and the readings stored. The mean+/-SD of the difference between test-retest BP measurements was 0.7+/-15.1 mmHg systolic and 0.2+/-9.7 mmHg diastolic with a mean interval of 5.9 days. The absolute differences were greater than 10 mmHg in 261 (46.9%) subjects for systolic and 145 (26.0%) subjects for diastolic. There was no evidence of regression to the mean in nocturnal measurements over at least three nights (n=390, p>0.22). The differences (the first minus the second measurement) were large in subjects who experienced sleep disturbance only in the first (n=64, 2.3+/-13.6 mmHg and 1.6+/ 9.6 mmHg for systolic and diastolic, respectively) or second sessions (n=56, 4.1+/-16.4 mmHg and -2.5+/-11.4 mmHg) compared with the subjects without sleep disturbance (n=66, 1.5+/-17.8 mmHg and 0.8+/-10.3 mmHg) and those with sleep disturbance (n=370, 0.9+/-14.5 mmHg and 0.2+/-9.3 mmHg) in both sessions. In conclusion, the reproducibility of single nocturnal BP as assessed using a self measurement device at home was not good, especially for subjects who experienced different quality of sleep in each session. To evaluate nocturnal BP using a self measurement device, estimation of quality of sleep is indispensable. PMID- 17917319 TI - Limitation of the augmentation index for evaluating arterial stiffness. AB - Although the augmentation index (AIx) is widely used to evaluate arterial stiffness in clinics and research, some conflicting data exist in regard to its validity. We therefore performed a series of studies to test the validity of AIx. The first study in 196 peritoneal dialysis patients showed that AIx in diabetics was lower than that in non-diabetic patients (p<0.05), which was in contradiction with the previous studies. Further analysis showed that AIx was just weakly correlated with pulse pressure (PP)-a known index of arterial stiffness. We also found that the increase of augmentation pressure (AP) was usually accompanied with increased central PP (C-PP). As AP and C-PP are used as the numerator and denominator in the AIx formula, an increase in the numerator (AP) would not necessarily result in an increase of the quotient (AIx) unless the denominator (C PP) was stable. We then conducted a second study trying to test the validity of AIx through mathematical ratiocination. The increases in the central second peak (P2) and AP were assumed to represent increased arterial stiffness. Different values of AIx were obtained by varying the central initial systolic peak (P1) and diastolic pressure (DP). Mathematical ratiocination showed that AIx was dependent on multiple factors, F=(DeltaSP-DeltaDP)x(P1-P2)+(DeltaP2-DeltaP1)x(SP-DP), which suggested that a change of AIx would not always be attributable to changes in P2 and AP. This speculation was further proved by clinical data in our third study. In conclusion, through a series of studies and ratiocination, we showed that the augmentation index (AIx or AIx@75bpm) might not be a sensitive surrogate for a change in central pressure waveforms, which is a manifestation of change in large artery function. The limitation of AIx as an index of arterial stiffness is rooted in its formula, which has a clear mathematical flaw. PMID- 17917320 TI - Factors associated with baroreflex sensitivity: association with morning blood pressure. AB - Baroreflex sensitivity (BRS) is a primary mechanism for acute and chronic control of blood pressure (BP). However, there are few data showing the relationship between BRS and ambulatory BP (ABP). We assessed the hypothesis that BRS specifically contributes to some specific parameters of ABP in never-treated hypertensive/normotensive subjects. We studied 128 subjects (mean age: 54.5+/-13 years, 60% male) consisting of 92 untreated hypertensive and 36 normotensive subjects. Radial tonometric BP and simultaneous RR interval were recorded for 10 min, and the Valsalva maneuver was performed 3 times for each subject. BRS was calculated in two ways: the spontaneous-BRS by the spectral method, and the Valsalva-BRS by the slope method, using commercial software. ABP monitoring was performed on the same day as the BRS test. Of 128 subjects, we obtained BRS from 111 subjects with the Valsalva method and 123 subjects with the spontaneous method. Univariate analyses showed that the Valsalva-BRS was negatively correlated only with morning systolic BP (r=-0.21, p=0.03). Multivariable analyses showed that the Valsalva-BRS (ms/mmHg) was independently associated with the morning systolic BP (beta=-0.26, p=0.022), even after adjusting for age, sex, body mass index, presence of diabetes, duration of hypertension, clinic systolic BP and pulse rates. The spontaneous-BRS estimates were inversely correlated with clinic pulse rates in the same model. In conclusion, impaired BRS evaluated by the Valsalva method was associated with high morning BP level, independent of other confounders. Morning hypertension might be partly mediated by impaired BRS. PMID- 17917321 TI - Antihypertensive efficacy and safety of fixed-dose combination therapy with losartan plus hydrochlorothiazide in Japanese patients with essential hypertension. AB - A randomized, double-blind, placebo-controlled, parallel-group multicenter study was conducted to evaluate the antihypertensive efficacy and safety of 8-week treatment with one of three fixed-dose combinations-losartan 50 mg plus hydrochlorothiazide 12.5 mg, losartan 50 mg plus hydrochlorothiazide 6.25 mg, or losartan 25 mg plus hydrochlorothiazide 6.25 mg-in comparison with those of hydrochlorothiazide 12.5 mg alone, losartan 50 mg alone, or placebo in Japanese patients with essential hypertension. Significant reductions in sitting diastolic blood pressure (DBP) and systolic blood pressure (SBP) were seen in all three combination groups compared with the placebo group (each p<0.001). The greatest reductions in DBP and SBP were observed in the losartan 50 mg plus hydrochlorothiazide 12.5 mg group (12.7 and 18.0 mmHg, respectively). The reductions in the losartan 50 mg plus hydrochlorothiazide 12.5 mg group were significantly greater (each p<0.001) than those in the placebo group and each of the monotherapy groups. There were no significant differences in the incidences of clinical and laboratory drug-related adverse events between any of the combination groups and the placebo group. All combination groups showed improved hypokalemia and hyperuricemia compared to the hydrochlorothiazide 12.5 mg group. These results demonstrated that once-daily, fixed-dose combination therapy with losartan 50 mg plus hydrochlorothiazide 12.5 mg is well tolerated and more efficacious in lowering DBP and SBP than monotherapy in Japanese hypertensive patients. PMID- 17917322 TI - Ability of ambulatory blood pressure monitoring and myocardial/carotid ultrasound to predict the location and the severity of coronary artery lesions in normotensive patients: a clinical study. AB - Pulse pressure has been recognized as a marker of cardiovascular disease in normotensives. Moreover, internal carotid artery intima-media thickness (IMT) has been proposed to reflect coronary artery lesions. The aim of the present study was to evaluate the predictive value of other parameters derived from ambulatory blood pressure monitoring (ABPM), myocardial ultrasound, and carotid ultrasound to predict the location and the severity of coronary artery disease in normotensives. One hundred and thirteen patients with suspected coronary artery disease underwent coronary angiography, 24-h ABPM and myocardial/carotid ultrasound. Multivariate analysis was applied and equations were extrapolated based on independent variables derived from ABPM and ultrasound. The Gensini score was independently correlated with male gender, pulse pressure, average heart rate for both 24-h (p=0.001) and night (p=0.006) values, as well as percentage of high systolic blood pressure (BP), average diastolic BP, average mean BP, and heart rate concerning daily mesurements (p=0.001). Moreover, the Gensini score was independently correlated with end-systolic volume, posterior wall thickness during systole and intraventricular septum thickness during diastole, along with male gender and age (p=0.001), as well as mean internal and right common carotid artery IMT (p=0.002). Similar mathematical formulas have been calculated separately for the coronary arteries and their main branches. In conclusion, the location and the severity of coronary disease can be effectively evaluated by ABPM and myocardial/carotid ultrasound in normotensives. This approach could be useful for determining atypical patients at risk and/or for treating patients with suspected coronary disease who refuse coronary angiography. PMID- 17917323 TI - The contribution of nutrition to the protective value of high plasma aldosterone concentrations in hemodialysis patients. PMID- 17917325 TI - Serum butyrylcholinesterase is strongly associated with adiposity, the serum lipid profile and insulin resistance. AB - OBJECTIVE: We investigated the association between the serum butyrylcholinesterase (BChE) activity and parameters of adiposity, insulin resistance and the serum lipid profile, including the serum levels of triglyceride (TG), HDL-cholesterol (HDL-C) and LDL-cholesterol (LDL-C) in Japanese patients with type 2 diabetes and non-diabetic subjects. PATIENTS AND METHODS: A total of 259 Japanese subjects [171 patients with type 2 diabetes (age 60.4+/-11.9 years, BMI 25.0+/-4.4) and 88 non-diabetic subjects (age 54.2+/-15.0 years, BMI 25.8+/-4.7)] were enrolled in the study. To assess the hepatic fat content, the ratio of the CT attenuation value of the liver to that of the spleen (L/S ratio) was calculated. RESULTS: Serum BChE activity was significantly correlated with various indices of adiposity, including the waist circumference, L/S ratio, visceral fat area (VFA), subcutaneous fat area (SFA) and BMI, and also with the TG, HDL-C, LDL-C and HOMA-R, in the entire subject population. Even after adjustment for five variables, namely, age, gender, HbA1c, CRP and serum ferritin (the serum levels of ferritin and CRP have been reported to be associated with adiposity and insulin resistance), the serum BChE activity was still correlated with the waist circumference, L/S ratio, VFA, SFA, BMI, TG, HDL C, LDL-C and HOMA-R. Furthermore, changes in the serum BChE activity were associated with changes in the L/S ratio, VFA and SFA at 1 year after the baseline. CONCLUSIONS: These results indicate that the serum BChE activity is associated with parameters of adiposity, the serum lipid profile and the degree of insulin resistance. PMID- 17917326 TI - A retrospective study on the efficacy of corticosteroid-alone therapy in membranous nephropathy patients. AB - OBJECTIVE: Membranous nephropathy (MN) is the most common cause of adult-onset nephrotic syndrome and its management is still controversial. The aim of this study was to determine the effectiveness of corticosteroid-alone therapy for controlling proteinuria in MN. METHODS: Twenty-three patients, which had moderate proteinuria (admission 24-hour urinary protein excretion 1.0 to 3.5 g/day) with primary MN were studied retrospectively. RESULTS: Thirteen patients received corticosteroid-alone therapy combined with rest and dietary therapy (steroid group), while the other 10 patients were treated with rest and diet alone (non steroid group). These two groups did not differ with respect to their laboratory features at the time of admission. After discharge, 5 of 13 patients of the steroid group dropped out . Therefore, only 8 patients could be followed up. As the result, 5 of 8 patients (62.5%) achieved complete remission (CR) and 3 of 8 patients (37.5%) had incomplete remission (ICR), so none of the patients failed to improve. On the other hand, 3 of 10 patients of the non-steroid group dropped out. Then, 7 patients were followed up. None of the 7 patients showed improvement during follow-up and 5 of these 7 patients were started on corticosteroids. Finally, as this result, 4 of 5 patients (80%) could achieve CR by 2 years after hospital discharge. Moreover, in the remaining 2 patients from the non-steroid group, no remission could be achieved even 2 years after discharge. CONCLUSION: These results suggest that long-term corticosteroid-alone therapy is beneficial for controlling proteinuria in patients with MN. PMID- 17917327 TI - A large inflammatory fibroid polyp in the sigmoid colon treated by endoscopic resection. AB - Inflammatory fibroid polyp (IFP) is a rare, localized, nonneoplastic lesion originating from the submucosa of the gastrointestinal tract. Microscopically, these lesions are made up of a complex network of variable-size blood vessels and diffuse inflammatory cells contained in the edematous stroma. They are most often found in the stomach, followed by the small intestine, and rarely in the esophagus or the large intestine. We report a case of sigmoid colonic IFP presenting bloody stool treated with endoscopic resection. PMID- 17917328 TI - Significance of an epidermal growth factor receptor mutation in cerebrospinal fluid for carcinomatous meningitis. AB - We report a case of epidermal growth factor receptor (EGFR) inhibitor-sensitive lung adenocarcinoma with carcinomatous meningitis who showed a good response to gefitinib, an oral tyrosine kinase inhibitor of EGFR. This good response to gefitinib treatment was attributed to evidence of an EGFR mutation, L858R in exon 21, which was detected in a small amount of cerebrospinal fluid (CSF) before the positive CSF cytology. Patients with carcinomatous meningitis often have a poor performance status, and therefore diagnostic approaches and therapeutic methods are also often limited. Detection of EGFR mutations may be a useful method for non-small cell lung cancer diagnosis, and also facilitate determination of appropriate therapeutic protocols. PMID- 17917329 TI - Sarcoidosis accompanied by systemic lupus erythematosus and autoimmune hepatitis. AB - A 52-year-old woman was admitted to our hospital for further examination of blurred vision, abnormal lung shadows and an elevated level of angiotensin converting enzyme. Sarcoidosis was suspected, however, careful history taking revealed the existence of photosensitivity and polyarthralgia. Laboratory tests showed lymphocytopenia, liver dysfunction, hypergammaglobulinemia, and positive anti-nuclear, anti-double stranded DNA and anti-smooth muscle antibodies. Liver biopsy examination showed chronic active hepatitis. She was diagnosed with the triplex of sarcoidosis, systemic lupus erythematosus and autoimmune hepatitis. Marked improvement was noted after corticosteroid therapy. PMID- 17917330 TI - Pulmonary endarteritis and subsequent pulmonary embolism associated with clinically silent patent ductus arteriosus. AB - A 49-year-old man without heart murmur was admitted with fever because of bacteremia following a tooth extraction. Antibiotics rapidly alleviated the fever; however, a small nodule in the pulmonary artery was identified on computed tomography (CT). When the patient experienced chest discomfort with fever, CT demonstrated the absence of the nodule and the appearance of an abnormal lung opacity, and echocardiography showed turbulent retrograde flow in the pulmonary artery. We had the rare opportunity to follow a case of pulmonary bacterial endarteritis and subsequent pulmonary embolism with clinically silent patent ductus arteriosus (PDA) that was confirmed by 3-dimensional CT. PMID- 17917332 TI - Intermittent intravenous immunoglobulin successfully prevents relapses of neuromyelitis optica. PMID- 17917331 TI - Evaluation of platelet transfusion thresholds in patients with acute myeloblastic leukemia receiving induction chemotherapy. PMID- 17917333 TI - Hydatid cysts: three different stages on the same image. PMID- 17917334 TI - Tumor mimicking hepatic echinococcus alveolaris with portal vein thrombosis. PMID- 17917336 TI - [Effect on image data resampling in evaluation of the basic imaging properties for a digital radiographic system based on a flat panel detector]. AB - We investigated the effect on image data resampling in an evaluation of the basic imaging properties for a digital radiographic system based on a flat panel detector (FPD). One of the latest digital radiographic systems was used in this study. This system was based on a direct-conversion FPD of amorphous selenium. The basic imaging properties of the system were evaluated by measuring characteristic curve, presampled modulation transfer function (MTF), and Wiener spectrum (WS) using DICOM image with a matrix size of 2048 x 2048. The evaluations were performed under two conditions because matrix size automatically changes according to the selection of imaging size. One of the conditions was a different matrix size between image data acquired on the FPD and the output image (DICOM image for which resampling was performed). The other condition was that these matrices be the same size (DICOM image with no resampling performed). Resampling did not affect the characteristic curves. However, MTF and the WS obtained from the resampled data were different from those of the one not resampled, which is considered to be the "inherent" basic imaging properties, and this phenomenon was remarkable, especially in terms of the MTFs. Our study indicates that the effect on resampling should not be disregarded in evaluating the basic imaging properties of digital radiographic systems. Therefore, it is mandatory to use DICOM images for which no resampling was performed in order to evaluate the inherent basic imaging properties for digital radiographic systems. PMID- 17917335 TI - Synchronous small and non-small cell lung cancer in a patient with previous tuberculosis. PMID- 17917337 TI - [Feasibility of diagnostic contrast-enhanced CT for attenuation correction of whole body PET images]. AB - OBJECTIVE: Image fusion has been recognized as a useful technique in diagnostic imaging. We have been evaluating the manual image fusion of PET and contrast enhanced (CE) CT obtained separately. The CT images can be used for attenuation correction as well as for image fusion; however, the quantitative accuracy of CT corrected PET images has yet to be assessed. The purpose of this study was to compare the radioactivity concentration between conventional (68)Ge-corrected and CECT-corrected PET images. METHODS: Twenty patients underwent a whole-body PET scan, followed by a CT scan with intravenous contrast material, after careful positioning using an individually molded vacuum cushion. Two different attenuation-corrected emission data sets were produced, i.e., (68)Ge-corrected images and CECT-corrected images. Image registration was performed by maximizing mutual information-based cost function, between the CT and the combination of emission and transmission PET volumes. The CT pixel values in Hounsfield units were transformed into linear attenuation coefficients in cm(-1), using a conversion formula for a lookup-table from phantom experiments. Measured activity concentrations from identical regions of interest in representative normal organs and in 25 pathologic foci of uptake were compared. In addition, the quality of the reconstructed images was assessed using the normal mean square error (NMSE). RESULTS: Measured average radioactivity concentrations were 1.38-9.56% higher for CECT-corrected images than for (68)Ge-corrected images. Overall, the NMSE value of CECT-corrected images compared with (68)Ge-corrected images was 0.02+/-0.01. CONCLUSIONS: The difference in quantitative values between (68)Ge-corrected and CECT-corrected PET images was comparable to that of an integrated PET/CT system. Diagnostic CT images with intravenous contrast performed separately before or after a PET scan could be used clinically not only for fusion but also for attenuation correction. PMID- 17917338 TI - [Testing of dynamic multileaf collimator by dynamic log file]. AB - Intensity-modulated radiation therapy (IMRT) represents one of the most significant technical advances in radiation therapy. In the dynamic multileaf collimator (MLC) method of IMRT delivery, because of the relatively small gaps between opposed leaves and because most regions are shielded by leaves most of the time, the delivered dose is very sensitive to MLC leaf positional accuracy. A variation of +/-0.2 mm in the gap width can result in a dose variation of +/-3% for each clinical dynamic MLC field. Most often the effects of leaf motion are inferred from dose deviations on film or from variations in ionization measurements. These techniques provide dosimetric information but do not provide detailed information for diagnosing delivery problems. Therefore, a dynamic log file (Dynalog file) was used to verify dynamic MLC leaf positional accuracy. Measuring for narrow gaps using the thickness gauge could detect a log file accuracy of approximately 0.1 mm. The accuracy of dynamic MLC delivery depends on the accuracy with which the velocity of each leaf is controlled. We studied the relationship between leaf positional accuracy and leaf velocity. Leaf velocity of 0.7 cm/sec caused approximately 0.2 mm leaf positional variation. We then analyzed leaf positional accuracy for the clinical dynamic MLC field using Dynalog File Viewer (Varian Medical Systems, Inc., Palo Alto, CA), and developed a new program that can analyze more detailed leaf motions. Using this program, we can obtain more detailed information, and therefore can determine the source of dose uncertainties for the dynamic MLC field. PMID- 17917339 TI - [Examination of tumor diameter measurement precision by RECIST]. AB - Image evaluation with Response Evaluation Criteria in Solid Tumors (RECIST) evaluates the change in a measurable lesion as determined by ruler or micrometer caliper. However, there is no definition of the conditions thought to influence the precision of measurement. We therefore examined the effects on measurement precision by changing image amplification, WW, WL, and time phase. Moreover, to determine response rate, one-dimensional evaluation with RECIST was compared with the two-dimensional evaluation of World Health Organization (WHO) for a hepatocellular carcinoma. The results of measuring the object lesion for measured value variation were as follows. Under image conditions of 1 time expansion/ WW 150/ WL 100 was (4.92+/-1.94)%. Under image conditions of 1 time expansion/ WW 350/ WL 75 was (4.42+/-1.70)%. Under image conditions of 4 times expansion/ WW 150/ WL 100 was (2.52+/-0.82)%. Under image conditions of 4 times expansion/ WW 350/ WL 75 was (2.83+/-1.10)%. When an image was enlarged to 4 times, precision doubled. There was no a difference in comparing RECIST to WHO in terms of response rate. Thus the best method was considered to be RECIST because of its convenience. PMID- 17917340 TI - [The large intestine: examination methods and colon tumors]. PMID- 17917341 TI - [Cost utility analysis and quality of life]. PMID- 17917342 TI - [How to adjust the MRI parameters]. PMID- 17917343 TI - [How the Japanese civil code lawyers see the patient-medical staff relationship? A gap of medical and legal reality]. PMID- 17917346 TI - [Safety support for medical device: an outline revision and highlights of the medical law]. PMID- 17917344 TI - [A study of the guideline of MDCT-imaging method for lung cancer screening]. PMID- 17917347 TI - [Development of an algorithm for the detection of early signs of cerebral ischemia on CT images]. AB - This paper describes an approach to automatically detect the parenchymal hypoattenuation of hyperacute stroke in nonenhanced computed tomography (CT) images. This technique is based on a comparison between the CT values histograms of right and left brains. A subtraction curve that was regarded as an output value was calculated by subtracting the right-hemispherical histogram from the left-hemispherical histogram obtained from one of the region-of-interest (ROI) sets on an image. The output value was used to assess whether hypoattenuation exists on CT images with a threshold value. If judged abnormal, a rectangular region including a whole or partial hypoattenuation area was detected from the ROI. Twenty-six cases with hypoattenuation and 30 cases without hypoattenuation were included in this study. As a result of our experiments, the sensitivity of this method in detecting hypoattenuation was found to be 92%, with approximately 0.16 false-positive per image. Our preliminary experimental results indicated that the proposed technique can be used for the automated detection of parenchymal hypoattenuation of hyperacute stroke on nonenhanced CT images. PMID- 17917348 TI - [Bone fusion algorithm: a new tool to support decision making]. AB - Three dimensional angio (3D-Angio) is indispensable in neuroradiology, especially for examinations and interventions of cerebral-aneurysms. 3D-Angio has two modes, 3D-DSA and 3D-DA. The former mode is used to determine vessel structures in detail, and the latter is good at simultaneously determining relationships between vascular and osseous structures. However, it is sometimes difficult to determine these relationships with 3D-Angio if they are close together. The relationship between the skull base and adjacent aneurysm is an essential factor in determining treatment strategy. In order to make this determination, we have developed the Bone Fusion function, which reconstructs vessel and osseous structures separately from rotational DSA data and its mask sequence data, respectively, and two independent 3D data sets are fused in one 3D representation. There are three display modes: fusion, vessel only, and bone only, and those structures are clearly identified by different colors on the fusion images. The Bone Fusion function was applied to hundreds of clinical cases in which 3D-DSA and 3D-DA were used. The Bone Fusion provided important information to determine treatment strategy, although those factors were sometimes lost with 3D-DSA and 3D-DA when the studied vasculatures were adjacent to bone. It was especially useful to apply the Bone Fusion function to aneurysms or tumors adjacent to the skull base. PMID- 17917349 TI - [Estimation of personal dose based on the dependent calibration of personal dosimeters in interventional radiology]. AB - The purpose of present study is, in interventional radiology (IVR), to elucidate the differences between each personal dosimeter, and the dependences and calibrations of area or personal dose by measurement with electronic dosimeters in particular. We compare space dose rate distributions measured by an ionization survey meter with the value measured by personal dosimeter: an optically stimulated luminescence, two fluoroglass, and two electronic dosimeters. Furthermore, with electronic dosimeters, we first measured dose rate, energy, and directional dependences. Secondly, we calibrated the dose rate measured by electronic dosimeters with the results, and estimated these methods with coefficient of determination and Akaike's Information Criterion (AIC). The results, especially in electronic dosimeters, revealed that the dose rate measured fell by energy and directional dependences. In terms of methods of calibration, the method is sufficient for energy dependence, but not for directional dependence, because of the lack of stable calibration. This improvement poses a question for the future. The study suggested that these dependences of the personal dosimeter must be considered when area or personal dose is estimated in IVR. PMID- 17917350 TI - [Method of segmenting inferior horns of lateral ventricles using active contour models]. AB - Recent research has suggested that the measurement of regional atrophy in the structure of the medial temporal lobe is a promising way to discriminate Alzheimer-type dementia patients from healthy control subjects. There are some reports that the inferior horns of the lateral ventricles are expanded by atrophying the structure of the medial temporal lobe. We developed a technique to automatically detect the region of the inferior horns of the lateral ventricles by gray-level thresholding and morphological processing. However, there were some incorrect regions in this method. Accordingly, we proposed a technique for which active contour models (ACM) were used. Our ACM incorporates the improved edge based image and the external constraint to improve convergence and to reduce its dependence on initial estimation. In this study, we present the details of an algorithm that traces the contours of the inferior horns of the lateral ventricles and its performance relative to manual methods. The average degree of correspondence between the extract region and manual trace was measured in 30 inferior horns of 15 subjects. The average degree of correspondence of the proposed method was about 4% higher than that of the conventional method. These results suggest that the proposed method is more accurate than the conventional method. PMID- 17917351 TI - [Method of calculating patient skin surface dose in transcatheter arterial embolization]. AB - The usefulness of interventional radiology (IVR) in clinical practice is well known. However, patient dose in IVR has recently been increased as a result of the prolongation of fluoroscopic time and the increased number of radiographies. We studied a simple method of calculating skin surface dose in patients who underwent transcatheter arterial embolization (TAE) for the treatment of hepatocellular carcinoma by obtaining the value of a dose area product meter attached to the digital subtraction angiography system. In 20 subjects (15 men and 5 women, aged an average of 68.2+/-7.3 years, respectively) who underwent TAE, exposure conditions (tube voltage, tube current, time, and size of image intensifier) in a time series and last value indicated on the dose area product meter were recorded. A dosimetric phantom was placed at a position the same as that of the patient for TAE, the surface dose (SD) of the phantom was measured under various exposure conditions, and SD per unit mAs (SD/mAs) was obtained. Then the skin surface dose in each subject was estimated from the values of the exposure condition and SD/mAs. A high correlation was observed between the last value (x) on the dose area product meter and the estimated skin surface dose (y) (r=0.933), and the following regression equation was derived: y=0.005x-0.589. The skin surface dose calculated using the regression equation was compared with that obtained by the method recommended by the Japan Association on Radiological Protection in Medicine (JARPM), considering the value estimated from the value of exposure conditions with SD/mAs as the gold standard. The results indicated that the error in the method using the regression equation was significantly lower than that of the JARPM method (18.3+/-14.0% and 75.5+/-66.0%, respectively, p<0.01). In conclusion, the skin surface dose in TAE could be monitored with high precision using the value of the dose area product meter by obtaining the regression formula between the value of the dose area product meter and the skin surface dose estimated with the phantom values. PMID- 17917352 TI - [Evaluation of the accuracy of a compensating filter based on compact NC-Mill for radiation therapy]. AB - The present report describes the fabrication technique and dosimetry aspects of a desktop numerically controlled milling machine (NC-Mill) based a compensator system that uses lead clay (Shield cray, Reactor Experiments, Inc., U.S.A.). Effective path lengths of patients were determined for CT image sets using the ray-tracing technique and converted to compensator thickness with the equivalent TMR method. Rigid urethane foam was processed with the NC-Mill to produce a mold for filters, and the lead clay was adopted as the compensating material. The dose distribution was measured on the compensating plane of an anthropomorphic phantom and a stair-step PMMA phantom. It was found that the radiation field with inhomogeneous dose was as high as 30%+/-3% with the compensating filters. In addition, when the absorbed dose at the central axis of 52 compensating filters that were used clinically was measured, 75.0% showed an error of less than +/-3%, and 3.8% showed the maximum dose error: >+/-5%. Overall, the present system was capable of producing dose uniformity to within +/-5% for a stair-step phantom, an anthropomorphic phantom, and clinical situations. PMID- 17917353 TI - [Investigation of a CT value electron density conversion method in radiotherapy planning CT]. AB - To obtain heterogeneous corrections, it is necessary to convert computed tomography (CT) values into electron density. However, differences in the current conversion tables of the method of acquisition for each institution are not clear. Therefore, we sent a questionnaire survey to 200 radiotherapy institutions in the Kanto area regarding heterogeneous correction data acquisition. When these conversion tables were created, many institutions used phantoms for conversion, and 85% of institutions borrowed them from the factories. On the other hand, some institutions used the factory values as the other conversion method. In the conversion tables of the various institutions, we recognized large differences between institutions, in that relative electronic density (Rel.ED) was greater than 1.0. When we assumed the CT value of bone to be 793 HU, the mean relative electron density at 120 kV was 1.46+/-0.09, with a minimum of 1.36 and maximum of 1.82. We have recognized the need to create guidelines detailing these methods of evaluation. PMID- 17917354 TI - [Technological study of image quality in PET examination using commercial FDG delivery]. PMID- 17917355 TI - [The large intestine: inflammatory bowel disease (IBD)]. PMID- 17917356 TI - [Evidence based medicine and clinical economics]. PMID- 17917357 TI - [Dosimetry of absorbed dose in clinical proton beams]. PMID- 17917358 TI - [The Japanese Supreme Court dealt with a Judo therapist case: a case study of violation of medical qualification laws]. PMID- 17917359 TI - [The Amendment of the Medical Service Law for ensuring the medical safety]. PMID- 17917360 TI - [Safety support for medical device: 2. Concrete contents of revision and points to be considered]. PMID- 17917362 TI - [Study of susceptibility-weighted imaging (SWI) using a simple MR phantom]. AB - We evaluated the visibility of hypointensity regions on susceptibility-weighted (SW) magnetic resonance imaging (MRI). A commercial simple phantom filled uniformly with a gel material was demonstrated to include small regions affected by different magnetic susceptibilities compared to their surroundings. For detection of these regions in the phantom, the three-dimensional SW imaging (SWI) technique is superior to a conventional two-dimensional gradient-recalled-echo (GRE) MRI technique. The mean contrast between the hypointensity regions and their surroundings on GRE images (T2* weighted images) of 4 mm slice thickness is approximately 88% less than that on SWI of 4 mm effective slice thickness. When the effective slice thickness of SWI is increased more than 4 mm, the contrast on SW images is decreased. While the mean contrast on SWI of 7 mm effective slice thickness is approximately 75-65% compared to that of 4 mm effective slice thickness, its contrast of 7 mm is determined to be higher than that on GRE images of 4 mm slice thickness; this suggests that the SWI technique could be applied to whole brain examination by reducing the acquisition time. The quantitative results in this article are considered to be useful for evaluating the visibility of hypointensity regions on SWI, when comparing them with GRE images and varying the effective slice thickness of SWI. PMID- 17917365 TI - [Safety support for medical device: 3. Business: what we should do for control]. PMID- 17917363 TI - [Method of SNR determination using clinical images]. AB - Parallel magnetic resonance imaging (MRI) with arrays of receiver coils such as those of sensitivity encoding (SENSE) are being widely used. However, conventional methods of image signal-to-noise ratio (SNR) determination cannot be used for parallel MRI, and a novel method has been reported in JSRT. However, this method of SNR determination is for phantom images not for clinical images. Therefore, we researched accurate measurement of the image noise of parallel MRI reconstruction including the unfolding process and uniformity filters. The possibility of a subtraction method using clinical images and the accuracy of standard deviation (SD) of clinical images for optimum ROI were studied because it was not possible to use the air-signal method. The results indicated that the position of the ROI was selected for uniformity of signal intensity area and that the size of the ROI was about 50 pixels. However, under these conditions, the noise value of SNR was higher than that using the phantom-subtraction method. In addition, the tissue-subtraction method was useful when the two scanning images were in agreement. PMID- 17917366 TI - Regional differences of L-type high voltage-gated calcium channel subunit expression in the mouse brain after chronic morphine treatment. AB - As functional changes in L-type high voltage-gated calcium channels (HVCCs) are recognized to be one of the major neurochemical modifications occurring in brains of animals with morphine physical dependence, this study attempts to examine whether regional difference in the expressions of HVCC subunits are produced in the brains under such pathological conditions. Scatchard analysis of [(3)H]PN200 110 binding showed increased Bmax values in the cerebral cortex and the mesolimbic region including the nucleus accumbence, which are brain regions participating in the development of morphine physical dependence, but not in the cerebellum. In the former two brain regions, alpha1C and alpha1D subunits of L type HVCCs and alpha2/delta1 subunit increased, although decreases of alpha1B and alpha2/delta1 subunits were observed in the cerebellum. A single dose of morphine did not change the expression of any of the HVCC subunits. These results indicate that the increased L-type HVCC subunits in the cerebral cortex and mesolimbic region participate in the development of morphine physical dependence. PMID- 17917368 TI - Physical and histopathological assessment of the effects of metallic stents on radiation therapy. AB - To evaluate whether simultaneous metallic stent (MS) placement and radiotherapy are feasible, phantom and animal experiments were performed. The interface dose by external irradiation (EI) or intracavity irradiation (II) to 5 kinds of MS was measured using the charge-coupled device (CCD) camera with a thermoluminescent (TL) sheet, and backscatter and absorption were evaluated using composite method. Lineac 10 MV X-ray irradiated the MS in close contact with the TL sheet. II was performed using (192)Ir, and the irradiation dose transmitted through the MS was measured using the TL sheet. The ratio of the CCD value of the MS wire region to that of the MS non-wire region was defined as the dose perturbation factor (DPF). Furthermore, the effects of a combination of (60)Co gamma-ray EI and MS placement in the normal common bile duct were histopathologically evaluated in dogs. In the phantom experiments of EI, in backscatter by the MS, the DPF was 1.09 for CZ, and 1.03 for Pal, but no backscatter was detected in the remaining 3 MS. In absorption by the MS, the DPF was 0.92, 0.97, 0.97, and 0.98 for CZ, Wall, Pal, and Vel, respectively, but no absorption was detected in U. Flex. In those of II, the DPF of absorption was 0.91, 0.98, and 0.98 for CZ, U. Flex, and Wall, respectively, but no absorption was detected in Pal and Vel. The animal experiments showed infiltration of inflammatory cells and fibrosis in the case of both MS placement and EI. These changes were marked in EI treating after MS placement, but neither severe ulcer nor perforation was found. In conclusion, these results suggested that the effect of MS should be considered carefully when simultaneous MS placement and EI is performed clinically. PMID- 17917367 TI - Effects of pioglitazone on increases in visceral fat accumulation and oxidative stress in spontaneously hypertensive hyperlipidemic rats fed a high-fat diet and sucrose solution. AB - We examined oxidative stress and metabolic characteristics of the spontaneously hypertensive hyperlipidemic rat (SHHR) when it was fed a high-fat diet and sucrose solution (HFDS) after N(G)-nitro-L-arginine methyl ester ingestion to develop a rat model of metabolic syndrome. This study was carried out to assess the effects of pioglitazone on levels of lipid peroxide (LPO), Cu,Zn superoxide dismutase (Cu,Zn-SOD), catalase (CAT), glutathione peroxidase (GPx), and non esterified fatty acids (NEFA) in the plasma and liver tissue in HFDS-SHHR compared with Sprague-Dawley rats (SD). In the HFDS-treated groups, levels of LPO, CAT, GPx, and NEFA were elevated and levels of Cu,Zn-SOD were reduced in the plasma and liver tissue, with a marked accumulation of visceral fat. The changes induced by HFDS feeding were severe in the SHHR model that had essential hypertension and hyperlipidemia, when compared with SD that did not have these essential risk factors. Subcutaneous injection of 10 mg/kg per day of pioglitazone for 2 months significantly restored levels of LPO, CAT, GPx, Cu,Zn SOD, and NEFA in the HFDS-SHHR group, and visceral fat accumulation was reduced. These results suggest that HFDS-SHHR is a suitable model of metabolic syndrome and that pioglitazone treatment can improve oxidative dysregulation in this rat model. PMID- 17917369 TI - An extract of Phyllanthus amarus protects mouse chromosomes and intestine from radiation induced damages. AB - We reported earlier on our preliminary study of the radioprotective effect of Phyllanthus amarus (P.amarus) in mice. P.amarus was found to inhibit the myelosuppression and elevated the levels of antioxidant enzymes in the blood and liver. In the present study we have evaluated the protective effect of P.amarus against radiation-induced changes in the intestine and mouse chromosomal damage. P.amarus at concentrations of 250 & 750 mg/Kg. b. wt were found to elevate the antioxidant enzymes in the intestine and decrease the lipid peroxidation levels. Histopathological evaluations of the intestine revealed decreased damage to intestinal cells, demonstrating that P.amarus protected the intestine. The genotoxic effects of radiation on mouse chromosomes were evaluated by assaying the micronuclei formation and chromosomal aberrations. P.amarus was found to protect the clastogenic effects of radiation as seen from decreased number of micronuclei. The administration of P.amarus was also found to decrease the percentage of chromosomal aberrations. Based on our present and previous reports it could be concluded that P.amarus extract has significant radioprotective activity. PMID- 17917370 TI - Streptozotocin-induced diabetes decreases placenta growth factor (PlGF) levels in rat placenta. AB - The placenta produces several growth factors, including placenta growth factor (PlGF), which are essential for placenta growth and fetal growth. Diabetic pregnancy induces the abnormal placental growth and fetal development. This study investigated whether diabetes in pregnant rats induces changes in PlGF expression in the placenta. Diabetes was induced by a single intravenous injection of streptozotocin (35 mg/kg body weight) on day 0 of pregnancy, blood and tissue samples were collected on day 20 of pregnancy. In the diabetic group, maternal body weight and fetal weight significantly decreased compared to controls. RT-PCR and Western blot analyses showed that expression of PlGF was significantly decreased in placenta by streptozotocin treatment. Immunohistochemical study showed that the positive signal of PlGF in trophoblast cells was decreased in the diabetic group compared to controls. These findings demonstrate the decline of PlGF in the placenta in diabetic pregnancy. PMID- 17917371 TI - Adenosine deaminase activity in cats infected with feline immunodeficiency virus. AB - Adenosine deaminase (ADA), an enzyme involved in purine metabolism, has been shown to be of clinical importance in several diseases in humans. To investigate whether ADA is of any clinical significance in cats, plasma adenosine deaminase (P-ADA) and T cell adenosine deaminase (T-ADA) activities were measured in feline immunodeficiency virus (FIV) negative and positive cats. The AIDS-related complex (ARC) group showed a significant elevation in P-ADA activity compared to the asymptomatic carrier (AC), and FIV-negative groups (P<0.005). T-ADA activity was significantly elevated in FIV-positive cats compared to the FIV-negative group (P<0.05) and this elevation was attributed to the increase in the ARC group (P<0.01). A correlation was found between P-ADA and T-ADA activities in the FIV negative group. T-ADA activity and CD4(+)cell number showed a strong negative correlation in FIV-positive cats (P<0.0005). CD4(+) cell numbers were significantly reduced in the ARC group compared to the healthy controls (P<0.005). Our results showed that T-ADA is increased in FIV-positive cats during the ARC stage. These results also suggest that ADA may be an indicator of T cell activation in the ARC stage of FIV infection. PMID- 17917372 TI - Cyclosporine A inhibits the mRNA expressions of IL-2, IL-4 and IFN-gamma, but not TNF-alpha, in canine mononuclear cells. AB - The effects of the calcineurin inhibitors cyclosporine A (CsA) and FK506 on the mRNA expressions of various cytokines were evaluated in dogs to determine whether the effects of CsA and FK506 in dogs were similar to those in humans. The mRNA expression levels of the cytokines IL-2, IL-4, IFN-gamma and TNF-alpha were measured in PHA-stimulated canine PBMC using real-time RT-PCR after incubation with CsA or FK506 for 5 hr. Both reagents inhibited IL-2, IL-4 and IFN-gamma mRNA expressions in a dose-dependent manner. However, CsA hardly inhibited the mRNA expression of TNF-alpha. These findings are important for assessing the indications of CsA treatment in dogs. PMID- 17917373 TI - Controlling highly prevalent Staphylococcus aureus mastitis from the dairy farm. AB - In 57 Holstein cows where the dairy farm uses a milking parlor system, the somatic cell count (SCC) increased persistently in the bulk milk (monthly mean 52.3 x 10(4) cells/ml; range 21 to 94 x 10(4) cells/ml). We detected S. aureus in 24 (41.2%) of the 54 lactating cows and in 29 (12.8%) of 227 quarters of the 57 milking cows in the herd. A control program was implemented in an effort to eradicate S. aureus mastitis from this dairy farm. The control plan established improved handling of the lactating cows, improved milking procedures, dry-cow therapy, and culling of infected cows. The program was monitored for 3.5 years by frequent checkups on the rate of S. aureus infection, the SCC, and the changes in milk composition. Eighteen months after the control program was started, the rate of S. aureus infection in the quarter milk decreased dramatically, and no S. aureus isolates were found in the milk of the remaining cows. The SCC in the bulk milk of the herd dropped to a monthly mean of <20 x 10(4) cells/ml. In conclusion, the control program was effective for controlling persistent S. aureus mastitis in this dairy herd. PMID- 17917374 TI - Enhancement of hepatocellular proliferative activity of kojic acid in mice by a simultaneous administration of ascorbic acid. AB - To examine the tumor modification activity of kojic acid (KA) by sodium ascorbic acid (AA), 5-week-old male ICR mice were administered intraperitoneally with N diethylnitrosamine (DEN) as an initiation treatment. Two weeks after the initiation treatment, animals were fed basal diet containing 0 (Group 1: DEN alone) or 3% KA (Group 3: DEN+KA), drinking water containing 5,000 ppm AA (Group 2: DEN+AA) or 3% KA and 5,000 ppm AA (Group 4: DEN+KA+AA) for 6 weeks. One week after the administration of KA and/or AA, all mice were subjected to two-thirds partial hepatectomy. At the end of the experimental period, all surviving mice were sacrificed and removed the liver. The liver weights of the Groups 3 and 4 were significantly increased, and the number of proliferating cell nuclear antigen positive hepatocytes and the gene expressions of Ccnc, Ccnd1, Ercc and Cyp7a1 were significantly increased in the Group 4, as compared to the Group 1. These results of the present study suggest that AA enhances the hepatocellular proliferative activity of KA in mice. PMID- 17917375 TI - Relationship between plasma vitamin C and serum diagnostic biochemical markers in lactating cows. AB - We investigated the relationship between plasma vitamin C concentration and serum levels of some diagnostic biochemical markers in 118 lactating Holstein cows. Blood sample was collected once from each cow and we measured the plasma vitamin C concentration and the serum levels of glucose, beta-hydroxybutyrate, free fatty acids, triacylglycerol, total cholesterol, albumin, total bilirubin, alkaline phosphatase, aspartate aminotransferase and gamma-glutamyltransferase. The regression of plasma vitamin C with each serum diagnostic biochemical marker indicated that the vitamin C concentration significantly decreased as glucose, alkaline phosphatase or aspartate aminotransferase level increased and as total cholesterol or albumin concentration decreased. Furthermore, the plasma vitamin C concentration was significantly lower in the cows showing that each of these marker levels was out of its reference interval than in the cows showing that the marker level was within its reference interval. The significant correlations were observed among total cholesterol, albumin, alkaline phosphatase and aspartate aminotransferase levels, to which the glucose concentration was not related. These results showed that the plasma vitamin C concentration was low in the cows that had concurrently low levels of total cholesterol and albumin, and high levels of alkaline phosphatase and aspartate aminotransferase. Therefore, a hepatic malfunction possibly decreases plasma vitamin C concentration through suppressing vitamin C production. On the other hand, the high level of glucose possibly decreases plasma vitamin C concentration through suppressing vitamin C recycling. PMID- 17917376 TI - A retrospective study and gene analysis of canine sterile panniculitis. AB - In this study, a retrospective analysis was conducted to assess the current aspects and predisposing factors of canine sterile panniculitis. Miniature dachshund, neutered, and younger dogs appeared to be predisposed. In addition, histories of previous surgery and injection were associated in 46.5% of the cases, with several types of surgical suture materials used. About 88% of the dogs had multifocal lesions, frequently with signs of systemic illnesses. Whereas systemic immunosuppressive therapy was effective in most dogs, surgical excision of lesions was rarely curative. In order to prevent recurrences, over 65% of the cases required prolonged immunosuppressive therapy. Polymorphism of canine alpha(1)AT gene was investigated as a candidate gene for sterile panniculitis. Eight polymorphisms were discovered in seven Miniature dachshunds by direct nucleotide sequencing, which included a 12-bp deletion, three non-synonymous single nucleotide polymorphisms, and four silent substitutions. Genotyping of the two polymorphisms, c.109_120del12 and c.483A>C, which identified at high incidence in the dachshunds, was conducted in 83 dogs of 6 popular breeds. The frequencies of neither polymorphism differed between Miniature dachshunds and other breeds, suggesting that neither is responsible for developing panniculitis. PMID- 17917377 TI - Comparison of dendritic cell-mediated immune responses among canine malignant cells. AB - Dendritic cell (DC) vaccination is one of the most attractive immunotherapies for malignancies in dogs. To examine the differences in DC-mediated immune responses from different types of malignancies in dogs, we vaccinated dogs using autologous DCs pulsed with keyhole limpet hemocyanin (KLH) and cell lysate prepared from squamous cell carcinoma SCC2/88 (SCC-KLH-DC), histiocytic sarcoma CHS-5 (CHS-KLH DC), or B cell leukemia GL-1 (GL-KLH-DC) in vitro. In vivo inductions of immune responses against these tumor cells were compared by the delayed-type hypersensitivity (DTH) skin test. The DTH response against SCC2/88 cells were observed in dogs vaccinated with autologous SCC-KLH-DC, while the response was undetectable against CHS-5 and GL-1 cells in dogs vaccinated with autologous CHS KLH-DC and GL-KLH-DC. Skin biopsies taken from DTH challenge sites were then examined for immunohistochemistry, and recruitment of CD8 and CD4 T cells was detected at the site where SCC2/88 cells were inoculated in dogs vaccinated with SCC-KLH-DC. By contrast, neither CD8 nor CD4 T cell infiltration was found at the DTH challenge site in the dogs vaccinated with CHS-KLH-DC or GL-KLH-DC. These findings may reflect that the efficacy of immune induction by DC vaccination varies among tumor types and that immune responses could be inducible in squamous cell carcinoma. Our results encouraged further investigation of therapeutic vaccination for dogs with advanced squamous cell carcinoma in clinical trials. PMID- 17917378 TI - The A y allele at the agouti locus enhances sensitivity to endotoxin-induced lethality in mice. AB - In the course of investigations on anorexia during infection, I found that B6 A(y) mice had significantly increased sensitivity to lipopolysaccharide (LPS) induced lethality as compared with isogenic B6 mice. I also found that the sensitivity to the lethal effect of LPS dramatically increased in aged mice (age effect), both B6 and B6-A(y). However, the A(y) effect of enhancing sensitivity to LPS-induced lethality was still significant, suggesting that the A(y) effect is independent of age. In the absence of tumor necrosis factor alpha (TNFalpha), the A(y) effect was still significant, suggesting that the A(y) effect is independent of TNFalpha toxicity. A dose of LPS of 100 mug per mouse caused 15% lethality in B6, 65% in B6-A(y) (significantly higher than B6), and 100 % in leptin-deficient B6-ob/ob (significantly higher than B6 and B6-A(y)). The results support the hypothesis that endogenous leptin has a protective role against infection, and that a part of this leptin effect is mediated by alpha-melanocyte stimulating hormone (alphaMSH). In contrast to the results of simple blockade at the melanocortin 4 receptor (MC4R), B6-A(y) suffered more severe LPS-induced anorexia than did B6; therefore, the pathway involving MC4R is not absolutely required for the LPS-induced anorexia, and the presence of pathways involving other melanocortin receptor types was suggested. Because alphaMSH is suggested to be an endogenous anti-inflammatory peptide, and because melanocortin 1 receptor (MC1R) is expressed in various cutaneous cell types, the A(y) effect might be caused via the pathway involving MC1R. Physiologic significance of alphaMSH-MC1R interaction in host defense against infection is discussed. PMID- 17917379 TI - Influence of preemptive analgesia with meloxicam before resection of the unilateral mammary gland on postoperative cardiovascular parameters in dogs. AB - Preemptive analgesia is recommended in small animal medicine. However, many studies have evaluated the response to analgesic treatment by behavioral observation. Therefore, the influence of preemptive analgesia with meloxicam on postoperative cardiovascular and renal parameters remains to be clarified. The present study examined the changes in blood pressure, heart rate, double product and heart rate variability for 14 days and the changes in glomerular filtration rate (GFR) and serum cortisol level for 24 hr after resection of the unilateral mammary gland in meloxicam and control groups consisting of 5 healthy dogs. All data were collected under unanesthetized and unrestrained conditions using a radio telemetry system. Blood pressure, heart rate and double product were significantly lower in the meloxicam compared with the control group, and the meloxicam group's diurnal changes became stable more than 36 hr earlier than those of the control group. The systolic, diastolic and mean blood pressure values of the meloxicam group were 5- to 20-mm Hg lower than those of the control group until 5 days after surgery. The maximum difference between the two groups in terms of the double product values 14 days after surgery was 2,000 bpm x mmHg. Autonomic activity inhibition was prolonged in the control group. There were no significant differences in the 24-hr changes in GFR or serum cortisol level. This study showed that perioperative administration of meloxicam reduced unfavorable postoperative changes in the cardiovascular system without influencing renal function. PMID- 17917380 TI - Assessing equine anxiety-related parameters using an isolation test in combination with a questionnaire survey. AB - The aim of the study was to determine parameters reflecting equine anxiety trait by comparing results obtained in a behavior test and an anxiety score assessed by familiar caretakers in response to a questionnaire. In the behavior test, horses were individually led into a novel room by their caretakers and loosely tethered to decrease excessive movement using the common cross-tying technique with two leads and breakable plastic cords. The horses initially remained with their caretaker for 2 min; the caretaker then left and the subject animal was left alone for 2 min. The latency to break the plastic cords, heart rate, the number of steps and pawings were recorded. When the horses were left alone, most parameters changed significantly and some showed sexual differences. A correlation analysis revealed that anxiety score assessed by caretakers showed a negative correlation with the latency to break the plastic cord and a positive correlation with heart rate only when horses were isolated. These two were suggested feasible parameters for assessing the anxiety trait of unfamiliar horses. Both the behavioral results and the anxiety scores also indicated that females were more anxious than males. Our results suggest that it would be a useful strategy for assessing other temperament traits as well to combine behavior tests with a questionnaire survey. PMID- 17917381 TI - Therapeutic trial of granulocyte-colony stimulating factor for dilated cardiomyopathy in three dogs. AB - Three dogs were presented to us for evaluation of cardiac problems. Electrocardiographic recordings revealed severe tachyarrhythmia and atrial fibrillation with ventricular tachycardia in 2 of the 3 dogs. The echocardiographic findings of the 3 dogs revealed markedly decreased fractional shortening and a marked increase in E-point septal separation. Based on the results of electrocardiographic and echocardiographic evaluation, the 3 dogs were diagnosed as dilated cardiomyopathy (DCM). The dogs were treated with conventional cardiac medication, but cardiac function did not improve and the clinical signs remained. We subsequently attempted treatment with granulocyte colony stimulating factor (G-CSF; 10 microg/kg, subcutaneously). The specific purpose of G-CSF therapy for DCM was to improve cardiac function and a significant improvement in cardiac function was confirmed. The three dogs had no treatment side effects. This case report suggests that G-CSF might have therapeutic effects for medically refractory DCM in dogs. PMID- 17917382 TI - Fatal winter dysentery with severe anemia in an adult cow. AB - An adult dairy cow fatally affected with winter dysentery was investigated pathologically and virologically. The cow had severe anemia and diarrhea with massive blood. Pathologically, the loss of surface epithelial cells and necrosis of crypt epithelial cells in the large intestine were observed. Bovine coronavirus (BCV) antigen was observed in necrotic crypt epithelial cells of the large intestine. Virus particles were found in the necrotic epithelial cells of the large intestine. Virologically, BCV was isolated from the feces of the dead cow. The dead cow had no serum antibody against BCV although the co-habitants did. These suggest that severe infection of BCV in the cow without the BCV antibody accompanied by severe hemorrhagic anemia resulted in the cow's death. PMID- 17917383 TI - Isolation of Actinobacillus pleuropneumoniae serovar 15-like strain from a field case of porcine pleuropneumonia in Japan. AB - An Actinobacillus pleuropneumoniae strain isolated from a field case of porcine pleuropneumonia in Japan, was closely related to a reference strain of serovar 15, which is a newly proposed serovar according to an analysis of field isolates originating from Australia. The isolate had biological and biochemical properties consistent with A. pleuropneumoniae biovar 1, and reacted strongly to a rabbit antiserum raised against a reference strain of serovar 15 in an agar gel precipitation test. The nucleotide sequence of a hyper variable region in the 16S RNA gene of the isolate was identical to that of the reference strain of serovar 15. The isolate possessed A. pleuropneumoniae-RTX toxin (Apx) II, III, and IV genes, consistent with serovar 15. Its virulence in mice was lower than that of ApxI-bearing strains but higher than that of other ApxIII-bearing strains. This is the first report describing the isolation of A. pleuropneumoniae serovar 15 like strain from a country or region other than Australia. PMID- 17917385 TI - Streptozotocin-induced diabetes increases apoptosis through JNK phosphorylation and Bax activation in rat testes. AB - Diabetic disease is known to suppress male reproductive activity in laboratory animals and humans. The present study was designed to evaluate whether streptozotocin-induced diabetes increases apoptotic cell death in rat testes through activation of the JNK and Bax pathway. Diabetes was induced by a single intravenous injection of streptozotocin (40 mg/kg) and testis samples were collected after 3 months. Compared with controls, body weight and testicular weight were lower in the diabetic group, and the apoptotic index in testicular germ cells was significantly increased. Expression of phospho-JNK and Bax was significantly increased in the diabetic group, and the level of activated caspase 3 was also increased, compared to that of controls. Our findings suggest that streptozotocin-induced diabetes increases apoptotic cell death in rat testes through phosphorylation of JNK and activation of Bax. PMID- 17917384 TI - Clinical evaluation of left ventricular moderator band in 12 dogs. AB - We retrospectively examined clinical data for 12 dogs in which echocardiography revealed the presence of left ventricular moderator bands (LMB). Physical examinations, electrocardiography and echocardiography revealed slight cardiac murmurs, increasing QRS complex and left ventricular turbulent flow (6 of the dogs), respectively. No differences were observed with respect to gender, and no specific clinical symptoms or types of dog that frequently develop this disorder were found. PMID- 17917386 TI - Colitis associated with Clostridium difficile in specific-pathogen-free C3H-scid mice. AB - Soft feces and a decreased delivery rate were observed in a specific-pathogen free (SPF) C3H-scid mouse breeding colony. Grossly, the ceca were shrunken and edematous in the affected mice. Histopathologically, severe edema in the cecal submucosa as well as infiltration of inflammatory cells in the lamina propria and submucosa of the ceca and colon were observed. No pathogenic microorganisms were detected by the routine microbiological tests. By anaerobic bacterial examination, Clostridium (C.) difficile with toxin A was isolated from the cecal contents of the affected mice. The mice were diagnosed with C. difficile associated colitis. This case appears to be the first report of natural infection with C. difficile in SPF mice with clinical signs. PMID- 17917388 TI - A case of adenocarcinoma of the endometrium extending into the leiomyoma of the uterus in a rabbit. AB - In a pet rabbit, 2 tumor masses one on each horn were macroscopically seen in the wall of the uterus. On light microscopic examination, the right horn mass consisted of an admixture of neoplastic epithelial and mesenchymal element. The epithelial element was composed of neoplastic epithelial cells with numerous mitotic figures and formed varied sizes of acini, glandular, and solid structures. The tumor was diagnosed as an adenocarcinoma of the endometrium. The mesenchymal element was composed of well-differentiated smooth muscle cells and was diagnosed as a leiomyoma. While adenocarcinoma cells formed a protrusive mass in the uterine lumen, they also showed an extension into the leiomyoma of the myometrium. By immunohistochemistry, adenocarcinoma stained positive for cytokeratin (MNF116) and leiomyoma stained positive for smooth muscle actin, showing a substantial difference in the cytological nature of these tumor cells. The results may give a further evidence supporting the narrative of the tumor development that an adenocarcinoma of the endometrium extended into leiomyoma of the uterus. To the author's knowledge, this is the first report describing this type of combination of two independent tumors in a pet rabbit. PMID- 17917387 TI - A case of atypical canine lymphoma with oral mass and multiple osteolysis. AB - An 8-year-old female Golden Retriever had an oral mass and lameness. Multiple osteolysis of the systemic skeleton without monoclonal gammopathy was shown on electrophoresis of serum and urine samples. Cytological and histopathological examinations of the oral mass revealed atypical polymorphic cells similar to myeloid cells, and bone marrow aspiration indicated that these abnormal cells also might have invaded the bone marrow. These cells were negative to peroxidase and non-specific esterase staining, and clonal expansion of B lymphocytes could be detected by polymerase chain reaction (PCR) assay for antigen receptor gene rearrangement. The case was diagnosed as atypical lymphoma and treated by multi drug chemotherapy. On the 142nd day after the first admission, the case had remission and the oral mass and multiple osteolysis were improved. PMID- 17917389 TI - Propionate tolerance test for determination of insulin secretion in a hyperglycemic Japanese Black steer. AB - A propionate tolerance test (PTT) was used to determine the pathophysiology of a Japanese Black steer with hyperglycemia. In the hyperglycemic steer, a low insulin secretion was confirmed by a glucose tolerance test (GTT), so that the hyperglycemic steer was diagnosed as insulin-dependent diabetes mellitus. Although the plasma insulin concentration in the control cattle increased in response to propionate stimulation, a low insulin response to PTT was observed in the diabetic steer. The fact that both PTT and GTT determined that the diabetic steer had low insulin secretion suggests that the PTT might be an effective diagnostic tool for diabetes mellitus in cattle. PMID- 17917390 TI - Establishment of a novel equine cell line for isolation and propagation of equine herpesviruses. AB - In the present study, an equine-derived cell line was established by transfecting primary fetal horse kidney (FHK) cells with expression plasmid encoding simian virus 40 (SV40) large T antigen and then cloning them by limiting dilution. The cloned cell line, named FHK-Tcl3, grew well and could be propagated over 30 times by splitting them 1:3. Equine herpesvirus (EHV)-1 and EHV-4 replicated well in FHK-Tcl3. EHV-2 and EHV-4 were isolated from samples collected from horses in the field using FHK-Tcl3, and EHV-3 also propagated in FHK-Tcl3. These results indicated that this novel cell line, FHK-Tcl3, can be used for isolation and propagation of equine herpesviruses. PMID- 17917391 TI - [Indications for antiviral therapy for chronic hepatitis B]. PMID- 17917393 TI - [Therapy for chronic hepatitis B: relationship between HBV genotype and efficacy or resistance of anti-viral therapy]. PMID- 17917392 TI - [Natural course and recommendation of therapy for HBV infection]. PMID- 17917394 TI - [Recent therapies for hepatitis B (discussion)]. PMID- 17917395 TI - [Case of ileus due to an appendix epiploica]. AB - A 79-year-old woman who had an abdominal pain and vomiting admitted to the hospital with a diagnosis of ileus. An ileus tube was inserted and the fluoroscopic study of the small intestine revealed narrowing of the two parts, jejunum and the ileum. Because no improvement was obtained by conservative treatment, an operation was performed. We observed that an appendix epiploica of the sigmoid colon extended long, and its tip was adherent to the retroperitoneum. The small intestine was impacted into the aperture formed by the band. Seventeen cases with intestinal obstruction due to an appendix epiploica have been reported in Japan, including this case. PMID- 17917397 TI - [Carcinoma arising from ileorectal fistula in a patient with Crohn's disease]. AB - A 72-year-old man was given a diagnosis of Crohn's disease in 1976 at age 44, and partial resection of is ileum was performed. In November 1982, barium enema examination revealed on ileorectal fistula. As he had no complaint, conservative therapy was chosen. In August 2003, he had high fever and CT scan revealed presacral abscess. Ileocecal resection, partial resection of is small intestine and loop sigmoid colostomy were performed. In December 2004, the serum level of CEA was gradually elevated and he complained of anal mucus discharge. Endoscopic examination showed a fistula orifice in the rectum and biopsy of the fistula revealed mucinous adenocarcinoma. We performed abdominoperineal resection of the rectum with partial resection of the sacrum. We thought that careful observation helped the detection of such a rare case of carcinoma arising from a fistula tract. PMID- 17917396 TI - [Elderly case of systemic lupus erythematosus who showed lupus peritonitis as the initial symptom]. AB - A 64-year-old woman who admitted because of abdominal pain and ascites. She was diagnosed as having systemic lupus erythematosus (SLE) and lupus peritonitis based on anti ds-DNA antibody, ANA positive, serositis and renal dysfunction. She was successfully treated by intravenous pulse therapy with methylprednisolone, but her enteritis recurred after switching to oral administration. She recovered following pulse therapy with methylprednisolone and extracorporeal apheresis. This case was characterized by a rather old age of onset and the peculiar initial symptoms of peritonitis. PMID- 17917398 TI - [Case of ruptured gastroduodenal arterial pseudoaneurysm with penetration to the duodenum, successfully treated with transcatheter arterial embolization]. AB - A 77-year-old man, who underwent segmental pancreatectomy for intraductal papillary mucinous adenoma in 2001, was referred to our hospital with complaints of hematemesis and melena on January, 2004. Emergency upper gastrointestinal endoscopy showed a pulsating submucosal protrusion in the duodenal bulb, which was identified as a gastroduodenal arterial aneurysm measuring 1.5cm on abdominal CT imaging. Transcatheter arterial embolization of the aneurysm with metallic coils was successfully performed. Periodically repeated endoscopic examination has revealed the coils protruding into the duodenal lumen without any serious complication. PMID- 17917399 TI - [Congenital absence of the portal vein in an adult woman with liver tumor]. AB - A 30-years-old Japanese woman with a liver tumor was found to have congenital absence of the portal vein (CAPV). Both three-dimensional CT and angiography revealed that the superior mesenteric vein and splenic vein flowed into inferior vena cava and there was us portal vein, CAPV is an extremely rare congenital anomaly and liver tumor. Most cases on diagnosed in childhood, although this case was found in on adult. We reviewed the literature on reported CAPV cases. PMID- 17917400 TI - [Pyogenic liver abscess complicated by gastric fistula and bacterial meningitis]. AB - A 77-year-old woman was admitted suffering from fever and headache. On laboratory examination, bacterial meningitis and sepsis due to Klebsiella pneumoniae were diagnosed. In addition, a hepatic cystic lesion measuring 13 cm in diameter in the left lobe was indicated on diagnostic imaging. After treatment with antibiotics, her signs of infection improved and the hepatic lesion decreased in size. After discharge, however, the cystic liver mass increased and a gastric fistula developed. Hepatic and gastric resections were performed because of the possibility of biliary cystadenocarcinoma and gastric invasion. Pathologically, a pyogenic liver abscess complicated by gastric fistula was diagnosed. PMID- 17917401 TI - [Two cases of pyogenic spondylitis with chronic hepatitis C during combination therapy of interferon alfa and ribavirin]. AB - This report describes our experience with two cases of pyogenic spondylitis with chronic hepatitis C during combination therapy of interferon alfa and ribavirin. The first patient, a 59-year-old man, was treated conservatively and improved, but the second patient, a 69-year-old woman, was not improved by conservative therapy and reconstructive operation was performed. The combination therapy of interferon alfa and ribavirin has a high risk of severe infectious diseases as side effects. CT scan and MRI are recommended immediately to diagnose pyogenic spondylitis, when patients has pyrexia and lumbago with laboratory data suspected inflammation during interferon therapy. PMID- 17917402 TI - [Case of pancreatic endocrine tumor associated with Cushing's syndrome]. AB - A 78-year-old woman was admitted complaining of edema of the bilateral lower extremities and face. Computed tomography (CT) and ultrasonography (US) of her abdomen revealed a pancreatic tumor and multiple liver metastases. After admission, hypokalemia and muscle weakness and edema of the bilateral lower extremities rapidly worsened. The diagnosis of Cushing's syndrome was established based on clinical and biochemical data and endocrine studies. We thought that the primary tumor was a pancreatic endocrine tumor based on the liver tumor biopsy findings, and that the pancreatic tumor and liver metastatic tumors were ectopic ACTH-producing tumors. A case of pancreatic endocrine tumor associated with Cushing's syndrome is relatively rare. We summarize previous reports. PMID- 17917403 TI - Simple and easy assessment of falling risk in the elderly by functional reach test using elastic stick. AB - Dynamic balance ability related to maintaining postural stability during movement is closely tied to fall risk in the elderly. The functional reach (FR) test has been developed to evaluate their dynamic balance. Although a simple and new FR test using an elastic stick has been proposed by modifying the above original FR test, the abilities related to both FR tests are judged to differ because of the large difference in the testing method. This study aimed to compare center of gravity fluctuation, muscle activity and functional reach distance as measured by the original FR test and the elastic stick FR test. First, reach distance, back/forth and right/left moving distance of the center of gravity, and activity of the lower leg muscles (soleus and tibialis anterior) were compared between both tests based on data obtained from 30 young male adults. All parameters except for the right/left moving distance were significantly larger in the elastic stick FR test. Next, the reach distance was examined in both FR tests using 53 elderly subjects; it was significantly longer in the elastic stick FR test, but showed no significant sex difference. The reach distance in both tests was significantly shorter (about 7 cm) in the elderly than in young adults. In conclusion, the elastic stick FR test involves greater leg muscle strength exertion and forward transferring of the center of gravity as compared with the original FR test. Because the elastic stick FR test relates largely to leg muscle function and equilibrium function, it may be more useful for evaluating the dynamic balance ability of the elderly. PMID- 17917404 TI - Intracoronary stem cell infusion in heart transplant candidates. AB - The stem cell transplantation is emerging as a potential therapeutic modality for patients with heart failure. It has been demonstrated that intracoronary stem cell transplantation had beneficial effects on left ventricular perfusion and contractile functions. We hypothesized that patients with end-stage ischemic cardiomyopathy, who are candidates for heart transplantation, could also benefit from autologous intracoronary stem cell transplantation. We performed a prospective, open-labeled study in 10 patients with end-stage ischemic cardiomyopathy, who were on the waiting list for heart transplantation. Each patient received bone marrow-derived mononuclear cell infusion via balloon catheter in the target vessel, which had been revascularized by percutaneous intervention and was patent before the procedure. Clinical and laboratory evaluations, a treadmill exercise test, echocardiography, and single photon emission tomography (SPECT) were performed to the patients at baseline and 6 months after stem cell infusion. At 6-month follow-up of the eight patients who were able to complete the study, we revealed a significant increase in ejection fraction (from 30.0 +/- 6.6% to 36.2 +/- 7.3%; p = 0.001) in echocardiographic evaluation. SPECT evaluation also displayed a reduction in infarct area (50.4 +/- 16.1% to 44.1 +/- 12.5%; p = 0.003). Both myocardial oxygen consumption (p = 0.001) and metabolic equivalents (p = 0.001) were significantly increased at 6 month follow-up. These results demonstrate that intracoronary stem cell transplantation ameliorates heart failure symptoms and improves left ventricular function and perfusion. Therefore intracoronary stem cell transplantation may be used as an alternative treatment option for heart transplant candidates. PMID- 17917405 TI - Identifying medical interview behaviors that best elicit information from patients in clinical practice. AB - Many textbooks indicate the desirability of using specific medical interview behaviors to obtain information from patients, although little evidence has been presented. The aim of this study is to investigate the relationships between the use of medical interview behaviors and the amount of information obtained. The behaviors studied included: open-ended questions, facilitation (comments or interviewer behavior that encourages the patient to continue talking), the open to-closed cone (the gradual narrowing of focus from an initial nondirective approach to a more direct exploration), summarization (providing the patient with an explicit verbal summary of the information gathered thus far), and surveying problems. The subjects were 315 patients who visited an outpatient facility. These medical interviews were videotaped and reviewed by a trained rater using the Takemura Medical Interview Rating Scale, which was developed to assess the use of particular medical interview behaviors and to measure the amount of information obtained from patients regarding their chief physical complaints. Significant positive relationships were found between three particular interview behaviors and the amount of information obtained: facilitation, the open-to closed cone, and summarization. These positive relationships were still present after adjusting for other medical interview behaviors used, and after adjusting for the time duration of the medical interviews (F = 15.3, p < 0.0001; F = 40.1, p < 0.0001; F = 5.57, p = 0.019, respectedly). This study reveals a positive relationship between three specific medical interview behaviors (facilitation, the open-to-closed cone, and summarization) and the amount of information obtained in a real clinical practice setting. PMID- 17917406 TI - Increased phospholipid transfer protein activity associated with the impaired cellular cholesterol efflux in type 2 diabetic subjects with coronary artery disease. AB - Reverse cholesterol transport (RCT) is the pathway, by which the excess of cholesterol is removed from peripheral cells to the liver. An early step of RCT is the efflux of free cholesterol from cell membranes that is mediated by high density lipoproteins (HDL). Phospholipid transfer protein (PLTP) transfers phospholipids between apolipoprotein-B-containing lipoproteins (i.e., chylomicrons and very low-density lipoproteins) and HDL. PLTP contributes to the HDL maturation and increases the ability of HDL to extract the cellular cholesterol. It is known that RCT is impaired in type 2 diabetic patients, especially when cardiovascular complication is associated with. In this study, we measured the serum capacity that promotes cellular cholesterol efflux and the plasma PLTP activity in type 2 diabetic patients with coronary artery disease (CAD) (n = 35), those without CAD (n = 24), and 35 healthy subjects as a sex- and age-matched control. In patients with CAD, plasma triglyceride level was higher compared to controls (p < 0.01) and HDL-cholesterol was lower (p < 0.01 vs control and the patients without CAD). In diabetic patients with or without CAD, PLTP activity was consistently increased, compared to controls, while cellular cholesterol efflux activity was decreased by 20% (p < 0.001) or 13.5% (p < 0.01), respectively. In conclusion, plasma PLTP activity was increased in type 2 diabetic patients with or without CAD, which could impair cellular cholesterol removal and might accelerate atherosclerosis in diabetic patients. PMID- 17917407 TI - Colchicine, a microtubule depolymerizing agent, inhibits myocardial apoptosis in rats. AB - Heart failure is the most common cardiovascular disease with high mortality and morbidity. Both enhanced microtubule polymerization and cardiomyocyte apoptosis are involved in the pathogenesis of heart failure. However, the link between the two mechanisms remains to be elucidated. In this study, we thus address this important issue in cultured cardiomyocytes from Wistar rats in vitro and in angiotensin II (ATII)-infused rats in vivo. Confocal microscopy examination showed that in cultured rat cardiomyocytes, micrographic density of microtubules was increased by paclitaxel, a microtubule-polymerizing agent, and decreased by colchicine, a microtubule-depolymerizing agent, but not affected by ATII, isoproterenol, or tumor necrosis factor-alpha alone. Immunoblotting analysis showed that Bax/Bcl-2 ratio, which is associated with the activation of caspase 3, was significantly increased in ATII-stimulated cultured cardiomyocytes in vitro and in ATII-infused rats in vivo, both of which were inhibited by co treatment with colchicine. Caspase-3 and TUNEL assay to detect apoptosis in vitro demonstrated that paclitaxel or ATII alone significantly enhanced and their combination further accelerated cardiomyocyte apoptosis, which was again significantly inhibited by colchicine. Caspase-3 and TUNEL assay in vivo also demonstrated that ATII infusion significantly increased myocardial apoptosis and that co-treatment with colchicine significantly suppressed the apoptosis. In conclusion, these results indicate that a microtubule-depolymerizing agent could be a potential therapeutic strategy for treatment of heart failure. PMID- 17917408 TI - Dosimetric factors used for thoracic X-ray radiotherapy are not predictive of the occurrence of radiation pneumonitis after carbon-ion radiotherapy. AB - Radiation pneumonitis (RP) is one of the most common dose-limiting toxicities in thoracic X-ray radiotherapy (XRT). Dosimetric factors are used for prediction of the occurrence of RP after XRT. Carbon-ion radiotherapy (CRT) is a promising modality because of its excellent dose localization and high biological effect on tumors. This study aims to analyze the relationship between dosimetric factors developed for XRT and the incidence of RP in patients with stage I non-small cell lung cancer (NSCLC) after CRT. We examined 80 inoperable patients with NSCLC. The ranges of the daily fraction sizes and the total doses were from 3.3 to 8.8 GyE and from 59.4 to 95.4 GyE, respectively. These doses were successfully delivered with acceptable toxicity; >or= grade 2 RP was observed in 8 patients (10%). The severity of RP was graded within 6 months of the initiation of CRT using the Radiation Therapy Oncology Group criteria. These results indicate the excellent dose distribution of CRT. We then compared the dosimetric data of the 8 patients developed >or= grade 2 RP with those of 72 patients developed 5, > 20, and > 30 GyE, and mean lung dose, were not predictive factors for RP after CRT. The dosimetric factors used for XRT are not applicable for CRT in patients with NSCLC. The dosimetric factors for CRT remain to be developed. PMID- 17917409 TI - Long-term beneficial effects of lung volume reduction surgery on quality of life in patients with chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is characterized by progressive airflow limitation, which results in exertional dyspnea and physical disability. Subsequently, those cause a difficulty in performing routine activities of daily living and affect their health-related quality of life (HRQOL). Lung volume reduction surgery (LVRS) has been reported to be an effective treatment for selected patients with advanced COPD to improve pulmonary function, lung mechanics, exercise tolerance, and dyspnea. However, the long-term effects of LVRS on HRQOL have not been fully investigated. Therefore the effects of LVRS on generic and disease-specific HRQOL were assessed in patients with COPD following LVRS for 36 months. Nineteen patients (65.1 +/- 7.0 [mean +/- S.D.] years old) who underwent pulmonary rehabilitation plus LVRS (LVRS group), and 8 patients (67.2 +/- 5.8 years old) who did pulmonary rehabilitation but not LVRS (Medical group) were studied. In both groups, optimal medication was given throughout this period. Generic HRQOL and disease-specific HRQOL were evaluated before rehabilitation, and 3, 12, 24, and 36 months after LVRS. Following LVRS, the generic HRQOL was significantly improved and the disease-specific HRQOL was maintained up to 36 months. In Medical group, disease-specific HRQOL rapidly deteriorated. In conclusion, the long-term effects of LVRS on HRQOL in COPD patients were maintained up to 36 months compared with Medical group. Both generic and disease-specific HRQOL changed differently, suggesting the importance of both assessments especially in long-term follow up. PMID- 17917410 TI - Increased rates of intense nasopharyngeal bacterial colonization of Vietnamese children with radiological pneumonia. AB - Acute lower respiratory infection (ALRI), primarily pneumonia, is the leading cause of death in children under the age of five. Bacterial ALRI is preceded by asymptomatic bacterial colonization. Bacterial colonization, therefore, may have an important role in the development of pneumonia in children. This case-control study was conducted in order to determine if intense bacterial colonization was increased in the nasopharynx of pediatric patients with ALRI. One hundred-sixty four pediatric patients with ALRI and 70 healthy children < 5 years of age were enrolled in Hanoi, Vietnam between 2001 and 2002. Bacterial pathogens were isolated from nasopharyngeal secretions and quantitatively cultured. Of 164 patients, 91 were diagnosed as having radiological pneumonia (PN group) and 73 as having acute bronchitis (AB group). Intense growth of any bacterial pathogen (>or= 10(6) colony-forming units/ml) was highest in the PN group (49.4%), followed by the AB group (28.8%), with healthy children having the lowest (17.1%). Patients with intense bacterial growth were more likely to develop pneumonia, but not acute bronchitis, than were patients with light or no bacterial growth. The results of this case-control study suggest that the vertical spread of intense bacterial pathogens colonized in the nasopharynx to the lower airway leads to bacterial pneumonia in children under the age of five. PMID- 17917411 TI - Preoperative clinical status but not waiting time predicts in-hospital outcomes of surgery in patients with left main coronary artery stenosis. AB - Contoversy exists about the optimal operation time of the patients with left main coronary artery (LMCA) stenosis. We therefore, aimed to investigate the effect of waiting time on in-hospital morbidity and mortality in patients with LMCA stenosis and identify the risk factors associated with adverse cardiovascular events before and during surgery. One hundred seventy six patients with LMCA stenosis were divided into two groups according to the time period between coronary angiography and coronary artery bypass surgery (group 1: 7 days, 82 patients). Primary end points were death and major adverse cardiac event (MACE): in-hospital death, sustained ventricular tachycardia or ventricular fibrillation development, postoperative stay in the intensive care > 48 hrs and in hospital > 9 days. Demographic and clinical characteristics of patients in groups 1 and 2 were comparable. There was no difference between the two groups in terms of in-hospital morbidity, mortality and MACE. When we analyzed the differences between the patients with and without MACE, the patients who experienced MACE were older (p = 0.001), and had higher degree of LMCA stenosis (p = 0.01), higher degree of right coronary artery stenosis (p = 0.02), higher blood urea level (p = 0.003), and higher incidence of unstable angina or myocardial infarction within 2 weeks (p = 0.001). Independent risk factors for MACE were unstable angina or myocardial infarction within 2 weeks, age more than 70 years and stenosis more than 75% in the LMCA. These results suggest that preoperative clinical status but not waiting time predicts in-hospital surgical outcomes in LMCA stenosis. PMID- 17917412 TI - The high incidence of the emergence of entecavir-resistant mutants among patients infected with lamivudine-resistant hepatitis B virus. AB - Hepatitis B virus (HBV) infection remains to be one of the most prevailing infection in the world, causing chronic liver diseases. Although lamivudine has been effective to suppress HBV replication, longer durations of administration can lead to the emergence of drug-resistant mutant viruses, followed by reactivation of hepatic inflammation (breakthrough hepatitis). Moreover, the optimal period of administration as well as the effects of anti-viral nucleot(s)ide such as lamivudine, adefovir, and entecavir, has not been established. To evaluate the efficacy of the anti-viral effects of entecavir for lamivudine-resistant HBV, we administered entecavir sequentially in four patients with chronic HBV infection, who demonstrated the emergence of lamivudine resistant HBV and histological active hepatitis. The antiviral effects were evaluated by the serum viral loads and biochemical laboratory data. After follow up periods of more than 36 months, we found high incidence in the emergence of entecavir resistant mutants (3/4, i.e., 75%). An additional mutation at the 184th amino acid, different from the previously reported lamivudine-resistant mutations (80th, 180th, and 204th), seemed to have a close relationship with the induction of entecavir-resistant mutants at least for Japanese HBV genotype C. Our observation draws attention to the possibility that the usage of entecavir for lamivudine-resistant HBV could promptly induce entecavir-resistant mutations in addition to lamivudine-resistance. PMID- 17917413 TI - [Novel material and strategy for gene delivery]. PMID- 17917414 TI - [In vivo siRNA delivery to tumor cells and its application to cancer gene therapy]. AB - RNA interference (RNAi) is a posttranscriptional gene-silencing event in which short double-stranded RNA (siRNA) degrades target mRNA. Because of its potent and highly specific gene-silencing effect, RNAi is expected to be used in the treatment of various diseases. Cancer is one of the major targets of RNAi-based therapy, because silencing oncogenes or other genes contributing to tumor progression can be target genes for RNAi. The delivery of RNAi effector to target cells is one of the key factors determining therapeutic efficacy, because gene silencing is limited to cells reached by RNAi effectors. Tumor cell lines stably expressing reporter genes were confirmed to be effective in sensitively and quantitatively evaluating RNAi effects in tumor cells in vitro and in vivo. Quantitative analyses of the gene-silencing effect revealed that short-hairpin RNA expressing plasmid DNA (pshRNA) has more durable effects than siRNA. Intratumoral injection of RNAi effectors was effective in suppressing target gene expression in tumor cells, and silencing of beta-catenin or hypoxia-inducible factor-1alpha (HIF-1alpha) significantly inhibited tumor growth. RNAi effectors were successfully delivered to tumor cells colonizing the liver through the vascular route. We found that tumor-bearing liver showed elevated HIF-1alpha expression in the cells, and the silencing of the expression in normal liver cells is also effective in inhibiting metastatic tumor growth. These results indicate the possibility of RNAi-based cancer therapy. PMID- 17917415 TI - [Immunotherapy against peritoneal dissemination by immunostimulatory CpG DNA]. AB - Peritoneal dissemination is one of the most common causes of metastasis from malignancies in the abdominal cavity. However, the treatment of peritoneal dissemination is difficult; patients receiving normal chemotherapy have a 0-1% chance of surviving for 5 years. Milky spots in the greater omentum are considered to facilitate the adhesion and invasion of abdominal free cancer cells, and subsequently lymph node metastasis occurs. Since immune cells such as macrophages and lymphocytes are present in the greater omentum and lymph nodes, the activation of immune cells would be a promising strategy for treatment. Single-stranded oligonucleotides containing CpG dinucleotides (CpG DNA) are recognized by Toll-like receptor-9 on antigen-presenting cells such as macrophages to stimulate Th-1-type immune responses. However, a delivery system for CpG DNA to immune cells is essential to develop effective therapy against peritoneal dissemination. Here we review the pathophysiologic basis of peritoneal dissemination and introduce our approach that employs cationic liposomes as a carrier for CpG DNA as a new approach in the treatment of peritoneal dissemination. PMID- 17917416 TI - [Development of gene delivery system using PLGA nanospheres]. AB - The development of nonviral vectors for the efficient and safe delivery to cells has long been awaited to facilitate gene therapy. Recently, many nonviral vectors modified with cationic lipids, cationic polymers, etc. have been reported. However, those nonviral vectors with cationic materials require improved stability, longer duration of gene expression, and reduced cytotoxicity. We successfully prepared mucoadhesive poly (lactide-co-glycolide) nanospheres (PLGA NS) by modifying the nanoparticulate surface with chitosan to improve mucosal peptide absorption after oral and pulmonary administration. Furthermore, we found that nucleic acid, which was not dispersed in the organic solvent, could be dispersed by forming a complex with cationic lipid. Using this phenomenon, polynucleic acids for gene therapy (plasmid DNA, antisense oligonucleotide, small interfering RNA, etc.) can be encapsulated into the matrix of the polymer particles with the emulsion solvent diffusion method. The advantages of this preparation method are its simple process and avoidance of an ultrasonication process for submicronization of particles. The resultant nanospheres show better cellular uptake and different gene therapeutic effects compared with conventional vectors due to their improved adherence to cells and sustained release of polynucleic acid in the cells. In conclusion, chitosan-coated PLGA NS can possibly be applied in nonviral vectors for gene therapy. PMID- 17917417 TI - [Development of a novel systemic gene delivery system for cancer therapy with a tumor-specific cleavable PEG-lipid]. AB - For successful cancer gene therapy via intravenous administration, it is essential to optimize the stability of carriers in the systemic circulation and the cellular association after the accumulation of the carrier in tumor tissue. However, a dilemma exists regarding the use of poly(ethylene glycol) (PEG), which is useful for conferring stability in the systemic circulation, but is undesirable for the cellular uptake and subsequent processes. We report the development of a PEG-peptide-lipid ternary conjugate (PPD). In this strategy, PEG is removed from the carriers via cleavage by a matrix metalloproteinase (MMP), which is specifically expressed in tumor tissues. An in vitro study revealed that the PPD-modified gene carrier (multifunctional envelope-type nano device, MEND) exhibited pDNA expression activity that was dependent on the MMP expression level in the host cells. In vivo studies further revealed that the PPD was potent in stabilizing MEND in the systemic circulation and facilitating tumor accumulation. Moreover, the intravenous administration of PPD or PEG/PPD dually modified MEND resulted in the stimulation of pDNA expression in tumor tissue, as compared with a conventional PEG-modified MEND. Thus MEND modified with PPD is a promising device with the potential to make in vivo cancer gene therapy achievable. PMID- 17917418 TI - [Stereochemistry, syntheses and biological activity of lupine alkaloids--from studies on the leguminous plants growing mainly in Japan]. AB - Lupine alkaloids have been studied from the viewpoints of biosynthesis, biotechnology, chemotaxonomy, and biological activity, on the basis of the chemical investigation of the leguminous plants of the 28 species belonging to the 9 genera, which mainly grow in Japan. The results obtained have been comprehensively reviewed by authors. This review describes the stereochemistry of lupine alkaloids and focuses on the conformational flexibility of nitrogen-fused systems such as quinolizidine and indolizidine, syntheses of new unusual types of alkaloids from known lupine alkaloids, and pharmacological activity of lupine alkaloids, especially kappa-opioid receptor-mediated antinociceptive effects of matrine-type lupine alkaloids. PMID- 17917419 TI - [Homocysteine metabolism]. AB - Homocysteine, a sulfur amino acid, is an intermediate metabolite of methionine. In 1969, McCully reported autopsy evidence of extensive arterial thrombosis and atherosclerosis in children with elevated plasma homocysteine concentrations and homocystinuria. On the basis of this observation, he proposed that elevated plasma homocysteine (hyperhomocysteinemia) can cause atherosclerotic vascular disease. Hyperhomocysteinemia is now well established as an independent risk factor for atherosclerotic vascular disease. Mild hyperhomocysteinemia is quite prevalent in the general population. It can be caused by genetic defects in the enzymes involved in homocysteine metabolism or nutritional deficiencies in vitamin cofactors, certain medications or renal disease. An increase of 5 micromol per liter in the plasma homocysteine concentration raises the risk of coronary artery disease by as much as an increase of 20 mg per deciliter in the cholesterol concentration. In this article, we review the biochemical, experimental and clinical studies on hyperhomocysteinemia, with emphasis on the metabolism and pharmacokinetics of homocysteine. PMID- 17917420 TI - [Pharmacognosical study during 40 years]. AB - In order to quality control for medicinal plants such as Aconitum charmicaelii, Rhemannia glutinosa, Atractyrodes lancea, Pinellia ternata, Panax species and Gentiana scabra were clonally micropropagated by tip tissue culture and embryogenetic techniques. Monoclonal antibodies against the bioactive compounds contained in the crude drugs were prepared and set up the ELISA as a high sensitive and quick determination method. A newly developed eastern blotting methodology can stain typically the only antigen molecule and the related compounds having a same aglycone. Knockout extract can be prepared by using an immunoaffinity column conjugated with monoclonal antibody, and its importance has been discussed. Single chain fragment-variable gene against solamargine was cloned and transformed to a host plant, Solanum chasianum resulting in the increase of antigen molecule as 2.5 to 3 times. Three biosynthetic enzymes regarding marihuana compounds, THCA-, CBDA-, CBCA-synthases were isolated. THCAsynthase was cloned, over expressed and confirmed its characteristics including FAD combining enzyme, and finally determined the structure by X-ray analysis. The distribution of THCAsynthase was also investigated using its and GFP hybrid gene. We found new functions for saffron like improving learning and memory and LTP for blocking by ethanol. A folk medicine in Taiwan, Anoectochilus formosanus was propagated in vitro and investigated opening new pharmacological activities in lipid metabolism. PMID- 17917421 TI - [Derivative spectrophotometric and NMR spectroscopic study in pharmaceutical science]. AB - This review starts with an introduction of derivative spectrophotometry followed by a description on the construction of a personal computer-assisted derivative spectrophotometric (DS) system. An acquisition system for inputting digitalized absorption spectra into personal computers and a BASIC program for calculating derivative spectra were developed. Then, applications of the system to drug analyses that are difficult with traditional absorption methods are described. Following this, studies on the interactions of drugs with biological macromolecules by the DS and NMR methods were discussed. An (1)H NMR study elucidated that the small unilamellar vesicle (SUV) has a single membrane made of a phosphatidylcholine bilayer, and that chlorpromazine interacts with both the outer and inner layers. (13)C NMR revealed a reduction of the dissociation constants of phenothiazine drugs due to their interaction with SUV. The partition coefficients of phenothiazine, benzodiazepine and steroid drugs in an SUV-water system and the effects of cholesterol or amino lipids content on these partition coefficients were examined by the DS method. The binding constants of phenothiazine drugs to bovine serum albumin (BSA) and the influence of Na(+), K(+), Cl(-), Br(-), and I(-) on these binding constants were determined by DS. It was found that I(-), Br(-), Cl(-) reduce the binding constants in this order, and that Na(+) and K(+) have no effect. A (19)F NMR study revealed that triflupromazine binds to BSA and human serum albumin in two regions including Site II with different populations, and that a nonsteroidal anti-inflammatory drug, niflumic acid, binds Sites Ia and Ib. PMID- 17917422 TI - [Molecular pharmacologic approaches to functional analysis of new biological target molecules for drug discovery]. AB - This review focuses on two pharmacologic approaches to the functional evaluation of new target molecules for drug discovery. One is the development of a novel specific antagonist of the Na(+)-Ca(++) exchanger (NCX) SEA0400. The other is a comprehensive analysis of the functions of pituitary adenylate cyclase-activating polypeptide (PACAP), a neuropeptide ligand for G protein-coupled receptors. NCX is the one of the last target molecules regulating the cellular Ca(++) concentration. There was no efficient way to address the pathophysiologic roles of NCX until a specific antagonist, 2-[4-[(2,5-difluorophenyl)methoxy]phenoxy]-5 ethoxyaniline (SEA0400), was developed. Our recent studies using SEA0400 clearly showed the possible roles of NCX in several pathologic states of cardiovascular and nervous tissues. In our second approach including gene-targeting methods, we found new, unexpected roles of PACAP in higher brain functions, such as psychomotor, cognition, photoentrainment, and nociception. Based on these experimental findings, a genetic association study in schizophrenia patients revealed that the single-nucleotide polymorphisms of the PACAP gene are significantly associated with the hypofunction of the hippocampus. Regarding the peripheral roles of PACAP, we found that PACAP is involved not only in the regulation of insulin secretion in pancreatic islets, but also in the regulation of islet turnover. In subsequent phenotypic analysis of PACAP transgenic mice, we identified novel candidate genes that probably have promising functional roles. PMID- 17917423 TI - [Development of multifunctional envelope type artificial viral-like gene delivery system]. AB - This review introduces a new concept "Programmed Packaging" to develop a non viral gene delivery system. Based on this concept, multifunctional envelope type nano devices (MEND) were developed for in vitro, in situ and in vivo conditions. A quantitative study to identify a rate limiting step in intracellular trafficking was also shown between viral and non-viral vectors, which indicated an important role of controlled intranuclear disposition for development a safe and efficient non-viral gene delivery system. This review will provide a future direction of non-viral gene delivery system. PMID- 17917424 TI - [Structural and functional studies on proteins as potential drug discovery targets]. AB - Structural biology teaches us about the fundamental nature of biological molecules. Drug design is the most immediate medical application of structural biology. Therefore our studies have been focused on structural and functional studies of human disease-related proteins and proteins essential for the growth and development of pathogenic organisms. The present paper describes 1) structural biological studies of human autocrine motility factor, 2) structural biological studies of human ribonuclease L, and 3) structural biological studies of Plasmodium falciparum S-adenosyl- L-homocysteine hydrolase. PMID- 17917425 TI - [Development of a novel artificial gene delivery system multifunctional envelope type nano device for gene therapy]. AB - For efficient gene delivery to the nucleus, nonviral vectors need to overcome several barriers such as the plasma membrane, endosomal membrane, and nuclear membrane. To overcome these obstacles, it is necessary to equip the delivery system with various functional devices. However, it is difficult to package all such functional devices into a single system to exert each of their functions at the appropriate time and at the correct location. Thus our group proposed a new packaging concept, "programmed packaging." A multifunctional envelope-type nano device (MEND) was developed for use as an efficient nonviral system for the delivery of plasmid DNA (pDNA), oligodeoxynucleotide (ODN), and siRNA using octaarginine (R8) as an internalizing ligand based on the programmed packaging. The R8-modified MEND (R8-MEND) encapsulating pDNA showed significantly high transfection activity comparable to that of adenovirus, and the uptake pathway of R8-MEND was macropinocytosis, which can avoid lysosomal degradation. R8-MEND successfully delivered gene to hair follicles after in vivo topical application to mouse skin. Moreover, R8-MEND encapsulating anti-luciferase ODN using protamine showed a 90% antisense effect, and R8-MEND encapsulating siRNA condensed with stearylated R8 significantly silenced luciferase activity. Our group thus succeeded in the development of R8-MEND based on programmed packaging, and MEND is a promising new delivery system for pDNA and functional nucleic acids. PMID- 17917426 TI - [Endeavors to make the photophore, diazirine easy to use]. AB - Photoaffinity labeling enables the direct probing of a target protein through a covalent bond between a ligand and its binding protein. We used carbene generating phenyldiazirine as a photophore because practical examinations had shown that the phenyldiazirine functioned as the powerful barb on the hook. This review describes improvements of synthetic strategies of the photoaffinity ligands bearing diazirine. First, we dramatically improved the direct formylation of phenyldiazirine, which was a practical diazirine source, to obtain a versatile diazirine unit. Second, we established "the photoreactive unit technique" for a one-step introduction of phenyldiazirine into peptides, proteins, DNAs, and sugars. Since the photoreactive units can be easily integrated into physiological ligands by chemoselective reaction, the biochemists, who are not familiar with organic synthesis, can prepare the photoaffinity ligands using their interested ligands. Our improvements would promote the utilization of phenyldiazirine for analyzing biological interfaces, and extend the potential of photoaffinity labeling as a sensitive means of rapidly elucidating protein structures and proteomic profiling. PMID- 17917427 TI - Effect of Anwala churna (Emblica officinalis GAERTN.): an ayurvedic preparation on memory deficit rats. AB - The present study was aimed at investigating the effects of Anwala churna (Emblica officinalis GAERTN.), an Ayurvedic preparation, on memory in rats. Anwala churna was administered orally in three doses (50, 100 and 200 mg/kg) for 15 days to different groups of young and aged rats. The elevated plus-maze and Hebb-Williams maze served as exteroceptive behavioral models for testing memory. Diazepam-, scopolamine-, and ageing induced amnesia served as the interoceptive behavioral models. Anwala churna (50, 100, and 200 mg/kg, p.o.) produced a dose dependent improvement in memory scores of young and aged rats. Furthermore, it reversed the amnesia induced by scopolamine (0.4 mg/kg, i.p.) and diazepam (1 mg/kg, i.p.). Based on these results, Anwala churna may prove to be a useful remedy for the management of Alzheimer's disease due to its multifarious beneficial effects such as memory improvement and reversal of memory deficits. PMID- 17917428 TI - Pharmacokinetics of piperaquine after single and multiple oral administrations in healthy volunteers. AB - The aim of this work was to study the pharmacokinetics of piperaquine in healthy volunteers. Healthy volunteers received piperaquine and tablets of Artekin by oral administration. The plasma samples were analyzed for piperaquine by liquid liquid extraction and determined by HPLC-UV. The results demonstrated that the plasma drug concentration-time curves of single and multiple dose of piperaquine were fitted to a two-compartment open model. The pharmacokinetics parameters of piperaquine alone in a single dose were: t(1/2(beta))=(317.2-/+126.6)h, AUC(0- >infinity)=(44293-/+12636)h x ng/ml, V(d)=(9490.9-/+2161.9)ml/kg, and Cl=(22.83 /+9.83)ml/h/kg. In Artekin in a single dose these parameters were: t(1/2(beta))=(302.8-/+180.7)h, AUC(0-->infinity)=(46419-/+13670)h x ng/ml, V(d)=(10188.6-/+3520.3)ml/kg, and Cl=(25.48-/+10.89)ml/h/kg, while in Artekin in multiple doses they were: t(1/2(beta))=(298.9-/+101.9)h, AUC(0- >infinity)=(227692-/+56294)h x ng/ml, V(d)=(5031.5-/+1097.8)ml/kg, Cl=(11.91 /+3.046)ml/h/kg, respectively. The absorption and distribution of piperaquine were quick while the elimination was quite slow. There were significant differences in the pharmacokinetics parameters of piperaquine in Artekin between a single dose and multiple doses (p<0.001), suggesting that piperaquine might accumulate in vivo and that attention should be given to its possible adverse drug reactions in clinical treatment. PMID- 17917429 TI - Hypoglycemic effects and mechanisms of action of Cortex Lycii Radicis on alloxan induced diabetic mice. AB - Cortex Lycii Radicis (CLR) has been used as a traditional Oriental medicine as an antipyretic and to treat pneumonia, night-sweats, cough, hematemesis, inflammation, and diabetes mellitus for centuries. This study aimed to determine the effects of CLR on alloxan-induced diabetic mice and its mechanisms. Based on thin-layer chromatography (TLC) assay, the main compounds of CLR include an organic acid, flavone, alkaloid, polysaccharide, anthraquinone, and saponin. The mice were divided into four groups: normal control (NC), diabetes control (DC), diabetes+high-dose CLR (200 mg kg(-1)), and diabetes+low-dose CLR (100 mg kg( 1)). The diabetic mice were administered CLR daily for 28 days. The CLR treatment resulted in significant decreases in fasting blood glucose, total cholesterol, and triglycerides. CLR also showed a tendency to improve body weight gain in diabetic mice. Furthermore, the serum insulin level of each group was assayed, and the DC group had a lower serum insulin level than the NC group. Insulin levels were dose dependently raised in the CLR-treated groups compared with the DC group. According to single-cell gel electrophoresis and LD(50) analysis, CLR was nontoxic to the animals. The results indicate that CLR alleviates the blood glucose and lipid increases associated with diabetes and improves the abnormal glucose metabolism and increases insulin secretion by restoring impaired pancrease beta-cells in alloxan-induced diabetic mice. The results suggest that CLR has hypoglycemic potential and could be useful in diabetes therapy. PMID- 17917430 TI - Determination of paracetamol and orphenadrine citrate in pharmaceutical tablets by modeling of spectrophotometric data using partial least-squares and artificial neural networks. AB - The estimation of paracetamol and orphenadrine citrate in a multicomponent pharmaceutical dosage form by spectrophotometric method has been reported. Because of highly interference in the spectra and the presence of non-linearity caused by the analyte concentrations which deviate from Beer and Lambert's law, partial least-squares (PLS) and artificial neural networks (ANN) techniques were used for the calibration. A validation set of spiked samples was employed for testing the accuracy and precision of the methods. Reasonably good recoveries were obtained with PLS for paracetamol and the use of an ANN allowed the estimation of orphenadrine citrate, a minor component which could not be adequately modeled by PLS. Three production batches of a commercial sample were analysed, and there was statistically no significant difference (P<0.05) between the results with the proposed method and those obtain with the official comparative method. PMID- 17917431 TI - [Assessment of compliance for oral medicines with MMSE, Mini-Mental State Examination, in hospitalized elderly patients]. AB - The minimental state examination (MMSE) is a widely used, standardized method to assess cognitive function including movement-related disorders with high reliability. We studied the relationship between MMSE scores and the ability to take oral medications correctly (ingestion compliance) in 70 elderly inpatients (mean age 71.3+/-7.0 years). Patients with abnormal glucose tolerance as determined by an HbA(1c) level of 5.8% or greater including diabetes showed a trend of lower MMSE scores compared with patients with normal glucose tolerance, and the scores were negatively correlated with HbA1c, age, and systolic blood pressure (P<0.05). Self-management in taking oral medications was very difficult in 4 patients whose MMSE scores were 21 points or less. Thus ingestion supervisions by nurses were required in these patients. Furthermore, 9 of 12 noncompliant patients had MMSE scores ranging from 22 to 26 points. We instructed these patients to take medications in a one-dose package as a useful tool to improve compliance. The MMSE score was 27 or higher in 44 of 54 compliant patients, and 10 patients had scores ranging from 21 to 26. The sensitivity and specificity for noncompliance at an MMSE score cut-off point of 26 were 75.0% and 81.5%, respectively. In conclusion, it is necessary to coordinate ingestion methods matched to each patient according to their abilities to comply with medication schedules. They should be preevaluated with the MMSE to improve ingestion compliance. The MMSE is a recommended test in hospitalized elderly patients for the assessment of the ability to take medications safely. PMID- 17917432 TI - In vitro corneal permeation of diclofenac from oil drops. AB - In vitro transcorneal permeation of diclofenac from oil drops was studied using freshly excised goat cornea. The maximum apparent corneal permeability coefficient (Papp) was obtained with 0.2% (w/v) diclofenac drops in sesame oil followed by safflower oil, while formulation in castor oil provided minimal Papp. The addition of benzyl alcohol, a preservative, in oil drops, increased the Papp value of diclofenac. Partition experiments indicated increased partitioning of diclofenac in the aqueous phase in the presence of benzyl alcohol, and the same could be responsible for the benzyl alcohol-induced increase in Papp. The solubility of diclofenac was higher in castor, arachis, and sunflower oil. But drug permeation from 0.5-1.0% (w/v) diclofenac drops in castor oil or 0.5% (w/v) drops in arachis /sunflower oil was less than that observed with 0.2% (w/v) drops in sesame oil. Thus diclofenac 0.2% (w/v) drops in sesame oil containing 0.5% (v/v) benzyl alcohol provides maximum Papp. The formulation increased corneal hydration indicating corneal damage. Since corneal hydration is less than 83% the damage appears to be reversible. The saturation solubility of diclofenac in sesame oil at 4 degrees C is 0.33% (w/v). Hence diclofenac 0.2% (w/v) solution in sesame oil will not precipitate at 4 degrees C and therefore the chances of crystallization of diclofenac from the formulation due to climatic change leading to physical instability appear to be remote. PMID- 17917433 TI - Lipids behavior and adverse effects for oral antidiabetic agents in patients with Type 2 diabetes treated with sulfonylureas alone based on systematic review. AB - The secondary and adverse effects when biguanides, alpha-glycosidase inhibitor or thiazolidine derivative was used with sulphonylurea agent (SU) as compared with those with SU alone in Type 2 diabetes patients by using Systematic Review. Two agent concurrent treatment groups, taken from studies in which subjects were assigned to a group given only a sulfonylurea agent and a group given a sulfonylurea agent with the other glycemic control agent (combination of a sulfonylurea agent and a biguanide agent (I), combination of a sulfonylurea agent and an alpha-glucosidase inhibitor (II), and combination of a sulfonylurea agent and thiazolidinedione (III)), were studied in a randomized controlled trial. The secondary efficacy outcome measures were total cholesterol (TC), triglyceride (TG), HDL-C, LDL-C, and change in body weight. The incidence of hypoglycemia, feeling of fullness, diarrhea, liver dysfunction, and edema was investigated as a safety outcome measure, and the clinical significance of concurrent treatment with a sulfonylurea agent in addition to the other glycemic control agent was investigated. With respect to (II), an antidiabetic effect was showed. As for (III), it had the disadvantage of increased body weight. Furthermore, increase of HDL-C levels, in particular, was observed. The improving effect of (III) on serum lipids may be clinically effective for considering the pathologic condition of diabetes, which is often complicated by hyperlipidemia. PMID- 17917434 TI - Synthesis of some novel oxadiazole and oxadiazoline analogues for their antiinflammatory activity. AB - The search for newer non-steroidal antiinflammatory drugs (NSAIDs) and the importance of oxadiazoles as antiinflammatory agents prompted us to undertake the synthesis of some novel oxadiazole and related analogues with unreported antiinflammatory activities. The antiinflammatory potential of the compounds was investigated using the carrageenan-induced rat paw edema method and cotton pellet induced granuloma method. Some compounds demonstrated marked antiinflammatory activities. The antiinflammatory activity of oxadiazoles at doses of 100 mg/kg was shown by their ability to provide 28-55%, 21-36%, and 27-49% protection against carrageenan-induced rat paw edema, moist cotton pellet-induced, and dry cotton pellet-induced granuloma, respectively. On the other hand, the antiinflammatory properties of oxadiazolines at doses of 100 mg/kg was reflected by their ability to provide 15-47%, 22-39%, and 23-47% protection against carrageenan-induced rat paw edema, moist cotton pellet-induced, and dry cotton pellet-induced granuloma, respectively. Structure-activity relationships among synthesized compounds were also studied. PMID- 17917435 TI - [Usefulness of the classification technique of cerebral artery for 2D/3D registration.]. AB - Several papers have proposed 2D/3D registration methods of the cerebral artery using magnetic resonance angiography (MRA) and digital subtraction angiography (DSA). Since differences between vessels in a DSA image and MRA volume data cause registration failure, we previously proposed a method to extract vessels from MRA volume data using a technique based on classification of the cerebral artery. In this paper, we evaluated the usefulness of this classification technique by evaluating the reliability of this 2D/3D registration method. This classification method divides the cerebral artery in MRA volume data into 12 segments. According to the results of the classification, structures corresponding to vessels on a DSA image can then be extracted. We applied the 2D/ 3D registration with /without classification to 16 pairs of MRA volume data and DSA images obtained from six patients. The registration results were scored into four levels (Excellent, Good, Fair and Poor). The rates of successful registration (> fair) were 37.5% for registration without classification and 81.3% for that with classification. These findings suggested that there was a low percentage of incorrectly extracted voxels and we could facilitate reliable registration. Thus, the classification technique was shown to be useful for feature-based 2D/ 3D registration. PMID- 17917436 TI - Analysis of the effective point of measurement of a thimble chamber dosimeter set parallel to the X-ray beam axis. AB - To measure the narrow beam used in stereotactic irradiation, installation of the ionization chamber parallel to the X-ray beam axis has been used instead of perpendicular installation. However, the definition of the effective point is a major problem in the parallel installation. In this study, we analyzed the effective point in parallel installation, and considered the prediction and evaluation of measurement point displacement. Relative dosimetry was carried out by installing the thimble ionization chamber in both perpendicular and parallel configurations. We then searched for the measurement point that coincided with the PDD of the perpendicular installation by using the displacement of the measurement point of the parallel installation. We found that the effective point of measurement for relative photon beam dosimetry depends on every detail of the chamber design, including the cavity length and the cavity radius. Moreover, the effective point of measurement also depends on the beam quality and the field size. The amount of effective point displacement for the parallel installation was quantified with the linear expression of TPR(20,10). Our results showed that the amount of effective point displacement can be estimated by the ionization volume of the dosimeter and the energy used. PMID- 17917437 TI - [Enactment of TR14969:2007-Quality Management System for Medical Devices; Guide to the Application of JIS Q 13485:2005.]. PMID- 17917438 TI - Kuwait acute coronary syndromes registry: baseline characteristics, management practices and in-hospital outcomes of patients hospitalized with acute coronary syndromes in Kuwait. AB - OBJECTIVES: To identify the characteristics of patients with acute coronary syndromes (ACS), their hospital management and in-hospital outcomes, through a prospective registry system in Kuwait. SUBJECTS AND METHODS: A registry involving all 7 general hospitals in Kuwait was set up. Consecutive patients diagnosed as having ACS over a period of 6 months were enrolled. RESULTS: Of 2,129 patients enrolled, 718 (34%) had ST segment elevation myocardial infarction (STEMI), 576 (27%) non-ST segment elevation myocardial infarction (NSTEMI) and 835 (39%) unstable angina (UA). Thrombolytic therapy was used in 556 (77%) patients with STEMI. The median time from diagnostic electrocardiogram to administration of thrombolytic therapy was 38 min. Almost all patients with ACS (2,050, 96%) received aspirin during hospitalization. Only a minority received clopidogrel, 18 (3%) STEMI, 36 (6%) NSTEMI and 96 (12%) UA patients. The use of glycoprotein IIb/IIIa antagonists was minimal (38 patients, 2%). beta-Blockers were used in 1,473 (69%) patients, while 982 (46%) received angiotensin-converting enzyme inhibitors. Coronary angiography during hospitalization was performed in 119 (17%), 120 (21%) and 126 (15%) patients with STEMI, NSTEMI and UA, respectively. In-hospital mortality occurred in 31 (4%) myocardial infarction patients and 4 (0.5%) UA patients (p < 0.0001). CONCLUSION: This registry has enabled us to determine the incidence and characteristics of ACS patients in Kuwait. It has also enabled us to identify some barriers that we need to overcome for the full implementation of published guidelines for the management of patients with ACS. PMID- 17917439 TI - Barriers to seeking preventive dental care by Kuwaiti adults. AB - OBJECTIVE: The aim of this study was to assess the prevalence of preventive dental visits and to identify self-reported barriers for this practice among Kuwaiti adults. MATERIALS AND METHODS: A self-administered, anonymous, structured questionnaire was distributed to a random sample of Kuwaiti nationals 18 years of age or older recruited from all six health districts of Kuwait. A total of 2,400 questionnaires were distributed. Multiple logistic regression analysis was performed to identify factors independently associated with not having a preventive dental visit for more than 1 year. RESULTS: Of the 2,400 questionnaires, 1,925 (80.2%) were completed. Of these, 620 (32.2%) had a dental visit within the previous 6 months, 504 (26.2%) between 6 and 12 months and 801 (41.6%) more than 12 months ago. The most common reasons for the last dental visits were pain or a dental emergency, need for restorative treatment, and an examination/prophylaxis. The strongest factors for not having preventive visits were not using a mouthrinse daily, flossing less than once a day, dental fear, belief that there is no need for visits unless pain was present, brushing the teeth less than twice a day, and believing that appointments are too far ahead. Also older respondents (>30 years), female gender, and those having only high school education or less were less likely to visit a dentist for preventive reasons. CONCLUSION: More than half of the studied population reported not having had a preventive visit for more than 1 year. Unfavorable self-care habits, dental fear and belief that visiting a dentist is necessary only for pain relief were the strongest factors for the nonattendance behavior. PMID- 17917440 TI - Essential trace elements and antioxidant status in relation to severity of HIV in Nigerian patients. AB - OBJECTIVES: This study was designed to determine the plasma levels of some antioxidants and trace elements in three severity groups of HIV patients compared with non-HIV-infected controls. METHODS: The plasma levels of antioxidants (total antioxidant, albumin, bilirubin and uric acid) and trace elements (Mg, Fe, Zn, Mn, Cu, Cr, Cd and Se) were estimated spectrophotometrically in controls and patients with CD4 counts of <200; 200-499 and > or =500 cells/microl. RESULTS: Uric acid and Zn were significantly higher, while vitamin E and all the trace elements (except Zn) were significantly lower in HIV-infected patients compared to healthy controls. The highest level of uric acid was observed in those with CD4 counts of <200 cells/microl. All the trace elements (except Zn) were higher in HIV subjects with a CD4 count of 200-499 cells/microl compared to >500 cells/microl. Only uric acid and Zn showed significant correlation with CD4 count. CONCLUSION: Based on the results of this study, we recommend routine assessment and appropriate supplementation of antioxidants/trace elements in HIV subjects. This supplementation is hoped to strengthen the immune system and reduce the adverse consequences of HIV- related oxidative stress. PMID- 17917441 TI - Comparative bioavailability of norfloxacin tablets based on blood and urine data. AB - OBJECTIVE: To assess the bioavailability of norfloxacin from urinary excretion relative to plasma concentration. MATERIALS AND METHODS: Twelve healthy volunteers (22-33 years) participated in the study. Each received a previously developed (M), a local (L) and a multinational (Noroxin) tablet (Ref), 400 mg each, according to a random balanced three-way crossover design on 3 different days. Blood samples were collected over a 12-hour period and urine over a 24-hour period. Norfloxacin concentrations were analyzed by a validated HPLC method. RESULTS: An initial estimate of bioequivalence of the three products was obtained using analysis of variance on transformed data and based on confidence interval calculation. Elimination pharmacokinetic parameters (half-life and renal clearance) calculated from plasma concentration and urinary excretion data (mean values, n = 36) were comparable to reported values for norfloxacin. Interproduct differences in elimination parameters (mean values, n = 12) were statistically insignificant (F values, ANOVA). Strong association was found between the mean of plasma concentration and urinary excretion rates for many volunteers (F values, regression analysis). Relative bioavailability values calculated for the local and previously developed products relative to Noroxin were higher than 85% based on area under the curve and urinary excretion. Bioequivalence could not be established among the three tested products based on calculated 90% confidence intervals. CONCLUSION: Urinary excretion of norfloxacin may be a useful noninvasive tool for bioavailability assessment of norfloxacin oral formulations. PMID- 17917442 TI - Plasma thiobarbituric acid-reactive substance levels in subclinical hypothyroidism. AB - OBJECTIVE: The purpose of this study was to determine thiobarbituric acid reactive substance (TBARS) levels in subclinical hypothyroidism and to examine the effect of levothyroxine replacement on TBARS levels. SUBJECTS AND METHODS: A cohort of 28 female patients with subclinical hypothyroidism and 24 healthy controls were enrolled in this study. The levels of plasma TBARS, serum lipids, and high-sensitive C-reactive protein (CRP) in patients with subclinical hypothyroidism at baseline and after achieving euthyroid state by levothyroxine were assessed. RESULTS: TBARS levels of the patients were similar to those of the control group in the subclinical hypothyroid state and after restoration of euthyroidism by levothyroxine replacement. TBARS levels decreased after levothyroxine treatment, but did not reach statistical significance. There was no significant correlation between TBARS, lipid and CRP levels. Serum CRP levels were higher in subclinical hypothyroidism (4.28 +/- 0.9 mg/l) than in the control group (1.95 +/- 0.34 mg/l) and the difference was statistically significant (p = 0.03). After achieving euthyroid state, CRP levels decreased significantly in patients with subclinical hypothyroidism from 4.28 +/- 0.9 to 2.32 +/- 0.6 mg/l (p = 0.006). CONCLUSION: Our findings suggest that there is no significant alteration of plasma TBARS levels neither in subclinical hypothyroid state nor after achieving euthyroid state. Serum CRP level is higher in patients with subclinical hypothyroidism than in the control group. Normalization of thyroid state seems to effectively reduce serum CRP levels in subclinical hypothyroidism without any correlation with TBARS activity. PMID- 17917443 TI - Effects of programs for prevention of early childhood caries. A systematic review. AB - OBJECTIVE: To determine the most effective intervention program for prevention of early childhood caries. MATERIALS AND METHODS: All studies published after 1966 were identified by searching electronic databases (Medline, The Cochrane Library, Embase, Dissertation and Serfile databases) and manual searching. Studies were included if they analyzed the effect of an intervention to prevent caries in 0- to 5-year-old children, recorded caries as dmfs, dmft, dfs or dft, and utilized a randomized controlled study design. RESULTS: The initial search produced 916 citations, 19 of which met the inclusion criteria. The quality of the articles was assessed independently by the reviewers based on randomized selection and double blinding. The 7 articles included in the actual review evaluated the effects of dental health education, prenatal fluoride administration, topical fluoride application, preventive dental programs, and use of fluoridated toothpaste. Meta-analysis was not possible because of differences in the types of intervention among the studies. CONCLUSION: Conclusive evidence regarding the best intervention to prevent early childhood caries could not be drawn due to the flawed design of somepapers. However, fluoride-based interventions appear to be effective in young children. More studies are required to determine the best method of delivery. PMID- 17917444 TI - Antinociceptive, anti-inflammatory and antipyretic properties of the aqueous extract of Bauhinia purpurea leaves in experimental animals. AB - OBJECTIVE: The present study was carried out to determine the antinociceptive, anti-inflammatory and antipyretic activities of the aqueous extract of Bauhinia purpurea leaves using animal models. MATERIALS AND METHODS: The air-dried, powdered leaves (approximately 20 g) were soaked in distilled water (1:20; w/v) at room temperature for 72 h and the supernatant obtained was freeze-dried. The crude dried extract (approximately 2.4 g) was prepared in doses of 6.0, 30.0 and 60.0 mg/kg, and subjected to the respective antinociceptive (abdominal constriction, hot plate and formalin tests), anti-inflammatory (carrageenan induced paw edema test) and antipyretic (brewer's yeast-induced pyrexia test) assays. RESULTS: The results obtained indicate that the extract possessed significant (p < 0.05) antinociceptive, anti-inflammatory and antipyretic activities, which were not dependent on the doses of extract used. The highest concentration of extract was less effective as an anti-inflammatory and an antipyretic agent. CONCLUSION: This study showed that the aqueous extract of B. purpurea leaves possesses potential pharmacological activities that require further investigation and, thus, confirms the folklore use of the plant in the treatment of ailments associated with pain and inflammation. PMID- 17917445 TI - Adenotonsillar granuloma: histopathological correlation. AB - OBJECTIVES: This study was undertaken to investigate the presence and nature of granulomatous changes in the adenotonsillar tissues and their relation to systemic disorders. MATERIALS AND METHODS: The study was a retrospective clinicopathological review; 21,410 specimens of tonsil and adenoid tissue were subjected to histological examination during the period from 1995 to 2003. Twenty three cases of tonsillar and adenoid granuloma (12 males and 11 females) were identified. Haemotoxylin and eosin stains were reviewed and clinical features noted. RESULTS: Of 23 cases, 11 were adenotonsillectomies, 10 tonsillectomies, 1 adenoidectomy and 1 excision biopsy of the right tonsil. Histologically, the slides reviewed showed 4 types of granulomatous inflammation that can affect the adenotonsillar tissues. The most common type was non-caseating epithelioid granulomas. Clinically, 22 cases presented with symptoms of chronic tonsillitis, nasal obstruction and obstructive sleep apnoea. None had any systemic granulomatous disorders. Only 1 case had tuberculosis. CONCLUSION: In all the cases but one there was no obvious systemic aetiology for the granulomatous changes in the adenotonsillar tissues. Hence, the cost effectiveness of clinicopathological examination of the resected adenoid and tonsil is still questionable. PMID- 17917446 TI - Investigation of proliferative activity in the developing human tooth using Ki-67 immunostaining. AB - OBJECTIVE: The aim of this study was to investigate the proliferation of the developing human tooth germ and its surrounding tissues using Ki-67 immunostaining. MATERIALS AND METHODS: Sections of mandibular dental arch tissues collected from 4 cadaveric human fetuses of 13, 16, 21 and 30 weeks of gestation were used. The immunoreactivity of Ki-67 in the tissue sections was assessed visually under a light microscope. Immunohistochemical controls were performed by replacing the primary antibody with phosphate-buffered saline or normal rabbit lgG. RESULTS: The control sections did not display Ki-67 immunoactivity. Specimens of 13 weeks of gestation revealed intense Ki-67 immunostaining throughout the entire developing mandibular primary molars. At 16 weeks of gestation, immunostaining was observed in the inner enamel epithelium and dental papilla, in conjunction with the dental lamina showing decreased immunostaining. At 21 weeks, Ki-67 immunostaining was observed only in the inner enamel epithelium and dental papilla. The immunoreactivity of active ameloblasts and odontoblasts decreased, along with the proliferation capacity of the dental lamina. At 30 weeks, both enamel and dentin formation was observed along the cusped aspect of the tooth germ. Ameloblasts and odontoblasts were no longer immunoreactive in this region, while both types of cells were immunoreactive at the cervical regions of the crown. Dental lamina cells showed disintegration and were totally Ki-67-negative at 30 weeks of gestation. CONCLUSION: The Ki-67 immunoreactivity of the dental lamina decreased during intrauterine tooth development. Positive immunostaining was observed at specific sites in the enamel organ and dental papilla during the cap and bell stages. PMID- 17917447 TI - Cutaneous metastases from squamous cell carcinoma of the ureter. AB - OBJECTIVE: To present an extremely rare case of squamous cell carcinoma of the ureter with cutaneous metastases. CLINICAL PRESENTATION AND INTERVENTION: A case is presented involving a 67-year-old woman presenting with a clinical history of squamous cell carcinoma of the ureter and who had undergone a nephro-ureterectomy with a bladder cuff excision in May 2004. The pathologic report showed squamous differentiation, as well as keratin pearl formation. A large regional cutaneous lesion on the chest wall was found in January 2006, and a biopsy showed metastatic malignant urothelial tumors consisting of squamous cell carcinomas. CONCLUSION: This report describes a case of cutaneous metastasis from a squamous cell carcinoma of the ureter that is extremely rare with a generally dismal prognosis. PMID- 17917448 TI - Ochrobactrum anthropi bacteremia in a child with inborn error of mitochondrial fatty acid oxidation. AB - OBJECTIVE: To report an incident of bacteremia caused by Ochrobactrum anthropi. CASE PRESENTATION AND INTERVENTION: The case of a female child aged 2 years and 10 months with a known history of long-chain 3-hydroxyacyl-coenzyme A dehydrogenase deficiency who developed O. anthropi bacteremia during hospital stay is presented. Patient's history, clinical findings, laboratory and radiological investigations were thoroughly reviewed. The cultured organism was identified using MicroScan WalkAway 96 SI (Dade Behring) as well as by conventional techniques. Imipenem resistance was confirmed by the conventional Kirby-Bauer disk diffusion technique on Muller-Hinton agar with no zone of inhibition around a 10-mug imipenem disk (Hi Media) using the 0.5 McFarland standard. CONCLUSION: This report shows O. anthropi as a rare nosocomial pathogen that affected a patient who was immunocompromised. The O. anthropi showed multidrug resistance. PMID- 17917449 TI - The science and art of medicine. PMID- 17917450 TI - The reform of medical education. PMID- 17917451 TI - The past and future of neuroprotection in cerebral ischaemic stroke. AB - Following the realization that cerebral tissue may survive for hours after an ischaemic insult, several agents with neuroprotective properties in small-animal models of cerebral ischaemia have been tested in patients with acute ischaemic stroke (AIS). Initial attempts at translating the positive effects of these agents from animal models to patients were unsuccessful, possibly as a result of poorly planned experiments in models of ischaemia, and/or clinical trials of AIS that were not optimized to show a positive effect. Newer neuroprotective agents that are believed to act later in the ischaemic cascade may offer a greater chance of success. However, before these agents can be introduced into clinical practice they must undergo assessment in rodent and large-animal models of AIS and, in particular, the dose-response relationship and the treatment time window should be defined. This should be followed by evaluation in carefully designed clinical trials of adequate power, involving subjects receiving an appropriate dose within an optimum time window following the onset of symptoms. There is hope that such careful strategies may guide continued progress in neuroprotective drug development. PMID- 17917452 TI - Association study of human MTH1 Ile45Thr polymorphism with sporadic Parkinson's disease. AB - Human MTH1, an oxidized purine nucleoside triphosphatase, hydrolyzes 8-oxo-dGTP thereby preventing its misincorporation into DNA. The present study was designed to investigate a possible link between the MTH1 Ile45Thr polymorphism and the development of sporadic Parkinson disease (PD). This case-control study consisted of 106 PD patients and 135 unrelated controls. MTH1 polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The results showed that Ile45/Thr45 heterozygote and Thr45 allele tended to be more frequent in sporadic PD, although statistically not significant (0.085 vs. 0.044, corrected p = 0.591 and 0.052 vs. 0.022, p = 0.080, respectively). Stratification analysis by gender showed that Ile45/Thr45 heterozygote tended to be more frequent in male PD patients than in male controls (0.113 vs. 0.038, corrected p = 0.480). The male PD patients exhibited a borderline statistically significant higher frequency of the Thr45 allele than the controls (0.073 vs. 0.019, corrected p = 0.050). These results suggested to us that the Thr45 allele of MTH1 might be associated with sporadic PD in the Chinese male population. PMID- 17917453 TI - Purkinje cell loss in the cerebellar flocculus in patients with ataxia with ocular motor apraxia type 1/early-onset ataxia with ocular motor apraxia and hypoalbuminemia. AB - We genetically screened patients with ataxia with ocular motor apraxia type 1 (AOA1)/early-onset ataxia with ocular motor apraxia and hypoalbuminemia (EAOH), with a Japanese variant form of Friedreich's ataxia. Three patients were found to have a homozygous insertion mutation of the aprataxin gene (689insT). An elder sister of a patient in this series died of cerebral hemorrhage at the age of 45, and underwent autopsy. In her cerebellar cortex, the mean density of Purkinje cells in the flocculus had predominantly decreased to 6.7% of normal controls, whereas the Purkinje cells in the other areas of the cerebellar hemisphere had decreased to 78.2%. This suggests that the cerebellar flocculus is the primary affected lesion in AOA1/EAOH, which should be associated with ocular motor apraxia. PMID- 17917454 TI - Activation of the prefrontal cortex during the wisconsin card sorting test (Keio Version) as measured by two-channel near-infrared spectroscopy in patients with traumatic brain injury. AB - To investigate brain activation in the prefrontal cortex (PFC) during the Wisconsin Card Sorting Test (Keio Version) (KWCST), we examined changes in total hemoglobin volume (THV) in 8 patients with traumatic brain injury (TBI) and 20 healthy control subjects using two-channel near-infrared spectroscopy. As a result, average THV in the right PFC during KWCST in TBI patients (-0.131 +/- 0.127) was significantly lower than in control subjects (0.016 +/- 0.135) (2 x 3 ANOVA; p < 0.05). These results demonstrated that the TBI patients had lower circulation of hemoglobin in the right PFC during the KWCST than the control subjects. PMID- 17917455 TI - Positron emission tomography facilitates diagnosis of early-onset Alzheimer's disease. AB - BACKGROUND: Clinical positron emission tomography (PET) may help in the evaluation of presenile patients with memory complaints for the presence of Alzheimer's disease (AD). METHODS: Clinical PET scans from 27 patients with clinically probable AD and early ages of onset (<65 years) were compared to PET scans from 27 age-matched controls presenting with memory complaints, but without dementia or mild cognitive impairment. RESULTS: Compared to controls, the AD patients had significant frontal, temporal and parietal hypometabolism bilaterally, and AD diagnosis correlated with left temporal and right temporoparietal hypometabolism. The sensitivity of temporoparietal hypometabolism for AD was 92.6%, the specificity 85.2%. CONCLUSION: Clinical PET imaging helps distinguish early-onset AD from patients with memory complaints not meeting criteria for dementia or mild cognitive impairment. PMID- 17917456 TI - Relationship between detectability of ischemic lesions by diffusion-weighted imaging and embolic sources in transient ischemic attacks. AB - BACKGROUND/AIMS: The aim of this study is to clarify the relationship between lesion detectability by diffusion-weighted magnetic resonance imaging (DWI) and the etiology of transient ischemic attacks (TIAs). METHODS: A retrospective study was performed on 72 patients with carotid TIAs who underwent DWI studies within 2 weeks after the last episode. RESULTS: Lesions were detected in 24 of 72 patients (33%). The detectability of lesions was 12% (3/25) in the large-artery atherosclerosis (LA) group, 57% (8/14) in the cardioembolism (CE) group, 8% (1/13) in the small-artery occlusion (SA) group, and 60% (12/20) in the other etiology or undetermined etiology (UD) group. Detectabilities in the CE group and the UD group were higher than those in the LA and SA groups. Of 24 patients with DWI-positive lesions, 17 (71%) had embolic sources in the heart; 9 were classified in the UD group because they had embolic sources both in the heart and large artery. CONCLUSION: Ischemic DWI lesions in TIAs are most likely caused by a cardioembolic mechanism. In TIA patients showing lesions on DWI, heart disease should be surveyed as the possible embolic source. PMID- 17917457 TI - Multiple sclerosis: relapses and timing of remissions. AB - BACKGROUND: Relapse is a relatively sudden worsening of neurological status in multiple sclerosis patients. The aim of this paper is to describe the frequency and timing of the subsequent clinical remission, and factors linked to a possible lack of recovery. METHODS: Two hundred and twenty-three relapses in 62 multiple sclerosis patients were examined retrospectively. Kurtzke's Expanded Disability Status Scale (EDSS) scores were calculated every 3 months from 1 year before to 1 year after. Remission was defined as improvement vs. the EDSS as scored during the relapse. Recovery was defined as return to the EDSS as scored before the relapse. RESULTS: The frequency of improvement was 78% at 3 months, 86% at 6, and lower in males (65 vs. 81%). The frequency of recovery was 55% at 3 months, 71% at 6. Kaplan-Meier curves showed stabilization after 6 months, with improvement in 90% of patients, 100% without further relapses. Relapses without recovery were only 30%; 15% excluding short-term relapses. CONCLUSIONS: Incomplete remissions do not appear to be linked to the evolution of the relapse, but to further relapses in the following months; in other words, to the short-term relapse rate. PMID- 17917458 TI - Cognitive dysfunctions in mild Parkinson's disease dementia: comparison with patients having mild Alzheimer's disease and normal controls. AB - OBJECTIVE: Although the neuropsychological differences between Alzheimer's disease (AD) and Parkinson's disease dementia (PDD) have been scrutinized in several studies, no study to date has directly compared the cognitive impairment profiles of patients with mild PDD and those with mild AD. In this study, we investigated the cognitive changes of mild PDD patients as compared with age- and education-matched normal controls and matched mild AD patients. METHODS: Thirty one patients with PDD, 24 patients with probable AD (pAD), and 50 controls were recruited prospectively. All the subjects were evaluated with a battery of detailed neuropsychological tests for the assessment of attention, memory, language, and the visuospatial and frontal executive functions. RESULTS: The PDD and pAD groups demonstrated significant impairments in all cognitive domains compared with the control group. These cognitive declines attributable to dementia were similar in the PDD and pAD patients, but they were not identical. In the subdomain analysis, the pAD group demonstrated more marked changes in delayed memory performance than did the PDD group. CONCLUSIONS: Our result showed that the cognitive changes in mild PDD patients did not differ from those in the mild pAD patients, except for delayed memory items. We concluded that it is difficult to sharply distinguish between mild PDD and mild AD. PMID- 17917459 TI - Serial assessments of delayed encephalopathy after carbon monoxide poisoning using magnetic resonance spectroscopy and diffusion tensor imaging on 3.0T system. AB - To elucidate the still unclear mechanism of delayed encephalopathy after carbon monoxide (CO) poisoning, we serially performed single-voxel proton magnetic resonance spectroscopy ((1)H-MRS) and diffusion tensor imaging (DTI) on a 3.0T system and intelligence tests in 2 patients, while they were undergoing hyperbaric oxygenation therapy. The chronological changes in (1)H-MRS- and DTI derived parameters indicated the following: (1) White matter demyelination, aerobic metabolism inhibition, and cytotoxic edema persisted for at least 3 months even after starting the hyperbaric oxygenation therapy; (2) the axonal function and structural integrity of the white matter were initially severely impaired and then gradually and partially improved for 5 months, showing changes similar to those in the scores of the intelligence tests. The results demonstrated that brain damage after CO poisoning may persist longer than expected, and that the (1)H-MRS- and DTI-derived parameters are good indicators of the clinical progress of a patient. The combination of (1)H-MRS and DTI on a 3.0T system is useful for monitoring the changes in brain damage and the clinical symptoms of patients with delayed encephalopathy after CO poisoning. PMID- 17917460 TI - Management of cerebral arteriovenous malformations associated with symptomatic congestive intracranial hypertension. AB - We report on management strategies and clinical outcomes in 4 cases of acute symptomatic congestive intracranial hypertension associated with cerebral arteriovenous malformations (AVMs). Congestion resulted from high-volume shunts exhausting the drainage capacity of the cerebral venous system in 3 patients, and from sudden venous outflow obstruction in 1 patient. Two AVMs were suggested to be surgically accessible, whereas two AVMs were deemed to be inoperable. Surgically accessible AVMs were treated with embolization followed by complete surgical resection. Inoperable AVMs were treated with partial embolization. Both AVM embolization followed by surgical resection and partial AVM embolization effectively reduced intracranial pressure and achieved sustained patient recovery. Hence, an endovascular approach may be considered to manage AVM-related congestive intracranial hypertension either in combination with surgical AVM removal, or as a palliative approach in inoperable AVMs. PMID- 17917461 TI - Association of MDR1 (C3435T) polymorphism and resistance to carbamazepine in epileptic patients from Turkey. AB - BACKGROUND AND AIMS: We investigated the prevalence of this multidrug resistance 1 gene (MDR1) polymorphism in drug-responsive versus drug-resistant epilepsy patients treated with carbamazepine (CBZ), which is a substrate of this protein. METHODS: We genotyped the C3435T variant of MDR1 in 97 patients treated with CBZ monotherapy who had been on stable doses for more than 1 month. Our control group included 174 healthy individuals. Plasma CBZ concentrations were also measured using fluorescence polarization immunoassay. RESULTS: We could not demonstrate any statistically significant relationship with the genotypes among drug resistant patients (n = 44). The frequency of the homozygous mutant (TT) genotype was 15% in drug-responsive patients, 11.3% in drug-resistant patients and 25.8% in the control group. We also did not observe any significant correlation between the presence of a specific allele and CBZ plasma level/dose index. CONCLUSION: Our study did not support any significant association between the MDR1 (C3435T) polymorphism and resistance to CBZ in epilepsy patients from Turkey. PMID- 17917462 TI - Retrocollis: classification, clinical phenotype, treatment outcomes and risk factors. AB - Retrocollis (RC) is a form of cervical dystonia (CD) that produces patterned, repetitive muscle contractions that result in neck extension. We performed a review of consecutive CD patients seen in our Division over a 15-year period. Out of 399 CD patients, 59 (14.8%) had features of RC. Pain was very frequently reported among patients ( approximately 80%). RC was frequently associated with neuroleptic exposure (20.3%) and a history of head/neck trauma (23.7%). Of patients injected with botulinum toxin type A, 24.5% reported excellent, 32.1% moderate, 16.9% mild and 24.5% no response to injections. Oral antidystonic medications had limited contribution to symptom relief. PMID- 17917463 TI - Detrended fluctuation analysis of heart rate variability in normal and growth restricted fetuses. AB - BACKGROUND: Detrended fluctuation analysis (DFA) has recently been validated as an excellent method by which to analyze heart rate variability and distinguish healthy subjects from patients with various types of the cardiac nervous system dysfunction. METHODS: One hundred and nineteen fetal heart rate (FHR) recordings obtained from healthy normal fetuses and 68 recordings obtained from small-for gestational-age (SGA) fetuses were analyzed by DFA to examine gestational and pathologic changes of the scaling exponent, alpha. RESULTS: In normal fetuses, a significant increase was observed in both the short-term (30 s) alpha(2) scaling exponents according to gestational age. The alpha(1) values of SGA fetuses were not significantly different from those of healthy normal fetuses; however, the alpha(2) values of the former group (0.955 +/- 0.152) were significantly higher than those of normal subjects (0.887 +/- 0.128; p = 0.001). CONCLUSION: The alpha(2) exponent appears to be a sensitive probe for detecting subtle, and possibly important, changes that occur in fetuses with intrauterine growth restriction, and may be helpful in the early and noninvasive detection of placental insufficiency or incipient intrauterine growth restriction. The use of DFA techniques offers great promise for understanding FHR behavior. PMID- 17917464 TI - Bladder training versus combination of propiverine with bladder training for female urinary frequency. A prospective, randomized, comparative study. AB - OBJECTIVE: Among lower urinary tract symptoms, urinary frequency is one of the prevalent symptoms in the female population. The objective of this study was to evaluate the effect of bladder training with or without pharmacotherapy in women with only a frequency symptom. METHODS: Patients were randomized to either bladder training alone (n = 23) or propiverine combined with bladder training (n = 25). All patients were treated during 3 months. RESULTS: At baseline, daytime frequency was higher in the combination therapy group than in the bladder training group (p = 0.031). No difference in other characteristics was observed in the two groups. After treatment, there was no significant difference of functional bladder capacity and average voided volume between the two groups but daytime frequency was higher in the combination therapy group than in the bladder training group (p = 0.013). However, the changes of daytime frequency, functional bladder capacity and average voided volume were not significantly different in the groups. Of total patients, 15 (65.2%) in the bladder training group and 21 (84.0%) in the combination therapy group were rated as responders (defined as improved or much improved) (p = 0.133). CONCLUSIONS: Both bladder training with or without pharmacotherapy may improve the subjective and objective frequency symptom in women with only a frequency symptom. However, we could not find that there was more symptomatic improvement when anticholinergics combined with bladder training were compared with bladder training alone. PMID- 17917466 TI - Rehabilitation in endocrine patients: a novel psychosomatic approach. AB - Long-standing endocrine disorders may imply a degree of irreversibility of the pathological process and induce highly individualized affective responses. The psychosocial impairment that is associated with incomplete remission from endocrine illness suggests the need for an innovative approach to treatment, introducing in clinical endocrinology the concept of rehabilitation, which in other fields of medicine is already established. This new proposal stems from a number of unresolved issues related to the high prevalence of psychosocial impairment in patients adequately treated for various endocrine conditions. Indeed, rehabilitation in endocrinology may be indicated in the following cases: (a) delayed recovery after appropriate treatment; (b) discrepancy between endocrine status and current functioning; (c) presence of a decline in physical and social functioning; (d) persistence of important comorbidity, with special reference to psychiatric disturbances; (e) assessment of abnormal illness behavior; (f) problems with lifestyle and risk behavior, and (g) potential role of stress in endocrine disturbances. The endocrine rehabilitation team should ideally include a trained clinical endocrinologist, a physical therapist and a psychologist, with opportunities for other specialist consultations. The goal of such service would be to ensure education, support and specific interventions, helping the patient and his/her family to achieve optimal coping with the difficulties of the recovery process. Due to its comprehensive psychosomatic characterization, this new approach would likely increase the chances of obtaining full recovery in a significant proportion of patients and has the potential of being cost-effective. PMID- 17917467 TI - Evidence for two types of bipolar depression using a dimensional approach. AB - BACKGROUND: Although there is a great heterogeneity of depressive states in bipolar patients, there is only one definition in international classifications for describing them. However, this variety seems particularly important to recognize because of the possible exacerbation of some of these bipolar depressive states by antidepressants. We aimed at assessing whether it is possible to distinguish different forms of bipolar depression using a dimensional approach. METHODS: We characterized 60 bipolar patients with a Major Depressive Episode (DSM-IV) using a new tool (MAThyS; Multidimensional Assessment of Thymic States), assessing five fundamental dimensions (emotional reactivity, cognitive speed, psychomotor function, motivation, and sensory perception) of mood states. RESULTS: A cluster analysis using the items of the dimensional scale revealed two types of depressive state: group 1 (n = 38), which had a low score, is characterized by an inhibition in all dimensions, whereas group 2 (n = 22) is characterized by an overactivation. The emotional reactivity is the most relevant dimension for discriminating these two types of depression (group 1: hyporeactivity; group 2: hyperreactivity), whereas sadness is not. CONCLUSIONS: Bipolar depressive states are not homogeneous. A dimensional approach based on emotional reactivity could be useful for discriminating the different forms of bipolar depression. Bipolar depressions may be classified as hyporeactive or hyperreactive. This classification might have therapeutic implications, because hyperreactive depression should belong to the broad spectrum of mixed states. PMID- 17917465 TI - 5-HT1B receptor subtype and aging in rat resistance vessels. AB - The effects of 5-hydroxytryptamine (5-HT), 5-carboxamidotryptamine, and sumatriptan on rat caudal arteries were examined, with the goal of finding experimental conditions useful in enhancing the 'silent' 5-HT(1B) receptor subtype. It was shown that both reserpine treatment and K(+) depolarization increased the vasoconstriction by 5-HT receptor agonists. The role of the 5 HT(2A) receptor in vasoconstriction was examined using ritanserin (50 nmol/l), a selective 5-HT(2A) antagonist, whereas that of the 'silent' 5-HT(1B) receptor was examined using SB-224289 (0.2 micromol/l), a selective 5-HT(1B) receptor antagonist. The influence of age on the 'silent' 5-HT(1B) receptor subtype was also investigated; for this, the effect of sumatriptan, a selective 5-HT(1B/1D )agonist, was tested on arterial tissues of both young and old rats which had been either K(+) depolarized or reserpine treated or both. It was found that aging strongly shifted the concentration-vasoconstriction curve generated by sumatriptan to the left, also increasing the maximum contractile response, mainly in reserpine-treated tissues. RT-PCR was used to study the expression of 5-HT(1B) and 5-HT(2A) receptors in both young and old tissues. The results support the idea that reserpine-treated and K(+)-depolarized caudal arteries from old rats can be a pharmacological model which is useful in highlighting the 'silent' 5 HT(1B) receptor subtype. PMID- 17917468 TI - Promoting mindfulness in psychotherapists in training influences the treatment results of their patients: a randomized, double-blind, controlled study. AB - BACKGROUND: All therapists direct their attention in some manner during psychotherapy. A special form of directing attention, 'mindfulness', is recommended. This study aimed to examine whether, and to what extent, promoting mindfulness in psychotherapists in training (PiT) influences the treatment results of their patients. METHODS: The therapeutic course and treatment results of 124 inpatients, who were treated for 9 weeks by 18 PiTs, were compared. The PiTs were randomly assigned to 1 of 2 groups: (i) those practicing Zen meditation (MED; n = 9 or (ii) control group, which did not perform meditation (noMED; n = 9). The results of treatment (according to the intent-to-treat principle) were examined using the Session Questionnaire for General and Differential Individual Psychotherapy (STEP), the Questionnaire of Changes in Experience and Behavior (VEV) and the Symptom Checklist (SCL-90-R). RESULTS: Compared to the noMED group (n = 61), the patients of PiTs from the MED group (n = 63) had significantly higher evaluations (according to the intent-to-treat principle) for individual therapy on 2 STEP scales, clarification and problem-solving perspectives. Their evaluations were also significantly higher for the entire therapeutic result on the VEV. Furthermore, the MED group showed greater symptom reduction than the noMED group on the Global Severity Index and 8 SCL-90-R scales, including Somatization, Insecurity in Social Contact, Obsessiveness, Anxiety, Anger/Hostility, Phobic Anxiety, Paranoid Thinking and Psychoticism. CONCLUSIONS: This study indicates that promoting mindfulness in PiTs could positively influence the therapeutic course and treatment results in their patients. PMID- 17917469 TI - Short-term psychotherapeutic interventions for somatizing patients in the general hospital: a randomized controlled study. AB - BACKGROUND: Treatment acceptance and motivation for psychotherapy of somatizing patients in the general hospital is low. METHODS: Patients (n = 91) fulfilling the criteria for somatization were randomized into an intervention group (n = 49) and a control group (n = 42). The patients in the intervention group attended 5 psychotherapeutic sessions based on the modified reattribution model. The patients in the control group received psychoeducational reading material. The primary outcomes were motivation for psychotherapy and contacting a psychotherapist after discharge. The secondary outcomes consisted of changes regarding somatoform symptoms, emotional distress and quality of life. RESULTS: Patients from the intervention group were significantly more motivated for psychotherapy (p = 0.001) than patients from the control group. At the 3-month follow-up, 42% of the patients from the intervention group had contacted a psychotherapist, compared to 20% of the patients from the control group (p = 0.045). At the 6-month follow-up, however, the ratio of patients having contacted a psychotherapist had changed to 44 and 29%, respectively, and was no longer significant. The intensity of somatoform symptoms and the anxiety symptoms decreased and mental functioning improved significantly over time for patients from both groups. CONCLUSIONS: Short-term psychotherapeutic interventions for somatizing patients in general hospitals have a moderately better effect on motivation for psychotherapy and contacting a psychotherapist than psychoeducational reading material alone. Future studies should attempt to prove the effectiveness of short-term psychoeducational interventions for somatizing patients in the general hospital. PMID- 17917470 TI - Prediction of medium-term outcome in cluster B personality disorder following residential and outpatient psychosocial treatment. AB - BACKGROUND: There is a paucity of research concerning the identification of individual characteristics predictive of outcome in the treatment of personality disorders (PDs). METHODS: In this study, we carried out a predictor analysis of a sample of 73 hospitalized patients with a primary diagnosis of cluster B PD admitted to two different psychosocial programs for PD: (a) long-term inpatient treatment, and (b) a step-down program. RESULTS: Younger age, higher Global Assessment Scale intake scores, longer length of treatment, absence of self mutilation and avoidant PDs were a significant predictor of outcome at 24-month follow-up. Self-harming patients allocated to the step-down program had higher rates of improvement compared with patients allocated to the long-term inpatient model. CONCLUSIONS: The findings may carry potential clinical implications concerning patient selection and treatment delivery for inpatient and outpatient psychosocial programs for cluster B PD. Limitations include a relatively low sample size for a regression analysis, and a larger sample of cluster B patients may be needed to ensure greater reliability of results. PMID- 17917471 TI - Increased 12-month prevalence rates of mental disorders in patients with chronic somatic diseases. AB - BACKGROUND: Although it is well established that chronic somatic diseases are significantly associated with a wide range of psychopathology, it remains unclear to what extent subjects with chronic somatic diseases are at increased risk of experiencing mental disorders. The present epidemiological study investigates age and sex-adjusted 12-month prevalence rates of mental disorders in patients with cancer, and musculoskeletal, cardiovascular and respiratory tract diseases, based on comprehensive physicians' diagnoses and compared with physically healthy probands. METHODS: Prevalence rates were calculated from two large epidemiological surveys. These studies investigated inpatients and patients from the general population with cancer (n = 174) and musculoskeletal (n = 1,416), cardiovascular (n = 915) and respiratory tract diseases (n = 453) as well as healthy controls (n = 1,083). The prevalence rates were based on the Munich Composite International Diagnostic Interview, a standardized interview for the assessment of mental disorders. RESULTS: Prevalence rates were very similar for inpatients (43.7%) and patients from the general population (42.2%). The adjusted odds ratios (OR) of patients with chronic somatic diseases were significantly elevated for mental disorders in comparison with healthy probands (OR: 2.2). Mood, anxiety and somatoform disorders were most frequent. The prevalence rates did not differ significantly between the somatic index diseases. The number of somatic diseases per patient had a higher association with mental disorders. CONCLUSIONS: There is a strong relationship between chronic somatic diseases and mental disorders. A future task is to improve the care of mental disorders in patients with chronic physical illness, specifically with multimorbid conditions. PMID- 17917472 TI - Epidemiology of life events: life events and psychiatric disorders in the Sesto Fiorentino study. AB - BACKGROUND: Although life events have been consistently reported as precipitating factors for most psychiatric disorders, there is no comprehensive investigation of the relationship between severe life events and psychiatric disorders in the general population. METHODS: This is a community-based study of psychiatric disorders among a cohort representative of adults in an Italian town. A total of 2,363 subjects out of 2,500 selected to be representative of the population living in Sesto Fiorentino, central Italy, were interviewed by their own general practitioner using the Mini International Neuropsychiatric Interview. Of the 613 subjects, 609 who resulted positive for any psychiatric disorders and 123 out of a random sample of 130 negatives were re-interviewed by the psychiatrists using the Florence Psychiatric Interview. The Florence Psychiatric Interview was used to explore each distinct psychiatric episode. Life events were recorded in detail by a specific interview. RESULTS: During the year prior to the onset of the first psychiatric disorder, 35.8% of cases suffered from at least a severe event, compared with 12.2% of non-cases during a comparable period (OR = 4.0, 95% CI = 2.3-7.1). The excess of life events occurred for almost all the diagnostic categories. The same results were reproduced even when only the 'independent' life events were considered. The distribution of the events through the 12 months taken into account showed an even distribution of events among non-cases, whereas there was a clear accumulation in the last 3 months prior to the onset of the pathology in the cases. CONCLUSIONS: Life stress is one of the main precipitating factors of psychopathology. PMID- 17917473 TI - Genetic and environmental factors in alexithymia: a population-based study of 8,785 Danish twin pairs. AB - BACKGROUND: The role of genetic and environmental factors for developing alexithymia is still unclear, and the aim of this study was to examine these factors in a large population-based sample of twins. METHODS: The Toronto Alexithymia Scale-20 (TAS-20) was included in a mail survey of 46,418 individuals born between 1931 and 1982 and registered with the Danish Twin Registry. The response rate was 75.3%. A total of 8,785 twin pairs, where both cotwins had completed all items of the TAS-20, were selected for this study. Analyses were conducted for total TAS-20 scores and the subscales of (1) difficulties in identifying feelings, (2) difficulties in describing feelings, and (3) externally oriented thinking. The phenotypes were analyzed both as categorical and continuous data. RESULTS: All measures of similarity suggested that genetic factors added to all facets of alexithymia. Structural equation modeling of the noncategorical data, an ACE model including additive genetic, shared environmental and nonshared environmental effects, provided the best fit for all three facets of alexithymia as well as total alexithymia scores, with heritabilities of 30-33% and the remaining variance being explained by shared (12 20%) and nonshared environmental effects (50-56%). CONCLUSION: The results from this large population-based sample suggest that genetic factors have a noticeable and similar impact on all facets of alexithymia. While the results suggested a moderate influence of shared environmental factors, our results are in concordance with the general finding that environmental influences on most psychological traits are primarily of the nonshared rather than the shared type. PMID- 17917474 TI - Chemotherapy-induced nausea, vomiting, and fatigue--the role of individual differences related to sensory perception and autonomic reactivity. AB - BACKGROUND: In spite of antiemetics, postchemotherapy side effects continue to be common and may affect compliance to cancer treatment. Among the known factors associated with increased symptom severity are: younger age, treatment toxicity, expected severity, and distress, but little is still known about the role of other factors. The aim of our study was to investigate the role of individual differences related to sensory perception for posttreatment side effects. METHODS: Hundred and twenty-five women receiving adjuvant chemotherapy for breast cancer completed measures of absorption, autonomic perception, somatosensory amplification, trait anxiety, and expected severity at baseline. Pretreatment distress and posttreatment nausea, vomiting, and fatigue were assessed at the 1st, 4th, 6th and last cycles of chemotherapy. RESULTS: While univariate analyses showed several factors to be associated with side effects, only absorption and pretreatment distress remained independent predictors of nausea and fatigue when controlling for the remaining factors. Posttreatment vomiting was only predicted by expected severity of vomiting. CONCLUSION: Chemotherapy-induced side effects are related to increased autonomic nervous system activity, and absorption has been associated with increased autonomic nervous system reactivity to stress. The results suggest that individuals with high absorption may be at greater risk for developing side effects. Improved precision in identifying patients at risk of experiencing more severe side effects after cancer treatment will increase the ability to target treatments aimed at reducing these side effects. PMID- 17917475 TI - The startle reflex in alcohol-dependent patients: changes after cognitive behavioral therapy and predictive validity for drinking behavior. A pilot study. AB - BACKGROUND: Previous studies demonstrated an attenuation of the affect-modulated startle reflex when alcohol-dependent patients were viewing alcohol-associated pictures. This indicates an appetitive valence of these stimuli. We used the affect-modulated startle reflex to assess the effects of behavioral treatment on the emotional processing of alcohol-associated stimuli. Further, we examined whether the affect-modulated startle reflex is a predictor of treatment success. METHODS: Forty-three alcohol-dependent patients (21 females, mean age 45.67 years, SD 9.45) were recruited consecutively from an inpatient alcohol detoxification facility where patients attended a 3-week detoxification program including cognitive-behavioral treatment to successfully handle high-risk situations. The eye blink component of the affect-modulated startle response, self-reported cue-induced craving and skin conductance responses to alcohol associated and control slides were assessed before and after treatment. Changes were analyzed using repeated measures analysis of variance. Drinking behavior was assessed in the 6 months following treatment, and a regression analysis was performed to evaluate the predictive validity of the affect-modulated startle response for drinking behavior. RESULTS: Drinking behavior as well as craving and skin conductance responses decreased significantly over time. The pattern of the affective modulation of the startle reflex was not altered over time. However, startle modulation and relapse were related, and within the group of relapsers, startle modulation was a significant predictor of drinking behavior. CONCLUSIONS: Our results suggest that the modulation of the startle reflex may reflect more enduring and permanent processes of emotional responding to alcohol-related cues than autonomic arousal and self-reported craving, and that startle modulation by alcohol-associated cues may be a better predictor of drinking behavior for relapsers than other measures. Further studies including a control condition are necessary to validate these findings. PMID- 17917476 TI - Psychosocial factors associated with preterm uterine contractions. AB - BACKGROUND: The aim of our study was to evaluate how sociodemographic factors, psychosocial adaptation to pregnancy and well-being levels are associated with the onset of preterm uterine contractions allowing symptomatic preterm labor. METHODS: In a prospective case-control design, 51 consecutive women admitted for threatened preterm labor were enrolled. The patients received standard care. The day before discharge, once contractions had been stopped, the patients were administered 2 questionnaires: the Prenatal Self-Evaluation Questionnaire of Lederman and the Psychological Well-Being Scales. Controls were enrolled among asymptomatic, healthy women attending routine prenatal care. They were matched for parity and gestational age. RESULTS: Gestational age at inclusion ranged from 25 to 34 weeks. Fourteen cases and 4 controls delivered preterm. Cases were less educated than controls, showed a lower acceptance of pregnancy and worse relationship with others, namely with the husband, compared to controls. They also displayed a reduced environmental mastery. CONCLUSION: Having a low education, poor relationship with others, including the husband, and impaired coping skills appeared to be independent risk factors for the development of symptomatic preterm labor in urbanized women. PMID- 17917477 TI - Evaluation and treatment of sleep complaints: patients' subjective responses. AB - BACKGROUND: This study set out to evaluate whether patients' expectations affected their responses to treatment of a sleep disorder. METHODS: Fifty consecutive patients attending a sleep clinic for the first time and who had different diagnoses of sleep disorders were included in this study. The patients completed a set of 6 questionnaires on 7 occasions as they progressed through the process of a sleep clinic referral, assessment and treatment. Self-report questionnaires were used to assess their sleepiness, fatigue and alertness. Predictor measures included mental health and individual items assessing expectation regarding the seriousness of the sleep problem. A battery of questions dealt with mental health issues, patients' expectation and their commitment to the sleep investigation and treatment process. Each patient's responses were examined over a period of 6 months. RESULTS: Immediately following their interview with the sleep consultant, the patients' concern regarding their sleep problem was higher than the first measures obtained at baseline. Compared to prior to their first consultation with a sleep specialist, while on treatment they directed more attention to their sleep problem when problems associated with fatigue were reduced. Although there was a lessening in fatigue with treatment, subjectively rated sleepiness (Epworth Sleepiness Scale) did not vary over the study. Contact with a sleep center and the diagnosis and treatment of individual sleep problems also resulted in improved satisfaction with life. CONCLUSIONS: Contact with and treatment at a sleep clinic was found to be beneficial. The fatigue levels were reduced and the patients had greater life satisfaction regardless of the diagnosis and treatment of their sleep disorder, although subjective sleepiness did not change. Patient expectations were not critical in determining the outcome of the sleep clinic assessment. PMID- 17917478 TI - Scientific study of the dissociative disorders. PMID- 17917480 TI - Neural correlates of psychodynamic psychotherapy in borderline disorders--a pilot investigation. PMID- 17917481 TI - [Quality of life and its associated factors among some elderly residents using a hall for the aged in a community]. AB - OBJECTIVES: This study was performed to investigate the level of quality of life (QOL) and to identify its related factors among the elderly using a hall for the aged. METHODS: The study subjects were 350 elderly people living in Nam Gu, Gwangju City. They were interviewed by questionnaire to collect information on social-demographic characteristics, health behaviors, health-related characteristics and QOL. We used statistical analysis methods such as T-test, ANOVA and multiple regression analysis to find which factors affected QOL. RESULTS: The characteristics relating to physical functions were as follows: presence of spouse, education, alcohol drinking, teeth condition, arthritis, frequency of visiting the hall for the aged, and depression. Age, arthritis and depression were significantly related to role limitation due to physical problems. General health was related to gender, age, and depression, and vitality was related to education, frequency of visiting the hall, and depression. Role limitation due to emotional problems was related to age and depression, and mental health was related to age, hypertension, arthritis and depression. CONCLUSIONS: This study showed QOL of the elderly was significantly influenced by socio-demographic factors, and physical or emotional conditions. To improve QOL of the elderly, we need to develop programs to promote health and to manage chronic diseases of the elderly. PMID- 17917482 TI - [Disparities in participation in health examination by socio-economic position among adult Seoul residents]. AB - OBJECTIVES: To determine the disparity in the rate people undergo health examinations according to socioeconomic position (SEP) and the changes in this disparity with time. METHODS: Seoul citizens' health profile data from 1997 to 2005 were analyzed. The study subjects were 40 years old and over, and the total number of subjects was 6,601 in 1997, 8,994 in 2001, and 8,819 in 2005. Those aged 60 years and over were eliminated from the analysis of subjects' occupation. We used education, family income and occupation as indicators of SEP. The age standardized health examination attendance rate for each year was calculated according to the education, family income and occupation. The odds ratios (ORs) from multiple logistic regressions were adjusted for age. RESULTS: The disparity in the rate of attendance according to the SEP decreased from 1997 to 2005 but still existed. Even though the disparities among the subgroups according to education, family income and occupation were not that high, the disparity between the group with the highest SEP and the other groups was considerable. CONCLUSIONS: Our findings suggest that unequal access to health examination services according to socioeconomic position still exists. This disparity has decreased recently but the disparity according to level of education was the greatest. PMID- 17917483 TI - [Relationship between job stress contents, psychosocial factors and mental health status among university hospital nurses in Korea]. AB - OBJECTIVES: The present study was intended to assess the mental health of nurses working for university hospitals and to establish which factors determine their mental health. METHODS: Self-administered questionnaires were given to 1,486 nurses employed in six participating hospitals located in Daejeon City and Chungnam Province between July 1st and August 31st, 2006. The questionnaire items included sociodemographic, job-related, and psychosocial factors, with job stress factors (JCQ) as independent variables and indices of mental health status (PWI, SDS and MFS) as dependent variables. For statistical analysis, the Chi-square test was used for categorical variables, with hierarchical multiple regression used for determining the factors effecting mental health. The influence of psychosocial and job-related factors on mental health status was assessed by covariance structure analysis. The statistical significance was set at p<0.05. RESULTS: The factors influencing mental health status among subject nurses included sociodemographic characteristics such as age, number of hours of sleep, number of hours of leisure, and subjective health status; job-related characteristics such as status, job satisfaction, job suitability, stresses such as demands of the job, autonomy, and coworker support; and psychosocial factors such as self-esteem, locus of control and type A behavior patterns. Psychosocial factors had the greatest impact on mental health. Covariance structure analysis determined that psychosocial factors affected job stress levels and mental health status, and that the lower job stress levels were associated with better mental health. CONCLUSIONS: Based on the study results, improvement of mental health status among nurses requires the development and application of programs to manage job stress factors and/or psychosocial factors as well as sociodemographic and job-related characteristics. PMID- 17917484 TI - [Environmental health surveillance of low birth weight in Seoul using air monitoring and birth data]. AB - OBJECTIVES: The principal objective of this study was to determine the relationship between maternal exposure to air pollution and low birth weight and to propose a possible environmental health surveillance system for low birth weight. METHODS: We acquired air monitoring data for Seoul from the Ministry of Environment, the meteorological data from the Korean Meteorological Administration, the exposure assessments from the National Institute of Environmental Research, and the birth data from the Korean National Statistical Office between January 1, 2002 and December 31, 2003. The final birth data were limited to singletons within 37-44 weeks of gestational age. We defined the Low Birth Weight (LBW) group as infants with birth weights of less than 2500g and calculated the annual LBW rate by district. The air monitoring data were measured for CO, SO(2), NO(2), and PM(10) concentrations at 27 monitoring stations in Seoul. We utilized two models to evaluate the effects of air pollution on low birth weight: the first was the relationship between the annual concentration of air pollution and low birth weight (LBW) by individual and district, and the second involved a GIS exposure model constructed by Arc View 3.1. RESULTS: LBW risk (by Gu, or district) was significantly increased to 1.113(95% CI=1.111 1.116) for CO, 1.004 (95% CI=1.003-1.005) for NO(2), 1.202(95% CI=1.199-1.206) for SO(2), and 1.077(95% CI=1.075-1.078) for PM(10) with each interquartile range change. Personal LBW risk was significantly increased to 1.081(95% CI=1.002 1.166) for CO, 1.145(95% CI=1.036-1.267) for SO(2), and 1.053(95% CI=1.002-1.108) for PM(10) with each interquartile range change. Personal LBW risk was increased to 1.003(95% CI=0.954-1.055) for NO(2), but this was not statistically significant. The air pollution concentrations predicted by GIS positively correlated with the numbers of low birth weights, particularly in highly polluted regions. CONCLUSIONS: Environmental health surveillance is a systemic, ongoing collection effort including the analysis of data correlated with environmentally associated diseases and exposures. In addition, environmental health surveillance allows for a timely dissemination of information to those who require that information in order to take effective action. GIS modeling is crucially important for this purpose, and thus we attempted to develop a GIS-based environmental surveillance system for low birth weight. PMID- 17917485 TI - [The effect of cigarette price on smoking behavior in Korea]. AB - OBJECTIVES: To determine the impact of cigarette prices on the decision to initiate and quit smoking by taking into account the interdependence of smoking and other behavioral risk factors. METHODS: The study population consisted of 3,000 male Koreans aged > or =20. A survey by telephone interview was undertaken to collect information on cigarette price, smoking and other behavioral risk factors. A two-part model was used to examine separately the effect of price on the decision to be a smoker, and on the amount of cigarettes smoked. RESULTS: The overall price elasticity of cigarettes was estimated at -0.66, with a price elasticity of -0.02 for smoking participation and -0.64 for the amount of cigarettes consumed by smokers. The inclusion of other behavioral risk factors reduced the estimated price elasticity for smoking participation substantially, but had no effect on the conditional price elasticity for the quantity of cigarettes smoked. CONCLUSIONS: From the public health and financial perspectives, an increase in cigarette price would significantly reduce smoking prevalence as well as cigarette consumption by smokers in Korea. PMID- 17917486 TI - [Differential effects of family income on self-rated health by age: analysis of Seoul citizens health indicators survey 2001, 2005]. AB - OBJECTIVES: This study was conducted in order to determine how the association between socioeconomic position(SEP) and health status changes with age among Seoul residents aged 25 and over. METHODS: We utilized the 2001 and 2005 Seoul Citizens Health Indicators Surveys. We used self-rated 'poor' health status as an outcome variable, and family income as an indicator of SEP. In order to characterize the differential effects of socioeconomic position on health by age, we conducted separate multivariate analyses by 10-year age groups, controlling for sociodemographic covariates. In order to assess the relative health inequality across socioeconomic groups, we estimated the Relative Index of Inequality (RII). RESULTS: The risk of 'poor health' is significantly high in low family income groups, and this increased risk is seen at all ages. However, the magnitude of relative socioeconomic inequality in health, as measured by the odds ratio and RII, is not identical across age groups. The difference in health across income groups is small in early adulthood (ages 25-34), but increases with age until relatively late in life (ages 35-64). It then decreases among the elderly population (ages more than 65). When the RII reported in 2005 is compared to that reported in 2001, RII can be seen to have increased across all ages, with the exception of individuals aged 25-34. CONCLUSIONS: The magnitude of health inequality is the greatest during mid- to late adulthood (ages 45-64). In addition, health inequalities have worsened between 2001 and 2005 across all age groups after economic crisis. PMID- 17917487 TI - [Health inequity among waged workers by employment status]. AB - OBJECTIVES: The purpose of this study was to examine the relationship between the differences in employment status and self assessed health in Korea. METHODS: We analyzed 4 year follow-up data generated by the Korean Labor and Income Panel Study(KLIPS), which was conducted on 1,207 men and 582 women who had undergone a change in employment status. The study subjects were placed into 1 of the following 4 groups based on their employment history; Non-precarious workers, Precarious to non-precarious workers, Non-precarious to precarious workers and Precarious workers. Logistic regression was then used to examine the relationship between the changes in employment status and self assessed health. RESULTS: When males were considered, self assessed health was better among the precarious to non-precarious workers (OR 1.58, 95% CI=1.57-1.60) and the precarious workers (OR 1.29, 95% CI=1.28-1.30) than in the non-precarious workers, after adjusting for age, socioeconomic status (education level, occupational class, marital status, average equivalent household income and average number of hours worked per week), health behavior (smoking, drinking and exercise) and medical service access (regular medical examination, have chronic disease or hospitalized within 1 year). When female workers were considered, the precarious to non-precarious workers (OR 1.89, 95% CI=1.86-1.92), non-precarious to precarious workers(OR 1.24, 95% CI=1.23-1.26) and precarious workers (OR 1.27, 95% CI=1.25-1.28) all reported poorer health than the non-precarious workers after adjusting for the aforementioned factors. CONCLUSIONS: This study showed that changes in employment status were associated with differences in self assessed health among men and women. Specifically, the results of this study showed that a corresponding positive outcome based on self assessed health was greater for employees that changed from precarious to non-precarious jobs and for male employees with precarious jobs, whereas female employees with non-precarious jobs had higher self assessed health. However, additional longitudinal studies on the health effects of employment status should be conducted. PMID- 17917488 TI - [The relationship between metabolic syndrome and Korean cardiocerebrovascular risk assessment: for male researchers in a workplace]. AB - OBJECTIVES: The prevalence of metabolic syndrome has recently increased. Payments from the Korea Labor Welfare Corporation for compensation for mortality in workers caused by cardiovascular and cerebrovascular diseases have also increased in Korea in recent years. The association of metabolic syndrome and cardiocerebrovascular disease has been investigated by several researchers in recent studies. This study was conducted in an attempt to characterize the relationship between metabolic syndrome and Korean cardiocerebrovascular disease risk assessment, and to provide basic data to group health practices for the prevention of cardiocerebrovascular disease. METHODS: Health examinations were previously conducted for 1526 male researchers at a private laboratory. The prevalence by age and the odds ratio of metabolic syndrome scores into the "cardiocerebrovascular risk group" (sum of low, intermediate, and high risk groups) of the Korean cardiocerebrovascular disease risk assessment were assessed, in an effort to elucidate the associations between metabolic syndrome and cardiocerebrovascular disease risk assessment. RESULTS: The prevalence of metabolic syndrome and inclusion in the cardiocerebrovascular risk group was 11.7% and 22.1% respectively. The severity of metabolic syndrome and cardiocerebrovascular risk assessment showed that individuals in their 40's and 50's were at higher risk than those in their 30's (p<0.001). The age-adjusted odds ratio of metabolic syndrome to cardiocerebrovascular risk group inclusion was 5.6. CONCLUSIONS: An active prevention program for cardiocerebrovascular disease needs to begin in the 40's, as the prevalence of metabolic syndrome and the risk group of cardiocerebrovascular risk assessment peak in the 40's age group. The odds ratio between metabolic syndrome and the cardiocerebrovascular risk group was high, which indicates that metabolic syndrome scores should be utilized as guidelines during the consultation and behavioral modification program for the workplace prevention of cardiocerebrovascular diseases in group health practices. PMID- 17917489 TI - [Socioeconomic inequalities in preventive services among the elderly: results from medical checkup, cancer check, and BP check]. AB - OBJECTIVES: Due to the assumptions of homogeneity as well as challenges in the socioeconomic position of the elderly, they have been relatively neglected in studies of health inequalities. Therefore, this study was conducted to investigate the social inequalities in preventive services among elderly men and women. METHODS: Data were obtained from a nationally representative sample of 342 men and 525 women aged 65 and over collected during the 2001 National Health and Nutrition Examination Survey. Age adjusted proportions and logistic regression were used to identify the social patterning of preventive services among elderly Koreans using various social position indicators. RESULTS: The findings of this study generally supported the presence of social gradients in preventive services among the Korean elderly. The likelihood of using the service becomes progressively higher with social position. Educational level, income, and self rated living status were significantly associated with increased medical checkups and cancer checks. In addition, logistic regression detected educational inequalities only among older women receiving BP checks. After being stratified based on health status and chronic disease status, social disparities still existed when educational level and self-rated living status were considered. Among unhealthy individuals, place of residence was observed as a barrier to medical checkups. CONCLUSIONS: This study demonstrated strong and consistent associations between socioeconomic position and preventive services among the elderly in Korea. The results indicate that public health strategies should be developed to reduce the barriers to preventive services encountered by the elderly. PMID- 17917490 TI - Impact of multiple cardiovascular risk factors on the carotid intima-media thickness in young adults: the Kangwha Study. AB - OBJECTIVES: Although risk factors for coronary artery disease are also associated with increased carotid intima-media thickness (IMT), there is little information available on the asymptomatic, young adult population. We examined the association between multiple cardiovascular risk factors and the common carotid IMT in 280 young Korean adults. METHODS: The data used for this study was obtained from 280 subjects (130 men and 150 women) aged 25 years who participated in the Kangwha Study follow-up examination in 2005. We measured cardiovascular risk factors, including anthropometrics, blood pressure, blood chemistry, carotid ultrasonography, and reviewed questionnaires on health behaviors. Risk factors were defined as values above the sex-specific 75th percentile of systolic blood pressure, body mass index, total cholesterol/high-density lipoprotein cholesterol ratio, fasting blood glucose and smoking status. RESULTS: The mean carotid IMT +/ standard deviation observed was 0.683 +/- 0.079 mm in men and 0.678 +/- 0.067 mm in women (p=0.567) and the evidence of plaque was not observed in any individuals. Mean carotid IMT increased with an increasing number of risk factors(p for trend <0.001) and carotid IMT values were 0.665 mm, 0.674 mm, 0.686 mm, 0.702 mm, and 0.748 mm for 0, 1, 2, 3, and 4 to 5 risk factors, respectively. The odds ratio for having the top quartile carotid IMT in men with 3 or more risk factors versus 0-2 risk factors was 5.09 (95% CI, 2.05-12.64). CONCLUSIONS: Current findings indicate the need for prevention and control of cardiovascular risk factors in young adults and more focus on those with multiple cardiovascular risk factors. PMID- 17917491 TI - T-wave amplitude attenuation/augmentation in patients with changing edematous states: implications for patients with congestive heart failure. AB - Since peripheral edema impacts the entire electrocardiographic curve, it was hypothesized that it would also affect T waves. The amplitude of T waves were measured in all electrocardiographic leads and a sum (SigmaT) was calculated in 28 patients with and 28 patients without peripheral edema (controls). For patients with peripheral edema, SigmaT on admission was 21.9+/-10.6 mm and SigmaT at peak weight was 8.3+/-6.3 mm (P=.0005). For patients with peripheral edema who subsequently lost weight, SigmaT at peak weight was 7.2+/-6.1 mm and SigmaT at the lowest weight was 14.1+/-12.2 (P=.006). For controls, SigmaT from admission and SigmaT from discharge were 24.4+/-16.9 mm and 24.7+/-15.7 mm (P=.82), respectively. Percent change (Delta%SigmaT) from admission to peak weight correlated with Delta% in weight (r=0.58; P=.001) and Delta% in the sum of QRS complexes (SigmaQRS) (r=0.71; P=.00005). Delta%SigmaT from peak weight to the lowest weight correlated with the corresponding Delta%SigmaQRS (r=0.65; P=.02). Changes in T waves with development and alleviation of peripheral edema mirror the changes shown by the QRS complexes and may be useful in the treatment of patients with congestive heart failure or other edematous states. PMID- 17917492 TI - Seasonal variation and trends in heart failure morbidity and mortality in a South American community hospital. AB - Circannual variation in cardiovascular events is well-known. Seasonal variation (SV) in heart failure (HF) has been described in the Northern Hemisphere, but there is scarcity of data in South America. The goals of the present study were to (1) describe the SV of admissions and deaths for HF, and (2) describe trends in HF morbidity and mortality in a community hospital in Argentina from 1992 to 1999. The study sample included 6369 admissions. During this period, the number of HF admissions rose by 188% and the rate HF admissions/all admissions increased from 3.28% to 7.84%. In-hospital mortality decreased from 21% to 13%. HF admissions followed a seasonal pattern with a winter-spring predominance. Male and very old patients were the subgroups with the highest SV. The authors identified clear SV in HF deaths and admissions, which raises a different hypothesis about the rationale of HF admissions and provides information for the organization of care and resource allocation. PMID- 17917493 TI - Advanced heart failure: prognosis, uncertainty, and decision making. AB - Heart failure is a serious clinical management challenge for both patients and primary care physicians. The authors studied the perceptions and practices of internal medicine residents and faculty at an academic medical center in the Southeast to guide design of strategies to improve heart failure care. Data were collected via a self-administered survey. Eighty-nine faculty and resident physicians in general internal medicine and geriatrics participated (74% response rate). Items measured perceived skills and barriers, adherence to guidelines, and physician understanding of patient prognosis. Case studies explored practice approaches. Clinical knowledge and related scales were generally good and comparable between physician groups. Palliative care and prognostic skills were self-rated with wide variance. Physicians rated patient noncompliance and low lifestyle change motivation as major barriers. Given the complexities of caring for elderly persons with heart failure and comorbid conditions, there are significant opportunities for improving physician skills in decision making, patient-centered counseling, and palliative care. PMID- 17917495 TI - Elements of nonpharmacologic interventions that prevent progression of heart failure: a meta-analysis. AB - This review examined whether nonpharmacologic treatment was associated with reductions in all-cause mortality and heart failure (HF) hospitalizations and investigated the effects of face-to-face contact and longer treatment duration on these outcomes. MEDLINE and PsycINFO databases were searched through June 2006 and bibliographies of potential articles were hand-searched. Nonpharmacologic treatment was associated with significantly lower odds of HF hospitalizations (odds ratio [OR], 0.41; 95% confidence interval [CI], 0.30-0.56) and death (OR, 0.69; 95% CI, 0.56-0.85) compared with control treatment. Face-to-face contact was associated with significantly lower odds of HF hospitalization (OR, 0.42; 95% CI, 0.22-0.81; P<.05) and death (OR, 0.63; 95% CI, 0.44-0.91; P<.05) as compared with control treatment. Longer treatment duration (>or=12 months) was associated with a 65% reduction in the rate of HF hospitalizations and a 36% reduction in death rate. Nonpharmacologic treatment featuring face-to-face contact is particularly effective in reducing HF hospitalization and all-cause mortality rates. PMID- 17917494 TI - Markers of cardiac collagen turnover are similar in patients with mild and more severe symptoms of heart failure. AB - Cardiac fibrosis plays an important role in the pathophysiology of heart failure. The authors sought to determine whether biomarkers of cardiac fibrosis for milder clinical degrees of heart failure are comparable to those of more advanced disease. Procollagen types I and III amino-terminal peptides (PINP and PIIINP) and type I collagen telopeptide (ICTP) were compared between aldosterone antagonistnaive patients with heart failure and New York Heart Association class I or II (n=22/23) and class III or IV (n=42/3) symptoms. Median (interquartile) range concentrations of PINP (63.3 [44.2-88.8] vs 48.6 [37.8-74.9] microg/L), ICTP (7.0 [5.4-16.8] vs 6.5 [4.7-12.7] microg/L), and PIIINP (4.7 [3.2-7.0] vs 4.7 [2.9-7.3] microg/L) were comparable between patients with mild and moderate to severe disease, respectively. These data suggest that patients with mild heart failure may have similar degrees of cardiac fibrosis to patients with more severe disease and support the examination of antifibrotic therapy, including aldosterone antagonists, in milder degrees of heart failure. PMID- 17917496 TI - Erythropoietin in chronic heart failure. AB - In patients with chronic heart failure (CHF), anemia is common and is associated with adverse outcome. Correction of anemia by erythropoiesis-stimulating proteins would thus seem attractive. Endogenous erythropoietin (Epo) levels are increased in CHF and are associated with severity of the disease and with increased mortality. Furthermore, Epo levels poorly correlate with hemoglobin levels, suggesting that elevated Epo levels are not only driven by anemia, but by the condition of CHF as well. Several experimental studies have demonstrated ancillary cardioprotective effects of the recombinant form of Epo, including reduced apoptosis and increased neovascularization. Three early, small studies and 3 subsequent phase 2 trials found that erythropoiesis-stimulating proteins in anemic CHF patients were safe overall and potentially beneficial. Currently, a large phase 3, randomized, clinical trial is ongoing that evaluates the effects of darbepoetin alpha on morbidity and mortality in CHF. PMID- 17917497 TI - Rosiglitazone and heart failure: the controversy and clinical implications. AB - In this issue of Congestive Heart Failure, the editor of this department addresses a topic on which there is conflicting data. Dr Tepper selected 2 of the most relevant abstracts for review. His comments on the conflicting data-and the polarized viewpoints among physicians that often result-provide insights on the clinical implications of the controversy. PMID- 17917498 TI - Historical vignettes in heart failure. PMID- 17917499 TI - Anticoagulation in severe heart failure. PMID- 17917500 TI - Would preemptive therapy in childhood prevent cardiovascular disease? PMID- 17917502 TI - A cost-effectiveness analysis of angiotensin-converting enzyme inhibitors and angiotensin receptor blockers in diabetic nephropathy. AB - The aim of this study was to estimate the cost-effectiveness of renin-angiotensin aldosterone system blockers in patients with diabetic nephropathy. A cost effectiveness analysis was performed based on a meta-analysis of studies investigating the effect of angiotensin-converting enzyme (ACE) inhibitors and angiotensin receptor blockers (ARBs) as part of a treatment regimen on the incidence of end-stage renal disease (ESRD) in patients with diabetic nephropathy. The primary outcome was the cost to prevent 1 patient from developing ESRD. Cost analysis was performed from a third-party payer perspective in 2006 US dollars. As part of a treatment regimen, ARBs significantly reduced the incidence of ESRD and doubling of serum creatinine concentration (P<.05) but not total mortality. The cost to prevent 1 patient from developing ESRD was $31,729 (95% confidence interval, $19,443-$85,442; P<.01), $189,190 (P=.13) and $51,585 (P=.068) for patients receiving ARBs, ACE inhibitors, or either of them, respectively. This study demonstrates that blocking the RAAS, which delays the progression to ESRD, appears to be cost-effective. The current analysis favors ARBs in terms of cost-effectiveness. PMID- 17917501 TI - Antihypertensive efficacy of the oral direct renin inhibitor aliskiren as add-on therapy in patients not responding to amlodipine monotherapy. AB - This study investigated the addition of the direct renin inhibitor aliskiren to amlodipine in patients with mild to moderate hypertension that was inadequately controlled with amlodipine alone. Following once-daily treatment with amlodipine 5 mg for 4 weeks, patients whose hypertension responded inadequately to therapy (mean sitting diastolic blood pressure [DBP] 90-109 mm Hg) (n=545) were randomized to 6 weeks of double-blind treatment with amlodipine 5 mg plus aliskiren 150 mg, amlodipine 5 mg, or amlodipine 10 mg. At the study's end, mean systolic blood pressure and DBP reductions with the combination of aliskiren 150 mg and amlodipine 5 mg (11.0/8.5 mm Hg) were significantly greater (P<.0001) than with amlodipine 5 mg (5.0/4.8 mm Hg)--the comparator group--but similar to amlodipine 10 mg (9.6/8.0 mm Hg). All treatments were well tolerated. Edema occurred more frequently with amlodipine 10 mg (11.2%) than with combination therapy (2.1%) or amlodipine 5 mg (3.4%). In conclusion, aliskiren 150 mg plus amlodipine 5 mg shows similar but not better blood pressure-lowering efficacy when compared with amlodipine 10 mg in patients not completely responsive to amlodipine 5 mg; less edema was noted with combination therapy. PMID- 17917503 TI - Efficacy and safety of darusentan in patients with resistant hypertension: results from a randomized, double-blind, placebo-controlled dose-ranging study. AB - In this phase 2, randomized, double-blind, placebo-controlled forced dose titration study, 115 patients with resistant hypertension, receiving background therapy with >/=3 antihypertensive medications including a diuretic at full doses, were randomized 2:1 to increasing doses of darusentan (10, 50, 100, 150, and 300 mg), a selective endothelin receptor antagonist, or matching placebo once daily for 10 weeks. Darusentan treatment decreased mean systolic and diastolic blood pressure levels in a dose-dependent fashion compared with placebo; the largest reductions were observed at week 10 (300-mg dose) (systolic, -11.5+/-3.1 mm Hg [P=.015;] diastolic, -6.3+/-2.0 mm Hg [P=.002]). Darusentan (300 mg) also decreased mean 24-hour, daytime, and nighttime ambulatory blood pressures from baseline to week 10. Darusentan was generally well tolerated; mild to moderate edema and headache were the most common adverse events. This study demonstrates a clinical benefit from a new class of antihypertensive agent in patients classified as resistant by the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure guidelines. PMID- 17917504 TI - Aldosterone escape with diuretic or angiotensin-converting enzyme inhibitor/angiotensin II receptor blocker combination therapy in patients with mild to moderate hypertension. AB - Renin-angiotensin-aldosterone system (RAAS) hyperactivity is implicated in the development of hypertension and progressive damage in target organs. Chronic inhibition of the RAAS or use of thiazide-type diuretics may trigger an aldoster one escape. The aim of this study was to assess this phenomenon in hypertensive patients treated with thiazide-type diuretics (hydrochlorothiazide [HCTZ]) or single or double blockade of the RAAS (irbesartan [IRBE], quinapril [QUIN], and IRBE+QUIN). Blood pressure levels were obtained by 24-hour ambulatory blood pressure monitoring. Plasma renin activity and aldosterone levels were determined by immunoradiometric assay. Blood pressure level was normalized in the 4 treatment groups; the HCTZ and IRBE+QUIN groups showed an increased plasma aldosterone level after 12 weeks (9.1+/-2.2 to 14.1+/-1.4 and 6.9+/-1.9 to 12.9+/ 2.3 ng/dL, respectively; P<.05), whereas plasma renin activity was increased only in the HCTZ group (0.9+/-0.2-1.7+/-0.2 ng/mL/h; P<.05). The increase in plasma aldosterone level after 12 weeks of HCTZ and IRBE+QUIN therapy suggests early aldosterone escape. PMID- 17917505 TI - Differential impacts of adiponectin on low-grade albuminuria between obese and nonobese persons without diabetes. AB - The goal of this study was to examine the association between adiponectin and low grade albuminuria in nondiabetic persons and whether it differed between obese and nonobese individuals. Urinary albumin excretion (UAE) was assessed by overnight collection in 157 participants. Overall, as anticipated, 24-hour systolic blood pressure and fasting glucose levels were independent determinants of UAE (beta=0.254 and 0.176, respectively, P<.05). In obese persons (body mass index [BMI] >/=25 kg/m(2); n=63), adiponectin value was a significant negative determinant of UAE (beta=-0.256, P<.05) independent of blood pressure and glucose levels, whereas in nonobese participants (n=94) it was not. In an analysis of BMI and the median value of adiponectin (9.9 microg/mL), UAE was found to be significantly higher in obese persons with low adiponectin levels than in obese persons with high adiponectin levels (9.20 vs 5.11 microg/min; P<.05), even after adjustment for age, sex, blood pressure level, and glucose level. There was an inverse association between adiponectin and low-grade albuminuria in obese nondiabetic persons. PMID- 17917506 TI - Combination therapy with an angiotensin-converting enzyme inhibitor and a calcium channel blocker. AB - More than 1 medication is required in many hypertensive patients to reach blood pressure (BP) goals, and initial treatment with 2 agents has been recommended for patients whose BP level is >20/10 mm Hg above target. Diuretics reduce BP levels and the incidence of target organ complications and together with angiotensin converting enzyme inhibitors (ACEIs) or angiotensin receptor blockers, which are recommended in patients with comorbid cardiovascular disease, nephropathy, or diabetes, are effective antihypertensive combinations. Calcium channel blockers (CCBs) are also effective antihypertensive agents, and evidence suggests that a CCB/ACEI combination is well tolerated and also decreases the risk of cardiovascular and renal disease. Some evidence suggests that this combination may improve endothelial function more than either agent alone, and its use could potentially lead to better cardiovascular outcomes than a diuretic/ACEI or diuretic/ARB combination. The ongoing Avoiding Cardiovascular Events Through Combination Therapy in Patients Living With Systolic Hypertension (ACCOMPLISH) trial compares these 2 effective combinations (ie, an ACEI/diuretic and ACEI/CCB) as initial treatment for reducing cardiovascular morbidity and mortality in older high-risk hypertensive patients. The results of this trial, when reported, should help to clarify the relative benefits of these different therapies. PMID- 17917507 TI - Hypertension and coronary artery disease: a summary of the American Heart Association scientific statement. AB - The American Heart Association scientific statement on the treatment of hypertension in the prevention and management of ischemic heart disease was published recently. The main recommendations were as follows: (1) For most adults with hypertension, the blood pressure (BP) goal is <140/90 mm Hg but should be <130/80 [corrected] mm Hg in patients with diabetes mellitus, chronic kidney disease, known coronary artery disease (CAD), CAD equivalents (carotid artery disease, abdominal aortic aneurism, and peripheral vascular disease), or 10-year Framingham risk score of >/=10%. For those with left ventricular dysfunction, the recommended BP target is <120/80 mm Hg. (2) For primary CAD prevention, any effective antihypertensive drug or combination is indicated, but preference is given to angiotensin-converting enzyme (ACE) inhibitors, angiotensin receptor blockers (ARBs), calcium channel blockers (CCBs), and thiazide diuretics. (3) For the management of hypertension in patients with established CAD (stable or unstable angina, non-ST-segment elevation myocardial infarction, ST-segment elevation myocardial infarction), beta-blockers and ACE inhibitors (or ARBs) are the basis of treatment. If further BP lowering is needed, a thiazide diuretic and/or a dihydropyridine CCB (not verapamil or diltiazem) can be added. If a beta blocker is contraindicated or not tolerated, diltiazem or verapamil can be substituted. (4) If there is left ventricular dysfunction, recommended therapy consists of an ACE inhibitor or ARB, a beta-blocker, and either a thiazide or loop diuretic. In patients with more severe heart failure, an aldosterone antagonist and hydralazine/isosorbide dinitrate (in black patients) should be considered. PMID- 17917508 TI - Enhancing dosage flexibility in combination therapy. PMID- 17917509 TI - Yoga and hypertension. PMID- 17917510 TI - Maximizing diuretic therapy in resistant hypertension. PMID- 17917511 TI - The natural history of hypertension: prehypertension or masked hypertension? PMID- 17917512 TI - Fine-tuning blood pressure control to minimize cardiovascular risk. PMID- 17917513 TI - Casual chocolate consumption and inhibition of platelet function. AB - Observational studies have associated reduced cardiovascular mortality with chocolate consumption. Feeding studies of high-dose, flavanol-rich chocolate show antiplatelet effects, but the effect of casual chocolate consumption on platelet function is unknown. Healthy adults (N=1535) were proscribed from consuming foods affecting platelet function, including chocolate, for 48 hours and completed a 24 hour dietary recall before ex vivo platelet testing with the Platelet Function Analyzer (PFA)-100 (Dade Behring, Inc, Deerfield, IL) test and in vivo testing with urinary 11-dehydro thromboxane B2 (Tx-M) measurements. Some participants (n=141) reported ignoring the prohibition of consuming chocolate before platelet testing. Despite having similar baseline characteristics, chocolate consumers had longer PFA closure times (130 vs 123 seconds, P=.005) and decreased Tx-M levels (175 vs 290 ng/mol creatinine, P=.03). Chocolate remained a significant independent predictor of both ex vivo and in vivo platelet function testing after adjusting for confounders. The authors concluded that even consuming modest amounts of commercial chocolate has important antiplatelet effects. PMID- 17917514 TI - Distribution and predictors of carotid intima-media thickness in young adults. AB - The authors' aim was to determine reference values and predictors for carotid intima-media thickness (CIMT) in the Bogalusa Heart Study. Carotid ultrasound images from 1203 young adults (mean age 36 years; 30% black, 43% male) were reviewed. Age-, sex-, and race-specific CIMT percentiles were estimated using multivariable regression. Nomograms of CIMT quartiles for persons aged 25 to 40 years are provided in 5-year increments. CIMT was thickest in the carotid bulb and increased linearly with age, most rapidly in the bulb. With age, composite CIMT increased most slowly in white women and more rapidly in white men and black women. Systolic blood pressure (P<.001) was the strongest predictor of composite CIMT. Male sex, total to high-density lipoprotein cholesterol ratio, and age independently predicted CIMT in all segments. This report may be used to help plan epidemiologic investigations and clinical trials investigating atherosclerosis and its changes with interventions. PMID- 17917516 TI - Discordant hemodynamic and fibrinolytic adaptations following a 6-week cardiac rehabilitation program. AB - The present study evaluated changes in hemodynamics and fibrinolysis during 6 weeks of participation in an exercise-based cardiac rehabilitation program. Fourteen patients trained for 3 days per week for 6 weeks using American College of Sports Medicine guidelines for intensity and duration. Blood samples were taken at baseline and after 3 and 6 weeks of participation and analyzed for tissue plasminogen activator (t-PA) activity and antigen, plasminogen activator inhibitor-1 (PAI-1) activity and antigen, and relative quantification of t-PA and PAI-1 RNA. Data were analyzed using repeated measures analysis of variance. Training elicited significant decreases in submaximal exercise heart rate and systolic blood pressure and resting systolic blood pressure (p<.05). There were no significant changes in plasma concentrations of t-PA or PAI-1, and no change was observed in t-PA or PAI-1 gene expression. The present findings suggest that favorable hemodynamic adaptations may occur after only 6 weeks of exercise-based cardiac rehabilitation, but longer training periods may be needed to elicit positive hemostatic adaptations. PMID- 17917515 TI - Cardiovascular disease knowledge, medication adherence, and barriers to preventive action in a minority population. AB - Lack of knowledge and awareness of cardiovascular disease may contribute to disproportionately higher risk in minorities. The authors studied minorities in Harlem, New York (N=214), to evaluate knowledge and preventive behaviors. More than half of the participants did not know optimal blood pressure (BP) (52%) and cholesterol (60%) goals. Lack of health insurance (odds ratio, 2.1; 95% confidence interval, 1.0-4.5) and less than a high school education (odds ratio, 2.0;95% confidence interval, 1.02-3.87) were associated with not knowing optimal BP. Among those with BP >/=140/90 mm Hg, 34% were unaware that they had high BP, and age younger than 55 years was predictive of lack of awareness that they had high BP (odds ratio, 8.5; 95% confidence interval, 2.6-28.1). Predictors of medication nonadherence included age younger than 45 years vs age 45 years or older (P=.004) and no health insurance vs health insurance (P=.01). Younger, less educated, uninsured patients should be targeted for educational interventions regarding cardiovascular disease prevention goals, personal risk, and the importance of medication adherence. PMID- 17917517 TI - The role of quinapril in the presence of a weight loss regimen: endothelial function and markers of obesity in patients with the metabolic syndrome. AB - Forty-four patients with the metabolic syndrome were placed on a reduced-calorie and reduced-fat regimen to lose weight throughout a 56-week period. The patients were treated in a crossover fashion with placebo and the angiotensin-converting enzyme inhibitor quinapril for 24 weeks each. The study measured endothelial dependent flow-mediated dilation plus serum obesity markers of adiponectin and leptin. Metabolic parameters improved after 56 weeks. Serum adiponectin level increased by 18% (P<.05 vs baseline) and serum leptin level decreased by 16% with placebo (P<.05 vs baseline). These findings were potentiated further in the quinapril group. In comparison with baseline, flow-mediated dilation was increased by 13% in the placebo group (P=.055 vs baseline) and by 43% in the quinapril group (P<.001 vs baseline and placebo). These findings suggest that weight loss therapy improves endothelial function and markers of obesity. These results are potentiated with quinapril and are independent of changes in metabolic parameters. PMID- 17917518 TI - Clinical assessment of early morning blood pressure in patients with hypertension. AB - In most individuals with hypertension, blood pressure (BP) shows a moderate to marked increase around the time of awakening, which has been linked to increases in cardiovascular complications occurring at this time of day. Many antihypertensive agents do not adequately control early morning BP, particularly when administered once daily in the morning. Points to consider in selecting an effective antihypertensive drug include pharmacokinetics and formulation of the agent and timing of administration. Agents with long pharmacologic half-lives, such as the angiotensin II receptor blocker telmisartan, the calcium antagonist amlodipine, and the beta-blocker bisoprolol, are examples of antihypertensive drugs with demonstrated efficacy in controlling early morning BP. Bedtime administration of chronotherapeutic preparations is also effective for controlling early morning BP. Given the association between early morning BP and cardiovascular risk, future clinical trials should focus on the efficacy of antihypertensive drugs during this important period of risk. PMID- 17917520 TI - Where are we going? Beyond JNC 7. AB - On August 15, 2007, a panel discussion was held to discuss hypertension. The panel was moderated by Thomas D. Giles, MD, Tulane University School of Medicine, New Orleans, LA. Discussants included Suzanne Oparil, MD, University of Alabama at Birmingham, Birmingham, AL, and Thomas G. Pickering, MD, DPhil, Columbia Presbyterian Medical Center, New York, NY. PMID- 17917521 TI - The role of advanced lipid testing in clinical practice. AB - The National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) guidelines recommend assessing an individual's cardiovascular (CV) risk from the Framingham risk score; however, the Framingham risk score may underestimate coronary heart disease (CHD) risk. Current guidelines have identified some emerging lipid risk factors that can be measured by several commercially available advanced cholesterol tests. These emerging lipid risk factors are meant to supplement the Framingham risk score to help the clinician to better assess CV risk. Although advanced lipid testing cannot be recommended for routine screening, it may be of value in individuals with a family history of premature CHD, postmenopausal women, and individuals at intermediate risk for CHD, especially if they are near the boundary of being at high risk. This review examines the role of advanced lipid testing in clinical practice. PMID- 17917519 TI - Reducing the risk of stroke through appropriate targets and treatments. AB - On June 29, 2007, Michael H. Davidson, MD, of the University of Chicago, Chicago, IL, moderated a panel of participants, including Jan Basile, MD, Primary Care Service Line, Ralph H. Johnson VA Medical Center, and the Division of General Internal Medicine/Geriatrics, Medical University of South Carolina, Charleston, SC; Alan J. Garber, MD, PhD, Baylor College of Medicine, Houston, TX; and Robert A. Phillips, MD, PhD, University of Massachusetts Memorial Medical Center, Worcester, MA. With the publication of this first Expert Panel Discussion in Preventive Cardiology, we inaugurate a new educational feature. Periodically, we will gather a group of experts to exchange viewpoints on current issues relevant to the prevention of cardiovascular disease. It is our hope that the resulting discussion will provide the practicing clinician with data from recent trials, expert opinions on symptoms and diagnoses, and a variety of potential therapeutic approaches for the management of risk factors. PMID- 17917522 TI - The rosiglitazone controversy: meta-analysis and the RECORD study. PMID- 17917523 TI - American society for preventive cardiology. PMID- 17917525 TI - PRO: Should aspirin be used in all women older than 65 years to prevent stroke? AB - A number of studies have reported that aspirin is beneficial in the prevention of cardiovascular disease. A meta-analysis of data from 3 studies of the cardioprotective effect of aspirin in women has reported that use of aspirin reduces the risk of coronary events mainly by reducing the risk of ischemic stroke. Results of the Women's Health Study showed that although there is a risk of bleeding events with aspirin use, overall this risk is outweighed by the number of strokes prevented. PMID- 17917526 TI - CON: Should aspirin be used in all women older than 65 years to prevent stroke? AB - A goal of the Women's Health Study was to evaluate the balance of benefits and risks of low-dose aspirin in the primary prevention of stroke in healthy women. A total of 39,876 female health care professionals aged 45 years and older were randomly assigned to either low-dose aspirin (100 mg every other day) or placebo and followed for an average of 10 years. In the overall study population, aspirin significantly lowered the risk of total and ischemic stroke without affecting the risk of myocardial infarction or death from cardiovascular causes, leading to a nonsignificant finding with respect to the primary end point of total cardiovascular disease. In women aged 65 years or older, aspirin significantly reduced the risk of ischemic stroke, as well as myocardial infarction and total cardiovascular disease. While the findings suggest that aspirin should be considered for primary prevention of stroke in women aged 65 years or older, the balance of benefits and risks would not support recommending aspirin for all women in this age group. The discussion on whether to use aspirin should be had on an individual basis, assessing the net risks and benefits. PMID- 17917527 TI - Guidelines for prevention of cardiovascular disease in women: a summary of recommendations. AB - Cardiovascular disease (CVD) is a major cause of mortality in women. The first evidence-based guidelines for the prevention of CVD in women were developed in 2004 and updated this year. The 2007 guidelines place more emphasis on women's overall lifetime coronary heart disease risk, as opposed to short-term absolute risk, than did the 2004 recommendations. Depending on the criteria met, women are classified as being at high risk, at risk, or at optimal risk. The authors provide an overview of the recommendations in the 3 different categories: lifestyle interventions, major risk factor interventions, and preventive drug interventions. PMID- 17917528 TI - Does an aspirin a day really keep a stroke away? PMID- 17917530 TI - Autoantibodies in idiopathic inflammatory myopathy: an update on clinical and pathophysiological significance. AB - PURPOSE OF REVIEW: Idiopathic inflammatory myopathy is characterized by the production of autoantibodies to various cellular constituents. These autoantibodies closely correlate with certain clinical conditions and prognosis of disease. This review examines recent progress in myositis-specific autoantibodies, particularly in their clinical significance and pathophysiological roles. RECENT FINDINGS: During the 1-year review period, novel myositis-specific autoantibodies were identified in clinically amyopathic dermatomyositis (anti-CADM-140 antibody) and malignancy-associated myositis (anti p155 and anti-p155/p140 antibodies). These new autoantibodies are extremely important because it is thought that myositis-specific autoantibodies are negative in these subgroups, and may enable a new classification of idiopathic inflammatory myopathy. New clinical aspects of other myositis-specific autoantibodies (anti-aminoacyl-tRNA synthetases, anti-signal recognition particles and anti-Mi-2) are also described. The possibility was raised that the high expression of myositis-specific autoantigens in regenerating muscle cells and certain cancers may be involved in initiating and perpetuating the autoimmune response in myositis. SUMMARY: Myositis-specific autoantibodies are useful markers for clinical diagnosis, classification and predicting prognosis of idiopathic inflammatory myopathy. To understand the etiopathogenic mechanisms of the disease it is particularly important to elucidate the nature of target autoantigens recognized by these myositis-specific autoantibodies. PMID- 17917531 TI - Skeletal muscle imaging and inflammatory myopathies. AB - PURPOSE OF REVIEW: A variety of modalities exist for the imaging of skeletal muscle including radiography, ultrasound, computed tomography, and MRI. This article highlights the utility of these modalities in evaluating skeletal muscle diseases. Newer techniques such as T2 mapping, blood oxygenation level dependent imaging, diffusion tensor imaging, and magnetic resonance spectroscopy are also explored. RECENT FINDINGS: Cross-sectional imaging including computed tomography and magnetic resonance is the current standard in skeletal muscle imaging. The advantages of these modalities include the ability to image in different planes and to evaluate the distribution of disease and disease burden. Newer magnetic resonance-based techniques also provide functional information in addition to anatomic information. Radiography and ultrasound have a more limited role and are mainly used to detect calcifications and evaluate the texture of skeletal muscle. SUMMARY: Magnetic resonance is a useful modality for evaluating skeletal muscle and allows for the assessment of disease burden. It can elucidate potential targets for biopsy. Newer magnetic resonance techniques can acquire functional information in addition to anatomic information and hold tremendous potential for detecting, characterizing, and monitoring treatment for inflammatory myopathies. PMID- 17917532 TI - A gene expression approach to study perturbed pathways in myositis. AB - PURPOSE OF REVIEW: To review new insights into the disease mechanisms of dermatomyositis, inclusion body myositis, and polymyositis gained from large scale microarray gene expression studies of patient tissue samples. RECENT FINDINGS: The detection of unexpected gene transcripts using microarrays in inflammatory myopathy tissue has led to the discovery of new types and roles of immune system cells present in muscle in these diseases. Plasmacytoid dendritic cells, the immune system's professional producer of the type 1 interferons alpha and beta, are prominent in dermatomyositis muscle. Plasma cells and myeloid dendritic cells are abundant in polymyositis and inclusion body myositis muscle. Type 1 interferon induction is the single most upregulated pathological pathway genome-wide in dermatomyositis muscle and blood. In individual patients with dermatomyositis and some with polymyositis, a blood type 1 interferon-signature correlates with disease activity. SUMMARY: The identification of new cells and pathways in inflammatory myopathies has led to deeper mechanistic understanding of these diseases and potential therapeutic approaches. Through insights gained in gene expression studies, a strong scientific rationale has developed for blockade of the type 1 interferon pathway for treatment of dermatomyositis. PMID- 17917533 TI - Skeletal muscle remodeling. AB - PURPOSE OF REVIEW: In skeletal muscle, environmental demands activate signal transduction pathways that ultimately promote adaptive changes in myofiber cytoarchitecture and protein composition. Recent advances in determining the factors involved in these signal transduction pathways provide insight into possible therapeutic methods to remodel skeletal muscle. RECENT FINDINGS: Advances in genetic engineering have allowed the introduction or depletion of factors within the myofiber, facilitating the evaluation of signaling factors during muscle remodeling. Using transgenic mouse models, activation of specific signaling pathways promoted type I oxidative myofibers, increased the fatigue resistance of muscle, increased skeletal muscle mass and ameliorated muscle injury in myopathic mouse models. Moreover, new technologies are being used to generate global gene and protein expression profiles to identify new factors involved in skeletal muscle remodeling. Finally, small RNAs, microRNAs, are emerging as powerful regulators of gene expression in most tissues, including skeletal muscle. Recent findings predict that targeted delivery of miRNAs will specifically manipulate genes and if used therapeutically will revolutionize clinical medicine. SUMMARY: Developing drugs to target signaling pathways associated with remodeling myofibers provides a possible therapeutic approach to combat skeletal muscle disease. In addition, genome-wide technologies can identify new biomarkers capable of diagnosing myopathies and determine a patient's response to therapy. Furthermore, therapeutic strategies are being designed to target microRNAs in anticipation of blocking gene repression correlated with muscle pathology. PMID- 17917534 TI - Inclusion-body myositis, a multifactorial muscle disease associated with aging: current concepts of pathogenesis. AB - PURPOSE OF REVIEW: Sporadic inclusion-body myositis, the most common muscle disease of older persons, has no known cause or persistently beneficial treatment. The unfolding pathogenesis could lead to new treatment strategies and it is now of growing interest among clinicians and basic scientists. About 100 papers related to the subject were published in 2006 and the first part of 2007 (we cite only articles most relevant to this review). RECENT FINDINGS: This review focuses on the current concepts of the pathogenesis of sporadic inclusion body myositis. Both degeneration and mononuclear-cell inflammation are components of the pathology, but how each relates to the pathogenesis remains unclear. We suggest that an intramuscle fiber degenerative component is primary, leading to muscle-fiber destruction, while the lymphocytic inflammatory component may only slightly contribute to sporadic inclusion-body myositis muscle-fiber damage. Intracellular accumulation of amyloid-beta precursor protein, amyloid-beta, and amyloid-beta oligomers in an aging muscle-fiber cellular milieu, and other abnormalities, appear to be key pathogenic factors. We summarize intracellular molecular events and their consequences, and correlate findings in sporadic inclusion-body myositis muscle biopsies with inclusion-body myositis experimental models in tissue culture and in transgenic mice. SUMMARY: Treatment of sporadic inclusion-body myositis remains a challenge. Antiinflammatory approaches used so far are without major or enduring benefit. Possible new treatment avenues are suggested. PMID- 17917536 TI - Understanding systemic sclerosis through gene expression profiling. AB - PURPOSE OF REVIEW: Genomic analysis has rapidly become commonplace in the study and treatment of complex disease. Several recent studies of gene expression profiling in systemic sclerosis have demonstrated its value in diagnosis and illustrate the potential for this technique in prognostication, as well as the elucidation of the underlying pathogenesis. RECENT FINDINGS: Skin biopsies from patients with systemic sclerosis show robust changes in gene profile that precede clinically detectable involvement. Current results suggest that clinically indistinguishable subgroups may be identified with different pathogenesis and outcome. Expression profiling studies of animal models of systemic sclerosis and explanted fibroblasts have helped to reveal the utility and deficiencies of these surrogates in the study of systemic sclerosis. SUMMARY: Gene profiling is likely to provide valuable prognostic information in systemic sclerosis patients. Recent advances in sample collection and standardization of analysis mean that longitudinal collection of samples for gene profiling, even in small numbers of patients from different clinical centers, will contribute enormously to our understanding of the disease. PMID- 17917537 TI - Molecular pathways as novel therapeutic targets in systemic sclerosis. AB - PURPOSE OF REVIEW: Systemic sclerosis is a complex disease characterized by immune/inflammatory, vascular and fibrotic processes. To date, no treatment has proven effective in modifying the course of the disease. Recent studies have begun to yield insights into the nature and interrelationship among these processes, and their cellular and molecular components. RECENT FINDINGS: Novel intracellular molecular pathways have been characterized that positively or negatively regulate fibroblast responses contributing to the process of fibrosis. These include signaling mediators that specify and amplify transforming growth factor-beta responses, or inhibit collagen stimulation and block these responses in vitro and in animal models. Various gain of function or loss of function abnormalities in these mediators have been identified in systemic sclerosis, and may account for the characteristic activated phenotype of systemic sclerosis fibroblasts. SUMMARY: The identification of novel signaling pathways and mediators that are altered in systemic sclerosis and contribute to tissue damage allows their selective targeting. This in turn opens the door for novel therapeutic strategies utilizing novel compounds, or innovative ways of using already-approved drugs. In light of the complex pathogenesis of systemic sclerosis, however, only carefully designed clinical trials with appropriate biomarkers and outcome measures will be able to clarify the clinical utility of these innovative approaches. PMID- 17917538 TI - Platelet contributions to the pathogenesis of systemic sclerosis. AB - PURPOSE OF REVIEW: The purpose of this review is to focus attention on platelet contributions, in general, to systemic sclerosis. There have also been recent advances in characterization of the phenotype of platelets in systemic sclerosis which will be reviewed. RECENT FINDINGS: An extensive literature provides strong support for varying degrees of platelet activation and aggregation in different forms and stages of systemic sclerosis. A recent finding is that systemic sclerosis platelets overexpress a specific nonintegrin 65 kDa receptor for type I collagen as well as expressing enhanced phosphilidylinositol-3 kinase as an activation signature. Overexpression of a type I collagen receptor would make systemic sclerosis platelets more susceptible to binding to exposed type I collagen in the subendothelial lining of damaged blood vessels, facilitating the cycle of platelet aggregation and release of preformed bioactive molecules that include a host of inflammatory and fibrogenic, chemokines, cytokines and growth factors. This activation phenotype of systemic sclerosis platelets may be secondary to autoimmunity and driven by cytokines from autoreactive T cells. SUMMARY: The contributions of platelets to the pathogenesis of systemic sclerosis is likely substantial and may not be adequately represented in gene profiling of systemic sclerosis tissue due to the small amounts of RNA contained in platelets. PMID- 17917539 TI - Update on autoantibodies in systemic sclerosis. AB - PURPOSE OF REVIEW: This is a review of autoantibodies described in systemic sclerosis with an emphasis on recently published studies. In the past, most, if not all of the discussion on this topic focused on antinuclear antibodies, but it is now appreciated that autoantibodies to cytoplasmic, cell surface, intercellular and plasma components are also important in the context of systemic sclerosis. RECENT FINDINGS: A number of recent studies have highlighted the disease associations of autoantibodies and the potential pathogenic role of the more traditional autoantibodies, such as antitopoisomerase I and anticentromere antibodies. The recent identification of autoantibodies directed to the platelet derived growth factor receptor is of particular interest because of its possible association with the pathogenesis of systemic sclerosis. SUMMARY: Autoantibodies in systemic sclerosis are associated with demographic, diagnostic, pathological, and prognostic features of the disease. Emerging research on the pathogenic roles of newer autoantibodies provides valuable insights into disease pathogenesis and potential therapeutic targets. PMID- 17917540 TI - Childhood systemic sclerosis. AB - PURPOSE OF REVIEW: Juvenile systemic sclerosis has a variety of clinical manifestations, sometimes different from the adult form. Early recognition, proper classification and treatment may improve the long-term outcome. RECENT FINDINGS: A large multicenter study coordinated by the Pediatric Rheumatology European Society has yielded important information on the epidemiology and clinical manifestations of systemic sclerosis in childhood. An ad-hoc Committee on Classification Criteria for Juvenile Systemic Sclerosis developed the new classification criteria to help improve patient care by enabling earlier, more definite diagnoses and standardizing the conduct of clinical, epidemiologic, and outcome research for this rare disease. The overall outcome of children with systemic sclerosis is better than in adults but, in those cases with a fatal course, disease progression is rapid and an early involvement of internal organs is associated with poor outcome. SUMMARY: Studies over the past few years have highlighted the peculiar clinical features and the better outcome of juvenile systemic sclerosis compared with the adult form and propose new pediatric classification criteria. Efforts have recently been made to address the definition of evidence-based recommendations for the treatment of adult and pediatric onset systemic sclerosis. PMID- 17917541 TI - Screening and therapy of pulmonary hypertension in systemic sclerosis. AB - PURPOSE OF REVIEW: Systemic sclerosis is a major risk factor for the development of pulmonary arterial hypertension. Often this disease is far advanced by the time it is detected in the individual patient, severely impacting quality of life and survival. RECENT FINDINGS: Therapeutic options available for patients with scleroderma-associated pulmonary arterial hypertension have increased significantly over the last few years, fueling our desire to improve early detection of disease. SUMMARY: During the course of this manuscript I will review the literature examining approaches towards screening for scleroderma-associated pulmonary arterial hypertension. I will then discuss the most recent studies examining therapeutic options for patients with this disease. PMID- 17917542 TI - Adult stem cell treatment of scleroderma. AB - PURPOSE OF REVIEW: Provides an update of hematopoietic stem cell transplantation for systemic sclerosis from phase I/II studies and prospective randomized phase III trials, and introduces the concept of mesenchymal stem cells as potential therapy for autoimmune disease. RECENT FINDINGS: Around 170 transplanted systemic sclerosis patients are registered in Europe. Most received autologous, peripheral blood derived hematopoietic stem cell transplantation. Treatment-related mortality has fallen to 2.5% in the controlled trials compared with 12.5% in the first report in 2002. Over one-third of patients have experienced sustained remission. Two prospective randomized phase III studies are active: the Autologous Stem cell Transplantation International Scleroderma (ASTIS) trial in Europe and the Scleroderma Cyclophosphamide Or Transplant (SCOT) trial in the USA. Both have similar selection criteria, endpoint and control arms, but the SCOT trial uses radiation and less cyclophosphamide. So far, no unexpected toxicity has occurred. Reports produced in the past 12 months show reduction of skin collagen and reversal of microvascular remodelling, years after transplant. Bone marrow-derived mesenchymal stem cells from systemic sclerosis patients show in-vitro immunomodulatory properties equal to healthy controls. SUMMARY: Hematopoietic stem cell transplantation is currently being tested in prospective randomized controlled trials and appears to 'reset' autoimmunity in systemic sclerosis. Mesenchymal stem cells may have an immunomodulatory role in autoimmune disease. PMID- 17917543 TI - Current drug therapy for scleroderma and secondary Raynaud's phenomenon: evidence based review. AB - (1) Scleroderma and secondary Raynaud's phenomenon are frequently associated with increased morbidity for which no specific standardised treatment guidelines exist. (2) Current therapies for scleroderma target the immune system, with the goal of reducing inflammation and secondary tissue injury and fibrosis. Therapy targeting underlying vascular disease is designed to improve the symptoms of Raynaud's phenomenon and to reduce ischemic injury to involved organs. (3) Few controlled trials of therapy used for scleroderma are completed, and current treatments are largely based on organ-specific therapy and uncontrolled case series suggesting disease modification. (4) Recent randomised, controlled trials in scleroderma demonstrate promising results in the treatment of interstitial lung disease with cyclophosphamide, and vascular disease of the lungs and digits with endothelin receptor antagonists, the phosphodiesterase inhibitor sildenafil and prostacyclins, while trials with methotrexate show only modest benefit in controlling scleroderma-associated skin disease. (5) Prostacyclins are a therapeutic option in patients with secondary Raynaud's phenomenon. Modest benefits have also been shown with alpha1-antagonists and calcium channel blockers, while the effect of ACE inhibitors has been variable. Some data suggest some benefits to the use of the phosphodiesterase inhibitor sildenafil, the serotonin uptake inhibitor fluoxetine and the angiotensin receptor inhibitor losartan. PMID- 17917544 TI - Risks of immunosuppressive therapies including biologic agents in patients with rheumatic diseases and co-existing chronic viral infections. AB - PURPOSE OF REVIEW: A number of chronic viral infections could be reactivated by immunosuppressive agents used in rheumatic diseases. In this review, we will focus on the complex effect of immunosuppressive agents, including biologic agents, on the natural course of chronic viral infections as well as an approach to the prevention and management of therapy-induced viral reactivation. RECENT FINDINGS: Chronic viral infections that are affected by immunosuppression in the setting of an underlying rheumatic disease include those due to hepatitis B virus, hepatitis C virus, or human immunodeficiency virus, and latent infections from Epstein-Barr virus, JC virus, or varicella zoster virus. The most recent data of the effects of immunosuppressive agents are reviewed, with special emphasis on the effects of biologic therapies (anti-tumor necrosis factor, anti-B cell), on these viral agents. SUMMARY: Clinicians should be aware of the risk for viral reactivation of an underlying chronic viral infection during immunosuppressive therapy. Despite the existence of such risk, the presence of chronic viral infection is not a contraindication to immunosuppressive therapy, given that appropriate pretherapy screening and close monitoring is applied. PMID- 17917545 TI - Targets of anticytokine therapy and the risk of infections in humans and mice. AB - PURPOSE OF REVIEW: To examine the type and risk of infections in humans and mice deficient in proinflammatory cytokines. Naturally occurring or manipulated genetic defects of tumor necrosis factor, interleukins-1, -6, -12, and -15, and interferon-gamma are examined for their increased susceptibility to, or protection from, infection. RECENT FINDINGS: Interleukin-12p40 and interferon gamma-blockers may lead to increased incidence of infections with intracellular bacteria, parasites, and fungi. In addition, we may see viral infections with interferon-gamma-blockers. Increased risk of infections is unlikely with either interleukin-1- or interleukin-15-blockers. Interleukin-6-blockers may lead to increased risk of infection with extracellular bacteria, viruses, parasites and fungi. SUMMARY: In tumor necrosis factor knockout mice, increased susceptibility to pathogens are reported that are normally controlled by granuloma formation. In patients treated with tumor necrosis factor-blockers, a two-fold increase of granulomatous infections, predominantly reactivation of latent tuberculosis, is found. The infections detected in tumor necrosis factor knockout mice were accurate for predicting the infections observed when using tumor necrosis factor blockers. If a similar correlation exists for other cytokines, the use of interferon-gamma and interleukin-12p40 blockers, and possibly interleukin-6 blockers, will lead to an increased risk for severe infections. Care should be taken when new cytokine blockers/antagonists are introduced. PMID- 17917546 TI - Viral infection can induce the production of autoantibodies. AB - PURPOSE OF REVIEW: To review the current literature and summarize the main principles found between viral infections and the subsequent production of autoantibodies. RECENT FINDINGS: We concentrate on recent findings involving three viral agents, one of which is Epstein-Barr virus, which has been associated with many autoimmune diseases and is classically considered to induce systemic lupus erythematosus. As we will discuss, this occurs through molecular mimicry between Epstein-Barr virus nuclear antigen 1 and lupus-specific antigens such as Ro, La or dsDNA, through induction of Toll-like receptor hypersensitivity by Epstein-Barr virus latent membrane protein 2A or by creating immortal B and T cells by loss of apoptosis. Hepatitis B virus was found to share amino acid sequences with different autoantigens. Tissue damage and the release of intracellular components is just another example of the autoantibody production caused by this virus. Cytomegalovirus has often been controversially associated with several autoimmune diseases and, although is the least understood viral infection of the three, appears to be somewhat suspicious. SUMMARY: Understanding the infectious origin of autoimmune diseases is important as we aim to identify high-risk patients and disrupt this process with vaccines or other medications, ultimately delaying or even preventing the evolution of autoimmune diseases. PMID- 17917548 TI - Bibliography. Current world literature. Myositis and myopathies. PMID- 17917547 TI - Severe congenital neutropenia: new genes explain an old disease. AB - PURPOSE OF REVIEW: This review summarizes the recent advances in the diagnosis and molecular characterization of isolated and syndromal forms of severe congenital neutropenia. RECENT FINDINGS: It has become evident that severe congenital neutropenia comprises several genetically distinct entities. In 1999, mutations were identified in the neutrophil elastase gene ELA2. ELA2 mutations have been found in cyclic, sporadic and autosomal dominant neutropenia. Recently, homozygous mutations in the antiapoptotic gene HAX1 were found in patients with autosomal recessive severe congenital neutropenia. Ongoing linkage studies suggest that more and, as yet unidentified, genes may be involved in the pathophysiology of severe congenital neutropenia. In other patients, congenital neutropenia is not an isolated finding but is associated with other abnormalities, in particular, lymphoid immunodeficiency and pigmentation defects such as Chediak-Higashi syndrome, Griscelli syndrome type 2, Hermansky-Pudlak syndrome type 2, or deficiency of the endosomal adaptor p14. The molecular identification of these disorders originating from mutations in lysosome (related) proteins has advanced our knowledge of intracellular protein trafficking. SUMMARY: Recent insights into the molecular etiology of severe congenital neutropenia provide the opportunity for a definitive genetic classification system. Based on this knowledge, disease-related risks may be recognized and optimized therapeutic options may become available. PMID- 17917549 TI - Sustained response versus relapse: the pharmacotherapeutic goal for obsessive compulsive disorder. AB - Convincing evidence from placebo-referenced randomized controlled trials supports efficacy for clomipramine and selective serotonin reuptake inhibitors for acute treatment of obsessive-compulsive disorder. It remains less conclusively understood whether these agents maintain efficacy over the longer term. This paper systematically reviews long-term medication studies in obsessive-compulsive disorder. Studies of clomipramine, fluoxetine and sertraline investigated 'responders' from acute treatment trials and extended treatment up to 12 months versus placebo. Responses to medication were sustained. A 24-week placebo controlled trial of escitalopram (10 mg or 20 mg/day) and paroxetine (40 mg/day) demonstrated ongoing efficacy for all three treatments. Studies that randomized treated cases to placebo demonstrated reemergence of symptoms in the placebo treated cohort. Six relapse prevention trials were found by systematic search. Some, but not all, revealed significant advantages for remaining on medication. Paroxetine (20-60 mg/day) and escitalopram (10 or 20 mg/day) were each found to outperform placebo in preventing relapse during 24 weeks of double-blind, randomized follow-up. Meta-analysis, using Review Manager software (4.2.8), detected overall superiority of selective serotonin reuptake inhibitors to placebo in preventing relapse among adult treatment-responders. Worsening by five Yale-Brown Obsessive Compulsive Scale points emerged from the review as a suggested threshold for relapse. Viewed collectively, these results suggest that selective serotonin reuptake inhibitors are effective long-term treatments and relapse prevention represents the treatment target for obsessive-compulsive disorder. PMID- 17917550 TI - Which antidepressants have demonstrated superior efficacy? A review of the evidence. AB - A review of published evidence of superior efficacy of a particular antidepressant in major depressive disorder may assist clinicians in making considered treatment choices. To identify such candidates, an international group of experts met to assess published evidence (identified through searches in Medline and Embase databases and discussions with experts in the field) from randomized, controlled trials and meta-analyses comparing two antidepressants under conditions of fair comparison. Criteria were defined to judge the strength of evidence. Two pivotal studies in moderate-to-severe major depressive disorder that demonstrate superiority on the primary efficacy measure, or alternatively one pivotal study supported by consistent results from meta-analyses, was considered to constitute evidence for definite superiority. Three antidepressants met these criteria: clomipramine, venlafaxine, and escitalopram. Three antidepressants were found to have probable superiority: milnacipran, duloxetine, and mirtazapine. Only escitalopram was found to have definite superiority in the treatment of severe depression; probable superiority was identified for venlafaxine and possible superiority for milnacipran and clomipramine. This review of published data found evidence that only a very few antidepressants are shown to be more effective than others. PMID- 17917551 TI - Valproate-induced hyperammonaemic encephalopathy: review of 14 cases in the psychiatric setting. AB - OBJECTIVES: To review signs and symptoms of valproate-induced hyperammonaemic encephalopathy without hepatotoxicity in the psychiatric setting, explore its mechanisms, and give recommendations for prevention and treatment. METHODS: Medline search with keywords valproate, ammonia, hyperammonaemia, encephalopathy, and then cross-references to articles obtained through this search. Only cases with indication of valproate for psychiatric condition were included. RESULTS: Fourteen cases published in the psychiatric setting are reviewed. Valproate induced hyperammonaemic encephalopathy is a rare adverse event, occurring almost equally in men and women, with a large age range, and reported in two patients with mental retardation. Symptoms appeared either a few days after initiation of valproate therapy, or after several months or years. The main symptoms were fluctuations in consciousness and disorientation. Clinical severity was not related to blood ammonia levels. All patients recovered after valproate-induced hyperammonaemic encephalopathy diagnosis and treatment, usually involving discontinuation of valproate. CONCLUSIONS: Valproate-induced hyperammonaemic encephalopathy is rare and usually reversible in patients without urea cycle disorders when valproate is discontinued. Therapy with carnitine is recommended. Special caution should be used in patients with mental retardation. Psychiatrists should suspect valproate-induced hyperammonaemic encephalopathy when consciousness deteriorates. PMID- 17917552 TI - A randomized, double-blind, placebo-controlled trial of desvenlafaxine succinate in the treatment of major depressive disorder. AB - The antidepressant efficacy and safety of desvenlafaxine succinate (desvenlafaxine) were evaluated in a phase III, double-blind, placebo-controlled study. Outpatients with a primary diagnosis of major depressive disorder were treated with fixed once-daily doses of desvenlafaxine 200 or 400 mg for 8 weeks. The primary efficacy measure was change from baseline on the 17-item Hamilton Rating Scale for Depression. At the final on-therapy evaluation, adjusted mean change from baseline in 17-item Hamilton Rating Scale for Depression total score was greater for desvenlafaxine 200 and 400 mg/day vs. placebo. Both desvenlafaxine doses showed greater efficacy than placebo on the secondary efficacy measures, including the Clinical Global Impressions-Improvement scale scores, Montgomery-Asberg Depression Rating Scale scores, CGI-Severity, and 17 item Hamilton Rating Scale for Depression response rate. Desvenlafaxine 200 mg/day was also significantly better than placebo on remission, Visual Analog Scale-Pain Intensity overall scores, and some Visual Analog Scale-Pain Intensity subscale scores. Desvenlafaxine 400 mg/day was significantly better than placebo on selected Visual Analog Scale-Pain Intensity subscale scores. Most adverse events were mild or moderate in severity, and safety assessments revealed few clinically significant changes in vital signs, laboratory tests, and electrocardiogram results. These data provide support for the efficacy and safety of desvenlafaxine in the treatment of major depressive disorder. PMID- 17917553 TI - Evidence for the efficacy of duloxetine in treating mild, moderate, and severe depression. AB - Clinicians need to know whether duloxetine is effective in patients across a broad range of depressive symptoms and depression severity. Data were pooled from nine randomized, double-blind, placebo-controlled studies in major depressive disorder (total N=2227) comparing duloxetine (40-120 mg/day) with placebo for 8-9 weeks. Patients were retrospectively stratified by baseline score on the HAMD17 into mild (< or =19; n=682), moderate (n=1099), or severe (> or =25; n=446) groups. Duloxetine produced significantly greater baseline-to-endpoint mean change than placebo in HAMD17 total score, Maier and retardation subscales, and the Clinical Global Impressions-Severity of Illness scale in all three cohorts. Significant improvement was seen in HAMD17 items 1 (depressed mood), 3 (suicide), 7 (work and activities), and 10 (psychic anxiety) regardless of severity. The HAMD17 anxiety subscale and items 13 (somatic symptoms-general) and 15 (hypochondriasis) showed significant improvement only in moderately and severely ill patients. Significant improvement in the HAMD17 Maier subscale was seen in all groups by week 1. In all three groups, placebo was significantly superior to duloxetine at early visits on HAMD17 item 12 (somatic symptoms-GI). Mildly and severely ill patients exhibited significant reduction in visual analog scale overall pain severity at the study endpoint. The studies contained fewer patients with very mild or very severe illness, limiting our ability to draw conclusions in these patient populations. Duloxetine demonstrated superior efficacy in the treatment of major depressive disorder, when compared with placebo, regardless of the baseline severity of depressive symptoms, although effect sizes were largest in the most severely depressed patients. PMID- 17917554 TI - Risperidone alone versus risperidone plus valproate in the treatment of patients with schizophrenia and hostility. AB - The objective of the study was to compare the antiaggressive efficacy of risperidone monotherapy versus risperidone plus valproate in patients with schizophrenia. This was an 8-week open-label randomized parallel group clinical trial in hospitalized adults diagnosed with schizophrenia and with hostile behavior. Patients were randomly assigned to receive risperidone alone (n=16) or risperidone plus valproate (n=17). To minimize bias, raters were blinded to the assigned treatment arm. Outcome measures included the Positive and Negative Syndrome Scale (PANSS), Buss-Durkee Hostility Inventory (BDHI), Barratt Impulsiveness Scale (BIS), Nurses Observation Scale for Inpatient Evaluation (NOSIE), and the Overt Aggression Scale (OAS). Although significantly fewer patients randomized to monotherapy completed the study (chi(2)=8.62, d.f.=1, P=0.003), no significant differences between monotherapy or combination treatment were observed in change of the BDHI, BIS, NOSIE, PANSS total scores, OAS measures of aggressive behavior or the hostility item of the PANSS. In conclusion, although patients receiving combination treatment were more likely to complete the study, we were unable to detect a meaningful advantage for combination therapy as measured by rating scales. PMID- 17917555 TI - Comparison of ziprasidone and aripiprazole in acutely ill patients with schizophrenia or schizoaffective disorder: a randomized, double-blind, 4-week study. AB - We compared the efficacy and safety of ziprasidone and aripiprazole in the treatment of acutely ill patients with schizophrenia. Patients were randomized to receive double-blind treatment with ziprasidone (80-160 mg/day), or aripiprazole (10-30 mg/day) for up to 4 weeks. Primary efficacy measures were the Clinical Global Impression of Severity scale (CGI-S) and Brief Psychiatric Rating Scale (BPRSd) total (derived from the Positive and Negative Syndrome Scale). Noninferiority for ziprasidone (N=125) relative to aripiprazole (N=128) was established for CGI-S score (P=0.007), but was not confirmed for BPRSd total score (P=0.248). Effect sizes for within-group improvement, however, were robust for both ziprasidone and aripiprazole (effect size range 1.0-1.1 for CGI-S; and range 1.1-1.2 for BPRSd total). A mixed model repeated measures analysis of BPRSd total score favored ziprasidone at day 4 compared with aripiprazole (P=0.04), with no significant differences between treatment groups at other visits (P=0.001 for interaction between treatment and visit). No statistically significant difference was found in CGI-S score between groups across all visits. Our findings suggest that ziprasidone and aripiprazole exhibit similar efficacy and tolerability profiles in the treatment of acute schizophrenia. Differences between the two drugs in the onset of therapeutic effect warrant further investigation. PMID- 17917556 TI - A randomized, single-blind, comparison of venlafaxine with paroxetine in elderly patients suffering from resistant depression. AB - It is estimated that up to 45% of patients with depression do not have an adequate response to a first trial of antidepressant therapy with even higher reported rates for the elderly patients. To compare the efficacy and the tolerability of venlafaxine vs. paroxetine in elderly patients suffering from resistant major depression, who did not respond to at least two previous adequate trials of antidepressants. Patients entered an 8-week single-blind study. Patients were rated using the Clinical Global Impression Scale, Hamilton Rating Scale for Depression, and the Geriatric Depression Scale. Assessments were performed at baseline and on days 7, 14, 21, 28, 42 and 56. Side effects were recorded in a systemic manner. Thirty patients were included in the study, (17 women, 13 men; mean age=75.9 years, range: 68-83) and all had completed the 6 week trial. Mean dose of venlafaxine used was 165 mg/day (SD=73.8; range 75-300 mg). Mean dose of paroxetine used was 26 mg/day (SD=15.04; range 10-60 mg). Nine patients treated with venlafaxine (60%) and five patients treated with paroxetine (33%) remitted after 8 weeks of treatment. Four patients treated with venlafaxine and eight patients treated with paroxetine failed to respond. Significant improvement in Hamilton Rating Scale for Depression scores between baseline and endpoint were observed in both groups of patients. The mean Hamilton Rating Scale for Depression change for paroxetine was -12.5 and for venlafaxine -19.1 (P<0.05). The mean Geriatric Depression Scale change for paroxetine was -3.2 and for venlafaxine -6.0 (P<0.3). The mean Clinical Global Impression Scale change was -2.3 for paroxetine and -3.5 for venlafaxine (P<0.05). Venlafaxine was significantly superior to paroxetine on Clinical Global Impression Scale and Hamilton Rating Scale for Depression measures. Side effects were transient and did not differ between treatment groups. Elderly depressed patients resistant to previous treatments had responded to a trial of paroxetine or venlafaxine. Remission rates were higher for venlafaxine and tolerability was acceptable for both compounds. PMID- 17917557 TI - Quetiapine for the continuation treatment of bipolar depression: naturalistic prospective case series from the Stanley Bipolar Treatment Network. AB - Continuation treatment for bipolar disorder often consists of a mood stabilizer and a second-generation antipsychotic. Quetiapine has been shown to be an effective treatment for acute mania and acute bipolar depression, but there are limited data for its use in continuation treatment. This study examined the effectiveness of open-label adjunctive quetiapine therapy for continuation treatment in patients with bipolar disorder. Prospectively collected life chart data from 63 outpatients with bipolar disorders, most recent episodes depressed, manic, or cycling, who received adjunctive quetiapine therapy as part of standard acute treatment were analyzed. Patients had 4 or more weeks of prequetiapine baseline data and at least 2 weeks of quetiapine treatment with no other medication changes. Patients were grouped by baseline symptoms; depression only, mania only, or both mania and depression (cycling group). Owing to small mania and well groups (n=4), differences between depression and cycling groups were examined and mania and well groups excluded. Fifty-five patients were included in the analyses. The primary outcome measure was change in mood severity from baseline to change in treatment regimen, as measured by the NIMH Life Charting Method. Patients received adjunctive quetiapine for a mean of 122 (SD=149) days. Both groups showed significant improvement in depression ratings and time spent depressed by week 10. Both groups showed significant improvement in overall mood. No between-group differences in improvement were found. Adjunctive quetiapine may be useful as continuation treatment in bipolar populations with both pure depressive and cycling symptoms. Further controlled studies are warranted. PMID- 17917558 TI - No increased incidence of diabetes in antidepressant users. AB - This study investigated whether the association between depression and diabetes was influenced by the presence of chronic somatic disease. To distinguish between depression and other psychosocial complaints, we studied the onset of diabetes in antidepressant (AD) users and benzodiazepine (BD) users, respectively. From the PHARMO database, which includes complete drug prescription data, we identified subjects using (i) no ADs and no BDs; (ii) AD but no BD; (iii) BD but no AD; and (iv) AD and BD. A total of 60 516 individuals (age: 45.5+/-17 years; 42.1% men) were followed from their first prescription for AD or BD until end of registration or a first prescription for antidiabetic drugs. The crude incidence rate in AD but no BD users was not increased compared with no AD and no BD users. After adjustment for age, sex and chronic diseases, the hazard ratios (95% confidence interval) were 1.05 (0.88-1.26) for AD but no BD users, 1.21 (1.02 1.43) for BD but no AD users and 1.37 (1.12-1.68) for AD and BD users compared with no AD and no BD users. We did not find an increased risk of diabetes in individuals using ADs. The association between BD use and diabetes was partly explained by chronic somatic comorbidity. PMID- 17917561 TI - From circadian rhythms to clock genes in depression. AB - Depression, particularly unipolar depression, has often been linked to circadian (i.e. approximately 24-h) rhythm abnormalities. The observation that many diverse rhythms are disrupted in depressed patients indicates that such disturbances are not unique to specific rhythms, but instead are of a more central origin (i.e. involve the central circadian pacemaker and/or the molecular circadian clock core machinery). One rhythm that is often disrupted in depression is the sleep-wake cycle - a disruption that, in turn, might lead to other rhythm disturbances. Thus, there are two general ways in which disrupted circadian rhythms could lead to depression: (i) disorganization within the circadian system could itself lead to neurobiological dysfunction and (ii) a circadian disturbance of the normal sleep-wake cycle could facilitate or exacerbate the depressed state. The recent discovery of the molecular clock responsible for the generation of circadian rhythms provides novel mechanistic insights into how rhythm abnormalities might lead to disrupted behavioural states, and offers new therapeutic avenues for the treatment of the timing abnormalities that might underlie depression. The finding that the molecular circadian clock is present in many cells in the central nervous system and regulates the timing of the expression of at least 10% of the transcripts in many tissues emphasizes how circadian dysfunction could have drastic consequences for normal physiological function in the brain. PMID- 17917562 TI - Evidence of agomelatine's antidepressant efficacy: the key points. AB - Depressive disorders are of the highest socioeconomic and health-economic importance, as they are the psychiatric disorders that most frequently cause psychosocial disability. Despite the progress that has been made, currently available pharmacotherapies for depression still have a limited antidepressant efficacy with a delayed onset of several weeks, and still cause side effects; these unmet needs represent important reasons to continue to search for novel treatment options. A disorganization of circadian rhythms has been suggested to play an important role in the pathophysiology of major depression, and complaints regarding disturbed sleep are frequent in depressed patients. As endogenous melatonin secretion underlies the regulation of circadian rhythms, compounds with activity at melatonergic receptors have been proposed as potential novel therapeutics. Agomelatine (S-20098), a compound with agonistic properties at MT1 and MT2 receptors and antagonistic properties at the 5-HT2C receptor, has been shown preclinically to exhibit robust antidepressant effects in several experimental paradigms. Clinical trials, including phase III studies, have now demonstrated the superior efficacy of agomelatine in comparison with placebo, and a similar efficacy in comparison with active comparators, for the treatment of major depression. Agomelatine was even effective in severely depressed patients. In all studies published so far, agomelatine was found to be safe and its overall tolerability profile was superior to that of selective serotonin reuptake inhibitors and selective serotonin and noradrenaline reuptake inhibitors. PMID- 17917563 TI - High-quality remission: potential benefits of the melatonergic approach for patients with major depressive disorder. AB - Full remission of symptoms is the goal for the acute treatment of depression, because incomplete remission is associated with poor outcomes including higher risk of relapse and chronicity. The current definitions for remission (e.g. a score of left). Neuropathologic examination displayed the highest degree of degeneration and ubiquitin/TDP-43 pathology in the proband's parietal areas. Genetic analysis revealed the presence of the c.26C>A PGRN mutation in 1 allele. This mutation has been reported in association with hereditary-dysphasic disinhibition-dementia, Alzheimer-like dementia, progressive supranuclear palsy, and primary progressive aphasia. The peculiar findings observed in this patient indicate that the parietal lobe may represent the most vulnerable anatomical area in some of the PGRN-associated frontotemporal lobar degeneration with ubiquitin/TDP-43-positive inclusion cases. PMID- 17917581 TI - Location, location, location: altered transcription factor trafficking in neurodegeneration. AB - Neurons may be particularly sensitive to disruptions in transcription factor trafficking. Survival and injury signals must traverse dendrites or axons, in addition to soma, to affect nuclear transcriptional responses. Transcription factors exhibit continued nucleocytoplasmic shuttling; the predominant localization is regulated by binding to anchoring proteins that mask nuclear localization/export signals and/or target the factor for degradation. Two functional groups of karyopherins, importins and exportins, mediate RanGTPase dependent transport through the nuclear pore. A growing number of recent studies, in Alzheimer, Parkinson, and Lewy body diseases, amyotrophic lateral sclerosis, and human immunodeficiency virus encephalitis, implicate aberrant cytoplasmic localization of transcription factors and their regulatory kinases in degenerating neurons. Potential mechanisms include impaired nuclear import, enhanced export, suppression of degradation, and sequestration in protein aggregates or organelles and may reflect unmasking of alternative cytoplasmic functions, both physiologic and pathologic. Some "nuclear" factors also function in mitochondria, and importins are also involved in axonal protein trafficking. Detrimental consequences of a decreased nuclear to cytoplasmic balance include suppression of neuroprotective transcription mediated by cAMP- and electrophile/antioxidant-response elements and gain of toxic cytoplasmic effects. Studying the pathophysiologic mechanisms regulating transcription factor localization should facilitate strategies to bypass deficits and restore adaptive neuroprotective transcriptional responses. PMID- 17917584 TI - Increased APOBEC3G expression is associated with extensive G-to-A hypermutation in viral DNA in rhesus macaque brain during lentiviral infection. AB - APOBEC3G restricts retrovirus replication through inducing guanosine-to-adenosine (G-to-A) hypermutations in viral DNA. Its role in brain "intrinsic immunity" has not been elucidated nor has it been convincingly demonstrated which brain cell compartments produce this defense factor in human immunodeficiency virus (HIV) infection, acquired immunodeficiency syndrome (AIDS), and antiretroviral therapy. Here, we investigated by immunohistochemistry and in situ hybridization the cell specific regulation of APOBEC3G in rhesus macaque brains during infection with simian immunodeficiency virus (SIV) clone deltaB670, a primate model of HIV disease. We found that APOBEC3G protein and mRNA were exclusively expressed by some perivascular macrophages throughout the brain of noninfected and asymptomatic SIV-infected monkeys. Depending on virus burden, APOBEC3G was induced in microglia/macrophage-derived cells and T lymphocytes in late-stage SIV infection. Intracellularly, APOBEC3G was found in the cytoplasm and/or in the nucleus. APOBEC3G-positive cells were in close proximity to SIV gag-positive cells or were SIV-copositive. Induction of APOBEC3G was accompanied by G-to-A hypermutations in the gag and pol regions of retroviral DNA isolated from brain sections of AIDS-symptomatic monkeys. Although brain-directed treatment with antiretroviral 6-chloro-2',3'-dideoxyguanosine suppressed brain SIV burden, encephalitis and reduced cerebral APOBEC3G synthesis hypermutations were still detectable. Upregulation of APOBEC3G may restrict spread of SIV in the brain and thus limit brain damage during lentiviral infection. PMID- 17917585 TI - Vascular endothelial growth factor prevents paralysis and motoneuron death in a rat model of excitotoxic spinal cord neurodegeneration. AB - Vascular endothelial growth factor (VEGF) delays disease onset and progression in transgenic rodent models of familial amyotrophic lateral sclerosis (ALS). Because most cases of ALS are sporadic, it is important to determine whether VEGF can protect motoneurons in a nontransgenic ALS paradigm. We tested this possibility in a new model of chronic excitotoxic spinal neurodegeneration in the rat. Using osmotic minipumps, we continuously infused the glutamate receptor agonist alpha amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) directly in the lumbar spinal cord. The effect of this treatment on motor behavior was assessed with 3 motor performance tests, and neurodegeneration was evaluated by histologic and immunohistochemical analyses. AMPA infusion produced dose-dependent progressive hindlimb motor deficits, reaching complete bilateral paralysis in approximately 10 days, which was correlated with the loss of spinal motoneurons. VEGF administered together with AMPA completely prevented the motor deficits, and the motoneuron death was reduced by more than 75%. Thus, we have developed an in vivo model of progressive spinal motoneuron death due to overactivation of AMPA receptors. The finding that VEGF protected motoneurons from this AMPA receptor mediated excitotoxic death suggests that it may be a therapeutic agent in sporadic ALS. PMID- 17917586 TI - Selective induction of cAMP phosphodiesterase PDE4B2 expression in experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) in Lewis rats is the most widely used animal model for multiple sclerosis. Cyclic adenosine monophosphate (cAMP) has been associated with neuroinflammation. The aim of this study was to investigate the possible involvement of different cAMP-specific phosphodiesterase (PDE) isoenzymes by analyzing their expression in the brain of EAE rats. We found in the brain of EAE animals that there was a dramatic increase in the mRNA expression levels of the PDE4B isozyme detected around blood vessels from the spinal cord to the upper midbrain. There was a single splicing form of the 4 splice variants that are known for PDE4B: PDE4B2, which showed increased expression levels. This overexpression is localized around the blood vessels and parenchyma in infiltrating T cells and macrophages/microglia. These results support the role played by the activation of the PDE4B2 gene in the neuroinflammatory process in EAE rats. PMID- 17917588 TI - Molecular genetic analysis of p53 intratumoral heterogeneity in human astrocytic brain tumors. AB - We investigated genetic heterogeneity of astrocytic gliomas using p53 gene mutations as a marker. Different parts of morphologically heterogeneous astrocytic gliomas were microdissected, and direct DNA sequencing of p53 gene exons 5 through 8 was performed. Thirty-five glioma samples and tumor-adjacent normal-appearing brain tissue from 11 patients were analyzed. Sixteen different p53 gene mutations were found in 7 patients. We found that some tumors were devoid of p53 gene mutations, whereas other tumors carried 1 or often several (up to 3) different mutations. The mutations were present in grade II, III, and IV astrocytic glioma areas. Both severe functionally dead mutants and mutants with remaining transcriptional activity could be observed in the same tumor. We observed that morphologically different parts of a glioma could carry different or similar mutations in the p53 gene and could be either associated or not associated with the locus of heterozygosity at the mutant site. Coexistence of p53 gene mutations and the locus of heterozygosity was common, at least in astrocytomas grade III and in glioblastomas, and also occurred in astrocytoma grade II areas. These results support the notion that intratumoral heterogeneity in brain tumors originates from different molecular defects. Our results are of importance for a further understanding of the molecular mechanisms behind failure to treat glioma patients. PMID- 17917587 TI - Annexin A1 reduces inflammatory reaction and tissue damage through inhibition of phospholipase A2 activation in adult rats following spinal cord injury. AB - Annexin A1 (ANXA1) has been suggested to be a mediator of the anti-inflammatory actions of glucocorticoids and more recently an endogenous neuroprotective agent. In the present study, we investigated the anti-inflammatory and neuroprotective effects of ANXA1 in a model of contusive spinal cord injury (SCI). Here we report that injections of ANXA1 (Ac 2-26) into the acutely injured spinal cord at 2 concentrations (5 and 20 microg) inhibited SCI-induced increases in phospholipase A2 and myeloperoxidase activities. In addition, ANXA1 administration reduced the expression of interleukin-1beta and activated caspase-3 at 24 hours, and glial fibrillary acidic protein at 4 weeks postinjury. Furthermore, ANXA1 administration significantly reversed phospholipase A2-induced spinal cord neuronal death in vitro and reduced tissue damage and increased white matter sparing in vivo, compared to the vehicle-treated controls. Fluorogold retrograde tracing showed that ANXA1 administration protected axons of long descending pathways at 6 weeks post-SCI. ANXA1 administration also significantly increased the number of animals that responded to transcranial magnetic motor-evoked potentials. However, no measurable behavioral improvement was found after these treatments. These results, particularly the improvements obtained in tissue sparing and electrophysiologic measures, suggest a neuroprotective effect of ANXA1. PMID- 17917589 TI - PARK10 candidate RNF11 is expressed by vulnerable neurons and localizes to Lewy bodies in Parkinson disease brain. AB - The PARK10 locus is associated with idiopathic Parkinson disease (PD), but the responsible gene remains to be identified. Genes associated with familial PD, as well as biochemical evidence from sporadic PD and animal models, have implicated components of the ubiquitin-proteasome system in PD pathogenesis. One attractive candidate gene at the PARK10 locus is RING-Finger Protein 11 (RNF11), the deduced amino acid sequence of which predicts a RING-H2 domain common to E3 ubiquitin ligases such as parkin. To facilitate understanding of this protein and its possible role in PD, we characterized the expression and localization of RNF11 in brain. We detected RNF11 transcript and protein and provided the first direct evidence that RNF11 is expressed in brain. Immunohistochemical analysis of RNF11 protein in rat and human brain, using 2 different antibodies, corroborated the mRNA findings. Both antibodies show that RNF11 is restricted to neurons and excluded from white matter. Moreover, RNF11 is expressed by vulnerable neurons of the substantia nigra and sequestered into Lewy bodies in brains of patients with idiopathic PD. Collectively, these findings identify RNF11 as a strong candidate gene at the PARK10 locus and highlight its potential significance in the development of the common form of PD. PMID- 17917599 TI - Setting sail as a travel therapist. PMID- 17917600 TI - Integrating Iyengar yoga into rehabilitation. PMID- 17917601 TI - Age of menarche and the metabolic syndrome in China. AB - OBJECTIVES: In western populations, young age of menarche is associated with increased cardiovascular risk. Little is known about the potential impact of menarche on the metabolic syndrome (as a proxy for cardiovascular risk) in rapidly economically developing populations where age of menarche is falling. We sought to determine the relation between age of menarche and the metabolic syndrome in a rapidly developing Chinese population. METHODS: We carried out a retrospective historical cohort study of 7349 women from the Guangzhou Biobank Cohort Study, China, enrolled in 2003-2004. Cardiovascular risk factors were obtained from physical examination; age of menarche was obtained from self report. The main outcome measure was the metabolic syndrome and its components. RESULTS: Adjusted for age, education, and number of pregnancies, young age of menarche (<12.5 years) compared with age of menarche > or =14.5 years was associated with a higher risk of the metabolic syndrome (odds ratio = 1.49; 95% confidence interval = 1.22-1.82), central obesity (1.35; 1.10-1.65), raised blood pressure (1.34; 1.09-1.65), raised fasting glucose (1.40; 1.15-1.71), and higher triglyceride levels (1.36; 1.12-1.67). Further adjustment by waist circumference attenuated these effects, but the odds ratios remained elevated. CONCLUSIONS: Earlier age of menarche experienced by younger women in China today, now 12.5 years on average in urban populations, may contribute to an increase in the metabolic syndrome and thereby an increase in cardiovascular disease as these women age. These results further highlight the importance of childhood antecedents of adulthood disease. PMID- 17917602 TI - Preterm delivery and later maternal cardiovascular disease risk. AB - BACKGROUND: Women who have delivered a preterm infant are at elevated risk for cardiovascular disease (CVD), but mechanisms for this association are not understood. METHODS: In a cross-sectional study we investigated whether older women with a history of preterm birth (<37 weeks) had a higher prevalence of CVD. Participants were 446 women (mean age 80 years; 47% black) enrolled in the Pittsburgh, PA field center of The Health, Aging and Body Composition Study. Women reported preterm status, birth weight, smoking status, and selected complications for each pregnancy. CVD status was determined by self-report and hospital records. Analysis was limited to first births not explicitly complicated by hypertension or preeclampsia. RESULTS: Women who had delivered a preterm infant (on average 57 years in the past) had a higher prevalence of CVD. After adjustment for race, age, blood pressure, pulse wave velocity, interleukin-6, high-density lipoprotein cholesterol, and statin use, the odds ratio for CVD among women who delivered a preterm infant was 2.85 (95% confidence interval = 1.19-6.85) compared with women who had delivered term infants weighing more than 2500 g. This relationship was not altered by lifetime smoking history. There was evidence of negative confounding by statin use and high-density lipoprotein cholesterol. Among women delivering infants who were both preterm and low birth weight (<2500 g), the odds ratio was 3.31 (1.06-10.37) for CVD compared with women with term, normal weight infants. CONCLUSIONS: These results suggest that vascular and metabolic factors account for some but not all of the increased prevalence of CVD among women many years after a preterm birth. PMID- 17917603 TI - Anthropometric characteristics and risk of uterine leiomyoma. AB - BACKGROUND: The etiology of uterine leiomyoma (or fibroids) is poorly understood. Sex steroid hormones and growth factors have been hypothesized to play a role in their development, and anthropometric characteristics may influence uterine leiomyoma risk. METHODS: We analyzed prospectively-collected data from the Nurses' Health Study II, a cohort of 116,609 female registered nurses age 25 to 42 years at baseline. Incidence of uterine leiomyoma and anthropometric characteristics were assessed every 2 years. We calculated hazard ratios and 95% confidence intervals adjusted for known and suspected risk factors for uterine leiomyoma. RESULTS: Uterine leiomyomas were associated with current body mass index, change in weight since age 18, and waist-to-hip ratio. However, there was little evidence of their association with body mass index at age 18, childhood or adolescent body size, or adult height. CONCLUSION: Body mass and weight gain in adulthood may increase the risk of uterine leiomyoma, but body mass in early life does not. PMID- 17917604 TI - Squamous cell and basal cell carcinoma of the skin in relation to radiation therapy and potential modification of risk by sun exposure. AB - BACKGROUND: Epidemiologic studies consistently find enhanced risk of basal cell carcinoma of the skin among individuals exposed to ionizing radiation, but it is unclear whether the radiation effect occurs for squamous cell carcinoma. It is also not known whether subgroups of individuals are at greater risk, eg, those with radiation sensitivity or high ultraviolet radiation exposure. METHODS: We analyzed data from a case-control study of keratinocyte cancers in New Hampshire. Incident cases diagnosed in 1993-1995 and 1997-2000 were identified through a state-wide skin cancer surveillance system, and controls were identified through the Department of Transportation and Center for Medicare and Medicaid Service Files (n = 1121 basal cell carcinoma cases, 854 squamous cell carcinoma cases, and 1049 controls). RESULTS: We found an association between history of radiation treatment and basal cell carcinoma. The association was especially strong for basal cell carcinomas arising within the radiation treatment field (odds ratio = 2.6; 95% confidence interval = 1.5-4.3), and among those treated with radiation therapy before age 20 (3.4; 1.8-6.4), those whose basal cell carcinomas occurred 40 or more years after radiation treatment (3.2; 1.8-5.8), and those treated with radiation for acne (11; 2.7-49). Similar age and time patterns of risk were observed for squamous cell carcinoma, although generally with smaller odds ratios. For basal cell carcinoma, early exposure to radiation treatment was a risk factor largely among those without a history of severe sunburns, whereas for squamous cell carcinoma, radiation treatment was a risk factor primarily among those with a sun-sensitive skin type (ie, a tendency to sunburn). CONCLUSIONS: Radiation treatment, particularly if experienced before age 20, seems to increase the long-term risk of both basal and squamous cell carcinomas of the skin. These risks may differ by sun exposure or host response to sunlight exposure. PMID- 17917605 TI - Condom use and its association with bacterial vaginosis and bacterial vaginosis associated vaginal microflora. AB - BACKGROUND: Previous studies have been inconsistent with regard to whether condom use is associated with bacterial vaginosis. We evaluated this association using case-crossover analyses. METHODS: A total of 871 women at high risk for sexually transmitted diseases were followed for a median of 3 years. At baseline and at 6, 12, 24, and 36 months thereafter, vaginal swabs were obtained for gram stain diagnosis of bacterial vaginosis, culture of microflora, and DNA amplification of Neisseria gonorrhoeae and Chlamydia trachomatis. Case-crossover analyses using incident and recurrent incident case periods were used to assess the associations among condom use, bacterial vaginosis, and vaginal microflora. RESULTS: Consistent condom use (10 out of 10 sexual encounters) was associated with a decreased frequency of bacterial vaginosis (adjusted odds ratio = 0.55 [95% confidence interval 0.35-0.88]). When we excluded women with intermediate flora, consistent condom use was even more strongly protective against bacterial vaginosis (0.37 [0.20-0.70]). Consistent condom use was similarly protective against carriage of anaerobic gram-negative pigmented rods (0.58 [0.36-0.94]). Results were similar when analyses were repeated to capture only first occurrences of outcomes among women without bacterial vaginosis at baseline, suggesting a protective effect against the acquisition of bacterial vaginosis. CONCLUSIONS: Consistent condom use was associated with a decrease in the risk for bacterial vaginosis and associated vaginal microflora. PMID- 17917606 TI - Perceived racial discrimination and risk of uterine leiomyomata. AB - BACKGROUND: The incidence of uterine leiomyomata (fibroids, myomas) is 2-3 times higher in black women than white women. Black women also report higher levels of racial discrimination. We evaluated the hypothesis that greater exposure to racism increases myoma risk in black women. METHODS: Data were derived from the Black Women's Health Study, a prospective cohort study of US black women age 21 69 years in 1995. In 1997, women reported on "everyday" and "lifetime" experiences of racism. From 1997 through 2003, we followed 22,002 premenopausal women to assess the association between self-reported racism and risk of myomas. Incidence rate ratios (IRRs) and 95% confidence intervals (CIs) were estimated from Cox regression models. RESULTS: During 107,127 person-years of follow-up, 3440 new cases of uterine myomas confirmed by ultrasound (n = 2774) or surgery (n = 666) were reported. All IRRs for "lifetime" and "everyday" experiences of racism were above 1.0. Using a summary variable that averaged the responses from 5 "everyday" racism items, multivariable IRRs comparing quartiles 2, 3, and 4 to quintile 1 (lowest) were 1.16 (95% CI = 1.04-1.29), 1.19 (1.06-1.32), and 1.27 (1.14-1.43), respectively. Multivariable IRRs comparing women who reported 1, 2, or 3 lifetime occurrences of major discrimination (ie, job, housing, or police) relative to those who reported none were 1.04 (0.96-1.13), 1.17 (1.07-1.28), and 1.24 (1.10-1.39), respectively. Results did not vary according to case definition (ultrasound vs. surgery) or health care utilization. Associations were weaker among foreign-born women and among women with higher coping skills. CONCLUSIONS: Perceived racism was associated with an increased risk of uterine myomas in US born black women. PMID- 17917607 TI - Aircraft noise and incidence of hypertension. AB - BACKGROUND: An association between aircraft noise exposure and hypertension prevalence has been suggested but there are no longitudinal studies of this association. Our aim was to investigate the influence of aircraft noise on the incidence of hypertension. METHODS: A cohort of 2754 men in 4 municipalities around Stockholm Arlanda airport was followed between 1992-1994 and 2002-2004. The cohort was based on the Stockholm Diabetes Preventive Program; half of the study subjects had a family history of diabetes. Residential aircraft noise exposure (expressed as time-weighted equal energy and maximal noise levels) was assessed by geographical information systems techniques among those living near the airport. Incident cases of hypertension were identified by physical examinations, including blood pressure measurements, and questionnaires in which subjects reported treatment or diagnosis of hypertension and information on cardiovascular risk factors. Analyses were restricted to 2027 subjects who completed the follow-up examination, were not treated for hypertension, and had a blood pressure below 140/90 mm Hg at enrollment. RESULTS: For subjects exposed to energy-averaged levels above 50 dB(A) the adjusted relative risk for hypertension was 1.19 (95% CI = 1.03-1.37). Maximum aircraft noise levels presented similar results, with a relative risk of 1.20 (1.03-1.40) for those exposed above 70 dB(A). Stronger associations were suggested among older subjects, those with a normal glucose tolerance, nonsmokers, and subjects not annoyed by noise from other sources. CONCLUSION: These findings suggest that long-term aircraft noise exposure may increase the risk for hypertension. PMID- 17917608 TI - Sexuality in Brazilian women aged 40 to 65 years with 11 years or more of formal education: associated factors. AB - OBJECTIVE: To evaluate factors associated with the sexuality of middle-aged women. DESIGN: Cross-sectional, population-based survey using an anonymous self response questionnaire. A total of 276 Brazilian-born women, 40 to 65 years old with at least 11 years of formal education, participated in the study. The evaluation instrument was based on the Short Personal Experiences Questionnaire. Seven components were analyzed: satisfaction in sexual activities, orgasm, intensity of desire, self-classification of sexual life, frequency of arousal, sexual activity, and sexual fantasies. Sociodemographic, clinical, behavioral, and reproductive factors were evaluated. Data were analyzed using the chi and Fisher exact tests and Poisson multiple regression analysis. Prevalence ratios (PRs) and their 95% CIs were calculated. RESULTS: The median sexuality score was 9 (range, 2.45-13.77). Bivariate analysis indicated that being 50 years of age or older; in the menopausal transition or postmenopause; not having a sexual partner; reporting hot flushes, insomnia, depression, nervousness, sedentary lifestyle, arterial hypertension, or urinary incontinence; and poor self perception of health were significantly associated with a below median sexuality score. Multiple regression analysis showed that the prevalence of below median scores was higher in older women (prevalence ratio [PR] = 1.03, 95% CI: 1.01 1.05) and in those with insomnia (PR = 1.46, 95% CI: 1.08-1.96). Having a sexual partner (PR = 0.68, 95% CI: 0.50-0.92) and feeling well (PR = 0.73, 95% CI: 0.57 0.94) was associated with a protective effect against a below median sexuality score. CONCLUSIONS: Older women and those with insomnia were more likely to have a low sexuality score, whereas those with a sexual partner and who felt well were less likely to have a low sexuality score. PMID- 17917609 TI - Older age and markers of inflammation are strong predictors of clinical events in women with asymptomatic carotid lesions. AB - OBJECTIVE: Limited information exists regarding the association between markers of inflammation, such as high-sensitivity C-reactive protein (hs-CRP) and fibrinogen, and adverse events in postmenopausal women with subclinical atherosclerosis. Therefore, we investigated the prognostic impact of traditional risk factors and inflammation on adverse cardiac events in women with asymptomatic carotid lesions. DESIGN: We studied 250 postmenopausal women who were free of cardiovascular disease. Traditional cardiovascular risk factors were investigated, and laboratory analysis included measurement of plasma lipids, fibrinogen, and hs-CRP. The early phases of carotid atherosclerosis were assessed by B-mode ultrasonography. Women were asked about symptoms or a previous history of coronary artery disease and were followed for a period of 5 years. RESULTS: We found that the increment in age (in quintiles) was significantly associated with higher incidence of current smokers (P = 0.0286), hypertension (P = 0.0230), family history of coronary artery disease (P = 0.0216), dyslipidemia (P = 0.0330), and higher levels of fibrinogen (P = 0.0158). Moreover, older women had a higher prevalence of carotid lesions (P < 0.0001). After the follow-up, cardio- and cerebrovascular events were registered in 22% of the women. Using multivariate analysis, we observed that older age (odds ratio [OR], 1.7; 95% CI, 1.3-2.2; P < 0.0001), fibrinogen (OR, 1.6; 95% CI, 1.2-2.0; P < 0.0001), the presence of carotid lesions (OR, 2.0; 95% CI, 1.4-3.0; P = 0.0002), and hs-CRP (OR, 1.3; 95% CI, 1.2-2.0; P = 0.0175) were predictors of adverse events during the follow-up. CONCLUSIONS: Adverse events occurred more frequently in women with higher levels of fibrinogen and hs-CRP. The significance of these results requires confirmation in other studies, but they may have important implications for screening subjects at risk for cardiovascular disease and identifying candidates for anti-inflammatory therapy. PMID- 17917610 TI - Women are periodontally healthier than men, but why don't they have more teeth than men? AB - OBJECTIVE: Periodontal diseases are more prevalent in men than in women. However, in a population-based epidemiological study, we found that, on average, women have fewer teeth than men. The aim of this study was to test different hypotheses that could explain this obvious paradox. DESIGN: In 4,290 randomly selected participants from the normal population (Study of Health in Pomerania), we determined diagnostic periodontal parameters, attachment loss, and number of teeth. Behavioral and environmental risk factors were assessed by interviews and questionnaires. Use of estrogens was assessed, and urinary excretion of collagen cross-links was determined. RESULTS: Multiple regression analyses adjusted for caries and periodontitis revealed that in the women of this population, there is an inverse association between the number of children born and the number of teeth (P < 0.01). This relationship depends on socioeconomic status, bone metabolism, and the use of estrogens. In the group of the youngest (20-40 years), the bone turnover rate is positively related to the number of children born (P < 0.01). In postmenopausal women treated with estrogens, the number of teeth was higher than in men of the same age group. Only in women without hormone treatment were there fewer teeth. CONCLUSION: The apparent paradox of having fewer teeth despite better periodontal health in women compared with men is related to an increased bone turnover rate and socioeconomic conditions such as low education and low social status. Periodontal health is even worse if these factors are combined. PMID- 17917611 TI - Gabapentin for the treatment of menopausal hot flashes: a randomized controlled trial. AB - OBJECTIVE: To compare the effectiveness and tolerability of gabapentin with placebo for the treatment of hot flashes in women who enter menopause naturally. DESIGN: A randomized, double-blind, placebo-controlled trial was conducted across the greater Toronto area between March 2004 and April 2006 in the community and primary care settings. Eligible participants were 200 women in natural menopause, aged 45 to 65 years, having at least 14 hot flashes per week. Study participants were randomized to receive gabapentin 300 mg oral capsules or placebo three times daily for 4 weeks. The primary outcome measure was the mean percentage change from baseline to week 4 in daily hot flash score, determined from participant diaries. Secondary outcome measures included changes in weekly mean hot flash scores and frequencies, quality of life, and adverse events. RESULTS: Of the 197 participants, 193 (98%) completed the study. Analysis was by intention to treat. Hot flash scores decreased by 51% (95% CI: 43%-58%) in the gabapentin group, compared with 26% (95% CI: 18%-35%) on placebo, from baseline to week 4. This twofold improvement was statistically significant (P < 0.001). The Menopause Specific Quality-of-Life vasomotor score decreased by 1.7 (95% CI: 1.3-2.1; P < 0.001) in the gabapentin group. These women reported greater dizziness (18%), unsteadiness (14%), and drowsiness (12%) at week 1 compared with those taking placebo; however, these symptoms improved by week 2 and returned to baseline levels by week 4. CONCLUSIONS: Gabapentin at 900 mg/day is an effective and well tolerated treatment for hot flashes. PMID- 17917612 TI - Breast arterial calcifications (BACs) found on screening mammography and their association with cardiovascular disease. AB - OBJECTIVE: Breast arterial calcifications (BACs) are common but unreported findings on screening mammograms. This study correlated mammographically detected BACs with coronary artery disease (CAD) risk factors and a history of atherosclerotic cardiovascular disease (ASCVD), estimating the relative risk of ASCVD in patients with BACs. DESIGN: Women arriving for breast cancer screening mammography gave their consent to complete a questionnaire and to allow their mammograms to be analyzed independently for the presence of BACs by certified radiologists, who were blinded to the results of the questionnaire. The questionnaire assessed major risk factors for CAD and gathered information on hormone therapy use. RESULTS: Of the 1,919 women with results, 268 were BAC positive, giving a BAC prevalence of 14%. Five cardiovascular risk factors (age, hypertension, hypercholesteremia, diabetes mellitus, and menopause) were significantly more prevalent in the BAC-positive population (P < 0.001). The BAC positive group also had a significantly higher (P < 0.001) occurrence of ASCVD events (angina, previous myocardial infarction, previous abnormal angiography, previous stroke, and previous coronary artery bypass graft). Multiple logistic regression analysis found BACs to be strongly associated with ASCVD events (odds ratio = 2.29, 95% CI: 1.40-3.74) as compared with other CAD risk factors (including hypertension, cigarette smoking, diabetes mellitus, age, and family history of ASCVD). The association of BAC with ASCVD was present even after accounting for age. CONCLUSIONS: BACs are associated with an increased prevalence of both cardiovascular risk factors and cardiovascular morbidity. BACs may be a practical tool to use as a risk indicator for CAD in women. PMID- 17917613 TI - Effects of hormone therapy and dietary soy on myocardial ischemia/reperfusion injury in ovariectomized atherosclerotic monkeys. AB - OBJECTIVE: Hormone therapy (HT) and dietary soy (Soy) inhibit myocardial ischemia/reperfusion (I/R) injury in nonatherosclerotic animals. The aim of this study was to determine their independent and interactive effects on I/R in monkeys previously fed an atherogenic diet for 15 months. DESIGN: Ovariectomized atherosclerotic monkeys (n = 40) were divided into one of four dietary treatment groups: (1) casein as the protein source, (2) casein and added HT (the equivalent of 5 mug ethinyl estradiol + 1 mg norethindrone acetate daily), (3) Soy protein providing 141 mg total isoflavones daily, or (4) Soy + HT. After 12 months monkeys were anesthetized, and their left anterior descending coronary artery was occluded for 1 hour and reperfused for 4 hours. Infarct size was the percentage of the area at risk not staining with triphenyltetrazolium chloride. Additional measures were myocardial blood flow, stroke volume, coronary output, myeloperoxidase, and malondialdehyde. RESULTS: There was an interactive negative effect of HT + Soy to increase infarct size from approximately 30% (in other groups) to 55% (P = 0.0004). Additionally, there were negative main effects of Soy on blood flow, coronary output, and stroke volume during I/R (all P values <0.05). There were no effects of treatment on either myeloperoxidase or malondialdehyde. CONCLUSIONS: Neither HT nor Soy had beneficial effects, whereas their combination had harmful effects, on myocardial I/R injury in monkeys with preexisting atherosclerosis. The mechanism of this negative interaction remains unclear but may relate to Soy's negative effects on hemodynamics. PMID- 17917615 TI - What about this pill that I found at my pharmacy? PMID- 17917614 TI - Sexual health consequences of premature ovarian failure. PMID- 17917616 TI - The evolving algorithm concerning higher-risk PCI (TEACH-PCI). Introduction. PMID- 17917617 TI - Implications of bleeding and blood transfusion in percutaneous coronary intervention. AB - For patients undergoing percutaneous coronary intervention (PCI), bleeding complications are a major clinical concern. With advances in pharmacotherapy and devices over the past 2 decades, the risk of ischemic outcomes, such as myocardial infarction or death, has decreased. Bleeding complications have more recently become a clinical and research priority. Determining the incidence of and risk factors for bleeding is complicated by the multiple systems used to classify bleeding severity and report bleeding events. The origin of the data, clinical trials versus registries, also influences the incidence of reported bleeding events. Registry data suggest that risk of bleeding among patients undergoing PCI is higher in clinical practice than the incidence observed in clinical trials. Another clinical concern is the possible association between PCI related bleeding complications and myocardial infarction, stroke, or death. Reduction in bleeding risk is a desirable goal that may potentially improve survival and increase comfort for patients undergoing PCI. Using strategies such as careful vascular access, alternative radial artery access, and modified antithrombotic regimen may reduce bleeding during PCI as well as improve patient outcomes. PMID- 17917618 TI - Clopidogrel: who, when, and how? AB - Clopidogrel has demonstrated improved outcomes for patients with acute coronary syndromes in several large randomized controlled trials. However, some questions exist about the use of clopidogrel in practice. Who benefits from clopidogrel? When should clopidogrel treatment be initiated? How much clopidogrel should be administered and for how long? Reviewing the results from trials completed to date that have assessed clopidogrel in patients with acute coronary syndromes may help to answer some of these questions. Clinical trial results have demonstrated a reduction in the composite endpoint of death, myocardial infarction, or stroke for patients with acute coronary syndromes who received clopidogrel plus aspirin compared with aspirin alone. For this patient population, early treatment with clopidogrel more than 6 hours before percutaneous coronary intervention (PCI) was associated with a reduction in the risk of death or recurrent ischemic events. The benefits of initiating patients on a 600-mg loading dose of clopidogrel before PCI have been demonstrated in several clinical trials. Clinical trial results and current guidelines recommend long-term treatment with clopidogrel for up to 1 year after PCI. PMID- 17917619 TI - Risk stratifying the acute coronary syndrome patient: a focus on treatable risk. AB - Providing the optimal treatment for patients who present to the emergency room with chest pains or suspected acute coronary syndrome (ACS) remains a dilemma for many practitioners due to subjectivity, delayed diagnoses, and widely variable mechanisms with similar clinical presentations. In treating patients with chest pain but no obvious electrocardiogram changes, practitioners frequently utilize the American College of Cardiology (ACC)/American Heart Association (AHA) Guidelines. The guidelines group possible ACS patients together as unstable angina/non-ST-segment elevation myocardial infarction (NSTEMI) and recommend that treatment be based on level of risk. The challenge for practitioners is discriminating between "risk" and "treatable risk." Evaluation of troponin levels can help identify patients with possible ACS who are at high risk of death and MI, and guide early decision making. Available data indicate that in the troponin negative patient, routine interventions such as unfractionated heparin, glycoprotein IIb/IIIa receptor antagonists, and invasive approaches have no benefit in terms of reducing death and MI. Although the ACC/AHA Guidelines combine patients with unstable angina and NSTEMI, it is essential to evaluate troponin status in order to optimize patient outcomes and safety in the treatment of suspected ACS. PMID- 17917620 TI - Coronary intervention in patients with diabetes, chronic renal disease, and the elderly: therapeutic implications. AB - Patients with diabetes mellitus, chronic renal disease, or advanced age who are undergoing percutaneous coronary intervention are at an increased risk of bleeding and thrombosis. This article reviews the clinical implications of these conditions and discusses the therapeutic options currently available for these patient groups. PMID- 17917622 TI - Road traffic accident-bangladesh stance and proposed action plan. PMID- 17917621 TI - Anticoagulation for acute coronary syndromes: from heparin to direct thrombin inhibitors. AB - The anticoagulant properties of heparin were discovered in 1916, and by the 1930s researchers were evaluating its therapeutic use in clinical trials. Treatment of unstable angina with unfractionated heparin (UFH), in addition to aspirin, was introduced into clinical practice in the early 1980s. UFH was combined with aspirin to suppress thrombin propagation and fibrin formation in patients presenting with acute coronary syndromes (ACS) or patients undergoing percutaneous coronary intervention (PCI). However, UFH stimulates platelets, leading to both activation and aggregation, which may further promote clot formation. Clinical trials have demonstrated that newer agents, such as the low molecular-weight heparins (LMWHs), are superior to UFH for medical management of unstable angina or non-ST-segment elevation myocardial infarction. Increasingly, the LMWHs have been used as the anticoagulant of choice for patients presenting with ACS. For patients undergoing PCI, LMWH provides no sub-stantial benefit over UFH for anticoagulation; however, direct thrombin inhibitors (DTIs) have demonstrated safety and efficacy in this setting. UFH is likely to be replaced by more effective and safer antithrombin agents, such as DTIs. DTIs have antiplatelet effects, anticoagulant action, and most do not bind to plasma proteins, thereby providing a more consistent dose-response effect than UFH. The FDA has approved 4 parenteral DTIs for various indications: lepirudin, argatroban, bivalirudin, and desirudin. The antiplatelet, anticoagulant, and pharmacokinetic properties of bivalirudin support its use as the anticoagulant of choice for both lower- and higher-risk patients, including those undergoing PCI. PMID- 17917623 TI - Childhood acute poisoning in a tertiary medical college hospital of Bangladesh. AB - A total of 193(4.7%) cases of acute childhood poisoning admitted to paediatric Unit of Khulna Medical College Hospital, Bangladesh over a period of two years were studied to see the age and sex distribution, ingredients used, seasonal variation etc. Out of them 107(55.4%) cases were male and rest 86(44.6%) were female. 01-03 years was the most vulnerable age group to be affected. Kerosene was the commonest form of ingredient to be used. Overall mortality rate was 4.66%. Childhood poisoning was a common Medical emergency with considerable morbidity and mortality. Parents should be educated to keep the kerosene out of reach of the children. So, that mortality and morbidity can be reduced to a great extent. PMID- 17917624 TI - Salmonella-a new threat to neonates. AB - Hospital admitted 582 neonates with suspected septicaemia were studied in Microbiology Department of Dhaka Medical College, Dhaka, Bangladesh. Samples being taken from neonatal unit of Dhaka Medical College Hospital and a Neonatal Private Hospital in one year (January to December 2004). Blood culture was done by Lytic-centrifugation method. The isolated organisms were identified using standard laboratory procedures. Among 582 sick neonates 59(10.14%) were culture positive and the predominant organisms were Gram Negative Bacteria (89.83%). Among the isolates Klebsiella spp. was the prime organism (33.90%). Salmonella was observed as the 2nd most common cause (22.03%) for sepsis of neonates. Of these isolated Salmonella strains 46.15% were Salmonella typhi and 53.85% were Salmonella spp. Next to Imipenem, Ciprofloxacin was observed as the drug of choice for treatment of Sepsis neonatorum. PMID- 17917625 TI - Efficacy of combination therapy for the management regime of Alopecia areata. AB - Alopecia areata is a world wide cosmetic problem. Cases are seen in Bangladesh. Due to financial constraint and lack of modern facilities the patients of this country needs cheap, easily available and conveniently usable treatment modality. The present study revealed that the oral administration of prednisolone for 06 weeks followed by topical application of 02% minoxidil for another period of 14 weeks is noninvasive and could act as better remedy for alopecia areata. It is claimed that steroid could induce hair growth through immunosuppression rather than a direct effect on hair growth promotion. When applied together steroid it induced regrowing hair gave more encouraging result. The combination therapy indicated that the gradual loss of hair could be minimized and regrowth of hair could be made possible. To fulfill the demand for the best compliance of treatment modality further study should be directed and dictated amongst a large group of population. PMID- 17917626 TI - Prevalence of psychiatric ailments among patients with sexually transmitted disease. AB - A cross-sectional descriptive study conducted on 250 sexually transmitted disease patients was carried out in two teaching institutes and their tertiary hospitals. These subjects constitute a special group of population for psychiatric diagnosis by using Structured Clinical Interview for Diagnostic and Statistical Manual IIIR. The study was done in the dermatology and venereology outpatient department of two tertiary hospitals of Dhaka, between January 1998 and January 1999. The findings show that 34% of total sexually transmitted disease patients had psychiatric disorders. Anxiety disorders (11.2%) were found to be the most common disorder among these psychiatric patients; this was followed by depressive disorder (8.4%), psychoactive substance use disorder (6.8%), sexual dysfunction (6.8%), bipolar mood disorder (0.4%), and schizophrenia (0.4%). Four percent of anxiety disorder was associated with psychoactive substance use disorder and sexual dysfunction. Similarly 3.6% of depressive disorder was found with psychoactive substance use disorder and sexual dysfunction while 1.6% of sexual dysfunction was associated with substance use disorder. Most of the patients in the sexually transmitted disease population recruited in the present study had both anxiety disorder and depressive disorder though majority of them were undetected and untreated. These findings underscore that special attention needs to be given to the mental health component of our health care delivery system. PMID- 17917627 TI - Pattern of cancer in young adults at National Institute of Cancer Research and Hospital (NICRH), Bangladesh. AB - This is an analysis of 1624 young adults (20-44 years) new cancer patients who attended Out Patient Department of NICRH in 2005 from 01 January to 31 December. This study was carried out to know the age and sex distribution and types of cancer they are suffering from. Histologiacally or cytologically confirmed patients or those patients having radiological or clinical evidence of malignancy were included in the study. There were 42.9% male and 57.1% female with male to female ratio of 0.75: 1.00. Female populations of this age group were the most cancer sufferers. Mean age of young adults was 34.47 (SD+/-6.33) years. Muslims (93.4%) were in majority and 36.5% were illiterate. This study revealed that the top five cancers found in both sexes were breast cancer (19.2%), female genital organ cancer (17.7%), GIT cancer (13.5%), head & neck cancer (12.2%) and respiratory tract cancer (7.9%). Breast cancer and GIT cancer ranked top among females and males respectively. PMID- 17917628 TI - Serum HDL-cholesterol in women using low dose oral contraceptives. AB - This study was done to appraise the effects of low-dose oral contraceptives (OCs) containing 30microgm ethinyl estradiol and 150microgm levonorgestrel on lipid metabolism by a lipid variable-serum HDL-cholesterol (HDL-C). HDL-C has been identified as a lipoprotein that protects against atherosclerosis and its levels are inversely proportional to the risk for developing ischemic coronary disease and venous thromboembolic disorders. Ninety young women within reproductive age group were picked for this study. Sixty women using low-dose oral contraceptives served as experimental group and thirty age matched hormonal contraceptive non users were selected for control group. Experimental group was again subdivided into OCs users for last one-year group, three-year group and five-year group. The result showed that there was no significant difference on serum HDL-cholesterol levels between users & non-users women. It is concluded that low-dose oral contraceptives regimens did not significantly affect the lipid metabolism. It can be safely used as contraceptive & non-contraceptive purposes. The value of studied parameter for serial longer duration of OCs uses needs to be assessed. PMID- 17917629 TI - Impact of etiology of chronic renal failure on growth in children. AB - A prospective study was carried out in the Department of Pediatrics, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh, from October, 2001 to October, 2003 to find out the impact of different etiology of chronic renal failure on growth in children. Fifty children of both sexes under 15 years of age with clinical and biochemical evidence of chronic renal failure (CRF) with creatinine clearance (Ccr) of <75 ml/min/1.73m2 were included in the study. On the basis of underlying causes of CRF, the children were divided into congenital (n=30) and acquired (n=20) groups. All patients' height, weight, radiographs of different bones was obtained to evaluate the presence of renal osteodystrophy (ROD) and for assessment of bone age. Serum intact parathormone (iPTH) level was also assayed in all patients. These parameters were evaluated in two groups. CRF children due to congenital anomalies had stunting and wasting in 23 (76.7%) and 20 (66.7%) cases respectively and the difference between two groups was highly significant (P<0.001). Alkaline phosphatase (467.70+/-218.55 U/L) and iPTH (91.43+/-33.42 pg/ml) were also significantly higher in the congenital group (P<0.001 and P<0.05 respectively). Radiographic features of ROD were present in 15 (50%) cases in congenital group in comparison to 4 (20%) in acquired group and the growth zone lesion was the commonest type of ROD in congenital group (66.7%). CRF should be diagnosed as early as possible to maintain growth potential. PMID- 17917630 TI - Oral contraceptives in gall stone diseases. AB - A cross sectional study was under taken to evaluate the prevalence of gallstone in fertile women who are taking oral contraceptives. The aim of study was to determine any significant association between oral contractive use and Cholelithiasis. Total number of patient was 340. A standard written questioner, which is dully filled by concerned doctor and examination done. History of use of oral contraceptive pills, duration of use and time of taking oral contraceptives drugs whether before, in between pregnancy or after family completion is recorded. Total number of patient was 340 of them 186 patient taken contraceptives and 154 patients without contraceptives. Incidence of gallstone shows that with contraceptives the 21-30 years age group 72(39.13%) without contraceptives 22(14.28%), in 31-40 years age group with contraceptives 74 (40%), without contraceptives 28(18.8%) where as in 41-50 years age group with contraceptives 37(20.10%) and without contraceptives 44(28.57%), 51 years and above age group with contraceptives is 01(0.54%) and without contraceptives is 60(38.96%). Significantly higher incidence of gallstones found in younger patient taking oral contraceptives than without contraceptives, but in older age group incidence is more in-patients without contraceptives than with contraceptives. Oral contraceptives increase the incidence of gallstones disease in younger women especially in early part of their use of oral contraceptives. PMID- 17917632 TI - Verrucous carcinoma of vulva. AB - Vulval carcinoma is an uncommon disease of old women and may manifest as verrucous carcinoma. Intraepithelial cancer of the vulna of women in their twenties and thirties is increasing. Invasive carcinoma of vulva usually appears long time after intraepithelial neoplasia or chronic irritation. We are presenting one young lady of 21 years who noticed whitening of vulva since her childhood and irregular vulval itching in that area. After nine to ten years she further noticed swelling in that area which used to discharge off an on along with itching. As she got married and had disturbed sexual life, on consulting with a specialist doctor, the lesion was diagnosed as an ulcerated malignant growth. Further investigation revealed the lesion to be as verrucous carcinoma of vulva. This is the first report of such case in our country. Any persistent complaints of vulva like itching, discharge, even ulcer should be taken care of for detection of intraepithelial vulvar cancer. PMID- 17917631 TI - Outcome of management of differentiated thyroid carcinoma. AB - A prospective study was done on 93 cases of differentiated thyroid carcinoma to find out the outcome of management according to the protocol followed in the Department of ENT-Head and Neck Surgery of Bangabandhu Sheikh Mujib Medical University Hospital. Here Papillary carcinoma is more common (76.35%) than follicular carcinoma. Papillary carcinoma affected in the younger patients more commonly than the follicular carcinoma with a mean age of 34.37years (SD=12.81) for papillary carcinoma and 44.93 years (SD=16.01) for follicular carcinoma; but age as a risk factor showed no significant difference between two histological types (p>0.05). Female was the predominant sex with a ratio of 3.04:1. In this series majority of patients were categorized as high risk group (75.27%). There is strong association between histological type & risk group (P<0.05). High risk group is more common in follicular carcinoma (95.45% Vs 69.01%). There was a significant differences between the two histological types for extrathyroidal extention and distant metastasis as risk factors (p<0.01 and p<0.001); but tumour size and lymphatic metastasis showed no significant difference (p>0.05). All low risk patients were treated by hemithyriodectomy followed by life long thyroxine. All high risk cases were treated with total thyroidectomy with (35.71%) or without (62.86%) different types of neck dissection (according to the degree of lymph node involvement) followed by radioiodine ablasion and lifelong thyroxine therapy routinely. Overall rate of complication of thyroid surgery was 23.08%. The commoner complications were vocal cord palsy (5.5%), hypoparathyroidism (5.5%) and haemorrhage (4.4%). Less common complications were laryngeal oedema (2.2%), wound infection (2.2%), tracheal injury (1.1%) and death (2.2%). Many cases lost to follow up. Sixty seven cases (73.64%) responded well for follow-up. Mean duration of follow up was 3.40 years (SD=1.41). There was recurrence in thyroid remnant in 9.09% of hemithyroidectomy cases and were treated by completion thyroidectomy followed by radioiodine ablasion. Regional lymphatic metastasis was seen in six cases and was treated by different types of neck dissection. Recurrence at distal site was seen in 4 cases (18.18% of follicular carcinoma). These were treated by radioiodine ablation; two of them died during the period of follow-up. PMID- 17917633 TI - A giant vesical calculus. AB - Massive or giant vesical calculus is a rare entity in the recent urological practice. Males are affected more than the females. Vesical calculi are usually secondary to bladder outlet obstruction. These patients present with recurrent urinary tract infection, haematuria or with retention of urine. We report a young male patient who presented with defaecatory problems along with other urinary symptoms. The patient having an average built, non diabetic but hypertensive. The stone could be palpated by physical examination. His urea levels were within normal limits but urine examination shows infection. USG reveals bilateral hydronephrosis with multiple stones in both kidneys along with a giant vesical calculus. After controlling urinary infection and hypertention he underwent an open cystolithotomy. During operation digital rectal help was needed to remove the stone as it was adherent with bladder mucosa. Post operative period was uneventful. His urinary output was quite normal and had no defaecatory problems. Patient left the hospital 10 days after operation. PMID- 17917634 TI - Vaginal bleeding with multicystic ovaries and a pituitary mass in a child with severe hypothyroidism. AB - A seven year and ten months old girl presented with cyclic vaginal bleeding and a huge abdominopelvic mass. She had clinical features of hypothyroidism. The investigation results were consistent with the diagnosis of primary hypothyroidism with precocious puberty. She also had bilaterally enlarged cystic ovaries on CT scan of abdomen and CT scan of brain showed pituitary macroadenoma. After starting treatment with thyroxine, patient became euthyroid and her general condition improved. Treatment with thyroxine alone halted the cyclic vaginal bleeding, led to rapid resolution of the ovarian cysts and regression of the pituitary mass. PMID- 17917635 TI - Papillon-lefevre syndrome with congenital hepatic fibrosis. AB - Papillon Lefevre syndrome (PLS) is a rare autosomal recessive disorder, which is characterized by palmar-plantar hyperkeratosis, periodontitis, and premature loss of dentition. We report a 16 years old girl with PLS. The patient presented at 08 years of age with complaints of corn on the feet and hands, and failure to thrive. On examination, her upper primarily canines were loose, she had severe periodontitis, eruption of permanent teeth, diffuse eritematous and hyperkeratotic palms and soles that suggested the syndrome. During the follow-up, the patient was diagnosed to have congenital hepatic fibrosis (CHF) when she was 16 years old, while she was being investigated for the etiology of her splenomegaly and pancytopenia. We report a patient with PLS associated with CHF, an association that has not been previously described. Abbreviations-HbsAg: Hepatitis B virus surface antigen, Anti Hbs: Antibody against Hepatitis B surface antigen, Anti Hbc IgM: Antibody against Hepatitis B cor antigen immunglobulin M, Anti dsDNA: Antibody against double stranded deoksiribonucleic acid, Anti HCV: Antibody against Hepatit C virus, Anti HIV: Antibody against human immun deficiency virus, AST: Aspartat amino transferase, ALT: Alanin amino transferase, Gamma-GT: Gamma glutamyl transferase, LDH: Lactate dehydrogenase & MRI: Magnetic resonance imaging. PMID- 17917636 TI - Planned conception in a hyper immunized rhesus-d negative mother by elective plasmapheresis. AB - Here we report a case of planned pregnancy in a Hyper immunized Rh D negative Banker lady who was interested to have a healthy baby in her fourth Gestation as previous conceptions were ended by intra uterine death (IUDs) due to Rhesus Hemolytic diseases of new born (HDN) which is a condition where the lifespan of the infant's red cells is shortened by the action of specific antibodies derived from the mother by placental transfer. The disease begins in intrauterine life and is therefore correctly described as hemolytic disease of the fetus (HDF) and new born, but the simple term HDN has been used for a long time and can be taken to include hemolytic disease of the fetus (HDF). This hemolytic process takes place in utero and results in marked compensatory overproduction of young nucleated red cells in fetal erythropoietic sites. For this reason the disease also called erythroblastosis foetalis. Elective plasmapheresis done at the Transfusion Medicine Department of BSMMU, Dhaka on her, 800 ml. plasma were extracted in 4 different sessions during her antenatal period. One healthy male baby was delivered by LUCS at 32 weeks of pregnancy; the Baby has to receive 170 ml. O negative fresh Whole Blood as Exchange Transfusion to correct mild hyper bilirubinimia and anemia. Manual plasmapheresis may thus be practiced to all Hyper-immunized carrying mothers to prevent intra uterine death (IUD) in Rhesus D negative carrying mothers. PMID- 17917637 TI - Clinico-laboratory profile of 45 filarial arthritis cases. AB - This prospective observational study was done during the period from January 2000 to December 2004 including 45 cases of lymphatic filariasis manifested with acute arthritis. Different investigations were carried out to exclude allergy, parasitic and infectious diseases, autoimmune disorder and malignancy. They were given standard treatment with oral non-steroidal anti-inflammatory drugs (NSAIDs). Articular symptoms were not relived satisfactorily. Later they were given oral corticosteroids. During treatment slight relief of symptoms were noted, but all sign-symptoms reappear after withdrawal of corticosteroid drugs. Treatment with oral diethylcarbamazine citrate (DEC) (150 mg/day) for 3 weeks showed complete resolution of arthritis. No side effects or relapse were encountered. The basic mechanism of relief of pain remains unknown. It was assumed that during benign course of lymphatic filariasis, development of arthritis result most likely due to reaction against some occult agents in the joints. PMID- 17917638 TI - Carcinoma-rectum in an 11 years old boy. AB - Carcinoma rectum is a very uncommon malignancy in childhood. The incidence of colon cancer is found to be 1.3 cases per million children. An eleven years old boy with carcinoma rectum was admitted in the Department of Paediatric Gastroenterology, Bangabandhu Sheikh Mujib Medical University (BSMMU). The boy presented with irregular fever, altered bowel habit, poor appetite, gradual weight loss and abdominal pain. His abdomen was tender with no organomegaly. The rectal wall was tender and hard mass could be felt in the surrounding area on direct rectal examination. Diagnosis was confirmed by biopsy of the tissue from the growth. Surgical resection and anastomosis was done. After one month of surgery chemotherapy was started. Since carcinoma rectum in children has not been reported in Bangladesh we share the information with professionals. PMID- 17917639 TI - Ticket scalpers. PMID- 17917640 TI - Amidst doubts, space research program takes flight. PMID- 17917641 TI - Karolinska Institute under fire for controversial cancer report. PMID- 17917642 TI - Poor trial design leaves gene therapy death a mystery. PMID- 17917643 TI - Straight talk from... Colin Blakemore. PMID- 17917645 TI - pH paper trumps expensive kits in measuring acidity. PMID- 17917646 TI - Microscopes made from bamboo bring biology into focus. PMID- 17917647 TI - Teachers' group brings genomics revolution to minority colleges. PMID- 17917649 TI - Turning one lab's trash into another's treasure. PMID- 17917650 TI - Simpler tests for immune cells could transform AIDS care in Africa. PMID- 17917651 TI - Straight talk from... Eva Harris. PMID- 17917652 TI - Reply to 'UK set to reverse stance on research with chimeras'. PMID- 17917654 TI - It's what you do with the fat that matters! PMID- 17917655 TI - Keeping quiet: microRNAs in HIV-1 latency. PMID- 17917656 TI - Merging mitochondria for neuronal survival. PMID- 17917657 TI - Not so fast: adaptive suppression of innate immunity. PMID- 17917658 TI - Rechallenging immunological memory. PMID- 17917659 TI - Where lies the blame for resistance--tumor or host? PMID- 17917660 TI - 'Multipurpose oxidase' in atherogenesis. PMID- 17917661 TI - The phantom menace? PMID- 17917663 TI - Creation and revelation: two different routes to advancement in the biomedical sciences. PMID- 17917664 TI - Lasker Basic Medical Research Award. Dendritic cells: versatile controllers of the immune system. PMID- 17917665 TI - Lasker Clinical Medical Research Award. The artificial heart valve. PMID- 17917666 TI - Lasker Clinical Research Award. The surprising rise of nonthrombogenic valvular surgery. PMID- 17917667 TI - Lasker Public Service Award. The expanding global health agenda: a welcome development. PMID- 17917669 TI - Target deconvolution strategies in drug discovery. AB - Recognition of some of the limitations of target-based drug discovery has recently led to the renaissance of a more holistic approach in which complex biological systems are investigated for phenotypic changes upon exposure to small molecules. The subsequent identification of the molecular targets that underlie an observed phenotypic response--termed target deconvolution--is an important aspect of current drug discovery, as knowledge of the molecular targets will greatly aid drug development. Here, the broad panel of experimental strategies that can be applied to target deconvolution is critically reviewed. PMID- 17917670 TI - Diagnostic and prognostic markers for gastrointestinal stromal tumors in Norway. AB - Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor in the gastrointestinal tract. The diagnosis of GIST is based on histology together with a panel of immunohistochemical markers; the most important is KIT (CD117). A total of 434 cases of GISTs were confirmed by histology and immunohistochemistry, and incorporated into tissue microarrays. Validation of histological features as well as the prognostic value of two immunohistochemical biomarkers (p16 and L1) was assessed. High mitotic rate, large tumor size, nuclear atypia, and small bowel primary site were all validated as negative prognostic factors in GISTs. Expression of p16 was significantly correlated with unfavorable prognosis, whereas L1 expression was not. PMID- 17917671 TI - Phosphohistone H3 expression has much stronger prognostic value than classical prognosticators in invasive lymph node-negative breast cancer patients less than 55 years of age. AB - The proliferation factor mitotic activity index is the strongest prognostic factor in early breast cancer, but it may lack reproducibility. We analyzed the prognostic value of phosphohistone H3, a marker of cells in late G(2) and M phase, measuring highly standardized immunohistochemical nuclear phosphohistone H3 expression by subjective counts and digital image analysis. Expression was compared with classical clinico-pathologic prognostic variables and the mitotic activity index in 119 node-negative invasive breast cancers in patients less than 55 years old treated with adjuvant systemic chemotherapy with long-term follow-up (median 168 months). Nineteen patients (16%) developed distant metastases and 16 (13%) died. Strong phosphohistone H3 expression occurred preferentially in the peripheral growing front; counts were highly reproducible between observers (R=0.92) and highly consistent with digital image analysis (R=0.96). Phosphohistone H3 correlated (P<0.05) with tumor diameter, estrogen receptor, carcinoma grade, and mitotic activity index. Phosphohistone H3 values were systematically (80%) higher than the mitotic activity index. Receiver-operating curve analysis objectively showed that phosphohistone H3 <13 (n=53; 45% of all cases) vs phosphohistone H3> or =13 (n=66; 55% of all cases) was the strongest prognostic threshold, with 20-year recurrence-free survival of distant metastases of 96 and 58%, respectively (P=0.0002, HR=9.6). Mitotic activity index was the second strongest prognostic variable (P=0.003, HR=3.9). In multivariate analysis, phosphohistone H3 <13 vs> or =13 exceeded the prognostic value of the mitotic activity index. None of the other classical prognostic factors examined offered prognostic value additional to phosphohistone H3. Phosphohistone H3 is by far the strongest prognostic variable in early invasive node-negative breast cancer patients less than 55 years old with long-term follow-up. PMID- 17917672 TI - The origin and application of experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is a model of the neuroimmune system responding to priming with central nervous system (CNS)-restricted antigens. It is an excellent model of post-vaccinal encephalitis and a useful model of many aspects of multiple sclerosis. EAE has been established in numerous species and is induced by priming with a large number of CNS-derived antigens. As a consequence, the pathogenesis, pathology and clinical signs vary significantly between experimental protocols. As I describe in this Timeline article, the reductionist approach taken in some lines of investigation of EAE resulted in a reliance on results obtained under a narrow range of conditions. Although such studies made important contributions to our molecular understanding of inflammation, T-cell activation, and MHC restriction, they did not advance as effectively our knowledge of the polyantigenic responses that usually occur in CNS immunopathology and autoimmunity. PMID- 17917673 TI - Laser scanning cytometry: understanding the immune system in situ. AB - Flow cytometry allows quantitative analysis of the identity and effector function of individual cells. However, it cannot provide information on cellular responses that occur within physiological tissue microenvironments. Laser scanning cytometry is an emerging technology that allows imaging and quantitative analysis of individual cells in tissues in situ. This article describes the technology and its potential for delineating the molecular and cellular events underpinning the immune response in health and disease. PMID- 17917674 TI - Tel1 kinase and subtelomere-bound Tbf1 mediate preferential elongation of short telomeres by telomerase in yeast. AB - Telomerase enables telomere length homeostasis, exhibiting increasing preference for telomeres as their lengths decline. This regulation involves telomere repeat bound Rap1, which provides a length-dependent negative feedback mechanism, and the Tel1 and Mec1 kinases, which are positive regulators of telomere length. By analysing telomere elongation of wild-type chromosome ends at single-molecule resolution, we show that in tel1Delta cells the overall frequency of elongation decreases considerably, explaining their short telomere phenotype. At an artificial telomere lacking a subtelomeric region, telomere elongation no longer increases with telomere shortening in tel1Delta cells. By contrast, a natural telomere, containing subtelomeric sequence, retains a preference for the elongation of short telomeres. Tethering of the subtelomere binding protein Tbf1 to the artificial telomere in tel1Delta cells restored preferential telomerase action at short telomeres; thus, Tbf1 might function in parallel to Tel1, which has a crucial role in a TG-repeat-controlled pathway for the activation of telomerase at short telomeres. PMID- 17917675 TI - The carboxy-terminal coiled-coil of the RNA polymerase beta'-subunit is the main binding site for Gre factors. AB - Bacterial Gre transcript cleavage factors stimulate the intrinsic endonucleolytic activity of RNA polymerase (RNAP) to rescue stalled transcription complexes. They bind to RNAP and extend their coiled-coil (CC) domains to the catalytic centre through the secondary channel. Three existing models for the Gre-RNAP complex postulate congruent mechanisms of Gre-assisted catalysis, while offering conflicting views of the Gre-RNAP interactions. Here, we report the GreB structure of Escherichia coli. The GreB monomers form a triangle with the tip of the amino-terminal CC of one molecule trapped within the hydrophobic cavity of the carboxy-terminal domain of a second molecule. This arrangement suggests an analogous model for recruitment to RNAP. Indeed, the beta'-subunit CC located at the rim of the secondary channel has conserved hydrophobic residues at its tip. We show that substitutions of these residues and those in the GreB C-terminal domain cavity confer defects in GreB activity and binding to RNAP, and present a plausible model for the RNAP-GreB complex. PMID- 17917676 TI - Reduced in vitro functional activity of human NRAMP1 (SLC11A1) allele that predisposes to increased risk of pediatric tuberculosis disease. AB - Polymorphic variants within the human natural resistance-associated macrophage protein-1 (NRAMP1, also known as SLC11A1) gene have been shown to impact on susceptibility to tuberculosis in different human populations. In the mouse, Nramp1 is expressed at the macrophage phagosomal membrane and its activity can be assayed by the relative acquisition of mannose 6-phosphate receptor (M6PR) in Salmonella-containing vacuoles. Based on this M6PR recruitment assay, we have now developed an assay in primary human macrophages to test the function of human NRAMP1 gene variants. First, we established that M6PR acquisition was significantly higher (P = 0.002) in human U-937 monocytic cell lines transfected with NRAMP1 as compared to untransfected U-937 cells. Second, the M6PR assay was shown to be highly reproducible for NRAMP1 activity in monocyte-derived macrophages (MDM) from healthy volunteers. Finally, the assay was investigated in MDM from pediatric tuberculosis patients and significantly lower NRAMP1 activity was detected in MDM from individuals homozygous for the NRAMP1-274 high-risk allele (CC genotype) in comparison to heterozygous individuals (CT genotype; P=0.013). The present study describes both an assay for human NRAMP1 functional activity and concomitant evidence for reduced NRAMP1 function in the common genetic variant shown to be associated with tuberculosis susceptibility in pediatric patients. PMID- 17917677 TI - Circulating MCP-1 levels shows linkage to chemokine receptor gene cluster on chromosome 3: the NHLBI family heart study follow-up examination. AB - Atherogenesis is a chronic inflammatory process. Critical in the inflammation process is monocyte chemoattractant protein-1 (MCP-1). To locate genomic regions that affect circulating MCP-1 levels, a genome-wide linkage scan was conducted in a sample of whites and blacks. Phenotype and genetic marker data were available for 2501 white and 513 black participants in the National Heart Lung Blood Institute Family Heart Study follow-up examination. Heritability for MCP-1 was 0.37 in whites and 0.47 in blacks after adjusting for the effects of sex, age, age-sex interaction, smoking status, lifetime smoking exposure (pack-years) and field center. Significant linkage was observed for MCP-1 in a combined black and white sample on chromosome 3 (logarithm of the odds ratio (LOD)=3.5 at 78 cM, P=0.0001) and suggestive linkage was observed in whites on chromosome 5 (LOD=1.8 at 128 cM, P=0.002). Located under the linkage peak on chromosome 3 is the chemokine receptor gene cluster, including CCR2, the receptor for MCP-1. This study provides preliminary evidence linking genetic variation in a receptor to circulating levels of its ligand, as previously demonstrated for the low-density lipoprotein receptor. Further characterization of these chromosomal regions is needed to identify the functional mutations associated with circulating levels of MCP-1. PMID- 17917678 TI - ALS: astrocytes take center stage, but must they share the spotlight? PMID- 17917679 TI - Autophagy promotes necrosis in apoptosis-deficient cells in response to ER stress. PMID- 17917681 TI - The shadow of death on the MET tyrosine kinase receptor. AB - The MET tyrosine kinase receptor is a high-affinity receptor for hepatocyte growth factor/scatter factor (HGF/SF). HGF/SF-MET system is necessary for embryonic development, and aberrant MET signalling favours tumorigenesis and metastasis. MET is a prototype of tyrosine kinase receptor, which is able to counteract apoptosis through the initiation of a survival signal involving notably the PI3K-Akt pathway. Paradoxically, the MET receptor is also able to promote apoptosis when activated by HGF/SF or independently of ligand stimulation. The molecular mechanisms underlying this uncommon response have been recently investigated and revealed dual antiapoptotic or proapoptotic property of MET according to the cell type or stress conditions. Although the involvement of MET in the regulation of integrated biological responses mostly took into account its efficient antiapoptotic function, its proapoptotic responses could also be important for regulation of the survival/apoptosis balance and play a role during the development or tumour progression. PMID- 17917680 TI - Oxidative stress induces autophagic cell death independent of apoptosis in transformed and cancer cells. AB - Autophagy is a self-digestion process that degrades intracellular structures in response to stresses leading to cell survival. When autophagy is prolonged, this could lead to cell death. Generation of reactive oxygen species (ROS) through oxidative stress causes cell death. The role of autophagy in oxidative stress induced cell death is unknown. In this study, we report that two ROS-generating agents, hydrogen peroxide (H(2)O(2)) and 2-methoxyestradiol (2-ME), induced autophagy in the transformed cell line HEK293 and the cancer cell lines U87 and HeLa. Blocking this autophagy response using inhibitor 3-methyladenine or small interfering RNAs against autophagy genes, beclin-1, atg-5 and atg-7 inhibited H(2)O(2) or 2-ME-induced cell death. H(2)O(2) and 2-ME also induced apoptosis but blocking apoptosis using the caspase inhibitor zVAD-fmk (benzyloxycarbonyl-Val Ala-Asp fluoromethylketone) failed to inhibit autophagy and cell death suggesting that autophagy-induced cell death occurred independent of apoptosis. Blocking ROS production induced by H(2)O(2) or 2-ME through overexpression of manganese superoxide dismutase or using ROS scavenger 4,5-dihydroxy-1,3-benzene disulfonic acid-disodium salt decreased autophagy and cell death. Blocking autophagy did not affect H(2)O(2)- or 2-ME-induced ROS generation, suggesting that ROS generation occurs upstream of autophagy. In contrast, H(2)O(2) or 2-ME failed to significantly increase autophagy in mouse astrocytes. Taken together, ROS induced autophagic cell death in transformed and cancer cells but failed to induce autophagic cell death in non-transformed cells. PMID- 17917682 TI - Optic disc pit as evaluated with en-face optical coherence tomography: report of a case. PMID- 17917683 TI - Mean intraocular pressure and progression based on corneal thickness in patients with ocular hypertension. AB - PURPOSE: To determine the incidence of glaucomatous progression at mean intraocular pressure (IOP) levels in patients with ocular hypertension (OHT). METHODS: A retrospective, multicentre, cohort analysis of 230 OHT patients with 5 years of follow-up evaluated for risk factors associated with progressive optic disc and visual field loss to determine the incidence of glaucomatous progression. RESULTS: Forty percent of patients with IOPs > or = 24 mmHg, 18% of patients with IOPs of 21-23 mmHg, 11% of patients with IOPs with 18-20 mmHg, and 3% of patients with IOPs of < or = 17 mmHg progressed to glaucoma. The mean IOP was 19.8+/-2.4 mmHg in the stable group and 21.7+/-2.6 mmHg in the progressed group (P=0.0004). The highest average peak IOP was 23.4+/-4.0 mmHg in the stable group and 25.2+/-3.1 mmHg in the progressed group (P=0.006). Based on the pachymetry values for central corneal thickness, patients with thinner corneas more often progressed to glaucoma (P<0.0001). A multivariant regression analysis to determine risk factors for progression was positive primarily for higher peak IOPs, older age, male gender, argon laser trabeculoplasty, visual acuity > or = 20/50, and no topical medical therapy or beta-blocker therapy prior to the study. CONCLUSIONS: IOP reduction within the normal range over 5 years of follow-up reduces the chance of progression to primary open-angle glaucoma in OHT patients. PMID- 17917684 TI - Prenatal diagnosis of dacryocystocele. PMID- 17917686 TI - Subconjunctival injection of triamcinolone in the treatment of lid retraction of patients with thyroid eye disease: a case series. AB - PURPOSE: To report the results of subconjunctival injection of triamcinolone in the treatment of thyroid eye disease-related lid retraction. INTERVENTION: Patients with either unilateral or bilateral upper lid retraction, secondary to thyroid eye disease, diagnosed during the period of February 2004 to June 2005 were recruited. An injection of 0.5 ml of triamcinolone acetonide (40 mg/ml kenalog) with 0.1 ml of 2% lignocaine was injected into the subconjunctival region of the lid between the conjunctiva and Muller's muscle under topical anaesthesia on upper lid eversion. Pre- and post-procedure measurements included lid aperture, marginal reflex distance, the amount of lagophthalmos, and intraocular pressure measurements. Photographs were also obtained before the procedure and at subsequent visits. Follow-up was done at 2 weeks, 1, 3, 6 months and at 1 year. RESULTS: Three of the four patients had resolution of their upper lid retraction within 1 month of treatment, with one patient requiring a repeat triamcinolone injection. The patient who had fibrotic muscles did not respond to triamcinolone injections and required surgical correction. CONCLUSION: Upper lid subconjunctival triamcinolone appears to be an effective treatment option in reducing lid retraction in patients with recent onset of thyroid eye disease. PMID- 17917685 TI - Immediate vs delayed sequential cataract surgery: a comparative study. AB - PURPOSE: Immediate sequential cataract surgery (ISCS) is still a controversial procedure. We compared the clinical and patient-reported outcomes of ISCS vs the usual delayed sequential cataract surgery (DSCS). METHODS: Selected patients with bilateral cataracts were assigned nonrandomly and according to their preferences to either ISCS in one session (intervention group) or DSCS over two sessions with a 2-month interval between sessions (control group). Binocular visual acuity, binocular contrast sensitivity, stereopsis, and self-reported visual function (VF 14) were measured preoperatively and postoperatively for up to 4 months after the second-eye surgery. Mann-Whitney U-test was used to compare quantitative variables, while the chi(2) and Fisher's exact tests were used for qualitative variables. RESULTS: Of the 220 eligible patients, 74 (33.6%) chose ISCS, and 137 (62.3%) selected DSCS. The remaining patients (4.1%) were allocated randomly to either group. For 2 months, the outcome measures were significantly worse in the DSCS group than in the ISCS group. However, after 4 months (2 and 4 months after second-eye surgery in the DSCS and ISCS groups, respectively) the differences became insignificant except for VF-14 (P<0.05). The mean post-operative objective measures and their differences from baseline were not significantly different between the groups. Nevertheless, postoperatively, VF-14 improved more (P<0.05) and attained a higher value (P<0.05) in the ISCS group. CONCLUSIONS: In experienced hands, with stringent patient selection criteria and with a strict aseptic protocol, ISCS can safely provide a more rapid rehabilitation of VF than DSCS. PMID- 17917687 TI - Separation of the effects of pH and polymer concentration on the swelling pressure and elastic modulus of a pH-responsive hydrogel. AB - The elastic shear modulus G and swelling pressure omega are studied for a basic, pH-responsive hydrogel synthesized by crosslinking copolymerization of co monomers hydroxypropyl methacrylate and N,N-dimethylaminoethyl methacrylate with crosslinker tetraethylene glycol dimethacrylate. Under normal conditions of use as a "smart" material, hydrogel swelling ratio Q and pH vary simultaneously, but here G and omega values are presented as a function of pH with Q held constant and vice-versa. At fixed pH, G decreases with increase in Q in a power law dependence, as predicted by the Flory-Rehner model. However, at fixed Q, G increases with decrease in pH (i.e, increase in degree of ionization). The pH effect is more pronounced than the volume effect, thus the hydrogel stiffens as it swells in response to pH change. At high pH, omega values of the uncharged hydrogel obey the Flory-Rehner model, whereas explicit ionic contributions can be identified at lower pH values. PMID- 17917688 TI - Phytochemicals from Echinacea and Hypericum: A Direct Synthesis of Isoligularone. AB - Reaction of trienes with alpha,beta-unsaturated aldehydes produces bicyclic products via a tandem Diels-Alder/ene reaction. The adduct from tiglic aldehyde was converted into isoligularone by conversion to a furan followed by benzylic oxidation. PMID- 17917689 TI - Preliminary Study of OCD and Health Disparities at the U.S.-Mexico Border. AB - The widespread and devastating nature of obsessive compulsive disorder (OCD) sharply contrasts with the paucity of research involving Mexican Americans and Mexicans who suffer from this condition. This mixed-methods preliminary study was intended to provide initial data and to pilot the procedures for a larger investigation of the cultural identification, symptomatology, health concerns, coping mechanisms, and quality of life of Mexican Americans and Mexicans with OCD living in the U.S.-Mexico border region of El Paso, Texas and Ciudad Juarez, Mexico. For the sample of six participants, whose symptoms ranged from moderate to extreme, OCD was associated with marked impairment in quality of life, particularly in terms of social functioning, an area of central importance for many Mexican Americans and Mexicans. Areas of further study were identified, with the aim of developing culturally sensitive interventions to decrease health disparities involving OCD. PMID- 17917690 TI - Structure determination by restrained molecular dynamics using NMR pseudocontact shifts as experimentally determined constraints. AB - The structure of a DNA octamer d(TTGGCCAA)(2) complexed to chromomycin-A(3) and a single divalent cobalt ion has been solved by using the pseudocontact shifts due to the unpaired electrons on the cobalt. A protocol was developed and critically evaluated for using the pseudocontact shifts in structure determination. The pseudocontact shifts were input as experimental restraints in molecular dynamics simulations with or without NOE constraints. Both the magnitude and orientation of the susceptibility anisotropy tensor required for the shift calculations were determined during the simulations by iterative refinement. The pseudocontact shifts could be used to define the structure to a very high precision and accuracy compared with a corresponding NOE-determined structure. Convergence was obtained from different starting structures and tensors. A structure determination using both NOE's and pseudocontact shifts revealed a general agreement between the two data sets. However, some evidence for a discrepancy between NOE's and pseudocontact shifts was observed in the backbone and terminal base pairs of the DNA. Violations in shift or NOE restraints remaining in the final structures were examined and may be a reflection of motional averaging of the constraints and evidence for flexibility. This work demonstrates that pseudocontact shifts are a powerful tool for NMR structure determination. PMID- 17917692 TI - A Parametric k-Means Algorithm. AB - The k points that optimally represent a distribution (usually in terms of a squared error loss) are called the k principal points. This paper presents a computationally intensive method that automatically determines the principal points of a parametric distribution. Cluster means from the k-means algorithm are nonparametric estimators of principal points. A parametric k-means approach is introduced for estimating principal points by running the k-means algorithm on a very large simulated data set from a distribution whose parameters are estimated using maximum likelihood. Theoretical and simulation results are presented comparing the parametric k-means algorithm to the usual k-means algorithm and an example on determining sizes of gas masks is used to illustrate the parametric k means algorithm. PMID- 17917691 TI - Genetic markers and biomarkers for age-related macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of visual impairment and blindness in the USA. Although the treatment of AMD has evolved to include laser photocoagulation, photodynamic therapy, surgical macular translocation and antiangiogenesis agents, treatment options for advanced AMD are limited. Furthermore, the dry form of AMD, albeit less devastating than the wet form, has even fewer viable treatment options. This review summarizes the various biomarkers of AMD and analyzes whether or not they may one day be exploited to determine risks of disease onset, measure progression of disease or even assess the effects of treatment of AMD. Potential biomarkers are important to identify since some might be utilized to reflect the disease state of a particular patient and to individualize therapy. Although studies have yielded promising results for nutrient and inflammatory biomarkers, these results have been inconsistent. At present, the best available markers of AMD risk are single nucleotide polymorphisms (SNPs). SNPs in complement factor H (CFH) and PLEKHA1/ARMS2/HtrA1 capture a substantial fraction of AMD risk and permit the identification of individuals at high risk of developing AMD. PMID- 17917694 TI - Halting in Single Word Production: A Test of the Perceptual Loop Theory of Speech Monitoring. AB - The perceptual loop theory of speech monitoring (Levelt, 1983) claims that inner and overt speech are monitored by the comprehension system, which detects errors by comparing the comprehension of formulated utterances to originally intended utterances. To test the perceptual loop monitor, speakers named pictures and sometimes attempted to halt speech in response to auditory (Experiments 1 and 3) or visual (Experiments 2, 4, and 5) words that differed from the picture name. These stop-signal words were varied in terms of their semantic or phonological similarity to the intended word. The ability to halt word production was sensitive to phonological similarity and, in Experiment 5, to emotional valence, but not to semantic similarity. These results suggest that the perceptual loop detects errors by making comparisons at a level where phonological knowledge is represented. These data also imply that dialogue, back channeling, and other areas where speech production is affected by simultaneous comprehension may operate based on phonological comparisons. PMID- 17917695 TI - Elevated blood-lead levels in first nation people of Northern Ontario Canada: policy implications. AB - We evaluated the preliminary impact of the Canadian "non-toxic" shotshell policy, for the hunting of migratory game birds, by examining blood-lead levels of First Nations people living in sub-arctic Canada. If the use of lead shotshell was the major source of lead exposure as has been postulated and the ban on the use of lead shotshell for hunting migratory birds was immediately effective, we would expect that blood-lead levels would be typical of a geographic area remote from industrialization. Our findings present some concern in that approximately 18% of the 196 First Nations people examined had blood-lead levels > or =100 microg/L. PMID- 17917696 TI - Creatine supplementation reduces plasma levels of pro-inflammatory cytokines and PGE2 after a half-ironman competition. AB - OBJECTIVE: The effect of creatine supplementation upon plasma levels of pro inflammatory cytokines: Interleukin (IL) 1 beta and IL-6, Tumor Necrosis Factor alpha (TNFalpha), and Interferon alpha (INF alpha) and Prostaglandin E(2) (PGE(2)) after a half-ironman competition were investigated. METHODS: Eleven triathletes, each with at least three years experience of participation in this sport were randomly divided between the control and experimental groups. During 5 days prior to competition, the control group (n = 6) was supplemented with carbohydrate (20 g x d(-1)) whereas the experimental group (n = 5) received creatine (20 g x d(-1)) in a double-blind trial. Blood samples were collected 48 h before and 24 and 48 h after competition and were used for the measurement of cytokines and PGE(2). RESULTS: Forty-eight hours prior to competition there was no difference between groups in the plasma concentrations (pg x ml(-1), mean +/- SEM) of IL-6 (7.08 +/- 0.63), TNFalpha (76.50 +/- 5.60), INF alpha (18.32 +/- 1.20), IL-1 beta (23.42 +/- 5.52), and PGE(2) (39.71 +/- 3.8). Twenty-four and 48 h after competition plasma levels of TNFalpha, INF alpha, IL-1 beta and PGE(2) were significantly increased (P < 0.05) in both groups. However, the increases in these were markedly reduced following creatine supplementation. An increase in plasma IL-6 was observed only after 24 h and, in this case, there was no difference between the two groups. CONCLUSION: Creatine supplementation before a long distance triathlon competition may reduce the inflammatory response induced by this form of strenuous of exercise. PMID- 17917697 TI - Suggestive evidence for chromosomal localization of non-coding RNA from imprinted LIT1. AB - The non-coding RNA LIT1/KCNQ1OT1, itself the product of an imprinted gene, is involved in cis-limited silencing within an imprinted cluster on human chromosome 11p15.5. Although the locus serves as an imprinting center, the mechanism of transcriptional regulation is not clear. To help understand the function of the LIT1 non-coding RNA, we used fluorescence in situ hybridization (FISH) to examine the sub-cellular localization of LIT1 RNA molecules. LIT1 RNA signals were observed in most of the interphase human lymphoblast and fibroblast cells. The RNA also appeared to accumulate on neighboring regions of chromatin containing the SLC22A18/IMPT1 and CDKN1C/p57KIP2 genes, as shown by high-resolution fiber RNA FISH and modified RNA TRAP (tagging and recovery of associated proteins) methods. These results suggest that LIT1 RNA stably localizes to a specific chromatin region and plays an important role in the transcriptional silencing of the imprinting domain. PMID- 17917698 TI - Different effects of galactose and mannose on cell proliferation and intracellular soluble sugar levels in Vigna angularis suspension cultures. AB - Plant cells utilize various sugars as carbon sources for growth, respiration and biosynthesis of cellular components. Suspension-cultured cells of azuki bean (Vigna angularis) proliferated actively in liquid growth medium containing 1% (w/v) sucrose, glucose, fructose, arabinose or xylose, but did not proliferate in medium containing galactose or mannose. These two latter sugars thus appeared distinct from other sugars used as growth substrates. Galactose strongly inhibited cell growth even in the presence of sucrose but mannose did not, suggesting a substantial difference in their effects on cell metabolism. Analysis of intracellular soluble-sugar fractions revealed that galactose, but not mannose, caused a conspicuous decrease in the cellular level of sucrose with no apparent effects on the levels of glucose or fructose. Such a galactose-specific decrease in sucrose levels also occurred in cells that had been cultured together with glucose in place of sucrose, suggesting that galactose inhibits the biosynthesis, rather than uptake, of sucrose in the cells. By contrast, mannose seemed to be metabolically inert in the presence of sucrose. From these results, we conclude that sucrose metabolism is important for the heterotrophic growth of cells in plant suspension-cultures. PMID- 17917699 TI - "Recreational" drug abuse associated with failure to mount a proper antibody response after a generalised orthopoxvirus infection. AB - Infections with orthopoxviruses usually lead to cross-protection among all species of the family. This has been a prerequisite for successful eradication of smallpox. Here we report the rare case of a 17-year-old male, who survived a generalised cowpox virus infection of unusual severity but surprisingly did not show a proper seroconversion. Only a very weak antibody production was observed in early and late serum samples, which initially appeared to be cowpox virus specific in immunofluorescence. No neutralising antibodies were detected and in Western blotting antibody specificity was restricted to the orthopoxvirus H3L protein only. The patient had been hospitalised for alcohol and cannabis intoxication 2 months prior to the orthopoxvirus infection and high levels of cannabinoids have been found repeatedly in the urine and upon one occasion also benzodiazepines. As these substances are known to interfere with antibody production and no immunodeficiencies were detected, drug-induced immunosuppression can be suspected as the most likely cause. Therefore a possible link between "soft" drug use and sufficient immunosuppression to warrant alterations in vaccine policies using live virus vaccines like smallpox vaccine should be further studied. PMID- 17917700 TI - A cold-active salmon goose-type lysozyme with high heat tolerance. AB - The Atlantic salmon (Salmo salar) goose-type lysozyme gene was isolated and revealed alternative splicing within exon 2 affecting the signal peptide-encoding region. The lysozyme was produced in Escherichia coli, and the recombinant enzyme showed a high specific lytic activity that was stimulated by low or moderate concentrations of mono- or divalent cations. Relative lytic activities of 70 and 100% were measured at 4 degrees C and 22 degrees C, respectively, and there was no detectable activity at 60 degrees C. However, 30% activity was retained after heating the enzyme for 3 h at 90 degrees C. This unique combination of thermal properties was surprising since the salmon goose-type lysozyme contains no cysteines for protein structure stabilization through disulphide bond formation. The results point to a rapid reversal of inactivation, probably due to instant protein refolding. PMID- 17917701 TI - DNA fragmentation in leukocytes following repeated low dose sarin exposure in guinea pigs. AB - The objective of this study was to determine levels of DNA fragmentation in blood leukocytes and parietal cortex from guinea pigs following repeated low-level exposure to the chemical warfare nerve agent (CWNA) sarin. Guinea pigs were injected (s.c.) once a day for 10 days with saline, or 0.1, 0.2, or 0.4 LD50 (50% mean lethal dose) sarin dissolved in sterile physiological saline. Blood and parietal cortex was collected after injection at 0, 3, and 17 days recovery and evaluated for DNA fragmentation using single-cell gel electrophoresis (Comet assay). Cells were imaged using comet analysis software and three parameters of DNA fragmentation measured: tail length, percent DNA in the tail, and tail moment arm. Repeated low-dose exposure to sarin produced a dose-dependent response in leukocytes at 0 and 3 days post-exposure. There was a significant increase in all measures of DNA fragmentation at 0.2 and 0.4 LD50, but not at 0.1 LD50. There was no significant increase in DNA fragmentation in any of the groups at 17 days post exposure. Sarin did not produce a systematic dose-dependent response in parietal cortex at any of the time points. However, significant increases in DNA fragmentation at 0.1 and 0.4 LD50 were observed at 0 and 3 days post-exposure. All measures of DNA fragmentation in both leukocytes and neurons returned to control levels by 17 days post-exposure, indicating a small and non-persistent increase in DNA fragmentation following repeated low-level exposure to sarin. PMID- 17917703 TI - Construction of bifunctional fusion proteins consisting of duck BAFF and EGFP. AB - We constructed fusion proteins consisting of fluorescence-enhanced green fluorescent protein (EGFP) and soluble domain of duck B-cell-activating factor of the TNF family (dsBAFF). The soluble EGFP/dsBAFF was efficiently expressed in Escherichia coli BL 21 (DE3) and was purified in milligram amounts using metal chellate affinity chromatography. The fusion protein exhibited similar fluorescence spectra with free EGFP and promoted the survival of duck bursal B cells in vitro as well as dsBAFF. EGFP/dsBAFF has shown specific binding to duck BAFF receptors positive-cells and the stained cells could be analyzed with flow cytometry. Thus, the fusion protein represents a readily obtainable source of biologically active dsBAFF that may prove useful in further studies on duck BAFF and its receptors. PMID- 17917704 TI - Microbial degradation of chlorinated benzenes. AB - Chlorinated benzenes are important industrial intermediates and solvents. Their widespread use has resulted in broad distribution of these compounds in the environment. Chlorobenzenes (CBs) are subject to both aerobic and anaerobic metabolism. Under aerobic conditions, CBs with four or less chlorine groups are susceptible to oxidation by aerobic bacteria, including bacteria (Burkholderia, Pseudomonas, etc.) that grow on such compounds as the sole source of carbon and energy. Sound evidence for the mineralization of CBs has been provided based on stoichiometric release of chloride or mineralization of (14)C-labeled CBs to (14)CO(2). The degradative attack of CBs by these strains is initiated with dioxygenases eventually yielding chlorocatechols as intermediates in a pathway leading to CO(2) and chloride. Higher CBs are readily reductively dehalogenated to lower chlorinated benzenes in anaerobic environments. Halorespiring bacteria from the genus Dehalococcoides are implicated in this conversion. Lower chlorinated benzenes are less readily converted, and mono-chlorinated benzene is recalcitrant to biotransformation under anaerobic conditions. PMID- 17917702 TI - The role of bystin in embryo implantation and in ribosomal biogenesis. AB - Human bystin was identified as a cytoplasmic protein directly binding to trophinin, a cell adhesion molecule potentially involved in human embryo implantation. Although the trophinin gene is unique to mammals, the bystin gene (BYSL) is conserved across eukaryotes. Recent studies show that bystin plays a key role during the transition from silent trophectoderm to an active trophoblast upon trophinin-mediated cell adhesion. Bystin gene knockout and knockdown experiments demonstrate that bystin is essential for embryonic stem cell survival and trophectoderm development in the mouse. Furthermore, biochemical analysis of bystin in human cancer cells and mouse embryos indicates a function in ribosomal biogenesis, specifically in processing of 18S RNA in the 40S subunit. Strong evidence that BYSL is a target of c-MYC is consistent with a role for bystin in rapid protein synthesis, which is required for actively growing cells. PMID- 17917705 TI - Degradation of mixtures of phenolic compounds by Arthrobacter chlorophenolicus A6. AB - In this study the chlorophenol-degrading actinobacterium, Arthrobacter chlorophenolicus A6, was tested for its ability to grow on mixtures of phenolic compounds. During the experiments depletion of the compounds was monitored, as were cell growth and activity. Activity assays were based on bioluminescence output from a luciferase-tagged strain. When the cells were grown on a mixture of 4-chlorophenol, 4-nitrophenol and phenol, 4-chlorophenol degradation apparently was delayed until 4-nitrophenol was almost completely depleted. Phenol was degraded more slowly than the other compounds and not until 4-nitrophenol and 4 chlorophenol were depleted, despite this being the least toxic compound of the three. A similar order of degradation was observed in non-sterile soil slurries inoculated with A. chlorophenolicus. The kinetics of degradation of the substituted phenols suggest that the preferential order of their depletion could be due to their respective pKa values and that the dissociated phenolate ions are the substrates. A mutant strain (T99), with a disrupted hydroxyquinol dioxygenase gene in the previously described 4-chlorophenol degradation gene cluster, was also studied for its ability to grow on the different phenols. The mutant strain was able to grow on phenol, but not on either of the substituted phenols, suggesting a different catabolic pathway for the degradation of phenol by this microorganism. PMID- 17917706 TI - Effect of stable weak magnetic field on Cr(VI) bio-removal in anaerobic SBR system. AB - To study the impact of stable weak magnetic field on the Cr(VI) removal efficiency of predominated strains in ASBR system, the choice of the optimum magnetic density and its effect should be considered chiefly. At different magnetic densities, the growth and propagation rates of predominated strains in solid or liquid mediums and their capabilities of removing Cr(VI) were compared. The results showed that the optimum magnetic density was 6.0 mT. To meet the state first-class standard of effluent discharge, it took 2-5 h more in the plant wastewater treatment than in the synthetic wastewater treatment, but the presence of magnetic field made the reaction time up to par to decrease 1 and 2-3 h, respectively, compared with that of the control. The magnetized magnetic powder could improve the sludge sedimentation capability, turbidity of outflow water and efficiency of bio-system. PMID- 17917707 TI - Lack of association between soluble CD40L and risk in a large cohort of patients with acute coronary syndrome in OPUS TIMI-16. AB - BACKGROUND: Previous studies have suggested that elevated soluble CD40 ligand (sCD40L) levels predict adverse cardiovascular outcomes in patients with acute coronary syndromes (ACS). Recently, questions have been raised regarding the influence of pre-analytical and analytical conditions on measurement of sCD40L, and additional studies have had conflicting findings regarding the prognostic value of this marker. METHODS AND RESULTS: We measured levels of sCD40L in citrated plasma using an analytically validated automated immunoassay (Roche Diagnostics) in a large cohort of ACS patients (n = 2403) from the placebo arm of the OPUS TIMI-16 trial. No association was observed between elevated sCD40L levels and risk of death or myocardial infarction (MI) (Quartile 1, 8.0%; Quartile 2, 11.7%; Quartile 3, 8.2%; Quartile 4, 6.8%; P = 0.54) or risk of death, MI or heart failure at 10 months (Quartile 1, 9.9%; Quartile 2, 14.2%; Quartile 3, 10.9%; Quartile 4, 8.3%; P = 0.55). A comparison of plasma vs. serum measurements of sCD40L was performed on samples from a nested case-control analysis (n = 42) from within this cohort. Median sCD40L levels did not differ between cases and controls using plasma (0.23 ng/ml vs. 0.27 ng/ml, respectively; P = 0.82) or serum samples (0.64 vs. 0.77, respectively; P = 0.85). Serum samples consistently yielded elevated sCD40L measurements compared to plasma samples (median value 0.72 ng/ml vs. 0.25 ng/ml, respectively, P < 0.001). CONCLUSIONS: The absence of an association between sCD40L and cardiovascular outcomes in a large cohort of patients with ACS raises concern regarding the reproducibility of clinical results with this novel biomarker. Despite a plausibly important role in the pathobiology of atherothrombosis, pre-analytic sources of variability may limit the practical clinical application of sCD40L. PMID- 17917708 TI - Effect of high dose statin therapy on platelet function; statins reduce aspirin resistant platelet aggregation in patients with coronary heart disease. AB - BACKGROUND: Current evidence supports the preventive role of statins on platelet aggregation in patients with coronary heart disease. AIM: Our aim was to determine the effects of aggressive statin therapy on platelet function in patients with coronary heart disease. MATERIAL AND METHODS: A total of 178 consecutive patients (37-68 years old, 35.9% women) with stable coronary artery disease (CAD) was enrolled in the study. Platelet function assays were realized by the Platelet Function Analyzer (PFA)-100 with collagen and epinephrine (Col/Epi) and collagen and ADP (Col/ADP) cartridges. Aspirin resistance was defined as having a closure time (CT) of <186 s with Col/Epi cartridges despite regular aspirin therapy. A statin therapy protocol applied to the patients with aspirin resistance for 3 months. RESULTS: We determined that 20 (11.2%) of patients had aspirin resistance by the PFA-100. Mean closure time measured with the Col/ADP cartridges was 83 +/- 18 s (53-162 s). Of the patients 12 were not on a statin therapy and eight were taking 10 mg daily atorvastatin. After 3 months of 40 mg daily atorvastatin therapy 13 subjects with aspirin resistance became aspirin sensitive by PFA-100 (P < 0.0001). There was also a significant decrease in total and LDL cholesterol levels and an increase in HDL cholesterol at the third month of statin therapy (P < 0.0001 for all). CONCLUSION: Statin therapy reduced the in vitro aspirin resistance in 65% of the patients after a therapy of 3 months. Further studies are needed to elucidate the mechanism of statins' effects on platelet reactivity. PMID- 17917710 TI - [31st Fall Meeting of the German Society of Cardiology and the 18th Annual Meeting of the Cardiac Pacemaker and Arrhythmia Working Group, 2007. Abstract database]. PMID- 17917709 TI - In vivo performance of an oral MR matrix tablet formulation in the beagle dog in the fed and fasted state: assessment of mechanical weakness. AB - PURPOSE: To evaluate the behaviour of an oral matrix modified release formulation in the canine gastrointestinal tract, and establish if a mechanical weakness previously observed in clinical studies would have been identified in the dog model. MATERIALS AND METHODS: In vitro release profiles were obtained for two modified release matrix tablets containing UK-294,315, designed to release over either 6 (formulation A) or 18 (formulation B) hours. Tablets were labelled with (153)samarium and in vivo pharmacoscintigraphy studies were performed in four beagle dogs in the fasted state for both formulations, and following ingestion of an FDA high fat meal for formulation B. RESULTS: The matrix tablet formulations displayed significantly different in vitro release profiles (F (2) < 50), with time to 80% release for formulation A and B of 406 and 987 min respectively. Complete in vivo disintegration occurred at 339 +/- 181 and 229 +/- 171 for formulation A and B respectively in the fasted state, and at 207 +/- 154 min for formulation B in the fed state, in disagreement with in vitro release. CONCLUSION: The fed/fasted dog model would have predicted a lack of physical robustness in the matrix tablet formulation B, however it would not have predicted the clear fed/fasted effects on performance observed previously in man. PMID- 17917712 TI - Comment on: Gale EAM (2007) To boldly go -- or to go too boldly? The accelerator hypothesis revisited. Diabetologia 50:1571-1575 -- a reply to the editor. PMID- 17917711 TI - Variation in the FTO gene locus is associated with cerebrocortical insulin resistance in humans. PMID- 17917713 TI - Continuous terlipressin infusion in patients with persistent septic shock and cirrhosis of the liver. PMID- 17917714 TI - Improving standards of oral hygiene in intensive care. PMID- 17917716 TI - Therapeutic effects of the putative P2X3/P2X2/3 antagonist A-317491 on cyclophosphamide-induced cystitis in rats. AB - It is suggested that ATP and purinergic P2X receptors are involved in overactive bladder. In this study, we investigated the effect of the recently developed P2X3 and P2X2/3 receptor antagonist A-317491 on cyclophosphamide (CYP)-induced cystitis to determine whether a P2X receptor antagonist could be beneficial for the treatment of bladder overactivity induced by CYP. Female Sprague-Dawley (SD) rats were given 150 mg/kg CYP (i.p.). When the micturition activity was observed for 24 h in a conscious and unrestrained condition, CYP-treated rats exhibited increased urinary frequency. Two days after CYP injection, cystometry was performed in conscious rats, in which the bladder was continuously infused with saline (5 ml/h). In CYP-treated rats, non-voiding contractions were interposed between micturitions, suggestive of hyper-reflexia. Intravenous administration of A-317491 (20 or 50 mg/kg) or pyridoxal phosphate-6-azo (benzene-2,4-disulfonic acid) tetrasodium (PPADS; a nonselective purinergic receptor antagonist, 10 mg/kg) prolonged the interval of voiding contraction and reduced the non-voiding contractions. On the other hand, oxybutynin (1 mg/kg), a muscarinic receptor antagonist, did not affect the frequency of non-voiding or voiding contractions in CYP-treated rats. A-317491 at the higher dose decreased the amplitude of voiding contractions, but increased the micturition volume. The residual urine in the bladder increased after treatment with CYP; A-317491 and PPADS reduced this, whereas oxybutynin had no effect. These data suggest that A-317491 is effective at improving the signs of CYP-induced cystitis and that the P2X3 or P2X2/3 receptor pathway is involved in bladder overactivity observed during CYP-induced cystitis. PMID- 17917717 TI - Mechanisms of carvedilol-induced [Ca2+] i rises and death in human hepatoma cells. AB - The effect of the cardiovascular drug carvedilol on cytosolic free Ca2+ concentrations ([Ca2+]i) and viability has not been explored in human hepatoma cells. This study examined whether carvedilol altered [Ca2+]i and caused cell death in HA59T cells. [Ca2+]i and cell viability were measured using the fluorescent dyes fura-2 and WST-1, respectively. Carvedilol at concentrations >or=1 microM increased [Ca2+]i in a concentration-dependent manner with an EC50 value of 20 microM. The Ca2+ signal was reduced partly by removing extracellular Ca2+. Carvedilol induced Mn2+ quench of fura-2 fluorescence, implicating Ca2+ influx. The Ca2+ influx was sensitive to La3+, econazole, nifedipine, and SKF96365. In Ca2+-free medium, after pretreatment with 1 muM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor), carvedilol-induced [Ca2+]i rises were abolished; and conversely, carvedilol pretreatment inhibited a major part of thapsigargin-induced [Ca2+]i rises. Inhibition of phospholipase C with 2 microM U73122 did not change carvedilol-induced [Ca2+]i rises. At concentrations between 1 and 50 microM, carvedilol killed cells in a concentration-dependent manner. The cytotoxic effect of 1 microM (but not 30 microM) carvedilol was fully reversed by prechelating cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N' tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Apoptosis was induced by 30 (but not 1) microM carvedilol. Collectively, in HA59T hepatoma cells, carvedilol induced [Ca2+]i rises by causing Ca2+ release from the endoplasmic reticulum in a phospholipase-C-independent manner and Ca2+ influx via store-operated Ca2+ channels. Carvedilol-caused cytotoxicity was mediated by Ca2+ and apoptosis in a concentration-dependent manner. PMID- 17917718 TI - Modulation of human risky decision making by flunitrazepam. AB - RATIONALE: GABA-modulating drugs produce disinhibitory effects that increase the probability of risk-taking behavior. Previous reports suggest that the misuse of the benzodiazepine flunitrazepam is associated with several forms of harmful risky behavior, including theft, violence, and intoxication-related auto accidents. OBJECTIVES: The present study examined the dose-response relationships between acute flunitrazepam administration and human decision making under conditions of risk. The analyses also examined flunitrazepam-mediated changes in decision-making processes using a computational modeling approach, the expectancy valence model (EVM). MATERIALS AND METHODS: Using a laboratory measure of risky decision making designed to address acute drug effects, 12 adults were administered placebo, 0.5, 1.0, and 2.0 mg/70 kg flunitrazepam in a within subject, repeated measures counterbalanced design. Flunitrazepam was compounded and doses were administered in an 8-oz liquid solution. Primary data analyses examined subjective effects, response rates, distribution of choices between the risky and nonrisky option, and personality correlates related to peak drug effects. Individual-subject data were submitted to a computational modeling analysis (EVM) that provided parameter estimates corresponding to components of valence; updating expectancies about alternatives (learning/memory); and consistency between choices and expected outcomes (sensitivity to learned outcomes). RESULTS: Flunitrazepam produced dose-related changes in subjective effects and response rates, and increased selection of the risky response option. High doses significantly changed decision-making processes related to the learning/memory and consistency parameters. CONCLUSIONS: At sufficiently high doses, flunitrazepam can engender increases in risky decision making. Globally, these changes appear similar to previous effects we have observed after acute administration of alcohol and alprazolam. As suggested by the EVM outcomes, the mechanisms underlying the changes in risky decision making are more similar to alprazolam than alcohol. PMID- 17917720 TI - Application of enriched stable isotopes as tracers in biological systems: a critical review. AB - The application of enriched stable isotopes of minerals and trace elements as tracers in biological systems is a rapidly growing research field that benefits from the many new developments in inorganic mass spectrometric instrumentation, primarily within inductively coupled plasma mass spectrometry (ICP-MS) instrumentation, such as reaction/collision cell ICP-MS and multicollector ICP-MS with improved isotope ratio measurement and interference removal capabilities. Adaptation and refinement of radioisotope tracer experiment methodologies for enriched stable isotope experiments, and the development of new methodologies coupled with more advanced compartmental and mathematical models for the distribution of elements in living organisms has enabled a broader use of enriched stable isotope experiments in the biological sciences. This review discusses the current and future uses of enriched stable isotope experiments in biological systems. PMID- 17917722 TI - Interaction behaviour of a PDMS-calixarene system and polar analytes characterised by microcalorimetry and spectroscopic methods. AB - Spectroscopic techniques and microcalorimetry were applied to investigate a polymer-(polydimethylsiloxane; PDMS) calixarene system during interaction with propylamine and n-propanol as analyte molecules. This was done to understand the sensitivity and selectivity of this system. By these means the interesting binding site of the calixarene selector was identified and dependencies on specific properties of the polymer and the functional groups were determined. Reflectometric interference spectroscopy (RIfS) was used to characterize the kinetics whereas isothermal titration calorimetry (ITC) yielded thermodynamic data. Infrared (IR) and (1)H NMR spectroscopy allowed identification of the sensing process as an interaction between the selective group of the PDMS calixarene system and the amino group of propylamine, and measurement of the effects on hydrogen bonds. The combination of the different spectroscopic methods and the microcalorimetric measurements broadened the understanding of this system, regarded as a model system. Thus, future tailoring of functional groups designed for improved and more selective analyte detection is possible. PMID- 17917719 TI - Dose-related effects of the acetylcholinesterase inhibitor tacrine on cocaine and food self-administration in rats. AB - RATIONALE: Acetylcholine (ACh) is involved in brain reward and learning functions and contributes to opiate- and psychostimulant-motivated behaviors. Tacrine is a centrally acting, reversible cholinesterase inhibitor that also inhibits monoamine oxidase (MAO) and blocks reuptake of dopamine (DA) and serotonin. OBJECTIVES: To determine the effects of pretreatment with tacrine on self administration of cocaine and nondrug reinforcers. MATERIALS AND METHODS: Male Wistar rats were trained to self-administer cocaine under a fixed-ratio-5 (FR-5) schedule during 2-h multiple-component sessions in which 0.1, 0.2, and 0.4 mg/kg per injection of cocaine were each available for 40 min. Other animals self administered 45 mg food pellets under FR-30 or 20% Ensure (liquid food) under FR 5 in amounts of 30, 60, or 120 microl. Vehicle or tacrine was administered as single intravenous doses 20 min before self-administration of cocaine, food pellets, or liquid food. RESULTS: Although pretreatment with 0.032 mg/kg of tacrine increased self-administration of food pellets, pretreatment with higher doses of tacrine attenuated self-administration of cocaine, food pellets, or liquid food. Tacrine's ED50 value for attenuating self-administration of 0.1 mg/kg per injection of cocaine was more than sixfold lower than values for attenuating liquid food- or food pellet-reinforced behavior. However, ED50 values for attenuating self-administration of higher doses of cocaine were similar to those observed for 30 or 60 microl of liquid food. CONCLUSIONS: Tacrine can selectively attenuate self-administration of low-dose cocaine, but its effects on higher doses of cocaine are similar to its ability to decrease self administration of nondrug reinforcers. PMID- 17917724 TI - Dextranomer/hyaluronic acid copolymer (Deflux) implants mimicking distal ureteral calculi on CT. AB - Periureteral or subtrigonal injection of dextranomer/hyaluronic acid (Dx/HA) copolymer (Deflux, Q-Med, Uppsala, Sweden) is an increasingly common endoscopic treatment for vesicoureteral reflux. We report a confusing radiographic finding of bilateral calcified Dx/HA injections initially thought to represent bilateral distal ureteral stones in a boy who presented with intermittent periumbilical pain. Urologists, radiologists, and emergency room physicians should be aware of the potential for calcification of ureteral implants of Dx/HA, and of the potentially confusing radiographic images that may result. PMID- 17917723 TI - Intra-arterial thrombolysis using rt-PA in patients with acute stroke due to vessel occlusion of anterior and/or posterior cerebral circulation. AB - INTRODUCTION: The aim of our study was to evaluate the safety and efficacy of intra-arterial (IA) thrombolysis using recombinant tissue plasminogen activator (rt-PA) in patients with acute stroke due to occlusion in the anterior or posterior circulation. METHODS: We retrospectively analyzed the clinical and radiological data of 88 consecutive patients with acute ischemic stroke who underwent emergency cerebral angiography for the purpose of subsequent IA thrombolysis. The neurological deficit on admission and discharge was graded using the National Institutes of Health Stroke Scale (NIHSS) score. Baseline computer tomography (CT) scans were examined for any signs indicative of cerebral ischemia. The angiographic findings were classified according to the Thrombolysis in Myocardial Infarction (TIMI) score for myocardial infarction. Follow-up CT scans were examined for hemorrhagic complication. RESULTS: Of the 88 patients who underwent IA thrombolysis, 63 presented with complete or partial arterial occlusion in the suspected perfusion area. In these 63 patients, the median NIHSS score dropped from 15 points on admission to 10 points at discharge. The recanalization rate was 52.6% for partial and complete reperfusion. In-hospital mortality was 20.6% (9.1% for carotid, 44.4% for basilar territory occlusion). Intracerebral bleeding (ICB) occurred in 38.6% of the patients with occlusion in the anterior circulation, resulting in these patients presenting a worse clinical outcome than those without ICB. Only minor extracranial bleedings occurred in 20.6% of patients. Patients with ICB had a significantly higher frequency of ischemic signs on the baseline CT scan. CONCLUSION: Occlusion of a cerebral artery is present in about 75% of the patients eligible for thrombolytic therapy. Intra-arterial thrombolysis using rt-PA in patients with acute ischemic stroke can achieve re-vascularization, although ICB remains the major risk factor affecting its efficacy. PMID- 17917725 TI - Reactive blue 19 decolouration by laccase immobilized on silica beads. AB - Laccase (31.5 U of activity/g or 4.39 mug of protein/m(2)) from Trametes versicolor was immobilized on controlled-porosity-carrier silica beads and evaluated for the decolouration of Reactive blue 19, an anthraquinone dye. Although there was an initial, rapid adsorption of the dye to the packed bed in a recirculating reactor, about 97.5% of Reactive blue 19 removal was due to enzymatic degradation. The free enzyme lost 52% of its activity in 48 h. However, the activity of the immobilized laccase was unchanged after 4 months of storage in phosphate buffer under ambient conditions followed by three successive decolourations over 120 h. Treating the laccase immobilized beads with ethanolamine reduced dye adsorption by 40%. PMID- 17917726 TI - Enzymatic synthesis and characterization of arbutin glucosides using glucansucrase from Leuconostoc mesenteroides B-1299CB. AB - Two arbutin glucosides were synthesized via the acceptor reaction of a glucansucrase from Leuconostoc mesenteroides B-1299CB with arbutin and sucrose. The glucosides were purified by Bio-gel P-2 column chromatography and high performance liquid chromatography, and the structures were elucidated as 4 hydroxyphenyl beta-isomaltoside (arbutin-G1), 4-hydroxyphenyl beta isomaltotrioside (arbutin-G2), according to the results of (1)H, (13)C, heteronuclear single-quantum coherence, (1)H-(1)H COSY, and heteronuclear multiple-bond correlation analyses. Arbutin glucoside (4-hydroxyphenyl beta isomaltoside) exhibited slower effects on 1,1-diphenyl-2-picrylhydrazyl radical scavenging and similar effects on tyrosinase inhibition, and increased inhibitory effect on matrix metalloproteinase-1 production induced by UVB than arbutin. PMID- 17917728 TI - Absent immunoglobulins in HIV-related Burkitt lymphoma/leukaemia. PMID- 17917730 TI - Diagnostic accuracy of the apparent diffusion coefficient in differentiating benign from malignant uterine endometrial cavity lesions: initial results. AB - Our purpose is to evaluate the diagnostic accuracy of apparent diffusion coefficient (ADC) measurement in differentiating malignant from benign uterine endometrial cavity lesions. We retrospectively evaluated 25 uterine endometrial cavity lesions in 25 female patients: endometrial carcinoma (n = 11), carcinosarcoma (n = 2), submucosal leiomyoma (n = 8), and endometrial polyp (n = 4). Diffusion-weighted images were performed at 1.5 T with b factors of 0 1,000/mm(2). The region of interest was defined within the tumor on T2-weighted EPI image and then manually copied to an ADC map. Thereby, the ADC value was obtained. We compared ADC values between malignant and benign lesions using Student's t-test. The mean and standard deviation of ADC values (x10(-3) mm(2)/s) were as follows: endometrial carcinoma, 0.98+/-0.21; carcinosarcoma, 0.97+/-0.02; submucosal leiomyoma, 1.37+/-0.28; and endometrial polyp, 1.58+/-0.45. The ADC values differed significantly between malignant (0.98+/-0.19) and benign lesions (1.44+/-0.34) (P < 0.01). We defined malignant tumors as cases with an ADC value less than 1.15 x 10(-3) mm(2)/s for obtaining the highest accuracy. Sensitivity, specificity, and accuracy were 84.6%, 100%, and 92%, respectively. ADC measurement can provide useful information in differentiating malignant from benign uterine endometrial cavity lesions. PMID- 17917729 TI - Guarana (Paullinia cupana var. sorbilis), an anciently consumed stimulant from the Amazon rain forest: the seeded-fruit transcriptome. AB - Guarana (Paullinia cupana var. sorbilis) is a plant native to the central Amazon basin. Roasted seed extracts have been used as medicinal beverages since pre Colombian times, due to their reputation as stimulants, aphrodisiacs, tonics, as well as protectors of the gastrointestinal tract. Guarana plants are commercially cultivated exclusively in Brazil to supply the national carbonated soft-drink industry and natural product stores around the world. In this report, we describe and discuss the annotation of 15,387 ESTs from guarana seeded-fruits, highlighting sequences from the flavonoid and purine alkaloid pathways, and those related to biotic stress avoidance. This is the largest set of sequences registered for the Sapindaceae family. PMID- 17917731 TI - When to perform CTA in patients suspected of PE? AB - Venous thromboembolic disease (VTE) is a common disorder which may be associated with high morbidity or mortality when left untreated. Specific VTE diagnosis is mandatory, as treatment is associated with significant side effects. Therefore, timely diagnostic tests are necessary to establish the presence or absence of VTE. Computed tomographic pulmonary angiography (CTPA) has reached a high accuracy in the evaluation of pulmonary embolism (PE). Unfortunately, the continuous decrease of the prevalence of PE in the most recent studies can lead to cost-efficacy imbalance and overuse of ionizing radiation when CTPA is used as a single test. Therefore, no single non-invasive test is suitable for all patients and diagnostic strategies based on sequential non-invasive tests are likely to identify patients in whom anticoagulation can be withheld safely and limit the number of patients requiring more invasive or more expensive tests. The cost effectiveness of clinical stratification and D-dimer test has been demonstrated as it reduces the requirement for invasive tests. In this paper, the current role of CTPA in the diagnosis of PE will be reviewed. PMID- 17917734 TI - Effectiveness of colorectal laparoscopic surgery on patients at high anesthetic risk: an intervention cohort study. AB - AIMS: The aim of the study was to assess the effectiveness of laparoscopic colorectal surgery in patients at high preoperative anesthetic risk because of associated pathologies. MATERIALS AND METHODS: From January 2003 until January 2005, 116 patients were systematically assigned at a ratio of 1:1 to one of two groups: laparoscopy surgery (n=59, of which 31 were American Society of Anesthesiologists score [ASA] I-II [L1] and 28 ASA III-IV [L2]) or open surgery (n=57, of which 30 were ASA I-II [O1] and 27 ASA III-IV [O2]). Data on patient demographics and clinical and anesthetic variables were collected prospectively. Informed consent was obtained from the patients, and approval was obtained from the designated review board of the institution involved. RESULTS: The number of minor anesthetic complications during surgery was higher in L2 patients. No differences were observed in blood gas parameters studied during surgery (pCO(2), pH, and pO(2)/FiO(2)). Transfusion rates in the laparoscopy group at greater anesthetic risk (L2) were lower than those of the high-risk conventional surgery group (O2; 21.4 vs 63%, P<0.02). Duration of stay in the surgical recovery room and the inpatient ward were also shorter in the L2 group than in the O2 group (8.7+/-4.5 vs 12.2+/-6 days, P=0.02). There was no difference in perioperative clinical variables between laparoscopy groups (L1, L2). CONCLUSION: Postoperative recovery of ASA III-IV patients is better after laparoscopic surgery for colorectal cancer, at the expense of a higher rate of minor anesthetic occurrences during surgery. PMID- 17917732 TI - In vivo imaging of transplanted hepatocytes with a 1.5-T clinical MRI system- initial experience in mice. AB - The feasibility of in vitro mature mouse hepatocyte labeling with a novel iron oxide particle was assessed and the ability of 1.5-T magnetic resonance imaging (MRI) to track labeled mouse hepatocytes in syngenic recipient livers following intraportal cell transplantation was tested. Mouse hepatocytes were incubated with anionic iron oxide nanoparticles at various iron concentrations. Cell viability was assessed and iron oxide particle uptake quantified. Labeled hepatocytes were intraportally injected into 20 mice, while unlabeled hepatocytes were injected into two mice. Liver T2 values, spleen-to-muscle relative signal intensity (RI( spleen/muscle )), and liver-to-muscle relative signal intensity (RI( liver/muscle )) on gradient-echo T2-weighted imaging after injection of either labeled or unlabeled hepatocytes were compared with an ANOVA test followed by Fisher's a posteriori PLSD test. Livers, spleens and lungs were collected for histological analysis. Iron oxide particle uptake was saturable with a maximum iron content of 20 pg per cell and without viability alteration after 3 days of culture. Following labeled-cell transplantation, recipient livers showed well defined nodular foci of low signal intensity on MRI--consistent with clusters of labeled hepatocytes on pathological analysis--combined with a significant decrease in both liver T2 values and liver-to-muscle RI( liver/muscle ) (P = 0.01) with minimal T2 values demonstrated 8 days after transplantation. Conventional MRI can demonstrate the presence of transplanted iron-labeled mature hepatocytes in mouse liver. PMID- 17917735 TI - Modified single-portal type II SLAP repair. AB - We present a simplified, cost-effective method for repairing a type II SLAP lesion that requires only one working portal in the rotator interval. The rotator cuff tendon or muscle is not violated when using this portal. The biceps root can be firmly reattached anteriorly and posteriorly using one double-loaded absorbable bone anchor with a suture eyelet. By retrieving the anterior limbs of the anchor percutaneously using a spinal needle and PDS suture, tangling of the anchor suture or premature knot formation are avoided during shuttling and knot tying. PMID- 17917733 TI - Antipyretic treatment of noninfectious fever in children with severe traumatic brain injury. AB - OBJECTIVE: The purpose of this study was to describe the treatment of noninfectious fever in children with severe traumatic brain injury (TBI). MATERIALS AND METHODS: We conducted a retrospective study to compare type of and response to antipyretic treatment strategies in children less than or equal to 17 years and Glasgow Coma Scale (GCS) score less than 9. RESULTS: The average admission GCS score was 4. Forty children (35 boys, 5 girls), age 7.8 +/- 5.2 years, had noninfectious fever. Seventy percent (28 of 40) received acetaminophen only, and 30% (12 of 40) received acetaminophen plus either ibuprofen or physical cooling. Time to next febrile episode was longer in patients receiving combination therapy than those receiving monotherapy (p = 0.03). Fever refractory to treatment dose or strategy occurred in more than 40% of the patients. CONCLUSIONS: Early combination antipyretic therapy may be needed to effectively maintain normothermia in children with severe TBI. PMID- 17917736 TI - The role of genetic susceptibility in head and neck squamous cell carcinoma. AB - Our research is an additional genetic study to uncover the molecular mechanisms involved in head and neck squamous cell carcinoma (HNSCC) pathogenesis by studying loss of heterozygosity (LOH) and microsatellite instability (MSI) in both premalignant and malignant patients and to highlight the genotype of HNSCC in Upper Egypt. Patients with HNSCC from various parts of the world may have unique genotypes and this is the first genetic study of HNSCC in Sohag 500 KM to the south of Cairo. We performed a prospective study of 41 patients with precancerous and 79 patients with cancerous laryngeal, esophageal, nasopharyngeal, nasal and oral lesions, and 50 controls (The control patients were cases admitted for ear surgery or simple nasal surgery, from whom we took biopsy from mucosal lining of nasopharynx). The present study included 170 individuals who were admitted to the Ear, Nose and Throat department, Sohag University Hospital, Sohag, in Egypt in the period between April 2001 and March 2003. Samples which were taken by punch biopsy were frozen and stored at -80 degrees C and were subjected to histopathological examination. We investigated LOH and MSI by using six microsatellite markers located at chromosomes 3, 5, 9, and 17. The markers used were D3S1286, D9S171, D9S753, D17S654, D17S695, and CFS1 R. LOH was in all premalignant and malignant lesions at 5q33.3-q34 and 13% of Controls. LOH at 17p21 was absent in all premalignant lesions and was found in 53% of malignant lesions and 12.4% of Controls. In premalignant lesions, LOH was at 3pter-3p24.2 (73% of cases), at 9p21 (46%), at 9q21.1-22.3 (37%), and at 17p13 (37%). These percents increased in malignant lesions to 87, 80, 67, and 63%, respectively. They were 14, 19.4, 17, and 19% in controls. Examination of LOH could improve diagnosis, adds additional confidence, in HNSCC by DNA extraction from suspicious lesions in high-risk groups (smokers and alcoholics) and LOH at 3p/9p seems to be of particular value for early detection and definition of progression risk. If there are high percent of LOH at these chromosomes, active intervention should be done (chemoprevention and regular follow up head and neck examination for very early detection and management). PMID- 17917737 TI - Study of age dependence of epiphyseal ossification of the hand skeleton. AB - Radiological analysis of the hand skeleton is a key pillar of forensic age diagnostics in living subjects undergoing criminal proceedings. The present study investigated whether ossification stage classification of selected epiphyses of the hand could provide added value to hand radiograph analysis. Hand radiographs from 265 male and 164 female subjects aged 10-18years old who had been X-rayed due to traumatological indications were therefore assessed. Epiphyseal ossification of selected elements of the hand skeleton (ulna, radius and third metacarpal, basal phalanx, mesophalanx and telephalanx) was graded based on the criteria of the five-stage classification system of Schmeling et al. (Int J Legal Med, 118:5-8, 2004) for clavicular epiphyseal cartilage. Stage 5 (absence of the epiphyseal scar) does not occur in the radius of men before the age of 18. Stage 5 can therefore be regarded as a potential parameter for valid determination of a minimum age of 18 years for forensic age estimation in criminal proceedings. PMID- 17917738 TI - Intravitreal bevacizumab with or without triamcinolone for refractory diabetic macular edema; a placebo-controlled, randomized clinical trial. AB - PURPOSE: To evaluate the effect of three intravitreal injections of bevacizumab (IVB) alone or combined with triamcinolone (IVT) in the first injection for treatment of refractory diabetic macular edema (DME). METHODS: In this prospective, placebo-controlled, randomized clinical trial, 115 eyes of 101 patients with refractory DME were included. Subjects were randomly assigned to one of the three study arms: 1) three injections of IVB (1.25 mg/0.05 ml) at 6 week intervals, 2) combined IVB and IVT (1.25 mg/0.05 ml and 2 mg/0.05 ml respectively) followed by two injections of IVB at 6-week intervals, and 3) sham injection (control group). The primary outcome measure was change in central macular thickness (CMT). Secondary outcome measures were change in best-corrected logMAR visual acuity (BCVA ) and incidence of potential adverse events. RESULTS: Central macular thickness was reduced significantly in both the IVB and IVB/IVT groups. At week 24, CMT change compared to the baseline was -95.7 microm (95% CI, -172.2 to -19.26) in the IVB group, -92.1 microm (95% CI, -154.4 to -29.7) in the IVB/IVT group, and 34.9 microm (95% CI, 7.9 to 61.9) in the control group. There was a significant difference between the IVB and control groups (P = 0.012) and between the IVB/IVT and control groups (P = 0.022). Improvement of BCVA was initiated at weeks 6 and 12 in the IVB/IVT and IVB groups respectively. In terms of BCVA change compared to the baseline at 24 weeks, the differences between the IVB and control groups (P = 0.01) and also between the IVB/IVT and control groups (P = 0.006) were significant. No significant differences were detected in the changes of CMT and BCVA between the IVB and IVB/IVT groups (P = 0.99). Anterior chamber reaction was noticed in eight (19.5%) and seven (18.9%) eyes respectively in the IVB and IVB/IVT groups the day after injection, and it resolved with no sequel. Elevation of IOP occurred in three eyes (8.1%) in the IVB/IVT group. CONCLUSION: Three consecutive intravitreal injections of bevacizumab had a beneficial effect on refractory DME in terms of CMT reduction and BCVA improvement. Addition of triamcinolone in the first injection seemed to induce earlier visual improvement; however, it did not show any significant additive effect later during follow-up. PMID- 17917740 TI - Structural equation analysis of the causal relationship between health and perceived indoor environment. AB - OBJECTIVES: To explore the temporal relationship and reversed effects between health and perception of the indoor environment using structural equation models. METHODS: The study was a two-phase prospective questionnaire study with a cross lagged design. Altogether 1,740 adults participated on both occasions. RESULTS: The perceived indoor environment had only weak effects on health at follow-up. However, the results strongly indicated a reversed effect that health problems may lead to increased complaints about the indoor environment. CONCLUSIONS: Structural equation models are powerful analytical tools for disentangling the effects of a specific variable on another in high dimensional data with complex patterns of associations. The analyses confirmed the results of our previous logistic regression analysis about the strong reversed effect. Hence, it is probable that a reversed effect between health and complaints about the indoor environment exists. PMID- 17917739 TI - Clinical features of early and late stage polypoidal choroidal vasculopathy characterized by lesion size and disease duration. AB - BACKGROUND: The pathogenesis of polypoidal choroidal vasculopathy (PCV), and even its clinical features, are controversial. Previous histopathological studies have identified different features; either dilated choroidal vessels or intra-Bruch's neovascularization. These differences might be partly attributable to the influence of the disease stage. We therefore evaluated the clinical features of early and late stage PCV. METHODS: The medical records of 110 eyes of 97 PCV patients were retrospectively reviewed. The time between the subjective onset of visual abnormality and examination at our clinic and the greatest linear dimension of the total lesion at the first examination were investigated. The period of disturbed vision and lesion size data were placed in ascending order to determine the first quartile point. Eyes with both values at or below the first quartile point were classified as 'small-short' (early stage). Eyes with both values equal to at least the third quartile point were classified as 'large-long' (late stage). Fundus photography, indocyanine green and fluorescein angiography, visual acuity, and clinical course were compared. RESULTS: Twelve eyes from 12 patients were small-short cases (period of disturbed vision of 1 month or less, lesion size 2.0 disc diameters or less). Eleven eyes from ten patients were large long cases (period of disturbed vision 36 months or more, lesion size at least 5.0 disc diameters). The large-long eyes were characterized by occult choroidal neovascular membrane or scar tissue secondary to exudative age-related macular degeneration. Noticeable in the small-short eyes were atrophic changes in the retinal pigment epithelium, choroidal vessel hyperpermeability and pulsation. The visual prognosis and clinical course were different between the groups. CONCLUSIONS: The difference of clinical features between the groups might reflect different disease stages, although not all of the features observed in the small short group appeared to represent the early stages of those recorded in the large long group. Thus, the variation in histopathologic features among previous reports might be partly attributable to differences in disease stage. PMID- 17917741 TI - Stage IV early gastric cancer: two cases with microsatellite instability. AB - BACKGROUND: We experienced two rare cases of stage IV early gastric carcinoma with extensive lymph node metastasis (pT1N3) and microsatellite instability caused by methylation of the MLH1 gene. MATERIALS AND METHODS: The clinicopathological features of patients with stage IV EGC were examined. Microsatellite instability (MSI) was analyzed using five microsatellite markers and MLH1 gene promoter methylation patterns were determined by methylation specific polymerase chain reaction (PCR) from paraffin-embedded tissue samples. Loss of hMLH1 was identified by immunohistochemical study. RESULTS: In both cases, the carcinomas were located in the antrum of the stomach, confined within the submucosa (SM3) and accompanied by endolymphatic tumor emboli. Immunohistochemical analysis for hMLH1 showed negative nuclear staining in the carcinoma cells. The tumor demonstrated methylation of the MLH1 gene and MSI-H because they manifested instability in three and two of the five markers tested. One patient is alive without any clinical, radiological, or pathological evidence of recurrence or metastasis at 37 months, while the other patient died after a cerebrovascular accident 5 months after surgery. CONCLUSION: Our rare cases support that MSI caused by MLH1 promoter methylation and the loss of hMLH1 protein play an important role in stage IV EGC development. PMID- 17917742 TI - Increased polycomb-group oncogene Bmi-1 expression correlates with poor prognosis in hepatocellular carcinoma. AB - PURPOSE: Recent studies have identified polycomb-group gene Bmi-1 as oncogene in the generation of mouse pre-cell lymphomas, and overexpression of Bmi-1 has been found in several human tumor with the disease progress and poor prognosis of the cancer patients. METHODS: In present study, we investigated Bmi-1 expression and its prognostic significance in hepatocellular carcinoma (HCC) by performing immunohistochemical analysis, using a total of 137 HCC clinical tissue samples. RESULTS: High Bmi-1 expression (Bmi-1 2+ or 3+) was shown in 29.9% cases. The positive immuno-staining of Bmi-1 was not only in well/moderately-differentiated tumor cells, but also in surrounding noncancerous or cirrhotic liver tissue. Bmi 1 expression level did not correlate with any clinicopathological parameters. However, survival analysis showed that the high-Bmi-1 group had a significantly shorter overall survival time than the low-Bmi-1 group (P=0.047). Multivariate analysis after 24 months revealed that Bmi-1 expression was a significant and independent prognostic parameter (P=0.002) for HCC patients. CONCLUSIONS: Our study indicated that Bmi-1 could be a candidate biomarker for long-term survival in HCC. PMID- 17917743 TI - Identification and characterization of secreted and pathogenesis-related proteins in Ustilago maydis. AB - Interactions between plants and fungal pathogens require a complex interplay at the plant-fungus interface. Extracellular effector proteins are thought to play a crucial role in establishing a successful infection. To identify pathogenesis related proteins in Ustilago maydis we combined the isolation of secreted proteins using a signal sequence trap approach with bioinformatic analyses and the subsequent characterization of knock-out mutants. We identified 29 secreted proteins including hydrophobins and proteins with a repetitive structure similar to the repellent protein Rep1. Hum3, a protein containing both, a hydrophobin domain and a repetitive Rep1-like region, is shown to be processed during passage through the secretory pathway. While single knock-outs of hydrophobin or repellent-like genes did not affect pathogenicity, we found a strong effect of a double knock-out of hum3 and the repetitive rsp1. Yeast-like growth, mating, aerial hyphae formation and surface hydrophobicity were unaffected in this double mutant. However, pathogenic development in planta stops early after penetration leading to a complete loss of pathogenicity. This indicates that Hum3 and Rsp1 are pathogenicity proteins that share an essential function in early stages of the infection. Our results demonstrate that focusing on secreted proteins is a promising way to discover novel pathogenicity proteins that might be broadly applied to a variety of fungal pathogens. PMID- 17917745 TI - Control of cultivation processes for recombinant protein production: a review. AB - The current state-of-the-art in control of cultivation processes for recombinant protein production is examined including the quantitative knowledge that can be activated for this purpose and the measurement techniques that can be employed for control at industrial manufacturing sites. PMID- 17917744 TI - Identification of InuR, a new Zn(II)2Cys6 transcriptional activator involved in the regulation of inulinolytic genes in Aspergillus niger. AB - The expression of inulinolytic genes in Aspergillus niger is co-regulated and induced by inulin and sucrose. We have identified a positive acting transcription factor InuR, which is required for the induced expression of inulinolytic genes. InuR is a member of the fungal specific class of transcription factors of the Zn(II)2Cys6 type. Involvement of InuR in inulin and sucrose metabolism was suspected because of the clustering of inuR gene with sucB, which encodes an intracellular invertase with transfructosylation activity and a putative sugar transporter encoding gene (An15g00310). Deletion of the inuR gene resulted in a strain displaying a severe reduction in growth on inulin and sucrose medium. Northern analysis revealed that expression of inulinolytic and sucrolytic genes, e.g., inuE, inuA, sucA, as well as the putative sugar transporter gene (An15g00310) is dependent on InuR. Genome-wide expression analysis revealed, three additional putative sugar transporters encoding genes (An15g04060, An15g03940 and An17g01710), which were strongly induced by sucrose in an InuR dependent way. In silico analysis of the promoter sequences of strongly InuR regulated genes suggests that InuR might bind as dimer to two CGG triplets, which are separated by eight nucleotides. PMID- 17917746 TI - Quality of end-of-life treatment for cancer patients in general wards and the palliative care unit at a regional cancer center in Japan: a retrospective chart review. AB - GOALS: In Japan, most cancer patients die in the hospital. The aim of this study was to assess the quality of end-of-life treatment for dying cancer patients in general wards and palliative care unit (PCU). MATERIALS AND METHODS: A retrospective chart review study was conducted. The following data on cancer patients who died in general wards (N = 104) and PCU (N = 201) at a regional cancer center were collected: do-not-resuscitate (DNR) decisions, treatments in the last 48 h of life, and aggressiveness of cancer care for dying patients. MAIN RESULTS: DNR orders were documented for most patients (94% in general wards, 98% in PCU, p = 0.067) and families usually consented (97%, 97%, p = 0.307). Comparison of general wards with PCU showed that, in the last 48 h of life, significantly more patients in general wards received life-sustaining treatment (resuscitation, 3.8%, 0%, p = 0.001; mechanical ventilation, 4.8%, 0%, p = 0.004), large volume hydration (>1,000 ml/day, 67%, 10%, p < 0.001) with continuous administration (83%, 5%, p = 0.002) and fewer palliative care drugs (strong opioids, 68%, 92%, p < 0.001; corticosteroids, 49%, 70%, p < 0.001; nonsteroidal anti-inflammatory drugs, 34%, 85%, p < 0.001). Regarding aggressiveness of cancer care, patients received a new chemotherapy regimen within 30 days of death (3.0%), chemotherapy within 14 days of death (4.3%), and intensive care unit admission in the last month of life (3.3%). CONCLUSION: We found that families, not patients, consented to DNR, and life-sustaining treatments were appropriately withheld; however, patients on general wards received excessive hydration, and the use of palliative care drugs could be improved. Application of our findings can be used to improve clinical care in general wards. PMID- 17917748 TI - Homology modeling and examination of the effect of the D92E mutation on the H5N1 nonstructural protein NS1 effector domain. AB - Virulent H5N1 strains of influenza virus often harbor a D92E point mutation in the nonstructural protein NS1. This crucial mutation has been correlated with increased virulence and/or cytokine resistance, but the structural implications of such a change are still unclear. Furthermore, NS1 protein could also be a potential target for the development of novel antiviral agents against H5N1 strains. Therefore, a reasonable 3D model of H5N1 NS1 is important for the understanding of the molecular basis of increased virulence and the design of novel antiviral agents. Based on the crystal structure of a non-H5N1 NS1 protein, a model of H5N1 NS1 was developed by homology modeling, molecular mechanics and molecular dynamics simulations. It was found that the D92E mutation could result in weakened interactions of the carboxylate side chain with other phosphorylated residues, thereby activating phosphorylation of NS1. PMID- 17917747 TI - Preoperative sterilization and disinfection of drill guide templates. AB - The aim of the in vitro study was to evaluate the decontamination potential of common antiseptic solutions for heat-sensitive implantological drill guide templates. One hundred implantologists were evaluated on the basis of a questionnaire for their measures of disinfection. On the basis of these results, 80% alcohol, Octenidine 0.1%, and Chlorhexidine 0.12% were tested in an in vitro model for their decontamination efficacy for heat-sensitive plastic material infected with Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Enterobacter cloacae, Escherichia coli, and Candida albicans. The microorganisms were selected on the basis of results of environmental testing of dental laboratories. The results of the questionnaire revealed that Chlorhexidine was used by 30%, 80% alcohol by 23%, and Octenidine by 7% of the dentists. Using the in vitro model, with the exception of S. aureus, Chlorhexidine was not able to completely eliminate the microorganisms after 15 min of application. In contrast, the treatment with Octenidine revealed no further growth of the tested microorganisms after that time. The 80% alcohol was more efficient. No growth of microorganisms could be detected in any of the tests after 5 min of incubation. On the basis of our results and due to the fact that suitable installations for sterilization were hardly used by the dental practitioners, the disinfection of templates should be preferentially performed with 80% alcohol or Octenidine using an incubation time of 15 min with ultrasonication. PMID- 17917749 TI - Density functional theory characterisation of 4-hydroxyazobenzene. AB - We report the structural properties, infrared (IR) and Raman spectra, dipole moment, polarisability, hardness and chemical potential of the trans and cis configurations of 4-hydroxyazobenzene calculated using the B3LYP functionals. All calculations were performed with the following basis sets: 6-31G, 6-31++G, 6 31G(d,p), 6-31++G(d,p), 6-31G(2d,2p), 6-31++G(2d,2p) and 6-311++G(2d,2p). We observed that 6-31++G(d,p) gives similar results to 6-311++G(2d,2p). Consequently, SVWN and PW91 methods were also used in association with 6 31++G(d,p) to test the influence of the different models of exchange and correlation functionals. A planar structure was obtained for all the optimised trans configuration structures. In both isomers, the presence of the hydroxyl group leads to an asymmetry in certain structural parameters. From these results, two IR or Raman active frequencies can be used to easily distinguish trans and cis configurations. The trans configuration was found to be more stable than the cis configuration by 67 +/- 2 kJ mol(-1) at 0 K. The difference of the dipole moment between trans and cis for 4-hydroxyazobenzene was found to be lower than for trans and cis azobenzene. PMID- 17917750 TI - Studies of the interaction of tetramethylcucurbit[6]uril and 5,5'-dimethyl-2,2' bipyridyl hydrochloride. AB - The interaction between tetramethylcucurbit[6]uril (host) and 5,5'-dimethyl-2,2' bipyridyl hydrochloride (guest) was studied by 1H NMR, X-ray crystallography, electronic absorption spectroscopy, fluorescence emission spectra and quantum chemistry calculations. This experimental-computational study that indicated the host can orientationally encapsulate the guest with a moderate association constant value. Computation qualitatively explained the split UV-visible absorption peak of the inclusion complex. PMID- 17917751 TI - Candidate genes for the progression of malignant gliomas identified by microarray analysis. AB - Malignant astrocytomas of World Health Organization (WHO) grade III or IV have a reduced median survival time, and possible pathways have been described for the progression of anaplastic astrocytomas and glioblastomas, but the molecular basis of malignant astrocytoma progression is still poorly understood. Microarray analysis provides the chance to accelerate studies by comparison of the expression of thousands of genes in these tumours and consequently identify targeting genes. We compared the transcriptional profile of 4,608 genes in tumours of 15 patients including 6 anaplastic astrocytomas (WHO grade III) and 9 glioblastomas (WHO grade IV) using microarray analysis. The microarray data were corroborated by real-time reverse transcription-polymerase chain reaction analysis of two selected genes. We identified 166 gene alterations with a fold change of 2 and higher whose mRNA levels differed (absolute value of the t statistic of 1.96) between the two malignant glioma groups. Further analyses confirmed same transcription directions for Olig2 and IL-13Ralpha2 in anaplastic astrocytomas as compared to glioblastomas. Microarray analyses with a close binary question reveal numerous interesting candidate genes, which need further histochemical testing after selection for confirmation. IL-13Ralpha2 and Olig2 have been identified and confirmed to be interesting candidate genes whose differential expression likely plays a role in malignant progression of astrocytomas. PMID- 17917752 TI - Dynamics of visuo-spatial remembering: a study of information structuring in memory. AB - We studied the process by which learning a pattern of motor activity reaches a steady-state characterized by a reduction in fluctuations. The stimuli consisted of eight visually presented dots that appeared sequentially. In a 20-trial learning phase, participants reproduced the positions of the eight dots after each presentation. Next, they reproduced the pattern 40 times without renewed presentation. In one condition, spatial distances between the dots were proportional to the intervals between their appearances; in the other they were not proportional. We analyzed how the reproduction stabilized at the configuration and dot levels. In proportional as well as non-proportional conditions, stabilization occurs at different time scales for the configuration and dot levels. The stabilization rate differed between proportional and non proportional conditions. These results are discussed in the framework of dynamical systems. PMID- 17917753 TI - The role of the episodic buffer in working memory for language processing. AB - A body of work has accumulated to show that the cognitive process of binding information from different mnemonic and sensory sources as well as in different linguistic modalities can be fractionated from general executive functions in working memory both functionally and neurally. This process has been defined in terms of the episodic buffer (Baddeley in Trends Cogn Sci 4(11):417-423, 2000). This paper considers behavioural, neuropsychological and neuroimaging data that elucidate the role of the episodic buffer in language processing. We argue that the episodic buffer seems to be truly multimodal in function and that while formation of unitary multidimensional representations in the episodic buffer seems to engage posterior neural networks, maintenance of such representations is supported by frontal networks. Although, the episodic buffer is not necessarily supported by executive processes and seems to be supported by different neural networks, it may operate in tandem with the central executive during effortful language processing. There is also evidence to suggest engagement of the phonological loop during buffer processing. The hippocampus seems to play a role in formation but not maintenance of representations in the episodic buffer of working memory. PMID- 17917755 TI - Automated live cell screening system based on a 24-well-microplate with integrated micro fluidics. AB - In research, pharmacologic drug-screening and medical diagnostics, the trend towards the utilization of functional assays using living cells is persisting. Research groups working with living cells are confronted with the problem, that common endpoint measurement methods are not able to map dynamic changes. With consideration of time as a further dimension, the dynamic and networked molecular processes of cells in culture can be monitored. These processes can be investigated by measuring several extracellular parameters. This paper describes a high-content system that provides real-time monitoring data of cell parameters (metabolic and morphological alterations), e.g., upon treatment with drug compounds. Accessible are acidification rates, the oxygen consumption and changes in adhesion forces within 24 cell cultures in parallel. Addressing the rising interest in biomedical and pharmacological high-content screening assays, a concept has been developed, which integrates multi-parametric sensor readout, automated imaging and probe handling into a single embedded platform. A life maintenance system keeps important environmental parameters (gas, humidity, sterility, temperature) constant. PMID- 17917754 TI - Computer simulation for the optimization of patient positioning in spinal deformity instrumentation surgery. AB - Studies have shown that scoliosis curves correct when patients are positioned on the operating table prior to instrumentation. However, biomechanical aspects of positioning have not been widely studied. The objective of this study was to simulate patient positioning during instrumentation surgery and test various adjustment parameters of the trunk and recommend optimal patient positioning prior to, and during spine surgery based on the results of finite element simulations. A scoliotic patient was simulated using a finite element model and six different positioning parameters were modified while ten geometric measures were recorded. Statistical analysis determined which model parameter had a significant effect on the geometric measures. Geometric measures were individually and simultaneously optimized, while corresponding model parameters were documented. Every model parameter had a significant effect on at least five of the geometric measures. When optimizing a single measure, others would often deteriorate. Simultaneous optimization resulted in improved overall correction of the patient's geometry by 75% however ideal correction was not possible for every measure. Finite element simulations of various positioning parameters enabled the optimization of ten geometric measures. Positioning is an important surgical step that should be exploited to achieve maximum correction. PMID- 17917756 TI - Modeling of skeletal muscle: the influence of tendon and aponeuroses compliance on the force-length relationship. AB - The aim of this study was to investigate the influence of changing elastic properties of tendon and aponeuroses on force production and muscle geometry. A three-dimensional, structural, continuum mechanics model of the cat medial gastrocnemius was used for this purpose. Increasing compliance in tendon and aponeuroses caused a decrease in the peak isometric force and a shift of the force-length relationship to the right of the length axis (i.e. toward greater muscle lengths). This result can be explained with the stability condition of the force-length relationship which produced a history dependence of force production that is conceptually in agreement with experimental observations. PMID- 17917757 TI - Evolution of receptors for growth hormone and somatolactin in fish and land vertebrates: lessons from the lungfish and sturgeon orthologues. AB - Two cognate hormones, growth hormone (GH) and somatolactin (SL), control several important physiological processes in vertebrates. Knowledge about GH and its receptor (GHR) has accumulated over the last decades. However, much less is known about SL and its receptor (SLR). SL is found only in fish (including lungfish), suggesting that it was present in the common ancestor of vertebrates, but was lost secondarily in the lineage leading to land vertebrates after the lungfish branched off. SLR was suggested to be a duplicated copy of GHR acquired only in teleosts via the fish-specific genome duplication (FSGD). This scenario (i.e., the existence of SL but not SLR in the vertebrate ancestors) is intriguing but contested. In this study, we first evaluated the plausibility of this scenario through synteny analyses and found that the loci for GHR and SLR are located in syntenic genomic positions, whereas the loci for GH and SL are not. Next, we cloned GHRs of lungfish and sturgeon, which possess SL but did not undergo the FSGD (i.e., they should not possess SLR). Their phylogenetic positions in the GHR/SLR gene tree further support the fish-specific scenario for the GHR-SLR duplication. Interestingly, their sequences share greater similarity with teleost SLRs and reptilian/amphibian GHRs than with the GHRs of mammals, birds, and teleosts. On the basis of these results, we discuss the validity of the nomenclature of the teleost-specific copy of GHR as SLR and an ancestral receptor(s) for SL before the evolution of SLR during the FSGD. PMID- 17917758 TI - Acute toxicity of waterborne Cd, Cu, Pb, Ni, and Zn to first-instar Chironomus riparius larvae. AB - The acute toxicities of waterborne Cd, Cu, Pb, Ni, and Zn were determined in the first-instar larvae (generally considered to be the most sensitive) of Chironomus riparius, under standardized conditions. Toxicity tests were conducted in soft water (hardness, 8 mg/L as CaCO(3) equivalents) in the absence of food and were limited to 24 h to avoid control mortality associated with food deprivation. For each metal, a logarithmic range of concentrations was tested between 0 and 25 mg/L. First-instar C. riparius are most sensitive to Pb, with a 24-h LC50 of 0.61 mg/L (CI of 0.26-1.15 mg/L), and to Cu, with a 24-h LC50 of 2.09 mg/L (CI of 1.57 2.96 mg/L). The LC50 for Cd was 9.38 mg/L, while the LC50s for Zn and Ni were both higher than the highest tested concentration of 25 mg metal/L. Our results found that even first-instar chironomid larvae are well protected by both the current CCME Canadian water quality guidelines for the protection of aquatic life and the USEPA Water Quality Criteria, as LC50s were at least 25 times higher than the guideline concentrations. PMID- 17917759 TI - Acute toxicity of aqueous copper, cadmium, and zinc to the mayfly Rhithrogena hageni. AB - Heptageniid mayfly nymphs have been suggested as sensitive indicators of metal contamination in streams based on biomonitoring studies, experimentation in situ, and experimentation in microcosm. Laboratory tests were conducted to evaluate the sensitivity of Rhithrogena hageni, a heptageniid mayfly, to waterborne copper, cadmium, and zinc. Tests were conducted with soft water (hardness = 40-50 mg/L) at about 12 degrees C. Toxicity endpoints were survival and moulting (%/day). Median 96 hr lethal concentrations were 0.137, 10.5, and 50.5 mg/L for copper, cadmium and zinc, respectively. The average daily moulting rate of survivors significantly decreased after exposure to these metals in solution. PMID- 17917760 TI - Acute and chronic effects of perfluorobutane sulfonate (PFBS) on the mallard and northern bobwhite quail. AB - Perfluorobutane sulfonate (PFBS) can be a final degradation product of perfluorobutane sulfonyl fluoride (PBSF)-based chemicals. Surfactants based on this chemistry are potential replacements for perfluorooctane sulfonate (PFOS) related products and have many potential applications in industrial and commercial processes and applications. To evaluate the potential hazard that PFBS may pose to avian species, acute dietary studies with juvenile mallards and northern bobwhite quail, as well as a quail dietary chronic study of reproduction were conducted. In the acute studies, 10-day-old mallards and quail were exposed to nominal dietary concentrations of 1,000, 1,780, 3,160, 5,620 or 10,000 mg PFBS/kg feed, wet weight (ww) for 5 days and the birds were then fed an untreated diet and observed for up to 17 days. No treatment-related mortalities were observed in the study up to 10,000 mg PFBS/kg, ww feed. Body weight gains of quail exposed to 5620 or 10,000 mg PFBS/kg feed were statistically less than that of unexposed controls. Weight gain of mallards exposed to 10,000 mg PFBS/kg feed was statistically less than that of controls. There were no statistically significant effects on feed consumption of either species. In the acute studies, no observed adverse effect concentration (NOAEC) for mallards and quail were 5620 and 3160 mg PFBS/kg, ww feed, respectively. In a reproduction study, adult quail were exposed to nominal dietary concentrations of 100, 300, or 900 mg PFBS/kg, ww feed for up to 21 weeks. There were no treatment-related mortalities or effects on body weight, weight gain, feed consumption, histopathology measures, or reproductive parameters evaluated in the study when compared to the control group. Concentrations of PFBS in blood serum, liver, and eggs were dose-dependent but were less than the administered dose, indicating biodiminution. Based on the results from the quail reproduction study, the dietary NOAEC was 900 mg PFBS/kg, ww feed (equivalent to an ADI of 87.8 mg PFBS/kg bw/d). PMID- 17917761 TI - Dioxin-like toxicity in the Saginaw River Watershed: polychlorinated dibenzo-p dioxins, dibenzofurans, and biphenyls in sediments and floodplain soils from the Saginaw and Shiawassee Rivers and Saginaw Bay, Michigan, USA. AB - Sediment and floodplain soils in the Saginaw River Watershed, Michigan, USA, have been demonstrated to be contaminated with a variety of organic compounds, including polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs). Existing data indicate that, at some locations, the contamination exceeds human health risk-based regulatory levels and ecological risk-based screening levels. In this study, concentrations of PCBs including non-ortho coplanar congeners, PCDDs, and PCDFs were measured in more than 120 sediment and floodplain soil samples collected from the Shiawassee River (a tributary of the Saginaw River), the Saginaw River, and Saginaw Bay, to determine the sources and magnitude of contamination, and to elucidate the contributions from individual contaminant groups to the overall 2,3,7,8 tetrachlorodibezo-p-dioxin equivalents (TEQs). Sediment and soil extracts were also analyzed for total dioxin-like activity by means of the H4IIE-luc cell bioassay. Elevated concentrations of PCBs (>1000 ng/g, dry weight) were found in surface sediment and floodplain soil samples collected from the Shiawassee River near Chase Lake, from Middleground Island in the Saginaw River, and from subsurface sediments in Saginaw Bay. Concentrations of PCDDs and PCDFs in sediment and floodplain soils from the Saginaw River and Saginaw Bay were 2 to 3 orders of magnitude higher than concentrations in the samples from the Shiawassee River. The highest PCDD/F concentration (55,200 pg/g, dry weight in a subsurface layer) was found in sediment collected at the mouth of the Saginaw River. Concentrations of PCDFs were greater than the concentrations of PCDDs in sediment from the Saginaw River. 2,3,7,8-TCDF, 1,2,3,7,8-PeCDF, and 2,3,4,7,8-PeCDF were the major PCDF congeners found in sediments from the Saginaw River. The elevated concentrations of PCDFs, and the predominance of the less highly chlorinated PCDF congeners, in sediments from the Saginaw River were similar to previously determined characteristics of the PCDF contamination of the Tittabawassee River, another tributary of the Saginaw River. These results suggest the existence of a major source of PCDFs within the watershed. A few localized areas of high PCDD/F and PCB concentrations, with unique congener compositions, in the Saginaw River indicated the presence of other minor sources, such as wastewater treatment plants. PCDFs were the major contributors to TEQs in sediment and soils from the Saginaw River and Saginaw Bay. Approximately 30% of the samples analyzed in this study had values exceeding the screening level of 50 pg TEQ/g, dry wt, suggested for soils by the Agency for Toxic Substances and Disease Registry. PMID- 17917762 TI - Historical distribution of PCDDs, PCDFs, and coplanar PCBs in sediment core of Ariake Bay, Japan. AB - Persistent organic pollutants, particularly polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), coplanar polychlorinated biphenyls (co-PCBs) and toxic equivalent quantity (pg TEQ g(-1)) were determined in sediment samples collected from Ariake Bay, Japan. The total concentration of PCDDs/DFs in surface sediment of the Yabe River (17,000 pg g(-1) dry wt) was approximately 2.5-fold higher than that of the Chikugo River (7,000 pg g(-1) dry wt) which has the highest flow amount of about 40% inputting to the northern Bay. The congener profile suggested that dioxin sources could be close to the study area. The PCDDs concentration (12,000 pg g(-1) dry wt) in the sediment layer, which was corresponding to the 1960s, was reflected on the pentachlorophenol (PCP) contamination occurring at that time in Ariake Bay. The reflecting of PCP in the sediment core was significantly considered by hierarchical cluster analysis. Based on isomer-specific analysis, the tetrachlorodibenzo-p-dioxin composition showed a gradual increase from 1978-1982 toward the surface, indicating that the pollution source at that period might be more influenced by chloronitrofen than PCP. This fact was supported, based on the ratio of Sigma PCDD to Sigma PCDF concentration. The PCDDs/DFs-derived TEQ contributed more than 90% of the SigmaTEQ (PCDDs/DFs and co-PCBs) in all the detected sediment layers. The contribution of PCBs to the total TEQ was low; however, an increasing historical trend of concentration was clearly observed. PMID- 17917763 TI - The influence of biomass on the toxicity of hydrophobic organic contaminants. AB - Hydrophobic organic contaminants (HOCs) enter the marine environment through several means, including industrial, urban, and agricultural runoff, and accumulate in sediments. Methods for measurement of sediment toxicity include porewater tests using sea urchin (Arbacia punctulata) fertilization and embryological development assessments. Previous studies investigating sediments from Boston Harbor determined that significant binding of contaminants to organic matter led to insufficient evidence of the bioavailability of HOCs in porewater toxicity tests. It was hypothesized that excessive biomass in testing systems prevents a critical body residue of HOCs from forming, thus curbing toxic effects. In this study, the effect of biomass on the toxicity of phenanthrene (a polycyclic aromatic hydrocarbon) and lindane (an organochlorine pesticide) were assessed individually and combined in a mixture. The fertilization toxicity test for phenanthrene and mixture solutions containing both compounds revealed less biomass in the test vial caused higher toxicity levels, the fact of which was enhanced with increased hydrophobicity. The 50% inhibition concentration (IC(50)) of phenanthrene to sea urchin fertilization success in test vials with 50 eggs/mL (lowest biomass concentration tested) was 3.72 micromol/L, but in vials with 100 to 400 eggs/mL, the IC(50) was >4.12 micromol/L. Toxicity of several concentrations of the phenanthrene and lindane mixture to sea urchin fertilization success and embryological development was significantly higher at lower biomasses (50 and 100 eggs or embryos/mL) than with biomasses > or =200 eggs or embryos/mL. The results suggest that when testing environmental samples that may contain HOCs, lowering the biomass can help better estimate sediment toxicity using porewater tests. PMID- 17917764 TI - Competition between native flow and graft flow after coronary artery bypass grafting. Impact on indications for coronary artery bypass grafting for localized stenosis with giant aneurysms due to Kawasaki disease. AB - We report the postoperative course of native and graft flow after coronary artery bypass grafting (CABG) in two patients with giant aneurysms and localized stenosis due to Kawasaki disease (KD). Although both patients had undergone CABG to the left anterior descending artery (LAD) with the left internal thoracic artery (ITA), at 5 and 10 years old, respectively, the ITA grafts were occluded 1 month postsurgery. However, when the two patients suffered complete occlusion of the native LAD more than 10 years after surgery, angiograms showed that the ITA grafts had reopened. We believe that this postoperative course reflects competition between the native artery flow and graft flow after CABG. CABG in patients with severely delayed coronary flows or recurrence of thrombus in giant aneurysms was ineffective in preventing myocardial infarction or damage. We conclude that CABG in giant aneurysm without significant localized stenosis should be avoided. PMID- 17917765 TI - Modeling the Fontan circulation: where we are and where we need to go. AB - The Fontan procedure and its subsequent modifications over the past 30 years can be described as a class of surgical procedures for patients born with complex congenital heart disease exhibiting a single-ventricle physiology. The long-term outcome for children currently undergoing a Fontan procedure remains worrisome because of multiple late morbidities observed. Despite significant modeling efforts spanning three decades, improvements to the Fontan procedure have occurred without comprehensive validation from these modeling studies. Careful examination shows that modeling studies to date offer only a "glimpse through a keyhole" into understanding and modeling a representative range of the variations in anatomy and physiology that exist in Fontan patients. Suggestions for future investigations are provided. PMID- 17917767 TI - QT dispersion and diastolic functions in differential diagnosis of primary mitral valve prolapse and rheumatic mitral valve prolapse. AB - There is no specific criteria established to guide physicians in the differential diagnosis of primary mitral valve prolapse (MVP) and rheumatic MVP. Previous studies suggested that history and pathology of mitral valve could be helpful in differential diagnosis of MVP. The aim of this study was to evaluate the value of QT interval, QT dispersion, and diastolic function in differential diagnosis of MVP. We examined electrocardiographies and echocardiographies of 24 primary MVP patients, 20 rheumatic MVP patients, and 21 healthy subjects. MVP was defined as superior displacement of the mitral leaflets more than 2 mm into the left atrium during systole. QT dispersion was significantly higher in primary MVP patients (71 +/- 13.5 ms, p < 0.01). Maximum QT dispersion value in rheumatic MVP patients was 55 ms. E and A velocity values which show ventricular early and late diastolic filling, were lower in primary MVP patients (p < 0.01). There was no difference in the heart rate corrected QT interval values between the primary MVP patients (397 +/- 28.1), rheumatic MVP patients (403 +/- 23.8) and healthy children (404 +/- 15.8; p > 0.05). Our findings may indicate that QT dispersion can be used as a parameter for differential diagnosis of primary MVP and rheumatic MVP. Further studies are needed to identify a cut-off point of QT dispersion. PMID- 17917766 TI - Methylenetetrahydrofolate reductase TT genotype as a predictor of cardiovascular risk in hypertensive adolescents. AB - Methylenetetrahydrofolate reductase (MTHFR) is associated with homocysteine level. In deficit of MTHFR, cardiovascular risk is increased with hyperhomocysteinemia and hypomethionemia. Mutation of the MTHFR gene is associated with the risk for premature cardiovascular diseases. However, the association between MTHFR mutation and cardiovascular risk is still controversial. The purposes of this study were to determine whether MTHFR genotype is associated with cardiovascular risks in hypertensive adolescents and to investigate the association between MTHFR genotype and carotid intima-media thickness (IMT). Forty-three hypertensive adolescents were included in this study. Serum lipid levels, insulin, vitamin B(12), folate, renin, aldosterone, angiotensin converting enzyme, and homocysteine levels were evaluated. The carotid IMT and diameter were estimated by ultrasound. Brachial-ankle pulse wave velocity was also measured. Polymerase chain reaction was conducted to amplify genomic DNA fragment containing C677T position of the MTHFR gene. The height, weight, body mass index, obesity index, arm circumference, fat mass, and fat distribution were significantly greater in patients with C677T mutation. The C677T mutation group showed significantly greater carotid IMT, higher homocysteine, and lower folic acid levels than the normal genotype group. Interpretation of MTHFR genotype might be useful in predicting the development of premature coronary artery disease in hypertensive adolescents. PMID- 17917768 TI - Magnetic resonance evaluation of criss-cross heart. AB - The purpose of this study was to report 18 cases of criss-cross heart and to evaluate the utility of magnetic resonance imaging (MRI) for the determination of atrioventricular segmental situs and atrioventricular alignment in patients with criss-cross heart. From August 1999 to March 2007, 18 consecutive patients with criss-cross heart were studied using a 1.5T MR scanner. Echocardiography and x ray angiocardiography were performed in all patients and surgery was performed in 12 patients. MR examination resulted in the same diagnosis with x-ray angiocardiography in 94.4% of patients (17 of 18). The visceroatrial situs was solitus in 17 patients with normal left side heart position in 14 and dextrocardia in 3. The visceroatrial situs was inversus in 1 patient with dextrocardia. The atrioventricular connections were concordant in 17 patients and discordant in 1. In all patients, the right ventricle was located superiorly and the left ventricle inferiorly. Criss-cross heart is a rare malformation caused by abnormal rotation of ventricles in the embryo. Diagnosis of criss-cross heart is difficult and MRI was very helpful. MRI allowed clear visualization of the horizontal ventricular septum, the crossing inflow streams, atrioventricular segmental situs, atrioventricular alignment, and other malformations. PMID- 17917769 TI - Coronary involvement in infants with Kawasaki disease treated with intravenous gamma-globulin. AB - AIM: To analyze the clinical spectrum and the incidence of coronary involvement in infants with typical Kawasaki's disease (KD). PATIENTS AND METHODS: A retrospective study was performed on children one year of age or younger diagnosed from February 1992 to January 2006 with typical KD. Children with incomplete forms of the disease were not included. RESULTS: Twenty-five infants were diagnosed with KD during the study period. The median age of the patients was 10 months (range, 4-12 months). All children but one received intravenous gammaglobulin (IVIG), 84% before the 10th day of disease. Seven patients (28%) required the administration of more than one dose of IVIG, because persistence of fever. Coronary artery disease (CAD) was recorded in 6 cases (24%), five of them being boys. All patients with CAD were treated with ASA plus IVIG and 84% of them received this therapy within the first 10 days of the KD onset. CONCLUSIONS: In spite of the exclusion of our study of incomplete presentations and of an early administration of IVIG in our patients, we have observed a high rate of infants who developed CAD, which is similar to the one reported in children who do not receive IVIG. PMID- 17917770 TI - Habitat acquisition strategies for grassland birds in an urbanizing landscape. AB - Habitat protection for grassland birds is an important component of open space land acquisition in suburban Chicago. We use optimization decision models to develop recommendations for land protection and analyze tradeoffs between alternative goals. One goal is to acquire (and restore if necessary) as much grassland habitat as possible for a given budget. Because a viable habitat for grassland birds consists of a relatively large core area with additional parcels of grassland habitat nearby, the second goal is to minimize total pairwise distance between newly protected parcels and large existing reserves. We also use the concept of an effective grassland habitat area, which considers influences that neighboring land covers have on grassland habitat suitability. We analyze how the parcels selected for protection change as total protected effective area is traded off against total distance. As area is weighted more heavily, the selected parcels are scattered and unconnected. As total distance is weighted more heavily, the selected parcels coalesce around core reserves but protect less area. The differences in selected parcels as we change the objective function weights are caused by the differences in price per unit of effective habitat area across parcels. Parcels located in close proximity to the existing cores have relatively high prices per hectare of effective grassland area as a consequence of high restoration costs and adverse influences from roads, urban areas and/or forestland. As a result, these parcels have lower priority for selection when the area objective is weighted more heavily for a given budget. PMID- 17917771 TI - Surgical site infection among women discharged with a drain in situ after breast cancer surgery. AB - BACKGROUND: There are scarce data on the factors associated with surgical site infection (SSI) among women who are discharged with a drain in situ after breast cancer surgery. The aim of this study was to determine the incidence and the factors associated with SSI in a center where women are routinely discharged with a drain in place. METHODS: A prospective cohort study included 354 women who underwent surgical treatment for breast cancer at a referral center in Rio de Janeiro, Brazil. RESULTS: SSI was diagnosed in 60 patients (17%) after a median follow-up of 17 days. Most infections were caused by Staphylococcus aureus. The probability of bacterial colonization of the drain was 33% on postoperative day (POD) 7 and rose to 80.8% up to the POD 14. In 83% of the cases of microbiologically documented infection, SSI was caused by the same bacterial species that had been previously isolated from the drainage fluid. In multivariate regression analyses, age 50 years (p < 0.001), skin flap necrosis (p < 0.001), and bacterial colonization of the drain (p = 0.03) were independently associated with a higher incidence of SSI. CONCLUSION: The incidence of SSI among women who were routinely discharged with a drain in place was high. Older age, skin flap necrosis, and bacterial colonization of the drain were independent predictors of SSI. Modifications of the surgical technique aimed at reducing the risk of wound necrosis and early removal of the drain may contribute to lowering the risk of SSI among these patients. PMID- 17917772 TI - Role of fibrin glue as a sealant to esophageal anastomosis in cases of congenital esophageal atresia with tracheoesophageal fistula. AB - OBJECTIVE: The aim of this study was to characterize a successful approach for the management of infants with long-gap esophageal atresia (EA) with tracheoesophageal fistula (TEF). The goal was to preserve the native esophagus and minimize the incidence of esophageal anastomotic leaks using fibrin glue as a sealant over the esophageal anastomosis. METHOD: A total of 52 patients were evaluated in this study. Only patients in whom, gap between the two ends of the esophagus was > or = 2 cm were selected during January 2005 to January 2007. Patients were divided in two groups on the basis of block randomization. Group A comprised the patients in whom fibrin sealant was used as reinforcement on a primary end-to-end esophageal anastomosis; in group B, fibrin glue was not used. The two groups were compared in terms of esophageal anastomotic leak (EL), postoperative esophageal stricture (ES), and mortality. The statistical analysis was done using Fisher's exact test and the chi-squared test. RESULT: The number of anastomotic leaks in group A (glue group) was about one-fifth that in group B (no glue group). The incidence of ES was almost twice as high in group B as in group A. The mortality rate was almost threefold higher in group B (no-glue group). The higher incidence of EL and ES in group B compared to group A was statistically significant. CONCLUSION: Thus, fibrin glue when used as an adjunct to esophageal anastomosis for primary repair of long-gap EA with TEF appears safe in the clinical setting and may lower the chances of esophageal leak and anastomosis-site strictures. Hence, it can diminish the mortality and morbidity of these patients. PMID- 17917773 TI - Sequential preoperative ipsilateral portal and arterial embolization in patients with liver tumors: is it really the best approach? PMID- 17917774 TI - Transfusion alert for patients with curable cancer. AB - BACKGROUND: Perioperative transfusion (POT) is a negative prognostic factor for outcome in cancer patients undergoing surgery, and splenectomy (SPL) has been suggested to help obviate this negative effect. However, the prognostic independence among related factors and clinical significance of the interaction remains elusive. PATIENTS AND METHODS: Between January 1, 1990 and January 31, 2005, a total of 200 patients with proximal gastric cancer classified as having stage IB to III disease after curative resection were enrolled at Kitasato University Hospital in a retrospective manner, according to a previous report by Weitz et al. FINDINGS: POT was reconfirmed to be a potent univariate prognostic predictor in this model (p < 0.0001). The tendency was further augmented in patients without SPL, suggesting that SPL can counteract the negative aspects of POT. A multivariate analysis including related factors revealed that POT was one of the most potent independent prognostic factors, as were stage and age. Stratification analysis following the JCGC staging system revealed that the outstanding prognostic difference according to POT was recognized in patients anticipated to be cured, such as those with stage IB (p = 0.012) or II (p = 0.003) but not stage III. SPL could dramatically improve the prognosis with POT in stage IB patients but modestly if at all in those with stage II or III. INTERPRETATION: Our findings support the need for a clinical alert against POT throughout the course of cancer treatment, especially for curable disease; thus, POT should be carefully considered from an oncologic point of view. Appropriate strategies to minimize POT should be urgently developed. PMID- 17917775 TI - Liver resection and transplantation in the management of iatrogenic biliary injury. AB - BACKGROUND: Biliary injury during cholecystectomy can be managed successfully by biliary reconstruction in the majority of patients; however, a proportion of patients may require hepatic resection or even liver transplantation. METHODS: Data on all patients referred with biliary injuries were recorded prospectively. The details of patients who required hepatic resection or transplantation were analyzed and compared to those patients managed with biliary reconstruction alone. RESULTS: From November 1984 until November 2003 there were 119 patients referred with Strasberg grade E injuries to the biliary tree, 14 of whom (9 women, 5 men) required hepatic resection or transplantation. The median age of these 14 patients was 48 (range: 30-81) years. Nine patients were considered for hepatic resection, and of these six underwent right hepatectomy, two had a left lateral sectionectomy, and one patient was deemed unfit for surgery and underwent metal stenting of the right hepatic duct. All patients are alive and remain well. Five patients developed hepatic failure and were considered for liver transplantation. Two patients who were unfit for transplantation died, and another died while on the waiting list for transplantation. The remaining two patients underwent liver transplantation, and one of them died from overwhelming sepsis. Concomitant vascular injury was demonstrated in 8 of the 14 patients (57%), and in 3 of the 4 (75%) patients that died. CONCLUSIONS: Hepatic atrophy or sepsis after biliary injury can be managed successfully with hepatic resection. Liver transplantation is required occasionally for patients with secondary biliary cirrhosis, but is rarely successful for early hepatic failure following iatrogenic biliary injury. PMID- 17917776 TI - On the 100th anniversary of sterile catgut kuhn: Franz Kuhn (1866-1929) and the epistemology of catgut sterilization. AB - The long road to effective catgut sterilization began with the work of Lord Joseph Lister (1867) and did not end until 40 years later. At the end of the nineteenth century dozens of different techniques were used to "sterilize" catgut, by immersing the cord in a cold chemical solution, by exposing it to steam, or by a combination of the two techniques, yet none of these approaches offered the ultimate solution. One of the many physicians working on the catgut problem at that time was the German surgeon Franz Kuhn (1866-1929), best known as a pioneer of intubation anesthesia. This review offers a brief biographical sketch of Kuhn's life and career on the occasion of the centenary of Sterile Catgut Kuhn. The goal of the present study is to describe several landmarks in the development of the catgut sterilization method. To explain this process, two approaches are taken: first, an analysis to see whether the character traits of the typical surgeon at that time provided the soil in which innovation could thrive, and second, an epistemological examination of the conceptual models for the attainment of knowledge current at that time. Perspectives for the future are explored in light of the "imperative of responsibility" of Hans Jonas. PMID- 17917778 TI - Respiratory muscle strength in the physically active elderly. AB - Advancing age is associated with a decline in the strength of the skeletal muscles, including those of respiration. Respiratory muscles can be strengthened with nonrespiratory activities. We therefore hypothesized that regular exercise in the elderly would attenuate this age-related decline in respiratory muscle strength. Twenty-four healthy subjects older than 65 years were recruited (11 males and 13 females). A comprehensive physical activity survey was administered, and subjects were categorized as active (n = 12) or inactive (n = 12). Each subject underwent testing of maximum inspiratory and expiratory pressures (PI(max) and PE(max)). Diaphragmatic thickness (tdi) was measured via two dimensional B-mode ultrasound. There were no significant differences between the active and inactive groups with respect to age (75 vs. 73 years) or body weight (69.1vs. 69.9 kg). There were more women (9) than men (3) in the inactive group. Diaphragm thickness was greater in the active group (0.31 +/- 0.06 cm vs. 0.25 +/ 0.04 cm; p = 0.011). PE(max) and PI(max) were also greater in the active group (130 +/- 44 cm H(2)O vs. 80 +/- 24 cm H(2)O; p = 0.002; and 99 +/- 32 cm H(2)O vs. 75 +/- 14 cm H(2)O; p = 0.03). There was a positive association between PI(max )and tdi (r = 0.43, p = 0.03). Regular exercise was positively associated with diaphragm muscle thickness in this cohort. As PE(max) was higher in the active group, we postulate that recruitment of the diaphragm and abdominal muscles during nonrespiratory activities may be the source of this training effect. PMID- 17917777 TI - Identification of differentially expressed genes from Rhodothermus sp. XMH10 in response to low temperature using random arbitrarily primed PCR. AB - Most research on the adaptation of thermophiles is focused on their adaptation to heat stress; only a few studies are focused on their cold adaptation. In this report, the thermophilic bacterium Rhodothermus sp. XMH10 was examined to gain a better understanding of gene expression in response to low temperature. Random arbitrarily primed polymerase chain reaction (RAP-PCR) was used to isolate and identify differentially expressed genes of bacteria grown at 45 degrees C (lowest) compared to those at 75 degrees C (optimal). Fifty-three differential cDNA fragments in total were isolated. Among them, 35 different cDNAs were analyzed by Northern blot, and 17 were confirmed to be differentially expressed at the transcriptional levels. These genes reflected a profile of differential expression and were involved in many physiological processes such as metabolism, cell membrane alterations, and regulatory adaptive response; most of them have never been previously reported. This study provides some new information on the adaptation of thermophilic bacteria to environmental temperature stress. PMID- 17917780 TI - Benefits of using the DCM4CHE DICOM archive. PMID- 17917779 TI - Real-time polymerase chain reaction assays for rapid detection and quantification of Noctiluca scintillans zoospore. AB - Application and availability of real-time polymerase chain reaction (PCR) assay to detect and quantify the Noctiluca scintillans zoospore were investigated seasonally. Specific primer set for N. scintillans 18S rDNA was designed and applied to real-time PCR assay using the serial dilutions of N. scintillans zoospores. The real-time PCR assays with Ns63F and Ns260R primers were applied to sea water samples collected weekly in Manazuru Port of Sagami Bay, Japan from April 2005 to June 2006. We developed effective DNA preparation steps for collecting the template DNA of N. scintillans zoospore: size fraction and filter concentration of the water samples, fixation with Lugol solution, cell lysis, and purification. This method is useful for the monitoring of the zoospores of N. scintillans, and can also be used for other small and physiologically fragile planktonic cell. Variation in the density of zoospore was successfully detected in the field samples. The peak density of N. scintillans zoospore was observed to occur just before or at the same time as the peak of the vegetative cells. Moreover, zoospores were detected in seawater even when the vegetative cells were not observed. The presence of zoospore was found all year round in the present study. In this regards, this information is essential for the study of the life cycle and seasonal variation of N. scintillans in the coastal waters. PMID- 17917781 TI - Recombinant factor VIIa: a useful tool for life-threatening colonic bleeding. Report of a case. PMID- 17917782 TI - L1 is associated with favorable outcome in neuroblastomas in contrast to adult tumors. AB - BACKGROUND: Neuroblastoma is the most common solid tumor in childhood with unconventional clinical behavior. L1, a neuronal cell adhesion molecule, is associated with poor survival in malignant adult tumors. The aim of the current study was to determine expression of L1 in pediatric neuroblastoma. METHODS: L1 expression was assessed on a tissue microarray with 66 surgically resected neuroblastoma samples by immunohistochemistry with a monoclonal antibody and peroxidase method. Additionally, mRNA expression was analyzed by reverse transcriptase-polymerase chain reaction with L1-specific primers. Data were correlated survival data by log rank test and Cox regression multivariate analysis. RESULTS: L1 was detected in 57 (86%) of 66 neuroblastomas, whereas 9 (14%) were L1 negative. Median survival of all children was 72 months. Analysis with Kaplan-Meier method revealed a surprising and contrary finding to adult tumor entities: an association of L1 positivity with better event-free and overall survival (P < .001 and P < .01 by log rank test). Multivariate Cox regression analysis showed an independent prognostic impact of L1 negativity for event-free and overall survival of the children (P < .05). CONCLUSIONS: In contrast to adult tumor entities, where L1 is associated with aggressive clinical behavior, our data show that L1 predicts good outcome in children with neuroblastoma. This novel finding suggests an inverse role of L1 in neuroblastoma. Future studies might focus on the molecular basis of the varying effect of L1 in different tumors. PMID- 17917783 TI - Cyanotic congenital heart disease (CCHD) with symptomatic erythrocytosis. AB - Secondary erythrocytosis of cyanotic congenital heart disease (CCHD) is pathologically different from primary erythrocytosis of polycythemia vera (PV). An association between elevated hematocrit and thrombosis has been established in PV patients, and treatment guidelines recommend maintaining hematocrit <45%. Although an association between elevated hematocrit and thrombosis has not been established in CCHD and secondary erythrocytosis, the current clinical practice is to phlebotomize these patients to hematocrit <65%. We report a 21-year-old woman with CCHD who presented with symptomatic erythrocytosis with numbness and tingling with hemoglobin 25.2 g/dl and hematocrit 75.8%. Her symptoms resolved with IV hydration. Other factors, including dehydration and iron deficiency, may precipitate hyperviscosity symptoms. The treatment is volume replacement and low dose iron therapy, not phlebotomy. Repeated phlebotomy causes iron deficiency with microcytic erythrocytes, which increases the whole blood viscosity and, therefore, can potentially accentuate rather than decrease the risk for a cerebrovascular accident. PMID- 17917785 TI - Stapled hemorrhoidopexy: a prospective study from pathology to clinical outcome. AB - Stapled hemorrhoidopexy is widely accepted to treat hemorrhoids, but serious complications have been reported. In this prospective audit, we correlated clinical outcome with pathological findings. From January 2003 to April 2007, 94 patients underwent hemorrhoidopexy. Macroscopic appearance of the specimen (shape, size, and depth) was recorded. Microscopically, the presence of columnar, transitional, and squamous epithelium, the involvement of circular/longitudinal smooth muscle, and features of mucosal prolapse were assessed. Clinical outcome was evaluated by a validated questionnaire. Postoperative pain, secretion, and bleeding durations were 12.7+/-10.6, 5.6+/-9.6, and 6.3+/-8.4 days. Patient's return to work averaged 16.7+/-10.7 days. Fissure, skin tags, and anal strictures were observed in 23.4%. Seven patients experienced pain for a significantly longer period of time. All specimens contained columnar mucosa, but 29.8% contained columnar and transitional epithelium and 12.8% contained columnar, anal transitional, and stratified squamous epithelium. Smooth muscle was observed in 62.7%. Pain was significantly increased if transitional epithelium was present in the specimen. No correlation or differences were observed if smooth muscle was present, although postoperative bleeding was more frequent. Hemorrhoidopexy is safe and effective. The specimen should always be sent for pathology examination. Only columnar epithelium should be present and, although the presence of smooth muscle does not influence the outcome in terms of functional results, its presence may play a role in postoperative bleeding. PMID- 17917784 TI - CT vs MRCP: optimal classification of IPMN type and extent. AB - INTRODUCTION: Intraductal papillary mucinous neoplasms (IPMNs) of the pancreas are being diagnosed with increased frequency. CT scanning commonly serves as the primary imaging modality before surgery. We hypothesized MRCP provides better characterization of IPMN type/extent, which more closely matches actual pathology. METHODS: Of 214 patients treated with IPMN (1991-2006), 30 had both preoperative CT and MRCP. Of these, 18 met imaging study criteria. Independent readers performed retrospective, blinded analyses using standardized criteria for IPMN type and extent. RESULTS: A ductal connection was detected on 73% of MRCP scans and only 18% of CT. IPMN type was classified differently in seven (39%); four (22%) of which were read on CT as having main duct involvement where this was not appreciated on MRCP or found on surgical pathology. MRCP showed multifocal disease in 13(72%) versus only 9(50%) on CT. A different disease distribution was seen in 9(50%). Finally, 101 branch lesions were identified on MRCP compared to 46 on CT. CONCLUSIONS: CT falls short of MRCP in detecting a ductal connection, estimating main duct involvement, and identification of small branch duct cysts. These factors influence diagnostic accuracy, cancer risk stratification and operative strategy. MRCP should be employed for optimal management of patients with IPMN. PMID- 17917786 TI - Efficacy of radical surgery in preventing early local recurrence and cavity related complications in hydatic liver disease. AB - BACKGROUND: Hydatic disease of the liver remains to be a complex worldwide problem especially in rural areas. Early local recurrence and cavity-related complications are still a matter of conflict in the management of hydatic liver disease. The aim of this study is to investigate efficacy of the type of surgical treatment in preventing early local recurrence and cavity-related complications of this disease. Here, we present the preliminary results of our study. METHODS: This study was performed prospectively including 32 patients who were operated for hydatic liver disease between January 2001 and January 2005. Patients were randomized into radical and conservative surgery groups. Recurrences at the primary surgical site in the first 2 years were considered as early local recurrence and biliary leakage, biliary fistula, cavity abscess, etc. were considered as cavity-related complications. RESULTS: Early local recurrences were observed only after conservative surgical procedures (p=0.045). Recurrent cysts were found to be due to satellite cysts or pericystic disease. Cavity-related complications were seen in six patients in the conservative surgery group (p=0.011). CONCLUSIONS: In suitable patients, radical surgical resection provides an effective surgical management option in preventing early local recurrence and cavity-related complications when compared to conservative surgical approaches. PMID- 17917787 TI - Calculation of pigment transition energies in the FMO protein: from simplicity to complexity and back. AB - The Fenna-Matthews-Olson (FMO) protein of green sulfur bacteria represents an important model protein for the study of elementary pigment-protein couplings. We have previously used a simple approach [Adolphs and Renger (2006) Biophys J 91:2778-2797] to study the shift in local transition energies (site energies) of the FMO protein of Prosthecochloris aestuarii by charged amino acid residues, assuming a standard protonation pattern of the titratable groups. Recently, we have found strong evidence that besides the charged amino acids also the neutral charge density of the protein is important, by applying a combined quantum chemical/electrostatic approach [Muh et al. (2007) Proc Natl Acad Sci USA, in press]. Here, we extract the essential parts from this sophisticated method to obtain a relatively simple method again. It is shown that the main contribution to the site energy shifts is due to charge density coupling (CDC) between the pigments and their pigment, protein and water surroundings and that polarization effects for qualitative considerations can be approximated by screening the Coulomb coupling by an effective dielectric constant. PMID- 17917788 TI - Osteoma of the calvaria in L-2-hydroxyglutaric aciduria. AB - L-2-Hydroxyglutaric aciduria (L-2-OHGA) is a rare autosomal recessive neurometabolic disease linked to chromosome 14q21.1 and is caused by mutations in the gene that most likely encodes L: -2-hydroxyglutarate dehydrogenase, which normally catalyses L: -2-hydroxyglutarate to alpha-ketoglutarate. It is characterized by progressive mental deterioration, pyramidal and cerebellar syndromes, macrocephaly and marked polycystic white-matter degeneration mainly involving frontal lobes. Brain tumours of variable nature have frequently been observed in L-2-OHGA. We report a patient affected by this disease who at the age of 20 years developed a bone tumour involving the right frontal region of the calvaria. He had first presented at the age of 10 years with psychomotor delay, clumsy gait and moderate mental impairment. Examination showed macrocephaly, cerebellar ataxia and quadripyramidal syndrome. Brain MRI showed low signal intensities on T1-weighted images and high signal intensities on T2-weighted images in cerebral subcortical white matter. Serum and urinary amino acid assay was normal. Urinary 2-hydroxyglutaric acid was 1418 mmol/mol creatinine (controls <25). Analysis of the L-2-hydroxyglutarate dehydrogenase gene revealed a homozygous mutation in exon 2 (A320G). At the age of 20 years, an osteoma of the right frontal bone was diagnosed. This finding reinforces the opinion concerning the association of L-2-OHGA and tumorigenesis and prompted us to verify the possible responsibility of some overproduced substances in this disease for the development of tumours and to look for any correlation between the type of mutation in the L-2-OHGA gene and the tumorigenic potential observed in some patients affected by this disease. PMID- 17917789 TI - Combined effects of the angiogenic genes polymorphisms on prostate cancer susceptibility and aggressiveness. AB - The single-gene approaches in association studies of polygenic diseases are likely to provide limited value in predicting risk. The combined analysis of genetic variants that interact in the same pathway may amplify the effects of individual polymorphisms and enhance the predictive power. To evaluate higher order gene-gene interaction, we have examined the contribution of four angiogenic gene polymorphisms (VEGF-1154G/A; VEGF-634G/C; MMP9-1562C/T and TSP1-8831A/G) in combination to the risk of prostate cancer. For the combined analysis of VEGF and MMP9 SNPs, we found a significant gene-dosage effect for increasing numbers of potential high-risk genotypes. Compared to referent group (low-risk genotypes), individuals with one (OR = 2.79, P = 0.1), two (OR = 4.57, P = 0.02) and three high-risk genotypes (OR = 7.11, P = 0.01) had increasingly elevated risks of prostate cancer. Similarly, gene-gene interaction of VEGF and TSP1 polymorphisms increased risk of prostate cancer in additive manner (OR = 6.00, P = 0.03), although the TSP1 polymorphism itself was not associated with the risk. In addition, we examined the synergistic effect of these polymorphisms in relation to prostate cancer prognosis according to histopathological grade and clinical stage at diagnosis. Cross-classified analysis revealed potential higher order gene-gene interactions between VEGF and TSP1 polymorphisms in increasing the risk of developing an aggressive phenotype disease. Patients carrying three high-risk genotypes showed a 20-fold increased risk of high-grade tumor (OR = 20.75, P = 0.002). These results suggest that the gene-gene interaction of angiogenic gene polymorphisms' increased risk of prostate cancer onset and aggressiveness. PMID- 17917790 TI - The special location of p-H3 and p-CENP-A on heterochromatin during mitosis in MCF-7. AB - CENP-A locates at nucleosome as histone H3-like proteins, and is phosphorylated during mitosis. We investigated the dynamic distribution of p-CENP-A to explore the details of its function. We found that p-CENP-A was phosphorylated at late prophase, and the signal of p-CENP-A arranged at equatorial plate along with nucleosomes at metaphase, but moved to midbody at later phase of mitosis. The phosphorylation modification of CENP-A shares some characters of H3, but has different temporal patterns during mitosis. Our results suggested that the CENP-A might have similar functions as H3, but with different patterns for their different binging materials. PMID- 17917791 TI - Chronical illness and maternity: life conditions, quality of life and coping in women with multiple sclerosis. AB - AIM: Research on life circumstances, quality of life (QOL) and coping behavior in mothers with multiple sclerosis (MS). METHOD: Anonymous standardised questionnaire sent to 7,050 members of a section of the German MS Association (response rate 44.8%). Comparison of 482 mothers with MS (children aged < 18 years) with 607 childless women with MS. RESULTS: No statistically significant differences concerning age, MS course, complaints or number of exacerbations. Mothers with MS more frequently had a relationship, a higher monthly net income, were less employed, EDSS-score was lower and disease duration shorter. In QOL mothers with MS showed better social aspects even after multivariate adjustment for sociodemographic and disease-related variables. Influencing parameters on the social area of QOL were employment status, age, monthly household net income and disability. In their coping behavior mothers tended more to "religiosity/search for sense in life". CONCLUSION: There were several differences in sociodemographic data, QOL and coping behavior factors between mothers and childless women with MS. However, if motherhood itself has an influence on QOL and coping can not be derived from our data but there are some hints that motherhood seems to be no potential problem for living with the disease. Further research upon this topic is needed. PMID- 17917792 TI - Impact of smoking on asthma symptoms, healthcare resource use, and quality of life outcomes in adults with persistent asthma. AB - BACKGROUND: Smoking habits of asthmatics are similar to those of the general population. However, little attention has been paid to the associations between smoking and asthma-related outcomes. OBJECTIVE: To evaluate relationships between cigarette smoking, asthma symptoms, and asthma-related resource utilization in subjects with persistent asthma. METHOD: A stratified, random sample of adults from France, Germany, and the UK with persistent asthma were surveyed in 2001 through 2004. Statistical analyses compared asthma symptoms and healthcare resource utilization for cigarette smokers compared with those for non-smokers. RESULTS: Analyses included 1109 subjects with persistent asthma symptoms in 2001 2003 and 852 subjects with persistent asthma symptoms in 2004. Using multivariate analysis of data from eligible subjects in 2004 that did not have concomitant chronic obstructive pulmonary disease and adjusting for subjects' baseline and demographic characteristics, cigarette smokers were more likely to experience nighttime symptoms (OR 1.46, 95% CI 1.07, 1.97 P = 0.015) and were more likely to use healthcare resources than were non-smokers (P < or = 0.004). Findings were similar in a secondary analysis of subjects < or =55 years of age. CONCLUSIONS: Cigarette smoking appears to be associated with more asthma symptoms and more ED visits and hospitalizations in adults with persistent asthma. PMID- 17917793 TI - Response shift due to diagnosis and primary treatment of localized prostate cancer: a then-test and a vignette study. AB - AIM: Whether a prostate cancer diagnosis induces response shift has not been established so far. Therefore, we assessed response shift in men who were diagnosed with localized prostate cancer. PATIENTS AND METHODS: Out of 3,892 men who completed a questionnaire before screening, 82 were subsequently diagnosed with prostate cancer. Response shift was assessed in 52 (response 63%) by the then-test (EuroQol self-rating of health, Short-Form 36 mental health and vitality) and a novel method: rating of vignettes relating to side effects of prostate cancer treatment (urinary, bowel and erectile dysfunction). Three then tests were conducted: two referencing pre-diagnosis (measured pre- and post treatment), and one referencing pre-treatment (measured post-treatment). RESULTS: Then-test scores of pre-diagnosis health were significantly higher than original scores, indicating a more positive judgement in retrospect. Then-test scores of pre-treatment health were lower than original scores. Especially the vignette on erectile dysfunction was rated less bad after diagnosis versus before (P < 0.001, moderate effect size). CONCLUSIONS: We found evidence for response shift in men who were diagnosed with prostate cancer. Men evaluated urinary, bowel, and erectile dysfunction as less bad after they had become patients who can expect to experience these side effects. The rating of vignettes is a promising additional technique to assess response shift. PMID- 17917794 TI - Syntheses, characterization and fluorescent properties of six novel lanthanide complexes with N,N-diphenyl-2-(quinolin-8-yloxy)acetamide. AB - Six novel complexes of lanthanide nitrates (Ln=La, Sm, Eu, Gd, Tb, Dy) with a amide type ligand, N-methyl-N-phenyl-2-(quinolin-8-yloxy)acetamide (L) have been prepared and characterized by elemental analysis, conductivity measurements, IR and (1)H NMR spectra. The fluorescence properties of the complexes and the triplet state energy of the ligand were studied in detail. The result indicates that, the triplet state energy level of the ligand matches better to the resonance level of Eu(III) than Tb(III). In addition, the fluorescence intensities of the Eu(III) complex in different solutions(tetrahydrofuran, acetone and acetonitrile) are stronger than that in solid state. This is probably due to the solvate effect and the stoichiometry change of ligand with Eu(III) ion in solutions. PMID- 17917796 TI - Women with BRCA1 or BRCA2 mutations renegotiating a post-prophylactic mastectomy identity: self-image and self-disclosure. AB - The Facing Our Risk of Cancer Empowered (FORCE) website is devoted to women at risk for hereditary breast and ovarian cancers. One of the most frequently discussed topics on the archived messaged board has been prophylactic mastectomy (PM) for women with a BRCA1/2 mutation. We reviewed the messages, over a 4 year period, of 21 high risk women and their "conversational" partners who originally posted on a thread about genetic testing, genetic counseling and family history. We used a qualitative research inductive process involving close reading, coding and identification of recurrent patterns, relationships and processes in the data. The women sought emotional support, specific experiential knowledge and information from each other. They frequently found revealing their post PM status problematic because of possible negative reactions and adopted self-protective strategies of evasion and concealment outside of their web-based community. The FORCE message board was considered to be a safe place in which the women could be truthful about their choices and feelings. Results are discussed in terms of Goffman's concepts "stigma" and "disclosure" and Charmaz's concepts "interruptions," "intrusions" and a "dreaded future." PMID- 17917797 TI - Bafilomycin A1 is a potassium ionophore that impairs mitochondrial functions. AB - Novel activities of bafilomycin A1, a macrolide antibiotic known as an inhibitor of V-ATPases, were discovered. Bafilomycin A1 induced uptake of potassium ions by energized mitochondria and caused mitochondrial swelling, loss of membrane potential, uncoupling of oxidative phosphorylation, inhibition of the maximal respiration rates, and induced pyridine nucleotide oxidation. The mitochondrial effects provoked by nanomolar concentrations of bafilomycin A1 were connected to its activity as a potent, K(+)-specific ionophore. The K(+) ionophoric activity of bafilomycin A1 was observed also in black lipid membranes, indicating that it was an inherent property of the bafilomycin A1 molecule. It was found that bafilomycin A1 is a K(+) carrier but not a channel former. Bafilomycin A1 is the first and currently unique macrolide antibiotic with K(+) ionophoric properties. The novel properties of bafilomycin A1 may explain some of the biological effects of this plecomacrolide antibiotic, independent of V-ATPase inhibition. PMID- 17917799 TI - Effects of glycerol on the thermal dependence of the stability of human erythrocytes. AB - Incubation of human blood in saline solution of 0-36% (v/v) ethanol for 30 min produces lysis or stabilization of erythrocytes depending on the ethanol concentration. Under less elevated concentrations of ethanol, erythrocytes are present in expanded shapes (R state) that present lower stability and suffer lysis with increase in the ethanol concentration. Under more elevated concentrations of ethanol, erythrocytes are present in contracted shapes (T state) that have higher stability and suffer lysis at even more elevated ethanol concentrations. This work evaluated the effects of glycerol (0 to 2.0 M) and temperature (7 to 47 degrees C) on the stability of the R erythrocytes, characterized by the ethanol concentration at the mid-transition point (D (50R )) of the hemolysis curve (D (50R )). D (50R ) declined sigmoidally with increase in the glycerol concentration or temperature, due to transition of the R to the T state erythrocytes. In 1.5 M glycerol, the erythrocytes stability decreased below 32 but increased above 37 degrees C. The combination of temperature, glycerol and ethanol actions generates a critical value of osmotic pressure below which the R state predominates and above which the T state predominates. At 7 degrees C 1.5 M glycerol decreased the erythrocytes stability against ethanol but increased the erythrocytes stability against hypotonic shock. Those conditions favor the R state, which has a lower stability against ethanol; however, in the absence of ethanol, glycerol determines less water entrance in the erythrocytes, making more difficult its lysis by hypotonicity. PMID- 17917798 TI - Ischemic preconditioning enhances fatty acid-dependent mitochondrial uncoupling. AB - This study tests the hypothesis that ischemic preconditioning (IP) changes fatty acid (FA)-dependent uncoupling between mitochondrial respiration and oxidative phosphorylation. We found that IP does not alter mitochondrial membrane integrity or FA levels, but enhances membrane potential decreases when FA are present, in an ATP-sensitive manner. FA hydroperoxides had equal effects in control and preconditioned mitochondria, and GTP did not abrogate the IP effect, suggesting uncoupling proteins were not involved. Conversely, thiol reductants and atractyloside, which inhibits the adenine nucleotide translocator, eliminated the differences in responses to FA. Together, our results suggest that IP leads to thiol oxidation and activation of the adenine nucleotide translocator, resulting in enhanced FA transport and mild mitochondrial uncoupling. PMID- 17917800 TI - Behind the mask of method: political orientation and constitutional interpretive preferences. AB - Debate about how to best interpret the Constitution often revolves around interpretive methodologies (e.g., originalism or expansive interpretation). This article examines whether individuals' political orientation influences the methodologies they prefer to use to interpret the Constitution. We study this proposed relationship using a survey of federal law clerks and an experimental study with college students. The survey results indicate that, compared to conservatives, liberal clerks prefer the current meaning or the most plausible appealing meaning of the constitutional text, while conservatives prefer the original meaning of the text. Liberal clerks also prefer to interpret the Constitution much more expansively. The second study manipulates the policy implications of expansive interpretation and finds this manipulation differentially affects liberals' and conservatives' expansiveness preferences. PMID- 17917801 TI - Perceptions of prenatal testing for birth defects among rural Latinas. AB - Objectives To examine rural Latinas' understandings of prenatal testing, birth defects, and risk in the context of their expanded AFP (XAFP) screening decisions. Design We conducted a qualitative study using data from in-depth interviews with 33 Latina women receiving prenatal care at three clinic sites in rural areas of California. We analyzed qualitative data by identifying themes that emerged during iterative transcript readings. Quantitative data was used to generate descriptive summary statistics. Results The majority of the participants had not completed high school and had low levels of acculturation. Women in our study tended to view XAFP screening as a routine component of prenatal care that was important for the "health of the baby." Reasons for accepting the XAFP test included reassurance, emotional preparation, and desire for information. Misconceptions included a belief that a normal screening result provides a guarantee of the fetus's health. Generally, participants indicated that, regardless of the screening results, they would not undergo amniocentesis because of the potential miscarriage risk nor would they terminate a pregnancy if their fetus was found to have a chromosome problem. Numerous specific beliefs that differ substantially from medical models regarding birth defects were articulated. Conclusions XAFP screening decisions among rural Latinas are sometimes based on misconceptions of the meaning of test results and the risks associated with undergoing or foregoing testing, and therefore are frequently incongruent with personal testing philosophies and values. Educational efforts and counseling should include clear communication regarding the goals of prenatal screening programs to help these women make informed testing decisions that are reflective of their values and preferences. PMID- 17917803 TI - The early development of joint attention in infants with autistic disorder using home video observations and parental interview. AB - The aim in the current study was to investigate the early development of joint attention, eye contact and affect during the first 2 years of life, by using retrospective parental interviews and analyses of home videos of infants who were later diagnosed with Autistic Disorder (AD). The 36 children with AD and the 27 matched control children were all aged between 3 and 5 years at recruitment. Reported anomalies in gaze and affect emerged in the children with AD as early as the first 6 months of life, generally becoming more severe just prior to the second birthday. Video data confirmed these anomalies from as early as the first year. Joint attention impairments were found throughout the second year of life. The results suggest that early dyadic behaviours-eye contact and affect-may play a role in the joint attention impairment in AD. PMID- 17917802 TI - Distributed representation of perceptual categories in the auditory cortex. AB - Categorical perception is a process by which a continuous stimulus space is partitioned to represent discrete sensory events. Early experience has been shown to shape categorical perception and enlarge cortical representations of experienced stimuli in the sensory cortex. The present study examines the hypothesis that enlargement in cortical stimulus representations is a mechanism of categorical perception. Perceptual discrimination and identification behaviors were analyzed in model auditory cortices that incorporated sound exposure-induced plasticity effects. The model auditory cortex with over-representations of specific stimuli exhibited categorical perception behaviors for those specific stimuli. These results indicate that enlarged stimulus representations in the sensory cortex may be a mechanism for categorical perceptual learning. PMID- 17917805 TI - Did Hans Asperger (1906-1980) have Asperger syndrome? PMID- 17917806 TI - School climate and implementation of a preventive intervention. AB - Although there has been wide dissemination of research-based psychosocial prevention programs, a similarly strong research base to guide program implementation has been lacking. Program implementation has been particularly difficult for schools, due partly to insufficient understanding of how school ecologies interact with these programs. This study examined the effects of multiple dimensions of school climate on level and rate of change in implementation of a violence prevention intervention across three school years. Using multi-level modeling, the study found that teacher-reported support between staff and among teachers and students predicted higher average levels of implementation. Teacher-reported administrative leadership predicted greater growth in implementation across 3 years. Findings offer implications for an ecological model of program implementation that considers school-level contextual effects on adoption and sustainability of new programs in schools. PMID- 17917804 TI - Diagnostic validity of sensory over-responsivity: a review of the literature and case reports. AB - Atypical responses to sensory stimulation are frequently reported to co-occur with diagnoses such as autism, ADHD, and Fragile-X syndrome. It has also been suggested that children and adults may present with atypical sensory responses while failing to meet the criteria for other medical or psychological diagnoses. This may be particularly true for individuals with over-responsivity to sensation. This article reviews the literature related to sensory over responsivity and presents three pediatric cases that present a profile of having sensory over-responsivity without a co-occurring diagnosis. Findings from these cases provide very preliminary evidence to support the suggestion that sensory over-responsivity can occur as a sole diagnosis. Within this small group, tactile over-responsivity was the most common and pervasive form of this condition. PMID- 17917807 TI - Benign breast disease heterogeneity: association with histopathology, age, and ethnicity. AB - BACKGROUND: Benign breast biopsies with concurrent multiple benign lesions with different histopathologic diagnoses were termed heterogeneous benign breast disease (HBBD). Multiplicity of benign breast disease (BBD) lesions in a biopsy is a risk factor for progression to breast cancer (BC). Elucidation of the biological characteristics and clinical implications of HBBD may also be relevant to the refinement of risks for BC in women with a BBD diagnosis. DESIGN: In this study, we investigated the association of HBBD with histopathology, age, and ethnicity. A cohort of 4,341 women, 1,208 African Americans and 3,133 Caucasians, diagnosed with BBD, was identified after examination of an excisional breast biopsy. BBD biopsies were categorized as nonproliferative (NP, low risk or risk 1 lesions), proliferative without atypia (P, intermediate risk or risk 2 lesions), and proliferative with atypia (AH, high risk or risk 3 lesions). A BBD biopsy with only a single BBD lesion was termed simple BBD (SBBD). BBD biopsies with multiple lesions were further classified as single level HBBD (SL-HBBD) if the concurrent lesions were within the same risk level, or as multiple level HBBD (ML HBBD) if lesions fell into more than one risk group. RESULTS: In this cohort, 69% of women with a BBD diagnosis fit the HBBD criteria. Among women with HBBD, ML HBBD was almost three times more prevalent than SL-HBBD and was significantly more likely to be composed of risk 2 and risk 3 lesions. The likelihood of HBBD was 57% higher in Caucasian American women than in African American women with BBD (OR 1.57; 95% CI: 1.37, 1.81). The average lesion number in HBBD was directly proportional to increasing lesion risk (P < 0.001). Compared to women with risk 1 lesions, the likelihood of HBBD was 5.59 (95% CI: 4.85-6.44) and 17.0 (95% CI: 10.2-28.5) times higher when risk 2 and risk 3 lesions, respectively, were present. Women in the age range of 46-55 years and >55 years had a 3.12 (95% CI: 2.59, 3.75) and a 2.28 (95% CI: 1.94, 2.68) fold higher likelihood of HBBD compared to those < or =45 years. Significant interaction was found between concurrent lesion levels and age (P < 0.01). The likelihood of HBBD was considerably higher across all age groups for risk 3 lesions. Compared to the reference (risk 1, age < or =45), the likelihood of HBBD for risk 2 lesions was 4.4 times greater (95% CI: 3.70, 5.33) in women < or =45 YEARS, BUT THAT LIKELIHOOD INCREASED TO 17.6 (95% CI: 12.8, 24.2) AND 13.4 (95% CI: 10.1, 17.9) TIMES IN WOMEN OF 46-55 AND >55 YEARS, RESPECTIVELY. CONCLUSION: HBBD is more prevalent in Caucasian American women than in African American women. Women with higher risk BBD lesions are more likely to have HBBD. Lesion number and higher risk BBD lesions are significantly correlated with ML-HBBD. Additionally, the associations of HBBD and lesion risk level are modified by age. PMID- 17917809 TI - Introduction: exercise in patients with chronic heart failure. PMID- 17917808 TI - Vitamin supplement use and risk for breast cancer: the Shanghai Breast Cancer Study. AB - OBJECTIVE: The influence of vitamin supplements on breast cancer risk is unclear and the interactive effects of dietary and supplemental sources are unknown. This study investigated (1) the association between self-reported vitamin supplement use (multivitamin, A, B, C, and E) and breast cancer and (2) the combined effect of vitamin supplements in relation to dietary vitamin intakes on breast cancer risk. METHODS: The Shanghai Breast Cancer Study was a population-based case control study conducted in Shanghai in 1996-1998 (Phase I) and 2002-2004 (Phase II). Participants were aged 25-64 (Phase I) and 20-70 years (Phase II). The analyses included 3,454 incident breast cancer cases and 3,474 controls. Unconditional logistic regression models were used to determine adjusted odds ratios (ORs) for breast cancer risk associated with vitamin supplement use. RESULTS: Overall, breast cancer risk was not related to any vitamin supplement intake. However, a 20% reduction in breast cancer risk was observed with vitamin E supplement use among women with low-dietary vitamin E intake (OR = 0.8; 95% confidence interval (CI), 0.6-1.0). A non-significant 20% risk reduction was observed among vitamin B supplement users with low B dietary intake (OR = 0.8; 95% CI, 0.6-1.1). Frequent use of a vitamin B supplement was adversely associated with breast cancer risk among those with high dietary vitamin B intake (OR = 1.4; 95% CI: 0.9-2.1; P for interaction = 0.07). CONCLUSIONS: This study suggests that vitamins E and B supplements may confer protection against breast cancer among women who have low dietary intake of those vitamins. PMID- 17917810 TI - Biochemical characteristics of the herb mixture Prolipid as a plant food supplement and medicinal remedy. AB - Prolipid a known mixture of herbs is used as a plasma lipid lowering medicine. No side effects were registered. However, the bioactive substances of Prolipid were not investigated. Therefore in this investigation Prolipids bioactive compounds and antioxidant activity were studied. The contents of polyphenols and flavonoids were 19.87 +/- 2.09 and 3.09 +/- 0.31 mg gallic acid equivalent GAE/g DW and 2.09 +/- 0.24 and 0.57 +/- 0.05 mg catechin equivalent CE/g DW in water and methanol fractions, respectively. Anthocyanins (0.02 +/- 0.001 mg/g DW) and flavanols (7.58 +/- 0.81 microg CE/g DW) were found only in water fraction. The antioxidant activity of Prolipid, as determined by four different antioxidant assays [ferric reducing/antioxidant power (FRAP); cupric reducing antioxidant capacity (CUPRAC); trolox equivalent antioxidant capacity (TEAC); 1, 1-diphenyl-2-picrylhydrazyl radical (DPPH)], was higher in water than in methanol fraction. The correlation coefficients between polyphenols, flavonoids and antioxidant activities of Prolipid water extracts with TEAC were 0.97 and 0.90, respectively. It can be concluded that the content of polyphenol compounds in Prolipid is very high and they are the main contributors to Prolipids overall antioxidant activity. Prolipid is widely used in human treatment without known side effects on patients and is comparable to other medicinal plants, and as a strong antioxidant mixture could be used as a supplement to known atherosclerosis preventing diets. PMID- 17917811 TI - Five years follow-up of invasive prolactinomas with special reference to the control of cavernous sinus invasion. AB - BACKGROUND: Few data are presently available on the effective control of cavernous sinus (CS) invasion of invasive prolactinomas. The aim of this retrospective study, through a mean period of 5 years follow up, is to observe the tumor shrinkage of CS invasive prolactinomas, as well as PRL normalization with bromocriptine therapy. METHODS: 68 patients met the criteria of invasive prolactinomas (Grade III or IV in the classification scheme of Knosp and colleagues; serum PRL level greater than 200 ng/ml). 33 patients underwent bromocriptine therapy as the initial treatment, and 14 of these 33 had combined treatment with microsurgery and/or radiotherapy. The other 35 patients received microsurgery as the primary treatment, after which two patients had normal PRL without taking bromocriptine and other 33 patients received bromocriptine treatment after microsurgery. RESULTS: Tumor volume on magnetic resonance images had completely disappeared in 50 patients (74%), while all the other 18 patients had residual tumor in the parasellar areas, invading the CS, and 14 patients had a secondary empty sella due to tumor shrinkage. Of those 14 patients, seven still had elevated PRL levels; five had optic chiasmal herniation by different degrees (P < 0.05). There were 49 patients with normal PRL levels (72%); five patients with PRL levels more than 200 ng/ml. After the treatment, 14 patients with tumor volume disappearance on MR images and PRL normalization therefore withdrew from bromocriptine therapy. During a subsequent one-and-a-half-year follow-up, tumor recurrence and PRL increase were not found in those 14 patients. Twenty-seven patients maintained normal PRL levels with low-dose bromocriptine, of which 20 patients had their tumor disappear while seven patients had CS residual tumor. CONCLUSIONS: About three-fourths of prolactinomas with CS invasion can be effectively controlled not only with regard to tumor volume disappearance but also in serum PRL normalization. Residual tumor in the CS areas with PRL normalization and no pressure symptoms can be treated with low-dose of bromocriptine so as to achieve long-term tumor volume control and endocrine control. Great attention should be paid to CS residual tumors accompanying the empty sella, especially in cases with optic chiasmal herniation. PMID- 17917813 TI - Serum markers of cardiovascular risk in patients with acromegaly before and after six months of treatment with octreotide LAR. AB - Acromegaly is associated with increased morbidity and mortality from cardiovascular disease. Inflammatory markers, such as C-reactive protein and leucocyte count, haemostatic markers, such as fibrinogen and factor VIII and cardiac hypertrophy marker, B-type natriuretic peptide, have emerged as important cardiovascular risk markers in the general population. The objective of this study was to assess the serum levels of conventional, inflammatory, haemostatic markers and NT-pro BNP in mostly non-diabetic normotensive patients with acromegaly, as well as the effect of 6 months of octreotide LAR therapy on these markers. We studied 12 patients with active acromegaly, 12 patients in whom remission of acromegaly had been achieved by surgery and 12 healthy control subjects matched for age, gender and body mass index. At baseline fasting blood was obtained for measurements of GH, IGF-1, glucose, insulin, lipids, lipoprotein (a), C-reactive protein, leucocyte count, fibrinogen, factor VIII and NT-pro BNP. After baseline evaluation, patients with active acromegaly were treated with octreotide LAR for 24 weeks. At 24 weeks, measurements were repeated as on baseline. Insulin resistance index and fibrinogen levels were higher in patients with active acromegaly than patients and subjects in control groups. CRP, leucocyte count, factor VIII and NT-pro BNP were similar in the three groups. Octreotide LAR reduced GH, IGF-1 and insulin resistance index but did not alter levels of CRP and NT-pro BNP. PMID- 17917814 TI - Prevalence of indoor allergen exposures among New Orleans children with asthma. AB - Studies of inner-city asthmatic children have shown significant regional variation in dust allergen exposures. The home environment of asthmatic children in the Gulf South region of the USA has not been characterized. This study describes indoor dust allergen levels in the homes of 86 asthmatic children in New Orleans and explores regional variability in dust allergen exposure. Data were used from baseline home visits of children in the New Orleans Healthy Homes Initiative. Interview, visual observation, and environmental dust sampling data of 86 children between 4 and 17 years of age were analyzed. Seventy-seven percent of households had moderate (>2.0-9.9 microg/g) or high (> or =10.0 microg/g) levels of either Der p 1 or Der f 1 dust mite allergen and 56.6% had moderate (>2.0-8.0 U/g) or high (>8.0 U/g) levels of cockroach allergen (Bla g 1). The prevalence of high (>10 microg/g) levels of dog (Can f 1) allergen was 26.5%, and few households (6.0%) had high cat allergen (Fel d 1) levels (>8.0 microg/g). Households with average humidity levels >50% were three times more likely to have elevated dust mite levels (odds ratio = 3.2; 95% confidence interval = 1.1, 9.3; p = 0.03). Home ownership and education level were inversely associated with cockroach and dust mite allergen levels, respectively. Our findings reinforce the evidence of regional variability in dust allergen exposure levels. Asthmatic children living in the Gulf South are exposed to multiple indoor allergen exposures and live in a highly allergenic environment. PMID- 17917815 TI - Introducing HPV screening: challenges and fallacies. PMID- 17917812 TI - Collision lesions of the sella: co-existence of craniopharyngioma with gonadotroph adenoma and of Rathke's cleft cyst with corticotroph adenoma. AB - Collision lesions of the sellar region are relatively uncommon. Most contributions include a pituitary adenoma or a cyst/cystic tumor, particularly a Rathke cleft cyst. The association of craniopharyngioma with an adenoma is particularly rare. Among reported cases, some have included secondary prolactin cell hyperplasia due to pituitary stalk section effect. Herein, we report two collision lesions, including a gonadotroph adenoma with adamantinomatous craniopharyngioma and a corticotroph adenoma with Rathke's cleft cyst. Clinicopathologic correlation and a review of the literature are undertaken. PMID- 17917816 TI - Cardiovascular diseases attributable to hysterectomy: a population-based study. AB - BACKGROUND: Previous studies have confirmed an association between hysterectomy and an increased risk of cardiovascular diseases (CVDs), although some controversy remains. The aim of this study was to ascertain the attributable fraction (AF) of CVD due to hysterectomy. STUDY DESIGN: A cross-sectional study based on a random sample of 2,514 Finnish women aged 30-99 was carried out. Associations between hysterectomy and CVD were analysed using multivariate analyses. By considering hysterectomy as an indicator for risk of CVD rather than the cause of CVD, based on our results the AF and population attributable fraction (PAF) were calculated for conditions associated with hysterectomy. RESULTS: Some 33% of women aged 50 or older had undergone hysterectomy. In the univariate analyses, hysterectomy was significantly associated with hypertension, medication for hypertension, angina pectoris, stroke, age, education, oral contraceptive use, hormone replacement therapy (HRT), higher body mass index (BMI), fasting blood glucose and cholesterol. The fully-adjusted ORs for association between CVD and hysterectomy were dramatically lower than the crude ORs, and remained significant only for medication for hypertension. The proportion of hypertension among hysterectomised women that would be prevented if none had been hysterectomised (AF) was 19.6%, while the preventable proportion of hypertension among the female population that would be prevented if none had been hysterectomised (PAF) was 3.4%. For current use of medication the AF was 51.9% and the PAF was 13.5%. The AF of hysterectomy for stroke was 43.8% and the PAF was 10.2%. CONCLUSION: After adjusting for cardiovascular risk factors, we found no association between hysterectomy and ischemic heart diseases. The crude and age-adjusted associations found for some CVDs are most probably due to the more adverse initial risk profile of women who had undergone hysterectomy. The presented AF and PAF are related to indications of hysterectomy (such as uterine fibroids) rather than to the operation itself. There is a need to evaluate the cardiovascular risk factors of women with indications for hysterectomy. PMID- 17917817 TI - Utero-vaginal anastomosis in the treatment of cervical atresia. AB - Cervical atresia is an extremely rare malformation. Its incidence is not well known and management is controversial. The surgical technique and results of utero-vaginal anastomosis in 3 women affected by cervical atresia are described. PMID- 17917818 TI - Effects of estrogen deficiency on tooth movement after force application: an experimental study in ovariectomized rats. AB - OBJECTIVE: The aim of this study was to investigate the effects of estrogen deficiency on tooth movement in ovariectomized rats. MATERIAL AND METHODS: Forty two adult female Sprague-Dawley rats were assigned at random to one of the following groups: test group (n=20), ovariectomized rats (or estrogen-deficient rats); control group (n=22), non-ovariectomized rats. Two months after ovariectomy, expansion springs exerting 10 g of force were inserted between the upper central incisors in both groups. The amount of movement was measured daily until tooth movement began and then at intervals of 3 days. The rats were sacrificed 18 days after applying the expansion spring and histomorphometric analysis was performed along the left upper central incisor root towards the apex of the alveolar bone. RESULTS: The amount and speed of movement was observed to be greater in ovariectomized rats. On histomorphometric analysis, osteoblast and osteocyte counts on the pressure side were higher in the non-ovariectomized group than in the ovariectomized group (p<0.001). In contrast, the osteoclast count was significantly higher (p<0.01) in the ovariectomized group than in the non ovariectomized group. The osteoblast and osteocyte counts were significantly higher (p<0.001) on tension side in the non-ovariectomized group than in the ovariectomized group. CONCLUSION: Estrogen deficiency increased orthodontic tooth movements but counts of osteoblasts, which are responsible for new bone formation, were lower in regions of tension and of pressure. PMID- 17917819 TI - Improved physical fitness of cancer survivors: a randomised controlled trial comparing physical training with physical and cognitive-behavioural training. AB - We compared the effect of a group-based 12-week supervised exercise programme, i.e. aerobic and resistance exercise, and group sports, with that of the same programme combined with cognitive-behavioural training on physical fitness and activity of cancer survivors. One hundred and forty seven cancer survivors (all cancer types, medical treatment >or=3 months ago)were randomly assigned to physical training (PT, n=71) or PT plus cognitive-behavioural training (PT+CBT, n=76). Maximal aerobic capacity, muscle strength and physical activity were assessed at baseline and post-intervention. Analyses using multilevel linear mixed-effects models showed that cancer survivors' physical fitness increased significantly in PT and PT+CBT from baseline to post-intervention. Changes did not differ between PT and PT+CBT. Physical fitness of cancer survivors was improved following an intensive physical training programme. Adding a structured cognitive-behavioural intervention did not enhance the effect. PMID- 17917820 TI - Pancreatic ascites due to rupture of a mucinous cystic neoplasm. PMID- 17917821 TI - Comparison of three instrumental methods for the objective evaluation of radiotherapy induced erythema in breast cancer patients and a study of the effect of skin lotions. AB - A non-blinded three armed study of the effect of Aloe vera, Essex and no lotion on erythema was performed. The erythema is an effect of radiotherapy treatment in breast cancer patients. The study required testing of objective methods for measuring the erythema. The chosen experimental methods were Near Infrared Spectroscopy, Laser Doppler Imaging and Digital Colour Photography. The experimental setup was made in such a way that in parallel with testing the effect of the lotions there was also a test of the sensitivity of the instruments. Fifty women were selected consecutively to participate in the study. They were all subjected to treatment with high-energy electrons (9-20 MeV) after mastectomy, 2 Gy/day to a total dose of 50 Gy. Measurements were performed before the start of radiotherapy and thereafter once a week during the course of treatment. Aloe vera and Essex lotion were applied twice every radiation day in selected sites. The increase in skin redness could be monitored with all techniques with a detection limit of 8 Gy for Digital Colour Photography and Near Infrared Spectroscopy and 18 Gy for Laser Doppler Imaging. In clinical practice our recommendation is to use Digital Colour Photography. No significant median differences were observed between the pairs no lotion-Essex, no lotion-Aloe vera and Essex-Aloe vera for any of the techniques tested. PMID- 17917822 TI - Breast cancer patients' narratives about positive and negative communication experiences. AB - Health staff-patient communication is increasingly considered an important issue in cancer research. However, questionnaires addressing satisfaction with communication limit the issues patients can raise, do not address the context of communication and often show a strong positive skew in responses. Thus, qualitative studies of communication are also needed. Fifteen breast cancer patients were interviewed 3 months after finishing adjuvant treatment. They were asked to tell a 10 minute narrative and recall five experiences from treatment. Themes were extracted using categories derived from previous research while at the same time being sensitive to new elaborations and categories. The participants reported both positive and negative communication-related experiences from a wide range of treatment situations. Two major themes emerged: Information giving as professional care-giving and meeting emotional needs. The analysis suggests that appropriate information giving may have several functions, such as re-establishing the patient's future and reducing worst-case fantasies. Meeting emotional needs was seldom reported as directly talking about negative emotions, but rather through a variety of health staff behaviours. Also, the analysis points to problems in expecting or even pressurizing patients to feel and display negative emotions. The results highlight that meeting medical and emotional needs of patients may be closely intertwined in concrete treatment situations. PMID- 17917824 TI - Analysis of the dose calculation accuracy for IMRT in lung: a 2D approach. AB - The purpose of this study was to compare the dosimetric accuracy of IMRT plans for targets in lung with the accuracy of standard uniform-intensity conformal radiotherapy for different dose calculation algorithms. Tests were performed utilizing a special phantom manufactured from cork and polystyrene in order to quantify the uncertainty of two commercial TPS for IMRT in the lung. Ionization and film measurements were performed at various measuring points/planes. Additionally, single-beam and uniform-intensity multiple-beam tests were performed, in order to investigate deviations due to other characteristics of IMRT. Helax-TMS V6.1(A) was tested for 6, 10 and 25 MV and BrainSCAN 5.2 for 6 MV photon beams, respectively. Pencil beam (PB) with simple inhomogeneity correction and 'collapsed cone' (CC) algorithms were applied for dose calculations. However, the latter was not incorporated during optimization hence only post-optimization recalculation was tested. Two-dimensional dose distributions were evaluated applying the gamma index concept. Conformal plans showed the same accuracy as IMRT plans. Ionization chamber measurements detected deviations of up to 5% when a PB algorithm was used for IMRT dose calculations. Significant improvement (deviations approximately 2%) was observed when IMRT plans were recalculated with the CC algorithm, especially for the highest nominal energy. All gamma evaluations confirmed substantial improvement with the CC algorithm in 2D. While PB dose distributions showed most discrepancies in lower (<50%) and high (>90%) dose regions, the CC dose distributions deviated mainly in the high dose gradient (20-80%) region. The advantages of IMRT (conformity, intra-target dose control) should be counterbalanced with possible calculation inaccuracies for targets in the lung. Until no superior dose calculation algorithms are involved in the iterative optimization process it should be used with great care. When only PB algorithm with simple inhomogeneity correction is used, lower energy photon beams should be utilized. PMID- 17917823 TI - Clinical outcome in patients with prostate cancer treated with external beam radiotherapy and high dose-rate iridium 192 brachytherapy boost: a 6-year follow up. AB - To report the long-term results for treatment of localized carcinoma of the prostate using high dose rate (HDR) brachytherapy, conformal external beam radiotherapy (3D EBRT) and neo-adjuvant hormonal therapy (TAB). From 1998 through 1999, 154 patients with localized prostate cancer were entered in the trial. Biologically no evidence of disease (bNED) was defined at PSA levels < 2 microg/l. In order to compare the results of this treatment with other treatment modalities, the patient's pre-treatment data were used to calculate the estimated 5-year PSA relapse free survival using Kattan's nomograms for radical prostatectomy (RP) and 3D EBRT. After 6 years of follow-up, 129 patients remain alive. The actual 5-year relapse-free survival is 84%. None of the patients demonstrated clinical signs of local recurrence. The median PSA at follow-up among the relapse-free patients was 0.05 microg/l. Among the 80 patients who presented with clinical stage T3 tumours, 55 (68%) were relapse-free. The expected 5-year relapse-free survival using nomograms for RP and 3D EBRT was 54% and 70%, respectively. Late rectal toxicity RTOG grade 3 occurred in 1% of the patients. Late urinary tract toxicity RTOG grade 3 developed in 4% of the patients. Combined treatment, utilizing HDR, 3D EBRT and TAB, produces good clinical results. Rectal toxicity is acceptable. Urinary tract toxicity, most likely can be explained by the fact that during the first years of this treatment, no effort was made to localize the urethra, which was assumed to be in the middle of the prostate. PMID- 17917825 TI - Fractalkine: an important candidate for directing periglomerular leukocyte accumulation in irradiated mouse kidneys. AB - Radiation-induced impairment of renal function is preceded by capillary endothelial cell damage, which initiates a cascade of inflammatory and thrombotic events. Accumulation of leukocytes in the irradiated kidney, especially in areas surrounding the glomeruli, has been clearly demonstrated. The chemokine fractalkine has recently been identified as a key mediator of leukocyte adhesion that functions without the requirement of integrins or selectin-mediated rolling. In this study we investigate the possible involvement of fractalkine in the inflammatory response of the irradiated kidney. Mouse kidneys were irradiated with single doses of 16 or 0 Gy, and protein and mRNA levels of fractalkine and PECAM-1 were examined after 10 to 40 weeks. These changes were correlated with the progressive increase and distribution of leukocytes in the irradiated kidneys. Increased fractalkine immunoreactivity was seen at glomerular sites 30 to 40 weeks after irradiation. This fractalkine expression was strongly associated with the presence of leukocytes surrounding the Bowman's capsule of the same glomeruli. No significant changes in mRNA levels of fractalkine were seen in whole kidney extracts after irradiation, but expression levels were not determined for isolated glomeruli. PECAM-1 protein levels did not change with time after irradiation, although a significant decrease in mRNA expression was seen at 10 weeks. This study is the first demonstration of increased fractalkine after irradiation and the results suggest that fractalkine may be an important mechanism of leukocyte trafficking in the development of a radiation induced inflammatory response. PMID- 17917826 TI - Treatment schedule is of importance when gefitinib is combined with irradiation of glioma and endothelial cells in vitro. AB - Amplified epidermal growth factor receptor (EGFR) signaling is supposed to contribute to clinical radiation resistance of glioblastoma multiforme (GBM). Therefore, inhibition of EGFR signaling pathways by the selective EGFR tyrosine kinase inhibitor, gefitinib (ZD1839, Iressa), may increase the therapeutic effects of radiotherapy. The effects of different schedules for administration of gefitinib on sensitivity to irradiation of the human glioma cell lines (251MG and SF-767), a rat glioma cell line (BT4C), and an immortalized rat brain endothelial cell line (RBE4) is reported. Differences in effects of the combined treatment on cell toxicity were determined by a fluorometric cytotoxicity assay, and nuclear DNA fragmentation was used for quantification of apoptosis. Pre-administration with gefitinib for 30 min prior to irradiation followed by continuous incubation with gefitinib significantly increased the cytotoxicity of SF-767, BT4C, and RBE4 cells. However, the human glioma cell line 251MG was protected against radiation induced damage by this treatment schedule, at lower concentrations of gefitinib. Pre-administration with gefitinib for 24 h prior to irradiation without following incubation with gefitinib increased the cytotoxicity of SF-767 and BT4C cells. Post-irradiation treatment with gefitinib significantly increased the cytotoxicity in all cell lines except for 251MG. We demonstrated heterogeneity in the cytotoxic effects of gefitinib between cell lines. Response to gefitinib might be due to other mechanisms than through the EGF receptor as some of the cell lines showed sensitivity to gefitinib despite no or low expression of EGFR. This study also demonstrates the importance of timing of gefitinib administration when this agent is combined with irradiation. PMID- 17917827 TI - Immunization with truncated sequence of Telomerase Reverse Transcriptase induces a specific antitumor response in vivo. AB - To select the MHC-I-binding epitope-rich sequence of mice telomerase reverse transcriptase (mTERT) and study the antitumor immune response induced by truncated TERT through mRNA-transfected dendritic cells (DCs) immunization in mice. The MHC-I-binding epitopes of TERT were predicted using bioinformatics software. The selected sequence of TERT (Truncated mTERT, TERT(t), mTERT cDNA 1776 bp-2942 bp encoding 584 aa-969 aa) was cloned from B16 mouse melanoma cells and inserted into pBluescriptIIKS(+) plasmid downstream of the T7 promoter. TERT(t) RNA was prepared through in vitro transcription. Bone marrow-derived DCs were electroporated with TERT(t) RNA and used to immunize syngeneic naive mice. The quantity and cytotoxic activity of TERT-specific cytotoxic T lymphocytes (CTLs) in mice spleen were evaluated using IFN-gamma enzyme-linked immunospot (ELISPOT) and Lactate dehydrogenase release assay. The immunoprophylactic effects against TERT positive tumor induced by TERT(t) RNA transfected DC in vivo were evaluated through an immunized-challenged mouse model. TERT(t) was cloned and in vitro transcribed into TERT(t) mRNA. As shown in FCM analysis, the efficiency of DC electroporation is 35.1% (29.7-41.2%). After electroporation, a subtle increase of costimulator and MHC-II molecules were expressed on the cell surface. Immunization of TERT(t) mRNA transfected DCs induced IFN-gamma-secreting CTLs which manifested specific cytotoxic activity against TERT-positive target cells. In a cancer mouse model, vaccination of TERT(t) mRNA-transfected DCs suppressed the growth of TERT positive tumors (p=0.001) and prolong the survival time of tumor-bearing animals (p=0.029). TERT(t) evokes an antitumor immune response in vivo which is targeted to TERT. TERT(t) can be used as an antigeneic sequence to produce anti-TERT tumor vaccine. PMID- 17917828 TI - The number of analyzed lymph nodes - a prognostic factor in colorectal cancer. AB - The prognostic significance of the number of lymph nodes examined in surgical specimen of colorectal cancer was determined. One thousand and twenty five patients with colorectal cancer stage II and III were included in the study. These patients underwent surgery from 1991 to 1997 and were enrolled in clinical trials to evaluate the efficacy of adjuvant 5-fluorouracil (5FU) based chemotherapy. The median number of examined lymph nodes was five. Only 13% of the patients had > or = 12 lymph nodes analyzed. The number of examined lymph nodes was an independent prognostic factor for overall survival in the entire group of patients with stage II and III colorectal cancer (p=0.009). Patients with a higher number of lymph nodes examined had a longer overall survival. In stage III colorectal cancer the ratio of the number of metastatic lymph nodes to the number of examined lymph nodes (lymph node ratio, LNR) was an independent prognostic factor for overall survival. A decreasing LNR was correlated with a longer overall survival (p<0.0001). Increasing age was associated with a reduction of lymph node harvest (p=0.04). Patients with rectal cancer treated with preoperative radiotherapy had a lower number of lymph nodes analyzed compared with non-radiated (p<0.001). The number of examined lymph nodes in the surgical specimen is an independent prognostic factor for overall survival in colorectal cancer. The LNR is an independent prognostic factor in stage III colorectal cancer. PMID- 17917830 TI - Clinical utility of vascular endothelial growth factor in diagnosing malignant pleural effusions. AB - While the early diagnosis of cancer has been fully respected, it is still however often difficult for clinicians to confirm malignant pleural effusions (PE), which essentially indicate the end-stage cancer. It has now been demonstrated that vascular endothelial growth factor (VEGF) is a pivotal angiogenesis factor and associated with tumor growth and metastasis. The aim of this study was then to assess the diagnostic performance of VEGF in malignant PE. In this controlled and blinded prospective study, 113 consecutive patients with PE were recruited. For each eligible case, the VEGF levels of pleural fluid (PF) and serum were examined simultaneously using enzyme immunoassay. The reference standard for malignant PE was clinical evaluation and PF cytology with pleural biopsy, other examination and follow-up added as needed. According to the final diagnoses, 81 qualified cases were grouped as malignant (n=32) and benign (n=49) PE. For PF VEGF level, the mean in malignant group was higher than that in benign group (1358+/-1493 pg/mL vs. 422+/-317 pg/mL, p=0.001). As did for serum VEGF level (650+/-533 pg/mL vs. 137+/-189 pg/mL, p<0.001). Using receiver operating characteristic analysis, the determined diagnostic cut-off points of VEGF levels of PF and serum for malignant PE were 959.25 pg/mL and 212.36 pg/mL, with sensitivities of 47%, 69% and specificities of 96%, 88%, respectively. For cascade connection and parallel operation of PF VEGF and serum VEGF, the sensitivities were 34%, 81% at specificities of 98%, 86%, respectively. These findings suggest that VEGF could be used in diagnosing malignant PE as a useful adjunct of conventional algorithm. Different VEGF test strategies, including test on PF, serum and both, may be selected according to practical needs. PMID- 17917829 TI - Secondary treatment and predictive factors for second-line chemotherapy after first-line oxaliplatin-based therapy in metastatic colorectal cancer. AB - Two consecutive studies have evaluated the efficacy of oxaliplatin combined with the Nordic bolus schedule of 5-fluorouracil and folinic acid as first-line treatment in metastatic non-resectable colorectal cancer. One hundred and twelve patients were followed after end of first-line treatment and any secondary therapy registered. Fifty-three patients (47%) did not receive second-line irinotecan-based chemotherapy. The main reason was too poor performance status (59%). These patients had a median survival of only 1.7 months after progression of first-line therapy. The best predictive factors at start of first-line chemotherapy for receiving later second-line chemotherapy were performance status and alkaline phosphatase level. Fifty-nine patients (53%) received irinotecan based second-line therapy. Four (7%) patients had a partial response, and 28 (52%) had stable disease. Median progression-free survival after second-line chemotherapy was 4.1 months and median survival 9.5 months. Median survival after first-line chemotherapy and secondary liver surgery was 34 months and five-year disease-free survival 8%. Survival among patients receiving both first- and second-line chemotherapy was 20.8 months, but only 8.9 months in patients not receiving second-line irinotecan-based chemotherapy. Poor performance status or elevated alkaline phosphatase level at start of first-line chemotherapy predicts whether second-line chemotherapy will be given or not. PMID- 17917831 TI - FDG uptake in a rectal malignant melanoma. PMID- 17917832 TI - Capecitabine and oxaliplatin (XELOX) is safe and effective in patients with advanced gastric cancer. PMID- 17917833 TI - Gliomatosis Peritonei presenting as rectovaginal septum mass following recurrent mature ovarian teratoma. PMID- 17917834 TI - Adipose tissue formation in response to basic fibroblast growth factor. AB - CONCLUSION: Formation of new fat cells may occur in injected adipose tissue in response to basic fibroblast growth factor (bFGF). OBJECTIVE: In an effort to prevent loss of fat volume and generation of additional adipose tissue after intracordal injection of autologous fat, the effects of injecting fat together with basic fibroblast growth factor bFGF were evaluated. MATERIALS AND METHODS: Models of bilateral recurrent laryngeal nerve paralysis were prepared using 12 beagle dogs. Autologous fat was injected into one vocal cord, and a mixture of autologous fat with bFGF, gelatin microspheres, and collagen sponge was injected into the other. Histologic reactions were examined over time. RESULTS: In vocal cords where autologous fat was injected together with bFGF, fusiform immature adipocytes were found in the injected fat at 8 weeks after injection. The volume of the injected fat was maintained almost completely even at 24 weeks after injection. Vocal cords where only autologous fat was injected showed a marked decrease in volume of injected fat over time. PMID- 17917835 TI - Effect of an enlarged endolymphatic duct on bone conduction threshold. AB - An enlarged endolymphatic duct and sac (EDS) that makes contact with the cerebrospinal fluid-dural interface plays an important role in the pathway of bone conduction and enhances bone conduction at lower frequencies. Objectives. We investigated whether the bone conduction threshold was improved when the EDS was enlarged. Subjects and methods. Twenty-three patients (46 ears) with large vestibular aqueducts underwent standard pure tone audiometry (PTA) and magnetic resonance imaging (MRI) to investigate the relation between the diameter of the endolymphatic duct (ED) and the air or bone conduction threshold. We also investigated the relation between the volume of the EDS and the air or bone conduction threshold. Results. All ears had a mixed type hearing loss. The air bone gaps were significantly larger at 250 and 500 Hz than at higher frequencies. The bone conduction thresholds were significantly lower at 250 Hz and 1000 Hz when the diameter of the ED was large, whereas there was no relation between the diameter of the ED and the air conduction threshold. In addition, there was no correlation between the volume of the EDS and air or bone conduction thresholds. PMID- 17917836 TI - Proteomic analysis of two head and neck cancer cell lines presenting different radiation sensitivity. AB - CONCLUSION: We found four proteins in the AMC-HN-9 cells that might perform important functions in radio-resistance. These results also suggest that this cell line may provide us with important information about cancer cells regarding the roles of reactive oxygen species (ROS and antioxidants) in irradiation. OBJECTIVE: Radiation susceptibility can be determined by the expression of some proteins. To identify such proteins involved in radiation susceptibility, we analyzed two cell lines with different radiation sensitivity. MATERIALS AND METHODS: The AMC-HN-3 and -9 cell lines established from head and neck cancer patients were employed. They were irradiated with 4 Gy and two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was carried out. Then matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) was performed to identify 20 proteins. Western blotting was used to confirm the significantly expressed proteins. RESULTS: After 2-D PAGE, five spots were differentially expressed in the AMC-HN-9. Heat shock protein 27 kDa (HSP27), peroxiredoxin (Prx) II, glutathione S-transferase pi (GSTP), and an unknown protein were detected through MALDI-TOF MS. By using Western blotting, remarkable increments of the band intensity of HSP27, GSTP, Prx II, and Prx IV were confirmed in the AMC-HN-9 cell line. PMID- 17917837 TI - Clinicopathologic significance and prognostic role of cyclin E and cyclin A expression in laryngeal epithelial lesions. AB - CONCLUSIONS: The determination of cyclin A expression might be helpful in the identification of laryngeal squamous cell cancer (LSCC) patients with increased risk of metastases. The results suggest that cyclin A may be a more informative marker for cell proliferation than Ki-67. Abnormalities of cyclin E and cyclin A may play an important role in LSCC development and progression; however, the expression of cyclin E does not seem to have prognostic significance. OBJECTIVE: The aim of the study was to elucidate a possible association between cyclin E and cyclin A expression and clinicopathologic factors and their potential role as prognostic markers for patients with laryngeal epithelial lesions. MATERIALS AND METHODS: Expression of cyclins E and A, and Ki-67 was examined immunohistochemically in a formalin-fixed, paraffin-embedded series of 46 LSCC; 23 epithelial dysplasias (ED); and 21 normal mucosae (NM). RESULTS: The mean labeling indices (LIs) for cyclin E in LSCC, ED, and NM were 10.6%, 4.9%, and 0%, and for cyclin A 27.2%, 17.5%, and 7%, respectively. In LSCC, a statistically significant correlation was found between enhanced cyclin A expression and a higher incidence of locoregional lymph node metastasis (p0.01). The enhanced expression of cyclin A was linked with cell proliferation in LSCC, ED, and NM. No association was observed between cyclin E and A and other clinicopathologic parameters or applied treatments. The prognostic significance of cyclin E, cyclin A, and Ki-67 in determining overall survival time showed no statistically significant differences. PMID- 17917838 TI - Susceptibility to impulse noise trauma in different species: guinea pig, rat and mouse. AB - CONCLUSION: The results indicate that susceptibility to impulse noise differs in the three species. OBJECTIVE: To test the hypothesis that there is different susceptibility to impulse noise trauma in three species: the guinea pig, rat and mouse. MATERIALS AND METHODS: Three groups of animals were exposed to the same impulse noise and the effect on cochlear pathology was examined using auditory brainstem response thresholds. RESULTS: It was found that the rat and mouse are more sensitive to impulse noise than the guinea pig. PMID- 17917839 TI - The pharmacokinetic profiles of dexamethasone and methylprednisolone concentration in perilymph and plasma following systemic and local administration. AB - CONCLUSION: Both methylprednisolone (MTH) and dexamethasone (DEX) could successfully and effectively penetrate the round window membrane (RWM) into perilymph. RWM topical application and otocyst infusion with MTH and DEX result in high perilymph drug concentrations and low plasma levels. An intratympanic administration schedule for DEX or MTH could be carried out twice daily. OBJECTIVE: To explore the pharmacokinetics of DEX and MTH in the inner ear fluids and plasma following systemic and local administration. MATERIALS AND METHODS: Three routes of administration of steroids were used in guinea pigs: intracardial injection, otocyst infusion and RWM topical application by granule gelfoam soaked with steroids. Samples of blood or perilymph of the scala tympani were collected at 1-6 h after administration. High-performance liquid chromatography was used to assay concentrations of steroids. RESULTS: Both the topical application and infusion administration resulted in a significantly higher concentration of steroids in perilymph than intracardial injection. The level of steroids in the perilymph reached a peak at 1-2 h after dosing, and this was maintained at a relatively high level for several hours. The intracardial injection with steroids yielded very low perilymph levels at all sampling times after administration. PMID- 17917840 TI - Evaluation of combined medical and surgical treatment in nasal polyposis - III. Correlation between symptoms and CT scores before and after surgery for nasal polyposis. AB - CONCLUSION: Computed tomography (CT) in nasal polyposis (NP) patients has three functions before any treatment. CT provides objective evidence of the disease and precise topography of the disease, and is an indicator of the disease severity. After functional endoscopic sinus surgery (FESS), CT is an indicator of the residual disease severity and permits detection of asymptomatic mucoceles. OBJECTIVE: NP affects nearly 4% of the population. CT has become the examination of choice for the exploration of NP. FESS is accepted for NP treatment in the setting of failure of medical management. The aim of this study was to find out whether any correlation exists between symptom severity and CT scan score before and after FESS. PATIENTS AND METHODS: A total of 114 CT scans were performed in NP patients without contrast medium before and after FESS (mean follow-up 5 years), and were scored according to the Lund-MacKay system. RESULTS: Lund-MacKay scores before treatment ranged from 8 to 24. There was a correlation between symptom and CT scores before any treatment. Postoperative Lund-MacKay scores ranged from 0 to 24. There was a correlation between symptom and CT scores after surgery. There was no correlation between postoperative symptom and baseline CT scores. Eleven asymptomatic mucoceles were found. PMID- 17917841 TI - Nasal complaints and signs of disease in farmers--a methodological study. AB - CONCLUSION: The methods used in this study are suitable for field studies that involve examinations of groups of workers. For individual examinations, there is no gold standard method that can discriminate work-related discomfort from other causes of rhinitis. OBJECTIVES: Studies of the effects of occupation on farmers' health have mainly focused on lower airways; few studies have examined effects on upper airways. This study investigated nasal functions in three groups of farmers (swine, milk and grain producers) and a control group using different methods, suitable for field studies. SUBJECTS AND METHODS: Health-related complaints were examined and several functional tests, such as expirogram, olfactory threshold test, acoustic rhinometry, nasal lavage with biomarkers of inflammation (eosinophilic cationic proteins (ECP), myeloperoxidase (MPO), tryptase, albumin) and allergy tests were performed. The different tests were correlated to nasal complaints and to each other. RESULTS: Nasal blockage complaints were more common among farmers; overall, nasal polyps were more frequent in grain producers. Objective parameters showed more pronounced mucosal swelling in farmers and higher concentrations of ECP in nasal lavage compared with controls. Lung function, olfactory threshold, atopy frequency and allergen-specific IgE to the storage mite Lepidoglyphus destructor did not differ between farmers and controls. Mucosal swelling measured with acoustic rhinometry was more pronounced in subjects with nasal complaints, hypersensitivity, nasal polyps and symptoms from lower airways. There was a correlation between biomarkers in nasal lavage (MPO, albumin and ECP). PMID- 17917844 TI - Abstracts of the 5th International Meeting on Intensive Cardiac Care, October 14 16, 2007, Tel Aviv, Israel. PMID- 17917847 TI - We have a problem: why have ALS trials been negative? PMID- 17917842 TI - Preoperative vestibular ablation with gentamicin and vestibular 'prehab' enhance postoperative recovery after surgery for pontine angle tumours--first report. AB - CONCLUSIONS: Preoperative gentamicin in combination with vestibular 'prehab' offers a possibility to reduce postoperative malaise and speed up recovery and may be used for patients undergoing such surgery when there is remaining vestibular function. OBJECTIVES: Removal of pontine angle tumours in a patient with remaining vestibular function causes symptoms of acute vestibular loss. A simultaneous cerebellar lesion can cause a combined vestibule-cerebellar lesion. PATIENTS AND METHODS: Twelve patients with pontine angle tumours but with near normal vestibular function were treated with intratympanic gentamicin in combination with vestibular 'prehab' to achieve preoperative vestibular ablation and compensation. After work-up patients started with a home-based vestibular training programme for 14 days. They then received a total of 1.2 ml of 30 mg/ml buffered gentamicin in four intratympanic installations over 2 days. They continued training and returned 6-16 weeks later. All patients were tested with calorics, vestibular video-impulse testing of all six canals, VEMP, subjective visual vertical and horizontal, posturography and pure tone and speech audiometry. RESULTS: There was a loss of caloric reactions and loss of impulses. In two patients the hearing deteriorated and in one hearing improved. All subjects were vestibulary compensated before surgery and no patient complained of dizziness or vertigo after surgery. PMID- 17917848 TI - Survey of ALS-associated factors potentially promoting Ca2+ overload of motor neurons. AB - The deleterious consequences of Ca(2+) overload are thought to be a probable cause of motoneuronal death in ALS, although the overloading mechanism is currently unclear. In this paper some ALS-linked factors are analysed with regard to their influence on Ca(2+ )influx into neurons. Intensive cortex activity can render motor neurons susceptible to stimulation of calcium-permeable glutamate NMDA-receptors; increase in CSF concentrations of glutamate, glycine, and norepinephrine supposedly can intensify these receptors' activity. Elevated CSF levels of GABA and reduced levels of serotonin can promote Ca(2+ )influx through glutamate AMPA-receptors and voltage-gated channels of L-, N-, and P-type. Additionally, brain ischaemia can contribute to Ca(2+ )overload of motor neurons. Thus, ALS is characterized by the unique combination of factors potentially able to promote the overload of motor neurons with calcium. PMID- 17917849 TI - Distinctions between the dementia in amyotrophic lateral sclerosis with frontotemporal dementia and the dementia of Alzheimer's disease. AB - The clinical entity of Amyotrophic Lateral Sclerosis with Frontotemporal Dementia (ALS-FTD) has only recently been recognized as an important neurodegenerative disease. As in isolated FTD, the behavioral and personality changes in ALS-FTD might be more characteristic than its cognitive changes. We aimed to characterize the behavioral and cognitive deficits in ALS-FTD, and contrast this profile with that of the most common form of dementia, AD, to assist ALS clinicians in recognizing the syndrome early in its course. Specifically, we hypothesized that a modified version of the Frontal Behavioral Inventory (FBI-mod), a brief questionnaire self-administered by a caregiver, along with just a few cognitive tests, would be clinically useful in distinguishing the dementia in ALS-FTD from the dementia of AD. We administered a battery of neuropsychological tests to 15 patients who met established criteria for Amyotrophic Lateral Sclerosis with Frontotemporal Dementia and to 30 patients who met established criteria for probable Alzheimer's disease. The FBI-mod was completed by caregivers. We found that the FBI-mod, age-corrected Z scores for the Mini-Mental State Examination (MMSE), a test of delayed recall, and a word fluency measure together discriminated between ALS-FTD and AD. ALS-FTD was characterized by more abnormal FBI scores and poor word fluency, in the presence of relatively normal overall cognitive status (MMSE) and/or delayed recall. PMID- 17917850 TI - Olfactory ensheathing glia injections in Beijing: misleading patients with ALS. AB - Different forms of cell transplantation therapy are being tested in models of ALS. While the approach offers hope to patients with ALS, much still needs to be learned in the laboratory before it is ready for human trials. Nevertheless, clinics across the world offer various types of open label cell transplantation therapy for high fees. We report a woman who received an injection into each frontal lobe in Beijing, China. Her ALS progressed at a more rapid rate after the procedure and she suffered disabling side-effects. Clinics that give experimental and potentially harmful treatments outside the construct of well-designed clinical trials put patients at risk and do a disservice to the ALS community. PMID- 17917852 TI - Intraperitoneal administration of CDP-choline and its cholinergic and pyrimidinergic metabolites induce hyperglycemia in rats: involvement of the sympathoadrenal system. AB - CDP-choline is an endogenous metabolite in phosphatidylcholine biosynthesis. Exogenous administration of CDP-choline has been shown to affect brain metabolism and to exhibit neuroprotective actions. On the other hand, little is known regarding its peripheral actions. Intraperitoneal administration of CDP-choline (200-600 micromol/kg) induced a dose- and time-dependent hyperglycemia in rats. Hyperglycemic response to CDP-choline was associated with several-fold elevations in serum concentrations of CDP-choline and its metabolites. Intraperitoneal administration of phosphocholine, choline, cytidine, cytidine monophosphate, cytidine diphosphate, cytidine triphosphate, uridine, uridine monophosphate, uridine diphosphate and uridine triphosphate also produced significant hyperglycemia. Pretreatment with atropine methyl nitrate failed to alter the hyperglycemic responses to CDP-choline and its metabolites whereas hexamethonium, the ganglionic nicotinic receptor antagonist which blocks nicotinic cholinergic neurotransmission at the autonomic ganglionic level, blocked completely the hyperglycemia induced by CDP-choline, phosphocholine and choline, and attenuated the hyperglycemic response to cytidine monophosphate and cytidine. Increased blood glucose following CDP-choline, phosphocholine and choline was accompanied by elevated plasma catecholamine concentrations. Hyperglycemia elicited by CDP choline and its metabolites was entirely blocked either by pretreatment with a nonselective -adrenoceptor antagonist phentolamine or by the 2-adrenoceptor antagonist, yohimbine. Hyperglycemic responses to CDP-choline, choline, cytidine monophosphate and cytidine were not affected by chemical sympathectomy, but were prevented by bilateral adrenalectomy. Phosphocholine-induced hyperglycemia was attenuated by bilateral adrenalectomy or by chemical sympathectomy. These data show that CDP-choline and its metabolites induce hyperglycemia which is mediated by activation of ganglionic nicotinic receptors and stimulation of catecholamine release that subsequently activates 2-adrenoceptors. PMID- 17917853 TI - Peptides other than the neurotrophins that can be cleaved from proneurotrophins: a neglected story. AB - The members of the family of neurotrophic factors known as neurotrophins, NGF, BDNF, NT-3 and NT4/5 are known to be cleaved intracellularly from immature precursors, the proneurotrophins. NGF and the other neurotrophins regulate neurite outgrowth and neuronal survival during development via binding to Trk receptor tyrosine kinases and the p75 neurotrophin receptor. Surprisingly, the proneurotrophins were shown to be also biologically active ligands. ProNGF and proBDNF induce neuronal apoptosis via binding to a complex of p75 and sortilin. Therefore, life and death seems to be a delicate interplay between 'cleavage' or 'not cleavage' of the proneurotrophins. However, there is a third aspect to this story. In general, peptide-hormone precursors are known to give rise to several biologically active peptides from one precursor molecule. The paradox with the proneurotrophins is that although they have several additional potential cleavage sites that would necessarily give rise to other peptides besides the neurotrophins and thus new members in the neurotrophin family, this aspect has been largely neglected. This article aims to review evidence for biologically active peptides other than the NGF and NT-3 that can be generated from the proNGF and proNT-3. PMID- 17917854 TI - A microaliquoting technique for precise histological annotation and optimization of cell content in frozen tissue specimens. AB - Knowledge of the exact cell content of frozen tissue samples is of growing importance in genomic research. We developed a microaliquoting technique to measure and optimize the cell composition of frozen tumor specimens for molecular studies. Frozen samples of 31 mesothelioma cases were cut in alternating thin and thick sections. Thin sections were stained and evaluated visually. Thick sections, i.e., microaliquots, were annotated using bordering stained sections. A range of cellular heterogeneity was observed among and within samples. Precise annotation of samples was obtained by integration and compared to conventional single face and "front and back"' section estimates of cell content. Front and back estimates were more highly correlated with block annotation by microaliquoting than were single face estimates. Both methods yielded discrepant estimates, however, and for some studies may not adequately account for the heterogeneity of mesothelioma or other malignancies with variable cellular composition. High yield and quality RNA was extracted from precision annotated, tumor-enriched subsamples prepared by combining individual microaliquots with the highest tumor cellularity estimates. Microaliquoting provides accurate cell content annotation and permits genomic analysis of enriched subpopulations of cells without fixation or amplification. PMID- 17917855 TI - Effects of fixation on routine, special and immunohistochemical stains: introduction to a symposium for the Biological Stain Commission. PMID- 17917856 TI - The role of glycine and prolines in connective tissue fiber staining with hydrogen bonding dyes. AB - Extensive hydrogen bonding of dyes to connective tissue fibers is made possible by the high content of the amino acids proline and glycine in elastin and collagens. Proline confers an extended polypeptide structure and glycine is the only amino acid whose specific side group, -H, is so small that it forms no obstacle to hydrogen bonding between the peptide group and external molecules. Thus, a high proportion of the peptide groups in fibrous proteins are directly accessible to hydrogen bonding groups dye molecules. PMID- 17917863 TI - Hypertension and cardiovascular disease: is arterial stiffness the heart of the matter? PMID- 17917864 TI - ESH Hypertension Excellence Centres: a new strategy to combat an old foe. PMID- 17917865 TI - European Society of Hypertension Excellence Centres. PMID- 17917866 TI - The Test of Variables of Attention (TOVA): internal consistency (Q1 vs. Q2 and Q3 vs. Q4) in children with attention deficit/hyperactivity disorder (ADHD). AB - The internal consistency of the Test of Variables of Attention (TOVA) was examined in a cohort of 6- to 12-year-old children (N = 63) strictly diagnosed with ADHD. The internal consistency of errors of omission (OMM), errors of commission (COM), response time (RT), and response time variability (RTV) of different test conditions (stimulus infrequent condition [Q1 vs. Q2] and stimulus frequent condition [Q3 vs. Q4]) was assessed via correlation analyses. All TOVA index scores under investigation assessing its internal consistency exhibited statistically significant correlations. All correlations fell in the moderate high range. PMID- 17917867 TI - Unusual complications of heroin abuse: transverse myelitis, rhabdomyolysis, compartment syndrome, and ARF. AB - INTRODUCTION: Heroin overdose can cause various rare neurological complications like spongiform leukoencephalopathy, seizures, stroke, toxic amblyopia, transverse myelopathy, mononeuropathy, plexopathy, acute inflammatory demyelinating polyradiculoneuropathy, rhabdomyolysis, compartment syndrome, fibrosing myopathy, and acute bacterial myopathy. We report here the simultaneous presentation of multiple complications of heroin toxicity. CASE REPORT: A young heroin addict was found unarousable lying in the lotus posture. Examination showed quadriplegia and left leg gangrene. He subsequently developed heroin induced transverse myelitis, rhabdomyolysis, left leg compartment syndrome, and myoglobin-induced acute renal failure. DISCUSSION: This case leads us to consider a common linked or systemic mechanism of injury rather than a local mechanism when multiple simultaneous organ failure occurs complicating heroin abuse. PMID- 17917868 TI - Neonatal thrombocytosis following G-CSF treatment. AB - Thrombocytosis is defined as an elevation of the platelet count to more than 500,000/mm(3). Primary thrombocytosis rarely occurs in the pediatric age group and is usually caused by a clonal bone marrow disorder. The more common phenomenon is secondary thrombocytosis which is a reactive process. Table 1 lists the main causes of secondary thrombocytosis. Complications of severe thrombocytosis include bleeding and thromboses. Unless additional risk factors are present, secondary thrombocytosis is not associated with a significant risk of thromboembolic events, regardless of the degree of elevation of the platelet count. The aim of this case report is to add a new possible cause of neonatal thrombocytosis. PMID- 17917870 TI - Stationary phase mutagenesis in B. subtilis: a paradigm to study genetic diversity programs in cells under stress. AB - One of the experimental platforms to study programs increasing genetic diversity in cells under stressful or nondividing conditions is adaptive mutagenesis, also called stationary phase mutagenesis or stress-induced mutagenesis. In some model systems, there is evidence that mutagenesis occurs in genes that are actively transcribed. Some of those genes may be actively transcribed as a result of environmental stress giving the appearance of directed mutation. That is, cells under conditions of starvation or other stresses accumulate mutations in transcribed genes, including those transcribed because of the selective pressure. An important question concerns how, within the context of stochastic processes, a cell biases mutation to genes under selection pressure? Because the mechanisms underlying DNA transactions in prokaryotic cells are well conserved among the three domains of life, these studies are likely to apply to the examination of genetic programs in eukaryotes. In eukaryotes, increasing genetic diversity in differentiated cells has been implicated in neoplasia and cell aging. Historically, Escherichia coli has been the paradigm used to discern the cellular processes driving the generation of adaptive mutations; however, examining adaptive mutation in Bacillus subtilis has contributed new insights. One noteworthy contribution is that the B. subtilis' ability to accumulate chromosomal mutations under conditions of starvation is influenced by cell differentiation and transcriptional derepression, as well as by proteins homologous to transcription and repair factors. Here we revise and discuss concepts pertaining to genetic programs that increase diversity in B. subtilis cells under nutritional stress. PMID- 17917871 TI - Controlling mutation: intervening in evolution as a therapeutic strategy. AB - Mutation is the driving force behind many processes linked to human disease, including cancer, aging, and the evolution of drug resistance. Mutations have traditionally been considered the inevitable consequence of replicating large genomes with polymerases of finite fidelity. Observations over the past several decades, however, have led to a new perspective on the process of mutagenesis. It has become clear that, under some circumstances, mutagenesis is a regulated process that requires the induction of pro-mutagenic enzymes and that, at least in bacteria, this induction may facilitate evolution. Herein, we review what is known about induced mutagenesis in bacteria as well as evidence that it contributes to the evolution of antibiotic resistance. Finally, we discuss the possibility that components of induced mutation pathways might be targeted for inhibition as a novel therapeutic strategy to prevent the evolution of antibiotic resistance. PMID- 17917869 TI - Genetic constraints on protein evolution. AB - Evolution requires the generation and optimization of new traits ("adaptation") and involves the selection of mutations that improve cellular function. These mutations were assumed to arise by selection of neutral mutations present at all times in the population. Here we review recent evidence that indicates that deleterious mutations are more frequent in the population than previously recognized and that these mutations play a significant role in protein evolution through continuous positive selection. Positively selected mutations include adaptive mutations, i.e. mutations that directly affect enzymatic function, and compensatory mutations, which suppress the pleiotropic effects of adaptive mutations. Compensatory mutations are by far the most frequent of the two and would allow potentially adaptive but deleterious mutations to persist long enough in the population to be positively selected during episodes of adaptation. Compensatory mutations are, by definition, context-dependent and thus constrain the paths available for evolution. This provides a mechanistic basis for the examples of highly constrained evolutionary landscapes and parallel evolution reported in natural and experimental populations. The present review article describes these recent advances in the field of protein evolution and discusses their implications for understanding the genetic basis of disease and for protein engineering in vitro. PMID- 17917872 TI - The Hsp90 capacitor, developmental remodeling, and evolution: the robustness of gene networks and the curious evolvability of metamorphosis. AB - Genetic capacitors moderate expression of heritable variation and provide a novel mechanism for rapid evolution. The prototypic genetic capacitor, Hsp90, interfaces stress responses, developmental networks, trait thresholds and expression of wide-ranging morphological changes in Drosophila and other organisms. The Hsp90 capacitor hypothesis, that stress-sensitive storage and release of genetic variation through Hsp90 facilitates adaptive evolution in unpredictable environments, has been challenged by the belief that Hsp90-buffered variation is unconditionally deleterious. Here we review recent results supporting the Hsp90 capacitor hypothesis, highlighting the heritability, selectability, and potential evolvability of Hsp90-buffered traits. Despite a surprising bias toward morphological novelty and typically invariable quantitative traits, Hsp90-buffered changes are remarkably modular, and can be selected to high frequency independent of the expected negative side-effects or obvious correlated changes in other, unselected traits. Recent dissection of cryptic signal transduction variation involved in one Hsp90-buffered trait reveals potentially dozens of normally silent polymorphisms embedded in cell cycle, differentiation and growth control networks. Reduced function of Hsp90 substrates during environmental stress would destabilize robust developmental processes, relieve developmental constraints and plausibly enables genetic network remodeling by abundant cryptic alleles. We speculate that morphological transitions controlled by Hsp90 may fuel the incredible evolutionary lability of metazoan life-cycles. PMID- 17917875 TI - Adoptive immunotherapy of HCMV infection. AB - Human cytomegalovirus (HCMV) infection or reactivation is a frequent cause of morbidity and mortality in immunocompromised individuals such as transplant recipients. Primary HCMV infection or reactivation of HCMV from latency is mostly asymptomatic in immunocompetent individuals and is controlled by the host's cell mediated immune response. Healthy HCMV seropositive individuals develop high frequencies of HCMV-specific cytotoxic T lymphocytes (CTL) in the peripheral blood. Furthermore, a direct correlation between the recovery of HCMV-specific CTL responses and an improved outcome of HCMV disease could be demonstrated in immunocompromised patients. Deriving from these observations, the strategy of an adoptive transfer of HCMV-specific T cells has been developed. Protective immunity can be transferred successfully by the infusion of donor-derived HCMV specific CD8+ cytotoxic T-cell clones or cell lines. In addition, several studies have supported the importance of antiviral effector functions of Th cells in maintaining CTL responses after adoptive transfer and their capacity to produce antiviral cytokines. Until today, a broad variety of clinical protocols for HCMV specific immunotherapy has been published. These protocols vary regarding the isolation procedure, composition of cellular product, number of transferred cells and thus treatment efficacy. In this review, we aim to provide a comprehensive synopsis of the current standard of knowledge concerning cellular HCMV-specific immunotherapeutic approaches. PMID- 17917876 TI - Utilization of a test gradient enhances islet recovery from deceased donor pancreases. AB - BACKGROUND: Islet transplantation is a viable treatment alternative for a select group of patients with type 1 diabetes. However, variables unique to the donor pancreas, such as age, fibrosis and edema, can influence the number and purity of the isolated islets. Thus isolation of a sufficient number of islets for transplantation from the pancreas remains challenging because of the lack of methods enabling reproducible isolation. METHODS: Islets were isolated from 38 consecutive deceased donors using the semi-automated Ricordi method of islet isolation, and purified on a COBE 2991 cell processor using Ficoll-based continuous density gradients. Three different gradient protocols were used. These included a pre-defined gradient using different densities of Ficoll (1.100 g/mL and 1.077 g/mL) mixed with HBSS (group 1), a pre-defined gradient using single density Ficoll (1.100 g/mL) mixed with University of Wisconsin solution (UW) (group 2) and a variable gradient using single-density Ficoll (1.100 g/mL) with UW and densities selected based on the results of test gradients (group 3). RESULTS: Group 3 yielded a better recovery of islets (74%) than groups 1 (43%) or 2 (37%) (P=0.0144). Viability was significantly higher in groups 2 and 3 (P=0.0115). Purity was not significantly different among the groups. DISCUSSION: This method, using a simple test gradient, is a significant process improvement that can improve islet recovery without loss of viability or purity and increase the number of islet products suitable for transplantation. PMID- 17917877 TI - Low serum and serum-free culture of multipotential human adipose stem cells. AB - BACKGROUND: Adipose tissue provides an easily accessible and abundant source of putative stem cells for translational clinical research. Currently prevalent culture techniques include the use of FBS, a highly variable and undefined component, which brings with it the potential for adverse patient reactions. In an effort to eliminate the use of animal products in human adipose stem cell (ASC) cultures, we have developed two new culture methods, a very low human serum expansion medium and a completely serum-free medium. METHODS: Through serial testing, a highly enriched medium formulation was developed for use with and without the addition of 0.5% human serum, an amount easily obtainable from autologous blood draws. RESULTS: Very low-serum culture yielded population doubling times averaging 1.86 days in early passage, while the serum-free formulation was associated with less robust cell growth, with doubling times averaging 5.79 days. ASC in both conditions maintained its ability to differentiate into adipo-, chondro- and osteogenic lineages in vitro, despite lower expression of CD34 in early passage. Expression of ALDH, HLA, CD133, CD184, and CD31 was comparable with that seen in cells cultured in 10% FBS. DISCUSSION: These newly developed culture conditions provide a unique environment within which to study ASCs without the interference of animal serum, and allow for the rapid expansion of autologous ASCs in culture in an animal product-free environment for use in human clinical trials. PMID- 17917878 TI - Are UC blood transplants prone to developing leukemia? PMID- 17917873 TI - Stress-induced mutagenesis in bacteria. AB - Bacteria spend their lives buffeted by changing environmental conditions. To adapt to and survive these stresses, bacteria have global response systems that result in sweeping changes in gene expression and cellular metabolism. These responses are controlled by master regulators, which include: alternative sigma factors, such as RpoS and RpoH; small molecule effectors, such as ppGpp; gene repressors such as LexA; and, inorganic molecules, such as polyphosphate. The response pathways extensively overlap and are induced to various extents by the same environmental stresses. These stresses include nutritional deprivation, DNA damage, temperature shift, and exposure to antibiotics. All of these global stress responses include functions that can increase genetic variability. In particular, up-regulation and activation of error-prone DNA polymerases, down regulation of error-correcting enzymes, and movement of mobile genetic elements are common features of several stress responses. The result is that under a variety of stressful conditions, bacteria are induced for genetic change. This transient mutator state may be important for adaptive evolution. PMID- 17917879 TI - Selection of antisense oligonucleotides for reversal of multidrug resistance in breast carcinoma cells. AB - BACKGROUND: Multidrug resistance (MDR) is a major obstacle in cancer treatment. In the present study, six regions of the mdr1 gene associated with transcription control or translation initiation were selected as targets. Six antisense oligonucleotides (ASODN; AS1-AS6) complementary to the corresponding sequence of the mdr1 gene were synthesized to investigate whether or not blocking the transcription control sites with ASODN could reverse MDR and which ASODN had the best efficiency for reversing MDR in breast carcinoma cells. METHODS: Forty-eight hours after transfection, mdr1 mRNA and P-glycoprotein (Pgp) were determined by RT-PCR, flow cytometry and Rhodamine 123 (Rh123) retention assay. The chemosensitivity of the treated cells was evaluated by MTT assay. RESULTS: A significant reduction in expression of mdr1 mRNA and Pgp was found in four groups (AS1, AS3, AS5 and AS6), accompanying a dysfunction of Pgp. The lowest levels of mdr1 index and Pgp expression were observed in the AS6 group. MTT assay showed that a significant reduction of drug resistance was found in the four groups, especially in the AS6 group, which showed an 8.4-fold reduction in drug resistance for adriamycin and a 10.5-fold reduction in drug resistance for vinblastine. DISCUSSION: These data suggest that the MDR phenotype of breast carcinoma cells could be reversed by ASODN complementary to the transcription control site or translation initiation region. AS6, which is complementary to the translation initiation codon (ATG) of mdr1 cDNA, has the best reversal efficiency. PMID- 17917880 TI - Ab therapy of AML: native anti-CD33 Ab and drug conjugates. AB - MAb have become an important treatment modality in cancer therapy.Genetically engineered chimeric and humanized Ab have demonstrated activity against a variety of tumors. While the humanized anti-CD33MAb lintuzumab has only modest single agent activity against overt AML, it can eliminate minimal residual disease detectable by reverse transcription-PCR in acute promyelocytic leukemia. Targeted chemotherapy with the anti-CD33-calicheamicin construct gemtuzumab ozogamicin has produced remissions in patients with relapsed AML and appears promising when used in combination with standard chemotherapy in the treatment of newly diagnosed AML. PMID- 17917874 TI - Mutation as a stress response and the regulation of evolvability. AB - Our concept of a stable genome is evolving to one in which genomes are plastic and responsive to environmental changes. Growing evidence shows that a variety of environmental stresses induce genomic instability in bacteria, yeast, and human cancer cells, generating occasional fitter mutants and potentially accelerating adaptive evolution. The emerging molecular mechanisms of stress-induced mutagenesis vary but share telling common components that underscore two common themes. The first is the regulation of mutagenesis in time by cellular stress responses, which promote random mutations specifically when cells are poorly adapted to their environments, i.e., when they are stressed. A second theme is the possible restriction of random mutagenesis in genomic space, achieved via coupling of mutation-generating machinery to local events such as DNA-break repair or transcription. Such localization may minimize accumulation of deleterious mutations in the genomes of rare fitter mutants, and promote local concerted evolution. Although mutagenesis induced by stresses other than direct damage to DNA was previously controversial, evidence for the existence of various stress-induced mutagenesis programs is now overwhelming and widespread. Such mechanisms probably fuel evolution of microbial pathogenesis and antibiotic resistance, and tumor progression and chemotherapy resistance, all of which occur under stress, driven by mutations. The emerging commonalities in stress-induced mutation mechanisms provide hope for new therapeutic interventions for all of these processes. PMID- 17917881 TI - Comparative analysis of mesenchymal stromal cells from murine bone marrow and amniotic fluid. AB - BACKGROUND: Murine mesenchymal stromal cells (mMSC) are a good model for pre clinical investigations, and alternative sources of mMSC are subject to intensive experiments. In the present study, we obtained mMSCs from amniotic fluid (AF) and compared their characteristics with mMSCs from bone marrow (BM). METHODS: NMRI mice, 4-6 weeks old, were killed and AF and BM cells collected from those in the second week of pregnancy. MSC were achieved by adhesion to cell culture plastic. Isolated cells were assessed for clonogenic capacity, some surface markers (epitopes) and differentiation potential. RESULTS: We achieved AF mMSC more readily than BM mMSC. Differences concerning colony assay and some surface markers of mMSC derived from these two source cells were observed. Most strikingly, AF mMSC showed no adipogenic differentiation capacity, in contrast with BM mMSC. DISCUSSION: Our results show that mMSC from AF are an appropriate source for pre-clinical investigation. Furthermore, mMSC from different sources of mice vary in clonogenic capacity, surface markers and differentiation potential. PMID- 17917882 TI - Ethnically mismatched cord blood transplants in African Americans: the Saint Louis Cord Blood Bank experience. AB - BACKGROUND: For ethnic minority patients where a suitably matched BM or peripheral blood donor is frequently unavailable, cord blood offers an opportunity for hematopoietic stem cell transplantation. Focused recruitment of ethnic minorities for cord blood donation has been proposed as the preferred strategy to improve access for minority recipients to cord blood for transplantation. The aim of this study was to evaluate cord blood characteristics for Caucasian and African American donors and the success of ethnically mismatched UC blood transplantation in African American recipients. METHODS: Retrospective data analysis was performed comparing the characteristics of 556 cord blood units from African American and Caucasian donors. The outcomes of 18 African American ethnically mismatched transplant recipients were compared with a paired sample of 18 ethnically matched Caucasian recipients. RESULTS: The fraction of collected units meeting acceptability criteria from African Americans was significantly lower compared with Caucasians (P = <0.0001). Additionally, African Americans had a significantly lower post-processing total nucleated cell count (TNC) compared with Caucasians (P=0.007) but there were no other significant differences in conventionally measured product characteristics. In the transplant analysis, there was no difference in overall survival at 1 year (P=0.85) or time to neutrophil engraftment (P=0.92) between the two patient populations. DISCUSSION: At comparable levels of TNC dose and HLA matching, the use of ethnically mismatched UC blood units as a source for allogeneic unrelated transplant can result in successful transplant outcomes for African American patients. PMID- 17917883 TI - Proliferation, differentiation and characterization of osteoblasts from human BM mesenchymal cells. AB - BACKGROUND: The objective of this study was to isolate osteoprogenitor cells (OPC) from BM mesenchymal stromal cells (MSC) and test their capacity to proliferate and differentiate into osteoblasts. METHODS: Human MSC were separated on a Percoll gradient and cultured in DMEM supplemented with 15% human serum, and characterized by flow cytometric analyzes for CD34, CD13, CD90, CD105 and CD117. To induce differentiation, cultured cells were exposed to 10(-7) m dexamethasone (dexa) and/or 10(-3) m sodium beta-glycerophosphate (beta-GlyP) and 1,25 dihydroxyvitamin D3 (calcitriol) or 9-cis-retinoic acid (9-RA). RESULTS: alkaline phosphatase (AP) activity was detected in cells irrespective of the dexa and/or beta-GlyP treatment. Antigenic phenotypes of MSC were CD34- (more than 99%) and CD13+ CD90+ CD105+ CD117+ (c. 50%). The treatment induced extracellular calcium deposition and gene and protein expression of osteonectin (ON) and bone sialoprotein (BSP): beta-GlyP induced an increase (c. 2.2-fold) of the ON gene and dexa augmented (c. 2.7-fold) the gene expression of BSP II. Gene expression of BSP I reached a maximum at 3 weeks of combined treatment. Osteocalcin gene expression was induced only after additional treatment with calcitriol or 9-RA. Ultrastructural analysis revealed the secretory phenotype of OPC. DISCUSSION: Under appropriate treatment, MSC can give rise to OPC that have the capacity to differentiate into osteoblasts characterized by the expression of osteogenic markers, osteoblastic properties and stromal BM cells phenotypes. These cells may represent a promising material to be utilized in orthopedic cellular therapy. PMID- 17917884 TI - Transgene-enforced co-stimulation of CD4+ T cells leads to enhanced and sustained anti-tumor effector functioning. AB - Background The role of co-stimulation in CD4+ T cell activation by professional APC is well established, while less is known of the role co-stimulation plays when CD4+ T cells interact directly with tumor cells. Methods Through genetic engineering of human CD4+ T cells, we tested the hypothesis that integration of co-stimulatory signaling domains within a tumor-targeting chimeric Ag receptor (CAR), the IL-13Ralpha2-specific IL-13-zetakine (IL13zeta), would enhance CD4+ T cell mediated responses against tumors that fail to express ligands for co stimulatory receptors. Results Compared with CD3zeta-mediated activation alone, CD4+ effector T cells expressing the IL13-CD28-41BBzeta CAR exhibited augmented/sustained MAPK and AKT activity, up-regulated Th1 cytokine production, and enhanced cytolytic potency against tumor targets. Moreover, upon recursive stimulation with tumor, the IL13-CD28-41BBzeta+ cells retained/recycled their lytic function, whereas IL-13zeta+ CD4+ cells became anergic/exhausted. These in vitro observations correlated with enhanced in vivo control of established orthotopic CNS glioma xenografts in immunodeficient mice mediated by adoptively transferred ex vivo-expanded CD4+ T cells expressing the co-stimulatory CAR. Discussion Together these studies demonstrate the importance of integrating co stimulation with CD3zeta signaling events to activate fully CD4+ anti-tumor effector cells for sustained function in the tumor microenvironment. PMID- 17917885 TI - Activated beta-catenin induces myogenesis and inhibits adipogenesis in BM-derived mesenchymal stromal cells. AB - BACKGROUND: Mesenchymal stromal cells (MSC) have been thought to be attractive candidates for the treatment of degenerative muscle diseases. However, little is known about the molecular mechanisms governing the myogenic differentiation in MSC. As the Wnt signaling pathway has been associated with myogenesis in embryogenesis and post-natal muscle regeneration, we hypothesized that the Wnt signaling pathway may be involved in governing the myogenic differentiation in MSC. METHODS: Primary MSC were isolated from Sprague-Dawley rats and expanded in proliferation medium. The rMSC were transfected with a constitutively active hbeta-catenin (S37A) plasmid or control vector by Lipofectamine followed by G418 selection. The transfected rMSC were grown to 80% confluence and then cultured in myogenic or adipogenic differentiation medium. Cells were characterized by light microscopy, immunofluorescence and RT-PCR at different time points after myogenic or adipogenic introduction. RESULTS: Ectopic expression of activated beta-catenin located primarily in the nucleus and activated transcription in rMSC. Overexpression of stabilized beta-catenin induced 27.1 +/- 3.91% rMSC forming long multinucleated cells expressing MyoD, myogenin, desmin and myosin heavy chain (MHC) via evoking the expression of skeletal muscle-specific transcription factors. In addition, overexpression of activated beta-catenin inhibited the adipogenic differentiation in rMSC through down-regulated expressions of C/EBPalpha and PPARgamma. DISCUSSION: To our knowledge, this is the first evidence that activated beta-catenin can induce myogenic differentiation in rMSC. The ability of stabilized beta-catenin to induce myogenic differentiation in rMSC may allow for its therapeutic application. PMID- 17917886 TI - A novel high-yield volume-reduction method for the cryopreservation of UC blood units. AB - BACKGROUND: For the application of umbilical cord blood (UCB) units as hematopoietic grafts, a dose of 3.7 x 10(7) nucleated cells (NC)/kg body weight is required. NC can be lost during volume-reduction processing and during thawing. A novel modification of the double-processing protocol with the aim of minimizing NC loss is described and evaluated. METHODS: One-hundred and fifty UCB were collected. The volume was reduced by a centrifugation step following double processing in the presence of 2% HES 200/0.5. Pre- and post-processing cell counts and platelet parameters were measured with an automatic counter. The number of viable CD34+ hemopoietic stem cells was measured by flow cytometry. In 25 of the samples, colony-forming units (CFU) were also determined. The same samples were thawed 6 months after cryopreservation and re-evaluated. RESULTS: The volume was reduced to 6 +/- 1.5 mL. The recovery of NC, MNC, CD34+ hemopoietic stem cells, RBC depletion and CFU following double-processing was 93.6 +/- 3.2%, 95.8 +/- 2.2%, 98.4 +/- 1.5%, 96.8 +/- 1.1% and 107.1 +/- 6.1% (for 25 samples), respectively. The post-thaw recoveries of NC, MNC, CD34+ hemopoietic stem cells and CFU (for 25 samples) were 78.6 +/- 5.4%, 90.8 +/- 4.4%, 96.4 +/- 2.5%, 89.1 +/- 4.1%, respectively. No post-thaw cell aggregation was observed. A significant (P<0.05) post-thaw loss of platelets and signs of platelet activation was observed. DISCUSSION: The protocol uses non-expensive equipment and clinically approved materials and results in samples that can be used in patients with a mean weight of 32.7 kg. PMID- 17917887 TI - Phase I study of tumor Ag-loaded IL-12 secreting semi-mature DC for the treatment of pediatric cancer. AB - BACKGROUND: Cancer vaccines employing DC in their capacity as APC have been tolerated well and have shown some efficacy in clinical studies. IL-12, a cytokine critical for type 1 T-helper (Th1) lymphocyte and cytotoxic T-lymphocyte (CTL) differentiation, when released from a DC-based cancer vaccine, may support the generation of a cellular T-cell response. METHODS: We applied tumor cell lysate plus keyhole limpet hemocyanin (KLH)-loaded and 48-h lipopolysaccharide (LPS) plus IFN-gamma-stimulated fully mature DC, which do not release IL-12, subcutaneously to eight patients, and maximally 6-h stimulated semi-mature (sm) DC, which are potent producers of IL-12, subcutaneously (n=6) or intranodally (n=8) as a cancer vaccine to patients suffering from advanced solid pediatric malignancies. RESULTS: No serious adverse events were observed following application of IL-12-releasing smDC. Following immunization the majority of patients responded positively to KLH in a delayed-type hypersensitivity (DTH) test. In addition, three of six intranodally treated patients responded to the tumor Ag in the DTH test. DISCUSSION: We conclude that treatment with a DC-based cancer vaccine enabled to release the immune regulatory cytokine IL-12 is safe and feasible and has the potential to induce a cellular immune response in pediatric cancer patients. PMID- 17917888 TI - Adipose-derived stem cells and chondrogenesis. AB - Cartilage has only a very limited capacity to renew its original structure. Stem cells have been used to repair damaged cartilage, and recent studies have indicated that stem cells from adipose tissue are attractive cell sources that have the capacity of multipotentiality to differentiate into osteogenic, chondrogenic, myogenic, neurogenic and endothelial cells. Adipose-derived stem cells (ASC) have unique characteristics compared with stem cells from BM. At present, ASC have been studied to promote chondrogenesis. This review discusses the application of ASC to cartilage formation. PMID- 17917889 TI - Four cases of donor cell-derived AML following unrelated cord blood transplantation for adult patients: experiences of the Tokyo Cord Blood Bank. PMID- 17917890 TI - Recombinant expression of coagulation factor VIII in hepatic and non-hepatic cell lines stably transduced with third generation lentiviral vectors comprising the minimal factor VIII promoter. AB - BACKGROUND: Lentiviral vectors have the capacity to transduce stably non dividing, differentiated and undifferentiated cells of various tissues, including liver. To obtain high-level expression of transgenes, vectors often rely on viral promoters. However, recent data suggest that the supraphysiologic expression from ubiquitous viral promoters may not be beneficial and harbor the risk of oncogene activation. Therefore this study explored the lentiviral-mediated expression of human coagulation factor VIII (FVIII) driven by the physiologic FVIII gene promoter (FVIII-p), the liver-specific human alpha-1-antitrypsin gene promoter (hAAT-p), the ubiquitous but non-viral EF1alpha promoter (EF1alpha-p) and the viral CMV promoter. METHODS: Hepatic and non-hepatic cell lines were stably transduced with lentiviral vectors encoding FVIIIdelB and EGFP. To compare the different promoters, lentiviral vectors were cloned to drive FVIII expression from FVIII-p, EF1alpha-p, hAAT-p and CMV-p. RESULTS: As expected, the strong viral CMV-p and the ubiquitous EF1alpha-p resulted in the highest FVIII expression in all cell lines tested (CMV-p 1.85 IU/mL/10(6) cells for 293T, 3.15 for HepG2, 5.03 for SK-Hep, 0.91 for Hepa1-6; EF1-alpha promoter 0.30 IU/mL/10(6) cells for 293T, 0.04 for HepG2, 2.75 for SK-Hep, 0.46 for Hepa1-6). While the hAAT-p resulted in low FVIII levels (0.10 IU/mL/10(6)cells in HepG2 and 0.04 in Hepa1-6), the FVIII promoter gave reasonable expression levels in hepatic cells (0.47 IU/mL/10(6)cells in Hepa1-6 and 0.44 in SK-Hep). DISCUSSION: These results indicate the potential usefulness of the FVIII-p for hemophilia A gene therapy. PMID- 17917891 TI - UC blood-derived mesenchymal stromal cells: an overview. AB - The UC is a readily available source of blood that may be used for analysis and treatment. Some authors suggest that within the UC blood (UCB) are cells with potential for differentiation down mesenchymal lineages. Isolation and characterization of these cells has been accomplished in some centers. Differentiation of these cells down multiple lineages has been documented. Surface marker expression and gene expression profiling has been performed, and mesenchymal stromal cells (MSC) from BM and adipose tissue have been compared with those derived from UCB. The use of UCB-derived stem cells has been investigated in pre-clinical studies. As this field is rapidly advancing, this review summarizes the current state of our knowledge of MSC derived from UCB. PMID- 17917893 TI - Nucleotide sequence of a G/11 family xylanase encoding gene in Scytalidium thermophilum. AB - The nucleotide sequence of the xylanase encoding gene in Scytalidium thermophilum Af101-3 was determined. The gene encodes a family G/11 xylanase, and the coding region is interrupted by a 72 bp intron. Transcription of the gene was investigated by reverse transcription PCR (RT-PCR). Transcription of the gene was not affected by the presence of 2% glucose in the medium. Xylanase production in S. thermophilum Af101-3 was also affected by concentration of glucose in the medium (modified Czapek's supplemented with 2% corn cob powder and 0.1% glucose). Therefore, xylanase expression in this fungus may not be regulated by the carbon source in the medium. PMID- 17917894 TI - Ergonomics in schools. AB - This Special Issue on Ergonomics in Schools is dedicated to the memory of my co guest editor Cheryl Bennett who sadly and unexpectedly passed away peacefully on 2 July 2007 - only 10 days before this editorial was completed. Cheryl was inspirational in her vigorous championing of this new area of ergonomics activity, most prominently and to great effect through her foundation chairmanship of the International Ergonomics Association (IEA) Technical Committee on Ergonomics for Children in Educational Environments (ECEE) (see www.iea.cc). PMID- 17917892 TI - A large-scale method for the selective depletion of alphabeta T lymphocytes from PBSC for allogeneic transplantation. AB - BACKGROUND: We sought to develop a method for the clinical large-scale depletion of alphabeta T lymphocytes from mobilized peripheral stem cells, which would allow the allogeneic transplantation of a graft enriched for stem cells, natural killer (NK) cells and gammadelta T lymphocytes. METHODS: Therefore, we obtained mononuclear cells from either mobilized or non-mobilized healthy adult volunteer donors and incubated the cells with a biotinylated anti-alphabeta T-cell Ab and subsequently with an anti-biotin Ab conjugated with magnetic microbeads. The depletion was then performed using a CliniMACS device. RESULTS: The median T-cell depletion was 3.9 log (range 3.5-4.1 log). The recovery of the gammadelta and NK cells was 92% and 80%, respectively. The recovery of CD34+ stem cells from the mobilized donors was 66%. DISCUSSION: This method had no negative influence on the in vitro colony formation of stem cells, and transplantation of alphabeta depleted cells into NOD-SCID IL-2 common gamma chain knockout (NOD-scid IL2r (null)) mice resulted in a rapid engraftment of human myeloid and lymphoid cells. This method will allow large-scale depletion of alphabeta T cells from mobilized peripheral blood in clinical trials. PMID- 17917895 TI - The ergonomics of learning: educational design and learning performance. AB - The application of ergonomics/human factors (E/HF) principles and practices, and the implementation of ergonomics programmes, have achieved proven success in improving performance, productivity, competitiveness, and safety and health in most occupational sectors. However, the benefits that the application of E/HF science might bring to promoting student learning have yet to be widely recognized. This paper deals with the fundamental purpose of education - student learning - and with the question of how the ergonomic design of the learning environment influences learning performance. The underlying premise, embodied in the quote below, is that student learning performance to a substantial degree is context specific - influenced and specialized in relation to specific design factors in the learning environment. The basic scientific question confronting learning ergonomics is which design characteristics in the learning environment have the greatest influence on variability in learning performance. Practically, the basic challenge is to apply this scientific understanding to ergonomic interventions directed at design improvements of learning environments to benefit learning. This paper expands upon these themes by addressing the origins and scope of learning ergonomics, differing perspectives on the nature of learning, evidence for context specificity in learning and conclusions and research implications regarding an ergonomics perspective on learning. PMID- 17917896 TI - Ergonomics, education and children: a personal view. AB - Educational ergonomics - the teaching of ergonomics and the design of environments where ergonomics teaching and learning might occur - has received little attention from ergonomists. This paper first describes the roots of the author's interest and research in educational ergonomics; second it provides a personal view of the opportunities and challenges posed by the two streams of educational ergonomics; and lastly it considers the implications of teaching ergonomics to children in terms of their personal development, the design of schools and the impact such initiatives might have on wider societal problems. PMID- 17917897 TI - Are the desks and chairs at school appropriate? AB - The aim of the current study was to find out how the measures of chairs and desks match with the anthropometrics of schoolchildren and how schoolchildren sit during a lesson in their classroom. This paper reports the baseline measurements of an intervention study. Participants of this study were 6th and 8th grade (12 and 14 year old) schoolchildren from two comprehensive schools in Finland (N = 101, 57 girls and 44 boys). The main outcome measures were the differences between desk height and elbow-floor height, and chair height and popliteal height. Forty-three participants were randomized for sitting posture analysis by video recordings. The study showed that desks were on average 13 cm above elbow floor height and chairs 2 cm below popliteal height. For 56% of time participants sat with their backs flexed >20 degrees and/or rotated >45 degrees . For 70% of time they sat with their necks flexed >20 degrees or rotated >45 degrees. The results indicate that there is a mismatch between school furniture and the anthropometrics of schoolchildren. Schoolchildren sit in disadvantaged postures for a substantial part of school lessons. PMID- 17917898 TI - Classroom postures of 8-12 year old children. AB - This study examined classroom postures of 8-12 year old school children in Flanders and related the outcomes to self-reported back or neck pain. Postural behaviours using the portable ergonomic observation (PEO) method and self reported one-week back and neck pain were studied in 105 children from 41 different class groups. Pupils sat statically for 85% of the time, 28% of which the trunk was bent or flexed forward. For 9% of the time, children sat dynamically and for 36% they used a back rest. Children who spent more time sitting with a flexed trunk reported significantly more thoraco-lumbar pain compared to pain-free children and to children with cervical pain (p < 0.05). Children reporting pain stood for a longer period of time than pain-free children (p < 0.05). It is concluded that prolonged static kyphotic sitting without use of a backrest is common in elementary school children in Flanders. PMID- 17917899 TI - An investigation of children's posture and discomfort during computer use. AB - This study investigated schoolchildren's posture and discomfort while working at computers. Sixty-eight children (mean age 9.5 years) were observed at school during normal computer sessions lasting 15-25 min. Rapid upper limb assessment (RULA) was used to evaluate posture, and a body discomfort chart (BDC) and a modified visual analogue scale (VAS) were used to record site and intensity of discomfort. Computer tasks were noted and in accordance with RULA, postures were classified as Action Level (AL) 1 (acceptable) to 4 (needs immediate change). Most children adopted postures at an unacceptable level while working at computers. None of the postures were in AL 1; 60% were in AL 2; 38% were in AL 3; and 2% were in AL 4. Posture became worse over time. Poor posture was associated with discomfort, but it is not clear if it was related to the sitting posture or to the computer use. Children who reported discomfort had a higher mean RULA grand score (5.0) than those who did not report discomfort (4.4). The type of computer task influenced the children's posture. RULA proved generally to be a suitable method for evaluating children's posture. PMID- 17917900 TI - Seated buttock contours: a pilot study of Australian senior high-school students. AB - Both posture and comfort of a chair are influenced by the contour and characteristics of the seat. Knowledge of seat contours of a student population could thus be useful in the design of school chairs. This study investigated seated buttock contours of senior high-school students in order to determine: (a) their general characteristics, (b) the effect of gender and sitting posture and (c) the relationship between the contours and selected anthropometric variables (stature and mass). A contour measurement device was developed and used to measure buttock contours in five sitting postures (typing, sitting up, sitting back, slumping and writing). Buttock contours were quantified by constructing anterior-posterior (AP) and lateral profiles from which six discrete profile dimension measurements were made. AP and lateral profiles were found to have a consistent shape across all participants. Five out of six profile dimensions were significantly different between genders, with just one significantly different between sitting postures (typing and sitting back). Correlations between anthropometric measures and profile dimensions were relatively low (r < 0.34) with no clear patterns evident. Overall results of this study suggest that buttock contours are influenced by gender to a greater extent than sitting posture. PMID- 17917901 TI - Preferred seat orientation of senior high-school students. AB - The height of the front of the seat is the primary determinant of appropriate seat size in the school setting. In the present study, this dimension was fixed at 445 mm and, using a brief adjustment period, students adjusted the angle of the seat to their preferred rear seat height (PRSH) under three fixed and one adjustable desk height conditions and for one desk height, under two time conditions. PRSH was significantly greater at an 800 mm desk height (454+/-14 mm) compared to 735 mm (447+/-15 mm) and 720 mm (444+/-16 mm). When desk height as well as rear seat height were adjustable, PRSH was 446+/-15 mm and preferred desk height 751+/-25 mm. Taller students or those with larger popliteal heights selected lower PRSHs at all desk heights, with PRSH more strongly related to popliteal height (r = -0.54 to -0.59) than stature (r = -0.44 to -0.50). No differences were found in PRSH between short (<5 min) and long (30 min) adjustment periods for the 735 mm desk height. The nearly horizontal seat positions found in this study were between those recommended by other authors. PMID- 17917902 TI - Lack of conformity between Indian classroom furniture and student dimensions: proposed future seat/table dimensions. AB - Children spend one-quarter of a day in school. Of this, 60-80% of time is spent in the classroom. Classroom features, such as workspace and personal space play an important role in children's growth and performance as this age marks the period of anatomical, physiological and psychological developments. Since the classroom is an influential part of a student's life the present study focused on classroom furniture in relation to students' workspace and personal space requirements and standards and was conducted in five schools at Mumbai, India. Dimensions of 104 items of furniture (chairs and desks) were measured as were 42 anthropometric dimensions of 225 students from grade six to grade nine (age: 10 14 years). Questionnaire responses of 292 students regarding the perceived adequacy of their classroom furniture were collected. Results indicated that the seat and desk heights (450 mm, 757 mm respectively) were higher than the comparable students' anthropometric dimensions and that of the recommendations of Bureau of Indian Standards (BIS) (340 + 3 mm, 380 + 3 mm seat-heights, 580 + 3 mm 640 + 3 mm desk-heights) as well as Time-Saver Standards (TSS) (381.0 mm seat height and 660.4 mm desk-height). The depth of the seats and the desks (299 mm, 319 mm, respectively) were less than comparable students' anthropometric dimensions and the recommendations of BIS (IS 4837: 1990). Students reported discomfort in shoulder, wrist, knee and ankle regions. Based on the students' anthropometric data, proposed future designs with fixed table-heights and adjustable seat-heights along with footrests were identified. PMID- 17917903 TI - Definition of sizes for the design of school furniture for Bogota schools based on anthropometric criteria. AB - The current paper deals with the definition of sizes for the design of school furniture for schools in Bogota, Colombia, based on an analysis of available anthropometric data on Latin American children. State-of-the-art anthropometric, national and international standards were considered, in order to define the anthropometric variables that were to be used for defining the furniture. Matrices relating age and specific anthropometric dimensions were constructed, as a visualization method for establishing the dimensional differences between children of the same age and the ranges that should be covered by the items of furniture. Dimensional data were grouped by establishing the minimum sizes and general dimensions of furniture needed to cover the 5-95th percentile of school children between the ages of 5 and 18 years. The distribution of the furniture in the different school grades was also indicated. Apart from the need for an adequate match between child anthropometry and school furniture dimensions, this study shows the importance of a proper distribution of furniture sizes in the different school grades, as a complementary and decisive aspect to be considered in order to meet the heterogenic, anthropometrical requirements of children of the same age and school grade. PMID- 17917904 TI - Sitting and standing postures are corrected by adjustable furniture with lowered muscle tension in high-school students. AB - This study compared the effect of 24 months of adjustable school desks and chairs usage (the intervention) and traditional non-adjustable usage (the control condition) on sitting and standing postures, muscle strength, classroom muscle tension, pain and learning in 15 (8 female and 7 male) high-school students and 15 anthropometrically and gender matched control students from neighbouring schools. It was assessed whether any responses took place after growth cessation. In comparison with controls, the intervention group of students' sitting postures standing kyphosis, scoliosis and lordosis became significantly better, both before and after growth cessation. Trunk muscle strength increased in the intervention students whose muscle tension during classes fell significantly in the trapezius and lumbar muscles, whereas in control students' lumbar tension increased. Headache and low-back pain correlated with neck-shoulder pain and trapezius muscle tension. Intervention students reported that they experienced benefits from the adjustable tables and chairs. They also received significantly better overall marks than the controls at the end of high school. It is concluded that the adjustable school desks and chairs promoted better sitting and standing postures, increased muscle strength, alleviated pain and appeared to be associated with better overall academic marks. PMID- 17917905 TI - School children's use of computers and teachers' education in computer ergonomics. AB - A national survey to investigate the education of teachers in computer-related ergonomics was carried out by postal questionnaire. The use of computers by primary school children (age 4-12 years) was also investigated. Data were collected from a random sample of 25% (n = 830) of primary schools in the Republic of Ireland. Questionnaires (n = 1863) were returned from 416 schools giving a response rate of 50.1%. Almost all schools (99.7%) had computers for children's use. The computers were most often (69.8%) used in the classroom. The majority (56.3%) of children worked in pairs. Most teachers (89.6%) had received computer training, but few (17.6%) had received ergonomics information during the training. Respondents were not satisfied with their current knowledge of ergonomics. Over 90% stated that they would like to receive further information by printed format or during a training course, rather than by computer (web or CD ROM). PMID- 17917906 TI - Measurement of the temporal patterns of school bag carriage using activity monitoring and structured interview. AB - The primary objective of the current study was to quantify the temporal patterns of school bag carriage over an actual school day using activity monitoring and structured interviews, in order to better understand the physical demands of school bag carriage. The temporal patterns of 40 students' school bag carriage over a 24-h period were defined by total school bag carrying time, mean event school bag carrying time, the number of school bag carrying events, total carrying time travelling to and from school and the number of students who walked or used transport to travel to and from school. There were significant correlations between activity monitor [mean(SD) 119(48) min] and structured interview [100(39) min] determined total school bag carrying time (r = 0.59), activity monitor [8(4) min] and structured interview [9(4) min] determined mean event school bag carriage time (r = 0.65), and activity monitor [15(4) events] and structured interview [11(2) events] determined number of school bag carrying events (r = 0.52). However, the number of school bag carrying events, and for students who used transport, the total amount of time spent travelling to school was significantly different using the two measures. The durations of school bag carriage and the relationship between activity monitor and structured interview were similar to those reported in previous studies. PMID- 17917907 TI - Recurrent non-specific low-back pain in adolescents: the role of exercise. AB - This study evaluated the efficacy of an exercise programme as an intervention for recurrent non-specific low-back pain (NLSBP) in adolescents. A randomized controlled trial was conducted with an experimental group (n = 27, age 14.6 years) who participated in an 8-week exercise programme and a matched control group (n = 27, age 14.6 years) who continued normal daily activities. All participants suffered from recurrent NSLBP. Pre and post intervention measures of NSLBP status (pain severity and consequences) and daily inactivity (time spent sitting, PC time, TV time) were reported in one week diaries. Two-way mixed ANOVA (independent variables: pre/post and experimental/control) was conducted for each dependent variable, significance was set at P < 0.05. Significant interaction effects were identified for the severity of pain, number of occasions missing sport due to NSLBP and amount of sport participated in. In each case the experimental group benefited from the exercise programme. In contrast, no significant interaction effects were observed for physical inactivity, both groups spent a similar amount of time sitting, watching TV and using a PC pre- and post- intervention. It was concluded that an exercise programme acted as an effective short-term treatment strategy for NSLBP in adolescents. Further evaluation is required to assess the long-term effectiveness. PMID- 17917908 TI - Metabolic rate and clothing insulation data of children and adolescents during various school activities. AB - Data on metabolic rates (n = 81) and clothing insulation (n = 96) of school children and adolescents (A, primary school: age 9-10; B, primary school: age 10 11 year; C, junior vocational (technical) education: age 13-16 (lower level); D, same as C but at advanced level; and E, senior vocational (technical) education, advanced level: age 16-18) were collected (Diaferometer, Oxylog, Heart Rate derivation) during theory-, practical- and physical education- lessons. Clothing insulation was calculated from clothing weight, covered body surface area, and the number of clothing layers worn. Clothing insulation was found to be similar to that expected for adults in the same (winter) season, with minimal variation with age or school type (0.9 to 1.0 clo; 1.38 clo where coverall was worn), but more variation within groups (coefficient of variation 6-12%). Metabolic rate values (W.m(-2)) were lower than expected from adult data for similar activities, but are supported by other child data. The results of this study can be used to establish design criteria for school climate control systems or as general data on energy expenditure for children and adolescents. The results emphasize the need for specific child data and show the limited value of size-corrected adult data for use in children. PMID- 17917909 TI - Road-safety education: spatial decentering and subjective or objective picture processing. AB - The current study examined children's ability to analyse pictures of a risky situation, both in relation to the characteristics of the pictures and in relation to the centering/decentering process of cognitive development. Sixty children aged 6, 9 or 11 years were given an objective or subjective version of a story about a risky situation involving road crossing and were asked to reconstruct it by putting six pictures in chronological order. The type of picture series, objective or subjective, had a different effect on the children's understanding and performance, according to the age. The older children were better at ordering the pictures, but on the subjective version only. The picture version effect on planning time decreased with age; only the younger children took more time to start touching the pictures. On one hand, it is concluded that for the youngest children, objective representations are essential to analysing pictures showing a risk, whereas the oldest children will profit more from a subjective view. On the other hand, subjective representations, which give a more realistic view, provide an excellent tool for testing children's abilities. Subjective representations can be used to detect potentially risky behaviour in virtual situations (static pictures, or multimedia tools), since it permits one to predict at-risk behaviour in the street and to assess the effectiveness of remedial measures. PMID- 17917911 TI - Mycotoxin occurrence and Aspergillus flavus soil propagules in a corn and cotton glyphosate-resistant cropping systems. AB - The effects of cotton-corn rotation and glyphosate use on levels of soil-borne Aspergillus flavus, aflatoxin and fumonisin contamination in corn and cotton seed were determined during 2002-2005 in Stoneville, Mississippi (USA). There were four rotation systems (continuous cotton, continuous corn, cotton-corn and corn cotton) for both glyphosate-resistant (GR) and non-GR cultivars-herbicide system arranged in a randomized complete block design with four replications. Aspergillus flavus populations in surface (5-cm depth) soil, sampled before planting (March/April), mid-season June) and after harvest (September), ranged from 1.47 to 2.99 log (10) cfu g(-1) soil in the four rotation systems. Propagules of A. flavus were higher in the continuous corn system compared to the continuous cotton system on three sample dates, and cotton rotated with corn decreased A. flavus propagules in three of nine sample dates. Propagules of A. flavus were significantly greater in plots with GR cultivars compared to non-GR cultivars in three samples. In cotton seed, aflatoxin and fumonisin levels were similar (< or = 4 microg kg(-1) and non-detectable, respectively) regardless of rotation and glyphosate. In corn grain, aflatoxin was above the regulatory level (> or = 20 microg kg(-1)) only in GR cultivar in 2004 and 2005. Fumonisin was higher in non-GR cultivar (4 mg kg(-1)) regardless of rotation in 2004; however, in 2002, 2003 and 2005, aflatoxin and fumonisin levels were similar regardless of rotation and glyphosate. These results indicate the potential for increased aflatoxin and fumonisin levels (1 of 4 years) in corn; however, climatic conditions encountered during this study did not allow for mycotoxin production. In laboratory incubation studies, fairly high concentrations of glyphosate were required to inhibit A. flavus growth; however no short-term effect of soil treatment with glyphosate on A. flavus populations were observed. These data suggest that altered populations of A. flavus or higher aflatoxin concentrations in corn grain were due to indirect effects of the GR cropping system. PMID- 17917912 TI - Preparation of anionic poly(epsilon-caprolactone)-poly(ethylene glycol) poly(epsilon-caprolactone) copolymeric nanoparticles as basic protein antigen carrier. PMID- 17917913 TI - Changes of glycoconjugate expression in nasal respiratory mucosa of rats exposed to welding fumes. AB - To investigate the effects of welding fumes on the glycoconjugates in nasal respiratory mucosa, male Sprague-Dawley rats were exposed to manual metal arc stainless steel (MMA-SS) welding fumes at a concentration of 56-76 mg/m(3) total suspended particulate for 2 h/day in an inhalation chamber for 90 days. During the exposure period, the experimental animals were sacrificed after 2 h and 15, 30, 60, and 90 days of exposure; then sections were examined using lectin histochemistry. Some remarkable changes, such as destroyed cilia, desquamation and mucification of epithelial cells, and destruction of nasal septal glands, were seen in the welding fume-exposed groups. Specific changes in the lectin binding patterns were also observed in the welding fume-exposed rats. The Ricinus communis agglutinin-I (RCA-I) staining of the cilia and columnar cells increased slightly when compared with the unexposed rats. The RCA-I and Ulex europaeus agglutinin-I (UEA-I) staining of the goblet cells also increased as the exposure continued. The mucigenous epithelial cells reacted with Bandeiraea simplicifolia lectin-I (BSL-I), RCA-I, and succinylated wheat germ agglutinin A (sWGA) after 15 days of exposure, which was not visible in the control group. The dorsal septal glands exhibited an affinity with peanut agglutinin (PNA), BSL-I, and RCA-I, which was also not visible in the control group. The affinity for Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), PNA, sWGA, BSL-I, and UEA-I in the ventral septal glands of the welding fume-exposed groups tended to increase, whereas the concanavalin A (Con A) reactivity in the dorsal septal glands decreased slightly. In conclusion, it was assumed that the changes in the glycoconjugate residues in the nasal respiratory mucosa of the welding fume exposed rats represented important components of defense mechanisms against the toxicants in the welding fumes. PMID- 17917915 TI - Radiological progression and mortality among silica flour packers: a longitudinal study. AB - There are few longitudinal studies that investigated the radiological progression of silicosis among different occupational groups. Inhalation exposure to very high levels of crystalline silica can lead to acute silicosis. This longitudinal study was designed to determine association between mortality and radiological progression among individuals with rapidly progressive silicosis. Twenty-three subjects with silicosis had previously worked in the same stone-grinding factories, where they had been exposed to high airborne concentrations of silica powder dust. The diagnosis of rapidly progressive form of silicosis was made according to the National Institute for Occupational Safety and Health definition. The subjects were entered into the study if chest radiography films with acceptable quality showed small opacity shadowing of at least 1/0 profusion grade. Chest radiographs of the start and end of follow-up were independently reviewed at different times by two readers. For paired comparison, the side-by side method of presenting radiographs was used. Data analysis was carried out by using the t-test, Fisher's exact test, and logistic regression. It was found that 65.2% of workers showed evidences of radiological progression during a period of follow up (mean 30.3 mo). The age of death for the 10 workers who died of silicosis ranged between 21 and 35 yr, with the mean and median age of 27.30 (SD 4.77) and 28.50 yr, respectively. There was a significant association between mortality and both size and profusion progression of small opacities. In conclusion, we quantified the risk of death from rapidly progressive forms of silicosis in a group of workers heavily exposed to silica. We conclude that young adult age, rapid progression in the size, and profusion of small opacities are risk factors for the development of advanced silicosis. Death occurs more quickly in silicotic patients with these criteria. PMID- 17917914 TI - Immunotoxicological study of one of the most common over-the-counter pyrethroid insecticide products in Egypt. AB - A commonly available aerosolized pyrethroid insecticide containing deltamethrin and imiprothrin is widely used for hygienic control in Egypt. The immunotoxic effects after inhalation exposures to the preparation of each for 2, 10, and 30 days were investigated in rats. For each exposure, the insecticide (containing 0.2% imiprothrin and 2.5% deltamethrin) was sprayed in all directions in a room (using a special attachment located in the ceiling in the center of the room) for 30 s each minute for 15 min; the room was then kept closed for 15 min. After each spray interval, the rats were introduced for 30 min and then removed to a clean room. The exposure process was repeated a total of three times on each day of the respective regimens. The interval between the 15-min spray/15-min pause/30-min rat exposure cycles was 120 min. Twenty-four hours after the final exposure in each particular regimen, the cohort rats in the regimen (air and exposed) were weighed, sacrificed, and their tissues were removed for analyses. Immunological tests performed included assessments of potential changes in immunopathology (determined from body and splenic weights), humoral-mediated immunity (based on plaque-forming activity of spleen cells), cell-mediated immunity (determined from splenic lymphocyte responsiveness to stimulation with phytohemagglutinin and immune cell (sub)type profile analyses), and nonspecific immunity (based on phagocytic activity of peritoneal macrophages). The results indicated that of all the endpoints examined, among the rats exposed over a 2-day period to the imiprothrin- and deltamethrin-containing insecticide aerosol, the only significant change noted (relative to values from time-matched controls) was in the levels of splenic CD4+CD8- and CD4+ CD8+ cells. In contrast, exposures on each day of a 10-day period led to significant decreases in several endpoints; exceptions to this were values for body and spleen weight (unaffected), splenic OX12-OX19+ levels (significant increase), and CD4+CD8- levels (unaffected, relative to control). Rats exposed for 30 days displayed significant decreases in each test applied, except for increases in both splenic OX12-OX19+ and CD4+CD8- cell levels relative to corresponding control rat values. The present study findings indicate that repeated noncontinuous inhalation of a commonly utilized insecticide that contains imiprothrin and deltamethrin can cause a variety of immunotoxic effects in sites distal to the lungs. PMID- 17917917 TI - Traffic, air pollution, and health. AB - This paper discusses emerging information on exposure to air pollution from traffic and health and raises two key questions with regard to the impact of traffic on public health: 1) is there sufficient evidence to infer that traffic is causing adverse health effects, and; 2) what is the magnitude of the adverse impact of traffic on health? Topics addressed include characterization of exposure to traffic-related pollutants; the findings of epidemiological studies to date; and investigative approaches and the related challenges - including bias, model-based conclusions, and sample size issues--in characterizing the health effects of air pollution from traffic. Also considered are the known health effects of two of the major pollutants produced by vehicle exhaust- particles and ozone. The evidence points to traffic as a threat to public health that will be managed with great difficulty; however, more research is needed to refine our understanding of the health consequences of traffic exposures and as a basis for formulating mitigation policies. This paper discusses emerging information on exposure to air pollution from traffic and health and raises two key questions with regard to the impact of traffic on public health: 1) is there sufficient evidence to infer that traffic is causing adverse health effects, and; 2) what is the magnitude of the adverse impact of traffic on health? Topics addressed include characterization of exposure to traffic-related pollutants; the findings of epidemiological studies to date; and investigative approaches and the related challenges - including bias, model-based conclusions, and sample size issues - in characterizing the health effects of air pollution from traffic. Also considered are the known health effects of two of the major pollutants produced by vehicle exhaust - particles and ozone. The evidence points to traffic as a threat to public health that will be managed with great difficulty; however, more research is needed to refine our understanding of the health consequences of traffic exposures and as a basis for formulating mitigation policies. PMID- 17917918 TI - Biodiesel: production and economic issues. AB - The convergence of a number of social, economic, and political developments, at both the national and global levels, has created much interest in developing biodiesel as an alternative to petroleum diesel. In response to higher energy prices, along with government incentives, the fledgling biodiesel industry is beginning to grow at a rapid pace. However, there are clearly economic limits to biodiesel's growth potential and questions are being raised about the future role biodiesel can play in reducing our energy dependence Since vegetable oils and animal fats will continue to be used primarily for food products, biodiesel production can only supply a small portion of our fuel needs. To further increase biodiesel production, we will need to develop new sources of feedstock and develop new technologies to convert a broader spectrum of feedstocks. PMID- 17917919 TI - The impact of biodiesel on pollutant emissions and public health. AB - An overview of recent studies of the impact of biodiesel and biodiesel blends on air pollutant emissions and health effects is provided. Biodiesel blends of 20% produce reductions of 15% or higher (depending upon engine model and test cycle) in emissions of particulate matter, carbon monoxide, total hydrocarbons, and a group of toxic compounds including vapor-phase hydrocarbons from C1 to C12, aldehydes and ketones up to C8, and selected semivolatile and particle-phase PAH and NPAH. Based on the studies reviewed and recently acquired data, individual engines may show oxides of nitrogen increasing or decreasing, but on average there appears to be no net effect for blends of 20% biodiesel--the most common biodiesel blend. Exhaust from a diesel engine operating on 100% biodiesel was also shown to have only modest adverse effects in an animal exposure study. Studies of the impact of biodiesel on particle size have not produced consistent results and additional research in this area is needed. Biodiesel is also shown to significantly reduce life-cycle greenhouse gas emissions in comparison to petroleum diesel. PMID- 17917920 TI - The use of biodiesel in a school transportation system: the case of Medford Township, New Jersey. AB - A combination of high fuel prices, bus maintenance costs, and the health and safety of school children, along with a consideration of federal and state regulations, prompted Medford Township school district in southern New Jersey to explore the use of alternative fuels, specifically biodiesel. The school district owns and operates 62 school buses that transport 3500 children daily. The evolution of this switch from petroleum-based fuel to biodiesel is described. The district is the nation's longest continuous user of biodiesel in a school transportation system. PMID- 17917921 TI - High interleukin-10 production is associated with anti-acetylcholine receptor antibody production and treatment response in juvenile myasthenia gravis. AB - To investigate the significance of humoral immunological factors in juvenile myasthenia gravis (JMG) patients, 11 children with JMG (age of onset between 2 and 13 years) and 8 healthy controls were enrolled. Plasma IL-2, IL-10, IFN gamma, TNF-alpha, complement factor C3, C4, and C1q levels were measured by ELISA. IL-2, IL-10, TNF-alpha, C3, and C4 levels were significantly elevated in the patient group. Moreover, plasma IL-10 levels were associated with the presence of anti-AChR antibody and response to treatment suggesting a critical role for IL-10 in JMG pathogenesis. PMID- 17917922 TI - Influence of acute hyperglycaemia on the amplitude of nociceptive spinal evoked potentials in healthy rats. AB - To evaluate the effect of blood glucose level on the amplitude of nociceptive spinal evoked potentials in healthy rats, an acute hyperglycaemia state was induced in an experimental group of 12 rats, through the infusion of glucosade solution. A Ringer-lactate solution was administered equivolumetrically to the control group (5 rats) under the same experimental conditions. Nociceptive spinal evoked potentials were recorded every 2 min, before and during the induction of hyperglycaemia, from the left lumbar cord dorsum activated orthodromically by ipsilateral electrical stimulation of the hind paw (20 Volts, 0.5 ms, 0.2 Hertz). Acute hyperglycaemia induced an increase of amplitude in both N (+8.92%, p = .000006) and P (+10.46%, p = .000037) waves when comparing control and experimental groups or basal versus infusion values, in response to nociceptive stimuli. The present results show that acute hyperglycaemia could contribute to central nociceptive sensitization; it would be attributed to an increased synchronization of spinal dorsal horn neuronal discharges. PMID- 17917923 TI - Total brain tissue sialic acid levels due to glutathione effect in experimental epilepsy. AB - Epilepsy can be described as a group of neurological disorders, characterized by recurrent episodes of convulsive seizures, loss of consciousness, sensory disturbances, abnormal behavior, or all of these. Altered glutathione metabolism in association with increased oxidative stress has been implicated in the pathogenesis of many diseases such as seizures. It is therefore reasonable to propose that sialic acid levels can be affected by this pathological state or, alternately, by seizures. The present study showed that the sialic acid levels were significantly different between the experimental groups as well as in the subgroup analysis. The results suggest that glutathione may have a neuroprotective effect by decreasing sialic acid levels in mice brain. PMID- 17917924 TI - Sexual dependency of rat sciatic nerve fiber conduction velocity distributions. AB - Gender differences, either with the structural or through with hormones, dictate how the corresponding organ or organ system responses to physiological signals. Current study aims to investigate gender dependent differences in conduction related parameters of rat sciatic nerve. Compound action potentials (CAP) were recorded via suction electrode whereas the conduction velocity distributions (CVD) were performed using the method known as collision technique in the literature. Studied CAP parameters, namely conduction velocities (CV), area of the CAPs and time required to reach the maximum depolarization (TP) have been found significantly different for female and male rats. Detailed analyses have shown that sex dependent differences were more remarkable in the right leg responses of female and male rats. Additionally, CVDs indicate that the number of fibers having CVs between 5-30 m/s is much more in male right sciatic nerve trunk when compared to age matched female rats. The present study, for the first time clearly shows that shift in the contribution of nerve fibers to lower CVs is the main causal of the sex dependent differences seen in rat sciatic nerve fibers. PMID- 17917925 TI - Heat shock protein72 protects hippocampal neurons from apoptosis induced by chronic psychological stress. AB - When exposed to nonlethal heat stress (i.e., heat shock preconditioning), HSP72 expression increased in the mammalian brain. HSP72 enhance the viability of neurons and decrease TUNEL-positive neurons under several kinds of stress (e.g., ischemic). Chronic psychological stress is a kind of stress that could cause hippocampal neuron apoptosis. But whether overexpression of HSP72 can decrease TUNEL-positive hippocampal neurons caused by chronic psychological stress is unclear. To investigate the possible protective role of HSP72 in decreasing chronic psychological stress-induced hippocampal neuron apoptosis, this study analyzed HSP72 expression, apoptotic neurons in the hippocampus of mice. Adult mice were divided into four groups unstressed group; chronic psychological stress group; heat shock stress group; heat shock preconditioning plus psychological stress group; receiving no experimental stress, chronic psychological stress, heat shock stress, heat shock preconditioning plus psychological stress separately. Mice were killed after one month, two months, or three months of stress. A three-way ANOVA (psychological stress x heat shock stress x time) revealed a significant effect of heat shock stress in increasing HSP72 expression, decreasing neuronal apoptosis in hippocampus CA3 region caused by chronic psychological stress, and showed that HSP72 protected hippocampus CA3 neurons from chronic psychological stress. PMID- 17917926 TI - Executive functioning skills in male students with social-emotional disorders. AB - This study examined the efficacy of discrete versus global measures of executive functioning in children with different levels of emotional conditions. Fifty seven male students were administered the Wisconsin Card Sort Test, the Category Test, and the Behavior Rating Inventory of Executive Functioning (BRIEF). Only the BRIEF variables of Emotional Control and Shifting Attention were statistically significant variables for students with severe emotional conditions. The relationships between attentional control, emotional regulation, and higher-level problem-solving skills were discussed in the context of discrete versus global measures of executive functioning in children with emotional conditions. PMID- 17917927 TI - Cognitive effects of short-term use of raloxifene: a randomized clinical trial. AB - Two questions regarding findings from the Women's Health Initiative are (1) What is the effect of various hormonal regimens including selective estrogen receptor modulators? and (2) Is the negative effect on cognitive functioning related to the older age (65+ years) of the women? This study addresses these two questions in a short-term randomized trial of the effects of raloxifene versus alendronate on cognition. The study found only one significant interaction where the raloxifene and alendronate group changed differently across the two testing occasions. Hence, raloxifene does not have any impact, positive or negative, on short-term cognitive functioning when compared to alendronate. PMID- 17917928 TI - Expectancy of impaired neuropsychological test scores in a non-clinical sample. AB - Two methods were addressed to evaluate the incidence of false positive impairment on tests in neuropsychological evaluation. One method used a mathematical model based on the binomial theory (Ingraham & Aiken, 1996) to offer predictions based on probability. The model sets boundaries to determine when score variations would occur by chance and is therefore useful for identifying when cases exceed established boundaries. In addition, the present study examined test performance in a non-clinical sample of 94 college students via the Halstead Reitan Neuropsychological Test Battery. Results found that 15% of the sample had impairments suggested by the Halstead Impairment Index, using criteria established by Reitan and Wolfson (1993). In addition, one-half of the sample obtained impaired scores on one or two tests. These results are consistent with what the binomial model predicted. The model would therefore serve as a useful resource for clinicians when considering the probability of impaired test performance. PMID- 17917929 TI - Myasthenia gravis accompanied by thymomas not related to foamy virus genome in Belarusian's patients. AB - The spectrum and features of neurological disorders have been changed due to the Chernobyl catastrophe in the Republic of Belarus. More recently neurologists in Belarus have noted a significant increase in the frequency of myasthenia gravis (MG) with concomitant rise in the thymomas. There is some evidence suggesting that retroviruses play a key role in the development and pathogenesis of autoimmune diseases. This study analyzed thymomas from 45 MG patients from the Republic of Belarus by using PCR and primers for two regions of FV--gag and bel-2 genes. The results showed that none of the varied thymuses from the 45 MG patients contained FV genome. No relationship can be confirmed between FV and this disease and the results suggest that no pathological link between FV and MG exists. PMID- 17917930 TI - Duplication analysis in Turkish Charcot-Marie-Tooth type 1A patients using short tandem repeat markers. AB - Charcot-Marie-Tooth (CMT) is a common inherited peripheral neuropathy with a prevalence of 1 in 2,500. CMT has two distinct forms (CMT1 and CMT2) that can be identified electrophysiologically. A 1.5 Mb tandem microduplication including peripheral myelin protein 22 gene (PMP22) on chromosome 17p11.2-12 causes CMT1A. The increased gene dosage effect of PMP22 is thought to be responsible for the pathogenesis of CMT1A. In this study, 39 Turkish CMT1A patients and 60 unrelated control samples had been examined for the duplication using polymorphic short tandem repeat (STR) markers. Seven STR marker sites (AC005838-4A-, AC0013248-9A-, AC0013248-9B-, D17S2218, D17S2220, D17S2227, and D17S2229) on the duplicated region were amplified via polymerase chain reaction, electrophoresed through 8% polyacrilamide gel and evaluated for the duplication. The rate of duplication was 92.3' (36/39) in the patients whereas it was zero in the control samples. Allele distributions, number of alleles and heterozygosity values of more informative markers (D17S2218, D17S2220, D17S2227, and D17S2229) were assessed. It is found that approximately 85% of duplications in Turkish CMT1A patients were depicted by using D17S2220 and D17S2229 markers together. PMID- 17917931 TI - Early preweaning methamphetamine and postweaning rearing conditions interfere with the development of peripheral stress parameters and neural growth factors in gerbils. AB - Adrenal steroid hormones and neuronal growth factors are two interacting systemic factors that mediate the environment's influence on the brain's structure and function. In order to further elucidate their role and relationship in the effects of early stressful experience and isolated rearing (IR), this study measured blood corticosterone titres and relative adrenal weights and assessed nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) concentrations in brain regions of both hemispheres of young adult Mongolian gerbils injected on postnatal day 14 with a single high dose of methamphetamine (MA) or saline and raised after weaning either in an enriched or an impoverished environment. Irrespective of MA challenge, IR decreased corticosterone titres to about half, but increased relative adrenal weights. BDNF concentrations were decreased by IR in saline-injected animals in the left prefrontal and parietal cortices and right entorhinal and hippocampal cortices, and in the subcortical regions of both hemispheres. NGF concentrations were unaltered by IR in saline injected animals, but increased in MA challenged animals in the entorhinal/hippocampal cortices and subcortical areas of both hemispheres. MA application induced shifts of the lateral asymmetry in NGF contents in prefrontal and entorhinal cortices. The results suggest that an early pharmacological traumatization can set a switch for further brain development, and that growth factor concentrations might possibly be influenced by peripheral stress hormones. PMID- 17917933 TI - Gender inequalities in maternal perception of healthy child body shape and their association with child body mass and blood pressure. AB - OBJECTIVE: To investigate whether maternal perception of child body shape is different for boys and girls and whether this subjective parameter is associated with objective indices of cardiovascular health. DESIGN: Cross-sectional survey (the ARCA Project). SETTING: Southern Italy. SUBJECTS: Body mass index, waist circumference and blood pressure were measured according to standardised procedures in 3551 children (Boys = 1778; Girls = 1773) aged 6-11 years. Maternal perception of child body shape was investigated by inviting mothers to choose, within two sets of seven, 9-year-old child silhouettes (one set for boys and one set for girls), the body shape they perceived as that of a healthy child. RESULTS: Although the majority of mothers indicated the median silhouette as 'healthy', mothers of girls chose a fatter silhouette as 'healthy' more often than the mothers of boys (P = 0.001). By regression analysis, mothers' perception of a child's body shape was a significant determinant (P < 0.01) of blood pressure in both genders, and of anthropometric indices in girls only, independently of parental overweight, education, birth weight and physical activity. CONCLUSIONS: Our study suggests that maternal perception of a healthy body shape in children is associated with her child's blood pressure and degree of adiposity; with regard to the latter, differences between genders are apparent. PMID- 17917934 TI - Complex carbohydrate specificity of lectin from fruiting body of Ganoderma lucidum. A surface plasmon resonance study. AB - The thermodynamics and kinetics of binding of glycans and glycoproteins to Ganoderma lucidum lectin was studied using surface plasmon resonance. The lectin showed highest affinity for asialo triantennary N glycan (Ka = 3.52 x 10(5)) among the glycans tested. There was a several fold increase in affinity for glycoproteins compared to their corresponding glycans and coincident increase in contribution from enthalpy (DeltaH), suggesting the involvement of hydrogen bonding in the interaction as well as involvement of protein-protein interactions. Increased affinity also showed increase in unfavorable negative binding entropy (DeltaS) which was compensated with higher enthalpy. The glycoproteins showed faster association rates (k(1)) and the activation energy (E(1)) in the association process was much lower for the glycoproteins than glycans, resulting in their faster associations. These observations elaborate the role of protein matrix in lectin-glycoconjugate interaction. PMID- 17917936 TI - The skin is not lying. PMID- 17917935 TI - A randomized parallel study to assess the safety and efficacy of two different dosing regimens of 5% imiquimod in the treatment of superficial basal cell carcinoma. AB - OBJECTIVES: This study was designed to compare the safety and efficacy of two cycled dosing regimens of imiquimod 5% cream for treatment of superficial basal cell carcinoma (sBCC). METHODS: Patients (n = 32) were randomized to receive one of two treatment regimens: 8 weeks of treatment with once-daily dosing for alternate weeks (R1) and 5 weeks of once-daily dosing with a 1-week interval in the middle of the course (R2). Efficacy measures were tumour clearance at weeks 19 and 52 and measures of patients' acceptability. RESULTS: Data from 30 patients (13 females), 14 on R1 and 16 on R2, were analysed. The results revealed an initial clearance rate of 64% at week 19 for R1 and 81% for R2 (95% CI for difference: -14% to 45%, p = 0.21). However, clearance rates at week 52 were significantly different: 43% for R1 and 88% for R2 (95% CI for difference: 11% to 68%, p = 0.02). There was no difference in acceptability of treatment as measured by composite median visual analogue scores at week 8. CONCLUSION: Five weeks of 5% imiquimod cream once daily with a 1-week interval was more effective but as well tolerated as the 8-week alternate week regimen for sBCC. PMID- 17917937 TI - Outcomes in randomized controlled trials in psoriasis: what has changed over the last 20 years? AB - OBJECTIVE: In 1989, Marks et al. reviewed the different assessment tools employed in the evaluation of psoriasis severity before 1986. Our objective was to reproduce this seminal review to describe the changes over the last 20 years with regard to the evaluation of psoriasis severity in randomized clinical trials. DATA SOURCES: MeSH search on Pubmed for articles about psoriasis. STUDY SELECTION: Randomized clinical trials published in Archives of Dermatology, British Journal of Dermatology, Clinical and Experimental Dermatology and Journal of the American Academy of Dermatology from 1 January 2004 to 31 December 2005. DATA SYNTHESIS: Recent psoriasis trials implemented various clinical scoring systems in an attempt to assess psoriasis severity, but this is done without any apparent uniformity making direct comparisons between trials difficult. The predominant difference, which has occurred in the outcomes of psoriasis trials over the past 20 years, appears to be the introduction of an assessment of the patients' quality of life. CONCLUSION: The past 20 years have brought some improvements to the field of psoriasis evaluation but they also bring emphasis to the need for uniformity with regard to trial outcomes to ensure comparability and make better pooling of data from different randomized clinical trials possible for future meta-analyses. PMID- 17917938 TI - Is topical minoxidil solution effective on androgenetic alopecia in routine daily practice? AB - OBJECTIVES: Minoxidil solution stimulates hair growth in androgenetic alopecia. In order to maintain any beneficial effect, applications must continue indefinitely. The purpose of this study was to evaluate the ratio of patients who were satisfied with the drug and continued to use it versus those who were displeased and did not continue their treatment, and the reasons for the discontinuation. METHODS: A total of 1495 men aged 20-40 years who suffered from androgenetic hair loss were selected among patients who were referred to two private dermatologists. They were subjected to treatment with 5% topical minoxidil solution. These patients were treated with no difference from the routine office patients. Factors such as the duration of treatment, adverse effects, the patient's satisfaction and the causes of treatment cessation were also closely studied. RESULTS: Almost all the patients gradually avoided continuing the treatment. Only in a few patients was the cessation of medication due to adverse effects. The causes of discontinuation in the majority of patients were the low effect of medication and an aversion to this topical treatment method. CONCLUSIONS: The insignificant cosmetic effect of minoxidil solution caused discontinuity of treatment among almost all patients. PMID- 17917939 TI - DeWitt C. Baldwin Jr., MD: an interprofessional celebration. PMID- 17917940 TI - Interview with DeWitt C. Baldwin Jr. Interview by Lynda D'Avray. PMID- 17917941 TI - Some historical notes on interdisciplinary and interprofessional education and practice in health care in the USA. 1996. PMID- 17917942 TI - Maintenance of health care teams: internal and external dimensions. 1978. PMID- 17917943 TI - Interdisciplinary education and health team training: a model for learning and service. 1979. PMID- 17917944 TI - A model for recruitment and service--the University of Nevada's summer preceptorships in Indian communities. 1980. PMID- 17917945 TI - Interaction on health care teams. 1980. PMID- 17917946 TI - The British are coming: some observations on health care teams in Great Britain. 1982. PMID- 17917947 TI - Territoriality and power in the health professions. PMID- 17917948 TI - Collected works on interdisciplinary and interprofessional education and practice (listed chronologically by category). PMID- 17917949 TI - DeWitt C. Baldwin Jr., MD: an interprofessional celebration. PMID- 17917950 TI - Predicting worker exposure--the effect of ventilation velocity, free-stream turbulence and thermal condition. AB - Three-dimensional computational fluid dynamics (CFD) simulations were used to predict the flow field and resulting worker exposures when toxic airborne contaminants were released into the wake region of a mannequin that had its back to the airflow while holding the source of airborne contaminants. The effects of ventilation velocity, free-stream turbulence, and various thermal conditions on fluid flow and exposure levels were evaluated. The results showed good agreement between predicted and experimental concentrations at the mouth at a broad range of airflow velocities when the mannequin was both heated and unheated. When the mannequin was unheated, the exposure level decreased as the ventilation velocity increased. The expectation that buoyancy provided by the heat from the mannequin would be most important at very low velocities and decreasingly important at high velocities was proved true for both the predicted and observed exposures. The result was that when the mannequin was heated to normal human body temperatures, exposure levels had an inverted V relationship with velocity. These findings are important, since they call into question the common practice of modeling human exposures with mannequins at ambient temperatures. In addition, free-stream turbulence could be used to reduce worker exposure to airborne pollutants as suggested by the simulations. CFD enabled a detailed investigation of the effect of particular factors for exposure predictions in a cost-effective way. PMID- 17917951 TI - Field evaluation of an engineering control for respirable crystalline silica exposures during mortar removal. AB - During mortar removal with a right angle grinder, a building renovation process known as "tuck pointing," worker exposures to respirable crystalline silica can be as high as 5 mg/m(3), 100 times the recommended exposure limit developed by the National Institute for Occupational Safety and Health. To reduce the risk of silicosis among these workers, a vacuum cleaner can be used to exhaust 80 ft(3)/min (2.26 m(3)/min) from a hood mounted on the grinder. Field trials examined the ability of vacuum cleaners to maintain adequate exhaust ventilation rates and measure exposure outcomes when using this engineering control. These field trials involved task-based exposure measurement of respirable dust and crystalline silica exposures during mortar removal. These measurements were compared with published exposure data. Vacuum cleaner airflows were obtained by measuring and digitally logging vacuum cleaner static pressure at the inlet to the vacuum cleaner motor. Static pressures were converted to airflows based on experimentally determined fan curves. In two cases, video exposure monitoring was conducted to study the relationship between worker activities and dust exposure. Worker activities were video taped concurrent with aerosol photometer measurement of dust exposure and vacuum cleaner static pressure as a measure of airflow. During these field trials, respirable crystalline silica exposures for 22 samples had a geometric mean of 0.06 mg/m(3) and a range of less than 0.01 to 0.86 mg/m(3). For three other studies, respirable crystalline silica exposures during mortar removal have a geometric means of 1.1 to 0.35. Although this field study documented noticeably less exposure to crystalline silica, video exposure monitoring found that the local exhaust ventilation provided incomplete dust control due to low exhaust flow rates, certain work practices, and missing mortar. Vacuum cleaner airflow decrease had a range of 3 to 0.4 ft(3)/min (0.08 to 0.01 m(3)/sec(2)) over a range of vacuum cleaners, hose diameters, and hose lengths. To control worker exposure to respirable crystalline silica, local exhaust ventilation needs to be incorporated into a comprehensive silica control program that includes respiratory protection, worker training, and local exhaust ventilation. PMID- 17917952 TI - The stroke has only a minor influence on direction consistency in golf putting among elite players. AB - In the golf instructional literature, the putting stroke is typically given higher priority than green reading and aiming. The main purpose of this study was to assess the importance of the putting stroke for direction consistency in golf putting. Kinematic stroke parameters were recorded from 71 elite golf players (mean handicap = 1.8, s = 4.2) on 1301 putts from about 4 m. Of the different factors deciding stroke direction consistency, face angle was found to be the most important (80%), followed by putter path (17%) and impact point (3%). This suggests that improvements in consistency of putter path and impact point will have very little effect on overall putting direction consistency and should not be prioritized in the training of elite players. In addition, mean stroke direction variability for an elite player (European Tour) was found to be 0.39 degrees, which is good enough to hole about 95% of all 4-m putts. In practice, however, top professionals in tournaments only hole about 17% of 4-m putts. We conclude that the putting stroke of elite golfers has a relatively minor influence on direction consistency. PMID- 17917953 TI - Chronic lymphocytic leukaemia resistant to both fludarabine and alemtuzumab- double jeopardy. PMID- 17917954 TI - Mobilization regimens prior to stem-cell collection in patients with lymphoma: how to choose? PMID- 17917955 TI - Pulmonary arterial hypertension and myeloproliferative disorders. PMID- 17917956 TI - Plasma cell lysate as an antigen source in multiple myeloma immunotherapy. PMID- 17917957 TI - More progress defining the crosstalk between multiple myeloma and mesenchymal stem cells of the bone marrow. PMID- 17917958 TI - HMGA2 levels in CML: reflective of miRNA gene regulation in a hematopoietic tumor? PMID- 17917959 TI - Myelodysplastic syndromes, aging, and age: correlations, common mechanisms, and clinical implications. AB - Myelodysplastic syndromes (MDS) are a heterogenous group of myeloid neoplasms that develop primarily in elderly patients. Although a specific molecular basis for the predominant incidence of MDS in higher age groups remains unknown, several lines of evidence suggest that the biology of aging and the pathogenesis of MDS share several genetic, epigenetic, and molecular features. The current review attempts to delineate these common aspects as well as additional discriminative features that are specific for MDS and thus help explaining disease-evolution and progression. In addition, the present review discusses age as an important prognostic factor and co-variable to be considered in treatment algorithms in MDS. PMID- 17917960 TI - Use of bortezomib as induction therapy prior to stem cell transplantation in frontline treatment of multiple myeloma: impact on stem cell harvesting and engraftment. AB - High-dose melphalan therapy with peripheral blood stem cell (PBSC) transplantation is a standard treatment for younger patients with untreated multiple myeloma that results in high overall and complete response (CR) rates, and improved event-free and overall survival compared with standard chemotherapy alone. Induction therapy serves to reduce tumor burden prior to stem cell mobilization and thus must not adversely impact stem cell mobilization and harvesting, or engraftment following high dose therapy plus autologous stem cell transplantation. Bortezomib, an approved agent for the treatment of multiple myeloma patients who have received at least one prior therapy, is also being investigated in the frontline setting. Preclinical studies have demonstrated that bortezomib has no toxic effects on stem cells, megakaryocytes or neutrophil precursors, and causes only transient and reversible thrombocytopenia and neutropenia. Clinical studies with bortezomib-based induction regimens have demonstrated no adverse impact on PBSC harvest numbers nor on their quality as defined by engraftment times. These regimens appear to be well tolerated and highly active as induction therapy, with high response rates and consistently high CR rates. Randomized phase 3 studies comparing bortezomib-based regimens with current standard induction therapies are ongoing. PMID- 17917961 TI - The natural history of fludarabine-refractory chronic lymphocytic leukemia patients who fail alemtuzumab or have bulky lymphadenopathy. AB - The natural history and outcome of salvage treatment for patients with fludarabine-refractory chronic lymphocytic leukemia who are either refractory to alemtuzumab ("double-refractory") or ineligible for alemtuzumab due to bulky lymphadenopathy ("bulky fludarabine-refractory") have not been described. We present the outcomes of 99 such patients (double-refractory n = 58, bulky fludarabine-refractory n = 41) undergoing their first salvage treatment at our center. Patients received a variety of salvage regimens including monoclonal antibodies (n = 15), single-agent cytotoxic drugs (n = 14), purine analogue combination regimens (n = 21), intensive combination chemotherapy (n = 36), allogeneic stem cell transplantation (SCT; n = 4), or other therapies (n = 9). Overall response to first salvage therapy other than SCT was 23%, with no complete responses. All four patients who underwent SCT as first salvage achieved complete remission. Early death (within 8 weeks of commencing first salvage) occurred in 13% of patients, and 54% of patients experienced a major infection during therapy. Overall survival was 9 months, with hemoglobin < 11 g/dL (hazard ratio 2.3), hepatomegaly (hazard ratio 2.4), and performance status > or = 2 (hazard ratio 1.9) being significant independent predictors of inferior survival. PMID- 17917962 TI - Antiangiogenic activity of thalidomide in combination with fludarabine, carboplatin, and topotecan for high-risk acute myelogenous leukemia. AB - Forty-two patients with poor prognosis AML were enrolled in a phase II study combining fludarabine, carboplatin, and topotecan (FCT) with thalidomide. Laboratory correlates included serum vascular endothelial growth factor levels (VEGF) and bone marrow microvascular density (MVD). Ten of 42 (24%) patients achieved a complete remission (CR or CRp). Serious thrombotic adverse events were observed in 5 patients suggesting that the combination of cytotoxic chemotherapy and thalidomide may be thrombogenic despite significant thrombocytopenia. VEGF did not correlate with response to therapy, while a trend towards decreased MVD was noted in patients who achieved CR. The addition of thalidomide did not significantly influence angiogenic markers. It is not clear that thalidomide adds any efficacy to the FCT regimen. PMID- 17917963 TI - Intermediate dose etoposide plus G-CSF 16 g/kg is more effective than cyclophosphamide 4 g/m(2) plus G-CSF 10 g/kg in PBSC mobilization of lymphoma patients. AB - We designed intermediate dose etoposide + G-CSF 16 microg/kg as a Peripheral Blood Stem Cell (PBSC) mobilization schedule suitable for outpatient administration. Forty-one Lymphoma patients received intermediate dose etoposide (200 mg/m(2) i.v. day +1, +2, +3) +G-CSF 16 microg/kg/day. Results of PBSC mobilization in these patients were compared with those of a group of 37 lymphoma patients mobilized using cyclophosphamide (CTX) at dosage of 4 g/m(2) + G-CSF 10 microg/kg/die. Mean peak of CD34+ cells achieved in P.B. and total CD34+ cells harvested were higher in patients mobilized with intermediate dose etoposide (p = 0.003 and p = 0.004, respectively). After transplantation recovery of polymorphonucleate neutrophils (PMN) > 0.5 x 10(9)/L did not differ significantly between groups: 11.7 days in intermediate dose etoposide group and 11.5 days in CTX group (p = 0.7). Intermediate dose etoposide + G-CSF 16 microg/kg resulted in a maximum length of neutropenia (PMN < 0.5 x 10(9)/L) of 2 days and neutropenic fever was registered during only 3/41 courses (7.3%). Intermediate dose etoposide + G-CSF 16 microg/kg is a highly effective mobilizing therapy, further, it has the advantage of low hematologic toxicity and can be easily administered as outpatient treatment. PMID- 17917964 TI - The pretransplant Follicular Lymphoma International Prognostic Index is associated with survival of follicular lymphoma patients undergoing autologous hematopoietic stem cell transplantation. AB - We retrospectively evaluated the association of the Follicular Lymphoma International Prognostic Index (FLIPI) and other characteristics with survival following high-dose therapy and autologous stem cell transplantation (ASCT) in 207 consecutive follicular lymphoma (FL) patients. The FLIPI was associated with OS both when evaluated as a categorical variable (0 - 1 vs. 2 vs. 3 vs. 4, p = 0.01, global test) and a continuous linear variable (p = 0.002). The association of FLIPI with survival appeared to be more relevant for patients who received standard conditioning regimens compared to those that were treated with high-dose radioimmunotherapy (p = 0.004). Among all patients, mortality was also associated with chemosensitive disease (HR = 0.47, p = 0.01) or untreated relapse (HR = 0.20, p = 0.0002) vs. chemoresistant disease, and > or =2 extranodal sites (HR = 2.82, p = 0.03) after adjusting for FLIPI. These data suggest that the FLIPI and select non-FLIPI factors after adjustment for the FLIPI are associated with survival in FL patients undergoing ASCT. PMID- 17917966 TI - Clinical outcome of incidental pelvic node malignant B-cell lymphomas discovered at the time of radical prostatectomy. AB - Incidental pelvic node malignant B-cell lymphomas diagnosed at the time of radical prostatectomy are rare. Their clinical outcome has not been studied. We studied thirteen such cases with long-term clinical follow-up. Patients were followed between 9 and 94 months after surgery. Of 13 cases, 9 were chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), 3 marginal zone B-cell lymphoma (MZL) and 1 mantle cell lymphoma (MCL). All 13 patients did not receive radiation or chemotherapy; and five of 13 cases showed hematologic evidence of lymphoma progression between 1 and 5 months after radical prostatectomy. After progression, the mantle cell lymphoma patient received aggressive chemotherapy and had systemic dissemination. Two of 13 cases had recurrent prostate carcinoma. None of 13 patients had died from lymphoma or prostate carcinoma at the last follow-up. In conclusion, most incidental pelvic node lymphomas (8/13) showed no evidence of systemic dissemination to peripheral blood or bone marrow after a mean 42.8 weeks of follow-up despite the fact that no additional treatment was given. Strong consideration should be given to withholding further treatment in patients diagnosed with pelvic low-grade B-cell lymphoma at the time of radical prostatectomy until disease progression occurs. PMID- 17917965 TI - Autologous stem cell transplantation for patients with active Hodgkin's lymphoma: long-term outcome of 61 patients from a single institution. AB - Sixty-one patients with refractory or relapsed Hodgkin's lymphoma (HL) underwent high-dose chemotherapy and autologous stem cell transplantation (ASCT). All patients had active HL at the time of ASCT: 13 patients had partial remission, 14 refractory disease, 18 sensitive relapse, 4 resistant relapse, and 12 nontreated relapse. Overall transplant-related mortality (TRM) was 16.4% at 1 year. Twenty eight patients (46%) achieved complete remission (CR). Actuarial 5-year overall survival (OS) and progression-free survival (PFS) were 51% and 47%, respectively. Patients with positive gallium-67 scintigraphy at 3 - 6 months after transplantation had a worse PFS at 5 years (28%) than those with negative 67Ga scan (80%) (p = 0.016), whereas no statistical differences were observed between patients with residual mass and those in CR according to computed tomography scan. In multivariate analysis, bulky disease at diagnosis, bone marrow stem cells, and stage IV at transplant were the only adverse prognostic factors significantly influencing OS. Bulky disease at diagnosis and stage IV at transplant adversely influenced PFS. Although long-term outcome of patients with active HL at the time of ASCT is poor due to a high TRM and a low CR after transplantation, a subgroup of patients with no adverse prognostic factors at ASCT gain benefit from this treatment. PMID- 17917967 TI - Myeloid leukemias express a broad spectrum of VEGF receptors including neuropilin 1 (NRP-1) and NRP-2. AB - Vascular endothelial growth factor (VEGF) is produced in neoplastic cells in various myeloid neoplasms and may act as an autocrine growth-regulator. We have examined the expression of five VEGF receptors (VEGR1/Flt-1, VEGFR2/KDR, Flt-4, neuropilin-1 = NRP-1, NRP-2) in leukemic cells obtained from patients with acute myeloid leukemia (n = 28), chronic myeloid leukemia (n = 14), chronic eosinophilic leukemia (n = 3), chronic myelomonocytic leukemia (n = 9), or mast cell leukemia/systemic mastocytosis (n = 3) as well as in respective cell lines. Expression of VEGFR mRNA was analyzed by RT-PCR, and expression of VEGFR protein by immunocytochemistry. In most patients, leukemic cells expressed NRP-1 mRNA and NRP-2 mRNA independent of the type of disease. By contrast, transcripts for Flt 1, KDR, and Flt-4 were expressed variably without a clear correlation to the type of leukemia. Expression of VEGF receptors was also demonstrable at the protein level in all cases tested. In conclusion, neoplastic cells in myeloid leukemias frequently express VEGFR including NRP-1 and NRP-2. PMID- 17917968 TI - Quantitative expression analysis in peripheral blood of patients with chronic myeloid leukaemia: correlation between HMGA2 expression and white blood cell count. AB - The architectural transcription factor HMGA2 is highly expressed during embryogenesis but scarcely detectable in non-dividing adult cells. Previously, HMGA2 re-expression was detected in blood from CML patients by conventional RT PCR, while blood samples from healthy volunteers were HMGA2 negative. Using the sensitive method of real-time quantitative RT-PCR, herein HMGA2 expression was detectable not only in peripheral blood from leukaemia patients but also in blood from healthy donors. Statistical analysis revealed a highly significant correlation between white blood cell count and HMGA2 transcript levels. The results indicate that up-regulation of HMGA2 expression is correlated to the undifferentiated phenotype of leukaemic cells accumulating during progression of chronic phase to blast crisis. PMID- 17917969 TI - VEGF-D is expressed in activated lymphoid cells and in tumors of hematopoietic and lymphoid tissues. AB - Vascular Endothelial Growth Factor (VEGF)-D is a member of the VEGF family of angiogenic growth factors that activate the Vascular Endothelial Growth Factor Receptor (VEGFR)-2 and VEGFR-3, which are mainly expressed in blood and lymphatic vessels. Here we have analyzed by using monoclonal antibodies, the expression of VEGF-D and its cognate receptor VEGFR-3 in normal and pathologic bone marrow and lymph node biopsies. This analysis revealed that VEGF-D is expressed in B cells of the germinal centers, scattered B and T blasts, myeloid progenitors, acute leukemia, several types of non Hodgkin lymphoma, and classical Hodgkin's lymphoma. In normal tissues VEGFR-3 was only expressed in fenestrated capillaries of bone marrow and in lymphatic vessels of lymph nodes, while in VEGF-D expressing tumors newly formed vessels, but not malignant cells, showed high VEGFR-3 expression. These data suggest that VEGF-D could contribute to leukemia and lymphoma growth via the induction of angiogenesis in bone marrow and lymphoid tissues. PMID- 17917970 TI - Induction of multiple myeloma-specific cytotoxic T lymphocyte stimulation by dendritic cell pulsing with purified and optimized myeloma cell lysates. AB - We investigated the possibility of immunotherapy for multiple myeloma (MM) using myeloma-specific cytotoxic T lymphocytes (CTLs) that were stimulated in vitro by dendritic cells (DCs) pulsing with purified and optimized myeloma lysates. CD14(+) cells were cultured in the presence of GM-CSF and IL-4. On day 6, the immature DCs were pulsed with the purified myeloma cell lysates, and then maturation of the DCs was induced by the addition of a cytokine cocktail. There were no differences in the phenotypic expressions of mature DCs that were generated by pulsing with CD138(+) cell lysates or total cell lysates. In optimization of the concentration of myeloma lysates, DCs pulsed with 10 microg/mL of myeloma lysate had greater allogeneic T-cell stimulatory capacities than those pulsed with higher concentrations of myeloma lysates. The CTL lines generated by DCs pulsed with myeloma lysates demonstrated potent cytotoxic activities against autologous target cells, but not against HLA-A2(-) cell lines or K562 cell lines. The DCs pulsed with myeloma lysates demonstrated a higher stimulatory capacity for autologous CTL compared with mature nonpulsed DCs. These results suggested that the DCs pulsed with purified and optimized myeloma cell lysates could generate potent myeloma-specific CTLs for approaches in MM. PMID- 17917971 TI - Mesenchymal stem cell abnormalities in patients with multiple myeloma. AB - Osteolytic bone lesions are common in patients with multiple myeloma (MM), a clonal plasma cell disorder, and result from increased osteoclastic bone resorption and decreased osteoblastic bone formation. Because mesenchymal stem cells (MSCs) are committed towards cells of the osteoblast lineage, we compared the in vitro characteristics of MSCs from the bone marrow of 18 MM patients (MM MSCs) and eight normal donors (ND-MSCs). MM-MSCs displayed deficient growth that could be explained in part by the reduced expression of several growth factor receptors on the surface of MM-MSCs compared with ND-MSCs. Receptor downregulation was observed on RT-PCR analysis. A major finding was an approximately fivefold higher expression of osteoblast inhibitor DKK1 at transcript and protein levels in MM-MSCs than ND-MSCs. These data suggest that defective osteoblast function in patients with advanced MM may be related not only to factors released by tumor myeloma cells but also to MSC abnormalities. PMID- 17917972 TI - Three-dimensional culturing of the Hodgkin lymphoma cell-line L1236 induces a HL tissue-like gene expression pattern. AB - To overcome some limitations of in vitro established cell-line tumor models for Hodgkin lymphoma (HL), we explored whether culturing in a three-dimensional (3D) matrix could improve the quality of the model. We used a novel designer peptide based self-organizing matrix. The gene expression profile of the 3D-cultured HL derived cell-line L1236 was compared with that of suspension-cultured (2D) L1236, as well as to the gene expression profile found in HL tumor samples. To validate our results we also included a gene expression data set of laser captured Hodgkin Reed - Sternberg (H-RS) cells. The gene expression profiles were analyzed using Affymetrix technology. We found that the 3D culture affected gene expression of a HL derived cell-line inducing a more tumor-related expression profile. 3D culture affected the expression of 500 genes in L1236, upregulating genes involved in immune response and apoptosis and downregulating genes involved in cell division. It also affected genes involved in actin filament polymerization. PMID- 17917973 TI - Dasatinib restores full donor chimerism in a patient with imatinib-resistant Ph+ ALL relapsing after unrelated cord blood transplantation. PMID- 17917974 TI - Response to highly active antiretroviral therapy as the only therapy in an HIV infected patient with interfollicular Hodgkin's lymphoma. PMID- 17917975 TI - Immunoglobulin receptor evolution in follicular lymphoma and a review of literature. PMID- 17917977 TI - Amyloid-associated amyloidosis in a HCV carrier with non-Hodgkin's lymphoma who had been treated with autologous stem cell transplantation and rituximab. PMID- 17917976 TI - Angioimmunoblastic T-cell lymphoma with an evolving Epstein Barr virus-positive diffuse large B-cell lymphoma with unusual clinical and pathologic findings. PMID- 17917978 TI - Blastoid mantle cell lymphoma with t(2;8) (p12;q24). PMID- 17917979 TI - Unusual profiles of pediatric acute lymphoblastic leukemia with MLL gene rearrangement. PMID- 17917980 TI - High-speed registration of phonation-related glottal area variation during artificial lengthening of the vocal tract. AB - Vocal exercises that increase the vocal tract impedance are widely used in voice training and therapy. The present study applies a versatile methodology to investigate phonation during varying artificial extension of the vocal tract. Two males and one female phonated into a hard-walled plastic tube (phi 2 cm), whose physical length was randomly pair-wise changed between 30 cm, 60 cm and 100 cm. High-speed image (1900 f/sec) sequences of the vocal folds were obtained via a rigid endoscope. Acoustic and electroglottographic signals (EGG) were recorded. Oral pressure during shuttering of the tube was used to give an estimate of subglottic pressure (Psub). The only trend observed was that with the two longer tubes compared to the shortest one, fundamental frequency was lower, open time of the glottis shorter, and Psub higher. The results may partly reflect increased vocal tract impedance as such and partly the increased vocal effort to compensate for it. In other parameters there were individual differences in tube length related changes, suggesting complexity of the coupling between supraglottic space and the glottis. PMID- 17917981 TI - Can vocal economy in phonation be increased with an artificially lengthened vocal tract? A computer modeling study. AB - Voiced obstruents and phonation into tubes are widely used as vocal exercises. They increase the inertive reactance of the vocal tract in the 200-1000 Hz range and thereby reinforce vocal fold vibration. But the effect is strong only when the epilarynx tube is also narrowed. The present study focused on the effects of a 'resonance tube' (27 cm in length, 0.5 cm2 cross-sectional area, hard walls) on vocal tract reactance and the accompanying economy of voice production (defined as maximum flow declination rate (MFDR), divided by maximum area declination rate (MADR)). The vowel /u/ and phonation into the tube were simulated with a computer model. Three values were given to the cross-sectional area of the epilarynx tube (0.2 cm2, 0.5 cm2, and 1.6 cm2), which is at the opposite end of the vocal tract from the artificial 'resonance tube'. The degree of glottal adduction was varied in order to find the economy maximum for each epilarynx tube setting. Results showed that the 'resonance tube' lowered F1 from 300 Hz to 150 Hz and doubled the vocal tract inertive reactance at F0=100 Hz. The largest economy with the 'resonance tube' was obtained when the epilarynx tube was narrowed (relative to the rest of the vocal tract) and sufficiently tight adduction was used. Most importantly, the intraoral acoustic pressure (calculated at 0.8 cm behind the lips) was tripled with the tube. The results suggest that by optimizing the vibratory sensations in the face that are attributed to increased intraoral acoustic pressure, phonation into a tube may assist a trainee in finding an optimal glottal and epilaryngeal setting for the greatest vocal economy. PMID- 17917982 TI - Communicative satisfaction of male-to-female transsexuals. AB - The aim of this qualitative study was to gain an understanding of the way male-to female transsexuals perceived their communication and their satisfaction with it in different aspects of their lives. Three focus groups were conducted for the purposes of this study, each consisting of four participants. The transcribed dialogues of the groups were analysed using a grounded theory approach to find the common underlying themes. These then formed the basis of the five major categories that identified key themes relating to transsexuals' communicative satisfaction: communicative situations, emotions, groups of people, other contributing factors, and features of communication. The results revealed that although participants commented on the situations and contexts in which they experienced decreased communicative satisfaction, it was not always important to them to maintain their female communicative patterns in situations in which they felt comfortable. These five categories guided the development of a two-part pilot Functional Communicative Satisfaction Questionnaire (FCSQ). The FCSQ explores a variety of situations that the participants identified as impacting on their communicative satisfaction. Further development of the FCSQ is likely to enable speech pathologists to have a more informed approach to the management of this population. PMID- 17917983 TI - Establishing health promotion in the modern medical curriculum: a case study. AB - BACKGROUND: The subject of health promotion has yet to be established as core content in undergraduate medical curricula, despite calls from the medical education community and from regulatory bodies such as the UK's General Medical Council more than a decade ago. At the same time, concerns about the decline in popularity of teaching public health have been raised. AIMS: The challenges associated with teaching both disciplines--health promotion and public health- are complex and linked, but, for health promotion, we argue that two factors are paramount: the nature of the discipline, and how it is presented as being relevant to undergraduate medical curricula, in particular those with integrated courses. METHODS: Using the approach taken at one medical school as a case study, this paper discusses the factors surrounding health-promotion curricular development. It explores these challenges, including the relevance of health promotion to medical students, medical teachers, and medical practitioners. RESULTS: These developments are linked to process and content and, therefore, to learning outcomes. CONCLUSIONS: There is a paucity of information from medical schools about their approaches to health promotion curricula content and teaching. This paper may provide schools with ideas for the development of their own health promotion content. PMID- 17917984 TI - Validity, reliability, feasibility and satisfaction of the Mini-Clinical Evaluation Exercise (Mini-CEX) for cardiology residency training. AB - AIMS: The purpose of the study was to determine the validity, reliability, feasibility and satisfaction of the Mini-CEX. METHODS AND RESULTS: From May 2003 to December 2004, 108 residents from 17 cardiology residency programs in Buenos Aires were monitored by the educational board of the Argentine Society of Cardiology. Validity was evaluated by the instrument's capability to discriminate between pre-existing levels of clinical seniority. For reliability, generalisability theory was used. Feasibility was defined by a minimum number of completed observations: 50% of the residents obtaining at least four Mini-CEX's. Satisfaction was evaluated through a one to nine rating scale from the evaluators, and residents' perspectives. The total number of encounters was 253. Regarding validity, Mini-CEX was able to discriminate significantly between residents of different seniority. Reliability analysis indicated that a minimum of ten evaluations are necessary to produce a minimally reliable inference, but more are preferable. Feasibility was poor: 15% of the residents were evaluated four or more times during the study period. High satisfaction ratings from evaluators' and residents' were achieved. CONCLUSION: Mini-CEX discriminates between pre-existing levels of seniority, requires considerable sampling to achieve sufficient reliability, and was not feasible within the current circumstances, but it was considered a valuable assessment tool as indicated by the evaluators' and residents' satisfaction ratings. PMID- 17917985 TI - Collaboration of junior students and residents in a teacher course for senior medical students. AB - BACKGROUND: This paper describes the curriculum and impact of an innovative resident-as-teacher course at the University of Massachusetts Medical School. The intent of the course is to prepare students across undergraduate and graduate medical education for their roles as teacher, learner and leader during residency. METHOD: The elective introduces teaching skills curricula during year two of medical school and integrates it into residency years. The course was developed with the premise that integrating facets of graduate and undergraduate medical education offers students diverse perspectives of the teaching/learning process. Students completed a pre-/post-course questionnaire and an end-of-course evaluation, from which data were gathered to determine course impact. RESULTS: Students reported increased confidence levels in their teaching skills from pre- to post-test. Post-test data indicated that 88% of students 'agreed' and 12% 'strongly agreed' that the course equipped them with 'skills which will enable [them] to provide teaching that supports effective learning', an increase from pre-test data which indicated that 50% 'agreed' and 50% 'disagreed'. CONCLUSION: Results indicate the course did effect a change in students' attitudes to teaching as evident from comments such as, 'I will be a better teacher because I have been given the appropriate tools'. PMID- 17917986 TI - A teaching rotation and a student teaching qualification for senior medical students. AB - BACKGROUND: Physicians have a scholarly role, including educating others, be it patients, colleagues or students. This led UMC Utrecht to organize a one-week obligatory teacher training course for senior medical students and a six-week elective teaching rotation. Students who complete both may receive a student teaching qualification. They must show didactic skill, have experience with teaching and learn about medical education topics. METHOD: Between October 2004 and December 2006 60 students (about 10% of all final-year students) took the six week rotation. Rotating students are attached to a 4-6 week undergraduate curriculum unit. The rotation includes 30 hours of independent teaching of junior medical students, studying the BMJ 's ABC of Learning and Teaching series and sitting a written exam on it, constructing 14 test questions, completing an advisory project for the unit coordinator and writing an essay on a medical education topic of choice. CONCLUSIONS: The senior students appear to attain the objectives of the rotation and evaluate it positively. The coordinating teacher supervisors and the younger students are all generally satisfied to very satisfied with the student teachers. The curriculum benefits from the advisory projects and most essays are so well written that a selection is published as a booklet presented for teachers. These graduates start their career with a teaching qualification. PMID- 17917988 TI - Educators' perspective of online course design and delivery. AB - BACKGROUND: With advances in technology, educators are increasingly involved in the delivery of online courses. This paper reports a qualitative descriptive study from the educators' perspective of developing and delivering a fully online course to undergraduate health science students. METHOD: The authors engaged in reflective conversations about the course which were audio-taped, and the resultant transcripts analysed thematically. RESULTS: Two main themes emerged: (1) Arousing students' interest, which represented the central motivator in the formulation and implementation of the course, and (2) Working with larger numbers of students, which highlighted the most significant challenge encountered during course delivery. There are considerable workload issues for courses which attempt to use material that is relevant, applied to students' interests, requires collaboration, and includes learning activities linked to sequential assessment. Time must be allowed for consistent, frequent and focused engagement with the students online in order to achieve a quality teaching and learning environment. CONCLUSIONS: The findings revealed that ongoing tension exists between aspiring to provide high quality, pedagogically sound, interesting and collaborative online learning opportunities and the workload and time demand realities of enacting these intentions. PMID- 17917989 TI - Educational environment in intensive care medicine--use of Postgraduate Hospital Educational Environment Measure (PHEEM). AB - BACKGROUND: The educational climate is an important measure within medical education. This is because there are still accounts of poor teaching, humiliation, bullying and harassment of doctors in training. Deaneries and schools must be able to demonstrate to the Postgraduate Medical Education and Training Board that trainees are working and learning in a good environment. METHODS: This study used the Postgraduate Hospital Educational Environment Measure (PHEEM) to measure the educational climate in nine intensive care training schemes within hospitals in England and Scotland. 134 trainees replied out of 190 (71% response). Neither the identities of the nine units nor the trainees were known to the researchers. RESULTS: The results showed that there was a good overall educational climate in the intensive care units studied, with no racism or sexism, and trainees were happy with their teaching, their support and the work they did. The junior trainees (house officers and senior house officers) perceived a significantly better climate than did the senior trainees (specialist registrars). There were also significant differences in scores for the nine different intensive care units. CONCLUSIONS: PHEEM has proved to be a reliable and consistent tool to assess educational climate with an overall Cronbach's alpha of 0.921. PMID- 17917990 TI - PBL curriculum improves medical students' participation in small-group tutorials. AB - BACKGROUND: Group learning is the core of problem-based learning (PBL) but has not been extensively studied, especially in Asian students. METHODS: This study compared students of PBL and non-PBL curricula in students' talking time and participation in small-group tutorials in a medical school in Asia. The proportions of student talking of 46 tutorials in three teaching rotations of the PBL curriculum and those of 43 corresponding tutorials in the non-PBL curriculum were counted. Twelve videotapes of tutorials (six from each curriculum), stratified for tutor, case scenario and students' learning stage, were randomly selected and transcribed. They were rated with the group-interaction (5 items) and active-participation (four items) tutorial assessment scales developed by Valle et al. These outcomes were compared between the students of PBL and non-PBL curricula. RESULTS: Students from the PBL curriculum talked significantly more. In only two (4.7%) of 43 tutorials in the non-PBL curriculum did the students talk more than the tutors; but students talked more than the tutors in 17 (37.0%) of 46 tutorials in the PBL curriculum. PBL students scored significantly higher than non-PBL students in all items except one item (respect to peers) of the tutorial assessment scales, and in the mean scores of both the group interaction scale (items 1-5) and the active participation scale (items 6-9). CONCLUSIONS: The results suggested that PBL starting from the early years of a medical curriculum was associated with more active student participation, interaction and collaboration in small-group tutorials. PMID- 17917991 TI - The development and validation of the mini-surgical theatre educational environment measure. AB - BACKGROUND: The operating theatre educational environment during medical school may influence student's perceptions of a surgical career. Interest in a surgical career is declining both in the USA and the UK. This study sought to develop and validate a practical questionnaire for distribution to medical undergraduates to assess their operating theatre educational environment. METHODS: A previously published questionnaire--the Surgical Theatre Educational Environment Measure (STEEM) (validated on postgraduates) was distributed to 83 final year medical students. The STEEM was found to be a reliable and valid tool for use on undergraduates (Cronbach's alpha = 0.86). From this preliminary study, factor analysis identified 13 factors covering 73% of the variance, which enabled the development of a shorter 14-item questionnaire (mini-STEEM). RESULTS: The mini STEEM was distributed to 99 medical students following their final year surgical placement at a UK medical school, with a 100% response rate. The mini-STEEM was shown to be a valid, reliable (Cronbach's alpha = 0.80) and practical tool, which was easy to complete, code and analyse. CONCLUSIONS: The results showed that the overall climate within the operating theatre for medical undergraduates in one UK Medical School was good with no gender differences in the overall scores. PMID- 17917992 TI - Improving general practitioners' interviewing skills in managing patients with depression and anxiety: a randomized controlled clinical trial. AB - BACKGROUND: Studies regarding the effectiveness of CME programmes on physicians' behaviour and communication skills showed inconsistent results. Few randomized controlled trials have been conducted in Asia. METHODS: To evaluate the effectiveness of a 4 2-hour education programme to improve GP interviewing behaviours, 16 general practitioners were randomized to the intervention and control groups, respectively. Physicians assigned to the intervention group received 8 hours of training emphasizing interviewing behaviours in the diagnosis and treatment of depression and generalized anxiety disorders (GDS). Those assigned to the control group did not receive any training until the completion of study. Standardized patients were used to evaluate the performance of physicians. Two consultations before and after enrolling in the education programme were videotaped. Independent evaluations of consultations were made by a trained clinical psychologist and a social worker blinded to the study status of physicians. The rating schedule for the videotapes was based on the tasks listed on the Calgary Cambridge Observation Guide. RESULTS: The change of score between the intervention and control physicians was significantly different in 'active listening and facilitating patients' response' (p = 0.011) with the intervention physicians having improvement of score. For 'non-verbals', 'understanding patient's perspective' and 'negotiating mutual plan of action', positive change of score in the intervention physicians were seen when compared to that of the control, although the difference did not reach statistical significance (p = 0.06, p = 0.05, p = 0.06, respectively). However, for 'opening', 'structuring the consultation', 'explanation and planning' and 'closure', there were no statistical significant differences between control and intervention group. CONCLUSIONS: Our results showed that only certain communication skills, such as active listening and facilitating patient's response, can be taught in the management of depression and generalized anxiety disorder (GAD) in Chinese primary care physicians. PMID- 17917995 TI - Tonatiuh II: assisting manipulator for laparoscopic surgery. AB - In this article we show the design of the Tonatiuh II robotic manipulator. This robotic assistant has an original electromechanical configuration and respects the laparoscope center of insertion as an invariant point for navigation in the work space. The manipulator went through several stages before reaching its final version. Surgical trials have shown the robot to be useful in the operating room and as a training assistant in specialty microsurgery. PMID- 17917996 TI - Laparoscopic splenectomy plus cholecystectomy for treating hereditary spherocytosis combined with cholelithiasis in siblings. AB - With the advancements that have been made in the field of minimally invasive surgery, combined laparoscopic procedures are now being performed for treating coexisting abdominal pathologies during the same surgery; this combined surgery thereby offers the maximum benefits to the patients. In this paper we describe how we perform laparoscopic splenectomy plus cholecystectomy at the same time to treat hereditary spherocytosis combined with cholelithiasis in two siblings. PMID- 17917997 TI - Diabetes, revascularization and CABg: when there is smoke, there is a fire! PMID- 17917998 TI - Expression of CD69 on T-cell subsets in HIV-1 disease. AB - CD69 is the earliest activation marker newly synthesized and expressed during T lymphocyte activation. In this study, a whole-blood flow-cytometry-based assay was used to assess expression of the activation antigen CD69 on CD4 and CD8 T lymphocytes, and the co-expression of CD69 and CD28 on T cells. The expression of CD69 was studied in both unstimulated and in phytohaemagglutinin (PHA)- or anti CD(3)/CD(28)-stimulated, 4-h culture, samples. The production of IL-2, IFN-gamma or both cytokines, in CD69(+) T cells, in response to Staphylococcus enterotoxin B was also tested. Fifty-three HIV-1-infected and 21 healthy volunteers participated in this study. In both PHA- and anti-CD(3)/CD(28)-stimulated cultures the percentage of CD69 on CD3(+)CD4(+) T cells was significantly lower in AIDS (and non-responders to HAART) versus healthy controls and the other HIV 1(+) groups. A decrease of CD69(+)CD28(+) T cells after PHA or MoAbs stimulation is noticed in AIDS. No difference in cytokine production was noticed between healthy volunteers and HIV-1(+) patients. Our results suggest that the expression of CD69 is affected only in the AIDS stage and in the non-responders to HAART patients. PMID- 17917999 TI - Regression of lactose malabsorption in coeliac patients after receiving a gluten free diet. AB - OBJECTIVE: In a recent study by our group, it was shown that a large proportion of patients with lactose malabsorption and with no bacterial overgrowth are affected by silent coeliac disease (CD). Our aim was to evaluate the effect of a gluten-free diet on lactose malabsorption assessed using the hydrogen lactose breath test (LBT) and also the relationship with normalization of duodenal biopsies in coeliac patients. MATERIAL AND METHODS: Fifteen patients (11 F, 4 M; mean age 35.8+/-6) affected by CD with a positive LBT and negative glucose breath test were enrolled. All were started on a gluten-free diet and were re-evaluated after 6 months by LBT and after 12 months by both LBT and upper gastrointestinal endoscopy with biopsies. RESULTS: LBT normalization was observed in 1 out of 15 patients (6.7%) after 6 months and in 9 of the remaining 14 (64.2%) after 12 months. Duodenal biopsies showed normal villi in 8 patients, partial villous atrophy in 5 and total atrophy in 2. CONCLUSIONS: The present study shows that a large proportion of CD patients experience a regression of lactose malabsorption after receiving a gluten-free diet. This may be related to normalization of the brush border with an improvement of lactase enzyme activity. LBT should be performed after 12 months in CD patients on a gluten-free diet in order to assess the persistence/disappearance of lactose malabsorption, thus avoiding an unnecessary lactose-free diet. PMID- 17918000 TI - Concept of Crohn's disease being conditioned by four main components, and irritable bowel syndrome being an incomplete Crohn's disease. AB - Several mechanisms have been proposed for the development of Crohn's disease. Evidence in favour of a unifying 4-component concept to explain the development of Crohn's disease is presented. The four components are a genetic predisposition to an increased intestinal permeability, the key and initial triggering factor being an oral-pharyngeal bacterium that increases the mucosal permeability of the small intestine with only a minimal inflammatory reaction, an adherent-invasive strain of Escherichia coli that penetrates the mucosa and causes an acute inflammatory reaction in the intestinal wall, and finally a secondary invasion of bacteria causing the chronic inflammatory characteristics. Irritable bowel syndrome (IBS) is a functional bowel disorder with intermittent symptoms of varying intensity. Clinically, there is evidence to suggest a link between IBS patients with diarrhoea and patients with Crohn's disease. The common denominator and initial trigger for IBS with diarrhoea and Crohn's disease seems to be an increased small intestinal permeability, probably caused by an oral-pharyngeal bacterial strain. The important missing factor in IBS patients seems to be the adherent-invasive strain of E. coli in the proximal colon, causing the acute inflammatory process in patients with Crohn's disease. IBS with diarrhoea can then be looked upon as an incomplete Crohn's disease. PMID- 17918001 TI - Location of liver metastases reflects the site of the primary colorectal carcinoma. AB - OBJECTIVE: The present study was designed to investigate whether the different venous return of different locations of colorectal carcinomas affects the lobar distribution of metastases to the liver, due to the "streaming" within the portal vein. MATERIAL AND METHODS: The site of the primary colorectal carcinoma was divided into the right- and left hemicolon according to the different venous drainage via the superior and the inferior mesenteric/splenic vein. Both groups were analyzed for the distribution of the metastases in the liver. The anatomic site of the liver metastases was detected by intraoperative exploration and differentiated between the two lobes using the Cantlie line. RESULTS: Out of a total of 178 patients, 109 men and 69 women with 264 metastases were eligible for the study. The ratio of metastases in the right and left hemiliver was 3.6:1 for 35 right-sided primary tumors (p=0.002) compared with 2.1:1 for 143 left-sided primary tumors (p=NS). No significant differences were evident for the sub analysis of involved liver segments. CONCLUSIONS: The results of our study support the existence of the "streaming" effect in the portal vein. Right-sided colon carcinomas predominantly involve the right hemiliver, while left-sided colon carcinomas involve the liver homogeneously, considering the size ratio of the right to left liver lobe, which is about 2:1. Knowledge of streaming may help us to understand the spread of abdominal malignancies and may provide a reference concerning the possible primary site depending on metastatic distribution in the liver. PMID- 17918002 TI - Endoscopy and magnetic resonance imaging in patients with Crohn's disease: a retrospective single-centre comparative study. AB - OBJECTIVE: There is ongoing debate about which imaging modality is best for patients with inflammatory bowel diseases. Magnetic resonance imaging (MRI) has been successfully used to evaluate the jejunum and the ileum. Because virtual colonoscopy by MRI requires bowel cleansing and/or rectal filling, endoscopy is preferred for assessment of the colon. However, hydro-MRI without special bowel preparation may be sufficient as a diagnostic tool if specifically targeted in the course of a known disease. The aim of this study was retrospectively to assess the correlation of endoscopy, histology and MRI findings for the terminal ileum and the colon in a cohort of patients with Crohn's disease. MATERIAL AND METHODS: In all, 60 patients with a confirmed diagnosis of Crohn's disease were included in the study. Here, 412 anatomical segments of the colon were analysed by MRI, 401 by endoscopy and 374 by histology. RESULTS: Presence or absence of inflammation was concordantly diagnosed in 310 segments (77.3%). The highest concordance was found for the terminal ileum and, in patients with previous surgery, the anastomosis. Sensitivity and specificity for MRI versus endoscopy, MRI versus histology and endoscopy versus histology were 64.4%/81.1%, 62.1%/86.2% and 78.2%/80.3%, respectively. CONCLUSIONS: In a retrospective analysis of patients with Crohn's disease, hydro-MRI assessment of inflammation in anatomical segments of the colon reaches acceptable concordance rates with endoscopy and histology without prior preparation of the bowel. The data justify a prospective controlled trial. PMID- 17918004 TI - Dynamic colonization of Helicobacter pylori in human gastric mucosa. AB - OBJECTIVE: To investigate the dynamic behaviour of Helicobacter pylori in the colonization of the human gastric mucosa in patients previously treated for H. pylori infection. MATERIAL AND METHODS: Twenty-one dyspeptic patients were included in the study. Biopsies from each individual were taken and analysed for H. pylori detection using cultural, molecular and ultrastructural methods. RESULTS: Through culture, H. pylori was isolated in 7 out of 21 patients and the detection of the minimum inhibitory concentration (MIC) against drugs commonly used in H. pylori therapy revealed a susceptibility panel in which only one strain was multidrug resistant. By studying the expression of the H. pylori glmM constitutive gene, viable H. pylori cells were detected in 19 out of 21 analysed biopsies. In these positive cases, the expression of the Quorum-Sensing related gene, luxS, was always detected. The analysis of glmM and luxS sequences confirmed the H. pylori identity. Scanning electron microscopy (SEM) analysis of biopsies from patients harbouring culturable bacteria showed a prevalent "S shape" H. pylori morphotype co-existent with coccoid aggregated bacteria embedded in an abundant matrix; while samples from patients shown as H. pylori-positive only through the molecular method showed clustered coccoid bacteria arranged in a microbial biofilm. CONCLUSIONS: In the present work we describe a new scenario in H. pylori mucosa colonization suggesting, in infection recalcitrance, the planning of more efficacious protocols in order also to identify camouflaged and protected clustered bacteria, taking into account this serious microbial problem in medicine in the recommendation of therapeutic regimens. PMID- 17918008 TI - Is there an association between coeliac disease and inflammatory bowel diseases? A study of relative prevalence in comparison with population controls. AB - OBJECTIVE: The relationship between coeliac disease and inflammatory bowel disease (IBD) is controversial. The aim of this study was to determine the prevalence of coeliac disease in IBD and the prevalence of IBD in coeliac disease. MATERIAL AND METHODS: Patients were enrolled from specialist IBD and coeliac clinics. Antigliadins, endomysial, tissue transglutaminase antibody and total IgA levels were measured in IBD patients. Patients with positive antibodies were offered a duodenal biopsy. The notes on coeliac patients were reviewed for colonoscopic and biopsy findings. Controls were recruited from the local population. RESULTS: The study included 305 patients with coeliac disease, 354 with IBD and 601 healthy controls. The IBD group comprised 154 ulcerative colitis (UC) cases, 173 Crohn's disease, 18 indeterminate colitis and 9 cases of microscopic colitis. Forty-seven patients had positive antibodies and 3 had villous atrophy on biopsy. All three patients had positive anti-tissue transglutaminase antibodies but only two were endomysial antibody (EMA) positive. Ten coeliac patients had IBD (5 UC and 5 lymphocytic colitis). Five controls had coeliac disease and 2 had IBD (1 Crohn's disease and 1 UC). Stepwise multiple logistic regression showed only antibody positivity as being significant (p<0.0001). CONCLUSIONS: The prevalence of IBD in coeliac disease was increased 10-fold compared with that in controls (odds ratio 9.98, 95% CI 2.8-45.9, p=0.0006), while the prevalence of coeliac disease in IBD was comparable with that in controls (odds ratio 1.02, 95% CI, 0.24-4.29, p=1.0). PMID- 17918009 TI - Acute gastrointestinal bleeding from focal duodenal lymphangiectasia. PMID- 17918010 TI - Autoantibodies in patients with gut motility disorders and enteric neuropathy. AB - OBJECTIVE: Enteric neuropathy with mild inflammation (ganglionitis) has been described in several motility disorders including irritable bowel syndrome (IBS), enteric dysmotility (ED), slow-transit constipation (STC) and chronic intestinal pseudo-obstruction (CIPO). The purpose of this study was to test the hypothesis that autoantibodies directed against specific neural antigens including ion channels may be associated with this finding. MATERIAL AND METHODS: Comprehensive routine and immunohistochemical analyses of full-thickness jejunal laparoscopic biopsies were performed on patients fulfilling the international criteria for IBS, ED, STC and CIPO. Patients with ganglionitis had sera screened for specific antibodies to voltage-gated calcium channels (VGCCs) of P/Q- and N-type, voltage gated potassium channels (VGKCs), glutamic acid decarboxylase (GAD) and neuronal alpha3-AChR by validated immunoprecipitation assays. RESULTS: Thirty-three patients were included in the study. Two of them, both with IBS, were found to have positive antibody titres. One had anti-VGKC antibodies and one had anti alpha3-AChR antibodies. No antibodies were detected in GAD or VGCCs (case reports presented). CONCLUSIONS: A small proportion of patients with inflammatory enteric neuropathy have antibodies directed towards neuronal ion channels. The pathogenic role of such antibodies requires determination but may represent a possible aetiology of severe functional symptoms in these groups of patients. PMID- 17918011 TI - Prevalence and incidence of primary biliary cirrhosis are increasing in Finland. AB - OBJECTIVE: To examine the epidemiology of primary biliary cirrhosis (PBC) in Finland and to evaluate whether the possible increase in prevalence was attributable to the increasing incidence, better survival, or both. MATERIAL AND METHODS: The Hospital Discharge Register, pathology registers, and death certificates for the years 1988 99 were scrutinized, and the patients identified were followed-up for survival until 31 October 2004. The study area covered four university hospital districts: a total of 25 hospitals. The diagnosis of PBC was regarded as definite (or probable) if three (or two) of the following criteria were fulfilled: positive antimitochondrial antibodies, constantly elevated alkaline phosphatase, and compatible liver histology. RESULTS: In the total population of the study areas, the age-standardized prevalence of PBC increased during the study period from 103 (95% CI: 97-110) to 180 (172-189) per million inhabitants. Incidence increased from 12 (10-14) to 17 (15-20) per million inhabitants per year. The annual average increase in prevalence was 5.1% (4.2 5.9%, p <0.0001) and in incidence 3.5% (0.9%-6.0%, p =0.008). In gender-specific analyses among women, the prevalence of PBC increased from 161 (151-171) to 292 (277-207) per million during the study period and the incidence from 20 (16-24) to 27 (23-32) per million per year. The death rate was 4% per year and half the deaths were from liver-related causes. Survival after diagnosis during the study period lengthened. CONCLUSIONS: The prevalence of PBC increased in Finland during 1988-99, owing to both the increased incidence and the prolonged survival. PMID- 17918012 TI - Leucocyte apheresis in the treatment of paediatric ulcerative colitis. PMID- 17918013 TI - Elevated levels of high mobility group box chromosomal protein-1 (HMGB-1) in sera from patients with severe bacterial pneumonia coinfected with influenza virus. AB - Plasma levels of high mobility group box chromosomal protein-1 (HMGB-1), as well as of other inflammatory molecules such as interleukin-6 (IL-6), regulated on activation normal T-cell expressed and secreted (RANTES), and soluble intercellular adhesion molecule-1 (sICAM-1), were determined in patients with bacterial pneumonia coinfected with influenza virus. HMGB-1 levels were significantly elevated in these patients compared to patients undergoing mild bacterial pneumonia alone (p < 0.01). Among cases of coinfection, we found a significant correlation between the concentration of HMGB-1 and white blood cell counts (p < 0.05, r = 0.612). Levels of IL-6 were also higher in these patients than in patients with bacterial pneumonia alone (p < 0.05), despite similar levels of RANTES and sICAM-1 in the 2 groups. These data suggest that HMGB-1 is involved in the pathogenesis of severe bacterial pneumonia coinfected with influenza virus. PMID- 17918014 TI - Antimicrobial susceptibility of invasive group B streptococcal isolates from south-west Sweden 1988-2001. AB - The antibiotic susceptibility of 297 invasive isolates of group B streptococci (GBS) to a panel of 12 antibiotics was analysed using the E-test. The isolates (from 123 neonates and 174 adults) were collected from south-west Sweden during the 2 periods 1988-1997 and 1998-2001. The breakpoints of the Clinical and Laboratory Standards Institute were used. All isolates were sensitive to cefotaxime, meropenem, linezolid, vancomycin, moxifloxacin and quinupristin dalfopristin. Two strains displayed a slightly decreased susceptibility to penicillin G (MIC 0.25 microg/ml) also when tested by the broth dilution method. Two per cent were resistant to erythromycin and 1% to clindamycin. Strains with intermediate sensitivity to erythromycin and clindamycin increased over the 2 study periods. 68% were resistant to doxycycline, and the resistance rate for doxycycline increased over the 2 study periods. No strain was resistant to trimethoprim-sulfamethoxazole. Serotype V dominated among strains with intermediate susceptibility to erythromycin and clindamycin. There were no other relationships between serotypes and decreased sensitivity to any agent. There were no significant differences in susceptibility to any agent tested between strains isolated from neonates and adults. In conclusion, penicillins remain the drug of choice in the region but with the increasing rates of intermediate susceptibility to both erythromycin and clindamycin, antibiotic sensitivity analysis should be performed on the GBS isolates from penicillin-allergic patients. PMID- 17918015 TI - Febrile neutropenia in children with cancer: a retrospective Norwegian multicentre study of clinical and microbiological outcome. AB - Our objective was to describe clinical and laboratory characteristics, treatment and outcome among Norwegian children with cancer suffering from chemotherapy induced febrile neutropenia (FN). We retrospectively reviewed data on paediatric FN episodes in 7 Norwegian hospitals during a 2.5-y period. A total of 236 episodes of FN occurred in 95 children. Acute lymphoblastic leukaemia was the most common diagnosis (49 patients). Blood cultures yielded growth in 39 episodes (17%). Primary empirical antibiotic regimens could be assigned to 2 main groups: 1) benzylpenicillin or ampicillin and an aminoglycoside (58%) or 2) a regimen based on third-generation cephalosporins (42%). There were no statistically significant differences in outcome between the 2 regimens in terms of need to change initial antibiotic treatment, d of fever or maximum C-reactive protein values. One infection-related death (fungal septicaemia) occurred during the study period. We conclude that incidence of septicaemia and clinical outcome is similar to recent international trials on paediatric FN, but antibiotic treatment in Norway differs from international guidelines. However, patients in our study were successfully and safely treated, irrespective of the primary empirical antibiotic regimen. PMID- 17918016 TI - Contribution of molecular tools for the diagnosis of endogenous pneumococcal endophthalmitis in a patient with the acquired immunodeficiency syndrome. AB - Confirmation of infectious endophthalmitis is a major challenge. Traditional microbial tools are frequently unsuccessful mostly due to very small fluid samples and prior use of antibiotics. Here we report the first case of Streptococcus pneumoniae endophthalmitis diagnosed by polymerase chain reaction. Molecular biology should be considered as a new tool for the diagnosis of such sequestered sites as the eye. PMID- 17918017 TI - Comparing the pneumonia severity index with CURB-65 in patients admitted with community acquired pneumonia. AB - Pneumonia severity assessment systems such as the pneumonia severity index (PSI) and CURB-65 were designed to direct appropriate site of care based on 30-d mortality. Increasingly they are being used to guide empirical antibiotic therapy and also possibly to detect patients who will require admission to the intensive care unit (ICU). We retrospectively reviewed the records of all patients admitted to our institution with confirmed community acquired pneumonia (CAP) for the 12 months from January 2002. 408 episodes were studied with an overall 30-d mortality of 15.4% and ICU admission of 10.5%. PSI classes IV/V were significantly better than CURB-65 score > or = 3 for predicting patients who died within 30 d (94% vs 62%; p < 0.001), and those that needed ICU (86% vs 61%; p = 0.01). In addition, for the patients identified as 'low risk' by PSI (classes I/II), there was only 1 death and 1 admission to an ICU compared to 8 deaths and 7 ICU admissions with CURB-65 scores of 0-1. Although easier to use, CURB-65 is neither sensitive nor specific for predicting mortality in CAP patients. Neither rule was sufficiently accurate for predicting need for an ICU, even when patients with 'not for resuscitation' orders were excluded. PMID- 17918018 TI - The impact on community acquired pneumonia empirical therapy of diagnostic bronchoscopic techniques. AB - The aim of the present study was to examine the modification of initial empirical treatment based on the microbiological results of bronchoscopic techniques after comparing the diagnostic yield of protected specimen brush (PSB) and bronchoalveolar lavage (BAL) in the immunocompetent patient with community acquired pneumonia (CAP) with results obtained from conventional sputum cultures. 88 patients with presumptive diagnosis of CAP necessitating hospitalization were prospectively studied. Fibreoptic bronchoscopy with quantitative PSB and BAL cultures for common pathogens, mycobacteria and fungi was performed. Conventional sputum cultures were also obtained. PSB and BAL quantitative cultures added 26.1% and 36.4%, respectively, more microbiological documentation for CAP compared to conventional sputum cultures (p < 0.0001). Gram staining was indicative of the pathogen mostly in cases where Streptococcus pneumoniae was isolated, which was also the most frequently isolated pathogen (19.3%), followed by Haemophilus influenzae (9%). M. tuberculosis was isolated in 6.8% of patients. Modification of treatment ensued in 27.3% of patients because of the application of the cultures of sputum and invasive technique. PSB and BAL added significant information to the aetiological diagnosis of hospitalized immunocompetent patients with CAP. PMID- 17918019 TI - Neutrophil and monocyte receptor expression in uncomplicated and complicated influenza A infection with pneumonia. AB - Following influenza, the elderly and those with chronic heart/lung diseases are often affected by bacterial complications such as pneumonia. Whether neutrophil and monocyte functions are affected differently in patients with or without complications is less well known. Therefore, blood neutrophil and monocyte surface receptor expressions were measured in patients with influenza A, with or without complications, by means of flow cytometry. Neutrophil expressions of the adhesion molecules CD11b and CD66b were increased in influenza A, with the highest expression of CD11b in uncomplicated influenza. Monocyte expressions of CD11b and CD18 were also higher in influenza compared with bacterial infection and healthy controls. Neutrophil expressions of the phagocyte receptors CD64 and CD32 and the complement receptor CD35 were impaired in influenza with and without pneumonia compared with bacterial infection, whereas the expressions in monocytes were increased in all infected groups. The expression of the phagocyte receptor CD16 on neutrophils was impaired in all infected groups. Our results suggest increased recruitment of neutrophils and monocytes to infected areas by up regulation of adhesion molecules in influenza that may be involved in the inflammatory response during infection. In contrast, depression of phagocyte receptor expression on neutrophils in patients with influenza pneumonia may contribute to increased susceptibility to bacterial infections and impaired clearance of encapsulated bacteria such as pneumococci. PMID- 17918021 TI - Alcohol and drug consumption, depressive features, and family violence as associated with complaints to the Prosecutor's Office in Central Mexico. AB - The article is aimed at reporting the characteristics of the population detected at State Prosecutors' Offices including the two such offices that existed in the city selected for the study, one located in a general hospital for the inspection of violence-related cases (n = 156); and the second in the facility where all detainees are taken when arrested (n = 129), and where victims can file a complaint (n = 186). A household survey undertaken among the population 18 to 65 years of age (n = 887) was used as a group of reference. Both studies were undertaken in Pachuca City, the capital of Hidalgo, located 100 km from Mexico City during the second half of 1996. Face-to-face questionnaires were used to obtain sociodemographic data, drug use and drinking patterns, depressive symptomatology, and family violence. Discriminant and logistic regression analysis were undertaken. The age group from 18 to 24 displayed the highest number of legal complaints and arrests (OR = 1.773). The likelihood for appearing at a State Prosecutor's Office was higher for those living in an atmosphere of threats and injuries within the family (OR = 19) and for those that reported alcohol consumption on the day of the event (OR = 14). Extremely high rates of family violence were obtained in this sample, increasing the likelihood of arriving at the Prosecutor's Office either because arrested or for being a victim. Results confirm the relationship between alcohol use, depression, and violence, reinforcing the need to prevent alcohol abuse, especially among youth. PMID- 17918022 TI - Adolescent substance-use assessment: methodological issues in the use of the Adolescent Drug Abuse Diagnosis (ADAD). AB - During the past twenty years, various instruments have been developed for the assessment of substance use in adolescents, mainly in the United States. However, few of them have been adapted to, and validated in, French-speaking populations. Consequently, although increasing alcohol and drug use among teenagers has become a major concern, the various health and social programs developed in response to this specific problem have received little attention with regard to follow-up and outcome assessment. A standardized multidimensional assessment instrument adapted for adolescents is needed to assess the individual needs of adolescents and assign them to the most appropriate treatment setting, to provide a single measurement within and across health and social systems, and to conduct treatment outcome evaluations. Moreover, having an available instrument makes it possible to develop longitudinal and transcultural research studies. For this reason, a French version of the Adolescent Drug Abuse Diagnosis (ADAD) was developed and validated at the University Child and Adolescent Psychiatric Clinic in Lausanne, Switzerland. This article aims to discuss the methodological issues that we faced when using the ADAD instrument in a 4-year longitudinal study including adolescent substance users. Methodological aspects relating to the content and format of the instrument, the assessment administration and the statistical analyses are discussed. PMID- 17918023 TI - Behavioral strategies to improve treatment participation and retention by pregnant drug-dependent women. AB - The present study examined the utility of behavioral incentives for improving early treatment participation and retention in a sample of 91 pregnant opiate and/or cocaine dependent women enrolled in an urban, university-based drug user treatment program between 1996 and 1997. An escalating voucher incentive system was compared to standard care. The relationship between treatment participation and retention and maternal and infant outcomes were examined using logistic regression, chi-square analyses, and t-tests. Behavioral incentives did not decrease rates of very early dropout from residential treatment, although improved outpatient treatment participation and retention during the transition from residential care was noted. Behavioral strategies demonstrate utility as adjuncts to counseling for high-risk substance dependent patients. They appear ineffective, however, for improving early residential treatment participation and retention, suggesting other variables (e.g., psychiatric comorbidity) may be operating during the first 24-48 hours post treatment admission. PMID- 17918024 TI - Psychosocial factors in alcohol use-related problems of working youth. AB - This study investigated the psychosocial correlates of alcohol use related problems in a sample of 581 working adolescents (N = 4405), recruited from five vocational schools in Ankara in June 2004 with the CAGE questionnaire, The Beck Depression Inventory, the Beck Hopelessness Scale, the Spielberger State Anxiety Scale, and the Coopersmith Self-Esteem Inventory. Using a multivariate analysis, the anxiety and hopelessness scores, and the length of stay in Ankara were found to be related to alcohol-use problems of the working youth. The study's limitations were noted and future research was suggested. PMID- 17918025 TI - Assessment of families who have substance abuse issues: those who maltreat their infants and toddlers and those who do not. AB - Although an association exists among risk factors of age of child, substance use and child maltreatment, less is known about the differences between substance abusing families who maltreat their children and substance abusing families who do not maltreat their children. The relationship among substance use and maltreatment needs to be further explored to identify specific assessment techniques to discriminate between the two groups. The study findings, based upon case records analyzed in 2002, show there is a set of significant characteristics of families who have a history of substance use and who maltreat their infant and toddlers and those families who do not. PMID- 17918026 TI - Preference for sweet foods and higher body mass index in patients being treated in long-term methadone maintenance. AB - Opiate use has been associated with preference for sweets in both humans and animals. In 2002, the food preferences and eating habits of non-institutionalized patients in methadone treatment and controls were measured and compared. Questionnaires were administered to 14 patients and 14 controls with similar demographic characteristics (there were 19 women and 9 men ranging in age from 19 to 59 years). The patients reported higher consumption of sweets, higher eagerness to consume sweet foods, and a wish to consume quantities larger than that desired by controls. Patients had a significantly higher body mass than controls. The study's limitations are noted. These findings suggest a link between methadone and a desire for calorically dense foods. A study of eating behavior is warranted. PMID- 17918027 TI - Monitoring ecstasy content in France: results from the National Surveillance System 1999-2004. AB - The French National Identification System for Drugs and Other Substances (SINTES) is an original scheme gathering analytical information for synthetic drugs, both through police and customs' seizures in the entire country and collection of samples and questionnaires directly from the users by social field workers. Between July 1999 and June 2004, 9543 samples were included. Tablets (7004) were mainly containing MDMA (82%) and caffeine was the most frequent blended psychoactive substance. Mean MDMA dosage of tablets decreased from 1999 to 2003 and dosage for tablets bearing the same logo appeared to be highly variable. Notwithstanding the difficulties for data collection due to the illicit nature of these drugs, this surveillance and early warning system, which combines the cooperative efforts of law enforcement laboratories and social workers, provided relevant and timely information. It is accurate regarding the follow-up of trends in drugs' composition, and the identification of new or potentially dangerous substances, to the professionals, the public, and the European partners. PMID- 17918028 TI - A strategy for improving community effectiveness of HIV/AIDS intervention design: the Community Readiness Model in the Caribbean. AB - The Community Readiness Model (CR) offers a solution to the development of prevention strategies at the community level that makes use of community collaborations and community capacity. CR was administered in the U.S. Virgin Islands on St. Thomas, St. Croix, and St. John. All three islands scored lowest on dimensions of community climate and highest on knowledge of efforts to fight HIV/AIDS. The findings suggest that initial intervention efforts should improve the community climate of HIV/AIDS denial and stigma prior to introducing interventions that address HIV/AIDS risk behaviors. Community collaborators identify the interplay of social, structural, and political-economic factors for effective prevention. PMID- 17918029 TI - Pilot study: does the Five Shot Questionnaire give an indication of the severity of alcohol use-related problems? AB - This study explores whether the Five Shot Questionnaire reflects the severity of alcoholic inpatients' problems, such as assessed clinically by means of the EuropASI. Data were collected from 101 patients admitted to a psychiatric hospital in Belgium for alcohol use-related problems between September 2003 and December 2004. To investigate the research question, regression analyses--linear, ordinal, and binary logistic regression--have been performed. We conclude that the Five Shot Questionnaire gives an indication of the severity of alcohol use related problems. As such, it can be recommended in general practice and emergency departments for further evaluation and treatment planning. The article ends with a discussion of its limitations and some suggestions for future research. PMID- 17918030 TI - The damage done: a study of injection drug use, injection related abscesses and needle exchange regulation. AB - This study investigated the impact of a needle exchange policy change on community health. Data were collected from a needle exchange program in Eureka, California, for clients who participated in the program between the weeks (n = 112) of January 1, 2002, and February 28, 2004. Analysis was done using an autoregressive integrated moving average (ARIMA), indicating that greater utilization of the needle exchange program, in terms of needles exchanged and number of visits, is related to fewer numbers of abscesses treated. Additionally, self-report data collected from former intravenous drug users (n = 62) demonstrated that more needles exchanged were related to fewer occurrences of abscesses. The limitations of this research are discussed. PMID- 17918032 TI - PhySIC: a veto supertree method with desirable properties. AB - This paper focuses on veto supertree methods; i.e., methods that aim at producing a conservative synthesis of the relationships agreed upon by all source trees. We propose desirable properties that a supertree should satisfy in this framework, namely the non-contradiction property (PC) and the induction property (PI). The former requires that the supertree does not contain relationships that contradict one or a combination of the source topologies, whereas the latter requires that all topological information contained in the supertree is present in a source tree or collectively induced by several source trees. We provide simple examples to illustrate their relevance and that allow a comparison with previously advocated properties. We show that these properties can be checked in polynomial time for any given rooted supertree. Moreover, we introduce the PhySIC method (PHYlogenetic Signal with Induction and non-Contradiction). For k input trees spanning a set of n taxa, this method produces a supertree that satisfies the above-mentioned properties in O(kn(3) + n(4)) computing time. The polytomies of the produced supertree are also tagged by labels indicating areas of conflict as well as those with insufficient overlap. As a whole, PhySIC enables the user to quickly summarize consensual information of a set of trees and localize groups of taxa for which the data require consolidation. Lastly, we illustrate the behaviour of PhySIC on primate data sets of various sizes, and propose a supertree covering 95% of all primate extant genera. The PhySIC algorithm is available at http://atgc.lirmm.fr/cgi-bin/PhySIC. PMID- 17918031 TI - Amphetamine use and sexual risk among men who have sex with men: results from the National HIV Behavioral Surveillance study--New York City. AB - A 2004-2005 survey among 503 men who have sex with men who attend public venues in New York City was used to examine the relationship of amphetamine use with sexual risk behaviors. Among the men recruited, 51.1% were under 30 years of age, 27.4% were Latino, and 23.3% were African American. Most identified as either gay (78.9%) or bisexual (18.1%). A standardized questionnaire collected data on demographics, sexual risk behaviors, drug and alcohol use, history of HIV testing, and occurrences of sexually transmitted infections. Amphetamine use in the past year was reported by 13.8%. Of those, 71.0% used amphetamines with sex. Amphetamine use was associated with unprotected receptive anal intercourse with non-main partners. In event-specific analysis, amphetamine use was higher with unprotected encounters compared with protected encounters. This study confirms the association between amphetamine use and sexual risk furthers our understanding of risky circumstances and lays the groundwork for the design of interventions. PMID- 17918033 TI - The role of second trimester ultrasound in the diagnosis of placental hypoechoic lesions leading to poor pregnancy outcome. AB - OBJECTIVE: The purpose of the study was to quantify placental hypoechoic areas without blood flow by means of second trimester high-resolution ultrasound with power color Doppler and to evaluate its relationship to pregnancy outcome. METHODS: Patients referred for second trimester ultrasound from January 2001 to December 2003 were eligible for the study. Patients with placental parenchymal hypoechoic areas without blood flow constituted the study group (N = 65). The control group was comprised of 65 patients who had normal placental parenchyma. The groups were similar with regards to maternal age, gestational age, parity, race, and smoking. The mean of the two largest lesion diameters was used for severity classification. RESULTS: Fifty-two patients (80%) had aggregate lesions of 5 cm. Mean +/- standard deviation (SD) gestational age at delivery was 39.1 +/- 1.8 weeks for the control group, 37.9 +/ 2.8 weeks for those with lesions of 5 cm (p < 0.0001). Mean birth weight +/- SD was 3348 +/- 492 g for the controls, 3134 +/- 657 g for those with lesions of 5 cm (p = 0.0005). The incidence of intrauterine growth restriction was 9.6% in the group with lesions of 5 cm, compared to 3.1% in the control group (p = 0.0003; odds ratio (OR) = 5.7, p = 0.015). The incidence of preeclampsia in the control group was 0.0%, in the group with lesions of 5 cm was 15.4% (p < 0.0214; OR = 14.3, p = 0.014). CONCLUSIONS: Increased size of hypoechoic placental lesions is associated with increased risk for adverse perinatal outcome. Such lesions might be the result of intervillous space thrombosis, and ultrasound may be a useful tool in the identification of patients with pro-thrombotic abnormalities. The capability to identify patients with placental thrombosis will help us to identify those patients who will benefit most from anti-thrombotic treatment PMID- 17918037 TI - The effects of sequence difficulty and practice on proportional and nonproportional transfer. AB - Two experiments were designed to determine participants' ability to transfer a learned movement sequence to new spatial locations. A 16-element dynamic arm movement sequence was used in both experiments. The task required participants to move a horizontal lever to sequentially projected targets. Experiment 1 included two groups. One group practised a relatively easy 16-element movement sequence (easy long). The other group practised a more difficult 16-element movement sequence (difficult long). Approximately 24 hours after practice with their respective sequence both groups were administered a retention and two transfer tests. The only difference between the retention and transfer tests was the location of the targets. The short transfer target configuration was considered a proportional transfer because all the amplitudes between targets were reduced by the same proportion. The mixed transfer configuration was considered a nonproportional transfer because the targets did not have the same proportional distances between targets as the sequence they practised. The results indicated that participants could effectively transfer the difficult long sequence to the new target configurations regardless of whether the transfer required proportional and nonproportional spatial changes to the movement pattern. However, the easy long sequence was only effectively transferred in the proportional transfer condition. Experiment 2 assessed the effects of extended practice of the easy long sequence on proportional and nonproportional spatial transfer. The data indicated that participants could again effectively transfer the easy long sequence to proportional but not the nonproportional spatial transfer conditions regardless of the amount of practice (1 or 4 days). The results are discussed in terms of the mechanism by which response sequences become increasingly specific over extended practice in an attempt to optimize movement production and how this process interacts with the difficulty of the sequence. PMID- 17918038 TI - Bioactive phenols from the leaves of Baccaurea ramiflora. AB - Two new phenols, 6'- O-vanilloylisotachioside ( 1) and 6'- O-vanilloyltachioside ( 2), together with nine known compounds, were isolated from the leaves of Baccaurea ramiflora (Euphorbiaceae). The structures of the new compounds were elucidated mainly by analysis of physical and spectroscopic data. Compounds 1 - 10 were tested for antioxidant activities by using MTT and DPPH assays. Seven compounds, 1, 2, and 4 - 8, revealed potent antioxidant activities against H (2)O (2)-induced impairment in PC12 cells, and exhibited significant DPPH radical scavenging activities with IC (50) values of 86.9, 142.9, 15.2, 37.6, 35.9, 30.2, and 79.8 microM, respectively. PMID- 17918039 TI - Protective effect of St. John's wort (Hypericum perforatum) extract on 72-hour sleep deprivation-induced anxiety-like behavior and oxidative damage in mice. AB - Sleep disruption or poor quality of sleep is a common problem associated with depression. Antidepressant drugs have been reported to improve the quality of sleep and behavior. The present study was undertaken to explore the therapeutic potential of Hypericum perforatum (St. John's wort) on behavioral alterations and oxidative damage induced by sleep deprivation in mice. Male laca mice (n = 6 - 10 in each group) were sleep deprived for 72 hours using the grid suspended over water method. Standardized Hypericum perforatum extract and imipramine were administered for five days, starting two days before sleep deprivation. Alterations in body weight, motor activity, anxiety like behavior (mirror chamber, plus maze, zero maze) and oxidative stress parameters (reduced glutathione, catalase, lipid peroxidation and nitrite levels) were observed after drug treatment in sleep-deprived animals. 72-hour sleep deprivation significantly altered body weight, locomotor activity and produced anxiety-like behavior and oxidative damage (depleted reduced glutathione, catalase activity and increased lipid peroxidation and nitrite activity) as compared to the naive (placed on sawdust) animals (P < 0.05). Treatment with either St. John's wort (200 and 400 mg/kg, P. O.) or with imipramine (10 mg/kg, I. P.) significantly improved body weight, locomotor activity, antianxiety and antioxidant effect as compared to the control group (sleep deprived) (P < 0.05). Co-administration of John's wort (200 mg/kg, P. O.) with imipramine (10 mg/kg, I. P.) further improved body weight, locomotor activity, antianxiety effect as well as reduced oxidative damage in sleep-deprived animal as compared to their effect per se (P < 0.05). The present study suggests that there is therapeutic potential of St. John's wort in the management of sleep deprivation-induced anxiety-like behavior and oxidative damage. PMID- 17918040 TI - Andrographolide inhibits human hepatoma-derived Hep3B cell growth through the activation of c-Jun N-terminal kinase. AB - Andrographolide (Andro) is a potentially anti-inflammatory diterpenoid lactone isolated from the traditional herbal medicine ANDROGRAPHIS PANICULATA, which has been effectively used for the treatment of infection, inflammation, cold, fever and diarrhea in China for centuries. In the current study, we found that Andro significantly decreased the number of surviving hepatoma-derived Hep3B cells in the MTT assay and induced cell apoptosis. Further study showed that Andro induced activation of mitogen-activated protein kinases (MAPKs) including p38 kinase, c Jun N-terminal kinase (JNK) and extracellular signal-related kinases (ERK1/2), but had no significant effect on caspase-3, Bcl-xL and Bcl-2, which are apoptosis related proteins. Moreover, inhibition of JNK activation partially rescued the toxic effect of Andro on Hep3B cells. Therefore, our results indicate that the JNK signaling pathway plays an important role in the toxic effect of Andro on Hep3B cells. PMID- 17918042 TI - Duplex sonography of hemodialysis access. PMID- 17918041 TI - Gastroprotective effect of mangiferin, a xanthonoid from Mangifera indica, against gastric injury induced by ethanol and indomethacin in rodents. AB - In search of novel gastroprotective agents, mangiferin, a naturally occurring glucosylxanthone from Mangifera indica L. (Anacardiaceae), was evaluated in mice on gastric injury induced by ethanol and indomethacin. The effects of mangiferin on gastric mucosal damage were assessed by determination of changes in mean gastric lesion area or ulcer score in mice and on gastric secretory volume and total acidity in 4-h pylorus-ligated rats. Mangiferin (3, 10 and 30 mg/kg, P. O.) significantly attenuated the gastric damage induced by ethanol by 30, 35, and 63 %, and of indomethacin by 22, 23 and 57 %, respectively. N-Acetylcysteine (750 mg/kg, I. P.) and lansoprazole (30 mg/kg, P. O.) used as positive controls in these ulcerogenic models resulted in 50 % and 76 % suppression of gastric injury, respectively. In 4-h pylorus-ligated rats, intraduodenally applied mangiferin (30 mg/kg) caused significant diminutions in gastric secretory volume and total acidity. In addition, like N-acetylcysteine, a donor of sulfhydryls, mangiferin effectively prevented the ethanol-associated depletion of gastric mucosal non protein sulfhydryl content in mice, suggesting an antioxidant action. These findings provide evidence that mangiferin affords gastroprotection against gastric injury induced by ethanol and indomethacin most possibly through the antisecretory and antioxidant mechanisms of action. PMID- 17918043 TI - How to write and publish a successful scientific article. PMID- 17918044 TI - Effect of knee position on ultrasound Doppler findings in patients with patellar tendon hyperaemia (jumper's knee). AB - PURPOSE: The purpose of this study is to see if changes in patient positioning result in significant changes in Doppler findings. MATERIALS AND METHODS: 30 consecutive patients with jumper's knee in the proximal portion of the patellar tendon formed the study group. The patellar tendon was scanned in the longitudinal plane with colour and spectral Doppler with the knee fully extended as well as, flexed at 15 degrees and 20 degrees . All subjects were randomised to either extension or 20 degrees flexion as the initial position. The amount of colour Doppler activity inside the tendon was expressed with the colour fraction (colour pixels/total pixels) in the proximal portion of the tendon. With spectral Doppler, the resistive index was measured ((peak systolic velocity--end-diastolic velocity)/ peak systolic velocity). RESULTS: The mean (+/- SE) colour fraction in the fully extended position and 20 degrees flexion was 36.7 +/- 3.4% and 13.3 +/- 3.4%, respectively (p<0.0001). The mean resistive index was 0.607 +/- 0.036 with full extension and 0.914 +/- 0.036 with 20 degrees flexion (p<0.0001). On an individual patient basis, a decrease in colour fraction was observed in all 30 patients (100%), whereas an increase in resistive index was observed in 29 patients (97%). Maximum perfusion was seen in all positions when flexion was the initial position. CONCLUSION: We advocate that for the examination of the patellar tendon by Doppler sonography, the patients should have fully extended and relaxed knees. Otherwise, the colour Doppler findings will underestimate the flow, and the spectral Doppler will overestimate peripheral vascular resistance. PMID- 17918045 TI - Quality management of nuchal translucency measurement in residents. AB - PURPOSE: The measurement of the nuchal translucency (NT) in the 1st trimester is a sensitive, reliable method to assess the risk of specific fetal chromosomal and other defects. Training, however, is an issue not only among experienced sonographers, but especially for ObGyn residents, since all NT measurements in a true screening setting should fulfil the quality standards. The aim of this study was therefore the evaluation of the learning curve of residents and determination of the number of measurements necessary to achieve acceptable results. MATERIALS AND METHODS: Between 30th June, 1997 and 8th August, 2003, we included 4450 subsequent pregnant women between 11+0 and 13+6 weeks of gestation referred for an NT scan and prenatal counselling (low and high risk patients) in the study. For analysis of the learning curve in residents, all NT scans performed either by the experienced sonographers only or by residents with less than 70 scans at the end of their training were excluded. As the main quality criterion, the percentage of cases above the median was used. To test for normal distribution of NT scans, the Kolmogorov-Smirnov test was used. RESULTS: Each of 19 residents fulfilling these criteria performed 131 NT scans (73-242) on average. 13 of 19 residents ultimately met the quality criteria, but the majority of residents achieved good quality only after 100 scans, whereas 6 of 19 never did. Only after at least 50 NT scans, 50% of measurements were above the median, whereas before these 50 scans, NT was usually underestimated. CONCLUSION: It became obvious that regular supervision and quality control is mandatory to provide exact NT measurements by residents. Based on our results, a minimum of 100 NT scans is recommended before diagnostic application, which is a higher requirement than implemented in widely accepted quality assurance programs. PMID- 17918046 TI - [Dynamic transrectal ultrasound (dTRUS): a new method to diagnose anastomotic insufficiency after radical retropubic prostatectomy]. AB - PURPOSE: To prove extravasation after radical retropubic prostatectomy (RRP) nowadays a cystoradiogram is essential. In the present study the diagnostic value of dynamic transrectal ultrasound and cystoradiogram to find an extravasation was compared. MATERIALS AND METHODS: For detection of an extravasation Cystoradiogram and dTRUS were performed in 250 patients who had undergone RRP. TRUS was performed dynamically, that means that the urinary bladder was filled up with common salt solution backwards by using the transurethral catheter. Anastomosis was inspected by transrectal ultrasound. Detectable extravasation was measured and documented. To verify the results a cystoradiogram was carried out afterwards. In cases of detectable extravasation the results of measurement were compared to the results of dTRUS. RESULTS: The mean age was 65 years. An extravasation could be detected in 46% within the first 7 days and in 18% after 14 days. At day 21 an extravasation could not be seen in any patient. Seven days postoperative the mean volume of extravasation was 11 ml (3-50 ml) and after 14 days 9 ml (3-25 ml) for dTRUS. For cystoradiogram 12 ml (3-45 ml) and 9 ml (4-23 ml), respectively. The average time until catheter removal was 8 days (5-35 days). 60% of the patients were continent immediately after removing the catheter, 40% were incontinent for not more than 3 months after removal of the catheter (35% ICS I and 5% ICS II). A prolonged urinary incontinence and serious postoperative complications were not observed. CONCLUSION: Dynamic transrectal ultrasound is a reliable and reasonable method to identify extravasation after RRP. Furthermore radiation exposure (on average 60 cGy/cm(2)/cystoradiogram) can be avoided by replacing the cystoradiogram with dTRUS slips with the dTRUS. PMID- 17918047 TI - Current status of intraoperative real-time vibrography in neurosurgery. AB - PURPOSE: Intracranial lesions are often characterized by different elasticities. The aim of the present study was to test the application of vibrography during brain tumor surgery. MATERIALS AND METHODS: The real-time vibrography system consisted of a conventional ultrasound system (Siemens Sonoline Omnia) with a custom-designed RF interface and a 6.5-MHz endocavity curved array (Siemens 6.5EC10). The RF data were digitized using a 50-MHz, 12-bit PCI analog/digital (A/D) converter for real-time or offline processing. Static compression was replaced by low-frequency axial vibration of the probe. A special applicator equipped with a stepping motor moved the ultrasonic probe and produced a low frequency mechanical vibration of 5-10 Hz with a vibration amplitude of 0.3 mm and slight preliminary compression (total<1 mm). The maximum application time was 60 sec. A pneumatic holding device (Unitrac, Aesculap, Tuttlingen, Germany) was used. RESULTS: Brain tissue is normally color coded between red and orange. In this study 41 out of 45 tumors could be detected via vibrography. Two tumors could not be detected with this imaging technique: a glioblastoma at a depth of 2 cm and a metastasis at a depth of 3 cm. Two additional tumors were not recognized because of technical problems. In 4 cases tumors with strain values identical to those in brain tissue (coded red or orange) but easily identified by a peripheral zone of high strain (yellow) were found. Tumors with strain values higher than those measured in brain tissue coded yellow and were softer than brain during surgical intervention. Higher strain was found in 23 tumors. Tumors with strain values lower than those in cerebral tissue were found to be harder during surgery and coded brown or black. Lower strain was found in 10 cases. Four tumors were inhomogeneous and could not be assigned to one of the above groups. Mortality was 0%, morbidity 2.3%. One patient displayed transient paresis of the lower extremity due to microsurgical difficulties during the approach. In one patient minimal bleeding of the cortical surface occurred in a frontobasal tumor; however, no postoperative deficits were noted. CONCLUSION: Vibrography is a new low-risk technique for intraoperative imaging. In low-grade astrocytomas and oligodendrogliomas, this additional technique can be used to control resection. In other cortical and subcortical tumors (e. g. metastases), it can provide an impression of the intratumoral elasticities. PMID- 17918049 TI - [Sonographic diagnosis of caudal regression syndrome]. AB - Caudal regression sequence (CRS) is a rare developmental defect of the lower spinal segments and the neural tube. Motor and sensory neurological deficits of the lower extremities as well as a reduced control of bowel and bladder functions are the main symptoms. Etiology and pathogenesis are widely unknown. This article discusses a newborn male with postnatal anomalies of the lower extremities. Sonographically, the spinal cord ended in the lumbar region. NMR confirmed the suspected sonographic finding of CRS. Sonographic examination of the abdominal vessel system depicted a common origin and junction of the hepatic artery, splenic artery and superior mesenteric artery from one common truncus. This points to a possible relict of a persisting vitelline artery. As previously described in sirenomelia, the findings in the present case indicate a possible vascular etiology of CRS. PMID- 17918050 TI - [Epidemiology of cutaneous leishmaniasis in Diyarbakir (2002-2006)]. AB - It is estimated that more than 1.5 million new cutaneous leishmaniasis (CL) cases occur throughout the world every year and the disease is endemic in southern and southeastern Turkey. The aim of this study was to evaluate the epidemiological characteristics of the CL cases reported in Diyarbakir. A total of 1990 cases were reported during the 5 year period. CL was most frequent in women (25.6 in 100.000) and in the age groups 5-9 and 10-14 (32.0 and 40.3 in 100.000 respectively). CL was most often reported in the Dicle (52.9%) and Hani (37.7%) districts; most were reported in the spring (47.2%) and fewer in the autumn (11.6%). It was observed that reporting increased after diagnosis and the quality of health services improved. Morbidity in Diyarbakir was higher than in the rest of Turkey. The higher prevalence found in children and in women was compatible with other studies. A difference from the other studies was that while morbidity decreased in the 15-44 age-group, there was an increase in the 45 or more age group. CL is endemic in the province of Diyarbakir. The fight against CL must be supported by special programs. Health education of the public and self protection measures should be encouraged along with a greater accessibility of diagnosis and treatment. PMID- 17918051 TI - Detection of Babesia (Theileria) equi (Laveran, 1901) in horses in the Kars province of Turkey. AB - This study was carried out in order to detect antibodies to Babesia (Theileria) equi in the local breed of horses in the province of Kars, Turkey. A total of 108 serum samples from apparently healthy horses in eight villages were examined for B. equi antibodies by an indirect immunofluorescent antibody test (IFAT). Of the 108 samples tested, 27 (25%) were found to be seropositive. The horses sampled in Aydinalan village had the highest prevalence (50.0%) of Babesia equi infection while the lowest prevalence was found among horses from Bayraktar village (12.5%). Statistically significant differences in seroprevalence were observed between these two villages (P < 0.05). This study is the first report on the status of B. equi infection in Kars. PMID- 17918052 TI - [PCR-RFLP analysis of the Tams1 gene of Theileria annulata]. AB - Tams1 is a merozoite surface antigen of Theileria annulata. Genetical variations of Tams1 make studies of vaccine and diagnostic tests like ELISA difficult. In this study, Tams1 genes of 89 T. annulata isolates obtained from natural infected cattle in Elazig and Bingol regions were tested with PCR-RFLP. Six different restriction profiles (a, b, c, d, e, f) were detected. The number of restriction profiles of 89 samples was found to be as follows: 78(a), 2(b), 2(c), 5(d), 1(e), and 1(f). PMID- 17918053 TI - [The seroprevalence of Toxoplasma gondii in women from Sanliurfa, a province with a high raw meatball consumption]. AB - Toxoplasma gondii is an obligate intracellular protozoan that can infect all kind of birds and all mammals including humans and is common throughout the world. The prevalence varies according to social and cultural habits, pet cats in homes and geographic factors. Domestic cats are considered to be an important source of Toxoplasma gondii infection. From January to June 2006, the prevalence of toxoplasmosis was retrospectively monitored from blood samples that had been sent to our laboratory in order to determine the levels of IgM and IgG. All the subjects were women and 2,586 blood samples were investigated with the chemiluminescence immunoassay method. The rates of Toxoplasma gondii IgM antibodies were found to be 3.0% (78/2,586) and that of Toxoplasma gondii IgG antibodies, 69.5% (1.798/2,586). The total rate of positivity of Toxoplasma gondii antibodies was 69.6% (1,801/2,586) and the negativity, 30.4% (785/2,586). The highest positive rates have been reported in the southeastern region of Turkey and the 69.6% detected in our study seems to be the highest rate. Raw meatball consumption is common in our region and raw meat has a high risk of Toxoplasma gondii infection by direct ingestion of tissue cysts. As a result we consider that the high frequency of Toxoplasma gondii seropositivity in this region is due to raw meatball consumption. PMID- 17918054 TI - A novel procedure for total nucleic acid extraction from small numbers of Eimeria species oocysts. AB - A series of experiments were performed in an attempt to extract genomic DNA from a small number of Eimerian oocysts. Sonication, ammonia, ethanol and lysozyme were all found to be unsuitable for the digestion of Eimeria oocysts. The chemicals and enzyme given were not capable of either disruption or digestion of oocysts for nucleic acid extraction. They had the capability of penetrating the oocyst wall but could not break-up the oocyst wall. It is impossible to obtain nucleic acid from Eimeria oocysts if the wall is not broken-up. In this study oocyst disruption was achieved using a simple but highly effective treatment regime involving sodium hypochlorite treatment, osmotic shock and proteinase K digestion. Following the disruption of the oocyst walls, a commercially available nucleic acid purification kit (Wizard DNA Purification Kit, Promega) can be used to prepare high quality nucleic acid. PMID- 17918055 TI - Pathogenicity of Blastocystis hominis, a clinical reevaluation. AB - Blastocystis (B.) hominis was considered to be a member of normal intestinal flora in the past, but in recent years it has been accepted as a very controversial pathogenic protozoan. In this study, 52 individuals whose stool examination revealed B. hominis were evaluated for clinical symptoms. Metronidazole was administered for 2 weeks to the patients infected with B. hominis. After 2 weeks of treatment they were called for a follow-up stool examination. No other bacteriological and parasitological agents were found during stool examination of these patients. The frequency rate of intestinal symptoms was 88.4% in the B. hominis cases. Abdominal pain was the most frequent symptom (76.9%). Diarrhea and distention followed at a rate of 50.0% and 32.6%. Intestinal symptoms may be seen frequently together with the presence of B. hominis and this protozoan may be regarded as an intestinal pathogen, especially when other agents are eliminated. PMID- 17918057 TI - The first report in Turkey of in vivo cultivation in Rattus norvegicus of Echinococcus multilocularis human strain. AB - The adult form of the small cestode Echinococcus multilocularis is found in carnivorous animals, especially in the fox. This cestode, which is found in the northern hemisphere in the world, is the cause of a generally fatal disease in humans, known as alveolar echinococcosis (AE). The metacestodes of Echinococcus multilocularis can be experimentally developed in the Meriones unguiculatus species of rodents, and it is possible to use these metacestodes for numerous purposes, primarily for supplying the antigens required for serological diagnosis of the disease. It is with this aim that in this study for the first time in Turkey, Echinococcus multilocularis metacestodes were developed using different kind of rodent, Rattus norvegicus, and an indigenous strain was isolated using the surgical material of a patient diagnosed with alveolar echinococcosis. PMID- 17918056 TI - [The evaluation of the techniques used for diagnosis of Entamoeba histolytica in stool specimens]. AB - In this study, stool samples of 9378 patients from different clinics, who presented at the laboratory of the department of parasitology of the Dokuz Eylul University, Faculty of Medicine with several gastrointestinal complaints from January 2004 to May 2006, were examined. All stool samples were examined with the saline-Lugol method and, in suspicious cases, by trichrome staining, cultivation in Robinson's medium and/or antigen detection in stool with the Entamoeba CELISA Path kit. Forty-one cases (0.44%), in which Entamoeba histolytica/Entamoeba dispar cysts and/or trophozoites were detected by at least one method, were found to be positive. Out of these 41 cases, four methods were used in 24 cases, three methods in 14 cases, whereas only saline-Lugol and trichrome staining methods were used in 3 cases. Even though all 41 positive cases had been examined with the saline-Lugol method, only 25 cases were found to be positive with this method for E. histolytica/E. dispar cysts and/or trophozoites. The remaining 16 cases were diagnosed by the other three methods. Today it is necessary to distinguish E. histolytica from E. dispar because the patient does not need to be treated if E. dispar is identified whereas if E. histolytica is identified the patient needs urgent treatment. That's why it is necessary to get reliable results using diagnostic methods together and, when needed, by ELISA specific for E. histolytica. PMID- 17918058 TI - [Examination of helminth contaminated wastewaters used for agricultural purposes in Afyonkarahisar]. AB - This research was performed in order to examine helminth contaminated wastewaters used in agriculture in Afyonkarahisar. Ten rural areas were selected that used water for agricultural purposes from a creek contaminated with human and animal wastes. Between September 2004 and August 2005, these areas were visited once every two weeks, and a total of 240 water samples were collected. Out of untreated water samples, 32 samples (26.70%) had helminth eggs, whereas there were neither helminths nor developmental stages of helminths (p < 0.001) in treated water samples. Cestode eggs were seen in 10.00% of untreated water samples and nematode eggs were seen in 16.70%. Out of the helminth eggs, 13 (40.60%) were hookworm eggs; 6 (18.75%), Taenia spp.; 5 (5.60%), Ascaris lumbricoides; 3 (9.40%), Hymenolepis diminuta; 3 (9.40%), H. nana; and 2 (6.25%), Toxocara spp. In conclusion; the creek was found to be contaminated by helminths originating from human and animal wastes in Afyonkarahisar. This type of water presents a risk for human and animal health and contaminated waters, therefore, should be avoided in irrigation of vegetables and fruit consumed raw. PMID- 17918059 TI - [A retrospective analysis of the results of a seven-year parasitological examination of stools from Malatya State Hospital]. AB - The aim of this study was to determine the prevalence of intestinal parasites and to evaluate changes in the annual rates by retrospective screening of the data from our hospital's parasitology laboratory between the period of 2000 and 2006. Parasitological examinations were done with native-Lugol, cellophane-tape methods and trichrome staining in ambiguous cases, in a total of 67539 samples. The prevalence of the intestinal parasitosis during the years 2000 and 2006 years were found to be 7.2% and 3.8%, respectively. The most prevalent parasites were Entamoeba spp. (except for E. histolytica) (53%), Giardia intestinalis (25.9%) and Enterobius vermicularis (8.3%). Entamoeba histolytica/E. dispar (5.3%), Ascaris lumbricoides (3%), Taenia saginata (2.2%) and Hymenolepis nana (2%) were the other parasites detected in this study. Even though a decrease in the annual prevalence of parasitosis was apparent, it was noticed that the parasitic diseases are still a significant health problem in our country. PMID- 17918060 TI - [Screening of intestinal parasites of children in special day nurseries in the city of Rize]. AB - In this study, the carriage of intestinal parasites was investigated in a total of 73 children (35 girls, 38 boys) in the 1-6 age-group in two special day nurseries in the city of Rize. Stool samples and cellophane tape preparations were obtained from children three times a month. Parasite cysts or eggs were found in total of 15.0% of the stool samples or cellophane tape preparations from children. It has been determined that 8.5% of the girls and 21.0% of the boys were parasite porters, and that all of these were asymptomatic carriers. Giardia intestinalis, Entamoeba coli + Iodamoeba butschlii, Taenia spp. and Enterobius vermicularis were detected at rates of 11.0%, 1.3%, 1.3% and 1.3%, respectively. PMID- 17918061 TI - [The occurrence of gastrointestinal cestode and nematode infections in stray dogs in Afyonkarahisar and Eskisehir provinces]. AB - This study was carried out to determine the prevalence of cestode and nematode infections in stray dogs in Afyonkarahisar and Eskisehir provinces. A total of 287 dogs (n=150 from Afyonkarahisar and n=137 from Eskisehir) of different ages and sexes living in animal shelters were used in this study. After the coprological examination, dogs were found to be infected with various gastrointestinal cestode and nematode species. It was found that rates of infection were 46% (69 out of 150) in Afyonkarahisar and 33.6% (46 out of 137) in Eskisehir provinces. Species responsible for the infection were the same in both provinces. In the Afyonkarahisar province 59.4% hookworms, 47.8% Toxascaris leonina, 36.2% Toxocara canis, 2.9% Dipylidium caninum and 2.9% Taenia spp. were detected. In the Eskisehir province 60.9% Toxascaris leonina , 47.8% Toxocara canis, 23.9% Taenia spp., 6.5% hookworms and 4.3% Dipylidium caninum were detected. It was concluded that in Afyonkarahisar and Eskisehir provinces, stray dogs might be infected with helminths which put humans in danger and necessary measures are needed. PMID- 17918062 TI - [Treatment of Linognathus vituli and Bovicola bovis infestations of calves with cypermethrin]. AB - This study has been conducted to determine the efficacy level of cypermethrin on the 14 calves that were naturally infested by Linognathus vituli and Bovicola bovis. A 10% solution of cypermethrin was diluted to 0.1% and applied to infested animals twice. All L. vituli were found to be dead after the fourth day of the first application and the second day of the second application. After the first day of application, all of the B. bovis were found dead at the end of each treatment. This study confirmed the protective efficacy of cypermethrin against L. vituli and B. bovis showing that infestation is prevented for a period of about 24-35 days following administration of cypermethrin at this dose. PMID- 17918063 TI - [A case of Diplotriaena monticola (Yamaguty, 1935) in the white-spectacled bulbul (Pycnonotus xanthopygos)]. AB - In Hatay, 13 nematodes (8 [female symbol] and 5 [male symbol]) were observed in the thoracic cavity of a white-spectacled bulbul (Pycnonotus xanthopygos). The parasites were cleared with lactophenol and then investigated under the light microscopy and important morphological parts of parasites were measured. The parasites were identified as Diplotriaena monticola Yamaguty, 1935 (Fam.: Filariidae) under the subfamily of Diplotriaeninae. It is important because this case is the first report of this infection in a white-spectacled bulbul in Turkey. PMID- 17918064 TI - [Investigation of house dust mite incidence related to social factors]. AB - House dust mites are found everywhere there are temperatures of 25-27 degrees C and humidity of 70-80 percent. The amount of acari varies according to social conditions. The aim of this study was to determine the relationship between social factors and the observable rate of acari. A total of 303 dust samples were examined and according to results of questionnaires, increase of acari populations was observed under the following conditions: crowded homes, wooden houses, using second hand furniture, poor ventilation, insufficient cleaning, no vacuum cleaner, lack of sunlight indoors, high humidity, and presence of pets in the garden. Even though the acari density was sometimes found to be high, relationships with factors were not statistically significant. Advice to allergic patients about precautions concerning the factors may be useful. PMID- 17918065 TI - [Helminth infections of camels]. AB - Camels, which are indispensable under Asia and Africa's bad climatic and geographic conditions, are bred in Turkey for tourist purposes including wrestling. Even though they have been used for thousands of years, comprehensive studies addressing the parasitic diseases of these animals have been performed only during recent years. In this review, helminth infections seen in the camel and their treatment have been discussed. PMID- 17918066 TI - [Helminth fauna of Neurergus strauchii (Steindachner, 1888) (spotted salamander) collected in Malatya and Bitlis]. AB - Forty-one (23 females, 18 males) Neurergus strauchii (spotted salamanders) collected in Malatya and Bitlis, were examined for helminths in 2001-2002. In this study, 1 helminth species, Cosmocerca commutata (Nematoda) was found. This study is the first helminthological research on spotted salamanders in the world. Also, this is the first time the parasite of frogs has been reported in Neurergus strauchii. PMID- 17918067 TI - Helminth infections in common carp, Cyprinus carpio L., 1758 (Cyprinidae) from Kovada Lake (Turkey). AB - The aim of this study which was carried out from March 2003-February 2004 was to determine the parasites of carp (Cyprinus carpio L., 1758) inhabiting the Kovada Lake. During the study, a total of 63 common carps were caught in different regions of Kovada Lake each month and investigated parasitologically. In carps, the ectoparasite, Dactylogyrus minutus of Monogenea, was found and endoparasites; Bothriocephalus acheilognathi and Caryophyllaeus laticeps of Cestoda were found. The most common parasitic species was Dactylogyrus minutus. PMID- 17918068 TI - Report on the monogenean Cyclocotyla bellones and three cymothoids parasitizing two fish species from the Aegean Sea coasts of Turkey. AB - In this paper, three separate instances of an association between an ectoparasite, Cyclocotyla bellones (Otto, 1821) (Monogenea: Diclidophoridae) and ectoparasitic isopods have been reported for the first time from the Aegean Sea coasts of Turkey. The isopods were all of the family Cymothoidae: Ceratothoa oestroides, C. parallela, Emetha audouini. PMID- 17918069 TI - First report of Pennella balaenopterae Koren and Danielssen, 1877 (Copepoda: Pennelidae) from Turkey. AB - A parasitic copepod, Pennella balaenopterae Koren and Danielssen, 1877 (Copepoda: Pennelidae), has been reported from the fin whale, Balaenoptera physalus (Cetacaea: Mysticeti), which was found dead on the Avluk beach (Yumurtalik seaport, Iskenderun Bay, Northeastern Mediterranean coast of Turkey) in 6 May, 2002. This is the first report of Pennella balaenopterae in the Turkish seas. PMID- 17918070 TI - Fatal brain abscess due to community-associated methicillin-resistant Staphylococcus aureus strain USA300. AB - We report a fatal case of brain abscess caused by infection due to a community associated methicillin-resistant Staphylococcus aureus strain (USA300) in a 37 year-old incarcerated woman with a history of furunculosis and injection drug use. Community-onset pyogenic brain abscess should be added to the growing list of life-threatening invasive infections caused by epidemic community-acquired methicillin-resistant S. aureus. PMID- 17918071 TI - Human T lymphotropic virus type 1-associated myelopathy/tropical spastic paraparesis in an HIV-positive patient coinfected with human T lymphotropic virus type 2 following initiation of antiretroviral therapy. AB - We describe a patient coinfected with human immunodeficiency virus (HIV) and human T lymphotropic virus type 2 in Spain who developed paraparesis resembling human T lymphotropic virus type 1-associated myelopathy/tropical spastic paraparesis shortly after initiation of highly active antiretroviral therapy, hypothetically as the result of an immune reconstitution inflammatory syndrome. PMID- 17918072 TI - Culture-negative prosthetic joint infection. AB - BACKGROUND: Culture-negative (CN) prosthetic joint infection (PJI) has not been well studied. We performed a retrospective cohort study to define the demographic characteristics and determine the outcome of patients with CN PJI. METHODS: All cases of CN total hip arthroplasty and total knee arthroplasty infections (using a strict case definition) treated at our institution from January 1990 through December 1999 were analyzed. Kaplan-Meier survival methods were used to determine the cumulative probability of success. RESULTS: Of 897 episodes of PJI during the study period, 60 (7%) occurred in patients for whom this was the initial episode of CN PJI. The median age of the cohort was 69 years (range, 36-87 years). Patients had received a prior course of antimicrobial therapy in 32 (53%) of 60 episodes. Of the 60 episodes, 34 (57%), 12 (20%), and 8 (13%) were treated with 2 stage exchange, debridement and retention, and permanent resection arthroplasty, respectively. The median duration of parenteral antimicrobial therapy was 28 days (range, 0-88 days). Forty-nine (82%) of 60 episodes were treated with a cephalosporin. The 5-year estimate of survival free of treatment failure was 94% (95% confidence interval, 85%-100%) for patients treated with 2-stage exchange and 71% (95% confidence interval, 44%-100%) for patients treated with debridement and retention. CONCLUSIONS: CN PJI occurs infrequently at our institution. Prior use of antimicrobial therapy is common among patients with CN PJI. CN PJI treated at our institution is associated with a rate of favorable outcome that is comparable to that associated with PJI due to known bacterial pathogens. PMID- 17918074 TI - National trends in Staphylococcus aureus infection rates: impact on economic burden and mortality over a 6-year period (1998-2003). AB - BACKGROUND: We evaluated historical trends in the Staphylococcus aureus infection rate, economic burden, and mortality in US hospitals from 1998 through 2003. METHODS: The Nationwide Inpatient Sample was used to assess trends over time of S. aureus infection during 1998-2003. Historical trends were determined for 5 strata of hospital stays, including all inpatient stays, surgical procedure stays, invasive cardiovascular surgical stays, invasive orthopedic surgical stays, and invasive neurosurgical stays. RESULTS: During the 6-year study period from 1998 through 2003, the rate of S. aureus infection increased significantly for all inpatient stays (from 0.74% to 1.0%; annual percentage change (APC), 7.1%; P=.004), surgical stays (from 0.90% to 1.3%; APC, 7.9%; P=.001), and invasive orthopedic surgical stays (from 1.2% to 1.8%; APC, 9.3%; P<.001). For invasive neurosurgical stays, the rate of S. aureus infection did not change from 1998 to 2000 but increased at an annual rate of 11.0% from 2000 to 2003 (from 1.4% to 1.8%; P=.034). The total economic burden of S. aureus infection for hospitals also increased significantly for all stay types, with the annual percentage increase ranging from 9.2% to 17.9% (P<.05 for all). In 2003, the total economic burden of S. aureus infection was estimated to be $14.5 billion for all inpatient stays and $12.3 billion for surgical patient stays. However, there were significant decreases in the risk of S. aureus-related in-hospital mortality from 1998 to 2003 for all inpatient stays (from 7.1% to 5.6%; APC, 4.6%; P=.001) and for surgical stays (from 7.1% to 5.5%; APC, -4.6%; P=.002). CONCLUSIONS: The inpatient S. aureus infection rate and economic burden of S. aureus infections for US hospitals increased substantially from 1998 to 2003, whereas the in-hospital mortality rate decreased. PMID- 17918073 TI - Genotype prevalence and risk factors for severe clinical adenovirus infection, United States 2004-2006. AB - BACKGROUND: Recently, epidemiological and clinical data have revealed important changes with regard to clinical adenovirus infection, including alterations in antigenic presentation, geographical distribution, and virulence of the virus. METHODS: In an effort to better understand the epidemiology of clinical adenovirus infection in the United States, we adopted a new molecular adenovirus typing technique to study clinical adenovirus isolates collected from 22 medical facilities over a 25-month period during 2004-2006. A hexon gene sequence typing method was used to characterize 2237 clinical adenovirus-positive specimens, comparing their sequences with those of the 51 currently recognized prototype human adenovirus strains. In a blinded comparison, this method performed well and was much faster than the classic serologic typing method. RESULTS: Among civilians, the most prevalent adenovirus types were types 3 (prevalence, 34.6%), 2 (24.3%), 1 (17.7%), and 5 (5.3%). Among military trainees, the most prevalent types were types 4 (prevalence, 92.8%), 3 (2.6%), and 21 (2.4%). CONCLUSIONS: For both populations, we observed a statistically significant increasing trend of adenovirus type 21 detection over time. Among adenovirus isolates recovered from specimens from civilians, 50% were associated with hospitalization, 19.6% with a chronic disease condition, 11% with a bone marrow or solid organ transplantation, 7.4% with intensive care unit stay, and 4.2% with a cancer diagnosis. Multivariable risk factor modeling for adenovirus disease severity found that age <7 years (odds ratio [OR], 3.2; 95% confidence interval [CI], 1.4-7.4), chronic disease (OR, 3.6; 95% CI, 2.6-5.1), recent transplantation (OR, 2.7; 95% CI, 1.3 5.2), and adenovirus type 5 (OR, 2.7; 95% CI, 1.5-4.7) or type 21 infection (OR, 7.6; 95% CI, 2.6-22.3) increased the risk of severe disease. PMID- 17918075 TI - Fluoroquinolone use and risk factors for Clostridium difficile-associated disease within a Veterans Administration health care system. AB - BACKGROUND: Prompted by the changing profile of Clostridium difficile infection and the impact of formulary policies in hospitals, we performed this study when an increase in the incidence of C. difficile-associated disease was noted at our health care center (Veterans Administration Puget Sound Health Care System, Seattle, Washington). METHODS: A retrospective, matched case-control study of patients presenting to the Veterans Administration Puget Sound Health Care System, Seattle, Washington during 2004 was performed. Conditional logistic analysis determined risk factors for case patients, defined as individuals with diarrhea and test results (i.e., culture or toxin assay results) positive for C. difficile, and control subjects, defined as individuals with diarrhea and test results negative for C. difficile. RESULTS: C. difficile-associated disease incidence was 29.2 cases per 10,000 inpatient-days. The increase in the incidence of C. difficile-associated diarrhea that paralleled increased gatifloxacin use was not attributable to use of the antimicrobial but was a reflection of seasonal variation in the rate of C. difficile-associated disease. Multivariate analysis controlling for the time at which the assay was performed, the age of the patient, ward, and source of acquisition (community-acquired vs. nosocomial disease) found 6 significant risk factors for C. difficile-associated diarrhea: receipt of clindamycin (adjusted odds ratio [aOR], 29.9; 95% confidence interval [CI], 3.58-249.4), receipt of penicillin (aOR, 4.1; 95% CI, 1.2-13.9), having a lower intestinal condition (aOR, 2.8; 95% CI, 1.3-6.1), total number of antibiotics received (aOR, 1.4; 95% CI, 1.1-1.7), number of prior hospital admissions (aOR, 1.3; 95% CI, 1.1-1.6), and number of comorbid conditions (aOR, 1.3; 95% CI, 1.1-1.5). CONCLUSIONS: The increase in the number of cases of C. difficile-associated disease was not attributable to a formulary change of fluoroquinolones; instead, the incidence was within expected seasonal variations for C. difficile-associated disease. Recognition of community-acquired cases and the use of culture may help to identify additional cases of C. difficile associated disease. Early diagnosis and treatment of C. difficile cases may shorten the duration of hospital stays and reduce the number of outbreaks and readmissions, mortality, and other consequences of C. difficile infection. PMID- 17918076 TI - Detection of toxigenic Clostridium difficile in stool samples by real-time polymerase chain reaction for the diagnosis of C. difficile-associated diarrhea. AB - BACKGROUND: Clostridium difficile-associated diarrhea (CDAD) is the major cause of health care-associated infectious diarrhea. Current laboratory testing lacks a single assay that is sensitive, specific, and rapid. The purpose of this work was to design and validate a sensitive and specific real-time polymerase chain reaction (PCR) diagnostic test for CDAD. METHODS: This observational validation study of a new real-time PCR assay occurred from July 2004 through April 2006 and involved the testing of 1368 stool samples. As the final validation portion of the investigation, 350 inpatients were prospectively interviewed for clinical findings for 365 episodes of diarrheal illness. Test results and clinical criteria were used to assess the performance of 4 assays. RESULTS: Using clinical criteria requiring at least 3 loose stools in 1 day as part of the reference standard for a positive test result supporting CDAD, the sensitivity, specificity, and positive and negative predictive values were 73.3%, 97.6%, 73.3%, and 97.6%, respectively, for enzyme immunoassay; 93.3%, 97.4%, 75.7%, and 99.4%, respectively, for real-time PCR; 76.7%, 97.1%, 69.7%, and 97.9%, respectively, for cell culture cytotoxin assay; and 100.0%, 95.9%, 68.2%, and 100.0%, respectively, for anaerobic culture (for toxigenic C. difficile strains). The real-time PCR and anaerobic culture assays were significantly more sensitive than the enzyme immunoassay (P<.01 to P<.05). CONCLUSIONS: With an assay turnaround time of <4 h, real-time PCR is a more sensitive and equally rapid test, compared with enzyme immunoassay, and is a feasible laboratory option to replace enzyme immunoassay for toxigenic C. difficile detection in clinical practice, as well as for use during the development of new therapeutic agents. PMID- 17918077 TI - Fungal infections in recipients of hematopoietic stem cell transplants: results of the SEIFEM B-2004 study--Sorveglianza Epidemiologica Infezioni Fungine Nelle Emopatie Maligne. AB - BACKGROUND: The purpose of our study was to evaluate the incidence and outcome of invasive fungal infection (IFI) among patients who underwent autologous or allogeneic hematopoietic stem cell transplantation (HSCT) at 11 Italian transplantation centers. METHODS: This cohort-retrospective study, conducted during 1999-2003, involved HSCT patients admitted to 11 tertiary care centers or university hospitals in Italy, who developed IFIs (proven or probable). RESULTS: Among 3228 patients who underwent HSCT (1249 allogeneic HSCT recipients and 1979 autologous HSCT recipients), IFI occurred in 121 patients (overall incidence, 3.7%). Ninety-one episodes (2.8% of all patients) were due to molds, and 30 (0.9%) were due to yeasts. Ninety-eight episodes (7.8%) occurred among the 1249 allogeneic HSCT recipients, and 23 (1.2%) occurred among the 1979 autologous HSCT recipients. The most frequent etiological agents were Aspergillus species (86 episodes) and Candida species (30 episodes). The overall mortality rate was 5.7% among allogeneic HSCT recipients and 0.4% among autologous HSCT recipients, whereas the attributable mortality rate registered in our population was 65.3% (72.4% for allogeneic HSCT recipients and 34.7% for autologous HSCT recipients). Etiology influenced the patients' outcomes: the attributable mortality rate for aspergillosis was 72.1% (77.2% and 14.3% for allogeneic and autologous HSCT recipients, respectively), and the rate for Candida IFI was 50% (57.1% and 43.8% for allogeneic and autologous HSCT recipients, respectively). CONCLUSIONS: IFI represents a common complication for allogeneic HSCT recipients. Aspergillus species is the most frequently detected agent in these patients, and aspergillosis is characterized by a high mortality rate. Conversely, autologous HSCT recipients rarely develop aspergillosis, and the attributable mortality rate is markedly lower. Candidemia was observed less often than aspergillosis among both allogeneic and autologous HSCT recipients; furthermore, there was no difference in either the incidence of or the attributable mortality rate for candidemia among recipients of the 2 transplant types. PMID- 17918078 TI - Complex clonal and plasmid epidemiology in the first outbreak of Enterobacteriaceae infection involving VIM-1 metallo-beta-lactamase in Spain: toward endemicity? AB - BACKGROUND: We report the emergence and spread of metallo-beta-lactamases (MBLs) among enterobacterial isolates at Ramon y Cajal University Hospital (Madrid, Spain). METHODS AND RESULTS: During the period from March 2005 through September 2006, 25 patients (52% of whom were in the intensive care unit) were infected and/or colonized with single or different MBL-producing Enterobacteriaceae isolates (Klebsiella pneumoniae, 14 patients; Enterobacter cloacae, 12 patients; Escherichia coli, 1 patient; and/or Klebsiella oxytoca, 1 patient). Clonal analysis (XbaI pulsed-field gel electrophoresis) revealed that all K. pneumoniae isolates belonged to the same clone, but 6 patterns were found among the E. cloacae isolates. Carbapenems were affected to different degrees (minimum inhibitory concentration, < or = 1 to > 8 microg/mL), as were aminoglycosides and ciprofloxacin. The bla(VIM-1) MBL gene was present in all isolates; in addition, the bla(SHV-12) extended-spectrum beta-lactamase gene was detected in K. pneumoniae and E. coli isolates. The bla(VIM-1) gene was detected within a 4.0-kb class 1 integron (bla(VIM-1)-aacA4-dfrII-aadA1-catB2) in K. pneumoniae and E. coli and in a 2.5-kb class 1 integron (bla(VIM-1)-aacA4-aadA1) in E. cloacae and K. oxytoca isolates. The bla(VIM-1) gene was transferable (filter-mating) in 14 of 14 K. pneumoniae isolates, 4 of 11 E. cloacae isolates, and 1 of 1 E. coli isolate. A 60-kb plasmid belonging to the IncI1 group was detected in the epidemic VIM-1-K. pneumoniae clone. Plasmids of 300- or 435-kb belonging to IncH12 group were found among E. cloacae isolates. CONCLUSIONS: K. pneumoniae-MBL monoclonal epidemics coexisted with E. cloacae-MBL multiclonal epidemics in our hospital. The spread of the bla(VIM-1) gene among Enterobacteriaceae was driven by clonal spread associated with intergeneric plasmid transfer with different class I integron platforms. Such complex epidemiology might anticipate endemicity and should be considered for the design of containment epidemiology strategies. PMID- 17918079 TI - A step closer to extreme drug resistance (XDR) in gram-negative bacilli. PMID- 17918080 TI - Patient knowledge and attitudes about antiviral medication and vaccination for influenza in an internal medicine clinic. AB - BACKGROUND: Despite the introduction of Centers for Disease Control and Prevention guidelines for their use, antiviral medications for influenza remain underutilized. Our objective in this study was to describe beliefs, attitudes, and knowledge regarding antiviral medication and vaccination for influenza among patients in an internal medicine clinic. METHODS: We conducted a cross-sectional survey of adult patients in an internal medicine clinic from April through June 2006. RESULTS: Two-hundred eighty patients completed the survey. Fifty-five percent received influenza vaccination for the most recent influenza season. Overall antiviral knowledge was poor. Of 8 antiviral knowledge questions, the mean percentage of correct answers was 40%; 1 (<1%) of the patients answered all questions correctly, and 47 (18%) answered all questions incorrectly. Only 37 (13%) of the patients reported calling their physician within 48 h after the onset of influenza-like symptoms. Patients with conditions associated with a high risk of complications from influenza were no more likely than other patients to be more knowledgeable about antiviral medication, nor were they more likely to report calling their physician within 48 h after symptom onset or to report receipt of influenza vaccination for the previous influenza season. Only 90 (37%) of the respondents were willing to pay >$20 for antiviral medication, although 205 (84%) were willing to pay something. CONCLUSIONS: Patients are ill-informed about antiviral medication and its benefits, and medication costs may present a barrier to treatment. Physicians should discuss antiviral medication with patients who are at high risk for complications from influenza before the influenza season, and education programs for physicians and patients should be developed. PMID- 17918081 TI - Improving care for acute respiratory infections: better systems, not better microbiology. PMID- 17918082 TI - Immune reconstitution syndrome and exacerbation of infections after pregnancy. AB - Pregnancy is a state of subtle immunosuppression characterized by physiologic suppression of proinflammatory host responses that are meant to promote embryonic implantation. Rapid reversal of these changes and a rebound of inflammatory responses during the postpartum period can result in quiescent or latent infection manifesting as symptomatic disease. Infections due to several microbial pathogens and noninfectious diseases with an autoimmune basis have been shown to worsen or begin during the postpartum period. Awareness that symptoms resulting from immune reconstitution can occur in any host with a rapidly changing immunologic repertoire, including women in the postpartum phase, is a critical first step in fully understanding this phenomenon. Future studies to discern the precise pathophysiologic basis of immune reconstitution, to identify pregnant women at risk, and to determine markers that may be diagnostically helpful have significant implications for optimizing treatment of these patients. PMID- 17918083 TI - Nationwide increase in the number of hospitalizations for illicit injection drug use-related infective endocarditis. AB - Infective endocarditis is a potentially fatal consequence of illicit injection drug use. We estimate that the number of hospitalization for injection drug use related infective endocarditis increased by 38%-66% in the United States between 2000-2001 and 2002-2003, a period during which the number of at-risk persons (i.e., injection drug users) remained stable. Increasing methamphetamine use and/or drug injection frequency may have increased the incidence of infective endocarditis among active injection drug users. PMID- 17918084 TI - The presence of dupA in Helicobacter pylori is not significantly associated with duodenal ulceration in Belgium, South Africa, China, or North America. AB - A previous study suggested that Helicobacter pylori strains possessing dupA are positively associated with duodenal ulceration and negatively associated with gastric adenocarcinoma. We determined the prevalence of dupA in H. pylori strains recovered from 4 independent populations and found a significant association with gastric cancer but not with duodenal ulceration. PMID- 17918085 TI - A 6-year-old boy with facial swelling and monocular blindness. PMID- 17918086 TI - Coinfection with HIV and tropical infectious diseases. I. Protozoal pathogens. AB - The brunt of the human immunodeficiency virus (HIV) pandemic has been borne disproportionately by resource-poor regions of the world, where tropical infectious diseases continue to hold greatest sway. As a result, our understanding of the epidemiological, biological, and clinical interactions between HIV and tropical pathogens has lagged, compared with our understanding of the interactions between HIV and pathogens that are common in the industrialized world. Because of the current rapid expansion of HIV care in the tropics, with increasing resources being made available, an overview of the available data is timely. Tropical protozoa are discussed here; other tropical pathogens are discussed in a related mini-review in this issue of Clinical Infectious Diseases. PMID- 17918087 TI - Coinfection with HIV and tropical infectious diseases. II. Helminthic, fungal, bacterial, and viral pathogens. AB - The morbidity, mortality, and social disruption caused by the human immunodeficiency virus (HIV) pandemic continue to weigh disproportionately on resource-poor regions of the tropics. As a result, the potential for significant epidemiological, biological, and clinical interactions between HIV and other tropical pathogens is great. An overview of the available data on tropical helminths, fungi, bacteria, and viruses is provided here; interactions between HIV and tropical protozoa are covered in a related mini-review in this issue of Clinical Infectious Diseases. Special attention is given to evidence relevant to the hypothesis that helminth coinfection plays a particularly important role in accelerating the pace of HIV pathogenesis in the tropics. PMID- 17918088 TI - Evolution of hepatitis B serological markers in HIV-infected patients receiving highly active antiretroviral therapy. AB - BACKGROUND: Evolution of serological markers of hepatitis B virus (HBV) carriage or infection has rarely been investigated among human immunodeficiency virus (HIV)-infected patients receiving highly active antiretroviral therapy (HAART). METHODS: During the period 1997-2002, a total of 633 HIV-infected patients were tested for HBV serological markers at baseline, including hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs ), antibody to hepatitis B core antigen (anti-HBc), hepatitis C virus (HCV) antibody (anti-HCV) antibody, HCV RNA level, and HBV DNA level, all of which were retested at least 1 year apart. Medical records were reviewed to identify clinical characteristics associated with evolution of these serological markers. RESULTS: After a median duration of follow-up for 4.96 years, 161 patients (25.4%) had changes in HBV serological markers. Of 119 patients (18.8%) who tested positive for HBsAg at baseline, 6 (5.0%) developed anti-HBs, and 9 (7.6%) developed isolated anti-HBc. Of 270 patients (42.7%) who tested positive for anti-HBs, 18 (6.7%) lost anti-HBs. Of 179 patients (28.3%) in whom isolated anti-HBc had been detected, 73 (40.8%) developed anti-HBs, 18 (10.1%) lost all HBV markers, and 7 (3.9%) developed HBsAg. Of 65 patients (10.2%) who tested negative for all HBV markers, 13 (20%) developed anti-HBs, 13 (20%) developed isolated anti-HBc, and 4 (6.2%) developed HBsAg, indicating a high risk of HBV exposure. Patients in whom anti-HBc was detected at baseline were more likely to have acquired immunodeficiency syndrome (P=.008). Multivariate analysis revealed that an increase in the CD4 cell count after the commencement of HAART was significantly associated with persistence or subsequent development of anti-HBs in patients with anti-HBs or anti-HBc at baseline, respectively. CONCLUSIONS: Periodic measurements of HBV serological markers in HIV-infected patients are recommended, because new HBV infections and changes of HBV serological markers are not uncommon in patients with improved immunity after commencement of HAART. PMID- 17918091 TI - Case-control studies to assess vaccine effectiveness? Yes, but not this way. PMID- 17918089 TI - Successful efavirenz dose reduction in HIV type 1-infected individuals with cytochrome P450 2B6 *6 and *26. AB - BACKGROUND: Efavirenz (EFV) is metabolized primarily by cytochrome P450 2B6 (CYP2B6), and high plasma concentrations of the drug are associated with a G-->T polymorphism at position 516 (516G-->T) of CYP2B6 and frequent central nervous system (CNS)-related side effects. Here, we tested the feasibility of genotype based dose reduction of EFV. METHODS: CYP2B6 genotypes were determined in 456 human immunodeficiency virus type 1 (HIV-1)-infected patients who were receiving EFV treatment or were scheduled to receive EFV-containing treatment. EFV dose was reduced in CYP2B6 516G-->T carriers who had high plasma EFV concentrations while receiving the standard dosage (600 mg). EFV-naive homozygous CYP2B6 516G-->T carriers were treated with low-dose EFV. In both groups, the dose was further reduced when plasma EFV concentration remained high. RESULTS: CYP2B6 516G-->T was identified in the *6 allele (found in 17.9% of our subjects) and a novel allele, *26 (found in 1.3% of our patients). All EFV-treated CYP2B6 *6/*6 and *6/*26 carriers had extremely high plasma EFV concentrations (>6000 ng/mL) while receiving the standard dosage. EFV dose was reduced to 400 mg for 11 patients and to 200 mg for 7 patients with persistently suppressed HIV-1 loads. EFV-containing treatment was initiated at 400 mg in 4 CYP2B6 *6/*6 carriers and one *6/*26 carrier. Two of them still had a high plasma EFV concentration while receiving that dose, and the dose was further reduced to 200 mg, with successful HIV-1 suppression. CNS-related symptoms improved with dose reduction in 10 of the 14 patients, although some had not been aware of the symptoms at initial dosage. CONCLUSIONS: Genotype-based EFV dose reduction is feasible in CYP2B6 *6/*6 and *6/*26 carriers, which can reduce EFV-associated CNS symptoms. PMID- 17918092 TI - Pitfalls in case-control studies of vaccine effectiveness. PMID- 17918094 TI - Polymorphisms at position 245 of HIV reverse transcriptase do not accurately predict the presence of human leukocyte antigen B*5701. PMID- 17918096 TI - The long-term outcomes of an antibiotic control program with and without education. PMID- 17918097 TI - Living with tuberculosis: the myths and the stigma from the Indian perspective. PMID- 17918099 TI - Postoperative central nervous system infection after neurosurgical procedures: the bride is too beautiful. PMID- 17918100 TI - Acute pyelonephritis: management steps that remain unresolved. PMID- 17918102 TI - [Prophylaxis of chronic daily headache with a simplified regimen of subcutaneous administration of botulinum toxin type A]. AB - INTRODUCTION: Primary headaches are one of the most prevalent conditions in neurology. Botulinum toxin type A (TBA) has been used for years as a prophylactic measure for chronic daily headache (CDH) with varying infiltration regimens and outcomes. AIM: To demonstrate the speed, convenience and safety of a subcutaneous regimen of TBA as a prophylactic for CDH, and also to determine how effective it is as a preventive therapy. PATIENTS AND METHODS: A total of 843 patients were infiltrated subcutaneously with 100 U of TBA (Botox) using a 'fixed dose-fixed site' technique in six pericranial points in periods of at least three months. We evaluated the reduction in the mean frequency of crises and days with headaches and the intake of analgesics, as well as the percentage of patients with a reduction of over 50% in the mean frequency of headache crises. RESULTS: Subcutaneous infiltration of TBA brought about a reduction of 78.91+/-6.81% in the frequency of monthly crises (p<0.001) and a reduction of 62.38+/-12.8% in the number of days per month with headache after three months' infiltration. Reports from patients indicated that 92.8% of them had reduced the number of days with headache per month by at least 50%. Mean frequency and total intake of analgesics decreased from 21.48+/-2.47 to 1.96+/-1.27 (p<0.001) and from 126.01+/-24.7 to 6.65+/-2.27 doses, respectively. There were few side effects, none of which were severe. CONCLUSION: Simplified subcutaneous infiltration of TBA is very effective, convenient, fast and safe as a means of preventing CDH. PMID- 17918103 TI - [Functional recovery in hemifacial transplants in rats]. AB - INTRODUCTION: There are a number of different options open to the surgeon for the reconstruction of the face and scalp, but when tissue loss is very extensive, good aesthetic and functional recovery is not possible. Not only must the damaged tissues be replaced, but motor and sensorial functioning also has to be restored. AIM: To evaluate the functional recovery of hemifacial allografts in rats. MATERIALS AND METHODS: Twenty-one hemifacial flaps were transplanted from Long Evans rats to Wistar-Lewis rats, under immunosuppression monotherapy with tacrolimus. Prior to the operation, anatomical and allograft viability studies were conducted. Two groups of transplanted rats were formed: with or without nerve repair. In the nerve repair group, end-to-end suture was employed to repair the infraorbital branch of the trigeminal nerve and the buccolabial, upper mandibular marginal and zygomatico-orbital branches of the facial nerve. Sensory recovery was evaluated by filming traction of the whiskers, whereas motor recovery was assessed by blind tests using electromyography studies of the mystacial muscles and electroneurography of the facial nerve. At eight weeks, the animals were sacrificed and biopsy samples were taken from the mystacial region. RESULTS: The facial flap was successfully lifted in 10 cases. In the nerve repair group both clinical and electrophysiological recovery were observed at six weeks, whereas biopsy samples taken in the eighth week showed recovery of the nerve fascicles. CONCLUSIONS: The hemifacial flap can be transplanted. By repairing the nerves of the allograft, it is possible to achieve its functional recovery, as can be confirmed clinically, electrophysiologically and histopathologically. To date, this is the first evidence of functional recovery following a hemifacial transplant in rats. PMID- 17918104 TI - [ADHD Rating Scale-IV in a sample of Spanish schoolchildren: normative data and internal consistency for teachers and parents]. AB - INTRODUCTION: The ADHD Rating Scale-IV (ADHD RS-IV) is one of the most use scales for attention deficit hyperactivity disorder (ADHD), because has cut-off point regarding age, gender and setting, but the normalization data is based on American school samples. AIM: Evaluation of cut-off point of ADHD-RS-IV for parents and teachers in a Spanish sample. SUBJECTS AND METHODS: The study used the score of ADHD-RS-IV from a prevalence study of ADHD in school children of 6 12 years. Using an intrasubject design between the three evaluators (parents and teachers) and the results of each subscale (IN, H/I, and TOT) according to gender and age factors. Then, we analyzed the reliability and internal consistency for each subscale and evaluator. RESULTS: There are no significant differences between the father and the mother; but there are between teachers, and father and/or mother scores. In relation to gender factor, boys' score is higher on inattention and hyperactivity-impulsivity than girls'. Our results show a reversal tendency in comparison with the American samples, in our case parents' scores were significantly higher than teachers'. CONCLUSION: The use and normalization of the ADHD RS-IV will need to account not only for age, gender and setting but also for socio-cultural aspects. PMID- 17918105 TI - [Comorbidity in the migraine: depression, anxiety, stress and insomnia]. AB - INTRODUCTION: Migraine is frequently associated to other pathologies or factors. AIM: To analyze the migraine's profile in a neurologic consultation and to determinate its comorbidity. PATIENTS AND METHODS: 155 patients (36 men and 119 women) were studied in a neurologic consultation of the University Hospital of Salamanca during the year 2005. Migraine was diagnosed according the International Headache Society criteria, and associated disorders were measured using the Goldberg's Anxiety and Depression Scale, the Perceived Stress Questionnaire and the Oviedo Sleep Questionnaire. RESULTS: 99 patients suffered from migraine without aura and 56 with aura; mean age was 36 years; age at onset 20.64 years. 43 migraneurs had depression, anxiety 34, insomnia 58, stress 79, none of these pathologies 62 (40%). Depression and dream upheavals were close to equal between the migraine subtypes (although depression was something more common in migraine without aura), while anxiety and stress were more frequent in migraine with aura (RR: 1.42; 95% CI: 0.92-2.2; and RR: 1.38; 95% CI: 0.9-2.12, respectively). Depression and insomnia were more likely among women with nonsignificant differences (RR: 1.3; 95% CI: 0.95-1.34; and RR: 1.13; 95% CI: 0.95-1.34, respectively), whereas anxiety and stress were similar in both genders. Another associations were found between: migraine's family-history and migraine with aura (OR: 1.31; 95% CI: 0.63-2.71); some dietary factors and migraine without aura (OR: 2.27; 95% CI: 0.57-9.07). CONCLUSIONS: 60% of our migraneurs had at least one of the following disorders: depression (27.74%), anxiety (21.94%), stress (50.97%) or insomnia (37.42%); can trigger or worsen migraine attacks. PMID- 17918106 TI - [Atypical presentation of intramedullary sarcoidosis: report of two cases]. AB - INTRODUCTION: Neurological involvement in sarcoidosis occurs in 5 to 15% of cases, but medullary involvement is very uncommon. CASE REPORT: We report two cases of sarcoidosis with medullary involvement as an initial presentation. Case 1: a 41 year-old man presented with a four-month history of lumbar pain, gait and sphincter disturbances; examination revealed a spastic paraparesis, hyperreflexia and a sensory level by D4-D6. Serum angiotensin converting enzyme (SACE) was normal. Magnetic resonance image (MRI) showed diffuse spinal cord enlargement with hyperintensity signal between D4-D5 and D10-11, with contrast enhancement. Spinal biopsy was consistent with sarcoidosis. Case 2: a 36 year-old man presented with a two-months history of gait difficulties and two episodes of blurred vision on his left eye that improved with topical therapy. Examination revealed lupus pernio in the ears and a spastic hyperreflexic paraparesis. Laboratory detected increased SACE. MRI showed focal spinal cord areas of hyperintensity at cervico-dorsal levels with contrast enhancement. Ear biopsy was consistent with sarcoidosis. Clinical courses under corticosteroids were favourable in both cases. CONCLUSION: Intramedullary lesions are infrequently the first manifestation of sarcoidosis especially when there is no systemic involvement (case 1). It is also rare in association with lupus pernio and ocular involvement (case 2). In the later (with systemic manifestations) SACE was raised. Biopsy is very useful for early diagnosis. PMID- 17918108 TI - [The genetics of child temperament]. AB - INTRODUCTION: In spite of the high initial expectancy in preliminary results concerning the genetics of personality, these studies have not provided satisfactory results. The failure could be related to the lack of biological validity of personality concept and the important influence of environmental factors on personality. A possible way to solve this problem is to look at the temperament of preschool children. It is expected that variability in infants' behaviour can be better defined and with less environmental influence. DEVELOPMENT: Firstly, twin and adoption studies of child temperament in comparison with the studies of personality in adults are reviewed. Secondly, the molecular association studies carried out concerning child temperament are analyzed. The serotonin transporter gene (5-HTT), D4 receptor gene (DRD4) and mono amino oxidade-A gene (MAOA) have been considered candidates to explain variability in child temperament because these genes have been related with specific personality dimensions and mental diseases. Finally, the methodological problems and the future direction of research in this field are considered. CONCLUSIONS: Heritability shows higher values in infant temperament than in adult personality. Different gene polymorphisms on 5-HTT, DRD4 and MAOA could explain some individual variability in children's behavior, although replication studies are needed to confirm the role of these genes. Longitudinal studies in large samples that include gene and environmental interactions are one of the best ways to improve our knowledge about the genetics of child temperament. PMID- 17918107 TI - [Memory coding and retention: brain-derived neurotrophic factor (BDNF) in synaptic plasticity]. AB - INTRODUCTION: It has been suggested that the long-term modifications that take place in synaptic transmission constitute the foundation of the processes by which information stored. The neurotrophin called brain-derived neurotrophic factor (BDNF) has recently emerged as a powerful molecular mediator in central synaptic plasticity. DEVELOPMENT: In this work we review the studies that have represented a significant step forward in explaining the role played by BDNF in long-term synaptic plasticity. The effects of BDNF on synaptic plasticity can be of a permissive nature, whereby it establishes the conditions under which plastic changes can take place, or it may be instructive. In this latter case it exerts direct effects that bring about changes in the communication and morphology of the synapses. The actions carried out by BDNF include its capacity to contribute to the stabilisation and maturation of already-existing synapses, as well as to generate new synaptic contacts. One important finding that highlights the participation of this neurotrophin in synaptic plasticity is the observation that adding BDNF gives rise to drastic long-term increases in synaptic transmission, similar to the long-term potentiation in the hippocampus and neocortex of mammals. CONCLUSIONS: Because BDNF modulates both the electrical properties and the structural organisation of the synapse, this neurotrophin has been considered to be an important marker during learning and memory processes. PMID- 17918109 TI - [A proposed new target for deep brain stimulation in obsessive-compulsive disorder]. AB - INTRODUCTION: The obsessive-compulsive disorder (OCD) has an incidence in general population of 1.5-3%. If we consider as a positive respond a diminution of the 25 35% in the symptoms of OCD according to the Y-BOCS, and we add the cognitive behavioral therapy to the pharmacological treatment, only a 40-60% of treated patients would have significant improvement and a 10% of patients with OCD, would be refractory to all type of medical treatment. DEVELOPMENT: Current neurosurgical techniques for resistant cases of OCD interrupt the connections between the frontal lobes and subcortical structures (cingulotomy, capsulotomy). These techniques are ablative and irreversible. It shows the importance of finding a less aggressive technique with better clinical results. Deep brain stimulation (DBS) is an alternative to traditional neurosurgery based in neuromodulation methods. It's considered that the physiopathology of the OCD consists of a dysfunction of the direct and indirect vias that control the extrapiramidal limbic circuit. On the other hand, it had been obtained positive results after DBS of the subthalamic nucleus of three patients with Parkinson's disease and OCD. CONCLUSION: This article has as target the demonstration that bilateral DBS of the limbic part of the subthalamic nucleus is an alternative for the treatment of refractory OCD. PMID- 17918110 TI - [Zonisamide and neuropathic pain]. AB - INTRODUCTION: In neuropathic pain, as occurs in epilepsy, researchers are striving to find a drug capable of inhibiting the pain-generating ectopic discharges that are produced as a result of neuronal hyperexcitability. This is mediated by ionic exchanges across the channels of the synaptic membrane. This is why the drugs that act on the different types of channels involved in this transmission can regulate neuronal hyperexcitability and therefore have an effect on the pain. DEVELOPMENT: In recent years researchers have gained a deeper understanding of the mechanisms of action of antiepileptic drugs and, since the discovery of their action on one or several synaptic channels, the use of these agents to treat neuropathic pain has become increasingly common. Patients suffering from central pain are also beginning to benefit from the administration of these drugs, especially agents that have proved to be capable of acting with several mechanisms of action and on several channels at the same time. In addition, fewer and less severe side effects are produced, something that is fundamental if we bear in mind the characteristics of patients with central pain, most of whom are adults and elderly. This, together with the fact that there are fewer interactions with other drugs, has led to the new antiepileptic drugs' becoming the preferred medication for this pathology today. CONCLUSIONS: Zonisamide acts on several types of channels and it is known to have four different mechanisms of action, which means it can be effective in treating these patients, although further studies are required (above all randomised double blind trials) in order to really evaluate the usefulness of these drugs in the treatment of neuropathic pain. PMID- 17918111 TI - [Lumbar puncture: its indications, contraindications, complications and technique]. AB - INTRODUCTION: Although first described over 100 years ago, lumbar puncture is still an important tool in the diagnosis of neurological diseases. In this article we review its indications, contraindications, the technique for carrying it out, the analysis of the cerebrospinal fluid and possible complications. DEVELOPMENT: The lumbar puncture has diagnostic and therapeutic indications. The chief diagnostic indications include infectious, inflammatory and neoplastic diseases affecting the central nervous system. Complications are infrequent, except for headaches and low back pain, but can be severe. Analysis of the cerebrospinal fluid must include a cell count and determination of the glucose and protein concentrations. The other analytical studies of cerebrospinal fluid must be conducted according to the diagnostic suspicion. CONCLUSION: The lumbar puncture in expert hands is a safe test. The health professional should be suitably familiar with its contraindications, the regional anatomy and the technique used to perform it. PMID- 17918112 TI - [Experimental models of Huntington's disease]. AB - INTRODUCTION: Huntington's disease (HD) is an autosomal dominant hereditary disease caused by triplet repetition in exon 1 of the huntingtin protein located in chromosome 4. Medium spiny neurons in the striatum are selectively affected. Clinical manifestations include progressive behavioural, motor and cognitive disorders. There is no treatment available today capable of modifying the natural course of the disease. A great amount of research work is being carried out, much of which involves animal models of the disease. DEVELOPMENT: We reviewed the articles published in PubMed on basic research into HD and analysed the most frequently used models. Transgenic mouse models, excitotoxic models, transgenic fly models and cell cultures are all used in studies into HD. The advantages and disadvantages of each of them are highlighted. CONCLUSION: The contribution made by each model of HD must be known in order to draw up a correct design in experimental studies of the disease. PMID- 17918113 TI - [Rapidly progressing cognitive impairment as the presenting symptom of a bithalamic tumour]. PMID- 17918114 TI - [Acute encephalopathy secondary to treatment with disulfiram]. PMID- 17918115 TI - [The FAST study fails to demonstrate the effectiveness of recombinant factor VIIa in intracerebral haemorrhage]. PMID- 17918116 TI - [Cuban neurosciences from a gender perspective]. PMID- 17918118 TI - Bacterial gangs attack implants. PMID- 17918120 TI - Microbiology of infection in mechanical circulatory support. AB - The results of MCSS therapy for patients with severe heart failure are improving, and there are increasing numbers of implants as device therapy enters the era of permanent use (i.e., destination therapy). Device-related infection of implanted pumps and sepsis remain important risk factors for death, and once infections are established on biomaterial surfaces they usually persist despite prolonged antimicrobial therapy. This paper covers the topic of infection in mechanical circulatory support. Methods to prevent and manage infection are described. Recent information from the field of microbiology that is relevant to understanding device-related infection is reviewed, including the mechanisms for microbial adhesion to prosthetic surfaces, quorum sensing, and biofilm formation. Characteristics that are important to antimicrobial resistance of microbes dwelling in biofilms are described (e.g. responses to microenvironmental conditions that result in resistance to antibiotics). This information may lead to new therapies that further diminish rates of device infection, and allow complete eradication of infections when they occur. PMID- 17918119 TI - Bacterial communications in implant infections: a target for an intelligence war. AB - The status of population density is communicated among bacteria by specific secreted molecules, called pheromones or autoinducers, and the control mechanism is called ""quorum-sensing"". Quorum-sensing systems regulate the expression of a panel of genes, allowing bacteria to adapt to modified environmental conditions at a high density of population. The two known different quorum systems are described as the LuxR-LuxI system in gram-negative bacteria, which uses an N-acyl homoserine lactone (AHL) as signal, and the agr system in gram-positive bacteria, which uses a peptide-tiolactone as signal and the RNAIII as effector molecules. Both in gram-negative and in gram-positive bacteria, quorum-sensing systems regulate the expression of adhesion mechanisms (biofilm and adhesins) and virulence factors (toxins and exoenzymes) depending on population cell density. In gram-negative Pseudomonas aeruginosa, analogs of signaling molecules such as furanone analogs, are effective in attenuating bacterial virulence and controlling bacterial infections. In grampositive Staphylococcus aureus, the quorum-sensing RNAIII-inhibiting peptide (RIP), tested in vitro and in animal infection models, has been proved to inhibit virulence and prevent infections. Attenuation of bacterial virulence by quorum-sensing inhibitors, rather than by bactericidal or bacteriostatic drugs, is a highly attractive concept because these antibacterial agents are less likely to induce the development of bacterial resistance. PMID- 17918121 TI - Peritoneal dialysis-related peritonitis: can we predict it? AB - Peritonitis complicating peritoneal dialysis represents a major cause of technique failure, hospitalization, and increased mortality. Peritonitis tends to be recurrent and clustered within particular patients at risk. The aim of this review is to evaluate the potential predictive factors for development of peritoneal dialysis-associated peritonitis based on currently available evidence. Risk factors were divided into medical and non-medical ones, and characterized by a schema of fixed versus modifiable factors. A new direction in the landscape change of the risk factors of peritonitis appears to focus on psychosocial aspects and patient training. Identification of these factors have important clinical implications because of the hitherto lack of well-established strategies to prevent peritonitis complicating peritoneal dialysis. It is hoped that better understanding of the risk factors will allow us to take tangible steps toward minimizing the infectious burden from the Achilles' heel of peritoneal dialysis. PMID- 17918122 TI - Small-colony variants (SCVs) of staphylococci: a role in foreign body-associated infections. AB - Staphylococci have various strategies for resisting therapy that extend beyond classic mechanisms. Clinical experience with device-associated infections as well as with infections due to small-colony variants (SCVs) clearly shows that both antibacterial chemotherapy and host defense mechanisms are often unable to eliminate the pathogens and cure these infections. Of particular interest is the fact that in the past few years an increasing number of various foreign body related infections due to staphylococcal SCVs have been reported. In this overview, the characteristics of SCVs recovered from clinical specimens and of defined mutants displaying the SCV phenotype are described. Their slow growth and changing biochemical and physiological features represent a challenge to clinical laboratory personnel, because recovery, identification, as well as susceptibility testing of these variants need particular efforts. In addition, the reduced susceptibility to aminoglycosides and the ability of SCVs to persist intracellularly require specific attention for the treatment of these infections. Thus, special efforts to search for these variants formed by Staphylococcus aureus or by coagulase-negative staphylococci should be considered when an infection is particularly resistant to therapy, persists for a long period or fails to respond to apparently adequate therapy with antimicrobial compounds. PMID- 17918123 TI - Underestimated collateral effects of antibiotic therapy in prosthesis-associated bacterial infections. AB - Antibiotic treatment of infections associated with the use of indwelling medical devices in ageing and/or severely ill patients represents a significant healthcare problem due to the difficulty of treating such infections and to the various collateral effects that may be observed following the often aggressive therapy. We summarize some effects of antibiotics on the expression of virulence factors of the microorganisms which cause such infections. These effects, particularly those resulting in a stimulation of bacterial virulence, might be usefully included among the other well-known collateral effects of antibiotic therapy. PMID- 17918124 TI - Prevalence of genes encoding for staphylococcal leukocidal toxins among clinical isolates of Staphylococcus aureus from implant orthopedic infections. AB - Staphylococcus aureus has emerged as a major cause of implant infections. It is known that it is able to produce several toxins that contribute to its armory of virulent weapons, but there are still no data on their prevalence among isolates recovered from biomaterial-centered infections. In this study, 200 Staphylococcus aureus isolates from infections related to different types of orthopedic implants (hip and knee arthroprostheses, internal and external fixation devices) were tested by polymerase chain reaction for the prevalence of genes encoding for leukotoxins. Although almost all isolates were positive for the a-hemolysin gene (99%), none was positive for lukM. The leukotoxin genes lukE/lukD were found in 67% of isolates. The presence of lukE/lukD was significantly associated with that of Accessory Gene Regulatory locus agr II. The lukE/lukD-positive isolates were significantly more prevalent in the staphylococcal isolates from knee arthroprostheses than in the isolates from the other implant types. The genes encoding Panton-Valentine leukocidin components were detected in only one isolate that, curiously enough, was taken solely from a knee arthroprosthesis infection. PMID- 17918125 TI - Effects of tigecycline, linezolid and vancomycin on biofilms of viridans streptococci isolates from patients with endocarditis. AB - BACKGROUND: Endocarditis, and prosthetic valve endocarditis in particular, is a serious disease with high morbidity and mortality. We investigate the effects of tigecycline, linezolid and vancomycin on biofilms of viridans group streptococci (VGS) isolated from patients with definite native or prosthetic valve endocarditis. METHODS AND RESULTS: Ten of 20 VGS blood stream isolates from patients with endocarditis formed biofilms in the microtiter plate biofilm model. The minimal inhibitory concentrations (MIC) for tigecycline, linezolid and vancomycin were determined using the microdilution broth method. Biofilms were grown for 24 hours and were incubated with tigecycline, linezolid and vancomycin at increasing concentrations from 1-128x MIC of the isolate being tested. Biofilm thickness was quantified by measuring the optical density (OD) after dyeing it with crystal violet. The incubation of the biofilms with tigecycline, linezolid or vancomycin resulted in a significant reduction of OD compared to the control biofilm without antibiotic (p<0.05). The optical density ratio (Odr) decreased significantly at 2x MIC for tigecycline, and at 8x MIC for linezolid and vancomycin (p<0.05). Although biofilms persisted even at the highest antibiotic concentrations of 128x MIC, bacterial growth was eradicated starting at concentrations of 16x MIC for vancomycin and of 32x MIC for linezolid, but not for tigecycline, up to a concentration of 128x MIC. CONCLUSIONS: In the present study on viridans streptococci isolated from patients with endocarditis, tigecycline and linezolid reduced the density of the biofilms as effectively as vancomycin. However, linezolid and vancomycin were bactericidal at higher concentrations. Linezolid and vancomycin at very high doses may be useful in the treatment of biofilm-associated diseases caused by VGS infections. PMID- 17918126 TI - Biotechnological war against biofilms. Could phages mean the end of device related infections? AB - Microorganisms universally attach to surfaces, resulting in biofilm formation. These biofilms entail a serious problem in daily clinical practice because of the great prevalence of implantable device-related infections. Differences in antibiotic activity against planktonic and sessile bacteria may relate to clinical failures in the treatment of biofilm-related infections (BRI). Bacteriophages have several characteristics that make them potentially attractive therapeutic agents in some selected clinical settings, like for example BRI. They are highly specific and very effective in lysing targeted bacteria, moreover, they appear to be safe for humans. Many studies have shown the potential of phages for the treatment of infectious diseases in plants and animals, including infections with highly drug-resistant bacteria. The therapeutic use of bacteriophages, possibly in combination with antibiotics, may be a valuable approach in BRI. However, many important questions still remain that must be addressed before phages can be endorsed for therapeutic use in humans. PMID- 17918127 TI - Staphylococcus biofilm components as targets for vaccines and drugs. AB - Staphylococci have become the most common cause of nosocomial infections, especially in patients with predisposing factors such as indwelling or implanted foreign polymer bodies. The pathogenesis of foreign-body associated infections with S.aureus and S. epidermidis is mainly related to the ability of these bacteria to form thick, adherent multilayered biofilms. In a biofilm, staphylococci are protected against antibiotic treatment and attack from the immune system, thus making eradication of the infections problematic. This necessitates the discovery of novel prophylactic and therapeutic strategies to treat these infections. In this review, we provide an overview of staphylococcal biofilm components and discuss new possible approaches to controlling these persistent biofilm-dwelling bacteria. PMID- 17918128 TI - Antimicrobial coating of devices for prevention of infection: principles and protection. AB - Device-associated infections are responsible for about half of nosocomial infections and can cause major medical and economical sequelae. Despite adherence to basic infection control measures, which constitute the mainstay for preventing infection, infections associated with certain devices continue to exist at unacceptably high rates. Potentially-preventive, antimicrobial-utilizing strategies include systemic antibiotic prophylaxis and local administration of antimicrobial agents (antibiotics or antiseptics), which includes antimicrobial irrigation of the surgical field, placement of antimicrobial carriers, antiseptic cleansing of the skin, dipping of surgical implants in antimicrobial solutions, and inserting antimicrobial-coated implants. Since bacterial colonization of the indwelling device is a prelude to infection, prevention of device colonization may lead to a lower rate of clinical infection. Different approaches for antimicrobial coating of devices have been variably successful in preventing device-associated infections. Optimal characteristics of antimicrobial coating can help predict the likelihood and degree of clinical protection against infection. This review addresses the impact of device-related infection, antimicrobial-utilizing approaches for preventing infection, clinical protection afforded by different types of antimicrobial coating, characteristics that predict the ability of antimicrobial coating of devices to prevent clinical infection, and future directions of antimicrobial coating. PMID- 17918129 TI - Biomaterial strategies to reduce implant-associated infections. AB - Although the prophylaxis in controlling sterility within the operating room environment has been greatly improved, implant-associated infection is still one of the most serious complications in implant surgeries due to the existence of immune depression in the peri-implant area. The antibacterial ability of materials themselves logically becomes an important factor in preventing implant associated infections. With the understanding of the pathogenesis of implant associated infections, many approaches have been developed through providing an anti-adhesive surface, delivering antibacterial agents to disrupt cell-cell communication and preventing bacteria aggregation or biofilm formation, or killing bacteria directly (lysing the cell membrane). In this article, we review the current strategies in improving the antibacterial ability of materials to prevent implant infection and further present promising tactics in materials design and applications. PMID- 17918130 TI - Innovative methods of rapid bacterial quantification and applicability in diagnostics and in implant materials assessment. AB - In recent years, a variety of new technologies have been proposed that allow rapid qualitative and quantitative microbiological analyses. In this paper we discuss the urgent needs for reliable and rapid microbiological analytical techniques in different applicative fields involving the research, production and medical application of implant materials, and the potential benefits derived from the use of new methods for rapid bacterial quantification. Current compendial methods are easy to perform and have gained confidence over their long period of use, but the supplemental use of new technologies could represent real breakthroughs whenever sensitive and rapid responses are urgently required and not met by the tests currently in use. Overall, the new microbiological methods require critical evaluation depending on their specific type of application and they may still not be thought of as totally substitutive, but they certainly exhibit considerable potential for different areas of biomaterials, as well as for advanced therapy medicinal and tissue engineering treatments. PMID- 17918131 TI - Cave enterococcum! PMID- 17918132 TI - Access to kidney transplantation: the limitations of our current understanding. AB - Since kidney transplantation (KTX) is the preferred means of treating kidney failure, ensuring that all patients who may benefit from KTX have equal access to this scarce resource is an important objective. Studies focusing on this issue will become increasingly important as the gap between the demand and supply of organs continues to increase, and changes to the United Network of Organ Sharing organ allocation policy are actively debated. However, it is clear that current methods used to study access to KTX have serious limitations. This review highlights the shortcomings of the methods currently used to assess access to KTX, and the limitations of registry data and national wait-list data as information sources to study patient access to KTX. The review also provides suggestions for research and analytical approaches that might be utilized to improve our future understanding of patient access to KTX. The information provided will aid the reader to critically assess issues related to patient access to KTX. PMID- 17918133 TI - Disorders of renal magnesium handling explain renal magnesium transport. AB - Magnesium is essential for bone stability, neuronal excitability, muscular relaxation and many other metabolic functions. Despite its fundamental biological importance, mechanisms controlling systemic magnesium homeostasis are only partially understood. The kidney plays a central role in maintaining magnesium balance as evident from several rare inherited disorders of renal magnesium transport. Recent studies shed new light on molecular mechanisms of renal magnesium handling and its control. Mutations in the claudin 16 (paracellin) paracellular protein in the thick ascending limb (TAL) of Henle's loop and in the transient receptor potential cation channel, subfamily 6, member 6 (TRPM6) magnesium channel expressed in distal tubules found in patients with renal magnesium wasting and hypomagnesemia underscore the importance of these transport proteins. A study by Hou et al (J Biol Chem 2007; 282: 17114-22) demonstrates a pathomechanism for claudin 16 mutations that gives interesting insights into the function of the TAL. Moreover, Groenestege and colleagues report (J Clin Invest 2007; 117: 2260-7) the identification of the epidermal growth factor (EGF) as a hormonal regulator of TRPM6 activity, and thereby explain how mutations in EGF can cause familial hypomagnesemia. Interestingly, cetuximab, a drug used in treatment of certain cancers, acts an inhibitor of the EGF receptor and causes hypomagnesemia which may be due to the inhibition of EGF signaling. PMID- 17918134 TI - Focus on vitamin D therapy in chronic kidney disease. PMID- 17918135 TI - From Zanius to ADMA: ADMA - a new ''adipocytokine'' and its potential role in metabolic syndrome. PMID- 17918136 TI - A perspective on arterionephrosclerosis: from pathology to potential pathogenesis. AB - The morphological findings of so-called hypertensive nephrosclerosis, rather than implying a linear direct relationship to damage induced by hypertension, may indicate complex environmental and genetic factors, which together foster the coexistence of renal lesion and hypertension in this clinical setting. We discuss the clinical and pathological criteria for diagnosis of arterionephrosclerosis, and possible pathogenetic factors, including hypertension, ethnicity, aberrant autoregulation, prothrombotic mechanisms, low birth weight, decreased nephron number, genetic factors and dysmetabolic syndrome. PMID- 17918137 TI - Effectiveness and safety of different hemodialysis modalities: a review. AB - BACKGROUND AND AIMS: At present, there are a number of hemodialysis (HD) modalities that appear to be superior to conventional HD. This study sought to compare and assess the effectiveness and safety of the various hemodialysis modalities. METHODS: REVIEW OF THE LITERATURE: multiple electronic databases were searched, including Medline, EMBASE and Cochrane Collaboration. Papers were independently selected by 2 researchers. RESULTS: Thirty-five original papers were included. The outcomes of the largest randomized clinical trial undertaken (the Hemodialysis [HEMO] Study) showed that high- versus low-flux HD did not significantly reduce overall mortality (p=0.23) but did reduce the risk of hospitalization and death due to vascular causes by 10% (p<0.05). The results of the respective studies are controversial as to the benefits of high-flux HD on anemia, nutritional status or cardiovascular risk factors. The only study that provided mortality results for hemodiafiltration failed to observe significant differences between high- and low-flux HD. Similarly, there was no clear evidence of improvements in any of the other outcomes investigated. CONCLUSIONS: The results of this review do not allow for any conclusion to be drawn as to which HD modality is best in terms of mortality, morbidity and patients' quality of life. The different HD techniques do not show major improvements in any of the outcomes assessed, but these cannot be dismissed since the studies had multiple flaws and lacked long-term follow-up data. Better-quality studies are needed to establish which hemodialysis modalities are best suited to individual patients' specific clinical conditions. PMID- 17918138 TI - The QUEST initiative anemia study in transplant patients: rationale and study protocol. PMID- 17918139 TI - The QUEST initiative anemia study in ESRD: rationale and study protocol. PMID- 17918140 TI - The enzymatic machinery for ADMA synthesis and degradation is fully expressed in human adipocytes. AB - BACKGROUND: The endogenous inhibitor of nitric oxide synthase (NOS), asymmetric dimethylarginine (ADMA), is implicated in endothelial dysfunction and is a marker of renal disease progression and cardiovascular (CV) complications. Various cell species exhibit the enzymatic system that generates and degrades this methylarginine, but it is unknown whether this machinery is expressed in adipocytes. The question is relevant because adipocyte-derived mediators are implicated both in renal and cardiovascular diseases. METHODS: We measured ADMA concentration in pure adipocytes in culture and measured mRNA levels of the enzymes involved in ADMA metabolism (real-time polymerase chain reaction) both in pure adipocytes in culture and in adipose tissue harvested in 9 healthy subjects. These enzymes included protein arginine N-methyltransferases type I (PRMTs) involved in ADMA synthesis, dimethylarginine dimethylaminohydrolases (DDAHs) responsible for ADMA degradation and constitutive and inducible forms of NOS (i.e., NOS1, NOS2A and NOS3 genes), the main functional target of ADMA. RESULTS: Human adipocytes express the whole gene set that codes for the enzymatic system responsible for the biosynthesis and the degradation of ADMA, and this methylarginine is actually released by adipocytes in culture. NOS gene isoforms have a low level of expression in human adipose tissue, indicating that putative functions of ADMA in fat cells may be in part mediated by mechanisms other than NOS inhibition. CONCLUSIONS: Human adipocytes produce ADMA and express the full enzymatic machinery responsible for ADMA metabolism. Studying the functional implication of these findings may be of relevance for clarifying the role of fat mass expansion in human disease. PMID- 17918141 TI - Impaired antiviral activity of monocytes from patients on hemodiafiltration. AB - BACKGROUND: The aim of our study was to determine whether intermittent hemodiafiltration (HDF) leads to an alteration in monocyte antiviral activity as well as in the in vitro release of cytokines such as interleukin-12 (IL-12), tumor necrosis factor-alpha (TNF-alpha) and interferon-alpha (IFN-alpha) by the same cells. METHODS: We enrolled 25 patients undergoing HDF for 3.5-4 hours 3 times a week (12 men, 13 women; mean age 58 +/- 6.7 years) and 25 healthy donors (ND) (12 men, 13 women; mean age 57 +/- 8 years). Monocytes from peripheral blood mononuclear cells were isolated with a Monocyte Isolation Kit II. Monocytic cells were infected with herpes simplex virus type 2 (HSV-2). Cytokines were assayed in supernatants. RESULTS: The in vitro antiviral activity of monocytes from HDF patients was significantly impaired with respect to ND. Furthermore, monocytes from post-HDF patients were more prone to viral infection. Lipopolysaccharide (LPS) stimulation induced significant viral inhibition only in monocytes from NDs (p<0.05). The cytokine pattern (TNF-alpha, IFN-alpha and IL-12) in monocytes stimulated with LPS was markedly inhibited in HDF patients compared with ND (p<0.05). A basal production of TNF-alpha was found in monocytes from pre-HDF and post-HDF patients. No IFN-alpha production was found in LPS-stimulated and HSV-2 infected monocytes from pre-HDF and post-HDF patients. IL-12 production appeared significantly decreased after HDF in all experimental conditions (p<0.05). CONCLUSIONS: The significant increase of viral replication in monocytes from HDF patients compared with healthy donors could be related to a significant reduction of cytokine production. Moreover, the dialytic session influenced the intrinsic antiviral activity of monocytes, favoring viral replication. PMID- 17918142 TI - A complete genomic analysis of the apolipoprotein E gene in Chinese patients with lipoprotein glomerulopathy. AB - Mutations of the apolipoprotein E (apo E) gene are thought to play an etiological role in lipoprotein glomerulopathy (LPG), a novel kidney disease. Evidence suggesting that mutated forms of apo E may be present in LPG includes elevated plasma concentrations of apo E in LPG patients and deposition of apo E in their glomerular capillaries, and there are published reports indicating that mutations of the apo E gene are present in the Japanese LPG patient population. Conflicting reports, however, exist in the current literature. Therefore, to strengthen the theory that apo E gene mutations may be present in LPG pathogenesis, we chose to examine the 5.5 kb genomic DNA encompassing the entire apo E locus and adjoining flanking regions, in 17 Chinese LPG patients and 10 normal controls. Results suggest that there are no apo E gene mutations present in our LPG patients, including previously reported apo E mutations associated with LPG. Surprisingly, however, when the apo E sequence used in the current study was compared against the apo E gene sequence published in GenBank, 10 ethnic variation sites were identified in the noncoding regions. We therefore conclude that apo E gene mutations may not be the only etiology of LPG, and it is likely that other abnormalities could play a role in the pathogenesis of the disease. PMID- 17918143 TI - A modified Cockcroft-Gault formula taking into account the body surface area gives a more accurate estimation of the glomerular filtration rate. AB - We sought to validate the improvement by adjustment for body surface area (BSA) of the accuracy of the original Cockcroft-Gault equation to estimate glomerular filtration rate (GFR), in a prospective cross-sectional study of 269 European patients with chronic kidney disease (CKD). We compared 3 methods: original Cockcroft-Gault equation, modified Cockcroft-Gault formula adjusted for BSA and abbreviated Modification of Diet in Renal Disease (MDRD) equation, using inulin clearance. Statistical analyses comprised repeated-measures analysis of variance (ANOVA), determination of the Pearson coefficient of correlation and a Bland Altman concordance study. The ability of the GFR estimates to properly categorize patients in K/DOQI stages of CKD was also examined. Inulin clearance differed significantly from the standard Cockcroft-Gault method (ANOVA, p<0.001) and the abbreviated MDRD method (ANOVA, p<0.001) but not from the BSA-modified Cockcroft Gault formula. Inulin clearance correlated better with the BSA-modified Cockcroft Gault formula (r=0.88) and abbreviated MDRD equation (r=0.87) than with the standard Cockcroft-Gault equation (r=0.82). In concordance studies, bias was far smaller with the BSA-modified Cockcroft-Gault formula (mean bias -1.75 ml/min), than with the standard Cockcroft-Gault equation (mean bias -4.72 ml/min). The bias of the abbreviated MDRD was larger (mean bias +6.24 ml/min). Only patients with CKD stage 1 were better categorized with the BSA-modified Cockcroft-Gault formula and with the standard Cockcroft-Gault estimate than with the abbreviated MDRD equation. We conclude that adjustment for body surface area improves accuracy of the original Cockcroft-Gault equation. PMID- 17918144 TI - Physiological similarities and differences between renal aging and chronic renal disease. AB - Even though there are some functional similarities between the aged kidney and the chronically damaged one, such as the reduction in glomerular filtration rate and in the sodium-water reabsorption capability, there are many physiological differences between these two groups, as is the case of erythropoietin, urea, potassium, calcium, phosphorus and magnesium renal handling. Thus, the data presented demonstrate that renal aging and chronic kidney disease constitute different clinical scenes. PMID- 17918145 TI - Thrombospondin-1 mRNA expression in experimental kidney transplantation with heart-beating and non-heart-beating donors. AB - BACKGROUND: Ischemia-reperfusion (IR) is a risk factor for delayed graft function, a clinical syndrome more frequently observed in non-heart-beating donors (NHBDs). Previous studies have reported that transforming growth factor beta1 (TGF-beta1) and hypoxia-inducible factor-1alpha (HIF-1alpha) gene expression increase in the first few days after kidney transplant and that this increase in TGF-beta1 expression is lower in NHBD animals. The purpose of this study was to extend the gene profile analysis by characterizing TGF-beta1 activator thrombospondin-1 (TSP-1) and genes related to HIF-1alpha such as heme oxygenase-1 (HO-1), nitric oxide synthase-2 (NOS-2) and NOS-3. METHODS: The experimental pig model of kidney transplantation comprised heart-beating donors (HBDs, n=9) and NHBDs (n=22). Cortical biopsies were collected after anesthetic induction (baseline), after warm ischemia (WI), after cold ischemia (CI), after 1 hour of reperfusion (1R) and 5 days (5D) after transplant. TSP-1, HO-1, NOS-2 and NOS-3 mRNA expression was determined by real-time PCR. RESULTS: No change in expression of any of the genes analyzed was found during the transplant procedure (WI, CI, 1R) in HBD and NHBD cortical samples. TSP-1 mRNA was significantly increased at 5D in NHBD animals but unchanged in the HBD group. HO-1 was up regulated in HBD (p<0.05) and NOS-2 mRNA was significantly increased in both groups (p<0.05). No difference in NOS-3 expression was observed at 5D. CONCLUSIONS: The increased TSP-1 expression in NHBDs may indicate a compensatory response to the reported diminished TGF-beta1 expression. The augmented NOS-2 and HO-1 expression in HBDs could have a positive effect on the recovery of kidney function. PMID- 17918146 TI - Comparative immunogenicity of 2 recombinant hepatitis B vaccines (GeneVac-B and Engerix-B) in adult patients with chronic renal failure. AB - BACKGROUND: Hepatitis B virus infection is often fatal or results in a carrier state with uremia. Vaccination protects against infection, but immune response may remain low. We aimed to study immune responses to 2 recombinant hepatitis B vaccines (Engerix-B and GeneVac-B) in patients with end-stage chronic renal failure. METHODS: Patients initially negative for hepatitis B (HB) surface antigen (HBsAg), and with core antibody (anti-HBc) and nonprotective anti-HBs titers (<100 mIU/mL) were selected. Four doses of 40 microng of either vaccine were administered intramuscularly, further doses being given 1, 2 and 6 months after the first dose. Seroconversion and seroprotection were defined as anti-HBs level above 10 mIU/mL and 100 mIU/mL, respectively. RESULTS: A total of 11 and 9 subjects receiving GeneVac-B and Engerix-B, respectively, completed the protocol. Seroconversion was achieved in 100% for both vaccines. Seroprotection occurred in 78% and 82% of the Engerix-B and the GeneVac-B recipients, respectively. Geometric mean titers 1 month after the fourth dose were 274 mIU/mL (95% confidence interval [95% CI], 71-1,057 mIU/mL) and 322 mIU/mL (95% CI, 142-730 mIU/mL), respectively. CONCLUSIONS: Both vaccines are highly immunogenic in renal failure and did not show significant differences between each other. PMID- 17918147 TI - Regional citrate versus heparin anticoagulation during venovenous hemofiltration in patients at low risk for bleeding: similar hemofilter survival but significantly less bleeding. AB - BACKGROUND: Regional citrate anticoagulation (RCA) may be a valuable alternative to systemic heparin anticoagulation during continuous venovenous hemodialysis (CVVH). RCA does not increase the risk of bleeding, but carries the risk of metabolic derangements. Clotting-free circuit survival may be increased with RCA. METHODS: Critically ill patients at low risk for bleeding were randomized for CVVH with RCA or systemic heparin anticoagulation. A maximum of 4 consecutive CVVH circuits were analyzed for every patient. Clotting-free circuit survival, adverse metabolic events and bleeding complications were recorded. RESULTS: Forty eight patients were included in the study and randomized for RCA (n=21) or systemic heparin (n=27). A total of 142 CVVH circuits were analyzed with a median of 3 circuits per patient. Uncensored circuit survival and median clotting-free circuit survival were similar for both groups. No significant adverse metabolic events occurred in the RCA group. Major bleedings were not recorded in the RCA group, compared with 10 events in the heparin group (p<0.01). The number of red blood cell units given per day of CVVH was significantly higher in the heparin group (0.88 vs 0.43 units/day, p=0.01). Also, the mean decrease in hemoglobin concentration per day in the nontransfused stable patients, was significantly higher in the heparin group. CONCLUSION: RCA during CVVH can be performed safely, with a circuit survival similar to systemic heparin anticoagulation, but significantly lower incidence of bleeding. Citrate anticoagulation should be advocated as first choice, also for critically ill patients considered to be at low risk for bleeding. PMID- 17918149 TI - Renal function and functional reserve in healthy elderly individuals. AB - BACKGROUND: Aging is characterized by a decline in renal function and by a susceptibility to renal diseases. However it is not clear whether the observed changes are solely hemodynamic, structural or both. We evaluated renal function, functional reserve (RFR) and morphology in healthy elderly individuals. METHODS: Healthy participants (n=19) were divided into young (n=6, age range 25-37 years), middle-aged (n=6, 44-74 years) and elderly (n=7, 81-96 years). Nitric oxide (NO), plasma renin activity (PRA) and aldosterone, renal plasma flow (RPF) by p aminohippurate clearance (CPAH) and glomerular filtration rate (GFR) by inulin clearance (CIN) were determined before and during maximal vasodilating stimuli, induced with the infusion of dopamine and amino acids. Glomerular sclerosis, lumen area and wall thickness of afferent arterioles were determined by kidney biopsy from 36 healthy kidney donors and from 6 nephrectomies for renal carcinoma. RESULTS: GFR and RPF were slightly reduced in elderly individuals whereas filtration fraction (FF) was increased. GFR and RPF did not increase in the elderly after maximal vasodilating stimuli as in young and middle-aged subjects suggesting a reduction of RFR. NO, increased at baseline, did not increase further after vasodilating stimuli; while on the contrary, PRA, similar in the 3 groups at baseline, was not reduced by vasodilating stimuli in the elderly. Sclerotic glomeruli but not glomerular volume were significantly increased by aging. Afferent arteriole lumens were reduced by aging whereas wall thickness was unchanged. CONCLUSIONS: Renal function is preserved with aging in healthy subjects at the expense of a complete reduction of RFR. RFR may be wasted to compensate for the increased number of sclerotic glomeruli. Vascular changes, suggested by reduced arteriolar lumen, may be so advanced that even in the presence of high levels of vasodilatory molecules, kidneys are not responsive anymore to maximal vasoactive stimuli. PMID- 17918148 TI - Similar HCV viral load levels and genotype distribution among end-stage renal disease patients on hemodialysis and HCV-infected patients with normal renal function. AB - BACKGROUND: At present it is not clearly established if hepatitis C virus (HCV) RNA levels and genotype distribution have any peculiar aspect in HCV-infected end stage renal disease (ESRD) patients. The aim of this study was to evaluate in HCV infected ESRD patients, the alanine aminotransferase (ALT) profile, HCV-RNA levels and genotype distribution, comparing them with HCV-infected patients with normal renal function. METHODS: A cross-sectional study was performed in 66 hemodialysis patients (group 1) and 264 subjects with normal renal function (group 2). All participants in both groups had detectable HCV-RNA. Mean ALT levels were determined in all subjects as well the viral load and the genotype. RESULTS: Groups were similar according to gender and age. ALT was normal in 74% patients in group 1 and in 23% in group 2 (p<0.001). The median viral load was 5.3 x 10(5) IU/mL in group 1 and 6.6 x 10(5) IU/mL in group 2 (p=0.23). Genotype 1b was the most prevalent in both groups (56% vs. 57%; p=0.38). CONCLUSION: HCV infected ESRD patients have lower ALT levels, but the viral load and the genotype distribution are similar to those observed in HCV-infected individuals with normal renal function. PMID- 17918151 TI - Vascular access-related thrombotic complications: research hypotheses and therapeutic strategies. PMID- 17918150 TI - Focal segmental glomerulosclerosis due to a relapsing non-Hodgkin's lymphoma diagnosed by positron emission tomography. AB - Glomerulonephritis rarely appears associated with Hodgkin's disease or non Hodgkin's lymphoma (NHL). We present a patient with a relapse of a non-Hodgkin's lymphoma which first presented as nephrotic syndrome due to focal segmental glomerulosclerosis (FSGS). This case report discusses the unusual association of non-Hodgkin's lymphoma and focal segmental glomerulosclerosis, as well as the crucial role of positron emission tomography in detecting the relapsing lymphoma. PMID- 17918152 TI - Cigarette smoking and risk of borderline and invasive epithelial ovarian cancer. AB - Studies regarding the association between smoking and risk of epithelial ovarian cancer (EOC) are inconsistent. The purpose of this study was to examine the association between smoking and EOC, overall and according to invasiveness and histological subtype in a cohort of women with a high proportion of smokers at enrollment. We followed 103,081 women, aged 30-50 years in 1991/1992, from the Norwegian-Swedish Women's Lifestyle and Health cohort. The women completed a questionnaire on personal characteristics and exposures at enrollment and were subsequently followed with linkages to national registers through December 31, 2004. We used Cox proportional hazard regression models to estimate hazard ratio (RR) of EOC with 95% confidence intervals (CIs) associated with different measures of smoking exposures adjusting for confounding variables. Altogether 343 [241 (70%) invasive and 102(30%) borderline] incident EOC cases were identified. Former [HR = 2.2(95% CI 1.0-4.7)] and current [HR = 2.7(95% CI 1.2-5.7)] smokers had a more than doubling in risk for borderline tumors compared to never smokers. Women who had smoked for more than 20 years had 3 times [HR = 3.1(95% CI 1.5 6.7)] the risk of borderline tumors compared to never smokers. A test for trend according to smoking status was almost significant for mucinous tumors (p-trend = 0.05). A significant dose response relationship was found according to smoking intensity [pack-years; (0-9, 0-14, >or= 15)] and duration [number of years; (0 10, 11-20, >or= 20)] for borderline and serous tumors (p-trends < 0.05). In conclusion, smoking may increase the risk of borderline EOC. PMID- 17918153 TI - Colon carcinogenesis in liver-specific IGF-I-deficient (LID) mice. AB - Circulating insulin-like growth factor I (IGF-I) is associated with increased risk of colorectal cancer. It is not clear, however, whether IGF-1 plays a direct causative role in colon carcinogenesis or whether it mediates the known promoting effects of insulin. The objective of this study was to determine the role of IGF 1 in colon carcinogenesis using liver-specific IGF-I deficient (LID) mice that exhibit 70% reductions in circulating IGF-I. Female and male LID mice were treated with the colon-specific carcinogen azoxymethane to induce aberrant crypt foci (ACF) or colon tumors. Female LID mice developed significantly fewer ACF and had normal insulin levels compared to controls. Male LID mice, however, were hyperinsulinemic and exhibited no significant differences in ACF development compared to controls. In the tumor study, both male and female LID mice were hyperinsulinemic and had no significant differences in tumor incidence or multiplicity compared to their respective controls. There was a significant 25% reduction in tumor size, however, in both male and female LID mice compared to controls. These data suggest that IGF-I deficiency attenuates the promoting effect of insulin on colon carcinogenesis and that IGF-I is an independent promoter of the growth of established tumors. Our findings implicate both IGF-I and insulin as important promoters of colon cancer development. PMID- 17918154 TI - Germline mutations in the CHEK2 kinase gene are associated with an increased risk of bladder cancer. AB - Germline mutations in CHEK2 have been associated with a range of cancer types but little is known about disease risks conveyed by CHEK2 mutations outside of the context of breast and prostate cancer. To investigate whether CHEK2 mutations confer an increased risk of bladder cancer, we genotyped 416 unselected cases of bladder cancer and 3,313 controls from Poland for 4 founder alleles in the CHEK2 gene, each of which has been associated with an increased risk of cancer at other sites. A CHEK2 mutation (all variants combined) was found in 10.6% of the cancer cases and in 5.9% of the controls (OR = 1.9; 95%CI 1.3-2.7; p = 0.0003). We conclude that CHEK2 mutations increase the risk of bladder cancer in the population. PMID- 17918155 TI - The role of insulin-like growth factor binding protein-3 in the growth inhibitory actions of androgens in LNCaP human prostate cancer cells. AB - Insulin-like growth factor binding protein-3 (IGFBP-3), an antiproliferative and proapoptotic protein, has been shown to be upregulated by growth inhibitory concentrations of androgens in LNCaP human prostate cancer (PCa) cells, but the mechanism of regulation and the role of IGFBP-3 in the modulation of PCa cell proliferation are unknown. In this study, we have examined the effects of a range of concentrations of the synthetic androgen R1881 on IGFBP-3 expression and cell growth in LNCaP cells. We have also investigated the role of androgen-stimulated IGFBP-3 in androgen-induced growth inhibition. We show that low doses of R1881 stimulate LNCaP cell proliferation, but do not induce IGFBP-3 expression, whereas high doses of R1881 that inhibit cell growth, significantly increase expression of IGFBP-3. Importantly, we demonstrate that the combination of calcitriol and androgens not only synergistically upregulates IGFBP-3 expression but also inhibits cell growth better than either hormone alone. siRNA knockdown of IGFBP-3 expression partially reverses the growth inhibition by calcitriol and by androgens. Furthermore, we find that the growth inhibitory dose of R1881 leads to increases in the cyclin dependent kinase inhibitors (CDKIs), p21 and p27 as well as to G1 arrest. These changes can be blocked or partially reversed by IGFBP-3 siRNA, indicating that the induction of CDKIs is downstream of IGFBP-3. Our data suggest, for the first time, that IGFBP-3 is involved in the antiproliferative action of high doses of androgens partly through p21 and p27 pathways and that IGFBP-3 may contribute significantly to androgen-induced changes in LNCaP cell growth. PMID- 17918156 TI - Differential expression of E-prostanoid receptors in human hepatocellular carcinoma. AB - Recent studies have shown that inhibition of cyclooxygenases (e.g. COX-2) exerts antitumorigenic effects on hepatocellular carcinomas (HCCs), which are to a significant extent due to the abrogation of PGE(2) synthesis. PGE(2) acts via differentially regulated prostaglandin receptors (EP(1-4)). Our study was designed to investigate the expression pattern of EP-receptors in HCCs and to evaluate the therapeutic potential of selective EP-receptor antagonists. Using tissue microarrays including a total of 14 control livers, 17 liver cirrhoses, 22 premalignant dysplastic nodules (DNs) and 162 HCCs with different histological grades, the expression of COX-2, mPGES-1 and -2 and EP(1-4)-receptors was analyzed. Western immunoblot analyses were performed to confirm the expression in HCC cell lines. The effects of EP(1-4)-receptor antagonism on cell viability and apoptosis were investigated using MTT-assays and FACS-analyses, respectively. COX 2, mPGES-1 and -2 and EP(1-4)-receptors were expressed in all HCC tissues. COX-2 expression was highest in DNs and declined with loss of HCC-differentiation. With respect to COX-2 expression, a converse expression of EP(1-3) -receptors and mPGES-1 and -2 was found in DNs compared to HCCs. Selectively antagonizing EP(1)- and EP(3)-receptors reduced the viability of HCC cells in a dose-dependent manner, which was associated with apoptosis induction. Our results suggest a differential regulation of EP-receptor subtype expression with dedifferentiation of HCCs in which a converse expression pattern for COX-2 in comparison to EP(1-3) receptors occurs. Of clinical interest, selectively antagonizing EP(1)- and EP(3) receptors may provide a novel systemic therapeutic approach to the treatment of HCCs. PMID- 17918157 TI - Nm23-H1 expression of metastatic tumors in the lymph nodes is a prognostic indicator of oral squamous cell carcinoma. AB - We recently reported that low Nm23-H1 expression of primary oral squamous cell carcinoma (OSCC) was correlated with the occurrence of lymphatic metastasis. However, little is known about whether Nm23-H1 level of metastatic tumors in the cervical lymph nodes is reduced in comparison with primary oral cancers and its significance for patients' prognosis. By immunohistochemistry, we analyzed the Nm23-H1 expression in 52 pairs of OSCC specimens from primary oral cancers and their metastatic lymph nodes. Western blot analysis further confirmed the immunohistochemical interpretation. To verify the effects of Nm23-H1 on cell migration and invasion, we established several stable clones derived from a human OSCC cell line (SAS) by knockdown and overexpression. Wound-healing closure, transwell migration and invasion assays were performed to determine cell motility, migratory and invasive activities. Western blot analysis was carried out to evaluate cyclin A expression of OSCC cells with the altered Nm23-H1 levels following knockdown and overexpression. By immunohistochemistry, Nm23-H1 expression of metastatic lymph nodes was significantly lower than that of their primary oral cancers, supporting a role of Nm23-H1 in metastasis suppression. Negative Nm23-H1 interpretation of OSCC specimens, in either primary oral cancers or metastatic lymph nodes, indicated a poor survival outcome of patients. On the basis of in vitro studies of Nm23-H1 knockdown and overexpression, we demonstrated an inverse correlation between Nm23-H1 expression and the invasiveness of OSCC cells. Moreover, we observed the concomitant reduction in Nm23-H1 and cyclin A levels of metastatic tumors in both results of in vitro OSCC cells and ex vivo tumor specimens. PMID- 17918158 TI - Curcumin enhances the effects of 5-fluorouracil and oxaliplatin in mediating growth inhibition of colon cancer cells by modulating EGFR and IGF-1R. AB - Curcumin (diferuloylmethane), which has been shown to inhibit growth of transformed cells, has no discernible toxicity and achieves high levels in colonic mucosa. 5-fluorouracil (5-FU) or 5-FU plus oxaliplatin (FOLFOX) remains the backbone of colorectal cancer chemotherapeutics, but with limited success. The present investigation was, therefore, undertaken to examine whether curcumin in combination with conventional chemotherapeutic agent(s)/regimen will be a superior therapeutic strategy for colorectal cancer. Indeed, results of our in vitro studies demonstrated that curcumin together with FOLFOX produced a significantly greater inhibition (p < 0.01) of growth and stimulated apoptosis (p < 0.001) of colon cancer HCT-116 and HT-29 cells than that caused by curcumin, 5 FU, curcumin + 5-FU or FOLFOX. These changes were associated with decreased expression and activation (tyrosine phosphorylation) of EGFR, HER-2, HER-3 (72 100%) and IGF-1R (67%) as well as their downstream effectors such as Akt and cycloxygenase-2 (51-97%). Furthermore, while these agents produced a 2-3-fold increase in the expression of IGF-binding protein-3 (IGFBP-3), curcumin together with FOLFOX caused a 5-fold increase in the same, when compared to controls. This in turn led to increased sequestration of IGF by IGFBP-3 rendering IGF-1 unavailable for binding to and activation of IGF-1R. We conclude that the superior effects of the combination therapy of curcumin and FOLFOX are due to attenuation of EGFRs and IGF-1R signaling pathways. We also suggest that inclusion of curcumin to the conventional chemotherapeutic agent(s)/regimen could be an effective therapeutic strategy for colorectal cancer. PMID- 17918160 TI - Dose-dependent dual effect of HTLV-1 tax oncoprotein on p53-dependent nucleotide excision repair in human T-cells. AB - In this study we investigated the effect of Tax on nucleotide excision repair (NER) in human T-cell lines by using the host cell repair analysis of UVC irradiated reporter plasmid. This analysis revealed a p53-dependent NER activity in wild type (w.t.) p53-containing T-cells and p53-independent NER in w.t. p53 lacking T-cells. Notably, in the w.t. p53-containing cells Tax exerted a dose dependent dual effect on NER. While low Tax doses markedly stimulated this repair, high Tax doses strongly reduced it. Further experiments demonstrated that the low Tax doses enhanced, in these cells, the level and the transcriptional function of their w.t. p53 protein. On the other hand, although the high Tax doses further increased the level of p53, they functionally inactivated its accumulating molecules. Both of these Tax effects on p53 proved to be mediated by Tax-induced NF-kappaB-related mechanisms. Together, these data suggest that by NF kappaB activation Tax elevates the level of the cellular w.t. p53. However, while at low Tax doses the elevating w.t. p53 molecules are functionally active and capable of stimulating NER, intensifying further the NF-kappaB activation by the high Tax doses concomitantly evokes certain mechanism(s) which functionally inactivates the accumulating p53 protein. In contrast to this dual effect on the p53-dependent NER, Tax displayed only an inhibitory effect on the p53-independent NER by its high doses, whereas its low doses had no effect on this repair. The mechanisms of the NF-kappaB-associated effects on the level and function of the cellular w.t.p53 and of the p53-independent NER noted in our experimental systems are further investigated in our laboratory. PMID- 17918159 TI - Naftopidil, a selective alpha-1 adrenoceptor antagonist, inhibits growth of human prostate cancer cells by G1 cell cycle arrest. AB - Alpha-1 adrenoceptor antagonists are generally prescribed for benign prostate hyperplasia with lower urinary tract symptoms. Naftopidil, a selective alpha-1 adrenoceptor antagonist, is frequently used in Japan because it has fewer side effects. Here we demonstrate for the first time that naftopidil has growth inhibitory effect in androgen-sensitive and -insensitive human prostate cancer cell lines. The concentrations causing 50% inhibition (IC50) of cancer cell growth were 22.2 +/- 4.0 microM in androgen-sensitive LNCaP cells and 33.2 +/- 1.1 microM in androgen-insensitive PC-3 cells. FACS analysis revealed that cell growth inhibition by naftopidil was due to the arrest of the G1 cell cycle. Expressions of p27(kip1) and p21(cip1) were significantly increased in LNCaP cells treated with naftopidil. In PC-3 cells, naftopidil induced p21(cip1) but not p27(kip1). In vivo, oral administration of naftopidil to nude mice inhibited the growth of PC-3 tumors as compared to vehicle-treated controls. These results suggest that naftopidil may be useful in the chemoprevention of prostate cancer and the intervention of hormone refractory prostate cancer. PMID- 17918162 TI - Garlic (Allium sativum L.): adverse effects and drug interactions in humans. AB - Garlic (Alllium sativum L., Fam Liliaceae) is used medicinally mainly for the treatment of hypercholesterolemia and prevention of arteriosclerosis. Clinical trials have consistently shown that "garlic breath" and body odor are the most common (and well-documented) complaints associated to garlic intake. Case reports have highlighted the possibility that garlic use may cause allergic reactions (allergic contact dermatitis, generalized urticaria, angiedema, pemphigus, anaphylaxis and photoallergy), alteration of platelet function and coagulation (with a possible risk of bleeding), and burns (when fresh garlic is applied on the skin, particularly under occlusive dressings). Consumption of garlic by nursing mothers modifies their infant's behavior during breast-feeding. Finally, garlic may enhance the pharmacological effect of anticoagulants (e. g. warfarin, fluindione) and reduce the efficacy of anti-AIDS drugs (i. e. saquinavir). PMID- 17918161 TI - A comparison of community pharmacists' views of over-the-counter omeprazole and simvastatin. AB - PURPOSE: Simvastatin and omeprazole were re-classified to pharmacy status in Great Britain in summer 2004. The purpose of this study was to compare the views of community pharmacists on over-the-counter (OTC) omeprazole and simvastatin. METHODS: A pre-piloted questionnaire mailed to 2000 randomly selected community pharmacy premises, with open questions relating to pharmacists' views. Pharmacists' attitudes on aspects of supply were sought using Likert scales. Content analysis was performed on responses to the open question and Wilcoxon signed ranks test (z) used to compare responses to the attitudinal statements. RESULTS: One thousand one hundred and fifty-six questionnaires were returned (57.8%). Themes around omeprazole illustrated positive views compared to concerns relating to simvastatin. Whereas omeprazole was seen as a welcome addition, there were issues around the poor evidence base for simvastatin. Further simvastatin related concerns were in areas of cardiovascular risk assessment, adverse drug reactions and likelihood of patients not committing to therapy. For both agents, excessive cost was an issue. Nearly three-quarters of pharmacists agreed they were entirely confident about selling omeprazole (835, 73%), significantly more than simvastatin (691, 60%); (z = 9.243, p < 0.001). CONCLUSIONS: Two re classified medicines have raised common and product-specific themes. Many themes relate to clinical governance such as evidence-based practice, risk management and continuing professional development (CPD). Views on the availability and circumstances surrounding the OTC supply differ in certain regards, for example, more believe that they should be involved in the sale of simvastatin personally than for sales of omeprazole. PMID- 17918163 TI - Clinical effectiveness of garlic (Allium sativum). AB - The objective of this review is to update and assess the clinical evidence based on rigorous trials of the effectiveness of garlic (A. sativum). Systematic searches were carried out in Medline, Embase, Amed, the Cochrane Database of Systematic Reviews, Natural Standard, and the Natural Medicines Comprehensive Database (search date December 2006). Our own files, the bibliographies of relevant papers and the contents pages of all issues of the review journal FACT were searched for further studies. No language restrictions were imposed. To be included, trials were required to state that they were randomized and double blind. Systematic reviews and meta-analyses of garlic were included if based on the results of randomized, double-blind trials. The literature searches identified six relevant systematic reviews and meta-analysis and double-blind randomized trials (RCT) that were published subsequently. These relate to cancer, common cold, hypercholesterolemia, hypertension, peripheral arterial disease and pre-eclampsia. The evidence based on rigorous clinical trials of garlic is not convincing. For hypercholesterolemia, the reported effects are small and may therefore not be of clinical relevance. For reducing blood pressure, few studies are available and the reported effects are too small to be clinically meaningful. For all other conditions not enough data are available for clinical recommendations. PMID- 17918164 TI - Does garlic have a role as an antidiabetic agent? AB - Diabetes affects a large segment of the population worldwide, and the prevalence of this disease is rapidly increasing. Despite the availability of medication for diabetes, traditional remedies are desirable and are currently being investigated. Garlic (Allium sativum), which is a common cooking spice and has a long history as a folk remedy, has been reported to have antidiabetic activity. However, there is no general agreement on the use of garlic for antidiabetic purposes, primarily because of a lack of scientific evidence from human studies and inconsistent data from animal studies. The validity of data from previous studies of the hypoglycemic effect of garlic in diabetic animals and the preventive effects of garlic on diabetes complications are discussed in this review. The role of garlic as both an insulin secretagogue and as an insulin sensitizer is reviewed. Evidence suggests that garlic's antioxidative, antiinflammatory, and antiglycative properties are responsible for garlic's role in preventing diabetes progression and the development of diabetes-related complications. Large-scale clinical studies with diabetic patients are warranted to confirm the usefulness of garlic in the treatment and prevention of diabetes. PMID- 17918165 TI - Fumonisin B1 alters cell cycle progression and interleukin-2 synthesis in swine peripheral blood mononuclear cells. AB - Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium verticillioides, a fungus that commonly contaminates maize. In the present study, we investigated the effects of FB1 on swine peripheral blood mononuclear cells (PBMC) by measuring cell proliferation, cell cycle progression and interleukin (IL)-2 production. Forty-eight hours after treatment in vitro, FB1 induced a decrease of PBMC proliferation as measured by cell counting and dehydrogenase enzyme activity. This effect was observed starting with 10 microM FB1. The effect of FB1 on cell cycle progression was analyzed by flow cytometry. Incubation of PBMC with FB1 increased the percentage of cells blocked in G0/G1 phase of the cell cycle. Treatment with 200 microM FB1 induces a high blockade of the cell cycle, with 92.4% of cells in G0/G1 phase. This blockade was observed in all lymphocyte subsets tested (CD2+, CD4+, CD8+ and Ig+) as evidenced by dual staining for DNA and membrane surface molecules. A significant decrease of IL2 production was also observed in the supernatants of ConA stimulated PBMC treated with 100 or 200 microM FB1. In conclusion, these data suggest that FB1 may affect immune functions by inhibiting lymphocyte proliferation and IL2 production, suggesting a possible role of FB1 exposure during infectious disease and cancer. PMID- 17918166 TI - Preventive effects of diallyl sulfide on 7,12-dimethylbenz[a]anthracene induced DNA alkylation damage in mouse skin. AB - Mutations that occur through DNA strand breaks are the prerequisites for the development of tumors, which ultimately leads to various genetic disorders including cancer. A number of naturally occurring compounds including certain dietary constituents play an important role in causation and prevention of a number of genetic diseases. Diallyl sulfide (DAS), a volatile organosulfur compound present in garlic has been shown to possess various pharmacological effects including cancer preventive properties. Now we are reporting the antimutagenic properties of DAS on 7,12- dimethylbenz[a]anthracene (DMBA), a carcinogenic polycyclic aromatic hydrocarbon, induced DNA strand breaks in mouse skin, using an alkaline unwinding assay. DAS (2.5-10 mg/kg body-weight) was applied topically, prior and post to DMBA (5 mg/kg body-weight) at the sampling time of 24, 48, 72 and 96 h. DAS application resulted in a significant (p < 0.001) protection in DMBA-induced DNA strand breaks. The pre-treatment of DAS (10 mg/kg body-weight) showed 68.35% protection and post-treatment showed 59.49% protection, at an intermittent period of 48 h, against DMBA-induced DNA strand breakage. These findings suggest that DAS can effectively check the mutations induced by environmental toxicants. PMID- 17918167 TI - Multigram solution-phase synthesis of three diastereomeric tripeptidic second generation dendrons based on (2S,4S)-, (2S,4R)-, and (2R,4S)-4-aminoprolines. AB - Three diastereomeric second-generation (G2) dendrons were prepared by using (2S,4S)-, (2S,4R)-, and (2R,4S)-4-aminoprolines on the multigram scale with highly optimized and fully reproducible solution-phase methods. The peripheral 4 aminoproline branching units of all the dendrons have the 2S,4S configuration throughout, whereas those units at the focal point have the 2S,4S, 2S,4R, and 2R,4S configurations. These latter configurations led to the dendrons being named (2S,4S)-1, (2S,4R)-1, and (2R,4S)-1, respectively. The 4-aminoproline derivatives used in this study are new, although many closely related compounds exist. Their syntheses were optimized. The dendron assembly involved amide coupling, the efficiency of which was also optimized by employing the following well-known reagents: EDC/HOBt, DCC/HOSu, TBTA/HOBt, TBTU/HOBt, BOP/HOBt, pentafluorophenol, and PyBOP/HOBt. It was found that the use of PyBOP is by far the best for dendrons (2S,4S)-1 and (2R,4S)-1, and pentafluorophenol active ester is best for (2S,4R)-1. Because of their multigram scale, all couplings were done in solution instead of by solid-phase procedures. Purifications were, nevertheless, easy. The optical purities of the key intermediates as well as the three G2 dendrons were analyzed by chiral HPLC analysis. These novel, diastereomeric second-generation dendrons have a rather compact and conformationally highly rigid structure that makes them interesting candidates for applications, for example, in the field of dendronized polymers and in organocatalysis. PMID- 17918168 TI - Beneficial effects of sulfated polysaccharides from Sargassum wightii against mitochondrial alterations induced by Cyclosporine A in rat kidney. AB - Sulfated polysaccharides from marine seaweeds are receiving continuous attention owing to their wide therapeutic applications and are known to inhibit free radical generation. It has been well known that mitochondria are the major sources as well as the target of free radicals. The renal tubules have high density of mitochondria and therefore show structural and functional defects in acute renal failure. Hence, the present study is designed to appraise the mitochondrial status during Cyclosporine A (CsA)-induced nephrotoxicity and the effect of sulfated polysaccharides over it. Sulfated polysaccharides (5 mg/kg body weight, subcutaneously) treatment significantly prevented the CsA-induced (25 mg/kg body weight, orally) mitochondrial damage. CsA-induced mitochondrial oxidative stress in rat kidney was evident from increased reactive oxygen species level, decreased antioxidant defense system, coupled with enhanced lipid peroxidation. Further, the activities of tricarboxylic acid cycle and electron transport chain enzymes were decreased in CsA-induced rats, along with a significant increase in the activities of urinary enzymes, thus indicating renal tubular injury. Ultrastructural changes were also in accord with the above aberrations. The above abnormalities were favorably modulated by sulfated polysaccharides supplementation, thus highlighting the significance of sulfated polysaccharides in preventing the renal mitochondrial dysfunction allied with CsA provoked nephrotoxicity. PMID- 17918170 TI - Is garlic a promising food for benign prostatic hyperplasia and prostate cancer? AB - Prostate cancer is one of the most common male malignancies worldwide, and benign prostatic hyperplasia (BPH) is a common cause of lower urinary tract symptoms in elderly men. Garlic (Allium sativum) has been known to have anti-inflammatory, anti-cancer and antioxidant effects. Owing to these effects, garlic and its preparations have been used for the treatment of prostate cancer and relief of BPH symptoms for decades. It is thought that the mechanism(s) through which garlic may show anti-cancer and anti-inflammatory effects should be investigated further. Several researchers are attempting to demonstrate the useful properties of garlic and its mechanism(s) of action. This review aims to present the current studies related with the effects of garlic in prostate diseases, namely prostate cancer and BPH. PMID- 17918171 TI - An extensive class of solids full of holes large enough to enclose over 200 molecules of H2O. PMID- 17918169 TI - Preventing cell death induced by carbonyl stress, oxidative stress or mitochondrial toxins with vitamin B anti-AGE agents. AB - Carbonyls generated by autoxidation of carbohydrates or lipid peroxidation have been implicated in advanced glycation end product (AGE) formation in tissues adversely affected by diabetes complications. Tissue AGE and associated pathology have been decreased by vitamin B(1)/B(6) in trials involving diabetic animal models. To understand the molecular cytoprotective mechanisms involved, the effects of B(1)/B(6) vitamers against cytotoxicity induced by AGE/advanced lipid end product (ALE) carbonyl precursors (glyoxal/acrolein) have been compared to cytotoxicity induced by oxidative stress (hydroperoxide) or mitochondrial toxins (cyanide/copper). Thiamin was found to be best at preventing cell death induced by carbonyl stress and mitochondrial toxins but not oxidative stress cell death suggesting that thiamin pyrophosphate restored pyruvate and alpha-ketoglutarate dehydrogenases inhibited by mitochondrial toxicity. However, B(6) vitamers were most effective at preventing oxidative stress or lipid peroxidation cytotoxicity suggesting that pyridoxal or pyridoxal phosphate were antioxidants and/or Fe/Cu chelators. A therapeutic vitamin cocktail could provide maximal prevention against carbonyl stress toxicity associated with diabetic complications. PMID- 17918172 TI - Total synthesis, configuration, and biological evaluation of anguinomycin C. PMID- 17918173 TI - A sydnone cycloaddition route to pyrazole boronic esters. PMID- 17918174 TI - "Dissolution" of indium(I) iodide: synthesis and structural characterization of the neutral indium sub-halide cluster complex [In6I8(tmeda)4]. PMID- 17918175 TI - Highly improved electroluminescence from a series of novel Eu(III) complexes with functional single-coordinate phosphine oxide ligands: tuning the intramolecular energy transfer, morphology, and carrier injection ability of the complexes. AB - The functional single-coordinate phosphine oxide ligands (4 diphenylaminophenyl)diphenylphosphine oxide (TAPO), (4-naphthalen-1-yl phenylaminophenyl)diphenylphosphine oxide (NaDAPO), and 9-[4 (diphenylphosphinoyl)phenyl]-9H-carbazole (CPPO), as the direct combinations of hole-transporting moieties, and electron-transporting triphenylphosphine oxide (TPPO) were designed and synthesized (amines or carbazole), together with their Eu(III) complexes [Eu(tapo)(2)(tta)(3)] (1), [Eu(nadapo)(2)(tta)(3)] (2), and [Eu(cppo)(2)(tta)(3)] (3; TTA: 2-thenoyltrifluoroacetonate). The investigation indicated that by taking advantage of the modification inertia of the phosphine oxide ligands, the direct introduction of the hole-transport groups as chromophore made TAPO, NaDAPO, and CPPO obtain the most compact structure and mezzo S(1) and T(1) energy levels, which improved the intramolecular energy transfer in their Eu(III) complexes. The amorphous phase of 1-3 proved the weak intermolecular interaction, which resulted in extraordinarily low self-quenching of the complexes. The excellent double-carrier transport ability of the ligands was studied with Gaussian calculations, and the bipolar structure of TAPO and CPPO was proved. The great improvement of the double-carrier transport ability of 1-3 was shown by cyclic voltammetry. Their HOMO and LUMO energy levels of around 5.3 and 3.0 eV, respectively, are the best results for Eu(III) complexes reported so far. A single-layer organic light-emitting diode of 2 had the impressive brightness of 59 cd m(-2) which, to the best of our knowledge, is the highest reported so far. Both of the four-layer devices based on pure 1 and 2 had a maximum brightness of more than 1000 cd m(-2), turn-on voltages lower than 5 V, maximum external quantum yields of more than 3 % and excellent spectral stability. PMID- 17918176 TI - Niobium- and tantalum-based ethylene polymerisation catalysts bearing methylene- or dimethyleneoxa-bridged calixarene ligands. AB - Treatment of p-tert-butylcalix[6]areneH(6) (H(6)tBu-L) or p-tert butylcalix[8]areneH(8) (H(8)tBu-L(1)) with [MCl(5)] (M=Nb, Ta) in refluxing toluene or dichloromethane affords, after work-up, the complexes [{M(NCMe)Cl(2)}(2)(tBu-L)] (M=Nb (1), Ta (2)) and [(MCl(2))(2)(tBu-L(1)H(2))] (M=Nb (4), Ta (5)), respectively. Complex 1, as well as [{Nb(2)(mu-O)(2)(mu Cl)(tBu-LH)}(2)] (3), is also available from [NbOCl(3)] and H(6)tBu-L. Reaction of [MOCl(3)] (M=Nb, Ta) with Li(3)(tBu-L(2)) in diethyl ether, where H(3)tBu-L(2) is p-tert-butylhexahomotrioxacalix[3]areneH(3), affords, after work-up, the trimeric complexes [{M(tBu-L(2))(mu-O)}(3)] (M=Nb (6), Ta (7)). The behaviour of 1 to 7 (not 3), as well as the known complexes [{(MCl)p-tert butylcalix[4]arene}(2)] (M=Nb (8), Ta (9)) and [(MCl(2))p-tert butylcalix[4]arene(OMe)] (M=Nb (10), Ta (11)), as pro-catalysts for the polymerisation of ethylene has been investigated. In the presence of dimethyl (or diethyl)aluminium chloride, methylaluminoxane or trimethylaluminium, these niobium and tantalum procatalysts are all active (<35 g mmol(-1) h(-1) bar(-1)), for the polymerisation of ethylene affording high-molecular-weight linear polyethylene. The dimethyleneoxa-bridged systems (derived from 6 and 7) are more active (84 and 46 g mmol(-1) h(-1) bar(-1), respectively) than the methylene bridged systems. The molecular structures of 1-6 and 10 (acetonitrile solvate) are reported. PMID- 17918177 TI - Exploring the flap pocket of the antimalarial target plasmepsin II: the "55 % rule" applied to enzymes. PMID- 17918178 TI - Downregulation of IFN-gammaR in association with loss of Fas function is linked to tumor progression. AB - The host immune system functions as an intrinsic surveillance network in the recognition and destruction of tumor cells, and it has been demonstrated that lymphocytes and IFN-gamma are the primary tumor suppressors of the immune system. However, the immune system can concurrently select for tumor variants with reduced immunogenicity and aggressive phenotypes. We report here that tumor escape variants that have survived CTL adoptive immunotherapy exhibited decreased expression levels of both Fas and IFN-gammaR in vitro. Furthermore, examination of spontaneously arising mouse primary mammary carcinoma and lung metastases revealed that both Fas and IFN-gammaR protein levels were dramatically lower in lung metastases than in primary tumors in vivo. Functional disruption of either the Fas- or the IFN-gamma signaling pathway enhanced the colonization efficiency of preexisting metastatic tumor cells, whereas disruption of both Fas and IFN gammaR pathways resulted in synergistic augmentation of the colonization efficiency of the preexisting metastatic tumor cells, as determined by experimental lung metastases assay. Gene expression profiling revealed that altered expression of genes involved in immediate IFN-gammaR signaling, the interferon primary response, apoptosis and tumor colonization is associated with loss of IFN-gammaR function and enhanced metastatic potential. Interestingly, disruption of IFN-gammaR function did not alter tumor cell susceptibility to CTL mediated cytotoxicity, but is linked to enhanced infiltration of endogenous T cells in the tumor microenvironment in vivo. These findings suggest that coordinate downregulation of Fas and IFN-gammaR, 2 key components of cancer immunosurveillance system on tumor cells, leads to a more aggressive metastatic phenotype. PMID- 17918180 TI - Akt promotes cisplatin resistance in human ovarian cancer cells through inhibition of p53 phosphorylation and nuclear function. AB - Resistance to cisplatin-based chemotherapy is a major cause of treatment failure in human ovarian cancer. Wild-type TP53 status is often, but not always, associated with cisplatin sensitivity, suggesting that additional factors may be involved. Overexpression/activation of the phosphatidylinositol-3-kinase/Akt pathway is commonly observed in ovarian cancer, and Akt activation is a determinant of chemoresistance in ovarian cancer cells, an effect that may be due, in part, to its inhibitory actions on p53-dependent apoptosis. To that end, we examined the role and regulation of p53 in chemosensitive ovarian cancer cells, as well as in their chemoresistant counterparts, and investigated if and how Akt influences this pathway. Cisplatin induced apoptosis in chemosensitive, but not chemoresistant cells, and this was inhibited by downregulation of p53. Cisplatin upregulated PUMA in a p53-dependent manner, and the presence of PUMA was necessary, but not sufficient for cisplatin-induced apoptosis. p53 was phosphorylated on numerous N-terminal residues, including Ser15, Ser20, in response to cisplatin in chemosensitive, but not chemoresistant cells. Furthermore, activation of Akt inhibited the cisplatin-induced upregulation of PUMA, and suppressed cisplatin-induced p53 phosphorylation, while inhibition of Akt increased total and phospho-p53 contents and sensitized p53 wild-type, chemoresistant cells to cisplatin-induced apoptosis. Finally, mutation of Ser15 and/or Ser20, but not of Ser37, to alanine significantly attenuated the ability of p53 to facilitate CDDP-induced apoptosis, and this was independent of PUMA expression. These results support the hypothesis that p53 is a determinant of CDDP sensitivity, and suggest that Akt contributes to chemoresistance, in part, by attenuating p53-mediated PUMA upregulation and phosphorylation of p53, which are essential, but independent determinants of sensitivity to CDDP-induced apoptosis. PMID- 17918179 TI - Tumor-selective vessel occlusions by platelets after vascular targeting chemotherapy using paclitaxel encapsulated in cationic liposomes. AB - Paclitaxel encapsulated in cationic liposomes (EndoTAG-1) significantly impairs tumor growth by a significant reduction of functional tumor microcirculation and induction of endothelial cell apoptosis. The aim of the study was to analyze whether platelet activation within the tumor microcirculation contributes to the antivascular effects of vascular targeting chemotherapy using EndoTAG-1. In vitro, FACS analysis revealed a significant activation of platelets upon treatment with EndoTAG-1. In vivo, using A-Mel-3 tumors in Syrian Golden hamsters equipped with dorsal skinfold chamber preparations, the contribution of platelets to the antivascular effects of EndoTAG-1 was evaluated by fluorescence and laser scanning microscopy. Immediately after a single treatment with EndoTAG-1 or cationic liposomes devoid of paclitaxel, an increase of platelet adherence in tumor microvessels was observed. This was accompanied by an acute impairment of the microcirculation within the treated tumors leading to reduced tumor perfusion. After repetitive therapy, an increase of platelet adherence and subsequent tumor microvessel occlusions occurred only after treatment with EndoTAG-1. Comparing to "tumor free" normal tissue controls these microthromboses were tumor selective. Significantly disbalancing the coagulation system within tumors by targeted induction of microthromboses within the tumor microcirculation appears to be an important mechanism of EndoTAG-1 therapy. PMID- 17918181 TI - Oldest known case of metastasizing prostate carcinoma diagnosed in the skeleton of a 2,700-year-old Scythian king from Arzhan (Siberia, Russia). AB - To determine whether a 2,700-year-old tumor can be reliably diagnosed using microscopic and proteomic techniques and whether such prostate carcinomas show the same morphological pattern at the micro-level as modern-day carcinomas, this case was investigated. A 40-50-year-old Scythian king who lived during the Iron Age in the steppe of Southern Siberia (Russia) suffered from macroscopically visible osteoblastic and osteoclastic lesions throughout his entire skeleton. Macro-morphological (macroscopy, endoscopy, radiology) and micro-morphological techniques (histology, scanning-electron microscopy) as well as proteomic techniques (1-D- and 2-D-electrophoresis, Western blot) were applied. The results of the morphological and biochemical investigation proved that this mature male suffered for many years from and probably died of a carcinoma of the prostate. The diagnosis mainly rests on the results of the microscopic examination of the lesions and the positive evidence of PSA, which is an important marker for the diagnosis of prostate cancer. It is remarkable that, in this ancient case, the morphological pattern at the microlevel is the same as in recent cases. The loss of the spongy bone substance (red bone marrow) provoked chronic anemia during the final months of the life of this king. The proteomic techniques applied are new for the investigation of recent and ancient macerated bones. Sensitive and reliable biochemical markers (PSA) are an important precondition to detect such tumors in recent and ancient materials. Currently, this is the oldest known case of prostate cancer diagnosed reliably by morphological and biochemical techniques. PMID- 17918182 TI - The use of aromatherapy to treat behavioural problems in dementia. AB - OBJECTIVE: Behavioural and psychological symptoms in dementia (BPSD) are common and distressing to both patients and carers. The use of antipsychotics to treat BPSD is associated with a high burden of side-effects and alternative strategies are required. Aromatherapy is an option that has been recommended for use in dementia. We aimed to review the evidence supporting the use of aromatherapy in BPSD. METHODS: We searched Medline, Cochrane and EMBASE for randomised controlled trials of aromatherapy in patients with dementia. RESULTS: Eleven prospective randomised studies of aromatherapy in BPSD were identified. The aromatherapy oils tested, method of administration and outcome measures used varied widely across the studies. Most of the studies included very small numbers of patients and were designed in such a way that made interpretation of the findings difficult. CONCLUSION: Data supporting the efficacy of aromatherapy are scarce; available studies reported positive and negative consequences for both people with dementia and their carers. The side-effect profile of commonly used oils is virtually unexplored. Although a potentially useful treatment for BPSD, the expectations of clinicians and patients with respect to the efficacy and tolerability of conventional medicines should equally apply to aromatherapy. PMID- 17918183 TI - Outcomes of community-based social service interventions in homebound elders. AB - OBJECTIVE: To document within-client change in function and quality of life over 6 months, and determine whether social service interventions, comorbidity, depressive symptoms, social support and stress are predictors of within-patient change. METHOD: Assess homebound elderly referred for social services on depressive symptoms measured by the Geriatric Depression Scale (GDS), comorbidity with the Charlson Index, and stress and support with the Duke instrument. Function was measured by the Functional Autonomy Scale (FAS), measuring Activities of Daily Living (ADL), Independent Activities of Daily Living (IADL), mobility, communication and mental function. The SF-36 measured quality of life. RESULTS: Among 56 new homebound clients with an average age of 82, 33% had depressive symptoms at baseline (>7 on the GDS). At baseline clients were at or below 25th percentile for five of eight domains of the SF-36, and mental and physical summary scores. Further at baseline, 90% had difficulties with mobility and IADLs; 61% had ADL limitations. At 6-month follow-up overall, 26% had depressive symptoms at follow-up. Greater comorbidity was associated with more depressive symptoms at both baseline and follow-up. By 6 months, 18% had deteriorated on the FAS, while 11% improved. More clients had changes in quality of life; regarding the physical component score, 13% had important deterioration, while 63% improved. Similarly, 33% declined on the mental component while 27% improved. CONCLUSION: Among newly homebound elders, those with significant depressive symptoms are more likely to experience deterioration in function and quality of life over 6 months. However, those with more support showed significant improvement in the SF-36 mental component scale at 6 months. PMID- 17918184 TI - EGFR induces expression of IRF-1 via STAT1 and STAT3 activation leading to growth arrest of human cancer cells. AB - Recently, we reported that epidermal growth factor receptor (EGFR) induce expression of a module of genes known to be inducible by interferons and particularly interferon-gamma. Here we show that the module is tightly regulated by EGFR in the 2 human cancer cell lines that overexpress EGFR, A431 and HN5. The module of genes included the tumor suppressor IRF-1, which was used as a prototypical member to further investigate the regulation and function of the module. Ligand-activated EGFR induce expression of IRF-1 via phosphorylation of STAT1 and STAT3. In contrast, cells expressing the constitutively active cancer specific receptor EGFRvIII are unable to mediate phosphorylation of these STATs and thereby incapable of inducing IRF-1. We also demonstrate that IRF-1 is expressed in an EGF dose-dependent manner, which correlates with inhibition of cell proliferation, and that the regulation of IRF-1 is partially dependent on intracellular Src family kinase activity. Treatment with the dual specific Abl/c Src kinase inhibitor AZD0530 significantly reduces the growth inhibitory effect of high EGF concentrations, signifying that EGFR induced IRF-1 is responsible for the observed growth inhibition. In addition, we show that media from these EGF treated cancer cells upregulate the activation marker CD69 on both B-cells and T cells in peripheral blood. Taken together, these results suggest that cells acquiring sustained high activity of oncogenes such as EGFR are able to activate genes, whose products mediate growth arrest and activate immune effector cells, and which potentially could be involved in alerting the immune system in vivo leading to elimination of the transformed cells. PMID- 17918187 TI - Effect of concomitant treatment with a CYP3A4 inhibitor and a calcium channel blocker. AB - PURPOSE: Grapefruit juice has been found to interact with calcium channel blockers (CCB). This interaction is due to certain nutrients found in grapefruit juice that block the activity of cytochrome P-450 (CYP) 3A4 in the small intestine and liver. Inhibition of CYP3A4 markedly increases bioavailability and increases the risk of an adverse drug reaction (ADR). Many drugs are known to have CYP3A4-blocking activity. This study was performed to investigate whether the concomitant use of a CYP3A4 inhibitor and a CCB in hypertensive patients results in an elevated incidence of ADRs. METHODS: The study included data on 17,430 patients receiving a CCB. Data were obtained from an anti-hypertensive drug database developed by the RAD-AR Council, Japan. A nested case-control design was employed for this study. Cases are defined as patients experiencing an ADR during the follow-up period of 12 weeks. Four controls per case, matched for CCB use, were selected via incidence density sampling. An estimate of the association between the CYP3A4 inhibitor and the ADR was obtained via multivariate conditional logistic regression. RESULTS: Univariate analysis revealed that the odds ratio for experiencing an ADR for the group treated concomitantly with a CCB and a CYP3A4 inhibitor was 1.35 (95% confidence intervals (95%CI), 1.02-1.78), compared with CCB monotherapy. The odds ratio based on multivariate analysis using the 1:4 matched dataset was 1.53 (95%CI, 0.95-2.47) after adjusting for possible confounding factors. CONCLUSIONS: The concomitant treatment with a CYP3A4 inhibitor and a CCB increases the risk of an ADR by 53%, compared with CCB monotherapy. PMID- 17918185 TI - Use of synthetic signal sequences to explore the protein export machinery. AB - The information for correct localization of newly synthesized proteins in both prokaryotes and eukaryotes resides in self-contained, often transportable targeting sequences. Of these, signal sequences specify that a protein should be secreted from a cell or incorporated into the cytoplasmic membrane. A central puzzle is presented by the lack of primary structural homology among signal sequences, although they share common features in their sequences. Synthetic signal peptides have enabled a wide range of studies of how these "zipcodes" for protein secretion are decoded and used to target proteins to the protein machinery that facilitates their translocation across and integration into membranes. We review research on how the information in signal sequences enables their passenger proteins to be correctly and efficiently localized. Synthetic signal peptides have made possible binding and crosslinking studies to explore how selectivity is achieved in recognition by the signal sequence-binding receptors, signal recognition particle, or SRP, which functions in all organisms, and SecA, which functions in prokaryotes and some organelles of prokaryotic origins. While progress has been made, the absence of atomic resolution structures for complexes of signal peptides and their receptors has definitely left many questions to be answered in the future. PMID- 17918186 TI - Alternate PNA-DNA chimeras (PNA-DNA)(n): synthesis, binding properties and biological activity. AB - Peptide nucleic acids (PNAs) are oligonucleotide mimics in which the sugar phosphate backbone has been replaced by a pseudo-peptide backbone. Among PNA based molecules, PNA-DNA conjugates characterized by tracts of DNA bound to N and/or C terminus of PNA are very soluble in aqueous media, are able to recognize exclusively single strands of DNA and RNA in antiparallel fashion, activate RNAse H, bind to transcription factors and are more stable than DNA to nucleases degradation. Very little information is available on chimeras constituted of alternating monomers of PNA and DNA. In this article, we describe a simple fully automated strategy for the synthesis of 6-mer and 10-mer alternate PNA-DNA chimeras consisting of polythymine oligomers, stability assays in fetal calf serum, UV and CD studies of the single strand alternate chimeras and of alternate chimera/DNA and alternate chimera/RNA duplexes. Evidences supporting the formation of duplex hybrids were found. Furthermore, the ability of forming Hoogsteen base pairing with duplex DNA was investigated. Finally, we tested the ability of the PNA-DNA alternates in (a) interfering with reverse transcription of eukaryotic mRNA and (b) inhibiting DNA-protein interactions. PMID- 17918188 TI - Incidents of bee poisoning with pesticides in the United Kingdom, 1994-2003. AB - For over 20 years, the UK Agriculture Departments have monitored the direct effects of pesticides on beneficial insects, mainly honeybees (Apis mellifera, L.) and bumblebees (Bombus terrestris, L.), as part of the Wildlife Incident Investigation Scheme (WIIS). The Central Science Laboratory (CSL) has contributed to WIIS by providing the required laboratory skills for the determination of bee diseases and the expert analytical experience necessary to determine low-level pesticide residues and interpret these results. The results from WIIS form part of the pesticide regulatory process coordinated by the Pesticides Safety Directorate (PSD) and are published each year. This paper has reviewed the data from WIIS over the 10 year period from 1994 to 2003. The overall trend is that suspected poisoning incidents, reported by beekeepers and the general public, have declined from 56 incidents per year to 23 incidents per year. The number of these incidents that have been attributed to pesticide poisoning has also declined, from 25 incidents to five incidents per year. The possible reasons for these changes and the circumstances involved in the bee poisoning incidents are discussed. However, the source of the pesticide in bee poisoning incidents is often uncertain and the likely cause of these incidents and any trends over time are also discussed. PMID- 17918189 TI - In focus: pesticides and honeybees -- the work of the ICP-BR Bee Protection Group. PMID- 17918190 TI - Hazards of pesticides to bees. Proceedings of the 9th International Symposium of the International Commission for Plant-Bee Relationships Bee Protection Group. October 12-14, 2005. Sand Hutton, York, United Kingdom. PMID- 17918191 TI - Pulsed electrical stimulation for control of vasculature: temporary vasoconstriction and permanent thrombosis. AB - A variety of medical procedures is aimed to selectively compromise or destroy vascular function. Such procedures include cancer therapies, treatments of cutaneous vascular disorders, and temporary hemostasis during surgery. Currently, technologies such as lasers, cryosurgery and radio frequency coagulation, produce significant collateral damage due to the thermal nature of these interactions and corresponding heat exchange with surrounding tissues. We describe a non-thermal method of inducing temporary vasoconstriction and permanent thrombosis using short pulse (microseconds) electrical stimulation. The current density required for vasoconstriction increases with decreasing pulse duration approximately as t( 0.25). The threshold of electroporation has a steeper dependence on pulse duration-exceeding t(-0.5). At pulse durations shorter than 5 micros, damage threshold exceeds the vasoconstriction threshold, thus allowing for temporary hemostasis without direct damage to surrounding tissue. With a pulse repetition rate of 0.1 Hz, vasoconstriction is achieved approximately 1 min after the beginning of treatment in both arteries and veins. Thrombosis occurs at higher electric fields, and its threshold increases with vessel diameter. Histology demonstrated a lack of tissue damage during vasoconstriction, but vascular endothelium was damaged during thrombosis. The temperature increase does not exceed 0.1 degrees C during these treatments. PMID- 17918193 TI - Neuropsychological testing for the assessment of manganese neurotoxicity: a review and a proposal. AB - BACKGROUND: Manganese exposure can cause neurobehavioral and neurological signs and symptoms. A review of the literature was carried out in order to identify the most sensitive and effective methods to assess these outcomes. METHODS: Studies on manganese exposure performed with neuropsychological tests were reviewed using Medline. Test methods, obtained outcomes, and dose-response relationships were considered. Based on the review and additional considerations, a testing battery for adult and children was identified. RESULTS: A total number of 31 studies were reviewed and divided in 18 occupational, 7 environmental, and 6 on children. A large variety of tests exploring motor and cognitive functions and mood were applied. Neurobehavioral effects were dose-dependent in several studies and determined by much lower exposure levels compared to 1 mg Mn/m(3) in total dust which is considered to be the minimum concentration able to induce the classical clinical features of "manganism." Although through a wide variety of tests, the observed outcomes support the knowledge that manganese exposure can cause neurobehavioral effects. Motor functions are mainly affected in adults, and cognitive functions and behavior are more affected in children. CONCLUSIONS: Literature on manganese neurobehavioral effect is quite consistent, however, further improvement may be achieved by using better structured and more comparable evaluation methods. Based on the analysis of the existing experiences a test battery including tests of motor functions, response speed, cognitive functions, intellectual abilities, mood, and symptom questionnaires is suggested. We recommend that the tests indicated should always be included in future studies as a core battery. PMID- 17918194 TI - Genetic and environmental contributions to stability in loneliness throughout childhood. AB - Heritability estimates based on two small cross-sectional studies in children indicate that the genetic contribution to individual differences in loneliness is approximately 50%. A recent study estimated the genetic contribution to variation in loneliness in adults to be 48%. The current study aims to replicate and expand these findings by conducting longitudinal analyses in order to study causes of individual differences in stability of loneliness throughout childhood. Univariate and multivariate longitudinal analyses are conducted in a large sample of young Dutch twins. Information on loneliness comes from maternal ratings on the Child Behavior Checklist. Using an average score of loneliness over ages 7, 10, and 12, results from the two previous studies are replicated and a heritability estimate of 45% is found. The remaining variance is accounted for by shared environmental influences (12%), and nonshared environmental influences (43%). The longitudinal analyses, however, show that heritability is 58% at age 7, 56% at age 10, but drops to 26% at age 12. A parallel increase in influences of shared family environment is observed, explaining 6% of the variance at age 7, 8% at age 10 and 35% at age 12. The remaining variance is explained by relatively stable influences of nonshared environmental factors. Stability in loneliness is high, with phenotypic correlations in the range of 0.51-0.69. This phenotypic stability is mainly caused by genetic and nonshared environmental influences. The results indicate the importance of both innate as well as nonshared environmental factors for individual differences in loneliness. Further, different results between causes of individual differences for the average score of loneliness and results for age 12 from the longitudinal analyses, indicate the importance of longitudinal analyses with data at well-defined ages. PMID- 17918192 TI - Loss of the eukaryotic initiation factor 3f in pancreatic cancer. AB - Aberrant regulation of the translation initiation is known to contribute to tumorigenesis. eIF3 plays an important role in translation initiation. eIF3f is the p47 subunit of the eIF3 complex whose function in cancer is not clear. Initial studies from our group indicated that eIF3f expression is decreased in pancreatic cancer. Overexpression of eIF3f induces apoptosis in pancreatic cancer cells. The eIF3f gene is located at chromosome band region 11p15.4. Loss of 11p15.4 is a common event in many tumors including pancreatic cancer. In order to investigate the molecular mechanism of the decreased expression of eIF3f in pancreatic cancer, we performed loss of heterozygosity (LOH) analysis in 32 pancreatic cancer specimens using three microsatellite markers encompassing the eIF3f gene. We showed that the prevalence of LOH ranged from 71% to 93%. We also performed eIF3f gene copy number analysis using quantitative real time PCR to further confirm the specific allelic loss of eIF3f gene in pancreatic cancer. We demonstrated a statistically significant decrease of eIF3f gene copy number in pancreatic tumors compared with normal tissues with a tumor/normal ratio of 0.24. Furthermore, RNA in situ hybridization and tissue microarray immunohistochemistry analysis demonstrated that eIF3f expression is significantly decreased in human pancreatic adenocarcinoma tissues compared to normal pancreatic tissues. These data provides new insight into the understanding of the molecular pathogenesis of eIF3f during pancreatic tumorigenesis. PMID- 17918195 TI - Repeated confidence intervals for adaptive group sequential trials. AB - This paper proposes a method for computing conservative confidence intervals for a group sequential test in which an adaptive design change is made one or more times over the course of the trial. The key idea, due to Muller and Schafer (Biometrics 2001; 57:886-891), is that by preserving the null conditional rejection probability of the remainder of the trial at the time of each adaptive change, the overall type I error rate, taken unconditionally over all possible design modifications, is also preserved. We show how this principle may be extended to construct one-sided confidence intervals by applying the idea to a sequence of dual tests derived from the repeated confidence intervals (RCIs) proposed by Jennison and Turnbull (J. Roy. Statist. Soc. B 1989; 51:301-361). These adaptive RCIs, such as their classical counterparts, have the advantage that they preserve the desired coverage probability even if the pre-specified stopping rule is over-ruled. The statistical methodology is explored by simulations and is illustrated by an application to a clinical trial of deep brain stimulation for Parkinson's disease. PMID- 17918199 TI - Deletion of the LIME adaptor protein minimally affects T and B cell development and function. AB - LIME (Lck-interacting membrane protein) is a transmembrane adaptor that associates with the Lck and Fyn protein tyrosine kinases and with the C-terminal Src kinase (Csk). To delineate the role of LIME in vivo, LIME-deficient mice were generated. Although Lime transcripts were expressed in immature and mature B and T cells, the absence of LIME impeded neither the development nor the function of B and T cells. TCR transgenic mice deprived of LIME showed, however, a 1.8-fold enhancement in positive selection. Since B cells and activated T cells express LIME and the related adaptor NTAL, mice lacking both adaptors were generated. Double-deficient mice showed no defect in the development and function of B and T cells, and the lack of LIME had no effect on the autoimmune syndrome that develops in aged NTAL-deficient mice. In contrast to a previous report, we further showed that this autoimmune syndrome develops in the absence of T cells. Therefore, our in vivo results refute all the previous roles postulated for LIME on the basis of studies of transformed B and T cells and demonstrate that LIME has no seminal role in the signaling cassette operated by antigen receptors and coreceptors. PMID- 17918200 TI - IL-21 regulates experimental colitis by modulating the balance between Treg and Th17 cells. AB - Regulatory T (T(reg)) cells play a key role in the maintenance of the immune system homeostasis. T(reg) cells can be generated in the periphery under control of TGF-beta, a cytokine involved in the negative control of the immune system. However, TGF-beta cooperates with IL-6 in the generation of Th17 cells, a novel class of effector cells involved in numerous inflammatory diseases, including colitis. Therefore, TGF-beta emerges as a mediator of both anti-inflammatory and pro-inflammatory processes, depending on the local cytokine milieu. Here we demonstrate that IL-21, a type-1 cytokine produced by T cells and involved in the pathogenesis of immune-mediated diseases, prevents the TGF-beta-dependent expression of FoxP3, the master regulator of T(reg) cell commitment, and the induction of suppressive capacity in naive CD4(+) T cells, while promoting the differentiation of Th17 cells. In vivo, CD4(+) naive T cells activated in the presence of TGF-beta and IL-21 failed to suppress colitis while inducing an inflammatory response characterized by high levels of IL-17 and RORgammat, the transcription factor expressed by Th17 cells. Therefore, IL-21 emerges as a key modulator of TGF-beta signaling, leading to the reciprocal differentiation of T(reg) and Th17 cells. PMID- 17918202 TI - Overexpression of IL-21 promotes massive CD8+ memory T cell accumulation. AB - The ability of IL-21 to promote in vitro T cell survival led us to investigate its biological activity in vivo. We report that overexpression of IL-21 in transgenic mice drives CD8(+) memory T cell accumulation with a concomitant reduction in naive T cell numbers. These memory T cells are functional, given their ability to rapidly produce IFN-gamma and proliferate following stimulation. Since the homeostasis of naive and memory T cells is controlled by cytokines, we evaluated whether IL-21 influences cytokine receptor expression. We show that IL 21 inhibits IL-7R expression on naive T cells in vitro, suggesting impaired IL-7 mediated naive T cell survival in IL-21-transgenic mice. In contrast, IL-7R expression on CD4(+) memory T cells is not affected, allowing their IL-7 dependent survival in IL-21-transgenic mice. Although IL-21 decreases IL-7R expression on CD8(+) memory T cells, this has no impact on their survival since their maintenance in the T cell pool is IL-7-independent. Rather, we demonstrate that CD8(+) memory T cells are receptive to IL-21 survival signals allowing for their accumulation in IL-21-transgenic mice. This study identifies new roles for IL-21 in T cell homeostasis and in the regulation of T cell responses to cytokines. PMID- 17918201 TI - TLR-mediated stimulation of APC: Distinct cytokine responses of B cells and dendritic cells. AB - In addition to their role in humoral immunity, B lymphocytes are important antigen-presenting cells (APC). In the same way as other APC, B cells make cytokines upon activation and have the potential to modulate T cell responses. In this study, we investigated which mouse B cell subsets are the most potent cytokine producers, and examined the role of Toll-like receptors (TLR) in the control of secretion of IL-6, IL-10, IL-12 and IFN-gamma by B cells. Production of some cytokines was restricted to particular subsets. Marginal zone and B1 cells were the predominant source of B cell IL-10 in the spleen. Conversely, follicular B cells were found to express IFN-gamma mRNA directly ex vivo. The nature of the activating stimulus dramatically influenced the cytokine made by B cells. Thus, in response to combined TLR stimulation, or via phorbol esters, IFN gamma was secreted. IL-10 was elicited by T-dependent activation or stimulation through TLR2, 4 or 9. This pattern of cytokine expression contrasts with that elicited from dendritic cells. QRT-PCR array data indicate that this may be due to differential expression of TLR signalling molecules, effectors and adaptors. Our data highlight the potentially unique nature of immune modulation when B cells act as APC. PMID- 17918204 TI - A critical lineage-nonspecific role for pTalpha in mediating allelic and isotypic exclusion in TCRbeta-transgenic mice. AB - Although it is well established that early expression of TCRbeta transgenes in the thymus leads to efficient inhibition of both endogenous TCRbeta and TCRgamma rearrangement (also known as allelic and "isotypic" exclusion, respectively) the role of pTalpha in these processes remains controversial. Here, we have systematically re-evaluated this issue using three independent strains of TCRbeta transgenic mice that differ widely in transgene expression levels, and a sensitive intracellular staining assay that detects endogenous TCRVbeta expression in individual immature thymocytes. In the absence of pTalpha, both allelic and isotypic exclusion were reversed in all three TCRbeta-transgenic strains, clearly demonstrating a general requirement for pre-TCR signaling in the inhibition of endogenous TCRbeta and TCRgamma rearrangement. Both allelic and isotypic exclusion were pTalpha dose dependent when transgenic TCRbeta levels were subphysiological. Moreover, pTalpha-dependent allelic and isotypic exclusion occurred in both alphabeta and gammadelta T cell lineages, indicating that pre TCR signaling can potentially be functional in gammadelta precursors. Finally, levels of endogenous RAG1 and RAG2 were not down-regulated in TCRbeta-transgenic immature thymocytes undergoing allelic or isotypic exclusion. Collectively, our data reveal a critical but lineage-nonspecific role for pTalpha in mediating both allelic and isotypic exclusion in TCRbeta-transgenic mice. PMID- 17918203 TI - Relevant antibody subsets against MOG recognize conformational epitopes exclusively exposed in solid-phase ELISA. AB - A pathogenic role for circulating anti-myelin antibodies is difficult to establish in multiple sclerosis (MS). Here, we unravel a broad heterogeneity within the anti-myelin oligodendrocyte glycoprotein (MOG) antibodies in humans and non-human primates, and demonstrate that detection of important epitopes of MOG within the pathogenic repertoire is exclusively dependent on presentation on a solid-phase MOG conformer. Results of ELISA and those of a liquid-phase assay were compared using a MOG protein with identical sequence but different conformations. We tested sera from 50 human subjects, plasma of Callithrix jacchus marmosets known to contain antibodies reactive to either conformational or linearized MOG, and monoclonal, conformation-dependent anti-MOG antibodies. We have found no antibody reactivity against the soluble MOG conformer in human serum, and show that this lack of detection is not due to technical artifacts. Rather, dominant epitopes of MOG are not displayed in soluble phase, as shown by a complete lack of binding of conformation-dependent mAb. In MP4-immune marmosets that exhibit demyelinating pathology due to spreading of antibody determinants to myelin-embedded MOG, only ELISA can detect pathogenic circulating anti-MOG antibodies. Thus, the accurate detection of important subsets of pathogenic anti MOG antibodies requires methods in which MOG is displayed similarly to its natural conformation in myelin. PMID- 17918205 TI - Both CD133+ and CD133- medulloblastoma cell lines express ligands for triggering NK receptors and are susceptible to NK-mediated cytotoxicity. AB - Adoptive cellular immunotherapy has been proposed as an additional treatment of medulloblastoma, an intracranial tumor characterized by a particularly poor prognosis. However, little is known on the ability of the immune system to effectively attack this tumor. In this study, we show that activated human NK cells efficiently kill medulloblastoma cell lines in vitro. NK-mediated killing involved different activating receptors (including NKp46, NKp30, DNAM-1 and NKG2D) and correlated with the presence of their specific ligands on tumor cells. In contrast, the absence of major adhesion interactions, such as LFA-1/ICAM did not impair the NK-mediated cytotoxicity. Medulloblastoma expressed a number of tumor-associated molecules including CD146 and CD133, considered a marker for cancer stem cells. Remarkably, both CD133-positive and CD133-negative cell lines were susceptible to lysis. Tumor cells also expressed molecules that are currently used as diagnostic tools for neuroblastoma cell identification. In particular, B7 homolog 3 (B7-H3) was expressed by all the medulloblastoma cell lines analyzed, while the presence of GD(2) and NB84 was restricted to given cell lines and/or marked a defined tumor cell subset. PMID- 17918207 TI - p53 Codon 72 polymorphism and the risk of esophageal squamous cell carcinoma. AB - The polymorphisms of the tumor suppressor gene p53 have been extensively investigated in numerous malignant tumors, particularly carcinomas associated with human papillomavirus (HPV) infection. However, the results remain controversial. To address a potential correlation between the p53 genotypes and the risk of esophageal squamous cell carcinoma (ESCC), we investigated the p53 codon 72 polymorphism in 435 patients with ESCC and 550 cancer-free subjects from the same geographical region. p53 Arg/Arg genotype was significantly increased in ESCC cases compared with control subjects (85.7 vs. 49.6%, P < 0.001), resulting in an elevated ESCC risk (OR = 6.48, 95% CI = 4.65-9.03). In addition, among p53 Arg/Arg carriers, HPV infection, smoking, and drinking might further increase the risk of ESCC development. PMID- 17918206 TI - Polarization transfer for sensitivity-enhanced MRS using a single radio frequency transmit channel. AB - Polarization transfer techniques are used to enhance sensitivity and improve localization in multinuclear MRS, by transferring polarization from highly polarized or even hyperpolarized nuclei to less sensitive spin systems. Clinical MR scanners are in general not equipped with a second radio frequency (RF) transmit channel, making the conventional implementation of polarization transfer techniques such as distortionless enhanced polarization transfer (DEPT) impossible. Here we present a DEPT sequence using pulses sequentially that can be used on a single RF transmit channel (SC-DEPT). Theoretical simulations, phantom measurements, and in vivo results from human brain at 3 T show that the SC-DEPT method performs as well as the conventional DEPT method. The results indicate that an independent second RF transmit channel for simultaneous pulsing at different nuclear frequencies is not needed for polarization transfer, facilitating the use of these methods with common clinical systems with minor modifications in the RF architecture. PMID- 17918208 TI - Retinol decreases phosphatidylinositol 3-kinase activity in colon cancer cells. AB - Previously, we showed that retinol inhibited all-trans-retinoic acid (ATRA) resistant human colon cancer cell invasion via a retinoic acid receptor independent mechanism. Because phosphatidylinositol 3-kinase (PI3K) regulates cell invasion, the objective of the current study was to determine if retinol affected PI3K activity. Following 24 h of serum starvation, the ATRA resistant human colon cancer cell lines HCT-116 and SW620 were treated with 0, 1, or 10 microM retinol. Thirty minutes of retinol treatment resulted in a significant decrease in PI3K activity in both cell lines. To determine the mechanism by which retinol reduces PI3K activity, the levels and heterodimerization of the regulatory subunit, p85, and the catalytic subunit, p110, of PI3K were examined. Retinol treatment did not alter p85 or p110 protein levels or the heterodimerization of these subunits at any time point examined. To determine if retinol affected the ability of PI3K to phosphorylate the substrate, phosphatidylinositol (PI), PI3K was immunoprecipitated from control cells and incubated with 10 microg PI and increasing concentrations of retinol or 10 microg retinol and increasing concentrations of PI. Retinol decreased PI3K activity in a dose-responsive manner and increased PI suppressed the inhibitory effect of retinol on PI3K activity. Finally, the PI3K inhibitor, LY294002, mimicked the ability of retinol to decrease cell invasion. Computational modeling revealed that retinol may inhibit PI3K activity in a manner similar to that of wortmannin. Thus, a decrease in PI3K activity due to retinol treatment may confer the ability of retinol to inhibit ATRA-resistant colon cancer cell invasion. PMID- 17918209 TI - Expression of nuclear Notch3 in pancreatic adenocarcinomas is associated with adverse clinical features, and correlates with the expression of STAT3 and phosphorylated Akt. AB - BACKGROUND AND OBJECTIVES: Reactivation of the Notch signalling pathway occurs in a range of human malignancies. Previous research suggests that Notch3 is expressed in pancreatic adenocarcinomas, but neither cellular location nor association with clinical parameters has been described. The relationship between Notch3, clinical endpoints, and other proteins with potential to interact with Notch was therefore examined. METHODS: An immunohistochemical study was performed on human pancreatic adenocarcinoma (n = 23) and normal pancreas (n = 12), to assess expression of Notch3, cyclin D1, pAkt, STAT3 and pSTAT3. Immunohistochemical data were then correlated with clinicopathological characteristics. RESULTS: Notch3 was significantly overexpressed in the cytoplasm of 73.9% of tumours. Nuclear expression was not observed in normal pancreatic ductal tissue, but was noted in 43.5% of tumours. No tumour expressing nuclear Notch3 was resectable. There were significant correlations between expression and intracellular location of Notch3 and each of STAT3, pSTAT3 and pAkt, but not cyclin D1. CONCLUSION: The presence of Notch3 in tumour nuclei is likely to represent functional activation of the protein, and is clearly linked to a more aggressive tumour phenotype. The correlation with STAT3, pSTAT3 and pAkt expression has not previously been described and the concurrent intracellular localisation of these proteins suggests a functional relationship between them. PMID- 17918210 TI - Mass fingerprinting of toxic fractions from the venom of the Indian red scorpion, Mesobuthus tamulus: biotope-specific variation in the expression of venom peptides. AB - The red scorpion, Mesobuthus tamulus, is found in two distinct biotopes within the Indian state of Maharastra-a tropical, sea-level biotope and a semi-arid biotope, up to 600 m. Scorpions from these two geographical areas show marked differences in toxicity. Using mass spectrometry, we have shown biotope-specific variation in the expression of peptides from scorpions collected from these two distinct areas. Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS) and reversed-phase liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) were assessed as techniques for obtaining mass fingerprint data. On line LC/ESI-MS was judged to be the method of choice and unique biotope-specific mass fingerprints, with key diagnostic markers, were obtained. PMID- 17918211 TI - From lead to manganese through mercury: mythology, science, and lessons for prevention. AB - BACKGROUND: Lead (Pb), mercury (Hg), and manganese (Mn) are well-known neurotoxic metals. The knowledge of toxicity was developed through an extensive amount of research, starting with lead and mercury and proceeding today with manganese. Unfortunately, the consequent implementation of preventive measures was generally delayed, causing important negative effects to the exposed populations. METHODS: A review and historical reconstruction of the research development that yielded modern understanding of lead and mercury neurotoxicity was conducted to derive useful lessons for the prevention of manganese neurotoxicity. RESULTS: Medieval alchemists named planets and metals from gods since they were already aware of the toxicity and the adverse effects caused by lead and mercury. Historical lessons learned from these two metals may help to avoid the repetition of further mistakes regarding other neurotoxic metals like manganese. The knowledge and experience on the toxicokinetics and toxicodynamics of lead and mercury is useful and valuable to identify a proper approach to "safe" exposure levels for manganese. CONCLUSIONS: Further information is still needed on the early neurotoxic and neurobehavioral effects after prolonged exposure to very low doses of lead, mercury, and manganese. Nevertheless, according to the precautionary principle, effective preventive measures should be already undertaken to prevent the onset of more severe health effects in the population. This is the most important lesson to be learned and applied from more than 30 years of occupational and environmental neurotoxicology of metals. PMID- 17918212 TI - Behavioral effects of subchronic inorganic manganese exposure in rats. AB - BACKGROUND: Manganese, an essential micronutrient, is a potential neurotoxicant in prolonged overexposure. Parkinson-like syndrome, motor deficit, disturbed psychomotor development are typical signs of neuropathological alterations due to Mn in humans. METHODS: Young adult rats, in three groups of 16 each, received 15 and 59 mg/kg b.w. MnCl(2), (control: distilled water) via gavage for 10 weeks, and were kept for further 12 weeks. Correlation of MnCl(2) exposure to body and organ weights, neurobehavioral effects (spatial memory, exploratory activity, psychomotor performance, pre-pulse inhibition), and histopathological changes (gliosis) was sought. RESULTS: By the end of treatment, Mn accumulated in blood, cortex, hippocampus, and parenchymal tissues. Body and organ weights were reduced in high dose rats. All treated rats showed hypoactivity, decreased memory performance, and diminished sensorimotor reaction. In the dentate gyrus of these, GFAP immunoreactivity increased. During the post-treatment period, body weight of the high dose group remained decreased, locomotor activity returned to control, but the lasting effect of MnCl(2) could be revealed by amphetamine. CONCLUSION: Using complex methodology, new data were obtained regarding the relationship between the long-term effects of MnCl(2) at neuronal and behavioral level. PMID- 17918213 TI - Direct visualisation of peptide hormones in cultured pancreatic islet alpha- and beta-cells by intact-cell mass spectrometry. AB - The application of intact-cell mass spectrometry (ICM) by matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry to achieve direct protein-profiling of bacterial species is now well established. However, this methodology has not to our knowledge been applied to the analysis of mammalian cells in routine culture. Here, we describe a novel application of ICM by which we have identified proteins in intact cells from two lines representative of pancreatic islet alpha- and beta-cells. Adherent alphaTC1 clone 9 and betaTC6 F7 cells were harvested into phosphate-buffered saline (PBS) using enzyme-free dissociation buffer before 1 microL of cell suspension was spotted onto MALDI plates. Cells were overlaid with sinapinic acid then washed with pure water before application of a final coat of sinapinic acid. Data in the 2000 20,000 m/z range were acquired in linear mode on a Voyager DE-Pro mass spectrometer. The proteins which ionised were composed in large part of peptide hormones (e.g. insulin and glucagon) known to be packaged into the secretory granules of the beta- and alpha-cells respectively. However, in addition to visualising the peptides expected to be associated with these cells, a mass consistent with oxyntomodulin was identified in the cultured alpha-cells, a finding not previously reported to our knowledge. In summary, this paper describes, for the first time, a rapid and direct method useful for identifying secretory products in intact endocrine cells. PMID- 17918215 TI - High prevalence of Parkinsonian disorders associated to manganese exposure in the vicinities of ferroalloy industries. AB - OBJECTIVE: To assess the prevalence of Parkinsonian disturbances in relationship to environmental exposure to manganese due to ferroalloy industries in the province of Brescia, Northern Italy. METHODS: Manganese concentrations were measured in settled dust collected in each of the 206 municipalities. Parkinsonian patients were identified using two sources: (1) clinical registers from local hospitals, specialized neurologists, and exemption from prescription payment; (2) L-Dopa prescriptions. Standardized prevalence rates and raw and full Bayesian-smoothed standardized morbidity ratios (SMRs) were calculated for the entire province and for each municipality. RESULTS: Manganese concentrations in settled dust were significantly higher in the surroundings and downwind from the industrial plants. A total number of 2,677 Parkinsonian cases were identified among 903,997 residents (crude prevalence, 296/100,000; 95% CI: 284.80-307.20; standardized prevalence, 407/100,000; 95% CI: 393.87-420.12). Significantly higher SMRs (Kruskal-Wallis chi(2) 1 df = 17.55, P < 0.001) were observed in 37 municipalities in the vicinities of ferromanganese plants (324 cases among 77,708 residents; standardized prevalence 492/100,000; 95% CI: 442.80-541.20), compared to the other 169 municipalities of the province (2,353 cases among 826,289 residents, standardized prevalence 321/100,000; 95% CI 308.80-333.20). Row and Bayesian SMRs were associated with the concentrations of manganese in settled dust. CONCLUSION: Study results suggest that environmental exposure to manganese is associated with an increased prevalence of Parkinsonian disturbances. Since the highest prevalence rates were observed in a closed community of the pre-Alps where the industries are located, further research should address a possible interactive role of genetic factors. PMID- 17918214 TI - Identification of protein phosphorylation sites within Ser/Thr-rich cluster domains using site-directed mutagenesis and hybrid linear quadrupole ion trap Fourier transform ion cyclotron resonance mass spectrometry. AB - We describe a method for the analysis of multi-site phosphorylation in serine/threonine (Ser/Thr)-rich protein sequences. Site-specific mutagenesis was used to introduce tryptic cleavage sites in the serine glutamine/threonine glutamine cluster domain (SCD) of the human checkpoint protein kinase (Chk2). The mutant proteins were shown to autophosphorylate on residues that are inducibly phosphorylated when mammalian cells are exposed to ionizing radiation (serine 33/35, serine 516, threonine 68 and threonine 432). Five Ser/Thr clusters within the SCD were flanked by arginine or lysine residues to produce tryptic peptides for nanospray liquid chromatography (nanoLC)/linear quadrupole ion trap Fourier transform ion cyclotron resonance mass spectrometry. Phosphorylation sites were assigned using accurate-mass-driven analysis and interpretation of low-energy collision-induced dissociation spectra acquired in the ion trap. In addition to verifying known phosphorylation sites, seventeen novel sites were identified within the SCD of Chk2. The approach should be applicable to other O-linked post translational modifications that occur in proteins with Ser/Thr-rich sequences. PMID- 17918216 TI - Curing hepatocellular carcinomatosis with biologic therapy and surgery: achieving the impossible? AB - Hepatocellular carcinoma (HCC) that present as hemoperitoneum are uncommon, often recur as peritoneal carcinomatosis, and have poor prognosis. Systemic chemotherapies are minimally effective in advanced HCC and less so as neo adjuvant therapy. This report describes a 47-year woman with ruptured HCC who developed recurrent carcinomatosis. She received biologically targeted systemic therapy which resulted in significant radiographic tumor response. Salvage surgery revealed complete pathologic response in all tumor nodules, thus the patient was rendered cancer-free. PMID- 17918217 TI - Sialolith characterization by scanning electron microscopy and X-ray photoelectron spectroscopy. AB - The objective of this study has been to characterize sialolith, a calcium phosphate deposit that develops in the human oral cavity, by high-resolution field emission scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The morphological and chemical data obtained helped in the determination of their formation mechanism in salivary glands. Sialoliths in the submandibular salivary glands may arise secondary to sialodenitis, but not via a luminal organic nidus. We believe this is the first study that characterizes a sialolith by XPS. PMID- 17918224 TI - The value of expert second opinion in diagnosis of soft tissue sarcomas. AB - INTRODUCTION: Soft tissue sarcomas represent a heterogeneous group of tumours with a wide range of clinical behaviour. Exact determination of diagnosis and prognosis is critical in order to guide surgical decisions and provide systemic therapy or radiation for patients. The value of consultative second opinions has been proven for general surgical pathology; some studies suggest an even higher value for the soft tissue tumour specimens in particular. METHODS: We reviewed 603 patients who were operated on at our institution with the diagnosis of soft tissue sarcoma and aggressive fibromatosis; we focused on mismatches in primary and definite tumour-entity and -grading with respect to the diagnosing institution and the primary surgical procedure. RESULTS: We found concordant primary diagnosis in 28.3% for pathologists in private clinics, 29.6% for hospital pathologists, 36.8% for academic medical centres (university hospitals) and 70.5% for the Department of Pathology at our institution. An improvement in diagnosis or confirmation of the correct primary diagnosis by the second opinion was seen in 73.1% of the patients; in 2.5%, the second opinion was false. DISCUSSION: For accurate determination of prognosis and to provide optimal therapeutic decisions we consider expert second opinion essential for optimal treatment of soft tissue sarcomas. PMID- 17918223 TI - A comparison of the perceptions and beliefs of workers and owners with regard to workplace safety in small metal fabrication businesses. AB - BACKGROUND: Problems of improving safety in small business establishments may include a lack of resources, limited unionization, and an informal management structure. METHODS: We evaluated worker and manager perceptions of worksite health and safety using Social Cognitive Theory. We used a business safety scorecard to audit the safety conditions, policies and programs, and work practices. Comparisons were made between the different measures. RESULTS: Businesses with safety committees had 1.7-2.1 times higher proportion of positive safety scorecard items than businesses without committees. Union status and business size were not associated with business safety audit results. Non-English speaking and less educated employees reported higher levels of knowledge about safety than did their more educated and/or English-speaking peers. CONCLUSIONS: The presence of a safety committee is the single most important indicator of workplace safety. Self-reported understanding of workplace safety is greater among employees who do not speak English or have lower levels of formal education. Future worksite interventions should consider the need for participatory worksite safety committees. Multilingual training programs would help reach a greater proportion of workers. PMID- 17918225 TI - Management of penile cancer. PMID- 17918226 TI - Breast cancer screening compliance among young women in a free access healthcare system. AB - BACKGROUND AND OBJECTIVES: To examine mammographic screening compliance among young military healthcare beneficiaries and to examine factors related to one time and recent mammographic compliance. METHODS: Medical records were reviewed for 1,073 subjects (age 41-47) recording dates of the two most recent screening mammograms. Examined outcomes were: whether the woman ever had mammography and, if so, whether she had a mammogram within 400 days. Examined predictors were: ethnicity, age, Gail Model risk score, family history, whether the woman knew a young woman with breast cancer, and importance attributed to breast cancer screening. RESULTS: 90.4% of women studied had at least one mammogram. 71.1% underwent screening within 400 days. Rates of ever having mammography were higher for women with family history of breast cancer and Asian, Pacific Islander, Black or Hispanic women. No measured covariate correlated with having mammography within 400 days. CONCLUSIONS: One time screening participation was high in this select group of women for whom cost and access barriers were removed, but was lower with regard to having a recent mammogram. Correlates of ever having and recent mammography are not synonymous. PMID- 17918227 TI - Mortality among workers at the Savannah River Site. AB - BACKGROUND: Workers employed at the Savannah River Site (SRS) were potentially exposed to a range of chemical and physical hazards, many of which are poorly characterized. We therefore compared the observed deaths among workers to expectations based upon death rates for referent populations. METHODS: The cohort included 18,883 SRS workers hired between 1950 and 1986. Vital status and cause of death information were ascertained through 2002. Sex-specific standardized mortality ratios (SMR) were computed using U.S. and South Carolina mortality rates. SMRs were tabulated separately for monthly-, weekly-, and hourly-paid men. RESULTS: Males had fewer deaths from all causes [SMR=0.80, 90% confidence interval (CI): 0.78, 0.82], all cancers (SMR=0.85, 90% CI: 0.81, 0.89), and lung cancer (SMR=0.88, 90% CI: 0.82, 0.95) than expected based upon US mortality rates. The SMR for cancer of the pleura was 4.25 (90% CI: 1.99, 7.97) for men. The SMR for leukemia was greater than unity for monthly-paid (SMR=1.33, 90% CI: 0.88, 1.93) and hourly-paid (SMR=1.36, 90% CI: 1.02, 1.78) men. Female workers had fewer deaths from all causes (SMR=0.75, 90% CI: 0.69, 0.82) than expected, but more deaths than expected from cancer of the kidney (SMR=2.58, 90% CI: 1.21, 4.84) and skin (SMR=3.90, 90% CI: 2.11, 6.61). CONCLUSIONS: While the observed numbers of deaths in most categories of cause of death were less than expected, there are greater than expected numbers of deaths due to cancer of the pleura and leukemia, particularly among hourly-paid male workers. It is plausible that occupational hazards, including asbestos and ionizing radiation, contribute to these excesses. PMID- 17918228 TI - Absence of OLIG2 mutations in patients presenting with a severe Pelizaeus Merzbacher-like leukodystrophy associated with motor neuron dysfunction. PMID- 17918230 TI - Cross-shift peak expiratory flow changes are unassociated with respirable coal dust exposure among South African coal miners. AB - BACKGROUND: The objectives of this study were to determine whether cross-shift changes in peak expiratory flow rate (PEFR) were related to respirable dust exposure in South African coalminers. METHODS: Fifty workers were randomly selected from a cohort of 684 miners from 3 bituminous coalmines in Mpumalanga, South Africa. Peak expiratory efforts were measured prior to the commencement of the shift, and at the end of the shift on at least two occasions separated by at least 2 weeks, with full shift personal dust sampling being conducted on each occasion for each participant. Interviews were conducted, work histories were obtained and cumulative exposure estimates were constructed. Regression models examined the associations of cross-shift changes in PEFR with current and cumulative exposure, controlling for shift, smoking and past history of tuberculosis. RESULTS: There were marginal differences in cross-shift PEFR (ranging from 0.1 to 2 L/min). Linear regression analyses showed no association between cross-shift change in PEFR and current or cumulative exposure. The specific shift worked by participants in the study showed no effect. CONCLUSIONS: Our study showed no association between current respirable dust exposure and cross-shift changes in PEFR. There was a non-significant protective effect of cumulative dust exposure on the outcome, suggesting the presence of a "healthy worker survivor effect" in this data. PMID- 17918231 TI - On the job illness and injury resulting in lost work time among a national cohort of emergency medical services professionals. AB - BACKGROUND: The objective of this study was to estimate the prevalence and incidence of job-related illness or injury resulting in lost work time among a national cohort of Emergency Medical Services (EMS) professionals. Also, it was hypothesized that individual and work life characteristics were associated with the occurrence of illnesses or injury. METHODS: Data for this analysis were obtained from the Longitudinal Emergency Medical Technician Attributes and Demographics Study (LEADS), a prospective study of EMS professionals. The outcome variable of interest was self-reported absence from their EMS job due to an EMS work related illness or injury. The prevalence and incidence of injury with lost work time was estimated using cross-sectional and follow-up data. Multivariable logistic regression analyses were performed to determine if individual and work life characteristics were associated with occupational injury. RESULTS: The prevalence of job-related illness or injury with time away from work was estimated at 9.4%, while the 1-year incidence was estimated at 8.1 per 100 EMS providers. The results from the logistic regression model fit to follow-up data indicate that increasing call volume (OR=3.12 for very high vs. moderate, 95% CI 1.40-6.97), an urban work environment (OR=2.79, 95% CI 1.65-4.72) and a history of back problems (OR=1.72, 95% CI 1.06-2.78) were associated with reporting job related illness or injury. CONCLUSIONS: Results from this analysis provide estimates of the prevalence and incidence of on the job illness and injury resulting in lost work time among a national cohort of EMS professionals. PMID- 17918233 TI - Genetic association of neurotrophic tyrosine kinase receptor type 2 (NTRK2) With Alzheimer's disease. AB - Brain-derived neurotrophic factor (BDNF)/tyrosine receptor kinase (TRK) signaling pathway activates a wide range of downstream intracellular cascades, regulating neuronal development and plasticity, long-term potentiation, and apoptosis. The NTRK family encodes the receptors TRKA, TRKB, and TRKC, to which the neurotrophins, nerve growth factor (NGF), BDNF and neurotrophin-3 (NT-3) bind, respectively, with high affinity. Signaling through these receptors appears to be compromised in Alzheimer's disease (AD). This study is the most comprehensive investigation of genetic variants of NTRK2, and the first to show significant association between NTRK2 with AD. Fourteen single nucleotide polymorphisms (SNPs), located in 8 of 18 linkage disequilibrium (LD) blocks, were genotyped in 203 families with at least two AD affected siblings with mean age of onset (MAO) of 70.9 +/- 7.4 years and one unaffected sibling from the NIMH-ADGJ dataset. Family based association testing found no single SNP association, however, significant associations were found for two and three locus haplotypes (P = 0.012, P = 0.009, respectively) containing SNPs rsl624327, rsl443445, and rs378645. These SNPs are located in areas of the gene containing sequences that could be involved in alternative splicing and/or regulation of NTRK2. Our results suggest that NTRK2 may be a genetic susceptibility gene contributing to AD pathology. PMID- 17918232 TI - Alpha-synuclein promoter haplotypes and dementia in Parkinson's disease. AB - Dementia is a common complication of Parkinson's disease (PD). It correlates significantly with the presence of cortical, limbic or nigral Lewy bodies, mainly constituted of alpha-synuclein. Mutations of the alpha-synuclein gene (SNCA) have been linked to rare familial forms of PD, while association studies on the promoter polymorphisms have given conflicting results in sporadic patients. We have performed a case control study to investigate whether genetic variability in the promoter of the alpha-synuclein gene could predispose to dementia in PD. A total of 114 demented patients and 114 non-demented patients with sporadic PD were included in the study. Six polymorphic loci (including the Rep1 microsatellite) in the promoter of the SNCA gene were examined. Each marker, taken individually, did not show association to dementia and no significant differences were observed in the inferred haplotype frequencies of demented and non-demented patients. Our data suggest the lack of involvement of the SNCA promoter in the pathogenesis of dementia in PD. Further studies in other populations are needed to confirm these results. PMID- 17918234 TI - The monoamine oxidase B gene exhibits significant association to ADHD. AB - Attention deficit hyperactivity disorder (ADHD) is a common neuropsychiatric condition with strong genetic basis. Recent work in China indicated that ADHD may be linked to Xp1-2 in the Han Chinese population. The gene encoding monoamine oxidase B (MAOB), the main enzyme degrading dopamine in the human brain, is located in this region. The current study sequenced the exons and the 5' and 3' flanking regions of the MAOB gene and found four common variants including 2276C>T and 2327C>T in exon 15, rs1799836 in intron 13 and rs1040399 in 3'-UTR. We assessed the association of these variants with ADHD in 548 trios collected from 468 males and 80 females probands. TDT analysis showed that alleles of each polymorphism were preferentially transmitted to probands (rs1799836, P = 3.28E 15; rs1040399, P = 1.87E-6; 2276T>C or 2327T>C, P = 2.20E-6) and haplotype-based TDT analyses also found distorted transmission. In conclusion, this study provides the strongest evidence for the involvement of MAOB gene in the etiology of ADHD to date, at least in Han Chinese population. PMID- 17918235 TI - Experience-dependent plasticity in the mushroom bodies of the solitary bee Osmia lignaria (Megachilidae). AB - All members of the solitary bee species Osmia lignaria (the orchard bee) forage upon emergence from their natal nest cell. Conversely, in the honey bee, days-to weeks of socially regulated behavioral development precede the onset of foraging. The social honey bee's behavioral transition to foraging is accompanied by neuroanatomical changes in the mushroom bodies, a region of the insect brain implicated in learning. If these changes were general adaptations to foraging, they should also occur in the solitary orchard bee. Using unbiased stereological methods, we estimated the volume of the major compartments of the mushroom bodies, the neuropil and Kenyon cell body region, in adult orchard bees. We compared the mushroom bodies of recently emerged bees with mature bees that had extensive foraging experience. To separate effects of general maturation from field foraging, some orchard bees were confined to a cage indoors. The mushroom body neuropil of experienced field foragers was significantly greater than that of both recently emerged and mature caged orchard bees, suggesting that, like the honey bee, this increase is driven by outdoor foraging experience. Unlike the honey bee, where increases in the ratio of neuropil to Kenyon cell region occur in the worker after emerging from the hive cell, the orchard bee emerged from the natal nest cell with a ratio that did not change with maturation and was comparable to honey-bee foragers. These results suggest that a common developmental endpoint may be reached via different development paths in social and solitary species of foraging bees. PMID- 17918236 TI - Association study of the brain-derived neurotropic factor (BDNF) gene in attention deficit hyperactivity disorder. AB - Attention deficit hyperactivity disorder (ADHD) is a prevalent neurodevelopmental childhood psychiatric disorder. Brain-derived neurotropic factor (BDNF) has been suggested to play a role in the pathogenesis of ADHD and two family-based association studies demonstrated an association of BDNF polymorphisms with ADHD. The aim of the current study was to investigate the BDNF gene for association with ADHD in a large sample of families from Toronto. The transmission of three polymorphisms of the BDNF gene (rs6265, rs11030104, and rs2049046) was examined in 266 nuclear families ascertained through a proband with ADHD (315 affected children) using the transmission/disequilibrium test (TDT). In addition, we conducted quantitative analysis to assess the relationship between these marker alleles and the symptom dimensions of ADHD (inattention and hyperactivity/impulsivity) and cognitive measures of working memory. None of the individual marker alleles showed significant evidence of association with ADHD, dimensional symptom scores, or working memory ability in our sample of ADHD families. There was no significant evidence for biased transmission of individual haplotypes with frequency >10% (global chi2 for these three haplotypes: chi2 = 6.349, df = 3, P = 0.096). One uncommon haplotype (A-G-G; frequency 2.2%) showed a significant association with ADHD in the categorical (chi2 = 5.293, df = 1, P = 0.021) and quantitative analyses (parents' rated inattention: Z = -2.504, P = 0.012; and hyperactivity/impulsivity: Z = -2.651, P = 0.008). These results should be interpreted cautiously, however, because of the low haplotype frequency. In light of the evidence for an involvement of BDNF in ADHD, further analysis of the BDNF gene in ADHD is warranted. PMID- 17918237 TI - 14-3-3epsilon Expression is induced during the critical period of thermal control establishment. AB - Signal-transduction mechanisms leading to neuronal plasticity involve a series of phosphorylation steps, which lead to transcription and de-novo protein synthesis. However, it is not clear how the specificity of the signal transduction achieved. Here, we evaluated the role of 14-3-3epsilon, a chaperone of phosphorylation and cellular-localization determination, in thermal control establishment, which represents hypothalamic plasticity, in chicks. As with other sensory mechanisms, there is a critical period in the development of temperature control. Neuroanatomically, body temperature is balanced by the preoptic anterior hypothalamus (PO/AH) and controlled by thermosensitive neurons. Hot or cold exposure during the critical period of temperature control development causes a plastic change in the ratio between hot- and cold-sensitive cells and can modulate temperature tolerance throughout life. It has been found that both mRNA and protein of isoform 14-3-3epsilon, but not other 14-3-3 isoforms, are induced during hot and cold conditioning in the PO/AH, with a peak at 24 h of conditioning. To determine whether 14-3-3epsilon has a general role in neuronal plasticity, we checked its expression after passive avoidance learning in the relevant neuroanatomical areas, i.e. the mesopallium intermediomediale and the striatum mediale. It was established that in these areas its expression was not altered after learning. The present study suggests that the correlative induction of both 14-3-3epsilon mRNA and protein many hours after learning is involved in plasticity in the hypothalamus but not in passive avoidance-related plasticity, which might indicate that specificity in the signal-transduction mechanism is at least partially mediated by 14-3-3. PMID- 17918238 TI - The zebrafish ennui behavioral mutation disrupts acetylcholine receptor localization and motor axon stability. AB - The zebrafish ennui mutation was identified from a mutagenesis screen for defects in early behavior. Homozygous ennui embryos swam more slowly than wild-type siblings but normal swimming recovered during larval stages and homozygous mutants survived until adulthood. Electrophysiological recordings from motoneurons and muscles suggested that the motor output of the CNS following mechanosensory stimulation was normal in ennui, but the synaptic currents at the neuromuscular junction were significantly reduced. Analysis of acetylcholine receptors (AChRs) in ennui muscles showed a marked reduction in the size of synaptic clusters and their aberrant localization at the myotome segment borders of fast twitch muscle. Prepatterned, nerve-independent AChR clusters appeared normal in mutant embryos and dispersed upon outgrowth of motor axons onto the muscles. Genetic mosaic analysis showed that ennui is required cell autonomously in muscle fibers for normal synaptic localization of AChRs. Furthermore, exogenous agrin failed to induce AChR aggregation, suggesting that ennui is crucial for agrin function. Finally, motor axons branched more extensively in ennui fast twitch muscles especially in the region of the myotome borders. These results suggest that ennui is important for nerve-dependent AChR clustering and the stability of axon growth. PMID- 17918239 TI - The functional polymorphism of the hemoglobin-binding protein haptoglobin influences susceptibility to idiopathic Parkinson's disease. AB - Oxidative stress and iron have been widely implicated in the etiology of Parkinson's disease (PD). Hemoglobin is the richest source of iron in the body. The human Haptoglobin (Hp) protein is a plasma alpha-2 glycoprotein that removes free Hemoglobin from the circulation and tissues and is important in protection from oxidative stress, in immune system regulation, and angiogenesis. A common genetic polymorphism of Hp exists in the population, where the Hp 1-1, Hp 2-1, and Hp 2-2 forms exhibit profound functional differences. In this study, the Hp genotype corresponding to phenotypes Hp 1-1, 2-1 and 2-2 was determined in 312 idiopathic PD patients and 420 normal control subjects. A significant increase in the number of subjects carrying the Hp 2-1 genotype was present among PD patients. The distribution of Hp genotypes among PD patients (16.0% Hp 1-1, 56.4% Hp 2-1, 27.6% Hp 2-2) was significantly different from the distribution in controls (15.2% Hp 1-1, 48.1% Hp 2-1, 36.7% Hp 2-2) (chi(2) = 6.99, P = 0.030). The odds ratios for PD risk for Hp 2-1 and Hp 1-1 versus Hp 2-2 genotype were 1.51 (1.07-2.12) and 1.36 (0.86-2.15), respectively. Overall, the association of Hp-1 allele with PD resulted stronger among subjects who were never-smokers as compared to ever-smokers. Also, among ever-smokers, Hp genotypes were significantly associated with PD only among women, but not men, indicating the presence of a gene x gender x smoking interaction. To our knowledge, this is the first study that investigates the association of Hp genotypes with the risk of PD. PMID- 17918240 TI - Distribution of neurotrophin-3 during the ontogeny and regeneration of the lizard (Gallotia galloti) visual system. AB - We have previously described the spontaneous regeneration of retinal ganglion cell axons after optic nerve (ON) transection in the adult Gallotia galloti. As neurotrophin-3 (NT-3) is involved in neuronal differentiation, survival and synaptic plasticity, we performed a comparative immunohistochemical study of NT-3 during the ontogeny and regeneration (after 0.5, 1, 3, 6, 9, and 12 months postlesion) of the lizard visual system to reveal its distribution and changes during these events. For characterization of NT-3(+) cells, we performed double labelings using the neuronal markers HuC-D, Pax6 and parvalbumin (Parv), the microglial marker tomato lectin or Lycopersicon esculentum agglutinin (LEA), and the astroglial markers vimentin (Vim) and glial fibrillary acidic protein (GFAP). Subpopulations of retinal and tectal neurons were NT-3(+) from early embryonic stages to adulthood. Nerve fibers within the retinal nerve fiber layer, both plexiform layers and the retinorecipient layers in the optic tectum (OT) were also stained. In addition, NT-3(+)/GFAP(+) and NT-3(+)/Vim(+) astrocytes were detected in the ON, chiasm and optic tract in postnatal and adult lizards. At 1 month postlesion, abundant NT-3(+)/GFAP(+) astrocytes and NT-3(-)/LEA(+) microglia/macrophages were stained in the lesioned ON, whereas NT-3 became downregulated in the experimental retina and OT. Interestingly, at 9 and 12 months postlesion, the staining in the experimental retina resembled that in control animals, whereas bundles of putative regrown fibers showed a disorganized staining pattern in the OT. Altogether, we demonstrate that NT-3 is widely distributed in the lizard visual system and its changes after ON transection might be permissive for the successful axonal regrowth. PMID- 17918241 TI - Arachidonic acid as a retrograde signal controlling growth and dynamics of retinotectal arbors. AB - In the developing visual system, correlated presynaptic activity between neighboring retinal ganglion cells (RGC) stabilizes retinotopic synapses via a postsynaptic NMDAR (N-methyl-D-aspartate receptor)-dependent mechanism. Blocking NMDARs makes individual axonal arbors larger, which underlies an unsharpened map, and also increases branch turnover, as if a stabilizing factor from the postsynaptic partner is no longer released. Arachidonic acid (AA), a candidate retrograde stabilizing factor, is released by cytoplasmic phospholipase A2 (cPLA2) after Ca(2+) entry through activated NMDARs, and can activate presynaptic protein kinase C to phosphorylate various substrates such as GAP43 to regulate cytoskeletal dynamics. To test the role of cPLA2 in the retinotectal system of developing zebrafish, we first used PED6, a fluorescent reporter of cPLA2 activity, to show that 1-3 min of strobe flashes activated tectal cPLA2 by an NMDAR-dependent mechanism. Second, we imaged the dynamic growth of retinal arbors during both local inhibition of tectal cPLA2 by a pharmacological inhibitor, arachidonic tri-fluoromethylketone, and its suppression by antisense oligonucleotides (both injected intraventricularly). Both methods produced larger arbors and faster branch dynamics as occurs with blocking NMDARs. In contrast, intraocular suppression of retinal cPLA2 with large doses of antisense oligos produced none of the effects of tectal cPLA2 inhibition. Finally, if AA is the retrograde messenger, the application of exogenous AA to the tectum should reverse the increased branch turnover caused by blocking either NMDARs or cPLA2. In both cases, intraventricular injection of AA stabilized the overall branch dynamics, bringing rates down below the normal values. The results suggest that AA generated postsynaptically by cPLA2 downstream of Ca(2+) entry through NMDARs acts as a retrograde signal to regulate the dynamic growth of retinal arbors. PMID- 17918242 TI - ADH7 variation modulates extraversion and conscientiousness in substance dependent subjects. AB - Human personality traits have been closely linked to substance dependence (SD), and are partially genetically determined. Recently, associations between alcohol dehydrogenase 7 (ADH7) and SD have been reported, which led us to investigate the relationship between ADH7 variation and personality traits. We assessed dimensions of the five-factor model of personality and genotyped 4 ADH7 markers and 38 unlinked ancestry-informative markers in 244 subjects with SD [178 European-Americans (EAs) and 66 African-Americans (AAs)] and 293 healthy subjects (253 EAs and 40 AAs). The relationships between ADH7 markers and personality traits were comprehensively examined using multivariate analysis of covariance (MANCOVA), and then decomposed by Roy Bargmann Stepdown analysis of covariance (ANCOVA). Generally, older individuals, AAs, and males had significantly lower personality scores (4.7 x 10(-5) < or = P < or = 0.032), as reported previously. In SD subjects, Extraversion was most significantly associated with ADH7 haplotypes (3.7 x 10(-4) < or = P < or = 0.001), diplotypes (0.007 < or = P < or = 0.012), and genotypes (P = 0.001), followed by Conscientiousness (0.005 < or = P < or = 0.033). The contributory haplotype and diplotypes contained the alleles and genotypes of rs284786 (SNP1) and rs1154470 (SNP4). In healthy subjects, other personality factors (except Extraversion) were associated with ADH7 diplotypes (0.005 < or = P < or = 0.016) and genotypes (0.002 < or = P < or = 0.052). Some of the gene effects on personality factors were modified by sex. The present study demonstrated that the ADH7 variation may contribute to the genetic component of variation in personality traits, with the risk for SD and personality traits being partially shared. PMID- 17918243 TI - Exogenous Hsc70, but not thermal preconditioning, confers protection to motoneurons subjected to oxidative stress. AB - Proper sensing of stress and the initiation of the stress response are critical to maintaining cell viability in response to noxious stimuli. Induction of the stress response prior to the exposure of a lethal stress (preconditioning) can be protective. Heat shock proteins (Hsps), the main products of the stress response, are considered to be responsible for this protective effect. Most cells readily initiate a stress response, but some neuronal phenotypes, including motoneurons (MNs), have a diminished capacity to do so. We have found that, given a proper stimulus, MNs can execute a heat stress response; but, it does not protect them from death caused by hydrogen peroxide (H(2)O(2)) induced oxidative stress, despite inhibiting H(2)O(2)-induced caspase activation. Conversely, we demonstrate that incubation with the heat shock cognate 70 (Hsc70) protein prior to oxidative insult can protect MNs from oxidative stress. This survival promoting effect may be mediated through the substrate binding domain (SBD) of Hsc70. Our data suggest that stress preconditioning may not be beneficial to MNs, but that pharmacological interventions and alternative means of acquiring components of the stress response are an effective means of ameliorating lethal stress in MNs and may be potentially useful therapeutically in preventing pathological MN loss. PMID- 17918245 TI - Role for para sodium channel gene 3' UTR in the modification of Drosophila seizure susceptibility. AB - Voltage-gated sodium channel genes are highly regulated at the level of transcription or translation. In this study, we have utilized the combination of genetic, electrophysiological, and molecular analyses to identify a 7-kb 3' untranslated region (UTR) of the Drosophila para sodium channel gene. Disruption of this segment by P-element insertion causes reduction of para primary transcript, but not Rbp2 transcripts. The identification of this novel 3'-UTR is based on a P-insertion mutation called para(JS1), which was isolated from a P element mutagenesis screen for seizure suppressors in a Drosophila model of epilepsy. The para(JS1) mutation was identified 6845 bp downstream of the para gene, which resides in an intergenic region that lies between para and Rbp2 (RNA binding protein 2) genes. Interestingly, reverse-transcription PCR showed that the region containing para(JS1) is substantially transcribed and this transcribed region is associated with para coding region. We discussed possible mechanisms of how reduced transcription of the para gene or alterations in sodium channel subunit composition might be indicated by the para(JS1) mutation and implications for para 3' UTR function. PMID- 17918244 TI - Functional delay of myelination of auditory delay lines in the nucleus laminaris of the barn owl. AB - In the barn owl, maps of interaural time difference (ITD) are created in the nucleus laminaris (NL) by interdigitating axons that act as delay lines. Adult delay line axons are myelinated, and this myelination is timely, coinciding with the attainment of adult head size, and stable ITD cues. The proximal portions of the axons become myelinated in late embryonic life, but the delay line portions of the axon in NL remain unmyelinated until the first postnatal week. Myelination of the delay lines peaks at the third week posthatch, and myelinating oligodendrocyte density approaches adult levels by one month, when the head reaches its adult width. Migration of oligodendrocyte progenitors into NL and the subsequent onset of myelination may be restricted by a glial barrier in late embryonic stages and the first posthatch week, since the loss of tenascin-C immunoreactivity in NL is correlated with oligodendrocyte progenitor migration into NL. PMID- 17918246 TI - Effect of oleic and linoleic acids on the inflammatory phase of wound healing in rats. AB - Inflammation is a crucial step for the wound healing process. The effect of linoleic and oleic acids on the inflammatory response of the skin during the healing process and on the release of pro-inflammatory cytokines by rat neutrophils in vitro was investigated. A wound in the dorsal surface of adult rats was performed and fatty acids were then topically administered. Both oleic and linoleic acids increased the wound healing tissue mass. The total protein and DNA contents of the wounds were increased by the treatment with linoleic acid. The treatments with oleic and linoleic acids did not affect vascular permeability. However, the number of neutrophils in the wounded area and air pouches was increased and the thickness of the necrotic cell layer edge around the wound was decreased. A dose-dependent increase in vascular endothelial growth factor-alpha (VEGF-alpha) and interleukin-1beta (IL-1beta) by neutrophils incubated in the presence of oleic and linoleic acid was observed. Oleic acid was able to stimulate also the production of cytokine-induced neutrophil chemoattractant in inflammation 2 alpha/beta (CINC-2alpha/beta). This pro inflammatory effect of oleic and linoleic acids may speed up the wound healing process. PMID- 17918247 TI - Minimizing risk associated with elderly liver donors by matching to preferred recipients. AB - Elderly liver donors (ELDs) represent a possible expansion of the donor pool, although there is great reluctance to use ELDs because of reports that increasing donor age predicts graft loss and patient death. The goal of this study was to identify a subgroup of recipients who would be least affected by increased donor age and thus best suited to receive grafts from ELDs. A national registry of deceased donor liver transplants from 2002-2005 was analyzed. ELDs aged 70-92 (n = 1043) were compared with average liver donors (ALDs) aged 18-69 (n = 15,878) and ideal liver donors (ILDs) aged 18-39 (n = 6842). Recipient factors that modified the effect of donor age on outcomes were identified via interaction term analysis. Outcomes in recipient subgroups were compared using Kaplan-Meier survival analysis. Recipients preferred for ELD transplants were determined to be first-time recipients over the age of 45 with body mass index <35, non-status 1 registration, cold ischemic time <8 hours, and either hepatocellular carcinoma or an indication for transplantation other than hepatitis C. In preferred recipients, there were no differences in outcomes when ELD livers were used (3 year graft survival: ELD 75%, ALD 75%, ILD 77%, P > 0.1; 3-year patient survival: ELD 81%, ALD 80%, ILD 81%, P > 0.1). In contrast, there were significantly worse outcomes when ELD livers were used in nonpreferred recipients (3-year graft survival: ELD 50%, ALD 71%, ILD 75%, P < 0.001; 3-year patient survival: ELD 64%, ALD 77%, ILD 80%, P < 0.001). CONCLUSION: The risks of ELDs can be substantially minimized by appropriate recipient selection. PMID- 17918248 TI - Arterial compliance and retinal vascular caliber in cerebrovascular disease. AB - OBJECTIVE: Arterial stiffness is a newly recognized risk factor for stroke. Whether this is mediated by small- or large-artery disease is unknown. In this study, we examined the relationship between arterial stiffness and retinal vascular caliber. METHODS: A cross-sectional, population-based study of 5,731 participants (aged 45-84 years) who were free of clinical cardiovascular disease (the Multi-Ethnic Study of Atherosclerosis). Retinal arteriolar and venular calibers were measured from retinal photographs according to standardized protocols. Small- and large-artery compliance was determined from pulse contour analysis. RESULTS: After adjusting for age, sex, blood pressure, diabetes, smoking, lipid profile, and other risk factors, reduced large-artery compliance was associated with smaller retinal arteriolar caliber; each standard deviation decrease in large-artery compliance was associated with a 0.70 microm (p = 0.002) decrease in retinal arteriolar caliber. After adjusting for the same set of risk factors, reduced small-artery compliance was associated with wider retinal venular caliber; each standard deviation decrease in small artery compliance was associated with a 1.43 microm (p = 0.001) increase in retinal venular caliber. These associations remained significant after further adjustments for large vessel atherosclerosis (carotid intima-media thickness, coronary artery calcium, and ankle-arm index). INTERPRETATION: Reduced arterial wall compliance in large arterial beds is associated with retinal arteriolar narrowing, whereas reduced arterial wall compliance in small arterial beds is associated with retinal venular widening. These data may provide further insights into the effects of altered arterial stiffness on the cerebral microcirculation. PMID- 17918249 TI - Fetal hemoglobin in sickle cell anemia: Bayesian modeling of genetic associations. AB - We genotyped single nucleotide polymorphisms (SNPs) in: (1) the beta-globin gene like cluster, (2) quantitative trait loci (QTL) previously associated with fetal hemoglobin (HbF) concentration on chromosomes 6q, 8q, and Xp, and (3) candidate genes that could effect HbF levels, in sickle cell anemia subjects. HbF concentration was modeled as a continuous variable with values in a finite interval using a novel Bayesian approach. We first tested the associations of SNPs with HbF in a group of 1,518 adults and children (CSSCD study), and validated the results in a second independent group of 211 adults (MSH study). In subjects aged >or=24 years, 5 SNPs in TOX (8q12.1), 2 SNPs in the beta-globin gene-like cluster, 2 SNPs in the Xp QTL, and 1 SNP in chromosome 15q22 were associated with HbF in the CSSCD and also validated in the MSH. Four other SNPs in 15q22 were associated with HbF only in the larger CSSCD data. When patients aged <24 years in the CSSCD were examined, additional genes, including 4 with roles in nitric oxide metabolism, were associated with HbF level. These studies confirm prior analyses using traditional analytical approaches showing associations of SNPs in TOX, GPM6B, and the beta-globin gene-like cluster with HbF levels. We also identified an additional candidate regulatory region in chromosome 15q22 that is associated with HbF level. By stratifying patients by age, our results also suggest that different genes might modulate the rate of decline of HbF and the final level of HbF levels in sickle cell anemia. PMID- 17918250 TI - Superficial venous thrombosis associated with congenital absence of the inferior vena cava and previous episode of deep venous thrombosis. AB - Congenital malformations of the inferior vena cava (IVC) are uncommon and may be associated with an increased risk of venous thrombosis. We report the case of a man with congenital absence of the IVC and remote history of deep venous thrombosis who now presents with severe abdominal wall superficial thrombophlebitis. To our knowledge, this is the first report of a patient with IVC absence who has developed both deep and superficial venous thromboses. PMID- 17918251 TI - Late onset of autoimmune hemolytic anemia and pure red cell aplasia after allogeneic hematopoietic stem cell transplantation using in vivo alemtuzumab. AB - Hemolytic anemia and pure red cell aplasia (PRCA) after allogeneic hematopoietic stem cell transplantation (HSCT) have been reported to be mainly related to ABO incompatibility between donor and recipient. Autoimmune hemolytic anemia (AIHA) without ABO-incompatibility has been also reported after allogeneic HSCT, especially with T-cell depletion. However, optimal management of AIHA or PRCA remains unclear. A 54-year-old male with myelodysplastic syndrome (MDS) underwent haploidentical human leukocyte antigen-mismatched HSCT using in vivo alemtuzumab and developed AIHA and PRCA simultaneously 15 months after transplantation, following the administration of cidofovir and probenecid for persistent cytomegalovirus (CMV) antigenemia and retinitis. AIHA was successfully treated with rituximab, and subsequently PRCA with cyclosporine without relapse of MDS or recurrence of CMV infection. The clinical course suggested that AIHA was mainly caused by humoral immune response, while PRCA was mainly caused by cell-mediated immune response in this patient, although these immune responses might be related to each other. PMID- 17918252 TI - Soluble TWEAK is markedly elevated in hemophagocytic lymphohistiocytosis. AB - Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is a newly identified monocyte derived cytokine, which has weak apoptosis inducing function against sensitive tumor cell lines in vitro. Also, TWEAK has been reported to have proangiogenic and proinflammatory activities in vivo. However, its functions in pathological situation remain to be elucidated. Here, we analyzed soluble TWEAK in serum of 24 patients with hemophagocytic lymphohistiocytosis (HLH) in combination with interferon-gamma (IFN-gamma) and killer-specific secretory protein of 37 kDa (Ksp37). Soluble TWEAK was not detected in serum of healthy individuals. Soluble TWEAK was markedly elevated in all six primary HLH patients and 12 of 18 secondary HLH patients. Serum IFN-gamma, which is an only known mediator to stimulate TWEAK production in monocyte in vitro, was not elevated despite elevated serum TWEAK in three of six primary HLH patients, although IFN gamma was markedly elevated in other cases. Ksp37 was only slightly increased in HLH patients. These results indicate that TWEAK may be involved in pathogenesis of HLH and is useful as a clinical marker. PMID- 17918253 TI - Decreased incidence of acute graft-versus-host disease by continuous infusion of cyclosporine with a higher target blood level. AB - Cyclosporine A (CsA) is the mainstay of pharmacologic prevention of acute graft versus-host disease (GVHD). We previously reported that continuous infusion of CsA with a target blood level between 250 and 400 ng/ml significantly increased the incidence of acute GVHD compared to twice-daily infusion with a target trough level between 150 and 300 ng/ml. Thus, we raised the target level of CsA continuous infusion to 450-550 ng/ml. We treated 33 patients with the higher target level (CsA500) and compared the efficacy and toxicity with those in the 33 historical control patients (CsA300 group). Other transplantation procedures were not changed. The patients' characteristics were equivalent. The average CsA concentration was adjusted around 500 ng/ml and the actual daily dose was maintained at the initial dose (CsA 3mg/kg/day). Toxicities were equivalently observed among the two groups. The incidence of grades II-IV acute GVHD was significantly lower in the CsA500 group (27 vs. 52%, P = 0.033). The target level of CsA was identified as an independent significant risk factor for grades II-IV acute GVHD (P = 0.039), adjusted for the presence of HLA mismatch. The incidence of chronic GVHD was also decreased in the CsA500 group (47 vs. 73%, P = 0.016). We conclude that the toxicity of the continuous CsA infusion with a target level of 450-550 ng/ml is acceptable and the efficacy to prevent acute GVHD is significant. A larger comparative study is warranted to confirm these findings. PMID- 17918255 TI - Testing for recombinant erythropoietin. AB - Erythropoietin (Epo) is a glycoprotein hormone that promotes the production of red blood cells. Recombinant human Epo (rhEpo) is illicitly used to improve performance in endurance sports. Doping in sports is discouraged by the screening of athletes for rhEpo. Both direct tests (indicating the presence of exogeneous Epo isoforms) and indirect tests (indicating hematological changes induced by exogenous Epo administration) can be used for Epo detection. At present, the test adopted by the World Anti Doping Agency is based on a combination of isoelectric focusing and double immunoblotting, and distinguishes between endogenous and rhEpo. However, the adopted monoclonal anti-Epo antibodies are not monospecific. Therefore, the test can occasionally lead to the false-positive detection of rhEpo (epoetin-beta) in post-exercise, protein-rich urine, or in case of contamination of the sample with microorganisms. An improved preanalytical care may counteract a lot of these problems. Adaptation of the criteria may be helpful to further refine direct Epo testing. Indirect tests have the disadvantage that they require blood instead of urine samples, but they can be applied to detect a broader range of performance improving techniques which are illicitly used in sports. PMID- 17918256 TI - Nodular lymphocyte predominant Hodgkin lymphoma at atypical locations may be associated with increased numbers of large cells and a diffuse histologic component. AB - Nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) typically affects predictable lymph node groups with excellent treatment outcomes, but cases with a diffuse histologic pattern are associated with recurrence and rarely, cases will transform to diffuse large B-cell lymphoma. Although increased numbers of large cells has not been associated with poor prognosis, transformation is thought to histologically progress through a stage distinguished by increasing numbers of large atypical B-cells. From 55 cases of NLPHL, we describe a possible subset of NLPHL occurring in older individuals at atypical sites, associated with increased numbers of large cells, a diffuse histologic component, and expression of Bcl-2. PMID- 17918257 TI - Molecular identification of COL6A3-CSF1 fusion transcripts in tenosynovial giant cell tumors. AB - Tenosynovial giant cell tumors (TGCTs) are benign lesions of the tendon sheaths that primarily affect the fingers, ankles, or feet. Cytogenetic data have shown that 1p13 is most frequently involved in structural aberrations and that 2q37 is its most common translocation partner. The genes involved in the translocation t(1;2)(p13;q37) were recently identified: the colony-stimulating factor-1 (CSF1 or M-CSF1) at 1p13 and the collagen type VI alpha-3 (COL6A3) at 2q37. Based upon the suggestion that a fusion of these genes through the translocation would result in overexpression of CSF1 due to a strong COL6A3 promoter, we performed RT PCR on six TGCT cases with t(1;2) to search for a putative COL6A3-CSF1 fusion gene. Such fusion transcripts were detected in three cases of which one was an in frame fusion. In all cases, however, the breakpoints in CSF1 appeared downstream of exon 5, indicating that the amino-terminal part of CSF1, which interacts with its receptor CSF1R, is not encoded by the the chimeric transcripts we identified. The pathogenetic mechanism of these chimeric transcripts is therefore unclear. PMID- 17918258 TI - Primary salivary gland-type lung cancer: spectrum of clinical presentation, histopathologic and prognostic factors. AB - BACKGROUND: Primary salivary-type lung cancers are rare tumors that include adenoid cystic carcinoma (ACC) and mucoepidermoid carcinoma (MEC). The clinicopathologic profiles, symptoms on presentation, and long-term outcomes of patients with ACC and MEC as an overall group have not been defined recently. METHODS: In this study, the authors analyzed clinical outcome data from 62 patients who presented with a diagnosis of primary salivary-type lung cancer at the Mayo Clinic (Rochester, Minn) from 1972 to 2002. RESULTS: The median age at diagnosis for patients with MEC was 40 years (range, 6-78 years); and, for patients ACC, the median age at diagnosis was 54 years (range, 21-76 years). ACC was observed more frequently among women and girls. The main presenting symptom for both tumors was cough (70%), followed by dyspnea (51.7%), wheezing (38.3%), obstructive pneumonia (30%), hemoptysis (28.3%), and fever (16.7%). Tissues were available for review from all patients. Among the ACC tumors, 29 (74.4%) were cribriform, 7 (17.9%) were tubular, and 3 (7.7%) were the solid type. Most MEC tumors (65%) were intermediate grade (grade 2), and 30% were low grade (grade 1). Most salivary-type lung cancers presented in the trachea, in the carina, or in a main stem bronchus (70.7%). This location was observed more often (82.5%) for ACC tumors compared with MEC tumors (44.4%). Involvement of the lymph nodes was observed in 20% of patients and was more common among the patients with ACC (30.8%). Distant metastases were observed in 30.4% of the patients (15 patients in the ACC group [40.5%] compared with only 2 patients in the MEC group [10.5%]; P = .03). For patients who underwent complete surgical resection, the 3-, 5-, and 10-year survival rates were 82%, 70%, and 63%, respectively. The survival rates for surgical MEC patients were 94% at 3 years and 87% at both 5 years and 10 years. For surgical ACC patients, the survival rates were 73%, 57%, and 45% at 3, 5, and 10 years, respectively. The survival rate for patients with ACC who did not undergo surgery was 74% at 3 years, 53% at 5 years, and 31% at 10 years. The difference in survival between surgical and nonsurgical patients was statistically significant (P < .01). CONCLUSIONS: Patients with MEC and ACC frequently have a good long-term prognosis but do not always have indolent diseases. Local recurrence is likely if complete surgical resection is not achieved. ACC has a higher likelihood than MEC to metastasize. Overall, patients with MEC survive better than patients with ACC. PMID- 17918259 TI - Radiation therapy is associated with improved survival in patients with pancreatic adenocarcinoma: results of a study from the Surveillance, Epidemiology, and End Results (SEER) registry data. AB - BACKGROUND: The role of adjuvant radiation therapy in pancreatic cancer is controversial. For the current study, the authors evaluated the effect of preoperative and postoperative radiation therapy on survival in patients with pancreatic adenocarcinoma. METHODS: The analysis included 3008 patients who were reported to the Surveillance, Epidemiology, and End Results registry of the National Cancer Institute from 1988 to 2002 who had adenocarcinoma of the pancreas and who underwent cancer-directed surgery. A retrospective analysis of overall survival and cancer-specific survival for these patients was performed using the Kaplan-Meier method. Comparative risks of mortality were evaluated by using multivariate-adjusted Cox regression models. RESULTS: Of 3008 patients, 1267 (42%) received radiation therapy. Overall survival improved significantly in patients who received radiation therapy, with a median survival of 17 months and a 5-year overall survival rate of 13% in patients who received radiation compared with 12 months and 9.7%, respectively, for patients who did not receive radiation therapy (P < .0001). On multivariate analysis, radiation therapy was associated with improvement in overall survival in patients who had direct extension beyond the pancreas and/or regional lymph node involvement (P < .01) but not in patients with T1-T2N0M0 disease (P > .05). Radiation therapy was associated with improvement in cause-specific survival in patients who had regional lymph node involvement (P < .02) but not in patients who had T1-2N0M0 disease or direct extension beyond the pancreas without lymph node involvement (P > .05). Differences in overall and cause-specific survival among patients who received preoperative versus postoperative radiation therapy did not reach statistical significance. CONCLUSIONS: Radiation therapy was associated with improved survival compared with cancer-directed surgery without radiation in patients with adenocarcinoma of the pancreas. PMID- 17918260 TI - Challenges in the recruitment of adolescents and young adults to cancer clinical trials. AB - The adolescent and young adult (AYA) oncology population has seen inferior progress in cancer survival compared with younger children and older adults over the past 25 years. Previously, AYAs had the best survival rates due to the prevalence of highly curable diseases including Hodgkin lymphoma and germ cell tumors, yet today AYAs have inferior survival rates to children and some adult cohorts. Survival rates are particularly poor for AYA-specific diseases such as sarcomas. Research involving children and adults diagnosed with common malignancies such as acute lymphoblastic leukemia has resulted in improved survival rates. However, AYAs have not directly benefited from such research due to low rates of access to and accrual on clinical trials. AYAs are less likely to have insurance or access to healthcare, are more likely to see providers who are not part of research institutions, and are less likely to be referred to or to join clinical trials, all of which may contribute to worse outcomes. Few clinical trials target AYA-specific diseases, leading to little information regarding how these diseases behave and what role the host plays. Tumor samples for this population are underrepresented in national tumor banks. Coupled with the need for more clinical trials that focus on AYA-specific cancers, better collaboration between adult and pediatric cooperative groups as well as increased education among community oncologists and primary care providers will be needed to enhance participation in clinical trials with the goal to increase survival and improve quality of that survival. PMID- 17918261 TI - Haptoglobin and posttranslational glycan-modified derivatives as serum biomarkers for the diagnosis of nonsmall cell lung cancer. AB - BACKGROUND: The purpose was to evaluate the clinical utility of serum haptoglobin (Hp) and posttranslational glycan modifications of Hp for the diagnosis of nonsmall cell lung cancer (NSCLC). METHODS: Serum proteins from patients with a new diagnosis of NSCLC and age- and sex-matched controls without cancer were compared using 2-dimensional difference gel electrophoresis (2D-DIGE). Four of the differentially expressed gel spots were identified as the beta chain of Hp. Immunoblots confirmed sialyl and fucosyl group posttranslational modifications (PTMs) of Hp. Serum enzyme-linked immunosorbent assays (ELISAs) for total Hp, sialylated Hp (SAHp), and fucosylated Hp (FHp) were designed, and levels of each were measured in an independent sample set of 74 patients. Receiver operating characteristic (ROC) analysis assessed the clinical diagnostic utility of each marker. RESULTS: Statistically significant differences between lung cancer patients and matched controls were found by ELISA for Hp (P < .002), SAHp (P < .001), and FHp (P < .04). ROC analysis determined an area under the curve (AUC) of 0.754 for Hp, 0.740 for SAHp, and 0.794 for FHp. In addition, serum concentrations correlated with stage; Hp (r = 0.388; P = .018), SAHp (r = 0.300; P = .072), and FHp (r = 0.363; P = .027). CONCLUSIONS: Hp and 2 of its glycoforms, SAHp and FHp, are potentially useful in the clinical diagnosis of NSCLC. The markers increase with stage, suggesting they may also be useful in stratifying patients at presentation and in following patients after treatment. PMID- 17918262 TI - Metabolic syndrome traits in long-term survivors of pediatric sarcoma. AB - PURPOSE: The metabolic syndrome (MS), a cluster of central obesity, dyslipidemia, hyperglycemia, and hypertension, conveys an increased risk of type 2 diabetes and cardiovascular disease. This cross-sectional study investigated the prevalence of metabolic syndrome traits (MST) in long-term survivors of pediatric sarcoma (SARC) who received multi-modality therapy (MMT). METHODS: Thirty-two SARC survivors (predominantly Ewings; median age 36.5; median age at MMT 15) underwent body composition, activity, and psychosocial analysis. Serum endocrine and inflammatory parameters and urine beta(2)-microglobulin (B2M) were evaluated. The prevalence of MST was compared to age- and gender-matched U.S. population data. RESULTS: SARC survivors were more likely to have two or more MST (OR 2.38 95% CI: [1.14, 5.04]). Analysis of individual MST demonstrated higher prevalence of hypertension (OR 2.61 95% CI: [1.20, 5.59]), hypertriglyceridemia (OR 3.63 95% CI: [1.75, 7.60]), and male visceral abdominal obesity (20-39 years old OR 4.63 95% CI: [0.91, 21.63], 40-59 years old OR infinity). Survivors 18-39 years old had a higher prevalence of the MS (OR 4.29 95% CI: [1.50, 11.21]), defined as three or more MST. Plasminogen activator inhibitory activity (P = 0.016) and B2M (P = 0.027) increased with increasing numbers of MST. In males, total testosterone declined (P = 0.0027) as the number of MST increased. Average (P = 0.014) and maximum (P = 0.021) activity levels decreased as the number of MST increased. CONCLUSION: After a median follow up of 17 years, adult SARC survivors of MMT had an increased prevalence of MST, especially those less than 40 years old. The development of MST in this population was associated with decreased testosterone and activity levels. PMID- 17918263 TI - Lyn tyrosine kinase is required for P2X(4) receptor upregulation and neuropathic pain after peripheral nerve injury. AB - Neuropathic pain, a debilitating chronic pain following nerve damage, is a reflection of the aberrant functioning of a pathologically altered nervous system. One hallmark is abnormal pain hypersensitivity to innocuous stimuli (tactile allodynia), for which effective therapy is lacking, and the underlying mechanisms of which remain to be determined. Here we show that Lyn, a member of the Src family kinases (SFKs), plays an important role in the pathogenesis of neuropathic pain. Nerve injury, but not peripheral inflammation, increased immunoreactivity for active SFKs that were autophosphorylated in the kinase domain (phospho-SFK-IR) in spinal microglia. In spinally derived microglial cells, we identified Lyn as the predominant SFK among the five members (Src, Fyn, Yes, Lck, and Lyn) known to be expressed in the CNS. Lyn expression in the spinal cord was highly restricted to microglia, and its level was increased after nerve injury. We found that mice lacking lyn (lyn(-/-)) exhibit a striking reduction in the levels of phospho-SFK-IR and tactile allodynia after nerve injury, without any change in basal mechanical sensitivity or inflammatory pain. Importantly, lyn(-/-) mice displayed impaired upregulation of the ionotropic ATP receptor subtype P2X(4) receptors (P2X(4)R) in the spinal cord after nerve injury, which is crucial for tactile allodynia. Microglial cells from lyn(-/-) mice showed a deficit in their ability to increase P2X(4)R expression in response to fibronectin, a factor implicated as a microglial P2X(4)R upregulator in allodynia. Together, our findings suggest that Lyn may be a critical kinase mediating nerve injury-induced P2X(4)R upregulation and neuropathic pain. PMID- 17918264 TI - A phase II evaluation of goserelin and bicalutamide in patients with ovarian cancer in second or higher complete clinical disease remission. AB - BACKGROUND: The current study was conducted to determine the effect of goserelin and bicalutamide on progression-free survival (PFS) in patients with epithelial ovarian cancer who were in second or greater complete disease remission. METHODS: Patients received bicalutamide at a dose of 50 mg orally daily and goserelin at a dose of 3.6 mg subcutaneously every 4 weeks. CA 125 was obtained monthly, with computed tomography performed every 3 months. Correlative studies included serum luteinizing hormone, follicle-stimulating hormone, vascular endothelial growth factor, free testosterone, and androstenedione and the germline polymorphisms CYP19A1 and androgen receptor. RESULTS: Between October of 2000 and October of 2002, 35 patients were enrolled. Three patients (9%) received therapy at the time of first disease remission and were removed from the study, and 1 patient (3%) was removed for liver function test abnormalities. The most frequent toxicities were grade 1 alkaline phosphatase (54%), fatigue (57%), and hot flashes (42%) based on the National Cancer Institute common toxicity scale, version 2.0. The PFS for patients receiving protocol therapy in second disease remission (21 patients) was 11.4 months (95% confidence interval [95% CI], 10.2-12.6 months). The PFS for patients receiving protocol therapy in third or fourth disease remission (11 patients) was 11.9 months (95% CI, 10.8-14.1 months). The percentage of patients remaining in second disease remission at given times are: 100% at 3 months, 100% at 6 months, 72% at 9 months, 47% at 12 months, 28% at 15 months, 22% at 18 months, 19% at 21 months, and 13% at 24 months. There were no associations noted between androgen receptor repeat number, genotype, allelotype, or haplotypes and PFS. CONCLUSIONS: The use of goserelin and bicalutamide did not appear to prolong PFS in patients with epithelial ovarian cancer in second or greater complete disease remission. The number of patients in disease remission at given time points may serve as a clinical trial endpoint for future studies of consolidation therapy. PMID- 17918265 TI - Bumetanide enhances phenobarbital efficacy in a neonatal seizure model. AB - OBJECTIVES: High levels of expression of the Na+-K+-2Cl- (NKCC1) cotransporter in immature neurons cause the accumulation of intracellular chloride and, therefore, a depolarized Cl- equilibrium potential (E(Cl)). This results in the outward flux of Cl- through GABA(A) channels, the opposite direction compared with mature neurons, in which GABA(A) receptor activation is inhibitory because Cl- flows into the cell. This outward flow of Cl- in neonatal neurons is excitatory and contributes to a greater seizure propensity and poor electroencephalographic response to GABAergic anticonvulsants such as phenobarbital and benzodiazepines. Blocking the NKCC1 transporter with bumetanide prevents outward Cl- flux and causes a more negative GABA equilibrium potential (E(GABA)) in immature neurons. We therefore tested whether bumetanide enhances the anticonvulsant action of phenobarbital in the neonatal brain METHODS: Recurrent seizures were induced in the intact hippocampal preparation in vitro by continuous 5-hour exposure to low Mg2+ solution. The anticonvulsant efficacy of phenobarbital, bumetanide, and the combination of these drugs was studied RESULTS: Phenobarbital failed to abolish or depress recurrent seizures in 70% of hippocampi. In contrast, phenobarbital in combination with bumetanide abolished seizures in 70% of hippocampi and significantly reduced the frequency, duration, and power of seizures in the remaining 30% INTERPRETATION: Thus, alteration of Cl- transport by bumetanide enables the anticonvulsant action of phenobarbital in immature brain. This is a mechanistic demonstration of rational anticonvulsant polypharmacy. The combination of these agents may comprise an effective therapy for early-life seizures. PMID- 17918266 TI - Frequency, risk factors, and trends for venous thromboembolism among hospitalized cancer patients. AB - BACKGROUND: Venous thromboembolism (VTE) contributes to morbidity and mortality in cancer patients and is a frequent complication of anticancer therapy. In the current study, the frequency, risk factors, and trends associated with VTE were examined among hospitalized cancer patients. METHODS: A retrospective cohort study was conducted using the discharge database of the University HealthSystem Consortium. This included 1,824,316 hospitalizations between 1995 and 2003 at 133 U.S. medical centers. RESULTS: Among 1,015,598 cancer patients, 34,357 (3.4%) were diagnosed with deep venous thrombosis and 11,515 with pulmonary embolism (PE) (1.1%) for an overall VTE rate of 4.1%. Subgroups of cancer patients with the highest rates included black ethnicity (5.1% per hospitalization) and those receiving chemotherapy (4.9%). Sites of cancer with the highest rates of VTE included pancreas (8.1%), kidney (5.6%), ovary (5.6%), lung (5.1%), and stomach (4.9%). Among hematologic malignancies, myeloma (5%), non-Hodgkin lymphoma (4.8%), and Hodgkin disease (4.6%) had the highest rates of VTE. The rate of VTE increased by 28%, secondary to a near-doubling of PE rates from 0.8% to 1.5% (P < .0001). Among patients receiving chemotherapy, the rates of VTE rose from 3.9% to 5.7%, an increase of 47% (P < .0001). In multivariate analysis, risk factors associated with VTE included age >or=65 years, female sex, black ethnicity, use of chemotherapy, primary site of cancer, presence of comorbidities, and year of admission. CONCLUSIONS: VTE, particularly PE, is an increasingly frequent complication of hospitalization in cancer patients. Patients with black ethnicity, specific sites of cancer, or those receiving chemotherapy are disproportionately at risk. PMID- 17918267 TI - FTY720 modulates human oligodendrocyte progenitor process extension and survival. AB - OBJECTIVE: FTY720, a sphingosine-1-phosphate (S1P) receptor agonist that crosses the blood-brain barrier, is a potential immuno-therapy for multiple sclerosis. Our objective was to assess the effect of FTY720 on process extension, differentiation, and survival of human oligodendrocyte progenitor cells (OPCs), and link the functional effects with S1P receptor expression and signaling. METHODS: Functional assays and receptor expression studies were conducted on A2B5+ OPCs derived from the human fetal central nervous system. Cells were treated with physiologically relevant concentrations of the active phosphorylated form of FTY720. S1P receptor/signaling modulators were used to elucidate the basis of the FTY720-induced functional responses. RESULTS: Short-term (1 day) FTY720 treatment caused initial process retraction that was reversed by uncoupling S1P3 and 5 from their G protein using suramin, and with a Rho-kinase inhibitor H1152. Retraction was associated with RhoA-mediated cytoskeletal signaling and with inhibition of OPC differentiation into more mature phenotypes. Continued FTY720 treatment (2 days) induced process extension and enhanced cell survival associated with increased extracellular signal-regulated kinases 1 and 2 phosphorylation, mimicked with the S1P1-specific agonist SEW2871, but not reversed with suramin. Quantitative real-time polymerase chain reaction showed that FTY720 induced reciprocal and cyclic modulation of S1P1 and S1P5 messenger RNA levels. The observed initial downregulation of S1P5 and subsequently of S1P1 messenger RNA supports functional responses being mediated sequentially by S1P5- and later S1P1-associated signaling. INTERPRETATION: FTY720 induces time dependent modulation of S1P receptors on human OPCs with consequent functional responses that are directly relevant for the remyelination process. PMID- 17918269 TI - Unusual nanoparticle structures from the silica sol-gel-mediated self-assembly of a prussian-blue analogue and the formation of templated graphite regions. PMID- 17918268 TI - High-density lipoprotein administration attenuates liver proinflammatory response, restores liver endothelial nitric oxide synthase activity, and lowers portal pressure in cirrhotic rats. AB - In patients with cirrhosis, endotoxic shock is a major complication of portal hypertension, which is related partly to intrahepatic endothelial nitric oxide synthase (eNOS) down-regulation. High-density lipoproteins (HDLs), whose plasma levels are reduced in cirrhosis, have an anti-inflammatory effect by neutralizing circulating lipopolysaccharide (LPS), and they increase eNOS activity in endothelial cells. Therefore, the aim of this study was to assess the effects of reconstituted high-density lipoprotein (rHDL) administration on the LPS-induced proinflammatory response, intrahepatic eNOS regulation, and portal hypertension in cirrhotic rats. Cirrhotic and control rats were pretreated with rHDL or saline and challenged with LPS or saline. The neutralization of LPS in HDL was assessed by the measurement of HDL-bound fluorescent LPS levels. Plasma tumor necrosis factor alpha (TNFalpha) and lipopolysaccharide binding protein (LBP) levels were measured. The expression of hepatic TNFalpha, LBP, inducible nitric oxide synthase (iNOS), and caveolin-1 (a major eNOS inhibitor) and the activity of protein kinase B (Akt; a major eNOS activator) and eNOS were determined. The portal pressure was measured. The plasma HDL levels were significantly lower in cirrhotic rats than in control rats. In cirrhotic rats, the plasma levels of HDL bound fluorescent LPS were 50% lower than those in controls, and they were restored after rHDL administration. The plasma TNFalpha levels were significantly higher in LPS-challenged cirrhotic rats than in controls and significantly decreased after rHDL administration. rHDL administration decreased hepatic TNFalpha, LBP, iNOS, and caveolin-1 expression, restored hepatic eNOS and Akt activity, and significantly lowered the portal pressure and intrahepatic vascular resistance. CONCLUSION: In cirrhotic rats, rHDL administration decreases the hepatic proinflammatory signals induced by LPS, restores the hepatic eNOS activity, and lowers the portal pressure. This suggests that the decrease in circulating HDL in cirrhosis plays a role in the excessive proinflammatory response and intrahepatic eNOS down-regulation. PMID- 17918271 TI - Insight into solid-state entropy from diffraction data. PMID- 17918270 TI - Benzoylurea oligomers: synthetic foldamers that mimic extended alpha helices. PMID- 17918272 TI - Enzyme-responsive materials: chirality to program polymer reactivity. PMID- 17918273 TI - Synthesis and enantioselectivity of p-chiral phosphine ligands with alkynyl groups. PMID- 17918274 TI - Ni-catalyzed cascade formation of C(sp3)--C(sp3) bonds by cyclization and cross coupling reactions of iodoalkanes with alkyl zinc halides. PMID- 17918275 TI - High-activity, single-site mesoporous Pd/Al2O3 catalysts for selective aerobic oxidation of allylic alcohols. PMID- 17918276 TI - Gas-phase infrared spectroscopy and multidimensional quantum calculations of the protonated ammonia dimer N2H7+. PMID- 17918277 TI - Pancreatic scintigraphy with 99mTc-interleukin-2 at diagnosis of type 1 diabetes and after 1 year of nicotinamide therapy. AB - BACKGROUND: To evaluate the clinical utility of pancreatic scintigraphy with 99mTc-interleukin-2 to identify Type 1 diabetic patients with pancreatic inflammation at diagnosis. METHODS: 99mTc-interleukin-2 scintigraphy was performed on 42 newly diagnosed Type 1 diabetic patients, before and after 1 year of treatment with nicotinamide (25 or 50 mg/kg/day) in addition to intensive insulin therapy. Metabolic status was monitored every 3 months for 1 year. Sixteen normal subjects were studied as control. RESULTS: Significant pancreatic accumulation of 99mTc-interleukin-2 was found in 31% of the patients at the time of diagnosis. Patients positive or negative for pancreatic accumulation of interleukin-2 scintigraphy did not show any difference in metabolic or immunologic parameters at diagnosis. Positive patients, however, showed higher C peptide values at 3 months and lower insulin requirement at 1 year, compared to negative patients (insulin requirement (IR): 0.33+/-0.11 vs 0.67+/-0.24 IU/kg/day, positive vs negative patients; p=0.0001); patients positive to IL2 scintigraphy treated with nicotinamide at 25 mg/kg were the only group showing a significant reduction in IR 1 year after diagnosis (IRt0: 0.53+/-0.30 vs IRt12: 0.28+/-0.07 IU/kg/day; p=0.013). After 1 year, all the positive patients showed a significant decrease in pancreatic uptake of 99mTc-interleukin-2 (P/B: 7.87+/ 2.28 at diagnosis vs 5.00+/-1.23 after 1 year; p<0.0001 paired t-test). CONCLUSION: 99mTc-interleukin-2 scintigraphy at diagnosis of Type 1 diabetes may identify patients with pancreatic inflammation. In such patients, treated with nicotinamide at 25 mg/kg, insulin requirement and pancreatic inflammation after 1 year were significantly reduced suggesting that IL2 scintigraphy may be of potential use for assessing the autoimmune phenomena in endocrine pancreas. PMID- 17918278 TI - Sequence and priming in 15 month-olds' reactions to brief arm restraint: evidence for a hierarchy of anger responses. AB - Brief, gentle arm restraint is widely used in experimental studies of children's anger, but the pattern of responses generated by such restraint has been incompletely described. We now describe a hierarchy of responses within trials as well as an escalation across trials that have both methodological and theoretical significance. Mothers of 87 15-month olds prevented them from playing with a toy by restraining their arms on two consecutive 30 sec trials. Physical struggling was the first and most frequent response; children who struggled were significantly more likely to vocalize, and those who vocalized were significantly more likely to show facial expressions of anger. The children's responses became more probable, rapid, and intense during Trial 2 restraint. Overall, the hierarchy was orderly enough to meet criteria for Guttman scalability. The particular sequence observed suggests situational, as opposed to bio-energetic, ordering of responses. Methodologically, the two trial paradigm is a simple, ecologically valid model for studying anger escalation that parallels the "attack priming" of aggression in other species. The magnitude and persistence of anger priming may provide novel measures of anger regulation. Theoretically, the existence of an orderly response hierarchy is consistent with previous observations suggesting that, within a situational context, the sequential appearance of specific behaviors may indicate progressive increases in anger intensity. PMID- 17918279 TI - Weakly aggressive behaviour towards nymphs in the cockroach Schultesia nitor (Blattaria: Zetoborinae). AB - This paper describes aggressive behaviour in the cockroach Schultesia nitor, a tropical forest species living in bird nests. Young S. nitor nymphs are known to show active dispersal while old nymphs and adults are contrastingly strongly gregarious, a combination of features never observed in other cockroach species. Our laboratory experiments using video recording of confrontations between pairs show that aggressive behaviour towards conspecific nymphs is not exhibited towards nymphs of the species Phoetalia pallida, and thus can be considered species specific in S. nitor. But, it is not kin oriented: the mother and all adults of both sexes in different physiological states exhibit this behaviour as well. Six types of aggressive interactions were discriminated, occurring in age symmetric pairs of nymphs and adults. Even more frequent aggression was exhibited by adults and last instar nymphs towards younger nymphs of all instars. The frequency of aggressive acts and types of aggressive interactions varied according to sex and size of the two interacting individuals. The possible function and evolution of this behaviour is discussed, with emphasis on the difficulty of interpreting obvious but weak and not kin-biased aggression. PMID- 17918280 TI - Young adults' media use and attitudes toward interpersonal and institutional forms of aggression. AB - Links between media violence exposure and favorable attitudes toward interpersonal violence are well established, but few studies have examined whether associations extend to include favorable attitudes toward institutional forms of aggression. Studies on this topic have not assessed multiple forms of media use and statistically controlled for individual characteristics likely to influence attitudes beyond sociodemographic information. In this study, undergraduate students (N=319) aged 18-20 years (56% male) completed a survey assessing media use (number of hours per week spent playing videogames, watching movies/TV shows, watching TV sports) and attitudes toward interpersonal violence, punitive criminal justice policies, and different types of military activities (preparedness/defense and aggressive intervention). Greater number of hours spent watching TV contact sports was associated with more favorable attitudes toward military preparedness/defense, aggressive military intervention, and punitive criminal justice policies among men independently of parental education, lifetime violence exposure within the home and community, aggressive personality, and constrained problem solving style. Greater number of hours spent watching violent movies/TV was associated with more favorable attitudes toward military preparedness/defense among men and with more favorable attitudes toward interpersonal violence and punitive criminal justice policies among women, but these associations became non-significant when adjusting for covariates. PMID- 17918281 TI - Women who kill their husbands: mariticides in contemporary Ghana. AB - Homicide by wives against husbands is a largely unexplored subject in lethal violence research. The paucity of information on the phenomenon is particularly acute in the non-Western world where scholarly research is virtually nonexistent. The specific goal for this article, then, was to provide additional insights into the issue by investigating wife-to-husband killings that occurred in Ghana, a non Western society, during 1990-2005. In line with the scant, extant literature, the results of the analysis demonstrate that victims were invariably slain at home. The motive for the crime was to punish a womanizing husband, a husband who had taken another wife, or one who was contemplating wedding another wife in this polygynous society. In other instances, the homicidal intent was to physically eliminate a husband to facilitate an amorous relationship between the assailant and her new lover. In several cases, husband-slayers killed a latent or predisposing victim (e.g. sleeping or ill) via burning, slashing with a machete or food poisoning. The implications of the findings are discussed. PMID- 17918282 TI - FACTS: Fast analytical continuum treatment of solvation. AB - An efficient method for calculating the free energy of solvation of a (macro)molecule embedded in a continuum solvent is presented. It is based on the fully analytical evaluation of the volume and spatial symmetry of the solvent that is displaced from around a solute atom by its neighboring atoms. The two measures of solvent displacement are combined in empirical equations to approximate the atomic (or self) electrostatic solvation energy and the solvent accessible surface area. The former directly yields the effective Born radius, which is used in the generalized Born (GB) formula to calculate the solvent screened electrostatic interaction energy. A comparison with finite-difference Poisson data shows that atomic solvation energies, pair interaction energies, and their sums are evaluated with a precision comparable to the most accurate GB implementations. Furthermore, solvation energies of a large set of protein conformations have an error of only 1.5%. The solvent accessible surface area is used to approximate the nonpolar contribution to solvation. The empirical approach, called FACTS (Fast Analytical Continuum Treatment of Solvation), is only four times slower than using the vacuum energy in molecular dynamics simulations of proteins. Notably, the folded state of structured peptides and proteins is stable at room temperature in 100-ns molecular dynamics simulations using FACTS and the CHARMM force field. PMID- 17918283 TI - Manager-worker-based model for the parallelization of quantum Monte Carlo on heterogeneous and homogeneous networks. AB - A manager-worker-based parallelization algorithm for Quantum Monte Carlo (QMC-MW) is presented and compared with the pure iterative parallelization algorithm, which is in common use. The new manager-worker algorithm performs automatic load balancing, allowing it to perform near the theoretical maximal speed even on heterogeneous parallel computers. Furthermore, the new algorithm performs as well as the pure iterative algorithm on homogeneous parallel computers. When combined with the dynamic distributable decorrelation algorithm (DDDA) [Feldmann et al., J Comput Chem 28, 2309 (2007)], the new manager-worker algorithm allows QMC calculations to be terminated at a prespecified level of convergence rather than upon a prespecified number of steps (the common practice). This allows a guaranteed level of precision at the least cost. Additionally, we show (by both analytic derivation and experimental verification) that standard QMC implementations are not "perfectly parallel" as is often claimed. PMID- 17918285 TI - Melanoma of the head and neck: a review. AB - Melanoma of the head and neck and its treatment are complex issues. The behavior of head and neck melanoma is aggressive, and it has an overall poorer prognosis than that of other skin sites. Current understanding of the behavior of head and neck melanoma is reviewed and treatment strategies are presented. Controversies in treatment include the role of lymphoscintigraphy with sentinel node biopsy, nodal dissection, margin size, role of radiation therapy, and reconstruction. The therapeutic goal is to treat melanoma aggressively while minimizing the effects of treatment on patient's quality of life. Due to its biological behavior, head and neck melanoma should be treated in an aggressive manner when morbidity is not significantly increased. Patient's specific treatment is imperative. PMID- 17918284 TI - Integration of novel targeted therapies into the systemic treatment of breast cancer--a review. AB - Over the last few years it has been anticipated that molecularly targeted agents can provide substantial improvement in the treatment of breast cancer. The most illustrative paradigm has been that of trastuzumab (Herceptin), a humanized monoclonal antibody against the HER2 oncoprotein overexpressed in 25% of breast cancers. Trastuzumab when combined with standard cytotoxic chemotherapy improved the outcome and survival in patients with metastatic breast cancer, whereas, over the last 2 years studies incorporating trastuzumab in sequence to or concurrently with taxane-based chemotherapy in the adjuvant setting demonstrated a considerable benefi t in this subset, with the results of longer follow-up regarding long-term outcome and late toxicities expected over the forthcoming years. Moreover, the prognostic and predictive value of topoisomerase IIa (Topo IIa) overexpression in these subgroups with respect to anthracycline treatment has been extensively discussed and analysed. Other inhibitors of both HER1/HER2 have recently been introduced with promising results and results of ongoing studies are awaited with great interest. A recently anticipated target in advanced breast cancer has been the pathway of angiogenesis; first a humanized monoclonal antibody-bevacizumab (Avastin)- has demonstrated encouraging results when combined with chemotherapy in pretreated HER2-negative advanced breast cancer, while combinations with trastuzumab+/-chemotherapy are currently examined in HER2-overexpressing breast cancer. Furthermore, as novel molecular pathways relevant to breast cancer biology are explored, it is expected that a whole array of targeted agents will be tested in combination or in sequence to standard chemotherapy with the aim to improve outcome of high-risk breast cancer patients. PMID- 17918286 TI - Multimodality treatment of germ cell cancer patients who had progression or relapse after high dose chemotherapy and autologous bone marrow transplantation. AB - PURPOSE: When a patient with germ cell tumor (GCT) fails to be cured with high dose chemotherapy (HDC) and autologous haematopoietic stem cell transplantation (auto- BMT) the overall prognosis is very poor and any further treatment has only palliative character. A question requiring answer is how intense should this kind of treatment be, and what can be expected from it. PATIENTS AND METHODS: Of 44 patients with GCT who were transplanted after HDC in our centre between 1999- 2005, 17 experienced treatment failure. Amongst them 14 had marker-positive relapse or confirmed germ cell histology. Another 3 had second primary neoplasms. Of the 17 patients 14 received further treatment that consisted of surgery alone in 2, chemotherapy in 2, radiotherapy in 1, combined surgery + chemotherapy in 5, chemotherapy +surgery + radiotherapy in 3 and chemotherapy + radiotherapy in 1 patient. RESULTS: The median survival from the time of relapse was 3 months in all patients, and 6 months in the 14 patients who received further treatment. In 6 patients with relapse confined to a single site the median survival was 11 months. Three patients in this group are alive with overall survival (OS) of 37.4+, 24.3+ and 6.2+ months (all had multimodal treatment: chemotherapy + surgery or radiotherapy, and all achieved durable complete response/CR). CONCLUSION: Our results suggest that GCT patients who have relapsed/ progressed after HDC may benefit from further treatment. Best chances for long term survival have those who experience relapse confined to one metastatic site and receive combined treatment (surgery or radiotherapy plus systemic therapy). PMID- 17918287 TI - Neoadjuvant chemotherapy followed by radiotherapy versus concurrent chemoradiotherapy for larynx preservation in patients with advanced laryngeal cancer. AB - PURPOSE: The efficacy and toxicity of neoadjuvant chemotherapy followed by radiotherapy and concurrent chemoradiotherapy and their impact on larynx preservation have been studied in patients with advanced (stage III, IVa, and IVb) squamous cell cancer of the larynx. PATIENTS AND METHODS: Fifty patients were treated with either 2-4 cycles of induction chemotherapy with cisplatin 100 mg/m(2), day 1 and infusional 5-fluorouracil (5-FU 1000 mg/m(2), days 1-5), followed by radiotherapy 70 Gy, 1.8-2 Gy per fraction, or concurrent chemoradiotherapy (the above-mentioned radiotherapy concurrently with carboplatin 300 mg/m(2) every 21 days or weekly paclitaxel 80 mg/m(2)). Patients were allocated in the 2 arms by 1:1 selection. At the end of both protocols, patients without complete response (CR) underwent laryngectomy and/or neck lymph node dissection. Assessed were response and toxicity rates, overall survival (OS) and disease-free survival (DFS). RESULTS: A total of 31 (62%) patients achieved larynx preservation with acceptable organ function. No statistically significant difference in response rate and OS was found between the two treatment arms. Patients submitted to concurrent chemoradiotherapy showed significantly longer DFS (14 vs. 10 months, p= 0.0397) and higher rates of larynx preservation (p <0.05). All grade IV side effects occurred in the concurrent chemoradiotherapy group. CONCLUSION: Concurrent compared to alternating chemoradiotherapy was more toxic, but achieved significantly longer DFS and higher rate of larynx preservation. PMID- 17918289 TI - The impact of treatment interruptions on locoregional control during postoperative breast irradiation. AB - PURPOSE: Patients treated in the years 1990-2003 with breast conserving surgery and postoperative adjuvant radiotherapy were retrospectively analyzed in order to determine the impact of radiotherapy interruptions on the treatment outcome. PATIENTS AND METHODS: 470 patients received breast irradiation with (60)Co or with 4MV photon linear accelerator (total dose 50 Gy, range 46-54). Four hundred and sixty-two (98%) patients had boost dose of 10-20 Gy to the tumor bed either with photon or electron irradiation. Irradiation of the lymphatic fields was carried out according to the number of involved nodes. Even one day of treatment break was accepted as treatment interruption. Unplanned treatment interruptions occurred in 423 (90%) patients. A total of 196 (41%) patients had no treatment breaks or had interruptions of 7 days or less and 274 (59%) patients had treatment interruptions longer than 7 days. Locoregional control (LC) and overall survival (OS) rates were estimated by Kaplan-Meier method. The groups were compared with log-rank x(2) and Pearson x(2) tests. RESULTS: For all patients 5 year LC and OS rates were 91% and 85%, and 10-year rates were 86% and 75%, respectively. LC rates for the group of patients without interruptions or with an interruption of less than a week were 95% for 5 years and 90% for 10 years. Five- and 10-year LC rates for the patients with interruptions of 8 days or more were 87% and 83%, respectively (p=0.02). CONCLUSION: This retrospective study shows that postoperative irradiation interruptions of more than a week negatively impact LC. PMID- 17918290 TI - The effect of Nd:YAG laser resection on symptom control, time to progression and survival in lung cancer patients. AB - PURPOSE: The aim of this study was to determine the effect of Nd:YAG laser resection of centrally located tumors on the control of various symptoms and signs, time to progression and survival in lung cancer patients. PATIENTS AND METHODS: We evaluated the effects of Nd: YAG laser resection in combination with high-dose rate (HDR) brachytherapy and external beam radiotherapy (EBRT) vs. combination of HDR brachytherapy and EBRT alone on lung cancer symptoms and signs, ECOG performance status, time to progression and overall survival in lung cancer patients. Patients in group I (n=81) were treated with combination of HDR brachytherapy and EBRT, while patients in group II (n=97) were treated with Nd:YAG laser in combination with HDR brachytherapy and EBRT. Patients were evaluated before and after treatment, and were followed-up regularly every 3 months until the end of life. After RT +/- laser treatment all patients received standard chemotherapy (cisplatin plus etoposide) during the course of disease. RESULTS: After treatment in both groups significant improvement in all investigated parameters was seen. Improvement in dyspnoea, thoracic pain, body weight loss and ECOG performance status was significantly better in group II (p <0.05), as were time to progression and overall survival (p <0.05). CONCLUSION: Laser resection improves symptom control in lung cancer patients with central airway obstruction (CAO). Longer time to progression and survival of lung cancer patients could be the result of imminent airway desobstruction accomplished with Nd:YAG laser. PMID- 17918288 TI - Myocardial function in patients with breast cancer after therapy with anthracyclines and chest irradiation. AB - PURPOSE: To evaluate and compare the left ventricular performance in female patients with breast cancer treated with chemotherapy (CT) and either left or right thoracic irradiation (RT), using Doppler echocardiography. PATIENTS AND METHODS: Thirty-four patients with cancer of the left breast treated with surgical resection, adjuvant EC CT (epirubicin and cyclophosphamide) and conventionally fractionated left thoracic RT (study group) and a gender-and age matched group of 34 patients with cancer of the right breast also treated with surgical resection, adjuvant EC CT and right thoracic RT (control group), were studied by echocardiography. Assessed were the left ventricular systolic performance by measuring the global ejection fraction (EF) and the shortening fraction (SF). Left ventricular diastolic performance was assessed by measuring the Doppler transmitral flow: the maximal velocity of the E wave (rapid filling/ Emax) and A wave (atrial filling/Emax) were measured. The ratio of Emax/Amax, the pressure half-time (PHT) of the E wave and the isovolumic relaxation time (IVRT) were also calculated. RESULTS: The left ventricular diastolic performance was altered in the study group which showed a significant decrease (p <0.001) of Emax. A wave was significantly increased in the study group compared with the control group (p <0.001. The mitral E/A ratio was subunitary in both groups but more depressed in the study group. The E wave PHT was more prolonged in the study group compared to the control group (p <0.001). The IVRT was prolonged in the study group compared with the controls (p <0.05). The left ventricular systolic performance was within normal limits in both groups. CONCLUSION: Our Doppler echocardiography study documented an impaired left ventricular diastolic performance in patients with cancer of the left breast treated with fractionated thoracic RT and CT. This impairment is due to poor left ventricular compliance. PMID- 17918291 TI - Investigation of circumferential resection margin in rectal cancer surgery. AB - PURPOSE: Literature data suggest that investigation of the circumferential resection margin (CRM) in rectal cancer surgery gives significant information for prognosis of the disease and the quality of the performed operation. The aim of this study was to analyze the clinical and morphological aspects of CRM in pT3 tumors in relation to their significance for rectal cancer treatment and staging. PATIENTS AND METHODS: Thirty patients with radically operated pT3 stage rectal cancer were investigated. The resected specimens were studied both macroscopically and microscopically to assess CRM and the quality of the performed operation. RESULTS: Six (20%) patients had positive CRM (CRM +; tumor cells in the margin or < 1 mm from it). These cases were characterized with deep infiltration (> 4 mm) in the rectal adventitia (perirectal tissue). Statistically significant correlation was found between CRM status and quality of the resected specimen (including the presence of rectal fascia in CRM). There was no correlation between pathological characteristics of the tumor and the status of CRM. CONCLUSION: The status of CRM is a factor independent from TNM classification. The presence of rectal fascia in CRM confirms the good quality of the operation. CRM+ could be a result of inadequate surgery or advanced disease. The status of CRM could be used in the decision-making for postoperative adjuvant treatment. PMID- 17918293 TI - Clinical burden of terminal phase in cancer patients. AB - PURPOSE: Symptoms, palliative applications and terminal phase (TP) duration mainly constitute clinical burden of TP in cancer. The aim of this study was to describe palliative issues and determine factors that might influence the clinical burden during TP. PATIENTS AND METHODS: We retrospectively studied 125 terminally ill cancer patients, of median age 70 years and ECOG performance status (PS) 4, at their TP entry, who died over a 3-year period, 90 (72%) in hospital (group 1) and 35 at home (group 2). RESULTS: During TP, the 10 most prevalent symptoms were anorexia, pain, dyspnoea, cachexia, fever (infectious or not), constipation, ascites, agitation and vomiting. The 10 most frequent palliative applications were administration of i.v. fluids/drugs, oxygen and enemas administration, bladder catheter and central venous (CV) line care, transfusions, gastric, pleural and peritoneal fluid drainage, endoscopic or percutaneous biliary tract decompression, and radiotherapy. In total, TP represented 8% of the whole disease duration, causing 17% of admissions and 28% of hospital stay. Tumor complications and treatment toxicities caused death and shortened TP in 70% of the cases, more frequently in group 1 (p=0.027). Long disease active phase (AP) duration (from initial diagnosis of cancer to TP entry) predicted PS<4 at TP entry (p=0.008). Predictors of TP duration were AP duration (p=0.010), PS <4 (p=0.000) and number of clinical features (p=0.012). CONCLUSION: TP represents a small percentage of patients' total disease duration, but it constitutes a considerable part of the entire hospital patients' in-stay and professional practice. PS at TP entry predicted TP duration. AP influenced PS at TP entry and predicted TP duration. AP is an easily measurable clinical parameter and may be considered as an additional prognostic factor of TP duration. PMID- 17918292 TI - Overall survival in carcinoma of the penis--a single institution study. AB - PURPOSE: This retrospective study was performed to evaluate the overall survival (OS) and to identify some prognostic factors and their influence on the survival of patients with squamous cell carcinoma of the penis (SCCP). PATIENTS AND METHODS: Fifty-nine patients with SCCP, treated at the Clinic of Urology, Medical University, Plovdiv, between 1980 and 2005, were studied. The patients' OS was estimated by the Kaplan-Meier life table method. Log-rank test was used to compare cumulative survival functions between different groups of patients. RESULTS: During a mean follow-up period of 7.06+/-0.97 years (range 2 months-26 years; median 4 years) 37 (62.71%) patients died (SCCP progression 29, intercurrent illness 7, other cause 1), giving a mean survival time of 12.56 years )95% confidence interval [CI] = 9.46-15.67; median 14 years). The 5-year OS was 62.19%. The main adverse prognostic factors significantly affecting the OS were: age (p= 0.0199), pathological (p) T stage (p= 0.0001), pN status (p= 0.0046) and pN stage (p= 0.0001). CONCLUSION: The prognostic factors identified in this study are of great importance for the clinical practice, especially for decision making over the treatment strategy involving primary tumor and regional lymph nodes. PMID- 17918294 TI - Quality of life and treatment related toxicity in 335 patients with locally advanced cervical carcinoma treated by two chemoradiation regimens. AB - PURPOSE: Concomitant chemoradiotherapy (CT/RT) is the gold standard for advanced cervical carcinoma, but with frequent debates over treatment schedules and toxicity. This study compared 2 concomitant CT/RT regimens in terms of quality of life (QoL) and acute toxicity. PATIENTS AND METHODS: Between March 2003 and March 2005, 335 patients with stage IIB-IIIB cervical carcinoma were evaluated in a randomized single-center phase III trial at the Oncology Institute Cluj-Napoca. Patients received concurrent CT/RT with cisplatin 20 mg/m(2), days 1-5, every 21 days (arm A, n=171) or 40 mg/m(2)/weekly (arm B, n=164). QoL was estimated using the EORTC QLQ-30, v.3.0 questionnaire and acute toxicity using the common toxicity criteria (CTC) v.2.0. RESULTS: Significant improvement of global health status (p <0.01) and a decrease in pain (p <0.01) was observed in arm A. In arm B fatigue increased (p=0.01) and role functioning diminished (p=0.05). In both arms depression, nausea, vomiting and diarrhea increased (p <0.05). Gastrointestinal toxicity was similar in both arms (76% vs. 77.5%). Hemoglobin drop was higher in arm B: 75% vs. 63% (p=0.02), while no differences were seen in leukocyte and platelet toxicity. CONCLUSION: Concomitant CT/RT with cisplatin 20 mg/m(2) x 5 days every 21 days has better impact on patients' QoL and lower toxicity compared with the weekly chemotherapy regimen. PMID- 17918295 TI - Assessment of the quality of life in patients with oral cavity and jaw cancer in three regions of Bulgaria. AB - PURPOSE: The aim of the present study was to assess the quality of life (QoL) in 103 patients with oral cavity and jaw cancer, using an original rating scale. PATIENTS AND METHODS: In order to assess the QoL in such patients an original rating scale was developed and validated. For statistics alternative analysis, non parametric tests, correlation analysis, and graphic analysis were used. RESULTS: Most patients were rated as grade III or IV according to the 5-grade scale (grade IV the most severe). Their QoL was significantly deteriorated, resulting in depression and despair. Reconstruction and prosthesis implantation and social rehabilitation were needed in these patients. QoL was assessed in 19 patients prior to and following prosthesis implantation. Prior to implantation most patients were rated as grade III or IV, whereas the majority of them were rated as grade 0 or 1 following prosthesis implantation. Prosthesis implantation contributed to improvement of speaking and feeding and elimination of disfigurement. CONCLUSION: After assessment of QoL, patients rated as grade III or IV should have access to complex medical management, including prosthesis implantation and social rehabilitation. PMID- 17918297 TI - Secondary acute myeloid leukemia early after therapy for Hodgkin's disease--a case report. AB - A case of acute myeloid leukemia (AML) after successful therapy for Hodgkin's disease (HD) is reported. The patient was diagnosed with stage IIB HD at the age of 25. She was treated with chemotherapy and radiotherapy (RT), and complete remission (CR) was achieved. Seven months after the CR was obtained the patient developed AML. Knowing that the prognosis of patients with secondary AML (sAML) after primary HD is poor we decided to perform autologous peripheral stem cells' transplantation. PMID- 17918296 TI - Synchronous bilateral seminoma: report of a case and literature review. AB - Bilateral occurrence of a primary testicular tumor is unusual. The reported incidence of bilateral germ cell tumors of the testes has been between 0.5 and 7% and the majority of these cancers (90%) are metachronous. We report the case of a 27-year-old man with synchronous bilateral seminoma and discuss the value of the histopathological diagnosis in therapy and prognosis. The related literature is concisely reviewed. PMID- 17918298 TI - Non specific magnetic resonance features of chondromyxoid fibroma of the iliac bone. AB - Chondromyxoid fibroma is a benign cartilaginous neoplasm. The tumor is rare, perhaps the rarest of all bone tumors. It is more common in long bones and rare in pelvic bones. We report a case of a 31-year-old woman with chondromyxoid fibroma arising from the iliac bone and presenting with leg pain due to compression on the femoral nerve. Magnetic resonance imaging (MRI) features were non specific for chondromyxoid fibroma. We emphasize the need to consider it in the differential diagnosis of chondrosarcoma. PMID- 17918299 TI - Endocrine paraneoplastic syndromes in small cell lung carcinoma. Two case reports. AB - Paraneoplastic syndromes are frequently detected in many small cell lung cancer (SCLC) patients. In the present paper we report 2 cases of patients diagnosed with SCLC, in whom 2 distinct endocrine paraneoplastic syndromes were identified during diagnosis. In the first patient, severe hyponatremia and renal sodium loss with inappropriate antidiuresis was found during routine laboratory tests. Serum antidiuretic hormone (ADH) level was within normal limits, but the atrial natriuretic peptide (ANP) level was elevated. The second case presented with severe hypercalcemia secondary to an excessive parathormone (PTH) secretion. We discuss the 2 cases and review the literature. PMID- 17918300 TI - What does impact factor depend upon? AB - Nobody doubts the importance of the scientific performance's evaluation. The journal impact factor is increasingly employed to evaluate the quality of scientific research. The use of term "impact factor" has gradually evolved, especially in Europe, to include both journal and author impact. This ambiguity often causes problems. It is one thing to use impact factors to compare journals and quite another to use them to compare authors. Journals impact factors generally involve relatively large numbers of articles and citations. Individual authors, on average, produce much smaller numbers of articles. Many scientists consider that impact factor is not the perfect tool to measure the quality of articles but there is nothing better and it has the advantage of already being in existence and is, therefore, a good technique for scientific evaluation. However, the use of journal impact factor is probably the most controversial issue. PMID- 17918301 TI - Haemagiopericytoma: a rare tumor with an eventual malignant clinical behavior. PMID- 17918302 TI - John Hunter (1728-1793): founder of scientific surgery and precursor of oncology. AB - In the following article we briefly present the life and scientific work of John Hunter, founder of the experimental surgery, who by means of his researches and writings in the field of Oncology, contributed enormously to the development of this discipline. In reference to this matter, we quote some examples indicative of his work on Oncology. PMID- 17918303 TI - General practice consultations. PMID- 17918304 TI - Early detection of prostate cancer. PMID- 17918305 TI - Vitamin D deficiency. PMID- 17918306 TI - What motivates physicians throughout their careers in medicine? AB - Motivation provides direction and purpose in physicians' work, and motivating factors vary during different career stages. Motivation theories divide sources of motivation into those intrinsic to the work, such as the opportunity for self expression and intellectual challenge, and those extrinsic to the work, such as salary and time. Although much attention has focused on minimizing negative extrinsic factors, the authors argue that career resilience requires that physicians reflect on and define the sources of their own intrinsic motivation. Opportunities to maximize self-awareness may allow physicians to structure their work in ways that maximize meaning and fulfillment over the long-term. PMID- 17918307 TI - [A 33-year old pregnant woman with nausea, itching and fatigue]. AB - Acute fatty liver of pregnancy (AFLP) is a serious, but rare condition with substantial maternal and perinatal mortality and morbidity. It occurs in the third trimester or early postpartum period. The medical history, physical examination and laboratory tests are often sufficient to make the diagnosis, and liver biopsy is rarely indicated. We present a 33-year-old woman with the diagnosis AFLP. Her case is presented to draw attention to AFLP as a differential diagnosis to liver diseases in pregnancy, especially the HELLP syndrome (haemolysis, elevated liver enzymes and low platelets). Appropriate diagnosis and prompt delivery is essential to optimal maternal and fetal outcome in both the AFLP and HELLP syndromes, and this should be followed by intensive care treatment of the dysfunctional maternal multiorgan system. PMID- 17918308 TI - [Itching in pregnancy is mostly--but not always--harmless]. PMID- 17918310 TI - [Non-physician clinicians common in Africa]. PMID- 17918309 TI - GA perception and signal transduction: molecular interactions of the GA receptor GID1 with GA and the DELLA protein SLR1 in rice. PMID- 17918311 TI - [Chondroma of the subvocal part of the larynx]. PMID- 17918312 TI - More on thunderstorms and iPods. PMID- 17918313 TI - Re: Patient privacy and Latin: my father's story. PMID- 17918314 TI - Teen unsafe driving behaviors: focus group final report. PMID- 17918315 TI - Re: Patient privacy and Latin: my father's story. PMID- 17918316 TI - Survival pathways in hypertrophy and heart failure: the gp130-STAT axis. AB - Circulating levels of interleukin (IL)-6 and related cytokines are elevated in patients with congestive heart failure and after myocardial infarction. Serum IL 6 concentrations are related to decreasing functional status of these patients and provide important prognostic information. Moreover, in the failing human heart, multiple components of the IL-6- glycoprotein (gp)130 receptor system are impaired, implicating an important role of this system in cardiac pathophysiology. Experimental studies have shown that the common receptor subunit of IL-6 cytokines is phosphorylated in response to pressure overload and myocardial infarction and that it subsequently activates at least three different downstream signaling pathways, the signal transducers and activators of transcription 1 and 3 (STAT1/3), the Src-homology tyrosine phosphatase 2 (SHP2) Ras-ERK, and the PI3K-Akt system. Gp130 receptor mediated signaling promotes cardiomyocyte survival, induces hypertrophy, modulates cardiac extracellular matrix and cardiac function. In this regard, the gp130 receptor system and its main downstream mediator STAT3 play a key role in cardioprotection. This review summarizes the current knowledge of IL-6 cytokines, gp130 receptor and STAT3 signaling in the heart exposed to physiological (aging, pregnancy) and pathophysiological stress (ischemia, pressure overload, inflammation and cardiotoxic agents) with a special focus on the potential role of individual IL-6 cytokines. PMID- 17918317 TI - [Agreed terminology--a criterion of objective assessment of the results of extracorporeal shock-wave lithotripsy]. AB - The decision on administration of extracorporeal shock-wave lithotripsy for uroliths should be based on the clinical picture, physicochemical properties of the urolith and anatomofunctional state of the kidneys and upper urinary tracts. Information about the treatment conducted should be maximally complete and significant as this allows objective interpretation of the treatment results and design of further treatment. Usage of agreed terminology facilitates the analysis of the protocols, guidelines and assessment of treatment results in clinic and prophylactic centers. PMID- 17918318 TI - [Diagnosis, prevention and treatment of renal lesion in extracorporeal shock wave lithotripsy]. AB - The diagnosis and treatment of 120 patients with urolithiasis were made. The following examinations were made: the study of immune status and lipid peroxidation, detection of selective proteinuria, leukocyte migration inhibition reaction with autoantigens obtained from the tissues of the renopelvic segment providing additional information about renal parenchyma and immune system. The patients were divided into two groups: 70 patients of the study group received pre-, intra- and postoperative preventive pharmacotherapy, while 50 patients of the control group received standard therapy. It is shown that preventive measures in the patients of the study group aimed at raising resistance of the kidney to the shock wave and prevention of infectious-inflammatory process evidence for necessity of the above measures not only before extracorporeal shock wave lithotripsy, but also in intra- and postoperative periods. PMID- 17918319 TI - [Paraurethral cyst and/or urethral diverticulum--differential diagnosis]. AB - From 2003 to 2006 we examined and treated 73 females with paraurethral lesions. The differential diagnosis was made using gynecological and US examinations, urethral profilometry, miction cystourethrography, high-pressure urethrography, MR imaging. All the patients have undergone extirpation of paraurethral cyst/diverticulectomy. The diagnosis of paraurethral lesions should be made with application of all available methods. Gynecological examination is the first procedure in complex diagnostic investigation, ultrasound must be also used in all the patients with addition of x-ray in case of insufficient data. MR imaging is used in large size of paraurethral lesions, their connection with urethra. Treatment of such patients is surgical. PMID- 17918320 TI - [Videoendoscopic ligation of the internal seminal veins in bilateral varicocele]. AB - Varicocele diagnosis should be based not only on physical methods but on ultrasound dopplerography as well (flebography or scrotal thermography) to detect a subclinical form and bilateral process. Bilateral varicocele (BV) occurs more often than is diagnosed. It results in more changes in spermogram and is a prognostic infertility factor. Bilateral pain, impairment of spermatogenesis and infertility are indications for bilateral ligation of the seminal veins in BV patients. Bilateral ligation is much more effective than unilateral. Laparoscopic and microsurgical methods are most effective and low invasive in correction of BV and present a method of choice in the treatment of such patients. PMID- 17918322 TI - [Thickening phalloplasty with application of allogenic transplant]. AB - Sixteen patients have undergone thickening phalloplasty with allogenic transplant alloplant. As the result of phalloplasty penis thickness increased by 16.7%. Fifteen (93.8%) patients were satisfied with a new size of the penis, one patient (6.3%) was disappointed. Three patients had infectious complications. The transplant showed excellent engraftment. Thus, allogenic transplant alloplant is an appropriate biomaterial for thickening phalloplasty. PMID- 17918323 TI - [Efficiency of clinical examination of patients with sarcoidosis under the conditions of general health care network]. AB - The efficiency of clinical examination of patients with sarcoidosis was compared in the general health care network and tuberculosis facilities. The follow-up of patients with respiratory sarcoidosis in the tuberculosis facility in 1996-2002 was found to be better: centralized monitoring of the prevalence and incidence of sarcoidosis was exercised, the time of diagnostic search and the number of diagnostic errors substantially reduced, control examinations were performed to early detect a relapse and to reduce disability rates among patients with sarcoidosis. It is suggested that it is expedient to resume a follow-up of patients with sarcoidosis in a special room of a polyclinic or a tuberculosis dispensary. PMID- 17918321 TI - [Treatment of erectile dysfunction in patients with ischemic heart disease and angina of effort]. AB - A total of 58 patients with ischemic heart disease and angina of effort (FC II) (mean age 55.6 years) participating in the study were diagnosed to have erective dysfunction (ED) of a mild (35%), moderate (57%) and severe degree (8%). All the patients were randomized into two groups. Group 1 (n=21, mean age 56.4 years) received standard cardiotropic therapy (nitrates, beta-blockers, ACE inhibitors on demand, diuretics, antioxidants) and placebo. Group 2 (n=37, mean age 54.3 years) received the same standard cardiotropic therapy plus impase (1 tablet each other day for 3 months). The results of the trial show that impase addition to cardiotropic therapy raised exercise tolerance, diminished the number of anginal attacks in mild and moderate exercise, enhanced coronary microcirculatory blood flow, increased reserve circulation index by 34%, improved metabolism of vascular endothelium in the whole body. Impase acts pathogenetically in endothelial insufficiency. Prevention of endothelial dysfunction by impase allows both to stop progression of cardiovascular disease and to prevent erectile dysfunction. PMID- 17918324 TI - [Work quota setting in a phthisiatrician during outpatient reception hours (as a consulting and local physician)]. AB - The study was undertaken to determine a phthisiatrician's man-hours during outpatient hours as a consulting and local physician in the Stavropol Territorial Tuberculosis dispensary. It used an analytical method - man-hours for individual components of work, i.e. working operations at photochronometric monitoring. The latter of the work of two consulting physicians and two local ones (two working weeks per specialist) was made. The 2006 phthisiological care schedule envisaged 185.6 visits per 1,000 dwellers. If these data are taken as the basis, then 0.64 of the position of a phthisiatrician per 10,000 is required to deliver health care to patients with tuberculosis, by taking into account the work quota setting developed in this study. An expert evaluation was to substantiate the labor hours of a phthisiatrician when he delivered outpatient health care to patients with tuberculosis. Expertise of the substantiation of the labor hours of a phthisiatrician delivering outpatient health care to patients with tuberculosis has revealed that the maximum working time spent by the phthisiatrician on medical documentation work, to the prejudice of the quality of patient examination, the raising the level of his skills, and the performance of preventive work among the patients. PMID- 17918326 TI - [Comparative analysis of tuberculosis morbidity trends in the penitentiary and civil health care systems of the Udmurt Republic over 10 years]. AB - Pulmonary tuberculosis morbidity trends were comparatively analyzed in the penitentiary and civil health care systems of the Udmurt Republic (UR) over 10 years (1996-2005). Two periods were identified in the course of a tuberculous process. In the first period (1996-2000), the increase in pulmonary tuberculosis morbidity rates in the UR penitentiary system was 3 times greater than the republican rates, the rise in this index in the UR penitentiary system being 25.3 times higher than that in the civil health system. In the second period (2001 2005), the UR penitentiary system was marked by a good tendency towards the stabilization of tuberculosis morbidity rates. At the same time, in the UR civil health care system, there is a further increase in tuberculosis morbidity rates in the population, although it occurs more slowly. PMID- 17918325 TI - [The clinical and epidemic characteristics of new-onset tuberculosis in relation to the procedure of detection]. AB - The outpatient cards and case histories of 456 patients with tuberculosis detected in Samara in the period from January 1, 1999 to December 31, 2003 were examined. Tuberculosis was identified in 58.3% of the patients when they were asking for medical aid; among them 54.2% were treated at general hospital where caseous pneumonia, disseminated tuberculosis, and tuberculous pleurisy were more frequently diagnosed. The unemployed able-bodied persons amounted to 43% of the detected patients with tuberculosis, of whom 81% being identified on recourse to a doctor. Primary drug resistance does not depend on the procedure of tuberculosis detection. Early detection and the initiation of treatment will improve a clinical prognosis in patients with new-onset tuberculosis. PMID- 17918327 TI - [Treatment policy for craniovertebral spondylitis]. AB - The paper summarizes the results of surgical treatment in 25 patients with atlantoaxial spondylitis, including 15 with spondylitis of tuberculous etiology and 10 with nonspecific etiology. Ten patients were found to have severe neurological disorder requiring application of skeletal traction behind the malar arches in 5 cases and that of a halo-apparatus to 2 cases in order to eliminate the occurred dislocation and to relieve neurological symptoms. According to the P. Denis concept adapted for the craniovertebral region, all the patients were divided into 3 groups in relation to the degree of anatomic and biomechanical changes. Group 1 patients received medical treatment with firm collar fixation. Group 2 patients with anterior pillar injury underwent transpharyngeal sanitation of an infectious focus and occipitospondylodesis. Decompressive stabilizing operations were performed in all the patients, which consisted of two stages: 1) occipitospondylodesis made, by using a titanium wire and No. 91/198 Simplex bone cement (Howmedica, United Kingdom); 2) transoral access with abscess cavity sanitation; removal of necrotic tissues; thrifty resections of bone tissue portions within the intact tissues; anterior stabilization of the affected portion with an autologous bone graft. Postoperatively, the cervical spine was fixed with a head holder for as long as 3-5 months (until there were clear X-ray signs of bony ankylosis (block) in the area of osteoplasty). Quaternary antituberculous therapy was compulsorily made for 6-9 months. The described procedure for surgical treatment of craniovertebral spondylitis could yield positive results in all cases, by abolishing neurological symptoms and forming bony ankylosis in the atlantoaxial area to restore the supporting capacity of the cervical spine. PMID- 17918328 TI - [Impact of endogenous porphyrin-photosensitized exposure on peripheral lymphocytes in patients with pulmonary tuberculosis]. AB - Peripheral lymphocytes (PL) were studied in 63 patients with pulmonary tuberculosis and 28 donors. The study revealed the elevated levels of lipid peroxidation products, altered membrane potential, and diminished PL viability in patients with pulmonary tuberculosis as compared with healthy donors. Analysis of the time dependences of cell viability and production of active tuberculosis products after endogenous porphyrin-photosensitized exposure suggests that the lymphocytes from patients with pulmonary tuberculosis are more exposed to the damaging effect of free radicals than the cells from healthy individuals. This is more apparent 18 hours of dark incubation after cell illumination in the presence of ALK-induced porphyrins in doses of 47.4 kJ/m2 or more. The findings suggest that the changes occurring in PL from patients with pulmonary tuberculosis may substantially cause immune disorders and contribute to the progression of a tuberculous process. PMID- 17918329 TI - [Academician S. N. Fyodorov's contribution to Russian and world ophthalmology]. AB - The article is dedicated to academician S. N. Fyodorov, an outstanding surgeon, scientist, and a public figure. The paper covers the milestones of S. N. Fyodorov's creative way. His motto was uncompromisingness, diligence, and innovation. The main result of academician Fyodorov's many-year work, his brainchild, was Eye Microsurgery Complex, an inter-branch scientific and technical establishment, which became the material base for his most daring projects. S. N. Fyodorov made a real revolution in ophthalmology by changing it from a modest surgeless science into a bright and stormily progressing field of medicine. Thanks to his achievement, Russia is still one of the world's leaders in ophthalmology. Today, Eye Microsurgery is the flag of Russian ophthalmology. It provides more than 50% of all high-technology ophthalmologic aid in the country. PMID- 17918330 TI - [The national "Health" project and its implementation in the Penza region]. PMID- 17918331 TI - [The estimated glomerular filtration rate in outpatients]. AB - The glomerular filtration rate (GFR) calculated by the MDRD formula was estimated in a population of outpatients aged over 18 years. Serum creatinine concentrations were measured, by using 5 analytical systems: Abbott Architect (n = 9054), Roche Modular (n = 22947), Roche Integra 400 (n = 2748), Roche Integra 700 (n = 8350), and Roche Hitachi (n = 20196). For 4 systems, the distribution of GFR differed little. The exception was the Roche Hitachi analytical system where TE and bias were higher than the acceptable requirements. Therefore, with this system, the proportion of outpatients with a GFR of < 60 ml/min per 1.73 m2 was twice higher than that with the use other analytical systems. Such publications should contain data on the analytical quality of measurement of serum creatinine concentrations in the range of 88 to 140 micromol/l. The estimated GFR may be clinically used provided that the performance of this test will be better than the acceptable analytical quality requirements established by the working NKDEP group. PMID- 17918332 TI - [Some lipid and protein metabolic parameters in the Far North migrants]. PMID- 17918333 TI - [Immunological laboratory diagnosis of multiple endocrinopathies (a lecture)]. PMID- 17918334 TI - [Serum albumin binding capacity values in the clinical evaluation of endogenous intoxication in acute viral hepatitis]. AB - The fluorescence technique was used to determine the effective albumin concentration (EAC) and albumin binding reserve (ABR) in 130 patients with acute viral hepatitis A and B in order to establish their values in the evaluation of endogenous intoxication (EI). The most considerable EAC and ABR reduction was found in patients with previous comorbidity and with negative responsiveness changes underlying a high baseline EI level. Despite the resolution of the clinical symptoms of intoxication, the long existence of EAC and ABR changes in the course of the disease in these patients is associated with the tension of the body's key systems in impaired somatic regulation and determines the increased consumption of defense reserves and their rapid exhaustion. Thus, the informative value of the study biochemical parameters of EI increased within the multifactor system of its clinical evaluation. PMID- 17918335 TI - [Neutrophilic granulocytes in of the ovarian cancer progression]. AB - The studies determining phagocytic activity, the levels of myeloperoxidase (MPO) and cationic proteins (CP), and the results of the spontaneous and induced NST tests of peripheral neutrophilic granulocytes (NGC) revealed their functional changes in the course of tumor progression in patients with ovarian cancer. There was an increase in the total count of NGC with simultaneous reductions in the levels of CP, the activity of MPO and in the absorptive capacity of NGC at the late clinical stages of the disease. The results of the NST test did not change. The findings suggest that a growing tumor affects NGC reproduction. PMID- 17918336 TI - [Human embryonic stem cells. Problems and perspectives]. AB - Establishment of human embryonic stem cell lines is one the major achievements in the biological science in the XX century and has excited a wide scientific and social response as embryonic stem cells can be regarded in future as unlimited source of transplantation materials for the replacement cell therapy. To date human embryonic cell lines are obtained in more than 20 countries. In our country the embryonic stem cell researches are carried out in the Institute of Cytology RAS and the Institute of Gene Biology RAS. ESC lines are derived from placed in culture inner cell mass of human preimplantation blastocysts used in the in vitro fertilization procedure. Studies with human ESC go in several directions. Much attention is paid to the elaboration of the optimal conditions for ESC cultivation, mainly to the development of cultivation methods excluding animal feeder cells and other components of animal origin. Another direction is a scale analysis of gene expression specific for the embryonic state of the cells and corresponding signaling pathways. Many efforts are concentrated to find conditions for the directed differentiation of ESC into different tissue-specific cells. It has been shown that ESC are able to differentiate in vitro practically into any somatic cells. Some works are initiated to develop methods for the "therapeutic cloning", that is transfer and reactivation of somatic nuclei into enucleated oocytes or embryonic stem cell cytoblasts. Of great importance is human ESC line standardization. However, the standard requirements for the cells projected for research or therapeutic purposes may be different. It has been found that many permanent human ESC lines undergo genetic and epigenetic changes and, therefore, the cell line genetic stability should be periodically verified. The main aim of the review presented is a detailed consideration of the works analyzing the genetic stability of human and mouse ESC lines. Human ESC lines established in our and as well as in other countries couldn't be used so far in clinical practice. It is highly probable that undifferentiated ESC cannot be applied for therapeutic purposes because of the risk of their malignant transformation. Therefore, main efforts should be focused on the production of progenitor and highly differentiated cells suitable for transplantation derived from ESC. PMID- 17918337 TI - [Chromogranin A: immunocytochemical localization in secretory granules of frog atrial cardiomyocytes]. AB - Chromogranin A (CgA) is a member of the granin family of acidic proteins that present in the secretory granules (SGs) of many endocrine, neuroendocrine and neuronal cells. Atrial natriuretic peptide (ANP)-storing SGs in atrial cardiomyocytes of rat heart also contain CgA. Cardiosuppressive effect of CgA derived peptides (vasostatins) on in vitro isolated and perfused working frog and rat hearts has been shown under both basal conditions and beta-adrenergic stimulation. More recently it has been revealed that rat heart produces and processes CgA-derived vasostatin-containing peptides. Until now nothing has been known about the presence of CgA in an amphibian heart. We have investigated the subcellular localization of CgA in atrial myocytes of adult frog Rana temporaria heart using ultraimmunocytochemical method. Immunocytochemical staining of the frog atrial tissue for CgA and ANP has shown that out of three morphologically different types (A, B and D) of specific cytoplasmic granules (SCGs) present in myocytes only two (A and B)--large (120-200 nm in diameter) granules with more and with less electron dense core--exhibit immunoreactivity (IR) to these two antigens. The third type (D) of granules (80-100 nm in diameter) are small membrane bound granules characterized by highly electron dense core surrounded with a thin halo. These granules revealed negative reaction on immunostaining for both CgA and ANP. The presence of CgA- and ANP-IR in the same SCGs in frog atrial myocytes is consistent with the endocrine nature of these granules. Taking into account our and literature data we propose that CgA present in frog atrial cardiomyocite SCGs might be a precursor of vasostatin-containing peptides, as it takes place in rat heart. It is possible that these CgA-derived peptides together with ANP exert their regulatory function through the autocrine and/or paracrine mechanisms and play important cardioprotective role in frog heart under stress condition. PMID- 17918338 TI - [Differentiation of multipotent mesenchymal stromal cells of bone marrow into cells of cartilage tissue by culturing in three-demential OPLA scaffolds]. AB - Bone marrow multipotent mesenchymal stromal cells represent a perspective material for engineering of human three-dimensional transplants of cartilage tissue. We are demonstrated the opportunity of the directed differentiation of BM MMSC in cells of cartilage tissue by culturing them in three-dimensional scaffolds, presented by polymer OPLA in medium with inductors of chondrogenesis. For loading cells in porous scaffolds used method which essence consist in saturation of polymeric blocks by cellular suspension with the subsequent centrifugal force of cells in scaffolds and culturing of engineering constructs for 28 days in chondrogenic medium. Histological analysis derived in vitro of three-dimensional transplants showed uniform distribution of cells in the matrix with morphologically distinct chondrocytes-like cells of hyaline cartilage. Immunohistochemical analysis detected aggrecan and collagen type II within the extracellular matrix. Preclinical the researches lead on a livestock of immunodeficient mice have shown not toxicity of the engineering constructs. PMID- 17918339 TI - [Reprogramming of nuclear proteasomes in K562 cells undergoing apoptosis. II. Effect of anticancer drug doxorubicin]. AB - The induction of apoptosis in K562 cells by doxorubicin (DR) was used as a model to investigate changes in the subunit composition, phosphorylation state and enzymatic activities of 26S proteasomes in cells undergoing the programmed death. Here we have shown for the first time that proteasomes isolated from the nuclei of control and induced K562 cells differ in their subunit patterns, as well as in the phosphorylation state of subunits on threonine and tyrosine residues. It has been shown for the first time that trypsin- and chymotrypsin-like, and the endoribonuclease activities of nuclear 26S proteasomes are affected under influence of DR on K562 cells. Treatment of K562 cells with DR leads to modification of zeta/alpha5 and iota/alpha6 proteasomal subunits associated with RNase activity of proteasomes. These findings confirm our hypothesis about so called reprogramming of nuclear proteasomes population in undergoing apoptosis K562 cells which is manifested by changes in proteasomal composition, phosphorylation state, and enzymatic activities during the programmed cell death. PMID- 17918340 TI - [Protein kinases abundantly expressed in undifferentiated human ESC lines and derived embryoid bodies]. AB - The ability of human embryonic stem cells (ESCs) to unlimited proliferation and huge differentiation potential makes them very attractive tool both for basic research and biological medicine. There are still little known about mechanisms that govern their differentiation or keep them in a pluripotency state. A variety of signaling events determines gene expression profiles responsible for such mechanisms activation. Protein kinases are key components of the signaling cascades. The knowledge about protein kinases expression profile in undifferentiated ESCs and embryoid bodies (EBs) will allow to understand early differentiation events. We constructed cDNA libraries containing fragments of protein kinases catalytic domain that were expressed in undifferentiated cells or EB of hESM01, hESM02 cell lines. We detected high level of MAK-V expression using Northern-blot hybridization. Semi-quantitative RT-PCR was used to compare the level of abundantly expressed kinases MAK-V, A-RAF-1, MARK3, IGF1R, NEK3 and NEK7 in undifferentiated ESCs or derived EBs. PMID- 17918341 TI - [Mesenchymal stem cells-based therapy of brain ischemic stroke in rat]. AB - Mesenchymal stem cells (MSCs)-based therapy is a promising modern attempt to improve the recovery after stroke. Experiments were carried out on inbred Wistar Kyoto rats. MSCs were isolated, expanded in cultute and labeled with vital fluorescent dye PKH-26. Animals were subjected to middle cerebral artery occlusion (MCAO), followed by injection of 5 x 10(6) rat MSCs into the tail vein 3 days after MCAO. Control group animals received PBS injection (negative control). Therapy results were estimated by the following parameters: behavioral and neurological testing, the brain injure area, the state of damaged region "border" zone and the vessels quantity in the "borden" area. It was shown that control group animals (PBS injection) did not restore their initial behavioral and neurological state, while the experimental group animals (MSCs injection) showed the same parameters as intact rats at 2-3 weeks after MCAO. The size of the damaged region in the control group was approximately 1.5 as large as in the experimental group. The damage in the experimental group was limited to neocortex; caudate nucleus, capsula externa and piriform cortex remained uninjured. Small vessels quantity in the "border" regions was twine higher compared to control group and was approximately equal to an intact brain vessel number. Moreover, it was shown for the first time that after MSCs transplantation the vessels quantity in the neocortex and caudate putamen of contralateral hemisphere was twice as much as in control. We demonstrated that the MSCs transplantation definitely exerted a positive influence upon the brain tissue reparation after stroke. PMID- 17918342 TI - [The influence of sodium nitrite on the osmoresistance of Chinese hamster cells]. AB - We have previously demonstrated that incubation of V-79 cells in the medium containing the nitric oxide donor, NaNO2, increases cell resistance to damaging effect of gamma-rays, UV radiation and hyperthermia. In the present study, we investigated the effects of the nitric oxide donor on the sensitivity of V-79 cells to changes in osmomolarity of the medium by adding different amounts of sodium chloride or water. We found that pretreatment of the cells with NaNO2 resulted in a significant increase in the number of growing cells in 48 h after the treatment. The osmomolarity-dependent morphological changes in cultured cells were also substantially diminished following NaNO2 treatment. This effect could be observed under both hyper- and hypoosmosis, and was dependent on concentration of sodium chloride in hypertonic medium (being maximal under 0.17 M NaCl) and on the amount of water in hypotonic medium (being maximal under 1.1 times the dilution with water). In the experiments with increased osmomolarity, we found that the observed increase in the number of growing cells following NaNO2 treatment was accompanied with a significant increase of the mitotic index. These findings indicate that nitric oxide increases cell resistance to the damaging effects of osmotic shock in the way which is similar to the protective effect of these molecules against radiation and hyperthermia. Similarities in the effects of NaNO2 under different conditions leading to cell damage suggest that nitric oxide might serve as the universal factor participating in recovery of damaged cells and mediating increased cellular resistance to the damaging conditions. PMID- 17918343 TI - [Distribution of cholineacetyltransferase histochemistry in isthmus and medulla of Onchorynchus masu. Tract-tracing observation on the ascending meso-pontine cholinergic system]. AB - The distribution of cholinergic neurons was studies in the brain steam, medulla and rostral spinal cord of the salmon Onchorynchus masu using histochemical choline acetyltransferase (ChAT) detection. Cholinergic neurons were observed in the isthmus, cranial nerve motor nuclei and spinal cord. In order to characterize several cholinergic nuclei observed in the isthmus of O. masu, their projections were studied by application of 1,1'-dioctadecyl-3,3,3',3,' tetramethylindocarbocyanine perchlorate (DiI) to selected structures of the brain. The secondary gustatory nucleus projected mainly to the lateral hypothalamic lobes, whereas the nucleus isthmi projected to the optic tectum and parvocellular superficial pretectal nucleus, as it was earlier described for the other teleost group. In addition, the other isthmic cholinergic nuclei in O. masu may be homologous to the meso-pontine system of mammals. We conclude that the cholinergic systems of teleosts show many primitive features that have been presented during evolution, together with exclusive to the group characteristics. PMID- 17918344 TI - [Simulation of alpha-helix and beta-hairpin formation in water-soluble proteins by the code physics method]. AB - One of the possible ways for complete and final solution of the problem of determination of three-dimensional structure of proteins on amino acid sequence is simulation of protein three-dimensional structure formation. The use of the code physics method developed by the author has been suggested to fulfill this task. The simulation of alpha-helix and beta-hairpin formation in water-soluble proteins as a start of realization of the plan is described here. The results of the simulation were compared with the experimental data for 14 proteins of no more than 50 amino acids and therefore with little number of alpha-helices and beta-strands (to meet limits of simulation process) and with secondary structure predictions by the best to data methods of protein secondary structure prediction, PSIpred, PORTER and PROFsec. Secondary structure of the proteins, obtained as a result of the simulation of alpha-helix and beta-hairpin formation using the code physics method, corresponded completely to experimental data while the secondary structure predicted by the PSIpred, PORTER and PROFsec methods differed from these data significantly. PMID- 17918345 TI - [Karyological investigations of populations of the lizard Zootoca vivipara (Juaquin, 1787) from the Baltic Sea basin (western region of Russia)]. AB - Populations of the lizard Zootoca vivipara (Juaquin, 1787) (Lacertidae, Sauria) from Western and Central Europe are poorly differentiated in morphology. However, they differ from each other by several karyotype markers and their reproductive mode (oviparous and viviparous types). Here we report for the first time the results of combined investigations of Z. vivipara from Eastern Europe (the Baltic Sea basin of Western Russia). The karyotype and reproductive mode of the specimens from four previously not examined populations in Kaliningrad region have been studied. It has been shown that these lizards have viviparous mode of reproduction and their karyotypes vary on morphology of W-sex chromosomes. Females possess 2n = 35 : 32 A + Z1Z2W, with W--SV and 2n = 35 : 32 A + Z1Z2W, with W--A/ST. We have identified these populations karyologically and have first revealed that they belong to two different viviparous forms of nominative subspecies Z. v. vivipara, namely to its "western" and "Russian (eastern)" forms. These data have confirmed that the Baltic Sea basin in Eastern Europe is a zone of secondary contact of these chromosomal forms of Z. v. vivipara. The results obtained do not now allow us to speak about their sympatry, parapatry and (or) hybridization events. The results suggest that during postglasial time "western" form might colonize this region from western Europe whereas "Russian (eastern)" form might do it from Southern-Eastern Europe. It has been recommended to save the biodiversity of Z. vivipara in Kaliningrad region, namely the peripheral and very rare on vast territory of Russia "western" form and unique for western region of Russia "Russian (eastern)" form. PMID- 17918346 TI - [Protein glycosylation in the spores of the microsporidia Paranosema (Antonospora) grylli]. AB - Long adaptation of microsporidia, a large group of fungi-related protozoa, to intracellular lifestyle has resulted in drastic minimization of a parasite cell. Thus, diversity of carbohydrates in microsporidia glycoproteins and proteoglycans is expected to be restricted by O-linked manno-oligosaccharides because three genes involved in O-mannosylation of proteins and no components of N-linked glycosylation machinery were found in genome of human pathogen Encephalitozoon cuniculi. In this study we investigated glycosylation of spore proteins of microsporidia Paranosema (Antonospora) grylli infecting crickets Gryllus bimaculatus. Using periodic acid-Shiff reagent staining we have demonstrated that some P. grylli spore proteins are highly-glycosylated. The major polar tube protein (PTP1) of 56 kDa was shown as the most intensively decorated band. The experiments with N-glycosidase F and WGA lectin did not reveal any N-glycosylated proteins in P. grylli spores. At the same time, incubation of major spore wall protein of 40 kDa (p40) with mannose specific lectin GNA resulted in specific binding that was reduced by pretreatment of the protein with mannosidases. Interestingly, in spite of PTP1 glycosylation, polar tube proteins extracted from P. grylli spores were not precipitated by GNA-agarose. Since P. grylli and E. cuniculi are distantly related, our data suggest that dramatic reduction of protein glycosylation machinery is a common feature of microsporidia. PMID- 17918347 TI - [DNA damages in patients having occupational allergic dermatoses]. AB - Studies of DNA damage processes identified high percentage of antibodies to single- and double-stranded DNA in patients having occupational allergic dermatoses,demonstrated successive increase of antibodies to single- and double stranded DNA and to nucleosoma with longer length of service. PMID- 17918348 TI - [Contemporary hygienic requirements to work conditions of medical professionals performing ultrasound investigations]. AB - The article covers work conditions and health state of medical professionals performing ultrasound investigations on modern medical diagnostic equipment. The authors suggest a guidelines project including complex of measures to prevent unfavorable influence of contact ultrasound on medical professionals. PMID- 17918350 TI - Effects of Acanthocephalus lucii (Acanthocephala) on intermediate host survival and growth: implications for exploitation strategies. AB - Intermediate host exploitation by parasites is presumably constrained by the need to maintain host viability until transmission occurs. The relationship between parasitism and host survival, though, likely varies as the energetic requirements of parasites change during ontogeny. An experimental infection of an acanthocephalan (Acanthocephalus lucii) in its isopod intermediate host (Asellus aquaticus) was conducted to investigate host survival and growth throughout the course of parasite development. Individual isopods were infected by exposure to fish feces containing parasite eggs. Isopods exposed to A. lucii had reduced survival, but only early in the infection. Mean infection intensity was high relative to natural levels, but host mortality was not intensity dependent. Similarly, a group of naturally infected isopods harboring multiple cystacanths did not have lower survival than singly infected isopods. Isopods that were not exposed to the parasite exhibited sexual differences in survival and molting, but these patterns were reversed or absent in exposed isopods, possibly as a consequence of castration. Further, exposed isopods seemed to have accelerated molting relative to unexposed controls. Infection had no apparent effect on isopod growth. The effects of A. lucii on isopod survival and growth undermine common assumptions concerning parasite-induced host mortality and the resource constraints experienced by developing parasites. PMID- 17918349 TI - Comparison of the peptidase activity in the oncosphere excretory/secretory products of Taenia solium and Taenia saginata. AB - We compared the peptidase activities of the excretory/secretory (E/S) antigens of oncospheres of Taenia solium and related, but nonpathogenic, Taenia saginata. Taenia solium and T. saginata oncospheres were cultured, and the spent media of 24-, 48-, 72-, and 96-hr fractions were analyzed. Activities for serine peptidases (chymotrypsin-, trypsin-, and elastase-like), cysteine peptidases (cathepsin B-, cathepsin L-, and calpaine-like), and aminopeptidase (B-like peptidases) were tested fluorometrically with peptides coupled to 7-amino-4 methylcoumarin. In both species, the E/S antigens showed cysteine, serine, and aminopeptidase activities. Although no particular peptidase had high activity in T. solium, and was absent in T. saginata, or vice versa, different patterns of activity were found. A chymotrypsin-like peptidase showed the highest activity in both parasites, and it had 10 times higher activity in T. solium than in T. saginata. Trypsin-like and cathepsin B-like activities were significantly higher in T. solium. Minimal levels of cathepsin B were present in both species, and higher levels of elastase-like and cathepsin L-like activity were observed in T. saginata. Taenia solium and T. saginata have different levels and temporal activities of proteolytic enzymes that could play a modulator role in the host specificity for larval invasion through penetration of the intestinal mucosa. PMID- 17918352 TI - Field evidence of host size-dependent parasitism in two manipulative parasites. AB - The distribution of parasites within host natural populations has often been found to be host age-dependent. Host mortality induced by parasites is the commonest hypothesis proposed for explaining this pattern. Despite its potential importance in ecology, the parasitism intensity in relation with the host age has rarely been studied in the field. The 2 manipulative acanthocephalans, Polymorphus minutus and Pomphorhynchus laevis, use the amphipod Gammarus pulex as an intermediate host, and their infection intensity and incidence among G. pulex populations were examined by analyzing 2 large samples of hosts collected in eastern France. Both parasites had low prevalence in the host populations, but their mean abundances were highly related with gammarid age. For the 2 acanthocephalans, results reported a disappearance or an absence of heavily infected hosts in the older host age classes. These results suggested that parasites that alter intermediate host behavior for enhancing their transmission success to the definitive host reduce the survival of their intermediate host. In conclusion, manipulative parasites might act as a mechanism regulating the density of gammarid populations. PMID- 17918351 TI - Proximate factors affecting the larval life history of Acanthocephalus lucii (Acanthocephala). AB - The growth and eventual size of larval helminths in their intermediate hosts presumably has a variety of fitness consequences. Therefore, elucidating the proximate factors affecting parasite development within intermediate hosts should provide insight into the evolution of parasite life histories. An experimental infection that resulted in heavy intensities of an acanthocephalan (Acanthocephalus lucii) in its isopod intermediate host (Asellus aquaticus) permitted the examination of parasite developmental responses to variable levels of resource availability and intraspecific competition. Isopods were infected by exposure to egg-containing fish feces, and larval infrapopulations were monitored throughout the course of A. lucii development. The relative rate of parasite growth slowed over time, and indications of resource constraints on developing parasites, e.g., crowding effects, were only observed in late infections. Consequently, the factors likely representative of resource availability to larval parasites (host size and molting rate) primarily affected parasite size in late infections. Moreover, at this stage of infection, competitive interactions, gauged by variation in worm size, seemed to be alleviated by greater resources, i.e., larger hosts that molted more frequently. The relatively rapid, unconstrained growth of young parasites may be worse for host viability than the slower, resource-limited growth of larger parasites. PMID- 17918353 TI - Helminth communities in five species of sympatric amphibians from three adjacent ephemeral ponds in southeastern Wisconsin. AB - Representatives of 5 amphibian species (313 individuals), including eastern American toads (Bufo americanus), wood frogs (Rana sylvatica), spring peepers (Pseudacris crucifer), blue-spotted salamanders (Ambystoma laterale), and central newts (Notophthalmus viridescens louisianensis), were collected from 3 ephemeral ponds during spring 1994, and they were inspected for helminth parasites. The component communities of anurans were more diverse than those of caudates. Infracommunities of all host species were isolationist and depauperate, due mostly to host ectothermy and low vagility. Toad infracommunities were dominated by skin-penetrating nematodes, and they had the highest values of mean total parasite abundance, mean species richness, and overall prevalence. This was likely due to their greater vagility compared with other host species. Infracommunities of wood frogs and blue-spotted salamanders had intermediate values for these measures of parasitism, whereas spring peeper and newt infracommunities had the lowest values. In addition to relative vagility, feeding habits and habitat preference were likely important in helminth community structure. Body size also seemed to play a role because mean wet weight of host species followed the same general trend as values of parasitism. However, effects of size were variable within host species and difficult to separate from other aspects of host ecology. PMID- 17918354 TI - New and previously described species of Dactylogyridae (Monogenoidea) from the gills of Panamanian freshwater fishes (Teleostei). AB - During an investigation of the diversity of metazoan parasites of 7 freshwater fish species from 3 localities in central Panama, the following gill dactylogyrid (Monogenoidea) species were found: Aphanoblastella chagresii n. sp. from Pimelodella chagresi (Heptapteridae); Aphanoblastella travassosi (Price, 1938) Kritsky, Mendoza-Franco, and Scholz, 2000 from Rhamdia quelen (Heptapteridae); Diaphorocleidus petrosusi n. sp. from Brycon petrosus (Characidae); Gussevia asota Kritsky, Thatcher, and Boeger, 1989, from Astronotus ocellatus (Cichlidae); Sciadicleithrum panamensis n. sp. from Aequidens coeruleopunctatus (Cichlidae); Urocleidoides flegomai n. sp. from Piabucina panamensis (Lebiasinidae); and Urocleidoides similuncus n. sp. from Poecilia gillii (Poeciliidae). Consideration of the comparative morphology and distribution of these parasites along with the evolutionary history of the host fishes suggests that diversification may be associated with geotectonic events that provided isolation of the Central American fauna with the uplift of the Panamanian Isthmus during early Pliocene (3 mya). PMID- 17918355 TI - Three new species of Ligophorus (Monogenea: Dactylogyridae) on the gills of Mugil cephalus (Teleostei: Mugilidae) from the Japan Sea. AB - Comparative morphology and multivariate morphometric analysis of monogeneans collected on flathead mullets Mugil cephalus from 2 Russian localities of the Japan Sea revealed the presence of 3 new species of Ligophorus, namely, L. domnichi n. sp., L. pacificus n. sp., and L. cheleus n. sp., which are described herein. So far, only 1 species of dactylogyrid monogenean identified as Ligophorus chabaudi was known on flathead mullets in this sea, but after comparison with the present material, we propose that this form actually represents L. domnichi n. sp. Results support previous zoogeographical evidence, suggesting that flathead mullets from different seas harbor different species complexes of Ligophorus. One interesting finding is that the 3 new species have a U-shaped ovary, whereas ovate ovaries have been reported in previous descriptions of species of the genus, e.g., L. vanbenedenii, L. parvicirrus, L. imitans, and L. chongmingensis. The U-shaped ovary was revealed only when the worms were observed in lateral view. The additional examination of L. vanbenedenii, L. parvicirrus, L. imitans, and L. pilengas specimens from our collections also revealed a U-shaped ovary in these forms as well. Further studies should establish whether or not this character is shared by all members of the genus. PMID- 17918356 TI - Two new species of Haliotrema (Monogenoidea: Dactylogyridae) from Acanthurus nigrofuscus and Acanthurus olivaceus (Teleostei: Acanthuridae) in the South China Sea. AB - Haliotrema leporinus n. sp. and Haliotrema pratasensis n. sp. are described from the gills of surgeonfishes (Acanthuridae) off the Pratas Islands in the South China Sea. Haliotrema leporinus n. sp. differs from all other members of the genus by possessing a rabbit-head-shaped sclerotized piece in the haptor. Haliotrema pratasensis n. sp. differs from its congeners by having a copulatory organ with a clockwise coil and a funnel-shaped base and by lacking an accessory piece. PMID- 17918357 TI - The first chondracanthid (Copepoda: Cyclopoida) reported from cultured finfish, with a revised key to the species of Chondracanthus. AB - A new species of the Chondracanthidae (Copepoda: Cyclopoida), Chondracanthus goldsmidi, is described based on material collected from the nasobranchial region of striped trumpeter (Latris lineata [Forster]) cultured at the Tasmanian Aqua culture and Fisheries Institute, Marine Research Laboratories, Australia. This represents the first report of a chondracanthid copepod infecting cultured finfish and the first metazoan parasite from cultured striped trumpeter. Chondracanthus goldsmidi n. sp. can be distinguished from its female congeners by the absence of lateral processes on the head and the presence of 3 pairs of lateral trunk outgrowths, 3 middorsal body outgrowths (of which the first 2 are rounded), a small and subcylindrical antennule, and unornamented legs 1 and 2. A revised key to the 39 valid species of Chondracanthus is provided. PMID- 17918358 TI - Impact of population structure on genetic diversity of a potential vaccine target in the canine hookworm (Ancylostoma caninum). AB - Ancylostoma caninum is a globally distributed canine parasitic nematode. To test whether positive selection, population structure, or both affect genetic variation at the candidate vaccine target Ancylostoma secreted protein 1 (asp-1), we have quantified the genetic variation in A. caninum at asp-1 and a mitochondrial gene, cytochrome oxidase subunit 1 (cox-1), using the statistical population analysis tools found in the SNAP Workbench. The mitochondrial gene cox 1 exhibits moderate diversity within 2 North American samples, comparable to the level of variation observed in other parasitic nematodes. The protein coding portion for the C-terminal half of asp-1 shows similar levels of genetic variation in a Wake County, North Carolina, sample as cox-1. Standard tests of neutrality provide little formal evidence for selection acting on this locus, but haplotype networks for 2 of the exon regions have significantly different topologies, consistent with different evolutionary forces shaping variation at either end of a 1.3-kilobase stretch of sequence. Evidence for gene flow among geographically distinct samples suggests that the mobility of hosts of A. caninum is an important contributing factor to the population structure of the parasite. PMID- 17918359 TI - Evaluation of two Toxoplasma gondii serologic tests used in a serosurvey of domestic cats in California. AB - We evaluated the sensitivity (Se) and specificity (Sp) of an IgG enzyme-linked immunosorbent assay (ELISA) and IgG indirect fluorescent antibody test (IFAT) for detection of Toxoplasma gondii-specific antibodies in sera from 2 cat populations using a Bayesian approach. Accounting for test covariance, the Se and Sp of the IgG ELISA were estimated to be 92.6% and 96.5%, and those of the IgG IFAT were 81.0% and 93.8%, respectively. Both tests performed poorly in cats experimentally coinfected with feline immunodeficiency virus and T. gondii. Excluding this group, Se and Sp of the ELISA were virtually unchanged (92.3% and 96.4%, respectively), whereas the IFAT Se improved to 94.2% and Sp remained stable at 93.7%. These tests and an IgM ELISA were applied to 123 cat sera from the Morro Bay area, California, where high morbidity and mortality attributable to toxoplasmosis have been detected in southern sea otters. Age-adjusted IgG seroprevalence in this population was estimated to be 29.6%, and it did not differ between owned and unowned cats. Accounting for Se, Sp, and test covariances, age-adjusted seroprevalence was 45.0%. The odds for T. gondii seropositivity were 12.3-fold higher for cats aged >12 mo compared with cats aged <6 mo. PMID- 17918360 TI - Identification of Fasciola hepatica recombinant 15-kDa fatty acid-binding protein T-cell epitopes that protect against experimental fascioliasis in rabbits and mice. AB - Vaccination with fatty acid-binding proteins (FABPs) from Fasciola hepatica has been shown to confer significant levels of protection against challenge infection in mice, rabbits, and sheep. A recombinant 15-kDa FABP (rFh15) has been purified and also shown to be an immunoprotective molecule. From the rFh15 molecule sequence 2, 12- and 10-mer putative T-cell epitopes were identified, the first an Fh15Ta of amino acid sequence IKMVSSLKTKIT, and the second an Fh15Tb of amino acid sequence VKAVTTLLKA. The synthesized oligonucleotides were cloned individually into a pGEX-2TK expression vector. The overexpressed fusion protein was affinity purified using glutathione S-transferase (GST) by competitive elution with excess reduced glutathione. These GST fusion proteins were emulsified in Freund adjuvant for rabbit immunizations or further purified as peptides after digestion with thrombin. The purified 12- and 10-mer peptides were either emulsified in Freund adjuvant for immunizations in rabbits or used in an adjuvant-adaptation (ADAD) system, followed by challenge infection with F. hepatica metacercariae in mice and rabbits. In vaccinated-challenged rabbits, the highest levels of protection were found in those treated with GST-epitopes (Fh15Ta 48.2% and Fh15Tb 59.1% reduction, respectively), as compared to GST immunized controls. Moreover, those immunized with Fh15Ta had higher (84%) numbers of immature flukes as compared with Fh15Tb (41%) or GST alone (64%). The rabbits immunized with the putative T-cell epitopes in adjuvant had a 13% reduction in flukes in those with Fh15Ta and also were highest with immature flukes (46%). In vaccinated mice challenged with a lethal number of metacercariae, both CD-1 and BALB/c mice treated with complete ADAD-GST-Ta had the highest (40%) survival rates of all groups by 47 days postinfection. Thus the Fh15Ta and Fh15Tb polypeptide epitopes warrant further study as a potential vaccine against F. hepatica. Antibody isotype studies in mice revealed a mixed Thl/Th2 response to vaccination. PMID- 17918361 TI - Renewed hope for a vaccine against the intestinal adult Taenia solium. AB - Review of experimental and observational evidence about various cestode infections of mammalian hosts revives hope for the development of an effective vaccine against adult intestinal tapeworms, the central protagonists in their transmission dynamics. As for Taenia solium, there are abundant immunological data regarding cysticercosis in humans and pigs, but information about human taeniasis is scarce. A single publication reporting protection against T. solium taeniasis by experimental primo infection and by vaccination of an experimental foster host, the immunocompetent female hamster, kindles the hope of a vaccine against the tapeworm to be used in humans, its only natural definitive host. PMID- 17918362 TI - Surface membrane proteins of Biomphalaria glabrata embryonic cells bind fucosyl determinants on the tegumental surface of Schistosoma mansoni primary sporocysts. AB - Previous observations that in vitro adherence of Biomphalaria glabrata embryonic (Bge) cells to sporocyst larval stages of Schistosoma mansoni was strongly inhibited by fucoidan, a sulfated polymer of L-fucose, suggested a role for lectinlike Bge cell receptors in sporocyst binding interactions. In the present investigation, monoclonal antibodies with specificities to 3 major glycan determinants found on schistosomes, LacdiNAc, fucosylated LacdiNAc (LDNF), and the Lewis X antigen, were used in adhesion blocking studies to further analyze the molecular interactions at the host-parasite interface. Results showed that only the anti-LDNF antibody significantly reduced snail Bge cell adhesion to the surface of sporocysts, suggesting that fucosyl determinants may be important in larval-host cell interactions. Affinity chromatographic separation of fucosyl reactive Bge cell proteins from fucoidan-bound Sepharose 4B revealed the presence of polypeptides ranging from 6 to 200 kDa after elution with fucoidan-containing buffer. Pre-elution of the Bge protein-bound affinity column with dextran (Dex) and dextran sulfate (DexS) before introduction of the fucoidan buffer served as controls for protein binding based on nonspecific sugar or negative charge interactions. A subset of polypeptides (approximately 35-150 kDa) released by fucoidan elution was identified as Bge surface membrane proteins, representing putative fucosyl-binding proteins. Far-western blot analysis also demonstrated binding reactivity between Bge cell and sporocyst tegumental proteins. The finding that several of these parasite-binding Bge cell proteins were also fucoidan-reactive suggests the possible involvement of these molecules in mediating cellular interactions with sporocyst tegumental carbohydrates. It is concluded that Bge cells have surface protein(s) that may be playing a role in facilitating host cell adhesion to the surface of schistosome primary sporocysts through larval fucosylated glycoconjugates. PMID- 17918363 TI - Redescription of two Dicyemennea (Phylum: Dicyemida) from Rossia pacifica (Mollusca: Cephalopoda: Decapoda). AB - Two species of dicyemid mesozoan are redescribed from Rossia pacifica Berry, 1911, collected off Iwase in Toyama Bay, Honshu, Japan. Dicyemennea brevicephaloides Bogolepova-Dobrokhotova, 1962, is a large species that reaches about 4,000 microm in length. This species attaches to the surface of the branchial heart appendages. The vermiform stages are characterized as having 23 peripheral cells, a disc-shaped calotte, and an axial cell that extends to the propolar cells. An anterior abortive axial cell is absent in vermiform embryos. Infusoriform embryos consist of 35 cells; a single nucleus is present in each urn cell and the refringent bodies are absent. Dicyemennea rossiae Bogolepova Dobrokhotova, 1962, is a medium species that reaches about 2000 microm in length. This species lives in folds of the renal appendages. The vermiform stages are characterized as having 29-34 peripheral cells, a conical calotte, and an axial cell that extends to the middle of the metapolar cells. An anterior abortive axial cell is present in vermiform embryos. Infusoriform embryos consist of 39 cells; 2 nuclei are present in each urn cell and the refringent bodies are solid. PMID- 17918364 TI - Chimpanzee pinworm, Enterobius anthropopitheci (Nematoda: Oxyuridae), maintained for more than twenty years in captive chimpanzees in Japan. AB - The chimpanzee pinworm, Enterobius anthropopitheci (Gedoelst, 1916), was found in chimpanzees, Pan troglodytes, reared in Kumamoto Primate Research Park, Sanwa Kagaku Kenkyusho Co., Ltd., Kumamoto, Japan, in 2006. Because the chimpanzees in this institution originated from chimpanzees imported from Africa before 1984, it is considered that E. anthropopitheci infection has persisted for more than 20 yr in the chimpanzees. Analysis of pinworm specimens preserved in the institution revealed that transition of predominant pinworm species occurred, responding to the change of anthelmintics used for pinworm treatment. Present dominance of E. anthropopitheci is surmised to be caused by fenbendazole, which has been adopted from 2002. Scarcity of mixed infection with E. anthropopitheci and Enterobius vermicularis suggests interspecific competition between the pinworms. PMID- 17918366 TI - A new genus and species of Macroderoididae, and other digeneans from fishes of Lake Malawi, Africa. AB - Malawitrema staufferi n. gen., n. sp., an unusual digenean, is described from Clarias mossambicus (type host) and Bagrus meridionalis from Lake Malawi. It has a small, pyriform body, with a spinous tegument. The ceca are relative short, not reaching to the testes. The 2 testes are symmetrical in the middle hind body. The cirrus sac is long and narrow, reaching into hind body. The genital pore is median, immediately anterior to the ventral sucker. The ovary is pretesticular, and a canalicular seminal vesicle and Laurer's canal are present. The uterus usually reaches distinctly posteriorly to testes. The eggs are small. The follicular vitellarium is in 2 small fields just anterior to testes. The ventrally subterminal excretory pore leads to a claviform vesicle. This species does not fit clearly into any known family of digeneans and is placed in the Macroderoididae as a temporary measure. Other digeneans reported from Lake Malawi include Haplorchoides cahirinus (Looss, 1896) in C. mossambicus and B. meridionalis; Astiotrema turneri Bray, van Oosterhout, Blais & Cable, 2006 in Protomelas annectens, P. cf. taeniolatus, Labeotropheus fuelleborni, Ctenopharynx (Otopharynx) pictus, and Pseudotropheus zebra; Glossidium pedatum Looss, 1899 in C. mossambicus and B. meridionalis; and an unidentifiable sanguinicolid from Bathyclarias nyasensis. PMID- 17918365 TI - Development and pathology of Echinostoma caproni in experimentally infected mice. AB - In the present article, several parasitological features of mice, each experimentally infected with 75 metacercariae of Echinostoma caproni (Trematoda: Echinostomatidae), were studied during the first 12 wk postinfection. Moreover, the early pathological responses also were analyzed and compared with data previously published on other host species of E. caproni to gain further insight into the factors determining worm rejection or establishment of chronic infections. The results obtained show that the pattern of E. caproni infection in mice is consistent with a highly compatible host-parasite system. This combination is characterized by a high worm establishment, high egg output, and long survival of the worms. However, some differences with respect to other highly compatible hosts have been observed, particularly in relation to the survival of the adult worms. Histological studies suggest that the kinetics of goblet cells, mucosal neutrophils, and mononuclear inflammatory cells in the mesentery seem to be essential in determining the course of E. caproni infection in mice. PMID- 17918367 TI - New species of Skrjabinodon (Nematoda: Pharyngodonidae) in Uracentron flaviceps (Squamata: Iguanidae) from Ecuador and Peru. AB - Skrjabinodon dixoni n. sp. from the large intestine of Uracentron flaviceps (Squamata: Iguanidae) from Peru is described and illustrated. It is also reported in the same host from Ecuador. Skrjabinodon dixoni n. sp. differs from other species assigned to Skrjabinodon by morphology of tail filament and number of tail filament spines. PMID- 17918368 TI - Redescription, synonymy, and new records of Vexillata noviberiae (Dikmans, 1935) (Nematoda: Trichostrongylina), a parasite of rabbits Sylvilagus spp. (Leporidae) in the United States. AB - Vexillata noviberiae (Dikmans, 1935) (Trichostrongylina: Heligmosomoidea), originally described as a parasite of Sylvilagus floridanus from Louisiana, is redescribed from material collected from Sylvilagus palustris in Florida and from S. floridanus in Kansas. New morphometric and morphological data are provided. Stunkardionema halla Arnold, 1941, described from S. floridanus from Kansas and New York, is proposed as a junior synonym of V. noviberiae. These findings confirm the occurrence of V. noviberiae as a parasite of rabbits and its wide distribution range in North America. PMID- 17918369 TI - A collection of fish leeches (Hirudinida: Piscicolidae) from Japan and surrounding waters, including redescriptions of three species. AB - Leeches were observed incidentally on fishes in collections made from 1975 to 2006 in Japan and surrounding waters, or from mariculture facilities or public aquaria in Japan. Seven species of leeches in 7 genera were collected- Crangonobdella maculosa, Johanssonia arctica, Limnotrachelobdella okae, Platybdella olriki, Stibarobdella bimaculata, Taimenobdella amurensis, and Trachelobdella livanovi. The transfer of Calliobdella livanovi to Trachelobdella is supported, and Trachelobdella livanovi and Taimenobdella amurensis are redescribed based on new specimens. Stibarobdella bimaculata is synonymized with Stibarobdella macrothela based on eyes, tubercle patterns, and sucker size ratios. Taimenobdella amurensis, C. maculosa, J. arctica, S. macrothela, and P. olriki are reported for the first time from Japan. New hosts are reported for L. okae, T. livanovi, S. macrothela, C. maculosa, J. arctica, and P. olriki. Stibarobdella moorei was not collected during this study, but a well-preserved specimen collected in Japan was discovered in the Museum National d'Histoire Naturelle, Paris, France, and it allowed a redescription of this species. Stibarobdella loricata is synonymized with S. moorei based on tubercle patterns and the presence of papillae and a marginal fringe on the oral sucker. PMID- 17918370 TI - Paracosmocercella rosettae n. gen. et n. sp. (Nematoda: Cosmocercoidea: Cosmocercidae) collected from the Japanese tree frog, Hyla japonica (Anura: Hylidae), in Japan. AB - A new genus, Paracosmocercella (Nematoda: Cosmocercoidea: Cosmocercidae), is proposed with monotypic Paracosmocercella rosettae n. sp. from the Japanese tree frog, Hyla japonica Gunther, 1859, in Oita, Kyushu Island, Japan. Paracosmocercella resembles Cosmocercella and Cosmocercoides by having 2 rows of rosette papillae without plectanes ventrally in preanal region of male, but it is readily distinguished from Cosmocercella by lacking clear vesicle supporting rosette papillae and from Cosmocercoides by lacking rosette papillae in perianal region and having large-sized eggs, which hatch in uterus. PMID- 17918371 TI - Diversity and phylogeny of mitochondrial cytochrome B lineages from six morphospecies of avian Haemoproteus (Haemosporida: Haemoproteidae). AB - Species of Haemoproteus (Haemosporida: Haemoproteidae), avian haemosporidians, have traditionally been described based on morphology of their gametocytes and on limited experimental information on their vertebrate host specificity. We investigated to what extent the morphological species are represented by monophyletic groups based on DNA sequence data using 2 different fragment lengths of the cytochrome b (cyt. b) gene. Phylogenetic reconstructions of obtained cyt. b lineages from 6 morphospecies of Haemoproteus showed that all lineages formed monophyletic clusters matching the morphospecies. Comparing our data with a recently published study showed that this is not always the case; the morphospecies H. belopolskyi consists of 2 distinct clusters of lineages that apparently have converged in morphology. However, the overall broad congruence between the molecular and morphological clustering of lineages will facilitate the integration of the knowledge obtained by traditional and molecular parasitology. Mean between morphospecies variation was 10-fold higher than the within species variation (5.5% vs. 0.54%), suggesting that Haemoproteus lineages with a genetic differentiation >5% are expected to be morphologically differentiated in most cases. When investigate the utility of 2 different fragment sizes of the cyt. b gene, the partial, 479-bp, cyt. b protocol picked up all mitochondrial (mt)DNA lineages that are found when using the full cyt. b gene, 1073 bp, suggesting that this protocol is sufficient for identification of most mtDNA lineages. All of the mtDNA lineages were associated with unique alleles when amplification was possible at a nuclear locus, strengthening the hypothesis that the designation of lineages based on mtDNA is largely genome-wide representative. We, therefore, propose the use of a cyt. b fragment of this length as a standard gene fragment for a DNA bar-coding system for avian Haemoproteus species. PMID- 17918372 TI - A new species of Macrobothriidae (Cestoda: Diphyllidea) from thornback ray Platyrhina sinensis in China. AB - A new cestode, Macrobothridium sinensis n. sp., is described from the spiral intestine of Platyrhina sinensis from coast of Xiamen, China. It is the first record of the order Diphyllidea in China. The new species can be distinguished from the 3 previously described species of Macrobothridium by the testes number (16-24 vs. 29-37, 27-46, and 5-6, in M. rhynchobati, M. euterpes, and M. syrtensis, respectively) and a single-row arrangement of testes; M. sinensis is most similar to M. euterpes in strobila length (1.77-6.23 vs. 2-4.5 mm) and number of segments (6-8 vs. 5-9). In addition, M. sinensis differs from M. rhynchobati and M. syrtensis in the strobila length, ovary shape, and number of apical hooks. The uterine pore is the first described in the Diphyllidea; thus, the present uterine pore can now be used to distinguish Macrobothridium from Echinobothrium and Ditrachybothridium in the Diphyllidae. PMID- 17918373 TI - A new species of Comephoronema (Nematoda: Cystidicolidae) from the stomach of the abyssal halosaur Halosauropsis macrochir (Teleostei) from the Mid-Atlantic Ridge. AB - A new species of parasitic nematode Comephoronema macrochiri n. sp. (Cystidicolidae), is described from the stomach of the marine deep-sea fish Halosauropsis macrochir (abyssal halosaur) from the Mid-Atlantic Ridge (MAR). The new species, studied with both light and scanning electron microscopy, is characterized mainly by 6 pairs of preanal papillae, by which it principally differs from members of Ascarophis; the spicules are 297-375 microm and 99-120 microm long and fully developed eggs possess 2 long filaments on 1 pole. Rhabdochona beatriceinsleyae is transferred to Comephoronema as C. beatriceinsleyae (Holloway and Klewer, 1969) n. comb. Comephoronema macrochiri differs from all other congeners mainly in having eggs with filaments on 1 pole only, and from individual species by some additional features such as the number of preanal papillae, the shape of pseudolabial projections, and the body and organ measurements. PMID- 17918374 TI - A new species of Paracapillaria (Nematoda: Capillariidae) from the intestine of the toad Duttaphrynus melanostictus (Anura) from the Malayan Peninsula. AB - A new species of parasitic nematode, Paracapillaria malayensis n. sp. (Capillariidae), is described from the small intestine of the toad Duttaphrynus melanostictus imported from the Malayan Peninsula to the Czech Republic. The new species differs from the only other congeneric species, Paracapillaria spratti, mainly in the shape and structure of the spicular proximal end (with a lobular rim), smaller eggs (45-51 x 21-24 microm), longer spicule (336 microm), and the number (37-38) of stichocytes in gravid females; whereas P. spratti parasitizes frogs of the Microhylidae in Papua New Guinea, P. malayensis is a parasite of Bufonidae in the Malayan Peninsula. Other Paracapillaria spp. are parasites of fishes, birds, or mammals and they mostly differ from P. malayensis in the structure of eggs and some other morphological features. PMID- 17918375 TI - Two new species of cosmocercids (Ascaridida) in the toad Chaunus arenarum (Anura: Bufonidae) from Argentina. AB - Aplectana tarija n. sp. and Cosmocercoides lilloi n. sp., intestinal parasites of the toad Chaunus arenarum, are described. Of the 41 nominal species of Aplectana, A. tarija n. sp. represents the 22nd Neotropical species and the second species reported from Argentina. Of the 14 nominal species of Cosmocercoides, C. lilloi n. sp. represents the 15th species and the first species reported from South America. PMID- 17918376 TI - New species of Haplometroides (Digenea: Plagiorchiidae) from Phalotris nasutus (Gomes, 1915) (Serpentes, Colubridae). AB - A new species of Haplometroides (Digenea, Plagiorchiidae) is described from a specimen of Phalotris nasutus (Gomes, 1915) (Serpentes, Colubridae). The host snake was obtained in the municipality of Corumbd, Mato Grosso do Sul State, Brazil. Trematodes were recovered from esophagus, stomach, and small intestine of the host. The main characteristic of the new species is the vitellaria, which is intercecal, cecal, and extracecal in the preacetabular region. A key for identification of the species in Haplometroides is proposed. PMID- 17918378 TI - Multiplication of Trypanosoma pacifica (Euglenozoa: Kinetoplastea) in English sole, Parophrys vetulus, from Oregon coastal waters. AB - Multiplication of Trypanosoma pacifica was common in the fish host from observations of live flagellates and Giemsa-stained blood smears. Multiplication began with the elongation of the kinetoplast, thickening of the posterior portion of the body, and appearance of a new flagellum near the kinetoplast. The new flagellum was very rigid when less than 3 microm in length, but it became flexible as it elongated. When the new flagellum was approximately 12 microm in length, cell division began and the kinetoplast also began to divide. The timing of nuclear division was variable. Generally, it did not occur until division of the kinetoplast had begun, but occasionally binucleate individuals were observed before cell or kinetoplast division was apparent. As division continued, 1 nucleus migrated past the dividing kinetoplast into the future daughter trypanosome. Finally, the kinetoplast completed division and the trypanosomes separated. Cell division was unequal, with the daughter trypanosome being smaller than the parent and with a more weakly developed undulating membrane. PMID- 17918377 TI - Chemical inactivation of Toxoplasma gondii oocysts in water. AB - The protozoan parasite Toxoplasma gondii is increasingly recognized as a waterborne pathogen. Infection can be acquired by drinking contaminated water and conventional water treatments may not effectively inactivate tough, environmentally resistant oocysts. The present study was performed to assess the efficacy of 2 commonly used chemicals, sodium hypochlorite and ozone, to inactivate T. gondii oocysts in water. Oocysts were exposed to 100 mg/L of chlorine for 30 min, or for 2, 4, 8, 16, and 24 hr, or to 6 mg/L of ozone for 1, 2, 4, 8, or 12 min. Oocyst viability was determined by mouse bioassay. Serology, immunohistochemistry, and in vitro parasite isolation were used to evaluate mice for infection. Initially, mouse bioassay experiments were conducted to compare the analytical sensitivity of these 3 detection methods prior to completing the chemical inactivation experiments. Toxoplasma gondii infection was confirmed by at least 1 of the 3 detection methods in mice inoculated with all doses (10(5) 10(0)) of oocysts. Results of the chemical exposure experiments indicate that neither sodium hypochlorite nor ozone effectively inactivate T. gondii oocysts, even when used at high concentrations. PMID- 17918379 TI - Contribution of NADH dehydrogenase subunit I and cytochrome C oxidase subunit I sequences toward identifying a case of human coenuriasis in France. AB - Coenuriasis is a parasitic disease induced by larval taeniid tapeworms that is rarely observed in humans. In December 2005, a case was diagnosed in Nancy, France, after surgical excision of a cyst on a 24-yr-old woman returning from the Cote d'Ivoire. Morphological and epidemiological criteria suggested that the infection was due to Taenia serialis. Molecular analysis of NADH dehydrogenase subunit I (NDI) and cytochrome c oxidase subunit I (COI) sequences was also in favor of T. serialis identification, but the absence of available genetic data on T. brauni and T. glomeratus and the small number of published sequences for T. serialis and T. multiceps must be considered with caution. The NDI partial sequences presented more variations within species of Taenia than the COI sequences, which make them more useful targets for species identification and analysis of intraspecific polymorphisms. The present study points to the usefulness of molecular biology tools to help make up for the shortcomings of the commonplace parasitological diagnosis for coenuriasis. PMID- 17918380 TI - Checklist of helminth parasites of the cane toad Bufo marinus (Anura: Bufonidae) from Mexico. AB - Thirty-four adult cane toads Bufo marinus L. (12 males and 22 females) collected from 2 localities in Mexico (Cerro de Oro and Temascal Dams, Oaxaca) in September 2003 were examined for helminth parasites. In total, 14,749 helminths belonging to 14 taxa were collected. Included were 2 adult digeneans (Choledocystus hepaticus, Mesocoelium monas); 1 larval cestode (an unidentified pseudophyllidean); and 11 nematodes, including 3 species of larvae (Contracaecum sp., Physaloptera sp., Physocephalus sexalatus) and 8 species of adults (Aplectana itzocanensis, Cosmocerca sp., Cruzia morleyi, Ochoterenella digiticauda, Oswaldocruzia sp., Raillietnema sp., Rhabdias americanus, and Rhabdiasfuelleborni). Higher species richness was recorded in B. marinus from Cerro de Oro (12 taxa versus 9 in those from Temascal); hosts from both localities shared 7 taxa. There were 25 new locality records, and 2 taxa were registered in Mexico for the first time. To date, 112 helminth species have been recorded parasitizing B. marinus along its native and introduced range of distribution, with 40.5% of them reported from Mexico. PMID- 17918381 TI - Parasitic helminths of free-ranging mink (Neovison vison mink) from southern Florida. AB - Five free-ranging mink, Neovison vison mink, from in or near the Fakahatchee Strand Preserve State Park, Collier County, Florida (26 degrees 00'N, 81 degrees 25'W), were examined for parasitic helminths. Nine species of helminths were identified (2 trematodes, 5 nematodes, and 2 acanthocephalans). The most prevalent parasites were Molineus patens (4 of 5 mink), mean intensity 173 (range, 12-342); Strongyloides sp. (4 of 5), mean intensity 48 (range, 1-170); Macracanthorhynchus ingens (immature) (4 of 5), mean intensity 2.5 (range, 1-4); and Alaria mustelae (3 of 5), mean intensity 59.3 (range, 11-127). Polymorphus brevis and Dirofilaria lutrae are reported from the mink for the first time. PMID- 17918382 TI - Viability of the encysted metacercariae of Echinostoma caproni judged by light microscopy versus chemical excystation. AB - The purpose of this study was to determine the viability of encysted metacercariae of Echinostoma caproni stored in Locke's solution 1:1 at 4 C for 1 24 wk. Viability was judged by light microscopy (LM) based on morphological characteristics of the encysted metacercariae versus chemical excystation of the cysts in a trypsin-bile salts excystation medium. The percent viability was very similar under both methods of assessment at 4, 8, and 16 wk poststorage. At 1 and 24 wk poststorage, viability was found to be about 2x greater based on excystation than using LM. We concluded that LM alone underestimated the viability of cysts and that determination of cyst viability was more accurate under assessment by chemical excystation. PMID- 17918384 TI - Seroprevalence of Toxoplasma gondii in pigs from Vietnam. AB - Pigs are considered an important source of Toxoplasma gondii infection for humans. Antibodies to T. gondii were determined in serum samples from 587 pigs from Vietnam using the modified agglutination test (MAT) and found in 160 of 587 (27.2%) pigs, with MAT titers of 1:25 in 32 pigs, 1:50 in 34 pigs, 1:100 in 33 pigs, 1:200 in 24 pigs, 1:400 in 21 pigs, 1:800 in 14 pigs, and 1:3,200 in 2 pigs. Antibodies (MAT 1:20 or higher) were found in 75 of 325 (23%) finishers, 63 of 207 (32.3%) sows, and 22 of 55 (40%) boars. Results indicate high prevalence of T. gondii infection in pigs in Vietnam. This is the first report of prevalence of T. gondii in pigs from Vietnam. PMID- 17918383 TI - Coprology of Panthera tigris altaica and Felis bengalensis euptilurus from the Russian Far East. AB - Fecal samples from the Siberian tiger (Panthera tigris altaica) and the Amur cat (Felis bengalensis euptilurus) from Far Eastern Russia, were examined for parasites. A natural sedimentation methodology was used and a complete examination of all the sediment was performed. This fecal investigation allowed us to isolate and identify several developmental stages of gastrointestinal, hepatic, and respiratory parasites. Five parasites were found from P. t. altaica: 11 trematodes (Platynosomum fastosum) and 4 nematodes (Strongyloides sp., Ancylostomatidae, Toxascaris leonina, and Toxocara cati). Five parasites were found from F. b. euptilurus: 1 cestode (Diplopylidium sp.) and 4 nematodes (Trichuris sp., Ancylostomatidae, Toxascaris leonina, and Aelurostrongylus abstrusus). In addition, trophozoites of the amoeba Acanthamoeba sp. were detected in tiger feces. PMID- 17918385 TI - Seroprevalence of Toxoplasma gondii in cats from St. Kitts, West Indies. AB - The prevalence of antibodies to Toxoplasma gondii was determined in sera from 106 domestic cats from St. Kitts, West Indies. Using a modified agglutination test, antibodies to this parasite were found in 90 (84.9%) of the cats, with titers of 1:20 in 23 cats, 1:40 in 34 cats, 1:80 in 18 cats, 1:160 in 2 cats, 1:320 in 1 cat, and 1:1,280 or higher in 11 cats. This is the first report of the prevalence of T. gondii infections in cats on St. Kitts and suggests widespread contamination of the environment with oocysts. PMID- 17918386 TI - Bartonella and Rickettsia from fleas (Siphonaptera: Ceratophyllidae) of prairie dogs (Cynomys spp.) from the western United States. AB - Fleas of prairie dogs have been implicated in the transmission of Bartonella spp. We used PCR to test DNA extracts from 47 fleas of prairie dogs from 6 states. We amplified DNA from 5 unique genotypes of Bartonella spp. and 1 Rickettsia sp. from 12 fleas collected in North Dakota, Oklahoma, Texas, and Wyoming. Sequences from the Bartonella spp. were similar, but not identical, to those from prairie dogs and their fleas in Colorado. PMID- 17918387 TI - Prevalence of antibodies to Leishmania infantum and Trypanosoma cruzi in wild canids from South Carolina. AB - Wild canids are reservoir hosts for Leishmania infantum and Trypanosoma cruzi. The present study examined the prevalence of antibodies to these zoonotic parasites in a population of wild canids from a nonagricultural setting in South Carolina. Sera from 26 gray foxes (Urocyon cinereoargenteus) and 2 coyotes (Canis latrans) were examined for antibodies to L. infantum and T. cruzi using the indirect immunofluorescent antibody test and commercially available parasite specific immunochromatigraphic strip assays. Antibodies to L. infantum were not detected by either assay in gray foxes or coyotes. Two (8%) of 26 gray foxes were positive in both the T. cruzi immunofluorescent antibody and strip assays. Antibodies to T. cruzi were not detected in coyotes. Results from this study indicate that wild canids are exposed to T. cruzi, but not L. infantum. in this geographic region. PMID- 17918388 TI - Distribution pattern of phthirapterans infesting certain common Indian birds. AB - The prevalence and frequency distribution patterns of 10 phthirapteran species infesting house sparrows, Indian parakeets, common mynas, and white breasted kingfishers were recorded in the district of Rampur, India, during 2004-05. The sample mean abundances, mean intensities, range of infestations, variance to mean ratios, values of the exponent of the negative binomial distribution, and the indices of discrepancy were also computed. Frequency distribution patterns of all phthirapteran species were skewed, but the observed frequencies did not correspond to the negative binomial distribution. Thus, adult-nymph ratios varied in different species from 1:0.53 to 1:1.25. Sex ratios of different phthirapteran species ranged from 1:1.10 to 1:1.65 and were female biased. PMID- 17918389 TI - Feline cuterebrosis caused by a lagomorph-infesting Cuterebra spp. larva. AB - Native species of rodents and lagomorphs in the Americas are the typical hosts of Cuterebra spp. larvae. Although these bot flies are relatively host specific, they occasionally parasitize other native and introduced mammals (including domestic animals and humans), an affliction termed cuterebrosis. Cuterebra spp. larvae generally cause benign, subcutaneous lesions (warbles), but when infesting domestic cats, they can invade the eyes, respiratory tract, and cerebral tissues, causing severe, and in some cases fatal, injury. Despite more than 2 dozen published reports of feline cuterebrosis, the type (rodent- or lagomorph infesting) or species parasitizing domestic cats has rarely been determined. Here, I identify a larva removed from a kitten in southern Wisconsin as belonging to a lagomorph-infesting Cuterebra species, most likely C. abdominalis, based especially on features of the cuticular platelets covering its exterior, and its geographic location. This seems to be only the third substantiated report of feline cuterebrosis in more than 50 yr in which a larva has been identified beyond "Cuterebra spp." In each case, lagomorph-infesting species were involved, suggesting that domestic cats may not be susceptible to rodent-infesting Cuterebra species. However, because these studies are limited in number and geographic area, additional research is required to establish the spectrum of Cuterebra species involved in feline cuterebrosis. PMID- 17918390 TI - Molecular identification of two strains of third-stage larvae of Contracaecum rudolphii sensu lato (Nematoda: Anisakidae) from fish in Poland. AB - Contracaecum sp. larvae (L3) from fish were identified using nucleotide sequences of the internal transcribed spacers ITS-1 and ITS-2 of the ribosomal DNA. The nematode larvae originated from fish in a freshwater situation (crucian carp Carassius carassius, from Selment Wielki Lake in Mazury, northeastern Poland) and a brackish-water region (Caspian round goby Neogobius melanostomus from the Baltic Sea, Gdafisk Bay at the Polish coast). Two strains (Contracaecum rudolphii A and B) of Contracaecum rudolphii senso lato, a parasite common at the adult stage in fish-eating birds, were identified. In fish from the freshwater site, only the strain temporarily designated C. rudolphii B was identified; in the brackish-water region, both strains were found, suggesting that fish serve as paratenic host for both genotypes. Contracaecum rudolphii sensu lato has been recorded in several species of fish-eating birds in Poland, particularly in the great cormorant, Phalacrocorax carbo, in which the abundance is highest. The results, although based on a restricted number of larvae, suggest that the life cycles of both genotypes can be completed in the Polish region and that at least one of them, C. rudolphii B, can develop both in fresh and brackish water. PMID- 17918391 TI - Genetic and immunological characterization of the 14-3-3xi molecule from Schistosoma bovis. AB - Currently available candidate vaccines against schistosomiasis elicit only partial protection. In addition, the type of immune response that could lead to the highest level of protection against schistosomes has not yet been described. Thus, efforts should be made in both the identification of novel proteins essential for the parasite cycle and in the modulation of immune responses against these novel candidates through the combined use of immunomodulatory molecules. Several parasites have 14-3-3 proteins, and these proteins are known to play a key role in parasite biology. In the present work, we report the isolation and characterization of a new 14-3-3 gene from Schistosoma bovis and offer new information regarding the genetic structure of the gene. In addition, we have produced the corresponding recombinant protein. Finally, we describe the immune responses elicited by this protein when combined with 4 different immunomodulators in immunized mice. PMID- 17918392 TI - From the roots of parasitology: Hippocrates' first scientific observations in helminthology. AB - Although knowledge of intestinal parasites predates Hippocrates, the Hippocratic Corpus provides the first scientific observations about the clinical perception and treatment of helminthic diseases. These observations follow the scientific principles of Hippocrates, the father of modern medicine, who relied on knowledge and observation. This article is based on a systematic study of the Hippocratic texts, and presents observations on diseases caused by intestinal parasites with respect to regularity of appearance, patient age, symptoms, and treatment. Three types of helminths are described: "helmins strongyle" (roundworm), "helmins plateia" (flatworm), and "ascaris" (which corresponds to Enterobius vermicularis). Helminthic diseases primarily appear during childhood, well after teething. The described systemic symptoms include weakness, sickness, discomfort, tiredness, anorexia, and emotional instability; gastrointestinal symptoms include change in bowel movements, vomiting, and colic pain in the epigastrium. We identified several accounts of cases of helminthic diseases in the Hippocratic texts. Of particular interest are the descriptions of a helminth emerging from a fistula in the navel region and the surgical treatment of helminthic diseases, reinforced by being described on a dedicatory inscription at the Asclepion in Epidaurus. We finally encountered the use of powerful purgatives as antihelminthics, which have been widely used, even into the 21st century. PMID- 17918393 TI - Impact of curve construction and community dynamics on the species-time relationship. AB - The species-time relationship (STR) describes how the species richness of a community increases with the time span over which the community is observed. This temporal scaling provides insight into theoretical questions on species diversity patterns as well as applied questions on the appropriate time scale for biodiversity assessments. To better understand STRs, we discuss the methods used to construct STRs in the literature and derive the impact of curve construction on STR properties. Using vegetation data from Mount St. Helens, Washington, USA, we illustrate the sensitivity of the STR to construction under colonization dominated dynamics. This study highlights the importance of considering the type of STR when interpreting, comparing, and applying STRs, particularly in disturbed or successional systems. PMID- 17918394 TI - The role of environmental and spatial factors for the composition of aquatic bacterial communities. AB - This study investigates the importance of local vs. spatial factors on bacterial community composition of 35 rock pools at the Baltic Sea coast. The pools were located in five distinct spatial clusters over a total scale of <500 m and differed widely in terms of water chemistry. To determine the fractions of the variance in bacterial community composition (BCC) between rock pools that are explained by local environmental vs. spatial factors, a variance partitioning procedure using partial canonical correspondence analysis was performed. Three environmental variables (salinity, chlorophyll a concentration, and water color) had a significant effect on BCC, irrespective of the spatial location of the pools. Vice versa, there was a significant effect of spatial factors on BCC irrespective of any of the environmental factors included in this study. Hence, the patchy spatial distribution of the pools was partly reflected in the composition of the bacterial communities in the pools, which might be caused by congruent colonization events of adjacent pools, such as simultaneous seaspray inputs or direct exchange of bacteria via connecting rivulets. This study shows that the composition of planktonic bacteria can show provincialism at small spatial scales, which is likely to be caused by environmental conditions as well as historical events. PMID- 17918395 TI - Environmental controls on the landscape-scale biogeography of stream bacterial communities. AB - We determined the biogeographical distributions of stream bacteria and the biogeochemical factors that best explained heterogeneity for 23 locations within the Hubbard Brook watershed, a 3000-ha forested watershed in New Hampshire, USA. Our goal was to assess the factor, or set of factors, responsible for generating the biogeographical patterns exhibited by microorganisms at the landscape scale. We used DNA fingerprinting to characterize bacteria inhabiting fine benthic organic matter (FBOM) because of their important influence on stream nutrient dynamics. Across the watershed, streams of similar pH had similar FBOM bacterial communities. Streamwater pH was the single variable most strongly correlated with the relative distance between communities (Spearman's p = 0.66, P < 0.001) although there were other contributing factors, including the quality of the fine benthic organic matter and the amount of dissolved organic carbon and nitrogen in the stream water (P < 0.05 for each). There was no evidence of an effect of geographic distance on bacterial community composition, suggesting that dispersal limitation has little influence on the observed biogeographical patterns in streams across this landscape. Cloning and sequencing of small-subunit rRNA genes confirmed the DNA fingerprinting results and revealed strong shifts among bacterial groups along the pH gradient. With an increase in streamwater pH, the abundance of acidobacteria in the FBOM bacterial community decreased (from 71% to 38%), and the abundance of proteobacteria increased (from 11% to 47%). Together these results suggest that microorganisms, like "macro"-organisms, do exhibit biogeographical patterns at the landscape scale and that these patterns may be predictable based on biogeochemical factors. PMID- 17918396 TI - Allochthonous subsidy of periodical cicadas affects the dynamics and stability of pond communities. AB - Periodical cicadas emerge from below ground every 13 or 17 years in North American forests, with individual broods representing the synchronous movement of trillions of individuals across geographic regions. Due to predator satiation, most individuals escape predation, die, and become deposited as detritus. Some of this emergent biomass falls into woodland aquatic habitats (small streams and woodland ponds) and serves as a high-quality allochthonous detritus pulse in early summer. We present results of a two-part study in which we (1) quantified deposition of Brood X periodical cicada detritus into woodland ponds and low order streams in southwestern Ohio, and (2) conducted an outdoor mesocosm experiment in which we examined the effects of deposition of different amounts of cicada detritus on food webs characteristic of forest ponds. In the mesocosm experiment, we manipulated the amount of cicada detritus input to examine if food web dynamics and stability varied with the magnitude of this allochthonous resource subsidy, as predicted by numerous theoretical models. Deposition data indicate that, during years of periodical cicada emergence, cicada carcasses can represent a sizable pulse of allochthonous detritus to forest aquatic ecosystems. In the mesocosm experiment, cicada carcass deposition rapidly affected food webs, leading to substantial increases in nutrients and organism biomass, with the magnitude of increase dependent upon the amount of cicada detritus. Deposition of cicada detritus impacted the stability of organism functional groups and populations by affecting the temporal variability and biomass minima. However, contrary to theory, stability measures were not consistently related to the size of the allochthonous pulse (i.e., the amount of cicada detritus). Our study underscores the need for theory to further explore consequences of pulsed allochthonous subsidies for food web stability. PMID- 17918397 TI - Introduced trout sever trophic connections in watersheds: consequences for a declining amphibian. AB - Trophic linkages between terrestrial and aquatic ecosystems are increasingly recognized as important yet poorly known features of food webs. Here we describe research to understand the dynamics of lake food webs in relation to a native riparian amphibian and its interaction with introduced trout. The mountain yellow legged frog Rana muscosa is endemic to alpine watersheds of the Sierra Nevada Mountains and the Transverse Ranges of California, but it has declined to a small fraction of its historical distribution and abundance. Although remaining frogs and introduced trout feed in different habitats of alpine lakes, our stable isotope analyses clearly show that the same resource base of benthic invertebrates sustains their growth. During one period, insect emergence from naturally fishless lakes was nearly 20-fold higher compared to adjacent lakes with trout, showing that fish reduce availability of aquatic prey to amphibious and terrestrial consumers. Although trout cannot prey on adult frogs due to gape limitation, foraging post-metamorphic frogs are 10 times more abundant in the absence of trout, suggesting an important role for competition for prey by trout in highly unproductive alpine watersheds. Most Sierran lakes contain fish, and those that do not are usually small isolated ponds; in our study, these two lake types supported the lowest densities of post-metamorphic frogs, and these frogs were less reliant on local, benthic sources of productivity. Since Rana muscosa was formerly the most abundant vertebrate in the Sierra Nevada, the reduction in energy flow from lake benthos to this consumer due to fish introductions may have had negative consequences for its numerous terrestrial predators, many of which have also declined. We suggest that disruptions of trophic connections between aquatic and terrestrial food webs are an important but poorly understood consequence of fish introduction to many thousands of montane lakes and streams worldwide and may contribute to declines of native consumers in riparian habitats. PMID- 17918398 TI - Plankton biodiversity along a gradient of productivity and its mediation by macrophytes. AB - We studied the effect of aquatic vegetation on the process of species sorting and community assembly of three functional groups of plankton organisms (phytoplankton, seston-feeding zooplankton, and substrate-dwelling zooplankton) along a primary productivity gradient. We performed an outdoor cattle tank experiment (n = 60) making an orthogonal combination of a primary productivity gradient (four nutrient addition levels: 0, 10, 100, and 1000 microg P/L; N/P ratio: 16) with a vegetation gradient (no macrophytes, artificial macrophytes, and real Elodea nuttallii). We used artificial plants to evaluate the mere effects of plant physical structure independently from other plant effects, such as competition for nutrients or allelopathy. The tanks were inoculated with species-rich mixtures of phytoplankton and zooplankton. Both productivity and macrophytes affected community structure and diversity of the three functional groups. Taxon richness declined with increasing plankton productivity in each functional group according to a nested subset pattern. We found no evidence for unimodal diversity-productivity relationships. The proportional abundance of Daphnia and of colonial Scenedesmus increased strongly with productivity. GLM analyses suggest that the decline in richness of seston feeders was due to competitive exclusion by Daphnia at high productivity. The decline in richness of phytoplankton was probably caused by high Daphnia grazing. However, partial analyses indicate that these explanations do not entirely explain the patterns. Possibly, environmental deterioration associated with high productivity (e.g., high pH) was also responsible for the observed richness decline. Macrophytes had positive effects on the taxon richness of all three functional plankton groups and interacted with the initial productivity gradient in determining their communities. Macrophytes affected the composition and diversity of the three functional groups both by their physical structure and through other mechanisms. Part of the macrophyte effect may be indirect via a reduction of phytoplankton production. Our results also indirectly suggest that the often reported unimodal relationship between primary productivity and diversity in nature may be partially mediated by the tendency of submerged macrophytes to be most abundant at intermediate productivity levels. PMID- 17918399 TI - Whole-community mutualism: associated invertebrates facilitate a dominant habitat forming seaweed. AB - Many habitat-forming, or foundation, species harbor diverse assemblages of associated taxa that benefit from the refuges from predators or harsh physical conditions that foundation species provide. Growing numbers of studies show how specific taxa associated with foundation species can benefit their hosts, but the aggregate effects of the entire community of associated species remain poorly understood. Here, we evaluate the role that a diverse assemblage of invertebrates plays in mediating the dominance of a foundation species, the green filamentous seaweed Cladophora columbiana Collins, in rocky intertidal habitats. Cladophora is a fast-growing seaweed with a high nitrogen demand, and we suggest that it persists in nutrient-limited high-intertidal pools because of local-scale nitrogen excretion by the invertebrate taxa living within its filaments. Removal of associated invertebrates resulted in a fourfold increase in the rate of water column nitrogen depletion by Cladophora, and ammonium concentrations inside Cladophora turfs with invertebrates present were seven times higher than in the adjacent tide-pool water. The ammonium excreted by invertebrate meiofauna far surpassed the nitrogen used by Cladophora, suggesting that all of Cladophora's nitrogen requirements could be met by the invertebrates associated with it. This study links host performance to the total aggregate biomass of mutualists rather than the particular traits of any one species, suggesting the potential for important feedbacks between individual hosts and the communities of associated species that they support. PMID- 17918400 TI - Predictability of reef fish recruitment in a highly variable nursery habitat. AB - There has been a lengthy debate on whether the abundance of adult reef fishes depends on prerecruitment or postrecruitment processes; however, we still do not have the ability to predict the magnitude of local fish recruitment. Here we show that the success of the leopard grouper (Mycteroperca rosacea) recruitment in the Gulf of California, Mexico, is determined by the availability of nursery habitat, which in turn is strongly correlated to climate conditions. Observational and experimental studies showed that leopard grouper larvae recruit preferentially on shallow rocky bottoms with brown algal (Sargassum spp.) beds, and that abundance of recruits is determined by the availability of Sargassum. The biomass of Sargassum decreases linearly with an increase in the Multivariate El Nino Southern Oscillation (ENSO) Index (MEI; an index positively correlated with water temperature and negatively correlated with nutrient availability). We analyzed the relationship between the interannual variation of MEI and the recruitment of the leopard grouper using field estimates of abundance of juvenile groupers. Our results show that there is a nonlinear relationship between recruitment and the oceanographic climate, in that the density of recruits decreases exponentially with increasing MEI. The predictability of leopard grouper recruitment has important implications for fisheries management, since it could allow adaptive management without expensive stock assessment programs. PMID- 17918401 TI - Reproduction on the edge: large-scale patterns of individual performance in a marine invertebrate. AB - Reproductive output is a central attribute of life history, providing a measure of individual and population performance. The fields of ecology, biogeography, and evolutionary biology take disparate approaches in addressing spatial variation in reproduction, and thus we lack clear predictions for how reproductive output should vary geographically. We empirically investigate these contrasting theoretical approaches by determining geographic patterns in reproductive output for intertidal populations of the purple sea urchin, Strongylocentrotus purpuratus, at 15 sites spanning a large geographic distance (9 degrees span of latitude) from central California, USA, to Baja California, Mexico. Contrary to predictions from biogeography, some of the highest values of reproductive output are at sites near the species' southern range boundary. Additionally, sea urchins do not exhibit a latitudinal gradient in reproduction, but rather show considerable mesoscale variation in reproductive output. Spatial analyses reveal that this variation is correlated with coastal topographic features that are known to influence the pattern of nearshore nutrient flux and circulation. We hypothesize that urchins' reproductive output may be driven by the spatial distribution of their food supply, drift macroalgae, the abundance of which is influenced by both nutrient supply and alongshore transport processes that are coupled to topographic features. Large-scale studies such as this provide valuable insight into the causes of species' range limits, population connectivity, habitat reserve design, and forecasting the effects of climate change on species' distributions. PMID- 17918402 TI - New methods for quantifying the spatial storage effect: an illustration with desert annuals. AB - Recent theory supports the long-held proposition that coexistence is promoted by species-specific responses to a spatially varying environment. The underlying coexistence mechanism, the spatial storage effect, can be quantified by the covariance between response to the environment and competition. Here, "competition" is generalized to encompass similar processes such as facilitation and apparent competition. In the present study, we use a model field system of desert annual plants to demonstrate this method and to provide insight into the dynamics of the field system. Specifically, we use neighborhood competition experiments to quantify the spatial storage effect and compare it to the separate (but not mutually exclusive) process of neighborhood-scale resource partitioning. As our basic experimental design has been used frequently in community ecology, these methods can be applied to many existing data sets, as well as future field studies. PMID- 17918403 TI - Neighborhood and community interactions determine the spatial pattern of tropical tree seedling survival. AB - Factors affecting survival and recruitment of 3531 individually mapped seedlings of Myristicaceae were examined over three years in a highly diverse neotropical rain forest, at spatial scales of 1-9 m and 25 ha. We found convincing evidence of a community compensatory trend (CCT) in seedling survival (i.e., more abundant species had higher seedling mortality at the 25-ha scale), which suggests that density-dependent mortality may contribute to the spatial dynamics of seedling recruitment. Unlike previous studies, we demonstrate that the CCT was not caused by differences in microhabitat preferences or life history strategy among the study species. In local neighborhood analyses, the spatial autocorrelation of seedling survival was important at small spatial scales (1-5 m) but decayed rapidly with increasing distance. Relative seedling height had the greatest effect on seedling survival. Conspecific seedling density had a more negative effect on survival than heterospecific seedling density and was stronger and extended farther in rare species than in common species. Taken together, the CCT and neighborhood analyses suggest that seedling mortality is coupled more strongly to the landscape-scale abundance of conspecific large trees in common species and the local density of conspecific seedlings in rare species. We conclude that negative density dependence could promote species coexistence in this rain forest community but that the scale dependence of interactions differs between rare and common species. PMID- 17918404 TI - Mortality of large trees and lianas following experimental drought in an Amazon forest. AB - Severe drought episodes such as those associated with El Nino Southern Oscillation (ENSO) events influence large areas of tropical forest and may become more frequent in the future. One of the most important forest responses to severe drought is tree mortality, which alters forest structure, composition, carbon content, and flammability, and which varies widely. This study tests the hypothesis that tree mortality increases abruptly during drought episodes when plant-available soil water (PAW) declines below a critical minimum threshold. It also examines the effect of tree size, plant life form (palm, liana, tree) and potential canopy position (understory, midcanopy, overstory) on drought-induced plant mortality. A severe, four-year drought episode was simulated by excluding 60% of incoming throughfall during each wet season using plastic panels installed in the understory of a 1-ha forest treatment plot, while a 1-ha control plot received normal rainfall. After 3.2 years, the treatment resulted in a 38% increase in mortality rates across all stems >2 cm dbh. Mortality rates increased 4.5-fold among large trees (>30 cm dbh) and twofold among medium trees (10-30 cm dbh) in response to the treatment, whereas the smallest stems were less responsive. Recruitment rates did not compensate for the elevated mortality of larger-diameter stems in the treatment plot. Overall, lianas proved more susceptible to drought-induced mortality than trees or palms, and potential overstory tree species were more vulnerable than midcanopy and understory species. Large stems contributed to 90% of the pretreatment live aboveground biomass in both plots. Large-tree mortality resulting from the treatment generated 3.4 times more dead biomass than the control plot. The dramatic mortality response suggests significant, adverse impacts on the global carbon cycle if climatic changes follow current trends. PMID- 17918405 TI - Woody plant richness and NDVI response to drought events in Catalonian (northeastern Spain) forests. AB - The role of species diversity on ecosystem resistance in the face of strong environmental fluctuations has been addressed from both theoretical and experimental viewpoints to reveal a variety of positive and negative relationships. Here we explore empirically the relationship between the richness of forest woody species and canopy resistance to extreme drought episodes. We compare richness data from an extensive forest inventory to a temporal series of satellite imagery that estimated drought impact on forest canopy as NDVI (normalized difference vegetation index) anomalies of the dry summer in 2003 in relation to records of previous years. We considered five different types of forests that are representative of the main climatic and altitudinal gradients of the region, ranging from lowland Mediterranean to mountain boreal-temperate climates. The observed relationship differed among forest types and interacted with the climate, summarised by the Thorntwaite index. In Mediterranean Pinus halepensis forests, NDVI decreased during the drought. This decrease was stronger in forests with lower richness. In Mediterranean evergreen forests of Quercus ilex, drought did not result in an overall NDVI loss, but lower NDVI values were observed in drier localities with lower richness, and in more moist localities with higher number of species. In mountain Pinus sylvestris forests NDVI decreased, mostly due to the drought impact on drier localities, while no relation to species richness was observed. In moist Fagus sylvatica forests, NDVI only decreased in plots with high richness. No effect of drought was observed in the high mountain Pinus uncinata forests. Our results show that a shift on the diversity-stability relationship appears across the regional, climatic gradient. A positive relationship appears in drier localities, supporting a null model where the probability of finding a species able to cope with drier conditions increases with the number of species. However, in more moist localities we hypothesize that the proportion of drought-sensitive species would increase in richer localities, due to a higher likelihood of co-occurrence of species that share moist climatic requirements. The study points to the convenience of considering the causes of disturbance in relation to current environmental gradients and historical environmental constraints on the community. PMID- 17918406 TI - Process-based modeling of species' distributions: what limits temperate tree species' range boundaries? AB - Niche-based models are widely used to understand what environmental factors determine species' distributions, but they do not provide a clear framework to study the processes involved in defining species' ranges. Here we used a process based model to identify these processes and to assess the potential distribution of 17 North American boreal/temperate tree species. Using input of only climate and soil properties, the model reproduced the 17 species' distributions accurately. Our results allowed us to identify the climatic factors as well as the biological processes involved in limiting species' ranges. The model showed that climatic constraints limit species' distributions mainly through their impact on phenological processes, and secondarily through their impact on drought and frost mortality. The northern limit of species' ranges appears to be caused mainly by the inability to undergo full fruit ripening and/or flowering, while the southern limit is caused by the inability to flower or by frost injury to flowers. These findings about the ecological processes shaping tree species' distribution represent a crucial step toward obtaining a more complete picture of the potential impact of climate on species' ranges. PMID- 17918407 TI - Soil vs. canopy seed storage and plant species coexistence in species-rich Australian shrublands. AB - The fire-prone shrublands of southwestern Australia are renowned for their high plant species diversity and prominence of canopy seed storage (serotiny). We compared species richness, abundance, and life history attributes for soil and canopy seed banks in relation to extant vegetation among four sites with different substrate conditions and high species turnover (50-80%) to identify whether this unusual community-level organization of seed storage might contribute to maintenance of high species richness. Soil seed bank (SSB) densities were low to moderate (233-1435 seeds/m2) compared with densities for other Mediterranean-type vegetation and were lowest for sites with highest canopy seed bank (CSB) species richness and lowest nutrient availability, but not richness or abundance of resprouters. Annuals were infrequent in the lowest nutrient sites, but there was no evidence that small SSB size was due to low seed inputs or a trade-off between seed production/storage and seed size in response to low nutrient availability. Sorensen's similarity between SSB and extant vegetation was 26-43% but increased to 54-57% when the CSB was included, representing levels higher than reported for most other ecosystems. Resprouting species were well represented in both the SSB and CSB, and there was no evidence for lower seed production in resprouters than in non-sprouters overall. The SSB and CSB held no species in common and were characterized by markedly different seed dispersal attributes, with winged or small seeds in the CSB and seeds dispersed by ants, birds, and wind (though none with wings) in the SSB. There was no evidence of spatial differentiation in the distribution of seeds of SSB species between vegetated and open microsites that might facilitate species coexistence, but most woody non-sprouters showed aggregation at scales of 1-2 m, implying limited seed dispersal. High similarity between overall seed bank (SSB + CSB) and extant species composition, high number of resprouting species, and seed dispersal processes before (SSB) and after fire (CSB) leading to differential spatial aggregation of post-fire recruits from the two seed bank types may buffer species composition against rapid change and provide a mechanism for maintaining species coexistence at the local scale. PMID- 17918408 TI - Controls on vegetation structure in Southwestern ponderosa pine forests, 1941 and 2004. AB - Long-term studies can broaden our ecological understanding and are particularly important when examining contingent effects that involve changes to dominance by long-lived species. Such a change occurred during the last century in Southwestern (USA) ponderosa pine (Pinus ponderosa) forests. We used five livestock grazing exclosures established in 1912 to quantify vegetation structure in 1941 and 2004. Our objectives were to (1) assess the effects of historical livestock grazing on overstory structure and age distribution, (2) assess the effects of recent livestock grazing and overstory on understory vegetation, and (3) quantify and explain changes in understory vegetation between 1941 and 2004. In 1941, canopy cover of tree regeneration was significantly higher inside exclosures. In 2004, total tree canopy cover was twice as high, density was three times higher, trees were smaller, and total basal area was 40% higher inside exclosures. Understory species density, herbaceous plant density, and herbaceous cover were negatively correlated with overstory vegetation in both years. Most understory variables did not differ between grazing treatments in 1941 but were lower inside exclosures in 2004. Differences between grazing treatments disappeared once overstory effects were accounted for, indicating that they were due to the differential overstory response to historical livestock grazing practices. Between 1941 and 2004, species density declined by 34%, herbaceous plant density by 37%, shrub cover by 69%, total herbaceous cover by 59%, graminoid cover by 39%, and forb cover by 82%. However, these variables did not differ between grazing treatments or years once overstory effects were accounted for, indicating that the declines were driven by the increased dominance of the overstory during this period. Our results demonstrate that historical livestock grazing practices are an aspect of land-use history that can affect ecosystem development. Grazing history must be considered when extrapolating results from one site to another. In addition, the understory vegetation was more strongly controlled by the ponderosa pine overstory than by recent livestock grazing or by temporal dynamics, indicating that overstory effects must be accounted for when examining understory responses in this ecosystem. PMID- 17918409 TI - Moss species benefits from breakdown of cyclic rodent dynamics in boreal forests. AB - Bryophytes have increased in abundance in northern regions, and climate changes have been proposed to account for this change. However, changes in the population dynamics of microtine rodents may also contribute to changes in bryophyte abundance. New evidence indicates a tendency for microtine rodent population oscillations to change from periodicity of 3-5 years to become irregular or acyclic. The impact on ecosystem functioning is potentially great. We study the impact of variation in microtine rodent population characteristics, such as cycle length and amplitude, on the population dynamics of the boreal, clonal moss Hylocomium splendens. We use experimental and observational demographic data to construct 127 scenarios representing all combinations of disturbance type (gap formation and/or clipping), period (cyclic with 4, 6, 12, or 24 years between rodent peaks; or acyclic with constant or stochastically varying annual disturbance severity) and disturbance severity (fraction of individuals affected by disturbance in each year relative to the maximum disturbance carried out in the field experiment; seven levels). Population data collected in the field during 13 years were used as a baseline scenario. By subjecting all scenarios to stochastic matrix modeling, we demonstrate considerable impact of microtine rodent on the population dynamics of H. splendens, most notably when rodent populations fluctuate with short periods and high peak disturbance severities. Under the same average disturbance severity, H. splendens population growth rates are highest in acyclic scenarios and are progressively reduced with increasing peak disturbance severities (i.e., with increasing period). Stochastic elasticity analyses show that in less variable environments mature segment survival contributes more to the population growth rate, while in more variable environments the regeneration pathway (branching of older parts of the plant) plays a stronger role, inevitably leading to lower population fitness. Our results support the hypothesis that breakdown of cyclic rodent population dynamics accentuates increase in the abundance of H. splendens and other large bryophytes in boreal forests in Norway, observed empirically in recent years and primarily ascribed to climatic change. PMID- 17918410 TI - Interference competition between introduced black rats and endemic Galapagos rice rats. AB - Replicated field experiments were used to quantify and to describe the mechanism of competition between the introduced black rat Rattus rattus and the endemic Santiago rice rat Nesoryzomys swarthi on Santiago Island, Galapagos Islands, Ecuador. The removal of R. rattus significantly slowed the rate of seasonal population decline in N. swarthi. This effect was particularly evident for female, relative to male, N. swarthi and appeared to be driven solely by enhanced immigration; no other fitness or space use parameters were affected. The candidate hypotheses to explain the mechanism of competition were exploitation competition, interference by resource defense, and interference by aggressive encounter. To distinguish between hypotheses, we conducted a replicated resource supplementation experiment with patchy food, scattered food, and no food (control) treatments. The opportunistic R. rattus responded to the extra resources with increased adult immigration and juvenile recruitment, resulting in a significant abundance boost of sevenfold on patchy grids and fourfold on scattered grids. Females increased in body mass, and the breeding season was lengthened. In contrast, there was no change in the abundance of N. swarthi and no obvious benefit to reproduction. Instead, the costs of interference apparently outweighed the benefits of extra food: female N. swarthi increased in mass with supplementary food, but female (relative to male) immigration and residency were repressed on all supplemented areas. This response supported the hypothesis of interference by aggressive encounter, and we were able to rule out the alternative hypotheses. Although periodic population crashes of R. rattus on the arid north coast of Santiago may ameliorate its competitive impact, climate change may tip the balance. Control or eradication of R. rattus should improve future survival prospects for N. swarthi, but wildlife managers must be prepared for the potential eruption of the introduced house mouse Mus musculus, because this species experienced a release from interference competition and immigrated to removal areas. PMID- 17918411 TI - Extinction-effective population index: incorporating life-history variations in population viability analysis. AB - Viability status of populations is a commonly used measure for decision-making in the management of populations. One of the challenges faced by managers is the need to consistently allocate management effort among populations. This allocation should in part be based on comparison of extinction risks among populations. Unfortunately, common criteria that use minimum viable population size or count-based population viability analysis (PVA) often do not provide results that are comparable among populations, primarily because they lack consistency in determining population size measures and threshold levels of population size (e.g., minimum viable population size and quasi-extinction threshold). Here I introduce a new index called the "extinction-effective population index," which accounts for differential effects of demographic stochasticity among organisms with different life-history strategies and among individuals in different life stages. This index is expected to become a new way of determining minimum viable population size criteria and also complement the count-based PVA. The index accounts for the difference in life-history strategies of organisms, which are modeled using matrix population models. The extinction effective population index, sensitivity, and elasticity are demonstrated in three species of Pacific salmonids. The interpretation of the index is also provided by comparing them with existing demographic indices. Finally, a measure of life history-specific effect of demographic stochasticity is derived. PMID- 17918412 TI - Analyzing animal movements using Brownian bridges. AB - By studying animal movements, researchers can gain insight into many of the ecological characteristics and processes important for understanding population level dynamics. We developed a Brownian bridge movement model (BBMM) for estimating the expected movement path of an animal, using discrete location data obtained at relatively short time intervals. The BBMM is based on the properties of a conditional random walk between successive pairs of locations, dependent on the time between locations, the distance between locations, and the Brownian motion variance that is related to the animal's mobility. We describe two critical developments that enable widespread use of the BBMM, including a derivation of the model when location data are measured with error and a maximum likelihood approach for estimating the Brownian motion variance. After the BBMM is fitted to location data, an estimate of the animal's probability of occurrence can be generated for an area during the time of observation. To illustrate potential applications, we provide three examples: estimating animal home ranges, estimating animal migration routes, and evaluating the influence of fine-scale resource selection on animal movement patterns. PMID- 17918413 TI - Estimating trend precision and power to detect trends across grouped count data. AB - Ecologists commonly use grouped or clustered count data to estimate temporal trends in counts, abundance indices, or abundance. For example, the U.S. Breeding Bird Survey data represent multiple counts of birds from within each of multiple, spatially defined routes. Despite a reliance on grouped counts, analytical methods for prospectively estimating precision of trend estimates or statistical power to detect trends that explicitly acknowledge the characteristics of grouped count data are undescribed. These characteristics include the fact that the sampling variance is an increasing function of the mean, and that sampling and group-level variance estimates are generally estimated on different scales (the sampling and log scales, respectively). We address these issues for repeated sampling of a single population using an analytical approach that has the flavor of a generalized linear mixed model, specifically that of a negative binomial distributed count variable with random group effects. The count mean, including grand intercept, trend, and random group effects, is modeled linearly on the log scale, while sampling variance of the mean is estimated on the log scale via the delta method. Results compared favorably with those derived using Monte Carlo simulations. For example, at trend = 5% per temporal unit, differences in standard errors and in power were modest relative to those estimated by simulation (< or = /11/% and < or = /16/%, respectively), with relative differences among power estimates decreasing to < or = /7/% when power estimated by simulations was > or = 0.50. Similar findings were obtained using data from nine surveys of fingernail clams in the Mississippi River. The proposed method is suggested (1) where simulations are not practical and relative precision or power is desired, or (2) when multiple precision or power calculations are required and where the accuracy of a fraction of those calculations will be confirmed using simulations. PMID- 17918414 TI - Blue Tits use fledgling quantity and quality as public information in breeding site choice. AB - Public information (PI), which is the information that can be derived from the behavior and performance of conspecifics, has been demonstrated to be used in many fitness-enhancing decisions. In the context of breeding habitat choice, PI use has been called "habitat copying." We experimentally tested the existence of habitat copying in the Blue Tit (Cyanistes caeruleus), a nonmigratory, short lived hole-nesting bird. We manipulated the mean number of fledglings raised locally (quantity) and their condition (quality) as components of PI by transferring nestlings from Decreased (D) patches to Increased (I) patches. Our manipulations caused a negative relationship between fledgling quantity and quality that does not exist naturally: I patches had a higher number of fledglings that were in poorer condition, whereas D patches had a lower number in better condition. Control (C) patches, whether manipulated or not, had intermediate levels in terms of fledgling quantity and quality. Adult emigration the following year was higher from D than from C or I patches. Similarly, adult dispersal distance decreased for individuals coming from D to C to I patches. This suggests that resident breeders rely mainly on fledgling quantity to make emigration decisions. Emigration patterns of juveniles did not vary in relation to our patch manipulation. Immigration rates were higher and similar in I and D patches than in C patches. Hence, immigrant Blue Tits seem to rely on one of the manipulated components of PI and are insensitive to the discrepancy between fledgling quantity and quality. This shows that even nonmigratory species, such as Blue Tits, may use PI in their dispersal decisions but weigh its components differently for emigration and immigration. Differences among species in the importance of PI in breeding habitat choices may be explained by differences in life histories. PMID- 17918415 TI - Insects mediate the effects of propagule supply and resource availability on a plant invasion. AB - Invasive species are a global threat to biodiversity and the functioning of natural ecosystems. Here, we report on a two-year experiment aimed at elucidating the combined and relative effects of three key controls on plant invasions: propagule supply, soil nitrogen (N) availability, and herbivory by native insects. We focus on the exotic species Lespedeza cuneata, a Rank 1 invasive species. Propagule supply and soil N-availability interacted to control the density and foliar cover of L. cuneata. In low N plots, density and foliar cover of L. cuneata were higher in the propagule addition plots than in the plots to which propagules were not added. Surprisingly, this interaction was significant only when the abundance of herbivores was experimentally reduced. This experiment provides evidence that native insect herbivores mediate the interactive effects of propagule supply and resources on invasion by a widespread invasive plant species. PMID- 17918416 TI - Invading parasitoids suffer no Allee effect: a manipulative field experiment. AB - One frequent explanation for the failure of biological invasions is the Allee effect: due to positive density dependence, initially small invading populations may fail to establish and spread. Populations released for biological control are similar to fortuitous invading populations and may therefore suffer from Allee effects. However, unlike fortuitous invasions, biological control allows the experimental manipulation of initial population size and, thus, offers a unique opportunity to test for the occurrence of Allee effects. We manipulated the initial size of 45 populations of a parasitoid wasp introduced for the biological control of a phytophagous insect and followed the population dynamics of both parasitoids and hosts during three years. Our results suggest an absence of Allee effects but clear negative density dependence instead: (1) the probability of establishment after three years was not affected by initial population size; (2) net reproductive rate was highest at low parasitoid density and high host density; (3) the sex ratio, reflecting the proportion of virgin females, did not increase at low density, suggesting that low densities did not impede mate finding; (4) the depression of host populations did not depend upon the number of parasitoids introduced. This is, to our knowledge, the first experimental test of the Allee effect in an invading parasitoid. It leads us to propose that a number of behavioral and life-history features of many parasitoids could protect them from Allee effects. PMID- 17918417 TI - Facilitation across stress gradients: the importance of local adaptation. AB - While there is some information on genetic variation in response to competition in plants, we know nothing about intraspecific variation in facilitation. Previous studies suggest that facilitation should increase fitness in stressful environments. However, whether a plant experiences an environment as stressful may depend on prior adaptive responses to stressors at a site. Local adaptation to stress at a site may reduce the likelihood of facilitation. Seeds of Plantago erecta from stressful (serpentine soil) and non-stressful (non-serpentine soil) edaphic environments were reciprocally planted into these two soil types. Although competition did not differ significantly among seed sources, there was evidence for a local adaptation effect on facilitation. Non-serpentine seeds planted into serpentine soil exhibited greater individual plant biomass at higher densities. The interaction between population source and growth environment indicates a role for evolutionary processes such as local adaptation in the expression of facilitation in plants. PMID- 17918418 TI - Retrograde amnesia for extinction: similarities with amnesia for original acquisition memories. AB - Two experiments were conducted using rats to determine whether extinction is susceptible to a traditional amnestic agent (i.e., hypothermia) and to examine whether amnesia for extinction follows the same characteristics as those that occur with original memories. In Experiment 1, rats received hypothermia immediately, 60 min, or 120 min after extinction. When tested, the subjects cooled shortly after extinction showed little memory of the extinction training. This amnesia for extinction disappeared with longer postextinction delays, demonstrating a temporal gradient. Experiment 2 replicated the basic finding and demonstrated that an amnestic-extinguished memory could be recovered by reexposing the subjects to the amnestic agent and that the recovered extinction memory did not persist. These findings provide more evidence that extinction is a form of new learning and are consistent with retrograde amnesia research for original memories showing a temporal gradient and alleviation of retrograde amnesia by reexposure to the amnestic agent. PMID- 17918419 TI - Divided attention and the matching law: sample duration affects sensitivity to reinforcement allocation. AB - Previously, we have shown that changes in pigeons' divided attention performance resulting from changes in relative reinforcement are well described by the generalized matching law. In the present experiment, we examined whether sensitivity of performance to variations in relative reinforcement would be dependent upon sample duration. Pigeons responded on a delayed matching-to-sample procedure with compound samples (color + line orientation) and element comparison stimuli (two colors or two line orientations). Relative reinforcement for accurate matches on the two types of comparison trials varied across conditions. Sample duration was short (i.e., 0.75 sec) for half of the trials in a session and longer (i.e., 2.25 sec) for the other half. Sensitivity of accuracy to changes in relative reinforcement was greater with the longer sample than with the shorter sample, suggesting that differential reinforcement alters the allocation of attending to the elements of compound stimuli. Continued examination of the applicability of well-established theories of goal-directed behavior to the allocation of attention may provide further insights into what is variously referred to as goal-directed, voluntary, endogenous, or top-down control of attention. PMID- 17918420 TI - Discrimination blocking: acquisition versus performance deficits in human contingency learning. AB - We compared acquisition and performance accounts of human contingency learning. After solving a discrimination in Phase 1, in which Cue A predicted the occurrence of the outcome and Cue B predicted its nonoccurrence (A+/B-), a new discrimination (X+/Y-) was superimposed in Phase 2 (AX+/BY-). The participants were finally trained in Phase 3 with the added discrimination, which either maintained the same contingencies as those in Phase 2 (X+/Y-; Experiment 1) or reversed the contingencies (X-/Y+; Experiment 2). According to competitive learning theories, there should be no learning of the added discrimination in Phase 2, so that no advantage or disadvantage for this discrimination should be observed in Phase 3. In contrast, performance theories, such as the comparator hypothesis, contend that learning of the added discrimination in Phase 2 should proceed normally; so, in Phase 3, an advantage for the added discrimination should be observed in Experiment 1, but a disadvantage should be observed in Experiment 2. Our participants learned about the added discrimination and generally showed the effects predicted by the comparator hypothesis. PMID- 17918421 TI - Temporal discrimination of alternate days in rats. AB - The objective was to determine whether rats could learn to time a 48-h interval. Rats (n = 6) were continuously housed in operant chambers in constant darkness. The feeding cycle consisted of unlimited access to food for 6 h, followed by 42 h without access to food (i.e., meals were available on alternate days, contingent on breaking a photobeam in the food trough). Head entries into the trough increased as a function of time prior to the meal; this increase was higher, relative to the increase that occurred at the same time of day on alternate (i.e., nonfood) days. These data suggest that rats discriminated alternate days. Next, two meals were omitted to dissociate mechanisms of a self-sustained endogenous rhythm, interval timing, and alternation. Response rate increased periodically every 24 h, which suggests that the rats anticipated alternate days by discriminating the status of the previous day as a meal or a nonmeal day. PMID- 17918422 TI - Rats' choices between one and two delayed reinforcers. AB - Rats chose between alternatives that differed in the number of reinforcers and in the delay to each reinforcer. A left leverpress led to two reinforcers, each delivered after a fixed delay. A right leverpress led to one reinforcer after an adjusting delay. The adjusting delay was increased or decreased many times in a session, depending on the rat's choices, in order to estimate an indifference point--a delay at which the two alternatives were chosen about equally often. Both the number of reinforcers and their individual delays affected the indifference points. The overall pattern of results was well described by the hyperbolic-decay model, which states that each additional reinforcer delivered by an alternative increases preference for that alternative but that a reinforcer's effect is inversely related to its delay. Two other possible delay-discounting equations, an exponential equation and a reciprocal equation, did not produce satisfactory predictions for these data. Adding an additional free parameter to the hyperbolic equation as an exponent for delay did not appreciably improve the predictions, suggesting that raising delay to some power other than 1.0 was unnecessary. The results were qualitatively similar to those from a previous experiment with pigeons, but quantitative differences suggested that the rates of delay discounting were several times slower for rats than for pigeons. PMID- 17918423 TI - Remembering as discrimination in delayed matching to sample: discriminability and bias. AB - Task difficulty in delayed matching-to-sample tasks (DMTS) is increased by increasing the length of a retention interval. When tasks become more difficult, choice behavior becomes more susceptible to bias produced by unequal reinforcer ratios. Delaying reinforcement from choice behavior also increases both task difficulty and the biasing effect of unequal reinforcer probability. Six pigeons completed nine DMTS conditions with retention intervals of 0, 2, 4, 6, and 8 sec, in which reinforcer delays of 0, 2, and 4 sec were combined with ratios of reinforcer probabilities of .5/.5, .2/.8, and .8/.2 for correct red and green responses. Discriminability (log d) decreased with both increasing retention interval duration and increasing reinforcer delay. Sensitivity to reinforcement, the tendency for ratios of choice responses to follow unequal reinforcer probabilities, also increased as a function of both increasing retention interval and increasing reinforcer delay. The result is consistent with the view that remembering in DMTS tasks is a discriminated operant in which increasing task difficulty increases sensitivity to reinforcement. PMID- 17918424 TI - Pigeons learn to answer the question "where did you just peck?" and can report peck location when unexpectedly asked. AB - If pigeons are trained on matching-to-sample with differential responding required to the two samples, there is evidence that the differential responding can control comparison choice. We asked whether similar responding required at two different locations could also serve as the basis for comparison choice. Pigeons were pretrained to report the location that they had pecked. To reduce the likelihood that they could use the presence of differential proprioceptive cues at the time of their report, a common response was required between the location response and the comparison choice. They were then given experience with a conditional discrimination in which location of the comparison response varied randomly and was incidental to the choice of comparison. On test trials, after the pigeons had made their comparison choice, they showed a significant tendency to choose the appropriate test comparison when they were unexpectedly asked to report the location of their previous pecking response. These results have implications for the demonstration of episodic-like memory in pigeons because they suggest that pigeons have the capacity to recall, unexpectedly, specific details about their past experiences. PMID- 17918425 TI - [Rapid propagation of Echinacea angustifolia]. AB - The embryo of Echinacea angustifolia was used for explants to induce shoots and callus on MS medium. The plantlets could produce from embryo directly and the shoots could be induced to form fasciculate buds through the callus differentiation. The green plantlets from embryo or fasciculated buds were used for rooting inducement. The experiment showed that the embryo pre-cultured at 4 degrees C and in dark for 7 days can germinate easier than at other cultural conditions; Treatment with low temperature and dark were favorable for growth of radicle and plumule and development of green plantlets; A single shoot could proliferate rapidly to form fasciculated buds by means of callus induction and differentiation; the optimal medium for growth and proliferate were MS + BA 1.0 mg/L + NAA 0.5 mg/L. With this method, the proliferous coefficient should be achieved 3.05. PMID- 17918426 TI - [Study on immunological effect of housefly protein in mice]. AB - OBJECTIVE: To study the immunological effect of the housefly protein in mice. METHODS: The housefly protein was given orally in normal and Cyclophosphamide (CY) immunosuppressive mice, and then the indexes of the clearance rate of carbon particles and the serum hemolysin against SRBC were detected. RESULTS: The housefly protein could increase HC50 significantly in the normal mice, but could not in the CY immunosppressive mice. It could increase phagocytic index significantly in the normal mice, but could not in the CY immunosuppressive mice. CONCLUSION: The results suggest that the housefly protein can promote humoral immunity functions of normal mice. It has distinct preventive and cure effect on the CY immunosuppressive. PMID- 17918427 TI - [Study on the chemical changes of whole constituents in the preparing process of kansui root]. AB - OBJECTIVE: To study the chemical changes of whole constituents in the preparing process of kansui root in order to uncover the mechanism of detoxication in preparing process. METHODS: The raw and prepared kansui were extracted with water and methanol and analyzed with HPLC respectively. RESULTS: Seven constituents disappeared in the prepared kansui and four new consitituents were produced in the water extracts. The concentration of four other constituents decreased. In the methoanol extract, two constituents disappeared and one constituent produced during preparing process. The concentration of six other constituents increased in the methanol extracts. CONCLUSION: The mechanism of detoxication of kansui root preparing process is may be that the toxic constituents decomposing or water solubility reducing. PMID- 17918428 TI - [Identification of Ligustrum lucidum Ait. by X-ray diffraction fourier pattern method]. AB - OBJECTIVE: To set up a new identification and analysis method of Ligustrum lucidum Ait. METHODS: To get the extracts of Ligustrum lucidum Ait. using ethanol and chloroform ether as solvent and the extracts were identified by X-ray diffraction Fourier fingerprint spectra. RESULTS: Experiments and analysis were carried out on four samples. The standard X-ray diffraction Fourier fingerprint spectra and characteristic diffraction peaks were obtained. There were some differences among the spectra of the extracts, but the characteristic diffraction peaks were obvious. CONCLUSION: The experimental result indicated that X-ray diffraction Fourier fingerprint spectra can be used to identify and analyze Chinese traditional herb Ligustrum lucidum Ait. PMID- 17918429 TI - [Quality analysis of Radix Salviae Miltiorrhizae from different habitats by HPLC and TLC]. AB - OBJECTIVE: To analyze and compare the composition of Radix Salviae Miltiorrhizae from different habitats by HPLC and TLC. METHODS: The fingerprints of Radix Salviae Miltiorrhizae were detected by HPLC and TLC. RESULTS: Radix Salviae Miltiorrhiza collected from different habitats showed diverse fingerprints of component. The content of tanshinone II A of Radix Salviae Miltiorrhizae in Shandong was the highest, which is 3.5 times higher than Shangluo of Shanxi province. CONCLUSION: The character of two analytical mehods with fingerprint of component is significant, simplicity, and reproducibility is great. The methods can be used to select Radix Salviae Miltiorrhizae for clinical and resarch institution. PMID- 17918430 TI - [Studies on chemical constituents of Lobaria kurokawae yoshim]. AB - Six compounds have been isolated from the lichen plant Lobaria kurokawae and their structures were determined as ergosterol (1) , stictan-3beta, 22alpha-diol (2), retigeric acid B (5), retigeric acid A (6), ethyl orsellinate (3)and 3-O methyl-1,2: 5,6-bis-O-(1 -mehtylethylidene)-d-chiro-inostiol (4) on the basis of spectral data(MS, 1H-NMR, 13C-NMR and 2D-NMR). Compounds 1-4 were isolated from this plant for the first time. PMID- 17918431 TI - [Studies on chemical constituents of marine bryozoan Bugula neritina L]. AB - Seven compounds were isolated from the marine bryozoan Bugula neritina L. Their chemical structures were identified by NMR and MS spectroscopies as follows: cholesterol (I), cholest-4-en-3-one (II), cholesteryl myristate (III), 3beta,5alpha,9alpha-trihydrox-y-(22E,24R)-ergosta-,22-dien-6-one (IV), 3beta,5alpha,6beta-trihydroxy-(22E,24R)-ergosta-7,22-dien (V), uracil (VI), thymine (VII). Compounds II-VII were isolated from Bugula neritina L. for the first time. PMID- 17918432 TI - [Studies on flavonoids of Eupatorium odoratum L]. AB - OBJECTIVE: To investigate the flavonoid constituents from aerial parts of Eupatorium odoratum L. systematically. METHODS: The constituents of the EtoAc soluble portions of the 95% ethanol extraction were isolated and purified by silica gel and sephadex LH-20 chromatography. Their structures were elucidated on the basis of physicochemical properties and spectral analysis. RESULTS: Eight compounds were isolated and identified as kaempferol-3-methoxy (I), rhamnetin (II), tamarixetin (III), quercetin (IV), kaempferol (V), apigenin (VI), luteolin (VII), dihydrokaempferide (VIII). CONCLUSION: Compound I , II and VI were isolated for the first time from genus Eupatorium, and VIII which is rare flavonoid was obtainted for the first time from family Compositae. PMID- 17918433 TI - [Rotenoids from Derris trifoliata]. AB - Five rotenoids named dehydrodeguelin, dehydrorotenone, 12a-hydroxy rotenone, tephrosin and 12alphabeta-hydroxyrot-2'-enonic acid were isolated from aerial parts of Derris trifoliata. All these compounds were isolated from this plant for the first time. PMID- 17918434 TI - [Studies on the chemical constituents of Urtica dioica L. grown in Tibet Autonomous Region]. AB - Studies on the whole herb of Urtica dioica L. grown in Nyingchi area, China's Tibet Autonomous Region, resulted in the isolated of nine compounds: beta sitosterol, trans-ferulic acid, dotriacotane, erucic acid, ursolic acid, scopoletin, rutin, quercetin and p-hydroxylbenzalcohol. Dotriacotane, erucic acid, scopoletin, rutin and p-hydroxylbenzalcohol were obtained from Urtica L. for the first time. Their structures were confirmed by modem spectral analysis (NMR, MS, etc). PMID- 17918435 TI - [Analysis of lipid compounds of high-yielded rhizoma pinelliae growing in the west of Hubei province by gas chromatography-mass spectrometry]. AB - OBJECTIVE: To investigate lipid components of high-yielded Pinellia ternata rhizomes growing in the west of Hubei province. METHODS: To determine the lipid chemical components in Pinellia ternata rhizomes with GC-MS method and NIST atlas. RESULTS: Ten components have been found: palmitic acid (I), 9,12 octadecadienoic acid (II), pyrrolidine,1-(1-oxo-7,10-hexadecadienyl) (III), alpha monpalmitin (IV), 1,3,12-nonadecatriene (V), campesterol (VI), stigmasta-5,22 dien-3-ol (VII), beta-sitosterol (VIII), stigmasta-5,24-dien-3-ol (IX), cycloartenol (X). CONCLUSION: The relative contents of five kinds of phytosterol: campesterol 28.96%, stigmasta-5,22-dien-3-ol 9.24%, beta-sitosterol 50.77%, stigmasta-5,24-dien-3-ol 4.74%, cycloartenol 6.25%. Component II, III, V, VI, IX are the first time reported in Pinellia ternata. PMID- 17918436 TI - [Study on the terpenoids of chemical constituents of Buddleja purdomii (II)]. AB - OBJECTIVE: To study the chemical constituents of Buddleja purdomii W. W. Smith. METHODS: The constituents were isolated and purified by various chromatographic methods and structurally identified by spectral analysis. RESULTS: 7 compounds were obtained as luteolin (I), luteolin-7-O-beta-D-glucoside (II), trans-caffeic acid (III), cis-caffeic acid (IV), beta-stiosterol (V), stigmasterol (VI), nonacosane (VII). CONCLUSION: All these compounds are obtained from this plant for the first time. PMID- 17918437 TI - [Studies on the chemical constituents from the branches and leaves of Cupressus duolouxiana]. AB - OBJECTIVE: To investigate the chemical constituents from the branches and leaves of Cupressus duolouxiana native to China in order to obtain a more comprehensive understanding of its effective components. METHODS: Compounds were isolated and purified by silica gel column chromatography. They were identified and structurally elucidated by spectral analysis. RESULTS: Six compounds were isolated from the ethanolic extracts of Cupressus duolouxiana, which were identified as (7S,8S)-3-methoxy-3',7-epoxy-8,4'-oxy-neoligna-4,9,9'-triol (I), kaempferol (II), quercetin (III), juglanin (IV), quercetin-3'-methoxy-3-O-alpha-L arabinofuranoside (V) and avicularin (VI). CONCLUSION: All compounds are obtained from this plant for the first time. PMID- 17918438 TI - [Experimental study of anti-atherosclerosis potency by lycium seed oil and its possible mechanism]. AB - OBJECTIVE: To study the effect of anti-atherosclerosis of Lycium Seed Oil (Lso) and its possible mechanism. METHODS: The rabbit atherosclerosis model was established by high fat diet, and the TC, TG, LDL-C, HDL-C levels in plasma were examined dynamically. The SOD, GSH-PX, T-AOC activities and the MDA levels in serum were monitored after 8 week's high fat diet. Aorta samples were observed for atherosclerotic extent, and NF-kappaB, TNF-alpha were assessed by immuno histochemical method. The lovastatin was set up as a positive control. RESULTS: contents of HDL-C obviously increased in Plasma of low and high dosage groups and TC, TG, LDL-C levels significantly decreased compared with control group. The SOD, GSH-PX, T-AOC activities up-regulated while the NF-kappaB, MDA and NF-alpha levels decreased in Lycium Seed Oil groups compared with control group. Aortic atherosclerotic extent and area in low dosage and high dosage LSO groups were absolutely smaller than that in high fat diet group. The anti-atherosclerosis potency of Lycium Seed Oil was similar with that of lovastatin. CONCLUSION: Lycium Seed Oil has potent anti-atherosclerosis effects and its anti atherosclerosis potency was similar with The lovastatin. The possible mechanism involve the decreasing of plasma lipids, anti-peroxidation, inhibiting the activation of NF-kappaB and down-regulating the inflammation cytokines of TNF alpha. PMID- 17918439 TI - [Inhibitory effect of Isorhamnetin and Hesperidin on LDL oxidation induced by Cu2+]. AB - OBJECTIVE: To evaluate the inhibitory effect of Isorhamnetin and Hesperidin on the LDL oxidation induced by Cu2+. METHODS: The serum LDL was isolated by the one step density gradient ultracentrifugation. The LDL oxidation was induced by Cu2+ in vitro for different time periods. Isorhamnetin and Hesperidin at 5 micromol/L were added respectively, as the experimental groups, 3 hours before oxidation. The oxidation of LDL in experimental and control groups was identified by measuring A234 , REM, TBARS and protein carbonyls content. RESULTS: The values of A234, REM, TBARS and protein carbonyls formation increased gradually during LDL oxidation induced by Cu2+ in vitro. During LDL oxidation induced by Cu2+ in vitro and incubation with each of Isorhamnetin and Hesperidin, the kinetic changes of A234 , REM, TBARS and protein carbonyls formation showed lag phases of 2-4 h and 2 h respectively, and the corresponding values for each of the agents treated group were reduced by 23.5%-40.4%, 20.5%-37.7%, 18.6-30.3% and 20.1%-52.4% (P < 0.001); and 11.1%-21.2%, 9.2%-28.3%, 13.7%-21.3% and 5.0%-43.8% respectively (P < 0.001, P < 0.01, P < 0.001 and P < 0.05). CONCLUSION: It suggests that Isorhamnetin and Hesperidin can substantially inhibit LDL oxidation, and Isorhamnetin has antioxidation ability stronger than Hesperidin. PMID- 17918440 TI - [Inhibition of Sorbaria sorbifolia on proliferarion of hepatoma HepG-2 cell line]. AB - OBJECTIVE: To investigate the inhibition of Sorbaria sorbifolia on proliferation hepatoma HepG-2 cells, and to explore the antineoplastic mechanism of Sorbaria sorbifolia. METHODS: MTT assay was used to examine the effect of Sorbaria sorbifolia on the proliferation of HepG-2 cell and flow cytometry was used for the cell cycle distribution. Apoptosis of HepG-2 cell was investigated further by means of inverted microscope, HE staining, transmission electron microscopy and expression of apoptosis-associated gene bcl-2 and p53 were determined by immunohistochemical method. RESULTS: Sorbaria sorbifolia inhibited the proliferation of HepG-2 cell and the effects were in a time-and-concentration dependent manner. Sorbaria sorbifolia could also induce apoptosis, the G2/M phase was arrested and the ratio of apoptosis was significantly difference with that of control group, respectively. The expressions of apoptosis-associated gene bcl-2 and p53 had changed significantly, respectively. CONCLUSION: Sorbaria sorbifolia inhibits proliferation of HepG-2 cells via induction of apoptosis and cell cycle arrest. PMID- 17918441 TI - [Assessment of Pueraria lobata isoflavone with self-microemulsifying drug delivery systems in vitro and in vivo]. AB - OBJECTIVE: To evaluate the self-microemulsifying ability and dissolution behavior of pueraria lobata isoflavone in vitro and the pharmacokinetic behavior in rats. METHODS: The self-microemulsifying rate was evaluated by the self microemulsifying time and the self-microemulsifying efficiency was evaluated by the particle size of resultant microemulsions. The plasma concentrations were evaluated by HPLC and dissolution and pharmacokinetic behavior of self microemulsifying drug delivery systems were evaluated by comparison with commercial tablets. RESULTS: The system was self-microemulsified in 2 min and the particle size was less than 50 nm. The dis- solution of SMESC in distilled water was more than 90% at 10 min, while those of the commercial tablet were less than 50% at 120 min. 82% increase in the relative bioavailability was observed for the self microemulsifying drug delivery systems compared with Yufengningxin tablets. Tmax was smaller in the self-microemulsifying drug delivery systems compared with Yufengningxin tablets. CONCLUSION: The self-microemulsifying drug delivery systems can increase drug dissolution in vitro and absorption in vivo significantly. PMID- 17918442 TI - [Effect of cortex moutan on PGI2, TXA2, ET and NO in diabetic rats]. AB - OBJECTIVE: To study the effect of Paeonol on PGI2, TXA2, ET and NO in diabetic rats. METHODS: Streptozocin was used on rats to make diabetic animal models. Different dosage of Paeonol were used on diabetic animal models. PGI2, TXA2, ET and NO were tested after 30 days therapy. RESULTS: Compared with those in the control group, the level of PGI2 increased and the contents of TXA2 and ET decreased in treatment group. Among them, the high dosage group was more obvious (P < 0.01). But the level of No Had no change. CONCLUSION: Paeonol can decrease the ET and TXA2 in diabetic rats, and increase PGI2 in diabetic rats. PMID- 17918443 TI - [Protective effect of Sankang Capsule on immunity-deficiency mice induced by cyclophosphamide]. AB - OBJECTIVE: To study the immunomodulatory effect of Sankang Capsule( SKC) on the immunity-deficiency mice. METHODS: The immunity-deficiency model was induced by intraperitoneal injection (ip) of cyclophosphamide (CTX) at the dose of 100 mg/ kg in mice which were randomly divided into normal control group, immunity deficiency model group, SKC treated group (250 mg/kg, 500 mg/kg) and positive control group (500 mg/kg). The number of WBC in peripheral blood, the weight of immune organs, the phagocytosis activity of macrophage, the content of serum hemolysin and the proliferation of T lymphocyte in the spleen were measured. RESULTS: The number of WBC, the weight of immune organs, the phagocytosis activity of macrophage, the content of serum hemolysin and the proliferation of T lymphocyte in the model group were all decreased compared with those of normal control group. After the administration of SKC, the index mentioned above was increased compared with those of model group. CONCLUSION: SKC has a protective effect on the immunity-deficiency mice, which may be related to the increasing of cellular immunity, humoral immunity and nonspecific immunity. PMID- 17918444 TI - [Study on the factors of Suanzaoren decoction action by correspondence analysis combined with uniform design]. AB - OBJECTIVE: To investigate the effect of administration dose, administration duration and the time from finishing the last administration to beginning the wild plain method on Suanzaorren decoction action. METHODS: The number of spontaneous activities was determined to study the influence of the factors Suanzaoren decoction action by correspondence analysis combined with uniform design. RESULTS: According to the correlation degree of each factors, there was positive correlation between administration duration and the number of spontaneous activities, whereas there was negative correlation between the two other factors and the number of spontaneous activities. CONCLUSION: It is ecnoomical and efficient to use correspondence analysis combined with uniform design to study the influence of the three factors on Suanzaoren decoction action. The result may provide the basis for pharmacology and clinic. PMID- 17918445 TI - [Microwave-assisted extraction of Hesperidin from pericarpium citri reticulate]. AB - Microwave-assisted extraction (MAE) method was studied for the extraction of Hesperidin from the pericarpium citri reticulate. Several factors, such as the concentration of extraction solvent, MAE time, MAE temperature, microwave power and liquid/ solid ratios were investigated, and the optimal conditions were obtained. The extraction percentage of hesperidiin was obtained by 2.40%, under 14 minutes extraction in 70% methanol water solvent at 65 degrees C , and the liquid/solid ratios of 25:1 (ml/g). Furthermore, compared with the other extraction methods, the MAE was more timesaving and effective. The result indicated that the MAE method is fast, efficient and energy saving, which is useful for active compounds extraction from orange peel. PMID- 17918446 TI - [Study on the optimum extracting conditions of total alkaloids and aconitine in Aconitum szechenyianum Gay]. AB - OBJECTIVE: To optimize the conditions for extractions of total alkaloids and aconitine from Aconitum szechenyianum Gay. METHODS: Using L16 (4(5)) orthogonal test and univariant methods, the exrtaction conditions of total alkaloids aconitine in Aconitum szechenyianum Gay. were optimized under the guidance of the content determination of total alkaloids with bromophemol blue colorimetry and aconitine with HPLC method, respectively. RESULTS: The obtained optimum condition of total alkaloids was that the material was refluxed in 6 times (m/v) acidic alcohol solution( alcohol: pH 3.0 HAc = 85:15, v/v) for 1 h with 3 times. The condition of aconitine was that the material was refluxed in 4 times (m/v) acidic alcohol solution (alcohol: pH 3. p HAc = 15:85, v/v) for 0.5 h with 3 times. Contents of total alkaloids and aconitine in Aconitum szechenyianum Gay. were 0.980% and 0.109%, respectively. CONCLUSION: Through the test and verify, the optimum extraction methods of total alkaloids and aconitine are rational. PMID- 17918448 TI - [Optimization of extraction parameters and de-protein process for Gastrodia elata. Polysaccharides]. AB - OBJECTIVE: Grope for Optimization of Extraction Parameters and De-protein Process for Gastrodia elata Bl. Polysaccharides (GPS). Provide the theoretical basis for the industrialization production of GPS. METHODS: The effects of extracting temperature, ration between Gastrodia elata Bl. and water, heating time on extraction rate of GPS were studied with orthogonal experimental design. Compared the application of the two de-protein process measures (Sevag and Enzyme-Sevag) in purification technics for Gastrodia elata Bl. RESULTS: The optimum parameters were: extraction 3h at 120 degrees C with 1:40 ratio of Gastrodia elata Bl. to water. The de-protein efficiency of Enzyme-Sevag is better than Sevag. PMID- 17918447 TI - [Study on extraction and purification of active parts from Da Chuan Xiong Fang for treatment of migraine]. AB - OBJECTIVE: To study on the process of extraction and purification of active parts from Da Chuan Xiong Fang for treatment of migraine. METHODS: the process of extraction and purification of active parts with ferulic acid and gastrodin were optimized respectively by direct comparison and orthogonal design. RESULTS: Paste rate of active parts with ferulic acid was. 1.67% and the content of ferulic acid was 5.82%. Paste rate of active parts with gastrodin was 5.38% and the content of gastrodin was 8.45%. CONCLUSION: Active parts from Da Chuan Xiong Fang for treatment of migraine are extracted and purified effectively. PMID- 17918449 TI - [Studies on the extraction process of total saponins from Paris polyphylla Smith]. AB - OBJECTIVE: To optimize the extraction process of total saopnins from Paris polyphylla Smith. METHODS: The single factor test and orthogonal experiment were used to determine the optimum extraction process. RESULTS: The optimum extraction process was obtained as follows: the plant materials were extracted with 70% ethanol twice, respectively with 10BV for 2 hours and then with 8BV the solvents for 1.5 hours. The yield of total saponins could be up to 4.24% and the total extraction rate of Paris polyphylla I and Paris polyphylla II was 93.28%. CONCLUSION: The optimum process obtained is steady, reasonable and feasible. PMID- 17918450 TI - [Determination of tanshinone II A and deoxyschizandrin in danshenwuweizi tablet by RP-HPLC]. AB - OBJECTIVE: to set up a determination of tanshinone II A and deoxyschizaadrin in danshenwuweizi tablet by RP-HPLC. METHODS: two components were separated through Kromasil C18 (250 mm x 4.6 mm, 5 microm) column with methanol-water (85:15) by volume as a mobile phase. The flow rate was 1 ml x min(-1) and the detection wavelength were 270 nm (tanshinone II A) and 254 nm (deoxyschizandrin). RESULTS: The linear range of tanshinone II A and deoxyschizandrin were 0.3232 microg-1.616 microg and 0.525 microg-2.625 microg, respectively. The average recovery was 99.83% with RSD 0.36% for tanshinone II A and 100.2% with RSD 0.72% for deoxyschizandrin respectively. CONCLUSION: The method is simple, accurate and rapid with good reproducibility. It can be used for the quality control of danshenwuweizi tablet. PMID- 17918452 TI - [What kind of round lesion is this? Young man with enlarged neck. Cervical cyst]. PMID- 17918451 TI - [The clinical study on decompensatory cirrhotic patients treated by Bie Jia Jian]. AB - OBJECTIVE: To observe the clinical curative effect on decompensatory cirrhotic patients treated by Bie Jia Jian. METHODS: 98 decompensatory cirrhotic patients were randomly divided into two groups: 49 patients in treatment group and 49 in control group. Both groups were treated with the same western medicine of protecting and supporting liver. Except that, treatment group were treated by Bie Jia Jian. RESULTS: The Contents of AST, ALT, total bilirubin (TB), direct bilirubin (DB), hyaluronic acid (HA), Laminin (LN) , procollagen III (pc III), and type IV collagen (IV.C) in both groups decreased after treatment, and prothrombin time activity (PTA) increased. Among them, the decrease of TB, DB, HA, LN, PC-III and IV-C, and the increase of PTA in treatment group were more obvious than those in control group (P < 0. 05). CONCLUSION: Bie Jia Jian is effective in treating decompesatory cirrhotic patients. PMID- 17918453 TI - [New concept for better compliance. Preventing errors in asthma therapy]. PMID- 17918454 TI - [Socially vulnerable persons are especially at risk. Tuberculosis in Germany--a witness of poverty]. PMID- 17918456 TI - [Coxarthrosis: preserve the patient's joint for long as possible]. PMID- 17918455 TI - [Discussion of the safety of lipid therapy. Low LDL--high cancer risk? (interview by Dr. Beate Schumacher)]. PMID- 17918457 TI - [Pathogenesis of hip osteoarthritis]. AB - Coxarthrosis is the most frequently occurring hip disease, is the most common type of osteoarthritis in general and is the second most frequent cause of chronic invalidity. Causes are inherited hip diseases (e.g. dysplasia), growth diseases (e. g. Perthes' disease, slipped capital femoral epiphysis), injuries (fractured femur, fractures of the femoral head or acetabulum, necrosis of the femoral head), and inflammatory, rheumatic or metabolic diseases. Hip deformities such as hip dysplasia or protrusio acetabuli lead to increased mechanical deterioration as a result of the pathological anatomy and hence to a chronically progressing degeneration (coxarthrosis). Primary coxarthrosis usually begins between the 50th and 60th year of life; the secondary disease can develop even earlier depending on cause. PMID- 17918458 TI - [Manifestations and diagnosis of hip osteoarthritis]. AB - The principal symptom of coxarthrosis is mostly pain in the groin. The patient often has a limping gait. The patient notices an increasing restriction in the mobility of the hips and the ability to walk. It must be borne in mind that knee pain can also be the only primary symptom of hip disease. Standard radiographs should be always taken from two different views. Magnetic resonance imaging and computed tomography are still reserved for special problems in the diagnosis of coxarthrosis. PMID- 17918459 TI - [Conservative and surgical therapy of hip osteoarthritis]. AB - Conservative measures generally have priority over surgical procedures. Firstly, an analgesic and antiphlogistic therapy during the acute pain phase is very effective. Secondly, retaining or improving mobility is important. The indication for surgery for hip arthritis exists when the clinical and subjective symptoms in conjunction with the radiological finding can no longer be adequately controlled by conservative measures. Surgical interventions are basically either joint replacing or joint-saving. The goal of all corrective osteotomies is to improve the hip joint congruency and thus delay the replacement of the hip with an artificial joint. Established prosthetic models show long-term results of 15-20 years with a survival rate of more than 90%. PMID- 17918460 TI - [Black spots before the eyes--mouches volantes?]. PMID- 17918461 TI - [Cardiometabolic high risk patients with abdominal obesity. Frequency and risk factor profile in primary care]. PMID- 17918463 TI - [Satellite practices and medical care centers as new "providers". Can contract physicians protect themselves from this competition?]. PMID- 17918462 TI - [Diabetes mellitus and heart failure]. AB - Chronic heart failure (CHF) in patients with diabetes mellitus (DM) is a condition that is frequent and has a poor prognosis. Diabetes mellitus is an independent risk factor for CHF and vice versa. CHF is found in 10-15% of the patients with DM compared to 3% in individuals without DM. Apart from CHD and hypertension, hyperglycaemia and insulin resistance are directly linked to the development of diastolic dysfunction and to CHF. According to the stepwise diagnostic procedure recommended by the ESC in its guidelines from 2005, if heart failure is suspected, the disease should first be diagnosed by ECG, X-ray, or testing for natriuretic peptide and followed by echocardiography when test results are abnormal. Treatment of CHF in patients with diabetes mellitus is the same as that for nondiabetic patients and includes the use of ACEIs, ARBSs (as an alternative to or in combination with ACEIs), BBs, diuretics (in particular loop diuretics), aldosterone inhibitors and digitalis. Most importantly, meticulous glucose control is a must in patients with diabetes mellitus and CHF to improve prognosis. Contraindications for antidiabetic drugs such as glitazones for CHF NYHA classes I-IV and metformin for NYHA classes III-IV need to be considered in patients with CHF and diabetes mellitus. PMID- 17918464 TI - Proteomics in pediatric research and practice. PMID- 17918465 TI - Advances in pediatric pharmacology, therapeutics, and toxicology. PMID- 17918466 TI - Advances in idiopathic scoliosis in children and adolescents. AB - The understanding of idiopathic scoliosis continues to evolve. At some time in the foreseeable future the term will be oxymoronic. It may eventuate, based on a series of laboratory and genetic tests, that there will be three groups: One in which the curve will never be of significance and regular follow-up is unnecessary. A second group with predictable response to a brace. A third group in which spine fusion is an inevitability. This latter group should be allowed to enjoy childhood and early adolescence unfettered by treatment until such time as surgery is necessary. Until then, efforts must be directed toward early detection, comprehensive evaluation, and best efforts at preventing progression of the curve. To this end, a referral to an orthopedic surgeon for suspected scoliosis never is disparaged. PMID- 17918467 TI - Evaluation of the quality of care in congenital heart surgery: contribution of the Aristotle complexity score. PMID- 17918468 TI - Advances in the diagnosis and treatment of osteoporosis. AB - Although severely low bone density is relatively rare in the pediatric population, it can be a significant problem in many patients with chronic illness. As peak bone formation occurs during adolescence, it is crucial that pediatricians and other care providers for this patient population recognize the significance of attainment of adequate bone. Dietary intake of vitamin D and calcium should be optimized, and correction of underlying causes of poor bone density should occur whenever possible. Assessment of bone density is difficult, as each technology available has problems, and none of the technologies are well associated with fracture risk in pediatric patients. Once diagnosis of severely low bone density is established, treatment options are limited and poorly studied. The benefits of bisphosphonate therapy appear to outweigh the risks in patients with low bone density and frequent fragility fractures, and it appears that most improvement with bisphosphonates occurs within the first 2 to 4 years. Evidence, however, is emerging that once off therapy, bone turnover remains decreased for at least several years. During that time, improvements in bone density are decreased. Many questions remain regarding duration of therapy with bisphosphonate therapy and the long-term effects on the children who receive this medication. Anabolic therapies may become important in the future, but there is currently extremely limited information regarding their use in pediatrics. PMID- 17918470 TI - Immunization update. AB - Although the development and licensure of new vaccines over the last 2 years has generated a lot of excitement as well as debate, there is a lot more to come. Not discussed in this article. licensure of another long-awaited vaccine albeit for use in adults was that for herpes zoster. The second HPV and rotavirus vaccines are awaiting approval in the US. Next in line are the vaccines both prophylactic as well as therapeutic against HIV. Topics of debate over the new vaccines include discussions amongst practices as to the affordability and cost of the new vaccines as well as the ethical debate amongst lawmakers and the general public regarding the rights and wrongs of compulsory vaccination against HPV. Another ongoing discussion is regarding the availability of approved vaccines. Shortages have been seen with several of the childhood vaccines including heptavalent pneumococcal conjugate vaccine, tetravalent meningococcal conjugate vaccine, hepatitis A vaccine, as well as the ongoing saga with influenza vaccines. Across the globe while the struggle against polio continues, there is encouraging news regarding the reduction in measles-related deaths, particularly in Africa. The last few years have indeed been landmark years in infectious disease research as the search continues for better and safer vaccines globally. PMID- 17918469 TI - Shedding light on hypovitaminosis D and rickets. AB - The recent resurgence of the ancient disease of vitamin D deficiency rickets and the widespread presence of hypovitaminosis D across the age spectrum pose significant challenges for today's clinicians. Furthermore, new research into previously unsuspected actions of vitamin D in multiple cell systems offer the possibility that vitamin D will play an increasingly important role in our understanding of a wide variety of disease states. PMID- 17918471 TI - The health of homeless children revisited. AB - To the extent that representative data are available for specific health conditions (eg, under-immunization, asthma prevalence), the authors' data suggest that the gap between the health status of homeless children and housed children in minority, low-income families is narrowing. Studies of the health status of homeless children allow a window into the health status of medically underserved children whose needs may not be readily documented because of their lack of access to the health care system. Although prevalence rates of most of the health conditions discussed in this article exceeded national norms, they were generally consistent with rates characteristic of health disparities based on race ethnicity and income. It must be emphasized that in most instances, children were seen for their first pediatric visit within weeks of entering the homeless shelter system. The health conditions identified were often present before the child and family became homeless. The high prevalence of asthma among homeless children should therefore be a matter of concern to health providers and payors, because the authors' data strongly suggest that this is not confined to children in homeless shelters as a special population. Similarly, childhood obesity predates homelessness (or at least the episode of homelessness during which health care was provided) and as such the authors' data may indicate the extent of this problem more generally among medically underserved children in the communities of origin. These conditions seem to be exacerbated by the specific conditions associated with homeless shelter life. Asthma care, assuming it was previously available, is disrupted when housing is lost, and shelter conditions may have multiple asthma triggers. Nutrition often suffers as a result of inadequate access to nutritious food and cooking facilities in shelters, as indicated by the high rate of iron-deficiency anemia among very young children. It is clear that homeless children in shelters require enhanced access to primary and specialist care. Shelter placement necessarily disrupts prior health care relationships (if any), while simultaneously placing additional stress on the child's physical and emotional well being. A medical home model is strongly recommended to allow for continuous, culturally competent care. Developmental and mental health problems are also more prevalent among homeless children. These conditions may jeopardize life successes. The overcrowding associated with homeless shelters and the housing conditions that frequently precede episodes of homelessness are associated with the higher prevalence of otitis media found among young children. This in turn is associated with developmental delay. Also contributing to the developmental risk associated with homelessness is exposure to DV, which is also frequently an antecedent of homelessness. Developmental surveillance for young, homeless children, monitoring of school attendance and academic performance, and assessment of mental status for homeless adolescents are recommended to facilitate early identification of problems and delivery of necessary interventions. For young children, providers of health care to the homeless should be well networked into the Early Intervention and Preschool Special Education programs in their locality. Given the multiplicity of needs for homeless families, which of course includes help finding affordable housing, health care providers serving this population should also develop linkages with community agencies, including those that can help parents develop the skills necessary for economic self-sufficiency and long-term ability to sustain independent housing. PMID- 17918472 TI - Children and megadisasters: lessons learned in the new millennium. AB - Many specific lessons were learned from recent megadisasters in the United States at the expense of children who suffered from a government and a citizenry that was desperately unprepared to respond to and recover from the disaster's short- and long-term effects. During the 9/11 attacks, the nation learned a new sense of vulnerability as the specter of terrorism was delivered repeatedly to our collective consciousness. As this article has emphasized, children experienced significant and widespread psychological effects from this event, and many did not receive adequate treatment. Hurricane Katrina exploited the weaknesses of an already strained child mental health system and vividly demonstrated the liability of poor preparedness and inadequate communication by both families and governments. The impact of Katrina continues to affect many thousands of children over a year later, as the systems that were intended to care for them have largely moved on. Indeed, there was no mention of Hurricane Katrina, the Gulf Coast, or the storm's survivors in the 2007 State of the Union address by the President. After 9/11 and the unprecedented federal spending that occurred to increase our nation's readiness, it is discouraging that the response to Hurricane Katrina fell so short of what had the potential to be the greatest disaster response and recovery story in the history of our nation. It is unlikely that further uncontained expenditures will solve the problems that were exposed in the Gulf Coast. There is not a solution that money can buy. One need only look a few hundred miles south to the Cuban disaster response system to appreciate where some of our shortfalls lie. Cuba has succeeded where the United States has not in part because its citizens are participants in their own preparedness. They engage their children and their families in preparedness planning and they rely upon other members of their community to strengthen their ability to survive as individuals. The American mentality of "dial 911 in an emergency and wait for help" works only as long as there are enough resources to match the need. In a disaster, this approach has proven to be inadequate over and over again. In America, we are well positioned to be leaders in responding to the needs of children affected by disaster. The resources of our government and the resourcefulness of our people should offer much promise for the future. By analyzing our past shortfalls and taking practical steps to mitigate the existing barriers to preparedness, our children, we hope, will fare much better the next time a megadisaster strikes. Box 7 includes suggestions for national priorities for child disaster care. PMID- 17918473 TI - Dental caries: state of the science for the most common chronic disease of childhood. PMID- 17918475 TI - Benefits and risks of breastfeeding. AB - In the absence of significant, unpreventable risks, breastfeeding should be the norm for the nourishment of human infants and should, therefore, be encouraged for populations in all countries. Continued efforts of international and national agencies and healthcare professionals to aid and abet breastfeeding, reduce the risks that occur in some women during breastfeeding, provide the safest substitutes for human milk when that is necessary, and encourage further research into the posed questions should considerably improve the health of many children. PMID- 17918474 TI - Atopic dermatitis. AB - Atopic dermatitis is an especially common and frustrating condition, and the prevalence is increasing. The disease can adversely affect the quality of life of patients and caregivers. Significant advances in our understanding of the pathogenesis have led to improvements in therapy. Patient and caregiver education, avoidance of potential triggering factors, optimal skin care, and pharmacotherapy offer the potential for good control for most patients. PMID- 17918478 TI - Expecting the worst. How disaster management is taking the kidney community by storm. PMID- 17918476 TI - If legislators don't have to live with Medicare, should they be directing its future? PMID- 17918479 TI - American Renal opens new clinic post-Katrina. PMID- 17918480 TI - Surviving change: Can the ESRD provisions in the SCHIP create a level playing field? PMID- 17918481 TI - Finding a rational approach to ESA therapy--for payers and patients. PMID- 17918482 TI - In control with in-center self-care. PMID- 17918483 TI - Choosing in-center self-care hemodialysis. PMID- 17918485 TI - Delivering better results for patient quality of life. PMID- 17918484 TI - Self-cannulation. An interview with Lynda K. Ball. PMID- 17918486 TI - [Our friend...coffee]. PMID- 17918487 TI - [Side effects of proton pump inhibitors (PPIs)]. AB - The proton pump inhibitors (PPIs) are used very widely in the world and generate important health costs. While they bring an undisputed therapeutic benefit, the side effects of the PPIs were recently revalued by population-based case-control studies. There are known frequent minor side effects such as diarrhea or exceptional hypersensitivity reactions. New concerns recently emerged on prolonged use of PPIs: 1) osteoporosis; 2) pneumonia; 3) enteric infection. These new data are summarized here. New set points between the risk/benefit profile of long-term use of PPIs are discussed. PMID- 17918488 TI - [Liver disease in obese patients]. AB - Due to obesity epidemics, nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) are becoming the main causes of chronic liver disease in western countries. Nonalcoholic steatohepatitis is a potentially progressive disease that may cause cirrhosis. We analysed liver histology in 505 patients at the time of gastric by-pass surgery. Steatosis was present in 92% of these patients, mild (< 30% of hepatocytes) in 46%, moderate (30-60% of hepatocytes) in 30%, and severe in 23%. Insulin resistance, diabetes, elevated ferritin and elevated liver tests were independent predictors of NASH in the liver these obese patients. Early identification of these factors might help to select patients at risk of NASH in whom liver biopsy should be considered. PMID- 17918489 TI - [Drug-induced pancreatitis]. AB - Drug-induced pancreatitis represents 2% of acute pancreatitis. The incidence is rising with more than 260 substances that have been incriminated so far. The important steps for the diagnosis are the exclusion of the other causes of acute pancreatitis, the chronology between the introduction of the drug, the appearance of the symptoms and the resolution of the complaints and the elevation of pancreatic enzymes after discontinuation of the treatment as well as the documentation in the literature of similar cases. The degree of the evidence is classified by the strength of the association (definite, probable and possible) and the number of reported cases. The prognosis is in most cases good, but rare cases of death have been reported. PMID- 17918490 TI - [Immunization guidelines in patients with chronic liver diseases]. AB - Patients with chronic liver diseases and specially with cirrhosis have an impaired immune system and are vulnerable to viral (co-) infections. Vaccination against hepatitis A and B virus (HAV, HBV) is recommended for all patients with chronic liver diseases. However, the efficacy of vaccinations in patients with end-stage liver diseases and transplant recipients is disappointing. As soon as a chronic liver disease is diagnosed patients should be immunized with the common vaccines (flu and pneumoccocci). Due to reduced seroconversion rates, antibody levels should be tested after a booster vaccination. PMID- 17918491 TI - [Detection of hereditary hemochromatosis]. AB - Hereditary hemochromatosis is one of the most common genetic disorders. The prognosis of hemochromatosis is normal when phlebotomy therapy is started prior to manifestation of cirrhosis or diabetes. High ferritin is not always a marker of iron overload and ferritin must thus be coupled with transferrin saturation. Only high transferrin saturation entails a genetic research (HFE or type 1). The identification of rare types of hemochromatosis (types 2-4) is only required in particular cases. The evaluation of the iron overload is now based on hepatic MRI determination rather than liver biopsy. PMID- 17918492 TI - [Minimally invasive treatment of obesity with the intragastric balloon: a review]. AB - Obesity is currently one of the most important problems in public health because of its prevalence and potential complications. The National Institute of Health (NIH) recommends to decrease body weight by about 10% because this initial weight loss can significantly decrease the severity of obesity-associated risk factors. The intragastric balloon (IB) was developed as a temporary aid to obtain such a weight loss and to induce a modification of eating habits. Results are encouraging in terms of weight loss, and improvement of co-morbidity. The best results have been obtained in patients with a body mass index (weight/size2) comprised between 30 and 40 kg/m 2, but superobese patients may also be good candidates for IB therapy before bariatric surgery. Overall complications are estimated to develop in about 3% of cases. PMID- 17918493 TI - [Alcohol-related injuries--an emergency department study in the Lausanne University Hospital]. AB - Alcohol-related injuries are responsible for a large share of the global mortality and morbidity burden. Scant information existed, however, for Switzerland. Based on 3653 injured patients and 3519 patients attending the emergency department of the Lausanne University Hospital for other reasons, alcohol attributable fractions with regard to the alcohol consumption in the 6 hours before the injury were estimated. Among men 17% of all injures were alcohol attributable, and 12% among women. Relative risks increased in dose-response relationship with alcohol intake. Leisure time related injuries were most likely to be alcohol attributable. Most of the alcohol-related injuries occurred at already small ethanol quantities ingested. PMID- 17918494 TI - [Multiple myeloma as a cause of rapid acetabular osteolysis]. AB - Acetabular osteolysis is problem affecting long-term survival of total hip prosthesis. CASE REPORT: a 71 year old man was operated in December 2005 with a total uncemented hip arthroplasty. Immediate evolution was favourable but at 4 months postoperatively he came with the complaints of left back pain irradiating in the left thigh. X-rays revealed the presence of a lytic lesion around the acetabulum with secondary dispalcement of the acetabular cup. Further investigations ruled out infection and confirmed the presence of multiple myeloma. The wear and shear particles of polyethylene or metal are the leading cause of reactive osteolysis in total hip arthroplasty, nevertheless multiple myeloma or metastatic lesions should be included in the differentail diagnosis. PMID- 17918496 TI - [Biomedical ethics?]. PMID- 17918497 TI - [Basel firefighters and lung diseases]. PMID- 17918498 TI - [Pathogenic avian influenza virus in swans]. PMID- 17918499 TI - [Nicolas Sarkozy and pedophilia (2)]. PMID- 17918500 TI - [Social support and palliative care, a continuum]. PMID- 17918501 TI - Long term follow-up of carpal tunnel syndrome during pregnancy: a cohort study and review of the literature. AB - INTRODUCTION: Carpal tunnel syndrome (CTS) occurring during pregnancy is considered to have a short and benign course and very few cases required surgery, however there is no information in literature on long term follow-up. The aim of this study was a systematic review of the literature and to report 3-year follow up after delivery in a sample of pregnant women with CTS. PATIENTS AND METHODS: We enrolled 45 consecutive pregnant women with CTS (mean age 32 years). Diagnosis was based on clinical and neurographic findings. Clinical and electrophysiological severity of CTS were scored according to an ordinal scale and the self-administered Boston questionnaires on symptoms (BQ-SYMPT) and functional status (BQ-FUNCT) of the hand during pregnancy and one-year after delivery. Symptoms were evaluated again by BQ over the telephone three years after delivery. RESULTS: At one-year follow-up BQ-SYMPT and BQ-FUNCT scores improved in 40% of women, did not change in 46.7% and 55.6% and worsened in 13.3% and 4.4%, respectively. Clinical severity was stage 0 (i. e. without symptoms) in 26.7% of women, improved in 6.7%, unchanged in 60% and worse in 6.7%. Electrophysiological severity was stage 0 (i.e. no delay in median nerve conduction) in 17.8%, improved in 20%, unchanged in 57.8% and worse in 4.4%. Only one woman underwent surgery (5 months after delivery), three were treated with local steroid injection before delivery and 18 used a splint, 8 of whom continued to do so periodically after one year. At 3-year follow-up 51% were symptom-free and 49% had anomalous ( > 1) BQ scores, but mean BQ scores improved with respect to those at baseline and one-year follow-up. CONCLUSION: A Pubmed search identified 20 papers in which therapy and follow-up could be deduced. Almost all reported a short follow-up with disappearance of symptoms. Our study confirms that pregnancy-related CTS has a benign course: improvement of symptoms was evident at one- and 3-year follow-up, but about half the women still complained of symptoms 3 years after delivery. Only one woman underwent surgery and 11% still sometimes wore a splint at night. Despite improvement of symptoms, distal sensory conduction velocity of the median nerve improved but remained delayed in 84% of women one year after delivery. PMID- 17918502 TI - Influence of trunk muscle co-contraction on spinal curvature during sitting for desk work. AB - Nowadays, a lot of office workers are forced to sit at a desk for many hours while doing their jobs. While sitting, the pelvis rotates backwardly, and lumbar lordosis is flattened. At the same time, the load on the intervertebral discs and spine increases. Sitting in a slumped position is known to increase disc pressure even more, and to aggravate chronic low back pain (CLBP). Therefore, it is very important to teach workers about the correct sitting posture. In addition, it has been recognized that co-contraction of the deep spine-stabilizing muscles enhances lumbar segmental stability and the sacro-iliac joint. However, little is known about the influence of co-contraction of the trunk deep muscles on spinal curvature during sitting for while doing desk work. The purpose of this study was to compare EMG (electromyographic) activity of the trunk muscles during slump sitting with that during co-contraction of the trunk muscles and to investigate how this co-contraction influences spinal curvature. Ten healthy male volunteers (21.7 +/- 2.5 years old) without CLBP participated in the study. Bipolar surface electrodes were attached to the rectus abdominis, the obliquus externus abdominis, the obliquus internus abdominis, the lower back extensor muscles (L3) and the multifidus on the right side. EMG signals were continuously recorded during slump sitting and co-contraction of the trunk muscles, simulating a desk work sitting posture; i.e., slightly inclined forward. They were amplified, band pass filtered, digitized and stored by a data acquisition system. The average muscle activity values over the five-second sample for each sitting posture were normalized to maximal voluntary contractions (%MVC). While the subjects performed both sitting postures, the curvature of the spine was measured using a new skin surface and hand-held device, the "Spinal Mouse". More significant activities of the trunk muscles, with the exception of the rectus abdominis, were observed during co-contraction of the trunk muscles than during slump sitting The co contraction of the trunk muscles resulted in significantly less lumbar curvature and more sacral angle than during slump sitting. The thoracic curvature showed no significant change during either sitting posture. The results of this study indicated that co-contraction of the trunk muscles during sitting while doing desk work could bring about the correct lumbar curvature, and effectively stabilize the lumbopelvic region, and decrease focal stress on passive structures. PMID- 17918503 TI - The influence of the reference electrode in recording F-waves. AB - OBJECTIVE: To demonstrate that many tibial F-waves can be recorded with the tendon electrode and to declare the importance of the "reference" electrode in F wave recording. METHODS: In eight lower extremities of four healthy subjects, tibial and fibular F-waves were recorded with the belly and tendon electrodes separately, using the opposite foot electrode as a reference site. RESULTS: It was found that the mean tibial F-wave persistence proportions were 51.5% with the belly electrode and 86.7% with the tendon electrode. A greater number of tibial F waves were recorded with the tendon electrode than the belly electrode, while few peroneal F-waves were recorded with the tendon electrode. CONCLUSION: Many of the standard tibial F-waves were recorded with the tendon "reference" electrode. SIGNIFICANCE: This phenomenon might be a clue to the difference of the F-wave persistence between the tibial and the peroneal nerves. PMID- 17918504 TI - Electromyographical study of the iliocostalis lumborum and gluteus maximus muscles during locomotion on a treadmill and in a ground. AB - The aim of this work was to study the potential action of the Iliocostalis lumborum and gluteus maximus muscles by electromyography during locomotion in a ground and on a treadmill in 10 male subjects ages 19 to 25 years old. To the locomotion in a ground, the subjects started running in a stationary place for 15 seconds and then ran 10 meters at an average speed of 6.0 mph. To locomotion on a treadmill it was also set 6.0 mph during 60 seconds. The registers presented here refer to the last 5 seconds to treadmill test and during the first 5 seconds to test in a ground. A LYNX- AI 6010 electromyography containing 6 channels was used for this study. It was established a frequency of 1200 Hz to acquire the electromyography's registers, being the low pass filter 600 Hz and the high pass filter 10 Hz. The configuration of the entrance limits of the signals was established at 2000 microV (high limit) and -2000 (low limit)microV for both muscles on a treadmill and in a ground. The results of the muscular potential actions presented in RMS were greater on a treadmill for both muscles, with the predominance of the gluteus maximus muscles when it was compared in a ground. When they were compared between them, the gluteus maximus muscle was more active than the Iliocostalis lumborum muscle. PMID- 17918505 TI - Electromyographyc evaluation of movements of lower limb in double pulley system equipment: comparison between gastrocnemius (caput laterale) and gluteus maximus. AB - It was evaluated movements of lower limb in the double pulley system equipment on ten male volunteers during contraction of gastrocnemius (caput laterale) and gluteus maximus muscles in the following movements: 1) hip extension with extended knee and erect trunk, 2) hip extension with flexed knee and erect trunk, 3) hip extension with flexed knee and erect trunk, 3) hip extension with extended knee and inclined trunk, 5) hip abduction along the midline, 7) hip abduction with extension beyond the midline, 8) adduction with hip flexion beyond the midline, 8) adduction with hip flexion beyond the midline, and 9) adduction with hip extension beyond the midline. Myoelectric signals were taken up by Lec Tec surface electrodes connected to a 6-channel Lynx electromyographic signal amplifier coupled with a computer equipped with a model CAD 10/26 analogue digital conversion board and with a specific software for signal recording and analysis. We observed weak gastrocnemius muscle activity for all movements studied. In the case of gluteus maximus, the most important potentials were observed for movement 2, while for the remaining movements the actions were of reasonable intensity. Compared to gluteus, gastrocnemius was less required for all movements. PMID- 17918506 TI - Tibial and peroneal nerve conduction studies in ankle sprain. AB - BACKGROUND: Lower limb injuries and ankle sprain occur frequently among football players, due to the high incidence of physical contact. As a result, lower limb nerves are subject to injury. This project aims to evaluate the lower limb nerve conduction among students playing football to investigate the probable relationship between ankle sprain in football and nerve conductivity. MATERIALS AND METHODS: Fifty volunteer students aged between 19 and 25 were studied. They fell into three groups; one comprising of 20 healthy football players whose football experience exceeded three years, the second group with 15 football players who had suffered ankle sprain and the last group with 15 healthy non- sports students. Initially, the surface temperature of the foot skin was recorded. Then, the latency and the conduction velocity of deep peroneal and tibial nerves were recorded, statistical data analysis was conducted using statistical tests, i.e., "Independent sampleT", and "Paired T test". RESULTS: In the football player group, student with Hx of ankle sprain, the deep peroneal and tibial nerves distal latencies were significantly longer than the other two groups (P < or = 0.05). In addition, the nerve conduction velocity of the deep peroneal motor and tibial nerves showed a significant decrease in comparison with the other two groups (P < or = 0.05). CONCLUSION: Harming the lower limb nerves, football may increase the nerve latencies and hence decrease the conductivity in lower limbs. Therefore, in electrophysiologic tests of the football players lower limb nerves, especially those with lower limb injuries, one has to consider the fact that the decrease in the clinical neuroconductivity may be preexistent. Thus care should be taken in diagnosing neuropathy, in this group. PMID- 17918507 TI - Newborn hearing screening in Kuwait. AB - OBJECTIVES: The purpose of the study was to detect permanent hearing loss in a universal newborn hearing screening (UNHS) program that included 200 normal ("well baby") newborn children and 15 children at high risk for hearing loss at Ahmadi Hospital in Kuwait. PATIENTS AND METHODS: Distortion product oto-acoustic emissions (DPOAEs) were conducted in the ward on newborns at the age of 2 days. Newborns who did not pass the DPOAE the second time (at the age of 1 month) were evaluated by a brainstem evoked response audiometry (BERA) test within the age of 3 months. If the infant failed the BERA test, the test was repeated at the age of 6 months to confirm any permanent hearing loss. RESULTS: The peak latencies and interpeak intervals of BERA were established for newborns (5-90 days) at our clinic (ms), I: 1.58 + 0.11, III: 4.16 + 0.22, V: 6.70 + 0.24, I-III: 2.57 + 0.19, III-V: 2.59 + 0.24, I-V: 5.12 + 0.26. The incidence of permanent hearing loss was different across nursery levels. In the "well baby" group, 1% had profound sensorineural hearing loss, 1% had severe (70 dBnHL) sensorineural hearing loss, and 98% had normal hearing level. In the "high-risk" group, 26.67% had profound sensorineural hearing loss, 20% had moderate (60 dBnHL) sensorineural hearing loss, and 53.33% had normal hearing level. CONCLUSIONS: The newborn hearing screening program revealed that hearing loss in the "well baby" group is 2%, while in the "high risk" group it is 46.67%. The results warrant the establishment of UNHS programs in Kuwait and other countries in the Middle East to detect permanent hearing loss very early in life and provide appropriate medical treatment. PMID- 17918508 TI - Event related potentials in children of alcoholics. AB - Assessment of ERPs (Event Related Potentials) is a special area of interest in research on vulnerability to alcoholism in human subjects. ERP not only provide information about potential neurofunctional anomalies in healthy individuals, but also relate those neurofunctional characteristics to the cognitive process involved. The aim of the present study is to evaluate the effects of chronic alcoholism and alcoholism risk on children of alcoholic fathers by using ERP parameters. 24 children of alcoholic fathers (9 boys, 15 girls) with a mean age of 18 +/- 3 (range: 15-25) and 17 control subjects (children of non-alcoholic fathers with out a family history of alcoholism) were included to the study. The age range was from 15 to 25 (mean: 21 +/- 3). N200 potential latency recorded from the parietal electrode position was significantly prolonged (p = 0.032) and amplitudes of P200 potential also recorded from the parietal region was significantly low (p = 0.043) relative to controls. However, the rest of the event-related potential parameters including P300 latency and amplitudes recorded from FZ, CZ, PZ electrode positions did not differ significantly from the children of non-alcoholic fathers. The difference in our results from the previous studies may be due to various factors. Genetic, gender, environmental, educational and social factors may have an overall effect on ERP and we believe these factors may be the cause of the differences seen in our study when compared to the previous ones. We believe the gender differences in our group may have had effected the overall results. Consecutive studies with more subject participation are needed to confirm and settle this issue. PMID- 17918509 TI - [Joicelyn--a girl with tuberculous meningitis]. PMID- 17918510 TI - [Hygienic aspects in handling breast milk]. PMID- 17918511 TI - [Breaking through silence. Children of mentally ill parents]. PMID- 17918512 TI - [Treatment and nursing of the child with burn injuries]. PMID- 17918513 TI - [Vacation pass under free sky. Retrospective view of a successful leisure time opportunity]. PMID- 17918514 TI - [Between tears, rage and gratitude. Parents of premature infants on the intensive care unit. An analysis]. PMID- 17918515 TI - [Pediatric hospice forum: meeting point for experts and active personnel]. PMID- 17918516 TI - [Venous catheter associated septicemias. Stepwise reduction by applying hygiene measures on the intensive care unit]. PMID- 17918517 TI - [A different form of pediatric nursing]. PMID- 17918518 TI - An observation of assisted living environments: space use and behavior. AB - Assisted living facilities have become increasingly popular for older adults needing assistance. They are intended to enable privacy and provide support, but the extent to which they do so, and the degree to which these relate to residents' needs, are unknown. This observational study of 1830 residents in 182 facilities indicates that, during the mid-afternoon, the majority of residents are awake (79%), and one-half (49%) are awake and in public spaces. Residents who are cognitively and functionally impaired are more likely to be in public spaces, but less likely to be engaged. Residents who are awake and alone in private spaces are less likely to be impaired, but more likely to have medical conditions. Thus, residents needing more oversight seem to be positioned to obtain that oversight. PMID- 17918519 TI - The effect of housing on perceptions of quality of life of older adults participating in a medicaid long-term care demonstration project. AB - As the nation struggles with the great increase in the numbers of older adults, many questions arise about how to provide housing and long-term care options that will ensure the quality of life of older adults. This study demonstrates that older adults and their families perceive quality of life more positively once moved from a nursing home to an assisted living facility using Medicaid funds. Results of this exploratory study are promising and suggest that having housing options available across the continuum of care with individualized case management offers older adults the hope for "quality living". PMID- 17918520 TI - Harriet Tubman's last work: the Harriet Tubman Home for Aged and Indigent Negroes. AB - This article focuses on the important contributions the venerable Harriet Tubman made to the field of housing for older persons and other populations at risk. It uses an historical approach to document the importance of early housing and self help initiatives in the African American community. It embraces Harriet Tubman and other early housers for their good works and acknowledges them as contributors to the rich legacy of community social work practice and its sage principles of empowerment and self-help. The article presents a nexus between the current housing status of older Blacks and the double jeopardy status imposed by historical discrimination. PMID- 17918521 TI - Chiari malformation type I. PMID- 17918522 TI - Skull base growth in children with Chiari malformation Type I. AB - OBJECT: The goal of this study was to establish whether children with Chiari malformation Type I (CM-I) have abnormal skull base geometry. METHODS: Distances and angles between skull base landmarks were measured on preoperative magnetic resonance images obtained in 30 children (age range 36-204 months) with symptomatic isolated CM-I; 16 of them (53%) had syringomyelia. Comparisons were made with 42 children of similar age who comprised the control group by using one way analysis of variance. RESULTS: The angle formed by the crista galli (CG), dorsum sellae (DS), and foramen magnum (FM) was larger than normal in individuals with CM-I than in those without (145 degrees in patients with CM-I but no syringomyelia and 151 degrees in those with CM-I and syringomyelia compared with 135 degrees in controls; p = 0.000). The angle formed by the left internal auditory meatus (IAM), FM, and right IAM was also larger than normal in the patients (122 degrees in patients with CM-I but no syringomyelia and 123 degrees in those with CM-I and syringomyelia compared with 110 degrees in controls; p = 0.001). The angle formed by the anterior clinoid process (ACP), CG, and right ACP was smaller than normal (29 degrees in all patients with CM-I compared with 34 degrees in controls; p = 0.000). The distance between the two IAMs was longer than normal (75 mm in patients with CM-I but no syringomyelia and 63 mm in those with CM-I and syringomyelia compared with 58 mm in controls; p = 0.000). The distance between the two ACPs was shorter than normal in the syringomyelia group (31 mm in patients with CM-I but no syringomyelia and 27 mm in those with CM-I and syrinx compared with 32 mm in controls; p = 0.001). Within the group of patients with CM-I, the DS-FM and left ACP-right ACP distances were smaller in the syringomyelia group (p = 0.009 and p = 0.037, respectively). CONCLUSIONS: Children with CM-I have abnormal geometrical measurements of their entire skull base, not only the posterior fossa, irrespective of presence of syringomyelia. This may indicate a mesodermal defect as a possible cause of the malformation. PMID- 17918523 TI - Results of a long-term follow-up after neuroendoscopic biopsy procedure and third ventriculostomy in patients with intracranial germinomas. AB - OBJECT: The authors report the results of long-term follow-ups in 12 patients with intracranial germinomas who underwent neuroendoscopic procedures before chemotherapy and radiotherapy, and discuss the usefulness and safety of these procedures. METHODS: Between January 1996 and December 2005 at Kyushu University Hospital, 12 patients with intracranial germinomas underwent neuroendoscopic biopsy procedures involving a flexible fiberscope. Eight patients simultaneously underwent endoscopic third ventriculostomy (ETV) for existing obstructive hydrocephalus. All patients received chemotherapy and radiotherapy postoperatively, according to the regimen promulgated by the Japanese Pediatric Brain Tumor Study Group. The patients were followed for an average of 78.6 months (range 15-134 months), and a retrospective study was conducted. RESULTS: Germinomas were histologically verified in all patients. No postoperative deaths or permanent morbidity was related to the neuroendoscopic procedures. No other cerebrospinal fluid diversion, such as that achieved with a ventriculoperitoneal shunt, was needed for the management of hydrocephalus. A complete response to postoperative chemotherapy and radiotherapy was achieved in all cases. Only one patient had a recurrent lesion in the spinal cord 6 years after the initial treatment; however, this patient had undergone only the neuroendoscopic biopsy procedure without ETV. CONCLUSIONS: Neuroendoscopic procedures can permit a precise histological diagnosis of intracranial germinomas and are safe and effective in the management of hydrocephalus associated with these tumors. The risk of tumor dissemination due to the neuroendoscopic procedures appears to be minimal when the appropriate chemotherapy and radiotherapy are provided postoperatively. PMID- 17918524 TI - Malignant progression in choroid plexus papillomas. AB - OBJECT: Malignant progression of choroid plexus papillomas has been occasionally reported, but this issue has not yet been systematically addressed. METHODS: Frequency and extent of malignant progression were examined in a retrospective series of 124 primary choroid plexus papillomas using uniform histological criteria. RESULTS: After gross-total resection and a mean follow-up period of 59 months, 12 recurrences necessitating neurosurgical intervention had occurred in the 103 cases of choroid plexus papilloma (World Health Organization [WHO] Grade I) and 21 cases of atypical choroid plexus papilloma (WHO Grade II). The proportion of recurring tumors was higher in cases of atypical choroid plexus papilloma than in cases of choroid plexus papilloma (six [29%] of 21 compared with six [6%] of 103, respectively; p < 0.05). In the majority (10 of 12) of the recurrences, there was a close correspondence between the primary tumor and the recurrence with respect to features identified on routine histological examination as well as Ki 67 (MIB-1) proliferation indices (median value 4% for both primary and recurrent tumors; range 0-15% for primary compared with 0-12% for recurrent tumors). However, two patients experienced a transition from a choroid plexus papilloma (WHO Grade I) and an atypical choroid plexus papilloma (WHO Grade II) to choroid plexus carcinomas (WHO Grade III). CONCLUSIONS: Recurrent tumor growth after gross-total resection is rare in choroid plexus papillomas, but malignant progression to choroid plexus carcinoma does occur in a small percentage of tumors. PMID- 17918525 TI - Multislice spiral computed tomography for pediatric intracranial vascular pathophysiologies. AB - OBJECT: Spiral computed tomography (SCT) and, more recently, multislice SCT (MSCT) angiography have established roles in studying subarachnoid hemorrhage (SAH). Potential advantages in MSCT angiography include rapid acquisition, ready availability, ease of monitoring, high spatial resolution, some temporal resolution, and relative freedom from artifacts. The authors assert that these attributes make MSCT angiography the initial imaging method of choice in the assessment of not just SAH but all intracranial vascular pathophysiologies, particularly in children. METHODS: The installation of a MSCT unit sparked the authors' interest in using MSCT angiography and MSCT venography in cases in which they would have formerly performed magnetic resonance (MR) angiography, MR venography, or catheter angiography as an initial investigational method. They retrospectively evaluated seven cases in which they had used the former imaging techniques to study intracranial vascular pathophysiologies. All scans were obtained on a Siemens Sensation 16-slice scanner, and postprocessing was performed on a Leonardo Workstation. RESULTS: Multislice spiral CT consistently provided useful vascular imaging of a wide variety of intracranial vascular pathophysiologies and an alternative imaging modality in patients considered to be too unstable for more time-consuming investigations. CONCLUSIONS: Multislice spiral CT offers advantages over MR imaging in the assessment of intracranial vascular pathophysiologies and frequently allows complete avoidance or deferral of catheter angiography. PMID- 17918526 TI - Predictive value of cerebrospinal fluid parameters in neonates with intraventricular drainage devices. AB - OBJECT: Infection is a common and potentially devastating complication following placement of ventriculoperitoneal (VP) shunts and cerebrospinal fluid (CSF) reservoirs in neonates. The goal of this study was to determine the normal ranges for cell count parameters in neonates with VP shunts and CSF reservoirs, as well as to determine the predictive value of CSF parameters as markers of infection. METHODS: The authors evaluated neonates from 150 different neonatal intensive care units of the Pediatrix Medical Group who had undergone a lumbar puncture, VP shunt insertion, or CSF reservoir placement between 1997 and 2004. Data were collected from 9704 neonates with a mean birthweight of 2573 g and a mean gestational age of 35 weeks. Of these neonates, 181 had VP shunt insertions or CSF reservoir placements. RESULTS: In neonates with negative CSF cultures, significant differences were found between those with and without VP shunts or CSF reservoirs when comparing red blood cell (RBC) count (620/mm' compared with 155/mm3, p < 0.05), absolute eosinophil count (4/mm3 compared with 2/mm3, p < 0.001), protein levels (179 mg/dl compared with 115 mg/dl, p < 0.001), and glucose levels (27.5 mg/dl compared with 49 mg/dl, p < 0.001). No significant difference was found between white blood cell (WBC) counts in neonates with or without VP shunts who had negative CSF cultures. The sensitivity and specificity of a cutoff value of 20 WBCs/mm3 for diagnosing meningitis in neonates with positive cultures and intraventricular drainage devices were 67% and 62%, respectively. CONCLUSIONS: Although differences exist between CSF parameters found in neonates with or without VP shunts or CSF reservoirs, only the difference in RBC count is large enough to be clinically significant. The authors found that the utility of CSF parameters in neonates with VP shunts or CSF reservoirs was limited due to poor diagnostic sensitivity and specificity. PMID- 17918527 TI - Treatment of cerebrospinal fluid shunt infections in children using systemic and intraventricular antibiotic therapy in combination with externalization of the ventricular catheter: efficacy in 34 consecutively treated infections. AB - OBJECT: There are no randomized studies comparing the efficacy of different antibiotic regimens for the treatment of cerebrospinal fluid (CSF) shunt infections, and in the studies that have been reported, efficacy data are limited. The aim of this study was therefore to report the authors' experience using a specific protocol for the management of shunt infections in children. Standard treatment included a two-stage procedure involving externalization of the ventricular catheter in combination with intraventricular and systemic administration of antibiotic medication followed by shunt replacement. Intraventricular treatment consisted of daily instillations of vancomycin or gentamicin with trough concentrations held at high levels of 7 to 17 mg/L for both antibiotic agents. METHODS: During a 13-year study period, the authors treated 34 consecutive intraventricular shunt infections in 30 children. Infections with coagulase-negative staphylococci predominated, and Gram-negative bacterial infection occurred in five children. Ten of the children were initially treated with intravenous antibiotic therapy for at least 3 days, but this treatment did not sterilize the CSF. After externalization of the ventricular catheter, high-dose intraventricular treatment was given for a median of 8 days (range 3-17 days) before shunt replacement. RESULTS: The CSF was found to be sterile (cultures were negative for bacteria) in one of three, seven of eight, 20 of 20, and six of six cases after 1, 2, 3, and more than 3 days' treatment, respectively. In no case was any subsequent culture positive after a negative result had been obtained. Clinical symptoms resolved in parallel with the sterilization of the CSF. There were no relapses or deaths during the 6-month follow-up period, and there have been none as of April 2007. CONCLUSIONS: Despite the ventricular catheter being left in place and the short duration of therapy, the treatment regimen described by the authors resulted in quick sterilization of the CSF, a low relapse rate, and survival of all patients in this series. PMID- 17918528 TI - Ventriculosubgaleal shunts at Columbus Children's Hospital: Neurosurgical implant placement in the neonatal intensive care unit. AB - OBJECT: The authors review all cases in which ventriculosubgaleal (VSG) shunts were placed at Columbus Children's Hospital for the treatment of posthemorrhagic hydrocephalus in order to assess the surgical procedure, effectiveness of surgery, and complications of cerebrospinal fluid diversion to the subgaleal space. The purpose of the review is to make a comparison between cases in which shunts were placed in the operating room (OR) and those in which they were placed in the neonatal intensive care unit (NICU). Considerations and complications specific to patient transport to the OR or surgical implantation in the NICU are discussed. METHODS: Seventeen infants with posthemorrhagic hydrocephalus were treated with VSG shunt placement over a period of 4 years. A retrospective analysis of these cases was performed to evaluate multiple aspects of the procedure. Specifically, the surgical procedure, duration of shunt function prior to shunt conversion, neuroimaging changes, operative complications, and risk of infection are discussed. The authors also performed a comparative analysis of shunt placement in the NICU and the OR. RESULTS: The length of the procedure was similar in the two locations. No differences in perioperative or intraoperative risks and no increased risk of infection were seen in either location in this pilot study. Interestingly, the mean lifespan of primary implants placed in the NICU (73 days) was longer than that of those placed in the OR (43 days). CONCLUSIONS: Ventriculosubgaleal shunt placement offers a safe and effective temporary means of treating post-hemorrhagic hydrocephalus and can be reliably and safely performed at the bedside. PMID- 17918529 TI - Intraventricular baclofen. PMID- 17918530 TI - Long-term intraventricular baclofen infusion in beagles. AB - OBJECT: This study was designed to evaluate the clinical and histopathological effects of long-term (3-month) intraventricular baclofen (IVB) infusion in beagle dogs. METHODS: Catheters were inserted stereotactically into the lateral ventricles of nine dogs and were connected to implanted Synchromed II pumps. Saline solution (control animals) and IVB (experimental animals) were infused continuously. The IVB dosages ranged from 100 to 1000 microg/day. RESULTS: An IVB infusion of 135 microg/day or less throughout a 3-month period was associated with no adverse side effects in any animal. Infusion of 150 microg/day produced overt seizures in one dog and produced adverse side effects in another; a reduction in the dosage given to these animals to 135 microg/day was tolerated with no adverse effects. Infusion of IVB at a dosage of 250, 500, or 1000 microg/day caused lethal toxicity within the first 4 days of infusion. Histopathological specimens obtained at necropsy revealed no ventricular or subependymal changes in animals receiving an IVB dosage of 135 microg/day or less. CONCLUSIONS: Intraventricular baclofen infusion in beagles has dose-related toxicity; however, no clinical or neurological toxicity was evidenced at clinically tolerated dosages (< or =135 microg/day) throughout 3 months of infusion. PMID- 17918531 TI - Dynamic contrast-enhanced magnetic resonance angiography of vascular malformations in pediatric patients. Case report. AB - The authors report the first clinical use of 3-tesla dynamic contrast-enhanced magnetic resonance (MR) angiography for the diagnosis of a vascular malformation in a pediatric patient. The supply and drainage of an arteriovenous malformation were accurately demonstrated on MR angiography, which was performed without sedating the patient. This lesion was confirmed on catheter angiography, and definitive treatment via embolization was undertaken in a single session. The patient's therapeutic response will be followed with surveillance dynamic MR imaging. PMID- 17918532 TI - Coil embolization of ruptured middle cerebral artery aneurysms in the first 2 months of life. Report of two cases. AB - Intracranial artery aneurysms are very rare in infants. There have been no previous reports of coil embolization to branches of the middle cerebral artery (MCA) in infants. The authors describe successful embolization of ruptured aneurysms of the right frontoopercular MCA branch in a 31-day-old infant, and of the left sylvian MCA branch in a 54-day-old infant. One of the cases involves a dissection flap. The authors also review cases in which coil embolization was used for intracranial aneurysms in the first 2 months of life. PMID- 17918533 TI - Acute hydrocephalus secondary to obstruction of the foramen of monro and cerebral aqueduct caused by a choroid plexus cyst in the lateral ventricle. Case report. AB - Choroid plexus cysts are common and typically asymptomatic abnormal folds of the epithelial lining of the choroid plexus. Rarely, these cysts may gradually enlarge and cause outflow obstruction of cerebrospinal fluid. The authors present a case of a large choroid plexus cyst causing acute hydrocephalus in a previously healthy 2-year-old boy. The patient presented with markedly declining mental status, vomiting, and bradycardia over the course of several hours. Computed tomography scans demonstrated enlarged lateral and third ventricles with sulcal effacement, but no obvious mass lesions or hemorrhage. There was no antecedent illness or trauma. A right frontal external ventricular drain was placed in the patient, resulting in decompression of only the right lateral ventricle. Magnetic resonance (MR) imaging demonstrated a lobulated cyst arising from the choroid plexus of the left lateral ventricle and herniating through the foramen of Monro into the third ventricle, occluding both the foramen of Monro and the cerebral aqueduct. The patient underwent an endoscopic fenestration of the cyst, and histological results confirmed that it was a choroid plexus cyst. Postoperative MR imaging showed a marked reduction in the cyst size. The cyst was no longer in the third ventricle, the foramen of Monro and the aqueduct were patent, and the ventricles were decompressed. The patient was discharged home with no deficits. To the authors' knowledge, there are no previous reports of a choroid plexus cyst causing acute hydrocephalus due to herniation into the third ventricle. This case is illustrative because it describes this entity for the first time, and more importantly highlights the need to obtain a diagnosis when a patient presents with acute hydrocephalus without a clear cause. PMID- 17918534 TI - Cutaneous manifestations of split cord malformations. Report of three cases. AB - A split cord malformation (SCM) is a rare congenital disorder involving a division of the spinal cord that results in two distinct hemicords divided by bone, cartilage, or a fibrous septum. Cutaneous manifestations of SCMs are common, including hypertrichosis, dimples, hemangiomas, nevi, lipomas, and sinus tracts. In this report, the authors describe cutaneous manifestations of SCMs in three patients, including a rare report of near fully formed digits overlying the SCM. PMID- 17918535 TI - Spontaneous regression of lipomyelomeningocele associated with terminal syringomyelia in a child. Case report. AB - The authors present the case of a 3-month-old girl who presented with lipomyelomeningocele (LMM) associated with a low-lying conus and a terminal syrinx. At the time of presentation, the girl had a mild neurological, but congenital, deficit: weakness of the right foot and calf as well as absence of the right ankle reflex. Bladder assessment was normal, and it was decided to monitor this child closely, without performing surgery. Results of further magnetic resonance imaging performed 1 year after diagnosis and when the girl was 28 months old, showed a significant regression of LMM and associated syrinx. Interestingly, the child remained neurologically unchanged. Although described in the literature, spontaneous regression of LMM is a very unusual situation, and very few cases have been reported. This case represents the first report of concomitant regression of an associated syrinx. Close follow-up of nonsurgically treated children is mandatory, as the risk of late deterioration remains. PMID- 17918536 TI - Bobble-head doll syndrome associated with Dandy-Walker syndrome. Case report. AB - Bobble-head doll syndrome (BHDS) presents in childhood and is usually associated with lesions of the third ventricle. This disorder is characterized by stereotypical head movements of the type "yes-yes" (up and down) at a frequency of 2 to 3 Hz. Rarely, movements of the type "no-no" (side-to-side) are described. There are a few hypotheses to explain the mechanism responsible for BHDS, but its real pathophysiological characteristics are still unknown. The authors describe the case of a child born with hydrocephalus and Dandy-Walker syndrome. A ventriculoperitoneal shunt was implanted in the child because of progressive head enlargement. One year after shunt placement, she began making frequent horizontal head movements of the type "no-no". There were no other signs or symptoms. Imaging studies demonstrated small ventricles and a posterior fossa cyst with no signs of hypertension. The child's growth, development, and head circumference (within the 5th percentile) remained satisfactory. Three aspects of this case were of interest: the association of BHDS with Dandy-Walker syndrome, the rare occurrence of BHDS of the "no-no" type, and the absence of third ventricle dilation. The authors' findings support the hypothesis that cerebellar malformations themselves can PMID- 17918538 TI - Choroid plexus hyperplasia: surgical treatment and immunohistochemical results. Case report. AB - Diffuse villous hyperplasia of the choroid plexus is a rare but potential source of nonobstructive hydrocephalus. In addition to discussing the authors' staged surgical approach and medical management decisions in a patient with this rare and challenging condition, immunohistochemical studies of the choroid plexus epithelium are presented to examine the pathophysiological factors involved in abnormal cerebrospinal fluid (CSF) production in this disease. The patient, a 15 month-old girl born at 36 weeks' gestation, underwent a bilateral craniotomy with resection of the choroid plexus to treat her villous hyperplasia. Immunohistochemical studies of the resected choroid plexus were conducted for the purpose of examining the carbonic anhydrase II (CAII) enzyme and the aquaporin 1 (AQP1) membrane protein. Results were compared with immunohistochemical studies conducted in a small series of autopsy specimens of normal human choroid plexuses. There was no change in the immunoreactivity of CAII in the patient with villous hyperplasia compared with normal controls, whereas AQP1 immunoreactivity was significantly weaker in the patient compared with normal controls. Postoperatively, the patient's CSF overproduction resolved and her neurological symptoms improved over time. Shunting techniques and presently available pharmaceutical treatments alone do not provide adequate treatment of high-output CSF conditions. Surgical removal of the affected choroid plexus is a feasible and effective treatment. Results of the immunohistochemical studies reported here support the suggestion that the CAII enzyme is retained in villous hyperplasia of the choroid plexus. However, there appears to be decreased expression and perhaps downregulation of AQP1 in villous hyperplasia compared with normal choroid plexus. Future studies may elucidate the significance of these observations. PMID- 17918537 TI - Neuromyelitis optica with brainstem lesion mistaken for brainstem glioma. Case report. AB - Neuromyelitis optica (NMO) is a severe demyelinating syndrome defined principally by its tendency to affect optic nerves and the spinal cord selectively. Asymptomatic brain lesions have recently become a common finding in NMO, and symptomatic brain lesions do not exclude the diagnosis of this entity. The authors describe the case of a 12-year-old girl suffering from an unusually atypical form of NMO in which a brainstem lesion was mistaken for a brainstem glioma. Brainstem involvement in NMO exhibits variable features on neuroimaging and is confused with brainstem glioma in cases of extensive brainstem involvement in childhood. Careful differential diagnosis and proper treatment are vital for a favorable prognosis. PMID- 17918539 TI - Teratoma within an encephalocele: common etiology or coincidence. Case report. AB - Encephalocele is a cystic congenital malformation in which central nervous system structures herniate through a defect in the cranium. The coincidence of encephalocele and neoplasm is a very rare condition. The authors present the case of a 5-month-old girl who underwent surgery for encephalocele and whose postoperative histopathological assessment revealed evidence of a mature teratoma inside the lesion. The embryogenesis of such a lesion is discussed. PMID- 17918540 TI - Transpalatal endoscopic endonasal resection of a giant epignathus skull base teratoma in a newborn. Case report. AB - Teratomas are neoplasms composed of tissues from all three germ layers with varying degrees of differentiation. They are most commonly found in the sacrococcygeal and gonadal regions and rarely occur in the head and neck region. A teratoma is termed "epignathus" when it arises from the skull base or hard palate and is located in the oral cavity. The authors describe a case of a giant epignathus teratoma originating in the skull base of a neonate, extending bilaterally via two pedicles throughout the hard palate and protruding through the oral cavity. The tumor was completely resected using a transpalatal endoscopic endonasal approach. The excised tumor proved to be an immature teratoma with well-differentiated yolk sac elements. At the 1-year follow-up the patient showed no evidence of tumor recurrence and the child remains neurologically intact. This report demonstrates the use of a transpalatal endonasal corridor in a preterm infant. This approach provided an ample corridor into the ventral skull base without the need for external excisions and/or disruption of osseous elements. PMID- 17918541 TI - Pediatric Cushing disease. PMID- 17918542 TI - Knowledge about suicide and local suicide prevalence: comparison of Estonia and Austria. AB - Knowledge about suicide might be positively correlated with local suicide prevalence. This hypothesis was tested with a sample of 107 medical and social science undergraduates from Estonia, a country with a high suicide rate, using Hubbard and McIntosh's 1992 Revised Facts on Suicide Quiz. Compared (independent groups t tests) with a sample of undergraduates from Austria, which has a markedly lower suicide rate, Estonians presented significantly higher overall knowledge about suicide (d = 0.39), particularly concerning demographic and epidemiological facts about suicide (d = 0.77) but not pertaining to clinical and interpersonal knowledge contents (d = -0.06). Study limitations and suggestions for extension of this preliminary inquiry are discussed. PMID- 17918543 TI - The use of PowerPoint shareware for making Jeopardy!--type games in the teaching of psychology. AB - We describe the use of readily available templates to create a Jeopardy!--type game using PowerPoint slides suitable for psychology courses. The templates can accommodate information from any psychology class and can easily be used to incorporate languages other than English. A major limitation of the templates is that they are only available as shareware on internet sites and the links to such sites can be unreliable. To avoid this problem we present our own template. Differences between the shareware and our own program are noted, as are ways to use the game to stimulate interest in psychology. PMID- 17918544 TI - Motor skills transfer from gymnastics to swimming. AB - 99 adult specialists in combat sports (n = 21), team sports (n = 37), gymnastics (n = 22), and swimming (n = 19) (M age = 20 yr., SD = 2; 64 young men, 35 young women) performed three 25-m swimming tasks whilst "blindfolded" by opaque goggles: front crawl in a straight line, dolphin-kicking on the back, and dolphin kicking on the front. Even though the gymnasts (like the swimmers) were at ease in all 3 situations, the motor skills of the "motor interaction" specialists (team sports, combat sports) put the latter at a disadvantage. The similarities between the gymnasts' and swimmers' behavior (confirmed using factorial correspondence analysis with the TRIDEUX program) are undoubtedly related to the fact that these sportspersons essentially live in the same sensory space in their respective practices: exteroceptive information is subordinated by proprioceptive information. In contrast, the 99 subjects' timed freestyle swimming performances over 75 m depended so much on their physical and anthropometric qualities that the results were independent of the sporting specialties for the novice swimmers. Hence, the mere fact that the 22 gymnasts, who tended to be shorter, with a higher proportion of women, were more at ease in the water did not turn them into great performers. PMID- 17918545 TI - On the nature of the plural self. AB - In a sample of 100 college students, those with a higher score on a measure of having a plural self-concept scored higher on a test of self-monitoring and lower on a test of tolerance for ambiguity, but variance accounted for is small. PMID- 17918546 TI - Club shaft weight in putting accuracy and perception of swing parameters in golf putting. AB - This study assessed how shaft weight influenced golf putting accuracy and subjective perception of swing parameters. Three putters of different shaft weight (100, 420, and 610 gm) were tested by 24 club players. Distance and deviation in direction were measured, and subjective ratings of the putters recorded. Subjects hit the ball further with lighter shafts. The mean distance hit was 100.2, 99.3, and 98.1% of the target distance for the normal, medium, and heavy putter shafts, respectively. Subjectively, the medium heavy putter was rated best on "overall feeling" and it was also rated better than the normal on"feeling of stability in the downswing." The heaviest putter was rated as too heavy by 23 of 24 subjects. There were no significant differences between the putter clubs in distance and directional putting accuracy. The major findings are that the golfers putted 2.1% longer with the 100 gm shaft than with the 610 gm shaft and that the perception of overall feeling of the putter club was not related to performance. PMID- 17918547 TI - Parents' representations of their children: an exploratory study using the Osgood Semantic Differential Scales. AB - The results of an exploratory national study carried out in Italy using Osgood's Semantic Differential Scales (Osgood, Suci, & Tannenbaum, 1957) with parents of 100 normal 6- to 11-yr.-old children in mainstream classrooms are reported. The aim was to devise a simple tool to be used to explore parents' perceptions of their children in this age group. Parents were asked to rate "my child" using some of Osgood's Semantic Differential Scales especially adapted for this study. Participants were 97 fathers and 100 mothers. Fathers were 32 to 57 years of age (M = 43.2, SD = 5.4), mothers were 29 to 49 years of age (M = 39.7, SD =4.4). In factor analysis four factors were identified: Activity, Evaluation, Emotions evoked by the child, Personality/Physical contact. Average factor scores were significantly different. There were no significant differences between fathers and mothers on the four factor scores. Children were more positively evaluated the younger they were. PMID- 17918548 TI - Enabling a young man with minimal motor behavior to manage independently his leisure television engagement. AB - Persons with severe spastic tetraparesis and minimal motor behavior may be confined to a wheelchair or bed and have virtually no chances of constructive engagement with their immediate environment. A possible way to modify this situation may involve the use of technology. The present study (a) assessed specific technology to enable a young adult to manage his leisure television engagement independently and (b) carried out a social validation assessment of the technology-supported performance involving 90 teacher trainees as raters. The intervention period with the new technology included 67 sessions, during which the participant performed independently 392 of the 408 television-management responses, i.e., turning on the television, finding a channel with a preferred program, setting the volume, and turning off the television. He also indicated preference for using the technology as opposed to not using it. The raters provided relatively high (positive) scores for the technology-supported performance compared to the baseline performance. Implications of the findings are discussed. PMID- 17918549 TI - Effects of a computer feedback treatment and behavioral support protocol on drop out from a newly initiated exercise program. AB - Drop out from newly initiated exercise regimens is problematic. Three treatments intended to support new exercise programs (standard exercise counseling, computer feedback, and behavioral support) were tested to estimate their association with drop out over the initial 3 and 6 mo. Data from a total of 1,336 adults (44% men; M(age) = 41.9 yrs., SD = 9.8) initiating exercise programs at 18 community exercise facilities (six in each of the three treatment groups) were aggregated by facility on a measure of drop out and were then contrasted both within and between groups. After a mixed model repeated-measures analysis of variance was significant overall, planned contrasts were conducted using the Tukey-Kramer test. Significantly less drop out was found at Month 3 for both the computer support (27%) and behavioral support (17%) groups, when contrasted with the standard exercise counseling group (38%). Behavioral support had significantly less drop out than computer feedback at Month 3. The behavioral support group showed significantly less drop out at Month 6 (33%) when contrasted with both the standard exercise counseling (58%) and computer feedback (52%) groups, which did not significantly differ from one another. Limitations and the need to evaluate and extend research on interventions for reducing drop out from exercise programs were discussed. PMID- 17918550 TI - Effect of manipulating positive and negative feedback on goal orientations, perceived motivational climate, satisfaction, task choice, perception of ability, and attitude toward physical education lessons. AB - This study examined the effect of different types of feedback on goal orientation, perception of motivational climate, satisfaction, and boredom in physical education (PE) classes, pupils' preferences for challenging versus easy tasks, pupils' attitudes towards PE lessons, and perception of gymnastic ability. 95 subjects in three treatment conditions (positive feedback, negative feedback, and both feedback types) participated in 14 lessons and completed pre- and postintervention measures. Results showed that subjects in the positive feedback group had significantly higher scores on learning-oriented motivational climate and enjoyment than the subjects of the negative feedback group. The negative feedback group reported higher scores on performance-oriented motivational climate than the positive feedback group. Results also showed that subjects in the group receiving both types of feedback had lower scores on learning-oriented motivational climate, higher scores on enjoyment than the negative feedback group, and lower scores on boredom than the positive feedback group. The effects of feedback on physical education students' motivation are discussed. PMID- 17918552 TI - State IQ and suicide rates in the United States. AB - Previous findings pertaining to the direction (positive or negative) of the ecological correlation between intelligence and suicide rates in the USA have been conflicting. Using novel state IQ estimates, derived from the Scholastic Assessment Test, the American College Test, these tests combined, or the National Assessment of Educational Progress state scores, these estimates were not consistently associated to state suicide rates. Whereas ACT-derived state IQ was significantly positively correlated with suicide rates, the correlation with composite ACT-SAT-derived state IQ was significantly negative and with both SAT derived and NAEP-derived state IQ also negative but not significant. Validity checks pointed to possible methodological problems with the state IQ estimates. Currently available estimates of state IQ, therefore, seem not appropriate to resolve the question of the direction of the ecological correlation of intelligence and suicide mortality across the USA. PMID- 17918551 TI - Prediction of goal orientation and perceived competence on intensity and direction of precompetitive anxiety among adolescent handball players. AB - This study explored the main and interactive effects of goal orientations and perceived competence on intensity and direction of the symptoms of precompetitive cognitive and somatic anxiety. 109 handball players from 16 high school teams (M = 16.2 yr., SD = 1.5) participated. All were asked to complete the Task and Ego Orientation in Sport Questionnaire, Perceived Competence Questionnaire, and a modified version of the Competitive State Anxiety Inventory-2. Using separated multiple hierarchical regression analyses, direction of the symptoms of precompetitive somatic anxiety was predicted by perceived competence and interaction of ego orientation x perceived competence. Perceived competence and the interaction of ego orientation x task orientation x perceived competence were significant predictors of direction of the symptoms of precompetitive cognitive anxiety. The results suggest that perceptions of competence and task orientation moderate the relationship between ego orientation and direction of the symptoms of precompetitive anxiety. PMID- 17918553 TI - Eye- and mouth-opening movements replacing head and hand responses in a microswitch program for an adolescent with deteriorating motor condition. AB - This study assessed the possibility of replacing head and hand responses (no longer feasible) with minimal movements within the microswitch-based program of an adolescent with deteriorating motor condition and multiple disabilities. The new movements, i.e., eye- and mouth-opening, were introduced individually and then combined through the simultaneous availability of the related microswitches. Data showed the participant acquired the new movements (responses) successfully and retained them at a 2-mo. postintervention check. Mood improvements, i.e., increases in indices of happiness, also occurred through the program. The conclusion was that a person with deteriorating motor conditions may be able to revitalize a microswitch-based occupational program and retain a constructive engagement if new, feasible responses are identified. PMID- 17918554 TI - A preliminary note: campus subgroup membership and football game day attire. AB - Differences were found in college women's attire at football games by organizational identification, perceived organizational prestige, game day attendance, and fashion opinion leadership. Factors of clothing choice were Fashion Consciousness, Desire for Comfort, Desire for Uniqueness, and Expression of School Spirit. PMID- 17918555 TI - Effects of shape parameters on the attractiveness of a female body. AB - Various researchers have suggested that certain anthropometric ratios can be used to measure female body attractiveness, including the waist to hip ratio, Body Mass Index (BMI), and the body volume divided by the square of the height (Volume Height Index). Based on a wide range of female subjects and virtual images of bodies with different ratios, Volume-Height Index was found to provide the best fit with female body attractiveness, and the effect of Volume-Height Index can be fitted with two half bell-shaped exponential curves with an optimal Volume-Height Index at 14.2 liter/m2. It is suggested that the general trend of the effect of Volume-Height Index may be culturally invariant, but the optimal value of Volume Height Index may vary from culture to culture. In addition to Volume-Height Index, other body parameters or ratios which reflect body proportions and the traits of feminine characteristics had smaller but significant effects on female body attractiveness, and such effects were stronger at optimum Volume-Height Index. PMID- 17918556 TI - Relations of singing talent with voice onset time of trained and untrained female singers. AB - This study examined phonatory-articulatory timing during sung productions by trained and untrained female singers with and without singing talent. 31 untrained female singers were divided into two groups (talented or untalented) based on the perceptual judgments of singing talent by two experienced vocal instructors. In addition to the untrained singers, 24 trained female singers were recorded singing America the Beautiful, and voice onset time was measured for selected words containing /p, b, g, k/. Univariate analyses of variance indicated that phonatory-articulatory timing, as measured with voice onset time, was different among the three groups for /g/, with the untrained-untalented singers displaying longer voice onset time than the trained singers. No other significant differences were observed across the other phonemes. Despite a significant difference observed, relatively small effect sizes and statistical power make it difficult to draw any conclusions regarding the usefulness of voice onset time as an indicator of singing talent. PMID- 17918557 TI - Digit ratios (2D:4D and other) and relative thumb length: a test of developmental stability. AB - Sex and side differences in relative thumb length of children and adolescents have been reported by prior researchers. These findings mirror those reported for the second-to-fourth digit ratio (2D:4D), a likely biomarker for the organizational (permanent) effects of prenatal androgens on the human brain and body. The present study investigated relative thumb length, in particular, its associations with all possible digit ratios (2D:3D, 2D:4D, 2D:5D, 3D:4D, 3D:5D, and 4D:5D), in a sample of young adults (60 men and 64 women). Relative thumb length was less precisely measurable than direct or image-based finger-length measurements, as used in digit ratio research. There were no significant sex or side differences in relative thumb length. Contrary to expectation, thumb size was not positively correlated with any digit ratios. Relative thumb length appears to be developmentally unstable (decreases during childhood and adolescence). Additional findings, such as the magnitude ranking of sex differences in digit ratios and the comparability of direct versus image-based finger-length measurements, are also discussed. PMID- 17918558 TI - Specification of variables predictive of victories in the sport of boxing. AB - Compared to other sports, very little research has been conducted on which variables can predict victory in the sport of boxing. This investigation examined whether boxers' age, weight change from their preceding contest, country of origin, total number of wins, total number of losses, performance in their preceding contest, or the possession of a championship title was predictive of a winning performance in a given bout. A 1-mo. sample of male professional boxing records for all contests held in the USA (N = 400) were collected from the BoxRec online database. Logistic regression analysis indicated that only boxers' age, total number of wins and losses, and the performance in the preceding contest predicted significant variance in outcome. PMID- 17918559 TI - Contrast of frequency of positive and negative feeling state changes associated with standard and reduced cardiovascular exercise. AB - A field investigation was conducted to assess whether cardiovascular exercise assigned at a level to maximize improvements in fitness, i.e., a standard exercise amount or 30 min. at approximately 80% maximal heart rate, differed significantly in frequency of positive exercise-induced feeling changes when contrasted with exercise that was reduced to 20 min. at approximately 65% maximal heart rate. A significantly more frequent occurrence of positive changes in feelings just after exercise (chi1(2), = 4.95, p < .05, phi(c) = .26) was noted for adults initiating programs at the reduced (n = 36; 86% positive change) vs the standard (n = 36; 64% positive change) amount of exercise. After replication, implications for adjusting exercise to increase adherence were discussed. PMID- 17918560 TI - Using developmental principles to assist preschoolers in developing numeracy and literacy. AB - In a yoked control design, 4-yr.-olds (N = 39) in a Head Start program played numerous structured games involving either the oddity principle or letter identification and letter sounds. The children's mean age was 53.2 mo.; SD = 4.1 mo. Three were Middle Eastern, 14 were Latino, 7 were East African, and 15 were African American. Children showed better mastery of oddity after playing games directed at this concept, and numeracy scores on the Woodcock-Johnson III were better for children who had played this type of game. Woodcock-Johnson III Letter Word scores for children who had played the oddity and seriation or letter games were equivalent. These results are consistent with other research indicating that the understanding of oddity relations may be a key transitional thinking which supports quantitative and verbal development at the preschool-kindergarten interface. The standardized test scores indicate that guided play directed at this aspect of cognitive growth or more narrowly directed at early literacy can produce equivalent knowledge of letters. PMID- 17918561 TI - Contextual interference effects with two tasks. AB - The present study was intended to test whether the standard contextual interference effect would be observed when only two patterns were practiced in blocked or random schedules of multisegment movement tasks rather than three patterns which is the norm. In line with the basic effect, Blocked learners' performance was closer to ideal during acquisition, but Random learners had smaller errors at 24-hr. retention. In addition, random learners had better recall of the details of the patterns they had practiced. Learners' predictions of retention performance did not match the group differences actually observed. PMID- 17918562 TI - Assessing the effect of cognitive styles with different learning modes on learning outcome. AB - In this study, similarities and differences in learning outcome associated with individual differences in cognitive styles are examined using the traditional (face-to-face) and web-based learning modes. 140 undergraduate students were categorized as having analytic or holistic cognitive styles by their scores on the Style of Learning and Thinking questionnaire. Four different conditions were studies; students with analytic cognitive style in a traditional learning mode, analytic cognitive style in a web-based learning mode, holistic cognitive style in a traditional learning mode, and holistic cognitive style in a web-based learning mode. Analysis of the data show that analytic style in traditional mode lead to significantly higher performance and perceived satisfaction than in other conditions. Satisfaction did not differ significantly between students with analytic style in web-based learning and those with holistic style in traditional learning. This suggest that integrating different learning modes into the learning environment may be insufficient to improve learners' satisfaction. PMID- 17918563 TI - Regional intelligence and suicide rate: new data for Australia and a synthesis of research. AB - Previous research has shown for the most part positive correlations between intelligence and suicide prevalence on the national level. However, this study found proxies for regional intelligence in Australia (international average domain scores from the PISA 2000 study) to be significantly negatively correlated with the total, male, and female suicide rates of the different administrative divisions of Australia, and this finding was independent of regional wealth. A research synthesis of the current results and those from similar studies of other countries (positive correlations for Austria, Belarus, The British Isles, Denmark, and The Netherlands; inconclusive findings for France, Germany, and the USA) was conducted. This synthesis of research findings showed that positive ecological correlations of intelligence with suicide rate were more likely observed for nations with higher suicide rates and poorer general living conditions, whereas there was no relation with national IQ. PMID- 17918564 TI - Scores for schizotypy and five-factor model of a sample of distant healers: a preliminary study. AB - A preliminary study was conducted to evaluate the personality scores of 18 distant healers on the NEO-Five Factor Inventory and the Schizotypal Personality Questionnaire-Brief. These distant healers differed significantly in their schizotypic tendencies, both in comparison to normed scores and to a control group of 19 masseurs. Further differences were found on scales of Conscientiousness and Openness to experience. PMID- 17918565 TI - On-task distractors associated with totball baseball games: a preliminary study. AB - Two totball baseball leagues in southern Louisiana were monitored to judge what type of parental involvement yielded the highest on-task behaviors among participants. 39 children ages 3 to 5 years participated in 3- or 4-inning baseball games with parents on the field. In the Parent on Bases League, a parent stood on first, second, and third base only. In the Parent with Every Child League, a parent stood next to every child on the field. Children were observed on the field at three different dates at 1-min. intervals during the first two innings of one game in both leagues. On-task behaviors were defined as standing in the correct field position and watching the ball. Off-task behaviors consisted of sitting, rolling on the field, talking to a coach or parent, and not watching the ball. A statistically significant difference in the percent of time children spent on-task favored the Parents on Bases League (75.7) versus the other league (50.3). Having fewer parents on the field in these baseball games kept players on task more effectively than having a parent with each child. PMID- 17918566 TI - Comparing children with and without dyslexia on the Movement Assessment Battery for Children and the Test of Gross Motor Development. AB - Dyslexia is the most commonly occurring learning disability in the United States, characterized by difficulties with word recognition, spelling, and decoding. A growing body of literature suggests that deficits in motor skill performance exist in the dyslexic population. This study compared the performance of children with and without dyslexia on different subtests of the Test of Gross Motor Development and Movement Assessment Battery for Children and assessed whether there were developmental changes in the scores of the dyslexic group. Participants included 26 dyslexic children (19 boys and 7 girls; 9.5 yr. old, SD = 1.7) and 23 age- and sex-matched typically developing (17 boys and 6 girls; 9.9 yr. old, SD = 1.3) children as a control group. Mann-Whitney U tests indicated that the dyslexic group performed significantly lower than the control group only on the Total Balance subtest of the Movement Assessment Battery for Children. Additionally, the young dyslexic group performed significantly better on the Total Balance subtest, compared to the older dyslexic group. These results suggest that cerebellar dysfunction may account for differences in performance. PMID- 17918567 TI - Correlations between speakers' body size and acoustic parameters of voice. AB - Generally, there is a significant relationship between some acoustic measures (F0 and formant parameters) and the body size of speakers; however, data become less clear when age and sex variables are controlled. To date, no other vocal parameter apart from F0 has been studied in relation to body size. In the present study, correlations between a set of 27 parameters of the Multi-dimensional Voice Program (Kay Elemetrics Corp.) and 4 body measures were obtained from 134 speakers of both sexes belonging to one age group (20-29 years). Correlations within sex groups were null or very weak, and all significant coefficients were below .35. PMID- 17918568 TI - Relations of age with changes in self-efficacy and physical self-concept in preadolescents participating in a physical activity intervention during afterschool care. AB - An inference from Piaget's theory of cognitive development suggests that preadolescents of different ages who participate in a physical activity intervention may respond differently on measures of their physical self and self efficacy, so a field investigation was conducted to assess effects. In a sample of 105 children ages 8 to 12 yr. (42% boys, overall M(age) = 10.1 yr., SD = 0.9), participation in a physical activity intervention during afterschool care, based on social cognitive theory and incorporating instruction in self-management and self-regulatory skills, was associated with significant improvements in measures of exercise-related self-efficacy, perceived physical appearance, and physical self-concept over 12 wk. Analyses suggested, however, no difference in changes on these factors was associated with participants' age or children being in either the concrete operations or formal operations stage of cognitive development. After replication, implications for design of physical activity interventions for preadolescents were suggested. PMID- 17918569 TI - Self-perception of health and fitness among French and Tunisian men and women. AB - This work examines the effects of sex and culture on physical self-perceptions. The aim was to compare the perception of physical fitness of French and Tunisian men and women. 400 individuals ages 20 to 35 years assessed their own fitness, endurance, strength, flexibility, body composition, and health according to specific category scales by completing a questionnaire. In general, the Tunisian group rated themselves higher than the French group. It appears that perceived physical fitness was related mainly to perceived endurance for both groups. Some disparities were observed between the two nationalities. Analysis showed an interaction between sex and culture (French vs Tunisian). For French men and women and Tunisian men, perceived physical fitness was more associated with perceived endurance, whereas for Tunisian women, perceived physical fitness was more strongly associated with flexibility. These data show that self-perception of physical fitness is a dimension which varies between individuals from different cultures. PMID- 17918570 TI - Differential skills of perception of frequency. AB - This study aimed to develop tests for differentiating auditory perception skills, e.g., to test drug effects. In Exp. 1 (N = 60), discrimination of succeeding frequencies, frequency identification assessed by a choice reaction task, and the perception of pitch contour assessed by frequency deviants in binaural melodies showed zero partial correlations. In Exp. 2 (N = 36), discrimination correlated with performance on a field dependence test; fast frequency identification correlated with verbal fluency. The auditory tests are likely suitable for assessing distinct skills, but correlations with general cognitive abilities require further investigations. PMID- 17918572 TI - Intellectual disabilities and developmental EEG transition to alpha band: a comment on Shibagaki, et al. (2006). AB - In 2006 Shibagaki, et al. estimated mean chronological age at which dominant EEG frequency reached alpha band in 11 children with intellectual disabilities (M age, 15 yr.). Consistent with previous studies, the results showed a delay in the critical age relative to healthy children and earlier maturation at posterior scalp sites, with the exception of several children who persistently showed EEG slower than alpha band. A flaw in the study might be that having only 2 children younger than 10 yr. was insufficient. A remaining problem for researchers would be to identify whether the developmental delay in EEG correlates with IQ in intellectually disabled children. PMID- 17918571 TI - Performance dissociation during verbal and spatial working memory tasks. AB - Past research has inconsistently distinguished the neural substrates of various types of working memory. Task design and individual performance differences are known to alter patterns of brain response during working-memory tasks. These task and individual differences may have produced discrepancies in imaging findings. This study of 50 healthy adults (M(age) = 19.6 yr., SD = .8) examined performance during various parametric manipulations of a verbal and spatial n-back working memory task. Performance systematically dissociated on the basis of working memory load, working memory type, and stimulus difficulty, with participants having greater accuracy but slower response time during conditions requiring verbal versus spatial working memory. These findings hold implications for cognitive and neuroimaging studies of verbal and spatial working memory and highlight the importance of considering both task design and individual behavior. PMID- 17918573 TI - Effects of contour proximity and lightness on Delboeuf illusions created by circumscribed letters. AB - 32 observers judged the size of a letter, either an "A" or an "S," which was surrounded by a circle. Both letters were overestimated, but larger surrounding circles reduced the illusion. Decreasing the lightness contrast of the surrounding circle relative to the central letter diminished the illusion. The results suggest that, like the Delboeuf illusion, these circumscribed letters illusions are produced by interactions among size-coding neurons. PMID- 17918574 TI - Correlations of conceptual development with motor skills for a Turkish sample of kindergarten children. AB - The purpose was to research the conceptual development and motor skills of 36 healthy children of kindergarten age (M age = 67.6 mo., SD = 3.6). 19 girls and 17 boys completed the Bracken Basic Concept Scale-Revised and the Bruininks Oseretsky Test of Motor Proficiency-Short Form. PMID- 17918575 TI - Extraversion and neuroticism in team sport participants, individual sport participants, and nonparticipants. AB - Scores on Extraversion and on Neuroticism as measured by the Eysenck Personality Inventory were compared for 90 undergraduate team sport participants, individual sport participants, and nonparticipants (43 men, 47 women, M age = 20.3 yr.). From past research and Eysenck's biological theory of personality, it was hypothesized that sport participants would score higher on Extraversion and lower on Neuroticism than nonparticipants, and that team participants would score higher on Extraversion and perhaps higher on Neuroticism than individual sport participants. By comparing scores for students in first year and final year, it was also investigated whether pre-existing personality differences drew people to sport (the gravitational hypothesis) or whether personality changed as a function of sport participation (the developmental hypothesis). The main findings were that team participants scored higher on Extraversion than both individual sport participants and nonparticipants, and that test scores did not change over time, supporting the gravitational hypothesis for Extraversion. PMID- 17918576 TI - Lack of degradation in visuospatial perception of line orientation after one night of sleep loss. AB - Sleep deprivation impairs a variety of cognitive abilities including vigilance, attention, and executive function. Although sleep loss has been shown to impair tasks requiring visual attention and spatial perception, it is not clear whether these deficits are exclusively a function of reduced attention and vigilance or if there are also alterations in visuospatial perception. Visuospatial perception and sustained vigilance performance were therefore examined in 54 healthy volunteers at rested baseline and again after one night of sleep deprivation using the Judgment of Line Orientation Test and a computerized test of psychomotor vigilance. Whereas psychomotor vigilance declined significantly from baseline to sleep-deprived testing, scores on the Judgment of Line Orientation did not change significantly. Results suggest that documented performance deficits associated with sleep loss are unlikely to be the result of dysfunction within systems of the brain responsible for simple visuospatial perception and processing of line angles. PMID- 17918577 TI - Complexity of visual icons studied via signal detection theory. AB - Two investigations on how humans perceive information from visually rendered complex objects, such as military icons (glyphs) were conducted. A signal detection theory framework was employed to evaluate the sensitivity and specificity of human subject performance. The 6 adults tested showed that as complexity increased, their accuracy in performance decreased. Study 1 showed that complex dimensions (features) could not be assigned arbitrarily. Study 2 developed a rank ordering for features of an iconic object. PMID- 17918578 TI - Influence of attention manipulation on emotion and autonomic responses. AB - Psychopathologies such as depression and anxiety have been associated with self consciousness, a trait focusing on the self in terms of emotions and social images. A technique designed to shift attention away from the self tends to reduce anxiety, so the present purpose was to assess the effect of self body state information on an individual's emotional and autonomic activity. 24 undergraduate and graduate students (10 men and 14 women), ages 19 to 27 years (M = 22.1, SD = 2.5), were recruited as subjects. Focusing on body-state during an anxiety-inducing situation led to an increase of low to high frequency ratio of heart-rate variability which reflected cardiac sympathovagal balance. That is, attending to one's own bodily states enhanced relative sympathetic activity compared to parasympathetic activity, which can be interpreted as one of the physiological emotional responses elicited by anxiety. PMID- 17918579 TI - Evaluating the factor structure of the Psychological Performance Inventory. AB - This study assesses the construct validity of a measure of mental toughness, Loehr's Psychological Performance Inventory. Performers (N = 408, 303 men, 105 women, M age = 24.0 yr., SD = 6.7) drawn from eight sports (artistic rollerskating, basketball, canoeing, golf, rugby league, rugby union, soccer, swimming), and competing at either international, national, county and provincial, or club and regional standards. They completed the 42-item Psychological Performance Inventory during training camps. Principal components analysis provided minimal support for the factor structure. Instead, the exploratory analysis yielded a 4-factor 14-item model (PPI-A). A single factor underlying mental toughness (G(MT)) was identified with higher-order exploratory factor analysis using the Schmid-Leiman procedure. Psychometric analysis of the model, using confirmatory analysis techniques, fitted the data well. Collectively satisfying absolute and incremental fit index benchmarks, the inventory possesses satisfactory psychometric properties, with adequate reliability and convergent and discriminant validity. The results lend preliminary support to the factorial validity and reliability of the model; however, further investigation of its stability is required before recommending practitioners use changes in scores as an index for evaluating effects of training in psychological skills. PMID- 17918580 TI - An empirical test of sex differences in the emphasis on physical attractiveness in mate selection. AB - Within a context provided by social structural theory, social evolutionary theory, and physical attractiveness stereotyping, the importance of physical attractiveness in heterosexual mate selection was explored by presenting 50 male and 50 female psychology students (M age = 22.5 yr.) during a scheduled class with an opposite sex personals advertisement, wherein the advertiser was described as 'average' or 'good-looking'. Dependent variables consisted of a written paragraph and measures of evaluation (Semantic Differential), attraction, advertisement appeal, and success. An interaction for sex x looks on the qualitative measure showed no effect for men, but the good-looking female advertiser was evaluated more positively. However, for quantitative data, the advertisement was seen as more appealing and likely to be successful when the advertiser was good looking as opposed to average looking, irrespective of sex of advertiser. Findings are discussed in relation to theoretical perspectives. PMID- 17918581 TI - Precompetitive anxiety and self-confidence in athletes with disability. AB - This study examined the precompetition temporal patterning of competitive anxiety components in 42 athletes with disability who participated at the national level and at the national trials for the Paralympic Games in a variety of sports. All subjects completed a modified version of the Competitive State Anxiety Inventory 2 which measures intensity and direction of the competitive anxiety response on three occasions before competition (1 wk., 2 hr., and 20 min.). Analysis suggested that for cognitive and somatic dimensions athletes with disabilities show a similar precompetition anxiety response to athletes without disability. However, there appear to be some differences, particularly in the intensity of self-confidence, as athletes with disability reported a reduction of self confidence just prior to competition. PMID- 17918582 TI - Gastroesophageal reflux disease: alexithymia and attachment style. AB - Alexithymia and its relation with attachment style were evaluated in a group of 69 patients (men, M age = 46.4 yr., SD = 12.6; women, M age = 44.2 yr., SD = 14.4) affected by Gastroesophageal Reflux Disease. Two self-evaluation questionnaires were used for psychological evaluation, the 20 item Toronto Alexithymia Scale (TAS-20) and the Attachment Style Questionnaire (ASQ). The TAS 20 analysis showed that the clinical sample taken as a whole did not score in the alexithymic range. The inverse correlations between the Confidence ASQ subscale and the Difficulty Communicating Feelings TAS-20 subscale showed that communication of emotions could develop more easily within the framework of a relational context characterized by safety and confidence. In this group of patients this was represented by the referent caregiver. PMID- 17918583 TI - Is prism adaptation "for" growth? AB - The assumption that prism adaptation mechanisms evolved for developmental plasticity was questioned by analyzing natural transformations (magnification, rotation, displacement) of the arm and shoulder. Accommodating ordinary movement was found to be a closer match to prisms than transformations caused by growth. In addition, overlap between equations of movement and growth may point to a distal function of adaptation that is very general. PMID- 17918584 TI - Sludge reduction with Tubificidae and the impact on the performance of the wastewater treatment process. AB - To reduce excess sludge, a Tubificidae reactor was combined with an integrated oxidation ditch with vertical circle (IODVC), and a new integrated system was developed for wastewater treatment. A pilot-scale of this integrated system was tested to investigate the sludge reduction with Tubificidae and the impact on effluent quality and sludge production. The dominant worm was Branchnria Sowerbyi in the Tubificidae reactor after inoculation of Branchnria Sowerbyi and Limnodrilns sp., and the maximal volume density of wet Tubificidae in vessels of the Tubificidae reactor was 17600 g/m3. Two operational modes, treating the excess sludge (first mode) and the returned sludge (second mode) of IODVC by the Tubificidae reactor, were used in this experiment. The results showed that the excess sludge reduction rate was 46.4% in the first mode, and the average sludge yield of the integrated system was 6.19 x 10(-5) kg SS/kg COD in the second mode. Though the sludge returned to IODVC via the Tubificidae reactor, it had little impact on the effluent quality and the sludge characteristics of the IODVC. No new type of recalcitrant substance in the supernatant was discharged into the environment when the sludge was treated by Tubificidae. The experimental results also indicated that no significant changes occurred on the viscosity, specific resistance, and the floc size distribution of the sludge. PMID- 17918585 TI - Characteristics of high-sulfate wastewater treatment by two-phase anaerobic digestion process with Jet-loop anaerobic fluidized bed. AB - A new anaerobic reactor, Jet-loop anaerobic fluidized bed (JLAFB), was designed for treating high-sulfate wastewater. The treatment characteristics, including the effect of influent COD/SO4(2-) ratio and alkalinity and sulfide inhibition in reactors, were discussed for a JLAFB and a general anaerobic fluidized bed (AFB) reactor used as sulfate-reducing phase and methane-producing phase, respectively, in two-phase anaerobic digestion process. The formation of granules in the two reactors was also examined. The results indicated that COD and sulfate removal had different demand of influent COD/SO4(2-) ratios. When total COD removal was up to 85%, the ratio was only required up to 1.2, whereas, total sulfate removal up to 95% required it exceeding 3.0. The alkalinity in the two reactors increased linearly with the growth of influent alkalinity. Moreover, the change of influent alkalinity had no significant effect on pH and volatile fatty acids (VFA) in the two reactors. Influent alkalinity kept at 400-500 mg/L could meet the requirement of the treating process. The JLAFB reactor had great advantage in avoiding sulfide and free-H2S accumulation and toxicity inhibition on microorganisms. When sulfate loading rate was up to 8.1 kg/(m3 x d), the sulfide and free-H2S concentrations in JLAFB reactor were 58.6 and 49.7 mg/L, respectively. Furthermore, the granules, with offwhite color, ellipse shape and diameters of 1.0-3.0 mm, could be developed in JLAFB reactor. In granules, different groups of bacteria were distributed in different layers, and some inorganic metal compounds such as Fe, Ca, Mg etc. were found. PMID- 17918587 TI - Effect of coagulation pretreatment on the fouling of ultrafiltration membrane. AB - The purpose of this study is to understand the effect and mechanism of preventing membrane fouling, by coagulation pretreatment, in terms of fractional component and molecular weight of natural organic matter (NOM). A relatively higher molecular weight (MW) of hydrophobic compounds was responsible for a rapid decline in the ultrafiltration flux. Coagulation could effectively remove the hydrophobic organics, resulting in the increase of flux. It was found that a lower MW of neutral hydrophilic compounds, which could remove inadequately by coagulation, was responsible for the slow declining flux. The fluxes in the filtration of coagulated water and supernatant water were compared and the results showed that a lower MW of neutral hydrophilic compounds remained in the supernatant water after coagulation could be rejected by a membrane, resulting in fouling. It was also found that the coagulated flocs could absorb neutral hydrophilic compounds effectively. Therefore, with the coagulated flocs formed on the membrane surface, the flux decline could be improved. PMID- 17918586 TI - Transformations of particles, metal elements and natural organic matter in different water treatment processes. AB - Characterizing natural organic matter (NOM), particles and elements in different water treatment processes can give a useful information to optimize water treatment operations. In this article, transformations of particles, metal elements and NOM in a pilot-scale water treatment plant were investigated by laser light granularity system, particle counter, glass-fiber membrane filtration, inductively coupled plasma-optical emission spectroscopy, ultra filtration and resin absorbents fractionation. The results showed that particles, NOM and trihalomethane formation precursors were removed synergistically by sequential treatment of different processes. Pre-ozonation markedly changed the polarity and molecular weight of NOM, and it could be conducive to the following coagulation process through destabilizing particles and colloids; mid-ozonation enhanced the subsequent granular activated carbon (GAC) filtration process by decreasing molecular weight of organic matters. Coagulation-flotation and GAC were more efficient in removing fixed suspended solids and larger particles; while sand-filtration was more efficient in removing volatile suspended solids and smaller particles. Flotation performed better than sedimentation in terms of particle and NOM removal. The type of coagulant could greatly affect the performance of coagulation-flotation. Pre-hydrolyzed composite coagulant (HPAC) was superior to FeCl3 concerning the removals of hydrophobic dissolved organic carbon and volatile suspended solids. The leakages of flocs from sand-filtration and microorganisms from GAC should be mitigated to ensure the reliability of the whole treatment system. PMID- 17918588 TI - Denitrification potential enhancement by addition of external carbon sources in a pre-denitrification process. AB - The aim of this study is to investigate the denitrification potential enhancement by addition of external carbon sources and to estimate the denitrification potential for the predenitrification system using nitrate utilization rate (NUR) batch tests. It is shown that the denitrification potential can be substantially increased with the addition of three external carbon sources, i.e. methanol, ethanol, and acetate, and the denitrification rates of ethanol, acetate, and methanol reached up to 9.6, 12, and 3.2 mgN/(g VSS x h), respectively, while that of starch wastewater was only 0.74 mgN/(g VSS x h). By comparison, ethanol was found to be the best external carbon source. NUR batch tests with starch wastewater and waste ethanol were carried out. The denitrification potential increased from 5.6 to 16.5 mg NO3-N/L owing to waste ethanol addition. By means of NUR tests, the wastewater characteristics and kinetic parameters can be estimated, which are used to determine the denitrification potential of wastewater, to calculate the denitrification potential of the plant and to predict the nitrate effluent quality, as well as provide information for developing carbon dosage control strategy. PMID- 17918589 TI - Biological control experiment of excess propagation of Cyclops for drinking water security. AB - Cyclops of zooplankton propagated excessively in eutrophic water body and could not be effectively inactivated by the conventional disinfections process like chlorination due to its stronger resistance to oxidation. In this study, an ecological project was put forward for the excess propagation control of Cyclops by stocking the filter-feeding fishes such as silver carp and bighead carp under the condition of no extraneous nutrient feeding. The results of experiments with different stocking biomass showed that the propagation of Cyclops could be controlled effectively, and the water quality was improved simultaneously by impacting on nutriment level and plankton community structure at proper stocking density of 30 g/m3 of water. The growth of Cyclops may not be effectually controlled with lower biomass of fish (10 g), and the natural food chain relation may be destroyed for Cyclops dying out in water while the intense stocking of 120 g per cubic meter of water. In addition, the high predator pressure may accelerate supplemental rate of nutrients from bottom sediments to water body to add the content of total nitrogen and phosphorus in water. PMID- 17918590 TI - First flush of storm runoff pollution from an urban catchment in China. AB - Storm runoff pollution process was investigated in an urban catchment with an area of 1.3 km2 in Wuhan City of China. The results indicate that the pollutant concentration peaks preceded the flow peaks in all of 8 monitored storm events. The intervals between pollution peak and flow peak were shorter in the rain events with higher intensity in the initial period than those with lower intensity. The fractions of pollution load transported by the first 30% of runoff volume (FF30) were 52.2%-72.1% for total suspended solids (TSS), 53.0%-65.3% for chemical oxygen demand (COD), 40.4%-50.6% for total nitrogen (TN), and 45.8% 63.2% for total phosphorus (TP), respectively. Runoff pollution was positively related to non-raining days before the rainfall. Intercepting the first 30% of runoff volume can remove 62.4% of TSS load, 59.4% of COD load, 46.8% of TN load, and 54.1% of TP load, respectively, according to all the storm events. It is suggested that controlling the first flush is a critical measure in reduction of urban stormwater pollution. PMID- 17918591 TI - Chemical oxygen demand reduction in coffee wastewater through chemical flocculation and advanced oxidation processes. AB - The removal of the natural organic matter present in coffee processing wastewater through chemical coagulation-flocculation and advanced oxidation processes (AOP) had been studied. The effectiveness of the removal of natural organic matter using commercial flocculants and UV/H2O2, UV/O3 and UV/H2O2/O3 processes was determined under acidic conditions. For each of these processes, different operational conditions were explored to optimize the treatment efficiency of the coffee wastewater. Coffee wastewater is characterized by a high chemical oxygen demand (COD) and low total suspended solids. The outcomes of coffee wastewater treatment using coagulation-flocculation and photodegradation processes were assessed in terms of reduction of COD, color, and turbidity. It was found that a reduction in COD of 67% could be realized when the coffee wastewater was treated by chemical coagulation-flocculation with lime and coagulant T-1. When coffee wastewater was treated by coagulation-flocculation in combination with UV/H2O2, a COD reduction of 86% was achieved, although only after prolonged UV irradiation. Of the three advanced oxidation processes considered, UV/H2O2, UV/O3 and UV/H2O2/O3, we found that the treatment with UV/H2O2/O3 was the most effective, with an efficiency of color, turbidity and further COD removal of 87%, when applied to the flocculated coffee wastewater. PMID- 17918592 TI - Characterization and distribution of polycyclic aromatic hydrocarbon in sediments of Haihe River, Tianjin, China. AB - In this study sediment samples were collected from 13 sites of Haihe River in Tianjin City, China, sixteen of priority polycyclic aromatic hydrocarbons (PAHs) listed in USEPA were analyzed by means of GC-MS. The total concentrations of PAH ranged from 774.81 to 255371.91 ng/g dw, and two to four rings of PAHs were dominant in sediment samples. Molecular ratios, such as phenanthrene/anthracene, fluoranthene/pyrene and low-molecular-weight PAH /high-molecular-weight PAH, were used to study the possible sources of pollution. It indicated a mixed pattern of parolytic and petrogenic inputs of PAHs in sediments in Haihe River. The petrogenic PAHs may be mainly derived from the leakage of refined products, e.g., gasoline, diesel fuel and fuel oil vehicle traffics or gas stations from urban area. The pyrolytic PAHs might be from the discharge of industrial wastewater and the emission of atmospheric particles from petrochemical factories. In addition, the levels of PAHs in the urban and industrial areas are far beyond the values reported from other rivers and marine systems reported. This situation may be due to polluted discharging from some petrochemical industrial manufactories and worse traffic conditions in Tianjin. PMID- 17918593 TI - Adsorption behavior of condensed phosphate on aluminum hydroxide. AB - Sodium pyrophosphate (pyro-P, Na4P2O7), sodium tripolyphosphate (tripoly-P, Na5P3O10), and sodium hexametaphosphate (meta-P, (NaPO3)6) were selected as the model compounds of condensed phosphate to investigate the adsorption behavior of condensed phosphate on aluminum hydroxide. The adsorption was found to be endothermic and divisible into two stages: (1) fast adsorption within 1 h; and (2) slow adsorption between 1 and 24 h. The modified Freundlich model simulated the fast adsorption stage well; the slow adsorption stage was described well by the first-order kinetics. The activation energies of pyro-P, tripoly-P, and meta P adsorption on aluminum hydroxide were determined to be 20.2, 22.8 and 10.9 kJ/mol P adsorbed, respectively, in the fast adsorption stage and to be 66.3, 53.5 and 72.5 kJ/mol P adsorbed, respectively, in the slow adsorption stage. The adsorption increased the negative charge of the aluminum hydroxide surface. Transmission electron microscopy and energy dispersive X-ray analysis analyses provided evidence that the adsorption was not uniform on the surface and that the small crystals contributed more to the fast adsorption than the normal sites did. The results from X-ray fluorescence spectrometry and X-ray photoelectron spectroscopy tests also revealed the uneven adsorption of condensed phosphate as a function of the penetration depth. More condensed phosphates were adsorbed on the outer surface of aluminum hydroxide than in its inner parts. PMID- 17918594 TI - Topsoil organic carbon mineralization and CO2 evolution of three paddy soils from South China and the temperature dependence. AB - Carbon mineralization and its response to climatic warming have been receiving global attention for the last decade. Although the virtual influence of temperature effect is still in great debate, little is known on the mineralization of organic carbon (SOC) of paddy soils of China under warming. SOC mineralization of three major types of China's paddy soils is studied through laboratory incubation for 114 d under soil moisture regime of 70% water holding capacity at 20 degrees C and 25 degrees C respectively. The carbon that mineralized as CO2 evolved was measured every day in the first 32 d and every two days in the following days. Carbon mineralized during the 114 d incubation ranged from 3.51 to 9.22 mg CO2-C/gC at 20 degrees C and from 4.24 to 11.35 mg CO2-C/gC at 25 degrees C respectively; and a mineralizable C pool in the range of 0.24 to 0.59 gC/kg, varying with different soils. The whole course of C mineralization in the 114 d incubation could be divided into three stages of varying rates, representing the three subpools of the total mineralizable C: very actively mineralized C at 1-23 d, actively mineralized C at 24-74 d and a slowly mineralized pool with low and more or less stabilized C mineralization rate at 75 114 d. The calculated Q10 values ranged from 1.0 to 2.4, varying with the soil types and N status. Neither the total SOC pool nor the labile C pool could account for the total mineralization potential of the soils studied, despite a well correlation of labile C with the shortly and actively mineralized C, which were shown in sensitive response to soil warming. However, the portion of microbial C pool and the soil C/N ratio controlled the C mineralization and the temperature dependence. Therefore, C sequestration may not result in an increase of C mineralization proportionally. The relative control of C bioavailability and microbial metabolic activity on C mineralization with respect to stabilization of sequestered C in the paddy soils of China is to be further studied. PMID- 17918595 TI - Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents. AB - Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter. PMID- 17918596 TI - Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria. AB - A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37 degrees C. It can utilize several (halo)aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30 degrees C with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no (chloro)catechol 2,3-dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway. PMID- 17918597 TI - Occurrence and congeners specific of polychlorinated biphenyls in agricultural soils from Southern Jiangsu, China. AB - A total of 198 agricultural soil samples were collected from Zhangjiagang and Changshu in Southern Jiangsu for analysis of 13 polychlorinated biphenyls (PCBs) in order to assess the levels of pollution, sources, area distribution, and potential risk for the environment. All methods were rigorously tested and an adequate quality control was ensured. Only one site had no PCBs residues, and the highest total PCBs concentration in the surface soils was 32.83 ng/g. The average concentration in all the soil samples was 4.13 ng/g, signaling low-level pollution. Tetra-, penta-, and hexa-chlorinated biphenyls were dominant species in soil samples, accounting for more than 75% of sigmaPCBs in the soil samples. PCB118 was the most abundant congener in all the samples. The PCB118 was about 20% of sigmaPCBs. The soil organic matter content showed only a weak correlation with the levels of all PCB congeners, in which a better correlation was noted for the more volatile lighter PCB congeners than for the heavier homologues. To a certain extent, the sources and land use seemed to influence the levels of PCBs. PMID- 17918598 TI - Fractionation and solubility of cadmium in paddy soils amended with porous hydrated calcium silicate. AB - Previous studies have shown that porous hydrated calcium silicate (PS) is very effective in decreasing cadmium (Cd) content in brown rice. However, it is unclear whether the PS influences cadmium transformation in soil. The present study examined the effect of PS on pH, cadmium transformation and cadmium solubility in Andosol and Alluvial soil, and also compared its effects with CaCO3, acidic porous hydrated calcium silicate (APS) and silica gel. Soil cadmium was operationally fractionationed into exchangeable (Exch), bound to carbonates (Carb), bound to iron and manganese oxides (FeMnO(x)), bound to organic matters (OM) and residual (Res) fraction. Application of PS and CaCO3 at hig rates enhanced soil pH, while APS and silica gel did not obviously change soil pH. PS and CaCO3 also increased the FeMnO(x)-Cd in Andosol and Carb-Cd in Alluvial soil, thus reducing the Exch-Cd in the tested soils. However, PS was less effective than CaCO3 at the same application rate. Cadmium fractions in the two soils were not changed by the treatments of APS and silica gel. There were no obvious differences in the solubility of cadmium in soils treated with PS, APS, silica gel and CaCO3 except Andosol treated 2.0% CaCO3 at the same pH of soil-CaCl2 suspensions. These findings suggested that the decrease of cadmium availability in soil was mainly attributed to the increase of soil pH caused by PS. PMID- 17918599 TI - Variation of soil fertility and carbon sequestration by planting Hevea brasiliensis in Hainan Island, China. AB - The development of rubber industry depends on the sustainable management of rubber plantation. To evaluate the environmental effects of planting Hevea brasiliensis on a subsystem of tropical forest ecosystem, the variation of soil fertility and carbon sequestration under rubber plantation within 30-year life period were investigated in Hainan Island. Results showed that (1) with the increase of stand age of rubber plantation, soil fertility decreased all along. From 1954 to 1995, soil organic matter, total N, available K and available P decreased by 48.2%, 54.1%, 56.7% and 64.1%, respectively. (2) If the complete return of litters was considered without additional fertilizer application to the soil of the rubber plantations, the consumption periods for P, N, K, Mg were only 825 years, 329 years, 94 years and 65 years, respectively. To improve soil fertility is essential for rubber plantation development. (3) The C sequestration of rubber trees per hectare accounts for 272.08 t within 30-year life period and 57.91% of them was fixed in litters. In comparison with C sequestration by rain forest (234.305 t/hm2) and by secondary rain forest (150.203 t/hm2), rubber forest has more potentials for C fixation. On the base of above results, the following measures would benefit the maintenance of soil fertility and the development of rubber industry, including applying fertilizer to maintain the balance of soil nutrients, intercropping leguminous plant to improve soil fertility, reducing the collection of litters, optimizing soil properties to improve element P availability such as applying CaCO3. The information gathered from the study can be used as baseline data for the sustainable management of rubber plantation elsewhere. PMID- 17918600 TI - Vertical distribution of nonylphenol and nonylphenol ethoxylates in sedimentary core from the Beipaiming Channel, North China. AB - The vertical profiles of nonylphenol (NP) and nonylphenol ethoxylates (NPEOs) were investigated in a sediment core from the Beipaiming Channel, North China using high sensitive LC-MS and GC-MS methods. In this core, relatively high concentrations of NP and NPEOs occurred in the surface sediment (< or = 40 cm), with the maximum value of NP and NPEOs reaching 3539 and 12735 ng/g, respectively, whereas, no NP or NPEOs were detected in deeper sediments (> 40 cm). The high concentrations of NP and NPEOs in the surface layers suggested recent inputs in this area. NPEOs with short ethoxy chains (NPnEO, n = 0-3) were dominant in the NPEO mixture with percentages from 54% to 78%, which were similar to the distribution of homolog NPEO in effluents from nearby sewage treatment plants (STPs), indicating that the channel received the effluents from these STPs. The sewage treatment ratio was quite similar to that found in North America before the 1980s. Finally, the concentrations of NP and NPEOs were related to the total organic carbon (TOC) (p < 0.001), suggesting that TOC was an important factor for vertical distribution of NPEOs and NP from the Beipaiming Channel. PMID- 17918601 TI - Photocatalytic remediation of gamma-hexachlorocyclohexane contaminated soils using TiO2 and montmorillonite composite photocatalyst. AB - TiO2 and montmorillonite composite photocatalysts were prepared and applied in degrading gamma-hexachlorocyclohexane (gamma-HCH) in soils. After being spiked with gamma-HCH, soil samples loaded with the composite photocatalysts were exposed to UV-light irradiation. The results indicated that the photocatalytic activities of the composite photocatalysts varied with the content of TiO2 in the order of 10% < 70% < 50% < 30%. Moreover, the photocatalytic activity of the composite photocatalysts with TiO2 content 30% was higher than that of the pure P25 with the same mass of TiO2. The strong adsorption capacity of the composite photocatalysts and quantum size effect may contribute to its increased photocatalytic activities. In addition, effect of dosage of composite photocatalysts and soil pH on gamma-HCH photodegradation was investigated. Pentachlorocyclohexene, trichlorocyclohexene, and dichlorobenzene were detected as photodegradation intermediates, which were gradually degraded with the photodegradation evolution. PMID- 17918602 TI - Catalytic reductive dechlorination of p-chlorophenol in water using Ni/Fe nanoscale particles. AB - Nanoscale bimetallic Ni/Fe particles were synthesized from the reaction of sodium borohydride (NaBH4) with reduction of Ni2+ and Fe2+ in aqueous solution. The obtained Ni/Fe particles were characterized by TEM (transmission electron microscope), XRD (X-ray diffractometer), and N2-BET. The dechlorination activity of the Ni/Fe was investigated using p-chlorophenol (p-CP) as a probe agent. Results demonstrated that the nanoscale Ni/Fe could effectively dechlorinate p-CP at relatively low metal to solution ratio of 0.4 g/L (Ni 5 wt%). The target with initial concentration of p-CP 0.625 mmol/L was dechlorinted completely in 60 min under ambient temperature and pressure. Factors affecting dechlorination efficiency, including reaction temperature, pH, Ni loading percentage over Fe, and metal to solution ratio, were investigated. The possible mechanism of dechlorination of p-CP was proposed and discussed. The pseudo-first-order reaction took place on the surface of the Ni/Fe bimetallic particles, and the activation energy of the dechlorination reaction was determined to be 21.2 kJ/mol at the temperature rang of 287-313 K. PMID- 17918603 TI - A new photon kinetic-measurement based on the kinetics of electron-hole pairs in photodegradation of textile wastewater using the UV-H2O2FS-TiO2 process. AB - Actual textile wastewater and synthesized wastewater containing various textile dyes were photocatalytic degraded by the UV-H2O2FS-TiO2 process in an annular flow photocatalytic reactor. In this process, a photon kinetic-measure was adopted to obtain constant rates of dyes decomposition. It was theorized that, by illumination at different UV frequencies, the electrons within the semiconductor were excited from the valence band to the conduction band, yielding the formation of electron-hole pairs which are the pre-requisites for photocatalysis. CPT (critical photonic time) exposure required to cause 90% of vibrations between the double and single bonds along the molecular chain of the dyes to be oxidized, was taken to measure the photocatalytic activities. The CPTs varied with the frequencies of the UV spectral areas. The derivatization of CPT from the first order kinetic law was presented. PMID- 17918604 TI - Bioconcentration kinetics of PCBs in various parts of the lifecycle of the tadpoles Xenopus laevis. AB - Polychlorinated biphenyls (PCBs) in Xenopus laevis have been reported only for a few congeners. Additionally, there is very little information on the ability of Xenopus laevis to bioconcentrate PCBs. To address these issues, the tadpole Xenopus laevis was exposed to Aroclor1254 mixtures in water at room temperature for 110 d followed by an additional 110 d of nonspiked PCBs in the water for the control group. During the whole process, bioconcentration factors (BCFs) of PCBs ranged from 1180 to 15670. For most PCB congeners, the highest and lowest bioconcentrations of the kinetic curves were found to be remarkably simultaneous, respectively. All 141 PCB congeners under the same experimental conditions had no linear correlation on the lgBCF versus lgK(ow) relationship. The relationship between lgBCFs and lgK(ow) followed a parabolic pattern indicative of selective bioconcentration, suggesting that the kinetic curves of the PCB congeners observed in the lifecycle of the tadpoles may be concentrated due to the amphibian special species and internal metabolism. In contrast, lgBCFs for PCBs were inversely related to lgK(ow), suggesting that a metabolism of the higher K(ow)' PCB congeners occurred. These results support the author's conclusion that the tadpole Xenopus laevis plays major roles in the bioconcentration of PCB congeners, and demonstrated that the exposure kinetic curves of PCB congeners are complex. Besides the amphibian metamorphous development, the lifecycle of the tadpole Xenopus laevis also may be of importance in determining the bioconcentration of PCB congeners. PMID- 17918605 TI - [Interdisciplinary consultation with nursing experts in delayed wound healing--2: Vacuum sealing in heart surgery]. PMID- 17918606 TI - [Problems from general practice--solutions for general practice: thrombosis prevention with compression stockings]. PMID- 17918607 TI - [Prevention of infection by vaccination: simple and effective protection]. PMID- 17918608 TI - [New (shared) responsibilities in health care occupations: assuring family health -a health care field for nursing]. PMID- 17918609 TI - [10 years Promotional Association to establish a Nursing Office in North Rhine Westphalia: supporting nursing self management]. PMID- 17918610 TI - [Educational pathways in nursing: qualifications for the future]. PMID- 17918611 TI - [Credit for extracurricular acquired competences in study courses: expanding educational perspectives, smoothing career paths]. PMID- 17918612 TI - [Spiritual impulses in the everyday routine of demented residents--educational project: breathing room for the soul]. PMID- 17918613 TI - [Observation task in practical education in geriatric psychiatry: developing interactive competence for person centered nursing]. PMID- 17918614 TI - [Step by step to nursing care competence--5: Fluid therapy. Critical life support]. PMID- 17918616 TI - [Human images in nursing--6: The person--the nurse: connection is a life source]. PMID- 17918615 TI - [Accidental falls in homes for the aged and nursing homes: no responsibility for constant observation]. PMID- 17918617 TI - [From case to case: between dream and reality]. PMID- 17918618 TI - [Software supports participation of patients in nursing care: validating the nursing care recipient as subject]. PMID- 17918619 TI - Virulence genes and the evolution of host specificity in plant-pathogenic fungi. AB - In the fungal kingdom, the ability to cause disease in plants appears to have arisen multiple times during evolution. In many cases, the ability to infect particular plant species depends on specific genes that distinguish virulent fungi from their sometimes closely related nonvirulent relatives. These genes encode host-determining "virulence factors," including small, secreted proteins and enzymes involved in the synthesis of toxins. These virulence factors typically are involved in evolutionary arms races between plants and pathogens. We briefly summarize current knowledge of these virulence factors from several fungal species in terms of function, phylogenetic distribution, sequence variation, and genomic location. Second, we address some issues that are relevant to the evolution of virulence in fungi toward plants; in particular, horizontal gene transfer and the genomic organization of virulence genes. PMID- 17918620 TI - Structural implications of mutations in the pea SYM8 symbiosis gene, the DMI1 ortholog, encoding a predicted ion channel. AB - The Pisum sativum SYM8 gene plays an essential part in both rhizobial and mycorrhizal symbioses. Mutation of sym8 in the original type line R25 blocks nodulation, mycorrhization, and Nod-factor-induced calcium spiking, an early component of the nodulation signaling pathway. We describe four new sym8 alleles of pea, which fall into the same complementation group as R25. The sym8 mutants are phenotypically similar to Medicago truncatula dmi1 mutants and map to a syntenic location. We used sequence homology to isolate the pea ortholog of M. truncatula DMI1 and have shown that the cloned pea ortholog can complement a M. truncatula dmi1 mutant for nodulation. Each of the five pea sym8 mutants carries a mutation in the DMI1 ortholog, confirming that the pea SYM8 is the DMI1 ortholog. Based on predicted structural similarities with an archaebacterial ion channel, we propose that SYM8 forms a tetrameric calcium-gated channel of a predicted structure similar to the archaebacterial potassium channel but containing a filter region that is different. The predicted structure identifies four aspartate residues (one from each subunit) forming the channel opening. We made a mutation changing the aspartate to valine and identified a missense mutation (changing alanine to valine adjacent to the aspartate residues) in this predicted filter region; both mutations caused a loss of function. We also identified a loss-of-function missense mutation (changing arginine to isoleucine) in a domain proposed to link the predicted channel and the gating ring domains, indicating that this mutation may block function by preventing a protein conformational change being transmitted from the gating-ring domain to the pore domain. PMID- 17918621 TI - A key role for the Arabidopsis WIN3 protein in disease resistance triggered by Pseudomonas syringae that secrete AvrRpt2. AB - Effector proteins injected by the pathogenic bacteria Pseudomonas syringae into plants can have profound effects on the pathogen-host interaction due to their efficient recognition by plants and the subsequent triggering of defenses. The AvrRpt2 effector triggers strong local and systemic defense (called systemic acquired resistance [SAR]) responses in Arabidopsis thaliana plants that harbor a functional RPS2 gene that encodes an R protein in the coiled-coil, nucleotide binding domain, leucine-rich repeat class. The newly identified win3-T mutant shows greatly reduced resistance to P syringae carrying avrRpt2. In win3-T plants, RIN4 cleavage, an early AvrRpt2-induced event, is normal. However, salicylic acid accumulation is compromised, as is SAR induction and the local hypersensitive cell death response after infection by P syringae carrying avrRpt2. WIN3 encodes a member of the firefly luciferase protein superfamily. Expression of WIN3 at an infection site partially requires PAD4, a protein known to play a quantitative role in RPS2-mediated signaling. WIN3 expression in tissue distal to an infection site requires multiple salicylic acid regulatory genes. Finally, win3-T plants show modestly increased susceptibility to virulent P syringae and modestly reduced SAR in response to P. syringae carrying avrRpm1. Thus, WIN3 is a key element of the RPS2 defense response pathway and a basal and systemic defense component. PMID- 17918622 TI - Agrobacterium VirD2-binding protein is involved in tumorigenesis and redundantly encoded in conjugative transfer gene clusters. AB - Agrobacterium tumefaciens can transfer oncogenic T-DNA into plant cells; T-DNA transfer is mechanistically similar to a conjugation process. VirD2 is the pilot protein that guides the transfer, because it is covalently associated with single stranded T-DNA to form the transfer substrate T-complex. We used the VirD2 protein as an affinity ligand to isolate VirD2-binding proteins (VBPs). By pull down assays and peptide-mass-fingerprint matching, we identified an A. tumefaciens protein designated VBP1 that could bind VirD2 directly. Genome-wide sequence analysis showed that A. tumefaciens has two additional genes encoding proteins highly similar to VBP1, designated vbp2 and vbp3. Like VBP1, both VBP2 and VBP3 also could bind VirD2; all three VBPs contain a putative nucleotidyltransferase motif. Mutational analysis of vbp demonstrated that the three vbp genes could functionally complement each other. Consequently, only inactivation of all three vbp genes highly attenuated the bacterial ability to cause tumors on plants. Although vbp1 is harbored on the megaplasmid pAtC58, vbp2 and vbp3 reside on the linear chromosome. The vbp genes are clustered with conjugative transfer genes, suggesting linkage between the conjugation and virulence factor. The three VBPs appear to contain C-terminal positively charged residues, often present in the transfer substrate proteins of type IV secretion systems. Inactivation of the three vbp genes did not affect the T-strand production. Our data indicate that VBP is a newly identified virulence factor that may affect the transfer process subsequent to T-DNA production. PMID- 17918623 TI - Antagonistic control of powdery mildew host cell entry by barley calcium dependent protein kinases (CDPKs). AB - Calcium-dependent protein kinases (CDPKs) are known to play pivotal roles in intracellular signaling during abiotic and biotic stress responses. To unravel potential functions of CDPKs in the course of barley (Hordeum vulgare)-powdery mildew (Blumeria graminis) interactions, we systematically analyzed the HvCDPK gene family. We found that, according to the existence of respective expressed sequence tags, at least nine paralogs are expressed in the barley leaf epidermis, the sole target tissue of powdery mildew fungi. We exemplarily selected two HvCDPKs with known full-length coding sequence for functional analysis. Transient expression of a putative constitutive active variant of one of these (HvCDPK4) in Nicotiana benthamiana triggered kinase-dependent mesophyll cell death in tobacco leaves. In a barley mlo mutant genotype, a constitutive active variant of the second paralog, HvCDPK3, partially compromised the highly effective resistance to B. graminis f. sp. hordei. A similar break of mlo resistance was seen upon expression of the junction domain of HvCDPK4, supposed to act as a dominant inhibitor of CDPK activity. Expression of a constitutive active HvCDPK3 or HvCDPK4 form also compromised penetration resistance to the inappropriate wheat powdery mildew fungus. Collectively, our data provide evidence for antagonistic roles of individual CDPK paralogs in the control of host cell entry during the early phase of powdery mildew pathogenesis. PMID- 17918624 TI - Biofilm formation, epiphytic fitness, and canker development in Xanthomonas axonopodis pv. citri. AB - The phytopathogenic bacterium Xanthomonas axonopodis pv. citri is responsible for the canker disease affecting citrus plants throughout the world. Here, we have evaluated the role of bacterial attachment and biofilm formation in leaf colonization during canker development on lemon leaves. Crystal violet staining and confocal laser scanning microscopy analysis of X. axonopodis pv. citri strains expressing the green fluorescent protein were used to evaluate attachment and biofilm formation on abiotic and biotic (leaf) surfaces. Wild-type X. axonopodis pv. citri attached to and formed a complex, structured biofilm on glass in minimal medium containing glucose. Similar attachment and structured biofilm formation also were seen on lemon leaves. An X. axonopodis pv. citri gumB mutant strain, defective in production of the extracellular polysaccharide xanthan, did not form a structured biofilm on either abiotic or biotic surfaces. In addition, the X. axonopodis pv. citri gumB showed reduced growth and survival on leaf surfaces and reduced disease symptoms. These findings suggest an important role for formation of biofilms in the epiphytic survival of X. axonopodis pv. citri prior to development of canker disease. PMID- 17918625 TI - Functional analysis of the metallothionein gene cgMT1 isolated from the actinorhizal tree Casuarina glauca. AB - cgMT1 is a metallothionein (MT)-like gene that was isolated from a cDNA library of young nitrogen-fixing nodules resulting from the symbiotic interaction between Frankia spp. and the actinorhizal tree Casuarina glauca. cgMT1 is highly transcribed in the lateral roots and nitrogen-fixing cells of actinorhizal nodules; it encodes a class I type 1 MT. To obtain insight into the function of cgMT1, we studied factors regulating the expression of the MT promoter region (PcgMT1) using a beta-glucuronidase (gus) fusion approach in transgenic plants of Arabidopsis thaliana. We found that copper, zinc, and cadmium ions had no significant effect on the regulation of PcgMT1-gus expression whereas wounding and H2O2 treatments led to an increase in reporter gene activity in transgenic leaves. Strong PcgMT1-gus expression also was observed when transgenic plants were inoculated with a virulent strain of the bacterial pathogen Xanthomonas campestris pv. campestris. Transgenic Arabidopsis plants expressing cgMT1 under the control of the constitutive 35S promoter were characterized by reduced accumulation of H2O2 when leaves were wounded and by increased susceptibility to the bacterial pathogen X. campestris. These results suggest that cgMT1 could play a role during the oxidative response linked to biotic and abiotic stresses. PMID- 17918626 TI - Novel reiterated Fnr-type proteins control the production of the symbiotic terminal oxidase cbb3 in Rhizobium etli CFN42. AB - Symbiotic nitrogen-fixing bacteria express a terminal oxidase with a high oxygen affinity, the cbb3-type oxidase encoded by the fixNOQP operon. Previously, we have shown that, in Rhizobium etli CFN42, the repeatedfixNOQP operons (fixNOQPd and fixNOQPf) have a differential role in nitrogen fixation. Only the fixNOQPd operon is required for the establishment of an effective symbiosis; microaerobic induction of this operon is under the control of at least three transcriptional regulators, FixKf, FnrNd, and FnrNchr, belonging to the Crp/Fnr family. In this work, we describe two novel Crp/Fnr-type transcriptional regulators (StoRd and StoRf, symbiotic terminal oxidase regulators) that play differential roles in the control of key genes for nitrogen fixation. Mutations either in stoRd or stoRf enhance the microaerobic expression of both fixNOQP reiterations, increasing also the synthesis of the cbb3-type oxidase in nodules. Despite their structural similarity, a differential role of these genes was also revealed, since a mutation in stoRd but not in stoRf enhanced both the expression of fixKf and the nitrogen-fixing capacity of R. etli CFN42. PMID- 17918627 TI - New type III effectors from Xanthomonas campestris pv. vesicatoria trigger plant reactions dependent on a conserved N-myristoylation motif. AB - Pathogenicity of the gram-negative plant pathogen Xanthomonas campestris pv. vesicatoria depends on a type III secretion system, which translocates bacterial effector proteins into the plant cell. In this study, we identified two novel type III effectors, XopE1 and XopE2 (Xanthomonas outer proteins), using the AvrBs3 effector domain as reporter. XopE1 and XopE2 belong to the HopX family and possess a conserved putative N-myristoylation motif that is also present in the effector XopJ from X. campestris pv. vesicatoria 85-10. XopJ is a member of the YopJ/AvrRxv family of acetyltransferases. Confocal laser scanning microscopy and immunocytochemistry revealed that green fluorescent protein fusions of XopE1, XopE2, and XopJ localized to the plant cell plasma membrane. Targeting to the membrane is probably due to N-myristoylation, because a point mutation in the putative myristoylated glycine residue G2 in XopE1, XopE2, and XopJ resulted in cytoplasmic localization of the mutant proteins. Results of hydroxylamine treatments of XopE2 protein extracts suggest that the proteins are additionally anchored in the host cell plasma membrane by palmitoylation. The membrane localization of the effectors strongly influences the phenotypes they trigger in the plant. Agrobacterium-mediated expression of xopE1 and xopJ in Nicotiana benthamiana led to cell-death reactions that, for xopJ, were dependent on the N myristoylation motif. In the case of xopE1(G2A), cell death was more pronounced with the mutant than with the wild-type protein. In addition, XopE2 has an avirulence activity in Solanum pseudocapsicum. PMID- 17918628 TI - Silencing of the N family of resistance genes in Nicotiana edwardsonii compromises the hypersensitive response to tombusviruses. AB - The nontarget effects associated with silencing of the N gene in Nicotiana edwardsonii, an amphidiploid species derived from N. glutinosa and N. clevelandii, have been characterized in this study. The N protein confers resistance to Tobacco mosaic virus (TMV), and is representative of a family of nucleotide-binding site leucine-rich repeat proteins present in N. glutinosa. Previous studies have shown that silencing of the N gene or of other plant genes associated with N-mediated defenses abolishes host resistance to TMV, and this effect can be measured through enhancements in movement or replication of TMV in the N-silenced plants. However, the nontarget effects of gene silencing have not been investigated thoroughly. Notably, are the functions of other resistance (R) genes also affected in experiments designed to silence the N gene? To investigate whether heterologous sequences could silence the N gene, we selected an R gene homolog from N. glutinosa that differed from the N gene by approximately 17%, created a hairpin transgene, and developed transgenic N. edwardsonii plants. Expression of this hairpin in the transgenic N. edwardsonii plants compromised the hypersensitive response to TMV, demonstrating that a single hairpin transgene could silence a block of R genes related by sequence similarity. We then investigated whether the response of N-silenced plants to other viruses would be altered, and found that the hypersensitive response triggered against the tombusviruses Tomato bushy stunt virus and Cymbidium ringspot virus also was compromised. This study indicates that a Tombusvirus R gene shares some homology with the N gene, which could facilitate the cloning of this gene. PMID- 17918629 TI - Allelic variation in the effector genes of the tomato pathogen Cladosporium fulvum reveals different modes of adaptive evolution. AB - The allelic variation in four avirulence (Avr) and four extracellular protein (Ecp)-encoding genes of the tomato pathogen Cladosporium fulvum was analyzed for a worldwide collection of strains. The majority of polymorphisms observed in the Avr genes are deletions, point mutations, or insertions of transposon-like elements that are associated with transitions from avirulence to virulence, indicating adaptive evolution of the Avr genes to the cognate C. fulvum resistance genes that are deployed in commercial tomato lines. Large differences in types of polymorphisms between the Avr genes were observed, especially between Avr2 (indels) and Avr4 (amino-acid substitutions), indicating that selection pressure favors different types of adaptation. In contrast, only a limited number of polymorphisms were observed in the Ecp genes, which mostly involved synonymous modifications. A haplotype network based on the polymorphisms observed in the effector genes revealed a complex pattern of evolution marked by reticulations that suggests the occurrence of genetic recombination in this presumed asexual fungus. This, as well as the identification of strains with identical polymorphisms in Avr and Ecp genes but with opposite mating-type genes, suggests that development of complex races can be the combined result of positive selection and genetic recombination. PMID- 17918630 TI - The role of luxS in the fire blight pathogen Erwinia amylovora is limited to metabolism and does not involve quorum sensing. AB - Erwinia amylovora is a gram-negative phytopathogen that causes fire blight of pome fruit and related members of the family Rosaceae. We sequenced the putative autoinducer-2 (AI-2) synthase gene luxS from E. amylovora. Diversity analysis indicated that this gene is extremely conserved among E. amylovora strains. Quorum sensing mediated by LuxS has been implicated in coordinated gene expression, growth, and virulence in other enterobacteria; however, our evidence suggests this is not the function in E. amylovora. Mutational analysis pointed to a role in colonization of apple blossoms, the primary infection court for fire blight, although little if any role in virulence on apple shoots and pear fruit was observed. Expression of key virulence genes hrpL and dspA/E was reduced in mutants of two E. amylovora strains. Stronger effects on gene expression were observed for metabolic genes involved in the activated methyl cycle with mutants having greater levels of expression. No quorum-sensing effect was observed in coculture experiments with wild-type and mutant strains either in vitro or in apple blossoms. Known receptors essential for AI-2 quorum sensing, the LuxPQ sensor kinase or the Lsr ABC-transporter, are absent in E. amylovora, further suggesting a primarily metabolic role for luxS in this bacterium. PMID- 17918631 TI - An oligonucleotide microarray resource for transcriptional profiling of Bradyrhizobium japonicum. AB - A DNA microarray, comprising 70-mer oligonucleotides, representing 8,453 open reading frames (ORFs), was constructed based on the Bradyrhizobium japonicum strain USDA110 genomic sequence. New annotation predicted 199 additional genes, which were added to the microarray and were shown to be transcribed. These arrays were used to profile transcription in cells under a variety of conditions, including growth in minimal versus rich medium, osmotic stress, and free-living cells versus bacteroids. Increased expression was seen for genes involved in translation, motility, and cell envelope synthesis in rich medium whereas expression increased in minimal medium for genes involved in vitamin biosynthesis and stress responses. Treatment with 50 mM NaCl activated stress-inducible genes but repressed genes involved in chemotaxis and motility. Strikingly, no known transport systems for accumulation of compatible solutes or osmoprotectants were induced in response to osmotic stress. A number of nif, fix, and hup genes, but not all, were upregulated in bacteroids. The B. japonicum type III secretion system, known to be important in early nodulation, was downregulated in bacteroids. The availability of a reliable, low-cost B. japonicum microarray provides a useful tool for functional genomic studies of one of the most agriculturally important bacteria. PMID- 17918632 TI - Detached and attached Arabidopsis leaf assays reveal distinctive defense responses against hemibiotrophic Colletotrichum spp. AB - The agriculturally important genus Colletotrichum is an emerging model pathogen for studying defense in Arabidopsis. During the process of screening for novel pathogenic Colletotrichum isolates on Arabidopsis, we found significant differences in defense responses between detached and attached leaf assays. A near-adapted isolate Colletotrichum linicola A1 could launch a typical infection only on detached, but not attached, Arabidopsis leaves. Remarkably, resistance gene-like locus RCH1-mediated resistance in intact plants also was compromised in detached leaves during the attacks with the virulent reference isolate C. higginsianum. The differences in symptom development between the detached leaf and intact plant assays were further confirmed on defense-defective mutants following inoculation with C. higginsianum, where the greatest inconsistency occurred on ethylene-insensitive mutants. In intact Arabidopsis plants, both the salicylic acid- and ethylene-dependent pathways were required for resistance to C. higginsianum and were associated with induced expression of pathogenesis related genes PR1 and PDF1.2. In contrast, disease symptom development in detached leaves appeared to be uncoupled from these defense pathways and more closely associated with senescence: an observation substantiated by coordinated gene expression analysis and disease symptom development, and chemically and genetically mimicking senescence. PMID- 17918633 TI - [Acute massive pulmonary embolism]. AB - To establish the diagnosis of the acute massive pulmonary embolism is critically urgent for the doctors, because the life saving causal therapy should be introduced as soon as possible. The combined use of the estimated clinical probability and the results of one or more noninvasive examinations increase the accuracy in confirming the diagnosis of embolism. The most important methods to confirm or rule out the suspicion of pulmonary embolism are the noninvasive perfusion pulmonary isotopic scanning, spiral CT, echocardiography and the invasive "gold standard" pulmonary angiography. The life saving causal therapy means the urgent recanalization of the obstructed pulmonary arteries. The surgical embolectomy is a high risk intervention, but the invasive radiological methods are also capable of destroying the emboli mechanically and very effectively. The different thrombolytic treatment methods mean causal therapy too, and these protocols can be used in every hospital. There is no alternative treatment in patients with sudden clinical death caused by acute pulmonary embolism: the drug administration during reanimation should be completed with thrombolysis. PMID- 17918634 TI - [Her2/neu receptor: one of the basic factors rendering tailored therapy feasible in breast cancer patients]. AB - In a considerable part of breast carcinomas the amount of cell surface Her2/neu receptors has been increased such that it becomes the main factor defining the behaviour of the tumor. At the same time, in such amount, this receptor is easily detectable by simple, routine pathological methods. If we are aware of the exact biological characteristics of a given tumor, targeted therapy may be used since an antibody against the Her2 receptor is already available. This drug, called trastuzumab (Herceptin), has been used in advanced breast carcinoma as adjuvant or primary treatment. In clinical trials its advantage in early stage breast carcinomas has been investigated so far. PMID- 17918635 TI - [Endothelial dysfunction and early atherosclerosis in non-differentiated collagenosis]. AB - INTRODUCTION: The most common systemic autoimmune diseases, as the rheumatoid arthritis (RA) and the systemic lupus erythematosus (SLE) are associated with accelerated atherosclerosis and increased cardiovascular morbidity and mortality. The authors' aim was to determine the endothelial dysfunction and the accelerated atherosclerosis in patients with undifferentiated connective tissue disease (UCTD), in a preliminary phase of defined connective tissue diseases. PATIENTS AND METHODS: Twenty-two UCTD patients and twenty age and sex-matched controls were included. Using high-resolution B-mode ultrasound, the authors' measured the intima-media thickness (IMT) of the common carotid artery (CCA), and the diameter of brachial artery at rest, during reactive hyperemia, and after sublingual glyceryl trinitrate administration. The clinical, the demographical status and the serological profile of UCTD patients were also assessed. RESULTS: There was no significant difference between the groups considered to the traditional risk factors. The endothelium dependent vasodilatation was significantly impaired in UCTD patients as compared to the controls (5.3 +/- 3.03% vs. 8.85 +/- 4.02%, P < 0.002). The authors' have not found significant difference between the two groups either at the endothelium independent vasodilatation or at the CCA-IMT. CCA-IMT correlated significantly with the age (R = 0.819, p < 0.001) and with the anti DNA antibody levels (R = 0.563, P < 0.008). CONCLUSIONS: The endothelium dependent vasodilatation of patients with UCTD is reduced before development of a defined connective tissue disease and the potential anti-atherogenic treatment. The endothelium dependent vasodilatation is a more sensitive method to determine an early atherosclerotic process. The authors' found moderate correlation between the CCA-IMT and the anti-DNA antibody levels. PMID- 17918637 TI - [Adenomatoid malformation as peripheral round shadow in lung]. AB - The authors present a case report of a 17 years old female patient who had a peripheral round shadow on the chest X-ray. After the operation the histological examination verified cystic adenomatoid malformation. The cystic adenomatoid malformation is an abnormal growth of the terminal bronchiolar structures. It is a rare disease, but often associates with other developmental abnormalities. It is important in the differential diagnosis to separate this disease from other cystic pulmonary lesions or sequestration. It is usually recognized in antenatal period or early childhood, but rarely in adulthood. It is possible that there is a connection between the adenoid malformation and lung cancer. PMID- 17918638 TI - On the record about EMRs. Advancing technology will save time, money and lives. PMID- 17918636 TI - [Increased IgE-type antibody response to food allergens in irritable bowel syndrome and inflammatory bowel diseases]. AB - BACKGROUND: The irritable bowel syndrome (IBS) is a functional bowel disorder characterized by symptoms of abdominal pain that is associated with disturbed defecation. Crohn's disease (CD) and Ulcerative colitis (UC) are collectively referred to as inflammatory bowel diseases (IBD). IBD appears to result from dysregulated immune response with contributions from genetic predisposition and environmental factors. Among environmental factors the enteric microflora and the components of food (eg. antigens) may play important role in the pathogenesis of IBD. The aim of the authors' study was to detect IgE type antibodies against the most frequent food allergens in patients with IBD and IBS. METHODS: Antibodies against food allergens (IgE type) with ELISA method in patients with CD, UC and IBS were quantitatively measured. The figures were compared to the result of a healthy control group contains the same number of participants, and the frequency of food allergy in these patient groups were determined. RESULTS: On the basis of the study the presence of increased IgE type immune response against any food allergens was 34.5% (p = 0.01) in the group of IBS. In patients with CD or UC the immune response against food allergens was also more frequent than in the control group. The order of frequency of food allergens in IBS was the following: milk protein, soybean, tomato, peanut, egg white. CONCLUSION: As the occurrence of food allergens was significantly higher in IBS group than in the control group, the role of food allergy in the symptoms of IBS can be come up. The frequency of food allergy in patients with diagnosis of IBS, and the similarity of symptoms of this two diseases give an obvious opportunity for the patients with IBS to use a diet avoiding the most frequent food allergens, or take e.g. Na-cromoglycate which is useful therapy in food allergy. PMID- 17918639 TI - Managing liability exposure and safety. Taking risky business out of the MRI suite. PMID- 17918640 TI - Dispensing of excess scrub expenses. PMID- 17918641 TI - A pandemic plan for success. PMID- 17918642 TI - Integrated systems uncover mission-critical information. PMID- 17918643 TI - System ensures fluid specimen collection. PMID- 17918644 TI - Monitoring expenses in an upbeat cardiac market. PMID- 17918645 TI - Hold vendors accountable for proper sterlizing instructions. PMID- 17918646 TI - Is tenure justified? An experimental study of faculty beliefs about tenure, promotion, and academic freedom. AB - The behavioral sciences have come under attack for writings and speech that affront sensitivities. At such times, academic freedom and tenure are invoked to forestall efforts to censure and terminate jobs. We review the history and controversy surrounding academic freedom and tenure, and explore their meaning across different fields, at different institutions, and at different ranks. In a multifactoral experimental survey, 1,004 randomly selected faculty members from top-ranked institutions were asked how colleagues would typically respond when confronted with dilemmas concerning teaching, research, and wrong-doing. Full professors were perceived as being more likely to insist on having the academic freedom to teach unpopular courses, research controversial topics, and whistle blow wrong-doing than were lower-ranked professors (even associate professors with tenure). Everyone thought that others were more likely to exercise academic freedom than they themselves were, and that promotion to full professor was a better predictor of who would exercise academic freedom than was the awarding of tenure. Few differences emerged related either to gender or type of institution, and behavioral scientists' beliefs were similar to scholars from other fields. In addition, no support was found for glib celebrations of tenure's sanctification of broadly defined academic freedoms. These findings challenge the assumption that tenure can be justified on the basis of fostering academic freedom, suggesting the need for a re-examination of the philosophical foundation and practical implications of tenure in today's academy. PMID- 17918647 TI - Why ritualized behavior? Precaution Systems and action parsing in developmental, pathological and cultural rituals. AB - Ritualized behavior, intuitively recognizable by its stereotypy, rigidity, repetition, and apparent lack of rational motivation, is found in a variety of life conditions, customs, and everyday practices: in cultural rituals, whether religious or non-religious; in many children's complicated routines; in the pathology of obsessive-compulsive disorders (OCD); in normal adults around certain stages of the life-cycle, birthing in particular. Combining evidence from evolutionary anthropology, neuropsychology and neuroimaging, we propose an explanation of ritualized behavior in terms of an evolved Precaution System geared to the detection of and reaction to inferred threats to fitness. This system, distinct from fear-systems geared to respond to manifest danger, includes a repertoire of clues for potential danger as well as a repertoire of species typical precautions. In OCD pathology, this system does not supply a negative feedback to the appraisal of potential threats, resulting in doubts about the proper performance of precautions, and repetition of action. Also, anxiety levels focus the attention on low-level gestural units of behavior rather than on the goal-related higher-level units normally used in parsing the action-flow. Normally automatized actions are submitted to cognitive control. This "swamps" working memory, an effect of which is a temporary relief from intrusions but also their long-term strengthening. Normal activation of this Precaution System explains intrusions and ritual behaviors in normal adults. Gradual calibration of the system occurs through childhood rituals. Cultural mimicry of this system's normal input makes cultural rituals attention-grabbing and compelling. A number of empirical predictions follow from this synthetic model. PMID- 17918648 TI - Simultaneous removal of ammonium and phosphate by zeolite synthesized from coal fly ash as influenced by acid treatment. AB - Zeolite synthesized from fly ash (ZFA) without modification is not efficient for the purification of NH4+ and phosphate at low concentrations that occur in real effluents, despite the high potential removal capacity. To develop an effective technique to enhance the removal efficiency of ammonium and phosphate at low concentrations, ZFA was modified with acid treatment and the simultaneous removal of ammonium and phosphate in a wide range of concentration was investigated. It was seen that when compared with untreated ZFA, only the treatment by 0.01 mol/L of H2SO4 significantly improved the removal efficiency of ammonium at low initial concentrations. The behavior was well explained by the pH effect. Treatment by more concentrated H2SO4 led to the deterioration of the ZFA structure and a decrease in the cation exchange capacity. Treatment by 0.01 mol/L H2SO4 improved the removal efficiency of phosphate by ZFA at all initial P concentrations, while the treatment by concentrated H2SO4 (> or = 0.9 mol/L) resulted in a limited maximum phosphate immobilization capacity (PIC). It was concluded that through a previous mild acid treatment (e.g. 0.01 mol/L of H2SO4), ZFA can be used in the simultaneous removal of NH4+ and P at low concentrations in simulating real effluent. PMID- 17918649 TI - Adaptation of Nitraria sphaerocarpa to wind-blown sand environments at the edge of a desert oasis. AB - This study addresses the adaptation of Nitraria sphaerocarpa to blown sand at the edge of a desert oasis with regard to the aspects of soil seed banks, seedlings, and population. Horizontally, the total number of seeds per unit area decreased from the shrub canopy center to intershrub areas, and most seeds were found under shrub canopies. Vertically, the highest proportion of seeds was found at depths of 5-10 cm. The emergence percentage, seedling mass, and seedling height, which were significantly affected by both burial depth and seed size, were highest at the optimal burial depth of 2 cm, and decreased with increasing burial depth in each seed size-class. Although seedling mass was usually greatest for large seeds and least for small seeds at each burial depth, little difference was observed in seedling height at shallow burial depths of 0-3 cm. The population shows a patchy and discontinuous distribution pattern. Population height increases with increasing sand depth. Also the density increases with increasing depth of sand in the desert; however, there is a steady decrease when the depth of sand is more than 100 cm. This result indicates that the depth of sand that is most suitable for the growth of Nitraria sphaerocarpa is 100 cm. The size of the population is significantly correlated with the sand depth, which increases with increasing depth in the desert. PMID- 17918650 TI - [Picture of the month: Ixodes ricinus]. PMID- 17918651 TI - [The intolerable arrogance of the system or how some drug contribution acts]. PMID- 17918652 TI - ["Genuine" neurotics]. PMID- 17918653 TI - [The prescription server requires cooperation]. PMID- 17918654 TI - Outbreak news. Ebola virus haemorrhagic fever, Democratic Republic of the Congo -update. PMID- 17918655 TI - Progress in measles control: Nepal, 2000-2006. PMID- 17918656 TI - Recommended composition of influenza virus vaccines for use in the 2008 influenza season. PMID- 17918657 TI - Medicare and state health care programs: fraud and abuse; safe harbor for federally qualified health centers arrangements under the anti-kickback statute. Final rule. AB - In accordance with section 431 of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA), this final rule sets forth a safe harbor under the anti-kickback statute to protect certain arrangements involving goods, items, services, donations, and loans provided by individuals and entities to certain health centers funded under section 330 of the Public Health Service Act. The goods, items, services, donations, or loans must contribute to the health center's ability to maintain or increase the availability, or enhance the quality, of services available to a medically underserved population. PMID- 17918658 TI - [Should the National Board of Health's treatment recommendations be based on so called "historical analyses"?]. PMID- 17918659 TI - [Depression and ischemic heart disease]. PMID- 17918660 TI - ["Genuine neurotics"--one more time]. PMID- 17918661 TI - UK national audit of sexual health care for people with HIV infection: case-notes audit. AB - The case-notes of 3210 patients with HIV infection were audited. A sexual history was documented within four weeks before or after initial HIV diagnosis in 69% of cases (regional range 45-84%), and in the six months before attendance during the audit interval in 34% (12-53%). An offer of tests for sexually transmitted infections was documented within four weeks before or after HIV diagnosis in 58% (30-83%), and in the prior six months in 28% (14-47%). Syphilis serology was offered in the previous three months to 45% (14-100%) of cases resident in syphilis outbreak areas and to 25% (7-62%) of other cases. Hepatitis B testing was performed for 98% (95-100%) of cases and for hepatitis C, for 91% (79-100%). Cervical cytology results in the past year were documented for 73% (43-94%) of eligible women. Considerable inter-regional variation in performance exists. Interventions are needed to improve the sexual health care of people with HIV infection. PMID- 17918662 TI - HIV-related mortality among tuberculosis patients in The Netherlands, 1993-2001. AB - Human immunodeficiency virus (HIV) infection in tuberculosis (TB) patients in The Netherlands during the period 1993-2001 was associated with an increased risk of death (adjusted odds ratio 4.71, P < 0.002). Age and sex-standardised mortality rates among HIV-infected TB patients decreased significantly over time, from 22.9% in 1993-1995 to 11.8% in 1999-2001 (P < 0.001). No such change was observed for HIV-negative patients. The decrease in mortality is likely due to the more widespread use of highly active antiretroviral therapy (HAART). PMID- 17918663 TI - Abnormal motor movements associated with combining psychostimulants and atypical antipsychotics in children. AB - A random sampling of patients taking a psychostimulant, an atypical antipsychotic, a combination of both, or neither were administered a brief examination to assess the possibility of abnormal motor movements induced by combining these classes of medications. Children on the combination demonstrated significant abnormalities when compared to those on either medication alone. This suggests a need for further study of neurologic risks associated with combining these medications. PMID- 17918664 TI - Allelic imbalance of HER-2 codon 655 polymorphism among different religious/ethnic populations of northern Greece and its association with the development and the malignant phenotype of breast cancer. AB - Alterations of c-erbB-2 (neu or HER-2) proto-oncogene have been associated with carcinogenesis and poor prognosis of breast cancer. A single nucleotide polymorphism (SNP) at codon 655 resulting in a G to A transition (Ile655Val) in the transmembrane domain-coding region of this gene has been associated with an increased risk of breast cancer. This study aims to determine the prevalence of the HER-2 genotype and its association with breast cancer in the Greek Christian and Greek Muslim population of Thrace, Greece. In this case-control study, we genotyped 56 patients (43 Christians and 13 Muslims) with primary breast cancer and 45 healthy women (32 Christians and 13 Muslims) for the Ile655Val polymorphism, with the PCR-RFLP method. The Val allele and the Val-containing genotypes were significantly more frequent in Muslims than in Christians (p=0.020 and p=0.008, respectively). Among the Greek Christian population, a 5-fold and a 3.1-fold increase in risk of breast cancer was associated with the Val-Val genotype and the Ile-Val or Val-Val genotypes (95% CI, 1.3-18.4; p=0.017 and aOR, 3.1; 95% CI, 1.2-8.3; p=0.025; respectively) compared to homozygous Ile-Ile. No significant association was found in the Muslim population. Among the entire cohort, the Val allele confers a modest increase in breast cancer risk (OR, 2.6; 95% CI, 0.9-7.6; p=0.076, for Val-Val and OR, 2.2; 95% CI, 0.9-5.2; p=0.079 for Ile-Val or Val-Val). This effect was even more pronounced in younger women. Among breast cancer patients, invasive carcinomas, low differentiation tumors, advanced stages, positive lymph nodes, high number of lymph nodes and HER-2 overexpression were more frequent in patients with allele Val than those with allele Ile. Our study proposes the allelic imbalance of Ile655Val polymorphism between Greek Christian and Greek Muslim populations of Thrace contributes to the inconsistent association between this SNP and breast cancer risk across these two different ethnic groups. The association of the HER-2 genotype with clinicopathologic characteristics and HER-2 expression may indicate its possible implication on the more aggressive phenotype. PMID- 17918665 TI - Clinical, pathological and molecular characteristics of newly diagnosed breast cancers. AB - Selection of breast carcinoma therapy is based on standard prognostic markers, such as tumor size, infiltration of regional lymph nodes, tumor grade, and expression of hormonal receptors. Insufficient treatment results stimulate a search for new markers which may lead to a more precise characterization of these tumors and to a more effective treatment. In our study we determined essential clinical and histopathological characteristics of non-metastasizing breast cancer - primary tumor size, involvement of the regional lymph nodes, expression of hormonal receptors and a status of ERBB-2 protein (HER-2), DNA ploidy, and their possible inter-correlation. In this study 77 patients were analyzed. The mean age was 59.3 years. Tumor stage T1 was found in 53%, T2 in 39%, T3 in 5% of patients. 57% of patients did not show any metastases in the axillary lymph nodes. A higher tumor grade 3 was seen mainly in larger tumors, in 62% of T2 and 66% of T3 tumors; 77% of carcinomas expressed hormonal receptors. HER-2 expression was shown in 21 T1 tumors, 13 T2 tumors, and 1 T3 tumor. 47 tumors were diploid. 13 T1 tumors, 14 T2 tumors, and 2 T3 tumors were aneuploid. Any significant correlation among staging T, N and ERBB-2 expression, hormonal receptors expression, tumor grade and DNA ploidy was found. PMID- 17918666 TI - Judicial interventions for morbidly obese children. PMID- 17918668 TI - Proving medical necessity in claims for Medicare reimbursement: Eleventh Circuit finds a certificate of medical necessity does not unequivocally establish medical necessity under Part B of the Medicare Act--Gulfcoast Medical Supply, Inc. v. Secretary, Department of Health and Human Services. PMID- 17918667 TI - Tort law: Desiano and the Second Circuit's repudiation of de jure tort reform. PMID- 17918669 TI - Insurance: New York rejects religious challenge to law requiring employers to provide insurance coverage for contraceptives--Catholic Charities of the Diocese of Albany v. Serio. PMID- 17918670 TI - One year outcome after surgery for lumbar disc herniation: a comparison of reoperated and not reoperated patients. AB - OBJECTIVES: To assess the disability level and quality of life of the patients who had been reoperated and not reoperated because of lumbar disc herniation. METHODS: 34 reoperated and 34 not reoperated patients with lumbar disc herniation were retrospectively included in the study. The disability levels of the patients were evaluated with the Oswestry Disability Index and the the quality of life was evaluated with SF-36. The measurements were repeated in the 2nd, 6th and 12th months after the operation. RESULTS: The disability measurements of the reoperated and not reoperated patients were found to be significantly different in the 2nd, 6th and 12th months (p < 0.05); and it was noticed that the reoperated group had more disability levels than not reoperated patients in all the assessments. SF-36 measurement in the 2nd month showed significant differences in all parameters except pain (p < 0.05). There were important differences in all parameters of quality of life in the 6th month and significant differences were found in all parameters of quality of life except physical and emotional role limitation in the 12th month (p < 0.05) in favour of the not reoperated group. CONCLUSIONS: The increase in the number of operations affects the disability level and quality of life negatively. PMID- 17918672 TI - Intracarotid L-arginine reverses motor evoked potential changes in experimental cerebral vasospasm. AB - OBJECTIVE: To investigate motor evoked potentials during short term L-arginine infusion in cerebral vasospasm after experimental subarachnoid hemorrhage. METHODS: Three groups were designated for this study: control group, subarachnoid hemorrhage-saline infusion group, and subarachnoid hemorrhage-L-arginine infusion group. A subarachnoid hemorrhage was created by intracisternal injection of autologous blood in New Zealand rabbits. At the fourth day of subarachnoid hemorrhage, latency and amplitude of motor evoked potentials were recorded during intracarotid saline and L-arginine infusion, and compared with motor evoked potential parameters of the control group. RESULTS: Motor evoked potential latencies were increased, and amplitudes were decreased in all animals before saline and L-arginine infusion on the fourth day of subarachnoid hemorrhage. A decrease in latencies as well as an increase in motor evoked potential amplitudes was observed with short-term intracarotid L-arginine infusion. CONCLUSION: Intracarotid short term L-arginine infusion significantly improves motor evoked potential parameters after experimental subarachnoid hemorrhage. PMID- 17918671 TI - Late decompression in patients with acute facial nerve paralysis after temporal bone fracture. AB - OBJECTIVE: The aim of this paper was to address the management of acute facial nerve paralysis after temporal bone fracture and the outcomes of late decompression. METHODS: The study design was a retrospective review of eight patients who underwent late decompression of acute facial nerve paralysis due to temporal bone fracture involving the geniculate region. Pre-operative electrophysiological testing showed total degeneration of the facial nerve in all patients. The mean operation period was 70.1 +/- 54.8 days after the trauma. A pure middle fossa approach and combined approach included middle cranial fossa and transmastoid approaches and were used in six and two patients, respectively. Evaluation of the facial nerve function was graded according to the House Brackmann grading scale. RESULTS: The mean follow up period was 5.7 +/- 3.2 years, ranging from 3 months to 10 years. Six patients showed 3 score of House Brackmann recovery of facial nerve function in long-term follow-up and the last 2 patients still need time for their final House-Brackmann evaluation. CONCLUSIONS: Surgery should be performed if serial electroneurography and electromyography demonstrate more than 90% degeneration and total denervation potentials, respectively, of nerve fibers. Based on the outcomes observed, late facial nerve decompression may have still beneficial effects in patients who could not be operated on early. PMID- 17918673 TI - Subcutaneous granuloma annulare of the scalp in childhood: a case report and review of the literature. AB - Granuloma annulare is a benign inflammatory skin lesion of unknown etiology that is usually seen in adults and children and subtypes of it includes localized granuloma annulare, generalized granuloma annulare, subcutaneous granuloma annulare and arcuate dermal erythema. Etiology and pathogenesis of granuloma annulare are obscure, although there is much evidence for an immunologic mechanism. Precipitating factors are insect bites, sunburn, photochemotherapy, drugs, physical trauma, acute phlebitis and sepsis after surgery. Some investigators were suggested a relationship of granuloma annulare to a latent or clinically manifest diabetes or rheumatoid arthritis. In contrast, an association of subcutaneous granuloma annulare with these diseases in childhood has not been reported in the literature. Subcutaneous granuloma annulare of the scalp is rare lesion in childhood and nodules on the scalp are usually non-, or slightly mobile, whereas lesions on the extremities are freely mobile. For definitive diagnosis, a biopsy should be performed but wide surgical intervention or medical treatment is not indicated. In case of recurrence, no additional diagnostic studies are necessary. PMID- 17918674 TI - Huge cavernoma with massive intracerebral hemorrhage in a child. AB - Cavernous malformations are benign vascular lesions of the central nervous system that lack intervening normal brain parenchyma. They can be seen almost anywhere that normal vasculature is available. Lesions are raspberry-like, thin-walled vascular sinusoids without smooth muscles containing hemosiderin deposits. Cerebral cavernous malformations are characterized by small bleedings. Their size varies from a few millimeters to 2- 3 centimeters. Giant cases are rare. Also referred to as cavernoma, these lesions rarely lead to intracerebral hematomas that threaten life. In this report, we have presented a 14-year-old patient with a giant cavernoma leading to a life-threatening massive intracerebral hematoma. PMID- 17918675 TI - Craniopagus, Laleh and Ladan twins, sagital sinus. AB - Adult craniopagus twins aged 29 years who succumbed to the surgical separation are being presented posthumously. Neuroimaging studies had revealed a total type 3 craniopagus with a common superior sagittal sinus beginning at its distal half and draining into a common Torcular Herophili. In July 2003, neurosurgeons at Singapore embarked on the mission to tackle the separation of the most complex and challenging adult craniopagus of the present millennium, unfortunately ending up with devastating results. The authors intend to expand upon their life style, shared intracranial structures, the potential risks of a one-stage procedure as carried out in this case and finally offer some suggestions in tackling such rare and complex cases in neurosurgery. PMID- 17918676 TI - Cavernous hemangioma of temporalis muscle: report of a case and review of the literature. AB - Hemangiomas are tumors of vascular origin comprising approximately 7% of all benign tumors. Intramuscular hemangioma is a rare condition and hemangiomas of the head and neck make up less than 15% of intramuscular hemangiomas. Temporalis muscle is an uncommon location for intramuscular hemangioma and seldom reported in the literature. Radiological methods are generally insufficient for the correct diagnosis and surgery is the treatment of choice to exclude malignancy and for adequate treatment of these lesions. A 37-year-old male was admitted with a slowly growing painless mass in his right temporal fossa. The lesion was surgically excised and histopathology confirmed the diagnosis of cavernous hemangioma. Diagnosis and treatment modalities for temporalis muscle hemangiomas are discussed. PMID- 17918677 TI - Enlargement of postoperative aqueductal air due to elevated body temperature. Case report. AB - Pneumocephalus has been reported after posterior fossa surgery especially with procedures performed in the sitting position. The gravitational effect is the decisive factor in the development of pneumocephalus. The entrapped air in the aqueduct may enlarge due to several factors such as elevated body temperature and may cause to deterioration in neurological status. We report a rare case of tension pneumocephalus associated with the enlargement of massive air in aqueduct due to elevated body temperature, following removal of a cervicomedullary tumor. We believe her neurological deterioration was due to the compression of the reticular formation by dilatation of postoperative air in the aqueduct due to the elevation of her body temperature. PMID- 17918678 TI - Treatment differences in cases with venous angioma. AB - Venous angiomas were found to be the most common cerebral vascular malformations, composing 63% of such lesions in two autopsy series. Annual bleeding risk associated with venous angiomas is about 0.22 % per year. Venous angiomas are generally silent lesions because of their dynamic features, and are low flow and low pressure vascular structures draining normal brain tissue. An angioma rarely causes symptoms such as bleeding, seizure, hemifacial spasm, trigeminal neuralgia, aqueduct compression, nonhemorrhagic infarction and thrombosis of the draining vein. Even if it should bleed, the lesion can be managed conservatively in asymptomatic or mildly symptomatic patients. In this paper we report two venous angioma cases. The first patient bled twice in a short period of time and the angioma was located at the posterior fossa next to the left lateral recess. The second patient recently suffered a cerebral stroke that was located in the vicinity of the right caudate nucleus and not associated with the venous angioma that was located next to the left caudate nucleus. This patient had been under warfarin sodium treatment for 14 years due to his previous coronary artery bypass surgery, but unknowingly there was a venous angioma located next to the caudate nucleus. PMID- 17918679 TI - Unrecognized peripheral nerve lesions in a traumatic brain injury patient. AB - Patients with traumatic brain injury constitute a diagnostic challenge since peripheral nerve injuries may be overlooked due to cognitive dysfunction and priority given to life-sustaining measures. Electromyography may be helpful in the differential diagnosis of weakness and atrophy. Problems specific for the traumatic brain injury patients, namely heterotopic ossification, hypertrophic callus formation and myositis ossificans should be considered by the physician. We report a 15-year-old patient involved in a pedestrian motor vehicle accident with traumatic brain injury. He had weakness and atrophy of the left upper extremity. Electromyographic examination revealed axillary nerve injury and carpal tunnel syndrome. Differential diagnosis of atrophy and weakness in traumatic brain injury patients is discussed. PMID- 17918680 TI - Ischemic stroke complicated by a brain abscess: a case report and review of the literature. AB - Ischemic stroke and brain abscess are life threatening, severe medical conditions requiring urgent and comprehensive evaluation. Brain abscess is a very rare and dangerous complication of ischemic stroke that can be treated successfully when diagnosed. Association of these two life threatening condition may cause catastrophic results. In this report, we present our experience with an ischemic stroke case complicated by a brain abscess under the light of previous literature. A 64-year-old patient with a cerebral infarction complicated by a cerebral abscess is reported with its clinical and radiological features. Because of their insidious and opportunistic nature, brain abscess complicating cerebral infarcts are pathologies requiring a high index of suspicion for proper treatment and favorable results. PMID- 17918681 TI - Removal of clival chordoma in an adolescent thorough combned pterional transsylvian and anterior temporal approach. AB - Extensive and aggressive surgical removal is treatment of choice for patients who have chordomas of the cranial base. Well-developed microsurgical techniques, as well as good surgical judgment learned from experience are essential to avoid major morbidity. A 14-year-old female presented with progressive headaches and diplopia for three months. Cranial magnetic resonance imaging demonstrated a well circumscribed mass in the clival region. The mass was totally excised via right combined pterional transsylvian and anterior temporal approach (+ orbitozygomatic osteotomy). The tumor was located extradurally. The resected tumor had the typical histological and immunohistochemical characteristics of chordoma. No radiation therapy or chemotherapy was administered. PMID- 17918682 TI - Bypass to the intracranial giant or large internal carotid artery aneurysms: superficial temporal artery to middle cerebral artery bypass re-visited. AB - OBJECTIVE: Hunterian ligation of the internal carotid artery is an acceptable treatment modality for inoperable intracranial carotid aneurysms. Despite the risk of thrombo-embolic complications, ligation together with superficial temporal artery-middle cerebral artery (STA-MCA) bypass surgery is vital in some patients. Our aim is to present our experience in five patients in whom STA-MCA bypass and balloon occlusion were performed due to inoperable intracranial carotid artery aneurysms. METHODS: During the last five years, five patients with inoperable internal carotid artery (ICA) aneurysms were treated by STA-MCA bypass surgery followed by permanent endovascular ICA occlusion. Patients were selected for bypass surgery on the basis of failing balloon test occlusion. Patients were managed in the intensive care unit after surgical and endovascular procedures. RESULTS: Clinical improvement was noted in all patients and no major complication during the follow-up was seen. Aneurysmal thrombosis was confirmed in all patients. CONCLUSION: Hunterian ligation associated with bypass surgery is an effective treatment method in selected patients. The following points should be considered for a good outcome: (1) experienced surgeon for bypass surgery, (2) experienced neuroradiologist for endovascular occlusion of the parent vessel as close to the aneurysm neck as possible, and (3) judicious postoperative management by means of anticoagulation, fluid replacement, and pressure control. PMID- 17918683 TI - Ultrasound kidney image analysis for computerized disorder identification and classification using content descriptive power spectral features. AB - The objective of this work is to classify few important kidney categories by characterizing the tissues of kidney region using the unique power spectral features with ultrasound as imaging modality. The images are acquired from male and female subjects of age 45 +/- 15 years. Three kidney categories namely normal, medical renal diseases and cortical cyst are considered for the analysis. The acquired images are initially preprocessed to retain the pixels-of-interest. The proposed features depend on the spatial distribution of spectral components in the kidney region. A set of power spectral features P(T)(W1), P(T)(W2), P(T W12)R1, P(T-W12)R2, P(T-W1d)R3 and P(T-W1d)R4 are estimated at the specific cut off frequencies omega(rc1), and omega(rc2) in the spectrum and by considering global mean total power. The results obtained show that the features are highly content descriptive and provide discrete range of values for each kidney category. Such isolated feature values facilitate to identify the kidney categories objectively which may be used as a secondary observer. The proposed method and features also explores the possibility of implementing computer-aided diagnosis system exclusively for US kidney images. PMID- 17918684 TI - Is the availability of hospital IT applications associated with a hospital's risk adjusted incidence rate for patient safety indicators: results from 66 Georgia hospitals. AB - This study examines the associations between the availability of IT applications in a hospital and that hospital's risk adjusted incidence rate per 1,000 hospitalizations for Agency for Healthcare Research and Quality's (AHRQ) 15 Patient Safety Indicators (PSIs). The study population consists of a convenience sample of 66 community hospitals in Georgia that completed a Hospital IT survey by December 2003 and provided data to Georgia Hospital Discharge Data Set during 2004. AHRQ's PSI software was used to estimate risk adjusted incidence rates. Differences in means, Pearson correlation coefficients, and multivariate regression analysis were used to determine if the availability of IT applications were associated with better PSI outcomes. This study finds very little statistically significant correlation between the availability of IT applications and risk adjusted PSI incident rate per 1,000 PMID- 17918685 TI - A classification system for stenosis from mitral valve Doppler signals using adaptive network based fuzzy inference system. AB - In this study, cardiac Doppler parameters were studied in 60 patients with mitral valve stenosis and compared with 41 ages and sex matched healthy controls. Firstly, the sonograms which represent the changes in Doppler frequency with respect to time were performed from mitral valve Doppler signals using short time Fourier transformation (STFT) method. Secondly, the envelopes of these sonograms and data set depicted from sonogram envelopes were acquired. Finally, the processed data set are applied to the proposed adaptive network based fuzzy inference system (ANFIS) model has potential in classifying the mitral valve Doppler signals. This result confirms that our technique contribute to the detection of mitral valve stenosis and our method offers more reliable information than looking at the sonogram on the Doppler screen and making a decision from the visual inspection. The proposed ANFIS model combined the neural network with adaptive capabilities and qualitative approach of fuzzy logic. The obtained results show that 98% correct classification was achieved, whereas two false classifications have been observed for the test group of 101 people. PMID- 17918686 TI - EBM metadata based on Dublin Core better presenting validity of clinical trials. AB - To help clinicians find better evidence, a metadata schema for Evidence Based Medicine (EBM) is developed. Dublin Core metadata standard (DC) was adopted to help build a metadata schema. An experimental system was developed to test the validity of the metadata and full text papers of clinical therapy on stomach ulcer extracted using PubMed. An EBM metadata schema was developed. Citations were created from original papers using the metadata schema. Three clinicians evaluated papers by utilizing metadata and full texts respectively. Agreement of evaluation was analyzed, and the result on weighted kappa was 0.55 (95% CI, 0.42 0.67). It reveals that there is moderate agreement between evaluation of metadata citations and full texts. It is possible to use the metadata to select papers before reading the full texts. A further study should be made to prove the applicability of the metadata in the real world setting. PMID- 17918687 TI - The study on the intellectual analysis algorithm for oriental pulse parameters. AB - The digital pulse diagnosis system has been developed on the basis of a systematic pulse diagnosis method. This study has applied the diagnosis algorithm to 60 patients at an oriental medicine hospital. The diagnosis algorithm had been developed on the basis of the pulse wave data acquired through the sensor of the system. The test examined the patients' pulse waves by the two methods of superficially and deeply checking them on the right and left wrists respectively. Since each method checked three parts of each wrist, which are Chon, Kwan, and Chuk, a total of 12 pulse waves were measured per patient. This study presents diagnosis parameters for clinical diagnoses by analyzing structural elements of pulse waveforms. As a result of having comparatively analyzed the result of 720 clinical data diagnosed by the digital pulse diagnosis system, developed by the study and the clinical results of oriental medicine doctors, the identical clinical opinion amounted to as much as 96.25%. The clinical application of the digital pulse diagnosis system developed by this study is of great significance in the sense that it heightened the objectivity and accuracy of the diagnosis. In the future, there should be continued research for establishing the pulse diagnosis database through incessant clinical tests. PMID- 17918688 TI - Quantum noise suppression in X-ray images using fuzzy 2-D Wiener filter. AB - The noisy images are caused by decreasing quantity of the produced X-ray due to the deformation of the X-ray tube's anode. While obtaining the image from low quantity X-ray, shot noise or quantum noise occurs, and this decreases the quality of the image. The aim of this study is to define the novel method called as Fuzzy 2-D Weiner filter (FWF-2D), which suppresses the shot noise from noisy image by avoiding any harm to the image details. With this filtering technique, it is possible to preserve the sharp-edge and the details of the image without any damage during suppressing the noise from images. FWF-2D is one of the best techniques to suppress the corrupted quantum noises due to damaged X-ray tube. The fuzzy rules used in this filter are aimed to distinguish noise pixels from image ones, and the Wiener Filter is working to remove noise pixels distinguished by these fuzzy rules. It is then possible to obtain clean images from damaged X ray tubes by using FWF-2D technique. PMID- 17918689 TI - Comparison of the levels of accuracy of an artificial neural network model and a logistic regression model for the diagnosis of acute appendicitis. AB - An accurate diagnosis of acute appendicitis in the early stage is often difficult, and decision support tools to improve such a diagnosis might be required. This study compared the levels of accuracy of artificial neural network models and logistic regression models for the diagnosis of acute appendicitis. Data from 169 patients presenting with acute abdomen were used for the analyses. Nine variables were used for the evaluation of the accuracy of the two models. The constructed models were validated by the ".632+ bootstrap method". The levels of accuracy of the two models for diagnosis were compared by error rate and areas under receiver operating characteristic curves. The artificial neural network models provided more accurate results than did the logistic regression models for both indices, especially when categorical variables or normalized variables were used. The most accurate diagnosis was obtained by the artificial neural network model using normalized variables. PMID- 17918690 TI - Factors influencing community health centers' efficiency: a latent growth curve modeling approach. AB - The objective of study is to examine factors affecting the variation in technical and cost efficiency of community health centers (CHCs). A panel study design was formulated to examine the relationships among the contextual, organizational structural, and performance variables. Data Envelopment Analysis (DEA) of technical efficiency and latent growth curve modeling of multi-wave technical and cost efficiency were performed. Regardless of the efficiency measures, CHC efficiency was influenced more by contextual factors than organizational structural factors. The study confirms the independent and additive influences of contextual and organizational predictors on efficiency. The change in CHC technical efficiency positively affects the change in CHC cost efficiency. The practical implication of this finding is that healthcare managers can simultaneously optimize both technical and cost efficiency through appropriate use of inputs to generate optimal outputs. An innovative solution is to employ decision support software to prepare an expert system to assist poorly performing CHCs to achieve better cost efficiency through optimizing technical efficiency. PMID- 17918691 TI - Organizational and environmental determinants of hospital EMR adoption: a national study. AB - The recent focus on health care quality improvement and cost containment has led some policymakers and practitioners to advocate the adoption of health information technology. One such technology is the Electronic Medical Record (EMR), which is predicted to change and improve health care in the USA. Little is known about factors that influence hospital adoption of this relatively new technology. The purpose of this paper is to determine the national prevalence of EMR adoption in acute care hospitals while examining the organizational and environmental correlates using a Resource Dependence Theoretical Perspective. Significant predictors of hospital EMR use may indicate barriers to use for some hospitals and can be used to guide policy. This study uses a non-experimental cross sectional design to examine hospital EMR use in 2004. A logistic regression approach is used to determine the correlations between hospital EMR use and organizational and environmental characteristics. Hospital EMR use was identified using the HIMSS Analytics data. Organizational and environmental variables were measured using data from the AHA, CMS (financial and case mix) and ARF. Hospital EMR adoption is significantly associated with environmental uncertainty, type of system affiliation, size, and urbanness. The effects of competition, munificence, ownership, teaching status, public payer mix, and operating margin were not statistically significant. Significant predictors of hospital EMR adoption represent barriers that may prevent certain hospitals from obtaining and using EMRs. These hospitals include those that are smaller, more rural, non-system affiliated, and in areas of low environmental uncertainty. Since EMR adoption may be an organizational survival strategy for hospitals to improve quality and efficiency, hospitals that are at risk of missing the wave of implementation should be offered services and incentives to enable them to implement and maintain EMR systems. PMID- 17918692 TI - Home Internet use among hospice service recipients: recommendations for Web-based interventions. AB - A growing number of researchers are exploring strategies to improve hospice care through the use of web-based technologies. This study of 50 hospice patients and caregivers was conducted in order to obtain data describing home Internet use among hospice service recipients. Over half (58%) of respondents reported having home Internet access, with most using a dial-up connection. Primary reasons for accessing the web included e-mail and information searches. Findings suggest that the hospice industry should explore adopting web-based technologies as a strategy to enhance rather than replace traditional care. Providers must consider the strengths and potential limitations of patients and caregivers when designing online services. Specific recommendations for web-based hospice interventions are discussed at length. PMID- 17918693 TI - Neural network-based diagnosing for optic nerve disease from visual-evoked potential. AB - In this paper, we purpose a diagnostic procedure to identify the optic nerve disease from visual evoked potential (VEP) signals using an Artificial Neural Network (ANN). Multilayer feed forward ANN trained with a Levenberg Marquart backpropagation algorithm was implemented. The correct classification rate was 96.87% for subjects having optic nerve disease and 96.66% for healthy subjects. The end results are classified as healthy and diseased. Testing results were found to be compliant with the expected results that are derived from the physician's direct diagnosis, angiography, VEP and pattern electroretinography. The stated results show that the proposed method could point out the ability of design of a new intelligent assistance diagnosis system. PMID- 17918694 TI - Methods to evaluate health information systems in healthcare settings: a literature review. AB - Although information technology (IT)-based applications in healthcare have existed for more than three decades, methods to evaluate outputs and outcomes of the use of IT-based systems in medical informatics is still a challenge for decision makers, as well as to those who want to measure the effects of ICT in healthcare settings. The aim of this paper is to review published articles in the area evaluations of IT-based systems in order to gain knowledge about methodologies used and findings obtained from the evaluation of IT-based systems applied in healthcare settings. The literature review includes studies of IT based systems between 2003 and 2005. The findings show that economic and organizational aspects dominate evaluation studies in this area. However, the results focus mostly on positive outputs such as user satisfaction, financial benefits and improved organizational work. This review shows that there is no standard framework for evaluation effects and outputs of implementation and use of IT in the healthcare setting and that until today no studies explore the impact of IT on the healthcare system' productivity and effectiveness. PMID- 17918696 TI - Acute stretch perception alteration contributes to the success of the PNF "contract-relax" stretch. AB - CONTEXT: Some researchers have suggested that an alteration of stretch perception could be responsible for the success of the contract-relax (CR) stretch, a stretch technique derived from proprioceptive neuromuscular facilitation (PNF). OBJECTIVE: This study was conducted to determine if the alteration of the stretch perception is a possible explanation for the range of motion (ROM) gains of the CR stretch. PARTICIPANTS: Eighteen subjects performed two stretches in randomized order: the slow stretch and the CR stretch. MAIN OUTCOME MEASURE: The stretch intensity was controlled. The stretch force was measured and compared between the slow stretch and CR stretch. RESULTS: There was a significant difference between the stretch force that could be applied in the PNF stretch (126.0 N) and the slow stretch (108.4 N); P = 0.00086. The average stretch tolerance progressively increased with successive trials from 120.6 N in the first trial to 132.4 N in the fourth trial. CONCLUSION: The alteration of stretch perception plays a role in the success of the CR form of PNF stretching. At least four repetitions of the CR stretch are recommended to get the greatest ROM gain. PMID- 17918695 TI - A comparison of two Thera-Band training rehabilitation protocols on postural control. AB - CONTEXT: Thera-Band (TB) exercises are commonly utilized in ankle rehabilitation, but previous studies have shown inconsistent results. OBJECTIVE: To compare two TB protocols among healthy subjects in improving postural control. DESIGN: Mixed model design. SETTING: Research laboratory. PARTICIPANTS: Thirty healthy subjects divided into a control (CON), three times/week (TB3) or five times/week (TB5) group. INTERVENTION: Training groups completed TB quick-kick protocols for six weeks either three (TB3) or five times (TB5) per week. MAIN OUTCOME MEASURE: Center of pressure velocity in the anterior/posterior (COPVX) and medial/lateral (COPVY) directions. RESULTS: There were no differences related to Group or Side. COPVX in the eyes open (EO) condition increased pre-post. COPVY decreased pre post. CONCLUSION: The lack of differences between Groups and Side indicates these specific TB training protocols did not impact static postural control differently among healthy subjects. PMID- 17918697 TI - Rehabilitation adherence in sport injury: sport physiotherapists' perceptions. AB - CONTEXT: Athlete's adherence behavior can influence the outcome of a rehabilitation program. OBJECTIVE: To draw on sport physiotherapists' expert knowledge to increase understanding of adherence issues in practice and identify factors that influence adherence and strategies that can be used to enhance adherence. DESIGN: An interview design with inductive content analysis. PARTICIPANTS: Nine (6 women and 3 men) experienced sports physiotherapists. RESULTS: Under-adherence and over-adherence were issues in practice for some practitioners, and adherence was viewed as important for successful recovery from injury. Three higher order themes emerged relating to the influence of athlete characteristics, situational characteristics, and characteristics of the injury and rehabilitation on both facilitating and preventing rehabilitation adherence. Strategies to improve adherence in practice emerged from the data and broadly addressed the key determinants of adherence. CONCLUSION: Adherence to rehabilitation is influenced by a number of factors and strategies to enhance adherence are identified. PMID- 17918698 TI - Views of chartered physiotherapists on the psychological content of their practice: a follow-up survey in the UK. AB - OBJECTIVE: To assess, on a national level, the views of chartered physiotherapists with regard to the psychological content of physiotherapy practice. DESIGN: A postal survey to a national list of sport injury and physiotherapy clinics was employed. PARTICIPANTS: A total of 361 responses were included in the descriptive statistical and qualitative analyses. MEASUREMENTS: The Physiotherapist and Sport Psychology Questionnaire (PSPQ). RESULTS: On average, physiotherapists felt that athletes were psychologically affected 83% of the time when injured. Key psychological characteristics were also identified in athletes who cope/do not cope successfully with their injuries. Physiotherapists reported using psychological techniques in their work and expressed the need for further training in the field. Only 24.1% of the physiotherapists stated having accesses to accredited sport psychologists. CONCLUSIONS: Results suggest that UK physiotherapists possess practical experiences and good awareness for psychological aspects of injuries and acknowledge the importance of treating a range of psychological conditions. PMID- 17918699 TI - Ultrasound heating is curvilinear in nature and varies between transducers from the same manufacturer. AB - CONTEXT: Ultrasound heating rates are known to differ between various manufacturers; it is unknown whether this difference exists within a manufacturer. OBJECTIVE: Determine if intramuscular heating differences exist between transducers from the same manufacturer. STUDY DESIGN: 3 x 10 repeated measures. Independent variables were Transducer (A, B, and C) and Time (10-min time points during the treatment). SETTING: Controlled laboratory. PARTICIPANTS: Twelve volunteers (M = 4, F = 8; age: 23 +/- 4 years; calf-girth: 37.94 +/- 4.16 cm; calf-skinfold: 27 +/- 17 mm). INTERVENTION: Three 10-min 1MHz continuous ultrasound treatments performed at an intensity of 1.2 W/cm2, over an area 2x transducer. MAIN OUTCOME MEASURES: Calf temperature increase. RESULTS: Heating curve generated for each transducer were significantly different (P = .034) but the overall temperature increases following 10 minutes of treatment were within 0.1 degree C (F = 1.023 P = .573). CONCLUSION: Heating curves differ between transducers from the same manufacturer but peak heating at 10 minutes was similar. PMID- 17918700 TI - The reliability of the lateral step test. AB - CONTEXT: Functional strength measures correlate more closely with functional performance than non-functional strength measures. OBJECTIVES: To determine the reliability of the lateral step test as a measure of maximal strength. DESIGN: Intertester repeated measures. SETTING: Research laboratory. PARTICIPANTS: Twenty four healthy, pain free subjects. INTERVENTION: Two protocols (A and B) were evaluated. The protocols were identical except protocol B involved a three second pause. Participants performed a one repetition maximum (1RM) for each protocol on two occasions separated by one week. MAIN OUTCOME MEASURES: Step height (nearest cm) representing 1RM. RESULTS: Both protocols demonstrated excellent reliability, protocol A: ICC = 0.94 (95% CI, 0.87 to 0.97), SEM = 1.47 cm. Protocol B: ICC= 0.94 (95% CI, 0.85 to 0.97). Percent close agreement within 2 cm was 83.3% for protocol A and 79.1% for protocol B. CONCLUSION: Both protocols demonstrated excellent inter-tester reliability as measures of functional lower limb strength. PMID- 17918701 TI - Effects of a resistance training program in multiple sclerosis Spanish patients: a pilot study. AB - CONTEXT: Physical exercise is regarded as a useful tool in the treatment of multiple sclerosis (MS). Generally, physical rehabilitation have been based on the prescription of aerobic exercises, while fewer programs have been aimed at developing muscular strength. OBJECTIVE: To establish whether the physical fitness of MS sufferers can be improved by a training program for developing muscular strength. DESIGN: Before and after study SETTING: University multipurpose room PARTICIPANTS: 36 patients, all able to walk, belonging to the Leon Multiple Sclerosis Association. INTERVENTIONS: The physical exercise programme consisted in resistance training sessions, based mainly on callisthenic, or bodyweight, exercises, during six weeks. PMID- 17918702 TI - [Palindromic sequence-mediated de novo chromosome translocation in sperm samples derived from normal and oligospermic males]. AB - OBJECTIVE: To systematically explore the occurrence of a novel type of chromosome translocation in human sperm samples. METHODS: Specific translocation junction fragments were quantified using nested and/or multi-nested PCR in sperm DNA derived from 28 oligospermic patients and 32 normal controls. RESULTS: t(11;22) was detected in 49 samples. At least 4 samples were found to have t(1;22) (p21.2;q11.2), t(17;22) (q11;q11) or t(X;22) (q27;q11). The mutation rate seemed to be associated not with age or semen volume, but with sperm concentration (r = 0.389, P < 0.05) and motility (r = -0.397, P < 0.05). Correlation was not found between homology of palindromic sequences and mutation rate. CONCLUSION: Palindromic sequence mediated chromosome translocation is common in human sperm, and associated with sperm concentration and motility. Measurement of such mutations may provide a molecular-level reference for assessing sperm quality. PMID- 17918703 TI - [Effects of interferon-gamma and tumor necrosis factor-alpha on the fertilizing capacity of human sperm and their mechanisms]. AB - OBJECTIVE: To investigate the effects of interferon-gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-gamma) on the sperm acrosin activity and the rate of acrosome reaction and to probe into their mechanisms. METHODS: Thirty-six nearly normal semen samples were treated with IFN-gamma and/or TNF-alpha after isolated by 75% Percoll. The sperm acrosin activity was tested by the method of BAEE/ADH Unity, the rate of acrosome reaction observed by Triple-stain technique, the NO concentration measured by HPLC and the activities of Na+ -K+ -ATPase, Ca2+ ATPase and SOD assayed by kit method. RESULTS: Both IFN-gamma and TNF-gamma could decrease sperm acrosin activity and acrosome reaction (P < 0.05 or P < 0.01). TNF alpha showed stronger inhibiting effect, IFN-gamma markedly reduced the activities of Na+ -K+ -ATPase, Ca2+ -ATPase and SOD in sperm (P < 0.01), and their synergistic action was weaker. However TNF-alpha produced hardly any effect on Na+ -K+ -ATPase and Ca2+ -ATPase. The NO concentration in sperm was significantly increased by IFN-gamma and/or TNF-alpha (P < 0.01). CONCLUSION: IFN gamma and TNF-alpha have some inhibiting effect on sperm acrosin activity and the rate of acrosome reaction, which could be attributed to their influence on the activities of Na+ -K+ -ATPase, Ca2+ -ATPase and SOD, the NO concentration and so on. PMID- 17918704 TI - [Expressions of endothelin-1 in benign prostatic hyperplasia and prostate cancer and their significance]. AB - OBJECTIVE: To investigate the different expressions of endothelin-1 ET-1) in prostate cancer (PCa) and benign prostatic hyperplasia (BPH) tissues and their clinical significance. METHODS: A total of 36 BPH and 44 PCa specimens were examined for the expression of ET-1 by immunohistochemical technique (Elivision plus method). The staining intensity for ET-1 was assessed by light microscopy on a scale from "-" to "+ + +". RESULTS: Positive immunoreactivity was found in BPH and PCa, with a positive rate of 100%. Positive staining was located mostly in the cytoplasm of glandular epithelia and smooth muscle cells of both BPH and PCa and was noted in all stroma vascular endothelial cells. These were no significant differences in the intensity of positive staining for ET-1 between the groups of BPH and PCa (P > 0.05), bone metastasis (BM) and non-BM (P > 0.05), and highly and moderately differentiated PCa (P > 0.05), but the staining intensity for ET-1 was significantly higher in the poorly than in the highly and moderately differentiated PCa (P < 0.01). CONCLUSION: ET-1 has a high expression and the localization is the same in both BPH and PCa. It is involved in the development and progression of BPH and PCa. PMID- 17918705 TI - [Influence of varicocele on sperm chromatin structure and sperm motility]. AB - OBJECTIVE: To study the influence of varicocele on sperm chromatin structure and sperm motility. METHODS: Routine semen analysis and sperm chromatin structure assay (SCSA) were performed in a varicocele group (n=74) and a control group (n=89). RESULTS: Sperm concentration (41.4 +/- 38.7] x 10(6)/ml) grade a+b sperm percentage ([31.7 +/- 16.9]% and sperm viability ([62.8 +/- 22.2]%) in the varicocele group were evidently lower than those ([80.9 +/- 63.1] x 10(6)/ml, [46.8 +/- 20.5]%, [77.2 +/- 17.5])% in the control group (P < 0.05) and so were VCL, VSL and VAP ([37.4 +/- 12.5 microm/s, [23.4 +/- 7.8] microm/s, [26.5 +/- 8.2] microm/s) in the varicocele group than those ([42.4 +/- 10.7] microm/s, [27.3 +/- 7.3] microm/s, [30.7 +/- 7.8] microm/s) in the control (P < 0.05). MAD was increased (P < 0.01), and the COMP alphat of SCSA (23.2 +/-16.2) was obviously higher in the former than in the latter (14.1 +/- 11.8) (P < 0.05). CONCLUSION: Varicocele causes damage to sperm DNA and changes sperm motility, which may result in male infertility. PMID- 17918706 TI - [Transrectal ultrasound: an applicable diagnostic approach to chronic prostatitis]. AB - OBJECTIVE: To assess the application value of transrectal ultrasound (TRUS) in the diagnosis of chronic prostatitis. METHODS: TRUS and examination of prostatic secretion (EPS) were used in the diagnosis of 3 500 cases of chronic prostatitis from September, 2000 to May, 2006. RESULTS: Lower resonance of the inner gland, low-level echo, uneven echo light spots, incomplete outlines and unsmooth borderlines were found in 2279 cases (65.1%), and the enlarged prostate in 1 084 cases (31.0%), with clear integrated amicula and enhanced echogenic spots at the juncture of the external and inner gland. No obvious changes were noted in 137 cases (4.0%), and in another 391 cases (11.2%) were detected alteration of the acoustic image of cystospermitis and blurred margins and uneven echoes of the seminal vesicle. The WBC count in EPS was < 10/HP in 132 cases (3.8%), 10-19/HP in 2 156 cases (61.6%) and > or =20/HP in 1212 cases (34.6%). CONCLUSION: TRUS, as a diagnostic means for chronic prostatitis, can be easily performed and causes little pain and therefore is readily accepted by patients. Combined with EPS, TRUS can provide more definite diagnostic evidence, and for those who are afraid of pain and reject EPS, it is a desirable alternative in the diagnosis of chronic prostatitis. PMID- 17918707 TI - [Expression of ARA55 mRNA in prostate carcinoma tissues and its significance]. AB - OBJECTIVE: To investigate the expression of ARA55 mRNA in the prostate carcinoma tissues and its clinical significance. METHODS: Real-time RT-PCR was used to examine the expression of ARA55 mRNA in the samples of the prostate carcinoma tissues from 32 patients. RESULTS: ARA55 mRNA expressed in different degrees in all the samples. The Ct values were 20.57 +/- 0.20 and 16.33 +/- 0.31 at T1-T2 and T3-T4 stages, 23.13 +/- 0.13 and 17.3 +/- 0.19 for those with Gleason score < or =7 and >7, 24.70 +/- 0.27 and 17.21 +/- 0.34 for those with PSA < or =10 microg/L and >10 microg/L, and 23.82 +/- 0.21 and 16.71 +/- 0.32 for those that responded to endocrinological therapy and those that failed to, respectively. There was a significant difference between the former and the latter. CONCLUSION: ARA55 mRNA expression is significantly correlated with the clinical characteristics of the patient. And ARA55 can be regarded as a prognostic molecular marker for prostate carcinoma as well as a predictor of endocrinological therapeutic effect on the disease. PMID- 17918708 TI - [Expression of augmenter of liver regeneration in cryptorchidism spermatogenic cells and its implication]. AB - OBJECTIVE: To investigate the expression of the augmenter of liver regeneration (ALR) in cryptorchidism spermatogenic cells. METHODS: Twenty cryptorchidism rat models were established by surgery, 10 included as normal controls and anther 10 as sham-surgery controls. The expression of ALR was detected by immunochemistry, the COX II level measured by immunofluorescence and the ferric iron contents assayed by Perls stain. RESULTS: ALR expressed intensively in the spermatogonia of the control groups but in a signigicantly diminished manner in the cryptorchidism group. No significant difference was found in the COX II level between any two groups of the same age. Ferric iron content of the PND30 cryptorchids decreased significantly (P < 0.05). CONCLUSION: ALR may play an important role in early spermatogenesis. Metabolism dysfunctions caused by ALR defection might be a crucial mechanism for aspermatogenesis of cryptorchidism. PMID- 17918709 TI - [Changes in the expression of voltage-dependent ca2+ channels in asthenospermia]. AB - OBJECTIVE: To study the relationship between the expression of voltage-dependent Ca2+ channels (VDCCs) cal mRNA and asthenospermia. METHODS: Based on the WHO criteria, we filtered the donated semen by computer-aided sperm analysis (CASA), optimized the ejaculated sperm by discontinuous Percoll grads centrifugation, and examined and quantitated the multitype VDCCs alpha1 mRNA expressions in the sperm of normal men and asthenospermia patients using reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Compared with the normal sperm, there were significant differences among the expressions of alpha1B (39.40 +/- 9.47), alpha1C (48.30 +/- 11.60), alpha1E (3.20 +/- 0.78), alpha1G (8.40 +/- 2.03) and alpha1H (5.70 +/- 1.47) mRNA messages in asthenospermia (P < 0.01). CONCLUSION: The abnormal expression of L-type and/or non-L-type VDCCs' mRNAs may be one of the causes of asthenospermia. PMID- 17918710 TI - [Effects of transurethral catheterization on uroflow rate in the pressure-flow study of patients with benign prostatic hyperplasia]. AB - OBJECTIVE: To investigate the effects of transurethral catheterization on the uroflow rate in the pressure-flow study of patients with benign prostatic hyperplasia (BPH). METHODS: Thirty-nine men with BPH underwent free uroflowmetry and pressure-flow analysis respectively. With an 8 F urethral catheter, the standard pressure-flow variables such as the maximum flow rate, detrusor pressure at the maximum flow rate and so on were recorded. The free maximum flow rate and the maximum flow rate with transurethral catheterization were statistically analyzed. RESULTS: The maximum voided volumes of the free uroflowmetry group and the pressure-flow study group were (209.23 +/- 56. 56) ml and (210.33 +/- 62.02) ml respectively (P > 0.05). The free maximum flow rate was (8.61 +/- 2.80) ml/s, and the maximum flow rate with transurethral catheterization-was (7.39 +/- 3.01) ml/s (P < 0.05). When the patients were divided into seven grades of bladder outlet obstruction (BOO) according to the Schafer nomogram, the free maximum flow rate and the maximum flow rate with transurethral catheterization with Grade 0-I of BOO were (12.56 +/- 1.57) ml/s and (10.95 +/- 2.51) ml/s, and those of Grade II were (9.35 +/- 0.76) ml/s and (8.41 +/- 1.23) ml/s respectively. For Grades III, IV and V-VI , the two maximum flow rates were (7.88 +/- 1.21) ml/s and (6.37 +/- 0.59) ml/s, (6.54 +/- 1.93) ml/s and (5.55 +/- 2.48) ml/s, and (6.01 +/- 2.10) ml/s and (4.84 +/- 2.89) ml/s, respectively, all with significant difference in between (P < 0.05). CONCLUSION: The 8 F urethral catheter has a significant effect on the maximum uroflow rate in the pressure-flow study and this effect is correlated with the grade of BOO. PMID- 17918711 TI - [Autosomal DAZL single nucleotide polymorphisms not associated with male infertility in northeast China]. AB - OBJECTIVE: To investigate the association of male infertility with single nucleotide polymorphisms of the autosomal DAZL gene. METHODS: Semen samples were collected from infertile men (infertility group, n=144) and healthy fathers (fertility group, n=53) in the northeast of China, and the former was further divided into 5 subgroups according to the WHO diagnostic criteria for oligoasthenoteratozoospermia (1999) i.e., oligoasthenoteratozoospermia, oligoasthenozoospermia, asthenoteratozoospermia, asthenozoospermia and non oligoasthenoteratozoospermia. PCR-RFLP analysis was used to detect the polymorphism of SNP260. The SNP386 polymorphism of the DAZL gene was analysed with a PCR-SSCP based method. Sequencing and statistical analyses were performed. RESULTS: The SNP260 polymorphism of the DAZL gene was found in both the infertility and the control groups, with no significant difference in between (P > 0.05). There was a larger percentage of SNP260AG in the subgroup of oligoasthenoteratozoospermia than in others, and the SNP386 polymorphism of the DAZL gene was not found in any of the subgroups. CONCLUSION: The SNP260 and SNP386 polymorphisms of the DAZL gene are not associated with male infertility in the northeast of China. Further research needs to be done to clarify the association between SNP260AG and oligoasthenoteratozoospermia. SNP386 polymorphism may be restricted in some small area in Taiwan only. Therefore neither of them can be used as a molecular marker for the genetic diagnosis of male infertility in the northeast of China. PMID- 17918712 TI - [An fMRI study on brain activation patterns of males and females during video sexual stimulation]. AB - OBJECTIVE: To investigate the difference in the brain activation patterns of males and females during video sexual stimulation by functional magnetic resonance imaging (fMRI). METHODS: The participants were 20 adult males and 20 adult females, all healthy, right-handed, and with no history of sexual function disorder and physical, psychiatric or neurological diseases. Blood-oxygen-level dependent fMRI was performed using a 1.5 T MR scanner. Three-dimensional anatomical image of the entire brain were obtained by using a T1-weighted three dimensional anatomical image spoiled gradient echo pulse sequence. Each person was shown neutral and erotic video sequences for 60 s each in a block-study fashion, i.e. neutral scenes--erotic scenes--neutral scenes, and so on. The total scanning time was approximately 7 minutes, with a 12 s interval between two subsequent video sequences in order to avoid any overlapping between erotic and neutral information. RESULTS: The video sexual stimulation produced different results in the men and women. The females showed activation both in the left and the right amygdala, greater in the former than in the latter ([220.52 +/- 17.09] mm3 vs. [155.45 +/- 18.34] mm3, P < 0.05), but in the males only the left amygdala was activated. The males showed greater brain activation than the females in the left anterior cingulate gyrus ([420.75 +/- 19.37] mm3 vs. [310.67 +/- 10.53] mm3, P < 0.05), but less than the females in the splenium of the corpus callosum ([363.32 +/- 13.30] mm3 vs. [473.45 +/- 14.92] mm3, P < 0.01). CONCLUSION: Brain activation patterns of males and females during video sexual stimulation are different, underlying which is presumably the difference in both the structure and function of the brain between men and women. PMID- 17918713 TI - [Inhibition of telomerase with hTERT antisense enhances TNF-alpha-induced apoptosis in prostate cancer cells PC3]. AB - OBJECTIVE: To investigate the inhibiting effect of telomerase with hTERT antisense on TNF-alpha-induced apoptosis in prostate cancer cells PC3. METHODS: Antisense phosphorothioate oligodeoxynucleotide (AS PS-ODN) was synthesized and purified. Telomerase activity was measured by telomeric repeat amplification protocol (TRAP) and telomerase PCR-ELISA Kit, cell viability was determined by MTT assay, and cell apoptosis was observed by morphological method and determined by flow cytometry. RESULTS: AS PS-ODN could significantly inhibit the telomerase activity and increase the susceptibility of TNF-alpha-induced apoptosis of PC3 cells. CONCLUSION: Inhibition of telomerase with hTERT antisense can enhance TNF alpha-induced apoptosis in prostate cancer cells. PMID- 17918714 TI - [Pelvic vas deferens abscess: a case report and review of the literature]. AB - OBJECTIVE: To investigate the clinical characteristics, diagnosis, misdiagnosis and treatment of pelvic vas deferens abscess. METHODS: The clinical data of one case of pelvic vas deferens abscess were reported and the relevant literature was reviewed. RESULTS: The patient underwent incision and drainage of pelvic vas deferens abscess. After the catheter was removed, he could urinate easily. With a follow-up of one year, he was found free of dysuria and recurrence. CONCLUSION: Pelvic vas deferens abscess tends to be misdiagnosed when enwrapped. Surgical procedures are the most effective treatment for this disease. PMID- 17918715 TI - [Sonic hedgehog and prostate growth regulation]. AB - Sonic hedgehog (Shh) is a key signal regulatory factor in embryonic development. It is reported that Shh signaling plays important roles in prostatic duct differentiation and matrix-epithelium interaction, and thus regulates the development, growth and cell proliferation of the prostate. A disorder in Shh signaling will lead to the production and proliferation of tumor cells. An exploration into the mechanism of Shh signaling in the normal growth and diseased condition of the prostate will offer some significant ideas for the studies on the pathogenesy of prostate diseases. PMID- 17918716 TI - [Cryopreservation of spermatozoa]. AB - Sperm cryopreservation has revolutionized the field of assisted reproduction. However, it causes cryodamage to the structure and function of spermatozoa during the freezing-thawing process. Optimization of sperm cryopreservation is necessary for the preservation of male fertility. Cryodamage can be reduced effectively by such methods as improvement of semen quality before freezing, spermatozoal selection, addition of optimal cryoprotectants and application of appropriate thawing techniques. Recent studies focus on cryobiology, improvement of freezing thawing methods, especially for poor quality semen, and evaluation criteria for post-thaw spermatozoa. PMID- 17918717 TI - [Advances in minimally invasive treatment of benign prostatic hyperplasia]. AB - The therapeutic options for benign prostatic hyperplasia (BPH) can be divided into medicinal, surgical and minimally invasive treatments. The minimally invasive treatment of BPH is gradually accepted by more and more urologists for its advantages of less damage, good effect, quick recovery and easy acceptance by patients, which includes transurethral needle ablation of the prostate (TUNA), transurethral microwave therapy (TUMT), photoselective laser vaporization of the prostate (PVP), holmium laser enucleation of the prostate (HOLEP) and so on. This article updates the advances in the minimally invasive treatment. PMID- 17918718 TI - [Pneumoscrotum induced by spontaneous colon perforation: a case report and review of the literature]. AB - One case of pneumoscrotum associated with spontaneous colon perforation was reported. The patient was a 66-year-old man, presented with high temperature, mild abdominal pain and an enlarged scrotum. Physical examination revealed scrotal swelling, abdominal tenderness Case Report and muscular defense. Computed tomography (CT) of the abdomen showed swelling and pneumatosis of the left major psoas and iliopsoas muscles, and ultrasound found subcutaneous emphysema of the scrotum. Surgical investigation discovered a retroperitoneal perforation in the descending colon connected with a huge retroperitoneal vomica and scrotal sac. Spontaneous colon perforation induced pneumoscrotum is rare clinically. It may present as colon perforation, which calls for special attention. PMID- 17918719 TI - [Kidney-jing deficiency reduces the fertility of male mice and their male offspring]. AB - OBJECTIVE: To investigate the effects of kidney-jing deficiency on the fertility of male mice and their male offspring. METHODS: Thirty 6-week-old Kunming male mice and 300 female ones were randomly allocated to a blank control group, a model group and a kidney-tonifying group. The model and the kidney-tonifying groups were stressed by fear plus excessive sex to establish a kidney-jing deficiency model, and meanwhile the latter were given concentrated solution of Kidney-tonifying Recipe intragastrically at the dose of 0.16 ml/10 g. The control and the model groups were treated with physiological saline at the same dose for 21 days. Then all the male mice were mated with the healthy estrous females for 5 days. The sperm density and motility of each group of the male mice were examined, and their fertility was assessed by comparison of the pregnancy rate and the number of baby mice at each birth among their female mates. And the sperm density and motility of the male offspring were detected at 6 weeks. RESULTS: The average number of baby mice at each birth in the model group was (7.00 +/- 1.73), significantly smaller than those in the control (9.43 +/- 1.27) and the kidney tonifying group (8.80 +/- 1.10) (P < 0.05). The sperm density and motility of the model mice were (9.70 +/- 1.15) x 10(6) / ml and (66.72 +/- 10. 12) %, lower than those of the control ([14.08 +/- 1.15 x 10(6)/ ml and [81.75 +/- 3.56] %), and the kidney-tonifying group ([12.20 +/- 1.55] x 10(6)/ ml and [78.55 +/- 4.38] %) (P < 0.05). There was no significant difference between the latter two groups (P > 0.05). The sperm density and motility of the offspring of the model mice were (10.10 +/- 1.79) x 10(6)/ ml and (71.86 +/- 7.48) %, lower than those of the control ([15.30 +/- 1.83] x 10(6)/ ml and [79.86 +/- 5.68] %), and the kidney tonifying group ([14.20 +/- 2.21] x 10(6)/ ml and [81.92 +/- 2.51] %) (P < 0.05), with no significant difference between the latter two groups (P > 0.05). CONCLUSION: Fear plus excessive sex could reduce the fertility of male mice and even that of their male offspring. And kidney-tonifying therapy could counteract this effect. PMID- 17918720 TI - [The safety of tadalafil in the treatment of erectile dysfunction]. AB - The cyclic nucleotide (cGMP) signalling pathway mediates the smooth-muscle relaxing effects of nitric oxide necessary for normal erectile function. Down regulation of this pathway is the pathophysiological pivot of many forms of erectile dysfunction (ED) and leads to the development of some chronic diseases, such as hypertension and type 2 diabetes mellitus. Therefore, selective inhibition of the enzyme that catalyses the degradation of cGMP promotes erectile responses to sexual stimulation. Recently, a new phosphodiesterase type 5 (PDE-5) inhibitor tadalafil has emerged, which has a prolonged half-life. Here is a review of recent studies on the safety of tadalafil in the treatment of ED. PMID- 17918722 TI - True density analysis of a freeze-dried amorphous sugar matrix. AB - True density of an amorphous matrix represents the state of molecular packing in the matrix, which is closely related to the physical/chemical properties of the material. Dry gas pycnometry is one possible technique for measuring the true density of an amorphous sugar matrix prepared by freeze-drying. We herein report on the influence of conditions used for pycnometry on the measured density value and propose a protocol for obtaining the true density. The technique is sufficiently accurate to permit values for matrices comprised of different types of sugar to be compared. Using the protocol, the true densities of several amorphous sugar samples containing different types of sugar, freeze-drying conditions (temperature and sugar concentration at the time of freezing of an aqueous sugar solution), pretreatment (compaction and grind) were determined and the results were compared. A model for simulating an amorphous matrix of sugar (trehalose) was constructed using molecular dynamics/mechanics calculations, and the true density of the simulated sugar matrix was found to agree with the value experimentally determined using the proposed protocol. The relationship among the true density, the states of intermolecular interactions, and strain of sugar molecules in the matrix are discussed using the simulated amorphous sugar matrix. PMID- 17918724 TI - Detecting and identifying the complexation of nimodipine with hydroxypropyl-beta cyclodextrin present in tablets by Raman spectroscopy. AB - For CD-based formulations, it is important to directly monitor the complexation status of the drug present in final dosage form pharmaceuticals. In this work, Raman spectroscopy was explored for the detection and identification of the Nimodipine/hydroxypropyl-beta-cyclodextrin (NMD/HPbetaCD) complexation present in the tablet. The evident, consistent Raman spectral changes in the shift, height ratio and area ratio for the characteristic bands of NMD molecule were featured in the NMD/HPbetaCD complex, and employed to distinguish between complexed and uncomplexed NMD in tablets. A number of practical issues for the Raman measurements performed on the tablets were considered and addressed. The Raman approach and dissolution test were applied to different tablets prepared experimentally at variable granulations. The results demonstrated that the Raman approach can serve as a promising methodology for tablet identification on the complexation. Wet granulation facilitated the process-induced transformations in the complexation. The adequate ethanol in the granulating fluid appeared optimized for the complexation of the two components. The spectral characteristics for dissociation in the tablets were in full accordance with the observations of their diminished initial dissolution parameters. It implied the possibility that the tablet dissolution can be predicted from the Raman interpretation. PMID- 17918725 TI - Molecular design for enhancement of ocular penetration. AB - Over the past two decades, many oral drugs have been designed in consideration of physicochemical properties to attain optimal pharmacokinetic properties. This strategy significantly reduced attrition in drug development owing to inadequate pharmacokinetics during the last decade. On the other hand, most ophthalmic drugs are generated from reformulation of other therapeutic dosage forms. Therefore, the modification of formulations has been used mainly as the approach to improve ocular pharmacokinetics. However, to maximize ocular pharmacokinetic properties, a specific molecular design for ocular drug is preferable. Passive diffusion of drugs across the cornea membranes requires appropriate lipophilicity and aqueous solubility. Improvement of such physicochemical properties has been achieved by structure optimization or prodrug approaches. This review discusses the current knowledge about ophthalmic drugs adapted from systemic drugs and molecular design for ocular drugs. I propose the approaches for molecular design to obtain the optimal ocular penetration into anterior segment based on published studies to date. PMID- 17918726 TI - Docking and scoring protein complexes: CAPRI 3rd Edition. AB - The performance of methods for predicting protein-protein interactions at the atomic scale is assessed by evaluating blind predictions performed during 2005 2007 as part of Rounds 6-12 of the community-wide experiment on Critical Assessment of PRedicted Interactions (CAPRI). These Rounds also included a new scoring experiment, where a larger set of models contributed by the predictors was made available to groups developing scoring functions. These groups scored the uploaded set and submitted their own best models for assessment. The structures of nine protein complexes including one homodimer were used as targets. These targets represent biologically relevant interactions involved in gene expression, signal transduction, RNA, or protein processing and membrane maintenance. For all the targets except one, predictions started from the experimentally determined structures of the free (unbound) components or from models derived by homology, making it mandatory for docking methods to model the conformational changes that often accompany association. In total, 63 groups and eight automatic servers, a substantial increase from previous years, submitted docking predictions, of which 1994 were evaluated here. Fifteen groups submitted 305 models for five targets in the scoring experiment. Assessment of the predictions reveals that 31 different groups produced models of acceptable and medium accuracy-but only one high accuracy submission-for all the targets, except the homodimer. In the latter, none of the docking procedures reproduced the large conformational adjustment required for correct assembly, underscoring yet again that handling protein flexibility remains a major challenge. In the scoring experiment, a large fraction of the groups attained the set goal of singling out the correct association modes from incorrect solutions in the limited ensembles of contributed models. But in general they seemed unable to identify the best models, indicating that current scoring methods are probably not sensitive enough. With the increased focus on protein assemblies, in particular by structural genomics efforts, the growing community of CAPRI predictors is engaged more actively than ever in the development of better scoring functions and means of modeling conformational flexibility, which hold promise for much progress in the future. PMID- 17918727 TI - Assessing the reliability of sequence similarities detected through hydrophobic cluster analysis. AB - Hydrophobic cluster analysis (HCA) has long been used as a tool to detect distant homologies between protein sequences, and to classify them into different folds. However, it relies on expert human intervention, and is sensitive to subjective interpretations of pattern similarities. In this study, we describe a novel algorithm to assess the similarity of hydrophobic amino acid distributions between two sequences. Our algorithm correctly identifies as misattributions several HCA-based proposals of structural similarity between unrelated proteins present in the literature. We have also used this method to identify the proper fold of a large variety of sequences, and to automatically select the most appropriate structure for homology modeling of several proteins with low sequence identity to any other member of the protein data bank. Automatic modeling of the target proteins based on these templates yielded structures with TM-scores (vs. experimental structures) above 0.60, even without further refinement. Besides enabling a reliable identification of the correct fold of an unknown sequence and the choice of suitable templates, our algorithm also shows that whereas most structural classes of proteins are very homogeneous in hydrophobic cluster composition, a tenth of the described families are compatible with a large variety of hydrophobic patterns. We have built a browsable database of every major representative hydrophobic cluster pattern present in each structural class of proteins, freely available at http://www2.ufp.pt/ pedros/HCA_db/index.htm. PMID- 17918721 TI - Past, present, and future technologies for oral delivery of therapeutic proteins. AB - Biological drugs are usually complex proteins and cannot be orally delivered due to problems related to degradation in the acidic and protease-rich environment of the gastrointestinal (GI) tract. The high molecular weight of these drugs often results in poor absorption into the periphery when administered orally. The most common route of administration for these therapeutic proteins is injection. Most of these proteins have short serum half-lives and need to be administered frequently or in high doses to be effective. So, difficulties in the administration of protein-based drugs provides the motivation for developing drug delivery systems (DDSs) capable of maintaining therapeutic drug levels without side effects as well as traversing the deleterious mucosal environment. Employing a polymer as an entrapment matrix is a common feature among the different types of systems currently being pursued for protein delivery. Protein release from these matrices can occur through various mechanisms, such as diffusion through or erosion of the polymer matrix, and sometimes a combination of both. Encapsulation of proteins in liposomes has also been a widely investigated technology for protein delivery. All of these systems have merit and our worthy of pursuit. PMID- 17918728 TI - Progress from CASP6 to CASP7. AB - This article describes the general quality of models of three dimensional structure submitted to CASP7 and analyzes progress since the previous experiment, primarily using measures that were used in earlier analyses. Overall improvement in model accuracy compared to CASP6 is modest, but there are two developments of note: server performance has moved closer to that of humans, and there has been a significant improvement in the fraction of targets for which the best model is superior to that obtainable using knowledge of a single best template structure. PMID- 17918730 TI - Hydrophobic interactions and ionic networks play an important role in thermal stability and denaturation mechanism of the porcine odorant-binding protein. AB - Despite the fact that the porcine odorant-binding protein (pOBP) possesses a single tryptophan residue (Trp 16) that is characterized by a high density microenvironment (80 atoms in a sphere with radius 7 A) with only one polar group (Lys 120) and three bound water molecules, pOBP displayed a red shifted fluorescence emission spectrum (lambda(max) = 340 nm). The protein unfolding in 5M GdnHCl was accompanied by the red shift of the fluorescence emission spectrum (lambda(max) = 353 nm), by the increase of fluorescence quantum yield, and by the decrease of lifetime of the excited state (from 4.25 ns in native state to 3.15 ns in the presence of 5M GdnHCl). Taken together these data indicate the existence of an exciplex complex (Trp 16 with Lys 120 and/or with bound molecules of water) in the protein native state. Heat-induced denaturation of pOBP resulted in significant red shifts of the fluorescence emission spectra: the value of the ratio (I(320)/I(365)) upon excitation at lambda(ex) = 297 nm (parameter A) decreases from 1.07 to 0.64 passing from 60 to 85 degrees C, and the calculated midpoint of transition was centered at 70 degrees C. Interestingly, even at higher temperature, the values of the parameter A both in the absence and in the presence of GdnHCl did not coincide. This suggests that a portion of the protein structure is still preserved upon the temperature-induced denaturation of the protein in the absence of GdnHCl. CD experiments performed on pOBP in the absence and in the presence of GdnHCl and at different temperatures were in agreement with the fluorescence results. In addition, the obtained experimental data were corroborated by the analysis of the 3D structure of pOBP which revealed the amino acid residues that contribute to the protein dynamics and stability. Finally, molecular dynamics simulation experiments pointed out the important role of ion pair interactions as well as the molecular motifs that are responsible for the high thermal stability of pOBP, and elucidated the reasons of the protein aggregation that occurred at high temperature. PMID- 17918729 TI - Critical assessment of methods of protein structure prediction-Round VII. AB - This paper is an introduction to the supplemental issue of the journal PROTEINS, dedicated to the seventh CASP experiment to assess the state of the art in protein structure prediction. The paper describes the conduct of the experiment, the categories of prediction included, and outlines the evaluation and assessment procedures. Highlights are improvements in model accuracy relative to that obtainable from knowledge of a single best template structure; convergence of the accuracy of models produced by automatic servers toward that produced by human modeling teams; the emergence of methods for predicting the quality of models; and rapidly increasing practical applications of the methods. PMID- 17918731 TI - Liposomal delivery of hydrophobic weak acids: enhancement of drug retention using a high intraliposomal pH. AB - Clinical development of highly potent lipophilic neutral camptothecins has been impeded by the poor solubility, stability, and nonspecific toxicity of these compounds. Liposomal encapsulation offers a promising formulation route for tumor site-specific delivery of these novel drug candidates. However, the development of formulation strategies for liposomal loading and retention of hydrophobic drugs such as the neutral camptothecins has been lacking. In the studies presented here, we explored the potential of a trans-bilayer pH gradient strategy for prolonging the liposome retention of DB-67, a novel lipophilic camptothecin that can undergo lactone ring-opening to form a hydrophobic weak acid. The liposome membrane permeability of DB-67 was obtained as a function of pH in aqueous buffers. A permeability model was developed and liposome membrane permeability was shown to be controlled by the fraction of unbound neutral lactone entrapped in the vesicles. Liposome membrane permeability of DB-67 was also studied under physiological conditions. The high membrane partitioning of DB 67 in the intraliposomal microenvironment was found to shift the equilibrium between lactone and carboxylate towards the lactone species resulting in a faster than desired drug release under physiological conditions. The effectiveness of the pH gradient strategy was further reduced under physiological conditions by the rapid loss of trans-membrane pH gradients due to CO(2) uptake. Simulations were conducted to explore the role of membrane binding, intravesicular pH, and carbonate buffer concentration in successful utilization of the pH gradient strategy for hydrophobic weak acids. PMID- 17918732 TI - An integrated genetic and functional analysis of the role of type II transmembrane serine proteases (TMPRSSs) in hearing loss. AB - Building on our discovery that mutations in the transmembrane serine protease, TMPRSS3, cause nonsyndromic deafness, we have investigated the contribution of other TMPRSS family members to the auditory function. To identify which of the 16 known TMPRSS genes had a strong likelihood of involvement in hearing function, three types of biological evidence were examined: 1) expression in inner ear tissues; 2) location in a genomic interval that contains a yet unidentified gene for deafness; and 3) evaluation of hearing status of any available Tmprss knockout mouse strains. This analysis demonstrated that, besides TMPRSS3, another TMPRSS gene was essential for hearing and, indeed, mice deficient for Hepsin (Hpn) also known as Tmprss1 exhibited profound hearing loss. In addition, TMPRSS2, TMPRSS5, and CORIN, also named TMPRSS10, showed strong likelihood of involvement based on their inner ear expression and mapping position within deafness loci PKSR7, DFNB24, and DFNB25, respectively. These four TMPRSS genes were then screened for mutations in affected members of the DFNB24 and DFNB25 deafness families, and in a cohort of 362 sporadic deaf cases. This large mutation screen revealed numerous novel sequence variations including three potential pathogenic mutations in the TMPRSS5 gene. The mutant forms of TMPRSS5 showed reduced or absent proteolytic activity. Subsequently, TMPRSS genes with evidence of involvement in deafness were further characterized, and their sites of expression were determined. Tmprss1, 3, and 5 proteins were detected in spiral ganglion neurons. Tmprss3 was also present in the organ of Corti. TMPRSS1 and 3 proteins appeared stably anchored to the endoplasmic reticulum membranes, whereas TMPRSS5 was also detected at the plasma membrane. Collectively, these results provide evidence that TMPRSS1 and TMPRSS3 play and TMPRSS5 may play important and specific roles in hearing. PMID- 17918733 TI - Liposomal phospholipid preparations of chloramphenicol for ophthalmic applications. AB - Since their discovery by Bangham and coworkers almost four decades ago, liposomes have become models for biomembranes and vehicles for pharmaceutical, diagnostic, and cosmetic agents. One of the advantages of using liposomes as a drug vehicle is their ability for slow release, thus reducing dosage, localizing a drug, and minimizing its side-effects. Antibiotic resistance is a growing global problem, including for ocular bacterial infection by Staphylococcus aureus, where time is an important parameter that determines the severity of infection. This situation has prompted the pursuit of ways to prepare drug-encapsulating liposomes that enable bulk release of the drug chloramphenicol (CAP) once the liposomal structure is perturbed. Our approach is a two-step process: first, to characterize the interaction of CAP with model biomembranes of dimyristoylphosphatidylcholine (DMPC) used to prepare CAP-liposomes by different formulations; second, to test the efficiency of these formulation against S. aureus. Solid-state NMR, differential scanning calorimetry, and infrared spectroscopy were used to study the interaction of CAP with DMPC bilayers. The minimum inhibitory concentrations and time-kill curves for S. aureus using different liposomal preparations were compared. Evidence of conformational changes in the DMPC molecules and the effectiveness of the CAP encapsulated in liposomes are reported. PMID- 17918734 TI - Monosomy 1p36 deletion syndrome. AB - Monosomy 1p36 results from a heterozygous deletion of the most distal chromosomal band on the short arm of chromosome 1. Occurring in approximately 1 in 5,000 live births, monosomy 1p36 is the most common terminal deletion observed in humans. Monosomy 1p36 is associated with mental retardation, developmental delay, hearing impairment, seizures, growth impairment, hypotonia, and heart defects. The syndrome is also characterized by several distinct dysmorphic features, including large anterior fontanels, microcephaly, brachycephaly, deep-set eyes, flat nose and nasal bridge, and pointed chin. Several genes have been proposed as causative for individual features of the phenotype. In addition, based upon molecular characterization of subjects with monosomy 1p36, several mechanisms for the generation and stabilization of terminal deletions have been proposed. PMID- 17918735 TI - The 6p subtelomere deletion syndrome. AB - Submicroscopic deletion of the 6p25 subtelomere has recently been recognized as a clinically identifiable syndrome. To date, more than 30 cases have been described with variable cytogenetically visible 6p deletions. Terminal 6p deletions result in a clinically distinguishable phenotype. The focus of this review is the phenotype associated with isolated terminal deletions of 6p25, and specifically isolated submiscroscopic subtelomere deletions. A distinct phenotype has emerged consisting of developmental delay/mental retardation, language impairment, hearing loss, and ophthalmologic, cardiac, and craniofacial abnormalities. These features demonstrate considerable clinical overlap with the Ritscher-Schinzel (or cranio-cerebello-cardiac (3C)) syndrome (OMIM #220210). Isolated submiscroscopic 6p25 subtelomere terminal deletion has been reported in 11 individuals, two of whom are siblings. Cytogentic and molecular mapping of the 6p25 deletion boundary has been reported in 8 of these 10 unrelated individuals with isolated submiscroscopic subtelomere deletion. This analysis has revealed substantial phenotypic overlap between individuals with submicroscopic terminal 6p deletions and those with large, cytogenetically visible deletions of the region suggesting that the critical genes contributing to the main clinical and developmental features lie in the terminal region of 6p25. PMID- 17918736 TI - The effect of cyclodextrins on chemical and physical stability of glucagon and characterization of glucagon/gamma-CD inclusion complexes. AB - The purpose of the study was to evaluate the effect of cyclodextrin (CD) complexation on the chemical and physical stability of a polypeptide hormone glucagon and to study the interactions between glucagon and gamma-cyclodextrin molecules in inclusion complexes. The chemical stability of glucagon at pH 2.0 was studied with HPLC-UV and HPLC-MS/MS. The physical stability of glucagon at pH 2.5 was studied by measuring the turbidity (A(405 nm)) and viscosity (Ostwald capillary viscosimeter) of the samples. The structure of glucagon/gamma-CD complexes at pH 2.5 was studied with 2D-NMR. The presence of various CDs increased the chemical half-life of glucagon at pH 2.0 (37 degrees C, 0.01 M HCl, ionic strength 0.15) and prolonged the lag-time before aggregation at pH 2.5 (0.9% (w/v) NaCl in 3.2 mM HCl). The NMR studies showed that the side chains of all the aromatic amino acid residues (Phe6, Tyr10, Tyr13, Phe22, Trp25) and leucines (Leu14 and Leu26) of glucagon interacted with the cavities of the gamma CD molecules. The present study shows that glucagon forms inclusion complexes with cyclodextrins in acidic solution, resulting in an improvement in its chemical and physical stability. PMID- 17918737 TI - Formulating poly(lactide-co-glycolide) particles for plasmid DNA delivery. AB - Biodegradable poly(lactide-co-glycolide) (PLGA) particles have shown significant potential for sustained and targeted delivery of several pharmaceutical agents, including plasmid DNA (pDNA). Here, we survey current approaches to PLGA particle preparation for pDNA delivery and discuss recent progress on optimizing formulation development. PMID- 17918738 TI - Hypoalgesia in mice lacking GABA transporter subtype 1. AB - gamma-Aminobutyric acid (GABA) transporters play a key role in the regulation of GABA neurotransmission. We reported previously that overexpression of the GABA transporter subtype 1 (GAT1), the major form of the GABA transporter in the CNS, led to hyperalgesia in mice. In the present study, nociceptive responses of GAT1 knockout mice (GAT1(-/-)) were compared with those of heterozygous (GAT(+/-)) and wild-type (GAT(+/+)) mice by four conventional pain models (tail-immersion test, hot-plate test, acetic acid-induced abdominal constriction test, and formalin test). In addition, the analgesic effects of two GAT1-selective inhibitors, NO 711 and tiagabine, were examined in all three genotypes using the same four models. Our data demonstrated that GAT1 deficiency because of genetic knockout or acute blockade by selective inhibitors leads to hypoalgesia in mice. These results confirmed the crucial role of GAT1 in the regulation of nociceptive threshold and suggested that GAT1 inhibitors have the potential for clinical use in pain therapy. PMID- 17918740 TI - Dephosphorylation of Akt in C6 cells grown in serum-free conditions corresponds with redistribution of p85/PI3K to the nucleus. AB - Withdrawal of serum from cell cultures constitutes a useful model for the study of mechanisms involved in the regulation of Akt function in vitro. However, there have been several reports of changes in Akt activity that are not fully explained by the current model of phosphatidylinositol 3'-kinase (PI3K)/Akt signaling. We demonstrate the expected loss of Akt phosphorylation in C6 glioma cells cultured in serum-free conditions, yet we also observed a paradoxical increase in PI3K lipid kinase activity in the same cultures. These events corresponded with relocalization of p85, the regulatory subunit of PI3K, to the perinuclear region and a local increase in PI3K-lipid kinase products. Treatment with platelet derived growth factor (PDGF) maintained the association between p85 and the PDGF receptor during serum withdrawal and restored PI3K-lipid production at the plasma membrane. Although this protected Akt from dephosphorylation, it only slightly reversed cell-cycle arrest. These effects were not sensitive to treatment with epidermal growth factor, thus precluding a generalized role for growth factors. Our data suggest that loss of growth factor signaling, including PDGF signaling, may disrupt recruitment and/or anchoring of an active p85(PI3K) complex at the plasma membrane during serum withdrawal, which could account for the concurrent loss of Akt function. PMID- 17918739 TI - Region-specific differentiation of embryonic stem cell-derived neural progenitor transplants into the adult mouse hippocampus following seizures. AB - Embryonic stem (ES) cells can generate neural progenitors and neurons in vitro and incorporate into the adult central nervous system (CNS) following transplantation, suggesting their therapeutic potential for treating neurological disorders. However, our understanding of the conditions that direct ES-derived neural progenitor (ESNP) migration and differentiation within different regions of the adult CNS is incomplete. Rodents treated with the chemoconvulsant kainic acid (KA) experience seizures and display hippocampal sclerosis, as well as enhanced hippocampal neurogenesis, similar to pathological findings in patients with temporal lobe epilepsy (TLE). To examine the potential for ESNPs to incorporate into the adult hippocampus and differentiate into hippocampal neurons or glia following seizure-induced damage, we compared the fates of ESNPs after they were transplanted into the CA3 region or fimbria 1 week following KA-induced seizures. After 4-8 weeks, ESNPs grafted into the CA3 region had migrated to the dentate gyrus (DG), where a small subset adopted neural stem cell fates and continued to proliferate, based on bromodeoxyuridine uptake. Others differentiated into neuroblasts or dentate granule neurons. In contrast, most ESNPs transplanted into the fimbria migrated extensively along existing fiber tracts and differentiated into oligodendrocytes or astrocytes. Hippocampal grafts in mice not subjected to seizures displayed a marked tendency to form tumors, and this effect was more pronounced in the DG than in the fimbria. Taken together, these data suggest that seizures induce molecular changes in the CA3 region and DG that promote region-specific neural differentiation and suppress tumor formation. PMID- 17918741 TI - cAMP-dependent regulation of spinesin/TMPRSS5 gene expression in astrocytes. AB - Spinesin/TMPRSS5 is a mosaic type serine protease that is predominantly expressed in the spinal cord. To identify the mechanism of spinesin expression, we investigated its expression in vivo and in vitro using several cell lines. Immunohistochemical and in situ hybridization analyses revealed that mouse spinesin (m-spinesin) was abundantly expressed in white matter astrocytes. Similarly, we confirmed abundant expression of m-spinesin in astrocyte cell lines. Then, we analyzed the expression of variant forms of m-spinesin in these cell lines. Interestingly, a transmembrane type (type 4) variant was expressed in neuroblastoma and astrocyte cell lines, whereas a cytoplasmic type (type 1) variant was specifically expressed in astrocyte cell lines. Furthermore, expression of both variants was up-regulated by dibutyryl-cAMP (dbcAMP) treatment only in astrocyte cell lines. We also analyzed the promoter region of the m spinesin gene and revealed that the 5'-flanking region from base pairs -224 to 188 was essential for cAMP-dependent regulation of its transcription. These results indicate that m-spinesin is involved in the function of astrocytes in the spinal cord and that there may be astrocyte-specific regulation of its gene expression. PMID- 17918742 TI - Increased GAD67 mRNA expression in cerebellar interneurons in autism: implications for Purkinje cell dysfunction. AB - It has been widely reported that in autism, the number of Purkinje cells (PCs) is decreased, and recently, decreased expression of glutamic acid decarboxylase 67 (GAD67) mRNA in Purkinje cells also has been observed. However, the autism literature has not addressed key GABAergic inputs into Purkinje cells. Inhibitory basket and stellate cell interneurons in the molecular layer of the cerebellar cortex provide direct key GABAergic input into Purkinje cells and could potently influence the output of Purkinje cells to deep cerebellar nuclei. We investigated the capacity for interneuronal synthesis of gamma-amino butyric acid (GABA) in both types of interneurons that innervate the remaining PCs in the posterolateral cerebellar hemisphere in autism. The level of GAD67 mRNA, one of the isoforms of the key synthesizing enzymes for GABA, was quantified at the single-cell level using in situ hybridization in brains of autistic and aged-matched controls. The National Institutes of Health imaging system showed that expression of GAD67 mRNA in basket cells was significantly up-regulated, by 28%, in eight autistic brains compared with that in eight control brains (mean +/- SEM pixels per cell, 1.03 +/ 0.05 versus 0.69 +/- 0.05, respectively; P < 0.0001 by independent t test). Stellate cells showed a trend toward a small increase in GAD67 mRNA levels, but this did not reach significance. The results suggest that basket cells likely provide increased GABAergic feed-forward inhibition to PCs in autism, directly affecting PC output to target neurons in the dentate nucleus and potentially disrupting its modulatory role in key motor and/or cognitive behaviors in autistic individuals. PMID- 17918743 TI - Aggrecan components differentially modulate nerve growth factor-responsive and neurotrophin-3-responsive dorsal root ganglion neurite growth. AB - Aggrecan is one of the major chondroitin sulfate proteoglycans (CSPGs) expressed in the central nervous system. The signaling pathways activated downstream of cell interaction with aggrecan and with CSPGs in general and the importance of chondroitin sulfate-glycosaminoglycan side chains in their inhibition are unclear. Therefore, to analyze the effect of different components of aggrecan in inhibiting neurite growth, neurite outgrowth was quantified in an in vitro model in which chick dorsal root ganglion (DRG) explants were grown on substrates containing aggrecan bound to hyaluronan and link protein as a macromolecular aggregate, aggrecan monomers, hyaluronan, or ChABC-treated aggrecan. Aggrecan aggregate, aggrecan monomer, and hyaluronan inhibited neurite outgrowth from nerve growth factor (NGF)- and neurotrophin-3 (NT3)-responsive DRG neurons. Aggrecan inhibition was dependent on its chondroitin sulfate-glycosaminoglycans, as ChABC digestion alleviated neurite inhibition because of aggrecan. Growth cones displayed full or partial collapse on aggrecan aggregate, hyaluronan, and ChABC-treated aggrecan. Inhibition of Rho kinase (ROCK) with Y27632 increased neurite growth on some but not all of the aggrecan components tested. With NGF in the culture medium, Y27632 increased neurite outgrowth on aggrecan aggregate, monomers, and ChABC-treated aggrecan, but not on hyaluronan. The ROCK inhibitor also increased NT3-responsive outgrowth on aggrecan aggregate and hyaluronan, but not on ChABC-treated aggrecan. This study showed that the matrix proteoglycan aggrecan and its components have multiple effects on neurite outgrowth and that some of these effects involve the Rho/ROCK pathway. PMID- 17918744 TI - Nuclear factor-kappaB decoy amelioration of spinal cord injury-induced inflammation and behavior outcomes. AB - Spinal cord injury (SCI) results in a pathophysiology characterized by multiple locomotor and sensory deficits, resulting in altered nociception and hyperalgesia. SCI triggers an early and prolonged inflammatory response, with increased interleukin-1beta levels. Transient changes are observed in subunit populations of the transcription factor nuclear factor-kappaB (NF-kappaB). There were decreases in neuronal c-Rel levels and inverse increases in p65 and p50 levels. There were no changes in neuronal p52 or RelB subunits after SCI at any time point tested. Similarly, SCI had no effect on oligodendroglial levels of any NF-kappaB subunit. There were significant early increases in COX-2 and inducible nitric oxide synthase mRNA and protein levels after SCI. We used synthetic double stranded "decoy" deoxyoligonucleotides containing selective NF-kappaB protein dimer binding consensus sequences. Decoys targeting the p65/p50 binding site on the COX-2 promoter decreased SCI-induced cell losses, NF-kappaB p65/p50 DNA binding activity, and COX-2 and iNOS protein levels. NF-kappaB p65/p50 targeted decoys improved early locomotor recovery after moderate but not severe SCI, yet ameliorated SCI-induced hypersensitization after both moderate and severe SCI. To determine whether changes in GABA activity played a role in decreased hypersensitivity after SCI and p65/p50 targeted decoy, we counted gamma aminobutyric acid (GABA)-containing neurons in laminae 1-3. There were significantly more GABAergic neurons in the p65/p50 targeted decoy-treated group at the level of injury. PMID- 17918745 TI - Ebselen, a redox regulator containing a selenium atom, induces neurofilament M expression in cultured rat pheochromocytoma PC12 cells via activation of mitogen activated protein kinase. AB - We found that ebselen [2-phenyl-1,2-benzisoselenazol-3(2H)-one] caused phosphorylation of mitogen-activated protein kinase (MAPK), followed by expression of neurofilament-M, a neuron-specific protein, in cultured PC12 rat pheochromocytoma cells. The ebselen-induced MAPK activation was suppressed by U0126, an inhibitor of MAPK kinase (MEK1/2), but not by K252a, a selective inhibitor of Trk family tyrosine kinases; AG1478, an antagonist of epidermal growth factor receptor (EGFR); pertussis toxin, an inhibitor of Gi/o; or GP antagonist-2A, an inhibitor of Gq. Furthermore, we observed that N-acetyl-L cysteine, an inhibitor of tyrosine kinases, suppressed ebselen-induced MAPK activation and buthionine sulfoximine, an activator of protein tyrosine phosphatases, enhanced the effect, indicating that ebselen activated MEK1/2 through one or more tyrosine kinases. Based on these results, we propose that ebselen stimulated intracellular tyrosine kinase activity, thus activating a MAPK cascade (tyrosine kinase-MEK1/2-ERK1/2) in PC12 cells and that this activation resulted in their neuronal differentiation. PMID- 17918747 TI - Developmental expression profile of quaking, a candidate gene for schizophrenia, and its target genes in human prefrontal cortex and hippocampus shows regional specificity. AB - Decreased expression of oligodendrocyte/myelin-related (OMR) genes, including quaking (QKI), is a consistent finding in gene expression studies of post-mortem brain from subjects with schizophrenia, and these changes are most prominent in the hippocampus vs. the prefrontal cortex (PFC). Although expression of QKI and other OMR genes has been examined in rodents, little is known about their developmental trajectory in the human brain. Therefore, we examined expression of QKI and several putative mRNA targets of QKI in human PFC and hippocampus at different ages. The pattern of QKI expression in the PFC resembled that reported in rodents, with high QKI-5 in the fetal brain and an increase in QKI-6 and QKI-7 during the period of active myelination, although QKI-5 expression did not decrease substantially during postnatal development in the PFC in humans as it does in rodent brain. Most of the putative QKI target genes also showed linear increases in expression with increasing age in the PFC. In contrast, expression of these genes showed little evidence of developmental regulation in the hippocampus. Correlations between expression levels of the nuclear vs. cytoplasmic QKI isoforms, and putative splicing targets of the former, also differed between tissues. Thus, we speculate that a robust increase in OMR gene expression normally occurs with age in the PFC, but not in the hippocampus, which may explain why decreases in OMR gene expression in schizophrenia are more pronounced in the latter tissue. We also suggest that OMR transcripts might be processed by different splicing proteins in different tissues. PMID- 17918748 TI - Surrogate therapeutic outcome measures in patients with myasthenia gravis. AB - Treatment of acquired myasthenia gravis (MG) with immunotherapies successfully relieves symptoms and improves strength as documented by the Quantitative Myasthenia Gravis Score for disease severity (QMGS). Neuromuscular function, as demonstrated by the surrogate measures of repetitive nerve stimulation (RNS) and single-fiber electromyography (SFEMG), is sensitive for diagnosis and staging disease severity. This study of 51 patients treated with immunomodulation confirmed that RNS and SFEMG are useful to stage disease severity, but found that clinical measures such as the QMGS are more sensitive to change than electrophysiological parameters. The presence of blocking on SFEMG did predict responsiveness to intravenous immunoglobulin (IVIG) treatment, providing clinicians with an objective, reliable, quantitative measure to help determine which patients will benefit from this costly treatment. PMID- 17918749 TI - Subcutaneous immunoglobulin infusion: a new therapeutic option in chronic inflammatory demyelinating polyneuropathy. AB - Intravenous application of immunoglobulins (IVIg) is an effective and usually well tolerated yet costly therapeutic regimen in chronic inflammatory demyelinating polyneuropathy (CIDP). We report two CIDP patients treated with subcutaneous infusion of immunoglobulins (SCIg) after IVIg therapy was shown to be effective. Application of SCIg was well tolerated, easy to manage, and led to stabilization of the disease course. SCIg may represent an effective new therapeutic option in CIDP and is associated with a cost reduction of at least 50% compared to IVIg therapy. PMID- 17918746 TI - CD40-CD40 ligand interactions in human microglia induce CXCL8 (interleukin-8) secretion by a mechanism dependent on activation of ERK1/2 and nuclear translocation of nuclear factor-kappaB (NFkappaB) and activator protein-1 (AP-1). AB - CXCL8 is a CXC chemokine that recruits leukocytes to sites of inflammation. Expression of CXCL8 in the CNS has been demonstrated in neuroinflammatory diseases, including human immunodeficiency virus (HIV-1) encephalitis, but the mechanism of secretion of this chemokine is not fully understood. CD40 is a 50 kDa protein on the surface of microglia, and we have previously shown that it is increased in expression in HIV-1-infected brain tissue as well as by interferon gamma (IFNgamma) in tissue culture. We examined the expression and regulation of CXCL8 in cultured human fetal microglia after ligation of CD40 with soluble trimeric CD40 ligand (sCD40L) as well as the expression of CXCL8 on microglia in HIV encephalitic brain tissue sections. Treatment of cultured microglia with IFNgamma + sCD40L resulted in significant induction of CXCL8. This expression was mediated by activation of the ERK1/2 MAPK pathway, as demonstrated by ELISA and Western blot using a specific inhibitor (U0126). Gel shift analyses demonstrated that NFkappaB and AP-1, but not C/EBPbeta, mediate microglial CXCL8 production. We also found increased colocalization of CXCL8 with CD68/CD40-positive cells in HIV encephalitic brain tissue compared with HIV-infected nonencephalitic and normal tissue. Thus, CD40-CD40L interactions facilitate chemokine expression, leading to the influx of inflammatory cells into the CNS. These events can lead to the pathology that is associated with neuroinflammatory diseases. PMID- 17918751 TI - The use of circular dichroism spectroscopy for studying the chiral molecular self assembly: an overview. AB - Self-assembly plays an important role in the formation of many chiral biological structures and in the preparation of chiral functional materials. Therefore the control of chirality in synthetic or biological self-assembled systems is important either for the comprehension of recognition phenomena or to obtain materials with predictable and controllable properties. Circular dichroism was developed to study molecular chirality, however, because of its outstanding sensitivity to chiral perturbations of the system under investigation; it has been extended more recently to supramolecular chemistry. In particular, self assembly processes leading to the formation of chiral supramolecular architectures (and eventually to gels or liquid crystal phases) can be monitored by CD. Furthermore, CD spectroscopy often allows one to obtain structural information on the assembled structures. This review deals with representative contributions to the study of supramolecular chirality by means of circular dichroism. PMID- 17918750 TI - Rap2 regulates androgen sensitivity in human prostate cancer cells. AB - BACKGROUND: Progression of prostate cancer to a fatal androgen-independent disease is associated with activation of MAP kinase, consistent with chronic stimulation of the Ras-signaling pathway. We have previously shown that Ras activation is sufficient to induce androgen-independent growth of prostate cancer cells. One mechanism of MAP kinase regulation is modulation of Ras signaling by other Ras family members, the Rap gene paralogs Rap1a/b and Rap2a/b. Here we ask if Rap proteins play a role in determining androgen sensitivity of human prostate cancer cells either alone or in the context of an activated Ras. METHODS: To evaluate the role of Rap proteins in androgen responsiveness we use Rap over expression with or without mutated Ras co-transfection and Rap siRNA knockdown to evaluate androgen-dependent prostate-specific antigen (PSA) promoter reporter expression and cell growth in androgen-dependent LNCaP and independent C4-2 human prostate cancer cells. RESULTS: Rap1 is equally expressed between LNCaP and C4-2 cells and thus we focused on Rap2 which is minimally expressed in C4-2. Rap2a affects androgen-dependent PSA reporter expression in a dose-dependent manner in LNCaP and C4-2 cells. Low levels of Rap2a enhance PSA reporter expression, whereas higher concentrations inhibit expression. We show that Rap2a antagonizes the enhanced PSA reporter expression conferred by an active RasV12 gene in prostate cancer cells. siRNA knockdown data indicate that Rap2 has a greater effect on androgen-stimulated growth in LNCaP than in C4-2 cells. CONCLUSIONS: We show that Rap2 is involved in androgen-mediated transcriptional and growth responses of human prostate cancer cells. PMID- 17918752 TI - Modeling and inference for an ordinal effect size measure. AB - An ordinal measure of effect size is a simple and useful way to describe the difference between two ordered categorical distributions. This measure summarizes the probability that an outcome from one distribution falls above an outcome from the other, adjusted for ties. We develop and compare confidence interval methods for the measure. Simulation studies show that with independent multinomial samples, confidence intervals based on inverting the score test and a pseudo score-type test perform well. This score method also seems to work well with fully-ranked data, but for dependent samples a simple Wald interval on the logit scale can be better with small samples. We also explore how the ordinal effect size measure relates to an effect measure commonly used for normal distributions, and we consider a logit model for describing how it depends on explanatory variables. The methods are illustrated for a study comparing treatments for shoulder-tip pain. PMID- 17918753 TI - Comparison of algorithms to generate event times conditional on time-dependent covariates. AB - The Cox proportional hazards model with time-dependent covariates (TDC) is now a part of the standard statistical analysis toolbox in medical research. As new methods involving more complex modeling of time-dependent variables are developed, simulations could often be used to systematically assess the performance of these models. Yet, generating event times conditional on TDC requires well-designed and efficient algorithms. We compare two classes of such algorithms: permutational algorithms (PAs) and algorithms based on a binomial model. We also propose a modification of the PA to incorporate a rejection sampler. We performed a simulation study to assess the accuracy, stability, and speed of these algorithms in several scenarios. Both classes of algorithms generated data sets that, once analyzed, provided virtually unbiased estimates with comparable variances. In terms of computational efficiency, the PA with the rejection sampler reduced the time necessary to generate data by more than 50 per cent relative to alternative methods. The PAs also allowed more flexibility in the specification of the marginal distributions of event times and required less calibration. PMID- 17918754 TI - Towards configurationally stable [4]helicenes: enantioselective synthesis of 12 substituted 7,8-dihydro[4]helicene quinones. AB - The synthesis of enantiopure C-12 methoxy- or alkyl-substituted 5,7,8,12b tetrahydro[4]helicene quinones 16 and 17 and the 7,8-dihydroaromatic analogues 4 and 5 has been achieved from (SS)-2-(p-tolylsulfinyl)-1,4-benzoquinone. In the first series, with a structure containing both central and helical chiralities, the R absolute configuration of the stereogenic carbon atom was defined after the asymmetric cycloaddition step, whereas the P or M helicity was shown to be dependent on the nature of the C-12 substituent. The size of this group was also defining the configurational stability of the final (P)-7,8-dihydro[4]helicene quinones 4 and 5. The interconversion barriers between the P and M helimers in the latter, computed with a DFT B3LYP method, matched well with the experimentally observed stability. Our study provided evidence that, in addition to steric effects, a small but significant role of electronic effects is governing the configurational stability of such helical quinones. PMID- 17918756 TI - Reinforced self-assembly of hexa-peri-hexabenzocoronenes by hydrogen bonds: from microscopic aggregates to macroscopic fluorescent organogels. AB - Hexa-peri-hexabenzocoronene derivatives (HBCs) that have hydrogen-bonding functionalities (either amido or ureido groups) adjacent to the aromatic cores have been synthesized to study the effects of intracolumnar hydrogen bonds on the self-assembly behavior of HBCs. The hydrogen bonds effectively increased the aggregation tendency of these compounds in solution. In the bulk state, the typical columnar supramolecular arrangement of HBCs was either stabilized substantially (1 a, 1 b, 2 a, and 2 b), or suppressed by dominant hydrogen bonding interactions (3). For some of the compounds (1 a, 2 a, and 2 b), the supramolecular arrangement adopted in the liquid-crystalline state was even retained after annealing, presumably owing to the reinforcement of the pi stacking interactions by the hydrogen bonds. Additionally, the combined effect of the hydrogen bonds and pi-stacking of the aromatic moieties led to the formation of fluorescent organogels, whereby some derivatives were further investigated as novel low molecular-mass organic gelators (LMOGs). PMID- 17918755 TI - Metal-catalysed oxidation processes in thiosemicarbazones: new complexes with the ligand N-{2-([4-N-ethylthiosemicarbazone]methyl)phenyl}-p-toluenesulfonamide. AB - The coordination behaviour of a new thiosemicarbazone Schiff-base building block, N-{2-([4-N-ethylthiosemicarbazone]methyl)phenyl}-p-toluenesulfonamide, H2L1 (1), incorporating a bulky tosyl group, towards Mn II, Fe II, Co II, Ni II, Cu II, Zn II, Cd II, Ag I, Sn II, and Pb II has been investigated by means of an electrochemical preparative procedure. Most metal complexes of L1 have the general formula [M(L1)]2.nX (M=Mn, Fe, Co, Ni, Cu, Cd, Pb; n=0-4, X=H2O or CH3CN), as confirmed by the structure of [Pb(L1)]2 (15), in which the lone pair on lead is stereochemically active. This lead(II) complex shows an intense fluorescence emission with a quantum yield of 0.13. In the case of silver, the complex formed was found to possess a stoichiometry of [Ag2(L1)]2.3H2O. During reactions with manganese and copper metals, interesting catalysed processes have been found to take place, with remarkable consequences regarding the ligand skeleton structure. In synthesising the manganese complex, we obtained an unexpected dithiolate thiosemicarbazone tosyl ligand, H2L2, as a side-product, which has been fully characterised, including by X-ray diffraction analysis. In the case of copper, the solid complex has the formula [CuL1]2, but the crystallised product shows the copper atoms coordinated to a new cyclised thiosemicarbazone ligand, H2L3, as in the structures of the complexes [Cu(L3)]2.CH3CN (8) and [Cu(L3)(H2O)]2.CH3CN.H2O (9). The zinc complex [Zn(L1)]4 (12) displays a particular tetranuclear zeolite-type structure capable of hosting small molecules or ions, presumably through hydrogen bonding. PMID- 17918757 TI - Synthesis of 4-aminotropones from [(sulfinyl or sulfonyl)methyl]-substituted p quinamines. AB - An efficient synthesis of 4-aminotropones has been achieved in excellent yields by simple treatment of 4-amino-4-[(p-tolylsulfinyl)methyl]-2,5-cyclohexadienones (p-quinamines) with NaH. The method allowed regiocontrolled access to 3-methyl, 5 methyl- and 3,5-dimethyl-substituted derivatives starting from p-quinamines with adequate substituents at the cyclohexadienone moiety and/or at the carbon linked to the sulfur function. The p-quinamines in turn were easily accessible from N Boc p-anisidines (Boc=tert-butoxycarbonyl) by electrochemical oxidation in MeOH to quinone imine monoketals, followed by addition of a alpha-lithium sulfinyl carbanion to the imino group, and ketal hydrolysis. Oxidation of the sulfoxide gave the sulfonyl-substituted p-quinamines that, upon basic treatment, behave similarly. The p-quinamine 55 and bis-p-quinamine 56, resulting in the addition of the anion derived from dimethyl sulfone to the p-quinonimine ketal 14, also gave the 4-aminotropone. The mechanism involves the initial formation of a alpha sulfonyl carbanion, which intramolecularly attacks the cyclohexadienone giving a norcaradiene-like enolate intermediate, the evolution of which through a ring expansion process, pushes off a methyl sulfinate anion or SO2. This efficient process fulfils the criteria of atom economy. The introduction of a proline substituent in the nitrogen of the starting p-quinamine allowed the synthesis of an enantiopure 4-aminotropone, the asymmetric Diels-Alder reactions of which with maleimide occurred in a highly endo and pi-facial diastereoselective manner. PMID- 17918758 TI - On the origin of the stereoselectivity in organocatalysed reactions with trimethylsilyl-protected diarylprolinol. AB - The origin of the enantioselectivity in the TMS-protected (TMS=trimethylsilyl) prolinol-catalysed alpha-heteroatom functionalisation of aldehydes has been investigated by using density functional theory calculations. Eight different reaction paths have been considered which are based on four different conformers of the TMS-protected prolinol-enamine intermediate. Optimisation of the enamine structures gave two intermediates with nearly the same energy. These intermediates both have an E configuration at the C==C bond and the double bond is positioned anti or syn, relative to the 2-substituent in the pyrrolidine ring. For the four intermediates, the chiral TMS-protected-diaryl substituent effectively shields one of the faces of the reacting C==C bond in the enamine intermediate. A number of transition states have been calculated for the enantioselective fluorination by N-fluorobenzenesulfonimide (NFSI) and based on the transition-state energies it has been found that the enantioselectivity depends on the orientation of the C==C bond, being anti or syn, relative to the 2 substituent on the pyrrolidine ring, rather than the approach of the electrophilic fluorine to the face of the reacting carbon atom in the enamine which is less shielded relative to the face with the highest shielding. The calculated enantiomeric excess of 96 % ee (ee=enantiomeric excess) for the fluorination reaction corresponds well with the experimentally found enantiomeric excess-97 % ee. The transition state for the alpha-amination reaction with the same type of intermediate has also been calculated by using diethyl azodicarboxylate as the amination reagent. The implication of the intermediate structures on the stereoselection of alpha-functionalisation of aldehydes is discussed. PMID- 17918759 TI - Mu opioid receptor antagonists: recent developments. AB - For thousands of years mu opioid agonists such as morphine have been utilized for their analgesic properties. Today, morphine and related compounds are still used as a first line therapy in the treatment of moderate to severe pain. However, despite the clear benefits of mu agonists in pain management, severe side effects such as dependence and respiratory depression are associated with use of these drugs. To date, there are only two approved mu opioid antagonists for use in the treatment of these adverse effects, that is, naloxone and naltrexone. However, many other clinical and therapeutic areas have been linked to mu opioid receptor antagonism. These include treatment of opioid induced pruritus of the skin, obesity, and Parkinson-induced tardive dyskinesia. Currently there are two compounds, N-methylnaltrexone and alvimopan, under FDA review as possible treatments for opioid induced bowel dysfunction and postoperative ileus. These compounds are of special interest as they are peripherally restricted. This attribute enables treatment of peripheral side effects induced by opioid agonists without reversal of the centrally mediated analgesia of the agonist. In this article we discuss the structural classes of mu opioid antagonists, their potential clinical applications, and review the relevant patents of the last ten years. PMID- 17918760 TI - Betraying the parasite's redox system: diaryl sulfide-based inhibitors of trypanothione reductase: subversive substrates and antitrypanosomal properties. PMID- 17918761 TI - Organometallic ruthenium inhibitors of glutathione-S-transferase P1-1 as anticancer drugs. AB - Ruthenium-arene complexes conjugated to ethacrynic acid were prepared as part of a strategy to develop novel glutathione-S-transferase (GST) inhibitors with alternate modes of activity through the organometallic fragment, ultimately to provide targeted ruthenium-based anticancer drugs. Enzyme kinetics and electrospray mass spectrometry experiments using GST P1-1 and its cysteine modified mutant forms revealed that the complexes are effective enzyme inhibitors, but they also rapidly inactivate the enzyme by covalent binding at Cys 47 and, to a lesser extent, Cys 101. They are highly effective against the GST Pi-positive A2780 and A2780cisR ovarian carcinoma cell lines, are among the most effective ruthenium complexes reported so far, and target ubiquitous GST Pi overexpressed in many cancers. PMID- 17918762 TI - Design and synthesis of isocyanide ligands for catalysis: application to Rh catalyzed hydrosilylation of ketones. AB - New isocyanide ligands with meta-terphenyl backbones were synthesized. 2,6 Bis[3,5-bis(trimethylsilyl)phenyl]-4-methylphenyl isocyanide exhibited the highest rate acceleration in rhodium-catalyzed hydrosilylation among other isocyanide and phosphine ligands tested in this study. 1H NMR spectroscopic studies on the coordination behavior of the new ligands to [Rh(cod)2]BF4 indicated that 2,6-bis[3,5-bis(trimethylsilyl)phenyl]-4-methylphenyl isocyanide exclusively forms the biscoordinated rhodium-isocyanide complex, whereas less sterically demanding isocyanide ligands predominantly form tetracoordinated rhodium-isocyanide complexes. FTIR and 13C NMR spectroscopic studies on the hydrosilylation reaction mixture with the rhodium-isocyanide catalyst showed that the major catalytic species responsible for the hydrosilylation activity is the Rh complex coordinated with the isocyanide ligand. DFT calculations of model compounds revealed the higher affinity of isocyanides for rhodium relative to phosphines. The combined effect of high ligand affinity for the rhodium atom and the bulkiness of the ligand, which facilitates the formation of a catalytically active, monoisocyanide-rhodium species, is proposed to account for the catalytic efficiency of the rhodium-bulky isocyanide system in hydrosilylation. PMID- 17918763 TI - Palladium monomers, dimers, and trimers on the MgO(001) surface viewed individually. PMID- 17918764 TI - A route to three-dimensional structures in a microfluidic device: stop-flow interference lithography. PMID- 17918765 TI - Conformational studies of alanine-rich peptide using CD and FTIR spectroscopy. AB - The circular dichroism (CD) and Fourier transform infrared (FTIR) methods were applied to the conformational studies of alanine-rich peptide Ac-K-[A]11-KGGY-NH2 (where K is lysine, A is alanine, G is glycine and Y is tyrozyne) in water, methanol (MeOH) and trifluoroethanol (TFE). The analysis of CD-spectra of the peptide in water at different concentrations revealed that the secondary structure content depends on the peptide concentration and pH of the solution. The increase of the peptide concentration causes a decrease of alpha-helix content and, simultaneously, an increase of beta-sheet structure, while the unordered structure is the predominant one. Additional elements are discovered in MeOH and TFE but alpha-helix and beta-turns predominate. Moreover, in these solutions the percentage content of the secondary structure does not depend on the temperature. FTIR measurements, carried out at higher peptide concentration (about one order of magnitude) than these CD measurements mentioned above, revealed that in water solution the solid state beta-sheet, and aggregated structures, dominate. However, in TFE the most abundant are alpha-helix and beta turns structures. The thioflavine T assay showed the tendency of the studied peptide for aggregate. PMID- 17918766 TI - Identification of the critical regions in hepatitis B virus preS required for its stability. AB - As a hepatitis B virus (HBV) envelope domain, preS plays significant roles in receptor recognition and viral infection. However, the regions critical for maintaining a stable and functional conformation of preS are still unclear and require further investigation. In order to unravel these regions, serially truncated fragments of preS were constructed and expressed in Escherichia coli. Their solubility, stability, secondary structure, and affinity to polyclonal antibodies and hepatocytes were examined. The results showed that amino acids 31 36 were vital for its stable conformation, and the absence of 10-36 amino acids significantly reduced its binding to polyclonal antibodies as well as hepatocytes. The most stable fragment 1-120 (preS1 + N-terminal 12 amino acids of preS2), perhaps the core of preS, was discovered, which bound to HepG2 cells most tightly. Moreover, the availability of large amounts of well-folded and stable preS1-120 enables us to carry out further structural determination and mechanistic study on HBV infection. PMID- 17918767 TI - Identification, affinity characterisation and biological interactions of lectin like peptide-carbohydrate complexes derived from human TNF-alpha using high resolution mass spectrometry. AB - A cyclic disulfide heptadecapeptide (TIP17ox; 2) derived from the lectin-like 17 amino acid domain of human tumor necrosis factor-alpha [TNF-alpha (100-116)] was synthesised and demonstrated to bind specifically to N,N-diacetylchitobiose, a disaccharide present in many glycan structures of glycoproteins. Although the TIP domain forms a loop structure in the native TNF-alpha protein, we show in this study by high-resolution ESI-FTICR mass spectrometry that a homologous linear heptadecapeptide (TIP17rd; 1) binds with comparable affinity to chitobiose, suggesting that cyclisation is not essential for carbohydrate binding. ESI-FTICR MS was used as an efficient tool for the direct molecular characterisation of TIP peptide-carbohydrate complexes. The specific binding of the TNF-TIP domain to chitobiose and other carbohydrate motifs in glycoproteins may explain the high proteolytic stability of these peptides in biological fluids. A considerably higher proteolytic stability in human plasma was found by mass spectrometric analysis for the cyclic TIP peptide 2, compared to the linear peptide 1. Furthermore, affinity-proteomics studies using immobilised cyclic TIP peptide 2 provided the identification of specific interacting glycoproteins in plasma. PMID- 17918768 TI - Design, synthesis and evaluation of peptide inhibitors of Mycobacterium tuberculosis ribonucleotide reductase. AB - Mycobacterium tuberculosis ribonucleotide reductase (RNR) is a potential target for new antitubercular drugs. Herein we describe the synthesis and evaluation of peptide inhibitors of RNR derived from the C-terminus of the small subunit of M. tuberculosis RNR. An N-terminal truncation, an alanine scan and a novel statistical molecular design (SMD) approach based on the heptapeptide Ac-Glu-Asp Asp-Asp-Trp-Asp-Phe-OH were applied in this study. The alanine scan showed that Trp5 and Phe7 were important for inhibitory potency. A quantitative structure relationship (QSAR) model was developed based on the synthesized peptides which showed that a negative charge in positions 2, 3, and 6 is beneficial for inhibitory potency. Finally, in position 5 the model coefficients indicate that there is room for a larger side chain, as compared to Trp5. PMID- 17918769 TI - Force microscopy imaging of individual protein molecules with sub-pico Newton force sensitivity. AB - The capability of atomic force microscopes (AFM) to generate atomic or nanoscale resolution images of surfaces has deeply transformed the study of materials. However, high resolution imaging of biological systems has proved more difficult than obtaining atomic resolution images of crystalline surfaces. In many cases, the forces exerted by the tip on the molecules (1-10 nN) either displace them laterally or break the noncovalent bonds that hold the biomolecules together. Here, we apply a force microscope concept based on the simultaneous excitation of the first two flexural modes of the cantilever. The coupling of the modes generated by the tip-molecule forces enables imaging under the application of forces ( approximately 35 pN) which are smaller than those needed to break noncovalent bonds. With this instrument we have resolved the intramolecular structure of antibodies in monomer and pentameric forms. Furthermore, the instrument has a force sensitivity of 0.2 pN which enables the identification of compositional changes along the protein fragments. PMID- 17918771 TI - Engineering a circularly permuted GFP scaffold for peptide presentation. AB - The use of peptides as in vivo and in vitro ligand binding agents is hampered by the high flexibility, low stability and lack of intrinsic detection signal of peptide aptamers. Recent attempts to overcome these limitations included the integration of the binding peptide into a stable protein scaffold. In this paper, we present the optimization and testing of a circularly permuted variant of the green fluorescent protein (GFP). We examined the ability of the optimized scaffold to accept peptide insertions at three different regions. The three regions chosen are localized in close spatial proximity to each other and support different conformations of the inserted peptides. In all the three regions peptides with a biased, but still comprehensive, amino acid repertoire could be presented without disturbing the function of the optimized GFP-scaffold. PMID- 17918770 TI - Apathy and depression in Alzheimer's disease are associated with functional deficit and psychotropic prescription. AB - BACKGROUND: Apathy and depression are the most common neuropsychiatric features in Alzheimer's disease (AD). The clinical and functional specific correlates of these syndromes are not well known independently from cognitive deficits and other behavioral disturbances. MATERIAL AND METHODS: Six hundred and eighty-six patients diagnosed with possible or probable AD were included in a prospective multicenter study (REAL-FR). They had an assessment of their cognitive and functional status. Neuropsychiatric symptoms were assessed with the Neuropsychiatric Inventory (NPI) and caregiver's burden was measured with Zarit's Burden Scale. RESULTS: A majority of patients at any stage of the disease presented with one or several behavioral and psychological disturbances. Apathy concerned 43% of patients and, with or without depression, was associated with more pronounced deficits in global cognition, everyday life and instrumental abilities, nutritional status and with a higher burden level. A high level of psychotropic prescription, especially with antidepressant, was observed in patients with apathy. In a multivariate analysis taking into account the cognitive and functional variables of AD, apathy and depression were the only significant predictors of psychotropic prescription. CONCLUSION: Some negative neuropsychiatric symptoms such as apathy and depression have a specific relation with functional and therapeutic outcomes of AD, independently from cognitive status. Further studies are needed to establish if apathy represents a particularly severe phenotype of AD. PMID- 17918772 TI - Continuous drip infusion of low dose cytarabine and etoposide with granulocyte colony-stimulating factor for elderly patients with acute myeloid leukaemia ineligible for intensive chemotherapy. AB - BACKGROUNDS AND OBJECTIVES: The optimal strategy for the management of elderly patients with acute myeloid leukaemia (AML) is still controversial. We previously reported the effectiveness of low dose cytarabine (Ara-C) and etoposide (VP-16) (AV therapy) for those elderly AML patients ineligible for intensive chemotherapy. We initiated the present feasibility study to improve the efficacy by using glanulocyte-colony stimulating factor (G-CSF) with AV therapy (AVG therapy). PATIENTS AND METHODS: The eligibility for enrolment was AML patients according to the World Health Organization (WHO) criteria who were over 60 years of age and who had difficulty in tolerating intensive chemotherapy due to their poor performance status (PS) or some comorbidities. They were given continuous drip infusion of Ara-C (20 mg/body) and VP-16 (50 mg/body) for 7-14 days, and were also simultaneously administered G-CSF (150 microg/m2) once daily. RESULTS: The median age of consecutively enrolled 25 patients was 73 years. Eighteen (72%) patients achieved complete remission (CR). The 1-year overall survival (OS) and the 3-year OS rates were 69% and 22%, respectively. The 1-year disease free survival (DFS) rate in CR patients was 44%. The major regimen related toxicities of grade 3 or 4 were only febrile neutropenia in 15 patients (60%). No regimen related mortality was observed. CONCLUSION: AVG therapy was therefore found to be an effective and well-tolerated regimen for remission induction in elderly AML patients with poor PS or comorbidity. PMID- 17918773 TI - Population-based drug-related anaphylaxis in children and adolescents captured by South Carolina Emergency Room Hospital Discharge Database (SCERHDD) (2000-2002). AB - PURPOSE: Anaphylaxis is a life-threatening condition; drug-related anaphylaxis represents approximately 10% of all cases. We assessed the utility of a statewide emergency department (ED) database for identifying drug-related anaphylaxis in children by developing and validating an algorithm composed of ICD-9-CM codes. METHODS: There were 1 314,760 visits to South Carolina (SC) emergency departments (EDs) for patients <19 years in 2000-2002. We used ICD-9-CM disease or external cause of injury codes (E-codes) that suggested drug-related anaphylaxis or a severe drug-related allergic reaction. We found 50 cases classifiable as probable or possible drug-related anaphylaxis and 13 as drug-related allergic reactions. We used clinical evaluation by two pediatricians as the 'alloyed gold standard'1 for estimating sensitivity, specificity, and positive predictive value (PPV) of our algorithm. RESULTS: ED-treated drug-related anaphylaxis in the SC pediatric population was 1.56/100,000 person-years based on the algorithm and 0.50/100,000 person-years based on clinical evaluation. Assuming the disease codes we used identified all potential anaphylaxis cases in the database, the sensitivity was 1.00 (95%CI: 0.79, 1.00), specificity was 0.28 (95%CI: 0.16, 0.43), and the PPV was 0.32 (0.20, 0.47) for the algorithm. Sensitivity analyses improved the measurement properties of the algorithm. CONCLUSIONS: E-codes were invaluable for developing an anaphylaxis algorithm although the frequently used code of E947.9 was often incorrectly applied. We believe that our algorithm may have over ascertained drug-related anaphylaxis patients seen in an ED, but the clinical evaluation may have under-represented this diagnosis due to limited information on the offending agent in the abstracted ED records. Post-marketing drug surveillance using ED records may be viable if clinicians were to document drug related anaphylaxis in the charts so that billing codes could be assigned properly. PMID- 17918774 TI - Effects of indoor gardening on sleep, agitation, and cognition in dementia patients--a pilot study. AB - OBJECTIVE: A pilot study was performed to examine the efficacy of indoor gardening on sleep, agitation and cognition of dementia patients. METHOD: Twenty three institutionalized dementia patients who had sleep disturbance and/or agitation participated in a 5-week study protocol of 1 week of baseline and 4 weeks of treatment. The study design was a one group repeated measures study. For the first and fifth week of the study period, sleep patterns, agitation, and cognition were evaluated using a sleep diary, Modified Cohen-Mansfield Agitation Inventory and revised Hasegawa Dementia Scale respectively. RESULTS: Significant improvement in wake after sleep onset, nap, nocturnal sleep time, and nocturnal sleep efficiency was identified. On the contrary sleep onset time, wake-up time, total sleep time did not change after indoor gardening. Agitation and cognition score was significantly improved. CONCLUSION: Indoor gardening was found to be effective for sleep, agitation, and cognition of dementia patients. Randomized controlled studies of larger sample size are needed to confirm treatment effect. PMID- 17918776 TI - Strongly coupled versus uncoupled spin response to radio frequency interference effects: application to glutamate and glutamine in spectroscopic imaging. AB - It is well known that comparable radio frequency (RF) wavelengths and human head dimensions at high fields can lead to an inhomogeneous RF field when using standard RF transmission. However, the impact of RF inhomogeneity on potential differences in quantification between coupled and uncoupled spins at longer echo times has not been investigated thoroughly. The consequence of this RF interference on metabolite quantification in spectroscopic imaging at 4.7 T was investigated for the strongly coupled spin systems of glutamate and glutamine at an echo time of 120 ms, and compared with the singlet response of choline. These effects were studied using a single-voxel PRESS sequence (alpha-2alpha-2alpha) with varying flip angle (alpha) from 90 degrees to 65 degrees in simulation, phantom, and in vivo experiments. Phantom metabolite yield decreased to 57% for choline and 27% for glutamate/glutamine in agreement with the simulations. Even a minor reduction from alpha = 85 degrees to 80 degrees produced a large difference between coupled and uncoupled yields, with a reduction of 7% for choline and 17% for glutamate/glutamine. Anecdotal in vivo spectroscopic imaging studies show similar trends, with large differences between choline and glutamate/glutamine yield over a small, 2.2 cm, region. These results demonstrate severe effects on metabolite yield due to RF variation between strongly coupled and uncoupled spin systems at long echo time, which complicates metabolite quantification. PMID- 17918775 TI - The new acyl-CoA cholesterol acyltransferase inhibitor SMP-797 does not interact with statins via OATP1B1 in human cryopreserved hepatocytes and oocytes expressing systems. AB - It is possible that SMP-797 will be administered frequently in combination with statins to hyperlipidemic patients. OATP1B1 is thought to be the major transporter that mediates the hepatic uptake of statins. This study investigated whether SMP-797 interacts with statins via OATP1B1 by two approaches. Firstly, the effects of SMP-797 on OATP1B1-mediated uptake of estrone-3-sulfate (ES) by human hepatocytes and by oocytes expressing OATP1B1 were examined. Since OATP1B1 mediated uptake of ES is known to be biphasic (high- and low-affinity sites), concentrations of [(3)H]ES were set lower than the respective K(m) values. Two representative statins were used to assure that statins share OATP1B1 with [(3)H]ES in this in vitro system. Rosuvastatin inhibited OATP1B1-mediated uptake of [(3)H]ES through both sites and pravastatin inhibited a high-affinity site. The inhibition by the positive control (cyclosporin A) was significant. Thus, it is considered that this system was applicable to examine drug-drug interaction with statins on OATP1B1-mediated uptake. In this condition, no apparent inhibition to each site by SMP-797 was observed up to a concentration 3,000-fold higher than the clinical level. Secondly, the uptake of [(14)C]SMP-797 by oocytes expressing OATP1B1 was examined and the activity was negligible. In conclusion, these data suggest that SMP-797 has little potential to interact with statins on OATP1B1. PMID- 17918779 TI - Characterization of DESI-FTICR mass spectrometry - from ECD to accurate mass tissue analysis. AB - Implementation of desorption electrospray ionization (DESI) technique on a 9.4 T Fourier transform ion cyclotron resonance (FTICR) mass spectrometer is described. Desorption electrospray technique is capable of the direct investigation of natural samples without any need for sample preparation or chromatographic separation. Since the DESI mass spectra of natural samples are very complex owing to the lack of preseparation or cleanup, the ideal mass spectrometric analyzer for these applications is a high-resolution instrument such as FTICR mass spectrometer. DESI was implemented by constructing an electronically controlled source framework comprising six linear moving stages and one rotating stage. A three-dimensional linear stage was used to accommodate samples, while another 3D linear stage equipped with rotating stage was used as a spray mount. A modified electrosonic sprayer was used as a primary electrospray device. DESI-FTICR setup was characterized with regard to geometrical, electrical and flow conditions using deposited peptide samples in range of 1-100 pmol gross deposited amount on glass and polymer surfaces. Optimized conditions enabled the routine acquisition of DESI-MS spectra on the instrument at 130 000 resolution in the broadband mode and with comparable sensitivity to data reported in the literature. Since the main significance of DESI-FTICR MS is the combination of intact tissue analysis, the capabilities of the technique were demonstrated by analyzing murine liver samples. Presence of lysophospholipids in the liver tissue was tentatively associated with the lipid metabolism taking place in liver. DESI-FTICR is also a promising technique in the field of peptide analysis due to capability of top down sequencing using electron capture dissociation. As a proof-of-principle experiment, a small synthetic polypeptide containing 36 amino acids was ionized using DESI and was sequenced in the FTICR by means of ECD (electron capture dissociation) fragmentation. Spectra gave almost full sequence information in agreement with the known amino acid sequence of the species. PMID- 17918777 TI - Impact of field strength and iron oxide nanoparticle concentration on the linearity and diagnostic accuracy of off-resonance imaging. AB - Off-resonance imaging (ORI) techniques are being increasingly used to image iron oxide imaging agents such as monocrystalline iron oxide nanoparticles (MION). However, the diagnostic accuracy, linearity, and field dependence of ORI have not been fully characterized. In this study, the sensitivity, specificity, and linearity of ORI were thus examined as a function of both MION concentration and magnetic field strength (4.7 and 14 T). MION phantoms with and without an air interface as well as MION uptake in a mouse model of healing myocardial infarction were imaged. MION-induced resonance shifts were shown to increase linearly with MION concentration. In contrast, the ORI signal/sensitivity was highly non-linear, initially increasing with MION concentration until T2 became comparable to the TE and decreasing thereafter. The specificity of ORI to distinguish MION-induced resonance shifts from on-resonance water was found to decrease with increasing field because of the increased on-resonance water linewidths (15 Hz at 4.7 T versus 45 Hz at 14 T). Large resonance shifts ( approximately 300 Hz) were observed at air interfaces at 4.7 T, both in vitro and in vivo, and led to poor ORI specificity for MION concentrations less than 150 microg Fe/mL. The in vivo ORI sensitivity was sufficient to detect the accumulation of MION in macrophages infiltrating healing myocardial infarcts, but the specificity was limited by non-specific areas of positive contrast at the air/tissue interfaces of the thoracic wall and the descending aorta. Improved specificity and linearity can, however, be expected at lower fields where decreased on-resonance water linewidths, reduced air-induced resonance shifts, and longer T2 relaxation times are observed. The optimal performance of ORI will thus likely be seen at low fields, with moderate MION concentrations and with sequences containing very short TEs. PMID- 17918780 TI - Disruption of genes encoding pyruvate dehydrogenase kinases leads to retarded growth on acetate and ethanol in Saccharomyces cerevisiae. AB - Two open reading frames, YIL042c (PKP1) and YGL059w, with 25% sequence similarity to human pyruvate dehydrogenase kinases, were shown to have protein kinase activity. Using GFP fusions, it was demonstrated that the proteins localize in discrete submitochondrial regions. Strains with a null mutation in these loci grew poorly on acetate and ethanol as carbon sources. Doubling times increased from ca. 4 h in the wild-type to > 6 h for the mutants. Growth rates of the mutants could be restored to wild-type levels by simultaneous disruption of the PDA1 gene, encoding the E1alpha subunit of the pyruvate dehydrogenase complex. This observation and the pyruvate dehydrogenase activities measured in the mutant strains and the wild-type grown on glucose or acetate suggest that the slow growth phenotype on C2 carbon sources is caused by a futile cycle in which phosphoenolpyruvate is converted back to acetyl coenzyme A. PMID- 17918781 TI - On informative detection bias in screening studies. AB - The problem of estimating the effect of hormone replacement therapy (HRT) on breast cancer has attracted much attention in the causal inference literature. It is known that the HRT effect on diagnosis (observed) is potentially larger than the effect on true cancer status (of interest), and that the standard approach (stratification) to deal with this detection bias is not valid when screening pattern and cancer status share common causes. We propose a sensitivity analysis to deal with detection bias, and demonstrate that the earlier proposed approach of data restriction is not valid. PMID- 17918782 TI - The frequency of anticardiolipin antibodies and genetic mutations associated with hypercoagulability among patients with Wegener's granulomatosis with and without history of a thrombotic event. AB - OBJECTIVE: Venous thrombotic events (VTE), including both deep venous thrombosis and pulmonary emboli, are now recognized as an important complication of Wegener's granulomatosis (WG), but the mechanism(s) of this occurrence is unclear. The frequency of anticardiolipin antibodies (aCL), anti-beta2 glycoprotein antibodies (anti-beta2-GP), and several genetic hypercoagulable factors were examined in a large cohort of patients with WG. METHODS: One hundred eighty patients with active WG had serum and DNA samples collected upon entry into a clinical trial. Of the 180 patients, 29 patients had VTE -- 13 before trial entry, 16 during trial. aCL (IgG, IgM, and IgA) and anti-beta2-GP (IgG and IgM) were evaluated in 176 patients. Factor V Leiden (FVL), the prothrombin gene mutation (G20210A, PGM), and methylenetetrahydrofolate reductase (MTHFR) gene mutation were tested in the 29 patients with thrombotic events, and 36 patients without. RESULTS: aCL occurred with increased frequencies in patients with WG when compared to the general population (1%-5%): 12% had aCL and 3% had anti beta2-GP. There was no difference in the prevalences of aCL or anti-beta2-GP based on clotting status. The prevalence of the genetic hypercoagulable factors examined in patients with WG was comparable to the reported rates in the general population. CONCLUSION: Although the incidence of clinically significant VTE is increased in patients with WG, this increased risk is not explained by increased prevalences of aCL, anti-beta2-GP, FVL, or mutations in PGM or MTHFR. These observations suggest a need to search for new genetic or acquired prothrombotic abnormalities to account for the increased thrombotic event rate in patients with active WG. PMID- 17918783 TI - Estimating the burden of scleroderma disease in Spain. AB - OBJECTIVE: Scleroderma (systemic sclerosis) is a rare disease that results in great suffering. We estimated the burden of disease posed by scleroderma and its relative importance in the health of the Spanish population. METHODS: We estimated scleroderma-based burden of disease following procedures developed for the Global Burden of Disease study to ensure comparability. RESULTS: Despite its low prevalence, scleroderma generated 1732 disability-adjusted life-years (DALY) in Spain in 2001, comprising 562 (32%) years of life lost and 1170 (68%) years lived with disability. Most scleroderma-related DALY (73%) occurred in the population aged 15-54 years. Estimated DALY accounted for 0.59% of other musculoskeletal disorder-related DALY in the European A subregion (countries with low mortality rate in both adults and children in the World Health Organization classification), a significant value in the overall burden of disease. CONCLUSION: The burden of scleroderma in Spain was high, with disability being the major contributing factor. Burden of disease is an important measure in rare diseases, and may be an important indicator to be considered as a health unit in developed countries. PMID- 17918784 TI - In vitro observations of T cell responsiveness to recall antigens during tumor necrosis factor-alpha-blocking therapy in patients with ankylosing spondylitis. AB - OBJECTIVE: Anti-tumor necrosis factor-alpha (TNF-alpha) therapy can induce reactivation of tuberculosis and an increase of other infections in patients with ankylosing spondylitis (AS). This raises the question if an alteration of T cell function can be detected by in vitro analysis to identify patients who might be more at risk of acquiring such infectious diseases. METHODS: We examined peripheral blood from AS patients without history of tuberculosis before and after 10-14 and 24-36 weeks of therapy with adalimumab (n = 8) or infliximab (n = 10). Fresh peripheral blood mononuclear cells were stimulated with cytomegalovirus antigens and with the Mycobacterium tuberculosis antigen purified protein derivative and early secretory antigen target 6. Interferon-gamma production of CD4+ T cells was assessed after in vitro antigen-specific stimulation by intracellular cytokine staining and flow cytometry. RESULTS: There was no significant change, either decrease or increase, of the T cell response to recall antigens during therapy compared to controls without treatment, if the mean values of all patients treated with adalimumab or infliximab were compared at the given timepoints. However, analysis on the individual patient level of such T cell responses revealed 1 adalimumab-treated patient and 2 infliximab treated patients with a clear decrease of T cell response during therapy. Longterm analysis indicated that such a decrease of T cell responsiveness is generally transient and reconstituted at the latest after 52 weeks. CONCLUSION: Some patients treated with adalimumab or infliximab showed a decrease of T cell responsiveness, which seems to be transient. These patients in particular might be at risk for intracellular infections. PMID- 17918785 TI - Male osteoporosis with vertebral fractures? Look for ankylosing spondylitis! A report of 10 cases. AB - OBJECTIVE: Several authors have described the association of ankylosing spondylitis (AS) and osteoporosis. Usually vertebral fractures complicate severe AS. METHODS: We report a series of 10 men in whom benign spondyloarthropathy was discovered during etiological investigation for osteoporosis. Eight patients had B27+ AS and 2 had psoriatic arthritis with axial involvement. The mean number of vertebral fractures was 1.5. No patient had an appendicular fracture. RESULTS: Phosphorus and calcium levels and measurements of 25OHD3, parathyroid hormone, osteocalcin, and serum CTX were in the normal range. The decrease in bone mineral density was greater in the spine (mean T-score at L2-L4 -2.95, mean total hip T score -1.67). CONCLUSION: Osteoporosis with fractures may reveal benign spondyloarthropathy whose clinical expression is sometimes incomplete. Our findings demonstrate that osteoporosis is not always correlated with the severity of AS. PMID- 17918786 TI - Bone edema determined by magnetic resonance imaging reflects severe disease status in patients with early-stage rheumatoid arthritis. AB - OBJECTIVE: To determine the significance of bone edema, detected by magnetic resonance imaging (MRI), in early-stage rheumatoid arthritis (RA). METHODS: We simultaneously examined serologic variables, MRI of wrist sites and finger joints of both hands, clinical disease activity score (DAS), and HLA-DR typing at entry in 80 patients with early-stage RA. RESULTS: The number of bones scored as positive for bone edema correlated with the number of sites scored as positive for MRI synovitis and MRI bone erosion, rate of enhancement (E-rate), and serum C reactive protein (CRP), matrix metalloproteinase 3 (MMP-3), and interleukin 6 (IL 6). Findings for MRI synovitis and MRI bone erosion, E-rate, CRP, MMP-3, IL-6, seropositivity, and titer of anti-cyclic citrullinated peptide antibody (anti-CCP antibody), DAS28-CRP and HLA-DRB1*0405 allele carriership, were significantly higher in the positive versus the negative bone edema group. CONCLUSION: Bone edema based on our scoring system may reflect severe disease status in patients with early-stage RA. However, its clinical value at entry in prognostication of RA should be examined through prospective clinical followup studies. PMID- 17918787 TI - Antiinflammatory mediator lipoxin A4 and its receptor in synovitis of patients with rheumatoid arthritis. AB - OBJECTIVE: To evaluate the role of an antiinflammatory lipid mediator, lipoxin A4 (LXA4), in inflammatory arthritis, we measured the level of LXA4 in synovial fluid and lipoxin A4 receptor (ALX) expression in synovial tissues obtained from patients with rheumatoid arthritis (RA) and osteoarthritis (OA). METHODS: Levels of LXA4 and its analog (15-epi-LXA4) in synovial fluid from 30 patients with RA and 15 patients with OA were measured by a specific ELISA. Reverse transcription polymerase chain reaction (RT-PCR), real-time quantitative PCR, and in situ hybridization were performed to detect mRNA for ALX and 15-LOX, and LXA4 synthetase, in synovial tissues from 20 patients with RA and 10 patients with OA. RESULTS Both LXA4 and 15-epi-LXA4 showed significantly higher levels in RA synovial fluid (10.34 +/- 14.12 ng/ml for LXA4) than OA synovial fluid (0.66 +/- 0.77 ng/ml for LXA4). Logarithmic concentration of LXA4 was significantly correlated with that of leukotriene B4 and prostaglandin E2 in RA and OA synovial fluids. Expressions of ALX and 15-LOX mRNA were stronger in RA synovium than OA synovium. Expression of mRNA for interleukin 13 (IL-13), which induces 15-LOX, was significantly stronger in RA synovium than OA synovium. CONCLUSION: ALX is an important target of LXA4 in synovial tissues of patients with RA. 15-LOX induced by IL-13 might regulate the production of LXA4 to have an antiinflammatory effect against proinflammatory lipid mediators in inflamed joints. These findings could lead to the development of new therapy for inflammatory arthritis such as RA. PMID- 17918789 TI - Radiotherapy fractionation in the beginning of the 21st Century. PMID- 17918788 TI - Improvement and longterm maintenance of quality of life during treatment with adalimumab in severe rheumatoid arthritis. AB - OBJECTIVE: In patients with longstanding severe rheumatoid arthritis (RA) receiving chronic treatment with adalimumab, health related quality of life (HRQOL) was assessed using new instruments [Functional Assessment of Chronic Illness Therapy-Fatigue scale (FACIT-Fatigue) and Health Utilities Index Mark 3 (HUI3)] and a more conventional instrument [Medical Outcomes Study Short Form-36 Health Survey (SF-36)]. METHODS: Different measures for collecting patient reported outcomes were applied simultaneously during the 3-year study period. Sociodemographic and medical history data were assessed at the baseline visit. Clinical examinations (e.g., joint examination and morning stiffness), disease assessments, and HRQOL data were recorded every 8 weeks. For dichotomous and categorical variables, absolute and relative frequencies were calculated. Metric measures were described using mean and standard deviation and/or standard error of the mean. HRQOL data were analyzed using observed cases. RESULTS: All assessed measures (FACIT-Fatigue, HUI3, SF-36) showed a rapid and statistically significant improvement from baseline following initiation of adalimumab therapy. This effect was maintained over the study period for a mean of 1.6 years in all applied measures. HRQOL data from all tested instruments were significantly correlated with each other. CONCLUSION: Chronic therapy with adalimumab improved measures of fatigue and HRQOL in patients with longstanding RA. PMID- 17918790 TI - Immunohistochemical prognostic markers in bladder cancer. AB - The management of superficial bladder cancer is characterized by early recognition of recurrence and the prevention of progression. Biological markers must provide additional information to that provided by multiplicity, size and grade. So, the identification of new prognostic biomarkers for bladder cancer could be used to divide patients into risk groups determining treatment and follow-up schemes. The transformation of a normal cell into a malignant cell is a multistep mechanism, which involves various alterations on the molecular and genetic level. Transitional-cell carcinoma (TCC) is believed to arise through a series of genetic changes. One mechanism of these changes is the mutation of normal genes to oncogenes, which allows cells to escape the regulation of cellular growth control. In addition, tumour growth is regulated by a balance between proliferation, growth arrest and programmed cell death. The control of growth arrest and apoptosis plays key roles in the development of human cancer and in cancer treatment. Tumours present a complex ecosystem in which interactions between tumour cells, extracellular matrix and host cells may lead to reciprocal influences resulting in tumour promotion, invasion and metastasis. Tumour invasion is considered to be a dynamic complex process, which involves a number of steps. During these steps, a lot of critical molecules are involved, the expression of which may offer useful information about the biological behavior of bladder cancer. This review article summarizes the most promising immunohistochemical prognostic factors which have been considered to be involved in the complex and multistep process of bladder carcinogenesis, and in the process of invasion and metastasis. PMID- 17918791 TI - The role of radiotherapy in the management of diabetes insipidus caused by Langerhans cell histiocytosis. AB - Diabetes insipidus (DI) occurs in 5-50% of patients with a diagnosis of Langerhans cell histiocytosis (LCH). DI is the most common manifestation of central nervous system (CNS) involvement and is due to defects in the function of the posterior pituitary. Radiotherapy is recommended as early as possible after the diagnosis of DI, because response rates increase and the need for pitressin therapy may be obviated. The optimal period for radiotherapy seems to be less than one week. Generally, a total radiotherapy dose of 15-20 Gy or less is preferred. Further prospective studies are strongly needed to compare recurrence rates, late effects, disease course, and survival in treated and untreated groups. PMID- 17918792 TI - Accelerated radiotherapy in locally advanced head-neck carcinomas: are concomitant boost and chemotherapy feasible in the routine outpatient-based radiotherapy clinic? AB - PURPOSE: Accelerated radiotherapy and concurrent chemoradiotherapy is an effective treatment modality in locally advanced head and neck carcinomas. In this study, we examined the efficacy and feasibility of concomitant boost radiotherapy and chemotherapy in the routine outpatient- based radiotherapy clinic. PATIENTS AND METHODS: Between January 1993 and December 2000, only 51 out of 127 eligible patients were deemed suitable to receive concomitant boost radiotherapy and/or chemotherapy. Their median age was 60 years (range 17-83 years). The histological diagnosis was squamous-cell carcinoma in 38 (75%) patients, undifferentiated nasopharyngeal carcinomas (WHO type III) in 10 (20%) patients and other histology in 3 (5%) patients. The concomitant boost regimen consisted of 70 Gy in 6 weeks (1.8 Gy/fraction/day, 5 days/week, to the clinical target volume (CTV), and 1.6 Gy/fraction/day to the gross tumor volume (GTV) as a second-daily treatment for the last 2 weeks). The concomitant chemotherapy regimen consisted of cisplatin 100 mg/m(2) given every 3 weeks for 3 courses, and the neoadjuvant regimen of cisplatin 100 mg/m(2) plus epirubicin 100mg/m(2), every 3 weeks for 3 courses. RESULTS: Only 55% of patients completed the treatment exactly as planned, with 82% completing treatment within acceptable limits. The median radiotherapy duration was 45 days (40-95 days). In univariate analysis, patients with better performance status (p=0.002) or nasopharyngeal carcinomas (p=0.043) had a significantly better compliance to treatment. In multivariate analysis only nasopharyngeal site was a significant predictor of compliance (p=0.019). The maximum acute reaction was grade 3 mucositis in 24 (49%) patients. No grade 4 acute or late reactions were seen. Complete and overall response rates were 51% and 75.5%, respectively. Patients with nasopharyngeal cancer and good treatment compliance had a better response rate (p=0.009 and 0.01, respectively). The median follow-up period of surviving patients was 28 months (range 6-58 months). The 3-year progression-free survival rate was 39%. CONCLUSION: In the routine outpatient-based setting we found that this intensive treatment schedule can only be given to a limited and highly selected group of patients. PMID- 17918793 TI - Diagnosis and treatment of primary non-Hodgkin's lymphoma of the parotid gland: a retrospective study - Experience at the Northern Israel Oncology Center (1977 1999). AB - PURPOSE: The treatment and outcome of primary parotid gland non-Hodgkin's lymphoma (PGL) has rarely been described. This retrospective study documents the clinicopathologic features and treatment results in this relatively rare entity. PATIENTS AND METHODS: This study was conducted on 11 patients diagnosed and treated for primary PGL over a period of 22 years. RESULTS: Of the 4 male and 7 female patients, only one presented with the classic pattern of Sjogren's syndrome (SS) simultaneous with PGL, and only 4 patients demonstrated a low-grade Maltoma type. None of the patients had evidence of disease at the end of the primary treatment; 4 patients are alive and well from 6 months to 10 years after the end of treatment. Four patients relapsed and died due to therapy-resistant disease and 3 patients died of nonmalignant causes while in complete remission. CONCLUSION: The majority of patients with primary non- Hodgkin's lymphoma of the parotid gland present with early- stage disease. Accurate staging is mandatory. Low-grade, localized PGL can be treated successfully with primary radiotherapy alone. The aggressive type of PGL should be treated with combined chemoradiotherapy-based regimens. PMID- 17918794 TI - Completion pneumonectomy for lung cancer. AB - PURPOSE: Completion pneumonectomy is a trully challenging operation associated with increased mortality and morbidity. The aim of this study was to present a series of 18 patients who underwent completion pneumonectomy for lung cancer during a 15-year period and to evaluate the postoperative outcomes and long-term results. PATIENTS AND METHODS: Between January 1985 and December 2000,18 patients underwent completion pneumonectomy for lung cancer; 10 for local recurrence ,6 for second primary lung tumor and 2 for lung tumors in patients who had previously been operated on for benign disease. RESULTS: No intraoperative deaths occurred. Postoperative mortality and morbidity were 11.11% and 33.33%, respectively. The median operational time was 212.7 minutes. The mean blood loss during the procedure was 1.042,5 ml. The complication rate was 33.33%. The 5-year survival was 18.75% for all patients. The 5-year survival was 25% for the local recurrence group and 50% for the primary lung cancer group. The 5-year survival of the patients in the second primary tumor group has not been reached yet. CONCLUSION: Completion pneumonectomy can be performed with an acceptable operative mortality rate and offers a second chance for cure to patients with lung cancer. Although complications are common ,they can successfully be managed with proper understanding of them. PMID- 17918795 TI - Organ-sparing treatment of invasive transitional cell bladder carcinoma. AB - PURPOSE: To assess the effectiveness of two approaches of organ-sparing treatment in patients with invasive transitional cell bladder carcinoma. PATIENTS AND METHODS: During the period from June 1996 to June 2000, 33 patients with invasive transitional cell carcinoma (T2-4) of the bladder were treated. Patients were divided into two groups. Group A included 17 patients treated with CMV systemic chemotherapy (methotrexate 30 mg/m(2) and vinblastine 3 mg/m(2), day 1; and cisplatin 70 mg/ m(2), day 2) repeated every 3 weeks for 3 courses, combined with intravesical BCG. Complete responders (CR) received maintenance intravesical BCG, while partial responders (PR) were subjected to transurethral bladder resection (TURB) or partial bladder resection. Group B included 16 patients treated with maximal TURB followed by 2 CMV courses and radiotherapy. RESULTS: Group A patients have been followed-up for a period of 7 to 48 months (median 21.6 months). After completing the 2nd chemotherapy course, 2 patients refused further treatment and were excluded from the group. On completing chemoimmunotherapy 11 (73%) patients showed objective response (CR+PR) and preservation of the bladder was achieved. Four (27%) patients were treatment failures. Group B patients have been followed for a period of 9 to 47 months (median 27.5 months). On completing treatment 12 of 16 (75%) patients showed an objective response with preservation of the bladder. Treatment failure was diagnosed in 4 (25%) patients. The 2 groups differed significantly in terms of side-effects, which were more pronounced in Group B patients. Of the late complications in Group B a serious problem was the development of severe radiation-induced fibrosis leading to microcystis and a cystectomy was indispensable in one of the patients. CONCLUSION: The applied 2 approaches of combined organ-sparing treatment of invasive bladder carcinoma seem equally effective. Future randomized studies are needed to define reliable criteria for patient selection. PMID- 17918796 TI - Radical hysterectomy for stage IB1 cervical carcinoma: lymph node metastasis as a prognostic factor. AB - PURPOSE: Lymph node metastasis is one of the most important factors influencing prognosis and further therapy in patients with cervical carcinoma. The aim of this study was to confirm the impact of nodal metastasis od disease-free interval in women with stage IB1 cancer of the uterine cervix. PATIENTS AND METHODS: From June 1986 to December 1999 269 patients with stage IB1 cervical carcinoma were operated on. Two hundred thirty-six (87.84%) patients had class III and 33 (12.16%) class II radical hysterectomy, according to Piver's classification. RESULTS: The median number of the removed lymph nodes was 21. Positive lymph nodes were found in 71 (25.28%) patients. All patients with positive lymph nodes received postoperative adjuvant external beam radiotherapy. Patients with bulky nodal disease received also chemotherapy.Overall 5-year disease-free interval in 212 patients was 80%. Five-year disease-free interval for patients without lymph node metastasis was 91%, while it was 40% in those with lymph node metastasis (p < 0.0001). CONCLUSION: Surgical staging of cervical cancer, which includes pelvic and para-aortic lymphadenectomy,together with pathological data, can provide potentially useful information for the radiation oncologist and precise analysis of survival and prognostic risk factors. PMID- 17918797 TI - Distribution of high-risk HPV types in Yugoslav women with cervical neoplasia. AB - PURPOSE: The purpose of this study was to determine the frequency of high-risk types of human papillomavirus (HPV) infection, namely type 16, 18, 31 and 33, among Yugoslav women diagnosed with different grades of squamous intraepithelial lesion (SIL), as well as to investigate the relationship between HPV infection and age, parity, age at first intercourse, number of sexual partners and residence of the patients, all of which are considered risk-factors. PATIENTS AND METHODS: DNA was isolated from cervical swabs of 72 women using phenol/chloroform/isoamylalcohol extraction. Detection of HPV DNA in patients' genomic DNA was performed using the polymerase chain reaction (PCR) method with type-specific primer pairs, and amplification products were analyzed using 2% agarose and 10% polyacrylamide gel electrophoresis. RESULTS: Thirty out of 72 (41.7%) patients with cervical intraepithelial neoplasia (CIN) were HPV-positive and 8 of them were double positives. HPV31 was the most frequent high-risk HPV type in this group of patients (13.9%). Eighty percent of the high-grade SIL (HSIL) patients were HPV-positive and 38.8% of the low-grade SIL (LSIL) patients were HPV-positive. Compared to HPV-negative women, the HPV-positive ones were younger, had started sexual activity earlier, and overall had more sexual partners. CONCLUSION: Our study showed that oncogenic HPV types are responsible for the transition of LSIL to HSIL, and for its further progression to an invasive carcinoma of the cervix. Thus, HPV typing should become a widely used method for identifying women with increased risk for developing HSIL and invasive cervical cancer. We also concluded that sexual behavior is connected with the frequency of HPV infection. Henceforth, introduction of prophylactic measures could reduce the incidence of HPV infected women in our country. PMID- 17918798 TI - Correlation of imprint cytology and histopathologic findings in bone tumors. AB - PURPOSE: The aim of this study was to show that imprint cytology is a valuable method in the evaluation of bone tumors. MATERIALS AND METHODS: Fifty-six surgical specimens of bone tumors imprints were air-dried and stained by the May Grunwald-Giemsa method. A cytologist gave a detailed description of the smears, concluded whether the lesion was benign or malignant and suggested a final diagnosis. Results were compared with the corresponding histopathologic diagnoses. RESULTS: In 2 cases the material was too scarce for cytologic diagnosis. Out of 19 cases with primary malignant bone tumors the cytologist recognized malignancy in 18 of them and in 10 suggested the final diagnosis. In 14 patients with metastases (11 with squamous-cell carcinoma, 1 with anaplastic carcinoma, 2 with malignant melanoma) the cytologic and histopathologic diagnoses correlated in all cases. In 21 patients with benign bone lesions, the cytologist defined as benign 19 of them and in one case had suspicion of fibrosarcoma. In one patient with a histopathologic diagnosis of bone necrosis, groups of clearly malignant cells were found in the imprint. After revision of the case, the cytologist confirmed the diagnosis of malignancy. CONCLUSION: Cytological analysis of the imprints of bone lesions is valuable for an early orientation of the clinicians. It is very important, since the final histopathologic diagnosis needs time because of the decalcification procedure. PMID- 17918799 TI - Development of a national protocol for a high activity (192)Ir brachytherapy source calibration. AB - PURPOSE: Determination of the air kinetic energy released in material (KERMA) rate (commonly named source strength) or a brachytherapy source activity represents one of the first steps in the acceptance, as well as in the periodic Quality Assurance (QA) testing procedures. In this paper, calibration of an (192)Ir high-dose rate (HDR) brachytherapy source is presented. MATERIALS AND METHODS: An (192)Ir HDR brachytherapy source calibration was carried out, with two independent procedures - calibration in water and in free air. The dosimeter used consisted of a thimble ionization chamber (NE 2571) and an electrometer (NE 2670). The reference air KERMA rate was calculated according to the International Atomic Energy Agency (IAEA) and the International Commission for Radiation Units and Measurements (ICRU) protocols. All physical quantities and correction factors used were determined, calculated, and re-checked for particular chamber and source type. RESULTS: In water, the reference air KERMA rate, determined by the IAEA protocol differed by +1.0% from the manufacturer's declared value and was measured with a relative error of 1%. The reference air KERMA rate, calculated by the ICRU protocol differed by -0.4% from the manufacturer's declared value and was measured with a relative error of 1.1%. In air, the value of the reference air KERMA rate, calculated by the IAEA protocol, differed by +1.6% from the manufacturer's declared value and was measured with a relative error of 1%. The value of the reference air KERMA rate for the same source, calculated by the ICRU protocol, differed by +1.1% from the manufacturer's declared value and was measured with a relative error of 0.9%. The results showed that application of the IAEA protocol gave higher values than the application of the ICRU protocol. It stands for both calibration methods. The difference between calibration protocols was not higher than 2% (in the water 1.4% and in the air 0.5%). Finally, calibration results showed that the difference between the calibration methods (in water and in air), for both calibration protocols, was not higher than 2%. For the IAEA protocol the difference was -0.6% and for the ICRU protocol it was -1.5%. We believe that benefits can be derived from the simultaneous application of both methods. CONCLUSION: A proposal of a national protocol represents an attempt to formulate a unique protocol for brachytherapy sources calibration in Serbia. The comparison of "in water" and "in free air" calibration has shown systematic differences of in-source strength of about 2%, respectively. It would be worth suggesting to radiotherapy centres to adopt the most optimal calibration method, based on well-known international protocols. PMID- 17918800 TI - Successful management of extracranial metastases from cerebral high-grade oligodendroglioma: report of a case with literature review. AB - The prognosis of high-grade oligodendroglioma has been reported poor with an average survival of approximately 17 months. Local recurrence after surgery is seen in about 50% of the patients, but distant extracranial metastases have been described rarely. Frequent extracranial metastases have been reported in the lungs, pleura, lymph nodes, bones, liver, adrenal glands, kidneys and other sites. We present a case of 16-year-old girl with high-grade oligodendroglioma who developed bilateral pulmonary metastases 19 months following complete resection of the primary brain tumor. She did not respond to first-line chemotherapy; however, the patient achieved an almost complete response with cisplatin (CDDP) and etoposide (VP-16) salvage chemotherapy. She remains well and asymptomatic 18 months after the end of chemotherapy. We herein review the literature and discuss all the possible mechanisms for extracranial dissemination of primary brain tumours. PMID- 17918801 TI - Primary Sjogren's syndrome complicated with development of non-Hodgkin's lymphoma - a case report and literature review. AB - Primary Sjogren's syndrome (PSS) is an autoimmune disease of the exocrine glands that is presented with progressive ocular and oral dryness, parotid enlargement and often with systemic (extraglandular) manifestations. In patients with PSS the risk of development of non-Hodgkin's lymphoma (NHL) is 44-fold compared to healthy population. The risk is associated with certain characteristics of the disease's course: recurrent changes in the parotid glands, lymph node enlargement, skin vasculitis, peripheral neuropathy, anemia and lymphopenia. According to the morphologic and phenotypic characteristics, B-cell low-grade lymphomas prevail. This communication presents a 65-year-old woman with PSS who developed a follicular type B-cell NHL 21 years after the autoimmune disease had been diagnosed. The analysis of our case and the literature review summarize the characteristics of the course of PSS, histologic variants, and evolution and prognosis of NHL in this kind of patients. PMID- 17918802 TI - An unusual case of fat necrosis of the breast. AB - A 56-year-old woman receiving Depixol injections for schizophrenia presented with two separate episodes of breast fat necrosis with no identifiable cause. It is postulated that the fat necrosis could be as of a result of the Depixol injections, where this drug is taken up into the breast fat and subsequently rapidly hydrolysed resulting in fat necrosis. PMID- 17918803 TI - Endoscopic diagnosis of ureteral obstruction secondary to metastatic breast carcinoma. AB - Metastasis to the ureter is a very rare complication in patients with breast cancer and bone metastases. Retroperitoneal disease appears to be an uncommon manifestation of long-standing breast cancer which is usually hormonally dependent. In most published cases, the diagnosis is only made at postmortem examination. This report presents an uncommon case of ureteral obstruction secondary to breast cancer diagnosed with ureteroscopy. PMID- 17918804 TI - Incisional metastasis after cervical mediastinoscopy: a case report. AB - The risk of iatrogenic tumor seeding from mediastinoscopy is low. The etiology of this complication remains unclear. We present the case of a patient with this condition, discuss the cause and management, and review the literature. PMID- 17918805 TI - Milestones in the study of bladder tumors and the contribution of Sir Henry Thompson (1820-1904). AB - We tried through this paper to present briefly the history of the study of bladder tumors and to underline the contribution of Sir Henry Thompson - outstanding British urologist - on that field. PMID- 17918806 TI - First-principles calculation of local atomic polarizabilities. AB - Common methods of determining atomic polarizabilities suffer from the inclusion of nonlocal effects such as charge polarization. A new method is described for determining fully ab initio atomic polarizabilities based on calculating the response of atomic multipoles to the local electrostatic potential. The localized atomic polarizabilities are then used to calculate induction energies that are compared to ab initio induction energies to test their usefulness in practical applications. These polarizabilities are shown to be an improvement over the corresponding molecular polarizabilities, in terms of both absolute accuracy and the convergence of the multipolar induction series. The transferability of localized polarizabilities for the alkane series is also discussed. PMID- 17918808 TI - Ultrafast photoinduced dynamics of pigment yellow 101: fluorescence, excited state intramolecular proton transfer, and isomerization. AB - The ultrafast excited-state dynamics of the fluorescent pigment yellow 101 (P.Y.101) and the closely related 1,1'-naphthalazine, a nonfluorescent derivative that lacks the OH groups at the naphthyl rings, are studied combining femtosecond spectroscopy and high-level quantum chemical calculations. The observed ultrafast dynamics and the spectral signature of photoexcited 1,1'-naphthalzine can be consistently explained with a previously proposed mechanism, suggesting fluorescence quenching via an optically forbidden npi* state. In contrast, for a description of the excited-state dynamics of P.Y.101, the expected simple absorption/fluorescence model is not adequate. Instead, besides fluorescence as the main decay channel of the excited-state population, ultrafast excited-state intramolecular proton transfer (ESIPT) and isomerization processes have to be considered for a complete understanding of the complex subnanosecond dynamics. Combining experiment and theory, the following kinetic model is derived: upon photoexcitation a major part of the excited-state population decays via fluorescence from an enol-type isomer of P.Y.101, while a small part of the population undergoes ESIPT and fluoresces from a keto-type form. Furthermore, arguments are given that, to a minor extent, also trans-cis isomerization of the keto form takes place on the S1 surface leading probably to a long-lived cis-keto form in the ground state. The remarkable photostability of this organic pigment is thus achieved by the interplay of different ultrafast nondestructive decay channels. PMID- 17918809 TI - Thermochemistry of 2- and 3-acetylthiophenes: calorimetric and computational study. AB - The relative stabilities of 2- and 3-acetylthiophenes have been evaluated by experimental thermochemistry and the results compared to high-level ab initio calculations. The enthalpies of combustion, vaporization, and sublimation were measured by rotating-bomb combustion calorimetry, Calvet microcalorimetry, correlation gas chromatography, and Knudsen effusion techniques and the gas-phase enthalpies of formation, at T = 298.15 K, were determined. Standard ab initio molecular orbital calculations at the G2 and G3 levels were performed, and a theoretical study on the molecular and electronic structures of the compounds studied has been conducted. Calculated enthalpies of formation using atomization and isodesmic reactions are compared with the experimental data. Experimental and theoretical results show that 2-acetylthiophene is thermodynamically more stable than the 3-isomer. A comparison of the substituent effect of the acetyl group in benzene and thiophene rings has been carried out. PMID- 17918807 TI - Experimental and DFT studies: novel structural modifications greatly enhance the solvent sensitivity of live cell imaging dyes. AB - Structural modifications of previously reported merocyanine dyes (Toutchkine, A.; Kraynov, V.; Hahn, K. J. Am. Chem. Soc. 2003, 125, 4132-4145) were found to greatly enhance the solvent dependence of their absorbance and fluorescence emission maxima. Density functional theory (DFT) calculations have been performed to understand the differences in optical properties between the new and previously synthesized dyes. Absorption and emission energies were calculated for several new dyes using DFT vertical self-consistent reaction field (VSCRF) methods. Geometries of ground and excited states were optimized with a conductor like screening model (COSMO) and self-consistent field (SCF) methods. The new dyes have enhanced zwitterionic character in the ground state and much lower polarity in the excited state, as shown by the DFT-VSCRF calculations. Consistently, the position of the absorption bands are strongly blue-shifted in more polar solvent (methanol compared to benzene), as predicted by the DFT spectral calculations. Inclusion of explicit H-bonding solvent molecules within the quantum model further enhances the predicted shifts and is consistent with the observed spectral broadening. Smaller but significant spectral shifts in polar versus nonpolar solvent are predicted and observed for emission bands. The new dyes show large fluorescence quantum yields in polar hydrogen-bonding solvents; qualitatively, the longest bonds along the conjugated chain at the excited S1 state minimum are shorter in the more polar solvent, inhibiting photoisomerization. The loss of photostability of the dyes is a consequence of the reaction with and electron transfer to singlet oxygen, starting oxidative dye cleavage. The calculated vertical ionization potentials of three dyes I-SO, AI SO(4), and AI-BA(4) in benzene and methanol are consistent with their relative photobleaching rates; the charge distributions along the conjugated chains for the three dyes are similarly predictive of higher reaction rates for AI-SO(4) and AI-BA(4) than for I-SO. Time-dependent DFT calculations were also performed on AI BA(4); these were less accurate than the VSCRF method in predicting the absorption energy shift from benzene to methanol. PMID- 17918810 TI - Ab initio investigation of the complexes between bromobenzene and several electron donors: some insights into the magnitude and nature of halogen bonding interactions. AB - Halogen bonding, a specific intermolecular noncovalent interaction, plays crucial roles in fields as diverse as molecular recognition, crystal engineering, and biological systems. This paper presents an ab initio investigation of a series of dimeric complexes formed between bromobenzene and several electron donors. Such small model systems are selected to mimic halogen bonding interactions found within crystal structures as well as within biological molecules. In all cases, the intermolecular distances are shown to be equal to or below sums of van der Waals radii of the atoms involved. Halogen bonding energies, calculated at the MP2/aug-cc-pVDZ level, span over a wide range, from -1.52 to -15.53 kcal/mol. The interactions become comparable to, or even prevail over, classical hydrogen bonding. For charge-assisted halogen bonds, calculations have shown that the strength decreases in the order OH- > F- > HCO2- > Cl- > Br-, while for neutral systems, their relative strengths attenuate in the order H2CS > H2CO > NH3 > H2S > H2O. These results agree with those of the quantum theory of atoms in molecules (QTAIM) since bond critical points (BCPs) are identified for these halogen bonds. The QTAIM analysis also suggests that strong halogen bonds are more covalent in nature, while weak ones are mostly electrostatic interactions. The electron densities at the BCPs are recommended as a good measure of the halogen bond strength. Finally, natural bond orbital (NBO) analysis has been applied to gain more insights into the origin of halogen bonding interactions. PMID- 17918811 TI - Aggregate of alkoxy-bridged Re(I)-rectangles as a probe for photoluminescence quenching. AB - Alkoxy-bridged rhenium(I) rectangles [{(CO)(3)Re(mu-OR)(2)Re(CO)(3)}(2)(mu bpy)(2)] (1, R = C(4)H(9); 2, R = C(8)H(17); 3, R = C(12)H(25); bpy = 4,4' bipyridine) comprising long alkyl chains form optically transparent aggregates and exhibit luminescence enhancement in the presence of water. The aggregation of Re(I)-rectangle was followed using a light-scattering technique. Presumably, the enhanced luminescence efficiency resulted from restriction of torsional molecular motion in the aggregates. In addition, the rate of bimolecular quenching of Re(I) aggregates in the triplet excited state by various electron donors (amines) and acceptors (quinones) was efficient. These results indicate that the excited state of aggregated Re(I) surfactants with an electron acceptor and donor facilitate the electron-transfer quenching process after they became preassociated inside the Re(I)-aggregated species. These synthesized compounds may be useful fluorescent materials in optoelectronic applications. PMID- 17918812 TI - Spectroscopic probes of the quasi-liquid layer on ice. AB - Raman spectra of the water OH-stretch region were acquired at air-ice and air water interfaces at a glancing angle, which allowed observation of surface characteristics. The shapes of the OH-stretch bands indicate that the environment at the air-ice interface is different from that at the air-water interface and from that seen in bulk water. Water spectra measured at the surface of dodecane under low relative humidity indicate that this method is sensitive to fewer than 50 monolayers of water. Changes in the local environment of the surfacial water molecules may be induced by the presence of different solute species, giving rise to changes in the shape of the band. Dissolved sodium chloride disrupts hydrogen bonding in liquid water and has the same effect at the air-ice interface. However, when either HCl or HNO(3) is adsorbed from the gas phase onto an ice surface, the opposite effect is seen: Their presence appears to increase the extent of hydrogen bonding at the ice surface. At the same time, shifts in the laser-induced fluorescence spectra of acridine, a fluorescent pH-probe present at the air-ice interface, indicate that dissociation of acids occurs there. These observations suggest that the formation of hydronium ions at the air-ice interface enhances the hydrogen bonding of surfacial water molecules. PMID- 17918813 TI - Relationship between the critical points found by the electron localization function and atoms in molecules approaches in adducts with hydrogen bonds. AB - In this work, 11 adducts with hydrogen bonds were studied by using the B3LYP exchange-correlation functional of the Kohn-Sham approach and the Moller-Plesset second-order perturbation theory MP2. With both approaches, the geometry of each adduct was optimized with the aug-cc-pVTZ basis set. The binding energies of the considered systems, found by the MP2 method, range from 1.2 to 8.3 kcal/mol. By using the atoms in molecules (AIM) analysis and the electron localization function (ELF) we found that the critical points positions characteristic of hydrogen bonds obtained by AIM and ELF are very similar each other. Besides, we found a linear correlation between the critical points positions found by AIM and those obtained by ELF with the B3LYP method and also with the MP2 method. The slope of such a linear relationship was close to 1 and the y-intercept close to 0. PMID- 17918814 TI - Spectroscopic signatures of halogens in clathrate hydrate cages. 2. Iodine. AB - UV-vis and Raman spectroscopy were used to study iodine molecules trapped in sII clathrate hydrate structures stabilized by THF, CH(2)Cl(2), or CHCl(3). The spectra show that the environment for iodine inside the water cage is significantly less perturbed than either in aqueous solution or in amorphous water-ice. The resonance Raman progression of I(2) in THF clathrate hydrate can be observed up to v = 6 when excited at 532 nm. The extracted vibrational frequency omega e = 214 +/- 1 cm(-1) is the same as that of the free molecule to within experimental error. At the same time, the UV-vis absorption spectrum of I(2) in the sII hydrate exhibits a relatively large, 1440 cm(-1), blue-shift. This is mainly ascribed to the differential solvation of the I(2) electronic states. We conclude that iodine in sII hydrate resides in a 5(12)6(4) cavity, in which the ground-state I(2) potential is not significantly perturbed by the hydrate lattice. In contrast, in water and in ice, the valence absorption band of I(2) is dramatically broadened and blue-shifted by 3000 cm(-1), and the resonance Raman scattering is effectively quenched. These observations are shown to be consistent with a strong interaction between water molecule and iodine through the lone pair of electrons on water as in the case of bromine in the same media. The results presented here, and the stability of other halogen hydrates, were used to test the predictions of simple models and force-field calculations of the host cage-guest association energy. PMID- 17918815 TI - Theoretical study of the dissociation of nitric acid at a model aqueous surface. AB - The issue of acid dissociation of nitric acid at an aqueous surface is relevant in various portions of the atmosphere in connection with ozone depletion. This proton-transfer reaction is studied here via electronic structure calculations at the HF/SBK+(d) level of theory on the HNO(3) x (H(2)O)(3) model reaction system embedded in clusters comprising 33, 40, 45, and 50 classical, polarizable waters with an increasing degree of solvation of the nitrate group. Free energy estimates for all the cases examined favor undissociated, molecular nitric acid over the 0-300 K temperature range, including that relevant for the upper troposphere, where it is connected to the issue of the mechanism of nitric acid uptake by water ice aerosols. The presence of molecular HNO(3) at 300 K at the surface is further supported by vibrational band assignments in good agreement with a very recent surface-sensitive vibrational spectroscopy study of diluted HNO(3)/H(2)O solutions. PMID- 17918816 TI - A ULFC method for d(4)(D(2d)) ions and a study of the spin singlets contributions to zero-field splitting of Cr(2+) ions in zinc sulfide crystals. AB - A simple theoretical method is shown to yield a detailed explanation of numerous EPR parameters for a d4 configuration ion in tetragonal ligand field. Using the unified ligand-field-coupling (ULFC) scheme, the formulas relating the microscopic spin Hamiltonian parameters with the crystal structure parameters are derived. On the basis of the theoretical formulas, the 210 x 210 complete energy matrices including all the spin states are constructed within a strong field representation. By diagonalizing the complete energy matrices, the local lattice structure and Jahn-Teller energy of Cr(2+) ions in ZnS:Cr(2+) system have been investigated. It is found that the theoretical results are in good agreement with the experimental values. Moreover, the contributions of the spin singlets to the zero-field splitting (ZFS) parameters of Cr(2+) ions in ZnS crystals are investigated for the first time. The results indicate that the spin singlets contributions to ZFS parameter b(0)(4) is negligible, but the contributions to ZFS parameters b(0)(4) and b(4)(4) cannot be neglected. PMID- 17918817 TI - Photoelectron detachment and solvated electron dynamics of the cobalt(III) and iron(III) oxalato complexes. AB - The photochemical redox reaction kinetics of [M(III)(C(2)O(4))(3)](3-) [M = Co, Fe] complexes have been reexamined and studied by time-resolved spectroscopy. The redox mechanisms of the two systems, Co and Fe, were found to be similar to each other, and solvated electrons were observed immediately after 266/267 nm photoexcitation. A reaction mechanism is proposed that involves photoelectron detachment as a primary process. The charge-transfer bands for both complexes, which had been attributed to ligand to metal charge-transfer (LMCT) transitions previously, are reassigned to charge transfer to solvent (CTTS) transitions. PMID- 17918818 TI - Ozonolysis of mixed oleic acid/n-docosane particles: the roles of phase, morphology, and metastable states. AB - The reaction kinetics of ozone with oleic acid (OA) in submicron particles containing n-docosane has been studied using aerosol CIMS (chemical ionization mass spectrometry) to monitor changes in particle composition. Internally mixed particles with X(OA) > 0.72 were found to exist as supercooled droplets when cooled to room temperature. Partial reaction of the oleic acid was seen to completely inhibit further reaction and was attributed to the formation of a metastable solid rotator phase of the n-docosane at the surface. This reaction induced phase change is believed to prevent further reaction by slowing ozone diffusion into the particle. When these particles were cooled to 0 degrees C before reaction, they reacted to a further extent and did not demonstrate such an inhibition. This shift in reactivity upon cooling is attributed to the formation of the thermodynamically stable form of n-docosane, the triclinic solid. This transition was accompanied by an increase in the n-docosane density, which led to the development of "cracks" through which ozone can diffuse into the particle. The aerosol with X(OA) < 0.72 consisted of an external mixture of particles containing n-docosane in either the rotator or the triclinic solid phase because of the stochastic nature of the rotator --> triclinic transition. The reactivity of the oleic acid was seen to increase with increasing n-docosane content as a larger fraction of the particles underwent the rotator --> triclinic transition and therefore contained cracks at the surface. These findings demonstrate the importance of transient, metastable phases in determining particle morphology and how such morphological changes can influence rates of reactions in organic aerosols. PMID- 17918819 TI - Synthesis and structure-activity relationship for a novel class of potent and selective carbamate-based inhibitors of hormone selective lipase with acute in vivo antilipolytic effects. AB - Hormone-sensitive lipase (HSL) is an intracellular enzyme that has a central role in the regulation of fatty acid metabolism. The enzyme, therefore, is a potentially interesting pharmacological target for the treatment of insulin resistance and dyslipidemic disorders. Based on a high throughput screening, a carbamate based HSL inhibitor was identified and optimized into the selective HSL inhibitors 4-hydroxymethyl-piperidine-1-carboxylic acid 4-(5 trifluoromethylpyridin-2-yloxy)-phenyl ester (13f) and 4-hydroxy-piperidine-1 carboxylic acid 4-(5-trifluoromethylpyridin-2-yloxy)-phenyl ester (13g), with IC50 values of 110 and 500 nM, respectively. Both inhibitors were active in acute antilipolytic experiments in vivo and none of the inhibitors inhibited the cytochrome P450 (CYP) isoforms 2D6, 3A4, and 1A2. PMID- 17918820 TI - Target specific virtual screening: optimization of an estrogen receptor screening platform. AB - In this work, we introduce a four-step scoring and filtering procedure, furnishing target specific virtual screening (TS-VS), which serves to minimize false positives resulting from conformational artifacts of the docking process and is optimized to converge on novel chemotypes of estrogen receptor alpha (ERalpha). As a proof of concept, VS of a commercial compound database was undertaken (SPECs database release: Aug 2005, 202 054 compounds in total), resulting in the identification of both previously known and novel putative ER scaffolds. Application of distance constraints within TS-VS allowed facile identification of three novel active ligands with ERalpha binding affinities (IC50) of 1.4 microM, 57 nM, and 53 nM. Importantly, they all exhibited ERalpha over ERbeta selectivity, with the most selective being 17-fold. The ligands also displayed low micomolar antiproliferative activity (7-15 microM) in the human MCF 7 breast cancer cell line. PMID- 17918821 TI - Interactions of hepatitis B core antigen and peptide inhibitors. AB - The core protein (HBcAg) of hepatitis B virus (HBV) has been shown to interact with the large surface antigen during HBV morphogenesis, and these interactions can be blocked by small peptides selected from either linear or constrained phage display peptide libraries. The association of HBcAg with peptide inhibitors was quantitatively evaluated by isothermal titration calorimetry. The thermodynamic data show that the interaction between HBcAg and peptide MHRSLLGRMKGA is enthalpy driven and occurs at a 3:1 stoichiometry and dissociation constant (Kd) value of 79.4 muM. However, peptide WSFFSNI displays a higher binding affinity for HBcAg with a Kd value of 18.5 muM when compared to peptide MHRSLLGRMKGA. A combinatorial approach using chemical cross-linking and surface-enhanced laser desorption/ionization-time-of-flight-mass spectrometry shows that the Lys of peptide MHRSLLGRMKGA interacted either with D64, E77, or D78 of HBcAg. PMID- 17918822 TI - Inhibitors of the interaction of a thyroid hormone receptor and coactivators: preliminary structure-activity relationships. AB - The modulation of gene regulation by blocking the interaction between the thyroid receptor (TR) and obligate coregulators has been reported recently with the discovery of the lead compound 3-(dimethylamino)-1-(4-hexylphenyl)propan-1-one). Herein we report studies aimed at optimization of this initial hit to determine the basic parameters of the structure-activity relationships and clarify the mechanism of action. These studies provided new insights, showing that activity and TRbeta isoform selectivity is highly correlated with the structural composition of these covalent inhibitors. PMID- 17918825 TI - Thermally induced two-step, two-site incomplete 6A1<-->2T2 crossover in a mononuclear iron(III) phenolate-pyridyl Schiff-base complex: a rare crystallographic observation of the coexistence of pure S=5/2 and 1/2 metal centers in the asymmetric unit. AB - The six-coordinate mononuclear iron(III) complexes [Fe(salpm)2]ClO(4).0.5EtOH, [Fe(salpm)2]Cl, [Fe{(3,5-tBu2)-salpm}2]X (X=ClO4- or Cl-), and [Fe{(3,5-tBu2) salpm}2]NO(3).2H2O [Hsalpm=N-(pyridin-2-ylmethyl)salicylideneamine; H(3,5-tBu2) salpm=N-(pyridin-2-ylmethyl)-3,5-di-tert-butylsalicylideneamine] have been synthesized and isolated in crystalline form; their chemical identities have been ascertained by elemental analyses, FAB mass spectrometry, and infrared spectroscopy. The room-temperature effective magnetic moments [(8chiMT)1/2 approximately 5.85-5.90 microB] of these complexes are consistent with the high spin (S=5/2) ground state. These complexes are intensely colored on account of the strong ppi-->dpi* LMCT visible absorptions. Definitive evidence for the structures of [Fe(salpm)2]ClO(4).0.5EtOH and [Fe{(3,5-tBu2)-salpm}2]NO(3).2H2O has been provided by single-crystal X-ray crystallography. The monomeric complex cations in both compounds comprise two uninegative phenolate-pyridyl tridentate Schiff-base ligands coordinated meridionally to the iron(III) to afford a distorted octahedral geometry with a trans,cis,cis-[FeO2N4] core. Whereas [Fe(salpm)2]ClO(4).0.5EtOH undergoes a thermally induced 6A1<-->2T2 crossover, [Fe{(3,5-tBu2)-salpm}2]NO(3).2H2O retains its spin state in the solid state down to 5 K. However, EPR spectroscopy reveals that the latter complex does exhibit a spin transformation in solution, albeit to a much lesser extent than does the former. The spin crossover in [Fe(salpm)2]ClO(4).0.5EtOH has resulted in an unprecedented crystallographic observation of the coexistence of high-spin and low-spin iron(III) complex cations in equal proportions around 100 K. At room temperature, the two crystallographically distinct ferric centers are both high spin; however, one [Fe(salpm)2]+ complex cation undergoes a complete spin transition over the temperature range approximately 200-100 K, whereas the other converts very nearly completely between 100 and 65 K; approximately 10% of the complex cations in [Fe(salpm)2]ClO(4).0.5EtOH remain in the high-spin state down to 5 K. PMID- 17918824 TI - Discovery of 1-[2-[(1S)-(3-dimethylaminopropionyl)amino-2-methylpropyl]-4 methylphenyl]-4-[(2R)-methyl-3-(4-chlorophenyl)-propionyl]piperazine as an orally active antagonist of the melanocortin-4 receptor for the potential treatment of cachexia. AB - A potent and selective antagonist of the melanocortin-4 receptor, 1-[2-[(1S)-(3 dimethylaminopropionyl)amino-2-methylpropyl]-6-methylphenyl]-4-[(2R)-methyl-3-(4 chlorophenyl)propionyl]piperazine (10d), was identified from a series piperazinebenzylamine attached with a N,N-dimethyl-beta-alanine side chain. This compound possessed high water solubility and exhibited good metabolic profiles. In animals, 10d showed moderate to good oral bioavailability and promoted food intake in tumor-bearing mice after oral administration. PMID- 17918823 TI - Small molecules block the polymerization of Z alpha1-antitrypsin and increase the clearance of intracellular aggregates. AB - The Z mutant of alpha1-antitrypsin (Glu342Lys) causes a domain swap and the formation of intrahepatic polymers that aggregate as inclusions and predispose the homozygote to cirrhosis. We have identified an allosteric cavity that is distinct from the interface involved in polymerization for rational structure based drug design to block polymer formation. Virtual ligand screening was performed on 1.2 million small molecules and 6 compounds were identified that reduced polymer formation in vitro. Modeling the effects of ligand binding on the cavity and re-screening the library identified an additional 10 compounds that completely blocked polymerization. The best antagonists were effective at ratios of compound to Z alpha1-antitrypsin of 2.5:1 and reduced the intracellular accumulation of Z alpha1-antitrypsin by 70% in a cell model of disease. Identifying small molecules provides a novel therapy for the treatment of liver disease associated with the Z allele of alpha1-antitrypsin. PMID- 17918827 TI - Self-assembly of a Mn9 nanoscopic mixed-valent cluster: synthesis, crystal structure, and magnetic behavior. AB - A rare Mn9 micro3-oxo-centered mixed-valent cluster [Mn9O7(O2CPh)11(thmn)(py)2 (H2O)3] (1) is prepared by assembling an oxo-centered MnIIMnIII2 triangle, [Mn3O(O2CPh)6(py)2(H2O)].0.5MeCN, as the secondary building unit in the presence of a tripodal alcohol, 1,1,1-tris(hydroxymethyl)nitromethane (H3thmn), as the capping ligand. Complex 1 was formed along with a minor byproduct, [Mn6O2(O2CPh)10(MeCN)4] (2). Complex 1 was characterized by X-ray single-crystal structure analysis and was crystallized in a monoclinic system, space group P2(1)/n, a=16.214(6) A, b=25.874(10) A, c=26.497(10) A, and beta=94.214(7) degrees. The Manganese-oxo-carboxylate core in 1 looks like a funnel. Variable temperature magnetic studies down to 2 K reveal the existence of dominant ferromagnetic interaction within the cluster. Alternating current susceptibility data of the cluster show strong frequency dependence of both the real and imaginary parts of susceptibility chi' and chi'' below 5 K. Moreover, the calculated relaxation time, tau0=1.2x10(-7) s, and the energy barrier, DeltaE=25 K, are consistent with the single-molecule magnetic behavior of 1. PMID- 17918826 TI - Iron(III) complexes with a tripodal N3O ligand containing an internal base as a model for catechol intradiol-cleaving dioxygenases. AB - A bis(mu-alkoxo)-bridged dinuclear iron(III) complex [Fe(L)(NO3)]2(NO3)2 [1; HL = N,N-bis(2-pyridylmethyl)-N-(2-hydroxyethyl)amine] of the tripodal N3O ligand was prepared as a biomimetic model for the intradiol-cleaving dioxygenase enzymes. The reaction of 1 and catechol in the presence of excess triethylamine gave the catecholate (CAT) chelate bis(mu-alkoxo)-bridged dinuclear iron(III) complex [Fe(L)(CAT)]2 (2). The molecular structures of complexes 1 and 2 were determined by X-ray crystallography. Diiron complexes 1 and 2 contain the same bis(mu alkoxo)diiron diamond core. All heteroatoms (N3O) of the ligand are coordinated to the iron center in complex 1 with two pyridine nitrogen atoms on the axial bonds, while one of the pyridyl arms of the ligand is left uncoordinated in complex 2. The interaction of the diiron complex 1 and 3,5-di-tert-butylcatechol (H2DBC) was investigated by electronic and mass spectroscopy. Complex 1 displays the intradiol-cleaving dioxygenase activity, and the coordinate ethoxyl arm of the ligand is capable of accepting the proton from catechol, which mimics the function of Tyr447 in the protocatechuate 3,4-dioxygenase as an internal base. The spectrophotometric titration experiment indicates the relatively low demand of the external base (0.8 equiv based on Fe(3+)) for attaining the highest dioxygenase activity of complex 1. The reaction rate of the reactive intermediate [Fe(HL)(DBC)]+ with dioxygen is 0.38 M(-1) s(-1) determined by kinetic studies. PMID- 17918829 TI - Synthesis and structure of new compounds with Zn-Ga bonds: insertion of the gallium(I) bisimidinate Ga(DDP) into Zn-X (X = CH3, Cl) and the homoleptic complex cation [Zn(GaCp*)4]2+. AB - Insertion reactions of the low-valent group 13 bisimidinate ligand Ga(DDP) {DDP = 2-[(2,6-diisopropylphenyl)amino]-4-[(2,6-diisopropylphenyl)imino]-2-pentene} into Zn-Me and Zn-Cl bonds are reported. The reaction of ZnMe2 with 2 equiv of Ga(DDP) yields the double-insertion product [{(DDP)GaMe}2Zn] (1), whereas the insertion of Ga(DDP) into the Zn-Cl bond of ZnCl2 in tetrahydrofuran (THF) leads to the monoinsertion product [{(DDP)GaCl}ZnCl(THF)2] (2). Treatment of 2 with Na[BArF] results in the salt [{THF.Ga(DDP)}Zn(THF)(mu-Cl)]2[BArF]2 (3), with two Cl atoms bridging the Zn centers. The structural features of the Zn-Ga-bonded compounds 1 3 were compared with related complexes and in particular with the compound [Zn(GaCp*)4][BArF]2 (4), which was synthesized by the reaction of ZnMe2, [H(OEt2)2][BArF], and GaCp* in fluorobenzene. The complex cation [Zn(GaCp*)4]2+ of 4 relates to previously reported d10 analogues [M(GaCp*)4] (M = Ni, Pd, Pt). All new compounds were fully characterized by elemental analysis, NMR spectroscopy, and single-crystal X-ray diffraction analysis. PMID- 17918828 TI - Synthesis, structural, and magnetic studies on a redox family of tetrametallic vanadium clusters: {VIV4}, {VIII2VIV2}, and {VIII4} butterfly complexes. AB - The synthesis, crystal structures, and magnetic properties are reported for a redox family of butterfly-type tetrametallic vanadium alkoxide clusters, namely [V2(VO)2(acac)4(RC{CH2O}3)2] (R=Me 1, Et 2, CH2OH 3), [V2(VO)2(acac)2(O2CPh)2(MeC{CH2O}3)2] (5), [(VO)4(MeOH)2(O2CPh)2({HOCH2}C{CH2O}3)2] (6), [V4Cl2(dbm)4(RC{CH2OH}3)2] (R=Me 7, Et 8, CH2OH 9), and [V4Cl2(dbm)4(MeO)6] (10). The cluster cores are {VIV4} (6), {VIII2VIV2} (1-5), and {VIII4} (7-10), with examples of both isomeric forms of the of the mixed-valence cores (either VIII or VIV ions forming the butterfly body). Magnetic studies reveal the clusters to be dominated by antiferromagnetic exchange interactions in each case. The magnetic exchange parameters are determined for representative examples of each core type. {VIV4} and {VIII4} have diamagnetic ground states. The two isomeric {VIII2VIV2} types are found to give rise to either an S=0 ground state with a number of low-lying excited states due to competing antiferromagnetic exchange interactions (VIII2 butterfly body) or to a well-isolated S=1 ground state (VIV2 butterfly body). PMID- 17918830 TI - Strategy to tether organometallic ruthenium-arene anticancer compounds to recombinant human serum albumin. AB - In order to utilize macromolecules for drug targeting and delivery, a strategy to tether organometallic ruthenium-arene drugs to carrier protein molecules was developed. The approach involves the design of a drug fragment capable of conjugating to linker molecules on a modified carrier protein via hydrazone bond formation. The proof-of-concept using recombinant human serum albumin is described. PMID- 17918831 TI - The sixteen CB(11)H(n)Me(12-n)(-) anions with fivefold substitution symmetry: anodic oxidation and electronic structure. AB - The 15 symmetrically methylated derivatives of the CB11H12(-) anion (1a) have been synthesized and found to vary greatly in ease of oxidation. Cyclic voltammetry in liquid SO2 yielded fully reversible oxidation potentials for five of those that have no adjacent unsubstituted vertices in positions 7-12; three others showed some indication of reversibility. The anions 1a-16a and the Jahn Teller distorted neutral radicals 1r-16r have been characterized by ab initio and density functional theory calculations. In the state average CASSCF(13,12)/6 31+G* approximation, the ground state potential energy surface of 1r contains five symmetry-related pairs of minima. The computational results account for the reversible redox potentials very well when the solvent is included explicitly (RI DFT(BP)/TZVP, COSMO). For display and for a semiquantitative understanding of methyl substituent effects in terms of perturbation theory, the molecular orbitals of 1a have been expressed in the symmetry-adapted cluster basis. The results serve as an underpinning for a set of additive empirical increments for redox potential prediction. Relative to the usual hydrogen standard, a single methyl group facilitates oxidation by approximately 50, 70, 70, and 10 mV in positions 1, 2, 7, and 12, respectively. This electron donor effect on the redox potential is due to a contribution, whereas those of (inductive and direct field) type are negligible. PMID- 17918832 TI - Polarized X-ray absorption spectroscopy of single-crystal Mn(V) complexes relevant to the oxygen-evolving complex of photosystem II. AB - High-valent Mn-oxo species have been suggested to have a catalytically important role in the water splitting reaction which occurs in the Photosystem II membrane protein. In this study, five- and six-coordinate mononuclear Mn(V) compounds were investigated by polarized X-ray absorption spectroscopy in order to understand the electronic structure and spectroscopic characteristics of high-valent Mn species. Single crystals of the Mn(V)-nitrido and Mn(V)-oxo compounds were aligned along selected molecular vectors with respect to the X-ray polarization vector using X-ray diffraction. The local electronic structure of the metal site was then studied by measuring the polarization dependence of X-ray absorption near-edge spectroscopy (XANES) pre-edge spectra (1s to 3d transition) and comparing with the results of density functional theory (DFT) calculations. The Mn(V)-nitrido compound, in which the manganese is coordinated in a tetragonally distorted octahedral environment, showed a single dominant pre-edge peak along the MnN axis that can be assigned to a strong 3d(z(2))-4p(z) mixing mechanism. In the square pyramidal Mn(V)-oxo system, on the other hand, an additional peak was observed at 1 eV below the main pre-edge peak. This component was interpreted as a 1s to 3d(xz,yz) transition with 4px,y mixing, due to the displacement of the Mn atom out of the equatorial plane. The XANES results have been correlated to DFT calculations, and the spectra have been simulated using a TD (time-dependent)-DFT approach. The relevance of these results to understanding the mechanism of the photosynthetic water oxidation is discussed. PMID- 17918833 TI - Pd-catalyzed amidation of aryl chlorides using monodentate biaryl phosphine ligands: a kinetic, computational, and synthetic investigation. AB - We present results on the amidation of aryl halides and sulfonates utilizing a monodentate biaryl phosphine-Pd catalyst. Our results are in accord with a previous report that suggests that the formation of kappa(2)-amidate complexes is deleterious to the effectiveness of a catalyst for this transformation and that their formation can be prevented by the use of appropriate bidentate ligands. We now provide data that suggest that the use of certain monodentate ligands can also prevent the formation of the kappa(2)-amidate complexes and thereby generate more stable catalysts for the amination of aryl chlorides. Furthermore, computational studies shed light on the importance of the key feature(s) of the biaryl phosphines (a methyl group ortho to the phosphorus center) that enable the coupling to occur. The use of ligands that possess a methyl group ortho to the phosphorus center allows a variety of aryl and heteroaryl chlorides with various amides to be coupled in high yield. PMID- 17918835 TI - Aromatic molecules in anion recognition: electrostatics versus H-bonding. AB - Mass-selected complexes A-.C6FnH(6-n) (A = Cl, I, SF6; n = 0-5) were studied by infrared photodissociation spectroscopy and computational chemistry methods to investigate the interaction of negative ions and aromatic molecules, in which the charge distribution can be tuned by fluorination. Surprisingly, we find that, despite positive partial charges on the carbon atoms at high levels of fluorination, all anions under study prefer hydrogen bonding to the remaining H atoms in the ligand rather than binding to the positively charged ring. Moreover, bifurcated hydrogen bonds to two neighboring CH groups are energetically favored over linear hydrogen bonds to a single CH group. PMID- 17918834 TI - Synthetic efficiency in enzyme mechanisms involving carbocations: aristolochene synthase. AB - An intramolecular proton-transfer mechanism has been proposed for the carbocationic cyclization of farnesyl pyrophosphate (FPP) to (+)-aristolochene catalyzed by aristolochene synthase. This novel mechanism, which is based on results obtained by high-level ab initio molecular orbital and density functional theory calculations, differs from the previous proposal in the key step of carbocation propagation prior to the formation of the bicyclic carbon skeleton. Previously, germacrene A was proposed to be generated as an intermediate by deprotonation of germacryl cation followed by reprotonation of the C6-C7 double bond to yield eudesmane cation. In the mechanism proposed here the direct intramolecular proton transfer has a computed barrier of about 22 kcal/mol, which is further lowered to 16-20 kcal/mol by aristolochene synthase. An alternative pathway is also possible through a proton shuttle via a pyrophosphate-bound water molecule. The mechanism proposed here is consistent with the observation that germacrene A is not a substrate of aristolochene synthase. Furthermore, the modeled substrate-enzyme complex suggests that Trp 334 and Phe 178 play key roles in positioning the substrate in the reactive orientation in the binding pocket. This is consistent with experimental findings that mutations of either residue lead to pronounced generation of aborted cyclization products. PMID- 17918836 TI - Synchronicity of mononuclear and dinuclear events in homogeneous catalysis. Hydroformylation of cyclopentene using Rh4(CO)12 and HRe(CO)5 as precursors. AB - The combined application of two or more metals in homogeneous catalysis can lead to synergistic effects; however, the phenomenological basis for these observations often goes undetermined. The hetero-bimetallic catalytic binuclear elimination reaction, a system involving both mononuclear and dinuclear intermediates, has been repeatedly suggested as a possible mechanism. In the present contribution, the simultaneous application of Rh4(CO)12 and HRe(CO)5 as precursors in the hydroformylation reaction leads to a very strong synergistic rate effect. In situ spectroscopic measurements confirm the presence of both mononuclear and dinuclear intermediates such as RCORh(CO)4 and RhRe(CO)9 in the active system. Moreover, kinetic analysis confirms interconversion of these intermediates as well as their statistical correlation with organic product formation. Specifically, the rate of hydrogen activation by RhRe(CO)9 is exactly equal to the rate of aldehyde formation from binuclear elimination between HRe(CO)5 and RCORh(CO)4 at all reaction conditions studied. Thus the catalytic events involving mononuclear species and those involving dinuclear species are synchronized. In the present experiments, the new topology is orders of magnitude more efficient than the corresponding unicyclic rhodium system. PMID- 17918837 TI - Conformational manifold of alpha-aminoisobutyric acid (Aib) containing alanine based tripeptides in aqueous solution explored by vibrational spectroscopy, electronic circular dichroism spectroscopy, and molecular dynamics simulations. AB - Replacement of the alpha-proton of an alanine residue to generate alpha aminoisobutyric acid (Aib) in alanine-based oligopeptides favors the formation of a 3(10) helix when the length of the oligopeptide is about four to six residues. This research was aimed at experimentally identifying the structural impact of an individual Aib residue in an alanine context of short peptides in water and Aib's influence on the conformation of nearest-neighbor residues. The amide I band profile of the IR, isotropic and anisotropic Raman, and vibrational circular dichroism (VCD) spectra of Ac-Ala-Ala-Aib-OMe, Ac-Ala-Aib-Ala-OMe, and Ac-Aib-Ala Ala-OMe were measured and analyzed in terms of different structural models by utilizing an algorithm that exploits the excitonic coupling between amide I' modes. The conformational search was guided by the respective 1H NMR and electronic circular dichroism spectra of the respective peptides, which were also recorded. From these analyses, all peptides adopted multiple conformations. Aib predominantly sampled the right-handed and left-handed 3(10)-helix region and to a minor extent the bridge region between the polyproline (PPII) and the helical regions of the Ramachandran plot. Generally, alanine showed the anticipated PPII propensity, but its conformational equilibrium was shifted towards helical conformations in Ac-Aib-Ala-Ala-OMe, indicating that Aib can induce helical conformations of neighboring residues positioned towards the C-terminal direction of the peptide. An energy landscape exploration by molecular dynamics simulations corroborated the results of the spectroscopic studies. They also revealed the dynamics and pathways of potential conformational transitions of the corresponding Aib residues. PMID- 17918838 TI - Spectroscopic and kinetic studies of perturbed trinuclear copper clusters: the role of protons in reductive cleavage of the O-O bond in the multicopper oxidase Fet3p. AB - The multicopper oxidase Fet3p couples four 1e(-) oxidations of substrate to the 4e(-) reduction of O2 to H2O. Fet3p uses four Cu atoms to accomplish this reaction: the type 1, type 2, and coupled binuclear type 3 sites. The type 2 and type 3 sites together form a trinuclear Cu cluster (TNC) which is the site of O2 reduction. This study focuses on mutants of two residues, E487 and D94, which lie in the second coordination sphere of the TNC and defines the role that each plays in the structural integrity of the TNC, its reactivity with O2, and in the directional movement of protons during reductive cleavage of the O-O bond. The E487D, E487A, and D94E mutants have been studied in the holo and type 1 depleted (T1D) forms. Residue E487, located near the T3 center, is found to be responsible for donation of a proton during the reductive cleavage of the O-O bond in the peroxide intermediate and an inverse kinetic solvent isotope effect, which indicates that this proton is already transferred when the O-O bond is cleaved. Residue D94, near the T2 site, plays a key role in the reaction of the reduced TNC with O2 and drives electron transfer from the T2 Cu to cleave the O-O bond by deprotonating the T2 Cu water ligand. A mechanism is developed where these second sphere residues participate in the proton assisted reductive cleavage of the O-O bond at the TNC. PMID- 17918840 TI - Meta conjugation effect on the torsional motion of aminostilbenes in the photoinduced intramolecular charge-transfer state. AB - The photochemical behavior of a series of trans-3-(N-arylamino)stilbenes (m1, aryl = 4-substituted phenyl with a substituent of cyano (CN), hydrogen (H), methyl (Me), or methoxy (OM)) in both nonpolar and polar solvents is reported and compared to that of the corresponding para isomers (p1CN, p1H, p1Me, and p1OM). The distinct propensity of torsional motion toward a low-lying twisted intramolecular charge-transfer (TICT) state from the planar ICT (PICT) precursor between the meta and para isomers of 1CN and 1Me reveals the intriguing meta conjugation effect and the importance of the reaction kinetics. Whereas the poor charge-redistribution (delocalization) ability through the meta-phenylene bridge accounts for the unfavorable TICT-forming process for m1CN, it is such a property that slows down the decay processes of fluorescence and photoisomerization for m1Me, facilitating the competition of the single-bond torsional reaction. In contrast, the quinoidal character for p1Me in the PICT state kinetically favors both fluorescence and photoisomerization but disfavors the single-bond torsion. The resulting concept of thermodynamically allowed but kinetically inhibited TICT formation could also apply to understanding the other D-A systems, including trans-4-cyano-4'-(N,N-dimethylamino)stilbene (DCS) and 3-(N,N dimethylamino)benzonitrile (3DMABN). PMID- 17918841 TI - Carbon dioxide-mediated catalytic rearrangement of propargyl alcohols into alpha,beta-unsaturated ketones. PMID- 17918842 TI - TROSY-based z-exchange spectroscopy: application to the determination of the activation energy for intermolecular protein translocation between specific sites on different DNA molecules. AB - A two-dimensional TROSY-based z-exchange 1H-15N correlation experiment for the quantitative analysis of kinetic processes in the slow exchange regime is presented. The pulse scheme converts the product operator terms Nz into 2NzHz and 2NzHz into -Nz in the middle of the z-mixing period, thereby suppressing the buildup of spurious semi-TROSY peaks arising from the different relaxation rates for the Nz and 2NzHz terms and simplifying the behavior of longitudinal magnetization for an exchanging system during the mixing period. Theoretical considerations and experimental data demonstrate that the TROSY-based z-exchange experiment permits quantitative determination of rate constants using the same procedure as that for the conventional non-TROSY 15Nz-exchange experiment. Line narrowing as a consequence of the use of the TROSY principle makes the method particularly suitable for kinetic studies at low temperature, thereby permitting activation energies to be extracted from data acquired over a wider temperature range. We applied this method to the investigation of the process whereby the HoxD9 homeodomain translocates between specific target sites on different DNA molecules via a direct transfer mechanism without going through the intermediary of free protein. The activation enthalpy for intermolecular translocation was determined to be 17 kcal/mol. PMID- 17918839 TI - Electronic structure of the peroxy intermediate and its correlation to the native intermediate in the multicopper oxidases: insights into the reductive cleavage of the o-o bond. AB - The multicopper oxidases (MCOs) utilize a blue type 1 (T1) copper site and a trinuclear Cu cluster composed of a type 2 (T2) and a binuclear type 3 (T3) site that together catalyze the four-electron reduction of O2 to H2O. Reaction of the fully reduced enzyme with O2 proceeds via two sequential two-electron steps generating the peroxy intermediate (PI) and the native intermediate (NI). While a detailed description of the geometric and electronic structure of NI has been developed, this has been more elusive for PI largely due to the diamagnetic nature of its ground state. Density functional theory (DFT) calculations have been used to correlate to spectroscopic data to generate a description of the geometric and electronic structure of PI. A highly conserved carboxylate residue near the T2 site is found to play a critical role in stabilizing the PI structure, which induces oxidation of the T2 and one T3 Cu center and strong superexchange stabilization via the peroxide bridge, allowing irreversible binding of O2 at the trinuclear Cu site. Correlation of PI to NI is achieved using a two-dimensional potential energy surface generated to describe the catalytic two-electron reduction of the peroxide O-O bond by the MCOs. It is found that the reaction is thermodynamically driven by the relative stability of NI and the involvement of the simultaneous two-electron-transfer process. A low activation barrier (calculated approximately 5-6 kcal/mol and experimental approximately 3-5 kcal/mol) is produced by the triangular topology of the trinuclear Cu cluster site, as this symmetry provides good donor-acceptor frontier molecular orbital (FMO) overlap. Finally, the O-O bond cleavage in the trinuclear Cu cluster can be achieved via either a proton-assisted or a proton unassisted process, allowing the MCOs to function over a wide range of pH. It is found that while the proton helps to stabilize the acceptor O22- sigma* orbital in the proton-assisted process for better donor-acceptor FMO overlap, the third oxidized Cu center in the trinuclear site assumes the role as a Lewis acid in the proton-unassisted process for similarly efficient O-O bond cleavage. PMID- 17918843 TI - 7,8-dihydro retinals outperform the native retinals in conferring photosensitivity to visual opsin. AB - The visual pigment rhodopsin presents an astonishing photochemical performance. It exhibits an unprecedented quantum yield (0.67) in a highly defined and ultrafast photoisomerization process. This triggers the conformational changes leading to the active state Meta II of this G protein-coupled receptor. The responsible ligand, retinal, is covalently bound to Lys-296 of the protein in a protonated Schiff base. The resulting positive charge delocalization over the terminal part of the polyene chain of retinal creates a conjugation defect that upon photoexcitation moves to the opposite end of the polyene. Shortening the polyene as in 5,6-dihydro- or 7,8-dihydro analogues might facilitate photoisomerization of a 9-Z and an 11-Z bond. Here we describe pigment analogues generated with bovine opsin and 11-Z 7,8-dihydro retinal or 9-Z 7,8-dihydro retinal. Both isomers readily generate photosensitive pigments that differ remarkably in spectral properties from the native pigments. In addition, in spite of the more flexible 7,8 single bond, both analogue pigments exhibit strikingly efficient photoisomerization while largely maintaining the activity toward the G protein. These results bear upon the activation of ligand-gated signal transducers such as G protein-coupled receptors. PMID- 17918844 TI - Diversification of self-organized architectures in supramolecular dye assemblies. AB - Upon complexation with bismelamine receptors (BMn) featuring different alkyl linker lengths (number of methylene groups (n) = 5-12), a barbituric acid merocyanine dye (1) can be loaded into diverse self-organized superstructures through multiple hydrogen-bonding interactions. UV/vis, dynamic light scattering, and NMR studies in cyclohexane demonstrate that the diversification of the primarily formed hydrogen-bonded species in solution occurs by varying the linker length of BMn. Hierarchical organization of the hydrogen-bonded species is achieved by slow evaporation of the solvent (forming solvent-free films), and the resulting superstructures are evaluated by polarized optical microscopy, X-ray diffraction, SEM, and AFM techniques. The formation of columnar structures with and without two-dimensional ordering are revealed for shorter (n = 5-7) and longer (n = 11, 12) linker bis(melamines), respectively. On the contrary, in the cases of n = 8-10, the formation of lamellar structures is unveiled. Several assemblies (n = 5, 7, 11) indicate the formation of a liquid crystalline mesophase in POM and DSC analyses. Hierarchical organization is also achieved in solution by prolonged aging, affording phase-separated crystalline nanofibers (n = 5, 7) and soft nanofibrils agglomerating into wormlike objects (n = 8), gel forming continuous globular networks (n = 10), and nanofibers (n = 11, 12). These superstructural and morphological diversifications are an outcome of the variation in the primarily formed hydrogen-bonded supramolecular architectures. Using this strategy, diverse self-assembled materials will be obtained from a single dye component. PMID- 17918845 TI - Exchange-biased dimers of single-molecule magnets in OFF and ON states. PMID- 17918846 TI - Reaction of [sigma:eta(5)-(C(9)H(6))C(2)B(9)H(10)]Zr(NMe(2))(DME) with guanidines: metallacarborane-mediated C-N bond cleavage and 1,5-sigmatropic rearrangement. PMID- 17918847 TI - Cylindrical block co-micelles with spatially selective functionalization by nanoparticles. PMID- 17918848 TI - Trisubstituted isoalloxazines as a new class of G-quadruplex binding ligands: small molecule regulation of c-kit oncogene expression. PMID- 17918849 TI - Product deuterium isotope effect for orotidine 5'-monophosphate decarboxylase: evidence for the existence of a short-lived carbanion intermediate. PMID- 17918850 TI - Copper(II)-catalyzed enantioselective intramolecular carboamination of alkenes. PMID- 17918855 TI - Grignard reagents: alkoxide-directed iodine-magnesium exchange at sp3 centers. AB - Sequential addition of i-PrMgCl and BuLi to sp3 hybridized iodoalcohols triggers a facile iodine-metal exchange. Intercepting the resulting cyclic Grignard reagents with a slight excess of an electrophile leads to a diverse range of substituted alcohols. The iodine-magnesium exchange strategy is effective with 3 carbon iodoalcohols bearing alkyl substitutents on the carbinol or adjacent carbons and with the chain-extended homolog 4-iodobutan-1-ol. PMID- 17918854 TI - Synthetic modification of manzamine A via Grubbs metathesis. Novel structures with enhanced antibacterial and antiprotozoal properties. AB - A strategy for the structural modification of biologically important alkene containing natural products via ring-opening olefin metathesis is described. Exposure of manzamine A 1 to the second-generation Grubbs catalyst in the presence of ethylene leads to the formation of 2 and 4. The antibacterial activity of the novel manzamine analogue 2 (IC50=0.10 nM) against Mycobacterium intracellulare is ca. 2-fold more potent than that of ciprofloxacin (IC50=0.18 nM), a drug that is frequently used against antibiotic-resistant infections. PMID- 17918856 TI - Nitric oxide prodrugs: diazeniumdiolate anions of hindered secondary amines. AB - Nitric oxide prodrugs derived from hindered secondary amines were prepared. The decomposition patterns of these prodrugs indicate that alpha-methyl groups around the nitrogen bearing the diazeniumdiolate group prolong their half-life in aqueous buffer. PMID- 17918858 TI - Analysis of mechanistic pathway models in drug discovery: p38 pathway. AB - Mechanistic models of signal transduction have emerged as valuable tools for untangling complex signaling networks and gaining detailed insight into pathway dynamics. The natural extension of these tools is for the design of therapeutic strategies. We have generated a novel computational model of lipopolysaccharide induced p38 signaling in the context of TNF-alpha production in inflammatory disease. Using experimental measurement of protein levels and phospho-protein time courses, populations of model parameters were estimated. With a collection of parameter sets, reflecting virtual diversity, we step through analysis of the p38 signaling pathway model to answer specific drug discovery questions regarding target prioritization, inhibitor simulation, model robustness and co-drugging. We demonstrate that target selection cannot be assessed independently from inhibitor mechanism of action and is also linked with robustness to cellular variability. Finally, we assert that in the face of parameter uncertainty one can still uncover consistent findings that can guide drug discovery efforts. PMID- 17918859 TI - Protein binding to amphoteric polymer brushes grafted onto a porous hollow-fiber membrane. AB - Three kinds of ampholites, i.e., 3-aminopropionic acid (NH2C2H4COOH), (2 aminoethyl)phosphonic acid (NH2C2H4PO3H2), and 2-aminoethane-1-sulfonic acid (NH2C2H4SO3H), were introduced into an epoxy group-containing polymer brush grafted onto a porous hollow-fiber membrane with a porosity of 70% and pore size of 0.36 microm. The amphoteric group density of the hollow-fiber ranged from 0.50 to 0.72 mmol/g. Three kinds of proteins, i.e., lactoferrin (Lf), cytochrome c (Cyt c), and lysozyme (Ly), were captured by the amphoteric polymer brush during the permeation of the protein solution across the ampholite-immobilized porous hollow-fiber membrane. Multilayer binding of the protein to the amphoteric polymer brush, with a degree of multilayer binding of 3.3, 8.6, and 15 for Lf, Cyt c, and Ly, respectively, with the (2-aminoethyl)phosphonic acid-immobilized porous hollow-fiber membrane, was demonstrated with a negligible diffusional mass transfer resistance of the protein to the ampholite immobilized. The 2 aminoethane-1-sulfonic acid-immobilized porous hollow-fiber membrane exhibited the lowest initial flux of the protein solution, 0.41 m/h at a transmembrane pressure of 0.1 MPa and 298 K, and the highest equilibrium binding capacity of the protein, e.g., 130 mg/g for lysozyme. Extension and shrinkage of the amphoteric polymer brushes were observed during the binding and elution of the proteins. PMID- 17918860 TI - Method for profiling mucin oligosaccharides from gastric biopsies of rhesus monkeys with and without Helicobacter pylori infection. AB - A method for in vivo analysis of gastric mucin oligosaccharides was developed and applied to rhesus monkeys with and without Helicobacter pylori infection. Mucin type O-linked oligosaccharides were directly released by reductive beta elimination from gastric biopsies from rhesus monkeys. The released oligosaccharides were structurally characterized by matrix-assisted laser desorption/ionozation and electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. A diverse profile of neutral and acidic oligosaccharides was observed with these techniques. The most predominant core structure detected in all of the samples at relatively high abundance corresponded to core 2 (2HexNAc-1Hex, m/z = 611.227). The spectra generated from H. pylori-infected monkey samples showed fewer oligosaccharides collectively. Peaks corresponding to 1HexNAc-1Hex (m/z = 408.148) and 2HexNAc (m/z = 449.174), which most likely represent core structures, were absent in all infected monkeys studied, although present in all uninfected monkeys. Unsupervised cluster analysis demonstrated clear differences between the peaks detected in uninfected and naturally infected monkey samples. The results suggest that H. pylori infection is associated with lower relative abundance of oligosaccharides and loss of mucin-type core structures. This method can be applied to characterize the glycans associated with the mucin lining of live animals and allows for repeated analysis of the same animal over the course of infection. PMID- 17918861 TI - Kinetic model for the ATP-dependent translocation of Saccharomyces cerevisiae RSC along double-stranded DNA. AB - The chromatin remodeling complex RSC from Saccharomyces cerevisiae is a DNA translocase that moves with directionality along double-stranded DNA in a reaction that is coupled to ATP hydrolysis. To better understand how this basic molecular motor functions, a novel method of analysis has been developed to study the kinetics of RSC translocation along double-stranded DNA. The data provided are consistent with RSC translocation occurring through a series of repeating uniform steps with an overall processivity of P = 0.949 +/- 0.003; this processivity corresponds to an average translocation distance of 20 +/- 1 base pairs (bp) before dissociation. Interestingly, a slow initiation process, following DNA binding, is required to make RSC competent for DNA translocation. These results are further discussed in the context of previously published studies of RSC and other DNA translocases. PMID- 17918862 TI - Effects of histidine protonation and phosphorylation on histidine-containing phosphocarrier protein structure, dynamics, and physicochemical properties. AB - Previous structural studies of the histidine-containing phosphocarrier protein (HPr) have shown that active site residue His15 can adopt two distinct conformations which were termed OPEN and CLOSED. Using molecular dynamics simulations and protonation probability calculations, we were able to show that these two conformations correspond to different protonation forms of the histidine ring. The CLOSED-to-OPEN transition requires His15 to adopt a conformation with higher energy, which is compensated by the favorable energetic consequences of protonation. Calculations of the conformational energy of His15 show that HPr exists mainly in the CLOSED form at pH 7. The very low apparent pKa value (3.2-4.5) of the CLOSED conformation and the fact that the imidazole ring of residue 15 is primarily unprotonated at Ndelta1 at neutral pH ensure that His15 is ideally primed to be specifically phosphorylated at Ndelta1. In contrast to unphosphorylated HPr, the phosphorylated form exhibits no conformational transitions, and the CLOSED state is stable even for the protonated imidazole ring due to favorable interactions between the phosphate group and the backbone of Ala16 and Arg17. These observations from MD simulations are confirmed by a simple four-microstate model which can explain both the pH-dependent conformational change of unphosphorylated HPr and the conformational rigidity of phosphorylated HPr. Our study suggests that the predominant CLOSED conformation is relevant for HPr function in the phosphotransfer reaction, while the OPEN form of unphosphorylated HPr might be important for its additional regulatory function, in which an OPEN conformation of His15 is recognized by the transcriptional regulator CcpA. PMID- 17918863 TI - Issues of ligand accessibility and mobility in initial cell attachment. AB - The influence of lateral ligand mobility on cell attachment and receptor clustering has previously been explored for membrane-anchored molecules involved in cell-cell adhesion. In this study, we considered instead a cell binding motif from the extracellular matrix. Even though the lateral mobility of extracellular matrix ligands in membranes does not occur in vivo, we believe it is of interest for cell engineering in vitro. As is the case for cell-cell adhesion molecules, lateral mobility of extracellular matrix ligands could influence cell attachment and, subsequently, cell behavior in cell culture. In this paper, the accessibility and functionality of extracellular matrix ligands presented at surfaces were evaluated for the conditions of laterally mobile versus non-mobile ligands by studying ligand-antibody binding events and early cell attachment as a function of ligand concentration. We compare the initial attachment of rat derived adult hippocampal progenitor (AHP) cells on laterally mobile, supported phospholipid bilayer membranes to non-mobile, poly-L-lysine-grafted-poly(ethylene glycol) (PLL-g-PEG) polymer films functionalized with a range of laminin-derived IKVAV-containing peptide densities. To this end, synthesis of a new PLL-g-PEG/PEG IKVAV polymer is described. The characterization of available IKVAV peptides on both surface presentations schemes was explored by studying the mass uptake of anti-IKVAV antibodies using a combination of the surface-sensitive techniques quartz crystal microbalance with dissipation monitoring, surface plasmon resonance spectroscopy, and optical waveguide lightmode spectroscopy. IKVAV containing peptides presented on laterally mobile, supported phospholipid bilayers and non-mobile PLL-g-PEG were recognized by the anti-IKVAV antibody in a dose-dependent manner, indicating that the amount of available IKVAV ligands increases proportionally with ligand density over the concentrations tested. Attachment of AHP cells to IKVAV-functionalized PLL-g-PEG and supported phospholipid bilayers followed a sigmoidal dependence on peptide concentration, with a critical concentration of approximately 3 pmol/cm2 IKVAV ligands required to support initial AHP cell attachment for both surface modifications. There appeared to be little influence of IKVAV peptide mobility on the initial attachment of AHP cells. Although the spread in the cell attachment data was larger for the PLL-g-PEG surface modification, this was reduced when observed after 24 h, indicating that the cells might need longer times to establish attachment strengths equivalent to those observed on peptide-functionalized supported lipid bilayers. The present study is a step toward understanding the influence of extracellular-matrix-derived ligand mobility on cell fate. Further analysis should focus on the systematic tuning of lateral ligand diffusion, as well as a comparison between the response of non-spreading cells (i.e., AHPs), versus spreading cells (i.e., fibroblasts). PMID- 17918864 TI - Flow-focusing generation of monodisperse water droplets wrapped by ionic liquid on microfluidic chips: from plug to sphere. AB - Generating droplets via microfluidic chips is a promising technology in microanalysis and microsynthesis. To realize room-temperature ionic liquid (IL) water two-phase studies in microscale, a water-immiscible IL was employed as the continuous phase for the first time to wrap water droplets (either plugs or spheres) on flow-focusing microfluidic chips. The IL, 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF6]), could wet both hydrophilic and hydrophobic channel surfaces because of its dual role of hydrophilicity/hydrophobicity and extremely high viscosity, thus offering the possibility of wrapping water droplets in totally hydrophilic (THI), moderately hydrophilic (MHI), and hydrophobic (HO) channels. The droplet shape could be tuned from plug to sphere, with the volume from 6.3 nL to 65 pL, by adding an orifice in the focusing region, rendering the hydrophilic channel surface hydrophobic, and suppressing the Uw/UIL ratio below 1.0. Three different breakup processes were defined and clarified, in which the sub-steady breakup and steady breakup were essential for the formation of plugs and spheric droplets, respectively. The influences of channel hydrophilicity/hydrophobicity on droplet formation were carefully studied by evaluating the wetting abilities of water and IL on different surfaces. The superiority of IL over water in wetting hydrophobic surface led to the tendency of forming small, spheric aqueous droplets in the hydrophobic channel. This IL favored droplet-based system represented a high efficiency in water/IL extraction, in which rhodamine 6G was extracted from aqueous droplets to [BMIM][PF6] in the hydrophobic orifice-included (HO-OI) channel in 0.51 s. PMID- 17918865 TI - Repulsion between oppositely charged macromolecules or particles. AB - The interaction of two oppositely charged surfaces has been investigated using Monte Carlo simulations and approximate analytical methods. When immersed in an aqueous electrolyte containing only monovalent ions, two such surfaces will generally show an attraction at large and intermediate separations. However, if the electrolyte solution contains divalent or multivalent ions, then a repulsion can appear at intermediate separations. The repulsion increases with increasing concentration of the multivalent salt as well as with the valency of the multivalent ion. The addition of a second salt with only monovalent ions magnifies the effect. The repulsion between oppositely charged surfaces is an effect of ion-ion correlations, and it increases with increasing electrostatic coupling and, for example, a lowering of the dielectric permittivity enhances the effect. An apparent charge reversal of the surface neutralized by the multivalent ion is always observed together with a repulsion at large separation, whereas at intermediate separations a repulsion can appear without charge reversal. The effect is hardly observable for a symmetric multivalent salt (e.g., 2:2 or 3:3). PMID- 17918866 TI - Monte Carlo simulation of mixed lennard-jones nonionic surfactant adsorption at the liquid/vapor interface. AB - New Monte Carlo simulations are presented for nonionic surfactant adsorption at the liquid/vapor interface of a monatomic solvent specifically investigating the roles of tail attraction and binary mixtures of different tail lengths. Surfactant molecules consist of an amphiphilic chain with a solvophilic head and a solvophobic tail. All molecules in the system, solvent and surfactant, are characterized by the Lennard-Jones (LJ) potential. Adjacent atoms along the surfactant chain are connected by finitely extensible harmonic springs. Solvent molecules move via the Metropolis random-walk algorithm, whereas surfactant molecules move according to the continuum configurational bias Monte Carlo (CBMC) method. We generate thermodynamic adsorption and surface-tension isotherms and compare results quantitatively to single-surfactant adsorption (Langmuir, 2007, 23, 1835). Surfactant tail groups with attractive interaction lead to cooperative adsorption at high surface coverage and higher maximum adsorption at the interface than those without. Moreover, adsorption and surface-tension isotherms with and without tail attraction are identical at low concentrations, deviating only near maximum coverage. Simulated binary mixtures of surfactants with differing lengths give intermediate behavior between that of the corresponding single-surfactant adsorption and surface-tension isotherms both with and without tail attraction. We successfully predict simulated mixture results with the thermodynamically consistent ideal adsorbed solution (IAS) theory for binary mixtures of unequal-sized surfactants using only the simulations from the single surfactants. Ultimately, we establish that a coarse-grained LJ surfactant system is useful for understanding actual surfactant systems when tail attraction is important and for unequal-sized mixtures of amphiphiles. PMID- 17918868 TI - An approximate model for the adhesive contact of rough viscoelastic surfaces. AB - Surface roughness is known to easily suppress the adhesion of elastic surfaces. Here, a simple model for the contact of viscoelastic rough surfaces with significant levels of adhesion is presented. This approach is derived from our previous model (Barthel, E.; Haiat, G. Langmuir 2002, 18, 9362) for the adhesive contact of viscoelastic spheres. For simplicity, a simple loading/unloading history (infinitely fast loading and constant pull-out velocity) is assumed. The model provides approximate analytical expressions for the asperity response and exhibits the full viscoelastic adhesive contact phenomenology such as stress relaxation inside the contact zone and creep at the contact edges. Combining this model with a Greenwood-Williamson statistical modeling of rough surfaces, we propose a quantitative assessment of the adhesion to rough viscoelastic surfaces. We show that moderate viscoelasticity efficiently restores adhesion on rough surfaces over a wide dynamic range. PMID- 17918869 TI - Open-Bench Method for the Preparation of BaTiO3, SrTiO3, and BaxSr1-xTiO3 nanocrystals at 80 degrees C. AB - In this letter, we report the first facile open-bench synthesis of BaTiO3, SrTiO3 nanocrystals, and their nanosolid solutions BaxSr1-xTiO3 (BST) at 80 degrees C. The size of the BST nanoparticles was readily tuned from approximately 50 to approximately 10 nm with achievable giant dielectric constants. The process yielded these important perovskite mixed-metal oxide crystals of high quality on the nanometer scale without a history of thermal stress. This new synthesis system involves inorganic starting materials without organic components and does not require demanding conditions such as an inert environment, high pressure, and high temperature. PMID- 17918867 TI - Micellization of dissymmetric cationic gemini surfactants and their interaction with dimyristoylphosphatidylcholine vesicles. AB - The micellization process of a series of dissymmetric cationic gemini surfactants [CmH2m+1(CH3)2N(CH2)6N(CH3)2C6H13]Br2 (designated as m-6-6 with m = 12, 14, and 16) and their interaction with dimyristoylphosphatidylcholine (DMPC) vesicles have been investigated. In the micellization process of these gemini surfactants themselves, critical micelle concentration (cmc), micelle ionization degree, and enthalpies of micellization (DeltaHmic) were determined, from which Gibbs free energies of micellization (DeltaGmic) and entropy of micellization (DeltaSmic) were derived. These properties were found to be influenced significantly by the dissymmetry in the surfactant structures. The phase diagrams for the solubilization of DMPC vesicles by the gemini surfactants were constructed from calorimetric results combining with the results of turbidity and dynamic light scattering. The effective surfactant to lipid ratios in the mixed aggregates at saturation (Resat) and solubilization (Resol) were derived. For the solubilization of DMPC vesicles, symmetric 12-6-12 is more effective than corresponding single-chain surfactant DTAB, whereas the dissymmetric m-6-6 series are more effective than symmetric 12-6-12, and 16-6-6 is the most effective. The chain length mismatch between DMPC and the gemini surfactants may be responsible for the different Re values. The transfer enthalpy per mole of surfactant within the coexistence range may be associated with the total hydrophobicity of the alkyl chains of gemini surfactants. The transfer enthalpies of surfactant from micelles to bilayers are always endothermic due to the dehydration of headgroups and the disordering of lipid acyl chain packing during the vesicle solubilization. PMID- 17918870 TI - Molecular dynamics simulations of capillary rise experiments in nanotubes coated with polymer brushes. AB - The capillary filling of a nanotube coated with a polymer brush is studied by molecular dynamics simulations of a coarse-grained model, assuming various conditions for the fluid-wall and fluid-brush interactions. Whereas the fluid is modeled by simple point particles interacting with Lennard-Jones forces, the (end grafted, fully flexible) polymers that form the brush coating are described by a standard bead-spring model. Our experiments reveal that capillary filling is observed even for walls that would not be wetted by the fluid, provided the polymer brush coating itself wets. Generally, it is found that the capillary rise always proceeds through a t1/2 law with time t while the underlying molecular mechanism differs for wettable and nonwettable walls. For wettable walls, fluid imbibition is compatible with the Lucas-Washburn mechanism whereby the total influx of matter drops steadily with growing chain length N and the meniscus speed goes through a minimum at intermediate chain lengths. Moreover, because of flow, the polymer brush reorganizes its structure by forming a dense plug of chain segments under the meniscus that follows the meniscus in its motion. When the tube wall does not wet, one observes no meniscus formation for short chains although the fluid seeps through the wet brush. For a brush coating with longer chains, axial segregation between the brush segments and the fluid occurs by a kind of diffusive spreading, reminiscent of invasion percolation transport in a random medium, leading to the formation of a moving meniscus. For even longer chains that reach the tube axis, the rise of a meniscus with vanishing curvature like imbibition in a porous medium is observed to take place. PMID- 17918871 TI - Temperature- and light-responsive blends of pluronic F127 and poly(N,N dimethylacrylamide-co-methacryloyloxyazobenzene). AB - Photoresponsive poly(N,N-dimethylacrylamide-co-methacryloyloxyazobenzene) (DMA MOAB) and temperature-responsive Pluronic F127 (F127) copolymers were blended to obtain systems responsive to both stimuli that are potentially useful for pharmaceutical formulations. The random DMA-MOAB copolymer undergoes a trans to cis isomerization when irradiated by 366 nm light, which modifies both the air water interfacial behavior and the self-associative properties of the copolymer. Under dark conditions the azobenzene groups of DMA-MOAB in the trans conformation self-associate and the interactions with F127 are minimal. The cis conformation of the azobenzene groups of the DMA-MOAB copolymer is relatively more hydrophilic than the trans conformation, which causes the copolymer micelles to dissociate upon irradiation, allowing the unimers to form mixed micelles with the F127. This causes the sol-gel transition temperature of the DMA-MOAB:F127 blend to be 10 degrees C lower upon irradiation at 366 nm compared to that for the dark conditions. It has been found that F127 (10-12 wt %):DMA-MOAB (5-6 wt %) aqueous solutions have at body temperature a low viscosity when equilibrated in the dark and undergo a sol-gel transition when irradiated. Such a transition strongly alters the diffusion of solutes such as methylene blue within the solutions. This light-induced interaction between the azobenzene moieties of DMA-MOAB and F127 micelles disappears when hydroxypropyl-beta-cyclodextrin (HPbetaCD) is added to the medium. In the presence of HPbetaCD, the cis-azobenzene groups are hosted in the cyclodextrin cavities and the mixed micelles are not formed. Therefore, changes in HPbetaCD concentration could be used to modulate the response of the copolymer blends to light. PMID- 17918872 TI - "Fingertip"-guided noncovalent functionalization of carbon nanotubes by dendrons. AB - Noncovalent functionalization of carbon nanotubes (CNTs) by dendrons was demonstrated. Certain types of dendrons successfully functionalized CNT surfaces through the noncovalent interactions between the peripheries of the dendrons and the sidewalls of CNTs. Dendrons have a unique anisotropic shape and an orthogonal functional group at their apex, and thus can generate a certain spacing between the functional groups upon immobilization on surfaces. Atomic force microscope (AFM) imaging, dispersion experiments, and MicroRaman spectroscopy were employed for the characterization of the functionalization. The binding was found to be governed by the chemical nature of the terminal groups, namely, the "fingertips", through a comparison study on the adsorption efficiency of the dendron analogs. Functional groups such as the carboxylic acid group and the benzyl amide group were effective for the cooperative binding. AFM analysis showed that the average spacing generated by the dendrons was 14-15 nm at a particular adsorption condition. Assembling streptavidin on the tubes through the dendrons and biotin confirmed the realization of the regulated spacing as well as the elimination of unwanted aggregation. The noncovalent functionalization of CNTs by a dendron can be a new approach toward sensible nanobiodevices, not only by introducing biomolecular probes on CNTs without disruption of the electronic network of the tubes, but also by providing the immobilized probe molecules with a space ample enough to minimize steric hindrance for the unhindered interaction with their target species. PMID- 17918873 TI - Vaporization-condensation-recrystallization process-mediated synthesis of helical m-aminobenzoic acid nanobelts. AB - One-dimensional (1D) helical organic nanostructures were synthesized by a modified vapor-solid (VS) process, called the vaporization-condensation recrystallization (VCR) process. The conventional solution-phase synthetic methods generally mediate self-assemblies of repeating unit molecules. To provide enough intermolecular interaction forces among the unit molecules, such strategy requires specific designs and syntheses of complex unit molecules as they possess numerous functional groups including phenyl rings, hydroxyl groups, long aliphatic chains, etc. On the contrary, we found that small and simple organic molecules, for example, m-ABA, could be self-assembled by the VCR process, resulting in 1D helical organic nanostructures. When m-aminobenzoic acid (m-ABA) powders were vaporized and transported to be condensed on a cooler region, the condensates were recrystallized into 1D helical nanobelts. Each step of the VCR process was confirmed from control experiments performed by varying reaction times, substrate types, and reaction temperatures. Powder XRD data, SAED analysis, and theoretical calculations revealed that dimers of m-ABA molecules have repeating units, and the growth axis of m-ABA nanohelices is [100]. PMID- 17918874 TI - Optimization of MALDI-TOF MS detection for enhanced sensitivity of affinity captured proteins spanning a 100 kDa mass range. AB - Analysis of complex biological samples by MALDI-TOF mass spectrometry has been generally limited to the detection of low-mass protein (or protein fragment) peaks. We have extended the mass range of MALDI-TOF high-sensitivity detection by an order of magnitude through the combined optimization of instrument parameters, data processing, and sample preparation procedures for affinity capture. WCX, C3, and IMAC magnetic beads were determined to be complementary and most favorable for broad mass range protein profiling. Key instrument parameters for extending mass range included adjustment of the ADC offset and preamplifier filter values of the TOF detector. Data processing was improved by a combination of constant and quadratic down-sampling, preceded by exponential baseline subtraction, to increase sensitivity of signal peaks. This enhancement in broad mass range detection of protein signals will be of direct benefit in MS expression profiling studies requiring full linear range mass detection. PMID- 17918875 TI - A new approach for mapping sialylated N-glycosites in serum proteomes. AB - A new approach for proteome-wide analysis of sialylated N-glycopeptides based on the diagonal chromatographic COFRADIC technology is presented here. The use of alpha(2-3,6,8,9) neuraminidase is central to isolate sialylated N-glycopeptides out of a complex peptide mixture. Two different COFRADIC techniques are introduced here, either without or with post-metabolic oxygen-18 labeling (direct versus indirect sorting), and when applied to immuno-depleted mouse serum, we herewith identified 93 sialylated glycosylation sites in 53 serum proteins. PMID- 17918876 TI - Direct counting of submicrometer-sized photosynthetic apparatus dispersed in medium at cryogenic temperature by confocal laser fluorescence microscopy: estimation of the number of bacteriochlorophyll c in single light-harvesting antenna complexes chlorosomes of green photosynthetic bacteria. AB - The number of pigments in single light-harvesting complexes (chlorosomes) were calculated by imaging single chlorosomes in a frozen buffer at cryogenic temperature with a confocal laser fluorescence microscope and pigment extraction. Chlorosomes were isolated from two types of green photosynthetic bacteria Chlorobium (Chl.) tepidum and Chloroflexus (Cfl.) aurantiacus and were individually imaged in the frozen medium. Each fluorescence spot observed mainly came from a single chlorosome and was ascribable to self-aggregates of bacteriochlorophyll (BChl) c molecules as core parts of chlorosomes. A three dimensional distribution of fluorescence of single chlorosomes was analyzed, and the number of chlorosomes in a volume of 54,000 microm(3) was counted directly. On the basis of the results, averaged numbers of the BChl c molecules contained in a single chlorosome of Chl. tepidum and Cfl. aurantiacus were determined to be 1.4 x 10(5) and 9.6 x 10(4), respectively. The present numbers are almost comparable to those estimated by other methods (Martinez-Planells et al., Photosynth. Res. 2002, 71, 83 and Montano et al., Biophys. J. 2003, 85, 2560). PMID- 17918877 TI - Aggregation of POSS monomers in liquid hexane: a molecular-simulation study. AB - Polyhedral oligomeric silsesquioxanes (POSS) are multifunctional molecules that can be employed as building blocks to develop nanocomposite materials whose mechanical properties often improve upon those of traditional polymeric materials. We report here molecular simulation results for the effective potential of mean force between octamethyl POSS monomers and between POSS monomers in which one methyl group has been substituted by a linear alkane chain of nine carbon atoms in liquid normal hexane at 300 and 400 K. The results are discussed and compared to available data for the effective interactions between octamethyl POSS monomers in normal hexadecane. Our results show that the effective short-ranged POSS-POSS attraction is significantly weaker in hexane than it is in hexadecane, perhaps explaining why normal hexane is often the solvent of choice for the preparation of POSS-containing materials. Additionally, we provide results for the radial distribution functions between selected sites in the POSS monomers that can be used both to understand the association between POSS monomers in solution and to parametrize coarse-grained simulation models. Such models will be used to study the formation of POSS-containing supramolecular structures such as lamellae or micelles that are currently not accessible by atomistic simulation and can be compared to experimental observations. PMID- 17918878 TI - Raman observation of the interactions between NH4+, SO4(2-), and H2O in supersaturated (NH4)2SO4 droplets. AB - High signal-to-noise ratio (S/N) Raman spectra of (NH(4))(2)SO(4) droplets deposited on a quartz substrate were obtained from dilute to supersaturated states upon decreasing the relative humidity (RH). When the molar water-to-solute ratio (WSR) decreases from 16.8 to 3.2, the v(1)-SO(4)(2-) band changes very little, that is, showing a red-shift of only about 1 cm(-1) (from 979.9 to 978.8 cm(-1)) and an increase of its full width at half-maximum (fwhm) from 8.3 to 9.8 cm(-1). Other vibration modes such as v(2)- and v(4)-SO(4)(2-) bands appear almost constantly at 452 and 615 cm(-1). Such kind of a spectroscopic characteristic is different from previous observation on other cations, indicating that the interactions between SO(4)(2-) and NH(4)+ in supersaturated states are similar to those between SO(4)(2-) and H(2)O in dilute states. After fitting the Raman spectra with Gaussian functions in the spectral range of 2400 4000 cm(-1), we successfully extracted six components at positions of 2878.7, 3032.1, 3115.0, 3248.9, 3468.4, and 3628.8 cm(-1), respectively. The first three components are assigned to the second overtone of NH(4)+ umbrella bending, the combination band of NH(4)+ umbrella bending and rocking vibrations, and the NH(4)+ symmetric stretching vibration, while the latter three components are from the strongly, weakly, and slightly hydrogen-bonded components of water molecules, respectively. With a decrease of the RH, the proportion of the strongly hydrogen bonded components increases, while that of the weakly hydrogen-bonded components decreases in the droplets. The coexistence of strongly, weakly, and slightly hydrogen-bonded water molecules must hint at a similar hydrogen-bonding network of NH(4)+, SO(4)(2-), and H(2)O to that of pure liquid water in supersaturated (NH(4))(2)SO(4) droplets. PMID- 17918879 TI - Hamiltonian and distance replica exchange method studies of Met-enkephalin. AB - The conformational states of the zwitterionic form of the pentapeptide Met enkephalin were explored with the use of explicit solvent molecular dynamics (MD). The N and C termini are ionized, as appropriate to polar solvent conditions, and consequently, there is a competition between open forms driven by polar solvation of the ammonium and carboxylate groups and closed forms driven by their salt-bridge formation. Normal MD started from an open state does not sample closed conformations. Sampling was enhanced with a distance replica exchange method (DREM) and with a Hamiltonian replica exchange method (HREM). The potential of mean force (PMF) along an end-to-end distance reaction coordinate was obtained with the DREM. The PMF shows a stable salt-bridge state and the presence of a large region of open states, as hypothesized for conformationally promiscuous small opiate peptides. The HREM systems differ by scaling the peptide peptide and peptide-solvent electrostatic and Lennard-Jones potentials, with the goal of improving the sampling efficiency with a limited number of systems. A small number of systems were found to be sufficient to sample closed and open states. A principal component analysis (PCA) shows that the HREM-generated fluctuations are dominated by the first two principal modes. The first corresponds to the end-to-end reaction coordinate found in the DREM, and the first mode PMF is similar to the DREM PMF. The second mode describes the presence of two conformations, both of which correspond to the salt-bridge state distance. The conformers differ in the values of neighboring psi and phi dihedral angles, since such psi/phi compensation can still produce the same end-to-end distance. The two-dimensional PMF constructed from the first two PCA modes captures most of the significant backbone conformational space of Met-enkephalin. PMID- 17918880 TI - Implicit nonpolar solvent models. AB - We have systematically analyzed a new nonpolar solvent model that separates nonpolar solvation free energy into repulsive and attractive components. Our analysis shows that either molecular surfaces or volumes can be used to correlate with repulsive free energies of tested molecules in explicit solvent with correlation coefficients higher than 0.99. In addition, the attractive free energies in explicit solvent can also be reproduced with the new model with a correlation coefficient higher than 0.999. Given each component optimized, the new nonpolar solvent model is found to reproduce monomer nonpolar solvation free energies in explicit solvent very well. However, the overall accuracy of the nonpolar solvation free energies is lower than that of each component. In the more challenging dimer test cases, the agreement of the new model with explicit solvent is less impressive. Nevertheless, it is found that the new model works reasonably well for reproducing the relative nonpolar free energy landscapes near the global minimum of the dimer complexes. PMID- 17918881 TI - Pi-dimer of an aniline dimer: an ESR-UV-vis spectroelectrochemical study. AB - It is shown for the first time that the most important intermediate formed during aniline polymerization, the p-aminodiphenylamine, forms a pi-dimer under oxidation at room temperature in acidified organic solvents that are used in electropolymerization. N-Phenylquinonediimine, which is generally assumed to be formed under oxidation, is only formed in basic solutions and in ionic liquids. Most of the mechanistic studies reported so far take the formation of N phenylquinonediimine under consideration, although it is not consistent with the UV-vis spectra measured during oxidation of p-aminodiphenylamine. The formation of a pi-dimer is very well consistent with the electronic spectra of the oxidation product. In this way the pi-dimer is very important for the interpretation of the UV-vis spectra of higher oligomers and polyaniline as well. Furthermore, it offers a new interpretation of the redox behavior of p aminodiphenylamine as found by cyclic voltammetry and has to be considered in the mechanism of the electrochemical polyaniline formation. PMID- 17918883 TI - Symmetry numbers and statistical factors in self-assembly and multivalency. AB - Evaluation of statistical factors in self-assembly processes is not a firmly settled question. As a contribution to solve this problem, a critical re examination of the symmetry number method and generalization of the direct count method are presented. The two approaches, producing the same results, mutually reinforce their role with respect to other discordant methods whose results cannot be independently checked. The direct count method moreover serves as a rationale for the apparently odd results the symmetry number method sometimes provides. The two methods thus turn out to be complementary to each other. Discussion of some exemplary cases points to the importance and subtlety of the role played by the geometrical features of assemblies involving intramolecular bonds. PMID- 17918882 TI - Modified calix[4]pyrrole receptor: solution thermodynamics of anion complexation and a preliminary account on the phosphate extraction ability of its oligomer. AB - A modified calix[4]pyrrole, namely meso-tetramethyl-tetrakis-(4-hydroxyphenyl) calix[4]pyrrole, 1, has been synthesized and characterized. (1)H NMR investigations in various deuterated solvents seems to indicate that this receptor interacts with acetone-d(6). The solution thermodynamics of 1 in various solvents is reported. Complexation studies in CD(3)CN show that the NH and OH functionalities of 1 are the active sites of its interaction with the fluoride and the dihydrogen phosphate anions. The composition of the anion complexes was established through conductance measurements. In all cases, 1:1 complexes are formed. The thermodynamics of anion complexation in acetonitrile and N,N dimethylformamide is discussed comparatively with previous reported data for the parent calix[4]pyrrole, 2, and these anions in these solvents. The medium effect on anion complexation is discussed in terms of the solvation properties of the reactants and the product in acetonitrile and N,N-dimethylformamide. An oligomeric material containing 1 as anchor group was synthesized and characterized by mass spectrometry. Preliminary studies have been performed to assess the extracting properties of this oligomer for the removal of phosphates from aqueous solutions. The effects of pH, temperature on the extraction of this anion salt from water, as well as the kinetics of the process (fast) were investigated. PMID- 17918884 TI - Rotation and diffusion of H2 in hydrogen-ice clathrate by 1H NMR. AB - A recently reported hydrogen-ice clathrate carries up to four H(2) in each large cage and one H(2) in each small cage. We report pulsed proton NMR line shape measurements on H(2)-D(2)O clathrate formed at 1500 bar and 250 K. The behavior of the two-pulse spin-echo amplitude with respect to the nutation angle of the refocusing pulse shows that intramolecular dipolar broadening, modulated by H(2) molecular reorientations, dominates the line width of the ortho-H(2). Dipolar interaction between H(2) guests and host D atoms explains the echo variation with the relative phases of the pulses. From 12 to 120 K, the line width varies as 1/T, demonstrating that the three sublevels of J = 1 are split by a constant energy, epsilon. The splitting arises from distortion in the otherwise high symmetry cages from frozen-out D(2)O orientational disorder. Above 120 K, further line-narrowing signals the onset of H(2) diffusion from cage to cage. At the lowest temperature, 1.9 K, the spectrum has Pake powder doublet-like features; the doublet is not fully developed, indicating a broad distribution of order parameters and energies epsilon. PMID- 17918885 TI - Effect of ionic surfactants on the hydration behavior of triblock copolymer micelles: a solvation dynamics study of coumarin 153. AB - Dynamic fluorescence Stokes shift measurements of coumarin 153 (C153) have been carried out to study the influence of ionic surfactants (sodium dodecyl sulfate, SDS and hexadecyltrimethylammonium chloride, CTAC) on the hydration behavior of aqueous poly(ethylene oxide)(20)-poly(propylene oxide)(70)-poly(ethylene oxide)20 (P123) block copolymer micelles. Increase in SDS or CTAC concentration at a fixed P123 concentration induces the steady-state emission spectra of C153 to shift gradually toward lower energy. This is attributed to an increase in polarity (due to enhanced hydration) experienced by the probe as a consequence of incorporation of ionic head groups in the Corona region. The observed dynamic fluorescence Stokes shift value decreases more in mixed micellar systems than in pure copolymer micelles and the trends are quite similar in the presence of SDS and CTAC. The spectral shift correlation functions were observed to be nonexponential in nature. Critical analysis of the spectral shift correlation function indicates a fast solvation component (<0.2 ns) in P123 micelles, which was absent in the presence of ionic surfactants. Due to increased hydration in the presence of ionic surfactants, the initial fast solvation event was elusive in mixed copolymer-surfactant systems, reflecting the absence of faster solvation component and reduced observed Stokes shift in mixed systems. It has been argued that in the low surfactant concentration region, increase in hydration with the incorporation of ionic head groups in the Corona region is mainly due to increase in mechanically trapped water content. However, at higher surfactant concentrations, bound water content dominates and leads to slower solvation dynamics. The present results also indicate that though CTAC alters the Corona hydration more efficiently than SDS, the overall influence of ionic surfactants on the Corona hydration is grossly similar irrespective of the cationic or anionic nature of the surfactants. Interaction of SDS and CTAC with poly(ethylene oxide)(100)-poly(propylene oxide)(70)-poly(ethylene oxide)(100) (F127) block copolymer micelles has also been studied to comprehend the effect of copolymer composition. The overall trends in dynamic fluorescence Stokes shift and solvation times are similar in both the copolymer micelles. PMID- 17918886 TI - Generalized phase behavior of small molecules and nanoparticles. AB - Prediction and understanding of the thermodynamic properties and kinetics of phase transitions in molecular systems depends on tuning intermolecular interactions such that the desired structures are assembled. These interactions can depend on the solvent temperature and composition and are difficult to determine in an a priori manner. This is especially true for large and complex molecules and nanoparticles with functionalized surfaces. Here, we demonstrate the use of the pair contribution of the long-time self-diffusivity determined by pulsed-field gradient spin-echo nuclear magnetic resonance as a probe of these interactions. Materials with high solubilities have scaled long-time self diffusivity, D2, values that are close to hard sphere values and decrease as the solubility decreases. We find a remarkable correlation between solubility and D2 for a wide range of hydrogen-bonding solutes that crystallize upon quenching solutions from high temperature. This generalized phase behavior can be understood in terms of the solutes' interacting with attractive forces that have an extent that is only a small fraction of their diameters. PMID- 17918887 TI - Reversible redox processes of poly(anilines) in layered semiconductor niobate films under alternate UV-vis light illumination. AB - The synthesis of polyaniline (PANI) with semiconducting layered niobate (NbO) to form PANI/NbO hybrid materials and their reversible color change under a unique redox process under the influence of UV and/or visible light have been investigated. The in-situ polymerization of anilinium chlorides (ANI) packed in a regular orientation in a bilayer structure within the NbO interlayers led to PANI/NbO hybrid powders by heat treatment using (NH(4))(2)S(2)O(8) as the catalyst. The resulting PANI of these hybrids showed the characteristics of a fully oxidized quinoid form, i.e., pernigranine (PG). The PANI/NbO suspension in H(2)O was cast on a glass substrate to form a PANI/NbO film after evaporation of the water with a good parallel orientation of the NbO layers against the glass substrate. Upon UV light irradiation in the presence of a reductant such as MeOH, the violet-colored PANI (PG) polymers within the NbO interlayers were reduced by the NbO-induced photocatalytic reactions and led to a colorless PANI, i.e., leucoemeraldine (LE). Moreover, the resulting colorless PANI/NbO films reverted back to a blue-colored PANI, i.e., emeraldine (EM), due to oxidation by the surrounding O(2) gas. The PANI/NbO hybrid films were able to retain repetitive and reversible photoinduced patterning for over 50 cycles under such alternate UV and visible light irradiation. PMID- 17918889 TI - Direct conjugation of semiconductor nanocrystals to a globular protein to study protein-folding intermediates. AB - In this Article, we study the development of semiconductor nanocrystals (quantum dots of average diameter less than 2 nm) directly conjugated to a transporter protein human serum albumin (HSA) as fluorescent biological labels. Forster resonance energy transfer (FRET) from the amino acid tryptophan (Trp214) to quantum dot in HSA is monitored to follow the local and global changes in the protein structure during thermal unfolding and refolding processes. This study is likely to attract widespread attention as a powerful tool for the study of protein folding. PMID- 17918888 TI - Probing the micellization kinetics of pyrene end-labeled diblock copolymer via a combination of stopped-flow light-scattering and fluorescence techniques. AB - A pyrene end-labeled double hydrophilic diblock copolymer, poly(2 (diethylamino)ethyl methacrylate)-b-poly(2-(dimethylamino)ethyl methacrylate) (Py PDEA-b-PDMA), was synthesized by sequential monomer addition via oxyanionic polymerization using a 1-pyrenemethanol-based initiator. This diblock copolymer exhibits reversible pH-responsive micellization behavior in aqueous solution, forming PDEA-core micelles stabilized by the soluble PDMA block at neutral or alkaline pH. Taking advantage of the pyrene probe covalently attached to the end of the PDEA block, the pH-induced micellization kinetics of Py-PDEA-b-PDMA was monitored by stopped-flow light scattering using a fluorescence detector. Upon a pH jump from 4.0 to 9.0, both the scattered light intensity and excimer/monomer fluorescence intensity ratios (IE/IM) increase abruptly initially, followed by a more gradual increase to reach plateau values. Interestingly, the IE/IM ratio increases abruptly within the first 10 ms: a triple exponential function is needed to fit the corresponding dynamic trace, leading to three characteristic relaxation time constants (tau(1,fluo) < tau(2,fluo) < tau(3,fluo)). On the other hand, dynamic traces for the scattered light intensity can be well-fitted by double exponential functions: the resulting time constants tau(1,scat) and tau(2,scat) can be ascribed to formation of the quasi-equilibrium micelles and relaxation into their final equilibrium state, respectively. Most importantly, tau(1,scat) obtained from stopped-flow light scattering is in general agreement with tau(2,fluo) obtained from stopped-flow fluorescence. The fastest process (tau(1,fluo) approximately 4 ms) detected by stopped-flow fluorescence is ascribed to the burst formation of small transient micelles comprising only a few chains, which are too small to be detected by conventional light scattering. These nascent micelles undergo rapid fusion and grow into quasi-equilibrium micelles and then slowly approach their final equilibrium state. The latter two processes can be detected by both techniques. PMID- 17918890 TI - An H/D isotopic substitution study of the H5O2+.Ar vibrational predissociation spectra: exploring the putative role of Fermi resonances in the bridging proton fundamentals. AB - To clarify the nature of the motions contributing to the observed multiplet structures in the low-energy (900-1800 cm-1) vibrational spectrum of the H5O2+ "Zundel" ion, we report the evolution of its vibrational fingerprint with sequential H/D isotopic substitution in a predissociation study of the Ar complexes. Of particular interest is the D4HO2+ complex, which displays a single intense band in the vicinity of the asymmetric OHO stretch of the bridging proton, in contrast to the more complex multiplet observed for both H5O2+ and D5O2+ isotopologues. These intensity patterns are consistent with the recent assignment of the bridging proton band's doublet in the H5O2+.Ne spectrum to a 2 x 2 Fermi resonance interaction between the shared proton stretch and a complex background level primarily derived from the O-O stretch together with two quanta of the wagging vibration involving the pyramidal deformations of the flanking H2O groups (Vendrell, O.; Gatti, F.; Meyer, H.-D. Angew. Chem., Int. Ed. 2007, 46, 6918). In addition, the observed trends rule out assignment of the approximately 1800 cm-1 feature in H5O2+ to a combination band of the bridging proton vibration with the O-O stretch, providing a secure foundation for the previously reported scheme that attributes this band to the out-of-phase intramolecular bending fundamental. The observed feature occurs at an unusually high energy for typical HOH bends, however, and we explore the participation of the bridging proton in these eigenstates by following how the calculated harmonic spectrum evolves when artificially large masses are assigned to the proton. The empirical assignments are supported by anharmonic estimates of the isotope shifts evaluated by the diffusion Monte Carlo method. PMID- 17918891 TI - Polymorph selection during the crystallization of softly repulsive spheres: the inverse power law potential. AB - Using hybrid Monte Carlo molecular simulations, we study crystallization from the melt of softly repulsive spheres interacting through an inverse power law potential. We work at fixed supercooling (i.e., at a temperature 25% below the melting temperature) and consider three systems, defined by different values for the inverse power exponent n: n = 5, n = 6.67, and n = 10. Modifying the value of n allows us to study the onset of crystallization in the domain of stability of the body-centered cubic (bcc) phase (n = 5 and n = 6.67) and in the domain of stability of the face-centered cubic (fcc) phase (n = 10). We show that, for the three systems, polymorph selection does not take place during crystal nucleation since the structure of the critical nuclei obtained for the three systems is not well defined. However, our results demonstrate that polymorph selection takes place during the growth step since growth proceeds either into the stable bcc phase for the two smaller values of n (n = 5 and n = 6.67) or into the stable fcc phase for the larger value of n (n = 10). We also show that we did not achieve complete control of polymorphism for n = 10. The growth step gives rise to either slowly growing crystallites composed of two blocks of different structures (the stable fcc form and the metastable bcc form) or rapidly growing crystallites of the metastable bcc form. PMID- 17918892 TI - Triplet-polaron quenching in conjugated polymers. AB - We studied the triplet-polaron quenching in a platinum(II) porphyrin- (PtOEP-) doped polyspirobifluorene (PSF-TAD) copolymer. The copolymer contains a hole transporting phenylenediamine unit (TAD) as a comonomer. Triplet-polaron quenching was probed by the change in PtOEP phosphorescence lifetime under an applied voltage in a unipolar device. The charge-induced reduction of the optically excited lifetime of PtOEP is one-third for the highest applied bias. The charge density can be obtained from current-voltage characteristics in the space-charge-limited (SCL) regime. The obtained hole mobility under SCL conditions is (7 +/- 2) x 10(-5) cm(2)/(V s). This result is in accord with recent mobility measurements of the time-of-flight mobility in our polymer. The triplet-polaron recombination constant was evaluated to be (4 +/- 1) x 10(-13) cm(3)/s, implying a triplet-polaron interaction radius of 2 x 10(-10) m. The results show that triplet-polaron annihilation cannot be neglected in device models for phosphorescent light-emitting diodes. PMID- 17918893 TI - Coexistence of homochiral and heterochiral adenine domains at the liquid/solid interface. AB - In this work, the self-assembly of the DNA base molecule adenine (A) is imaged with high-resolution scanning tunneling microscopy (STM) at the liquid (1 octanol)/solid (HOPG) interface at room temperature. Rather surprisingly, the STM results reveal, for the first time, the spontaneous formation of two coexisting distinct (homo- and heterochiral) domains of adenine, which are formed at the liquid/solid interface without changing any experimental conditions. Ab initio density functional theory (DFT) calculations support our STM findings and suggest the existence of various A networks of nearly similar stability that all are constructed from the most stable A dimer. PMID- 17918894 TI - Guanidino- and urea-modified dendrimers as potent solubilizers of misfolded prion protein aggregates under non-cytotoxic conditions. dependence on dendrimer generation and surface charge. AB - Amino-terminated dendrimers are well-defined synthetic hyperbranched polymers and have previously been shown to destabilize aggregates of the misfolded, pathogenic, and partially protease-resistant form of the prion protein (PrPSc), transforming it into a partially dissociated, protease-sensitive form with strongly reduced infectivity. The mechanism behind this is not known, but a low pH, creating multiple positively charged primary amines on the dendrimer surface, increases the efficiency of the reaction. In the present study, surface amines of the dendrimers were modified to yield either guanidino surface groups (being positively charged at neutral pH) or urea groups (uncharged). The ability of several generations of modified dendrimers and unmodified amino-terminated dendrimers to deplete PrPSc from persistently PrPSc-infected cells in culture (SMB cells) was studied. It was found that destabilization correlated with both the generation number of the dendrimer, with higher generations being more efficient, and the charge density of the surface groups. Urea-decorated dendrimers having an uncharged surface were less efficient than positively charged unmodified- (amino) and guanidino-modified dendrimers. The most efficient dendrimers (generation 4 (G4) and G5-unmodified and guanidino dendrimers) cleared PrPSc completely by incubation for 4 days at less than 50 nM. In contrast to both unmodified and guanidine-modified dendrimers, the uncharged urea dendrimers showed much lower cytotoxicity toward noninfected SMB cells. Therapeutic uses of modified dendrimers are indicated by the low concentrations of dendrimers needed. PMID- 17918895 TI - Synthesis and characterization of thermo- and pH-responsive double-hydrophilic diblock copolypeptides. AB - Synthesis of novel double-hydrophilic diblock copolypeptides (BCPs), poly(l glutamic acid)-block-poly(N-isopropylacrylamide) (PLGnPNm), and their thermoresponsive properties in aqueous solutions at different pH values are described. The diblock copolypeptides were synthesized by a combination of ring opening polymerization (ROP) of gamma-benzyl-l-glutamate N-carboxyanhydrides (BLG NCA) and reversible addition-fragmentation chain transfer (RAFT) polymerization of N-isopropylacrylamide (NiPAM). A new class of RAFT agents (CTA-2 and CTA-3) with amino-functional groups was designed for this purpose. Two different strategies, i.e., macrochain transfer agent (CTA) and macroinitiator routes, were utilized and compared on the control of the chemical structures of the resulting BCPs. Their block ratios and lengths are broadly varied (n = 21-600 and m =180 442). Their thermally switchable aggregation behaviors in aqueous solutions were investigated at the microscopic level by 1H NMR spectroscopy and at the macroscopic level by turbidity measurements using UV/vis spectroscopy. The latter was also utilized for their lower critical aggregation temperature (LCAT) determination. The effects of block lengths and ratios as well as solution pH values on the collapse of NiPAM chain and aggregation process of BCPs were examined. This aggregation process was also followed by dynamic light scattering (DLS) measurements, and the thermally induced aggregate structures were investigated by transmission electron microscopy (TEM). PMID- 17918896 TI - An intramolecular Diels-Alder strategy for the asbestinins: enantioselective total syntheses of 11-acetoxy-4-deoxyasbestinin D and asbestinin-12. AB - The enantioselective total syntheses of 11-acetoxy-4-deoxyasbestinin D and asbestinin-12 have been completed. A glycolate aldol reaction provided a diene useful for ring-closing metathesis to form an oxonene, which was ultimately employed as a template to execute a highly stereoselective intramolecular Diels Alder cycloaddition, forming the hydroisobenzofuran moiety. The absolute configuration of the asbestinin subclass was confirmed via these synthetic efforts. PMID- 17918897 TI - A novel strategy to assemble the beta-diketo acid pharmacophore of HIV integrase inhibitors on purine nucleobase scaffolds. AB - Claisen condensation, the key step in constructing the pharmacophore of aryl beta diketo acids (DKA) as integrase inhibitors, fails in certain cases of highly electron-deficient heterocycles such as purines. A general synthetic strategy to assemble the DKA motif on the purine scaffold has been accomplished. The synthetic sequence entails a palladium-catalyzed cross-coupling, a C-acylation involving a tandem addition/elimination reaction, and a novel ferric ion catalyzed selective hydrolysis of an enolic ether in the presence of a carboxylic acid ester. PMID- 17918898 TI - Syntheses and reactivity of meso-unsubstituted azuliporphyrins derived from 6 tert-butyl- and 6-phenylazulene. AB - A series of six azuliporphyrins with substituents on the seven-membered ring were prepared by two different "3 + 1" routes from 6-tert-butyl- and 6-phenylazulene. The substituted azulenes can be converted into dialdehydes under Vilsmeier-Haack conditions, and these react with tripyrranes in the presence of TFA in CH2Cl2 to give azuliporphyrins in excellent yields. Alternatively, tripyrrane analogues can be prepared by reacting the substituted azulenes with an acetoxymethylpyrrole in the presence of acetic acid, and following a deprotection step, these condensed with a pyrrole dialdehyde to give the related azuliporphyrins in 45-51% yield. Five of the azuliporphyrins were sufficiently soluble in CDCl3 to afford high quality proton and carbon-13 NMR data. The internal CH and NH resonances were observed near 3 ppm, although the precise values were dependent upon substituent effects. The presence of a tert-butyl group on the azulene moiety slightly enhanced the diatropicity of the macrocycle compared to the phenyl-substituted azuliporphyrins. Polar solvents also increased the downfield shifts to the external protons by stabilizing the dipolar resonance contributors that are responsible for the carbaporphyrinoid aromatic character. A tert-butyl substituted azuliporphyrin also gave X-ray quality crystals, and this allowed the first structural analysis of a free base azuliporphyrin to be conducted. The macrocycle is near planar, and the azulene unit was only tilted out of the plane by 7.4 degrees. An analysis of the bond lengths suggests that a 17 atom delocalization pathway significantly contributes to the aromatic properties of this system. Protonation of azuliporphyrins affords dications with enhanced diamagnetic ring currents where the internal CH shifts to ca. -3 ppm. Again, the chemical shifts are influenced by the substituents and the presence of an electron-donating tert-butyl group on the azulene subunit increases the macrocyclic diatropicity. Two of the substituted azuliporphyrins were reacted with nickel(II) acetate or palladium(II) acetate in DMF to give the corresponding organometallic derivatives, and these stable complexes were isolated in excellent yields. Addition of pyrrolidine to NMR solutions of 23-substituted azuliporphyrins 19 demonstrated that nucleophilic addition products were present in equilibrium with the parent porphyrinoids, but these adducts are less favored than for azuliporphyrins lacking the 23-substituents. Although nucleophilic attack of a peroxide anion is believed to be the first step in the conversion of azuliporphyrins to benzocarbaporphyrins with t-BuOOH and KOH, the tert-butyl or phenyl substituents in azuliporphyrins 19a and 19b did not inhibit this chemistry. Two benzocarbaporphyrin products were isolated and characterized in each case, and mechanisms are proposed to explain the origins of these oxidative ring contraction products. PMID- 17918899 TI - New light on an old story: facile and efficient synthesis of 1,3-diaryl-5 spirohexahydropyrimidines via a six-molecule, three-component mannich-type reaction. AB - A facile and efficient synthesis of 1,3-diaryl-5-spirohexahydropyrimidines via a one-pot condensation of anilines, formaldehyde, and cyclohexanones is reported. In this one-pot, three-component reaction, six molecules of reactants are involved and six new covalent bonds are generated. Bicyclic products are obtained from the starting materials in one pot using readily available starting materials and catalysts. PMID- 17918900 TI - Dramatic effects of ionic liquids on the palladium-catalyzed cyclocarbonylation of enynols with thiols. AB - Palladium-catalyzed carbonylation reactions of enynols with thiols in ionic liquids afford monocarbonylated 6-membered-ring lactones in high yields and good selectivity. These results are significantly different from those obtained when conducting the reaction in THF. The recyclability of the catalytic system was also investigated. PMID- 17918901 TI - Disubstituted pyridines: the double-coupling approach. AB - A one-pot procedure leading to disubstituted pyridines from the starting dibromopyridines is described. Key features include the ability to couple a range of aryl and even alkenylboronic acids at the 2,3 and/or 2,5 positions with excellent regiocontrol under a standard set of conditions. Further, isolated yields are greatly improved by the use of neutral alumina in place of silica for product purification. Finally, the intrinsic electronic bias of the pyridine ring can be overcome by using a bromoiodopyridine. PMID- 17918902 TI - 9-phenyl-10-methylacridinium: a highly efficient and reusable organocatalyst for mild aromatization of 1,4-dihydropyridines by molecular oxygen. AB - The mild, efficient, and general aerobic aromatization of Hantzsch 1,4 dihydropyridines was realized at room temperature with 5 mol % of 9-phenyl-10 methylacridinium perchlorate as the photocatalyst, which could be easily recovered and reused. PMID- 17918903 TI - Active nanodiamond hydrogels for chemotherapeutic delivery. AB - Nanodiamond materials can serve as highly versatile platforms for the controlled functionalization and delivery of a wide spectrum of therapeutic elements. In this work, doxorubicin hydrochloride (DOX), an apoptosis-inducing drug widely used in chemotherapy, was successfully applied toward the functionalization of nanodiamond materials (NDs, 2-8 nm) and introduced toward murine macrophages as well as human colorectal carcinoma cells with preserved efficacy. The adsorption of DOX onto the NDs and its reversible release were achieved by regulating Cl- ion concentration, and the NDs were found to be able to efficiently ferry the drug inside living cells. Comprehensive bioassays were performed to assess and confirm the innate biocompatibility of the NDs, via real-time quantitative polymerase chain reaction (RT-PCR), and electrophoretic DNA fragmentation as well as MTT analysis confirmed the functional apoptosis-inducing mechanisms driven by the DOX-functionalized NDs. We extended the applicability of the DOX-ND composites toward a translational context, where MTT assays were performed on the HT-29 colon cancer cell line to assess DOX-ND induced cell death and ND-mediated chemotherapeutic sequestering for potential slow/sustained released capabilities. These and other medically relevant capabilities enabled by the NDs forge its strong potential as a therapeutically significant nanomaterial. PMID- 17918904 TI - Two-dimensional imaging by far-field superlens at visible wavelengths. AB - We report that two-dimensional (2D) sub-diffraction-limited images can be theoretically reconstructed by a new metamaterial far-field superlens. The metamaterial far-field superlens, composed of a metal-dielectric multilayer and a one-dimensional (1D) subwavelength grating, can work over a broad range of visible wavelengths intrinsically. The imaging principle and the reconstruction process are described in detail. The 2D sub-diffraction-limited imaging ability enables more applications of the far-field superlens in optical nanoimaging and sensing. PMID- 17918905 TI - Efficiency enhancements in solid-state hybrid solar cells via reduced charge recombination and increased light capture. AB - We compare a series of molecular sensitizers in dye-sensitized solar cells containing the organic hole transporter 2,2',7,7'-tetrakis(N,N-di-p-methoxypheny amine)-9,9'-spirobifluorene (spiro-MeOTAD). Charge recombination is reduced by the presence of "ion-coordinating" moieties on the dye, with the longest electron lifetime and highest solar cell efficiency achieved using a novel sensitizer with diblock alkoxy-alkane pendent groups. By further increasing the optical path length in the active layer, we achieve a power conversion efficiency of over 5% under simulated sun light. PMID- 17918906 TI - LC-MS/MS assay for protein amino acids and metabolically related compounds for large-scale screening of metabolic phenotypes. AB - A modified LC-MS/MS method for large-scale screening of metabolic phenotypes was developed and validated. Twenty amino acids and 5 metabolically related compounds are measured within 4 min using multiple reaction monitoring (MRM) transitions selective for each compound. Separation with a short C18 column and rapid gradient using the ion-pairing reagent perfluoroheptanoic acid allows chromatographic resolution of the isomers Ile and Leu and improved chromatographic peak shapes for Lys, Arg, and His. MRM transitions were established with capability to distinguish isomers Leu from Ile and Thr from homoserine even when chromatographic resolution is incomplete. The reproducibility of the assay was tested by adding eight stable isotope-labeled amino acid standards (AA*) to extracts of Arabidopsis thaliana seeds. In intra- and interday assay comparisons, mean coefficients of variation of the peak area ratios of AA/AA* were less than 5% for all but Gly/Gly-D2-15N1. Recoveries of these eight amino acids ranged from 62 to 94% and suppression of signal by the matrix were 31-65%. Dilution of seed extracts reduced ion suppression for early eluting amino acids but had minimal effects for those eluting later. The intra- and interday accuracies for these eight amino acids were 77-131 and 88-133% of nominal concentrations, respectively. PMID- 17918907 TI - Monitoring the interactions of tocopherol homologues with reversed-phase stationary HPLC phases by 1H suspended-state saturation transfer difference high resolution/magic angle spinning NMR spectroscopy. AB - The separation process in reversed-phase high-performance liquid chromatography employing C18 phases is mainly due to hydrophobic interactions. The separation of tocopherol isomers, exhibited by the C30 phases, however, is additionally driven by shape selectivity. This phenomenon is investigated by suspended-state nuclear magnetic resonance spectroscopy using the saturation transfer difference technique, which was originally introduced to study protein-ligand interactions. The interaction strength between beta-/gamma-tocopherol and three different stationary phases was estimated qualitatively. The nuclear magnetic resonance data are compared to chromatographic data, and a similar mode of interaction between the analytes and the stationary phases is elucidated. PMID- 17918908 TI - Rapid screening of anabolic steroids in urine by reactive desorption electrospray ionization. AB - Fast screening for anabolic steroids in whole urine is achieved by combining reactive desorption electrospray ionization (reactive DESI) and tandem mass spectrometry. Spray solutions containing hydroxylamine allow heterogeneous reactions of hydroxylamine with the carbonyl group of the steroids during the ionization process. Seven steroids, including a glycosteroid, were examined. The ion/molecule reaction adduct and the oxime formed via its dehydration were observed using reactive DESI; the protonated and sodiated forms of the ionized steroid were also observed both in reactive DESI and in DESI performed without the added hydroxylamine reagent. Paper, glass, and polytetrafluoroethylene were tested as sample substrates, but the glycosteroid was ionized intact without hydrolysis only from polytetrafluoroethylene. Limits of detection for the pure compounds were less than 1 ng, dynamic ranges were typically 2 orders of magnitude, and analysis times were just a few seconds. Concentration levels of ketosteroids in raw urine relevant to screening for sports doping (approximately 20 ng/mL) can be reached using a simple solid-phase microextraction (SPME) preconcentration step. Reactive DESI provided significant improvements in ionization efficiency of these steroids in raw undiluted urine as compared to conventional DESI; suppression effects due to the sample matrix were minimal and the urine matrix had no deleterious effect on steroid detection limits. Tandem mass spectrometry provided confirmation of analyte identification in this rapid screening process. PMID- 17918909 TI - Use of docking peptides to design modular substrates with high efficiency for mitogen-activated protein kinase extracellular signal-regulated kinase. AB - The mitogen-activated protein kinase extracellular regulated kinase (ERK) plays a key role in the regulation of cellular proliferation. Mutations in the ERK cascade occur in 30% of malignant tumors. Thus understanding how the kinase identifies its cognate substrates as well as monitoring the activity of ERK is central to cancer research and therapeutic development. ERK binds to its protein targets, both downstream substrates and upstream activators, via a binding site distinct from the catalytic site of ERK. The substrate sequences that bind, or dock, to these sites on ERK influence the efficiency of phosphorylation. For this reason, simple peptide substrates containing only phosphorylation sequences typically possess low efficiencies for ERK. Appending short docking peptides derived from full-length protein substrates and activators of ERK to a phosphorylation sequence increased the affinity of ERK for the phosphorylation sequence by as much as 200-fold while only slightly diminishing the maximal velocity of the reaction. The efficiency of the phosphorylation reaction was increased by up to 150-fold, while the specificity of the substrate for ERK was preserved. Simple modular peptide substrates, which can be easily tailored to possess high phosphorylation efficiencies, will enhance our understanding of the regulation of ERK and provide a tool for the development of new kinase assays. PMID- 17918910 TI - Cytotoxic lignans from the stem bark of Magnolia officinalis. AB - Three new lignans, 4'-methoxymagndialdehyde ( 1), 4'-methoxymagnaldehyde B ( 2), and 4'-methoxymagnaldehyde E ( 3), were isolated from hexane- and EtOAc-soluble fractions of the stem bark of Magnolia officinalis, together with eight known compounds ( 4- 11). The structures of compounds 1- 3 were determined on the basis of spectroscopic and physicochemical data analysis. Compounds 1- 11 were tested in vitro for their cytotoxic activity against the K562, HeLa, and A549 cancer cell lines. Among the compounds tested, compound 1 showed the most potent cytotoxic activity against these cancer cell lines, with IC50 values of 3.9, 1.5, and 3.7 microg/mL, respectively. PMID- 17918911 TI - Solution-phase parallel synthesis of diverse 1,5-benzodiazepin-2-ones. AB - A practical and efficient parallel method has been developed for the synthesis of 1,5-benzodiazepin-2-ones with a large variety of substituents at the 3-, 4-, 5-, 7-, and 8-positions using 1,5-difluoro-2,4-dinitrobenzene as the starting material. All the reactions involved here are highly effective in giving the desired products under mild conditions. PMID- 17918912 TI - Multistage and tandem mass spectrometry of glycosylated triterpenoid saponins isolated from Bacopa monnieri: comparison of the information content provided by different techniques. AB - Whereas all state-of-the-art techniques in mass spectrometry (MS) have been extensively applied to oligosaccharides derived from glycoproteins, less effort has been devoted to the analysis of smaller glycoconjugates. In the present study, the application of a variety of MS techniques for the analysis of two dammarane-type triterpenoid saponins isolated from B. monnieri is reported. The structural information provided by ESI-ion trap (IT)-, AP-MALDI-IT-, and MALDI IT/reflectron time-of-flight (RTOF)-MS, all utilizing low-energy collision induced dissociation (CID), and MALDI-TOF/RTOF-MS, facilitating postsource decay and high-energy CID analysis, was compared. The applied desorption/ionization technique does not influence the fragmentation of identical precursor ions in low energy CID. All three fragmentation techniques clearly yield the sequence and branching of the glycan moiety as well as the molecular mass of the intact aglycon. Cross-ring cleavage of the branching sugar, which gives some information about the sugar linkages, was mainly observed in low-energy CID. High-energy CID, on the other hand, yielded some additional diagnostic fragment ions from the aglycon moiety. Internal cleavage ions are formed by alternative mechanisms in high-energy CID and are assumed to be diagnostic for some linkages. However, none of the applied MS techniques facilitates the identification of those saponins that differ only by their aglycon moiety (i.e., jujubogenin or pseudojujubogenin). PMID- 17918913 TI - Molecularly imprinted-matrix solid-phase dispersion for selective extraction of five fluoroquinolones in eggs and tissue. AB - A novel highly selective sample cleanup procedure combining molecular imprinting and matrix solid-phase dispersion (MI-MSPD) was developed for the simultaneous isolation of ofloxacin, pefloxacin, norflorxacin, ciprofloxacin, and enrofloxacin in chicken eggs and swine tissues followed by high-performance liquid chromatography with fluorescence detection. The novel ofloxacin imprinted polymers synthesized in water-containing systems show high selectivity for the five fluoroquinolones in aqueous environment and the affinity can be easily adjusted by the pH of solution. Compared with conventional MSPD methods, using MIPs as selective MSPD sorbents, the five fluoroquinolones could be selectively extracted from a biological matrix and all matrix interferences were eliminated simultaneously. The average recoveries of the five fluoroquinolones were ranged from 85.7 to 104.6% for eggs and 86.8 to 102.7% for tissues with relative standard deviations of less than 7.0%. Detection limits for the identification of the five fluoroquinolones in eggs and tissues ranged from 0.05 to 0.09 ng/g. PMID- 17918914 TI - Extended X-ray absorption fine structure investigation of adsorption and separation phenomena of metal ions in organic resin. AB - Analytical technique using organic resins has already been well-developed, and its applications are employed in various fields; nevertheless, the chemical phenomena occurring inside the resin remain unclear for the most part. In the present study, we apply EXAFS spectroscopy to elucidate the adsorption and separation phenomena of metal ions by organic resin. That is, the chemical species of trivalent lanthanides (Ln(III)) adsorbed in a tertiary pyridine resin from hydrochloric acid and nitric acid solutions have been determined by EXAFS. The results in HCl solutions suggest that Ln(III) ions are partly dehydrated in the resin phase, enabling the pyridine groups of the resin and chloride ions to coordinate to the Ln(III) ions in their primary coordination sphere. On the other hand, Ln(III) ions are tightly coordinated by several nitrate ions in HNO3 solutions and they keep forming the nitrate complex even in the resin phase. The lighter Ln of Nd tends to form an anionic nitrate complex, [Nd(NO3)4.nH2O]-, in the resin phase, while the middle Ln of Sm exists as a cationic nitrate complex, [Sm(NO3)2.nH2O]+, for the most part. On the basis of these EXAFS results, the adsorption and separation mechanisms of the pyridine resin in HCl solutions are interpreted as the direct coordination of pyridine groups to metal ions, while the mechanisms in HNO3 solutions are mainly dominated by the anion-exchange reaction between the protonated pyridine groups and the anionic nitrate complexes of Ln(III). The obtained results demonstrate that the hydration of metal ions weakens, and instead, other complexations are enhanced in the resin phase. PMID- 17918915 TI - Molecular similarity based on atomic electrostatic potential. AB - We propose a new similarity measure operating in the space spanned by the potential values, evaluated at atoms constituting the benzene ring and the COOH group in para-substituted benzoic acids and at benzene ring atoms in monosubstituted benzenes. The similarity measures are equivalent to the Euclidean distance between points in that space. Only the distances between the potentials at corresponding atoms in different molecules are included. The distances for benzene rings were very similar, regardless of whether they were calculated in para-substituted acids or in monosubstituted benzenes. As reference reactions, dissociation of benzoic acids and nitration of monosubstituted benzenes have been used. The effects of reduction of dimensionality of the potential space on the comparison of similarity measures with the free energies of the reference reactions have been investigated. It became obvious that the potentials at individual atoms in molecules of the acids and monosubstituted benzenes are mutually correlated to a high degree. PMID- 17918916 TI - Enhanced aqueous photochemical reaction rates after freezing. AB - Sunlit snow/ice is known to play an important role in the processing of atmospheric species, including photochemical production of NO(x), HONO, molecular halogens, alkyl halides, and carbonyl compounds, among others. It has been shown that a liquid-like (quasi-liquid or disordered) layer exists on the surface of pure ice and that this quasi-liquid layer is also found on the surface of ambient snow crystals and ice at temperatures similar to polar conditions. However, it is unclear what role the liquid-like fractions present in and on frozen water play in potential photochemical reactions, particularly with regard to organic substrates. Here, we report a detailed study of enhanced rates of photochemical nucleophilic substitution of p-nitroanisole (PNA) with pyridine, a well characterized and commonly used actinometer system. Reaction rates were enhanced by a factor of up to approximately 40 when frozen at temperatures between 236 and 272 K. Reaction rates were dependent on temperature and solute concentration, both variables that control the nature of the liquid-like fraction in frozen water. The results obtained indicate that a major portion of the organic solutes is excluded to the liquid-like layer, significantly impacting the rate of the photochemical nucleophilic substitution reaction studied here. Also, the direct comparison of liquid-phase kinetics to reactions occurring in frozen water systems is drawn into question, indicating that a simple extrapolation of liquid phase mechanisms to snow/ice may not be valid for certain reactions. PMID- 17918917 TI - Vibrational spectra of cyclopentadienylphosphine: infrared and theoretical studies from DFT anharmonic potentials. AB - Both experimental and theoretical infrared investigations of cyclopentadienylphosphine (CpP) are reported. The infrared spectra (3500-500 cm( 1)) in the gas phase have been recorded at 0.5 cm(-1) resolution. Infrared absorptions bands of the two lowest stable conformers were observed and assigned. Average integrated intensities of isolated and overlapping vibrational bands were also determined experimentally. The vibrational frequencies of the CpP system and its P-dideuterated isotopologue have been calculated by means of density functional theory. The Becke exchange functional and Lee-Yang-Parr correlation functional method with a combination of the two basis sets, namely 6-31+G(d,p) and the correlation-consistent triple-zeta cc-pVTZ set of Dunning, were used. Hybrid B3LYP/B3LYP//cc-pVTZ/6-31+G(d,p) anharmonic frequencies of the fundamental, overtone, and combination transitions were calculated in the 3500 200 cm(-1) area with the use of a variational approach, implemented in the P_Anhar_v1.1 code, to assign the experimental data for each conformer. PMID- 17918918 TI - Systematically convergent correlation consistent basis sets for molecular core valence correlation effects: the third-row atoms gallium through krypton. AB - The family of correlation consistent polarized valence basis sets has been extended in order to account for core-core and core-valence correlation effects within the third-row, main group atoms gallium through krypton. Construction of the basis sets is similar to that of the atoms boron through argon, where either the difference between core-correlated and valence-only correlation energies were calculated via configuration interaction (CISD) computations on the ground electronic states of the atoms (named cc-pCVnZ) or the sets were optimized with respect to the core-valence correlation energy and a small weight of core-core correlation energy (cc-pwCVnZ). Due to the correlation of 3d orbitals, added shells of higher angular momentum exponents compared to the valence sets are necessary to describe the core region. The pattern of added core-correlating functions is (1s1p1d1f) for double-zeta, (2s2p2d2f1g) for triple-zeta, (3s3p3d3f2g1h) for quadruple-zeta, and (4s4p4d4f3g2h1i) for quintuple-zeta. Atomic and molecular results show good convergence to the CBS limit, with the cc pwCVnZ sets showing improved convergence compared to the cc-pCVnZ ones for molecular core-valence correlation effects. After testing the basis sets on the homonuclear diatomics Ga2-Kr2 with coupled cluster wave functions, it is concluded that a treatment of core-valence correlation effects is essential for high-accuracy ab initio investigations of third-row-containing molecules. Though the basis sets are optimal for 3s3p3d correlation, preliminary atomic and molecular results show the basis sets to be efficient with respect to 3d-only correlation, and these potentially could be used with 3d-only correlation for more qualitative studies on larger species. PMID- 17918919 TI - Investigation of the amino acids glycine, proline, and methionine by photoemission spectroscopy. AB - The valence and core level photoelectron spectra of glycine, proline, and methionine in the gas phase have been investigated by VUV and soft X-ray radiation. The outer valence band photoemission spectra are similar to previously reported He I spectra, although relative peak intensities are different due to the different photon energy. We extended the spectral range to include the inner valence region. The carbon, nitrogen, and oxygen 1s as well as the sulfur 2p core level spectra of these amino acids have been measured and the states identified. Valence band spectra of proline have been recorded as a function of temperature, and they provide information about the relative populations of the lowest energy conformers. PMID- 17918920 TI - Contrasting singlet-triplet dynamical behavior of two vibrational levels of the acetylene S1 2(1)3(1)B2 polyad. AB - Surface electron ejection by laser-excited metastables (SEELEM) and LIF spectra of acetylene were simultaneously recorded in the regions of the A1Au-X1Sigmag+ nominal 2(1)3(1)4(2) Ka=1<--00 and 2(1)3(1)6(2) Ka=1<--00 bands near 46,140 cm( 1). The upper states of these two bands are separated by only approximately 100 cm(-1), and the two S1 vibrational levels are known to be strongly mixed by anharmonic and Coriolis interactions. Strikingly different patterns were observed in the SEELEM spectra in the regions of the 2(1)3(1)4(2) and 2(1)3(1)6(2) vibrational levels. Because the equilibrium structure of the T3 electronic state is known to be nonplanar, excitation of nu4 (torsion) and nu6 (antisymmetric in plane bend) are expected respectively to promote and suppress vibrational overlap between low-lying S1 and T3 vibrational levels. The nearly 50:50 mixed 2(1)3(1)4(2)-2(1)3(1)6(2) character of the S1 vibrational levels rules out this simple Franck-Condon explanation for the different appearance of the SEELEM spectra. A simple model is applied to the SEELEM/LIF spectra to explain the differences between spectral patterns in terms of a T3 doorway-mediated singlet triplet coupling model. PMID- 17918921 TI - Synthesis, potency, and in vivo profiles of quinoline containing histamine H3 receptor inverse agonists. AB - A new structural series of histamine H3 receptor antagonist was developed. The new compounds are based on a quinoline core, appended with a required basic aminoethyl moiety, and with potency- and property-modulating heterocyclic substituents. The analogs have nanomolar and subnanomolar potency for the rat and human H3R in various in vitro assays, including radioligand competition binding as well as functional tests of H3 receptor-mediated calcium mobilization and GTPgammaS binding. The compounds possessed favorable drug-like properties, such as good PK, CNS penetration, and moderate protein binding across species. Several compounds were found to be efficacious in animal behavioral models of cognition and attention. Further studies on the pharmaceutic properties of this series of quinolines discovered a potential problem with photochemical instability, an issue which contributed to the discontinuation of this series from further development. PMID- 17918922 TI - Synthesis of 1-deoxy-1-hydroxymethyl- and 1-deoxy-1-epi-hydroxymethyl castanospermine as new potential immunomodulating agents. AB - Two new C-1 epimeric hydroxymethyl castanospermine congeners 2a and 2b, synthesized by stereocontrolled intramolecular double reductive amination of D glucose derived beta-keto ester as a key step, showed impressive immuno potentiating property. The bioactivity was mediated through up-regulation of T(H1)/T(H2) cytokine ratio. The finding suggested that immunmodulatory activity of polyhydroxylated indolizidine alkaloids can be tuned by minor structural/stereochemical alterations. PMID- 17918923 TI - From docking false-positive to active anti-HIV agent. AB - Virtual screening of the Maybridge library of ca. 70 000 compounds was performed using a similarity filter, docking, and molecular mechanics-generalized Born/surface area postprocessing to seek potential non-nucleoside inhibitors of human immunodeficiency virus-1 (HIV-1) reverse transcriptase (NNRTIs). Although known NNRTIs were retrieved well, purchase and assaying of representative, top scoring compounds from the library failed to yield any active anti-HIV agents. However, the highest-ranked library compound, oxadiazole 1, was pursued as a potential "near-miss" with the BOMB program to seek constructive modifications. Subsequent synthesis and assaying of several polychloro-analogs did yield anti HIV agents with EC50 values as low as 310 nM. The study demonstrates that it is possible to learn from a formally unsuccessful virtual-screening exercise and, with the aid of computational analyses, to efficiently evolve a false positive into a true active. PMID- 17918924 TI - Tagged fragment method for evolutionary structure-based de novo lead generation and optimization. AB - Here we describe a computer-assisted de novo drug design method, EAISFD, which combines the de novo design engine EA-Inventor with a scoring function featuring the molecular docking program Surflex-Dock. This method employs tagged fragments, which are preserved substructures in EA-Inventor used for base fragment matching in Surflex-Dock, for constructing ligand structures under specific binding motifs. In addition, a target score mechanism is adopted that allows EAISFD to deliver a diverse set of desired structures. This method can be used to design novel ligand scaffolds (lead generation) or to optimize attachments on a fixed scaffold (lead optimization). EAISFD has successfully suggested many known inhibitor scaffolds as well as a number of new scaffold types when applied to p38 MAP kinase. PMID- 17918925 TI - Catalysis of zinc transfer by D-penicillamine to secondary chelators. AB - The antiarthritis drug D-penicillamine (D-PEN) catalyzes zinc(II) transfer from carboxypeptidase A to chelators such as thionein and EDTA at a rate constant up to 400-fold faster than the uncatalyzed release. Once D-PEN releases zinc(II) from enzyme stronger chelators can tightly bind zinc(II) leading to complete and essentially irreversible inhibition. D-PEN is the first drug to inhibit a zinc protease by catalyzing metal removal, and the name "catalytic chelation" is proposed for this mechanism. PMID- 17918926 TI - Designing compound subsets: comparison of random and rational approaches using statistical simulation. AB - Compound subsets, which may be screened where it is not feasible or desirable to screen all available compounds, may be designed using rational or random selection. Literature on the relative performance of random versus rational selection reports conflicting observations, possibly because some random subsets might be more representative than others and perform better than subsets designed by rational means, or vice versa. In order to address this likelihood, we simulated a large number of rationally designed subsets for evaluation against an equally large number of randomly generated subsets. We found that our rationally designed subsets give higher mean hit rates compared to those of the random ones. We also compared subsets comprising random plates with subsets of random compounds and found that, while the mean hit rate of both is the same, the former demonstrates more variation in the hit rate. The choice of compound file, rational subset method, and ratio of the subset size to the compound file size are key factors in the relative performance of random and rational selection, and statistical simulation is a viable way to identify the selection approach appropriate for a subset. PMID- 17918927 TI - Three data mining techniques to improve lazy structure-activity relationships for noncongeneric compounds. AB - We present three simple, yet effective data mining techniques for lazy structure activity relationships (SARs) of noncongeneric compounds. In lazy SARs, classifications are particularly tailored for each test compound. Therefore, it is possible to make the most of the structure of a test compound. In our case, we derive its substructures and use them to determine similar structures. To obtain a well-balanced and representative set of structural descriptors, we enrich this set by strongly activating or deactivating fragments from the training set and subsequently remove redundant fragments. Finally, we perform k-Nearest Neighbor classification for several values of k and take a vote among the resulting predictions. These techniques (enrichment, removing redundancy, and voting) are integrated into the system iSAR (instance-based structure-activity relationships) and tested individually to show the relative contribution to the system's performance. Experiments on three data sets indicate that this simple and lightweight approach performs at least on the same level as other, more complex approaches. PMID- 17918928 TI - Preparation, characterization, molecular and electronic structures, TDDFT, and TDDFT/PCM study of the solvatochromism in cyanovinylferrocenes. AB - Cis and trans isomers of ferrocene-based donor-acceptor assemblies, Fc C(I)=CH(I), Fc-C(I)=CH(CN), and Fc-C(CN)=CH(CN) (Fc is ferrocene), along with the Fc-C(CN)=C(CN)2 complex have been prepared and characterized by 1H, 13C, gHMQC, and gHMBC NMR spectra, IR, UV-vis, and MCD spectroscopy methods, as well as elemental analyses. The oxidation potentials, investigated by cyclic voltammetry, of all donor-acceptor assemblies are in agreement with the electron-acceptor strength of the substituents attached to the ferrocene core. X-ray crystallography studies of cis and trans isomers of Fc-C(I)=CH(CN) and Fc C(CN)=CH(CN) reveal a significant rotational flexibility of the cyanovinyl group, which was explained on the basis of semiempirical PM3 calculations. Electronic structures and solvatochromic properties of all complexes were investigated by UV vis spectroscopy, density functional theory (DFT), time-dependent DFT (TDDFT), and polarized continuum model (PCM) TDDFT approaches. The calculated vertical excitation energies and magnitudes of solvatochromic effect are consistent with the experimental data and clearly suggest the dominance of metal-to-ligand charge transfer bands in the visible region of the UV-vis spectra for all complexes. PMID- 17918929 TI - Synthesis and characterization of new nitrogen-donor-stabilized N silylphosphoranimine cations. AB - The phosphoranimine Br(CF3CH2O)2P=NSiMe3 (12) reacts quantitatively with nitrogen bases pyridine, 4,4'-bipyridine, and quinuclidine (quin) to form the N-donor stabilized phosphoranimine cations [N-donor.P(OCH2CF3)2=NSiMe3] ([15]+) in the presence of the halide abstractor AgOTf. In contrast to quinuclidine, in the absence of a halide abstractor, the weak bases pyridine and 4,4'-bipyridine do not undergo reactions with 12 or with the phosphoranimine Cl3P=NSiMe3 (7). Furthermore, unlike the weaker bases, quinuclidine also reacts with 7 to form the expected quinuclidine-stabilized phosphoranimine cation [quin.PCl2=NSiMe3]+ ([16]+) in the presence of AgOTf. However, in the absence of AgOTf, quinuclidine reacts with 7 to presumably yield the salt [16]Cl, which then undergoes a further quinuclidine ring-opening reaction to yield the cationic piperidyl-substituted phosphoranimine [(quin)CH2CH2C5H9N-PCl2=NSiMe3]Cl ([19]Cl). Reactions involving 7 and 12 with other halide abstraction reagents, such as GaCl3, are also described. PMID- 17918930 TI - Novel manganese(II) sulfonate-phosphonates with dinuclear, tetranuclear, and hexanuclear clusters. AB - Hydrothermal reactions of manganese(II) salts with m-sulfophenylphosphonic acid (m-HO3S-Ph-PO3H2, H3L) and 1,10-phenanthroline (phen) led to six novel manganese(II) sulfonate-phosphonates, namely, [Mn2(HL)2(phen)4][Mn2(HL)2(phen)4(H2O)](2).6H2O (1), [Mn4(L)2(phen)8(H2O)2][ClO4](2).3H2O (2), [Mn(phen)(H2O)4]2[Mn4(L)4(phen)4].10H2O (3), [Mn6(L)4(phen)8(H2O)2].4H2O (4), [Mn6(L)4(phen)8(H2O)2].24H2O (5), and [Mn6(L)4(phen)6(H2O)4].5H2O (6). The structure of 1 contains two types of dinuclear manganese(II) clusters, and 2-3 exhibit two types of tetranuclear manganese(II) cluster units. 4-5 feature two different types of isolated hexanuclear manganese(II) clusters, whereas the hexanuclear manganese(II) clusters in 6 are bridged by sulfonate-phosphonate ligands into a 1D chain. Magnetic property measurements indicate that there exist weak antiferromagnetic interactions between magnetic centers in all six compounds. PMID- 17918931 TI - DNA strand cleavage near a CC mismatch directed by a metalloinsertor. AB - Reagents for recognition and efficient cleavage of mismatched DNA without photoactivation were designed. They contain a combination of a mismatch-directing metalloinsertor, [Rh(bpy)2(chrysi)]3+ (bpy=2,2'-bipyridyl, chrysi=5,6 chrysenequinone diimine), and an oxidative cleavage functionality, [Cu(phen)2]+ (Cu). Both unconjugated (Rh+Cu) and conjugated (Rh-Cu) frameworks of the Rh insertor and Cu were prepared. Compared to Cu, both constructs Rh+Cu and Rh-Cu exhibit efficient site-specific DNA scission only with mismatched DNA, confirmed by experiments with 32P-labeled oligonucleotides. Furthermore, these studies indicate that DNA cleavage occurs near the mismatch in the minor groove and on both strands. Interestingly, the order of reactivity of the three systems with a CC mismatch is Rh+Cu>Rh-Cu>>Cu. Rh binding appears to direct Cu reactivity with or without tethering. These results illustrate advantages and disadvantages in bifunctional conjugation. PMID- 17918932 TI - Internuclear (31)P-(51)V distance measurements in polyoxoanionic solids using rotational echo adiabatic passage double resonance NMR spectroscopy. AB - We report the first results establishing rotational echo adiabatic passage double resonance (REAPDOR) experiments for distance measurements between a spin-1/2 (31P) and spin-7/2 (51V) pair in a series of vanadium-substituted polyoxoanionic solids from the Keggin and Wells-Dawson families. We have quantitatively measured 31P-51V distances in monovanadium substituted K4PVW11O40, 1-K7P2VW17O62, and 4 K7P2VW17O62. Numerical simulations of the experimental data yield very good agreement with the averaged P-W/P-V distances determined from the X-ray diffraction measurements in the same or related compounds. REAPDOR is therefore a very sensitive P-V distance probe anticipated to be especially useful in the absence of long-range order. Our results suggest that REAPDOR spectroscopy could be broadly applicable for interatomic distance measurements in other spin-7/2 spin-1/2 nuclear pairs. PMID- 17918934 TI - Palladium-catalyzed cross-coupling reactions in one-pot multicatalytic processes. AB - Palladium-catalyzed cross-coupling reactions have been investigated in multicatalytic processes to synthesize disubstituted alkenes and alkanes from carbonyl derivatives. The use of copper-catalyzed methylenation reactions is the key starting reaction to produce terminal alkenes which are not isolated, but submitted to further structure elongation. Not only is the isolation of the alkene intermediate unnecessary, but also the copper catalyst is a beneficial cocatalyst in the palladium-catalyzed cross-coupling reactions. The desired products are thus typically obtained in higher yields using this one-pot approach. We have used these processes to synthesize hydroxylated (E) stilbenoids, which are known chemopreventive and chemotherapeutic agents, odorant substituted indanes, and non-natural amino acids, such as homophenylalanine. PMID- 17918933 TI - Characterization of the maduropeptin biosynthetic gene cluster from Actinomadura madurae ATCC 39144 supporting a unifying paradigm for enediyne biosynthesis. AB - The biosynthetic gene cluster for the enediyne antitumor antibiotic maduropeptin (MDP) from Actinomadura madurae ATCC 39144 was cloned and sequenced. Cloning of the mdp gene cluster was confirmed by heterologous complementation of enediyne polyketide synthase (PKS) mutants from the C-1027 producer Streptomyces globisporus and the neocarzinostatin producer Streptomyces carzinostaticus using the MDP enediyne PKS and associated genes. Furthermore, MDP was produced, and its apoprotein was isolated and N-terminal sequenced; the encoding gene, mdpA, was found to reside within the cluster. The biosynthesis of MDP is highlighted by two iterative type I PKSs--the enediyne PKS and a 6-methylsalicylic acid PKS; generation of (S)-3-(2-chloro-3-hydroxy-4-methoxyphenyl)-3-hydroxypropionic acid derived from L-alpha-tyrosine; a unique type of enediyne apoprotein; and a convergent biosynthetic approach to the final MDP chromophore. The results demonstrate a platform for engineering new enediynes by combinatorial biosynthesis and establish a unified paradigm for the biosynthesis of enediyne polyketides. PMID- 17918936 TI - Spatial and intensity modulation of nanowire emission induced by mobile charges. AB - Single-molecule optical experiments carried out in conjunction with externally applied electric fields show deliberate spatial and intensity control over CdSe nanowire (NW) emission. In particular, by applying external fields to electrically isolated (single) NWs, their emission can be localized in areas of the wire closest to the positive electrode. In a few cases, the resulting emission intensity increases over the corresponding zero-field value by nearly an order of magnitude. More often than not, factors of 2-3 are seen. Reversing the field polarity causes the emission to localize in opposite regions of the wire. Emission from individual NWs can therefore be modulated. Complementary ac electric field measurements show that the effect persists up to 500 kHz. To explain the phenomenon, the effective passivation of surface trap states by mobile carriers is speculated. This, in turn, causes local changes in the NW emission quantum yield (QY). To verify the presence of such mobile charges, both ensemble and single NW bundle electrophoresis experiments are conducted. By investigating subsequent NW rotational and translational dynamics, an estimate for the number of mobile carriers is determined. A lower limit (best case) linear charge density of approximately 0.45-1.2 mobile electrons per micrometer of the wire is obtained. Apart from self-consistently explaining the field-induced NW emission modulation, the resulting data and subsequent analysis also suggests that the same mobile carriers may be the root cause of NW emission intermittency. Furthermore, given the ubiquity of stray charges, the resulting hypothesis may have additional applicability toward explaining blinking in other systems, a problem of current interest especially within the context of colloidal QDs. PMID- 17918935 TI - An artificial beta-sheet that dimerizes through parallel beta-sheet interactions. AB - This Article introduces a simple chemical model of a beta-sheet (artificial beta sheet) that dimerizes by parallel beta-sheet formation in chloroform solution. The artificial beta-sheet consists of two N-terminally linked peptide strands that are linked with succinic or fumaric acid and blocked along one edge with a hydrogen-bonding template composed of 5-aminoanisic acid hydrazide. The template is connected to one of the peptide strands by a turn unit composed of (S)-2 aminoadipic acid (Aaa). 1H NMR spectroscopic studies show that these artificial beta-sheets fold in CDCl3 solution to form well-defined beta-sheet structures that dimerize through parallel beta-sheet interactions. Most notably, all of these compounds show a rich network of NOEs associated with folding and dimerization. The compounds also exhibit chemical shifts and coupling constants consistent with the formation of folded dimeric beta-sheet structures. The aminoadipic acid unit shows patterns of NOEs and coupling constants consistent with a well-defined turn conformation. The present system represents a significant step toward modeling the type of parallel beta-sheet interactions that occur in protein aggregation. PMID- 17918937 TI - Synthetic ligands discovered by in vitro selection. AB - The recognition and catalytic properties of biopolymers derive from an elegant evolutionary mechanism, whereby the genetic material encoding molecules with superior functional attributes survives a selective pressure and is propagated to subsequent generations. This process is routinely mimicked in vitro to generate nucleic-acid or peptide ligands and catalysts. Recent advances in DNA-programmed organic synthesis have raised the possibility that evolutionary strategies could also be used for small-molecule discovery, but the idea remains unproven. Here, using DNA-programmed combinatorial chemistry, a collection of 100 million distinct compounds is synthesized and subjected to selection for binding to the N terminal SH3 domain of the proto-oncogene Crk. Over six generations, the molecular population converges to a small number of novel SH3 domain ligands. Remarkably, the hits bind with affinities similar to those of peptide SH3 ligands isolated from phage libraries of comparable complexity. The evolutionary approach has the potential to drastically simplify and accelerate small-molecule discovery. PMID- 17918938 TI - Resistive-pulse studies of proteins and protein/antibody complexes using a conical nanotube sensor. AB - There is increasing interest in using nanopores in synthetic membranes as resistive-pulse sensors for molecular and macromolecule analytes. In general, this method entails measuring current pulses associated with translocation of the analyte through the nanopore sensor element. A key challenge for this sensing paradigm is building selectivity into the protocol so that the current pulses for the target analyte can be distinguished from current pulses for other species that might be present in the sample. We show here that this can be accomplished with a protein analyte by adding to the solution an antibody that selectively binds the protein. We demonstrate this concept using bovine serum albumin (BSA) and a Fab fragment from a BSA-binding polyclonal antibody. Because the complex formed upon binding of the Fab to BSA is larger than the free BSA molecule, the current-pulse signature for the BSA/Fab complex can be easily distinguished from the free BSA. Furthermore, the BSA/Fab pulses can be easily distinguished from the pulses obtained for the free Fab and from pulses obtained for a control protein that does not bind to the Fab. Finally, we also show that the current pulse signature for the BSA/Fab complex can provide information about the size and stoichiometry of the complex. PMID- 17918939 TI - 1,4,5,8,9,12-Hexamethyltriphenylene. A molecule with a flipping twist. AB - The synthesis and characterization of 1,4,5,8,9,12-hexamethyltriphenylene (5) is described. In the solid state, X-ray crystallographic studies reveal that compound 5 presents a highly distorted C2 geometry with a 53 degrees end-to-end twist. In solution, variable-temperature 1H NMR studies and molecular modeling present a story of rapid dynamic conformational interconversions between two C2 enantiomers (with a low activation barrier) and a slower C2-D3 interconversion (with a relatively high barrier)--the first time clear evidence of conformational interchange for these hindered triphenylenes has been provided. Further studies have established that 5 is a fluorescent stable blue emitter, and that the compound undergoes an irreversible one-electron electrochemical oxidation. Calculations have predicted this to be a radical cation of C2 geometry with 60 degrees end-to-end twist. PMID- 17918940 TI - Activation of carbon-hydrogen bonds via 1,2-addition across M-X (X = OH or NH(2)) bonds of d(6) transition metals as a potential key step in hydrocarbon functionalization: a computational study. AB - Recent reports of 1,2-addition of C-H bonds across Ru-X (X = amido, hydroxo) bonds of TpRu(PMe3)X fragments {Tp = hydridotris(pyrazolyl)borate} suggest opportunities for the development of new catalytic cycles for hydrocarbon functionalization. In order to enhance understanding of these transformations, computational examinations of the efficacy of model d6 transition metal complexes of the form [(Tab)M(PH3)2X]q (Tab = tris-azo-borate; X = OH, NH2; q = -1 to +2; M = TcI, Re(I), Ru(II), Co(III), Ir(III), Ni(IV), Pt(IV)) for the activation of benzene C-H bonds, as well as the potential for their incorporation into catalytic functionalization cycles, are presented. For the benzene C-H activation reaction steps, kite-shaped transition states were located and found to have relatively little metal-hydrogen interaction. The C-H activation process is best described as a metal-mediated proton transfer in which the metal center and ligand X function as an activating electrophile and intramolecular base, respectively. While the metal plays a primary role in controlling the kinetics and thermodynamics of the reaction coordinate for C-H activation/functionalization, the ligand X also influences the energetics. On the basis of three thermodynamic criteria characterizing salient energetic aspects of the proposed catalytic cycle and the detailed computational studies reported herein, late transition metal complexes (e.g., Pt, Co, etc.) in the d6 electron configuration {especially the TabCo(PH3)2(OH)+ complex and related Co(III) systems} are predicted to be the most promising for further catalyst investigation. PMID- 17918941 TI - Conjugated polymer dots for multiphoton fluorescence imaging. PMID- 17918942 TI - Reversible formation and dispersion of chiral assemblies responding to electron transfer. PMID- 17918943 TI - Reticular synthesis of microporous and mesoporous 2D covalent organic frameworks. PMID- 17918945 TI - Synthesis of omega-hydroxy hexathiolate-protected subnanometric gold clusters. PMID- 17918944 TI - Stereospecificity of ketoreductase domains of the 6-deoxyerythronolide B synthase. AB - 6-Deoxyerythronolide B synthase (DEBS) is a modular polyketide synthase (PKS) responsible for the biosynthesis of 6-dEB (1), the parent aglycone of the broad spectrum macrolide antibiotic erythromycin. Individual DEBS modules, which contain the catalytic domains necessary for each step of polyketide chain elongation and chemical modification, can be deconstructed into constituent domains. To better understand the intrinsic stereospecificity of the ketoreductase (KR) domains, an in vitro reconstituted system has been developed involving combinations of ketosynthase (KS)-acyl transferase (AT) didomains with acyl-carrier protein (ACP) and KR domains from different DEBS modules. Incubations with (2S,3R)-2-methyl-3-hydroxypentanoic acid N-acetylcysteamine thioester (2) and methylmalonyl-CoA plus NADPH result in formation of a reduced, ACP-bound triketide that is converted to the corresponding triketide lactone 4 by either base- or enzyme-catalyzed hydrolysis/cyclization. A sensitive and robust GC-MS technique has been developed to assign the stereochemistry of the resulting triketide lactones, on the basis of direct comparison with synthetic standards of each of the four possible diasteromers 4a-4d. Using the [KS][AT] didomains from either DEBS module 3 or module 6 in combination with KR domains from modules 2 or 6 gave in all cases exclusively (2R,3S,4R,5R)-3,5-dihydroxy-2,4-dimethyl-n heptanoic acid-delta-lactone (4a). The same product was also generated by a chimeric module in which [KS3][AT3] was fused to [KR5][ACP5] and the DEBS thioesterase [TE] domain. Reductive quenching of the ACP-bound 2-methyl-3 ketoacyl triketide intermediate with sodium borohydride confirmed that in each case the triketide intermediate carried only an unepimerized d-2-methyl group. The results confirm the predicted stereospecificity of the individual KR domains, while revealing an unexpected configurational stability of the ACP-bound 2-methyl 3-ketoacyl thioester intermediate. The methodology should be applicable to the study of any combination of heterologous [KS][AT] and [KR] domains. PMID- 17918946 TI - Role of electrostatic interactions in transient encounter complexes in protein protein association investigated by paramagnetic relaxation enhancement. PMID- 17918947 TI - Enantioselective catalysis of ketoester-ene reaction of silyl enol ether to construct quaternary carbons by chiral dicationic palladium(II) complexes. PMID- 17918949 TI - Polysubstituted oxygen heterocycles by a Reformatsky-type reaction/reductive cyclization approach from enantiopure beta-ketosulfoxides. AB - The stereoselective synthesis of tetrasubstituted tetrahydrofurans and trisubstituted tetrahydropyrans bearing a sulfoxide was achieved by reductive cyclization (Et3SiH/TMSOTf) from the corresponding enantiopure hydroxy ketones protected as a dioxolane. These derivatives are easily accessible from a Reformatsky-type reaction between alpha-bromo-alpha'-sulfinyl ketones and protected alpha- or beta-ketoaldehydes, followed by diastereoselective reduction of the resulting beta-ketosulfoxide. PMID- 17918950 TI - Superior effect of a pi-acceptor ligand (phosphine-electron-deficient olefin ligand) in the Negishi coupling involving alkylzinc reagents. AB - Palladium-catalyzed Negishi cross-coupling involving primary and secondary alkyls, even in the presence of beta-H, can be achieved at ambient temperature using chelating ligands containing a phosphine and an electron-deficient olefin. The superior effects of the ligands were shown not only in the desired cross coupling product yields but also in the fast reaction at mild conditions. This reaction has been also scaled up to 50 g in 0.005 mol % catalyst (20,000 TONs) at room temperature. PMID- 17918951 TI - Pentacene oligomers and polymers: functionalization of pentacene to afford mono-, di-, tri-, and polymeric materials. AB - The synthesis and characterization of defined-length di- and trimeric pentacenes and the corresponding polymers are described. The synthesis is divergent from two common pentacene building blocks, 1 and 2, allowing for structural diversity. The resulting materials are air stable and exhibit good solubility in common organic solvents. PMID- 17918953 TI - A vector-selective reaction enables efficient construction of specific topology upon the primary side of beta-cyclodextrin. AB - N-dansylcysteines attached on the primary side of beta-cyclodextrin reacted with the saccharide hydroxyl groups in a vector-selective manner, affording the corresponding lactones. The desired topology of the lactones can be efficiently constructed simply by the selection of the proper enantiomer of D/L-cysteines. In comparison with the exo-lactone, the endo-lactone displayed 4 times stronger fluorescence intensity, stronger binding affinity to sodium adamantanecarboxylate, and 15 times larger signal changes in fluorescence intensity upon binding. PMID- 17918952 TI - Stereoselective total synthesis of bistramide A. AB - A highly stereoselective and convergent total synthesis of bistramide A is described. The salient feature of this synthesis is the construction of the spiroketal subunit by hydrolysis of dialkylated tosylmethyl isocyanide derivative derived via alkylation of TosMIC with suitably substituted halohydrin derivatives. PMID- 17918954 TI - Concise, stereoselective approach to the spirooxindole ring system of citrinadin A. AB - The spirooxindole ring system of citrinadin A has been synthesized with excellent control over the absolute stereochemistry at the spirocenter. The key step involves a novel diastereoselective DMDO-mediated oxidative rearrangement employing an 8-phenylmenthol chiral auxiliary on the indole nitrogen. PMID- 17918955 TI - Helical chirality transmission through a p-phenylene fragment in a hexa-lambda5 phosphazene. AB - The helical chirality of a conformationally stable macrobicyclic tri-lambda5 phosphazene is propagated through to its C3 symmetry axis and causes a measurable stereoinduction in the formation process of a second macrobicyclic tri-lambda5 phosphazene unit connected to the former one by a p-phenylene linker. PMID- 17918956 TI - Efficient preparation of N-phenylsulfenyl ketimines from oximes or nitro compounds without racemization of alpha-stereocenters. AB - As N-sulfenyl imines (e.g., RR'C=N-SAr) can be readily transformed to their N sulfinyl imines (RR'C=N-SOAr), N-sulfonyl imines (RR'C=N-SO2Ar), and N-sulfonyl oxaziridines, the very mild procedure developed to convert ketoximes and secondary nitro derivatives to N-arenesulfenyl ketimines constitutes a new and efficient route to all these series of compounds. The configuration of the alpha stereocenters is retained. PMID- 17918957 TI - Function of chlorophyll d in reaction centers of photosystems I and II of the oxygenic photosynthesis of Acaryochloris marina. AB - Reaction center chlorophylls (Chls) in photosystems II and I were studied in the isolated thylakoid membranes of a cyanobacterium, Acaryochloris marina, which contains Chls d and a as the major and minor pigments, respectively. The membranes contained PS I and II complexes at a 1.8:1 molar ratio on the basis of the spin densities on the tyrosine D radical and the photo-oxidized PS I primary donor (P740+). In the presence of ferricyanide, laser excitation induced bleach at 725 nm that recovered with time constants of 25 micros and 1.2 ms. The signal, designated P725, was suppressed by PS II inhibitors DCMU and hydroxylamine. The P725 spectrum was tentatively assigned to the absorption changes of the special pair Chl d, the accessory Chl d, and the acceptor pheophytin a in PS II. The addition of ascorbate induced the additional signal with a slow decay time constant of 4.5 ms. This signal showed a broad bleach at 740 nm and shift-type absorption changes at around 707 and 685 nm, which were assigned to the absorption changes of PS I special pair of Chl d (P740), the accessory Chl d, and the primary acceptor Chl a (A0), respectively. Mechanisms and the evolution of the Chl-d based reaction centers using far-red light are discussed together with the amino acid sequences of PS II D1 and D2 proteins. PMID- 17918958 TI - Role of glycosphingolipid conformational change in membrane pore forming activity of cobra cardiotoxin. AB - The major cardiotoxin from Taiwan cobra (CTX A3) is a pore forming beta-sheet polypeptide that requires sulfatide (sulfogalactosylceramide, SGC) on the plasma membrane of cardiomyocytes for CTX-induced membrane leakage and cell internalization. Herein, we demonstrate by fluorescence spectroscopic studies that sulfatides induce CTX A3 oligomerization in sulfatide containing phosphatidylcholine (PC) vesicles to form transient pores with pore size and lifetime in the range of about 30 A and 10(-2) s, respectively. These values are consistent with the CTX A3-induced conductance and mean lifetime determined previously by using patch-clamp electrophysiological experiments on the plasma membrane of H9C2 cells. We also derived the peripheral binding structural model of CTX A3-sulfatide complex in sulfatide containing PC micelles by NMR and molecular docking method and compared with other CTX A3-sulfatide complex structure determined previously by X-ray in membrane-like environment. The NMR results indicate that sulfatide head group conformation changes from a bent shovel (-sc/ap) to an extended (sc/ap) conformation upon initial binding of CTX A3. An additional global reorientation of sulfatide molecule is also needed for CTX A3 dimer formation as inferred by the difference between the X-ray and NMR complex structure. Since the overall folding of CTX A3 molecules remained the same, sulfatide in phospholipid bilayer is proposed to play an active role by involving its local and global conformational changes to promote both the oligomerization and reorientation of CTX A3 molecule for its transient pore formation and cell internalization. PMID- 17918959 TI - Ultrafast excited-state dynamics in the green fluorescent protein variant S65T/H148D. 1. Mutagenesis and structural studies. AB - Wild type green fluorescent protein (wt-GFP) and the variant S65T/H148D each exhibit two absorption bands, A and B, which are associated with the protonated and deprotonated chromophores, respectively. Excitation of either band leads to green emission. In wt-GFP, excitation of band A ( approximately 395 nm) leads to green emission with a rise time of 10-15 ps, due to excited-state proton transfer (ESPT) from the chromophore hydroxyl group to an acceptor. This process produces an anionic excited-state intermediate I* that subsequently emits a green photon. In the variant S65T/H148D, the A band absorbance maximum is red-shifted to approximately 415 nm, and as detailed in the accompanying papers, when the A band is excited, green fluorescence appears with a rise time shorter than the instrument time resolution ( approximately 170 fs). On the basis of the steady state spectroscopy and high-resolution crystal structures of several variants described herein, it is proposed that in S65T/H148D, the red shift of absorption band A and the ultrafast appearance of green fluorescence upon excitation of band A are due to a very short (10 kDa HAF fraction enabled three protein families, differing in average Mw, to be detected and may therefore be of potential value in the study of disorder-related variations in HAF protein profiles. The effects of freeze-drying, storage at -20 or -70 degrees C, and freeze-thawing cycles on HAF compositional stability were investigated, as well as stability at room temperature (to account for overnight data acquisition runs). These data are the basis for establishing statistically validated correlations between HAF NMR data and any physiological disorders of the fetus/mother. Freeze-drying caused signal loss for urea, ethanol, and compounds resonating at 2.22 and 1.17 ppm. Storage at -70 degrees C or lower is recommended since only minor compositional changes were observed, affecting mainly acetate and pyruvate. Freeze-thaw cycles did not cause significant compositional changes, and room-temperature stability studies indicated a 4-5 h maximum period of handling/acquisition time to ensure HAF stability. PMID- 17918969 TI - Carbohydrate analysis by desorption electrospray ionization fourier transform ion cyclotron resonance mass spectrometry. AB - We report the use of desorption electrospray ionization hybrid Fourier transform ion cyclotron resonance mass spectrometry (DESI-FT-ICR-MS) for the analysis of carbohydrates. Spectra of neat carbohydrates are presented along with their mass measurement accuracies and limits of detection. Furthermore, a comparison is made between the analyses of O-linked glycans from mucin by DESI-FT-ICR-MS and matrix assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry. Finally, glycans from mucin are identified by using the high mass measurement accuracy and tandem MS capabilities afforded by the hybrid FT ICR-MS platform. PMID- 17918971 TI - Self-assembly of monolayer-thick alumina particle-epoxy composite films. AB - Monolayer-thick composite films composed of alpha-alumina and Spurr's epoxy were prepared via a self-assembly process known as fluid forming. The process makes use of a high-spreading-tension fluid composed of volatile and nonvolatile components to propel particles across the air-water interface within a water bath. Continuous addition of the particle suspension builds a 2D particle film at the air-water interface. The spreading fluid compresses the film into a densely packed array against a submerged substrate. The assembled monolayer is deposited onto the substrate by removing the substrate from the bath. A dispersion containing a narrow size distribution, 10 microm alpha-alumina particles, light mineral oil, and 2-propanol was spread at the air-water interface and the alumina particles were assembled into densely packed arrays with an aerial packing fraction (APF) of 0.88. However, when mineral oil was replaced by Spurr's epoxy nonuniform films with low packing density resulted. It was found that replacing 2 propanol with a mixture of 2-propanol and 1-butanol with a volume ratio of 4:1 produced uniform, densely packed alumina/epoxy composite films. The role of the solvent mixture will be discussed. PMID- 17918972 TI - Effect of particle hydrophobicity on the properties of silica particle layers at the air-water interface. AB - This article describes a study of fumed silica particle layers adsorbed at the air-water interface. We have performed surface pressure, ellipsometry, and Brewster angle microscopy measurements. These determinations were complemented by surface viscoelasticity studies, using capillary waves to measure the compression moduli and an oscillating disc to measure the shear moduli. Our results show a strong influence of the particle hydrophobicity and surface density on the properties of the layers. Under compression-expansion, the particle layers rearrange quasi-instantaneously, and at high density, they buckle and/or collapse. Shear measurements show a transition from viscous to elastic behavior for particles with contact angles close to 90 degrees. The surface compression moduli are quite small and most likely not related to the stability of the foams made with these particles, in contrast to the case of more common surfactant foams. PMID- 17918970 TI - Fabrication, characterization, and application of boron-doped diamond microelectrodes for in vivo dopamine detection. AB - Highly boron-doped diamond (BDD) was deposited on chemically etched micrometer sized tungsten wires using microwave plasma assisted chemical vapor deposition (MPCVD), and these were used to fabricate BDD microelectrodes. BDD microelectrodes with very small diameter (about 5 microm) and 250 microm in length could be made successfully. In addition to the unique properties of BDD electrodes, such as a very low background current, high stability, and selective oxidation of dopamine (DA) in the presence of ascorbic acid (AA), other superior properties of the microelectrodes, including a constant current response, an increase in the mass transport, and the ability for use in high resistance media were also shown. An application study was conducted for in vivo detection of DA in mouse brain, where the BDD microelectrode was inserted into the corpus striatum of the mouse brain. A clear signal current response following medial forebrain bundle (MFB) stimulation could be obtained with high sensitivity. Excellent stability was achieved, indicating that the BDD microelectrodes are very promising for future in vivo electroanalysis. PMID- 17918973 TI - Alkyl chain length dependent hydrogelation of L-tryptophan-based amphiphile. AB - The search for low molecular weight hydrogelators (LMWHs) with varying structural motif is getting intense because of its potential application in biomedicines as well as the diversified area of nanobiotechnology. Hydrophobic interaction is one of the most crucial parameters in the design and development of such LMWHs. To this notion, a methodical investigation was carried out to find the influence of varying alkyl chain length of amphiphile on water gelation efficacy, which has been only marginally addressed in the literature to date. We have synthesized a series of low molecular weight L-tryptophan-based gelators, some of which are excellent gelator for plain water, an essential criterion for biological use. The alkyl chain induced hydrophobicity at the molecular level has remarkable influence in modulating water immobilization. Water gelation efficiency was improved more than 100 times on moving from 10 to 18 carbon atoms. The self aggregation behavior of these thermoreversible hydrogelators investigated through different spectroscopic and microscopic techniques showed that an optimum balance between hydrophilicity and hydrophobicity is indeed essential, which can be largely regulated by varying the alkyl chain length. Thus, the study offers better understanding toward tailoring the properties of gel in plain water and thereby paving the way for potential applications. PMID- 17918974 TI - Novel crown ether-capped cationic gold nanoclusters in an aqueous medium and their single-electron charging features. AB - On the basis of the formation of a noncovalent inclusion complex through crown ether-ammonium ion interaction, a new ligand, 18-crown-6-aminoethanethiol (18C6 AET), was predesigned and synthesized with a formation constant of 4.26 x 10(2) M(-1)s(-1) for the 1:1 noncovalent complex. Consequently, stable crown ether protected gold nanoclusters were synthesized with 18C6-AET in an aqueous medium with a rate of formation of 3.688 x 10(-4) min(-1). The newly formed 18C6-AET capped gold nanoclusters showed quantized double-layer charging, revealing their concentric capacitor structure, and effective cluster capacitance, C(CLU) was calculated to be 0.93 aF from the single-electron events. A larger C(CLU) in comparison to that of monolayer-protected gold clusters with longer chain lengths was attributed to a smaller ligand shell thickness as well as the cationic nature of the nanoclusters formed in an aqueous electrolyte medium. PMID- 17918975 TI - Regenerator dependent photoinduced desorption of a dicarboxylated cyanine dye from the surface of single-crystal rutile. AB - A photon-initiated desorption of a dicarboxylated thiacarbocyanine dye from a dye sensitized semiconducting oxide crystal has been observed when hydroquinone is used as a regenerator. No desorption was found under the same conditions when KI was used as the regenerator. Intermittent illumination experiments suggest that the oxidation products of the hydroquinone regenerator compete for dye adsorption sites. By comparing the photocurrent decay at both the dye monomer sensitization maximum and the dimer sensitization maximum, a rearrangement of monomer into dimer was observed. A kinetic model for the photocurrent decay as a function of desorption time was derived, and the desorption rate constants were obtained by fitting the experimental data to the model. PMID- 17918977 TI - Langmuir and Langmuir-Blodgett monolayers of molecular tweezers and clips. AB - Molecular clips and tweezers are able to selectively bind electron-deficient aromatic and aliphatic substrates. By means of pressure-area isotherms and Brewster angle microscopy (BAM), the self-association process and phase behavior of dimethylene-bridged molecular clips and tetramethylene-bridged molecular tweezers each substituted with two acetoxy groups as polar head groups were investigated. In a series of experiments, we observed that the molecular surface area of the clips and tweezers only depended on the skeletal structure and not on the polar groups. The measured areas agreed with the effective molecular diameters of the molecules if the aromatic side walls of the clips or tweezers were assumed to be aligned perpendicularly to the water surface. We compared the phase behavior of the pure molecular clips and tweezers with that of the host guest complexes of these molecules, which were formed with 1,2,4,5 tetracyanobenzene (TCNB) as the guest molecule. For the clips with a central benzene (I) and naphthalene spacer unit (II), the complex formation with TCNB had no measurable influence on the phase diagrams of the films. We observed, however, a dramatic difference in the BAM images and pi-A isotherms between the pure molecular tweezers III and its complex with TCNB (TCNB@III). In addition to the pi-A isotherms, we used the surface potential (V)-area (A) isotherms to compare the pure tweezers III with the corresponding complex (TCNB@III). There was a strong difference in the maximum surface potential value for the pure tweezers (450 mV) and that for the complex (300 mV). In additional experiments, we prepared LB layers of such molecules, which were investigated by fluorescence spectroscopy. In comparison to the pure tweezers III, a luminescence emission of charge-transfer (CT) origin was observed for the host-guest complex (TCNB@III) fixed on the solid substrate. It turned out that the spectra were in good agreement with the results observed in chloroform solution. PMID- 17918976 TI - Apparent dewetting of ultrathin multilayered polyelectrolyte films incubated in aqueous environments. AB - We have investigated and characterized changes in film morphology and surface structure that occur when ultrathin multilayered polyelectrolyte films fabricated from linear poly(ethylene imine) (LPEI), sodium poly(styrene sulfonate) (SPS), and two hydrolytically degradable polyamines (polymers 1 and 2) are incubated in physiologically relevant environments. Characterization of the physical erosion profiles of films having the structure (LPEI/SPS)10(1/SPS)4(2/SPS)4 (approximately 80 nm thick) by atomic force microscopy (AFM), reflective optical microscopy, and scanning electron microscopy (SEM) demonstrated that these materials undergo large-scale changes in surface structure and morphology upon incubation in phosphate-buffered saline (PBS) at 37 degrees C. The patterns and structures generated during this transformation (e.g., nucleation and growth of holes, coalescence of holes, formation of cell-type structures, and the subsequent breakup of these features into droplets) are similar in many ways to those observed for the dewetting of thin films of conventional polymers, such as polystyrene, on nonwetting surfaces. The processes reported here are sufficiently slow (they occur over approximately 100 h) and occur under sufficiently mild conditions (e.g., incubation in PBS at 37 degrees C) to permit characterization and quantification of the structures and features that arise during the course of these transformations. The apparent dewetting of these ultrathin films upon exposure to aqueous environments creates future opportunities to investigate and characterize processes of mass transport in this class of ionically cross-linked assemblies. PMID- 17918978 TI - Completely aqueous procedure for the growth of polymer brushes on polymeric substrates. AB - The growth of polymer brushes on polymer substrates is often challenging because of substrate incompatibility with the organic solvents used for initiator attachment. This letter reports the use of layer-by-layer adsorption of macroinitiators and subsequent aqueous ATRP from these immobilized initiators to prepare polymer brushes on polymeric substrates. Polyethersulfone (PES) films and porous membranes were modified with polyelectrolyte multilayer films, and a previously developed polycationic initiator, poly(2-(trimethylammonium iodide)ethyl methacrylate-co-2-(2-bromoisobutyryloxy)ethyl acrylate), was then electrostatically adsorbed onto these polyelectrolyte films. The immobilized macroinitiator is very efficient in initiating the growth of polymer brushes on PES, as demonstrated by aqueous syntheses of poly(2-hydroxyethyl methacrylate) (PHEMA) and poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) films. PHEMA (250 nm thick) and PDMAEMA (40 nm thick) brushes were grown in 2 h from surfaces modified with polycationic initiators. Moreover, this procedure is effective for growing brushes in the pores of PES membranes. PMID- 17918979 TI - Slippage of Newtonian liquids: influence on the dynamics of dewetting thin films. AB - Slippage of Newtonian liquids in the presence of a solid substrate is a newly found phenomenon, the origin of which is still under debate. In this article, we present a new analysis method to extract the slip length. Enhancing the slip of liquids is an important issue for microfluidic devices that demand for high throughput at low pumping power. We study the velocity of short-chained liquid polystyrene (PS) films dewetting from nonwettable solid substrates. We show how the dynamics of dewetting is influenced by slippage, and we compare the results of two types of substrates that give rise to different slip lengths. As substrates, Si wafers that have been coated with octadecyltrichlorosilane (OTS) or dodecyltrichlorosilane (DTS) were used. Our results demonstrate that the dewetting velocity for PS films on DTS is significantly larger than on OTS and that this difference originates from the different slip lengths of the liquid on top of the two surfaces. For PS films of thickness between 130 and 230 nm, we find slip lengths between 400 nm and 6 microm, depending on substrate and temperature. PMID- 17918980 TI - Adhesion of giant vesicles mediated by weak binding of sialyl-LewisX to E selectin in the presence of repelling poly(ethylene glycol) molecules. AB - Prior to establishing tight contact with the endothelium, cells such as leukocytes or cancer cells use the recognition between sialyl-LewisX ligands and E-selectin receptors to establish weak, reversible adhesion and to roll along the vessel wall. We study the physical aspects of this process by constructing a mimetic system that consists of a giant fluid vesicle with incorporated lipid anchored sialyl-LewisX molecules that bind to E-selectin that is immobilized on the flat substrate. The vesicles also carry a certain fraction of repelling PEG2000 molecules. We analyze the equilibrium state of adhesion in detail by means of reflection interference contrast microscopy and find that the adhesion process relies purely on the formation of one or more adhesion domains within the vesicle-substrate contact zone. We find that the content of ligands in the vesicle must be above 5 mol % to establish specific contacts. All concentrations of sialyl-LewisX above 8 mol % provide a very similar final state of adhesion. However, the size and shape of the adhesion domains strongly depend on both the concentrations of E-selectin (0-3500 molecules/microm2) and PEG2000 (0-5 mol %). At 3500 E-selectin molecules/microm2 and small concentrations of PEG2000, the vesicle-substrate contact is maximized and fully occupied by a single adhesion domain. At concentrations of 5 mol %, PEG2000 completely impedes the specific binding to any substrate. Lastly, an increase in the adhesion strength is observed in systems with identical compositions if the reduced volume of the vesicles is larger. PMID- 17918981 TI - Periodic mesoporous organosilicas with multiple bridging groups and spherical morphology. AB - Bi- and trifunctional periodic mesoporous organosilicas (PMOs) with phenylene, thiophene, and ethane bridging groups were synthesized using 1,2 bis(triethoxysilyl)ethane (BTEE), 1,4-bis(triethoxysilyl)benzene (BTEB), and 2,5 bis(triethoxysilyl)thiophene (BTET) organosilica precursors and a poly(ethylene oxide)-poly(D,L-lactic acid-co-glycolic acid)-poly(ethylene oxide) (PEO-PLGA-PEO) triblock copolymer template under low acidic conditions. The PMO samples with different concentrations of organic bridging groups were obtained in the form of spherical particles having average diameters of 2-3 mum and 2D hexagonal (p6m) mesostructure with pore diameters of 7.3-8.4 nm. The particle morphology and chemistry of pore walls were tailored using different mixtures of organosilica precursors. Adsorption and structural properties of the aforementioned PMOs have been studied by nitrogen adsorption and small-angle X-ray scattering, whereas their framework chemistry was quantitatively analyzed by solid-state 13C and 29Si MAS NMR. PMID- 17918982 TI - Polymer-induced synthesis of stable gold and silver nanoparticles and subsequent ligand exchange in water. AB - A simple, inexpensive, single-step synthesis of gold and silver nanoparticles using poly(allylamine) (PAAm) as a reducing and stabilizing agent is reported. The synthetic process was carried out in aqueous solution, making the method versatile and environmentally friendly. The synthesized polymer-stabilized nanoparticles are stable in water without particle aggregation at room temperature for at least a month. We demonstrate successful ligand exchange on the polymer-stabilized gold nanoparticles (AuNPs) with a variety of omega functionalized acid-, alcohol-, amine-, and biotin-terminated alkylthiols. The methodologies, including ligand exchange, also are applicable for the generation of finely dispersed silver nanoparticles. The synthesized gold and silver nanoparticles are characterized by UV-visible absorption spectroscopy and transmission electron microscopy (TEM). The different ligand-stabilized AuNPs are also analyzed by Fourier transform infrared (FTIR) spectroscopy. PMID- 17918983 TI - Conformational control of inorganic adhesion in a designer protein engineered for cuprous oxide binding. AB - Combinatorial selection of peptides that bind technological materials has emerged as a valuable tool for room-temperature nucleation and assembly of complex nanostructured materials. At present, the parameters that control peptide-solid binding are poorly understood, but such knowledge is needed to build the next generation of hybrid materials. Here, we use a derivative of the DNA binding protein TraI engineered with a disulfide-bonded cuprous oxide binding sequence called CN225 to probe the influence of sequence composition and conformation on Cu2O binding affinity. We previously reported a statistically significant enrichment in paired arginines (RR) among a family of cuprous oxide binding peptides and hypothesized that this is a key motif for binding. However, systematic alanine (A) substitutions in the CN225 RR motif (creating RA, AR, and AA pairs) do not support the hypothesis that RR is critical for Cu2O binding by CN225. Instead, we find that the presentation of the peptide in a disulfide constrained loop (i.e., the conformation present during combinatorial selection) is crucial for binding to the metal oxide. Our results suggest that caution should be exerted when extrapolating from statistical data and that, in some cases, conformation is more important than composition in determining peptide inorganic adhesion. PMID- 17918984 TI - Proteomic analysis reveals differences in protein expression in spheroid versus monolayer cultures of low-passage colon carcinoma cells. AB - Spheroid cultures of cancer cells may better reflect characteristics of tumors than traditional monolayer cultures. Furthermore, low-passage cancer cell lines recapitulate the properties of the original tumor cells more closely than commonly used standard cell lines that experience artificial selection processes and mutations over years of passaging. Here we established spheroid cultures of the low-passage colon cancer cell line COGA-5 and stable COGA-12 aggregates with local areas of compaction. The proteomes of both three-dimensional cultures were analyzed versus their corresponding two-dimensional cultures. 2-D gel electrophoresis followed by peptide mass fingerprinting identified three differently expressed proteins in COGA-5 spheroids (acidic calponin, hydroxyprostaglandin dehydrogenase, and lamin A/C) and two in COGA-12 partly compact aggregates (two isoelectric variants of the acidic ribosomal protein P0) compared to the respective monolayer cultures. The lamin A/C spot showed a lower molecular weight in the 2-D gel (30 kDa) than expected for full-length lamin. Further Western blot analysis and immunocytochemistry identified the lamin protein as a caspase-6-cleavage product in apoptotic cells of the spheroid. Similar caspase-dependent lamin cleavage was observed in monolayer cultures after serum withdrawal and further increased under hypoxic conditions, suggesting cleaved lamin as an indicator for apoptotic stress. In conclusion, proteome analysis of multicellular spheroids versus monolayers cultures identifies differential protein expression relevant to tumor cell proliferation, survival, and chemoresistance and thus may reveal novel targets for cancer therapy. PMID- 17918985 TI - Exploring the platelet proteome via combinatorial, hexapeptide ligand libraries. AB - A combinatorial ligand library, composed of millions of diverse hexapeptide baits, able to capture and concentrate the "low-abundance" proteome while drastically cutting the concentration of the most abundant species, has been applied to the exploration of the soluble platelet proteome. Mass spectrometry analysis of untreated and library-treated platelets has resulted in the identification of 435 unique gene products. Of those, 147 entries (35% of the total) have not been described among the list of >1100 proteins in proteomic platelet investigations reported before. In addition, the analysis of excised spots from two-dimensional electrophoresis analysis allowed 57 other proteins to be added that were not found in LC-MS analysis, 33 of them not described before in proteomics studies, bringing the total number of new gene products to 180. Thus, the present data add a non-negligible number of species for continuing the "cartography" of the proteomic asset of platelets, in view of completing the mapping procedure for a deeper understanding of the physiology and pathology of this blood cell. Because the capturing process is performed under physiological conditions, by exploiting, for binding to the combinatorial library, the native protein configuration, the described technique is not adapted to capture highly hydrophobic proteins, which need strong denaturing and solubilizing conditions that are incompatible with our working procedure. Thus, our list reports essentially hydrophilic proteins, with negative GRAVY indexes. PMID- 17918986 TI - Organellar proteomics of human platelet dense granules reveals that 14-3-3zeta is a granule protein related to atherosclerosis. AB - Dense granules, a type of platelet secretory organelle, are known to accumulate high concentrations of small molecules such as calcium, adenine nucleotides, serotonin, pyrophosphate, and polyphosphate. Protein composition of these granules has been obscure, however. In this paper, we use proteomics techniques to describe, for the first time, the soluble protein composition of platelet dense granules. We have isolated highly enriched human platelet dense granule fractions that have been analyzed using two proteomics methods. Using this approach, we have identified 40 proteins, and most of them, such as actin associated proteins, glycolytic enzymes, and regulatory proteins, have not previously been related to the organelle. We have focused our efforts on studying 14-3-3zeta, a member of a conserved family of proteins that interact with hundreds of different proteins. We have demonstrated that 14-3-3zeta is localized mostly on dense granules and that it is secreted after platelet activation. As some proteins secreted from activated platelets could promote the development of atherosclerosis and thrombosis, we have studied the expression of 14-3-3zeta in sections of human abdominal aorta of patients with aneurysm, identifying it at the atherosclerotic plaques. Together, our results reveal new details of the composition of the platelet dense granule and suggest an extracellular function for 14-3-3zeta associated with atherosclerosis. PMID- 17918987 TI - Effect of surfactant on water-soluble conjugated polymer used in biosensor. AB - The effect of nonionic surfactants on the cationic conjugated polymer (CCP), poly{9,9-bis[6-(N,N-trimethylammonium)hexyl]fluorene-co 1,4-phenylene} iodide 1, has been investigated. It is shown that the CCP in various solvents exists in three phases: isolated polymer chains, polymer aggregate, and variable size clusters (partially dissolved polymer). It is shown that nonionic surfactants enhance the photoluminescence (PL) quantum yield of the CCP in water by breakup of polymer aggregates, which eliminates the nonemissive interchain quenching with aggregates and increases surface-to-volume ratio of the CCP. Furthermore, the surfactants reduce quenching by incorporation of the CCP into aggregates or binary micelles. Surfactant also reduces the polar interaction strength between CCP and water and enhances CCP quenching by the counterions (iodine) by ion pairing effect. The dynamics of the interactions are complex and reveal that the surfactant induces rapid increase in the PL which imply that the main force that causes the aggregation is weak and may be due to hydrophobic interaction of the CCP in water rather than a solid, particulate-like state. Time-resolved fluorescence measurements at the exciton energy (420 nm) confirm that the CCP in water and in some organic solvents is a multiphase system in which three exponential decay terms are needed to fit the decay profile of the CCP. The change in the decay lifetime explains clearly the effect of surfactant and solvent polarity on the three CCP phases. The average lifetime of the CCP does not increase with surfactant, but the number of isolated polymer chains increases which leads to higher PL quantum yield. The association between the polymer and a quencher, single-strand deoxyribonucleic acid (ssDNA), was investigated. It indicated that CCP:ssDNA forms a weak electrostatic complex that does not alter the absorption spectra of the CCP but induces a strong CCP fluorescence quenching with association constant KS = 5 x 10(7) M(-1). At low ssDNA concentrations, the surfactant reduces quenching in the complex possibly by preventing charge transfer processes. This may be due to an increase in the distance between the CCP and ssDNA through incorporation of the CCP into aggregates (micelles). However, at high ssDNA concentration, the quenching increases sharply which may be assigned to the increase in the electrostatic force destroying the micelles' structure around the CCP, leading to contact quenching as well as DNA induced CCP aggregation, which in turn leads to CCP-CCP quenching. PMID- 17918988 TI - Small angle neutron scattering study of polyelectrolyte conformation incorporated into hybrid threadlike micelles under strong shear flows. AB - A small angle neutron scattering study revealed that polyelectrolytes, sodium salts of partially sulfonated polystyrenes with narrow distributed molar masses of Mn = 27 x 10(3) and 340 x 10(3), which were incorporated into hybrid threadlike micelles formed with cetyltrimethylammonium bromide in aqueous (deuterium oxide) solutions at a quiescent state, behaved as rigid rods with lengths of 16 and 200 nm and the same radius of 2.3 approximately 2.4 nm, respectively. Under strong shear flows at shear rates much higher than the reciprocal of the mechanical relaxation time for the solution, the formed hybrid threadlike micelles were highly orientated to the shear flow axis owing to the generation of a shear induced liquid crystalline phase. The polyelectrolytes incorporated into the highly orientated micelle also maintained essentially the same conformation as those in the randomly orientated micelles in a quiescent state even at high shear rates. PMID- 17918989 TI - On the atomistic mechanisms of alkane (methane-pentane) separation by distillation: a molecular dynamics study. AB - Insights into the liquid-vapor transformation of methane-pentane mixtures were obtained from transition path sampling molecular dynamics simulations. This case study of the boiling of non-azeotropic mixtures demonstrates an unprejudiced identification of the atomistic mechanisms of phase separation in the course of vaporization which form the basis of distillation processes. From our simulations we observe spontaneous segregation events in the liquid mixture to trigger vapor nucleation. The formation of vapor domains stabilizes and further promotes the separation process by preferential evaporation of methane molecules. While this discrimination holds for all mixtures of different composition studied, a full account of the boiling process requires a more complex picture. At low methane concentration the nucleation of the vapor domains includes both methane and pentane molecules. The pentane molecules, however, tend to form small aggregates and undergo rapid re-condensation within picoseconds to nanoseconds scales. Accordingly, two aspects of selectivity accounting for methane-pentane separation in the course of liquid-vapor transformations were made accessible to molecular dynamics simulations: spontaneous segregation in the liquid phase leading to selective vapor nucleation and growth favoring methane vaporization and selective re-condensation of pentane molecules. PMID- 17918990 TI - New short aliphatic chain ionic liquids: synthesis, physical properties, and catalytic activity in aldol condensations. AB - This paper reports on the synthesis of new short aliphatic chain ionic liquids and the study of the temperature dependence of density, ultrasonic velocities, and ionic conductivity in the range of 278.15-338.15 K. Fourier transform infrared spectra establishes their simple ionic salt structure. Because of their polarity, the ionic liquids are able to dissolve polar solvents and inorganic salts, all of them showing high tolerance in hydroxylic media. The observed temperature trend of the studied properties points out the special packing of these ionic liquids, as well as the strong influence of the steric hindrance among linear aliphatic residues enclosed in anions and cations. One of them showed a very high melting temperature. A collection of slightly basic ionic liquids were used to test their catalytic activity in several aldol condensation reactions of some carbonyl compounds. The best conversions and selectivities were obtained using single ionic liquids, with no synergetic effects being observed when different concentrations of mixed ionic liquids were used as catalysts. In any case, the ionic liquid can also easily be recycled from reaction media, suggesting a promising method of process design for this kind of reaction. PMID- 17918991 TI - Vibrational analysis of amino acids and short peptides in hydrated media. II. Role of KLLL repeats to induce helical conformations in minimalist LK-peptides. AB - Aqueous solution secondary structures of minimalist LK-peptides, with the generic sequence defined as KLL(KLLL)nKLLK, have been analyzed by means of circular dichroism (CD) and Raman scattering techniques. Our discussion in the present paper is mainly focused on four synthetic peptides (from 5 to 19 amino acids), KLLLK, KLLKLLLKLLK, KLLKLLLKLLLKLLK, and KLLKLLLKLLLKLLLKLLK, corresponding to the repeat unit, and to the peptide chains with the values of n = 1-3, respectively. CD and Raman spectra were analyzed in order to study both structural features of the peptide chains and their capability to form aggregates. On the basis of the obtained results it was concluded that the conformational flexibility of the shortest peptides (5-mer and 11-mer) is high enough to adopt random, beta-type, and helical chains in aqueous solution. However, the 11-mer shows a clear tendency to form beta-strands in phosphate buffer. The conformational equilibrium can be completely shifted to beta-type structures upon increasing ionic strength, i.e., in PBS and tris buffers. This equilibrium can also be shifted toward helical chains in the presence of methanol. Finally, the longest peptides (15-mer and 19-mer) are shown to form alpha-helical chains with an amphipathic character in aqueous solution. The possibility of bundle formation between helical chains is discussed over the temperature-dependent H-D exchange on labile hydrogens and particularly by considering the particular behavior of an intense Raman mode at 1127 cm-1 originating from the leucine residue side chain. The conformational dependence of this mode observed upon selective deuteration has never been documented up to now. PMID- 17918994 TI - Optical and electrochemical detection of saccharides with poly(aniline-co-3 aminobenzeneboronic acid) prepared from enzymatic polymerization. AB - Boronic acid-based sensors for saccharides have been developed via biocatalysis. The self-doped copolymer of poly(aniline-co-3-aminobenzeneboronic acid) [poly(aniline-co-AB)], with various mole ratios of two components, was synthesized by oxidative enzymatic polymerization using a natural biocatalyst such as horseradish peroxidase together with an anionic polyelectrolyte template (sulfonated polystyrene) under mild conditions (pH 4.5). Poly(aniline-co-AB), having an aniline boronic acid-to-aniline ratio of 1:2 on average, gave rise to a green doped polymer with absorption maxima at 745 nm. The potentiometric detection of saccharides using poly(aniline-co-AB) is presented. Characteristics of both transient and steady-state response associated with the complex formation of poly(aniline-co-AB) with various saccharides were monitored by UV-vis spectroscopy and cyclic voltammetry (CV). The results obtained from UV-vis spectroscopy and CV show that the sensitivity of enzymatically synthesized water soluble poly(aniline-co-AB) for various saccharides was improved significantly compared to the chemically synthesized counterpart. A possible mechanism for the sensitive detection of sugar molecules by boronic acid is proposed on the basis of UV-vis and IR spectrophotometry, and four-point probe conductivity measurements. PMID- 17918992 TI - Vibrational spectra of C60.C8H8 and C70.C8H8 in the rotor-stator and polymer phases. AB - C(60).C(8)H(8) and C(70).C(8)H(8) are prototypes of rotor-stator cocrystals. We present infrared and Raman spectra of these materials and show how the rotor stator nature is reflected in their vibrational properties. We measured the vibrational spectra of the polymer phases poly(C(60)C(8)H(8)) and poly(C(70)C(8)H(8)) resulting from a solid-state reaction occurring on heating. On the basis of the spectra, we propose a connection pattern for the fullerene in poly(C(60)C(8)H(8)), where the symmetry of the C(60) molecule is D(2h). On illuminating the C(60).C(8)H(8) cocrystal with green or blue light, a photochemical reaction was observed leading to a product similar to that of the thermal polymerization. PMID- 17918995 TI - Characterization of tissue response and in vivo degradation of cholecyst-derived extracellular matrix. AB - A rat subcutaneous implantation model was used to evaluate the in vivo degradation and tissue response of cholecyst-derived extracellular matrix (CEM). This response was compared to that of glutaraldehyde (GA) cross-linked CEM and porcine heart valve (HV), which are designated as GAxCEM and GAxHV, respectively. Tissue composition, inflammatory cell distribution, and angiogenesis at the implant site were quantified using stereological parameters, thickness (Ta), volume fraction (Vv), surface density (Sv), length density (Lv), and radius of diffusion (Rdiff). CEM was completely infiltrated with host tissue at 21 days and resorbed by 63 days. GAxCEM was also infiltrated with host tissue, while GAxHV matrix was impermeable to host tissue infiltration. Both GAxCEM and GAxHV retained their scaffold integrity until 63 days with no apparent degradation. A fibrous tissue of thickness <52 mum, rich in collagen and vasculature, surrounded all scaffolds, and from 21 to 63 days the fibrous tissue showed maturation with a significant increase in their fibrocyte content. No signs of acute inflammatory response were observed in the study period for any of the scaffolds, while the chronic inflammatory response was predominated with macrophages for all scaffolds except for CEM at 63 days. A higher degree of giant cell formation was observed with GA cross-linked scaffolds. From 21 to 63 days, lymphocytic response decreased for CEM, while it increased significantly for GAxHV. Angiogenesis/neo vascularization was uniform for CEM (reaching the core), significantly lower for GAxCEM within the implant area as compared to CEM, while restricted to the exterior of GAxHV matrix. In summary, CEM was a fast degrading scaffold that induced a transitional inflammatory response accompanied by gradual resorption and replacement by host connective tissue as compared to the very slow degrading GA cross-linked controls, GAxCEM and GAxHV, which caused a sustained chronic inflammatory response and remained at the site of implantation until the end of the study period of 63 days. PMID- 17918997 TI - Kinetic evidence for the formation of monocationic N,N'-disubstituted phthalamide in tertiary amine-catalyzed hydrolysis of N-substituted phthalimides. AB - A kinetic study on the aqueous cleavage of N-(2-methoxyphenyl)phthalimide (1) and N-(2-hydroxyphenyl)phthalimide (2), under the buffers of N-methylmorpholine, reveals the equilibrium presence of monocationic amide (Ctam) formed due to nucleophilic reactions of N-methylmorpholine with 1 and 2. Pseudo-first-order rate constants for the reactions of water and HO- with Ctam (formed through nucleophilic reaction of N-methylmorpholine with 1) are 4.60 x 10(-5) s-1 and 47.9 M-1 s-1, respectively. But the cleavage of Ctam, formed through nucleophilic reaction of N-methylmorpholine with 2, involves intramolecular general base (2'-O group of Ctam)-assisted water attack at carbonyl carbon of cationic amide group of Ctam in or before the rate-determining step. PMID- 17918996 TI - Deoxygenative [1,2]-hydride shift rearrangements in nucleoside and sugar chemistry: analogy with the [1,2]-electron shift in the deoxygenation of ribonucleotides by ribonucleotide reductases. AB - A variant of the semipinacol rearrangement that was observed in our laboratory has been applied to the synthesis of several furanose and pyranose derivatives. The process consists of an "orchestrated" [1,2]-hydride shift with departure of a leaving group from the opposite face. Transient formation of a C=O group is followed by rapid transfer of a hydride-equivalent from the same face from which the leaving group departed, which results in double inversion of stereochemistry at the two vicinal carbon atoms. Treatment of 2'-O- and 3'-O-tosyladenosine with lithium triethylborohydride in DMSO/THF gave the respective 2'- and 3' deoxynucleoside analogues with beta-D-threo configurations. Identical treatment of 5'-O-TPS-2'-O-tosyladenosine gave 9-(5-O-TPS-2-deoxy-beta-D-threo pentofuranosyl)adenine. The same [1,2]-hydride shift and stereochemistry with the 5'-OH and 5'-O-TPS compounds demonstrated the absence of remote hydroxyl-group participation. Application of this process to other nucleoside 2'-O-tosyl derivatives gave the 2'-deoxy-threo compounds in good yields. The reaction-rate order was OTs approximately Br >> Cl for 2'-O-tosyladenosine, 2'-bromo-2' deoxyadenosine, and 2'-chloro-2'-deoxyadenosine (all with beta-d-ribo configurations). Analogous results were obtained with mannopyranoside derivatives with either 4,6-O-benzylidene protection or a free OH group at C4. Deuterium labeling clearly defined the stereochemical course as a cis-vicinal [1,2]-hydride shift on the face opposite to the original cis OH and OTs groups followed by hydride transfer from the face opposite to the [1,2]-hydride shift. Synthetic and mechanistic considerations are discussed. PMID- 17918998 TI - Dihalocarbene insertion reactions into C-H bonds of compounds containing small rings: mechanisms and regio- and stereoselectivities. AB - Novel insertion reactions of dichloro- and dibromocarbene into carbon-hydrogen bonds adjacent to cyclopropane rings are reported. It is found that the predominant isomers formed in the reactions with bicyclo[4.1.0]heptane result from insertion into the endo carbon-hydrogen bonds alpha to the three-membered ring. In the reactions of bicyclo[3.1.0]hexane, however, the exo dihalocarbene insertion products are formed as the major isomers. In some compounds cyclopropane rings "activate" adjacent carbon-hydrogen bonds, whereas other systems containing three-membered rings do not. Moreover, the influence of various substituents (methyl, geminal dimethyl, phenyl, methoxy, and ethoxy) attached to bicyclo[3.1.0]hexane and bicyclo[4.1.0]heptane in dihalocarbene reactions has been studied. The findings can be explained by the concept of maximum orbital overlaps of Walsh orbitals of the cyclopropane rings and the alpha carbon-hydrogen bonds. In stark contrast, selective insertion into the tertiary carbon-hydrogen bonds of the cyclobutane ring in bicyclo[4.2.0]octane is observed. PMID- 17918999 TI - Solvent-controlled stereoselective formation of a cyclic ether in the lewis acid mediated allylation of an alpha-chloroacetoxy acyclic ether. Very high stereoselectivity in CH3CN vs low stereoselectivity in CH2Cl2. AB - The MgBr2.OEt2-mediated intramolecular allylation of a 4:1 diastereoisomeric mixture of the alpha-chloroacetoxyl ether 1a bearing the A-G/JK ring system of brevetoxin B in CH2Cl2 gave a 1:1 diastereoisomeric mixture of the trans- and cis cyclization products 4a and 5a having the A-G/I-K ring system, while that in CH3CN afforded the trans-isomer 4a nearly as the single product. To help clarify a reason for this marked solvent effect in the cyclization of the brevetoxin B precursor, DFT computations for the starting materials, intermediates, transition states, and products were carried out. The cyclization would proceed through a carbocation intermediate 3a having sp2 flat structure (SN1 type mechanism) in CH2Cl2, in which the activation energies leading to both diastereoisomers are approximately identical, while in CH3CN alkylnitrilium salts 6a would be formed through the coordination of CH3CN to the carbocation leading to an sp3-type intermediate in which sever steric hindrance takes place in the transition state leading to the undesired diastereoisomer. The scope of this novel solvent controlled stereoselectivity was tested for simple compounds. In small model compounds the marked solvent dependence was absent, but the model bearing two consecutive cyclic ether rings 1b exhibited a remarkable solvent effect similar to that observed in the brevetoxin B system. PMID- 17919000 TI - Organocatalytic asymmetric robinson annulation of alpha,beta-unsaturated aldehydes: applications to the total synthesis of (+)-palitantin. AB - The highly enantioselective organocatalytic Robinson annulation of alpha,beta unsaturated aldehydes was achieved, catalyzed by l-proline and trialkylamines and providing the formal [4 + 2] cycloaddition adducts. Additionally, in some examples in the catalysis with diarylpyrrolinol silyl ethers, the reactions afforded the [4 + 2] adducts with high enantioselectivity (>99.5% ee). The structure of the adduct, obtained from the reaction of 3-methylbut-2-enal and (E) 3-(2-nitrophenyl)acrylaldehyde, was confirmed by X-ray analysis. The absolute configurations of some [4 + 2] cycloadducts were investigated, and the methodology was applied in the synthesis of (+)-palitantin. PMID- 17919001 TI - Highly enantioselective allylation of alpha-ketoesters catalyzed by N,N'-dioxide In(III) complexes. AB - An efficient asymmetric allylation of alpha-ketoesters was catalyzed by the N,N' dioxide-In(III) complex in excellent yields (up to 99%) and high enantioselectivities (up to 94% ee) for a variety of substrates under mild reaction conditions. On the basis of experimental results, a possible catalytic cycle including a transition state has been proposed to explain the origin of the reactivity and asymmetric inductivity, and a bifunctional catalysis was described with Lewis base N-oxide activating tetraallylstannane and Lewis acid indium activating alpha-ketoester. PMID- 17919002 TI - The cycloaddition of nitrones with homochiral cyclopropanes. AB - The cycloaddition of nitrones and enantiomerically pure cyclopropane-1,1 dicarboxylates is examined. Transfer of optical activity to the adduct is dictated by thermal reaction conditions and nature of cyclopropane substitution. Optically active 2-substituted cyclopropane-1,1-dicarboxylates are shown to racemize under typical reaction conditions, providing evidence for a zwitterionic ring-opened intermediate. PMID- 17919003 TI - Synthesis of functionalized 1,2-dihydroisoquinolines via multicomponent one-pot reaction of 2-alkynylbenzaldehyde, amine, zinc, and allylic bromide or benzyl bromide. AB - A multicomponent one-pot reaction of 2-alkynylbenzaldehydes, amines, zinc, and allylic bromide or benzyl bromide using the combination of Mg(ClO4)2/Cu(OTf)2 as catalyst in THF/DCE (1:20) is described, which provides an efficient and practical route for the synthesis of functionalized 1,2-dihydroisoquinolines. PMID- 17919004 TI - Molecular fragmentation driven by ultrafast dynamic ionic resonances. AB - The authors time resolve molecular motion in bound state, ionic potentials that leads to bond cleavage during the interaction with intense, ultrafast laser fields. Resonances in molecular ions play an important role in dissociative ionization with ultrafast laser fields, and the authors demonstrate how these resonances evolve in time to produce dissociation after initial strong-field ionization. Exploiting such dynamic resonances offers the possibility of controlled bond breaking and characterizing time-dependent molecular structure. PMID- 17919005 TI - Resonance vibrational Raman optical activity: a time-dependent density functional theory approach. AB - We present a method to calculate both on- and off-resonance vibrational Raman optical activities (VROAs) of molecules using time-dependent density functional theory. This is an extension of a method to calculate the normal VROA by including a finite lifetime of the electronic excited states in all calculated properties. The method is based on a short-time approximation to Raman scattering and is, in the off-resonance case, identical to the standard theory of Placzek. The normal and resonance VROA spectra are calculated from geometric derivatives of the different generalized polarizabilites obtained using linear response theory which includes a damping term to account for the finite lifetime. Gauge origin independent results for normal VROA have been ensured using either the modified-velocity gauge or gauge-included atomic orbitals. For the resonance VROA only the modified-velocity gauge has been implemented. We present some initial results for H(2)O(2) and (S)-methyloxirane and compare with predictions from a simple two-state approximation. PMID- 17919006 TI - Comparison of double-ended transition state search methods. AB - While a variety of double-ended transition state search methods have been developed, their relative performance in characterizing complex multistep pathways between structurally disparate molecular conformations remains unclear. Three such methods (doubly-nudged elastic band, a string method, and a growing string method) are compared for a series of benchmarks ranging from permutational isomerizations of the seven-atom Lennard-Jones cluster (LJ(7)) to highly cooperative LJ(38) and LJ(75) rearrangements, and the folding pathways of two peptides. A database of short paths between LJ(13) local minima is used to explore the effects of parameters and suggest reasonable default values. Each double-ended method was employed within the framework of a missing connection network flow algorithm to construct more complicated multistep pathways. We find that in our implementation none of the three methods definitively outperforms the others, and that their relative effectiveness is strongly system and parameter dependent. PMID- 17919007 TI - Time-dependent density functional theory calculation of van der Waals coefficient of sodium clusters. AB - In this paper we employ all-electron ab initio time-dependent density functional theory based method to calculate the long range dipole-dipole dispersion coefficient (van der Waals coefficient) C(6) of sodium atom clusters containing even number of atoms ranging from 2 to 20 atoms. The dispersion coefficients are obtained via Casimir-Polder relation [Phys. Rev. 3, 360 (1948)]. The calculations are carried out with two different exchange-correlation potentials: (i) the asymptotically correct statistical average of orbital potential (SAOP) and (ii) Vosko-Wilk-Nusair representation [Can. J. Phys. 58, 1200 (1980)] of exchange correlation potential within local density approximation. A comparison with the other theoretical results has been performed. We also present the results for the static polarizabilities of sodium clusters and also compare them with other theoretical and experimental results. These comparisons reveal that the SAOP results for C(6) and static polarizability are quite accurate and very close to the experimental results. We examine the relationship between volume of the cluster and van der Waals coefficient, and find that to a very high degree of correlation C(6) scales as the square of the volume. We also present the results for van der Waals coefficient corresponding to cluster-Ar atom and cluster-N(2) molecule interactions. PMID- 17919008 TI - Local electrostatics algorithm for classical molecular dynamics simulations. AB - An implementation of a local, linear scaling algorithm for computing electrostatic interactions in molecular dynamics simulations that was recently proposed [J. Rottler and A. C. Maggs, Phys. Rev. Lett. 93, 170201 (2004)] is presented. Coulombic forces are mediated by a propagating electric field that obeys Gauss's law. A detailed description of the numerical implementation is presented, and the accuracy is calibrated by comparing the electrostatic force to that obtained from a standard Ewald summation. A parallelized implementation exhibits excellent scaling behavior on a contemporary low latency compute cluster, and therefore becomes an interesting alternative to existing algorithms for computing electrostatics in large-scale molecular dynamics. The approach opens new possibilities for the study of physical situations that are difficult or impossible to treat with standard reciprocal space methods, in particular nonperiodic boundary conditions and spatially heterogeneous dielectric environments in implicit solvent models. PMID- 17919009 TI - Theoretical investigation of the dissociation dynamics of vibrationally excited vinyl bromide on an ab initio potential-energy surface obtained using modified novelty sampling and feedforward neural networks. II. Numerical application of the method. AB - A previously reported method for conducting molecular dynamics simulations of gas phase chemical dynamics on ab initio potential-energy surfaces using modified novelty sampling and feedforward neural networks is applied to the investigation of the unimolecular dissociation of vinyl bromide. The neural network is fitted to a database comprising the MP4(SDQ) energies computed for 71 969 nuclear configurations using an extended basis set. Dissociation rate coefficients and branching ratios at an internal excitation energy of 6.44 eV for all six open reaction channels are reported. The distribution of vibrational energy in HBr formed in three-center dissociation is computed and found to be in excellent accord with experimental measurements. Computational requirements for the electronic structure calculations, neural network training, and trajectory calculations are given. The weight and bias matrices required for implementation of the neural network potential are made available through the Supplementary Material. PMID- 17919010 TI - Assessment of transition operator reference states in electron propagator calculations. AB - The transition operator method combined with second-order, self-energy corrections to the electron propagator (TOEP2) may be used to calculate valence and core-electron binding energies. This method is tested on a set of molecules to assess its predictive quality. For valence ionization energies, well known methods that include third-order terms achieve somewhat higher accuracy, but only with much higher demands for memory and arithmetic operations. Therefore, we propose the use of the TOEP2 method for the calculation of valence electron binding energies in large molecules where third-order methods are infeasible. For core-electron binding energies, TOEP2 results exhibit superior accuracy and efficiency and are relatively insensitive to the fractional occupation numbers that are assigned to the transition orbital. PMID- 17919011 TI - Electron affinity of (7)Li calculated with the inclusion of nuclear motion and relativistic corrections. AB - Explicitly correlated Gaussian functions have been used to perform very accurate variational calculations for the ground states of (7)Li and (7)Li(-). The nuclear motion has been explicitly included in the calculations (i.e., they have been done without assuming the Born-Oppenheimer (BO) approximation). An approach based on the analytical energy gradient calculated with respect to the Gaussian exponential parameters was employed. This led to a noticeable improvement of the previously determined variational upper bound to the nonrelativistic energy of Li(-). The Li energy obtained in the calculations matches those of the most accurate results obtained with Hylleraas functions. The finite-mass (non-BO) wave functions were used to calculate the alpha(2) relativistic corrections (alpha=1c). With those corrections and the alpha(3) and alpha(4) corrections taken from Pachucki and Komasa [J. Chem. Phys. 125, 204304 (2006)], the electron affinity (EA) of (7)Li was determined. It agrees very well with the most recent experimental EA. PMID- 17919012 TI - Formulation of magnetically perturbed time-dependent density functional theory. AB - A formulation of time-dependent density functional theory (TDDFT) in the presence of a static imaginary perturbation is derived. A perturbational approach is applied leading to corrections to various orders in the quantities of interest, namely, the excitation energies and transition densities. The perturbed TDDFT equations are relatively straightforward to derive but the resulting expressions are rather cumbersome. Simplifications of these equations are suggested. Both the simplified and full expressions are used to obtain equations for first- and second-order corrections to the excitation energy, the first-order correction to the transition density, and the corrections for both quantities to first-order in two different perturbations. This formulation, called magnetically perturbed TDDFT, details how conventional TDDFT calculations can be corrected to allow for the inclusion of a static magnetic field and/or spin-orbit coupling. PMID- 17919013 TI - Calculation of frequency-dependent hyperpolarizabilities using general coupled cluster models. AB - By exploiting the similarities between response theory and analytic derivative theory, we present a scheme for calculating frequency-dependent hyperpolarizabilities at the coupled-cluster level within the framework for analytic third derivatives. This has been implemented for arbitrary levels of coupled-cluster theory up to the full-configuration-interaction limit. An investigation of some small molecules shows that the inclusion of triple excitations is essential for an accurate description of hyperpolarizabilities. PMID- 17919014 TI - Time-dependent quantum Monte Carlo and the stochastic quantization. AB - We examine the relation between the recently proposed time-dependent quantum Monte Carlo (TDQMC) method and the principles of stochastic quantization. In both TDQMC and stochastic quantization, particle motion obeys stochastic guidance equations to preserve quantum equilibrium. In this way the probability density of the Monte Carlo particles corresponds to the modulus square of the many-body wave function at all times. However, in TDQMC, the motion of particles and guide waves occurs in physical space unlike in stochastic quantization where it occurs in configuration space. Hence, the practical calculation of time evolution of many body fully correlated quantum systems becomes feasible within the TDQMC methodology. We illustrate the TDQMC technique by calculating the symmetric and antisymmetric ground state of a model one-dimensional helium atom, and the time evolution of the dipole moment when the atom is irradiated by a strong ultrashort laser pulse. PMID- 17919015 TI - Molecular mechanics-valence bond method for planar conjugated hydrocarbon cations. AB - We present an extension of the molecular mechanics-valence bond (MMVB) hybrid method to study ground and excited states of planar conjugated hydrocarbon cations. Currently, accurate excited state calculations on these systems are limited to expensive ab initio studies of smaller systems: up to 15 active electrons in 16 pi orbitals with complete active space self-consistent field (CASSCF) theory using high symmetry. The new MMVB extension provides a faster, cheaper treatment to investigate larger cation systems with more than 24 active orbitals. Extension requires both new matrix elements and new parameters: In this paper we present both, for the limited planar case. The scheme is tested for the planar radical cations of benzene, naphthalene, anthracene, and phenanthrene. Calculated MMVB relative energies are in good agreement with CASSCF results for equilibrium geometries on the ground and first excited states, and conical intersections. PMID- 17919016 TI - Molecular properties determined from the relaxation of long-lived spin states. AB - The populations of long-lived spin states, in particular, populations of singlet states that are comprised of antisymmetric combinations of product states, |alpha(I)beta(S)> - |beta(I)alpha(S)>, are characterized by very long lifetimes because the dipole-dipole interaction between the two "active" spins I and S that are involved in such states is inoperative as a relaxation mechanism. The relaxation rate constants of long-lived (singlet) states are therefore determined by the chemical shift anisotropy (CSA) of the active spins and by dipole-dipole interactions with passive spins. For a pair of coupled spins, the singlet-state relaxation rate constants strongly depend on the magnitudes and orientations of the CSA tensors. The relaxation properties of long-lived states therefore reveal new information about molecular symmetry and structure and about spectral density functions that characterize the dynamic behavior. PMID- 17919017 TI - Ab initio energies of nonconducting crystals by systematic fragmentation. AB - A systematic method for approximating the ab initio electronic energy of molecules from the energies of molecular fragments has been adapted to estimate the total electronic energy of crystal lattices. The fragmentation method can be employed with any ab initio electronic structure method and allows optimization of the crystal structure based on ab initio gradients. The method is demonstrated on SiO(2) polymorphs using the Hartree-Fock approximation, second order Moller Plesset perturbation theory, and the quadratic configuration interaction method with single and double excitations and triple excitations added perturbatively . PMID- 17919018 TI - Photoabsorption and photofragmentation of isolated cationic silver cluster tryptophan hybrid systems. AB - We present a theoretical study of the size and structure selective absorption properties of cationic silver cluster-tryptophan Trp-Ag(n)(+) (n = 2-5,9) hybrid systems supported by photofragmentation experiments. Our time-dependent density functional theory calculations provide insight into the nature of excitations in interacting nanoparticle-biomolecule subunits and allow to determine characteristic spectral features as fingerprints of two different classes of structures: charge solvated and zwitterionic. Moreover, different types of charge transfer transitions have been identified. Charge transfer from pi system of tryptophan to silver cluster occurs for charge solvated structures while charge transfer from silver to the NH(3) (+) group takes place for zwitterionic structures. This has been confirmed by experimentally measured photofragmentation channels and molecular dynamics simulations. Our findings provide fundamental insight into the structure- and size-dependent mechanism responsible for the enhanced absorption and emission in nanoparticle-biomolecular hybrid systems. PMID- 17919019 TI - Effective interaction energy of water dimer at room temperature: an experimental and theoretical study. AB - Buffer-gas pressure broadening for the nu(1)+nu(3) band of H(2)O at 1.34-1.44 mum for a variety of buffer gases was investigated at room temperature using continuous-wave cavity ring-down spectroscopy. The effective interaction energy of water dimer under room temperature conditions was evaluated from the pressure broadening coefficients for rare gases using Permenter-Seaver's relation. Monte Carlo simulations were performed using ab initio molecular orbital calculations to evaluate the interaction energies for the water dimer at 300 K. In this theoretical calculation, the orientations of the two water molecules were statistically treated. PMID- 17919020 TI - Electron impact ionization of water-doped superfluid helium nanodroplets: observation of He(H(2)O)(n)(+) clusters. AB - Electron impact mass spectra have been recorded for helium nanodroplets containing water clusters. In addition to identification of both H(+)(H(2)O)(n) and (H(2)O)(n)(+) ions in the gas phase, additional peaks are observed which are assigned to He(H(2)O)(n)(+) clusters for up to n=27. No clusters are detected with more than one helium atom attached. The interpretation of these findings is that quenching of (H(2)O)(n)(+) by the surrounding helium can cool the cluster to the point where not only is fragmentation to H(+)(H(2)O)(m) (where m < or = n-1) avoided, but also, in some cases, a helium atom can remain attached to the cluster ion as it escapes into the gas phase. Ab initio calculations suggest that the first step after ionization is the rapid formation of distinct H(3)O(+) and OH units within the (H(2)O)(n)(+) cluster. To explain the formation and survival of He(H(2)O)(n)(+) clusters through to detection, the H(3)O(+) is assumed to be located at the surface of the cluster with a dangling O-H bond to which a single helium atom can attach via a charge-induced dipole interaction. This study suggests that, like H(+)(H(2)O)(n) ions, the preferential location for the positive charge in large (H(2)O)(n)(+) clusters is on the surface rather than as a solvated ion in the interior of the cluster. PMID- 17919021 TI - Local mode behavior in H(2)Te. AB - A high-resolution spectrum of hydrogen telluride (H(2)Te) was recorded in the 4050-7000 cm(-1) region. Two bands could be observed at 4900 and 5980 cm(-1) and were assigned as the (20(+/-),1) <-- (00(+),0) and (30(+/-),0) <-- (00(+),0) bands, respectively. Rotational transitions of the two bands were assigned for the most abundant H(2) (130)Te and H(2) (128)Te isotopic species. Line position analyses were carried out to investigate a possible local mode behavior. For the first time we found in H(2)Te strong experimental evidence for such a behavior for the higher-lying band as its two upper vibrational states are only 0.027 and 0.032 cm(-1) apart for the H(2) (130)Te and H(2) (128)Te isotopic species, respectively. PMID- 17919022 TI - Inelastic collisions in molecular nitrogen at low temperature (2 < or = T < or = 50 K). AB - Theory and experiment are combined in a novel approach aimed at establishing a set of two-body state-to-state rates for elementary processes ij --> lm in low temperature N(2):N(2) collisions involving the rotational states i,j,l,m. First, a set of 148 collision cross sections is calculated as a function of the collision energy at the converged close-coupled level via the MOLSCAT code, using a recent potential energy surface for N(2)-N(2). Then, the corresponding rates for the range of 2 < or = T < or = 50 K are derived from the cross sections. The link between theory and experiment, aimed at assessing the calculated rates, is a master equation which accounts for the time evolution of rotational populations in a reference volume of gas in terms of the collision rates. In the experiment, the evolution of rotational populations is measured by Raman spectroscopy in a tiny reference volume (approximately 2 x 10(-3) mm(3)) of N(2) traveling along the axis of a supersonic jet. The calculated collisional rates are assessed experimentally in the range of 4 < or = T < or = 35 K by means of the master equation, and then are scaled by averaging over a large set of experimental data. The scaled rates account accurately for the evolution of the rotational populations measured in a wide range of conditions. Accuracy of 10% is estimated for the main scaled rates. PMID- 17919023 TI - On the limitations of adiabatic population transfer between molecular electronic states induced by intense femtosecond laser pulses. AB - The possibility to perform a stimulated Raman adiabatic passage process in molecules on the ultrafast time scale is investigated theoretically. Motivated by recent experiments, the mid R:B<--mid R:X electronic transitions in molecular iodine are studied as a prototype example with the goal to selectively induce a population transfer employing two intense and time-delayed ultrashort laser pulses and different coupling schemes. For the purpose of interpretation, the coupled multilevel vibronic problem is reduced to a quasi-three-level system by averaging over the vibrational degree of freedom. It is shown that the vibrational dynamics becomes essential at high field intensities. Considering a 2 dimensional parameter space (intensity and delay time of the femtosecond laser pulses), a strong-field control landscape is constructed. PMID- 17919024 TI - Resonant two-photon ionization and ab initio conformational analysis of haloethyl benzenes (PhCH(2)CH(2)X,X=Cl,F). AB - The S(1) <-- S(0) transitions of the gaseous (2-fluoroethyl)-benzene (FEB) and (2 chloroethyl)-benzene (CEB) have been investigated using a combination of two color resonant two-photon ionization and UV-UV hole burning spectroscopy. Both anti and gauche conformers have been identified on the basis of rotational band contour analysis supported by ab initio calculations on the ground and electronically excited states. The gauche origin band of FEB at 37,673 cm(-1) is redshifted 50 cm(-1) relative to the corresponding anti origin, while CEB origin bands overlap at 37,646 cm(-1). Relative conformational stability and populations in the jet have been estimated for both molecules, based on the intensity ratio of S(1) <-- S(0) band origin transitions. These are compared with a range of related molecules with the structural motif PhCH(2)CH(2)X (X=CH(3),CH(2)CH(3),NH(2),OH,COOH,CCH,CN). Theory and experimental results for FEB and CEB show repulsive interactions between the halogen substituents and the pi cloud of the phenyl rings destabilizing the gauche conformers, but the preference for the anti conformers is relatively modest. The gauche conformer origins show very different hybrid character: FEB is largely b type, while CEB is an ac hybrid in keeping with theoretically computed TM "rotations" (theta(elec)) of -7 degrees and -56 degrees , respectively. This difference is attributed largely to rotation of the side chain in opposite directions about the C(1)C(alpha) bond. Spectra of FEB(H(2)O) and CEB(H(2)O) single water clusters show evidence of an anti conformation in the host molecule. PMID- 17919025 TI - Moments of hyper-Rayleigh spectra of selected rare gas mixtures. AB - In this work we have analyzed spectral moments characterizing properties of the collisionally hyper-Rayleigh scattered light. This is a supplementary study undertaken in order to complete the series of our previously published papers on the collisional hyper-Rayleigh scattering spectral profiles. In order to evaluate the moments we have extended the theory so that it could embrace the (hyper)polarizabilities of higher rank. Using the expressions developed on the grounds of the theoretical principles and applying appropriate computational methods with ab initio hyperpolarizability values as an input, we have obtained desirable moment values for three diatomic noble gas systems: HeNe, HeAr, and NeAr, at several temperature points. The semiclassical and the quantum treatments have been taken into account, and the moments were calculated both from the sum rule method as well as from the spectral profiles. The results were compared and discussed. PMID- 17919026 TI - The radical character of the acenes: a density matrix renormalization group study. AB - We present a detailed investigation of the acene series using high-level wave function theory. Our ab initio density matrix renormalization group algorithm has enabled us to carry out complete active space calculations on the acenes from napthalene to dodecacene correlating the full pi-valence space. While we find that the ground state is a singlet for all chain lengths, examination of several measures of radical character, including the natural orbitals, effective number of unpaired electrons, and various correlation functions, suggests that the longer acene ground states are polyradical in nature. PMID- 17919027 TI - Intermetallic compounds of the heaviest elements and their homologs: the electronic structure and bonding of MM', where M=Ge, Sn, Pb, and element 114, and M'=Ni, Pd, Pt, Cu, Ag, Au, Sn, Pb, and element 114. AB - Fully relativistic (four-component) density-functional theory calculations were performed for intermetallic dimers MM', where M=Ge, Sn, Pb, and element 114, and MM'=group 10 elements (Ni, Pd, and Pt) and group 11 elements (Cu, Ag, and Au). PbM and 114M, where M are group 14 elements, were also considered. The results have shown that trends in spectroscopic properties-atomization energies D(e), vibrational frequencies omega(e), and bond lengths R(e), as a function of MM', are similar for compounds of Ge, Sn, Pb, and element 114, except for D(e) of PbNi and 114Ni. They were shown to be determined by trends in the energies and space distribution of the valence ns(MM')atomic orbitals (AOs). According to the results, element 114 should form the weakest bonding with Ni and Ag, while the strongest with Pt due to the largest involvement of the 5d(Pt) AOs. In turn, trends in the spectroscopic properties of MM' as a function of M were shown to be determined by the behavior of the np(1/2)(M) AOs. Overall, D(e) of the element 114 dimers are about 1 eV smaller and R(e) are about 0.2 a.u. larger than those of the corresponding Pb compounds. Such a decrease in bonding of the element 114 dimers is caused by the large SO splitting of the 7p orbitals and a decreasing contribution of the relativistically stabilized 7p(1/2)(114) AO. On the basis of the calculated D(e) for the dimers, adsorption enthalpies of element 114 on the corresponding metal surfaces were estimated: They were shown to be about 100-150 kJ/mol smaller than those of Pb. PMID- 17919028 TI - Control of concerted two bond versus single bond dissociation in CH(3)Co(CO)(4) via an intermediate state using pump-dump laser pulses. AB - Wavepacket propagations on ab initio multiconfigurational two-dimensional potential energy surfaces for CH(3)Co(CO)(4) indicate that after irradiation to the lowest first and second electronic excited states, concerted dissociation of CH(3) and the axial CO ligand takes place. We employ a pump-dump sequence of pulses with appropriate frequencies and time delays to achieve the selective breakage of a single bond by controlling the dissociation angle. The pump and dump pulse sequence exploits the unbound surface where dissociation occurs in a counterintuitive fashion; stretching of one bond in an intermediate state enhances the single dissociation of the other bond. PMID- 17919029 TI - Fourier transform microwave spectroscopy of the isocyanomethyl radical, CH(2)NC. AB - The pure rotational spectrum of the isocyanomethyl radical, CH(2)NC, was measured for the first time by using a Fourier transform microwave spectrometer. The molecule was produced by a discharge of isocyanomethane, CH(3)NC, diluted in Ar or Ne. The spectral lines due to the N=1-0 and 2-1 transitions were recorded near 22 and 44 GHz, respectively. The observed spectrum showed a complicated fine and hyperfine structure because of the same order of interaction energies. Among the 39 spectral lines detected and assigned, the transitions with K(a)=1 show no hyperfine splitting due to the hydrogen nuclei, suggesting planarity for the molecule. Molecular constants such as rotational and spin-rotational parameters including centrifugal effects and hyperfine coupling constants due to both the nitrogen and the hydrogen nuclei were accurately determined. The structure and the astronomical implications of the molecule are discussed. PMID- 17919030 TI - Comprehensive characterization of the photodissociation pathways of protonated tryptophan. AB - The photofragmentation of protonated tryptophan has been investigated in a unique experimental setup, in which ion and neutral issued from the photofragmentation are detected in coincidence, in time and in position. From these data are extracted the kinetic energy, the number of neutral fragments associated with an ion, their masses, and the order of the fragmentation steps. Moreover, the fragmentation time scale ranging from tens of nanoseconds to milliseconds is obtained. From all these data, a comprehensive fragmentation mechanism is proposed. PMID- 17919031 TI - Matrix isolation spectroscopy and molecular dynamics simulations for 2,7,12,17 tetra-tert-butylporphycene in argon and xenon. AB - Electronic absorption spectra of 2,7,12,17-tetra-tert-butylporphycene (TTPC) have been recorded in low-temperature argon and xenon matrices for various deposition conditions. In the region of the S(0)-S(1) electronic transition, the spectra of TTPC in argon reveal a rich site structure, characterized by a series of more than 30 absorption peaks. Studies of the temperature dependence of the electronic spectra in solid argon demonstrated remarkable spectral changes, resulting in the broadening of all bands with increasing temperature. These temperature-induced spectral changes are, to a large degree, reversible, so lowering of temperature is accompanied by the recovery of the original fine-line spectrum. The absorption spectra in xenon reveal broad bands, on which a rich pattern of lines becomes superimposed at low temperatures. Trapping site distribution and the structure of the microenvironment around the TTPC chromophore, embedded in argon and xenon hosts, have been analyzed using molecular dynamics (MD) simulations. The MD results show that the trapping of TTPC in rare-gas solids is influenced by favorable embedding of the bulky tert-butyl groups inside the matrix cage. The crucial role of the tert-butyl groups for the thermodynamics and kinetics of matrix deposition is demonstrated by comparing the results with those obtained for the parent, unsubstituted porphycene. PMID- 17919032 TI - Oxygen K-edge fine structures of water by x-ray Raman scattering spectroscopy under pressure conditions. AB - Fine structure of the oxygen K edge was investigated for water at ambient pressure, 0.16, 0.21, 0.27, 0.47, and 0.60 GPa using x-ray Raman scattering spectroscopy (XRS). Similarity in near-edge structures at 0.16 and 0.60 GPa suggests little difference in the electronic state of oxygen in the low-pressure and high-pressure forms of water. Yet, we observed significant variation of preedge structure of the XRS spectra with compression. The intensity of the preedge peak at 535.7 eV has a minimal value at around 0.3 GPa, indicating that the number of hydrogen bonding increases first and then decreases as a function of pressure. PMID- 17919033 TI - Simulated thermal decomposition and detonation of nitrogen cubane by molecular dynamics. AB - We present simulations of a model molecular solid of nitrogen cubane subject to thermal agitation and mechanical shock. A new approach, a reactive state summation potential, has been used to model nitrogen cubane dissociation. At elevated temperatures, the system decomposes to N(2) mixed with a small amount of oligomeric nitrogen. When subject to shock loading the system detonates above some critical threshold after which a shock front is self-sustained by the energy release from chemical reactions at a constant intrinsic speed. This is the first example of a fully three-dimensional atomic simulation of a chemically-sustained detonation. The spatial confinement of the shock front results in longer chain intermediates than in the case of thermal decomposition, suggesting that shock intermediates can be structurally very different from the same material subject to comparable temperatures and pressures. PMID- 17919034 TI - Topological building blocks of hydrogen bond network in water. AB - Basic three-dimensional units of the network, called fragments, are introduced to characterize the hydrogen bond (HB) network structure of water. Topological differences among normal liquid water, water at low temperature, and water under high pressure are elucidated by their fragment statistics. Water at low temperature has almost defect-free network and is filled with stable fragments with small distortion. It is found that there exists a certain way on how fragments mutually aggregate. Well-formed aggregates heterogeneously constitute very stable network structures. HB network rearrangements occur scarcely inside these aggregated domains but take place in their surface areas. The heterogeneity of HB structure and rearrangement in water is thus explained in terms of the fragment structure and its rearrangements. The fragment analysis thus elucidates the intermediate-range order in water HB network. PMID- 17919035 TI - Inelastic neutron scattering study of hydrogen in d(8)-THFD(2)O ice clathrate. AB - In situ neutron inelastic scattering experiments on hydrogen adsorbed into a fully deutrated tetrahydrofuran-water ice clathrate show that the adsorbed hydrogen has three rotational excitations (transitions between J=0 and 1 states) at approximately 14 meV in both energy gain and loss. These transitions could be unequivocally assigned since there was residual orthohydrogen at low temperatures (slow conversion to the ground state) resulting in an observable J=1-->0 transition at 5 K (kT=0.48 meV). A doublet in neutron energy loss at approximately 28.5 meV is interpreted as J=1-->2 transitions. In addition to the transitions between rotational states, there are a series of peaks that arise from transitions between center-of-mass translational quantum states of the confined hydrogen molecule. A band at approximately 9 meV can be unequivocally interpreted as a transition between translational states, while broad features at 20, 25, 35, and 50-60 meV are also interpreted to as transitions between translational quantum states. A detailed comparison is made with a recent five dimensional quantum treatment of hydrogen in the smaller dodecahedral cage in the SII ice-clathrate structure. Although there is broad agreement regarding the features such as the splitting of the J=1 degeneracy, the magnitude of the external potential is overestimated. The numerous transitions between translational states predicted by this model are in poor agreement with the experimental data. Comparisons are also made with three simple exactly solved models, namely, a particle in a box, a particle in a sphere, and a particle on the surface of a sphere. Again, there are too many predicted features by the first two models, but there is reasonable agreement with the particle on a sphere model. This is consistent with published quantum chemistry results for hydrogen in the dodecahedral 5(12) cage, where the center of the cage is found to be energetically unfavorable, resulting in a shell-like confinement for the hydrogen molecule wave function. These results demonstrate that translational quantum effects are very significant and a classical treatment of the hydrogen molecule dynamics is inappropriate under such conditions. PMID- 17919036 TI - Atomistic simulation of discotic liquid crystals: transition from isotropic to columnar phase example. AB - Molecular dynamics simulations at atomistic level have been performed on a metal porphyrazine complex. Starting from an isotropic state, the system was cooled until transition from isotropic to columnar phase was observed; no nematic phase was encountered. Many tools were utilized to follow the system evolution: order parameter, g(r), g(||)(r(||)), g(c)(r(||)), g(perpendicular)(r(perpendicular)), g(2)(r), also density and energy changes. Very long runs were required to get reliable results, times greater than 40 ns of simulation. The structure of columnar phase was analyzed and the organization of molecules in the columns was investigated, along with the role of conformation of side chains. We found that in columnar phase the molecules are tilted versus the column axis and the conformation of side chains changes during the phase transition to allow this kind of organization; moreover the direction of columns axes is different from that of the director. PMID- 17919037 TI - Molecular simulations of droplet coalescence in oil/water/surfactant systems. AB - We report a molecular simulation study of the mechanism by which droplets covered with a surfactant monolayer coalesce. We study a model system where the rate limiting step in coalescence is the rupture of the surfactant film. Our simulations allow us to focus on the stages at the core of the coalescence process: the initial rupture of the two surfactant monolayers, the rearrangement of the surfactant molecules to form a channel connecting the two droplets, and the expansion of the radius of the resulting channel. For our numerical study, we made use of the dissipative particle dynamics method. We used a coarse-grained description of the oil, water, and surfactant molecules. The rupture of the surfactant film is a rare event on the molecular time scale. To enhance the sampling of the rupture of the surfactant film, we used forward flux sampling (FFS). FFS not only allows us to estimate coalescence rates, it also provides insight into the molecular structure and free energy of the "transition" state. For an oil-water-oil film without surfactant, the rupture rate decreases exponentially with increasing film thickness. The critical state is different in thin and thick films: Thin films break following a large enough thickness fluctuation. Thicker films break only after a sufficiently large hole fluctuation they can heal. Next, we designed surfactant molecules with positive, zero, and negative natural curvatures. For a water film between two surfactant-covered oil droplets, the rupture rate is highest when the surfactant has a negative natural curvature, lowest when it has zero natural curvature, and lying in between when it has a positive natural curvature. This nonmonotonic variation with curvature stems from two effects: First, the surfactants with a large absolute value of the natural curvature have lower interfacial tension and bending rigidity. This promotes the interfacial fluctuations required to nucleate a channel. Second, the sign of the natural curvature determines whether there is a critical channel radius at which the channel free energy has a maximum. The latter is in agreement with the hole-nucleation theory of Kabalnov and Wennerstrom [Langmuir 12, 276 (1996)]. Our simulations seriously overestimate the relative stability of surfactant free emulsions. We argue that this is due to the fact that our model does not allow for nanobubble formation and capillary evaporation-processes that are presumably of key importance in the coalescence of surfactant-free emulsions. PMID- 17919038 TI - Grand canonical Monte Carlo simulation study of capillary condensation between nanoparticles. AB - Capillary condensation at the nanoscale differs from condensation in the bulk phase, because it is a strong function of surface geometry and gas-surface interactions. Here, the effects of geometry on the thermodynamics of capillary condensation at the neck region between nanoparticles are investigated via a grand canonical Monte Carlo simulation using a two-dimensional lattice gas model. The microscopic details of the meniscus formation on various surface geometries are examined and compared with results of classical macromolecular theory, the Kelvin equation. We assume that the system is composed of a lattice gas and the surfaces of two particles are approximated by various shapes. The system is modeled on the basis of the molecular properties of the particle surface and lattice gas in our system corresponding to titania nanoparticles and tetraethoxy orthosilicate molecules, respectively. This system was chosen in order to reasonably emulate our previous experimental results for capillary condensation on nanoparticle surfaces. Qualitatively, our simulation results show that the specific geometry in the capillary zone, the surface-surface distance, and the saturation ratio are important for determining the onset and broadening of the liquid meniscus. The meniscus height increases continuously as the saturation ratio increases and the meniscus broadens faster above the saturation ratio of 0.90. The change of the radius of curvature of the particle surface affects the dimensions of the capillary zone, which drives more condensation in narrow zones and less condensation in wide zones. The increase of surface-surface distance results in the decrease of the meniscus height or even the disappearance of the meniscus entirely at lower saturation ratios. These effects are significant at the nanoscale and must be carefully considered in order to develop predictive relationships for meniscus height as a function of saturation conditions. PMID- 17919039 TI - In-plane structure and ordering at liquid sodium surfaces and interfaces from ab initio molecular dynamics. AB - Atoms at liquid metal surfaces are known to form layers parallel to the surface. We analyze the two-dimensional arrangement of atoms within such layers at the surface of liquid sodium using ab initio molecular dynamics (MD) simulations based on a full version of density functional theory. Nearest neighbor distributions at the surface indicate mostly fivefold coordination, though there are noticeable fractions of fourfold and sixfold coordinated atoms. Bond angle distributions suggest a movement toward the angles corresponding to a sixfold coordinated hexagonal arrangement of the atoms as the temperature is decreased towards the solidification point. We rationalize these results with a distorted hexagonal model at the surface, showing a mixture of regions of five- and sixfold coordination. The liquid surface results are compared with classical MD simulations of the liquid surface, with similar effects appearing, and with ab initio MD simulations for a model solid-liquid interface, where a pronounced shift towards hexagonal ordering is observed as the temperature is lowered. PMID- 17919040 TI - Tuning ferroelectricity of niobium clusters by oxygen chemisorption. AB - The chemisorption of atomic oxygen on Nb(N) (N=2-16) has been investigated by the scalar relativistic all-electron density-functional calculations with emphasis on its effect on the ferroelectricity of Nb(N). We have shown that the binding of O atom to niobium clusters is site- and size-dependent, for which the bridge-site doping is preferred in the smaller size range from N=2 to 8 and the threefold hollow site one for the larger Nb(N) with 9 < or = N < or = 16. Though the geometrical structures of Nb(N) are modified slightly when doped with an oxygen atom, their ferroelectric properties vary considerably, depending on cluster size and the O adsorption sites, which is mainly caused by the charge transfer between the oxygen atom and niobium clusters. The addition of oxygen can enhance the ferroelectricity of Nb(N) with moderate and essentially zero moments while induce only small dipole change for those Nb(N) with large moments. Thus, the big fluctuation of the Nb(N)'s dipole moments with size is greatly suppressed by the O doping, indicating that one might expect to tune the size-dependent ferroelectricity of Nb(N) by the chemical decoration. PMID- 17919041 TI - Electronic properties of adsorbates on GaAs(001)-c(2x8)/(2x4). AB - A systematic experimental and theoretical study was performed to determine the causes of oxide-induced Fermi level pinning and unpinning on GaAs(001)-c(2 x 8)/(2 x 4). Scanning tunneling spectroscopy (STS) and density functional theory (DFT) were used to study four different adsorbates' (O(2), In(2)O, Ga(2)O, and SiO) bonding to the GaAs(001)-c(2 x 8)/(2 x 4) surface. The STS results revealed that out of the four adsorbates studied, only one left the Fermi level unpinned, Ga(2)O. DFT calculations were used to elucidate the causes of the Fermi level pinning. Two distinct pinning mechanisms were identified: direct (adsorbate induced states in the band gap region) and indirect pinnings (generation of undimerized As atoms). For O(2) dissociative chemisorption onto GaAs(001)-c(2 x 8)/(2 x 4), the Fermi level pinning was only indirect, while direct Fermi level pinning was observed when In(2)O was deposited on GaAs(001)-c(2 x 8)/(2 x 4). In the case of SiO on GaAs(001)-c(2 x 8)/(2 x 4), the Fermi level pinning was a combination of the two mechanisms. PMID- 17919042 TI - Low energy electron diffraction and low energy electron microscopy microspot I/V analysis of the (4 x 4)O structure on Ag(111): surface oxide or reconstruction? AB - A low energy electron diffraction (LEED) I/V analysis was performed of the (4 x 4) oxygen structure on Ag(111). Two data sets were used, one recorded with a conventional LEED system and a second with a low energy electron microscope (LEEM). The data sets agree well with each other, demonstrating that I/V structure analyses can be performed with the same quality with LEEM as with conventional LEED. The structure obtained confirms the recently proposed model that involves a reconstruction of the Ag(111) surface. Previous models based on a thin layer of Ag(2)O that had been accepted for more than 30 years are disproved. The reconstruction model contains two units of six triangularly arranged Ag atoms and a stacking fault in one half of the unit cell. The six O atoms per unit cell occupy sites in the trenches between the Ag(6) triangles. Small lateral displacements of the Ag atoms lift the mirror symmetry of the structure, leading to two nonequivalent groups of O atoms. The atoms of both groups are located approximately 0.5 Angstrom below the top Ag layer, on fourfold positions with respect to the top layer Ag atoms. Ag-O distances between 2.05 and 2.3 Angstrom are found. The oxygen atoms exhibit large static or dynamic displacements of up to 0.3 Angstrom at 300 K. PMID- 17919043 TI - Adsorption and charge transfer dynamics of bi-isonicotinic acid on Au(111). AB - The interaction of bi-isonicotinic acid (4,4(')-dicarboxy-2,2(')-bipyridine) with the Au(111) surface has been investigated using electron spectroscopic techniques. Near edge x-ray absorption fine structure (NEXAFS) spectra show that monolayers of the molecule lie flat to the surface and also reveal that the monolayer is sensitive to the preparation conditions employed. Core level x-ray photoelectron spectroscopy (XPS) shows that the adsorbed molecule does not undergo deprotonation upon adsorption. The "core-hole clock" implementation of resonant photoemission has been used to probe the coupling between molecule and substrate. This technique has revealed the possibility of ultrafast backtransfer from the substrate into the molecule upon resonant excitation of a N 1s core level electron. This is supported by a NEXAFS and XPS investigation of energy level alignments in the system. PMID- 17919044 TI - Molecular dynamics simulations of signal transmission through a glycine peptide chain. AB - The injection of finite duration vibrational signals encoding information into a biomolecular wire of the polypeptide glycine1000 is investigated theoretically using molecular dynamics simulations and digital signal processing techniques. We demonstrate that the amplitude modulated signal applied to one of the C-N bonds of the molecule transmits in the two directions through the long polypeptide molecule, which is connected to gold clusters at each of its ends. A decay of the signal propagation speed is observed along with intensity decay. On the other hand, the molecular dynamics simulations show that signal transmission is completely achievable at room temperature, thus realistic transmission of signals through linear molecules can be performed. PMID- 17919045 TI - Chemiluminescent detection of neutral gaseous radicals. AB - This paper presents a systematic approach to the development of novel solid-state chemical sensors on the basis of heterogeneous chemiluminescence. The method is applicable for the identification and measurements of concentration of H, O, and other gaseous chemical radicals, where utilization of standard techniques is difficult. The luminescence is invoked during Eley-Rideal recombination of the radicals in question on the surface of the sensor core. A technique is discussed to separate the contributions of Eley-Rideal and Langmuir-Hinshelwood mechanisms, and to select appropriate materials for the sensor emitter fabrication. Typical sensor characteristics include sensitivity of 10(5) cm(-3), working gas pressures of 10(-8) - 10(1) Torr, and measurement time approximately 1 s. PMID- 17919046 TI - Two-component heterogeneous nucleation kinetics and an application to Mars. AB - We develop a two-component heterogeneous nucleation model that includes exact calculation of the Stauffer-type [D. Stauffer, J. Aerosol Sci. 7, 319 (1976)] steady-state kinetic prefactor using the correct heterogeneous Zeldovich factor for a heterogeneous two-component system. The model, and a simplified version of it, is tested by comparing its predictions to experimental data for water-n propanol nucleating on silver particles. The model is then applied to water carbon dioxide system in Martian conditions, which has not been modeled before. Using the ideal mixture assumption, the model shows theoretical possibilities for two-component nucleation adjacent to the initial stages of one-component water nucleation, especially with small water vapor amounts. The numbers of carbon dioxide molecules in the critical cluster are small in the case of large water amounts (up to 300 ppm) in the gas phase, but larger when there is very little water vapor (1 ppm). PMID- 17919047 TI - Quantum dynamical treatment of inelastic scattering of atoms at a surface at finite temperature: the random phase thermal wave function approach. AB - We present quantum dynamical calculations for the inelastic scattering of atoms at a nonrigid surface at finite temperature. The surface degrees of freedom are discretized and treated in a multiconfigurational wave function picture. The thermal averaging is carried out with the random phase thermal wave function approach. We show that it is sufficient to restrict the random phases to the intermediate basis of single particle functions, discuss the convergence of the method with the number of configurations and realizations, and analyze the flow of energy between different parts of the system for a range of temperatures between 4 and 500 K. PMID- 17919048 TI - A single particle model to simulate the dynamics of entangled polymer melts. AB - We present a computer simulation model of polymer melts representing each chain as one single particle. Besides the position coordinate of each particle, we introduce a parameter n(ij) for each pair of particles i and j within a specified distance from each other. These numbers, called entanglement numbers, describe the deviation of the system of ignored coordinates from its equilibrium state for the given configuration of the centers of mass of the polymers. The deviations of the entanglement numbers from their equilibrium values give rise to transient forces, which, together with the conservative forces derived from the potential of mean force, govern the displacements of the particles. We have applied our model to a melt of C(800)H(1602) chains at 450 K and have found good agreement with experiments and more detailed simulations. Properties addressed in this paper are radial distribution functions, dynamic structure factors, and linear as well as nonlinear rheological properties. PMID- 17919049 TI - Development of the energy flow in light-harvesting dendrimers. AB - Modeling the multistep flow of energy in light-harvesting dendrimers presents a considerable challenge. Recent studies have introduced an operator approach based on a matrix representation of the connectivity between constituent chromophores. Following a review of the theory, detailed applications are now shown to exhibit the time development of the core excitation following pulsed laser irradiation and the steady-state behavior that can be expected under conditions of constant illumination. It is also shown how energy capture by whole dendrimers can be analytically related to chromophore pair-transfer properties and, in particular, the spectroscopic gradient toward the core. Indicative calculations also illustrate the consequences of tertiary folding. In each respect, the model affords opportunities to derive new, physically meaningful information on the photophysical and structural features of dendrimeric systems. PMID- 17919050 TI - Configurational probabilities for symmetric dimers on a lattice: an analytical approximation with exact limits at low and high densities. AB - A new approach is developed for lattice density functional theory of interacting symmetric dimers at high temperatures. Equations of equilibrium for two dimensional square and three-dimensional cubic lattices are derived for the complete set of configurations in the first three shells around the central dimer, and rules of truncation for higher shells are based on exact results from the mathematical theory of domino tilings. This provides exact limits for both low and high densities. The new model predicts contributions of particular configurations which are in agreement with Monte Carlo simulations over the whole range of densities, including agreement with pocket Monte Carlo simulations at high densities. PMID- 17919051 TI - Spectra of Husimi cacti: exact results and applications. AB - Starting from exact relations for finite Husimi cacti we determine their complete spectra to very high accuracy. The Husimi cacti are dual structures to the dendrimers but, distinct from these, contain loops. Our solution makes use of a judicious analysis of the normal modes. Although close to those of dendrimers, the spectra of Husimi cacti differ. From the wealth of applications for measurable quantities which depend only on the spectra, we display for Husimi cacti the behavior of the fluorescence depolarization under quasiresonant Forster energy transfer. PMID- 17919052 TI - Soft particle model for block copolymers. AB - A soft particle model for diblock (AB) copolymer melts is proposed. Each molecule is mapped onto two soft spheres built by Gaussian A- and B-monomer distributions. An approximate analytical expression for the joint distribution function for the distance between both spheres and their radii of gyration is derived, which determines the entropic contribution to the intramolecular free energy. Adding a mean-field expression for the intermolecular interactions, we obtain the total free energy of the system. Based on this free energy, Monte Carlo simulations are carried out to study the kinetics of microphase ordering in the bulk and its effect on molecular diffusion. This is followed by an analysis of thin films, with emphasis on pattern transfer from walls with a periodic structure. It is shown that the level of coarse graining in the soft particle model is suitable to describe structural and kinetic properties of copolymers on mesoscopic scales. PMID- 17919053 TI - First-principles investigation on the geometry and electronic structure of the three-dimensional cuboidal C(60) polymer. AB - The structural stability and electronic properties of the recently characterized three-dimensional (3D) cuboid-shaped C(60) polymer are studied using periodic ab initio density functional methods. It is shown that the experimentally observed structure is metastable and not fully relaxed from the high pressure state. A second polymorph, which is more stable than the experimental structure, is identified from the calculations. This new structure differs from the observed structure in the number of fourfold-coordinated atoms per C(60) molecule. Both structures are found to be metallic with bulk moduli only about one-third that of diamond. The cuboidal C(60) is not the long sought after superhard 3D carbon polymer; however, the two polymorphs studied here reveal unusual electronic band structures that might suggest interesting electronic properties. PMID- 17919054 TI - Diffusive dynamics on multidimensional rough free energy surfaces. AB - The dynamics of processes relevant to chemistry and biophysics on rough free energy landscapes is investigated using a recently developed algorithm to solve the Smoluchowski equation. Two different processes are considered: ligand rebinding in MbCO and protein folding. For the rebinding dynamics of carbon monoxide (CO) to native myoglobin (Mb) from locations around the active site, the two-dimensional free energy surface (FES) is constructed using extensive molecular dynamics simulations. The surface describes the minima in the A state (bound MbCO), CO in the distal pocket and in the Xe4 pocket, and the transitions between these states and allows to study the diffusion of CO in detail. For the folding dynamics of protein G, a previously determined two-dimensional FES was available. To follow the diffusive dynamics on these rough free energy surfaces, the Smoluchowski equation is solved using the recently developed hierarchical discrete approximation method. From the relaxation of the initial nonequilibrium distribution, experimentally accessible quantities such as the rebinding time for CO or the folding time for protein G can be calculated. It is found that the free energy barrier for CO in the Xe4 pocket and in the distal pocket (B state) closer to the heme iron is approximately 6 kcal/mol which is considerably larger than the inner barrier which separates the bound state and the B state. For the folding of protein G, a barrier of approximately 10 kcal/mol between the unfolded and the folded state is consistent with folding times of the order of milliseconds. PMID- 17919056 TI - Asymmetric band profile of the Soret band of deoxymyoglobin is caused by electronic and vibronic perturbations of the heme group rather than by a doming deformation. AB - We measured the Soret band of deoxymyoglobin (deoxyMb), myoglobin cyanide (MbCN), and aquo-metmyoglobin (all from horse heart) with absorption and circular dichroism (CD) spectroscopies. A clear non-coincidence was observed between the absorption and CD profiles of deoxyMb and MbCN, with the CD profiles red- and blueshifted with respect to the absorption band position, respectively. On the contrary, the CD and absorption profiles of aquametMb were nearly identical. The observed noncoincidence indicates a splitting of the excited B state due to heme protein interactions. CD and absorption profiles of deoxyMb and MbCN were self consistently analyzed by employing a perturbation approach for weak vibronic coupling as well as the relative intensities and depolarization ratios of seven bands in the respective resonance Raman spectra measured with B-band excitation. The respective B(y) component was found to dominate the observed Cotton effect of both myoglobin derivatives. The different signs of the noncoincidences between CD and absorption bands observed for deoxyMb and MbCN are due to different signs of the respective matrix elements of A(1g) electronic interstate coupling, which reflects an imbalance of Gouterman's 50:50 states. The splitting of the B band reflects contributions from electronic and vibronic perturbations of B(1g) symmetry. The results of our analysis suggest that the broad and asymmetric absorption band of deoxyMb results from this band splitting rather than from its dependence on heme doming. Thus, we are able to explain recent findings that the temperature dependences of CO rebinding to myoglobin and the Soret band profile are uncorrelated[Ormos et al., Proc. Natl. Acad. Sci U.S.A. 95, 6762 (1998)]. PMID- 17919055 TI - Electrodiffusion: a continuum modeling framework for biomolecular systems with realistic spatiotemporal resolution. AB - A computational framework is presented for the continuum modeling of cellular biomolecular diffusion influenced by electrostatic driving forces. This framework is developed from a combination of state-of-the-art numerical methods, geometric meshing, and computer visualization tools. In particular, a hybrid of (adaptive) finite element and boundary element methods is adopted to solve the Smoluchowski equation (SE), the Poisson equation (PE), and the Poisson-Nernst-Planck equation (PNPE) in order to describe electrodiffusion processes. The finite element method is used because of its flexibility in modeling irregular geometries and complex boundary conditions. The boundary element method is used due to the convenience of treating the singularities in the source charge distribution and its accurate solution to electrostatic problems on molecular boundaries. Nonsteady-state diffusion can be studied using this framework, with the electric field computed using the densities of charged small molecules and mobile ions in the solvent. A solution for mesh generation for biomolecular systems is supplied, which is an essential component for the finite element and boundary element computations. The uncoupled Smoluchowski equation and Poisson-Boltzmann equation are considered as special cases of the PNPE in the numerical algorithm, and therefore can be solved in this framework as well. Two types of computations are reported in the results: stationary PNPE and time-dependent SE or Nernst-Planck equations solutions. A biological application of the first type is the ionic density distribution around a fragment of DNA determined by the equilibrium PNPE. The stationary PNPE with nonzero flux is also studied for a simple model system, and leads to an observation that the interference on electrostatic field of the substrate charges strongly affects the reaction rate coefficient. The second is a time-dependent diffusion process: the consumption of the neurotransmitter acetylcholine by acetylcholinesterase, determined by the SE and a single uncoupled solution of the Poisson-Boltzmann equation. The electrostatic effects, counterion compensation, spatiotemporal distribution, and diffusion-controlled reaction kinetics are analyzed and different methods are compared. PMID- 17919057 TI - Removal forces and adhesion properties of Saccharomyces cerevisiae on glass substrates probed by optical tweezer. AB - In agroindustry, the hygiene of solid surfaces is of primary importance in order to ensure that products are safe for consumers. To improve safety, one of the major ways consists in identifying and understanding the mechanisms of microbial cell adhesion to nonporous solid surfaces or filtration membranes. In this paper we investigate the adhesion of the yeast cell Saccharomyces cerevisiae (about 5 mum in diameter) to a model solid surface, using well-defined hydrophilic glass substrates. An optical tweezer device developed by Piau [J. Non-Newtonian Fluid Mech. 144, 1 (2007)] was applied to yeast cells in contact with well characterized glass surfaces. Two planes of observation were used to obtain quantitative measurements of removal forces and to characterize the corresponding mechanisms at a micrometer length scale. The results highlight various adhesion mechanisms, depending on the ionic strength, contact time, and type of yeast. The study has allowed to show a considerable increase of adhering cells with the ionic strength and has provided a quantitative measurement of the detachment forces of cultured yeast cells. Force levels are found to grow with ionic strength and differences in mobility are highlighted. The results clearly underline that a microrheological approach is essential for analyzing the adhesion mechanisms of biological systems at the relevant local scales. PMID- 17919058 TI - Scanning electron microscopic study of air abrasion effects on human dentine. AB - AIM: Mechanical Surface Micro-Abrasion (MSM) is a technique of cavity preparation and surface treatment. By means of a precise air-powder jet it cleans and widens pits and fissures before sealing, in order to create micro-retention on tooth surface and to prepare small therapeutic cavities. It is particularly indicated in children's therapy. The aim of this study is to verify (by SEM) the existence of a relationship between working time and distance and both macroscopic and ultrastructural aspects of the treated surfaces following cavity preparation by MSM. MATERIALS AND METHODS: This experimental study was carried out in vitro using a Micro- Abrasion system on 60 human third mandibular molars. Before SEM observation the surfaces were divided in five groups, each with a different working time and distance. All specimens were observed by SEM at several magnifications. RESULTS: After a treatment of 5 sec at a distance of 2 mm a small preparation could be noticed with a circular section of 0.5 mm of diameter. With a working time of 15 sec, and a working distance of 2 mm, a cavity preparation on dentinal tissue was obtained. With a working distance of 15 mm, even for a relatively long time of treatment, such as 30 sec, no preparation was noticed but only a sandblasted surface of a circular section with a diameter of 3.5 mm. With different time of application the authors noticed different microscopic aspects. CONCLUSION: The authors realised that the macroscopic size and shape of cavities is connected to working distance, while working time is important to determine the depth of preparation and ultrastructural aspect. SEM analysis of dentin surface shows how different parameters determine macroscopic and ultrastructural aspects. It can help to standardise a protocol to follow according to the desired treatment. PMID- 17919059 TI - Comparative evaluation of different techniques of surface preparation for occlusal sealing. AB - AIM: Aim of this experiment is the efficacy evaluation of three different types of surface preparation techniques by microleakage assessment of two different sealing materials. MATERIALS AND METHODS: Sixty teeth were used for the experiment: half were treated with Admira seal (Voco) and the remaining half with Delton FS+ (Dentsply). Each group was then divided into three subgroups, which underwent three different preparation techniques before sealant placement: mechanical brushing, air abrasion and Intensive bur FG 40D4, respectively. After immersion of the specimens in methylene blue solution, the degree of microleakage was assessed by mean of optic microscope. RESULTS: Results show no microleakage in samples treated with air abrasion, followed by the samples treated with mechanical brushing. The teeth prepared by bur treatment before sealant application show a significant number and degree of marginal infiltration. CONCLUSION: It can be concluded that air abrasion technique is able to guarantee a reliable seal of occlusal surfaces. PMID- 17919060 TI - Dental caries among 12-year-old children in northern Poland between 1987 and 2003. AB - AIM: In 1987, 1995 and 2003 dental surveys were carried out in the Gdansk region of Poland investigating the prevalence of dental caries in 12-year-old school children. MATERIALS AND METHODS: Sampling and examination were performed according to WHO standards. A sample of 180 children in each year were selected from different living environments: a large city, a small town and a rural area. RESULTS: The caries prevalence (ratio of caries-free children) and caries experience (mean DMFT scores) changed significantly in some groups in the period from 1987-2003. However, there was no change in caries experience in children living in a large city. The average DMFT score of all 12-year-olds dropped from 4.07 in 1987 to 3.20 in 2003. CONCLUSION: The remarkable improvement of dental health may be explained by the oral health education and increased availability of fluoridated toothpaste. PMID- 17919061 TI - Epidemiology and aetiology of malocclusion among Syrian paediatric patients. AB - AIM: The knowledge of the distribution and causes of malocclusion in early childhood can prevent such malocclusions and their consequences and possibly to reduce complex orthodontic treatment. METHODS: All children (n = 234; 116 male, 118 female, age 2-16) who presented with occlusal or functional problems at the Department of Paediatric Dentistry at Damascus University (1995-1999) were classified according to the type and causes of malocclusion and the subsequent orthodontic treatment. RESULTS: 57.3% of the malocclusions affected the anterior posterior plane, 12.4% the vertical, and 35.9% the transverse. Most of the malocclusions had causes which could be modified by preventive or interceptive efforts: thumb sucking (5.12%), infantile swallowing (3.84%), mouth breathing (0.85%), tongue thrusting (1.7%), delayed exfoliation of a primary tooth (3.41%), premature loss of a primary tooth (9.82%), delayed eruption of a permanent tooth (2.13%), or an impacted tooth (6.41%). The subsequent early orthodontic treatment consisted of space maintainers (26%), removable (40.9%), functional (11.9%), or fixed appliances (21.2%). CONCLUSION: These data stress the importance of early orthodontic diagnosis in paediatric practice. Malocclusion in this Syrian sample could be treated to a great extent with preventive or interceptive orthodontics, which required only simple appliances and techniques. PMID- 17919062 TI - Validation of the Risk Of Malocclusion Assessment (ROMA) index. AB - AIM: The aim of this study was to validate the ROMA (Risk Of Malocclusion Assessment) Index, verifying both its reproducibility and its ability to determine treatment priority. MATERIALS AND METHODS: The levels of treatment priority assessed for 75 children examined at the Gemelli General Hospital Orthodontic Service (Rome) ranged from grade 3 to grade 5 of the index. Multivariate analysis was conducted to verify the influence of covariates on the dependent variable. Moreover, the DHC of the IOTN was assessed for all patients, and a bivariate analysis was carried out to investigate its relationship with the ROMA Index. Intra- and inter- examiner reliability was calculated on a sample of 20 children, and the reproducibility of the index was evaluated using the Kappa statistic as a measure of concordance. RESULTS: It arose that there is an inverse relation between risk grades and the time interval from the first examination and start of treatment. According to the index, patients at extreme risk require immediate treatment, whereas priority is lower in case of great or moderate risk. In statistical terms, the study revealed a significant correlation between the ROMA Index and the DHC of the IOTN. With regard to intra- and inter-examiner reliability, the data collected by the two examiners showed a high level of agreement. CONCLUSION: The ROMA index appears quick to apply, reproducible, reliable and sensitive as a mean for carefully identifying different dento skeletal problems. Furthermore, it is able to determine the priority of every risk grade and the corresponding timing of treatment. PMID- 17919063 TI - Neuromuscular diagnosis in orthodontics: effects of TENS on maxillo-mandibular relationship. AB - AIM: This study was conducted in order to assess the changes in the occlusal position of the mandible after Ultra Low Frequency (ULF)-TENS relaxing procedure in children with diagnosed functional mandibular lateral deviation. METHODS: This study was performed on 20 children, aged between 8 and 12 years, with a mean age of 10 years (SD 0.79) with functional mandibular lateral deviation, diagnosed by clinical and cephalometric evaluation, referred to the Dental Clinic for paediatric dental care. Diagnostic neuromuscular registrations were made for all children, and their casts mounted on articulator Galetti at the myocentric position and compared to casts provided of a wax bite registration in intercuspal position. RESULTS: The alignment of the midline after TENS was not punctual (p >0.05). Compared with the existing intercuspal position, neuromuscular registration showed improvement in 10 (50%) patients, 6 patients (30%) showed no changes, while worsening of the tooth-midline discrepancy was assessed in the remaining 4 (20%). The molar relationship did not follow the same trend of the midline because of the three-dimensional changes in the maxillo-mandibular relationship induced by TENS. After TENS there was a significant correlation between midline and right side deviation (r >0.65), there was no correlation between midline and the left side (r <0.65). Furthermore, right molar movement showed no correlation with the contralateral molar (r <0.65). The posterior areas of the arch were moving in a very unpredictable way, resulting in the diagnosis and prognosis of mandibular lateral deviation as absolutely individual and unpredictable. Conclusion This study suggests that TENS recorded occlusion is an interesting diagnostic approach in orthodontics since it allows visualising the trends of the neuromuscular system. PMID- 17919064 TI - Removal of fibrous epulis with Er,Cr:YSGG laser: case report. AB - BACKGROUND: Epulis is a benign tumour located in the area of the alveolar bone, periodontal ligament and marginal gingiva. A clinical case of Epulis, treated using an Er,Cr:YSGG laser in our Paediatric Dentistry division of the PTV Hospital, University of Rome Tor Vergata, is described. CASE REPORT: A pink, sessile, broad-based lesion, elastic in consistency, was detected on the maxillary vestibular gum above the lateral right incisor. The lesion was removed with Er,Cr:YSGG laser (2780 nm) without anaesthetic infiltration, power ranged from 1,5 to 2,0 Watts at 20 Hz repetition rate under 20%-15% air-water spray. The histopathological examination confirmed the diagnosis of fibrous epulis. The immediate postoperative course was excellent, with no pain or need for anti inflammatory or analgesic drugs. Wound healing was good after 1 week, and was completed after 1 month. The patient was followed up for 3-6 months, and checked again after 1 and 2 years to assess possible relapse. The Er,Cr:YSGG laser has several treatment advantages, fundamental in Paediatric Dentistry: it requires only topical anaesthesia, it has a high clinical safety, there is a short treatment time, no surgical sutures are required; no complications were encountered during or immediately following laser surgery, all resulting in excellent patient cooperation. PMID- 17919065 TI - National Italian Guidelines for caries prevention in 0 to 12 years-old children. AB - AIM: Oral and dental health improved tremendously over the last fifty years in Italy but still prevalence of dental caries in children remains a significant clinical problem. This report describes the National Italian Guidelines for caries prevention. METHODOLOGY: A panel of experts coordinated by the Italian Society of Paediatric Dentistry (SIOI) planned to elaborate the national Italian guidelines for caries prevention in children. The structure of the guidelines has been planned to follow the principles of modern caries treatment and management as well as science based dentistry. The main procedure was based on a hierarchic evaluation of literature. CONCLUSION: The guidelines are planned for dentist working in primary dental care, however, they are also designed to be of interest for other care professionals such as paediatricians, gynecologists, pharmacists and general medical practitioners and also for parents and/or guardians of the children. PMID- 17919066 TI - Nrf2-mediated heme oxygenase-1 induction confers adaptive survival response to tetrahydropapaveroline-induced oxidative PC12 cell death. AB - Tetrahydropapaveroline (THP), a dopaminergic isoquinoline neurotoxin, has been reported to contribute to neurodegeneration in parkinsonism. As THP bears two catechol moieties, it undergoes autooxidation or enzymatic oxidation to produce reactive oxygen species (ROS), which may contribute to the THP-induced cell death. Although ROS are cytotoxic, the initial accumulation of ROS may provoke a survival response. In this study, treatment of PC12 cells with THP increased expression of heme oxygenase-1 (HO-1) as an adaptive survival response. Furthermore, THP-induced cytotoxicity was attenuated by the HO-1 inducer (SnCl2) and exacerbated by the HO-1 inhibitor (ZnPP). To elucidate the molecular mechanisms underlying THP-mediated HO-1 expression, we examined the possible involvement of NF-E2-related factor 2 (Nrf2), which plays an important role in the transcriptional regulation of detoxifying/antioxidant genes. THP treatment elevated nuclear translocation of Nrf2 and subsequent binding to antioxidant response element (ARE). PC12 cells transfected with dominant-negative Nrf2 exhibited increased cytotoxicity and decreased HO-1 expression after THP treatment. Moreover, U0126 and LY294002, which are pharmacologic inhibitors of extracellular signal-regulated kinase1/2 and phosphoinositide 3-kinase, respectively, attenuated HO-1 expression as well as Nrf2-ARE binding activity. Taken together, these findings suggest that HO-1 induction via Nrf2 activation may confer a cellular adaptive response against THP-mediated cell death. PMID- 17919067 TI - Therapeutic applications of bilirubin and biliverdin in transplantation. AB - Bilirubin is the end product of heme catabolism by heme oxygenases. The inducible form of these enzymes is heme oxygenase-1 (HO-1), which is the rate-limiting enzyme that can degrade heme into equimolar quantities of carbon monoxide (CO), biliverdin, and free iron. Biliverdin is very rapidly converted to bilirubin by the enzyme biliverdin reductase, and free iron upregulates the expression of ferritin. HO-1 is a ubiquitous stress protein and is induced in many cell types by various stimuli. Induced HO-1 exerts antiinflammatory effects and modulates apoptosis. Expression of HO-1 in vivo suppresses the inflammatory responses in endotoxic shock, hyperoxia, acute pleurisy, and organ transplantation, as well as ischemia-reperfusion injury, and thereby provides salutary effects in these conditions. Accumulating evidence indicates that biliverdin/bilirubin can mediate the protective effects of HO-1 in many disease models, such as IRI and organ transplantation, via its antiinflammatory, antiapoptotic, antiproliferative, and antioxidant properties, as well as its effects on the immune response. This review attempts to summarize these protective roles as well as the molecular mechanisms by which biliverdin/bilirubin benefit IRI and solid-organ transplantation, including chronic rejection, and islet transplantation. PMID- 17919068 TI - Biliverdin reductase: PKC interaction at the cross-talk of MAPK and PI3K signaling pathways. AB - Biliverdin reductase (BVR) was characterized some 25 years ago as a unique dual cofactor/pH-dependent enzyme that catalyzes the reduction of biliverdin-IXa. Our knowledge of functions of BVR has increased enormously in recent years. hBVR functions in the IR/IGF-1-controlled regulation of the MAPK and PI3K cascades that are linked by the PKC enzymes. The first of the two culminates in the activation of transcription factors for oxidative stress-responsive genes, including ho-1, where BVR functions as both a bZip (basic leucine zipper) transcription factor and a kinase. The second pathway amplifies the insulin/growth-factor signal for protein/DNA synthesis and glucose transport downstream of PI3K. hBVR is a transactivator of PKC-betaII, and thus an integral component of the "activation loop" linking MAPK, PKC-betaII, and PI3K to insulin/growth-factor signaling. The emergence of biliverdin and bilirubin as a newly defined category of modulators of cell signaling and kinase activity further underscores the critical input of hBVR in the response of intracellular pathways into the external environment. Structural features of BVR and recent findings relevant to its function in cell-signaling pathways are reviewed here and are intended to complement a recent commentary on the role of BVR in linking heme metabolism and cell signaling. PMID- 17919069 TI - Suspension of bone marrow-derived undifferentiated mesenchymal stromal cells for repair of superficial digital flexor tendon in race horses. AB - It has been proven that mesenchymal stromal cells (MSCs) can differentiate into tenocytes. Attempts to repair tendon lesions have been performed, mainly using scaffold carriers in experimental settings. In this article, we describe the clinical use of undifferentiated MSCs in racehorses. Significant clinical recovery was achieved in 9 of 11 horses evaluated using ultrasound analysis and their ability to return to racing. Our results show that the suspension of a small number of undifferentiated MSCs may be sufficient to repair damaged tendons without the use of scaffold support. Ultrasound scanning showed that fibers were correctly oriented. By using undifferentiated cells, no ectopic bone deposition occurred. A sufficient number of cells was recovered for therapeutic purposes in all but 1 case. We suggest that the use of autologous MSCs is a safe therapeutic method for treating incompletely (i.e., not full-thickness) damaged tendons. PMID- 17919070 TI - In silico and in vitro characterization of mGBP4 splice variants. AB - In a systematic approach to identify interferon-gamma (IFN-gamma)-regulated host effector molecules, we found several members of the 65 kDa guanylate-binding proteins (GBPs) highly upregulated. During extensive characterization of these guanosine triphosphatases (GTPases), we identified discrepancies between the cloned and published sequences of the murine GTPase mGBP4. Two splice variants of mGBP4 could be detected. One variant led to a premature stop codon after 312 bp. The second variant resulted in a transcript with a disrupted G2 domain and was deposited as mGBP4.1 to the GenBank. Interestingly, only mGBP4, not mGBP4.1 mRNA, was highly upregulated in mice after infection with Listeria monocytogenes. PMID- 17919071 TI - In vitro investigation of the apoptotic effect of heparin on lymphoblasts by using flow cytometric DNA analysis and fluorometric caspase-3 and -8 activities. AB - The apoptotic effect of heparin on the lymphoblasts obtained from 12 newly diagnosed children with acute lymphoblastic leukemia (ALL) was investigated in vitro. The lymphoblasts were incubated with 0, 10, and 20 U/mL heparin concentrations at 0, 1, and 2 h. The percentages of the apoptotic lymphoblasts were calculated by flow cytometry (FCM), and activities of caspase-3 and -8 were simultaneously measured by fluorometric protease activity method. The apoptotic effect of heparin on the lymphoblasts was determined in 10 and 20 U/mL heparin concentrations (p < 0.005 and p < 0.001, respectively) while no apoptosis was detected in 0 U/mL heparin concentration at 0, 1, and 2 h. The apoptotic percentages of the lymphoblasts were higher at the first hour than those at 0 and 2 h in 10 and 20 U/mL heparin levels (p < 0.001). The highest apoptosis was found at first hour in 20 U/mL heparin concentration. Increased concentrations of heparin had an increasing effect on the percentages of the apoptotic lymphoblasts. Significantly higher caspase-3 and -8 activities were determined in 10 and 20 U/mL heparin concentrations than those in 0 U/mL heparin concentration at 0, 1, and 2 h (p < 0.001). There were no significant differences between the caspase-3 and -8 activities in 10 and 20 U/mL heparin concentrations at 1 and 2 h (p > 0.05), while statistically significant differences were simultaneously detected in the apoptotic rates of the lymphoblasts (p < 0.001). This may be due to that the study included the limited patients, or measurement of the caspase activities is a more sensitive method than the FCM analysis for determination of apoptosis because the activation time of the caspases takes a long time period. It was concluded that the apoptotic effect of heparin in vitro on lymphoblasts developed due to the extrinsic pathway of apoptosis via the caspase-3 and -8 activations in newly diagnosed ALL patients. PMID- 17919072 TI - Nordihydroguaiaretic acid increases endothelial nitric oxide synthase expression via the transcription factor AP-1. AB - It has been previously reported that the antioxidant compound nordihydroguaiaretic acid (NDGA) increases endothelial nitric oxide synthase (eNOS) expression in cultured bovine aortic endothelial cells. However, the exact mechanism for this effect was unresolved. Thus, the purpose of this study was to further elucidate the effect of NDGA on eNOS protein expression and enzymatic activity in fetal pulmonary arterial endothelial cells (FPAECs), and to identify the transcription factors involved in this regulation. Following overnight exposure to 0-32 microM NDGA, we observed a 2- to 2.5-fold increase in eNOS protein expression in FPAECs, with a similar increase observed in eNOS activity. For eNOS gene promoter analysis, we initially used two promoter-reporter constructs: a 1.6 kb promoter fragment and an 840 bp construct, both of which include an AP-1-specific binding site. NDGA exposure induced a significant increase in eNOS promoter activity in both constructs. However, the NDGA-mediated increase was abolished when we used either a truncated promoter construct lacking the AP-1 element or a construct in which the AP-1 binding site was mutated. AP-1 binding efficiency was also determined by electrophoretic mobility shift assay, where we observed an increase in AP-1 binding in FPAECs treated with NDGA while the binding of AP-1 was found to be decreased when a mutated AP-1 consensus sequence was used. Further, supershift analyses indicated that the AP-1 complex consisted of c-Jun and FosB. We therefore conclude that NDGA antioxidant activity regulates eNOS expression via AP-1 and that antioxidant therapy could potentially be used to increase eNOS expression in diseases, such as persistent pulmonary hypertension of the newborn, where eNOS expression and activity are known to be reduced. PMID- 17919073 TI - Cytochrome P4501A1 and microsomal epoxide hydrolase gene polymorphisms: gene environment interaction and risk of prostate cancer. AB - The role of low penetrance genes and environmental factors in the etiology of prostate cancer (PCa) is unclear. Most procarcinogens require metabolic activation by CYP4501A1, whereas microsomal epoxide hydrolase (mEH) is involved in the detoxification. In our case-control study, we assessed whether CYP1A1 and mEH susceptibility genotypes, tobacco use, and age factors contribute to PCa risk. One hundred thirty patients with PCa and 140 control subjects were analyzed by polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) method from genomic DNA samples. Binary logistic regression model was used for assessing differences in genotype prevalence and their association between patient and the control group. T/C polymorphism of CYP1A1 gene revealed significant association with the tobacco users (p < 0.005) for PCa risk. Our results demonstrated significant association with exon 3 variant genotypes of the mEH alone or in combination with tobacco users (p < 0.005), whereas in exon 4 genotypes, no association was observed. Haplotype analysis projected significant associations with very slow haplotypes of mEH gene (OR = 2.48, 95% CI = 1.41 4.38, p = 0.002). In conclusion, our study demonstrated that exon 3 of mEH and CYP1A1 T/C gene polymorphism are predisposing risk factors for susceptibility of sporadic PCa in northern India. It also suggests that a combination of smoking plays a significant role in modified PCa risk on the study population, which means that smokers carrying susceptible genotypes may be subjected to higher risk than those carrying nonsusceptible genotypes. PMID- 17919074 TI - Effects of varying the experimental design of a cognitive control paradigm on behavioral and functional imaging outcome measures. AB - Abstract A number of experimental techniques are commonly used within the field of functional neuroimaging to measure successive cognitive processes within a single trial. This study evaluated three experimental techniques to assess the comparability of behavioral and functional outcome measures in a task involving higher-level cognitive processing while controlling for the task duration. Twelve participants completed a cognitive control paradigm using the three techniques. Each trial of the task consisted of a green or red cue followed by a "Left" or "Right" probe. Green cues indicated that participants should respond in the direction of the probe. Red cues indicated participants should overcome their automatic tendency and respond in the direction opposite to the probe. The "slow" technique involved a sufficiently long trial allowing the blood oxygenation level dependent response to rise and return to baseline before the next trial. The "jitter" technique involved varying the interstimulus and intertrial intervals. The "catch" technique involved presenting some cue-only trials in the midst of cue-probe trials. Predicted brain regions were activated by all the experimental techniques combined including the middle frontal, anterior cingulate, and inferior parietal cortices. Although there were more commonalties than differences between the three experimental techniques, generally, it appeared that the slow technique was effective at detecting posterior activity; the jitter technique was effective at detecting probe-related activity; and the catch technique was effective at detecting cue-related activity, especially in prefrontal regions. Thus, experiments measuring successive cognitive processes may have differential detection power for every event in a trial. PMID- 17919075 TI - Simultanagnosia: when a rose is not red. AB - Abstract Information regarding object identity ("what") and spatial location ("where/how to") is largely segregated in visual processing. Under most circumstances, however, object identity and location are linked. We report data from a simultanagnosic patient (K.E.) with bilateral posterior parietal infarcts who was unable to "see" more than one object in an array despite relatively preserved object processing and normal preattentive processing. K.E. also demonstrated a finding that has not, to our knowledge, been reported: He was unable to report more than one attribute of a single object. For example, he was unable to name the color of the ink in which words were written despite naming the word correctly. Several experiments demonstrated, however, that perceptual attributes that he was unable to report influenced his performance. We suggest that binding of object identity and location is a limited-capacity operation that is essential for conscious awareness for which the posterior parietal lobe is crucial. PMID- 17919076 TI - Neural and behavioral evidence for affective priming from unconsciously perceived emotional facial expressions and the influence of trait anxiety. AB - Abstract Affective judgments can often be influenced by emotional information people unconsciously perceive, but the neural mechanisms responsible for these effects and how they are modulated by individual differences in sensitivity to threat are unclear. Here we studied subliminal affective priming by recording brain potentials to surprise faces preceded by 30-msec happy or fearful prime faces. Participants showed valence-consistent changes in affective ratings of surprise faces, although they reported no knowledge of prime-face expressions, nor could they discriminate between prime-face expressions in a forced-choice test. In conjunction with the priming effect on affective evaluation, larger occipital P1 potentials at 145-175 msec were found with fearful than with happy primes, and source analyses implicated the bilateral extrastriate cortex in this effect. Later brain potentials at 300-400 msec were enhanced with happy versus fearful primes, which may reflect differential attentional orienting. Personality testing for sensitivity to threat, especially social threat, was also used to evaluate individual differences potentially relevant to subliminal affective priming. Indeed, participants with high trait anxiety demonstrated stronger affective priming and greater P1 differences than did those with low trait anxiety, and these effects were driven by fearful primes. Results thus suggest that unconsciously perceived affective information influences social judgments by altering very early perceptual analyses, and that this influence is accentuated to the extent that people are oversensitive to threat. In this way, perception may be subject to a variety of influences that govern social preferences in the absence of concomitant awareness of such influences. PMID- 17919078 TI - Lexical plasticity in early bilinguals does not alter phoneme categories: I. Neurodynamical modeling. AB - Abstract Sebastian-Galles et al. [The influence of initial exposure on lexical representation: Comparing early and simultaneous bilinguals. Journal of Memory and Language, 52, 240-255, 2005] contrasted highly proficient early Spanish Catalan and Catalan-Spanish bilinguals, using Catalan materials in a lexical decision task (LDT). They constructed two types of experimental pseudowords, substituting Catalan phoneme /e/ for Catalan /epsilon/, or vice versa. Catalan dominant bilinguals showed a performance asymmetry across experimental conditions, making more mistakes for /epsilon/-->/e/ changes, than for /e/- >/epsilon/ ones. This was considered evidence of a developed acceptance of mispronounced Catalan /epsilon/-words, caused by exposure to a bilingual environment where mispronunciations by Spanish-dominant bilinguals using their /e/-category abound. Although this indicated modified or added lexical representations, an open issue is whether such lexical information also modifies phoneme categories. We address this using a biophysically realistic neurodynamic model, describing neural activity at the synaptic and spiking levels. We construct a network of pools of neurons, representing phonemic and lexical processing. Carefully analyzing the dependency of network dynamics on connection strengths, by first exploring parameter space under steady-state assumptions (mean-field scans), then running spiking simulations, we investigate the neural substrate role in a representative LDT. We also simulate a phoneme discrimination task to address whether lexical changes affect the phonemic level. We find that the same network configuration which displays asymmetry in the LDT shows equal performance discriminating the two modeled phonemes. Thus, we predicted that the Catalan-dominant bilinguals do not alter their phoneme categories, although showing signs of having stored a new word variation in the lexicon. To explore this prediction, a syllable discrimination task involving the /e/-/epsilon/ contrast was set up, using Catalan-dominants displaying performance asymmetry in a repetition of the original LDT. Discrimination task results support the prediction, showing that these subjects discriminate both categories equally well. We conclude that subjects often exposed to dialectal word variations can store these in their lexicons, without altering their phoneme representations. PMID- 17919079 TI - Interactive processing of timbre dimensions: an exploration with event-related potentials. AB - Abstract Timbre characterizes the identity of a sound source. On psychoacoustic grounds, it has been described as a multidimensional perceptual attribute of complex sounds. Using Garner's interference paradigm, we found in a previous behavioral study that three timbral dimensions exhibited interactive processing. These timbral dimensions acoustically corresponded to attack time, spectral centroid, and spectrum fine structure. Here, using event-related potentials (ERPs), we sought neurophysiological correlates of the interactive processing of these dimensions of timbre. ERPs allowed us to dissociate several levels of interaction, at both early perceptual and late stimulus identification stages of processing. The cost of filtering out an irrelevant timbral dimension was accompanied by a late negative-going activity, whereas congruency effects between timbre dimensions were associated with interactions in both early sensory and late processing stages. ERPs also helped to determine the similarities and differences in the interactions displayed by the different pairs of timbre dimensions, revealing in particular variations in the latencies at which temporal and spectral timbre dimensions can interfere with the processing of another spectral timbre dimension. PMID- 17919080 TI - The role of the right cerebral hemisphere in processing novel metaphoric expressions: a transcranial magnetic stimulation study. AB - Abstract Previous research suggests that the right hemisphere (RH) may contribute uniquely to the processing of metaphoric language. However, causal relationships between local brain activity in the RH and metaphors comprehension were never established. In addition, most studies have focused on familiar metaphoric expressions which might be processed similarly to any conventional word combination. The present study was designed to overcome these two problems by employing repetitive transcranial magnetic stimulation (rTMS) to examine the role of the RH in processing novel metaphoric expressions taken from poetry. Right handed participants were presented with four types of word pairs, literal, conventional metaphoric and novel metaphoric expressions, and unrelated word pairs, and were asked to perform a semantic judgment task. rTMS of the right posterior superior temporal sulcus disrupted processing of novel but not conventional metaphors, whereas rTMS over the left inferior frontal gyrus selectively impaired processing of literal word pairs and conventional but not novel metaphors (Experiment 1). In a further experiment, we showed that these effects were due to right-left asymmetries rather than posterior-anterior differences (Experiment 2). This is the first demonstration of TMS-induced impairment in processing novel metaphoric expressions, and as such, confirms the specialization of the RH in the activation of a broader range of related meanings than the left hemisphere, including novel, nonsalient meanings. The findings thus suggest that the RH may be critically involved in at least one important component of novel metaphor comprehension, the integration of the individual meanings of two seemingly unrelated concepts into a meaningful metaphoric expression. PMID- 17919081 TI - To ignore or explore: top-down modulation of novelty processing. AB - Abstract Attending to novelty is a critical element of human behavior and learning. Novel events can serve as task-irrelevant distracters or as potential sources of engagement by interesting or important aspects of one's environment. An optimally functioning brain should have the capacity to respond differentially to novel events depending on the circumstances in which they occur. In the present study, a subject-controlled variant of the visual novelty oddball paradigm was employed under two different conditions in which novel stimuli were characterized either as distracters from a main task or as potentially meaningful "invitations" to explore the environment. Differences in context, derived from varying the emphasis of task instructions, strongly modulated both the behavioral and electrophysiological response to novelty. This modulation was not observed for processing earlier than the P3 component. Subjects who encountered novel events that served as distracters limited the amount of attention and processing resources they appropriated. Remarkably, under this condition, there were no differences in overall P3 amplitude, late positive slow-wave activity, or viewing duration between rare novel and frequent standard events. In contrast, subjects who encountered novel events as potential opportunities to explore augmented the attention and processing resources directed toward these events (as reflected by a larger P3 amplitude, late positive slow-wave activity, and longer viewing durations). Our results suggest that the processing of novelty within the visual modality involves several stages, including: (1) the relatively automatic detection of unfamiliar, novel stimuli (indexed by the N2); (2) the voluntary allocation of resources determined by the broader context in which a novel event occurs (indexed by the P3); and (3) the sustained processing of novelty (indexed by late positive slow-wave activity). This study provides evidence of the brain's ability to generate differential responses to novel events according to the circumstances under which they are encountered. It also points to a greater degree of top-down modulation of the processing of novelty than has been previously emphasized. We suggest that less commonly studied variables, such as subject control, may provide additional insight into the different ways in which novelty is processed. PMID- 17919082 TI - Neuroimaging studies of mental rotation: a meta-analysis and review. AB - Abstract Mental rotation is a hypothesized imagery process that has inspired controversy regarding the substrate of human spatial reasoning. Two central questions about mental rotation remain: Does mental rotation depend on analog spatial representations, and does mental rotation depend on motor simulation? A review and meta-analysis of neuroimaging studies help answer these questions. Mental rotation is accompanied by increased activity in the intraparietal sulcus and adjacent regions. These areas contain spatially mapped representations, and activity in these areas is modulated by parametric manipulations of mental rotation tasks, supporting the view that mental rotation depends on analog representations. Mental rotation also is accompanied by activity in the medial superior precentral cortex, particularly under conditions that favor motor simulation, supporting the view that mental rotation depends on motor simulation in some situations. The relationship between mental rotation and motor simulation can be understood in terms of how these two processes update spatial reference frames. PMID- 17919084 TI - Linking cortical spike pattern codes to auditory perception. AB - Abstract Neurometric analysis has proven to be a powerful tool for studying links between neural activity and perception, especially in visual and somatosensory cortices, but conventional neurometrics are based on a simplistic rate-coding hypothesis that is clearly at odds with the rich and complex temporal spiking patterns evoked by many natural stimuli. In this study, we investigated the possible relationships between temporal spike pattern codes in the primary auditory cortex (A1) and the perceptual detection of subtle changes in the temporal structure of a natural sound. Using a two-alternative forced-choice oddity task, we measured the ability of human listeners to detect local time reversals in a marmoset twitter call. We also recorded responses of neurons in A1 of anesthetized and awake ferrets to these stimuli, and analyzed these responses using a novel neurometric approach that is sensitive to temporal discharge patterns. We found that although spike count-based neurometrics were inadequate to account for behavioral performance on this auditory task, neurometrics based on the temporal discharge patterns of populations of A1 units closely matched the psychometric performance curve, but only if the spiking patterns were resolved at temporal resolutions of 20 msec or better. These results demonstrate that neurometric discrimination curves can be calculated for temporal spiking patterns, and they suggest that such an extension of previous spike count-based approaches is likely to be essential for understanding the neural correlates of the perception of stimuli with a complex temporal structure. PMID- 17919083 TI - Bilingual and monolingual brains compared: a functional magnetic resonance imaging investigation of syntactic processing and a possible "neural signature" of bilingualism. AB - Abstract Does the brain of a bilingual process language differently from that of a monolingual? We compared how bilinguals and monolinguals recruit classic language brain areas in response to a language task and asked whether there is a "neural signature" of bilingualism. Highly proficient and early-exposed adult Spanish-English bilinguals and English monolinguals participated. During functional magnetic resonance imaging (fMRI), participants completed a syntactic "sentence judgment task" [Caplan, D., Alpert, N., & Waters, G. Effects of syntactic structure and propositional number on patterns of regional cerebral blood flow. Journal of Cognitive Neuroscience, 10, 541-552, 1998]. The sentences exploited differences between Spanish and English linguistic properties, allowing us to explore similarities and differences in behavioral and neural responses between bilinguals and monolinguals, and between a bilingual's two languages. If bilinguals' neural processing differs across their two languages, then differential behavioral and neural patterns should be observed in Spanish and English. Results show that behaviorally, in English, bilinguals and monolinguals had the same speed and accuracy, yet, as predicted from the Spanish-English structural differences, bilinguals had a different pattern of performance in Spanish. fMRI analyses revealed that both monolinguals (in one language) and bilinguals (in each language) showed predicted increases in activation in classic language areas (e.g., left inferior frontal cortex, LIFC), with any neural differences between the bilingual's two languages being principled and predictable based on the morphosyntactic differences between Spanish and English. However, an important difference was that bilinguals had a significantly greater increase in the blood oxygenation level-dependent signal in the LIFC (BA 45) when processing English than the English monolinguals. The results provide insight into the decades-old question about the degree of separation of bilinguals' dual language representation. The differential activation for bilinguals and monolinguals opens the question as to whether there may possibly be a "neural signature" of bilingualism. Differential activation may further provide a fascinating window into the language processing potential not recruited in monolingual brains and reveal the biological extent of the neural architecture underlying all human language. PMID- 17919085 TI - Inhibition of return arises from inhibition of response processes: an analysis of oscillatory beta activity. AB - Abstract In the orienting of attention paradigm, inhibition of return (IOR) refers to slowed responses to targets presented at the same location as a preceding stimulus. No consensus has yet been reached regarding the stages of information processing underlying the inhibition. We report the results of an electro-encephalogram experiment designed to examine the involvement of response inhibition in IOR. Using a cue-target design and a target-target design, we addressed the role of response inhibition in a location discrimination task. Event-related changes in beta power were measured because oscillatory beta activity has been shown to be related to motor activity. Bilaterally located sources in the primary motor cortex showed event-related beta desynchronization (ERD) both at cue and target presentation and a rebound to event-related beta synchronization (ERS) after movement execution. In both designs, IOR arose from an enhancement of beta synchrony. IOR was related to an increase of beta ERS in the target-target design and to a decrease of beta ERD in the cue-target design. These results suggest an important role of response inhibition in IOR. PMID- 17919086 TI - A role for JAK2 mutations in myeloproliferative diseases. AB - Myeloproliferative disorders (MPDs) are characterized by a clonal expansion of myeloid cells. Over the past two years, the identification of the JAK2V617F mutation in most cases of polycythemia vera (PV) as well as approximately 50% of patients with essential thrombocythemia (ET) and idiopathic myelofibrosis (IMF) has greatly advanced our understanding of MPDs. The JAK2V617F mutation alters the JAK2 tyrosine kinase to confer constitutive activation and affect downstream signaling pathways. Data from mouse models demonstrate that the mutation is sufficient for development of PV, but additional work is needed to better understand how this allele functions in ET and IMF. Regardless of the various pathologies, the JAK2V617F discovery highlights the importance of JAK-STAT signaling in myeloid differentiation and focuses effort on developing a clinically relevant JAK2 inhibitor. PMID- 17919087 TI - PKC isozymes in chronic cardiac disease: possible therapeutic targets? AB - Cardiovascular disease is the leading cause of death in the United States. Therefore, identifying therapeutic targets is a major focus of current research. Protein kinase C (PKC), a family of serine/threonine kinases, has been identified as playing a role in many of the pathologies of heart disease. However, the lack of specific PKC regulators and the ubiquitous expression and normal physiological functions of the 11 PKC isozymes has made drug development a challenge. Here we discuss the validity of therapeutically targeting PKC, an intracellular signaling enzyme. We describe PKC structure, function, and distribution in the healthy and diseased heart, as well as the development of rationally designed isozyme selective regulators of PKC functions. The review focuses on the roles of specific PKC isozymes in atherosclerosis, fibrosis, and cardiac hypertrophy, and examines principles of pharmacology as they pertain to regulators of signaling cascades associated with these diseases. PMID- 17919088 TI - Review: immunologic response to protease inhibitor-based highly active antiretroviral therapy: a review. AB - A substantial body of evidence indicates that CD4 cell count is an important independent prognostic indicator for progression of HIV disease. Consequently, in addition to plasma HIV RNA levels, CD4 cell count change is considered to be a key surrogate marker for disease progression in clinical practice and in clinical studies. Given the relationship between changes in CD4 count and disease progression, it is notable that protease inhibitor (PI)-based highly active antiretroviral therapy (HAART) can rapidly increase CD4 cell count early in treatment in both therapy-naive and -experienced patients, and can sustain clinically relevant levels beyond 24 weeks. A number of trials with a follow-up of more than 3 years allow us to conclude that the gains in CD4 counts are maintained in a durable manner. This review evaluated randomized studies of PI based and PI-boosted HAART (published between January 1996 and February 2006) to determine the effect of PI-based therapy on CD4 cell count. Only studies that assessed CD4 response in the overall patient population were included. Four mechanisms have been proposed to account for the rapid increase in CD4 cell count that occurs with HAART: CD4 cell redistribution from lymphatic tissues, increased CD4 cell production, reduction of apoptotic CD4 cells and the recovery of hematopoietic activity in bone marrow. Further research is required to clarify the relative importance of these mechanisms and ways in which they might be enhanced. PMID- 17919089 TI - Outcomes of blood and oral fluid rapid HIV testing: a literature review, 2000 2006. AB - Rapid HIV antibody testing is a new technology whose implementation promises to facilitate the early detection of persons with HIV/AIDS. The goal of this study was to review and summarize the literature from 2000-2006 regarding four outcomes of rapid (including both blood and oral fluid) HIV testing: rates of client acceptance; rates of clients' receiving their test results; rates of entry into medical care for those found to be HIV positive; and the efficacy of prevention counseling after testing. A total of 116 studies in peer-reviewed journals were screened. Twenty-six met the screening criteria (published in peer-reviewed journals and focused on at least one of the outcomes of interest) and were selected for review. Considerable variation was found in client acceptance rates with the highest rates among pregnant women in labor and delivery units and the lowest rates in needle exchange and bath-house settings. The evidence shows that most persons tested with a rapid test receive their test result. Three studies on entry into medical care among those who were newly identified HIV positive found rates of 47%, 82%, and 97% of clients adhering to their first medical appointment. No long-term medical follow-up studies were found. Only one study examined the efficacy of prevention counseling after rapid testing and found no statistically significant differences in the number of sexually transmitted diseases (STDs) conventional versus rapid HIV testers contracted following testing. PMID- 17919090 TI - Maternal HIV-1 DNA load and mother-to-child transmission. AB - While many factors contribute to mother-to-child transmission (MTCT) of HIV-1, maternal plasma HIV-1 RNA viral load (RNA-VL) has been consistently found as the main risk factor, including when antiretroviral prophylaxis was used to prevent MTCT. However the predictive value of RNA-VL is poor. A recent study of HIV-1 positive pregnant women who did not receive antiretroviral prophylaxis reported an association between HIV-1 DNA viral load (DNA-VL) and MTCT that was stronger than the association between RNA-VL and MTCT. We sought to determine if HIV-1 DNA VL was independently associated with MTCT of HIV in a population of women who received zidovudine prophylaxis during pregnancy and whose infants received zidovudine after birth. Patients were 33 non-breastfeeding transmitting (TR) and 33 nontransmitting mothers (NTR) from Perinatal HIV Prevention Trial (PHPT-1), a multicenter clinical trial conducted in Thailand comparing zidovudine prophylaxis durations to prevent MTCT. TR and NTR mothers were matched according to baseline RNA-VL. Maternal peripheral blood mononuclear cell (PBMC)-associated HIV-1 DNA was extracted from whole blood, and DNA-VL was established by quantitative real time polymerase chain reaction. We found that TR had a significantly higher cell associated HIV-1 DNA viral load than did NTR. Median TR DNA-VL was 2.54 log(10) copies per microgram PBMC DNA, while it was 2.28 log(10) copies per microgram PBMC DNA in NTR (Wilcoxon p = 0.02). In summary, HIV-1 DNA viral load was associated with MTCT in a population of women who received antiretroviral prophylaxis during pregnancy, independently from RNA viral load. PMID- 17919091 TI - Prevalence of drug-resistant and nonsubtype B HIV strains in antiretroviral naive, HIV-infected individuals in New York State. AB - The duration of HIV infection is usually unknown for most patients entering into HIV care. Data on the frequency at which resistance mutations are detected in these patients are needed to support practical guidance on the use of resistance testing in this clinical situation. Furthermore, little is known about HIV subtype diversity in much of the United States. Therefore, we analyzed the prevalence of drug resistance mutations and nonsubtype B strains of HIV among antiretroviral-naive individuals presenting for HIV care in New York State between September 2000 and January 2004. Sequences were obtained using a commercial HIV genotyping assay. Seventeen of 151 subjects (11.3%; 95% confidence interval 7.2%-17.3%) had at least one drug-resistance mutation, including 5 subjects with fewer than 200 CD4(+) T cells, indicative of advanced infection. Nucleoside reverse transcriptase inhibitor, non-nucleoside reverse transcriptase inhibitor, and protease inhibitor resistance mutations were detected in 6.6%, 5.3%, and 0.7% of subjects, respectively. Subjects from New York City-based clinics were less likely to have resistant virus than subjects from clinics elsewhere in New York State. Nonsubtype B strains of HIV were detected in 9 (6.0%) individuals and were associated with heterosexual contact. Two nonsubtype B strains from this cohort also carried drug-resistance mutations. These data indicate that drug-resistant virus is frequently detected in antiretroviral-naive individuals entering HIV care in New York State. Furthermore, a diverse set of nonsubtype B strains were identified and evidence suggests that nonsubtype B strains, including those carrying drug-resistance mutations, are being transmitted in New York State. PMID- 17919092 TI - MDR1 and CYP3A4 polymorphisms are associated with HIV seropositivity in Israeli patients but do not influence the course of HIV disease. AB - Recent studies have examined the relationship between polymorphic alleles of the MDR1 gene and the course of HIV. Such polymorphisms may alter the metabolism of antiretroviral medications or influence susceptibility to HIV infectivity. We therefore studied a polymorphism in MDR1 (C3435T), and the CYP3A4*1B variant allele, the latter of which has not been previously studied in HIV. Ninety-six patients of either Ethiopian (57) or Caucasian (39) ethnicity and 276 controls were studied including serial CD4 counts, clinical course, and AIDS-defining illnesses. For both ethnic groups, the C allele of MDR1 C3435T was highly associated with being infected with HIV (p < 0.0001) compared to controls, but genotype did not influence change in CD4 counts over time in the patients, whether or not they were treated with antiretrovirals. CYP3A4*1B was also significantly associated with being infected with HIV (p < 0.0001) both in heterozygotes and in homozygotes for the polymorphism, but only for Ethiopians (p < 0.023 compared to Caucasians, p = 0.44). CYP3A4*1B did not influence CD4 count or AIDS defining illnesses. We conclude that in Israeli patients, polymorphisms in drug metabolism and disposition genes may influence infectivity of HIV but do not influence the course of the disease. Larger studies are needed to confirm these findings. PMID- 17919093 TI - The effects of smoking abstinence on symptom burden and quality of life among persons living with HIV/AIDS. AB - Persons living with HIV/AIDS who are current smokers are more likely to develop disease-related adverse health outcomes compared to nonsmokers with HIV/AIDS. However, the impact of smoking cessation on health outcomes such as symptom status and health-related quality of life (HRQOL) has not yet been assessed within this population. This study examined the effects of changes in smoking status on HIV-related symptom burden and health-related quality of life outcomes in a multiethnic, low-income population of persons living with HIV/AIDS. Patients (n = 95) from a large, inner city HIV/AIDS clinic were enrolled in a smoking cessation trial providing nicotine replacement therapy, counseling, and self-help written materials. Biochemically verified smoking status, length of smoking abstinence, HIV-related symptom burden, and HRQOL were assessed approximately 3 months posttrial enrollment. A series of multiple linear regression models was performed to assess the associations between the smoking status variables and the health outcomes at follow-up while controlling for baseline levels. Length of smoking abstinence was significantly associated (p = 0.02) with HIV-related symptom burden. Specifically, increasing number of consecutive days of smoking abstinence during the 3-month follow-up period was associated with lower levels of HIV-related symptom burden at the time of follow-up. However, 24-hour smoking prevalence was not significantly (p > 0.05) associated with changes in either HIV related symptom burden or HRQOL. These findings suggest that smoking cessation can significantly improve symptom burden for individuals living with HIV/AIDS. Moreover, these benefits are observable as early as 3 months after quitting and are positively correlated with the length of abstinence. PMID- 17919094 TI - Social support, coping, and medication adherence among HIV-positive women with depression living in rural areas of the southeastern United States. AB - This study examined the relationships among sociodemographic factors, social support, coping, and adherence to antiretroviral therapy (ART) among HIV-positive women with depression. The analyses reported here were limited to the 224 women receiving ART of 280 women recruited from community-based HIV/AIDS organizations serving rural areas of three states in the southeastern United States. Two indicators of medication adherence were measured; self-report of missed medications and reasons for missed medications in the past month. Descriptive statistics, correlation, and regression analyses were performed to systematically identify sociodemographic, coping, and social support variables that predicted medication adherence. In regression analysis, three variables were determined to be significant predictors accounting for approximately 30% of the variability in the self-report of reasons for missed medications. Coping focused on managing HIV disease was negatively associated, while coping focused on avoidance/denial and number of children were positively associated with reasons for missed medications. Coping by spiritual activities and focusing on the present mediated the effect of social support on self-reported missed medications. The relationship of predictor variables to self-report of missed medications was assessed using t test statistics and logistic regression analysis to determine the odds of self-reported medication adherence. Satisfaction with social support (p = 0.04), and coping focused on managing HIV disease (p = 0.002) were the best positive predictors, whereas number of children (p = 0.02) was the lone significant negative predictor of medication adherence. The study findings have implications for designing, implementing, and testing interventions based on social support and coping theories for achieving better adherence to HIV medications. PMID- 17919095 TI - Predictors of AIDS-related morbidity and mortality in a southern U.S. Cohort. AB - Advances in the treatment of HIV and associated opportunistic infections (OIs) have led to dramatic reductions in HIV-related morbidity and mortality in the United States, but not all patients have benefited equally. A longitudinal analysis of the Coping with HIV/AIDS in the Southeast (CHASE) cohort evaluated sociodemographic, psychosocial, and clinical factors associated with HIV-related events (incident category C OI or AIDS-related death) among southern HIV-infected patients engaged in clinical care. Participants were followed for a median of 30 months (interquartile range, 17-34 months) after study enrollment (enrollment period December 2001 to April 2002). Ten percent of study participants (50/489) experienced an HIV-related event (incident category C OI and/or AIDS-related deaths) during study follow-up. The rate of HIV-related events was 4.8 per 100 patient-years of observation, and the rate of AIDS-related death was 1.5 per 100 patient-years of observation. In unadjusted survival analyses, younger age, lacking private health insurance, psychosocial trauma, depressive symptoms, lower baseline CD4 count, and less time on antiretroviral therapy during follow-up were associated with HIV-related events. In Cox proportional hazards analysis adjusting for covariates, patients who had suffered more psychosocial trauma (hazard ratio [HR] = 1.97, p = 0.04), who had lower baseline CD4 counts (HR = 0.48 per 100 cells/mm(3), p < 0.01), and who spent less time on antiretroviral therapy during follow-up (HR = 0.47, p = 0.02) were more likely to experience an HIV-related event. PMID- 17919096 TI - Retention challenges for a community-based HIV primary care clinic and implications for intervention. AB - The present study sought to elucidate factors involved in loss to follow-up (LTF) among HIV-infected patients who had been receiving medical care at Fenway Community Health (FCH) located in Boston, Massachusetts. FCH provided care to 1143 HIV-infected patients in 2005, predominantly Caucasian men who have sex with men (MSM). Two approaches were used to address the research question. First, 495 patients were identified that had been LTF from 2001-2005. One hundred seventy nine eligible patients completed a questionnaire to determine reasons for discontinuing care, representing a 51% response rate. Second, a cohort study was performed using the medical record data of 896 HIV-infected patients who were receiving medical care in the year 2000. Patients' utilization of primary medical care was followed until January 1, 2005 and predictors of LTF were examined using Cox proportional hazards regression modeling. Survey respondents reported that the greatest perceived barriers to care at FCH were personal/cultural, structural, and financial in nature. Twenty-two percent reported sporadic care elsewhere with gaps in care of 6 months or more, and 8% reported no regular provider for HIV. Significant predictors of LTF from regression analysis included: minority race/ethnicity, use of safety-net insurance, appointment nonadherence and no medical social work visits. To improve engagement and retention in care, organizations may use patient surveys for organizational self assessment to effect operational changes that minimize barriers to care. A risk assessment tool based on evidence-based methods can be implemented to identify high-risk patients for innovative outreach interventions. The primary study limitation is the underrepresentation of minority and traditionally underserved populations. PMID- 17919098 TI - Possible hepatitis C vaccine. PMID- 17919097 TI - FDA approves maraviroc tablets. PMID- 17919099 TI - Amino acid conservation in the gp41 transmembrane protein and natural polymorphisms associated with enfuvirtide resistance across HIV-1 variants. AB - Information about gp41 variability across distinct HIV-1 subtypes is scarce, and yet such knowledge would be desirable for designing new drugs targeting this viral protein. Conserved gp41 residues in viruses derived from 79 individuals infected with distinct HIV-1 subtypes (29 A, 25 B, 8 C, 3 D, 4 F, 4 G, 2 H, 1 J, 1 U, and 2 CRF06_cpx) and naive for entry inhibitors were examined. Conservation of gp41 was also examined in 908, 56, and 3 HIV-1 group M, O, and N sequences, respectively, available at the Los Alamos HIV Sequence Database. Among the 345 residues in the full gp41 protein, 36% showed up to 90% conservation in all 987 group M sequences, as did 40% of 56 group O sequences and 49% of 3 group N sequences. The HR1 region (residues 29-82) showed a higher proportion of highly conserved residues than did the HR2 region (residues 116-161) in all groups (65 vs. 34% in group M, 57 vs. 46% in group O, and 80 vs. 52% in group N). Some secondary resistance mutations to enfuvirtide were found as natural polymorphisms (A30V and Q56K/R in group M, Q56R and S138A in group O, and S138A in group N). In fact, A30V was a signature change in clade G and CRF06_cpx, whereas Q56K/R was a signature change for clades A and J, as well as for CRF04_cpx, CRF09_cpx, CRF11_cpx, and CRF13_cpx. The relative conservation of amino acids in gp41 across HIV-1 variants indirectly highlights the critical role of this protein for HIV infectivity and makes it feasible to design new entry inhibitors with activity against diverse HIV-1 variants. PMID- 17919100 TI - Hospital-based surveillance for HTLV-1/2 infections in Spain. AB - In Western countries, HTLV-1 infection is recognized mainly among foreigners coming from endemic areas. In contrast, HTLV-2 is found predominantly in native intravenous drug users (IDUs). Spain has experienced a large wave of immigration, which could have influenced the current prevalence and distribution of HTLV-1 and HTLV-2 infection. A 1-day cross-sectional survey was carried out in May 2005 in 13 hospitals distributed across Spain. A total of 2873 outpatient subjects were screened for HTLV-1/2 antibodies. Although the majority of the study population consisted of native Spaniards, 206 (7.2%) were immigrants. Two cases of HTLV-1 and one of HTLV-2 infection were found (overall prevalence, 0.1%). The two individuals with HTLV-1 were immigrants from endemic areas and the single case of HTLV-2 infection was a former Spaniard IDU coinfected with HIV-1. In summary, the current prevalence of HTLV-1/2 infection in Spain is low, with no evidence of spread beyond the classical risk groups. However, a rapidly growing population of immigrants from HTLV-1-endemic areas in Spain could modify this pattern and periodic surveillance studies including both natives and immigrants are warranted. PMID- 17919101 TI - Dynamics of drug-resistant HIV-1 in plasma and peripheral blood cells in patients during and after enfuvirtide therapy. AB - Genetic analysis of HIV-1 gp41 in plasma and peripheral blood mononuclear cell (PBMC) compartments was performed longitudinally in 10 HIV-infected patients treated with enfuvirtide. The appearance of enfuvirtide resistance mutations in PBMCs (DNA) generally occurred after (from 23 to 157 weeks) being recognizable in plasma (RNA). The disappearance of drug-resistant HIV-1 mutations to enfuvirtide in seven patients who discontinued the drug was directly dependent of the exposure time failing enfuvirtide, being associated mainly with the selection of secondary/compensatory mutations recognizable in proviral DNA. PMID- 17919102 TI - Comparative longitudinal studies of HERV-K and HIV-1 RNA titers in HIV-1-infected patients receiving successful versus unsuccessful highly active antiretroviral therapy. AB - The viral kinetics of HERV-K in HIV-1-infected patients receiving highly active antiretroviral therapy (HAART) is not unknown. HERV-K kinetic modeling may provide insight into factors altering the effectiveness of HAART in suppressing HIV-1 burden. We conducted a longitudinal study measuring the HERV-K RNA titers in four patients with successful HIV-1-suppressive HAART and in six patients undergoing HAART failure. HERV-K titers were usually undetectable in patients with successful HAART, and when detected, HERV-K titers remained below 5000 copies/ml. On the other hand, HERV-K RNA was consistently detected in patients who failed to respond to HAART before and after HIV-1 rebounds (p < 0.001). Elevated HERV-K RNA titers frequently preceded HIV-1 rebounds. These results suggest that HERV-K viral load may predict HIV-1 reactivation. HERV-K RNA testing might be clinically useful in predicting the onset of HIV-1 resistance due to suboptimal antiretroviral drug levels and/or poor adherence to treatment. PMID- 17919103 TI - Characterization of the full-length human immunodeficiency virus-1 genome from recently infected subjects in Brazil. PMID- 17919104 TI - HIV-1 variants from a perinatal transmission pair demonstrate similar genetic and replicative properties in tonsillar tissues and peripheral blood mononuclear cells. AB - Human immunodeficiency virus type 1 (HIV-1) can be acquired through oropharyngeal tissues in breastfeeding infants. Efforts to better understand the determinants of breast milk transmission are hampered by the lack of a relevant oral human mucosa model and well-defined breast milk-derived viruses. This study used human ex vivo palatine tonsil tissues and peripheral blood mononuclear cells (PBMCs) to characterize the genetic, biological, and replicative properties of HIV-1 variants obtained from a perinatal transmission pair. Unique viral populations from maternal breast milk and infant blood were identified by gp120 V1-V2- and V3 specific heteroduplex tracking assays (HTAs). Full-length infectious recombinant viruses, containing a common HIV-1 NL4-3 genetic background, were generated with V1-V3 gp120 fragments from maternal and infant isolates representing the major viral populations identified in the HTAs. The resulting recombinant viruses used the CCR5 coreceptor, were nonsyncytium forming, and demonstrated replication properties similar to those of parental and control viruses in PBMCs and tonsillar explants. These findings indicate that viruses from breast milk cells and infant blood can infect PBMCs and tonsil tissues. The maternal and infant HIV 1 viruses detailed here will provide useful tools for defining the viral and host factors that contribute to HIV breastfeeding transmission. PMID- 17919105 TI - Interruption of antiretroviral therapy initiated during primary HIV-1 infection: impact of a therapeutic vaccination strategy combined with interleukin (IL)-2 compared with IL-2 alone in the ANRS 095 Randomized Study. AB - HIV-specific T cell responses play a critical role in the control of infection. We evaluated the impact of immune-based interventions in patients first treated during primary HIV-1 infection (PHI). Forty-three patients were randomized within three groups, to receive either interleukin-2 (IL-2 group), or boosts of ALVAC HIV (vCP1433) and LIPO-6T followed by interleukin-2 (Vac-IL2 group), compared with no immune intervention (control group), and were monitored for T cell responses. Impact of strategies on viral replication was subsequently assessed during long-term treatment interruption. HIV-specific CD4(+) T cell responses did not change during the study period in immunized patients relative to controls, and vaccination had only a transient effect on interferon-gamma-producing CD8 responses. Viral rebound after treatment interruption was similar in immunized patients and controls. Forty percent of patients had HIV RNA values <10,000 copies/ml 12 weeks after interruption. The cumulative time off treatment represented almost half the total follow-up period. Immunological and virological status during PHI and HIV DNA load at interruption were predictive of the level of viral rebound after treatment interruption, whereas HIV RNA level during PHI and HIV DNA level at interruption were predictive of the time off treatment. Treatment interruption is safe in patients treated early after primary HIV infection. On the basis of this pilot study, HIV immunizations and interleukin-2 appear to have no supplementary benefit. PMID- 17919106 TI - Sequence diversity among chimpanzee simian immunodeficiency viruses (SIVcpz) suggests that SIVcpzPts was derived from SIVcpzPtt through additional recombination events. AB - Only two of four chimpanzee subspecies (Pan troglodytes), P. troglodytes troglodytes (P.t.t.) and P. troglodytes schweinfurthii (P.t.s.), appear to carry specific simian immunodeficiency viruses (SIVs). We identified genomic features indicating that SIVcpzPtt and SIVcpzPts may have partly different evolutionary histories. A maximum likelihood test to discriminate between hypotheses of a common versus separate origin of SIVcpzPtt and SIVcpzPts revealed three putative regions of separate histories. Thus, after the P.t.t. and P.t.s. split, SIV superinfection led to further recombination resulting in the emergence of SIVcpzPts. This shows that there have been multiple SIV transfers to chimpanzees at different times in their evolution. PMID- 17919107 TI - Characterization of HIV-1 isolates from antiretroviral drug-naive children in southern India. AB - Access to antiretroviral therapy has expanded in many developing countries, including India. The standard first-line regimens consist of a combination of two nucleoside reverse transcriptase inhibitors and a nonnucleoside reverse transcriptase inhibitor, in a fixed drug combination. Data regarding resistance to these drugs are scarce, especially in children. We evaluated the pattern of polymorphism and potential drug resistance mutations (DRMs) in HIV-1 isolates from 48 children naive to antiretroviral therapy attending the outpatient clinics of the Tuberculosis Research Center in Chennai. The samples were subjected to genotyping of reverse transcriptase (RT) and protease genes. All the samples showed significant polymorphisms in both RT and protease genes, but none had major DRMs. The currently recommended generic first-line antiretroviral drug combination is an appropriate treatment strategy for HIV-1-infected children in India. PMID- 17919109 TI - Genetic diversity of HIV-1 non-B strains in Sicily: evidence of intersubtype recombinants by sequence analysis of gag, pol, and env genes. AB - The molecular epidemiology of HIV-1 strains in Sicily (Italy) was phylogenetically investigated by the analysis of HIV-1 gag, pol, and env gene sequences from 11 HIV-1 non-B strains from 408 HIV-1-seropositive patients observed from September 2001 to August 2006. Sequences suggestive of recombination were further investigated by bootscanning analysis of various fragments. Overall, we identified several second-generation recombinant (SGRs) strains, which contained genetic material of CRF02_AG in at least one gene. Notably, three individuals were found to be infected with subsubtype A3, and one of them showed genetic recombination with subsubtype A4. The current study emphasizes the genetic analysis of gag, pol, and env genes as a powerful tool to trace the spread of complex HIV-1 recombinant forms, and highlight the genetic diversity of HIV-1 non-B strains in Italy. PMID- 17919110 TI - Near full-length sequence analysis of a Unique CRF01_AE/B recombinant from Kuala Lumpur, Malaysia. AB - A new HIV-1 circulating recombinant form (CRF), CRF33_01B, has been identified in Malaysia. Concurrently we found a unique recombinant form (URF), that is, the HIV 1 isolate 06MYKLD46, in Kuala Lumpur, Malaysia. It is composed of B or a Thai variant of the B subtype (B') and CRF01_AE. Here, we determined the near full length genome of the isolate 06MYKLD46 and performed detailed phylogenetic and bootscanning analyses to characterize its mosaic composition and to further confirm the subtype assignments. Although the majority of the 06MYKLD46 genome is CRF01_AE, we found three short fragments of B or B' subtype inserted along the genome. These B or B' subtype regions were 716 and 335 bp, respectively, in the protease-reverse transcriptase (PR-RT) region, similar to those found in CRF33_01B, as well as an extra 590 bp in the env gene region. Thus we suggest that 06MYKLD46 is a possible second-generation HIV-1 recombinant derived from CRF33_01B. PMID- 17919108 TI - HTLV-1a tax gene and long terminal repeat sequences from Argentinean strains reveal disagreement with tax restriction fragment length polymorphism subtyping. AB - Sequence and cluster analysis have shown two HTLV-1a tax gene subgroups, tax A and tax B, which are related to long terminal repeat (LTR) molecular subtypes. On the basis of subgroup-specific nucleotide substitutions, restriction fragment length polymorphism (RFLP) analysis of the tax gene for subtyping HTLV-1a isolates was proposed. In this study we genetically characterized the tax gene from 63 HTLV-1-positive Argentinean individuals, including 14 patients with HTLV 1-associated myelopathy/tropical spastic paraparesis and 49 healthy HTLV-1 carriers. RFLP analysis showed that 48 samples yielded the tax A profile (76.19%) and that 15 samples contained the uncut tax B profile (23.81%). However, the LTR and tax sequence analysis revealed that in fact only 2 from the 15 samples belonged to the HTLV-1aB subgroup, presenting four tax B subgroup-specific nucleotide substitutions. The tax gene cluster analysis also confirmed that the majority of Argentinean strains belonged to the Transcontinental HTLV-1aA subgroup. These results indicate that the tax gene RFLP assay which has been proposed and used by some authors to screen HTLV-1a subgroups, is not a suitable tool to perform molecular epidemiological characterization of HTLV-1a populations. PMID- 17919111 TI - Phylogeny of human T cell lymphotropic virus type 1 in Peru: a high degree of evolutionary relatedness with South African isolates. AB - We investigated the prevalence and molecular epidemiology of human T cell lymphotropic virus type 1 in Peruvian HIV-1-positive subjects, and found a 10.1% prevalence in a consecutive series of 318 HIV-1-positive patients living in Lima. Phylogenetic analysis of the long terminal repeat of 10 patient isolates showed that all of them belonged to the HTLV-1aA (Transcontinental) subgroup. Although the majority of the Peruvian sequences included in the analysis formed a clade with other Latin American sequences, the isolates of three patients clustered significantly with South African strains. These data show a high prevalence of HTLV-1 infection in HIV-1-positive subjects living in Lima and confirm the presence in Latin America of HTLV-1 strains probably arising from South Africa. PMID- 17919112 TI - Lack of differences in HIV-1 nef functional domains in infected chinese blood donors at different stages of disease. AB - HIV-1 nef regions were amplified by polymerase chain reaction and sequenced from DNA samples of five asymptomatic subjects and five AIDS patients from a cohort of HIV-1-infected Chinese plasma and blood donors. Sequence analysis revealed that regardless of the stage of disease, each patient's HIV-1 nef sequences belonged to the clade B' subtype. Although there are some differences between the sequences from different patients, no significant differences have been detected in nef nucleotide sequences or functional motifs in the deduced amino acid sequences from patients at different stages of the disease. Furthermore, the predicted binding motifs of HLA-A2 and HLA-A11 were highly conserved among patient nef sequences. These results will contribute to a better understanding of the pathogenesis of circulating HIV-1 in infected Chinese former blood donors and may have important implications in developing an epitope-based vaccine suitable for Chinese blood donors. PMID- 17919113 TI - HIV-1 antiretroviral drug resistance in recently infected patients in Abidjan, Cote d'Ivoire: A 4-year survey, 2002-2006. AB - We performed HIV-1 drug resistance genotypic analysis of viral isolates from 100 antiretroviral (ARV)-naive, recently HIV-1-infected (between 2002 and 2006) individuals from Abidjan (Cote d'Ivoire). The overall prevalence of HIV-1 variants with resistance mutations to reverse transcriptase, protease, or fusion inhibitors was 6%. The majority of isolates were CRF02_AG. Compared with a previous study carried out by our group in 2001-2002 in a similar population in Abidjan, our findings confirm the circulation and transmission of HIV-1 carrying key ARV drug resistance mutation. PMID- 17919114 TI - Resistin: a hormone which induces insulin resistance is increased in normal pregnancy. AB - AIMS: Resistin, a newly discovered adipokine, is thought to play a key role in the regulation of insulin resistance. The objectives of this study were to develop a nomogram of maternal plasma concentrations of resistin from 11 weeks of gestation to term and to determine whether resistin concentrations differ between normal and overweight pregnant women. METHODS: In this cross-sectional study, plasma concentrations of resistin were determined in normal pregnant women of normal body mass index (BMI 18.5-24.9; n=261), overweight pregnant women (BMI > or =25; n=140), and non-pregnant women of normal BMI (n=40). Blood samples were collected once from each woman between the first trimester and term. Percentiles for resistin concentration were determined for five pre-specified windows of gestational age. Plasma resistin concentration was determined by immunoassay. Non parametric statistics were used for analysis. RESULTS: The median maternal plasma concentration of resistin between 11 to 14 weeks of gestation in women of normal weight was significantly higher than non-pregnant women; the plasma concentration of resistin increased with gestational age. CONCLUSIONS: Normal pregnant women have a higher median plasma concentration of resistin than non-pregnant women and the concentration of this adipokine increases with advancing gestation. Alterations in the maternal plasma concentration of resistin during pregnancy could contribute to metabolic changes of pregnancy. PMID- 17919117 TI - A gain-of-function mutant of Munc18-1 stimulates secretory granule recruitment and exocytosis and reveals a direct interaction of Munc18-1 with Rab3. AB - Munc18-1 plays a crucial role in regulated exocytosis in neurons and neuroendocrine cells through modulation of vesicle docking and membrane fusion. The molecular basis for Munc18 function is still unclear, as are the links with Rabs and SNARE [SNAP (soluble N-ethylmaleimide-sensitive factor-attachment protein) receptor] proteins that are also required. Munc18-1 can bind to SNAREs through at least three modes of interaction, including binding to the closed conformation of syntaxin 1. Using a gain-of-function mutant of Munc18-1 (E466K), which is based on a mutation in the related yeast protein Sly1p, we have identified a direct interaction of Munc18-1 with Rab3A, which is increased by the mutation. Expression of Munc18-1 with the E466K mutation increased exocytosis in adrenal chromaffin cells and PC12 cells (pheochromocytoma cells) and was found to increase the density of secretory granules at the periphery of PC12 cells, suggesting a stimulatory effect on granule recruitment through docking or tethering. Both the increase in exocytosis and changes in granule distribution appear to require Munc18-1 E466K binding to the closed form of syntaxin 1, suggesting a role for this interaction in bridging Rab- and SNARE-mediated events in exocytosis. PMID- 17919118 TI - Chelator-facilitated removal of iron from transferrin: relevance to combined chelation therapy. AB - Current iron chelation therapy consists primarily of DFO (desferrioxamine), which has to be administered via intravenous infusion, together with deferiprone and deferasirox, which are orally-active chelators. These chelators, although effective at decreasing the iron load, are associated with a number of side effects. Grady suggested that the combined administration of a smaller bidentate chelator and a larger hexadentate chelator, such as DFO, would result in greater iron removal than either chelator alone [Grady, Bardoukas and Giardina (1998) Blood 92, 16b]. This in turn could lead to a decrease in the chelator dose required. To test this hypothesis, the rate of iron transfer from a range of bidentate HPO (hydroxypyridin-4-one) chelators to DFO was monitored. Spectroscopic methods were utilized to monitor the decrease in the concentration of the Fe-HPO complex. Having established that the shuttling of iron from the bidentate chelator to DFO does occur under clinically relevant concentrations of chelator, studies were undertaken to evaluate whether this mechanism of transfer would apply to iron removal from transferrin. Again, the simultaneous presence of both a bidentate chelator and DFO was found to enhance the rate of iron chelation from transferrin at clinically relevant chelator levels. Deferiprone was found to be particularly effective at 'shuttling' iron from transferrin to DFO, probably as a result of its small size and relative low affinity for iron compared with other analogous HPO chelators. PMID- 17919115 TI - Adiponectin in severe preeclampsia. AB - AIMS: Adiponectin is an adipokine with insulin-sensitizing, anti-atherogenic, anti-inflammatory and angiogenic properties. The aims of this study were to determine whether maternal plasma adiponectin concentrations differ between patients with severe preeclampsia and those with normal pregnancies, and to explore the relationship between plasma adiponectin and the results of Doppler velocimetry of the uterine arteries. METHODS: This case-control study included two groups: (1) patients with severe preeclampsia (n=50) and (2) patients with normal pregnancies (n=150). Pulsed-wave and color Doppler ultrasound examination of the uterine arteries were performed. Plasma adiponectin concentrations were determined by ELISA. Non-parametric statistics were used for analysis. RESULTS: (1) Patients with severe preeclampsia had a higher median plasma concentration of adiponectin than that of normal pregnant women. (2) The median plasma adiponectin concentration did not differ between women with severe preeclampsia who had a high impedance to blood flow in the uterine arteries and those with normal impedance to blood flow. (3) Among patients with normal pregnancies, plasma adiponectin concentrations were negatively correlated with BMI in the first trimester and at sampling. CONCLUSIONS: Women with severe preeclampsia have a higher median plasma concentration of adiponectin than that of normal pregnant women. This may reflect a compensatory feedback mechanism to the metabolically altered, anti-angiogenic and pro-atherogenic state of severe preeclampsia. PMID- 17919119 TI - Mechanisms of endoplasmic-reticulum export of glycine transporter-1 (GLYT1). AB - The GLYT1 (glycine transporter-1) regulates both glycinergic and glutamatergic neurotransmission by controlling the reuptake of glycine at synapses. Trafficking to the cell surface of GLYT1 is critical for its function. In the present paper, by using mutational analysis of the GLYT1 C-terminal domain, we identified the evolutionarily conserved motif R(575)L(576)(X(8))D(585) as being necessary for ER (endoplasmic reticulum) export. This is probably due to its capacity to bind Sec24D, a component of the COPII (coatomer coat protein II) complex. This ER export motif was active when introduced into the related GLYT2 transporter but not in the unrelated VSVG (vesicular-stomatitis virus glycoprotein)-GLYT1 protein in which this motif was mutated but was not transported to the plasma membrane, although this effect was rescued by co-expressing these mutants with wild-type GLYT1. This behaviour suggests that GLYT1 might form oligomers along the trafficking pathway. Cross-linking assays performed in rat brain synaptosomes and FRET (fluorescence resonance energy transfer) microscopy in living cells confirmed the existence of GLYT1 oligomers. In summary, we have identified a motif involved in the ER exit of GLYT1 and, in analysing the influence of this motif, we have found evidence that oligomerization is important for the trafficking of GLYT1 to the cell surface. Because this motif is conserved in the NSS (sodium- and chloride-dependent neurotransmitter transporter) family, it is possible that this finding could be extrapolated to other related transporters. PMID- 17919116 TI - Plasma adiponectin concentrations in non-pregnant, normal and overweight pregnant women. AB - AIMS: Adiponectin is an adipokine that has anti-diabetic, anti-atherogenic, anti inflammatory and angiogenic properties. This hormone has been implicated in both the physiological adaptation to normal pregnancy and in obstetrical complications. The aims of this study were to determine normal maternal plasma concentrations of adiponectin throughout gestation and to explore the relationships between plasma adiponectin concentration, pregnancy, and maternal overweight. METHODS: A cross-sectional study was designed to include normal pregnant (normal weight and overweight; 11-42 weeks of gestation), and non pregnant women. Plasma adiponectin concentration was determined by immunoassay. Non-parametric statistics were used for analysis. RESULTS: (1) Adiponectin was detectable in the plasma of all patients; (2) there was no significant differences in the median adiponectin concentration between pregnant and non pregnant women; (3) plasma adiponectin concentrations were negatively correlated with gestational age only among normal weight pregnant women; and (4) overweight patients had significantly lower plasma adiponectin concentrations than normal weight women. CONCLUSIONS: Consistent with the increased insulin resistance and weight gain that occur in pregnancy, adiponectin concentrations were negatively correlated with gestational age. The results of this study and the nomogram herein presented, can serve as the basis to explore the relationship between adiponectin and pregnancy complications and facilitate the clinical use of this important adipokine. PMID- 17919120 TI - Novel molecules for intra-oral delivery of antimicrobials to prevent and treat oral infectious diseases. AB - New molecules were designed for efficient intra-oral delivery of antimicrobials to prevent and treat oral infection. The salivary statherin fragment, which has high affinity for the tooth enamel, was used as a carrier peptide. This was linked through the side chain of the N-terminal residue to the C-terminus of a defensin-like 12-residue peptide to generate two bifunctional hybrid molecules, one with an ester linkage and the other with an anhydride bond between the carrier and the antimicrobial components. They were examined for their affinity to a HAP (hydroxyapatite) surface. The extent of the antimicrobial release in human whole saliva was determined using 13C-NMR spectroscopy. The candidacidal activity of the molecules was determined as a function of the antimicrobial release from the carrier peptide in human saliva. The hybrid-adsorbed HAP surface was examined against Candida albicans and Aggregatibacter actinomycetemcomitans using the fluorescence technique. The bifunctional molecules were tested on human erythrocytes, GECs (gingival epithelial cells) and GFCs (gingival fibroblast cells) for cytotoxicity. They were found to possess high affinity for the HAP mineral. In human whole saliva, a sustained antimicrobial release over a period of more than 40-60 h, and candidacidal activity consistent with the extent of hybrid dissociation were observed. Moreover, the bifunctional peptide-bound HAP surface was found to exhibit antimicrobial activity when suspended in clarified human saliva. The hybrid peptides did not show any toxic influence on human erythrocytes, GECs and GFCs. These novel hybrids could be safely used to deliver therapeutic agents intra-orally for the treatment and prevention of oral infectious diseases. PMID- 17919121 TI - Polyamines modulate the subcellular localization of RNA-binding protein HuR through AMP-activated protein kinase-regulated phosphorylation and acetylation of importin alpha1. AB - Polyamines are required for maintenance of intestinal epithelial integrity, and a decrease in cellular polyamines increases the cytoplasmic levels of RNA-binding protein HuR stabilizing p53 and nucleophosmin mRNAs, thus inhibiting IEC (intestinal epithelial cell) proliferation. The AMPK (AMP-activated protein kinase), an enzyme involved in responding to metabolic stress, was recently found to be implicated in regulating the nuclear import of HuR. Here, we provide evidence showing that polyamines modulate subcellular localization of HuR through AMPK-regulated phosphorylation and acetylation of Impalpha1 (importin alpha1) in IECs. Decreased levels of cellular polyamines as a result of inhibiting ODC (ornithine decarboxylase) with DFMO (D,L-alpha-difluoromethylornithine) repressed AMPK activity and reduced Impalpha1 levels, whereas increased levels of polyamines as a result of ODC overexpression induced both AMPK and Impalpha1 levels. AMPK activation by overexpression of the AMPK gene increased Impalpha1 but reduced the cytoplasmic levels of HuR in control and polyamine-deficient cells. IECs overexpressing wild-type Impalpha1 exhibited a decrease in cytoplasmic HuR abundance, while cells overexpressing Impalpha1 proteins bearing K22R (lacking acetylation site), S105A (lacking phosphorylation site) or K22R/S105A (lacking both sites) mutations displayed increased levels of cytoplasmic HuR. Ectopic expression of these Impalpha1 mutants also prevented the increased levels of cytoplasmic HuR following polyamine depletion. These results indicate that polyamine-mediated AMPK activation triggers HuR nuclear import through phosphorylation and acetylation of Impalpha1 in IECs and that polyamine depletion increases cytoplasmic levels of HuR as a result of inactivation of the AMPK-driven Impalpha1 pathway. PMID- 17919122 TI - Repair of osteochondral defects with a construct of mesenchymal stem cells and a polydioxanone/poly(vinyl alcohol) scaffold. AB - The purpose of the present study was to test the potential of an MSC (mesenchymal stem cell)/PDO/PVA [polydioxanone/poly(vinyl alcohol)] hybrid scaffold construct to repair cartilage defects. For the in vitro study, MSCs were isolated from human bone marrow and cultured in PDO/PVA scaffolds for 4 weeks. Gross and microscopic examinations were performed, as well as RT-PCR (reverse-transcription PCR) analyses for cartilage-specific genes. For the in vivo study, MSCs isolated from rabbits were cultured in the PDO/PVA scaffolds and tissue-engineered into neocartilage, then implanted into the osteochondral defect on the distal femur of the same rabbits. Gross and histological evaluations were performed at 8 weeks after the implantation. The results of the in vitro study demonstrated that the physical stability of the cell-cultured hybrid scaffold was maintained until 4 weeks after initial placement. Scanning electronmicroscopy indicated the infiltration of the cells into, and appropriate interactions with, the scaffolds. RT-PCR showed an expression of cartilage-specific genes similar to that seen with pellet-cultured MSCs. From the in vivo study, the defect area of the experimental group showed smooth consistent glistening white tissue resembling articular cartilage, whereas the control group showed a red irregular tissue with surface depression. The regenerated cartilage of the experimental group showed metachromasia on both Safranin-O and dense staining for type II collagen, whereas the control group showed little metachromatic staining and less intense staining for type II collagen. A histological score of the quality of cartilage formation indicated that the MSC/PDO/PVA hybrid scaffold successfully produced neocartilage in vitro and enhanced the repair of the osteochondral defect in vivo. Further in vivo studies will be necessary to elucidate further the value of PDO/PVA as a scaffold material for cartilage regeneration. PMID- 17919123 TI - Direct cardiac actions of erythropoietin (EPO): effects on cardiac contractility, BNP secretion and ischaemia/reperfusion injury. AB - EPO (erythropoietin) has recently been shown to have protective actions upon the myocardium; however, the direct effects of EPO upon cardiac contractile and secretory functions are unknown and the signalling mechanisms are not well defined. In the present study, we provide the first evidence of direct cardiac contractile actions of EPO. In isolated perfused Sprague-Dawley rat hearts, a 30 min infusion of EPO significantly increased contractility in a dose-dependent fashion (maximal change 18+/-2% with 1 unit/ml EPO; P<0.005 compared with vehicle). Perfusate ET-1 (endothelin-1) increased transiently during EPO infusion, and the ET(A/)ET(B) antagonist bosentan abolished the inotropic response to EPO. BNP (B-type natriuretic peptide) secretion (28+/-8%; P<0.05) and nuclear transcription factor GATA-4 DNA-binding activity (51%; P<0.05) were both significantly increased by EPO and blocked by bosentan. In a model of global ischaemic injury, delivery of 1 unit/ml EPO during reperfusion significantly attenuated creatine kinase release (28+/-12%; P<0.05) and significantly improved contractile recovery (P<0.001), independent of ET(A) blockade. Apoptotic indices [assessed by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling)/cleaved caspase-3-positive cells] were significantly decreased (P<0.01) by 1 unit/ml EPO during reperfusion alone, coincident with significantly increased phosphorylation of myocardial JAK2 (Janus kinase 2) and STAT3 (signal transducer and activator of transcription 3). Thus EPO directly enhances cardiac contractility and BNP secretion and alleviates ischemia/reperfusion injury via ET 1-dependent and -independent mechanisms respectively. PMID- 17919124 TI - Derivation of the distensibility coefficient using tissue Doppler as a marker of arterial function. AB - To date, the main cardiovascular application of TDI (tissue Doppler imaging) has been in myocardial evaluation. In the present study, we investigated the feasibility and reproducibility of assessing arterial elasticity using the DC (distensibility coefficient) measured by TDI, the correlation of this with the DC obtained by other methods and the DC in patients with various degrees of cardiovascular risk. We studied 450 subjects (256 men; age, 51+/-10 years) with and without risk factors of cardiovascular disease. Arterial displacement was measured from TDI, and B-mode and M-mode images of the common carotid artery in the longitudinal plane, and the DC with each method was compared. Linear regression showed a good correlation between all three methods. The results for TDI and B-mode were comparable [(21+/-10) compared with (21+/-10)x10(-3)/kPa respectively; P=not significant], but there were significant differences between TDI and M-mode [(21+/-10) compared with (31+/-13)x10(-3)/kPa respectively; P<0.0001] and between B-mode and M-mode [(21+/-10) compared with (31+/-13)x10( 3)/kPa respectively; P<0.0001]. Similarly, Bland-Altman analysis showed the least variability in the DC between TDI and B-mode, and there were no significant differences between the average measurements. The TDI DC also had the lowest paired difference for inter-observer variability [(-0.1+/-1.1)x10(-3)/kPa; P=not significant]. In conclusion, the results of the present study suggest that TDI of the carotid arteries is feasible, comparable with B-mode measurements, more robust than M-mode and less variable than the other methods. PMID- 17919126 TI - The role of the neurobiologist in redefining the diagnosis of autism. AB - Until recently, autism, along with the other developmental disabilities, was largely ignored by the medical and research community. At this early point in our understanding of the syndrome, neurobiologists and especially those who work with human brain tissue have a great deal to offer. A thorough understanding of the clinically defined syndrome is essential. Along with the other psychiatric diseases listed in the Diagnostic and Statistical Manual of Mental Disorders (DSM), autism is defined by gross behavioral macros that, in all probability, are only indirectly related to basic biological systems. The diagnostic schema is not etiologically based. The diagnostic triad of symptoms that defines autism- impaired communication, impaired social interaction, and restricted and repetitive interests and activities--has been found to be present in the general population with no clear demarcation between pathological severity and being a common trait. In addition, the three basic symptoms of autism appear not to associate highly, thus leaving undetermined the validity of studying autism in its currently defined triad of symptoms. It is proposed that a close working relationship between neurobiologists and clinicians is necessary in order to identify etiologically based diagnostic schemas that would complement, rather than replace, the clinical diagnosis. PMID- 17919127 TI - Autism brain tissue banking. AB - One avenue of progress toward understanding the neurobiological basis of autism is through the detailed study of the post-mortem brain from affected individuals. The primary purpose of autism brain tissue banking is to make well-characterized and optimally preserved post-mortem brain tissue available to the neuroscience research community. In this paper we discuss our current understanding of the criteria for optimal characterization and preservation of post-mortem brain tissue; the pitfalls associated with inadequate clinical and neuropathological characterization and the advantages and disadvantages of post-mortem studies of the brain. We then describe the current status of the brain tissue bank supported by the Autism Tissue Program, including the demographic characteristics of the tissue donors, post-mortem interval, sex, age and the method of preservation. Finally, we provide information on the policies and procedures that govern the distribution of brain specimens by this bank and the nature of the studies that are currently being supported directly by this program. PMID- 17919128 TI - The neuropathology of autism. AB - Autism is a brain disorder characterized by abnormalities in how a person relates and communicates to others. Both post-mortem and neuroimaging studies indicate the presence of increased brain volume and, in some cases, an altered gray/white matter ratio. Contrary to established gross findings there is no recognized microscopic pathology to autism. Early studies provided multiple leads none of which have been validated. Clinicopathological associations have been difficult to sustain when considering possible variables such as use of medications, seizures, mental retardation and agonal/pre-agonal conditions. Research findings suggest widespread cortical abnormalities, lack of a vascular component and an intact blood-brain barrier. Many of the previously mentioned findings can be explained in terms of a mini-columnopathy. The significance of future controlled studies should be judged based on their explanatory powers; that is, how well do they relate to brain growth abnormalities and/or provide useful clinicopathological correlates. PMID- 17919129 TI - The neurobiology of autism. AB - Improving clinical tests are allowing us to more precisely classify autism spectrum disorders and diagnose them at earlier ages. This raises the possibility of earlier and potentially more effective therapeutic interventions. To fully capitalize on this opportunity, however, will require better understanding of the neurobiological changes underlying this devastating group of developmental disorders. It is becoming clear that the normal trajectory of neurodevelopment is altered in autism, with aberrations in brain growth, neuronal patterning and cortical connectivity. Changes to the structure and function of synapses and dendrites have also been strongly implicated in the pathology of autism by morphological, genetic and animal modeling studies. Finally, environmental factors are likely to interact with the underlying genetic profile, and foster the clinical heterogeneity seen in autism spectrum disorders. In this review we attempt to link the molecular pathways altered in autism to the neurodevelopmental and clinical changes that characterize the disease. We focus on signaling molecules such as neurotrophin, Reelin, PTEN and hepatocyte growth factor, neurotransmitters such as serotonin and glutamate, and synaptic proteins such as neurexin, SHANK and neuroligin. We also discuss evidence implicating oxidative stress, neuroglial activation and neuroimmunity in autism. PMID- 17919130 TI - Mouse behavioral assays relevant to the symptoms of autism. AB - While the cause of autism remains unknown, the high concordance between monozygotic twins supports a strong genetic component. The importance of genetic factors in autism encourages the development of mutant mouse models, to advance our understanding of biological mechanisms underlying autistic behaviors. Mouse models of human neuropsychiatric diseases are designed to optimize (i) face validity (resemblance to the human symptoms) (ii) construct validity (similarity to the underlying causes of the disease) and (iii) predictive validity (expected responses to treatments that are effective in the human disease). There is a growing need for mouse behavioral tasks with all three types of validity, to define robust phenotypes in mouse models of autism. Ideal mouse models will incorporate analogies to the three diagnostic symptoms of autism: abnormal social interactions, deficits in communication and high levels of repetitive behaviors. Social approach is tested in an automated three chambered apparatus that offers the subject a choice between spending time with another mouse, with a novel object, or remaining in an empty familiar environment. Reciprocal social interaction is scored from videotapes of interactions between pairs of unfamiliar mice. Communication is evaluated by measuring emission and responses to vocalizations and olfactory cues. Repetitive behaviors are scored for measures of grooming, jumping, or stereotyped sniffing of one location or object. Insistence on sameness is modeled by scoring a change in habit, for example, reversal of the spatial location of a reinforcer in the Morris water maze or T-maze. Associated features of autism, for example, mouse phenotypes relevant to anxiety, seizures, sleep disturbances and sensory hypersensitivity, may be useful to include in a mouse model that meets some of the core diagnostic criteria. Applications of these assays include (i) behavioral phenotyping of transgenic and knockout mice with mutations in genes relevant to autism; (ii) characterization of inbred strains of mice; (iii) evaluation of environmental toxins; (iv) comparison of behavioral phenotypes with genetic factors, such as unusual expression patterns of genes or unusual single nucleotide polymorphisms; and (v) evaluation of proposed therapeutics for the treatment of autism. PMID- 17919131 TI - A 57-year-old man with a dural-based parietal lobe tumor. PMID- 17919132 TI - A 12-year-old girl with seizures and dementia. PMID- 17919133 TI - A 63-year-old man with dementia, ataxia and VI nerve palsy. PMID- 17919134 TI - A 15-year-old girl with a cystic brain tumor. PMID- 17919135 TI - Leg weakness after liver transplant. PMID- 17919136 TI - A 55-year-old man with liver failure, delirium and seizures. PMID- 17919137 TI - The European LABDEL project and its relevance to the prevention and treatment of allergies. AB - In March 2001, the European Commission funded a 3-year project (contract no. QLK3 CT-2000-00340) under the fifth Framework Programme to develop and test prototype products based on the oral delivery of vaccine and therapeutic agents using harmless lactic acid bacteria (LAB). The project, best known under its acronym LABDEL (for LAB delivery) also included research on LAB fermentation and technological innovations aimed at enhancing the efficiency of LAB delivery systems (1). One of the key scientific objectives was to investigate the possibility to prevent or treat a type I allergic disease using mucosal administration of LAB expressing the pollen allergen Bet v 1. The aim of this paper was to describe the background of the project with reference to a limited selection of articles and recent reviews as well as the results and major conclusions arising from this part of the project. PMID- 17919138 TI - Strategy for allergenicity assessment of 'natural novel foods': clinical and molecular investigation of exotic vegetables (water spinach, hyacinth bean and Ethiopian eggplant). AB - BACKGROUND: Foods not commonly consumed in the European Union must be proven safe before being brought to market, including an assessment of allergenicity. We present a three-stepwise strategy for allergenicity assessment of natural novel foods using three novel vegetables, namely, water spinach, hyacinth bean, Ethiopian eggplant. METHODS: First, vegetable extracts were analyzed for the presence of pan-allergens [Bet v 1 homologous proteins, profilins, nonspecific lipid transfer proteins (LTP)] by immunoblot analysis with specific animal antibodies. Secondly, the IgE-binding of the food extracts was investigated by EAST (Enzyme-allergosorbent test) and immunoblot analysis using sera with IgE reactivity to known pan-allergens or to phylogenetically related foods from subjects (i) allergic to birch, grass and mugwort pollen, (ii) with food allergy to soy, peanut, tomato, multiple pollen-related foods and (iii) sensitized to LTP. Thirdly, the clinical relevance of IgE-binding was assessed in vivo by skin prick testing (SPT) and open oral food challenges (OFC). RESULTS: Profilin and LTP were detected by animal antibodies in all vegetables, a Bet v 1 homologue selectively in hyacinth bean. IgE-binding to LTP, profilin and a Bet v 1 homologue was proven by immunoblot analysis and EAST. Positive SPT and OFC results were observed for all vegetables in pollen-allergic patients. CONCLUSIONS: Our stepwise procedure confirmed the presence and IgE-binding capacity of novel vegetable proteins homologous to known allergens in endemic vegetable foods. In vivo testing proved the potential of the novel vegetables to elicit clinical allergy. Hence, our described algorithm seems to be applicable for allergenicity testing of natural novel foods. PMID- 17919139 TI - Oat sensitization in children with atopic dermatitis: prevalence, risks and associated factors. AB - BACKGROUND: Topical treatments of atopic dermatitis (AD) may be responsible for cutaneous allergies. Percutaneous sensitization to oat used in emollients/moisturizers has already been reported. Our objectives were to measure the prevalence of oat sensitization in AD children, to assess its relevance, and to look at related parameters. METHODS: We recruited prospectively children with AD referred for allergy testing between June 2001 and December 2004. Atopy patch tests (APT) and skin prick tests (SPT) to oat proteins (1%, 3% and 5%) and to the European standard series were performed followed by oral food challenge (OFC) and repeated open application test (ROAT) in the oat-sensitized group. RESULTS: About 302 children were enrolled. Oat APT and SPT were positive in 14.6% and 19.2% of cases, respectively. Children under 2 years of age were more likely to have positive APT. In oat-sensitized children, OFC and ROAT were positive in 15.6% (five of 32) and 28% (seven of 25) of cases, respectively. Thirty-two percentage of oat cream users had oat-positive atopy patch test (APT) vs 0% in the nonusers group. CONCLUSIONS: Oat sensitization in AD children seen for allergy testing is higher than expected. It may be the result of repeated applications of cosmetics with oats on a predisposed impaired epidermal barrier. We suggest avoiding topical-containing oat proteins in infants with AD. PMID- 17919141 TI - Randomized, placebo-controlled trial of Lactobacillus rhamnosus GG as treatment of atopic dermatitis in infancy. AB - BACKGROUND: Some studies have suggested that supplementation of food with lactobacilli may prevent or improve atopic dermatitis in children. This study was designed to investigate the therapeutic effect of Lactobacillus rhamnosus GG (LGG) as a food supplement in infants suffering from atopic dermatitis. METHODS: Infants aged 3-12 months suffering from mild-to-moderate atopic dermatitis (severity scoring of atopic dermatitis or SCORAD index of 15-40) without current antiinflammatory treatment were randomized to receive LGG (5 x 10(9) colony forming units b.i.d.) or placebo as a food supplement for 12 weeks. Severity scoring of atopic dermatitis index and use of hydrocortisone 1% ointment as rescue medication (2 points per application) were recorded at 4, 8, and 12 weeks of treatment and combined as symptom load (SL). RESULTS: Fifty-four infants (LGG group, mean +/- SD SCORAD index 24.6 +/- 8.8) and 48 infants (placebo group, SCORAD index 23.6 +/- 7.8) were randomized and completed the treatment period (intention-to-treat analysis). Symptom load generally improved over time at 4 weeks (LGG vs placebo, 23.8 +/- 12.4 vs 20.6 +/- 9.9), 8 weeks (22.5 +/- 14.6 vs 17.9 +/- 13.1), and 12 weeks (19.6 +/- 15.4 vs 15.1 +/- 12.1), without statistically significant group differences. When stratified for age, eczema severity or use of rescue medication, no statistically significant group differences, in improvement, were found. No significant group differences were found for the use of rescue medication (0.8 +/- 45.0 g vs 3.5 +/- 29.8 g), increase in mean logarithmic total serum IgE (0.17 +/- 0.30 kU/l vs 0.26 +/- 0.45 kU/l), and newly developed allergic sensitization against hen's egg or cow's milk (18.8%vs 10.0%). CONCLUSION: This placebo-controlled trial showed no therapeutic effect of LGG against mild-to-moderate atopic dermatitis in infancy. PMID- 17919140 TI - Specific oral tolerance induction in food allergy in children: efficacy and clinical patterns of reaction. AB - BACKGROUND: Specific oral tolerance induction (SOTI) seems to be a promising treatment of food allergy. Specific oral tolerance induction and elimination diet were compared with respect to efficacy rate and patterns of clinical reaction. METHODS: Children with challenge proven immunoglobulin E (IgE)-mediated cow's milk (CM) allergy or hen's egg (HE) allergy were randomly assigned to SOTI or elimination diet as a control group. Specific oral tolerance induction treatment was performed at home on a daily basis according to a study protocol with fresh CM or lyophilized HE protein. Re-evaluation of clinically relevant food allergy was performed by food challenge after a median of 21 months. Children in the SOTI group received a secondary elimination diet for 2 months prior to follow-up challenge to evaluate persistence of induced oral tolerance. RESULTS: At follow up challenge, nine of 25 children (36%) showed permanent tolerance in the SOTI group, three of 25 (12%) were tolerant with regular intake and four of 25 (16%) were partial responders. In the control group, seven of 20 children (35%) were tolerant. Allergen-specific immunoglobulin E decreased significantly both in children who developed natural tolerance during the elimination diet (P < 0.05) and in those with SOTI (P < 0.001). CONCLUSIONS: Specific oral tolerance induction seems a valid treatment option for patients with persistent food allergy. Indications may be given if avoidance cannot be guaranteed or for those who are eager to eat the food in question. Advantages of SOTI are the increased threshold dose for allergic reactions and the substantially reduced risk of severe allergic reactions after inadvertent ingestion of the allergen. However, careful monitoring during SOTI is mandatory. PMID- 17919142 TI - Natural rubber latex and chestnut allergy: cross-reactivity or co-sensitization? AB - BACKGROUND: Chestnut and natural rubber latex (NRL) allergy are often associated in the latex-fruit syndrome. AIM OF THE STUDY: To establish whether the concurrent NRL and chestnut IgE antibody reactivity are the results of co sensitization or cross-reactivity. METHODS: Sera from 19 patients with chestnut- and NRL-specific IgE were selected and tested for reactivity with recombinant (r) latex allergens. Cross-reactivity was explored by IgE-inhibition experiments using chestnut or NRL allergens as solid phase on ImmunoCAP. RESULTS: IgE antibodies were detected to rHev b 6.01 (prohevein) in 58% of the sera, to rHev b 5 in 32%, to rHev b 12 in four of 13 sera, to rHev b 7.02 and rHev b 11 in four, and to rHev b 1 in two of 19 sera. rHev b 8-IgE antibodies were found in nine sera (47%), whereas six displayed mono-sensitization to rHev b 8 with regard to our test panel. Three of 16 sera showed IgE to cross-reactive carbohydrate determinants. In most sera recognizing rHev b 5 and/or rHev b 6.01 as major allergens the IgE-reactivity to NRL remained unaffected by chestnut extract and chestnut-IgE remained unaffected by NRL extract. Conversely, in sera with rHev b 8 as dominant allergen IgE-binding to NRL was nearly completely inhibited by chestnut and vice versa. IgE-binding to rHev b 8 was abolished by chestnut extract. CONCLUSIONS: Although patients have concomitant IgE antibody reactivity to chestnut and NRL, cross-reactivity could be demonstrated mainly in those patients with IgE to Hev b 8 (profilin) from NRL. PMID- 17919143 TI - Allergic sensitization to cat in childhood as major predictor of incident respiratory allergy in young adults. AB - BACKGROUND: Little is known on the predictive value of sensitization to specific aeroallergens in children with respect to asthma and hay fever incidence in young adulthood. We followed the incidence of asthma and hay fever in children (mean age 11 years) over 9 years, and analyzed the predictive value of sensitization to five common aeroallergens. METHODS: Three consecutive surveys were conducted in East German school children. Specific IgE antibodies to birch and timothy grass pollen, house dust mite, cat, and cladosporium were measured. In 1207 out of the 2453 children, the 9-year incidence of asthma and hay fever was assessed by reported doctors' diagnoses. For sensitization, diagnostic parameters were determined and logistic regression analyses controlled for relevant confounders. RESULTS: A total of 176/78 incident hay fever/asthma cases occurred equaling a cumulative incidence of 1.93/0.86% per year. Incident asthma was associated with previous sensitization to cat [risk ratio (RR) 3.49, 1.57-7.74] and grass pollen (RR 1.79, 1.01-3.19), whereas incident hay fever was associated with each allergen, with grass pollen (RR 6.00, 4.04-8.90) and cat (RR 5.36, 2.87-9.99) exhibiting the strongest associations. When mutually adjusting for all allergens, sensitization to cat remained significantly associated with asthma and hay fever. The latter was also associated with sensitization to grass pollen. The highest positive predictive values for asthma and hay fever were obtained for cat sensitization (10/49 = 20.4% and 23/49 = 46.9%). CONCLUSIONS: Childhood sensitization to cat and grass pollen predicts the incidence of asthma and hay fever in young adulthood. The predictive capacity differs by allergen and manifestation of atopy. PMID- 17919144 TI - The common allergens in the Churg-Strauss syndrome. AB - BACKGROUND: The asthmatic-prodromal phase of Churg-Strauss syndrome (CSS) is usually considered allergic, but data about the involved allergens are scarce. The aim of our work was to examine the prevalence of allergy in a group of CSS patients and in two control groups of persistent asthmatic subjects selected for eosinophilia >10% [first control group patients (CGP1)] and eosinophils <6% [second control group patients (CGP2)]. METHODS: The respiratory symptoms, and the results of prick test and/or RAST for the common allergens, performed before the vasculitic phase in 51 CSS, were retrospectively evaluated and compared with those of 46 CGP1 and 50 CGP2. RESULTS: 31.4% of CSS vs 67.4% of CGP1 (P = 0.0004) and vs 58.0% CGP2 (P = 0.007) were allergic. The number of subjects with seasonal allergies was lower in CSS vs CGP1 (P = 0.0069) and vs CGP2 (P = 0.0002). The number of perennial allergies was significantly higher in CSS than in both control groups (CSS vs CGP1, P = 0.0108; CSS vs CGP2, P = 0.0079). The subjects allergic to Dermatophagoides were prevalent in CSS vs CGP1 (P = 0.0045) but not vs CGP2. CONCLUSIONS: The evidence of allergy, considered as the demonstration of specific IgE consistent with the clinical history, is present in less than one third of CSS and the higher prevalence of seasonal allergies in the controls disagrees with persistent asthma. Allergy may be only one of several mechanisms triggering exacerbation of asthma or supporting chronic airway inflammation as in asthma in general. Alternatively, unidentified allergens may play a role. PMID- 17919145 TI - Infrequent milk consumption plus being overweight may have great risk for asthma in girls. AB - BACKGROUND: Inadequate milk consumption and being overweight are each associated with asthma symptoms in children. Milk consumption has been inversely related to childhood overweight. Thus, being overweight may be a pathway or a confounder in milk-asthma relationship. A combination of both factors may be associated with greater risk of asthma than one factor alone. METHODS: This study included 246 children with allergist-diagnosed asthma and 477 nonasthmatic controls at the age of 8-10 years. Information on milk consumption during the last 12 months was obtained from questionnaires. Being overweight was defined as body mass index > or =85th percentile. Being overweight as a possible pathway or confounder was formally tested. The odds ratio (OR) for combined infrequent milk consumption and being overweight in asthmatic vs nonasthmatic children was determined in multivariate logistic regression analyses. RESULTS: There was a significant interaction between milk consumption and sex of the child in relation to asthma. Asthma was significantly associated with infrequent milk consumption in girls (crude OR 2.11, 95% CI 1.05-4.24) but not in boys. The asthma-milk relation in girls was neither mediated by being overweight (p(mediation tests) = 0.27) nor was the relationship confounded by being overweight (a 9% change in coefficient). Asthmatic girls had 3.6 times increased odds of having combination of infrequent milk consumption and being overweight than nonasthmatic girls (adjusted OR 3.64, 95% CI 1.18-11.24). Asthma was not associated with either factor or with absence of the other in girls. CONCLUSION: Infrequent milk consumption plus being overweight may have great risk for asthma in girls. PMID- 17919146 TI - Interactions between mature Der p 1 and its free prodomain indicate membership of a new family of C1 peptidases. AB - BACKGROUND: Studies in vivo have shown that the cysteine peptidase activity of group 1 house dust mite allergens contributes to their allergenicity. These allergens are synthesized initially as proenzymes and removal of the propiece is necessary to unmask their proteolytic activity. In related C1 family cysteine peptidases of enzyme clan CA, liberated propieces continue to inhibit the mature peptidase as tight binding inhibitors. As it is not known whether mite peptidase allergens behave similarly, our objective was to investigate the effect of the Der p 1 propiece on the catalytic activity of Der p 1 and Der f 1. METHODS: Enzymatic activity of natural Der p 1 and Der f 1 was assessed using a specific substrate and the effect of the recombinant propiece on its enzyme kinetics defined. The integrity of the propiece during these interactions was studied functionally and by analysis of the reaction mixtures. RESULTS: Der p 1 propiece was a potent competitive inhibitor of Der p 1 and Der f 1. In contrast to other cysteine peptidase prodomains, which are cognate tight binding inhibitors, the Der p 1 propiece behaves as a substrate and is fully degraded during this interaction. CONCLUSION: Mature Der p 1-prodomain interactions differ from other C1 family cysteine peptidases, suggesting that group 1 mite allergens are a new subgroup among C1 family cysteine peptidases. The rapid inactivation of Der p 1 prodomain is a newly identified mechanism that may contribute to the potency of this allergen. PMID- 17919147 TI - Allergen induced gene expression of airway epithelial cells shows a possible role for TNF-alpha. AB - BACKGROUND: Epithelium is more than a physical barrier for pathogens and allergens, as it is also capable of producing mediators in response to these environmental factors. Some of these mediators have an immuno-modulatory function, suggesting that epithelium is an active component of the immune response. Here, we fully characterize the expression profile of airway epithelial cells in response to house dust mite (HDM) allergen. METHODS: H292 cells were exposed to HDM extract for 24 h, RNA and supernatant was used for microarray analysis and multiplex enzyme-linked immunosorbent assay (ELISA) respectively. RESULTS: A total of 38,500 genes, 813 were differentially expressed by more than twofold and 116 even more than fivefold. Interestingly, among the most up regulated genes, a large number are involved in cell-to-cell communication. These include chemokines (CCL-8 and -20, CXCL-1, -2 and -3), cytokines (IL-1alpha, -6 and -11), anti-inflammatory factors [PTX-3, interleukin (IL)-13Ralpha, tumour necrosis factor (TNF)-alphaIP3], and factors that are involved in repair of the mucosal tissue (LOXL-2, NID-2, HBEGF, MUC-5AC and MUC-5B). Pathway analysis showed that a number of these genes are transcriptionally regulated by TNF-alpha, which we could detect by quantitative polymerase chain reaction at earlier time points after HDM exposure. In addition, we could detect increased protein levels for TNF-alpha, IL-6, IL8, granulocyte-macrophage colony-stimulating factor, granulocyte colony stimulating factor and interferon (IFN)-gamma using ELISA. CONCLUSION: Our data show that a broad range of mediators produced upon allergen exposure by these mediators' epithelial cells can participate in the immune response via recruitment and activation of cells of the immune system. PMID- 17919148 TI - Lack of dehydroepiandrosterone in type I and II hereditary angioedema and role of danazol in steroid hormone conversion. AB - BACKGROUND: Hereditary angioedema (HAE) is successfully treated with danazol, a therapeutic steroid compound. To investigate hormones of the hypothalamic pituitary-adrenal (HPA) and hypothalamic-pituitary-gonadal (HPG) axis in patients with HAE with and without danazol. METHODS: We included 16 patients with type I HAE, nine patients with type II HAE, and 16 healthy subjects. Serum levels of adrenocorticotropic hormone (ACTH), cortisol, androstenedione, dehydroepiandrosterone (DHEA), free testosterone, and 17beta-oestradiol were measured. RESULTS: Serum levels of ACTH were markedly decreased in patients with type II HAE compared to the other groups (P < 0.001). Serum cortisol was similar between groups but danazol treatment decreased cortisol levels, particularly in women (P = 0.019). Serum levels of DHEA were significantly decreased in all patients with type I and II HAE compared to controls (P < 0.05), which was only partly dependent on prior danazol therapy as patients without danazol had also decreased serum levels of DHEA (P < 0.05). Furthermore, free testosterone serum levels were markedly increased in patients under danazol (P < 0.005) and the ratio of 17beta-oestradiol/free testosterone was significantly decreased in these patients (P < 0.005). CONCLUSIONS: This study demonstrated decreased DHEA in patients with type I and II HAE independent of danazol therapy, which was particularly evident in women. It also demonstrates that danazol induced a marked up-regulation of free testosterone in relation to precursors and downstream 17beta-oestradiol. In HAE, there seems to be a primary lack of the adrenal androgen DHEA. PMID- 17919149 TI - Localized eosinophilic ileitis with mastocytosis successfully treated with oral budesonide. PMID- 17919150 TI - Angioedema after administration of methylprednisolone to treat drug allergy. PMID- 17919151 TI - Airborne occupational allergic contact dermatitis from coal dust. PMID- 17919152 TI - Madness on the streets. PMID- 17919153 TI - Physical illness and schizophrenia: a review of the literature. AB - OBJECTIVE: The lifespan of people with schizophrenia is shortened compared to the general population. We reviewed the literature on comorbid physical diseases in schizophrenia to provide a basis for initiatives to fight this unacceptable situation. METHOD: We searched MEDLINE (1966 - May 2006) combining the MeSH term of schizophrenia with the 23 MeSH terms of general physical disease categories to identify relevant epidemiological studies. RESULTS: A total of 44 202 abstracts were screened. People with schizophrenia have higher prevalences of HIV infection and hepatitis, osteoporosis, altered pain sensitivity, sexual dysfunction, obstetric complications, cardiovascular diseases, overweight, diabetes, dental problems, and polydipsia than the general population. Rheumatoid arthritis and cancer may occur less frequently than in the general population. Eighty-six per cent of the studies came from industrialized countries limiting the generalizability of the findings. CONCLUSION: The increased frequency of physical diseases in schizophrenia might be on account of factors related to schizophrenia and its treatment, but undoubtedly also results from the unsatisfactory organization of health services, from the attitudes of medical doctors, and the social stigma ascribed to the schizophrenic patients. PMID- 17919154 TI - The role of mental disorder in attacks on European politicians 1990-2004. AB - OBJECTIVE: The only systematic studies of attacks on public figures come from the USA. These studies de-emphasize the role of mental illness and suggest threats are of no predictive value. This study re-examines these questions through a study of attacks on European politicians. METHOD: All non-terrorist attacks on elected politicians in Western Europe between 1990 and 2004 were analysed. RESULTS: Twenty-four attacks were identified, including five involving fatalities, and eight serious injuries. Ten attackers were psychotic, four drunk, nine politically motivated and one unclassifiable. Eleven attackers evidenced warning behaviours. The mentally disordered, most of whom gave warnings, were responsible for most of the fatal and seriously injurious attacks. CONCLUSION: A greater awareness of the link between delusional fixations on public figures and subsequent attacks could aid prevention. Equally importantly, recognition would encourage earlier intervention in people who, irrespective of whether they eventually attack, have delusional preoccupations which ruin their lives. PMID- 17919155 TI - The additional therapeutic effect of group music therapy for schizophrenic patients: a randomized study. AB - OBJECTIVE: Schizophrenia is one of the most serious mental disorders. Music therapy has only recently been introduced as a form of treatment. The aim of this study was to examine the effect of music therapy for schizophrenic in-patients needing acute care. METHOD: Thirty-seven patients with psychotic disorders were randomly assigned to an experimental group and a control group. Both groups received medication and treatment indicated for their disorder. Additionally, the experimental group (n = 21) underwent group music therapy. RESULTS: Significant effects of music therapy are found in patients' self-evaluation of their psychosocial orientation and for negative symptoms. No differences were found in the quality of life. CONCLUSION: Musical activity diminishes negative symptoms and improves interpersonal contact. These positive effects of music therapy could increase the patient's abilities to adapt to the social environment in the community after discharge from the hospital. PMID- 17919157 TI - Height and body mass index in young adulthood and risk of schizophrenia: a longitudinal study of 1 347 520 Swedish men. AB - OBJECTIVE: Measures of body size reflect genetic and environmental influences on growth and energy balance. Associations between such measures and risk of schizophrenia have been inconsistent. METHOD: This is a population-based cohort study of 1 347 520 men born in Sweden from 1952 to 1982, with height and body mass index (BMI) data available from conscription records. The Swedish National Hospital Discharge Register was used to identify subjects diagnosed with schizophrenia from 1970 to 2000. RESULTS: Subjects with lower BMI and shorter height had an increased risk of developing schizophrenia. Underweight subjects had an approximately 30% increase in risk compared with normal BMI subjects (adjusted HR = 1.30, 95% CI: 1.20-1.42). Tall subjects had an approximately 15% reduction in risk compared with short subjects (adjusted HR = 0.85, 95% CI: 0.80 0.92). CONCLUSION: Both height and BMI in early adulthood are strongly and inversely associated with risk of schizophrenia. We discuss these findings in relation to possible genetic and nutritional causal mechanisms. PMID- 17919156 TI - The effects of childhood trauma in patients with first-episode schizophrenia. AB - OBJECTIVE: To evaluate the impact of childhood trauma on psychopathology in 57 patients with first-episode schizophrenia. METHOD: Psychopathology was assessed by Brief Psychiatric Research Scale (BPRS), Scale for the Assessment of Positive Symptoms (SAPS) and Scale for the Assessment of Negative Symptoms (SANS) at first admission. Childhood trauma was assessed by Childhood Abuse Questionnaire and Childhood Trauma Questionnaire (CTQ) after discharge. RESULTS: Frequencies of childhood sexual abuse (CSA), emotional abuse (CEA) and physical abuse (CPA) were reported by 29.8%, 40.9% and 13.6% of the patients respectively. Histories of childhood emotional neglect (CEN) and physical neglect were found in 29.5% and 20.5% of the patients respectively. The patients reporting CSA had higher SAPS scores at admission, and had more suicide attempts before admission. The patients with history of CEA had more hallucinations and delusions of mind reading at admission. CPA, CEA and mean scores of CTQ correlated with the number of siblings. CONCLUSION: Our findings suggest that childhood trauma may alter the presentation of schizophrenia in first admission. PMID- 17919158 TI - Paternal alcoholism predicts the occurrence but not the remission of alcoholic drinking: a 40-year follow-up. AB - OBJECTIVE: To test the effects of father's alcoholism on the development and remission from alcoholic drinking by age 40. METHOD: Subjects were selected from a Danish birth cohort that included 223 sons of alcoholic fathers (high risk; HR) and 106 matched controls (low risk; LR). Clinical examinations were performed at age 40 (n = 202) by a psychiatrist using structured interviews and DSM-III-R diagnostic criteria. RESULTS: HR subjects were significantly more likely than LR subjects to develop alcohol dependence (31% vs. 16%), but not alcohol abuse (17% vs. 15%). More subjects with alcohol abuse were in remission at age 40 than subjects with alcohol dependence. Risk did not predict remission from either alcohol abuse or alcohol dependence. CONCLUSION: Familial influences may play a stronger role in the development of alcoholism than in the remission or recovery from alcoholism. PMID- 17919160 TI - Determinants of coronary heart disease risk for women on a low income: literature review. AB - AIM: This paper is a report of a literature review of the determinants of health and health behaviour relevant to coronary heart disease risk for women living on a low income. BACKGROUND: Coronary heart disease is now recognized as the biggest killer of women in both developed and developing countries. As in men, women's mortality rates for coronary heart disease seem to be directly related to income inequality and social deprivation. METHOD: The Medline, British Nursing Index, CINAHL, Cochrane Library, Psychinfo and Web of Science databases were searched from 1996 to 2006 using the search terms 'women and CHD risk', 'women and health behaviour', 'women low income and health behaviour', 'women low income and smoking', 'women low income and diet' and 'women low income and exercise'. In relation to the wider determinants of health 'women low income and CHD', 'women education and CHD', 'women employment and CHD' and 'women housing and CHD'. Seminal research reports before this period were included if they proved highly influential on later research. A narrative review was conducted. FINDINGS: All the wider determinants of health considered had a negative impact on heart disease risk for women living on low incomes. The latter also appears to have a negative impact on health behaviour. CONCLUSION: Although the impact of the wider determinants of health on coronary heart disease risk are well-understood, their impact on health behaviour (specifically diet, exercise and smoking) is less well understood. If effective interventions are to be designed to tackle inequalities in health, then this deficiency needs to be addressed urgently. PMID- 17919161 TI - Randomized controlled evaluation of a theory-based postpartum sexual health education programme. AB - AIM: This paper reports on a study to examine the effects of a theory-based interactive postpartum sexual health education programme on postpartum women's sexual health knowledge, attitudes towards sexual health, contraceptive self efficacy and sexual self-efficacy. BACKGROUND: Childbearing can challenge marital satisfaction and the sexual life of women, making the childbearing years a vulnerable stage in the sexual life of a woman. Although sexual education used to be a routine aspect of local postpartum teaching, this teaching was inadequate to satisfy women's informational needs related to sexual health. METHODS: For this two-group, randomized controlled trial, 166 participants were recruited at a medical centre in northern Taiwan. The experimental group (n = 84) received the intervention. The control group (n = 82) received routine postpartum teaching. Participants in the experimental group were separated according to their learning preparedness, as determined by the transtheoretical model. Their level of preparedness was then matched to different sexual health education strategies used in the intervention. Data were collected in 2003 at baseline and at 3 days and 8 weeks following the intervention. Descriptive and repeated-measures anova were used to analyse data. RESULTS: Sexual health knowledge, attitudes and sexual self-efficacy were significantly greater for women in the experimental group at 3 days and 8 weeks after the intervention programme than for those in the control group. However, contraceptive self-efficacy was not significantly different in the two groups. CONCLUSION: The longer-term effectiveness of our theory-based interactive postpartum sexual health education programme will be enhanced by matching teaching strategies to participants' stage of learning preparedness. PMID- 17919162 TI - Successful blind placement of nasojejunal tubes in paediatric intensive care: impact of training and audit. AB - AIM: This paper is a report of a study to monitor continuing nasojejunal tube placement success rate and to evaluate a training programme for staff placing these tubes at the bedside. BACKGROUND: Gastric delivery of enteral feeds is frequently poorly tolerated due to impaired gastric motility in critically ill children. Consequently, there has been an increased interest in the use of the nasojejunal feeding route. Nasojejunal tubes are both safe and well-tolerated, but placements of these tubes are notoriously difficult and therefore often avoided. Consequently, a blind bedside technique was developed, with a placement success of 96%. METHOD: A training programme using this technique was developed for nursing staff on the unit. This included one-to-one training of key nurses of the nasojejunal tube placement technique and a supervised nasojejunal placement with either the senior nurse or dietitian. This practice was audited in 100 consecutive patients in 2001 and 2004. Patient demographics, diagnosis, time taken for placement and reasons for unsuccessful placements were documented. RESULTS: The 2001 audit (n = 100) indicated that the nasojejunal route was used in 19% of all cases, with 1% and 80% of patients fed via the parenteral and nasogastric route respectively. In 2004 (n = 94), 18% of patients were fed via the nasojejunal route, 3% parenterally and 79% via the gastric route. Placement success continued to be between 94.5-95% in 2001 and 2004. CONCLUSION: Continued successful placement of blind nasojejunal feeding tubes can be achieved through a well-monitored education programme, regular audit cycles and multidisciplinary team support. PMID- 17919163 TI - The impact of team processes on psychiatric case management. AB - AIM: This paper is a report of a study to identify the structures and interactions within community mental health teams that facilitate or impede effective teamwork and psychiatric case management. BACKGROUND: Effective case management requires close collaboration between case managers or care co ordinators and other members of the multidisciplinary mental health team, yet there has been little research into this relationship. METHOD: A multiple case study of seven United Kingdom community mental health teams was conducted between 1999 and 2001, using qualitative methods of participant observation, semi structured interviews and document review. FINDINGS: Factors were identified that impacted on the ability of care co-ordinators to act effectively: 'structure and procedures'; 'disrespect and withdrawal'; 'humour and undermining'; 'safety and disclosure'. Care co-ordination was enhanced when team structures and policies were in place and where team interactions were respectful. Where members felt disrespected or undermined, communication, information sharing and collaboration were impaired, with a negative impact on the care provided to service users. CONCLUSIONS: Teams require clear operating procedures alongside interprofessional trust and respect to ensure that there is open, safe and reflective participation. Further research is required to identify how best to bring about collaborative, effective teamwork in mental healthcare. PMID- 17919164 TI - Evaluation of three fall-risk assessment tools in an acute care setting. AB - AIM: This paper is a report of a study to evaluate the validity of three fall risk assessment tools to identify patients at high risk for falls. BACKGROUND: Patient falls make up 38% of all adverse events occurring in hospital settings, and may result in physical injury and undesirable emotional and financial outcomes. No single fall-risk assessment tool has been conclusively validated. METHOD: The Morse Fall Scale, St Thomas Risk Assessment Tool in Falling Elderly Inpatients, and Hendrich II Fall Risk Model were validated in inter-rater reliability and validity studies in 2003. This included assessment of the probability of disagreement, kappa-values, sensitivity, specificity, positive predictive values and negative predictive values of the assessment tools with the associated 95% CI. FINDINGS: One hundred and forty-four patients were recruited for the inter-rater reliability study. The probabilities of disagreement were between 2.8% and 9.7%, and 95% CI for all tools ranged from 1.1% to 15.7%. The kappa-values were all higher than 0.80. In the validity study, 5489 patients were recruited to observe 60 falls. The Morse Fall Scale at a cutoff score of 25 and Heindrich II Fall Risk Model at a cutoff score of 5 had strong sensitivity values of 88% and 70%, respectively. However, in comparison with the Morse Fall Scale (specificity = 48.3%), only the Heindrich II Fall Risk Model had a more acceptable level of specificity (61.5%). CONCLUSION: The Heindrich II Fall Risk Model is potentially useful in identifying patients at high risk for falls in acute care facilities. PMID- 17919172 TI - Of mighty mice and diabetic men: animal models of islet dysfunction. PMID- 17919173 TI - Caveats and considerations for performing pancreas-specific gene manipulations in the mouse. AB - Conditional gene targeting using the Cre/loxP strategy has proven to be very useful for studies of glucose homeostasis, tissue function and dysfunction in diabetes, and pancreas development. However, use of this strategy over the past decade has revealed a variety of experimental caveats, many of which are a direct consequence of the procedures used to generate Cre-driver lines. We discuss frequently encountered experimental artefacts, the advantages of using bacterial artificial chromosome-derived transgenes or performing a Cre knockin for improving the specificity of expression, and systems for regulating Cre activity. In addition, recent studies indicate that high amounts of Cre in the pancreatic beta-cell may cause glucose intolerance and impaired insulin secretion. However, these findings, while serving as a reminder for simple experimental controls, are unlikely to diminish utilization of this very powerful and useful technology. PMID- 17919174 TI - Unexpected functional consequences of xenogeneic transgene expression in beta cells of NOD mice. AB - We describe unexpected alterations in the non-obese diabetic (NOD/Lt) mouse model of type 1 diabetes (T1D) following forced beta-cell expression of non-mammalian genes ligated to an insulin promoter sequence. These include the jellyfish green fluorescent protein (GFP), useful for beta-cell identification, and the bacteriophage P1 Cre recombinase, necessary for beta cell-specific ablation of a gene using a Cre-loxP system. Homozygous expression of GFP, driven by the mouse insulin 1 gene promoter (MIP-GFP) in a single transgenic line of NOD mice, produced T1D in postnatal mice that was not associated with insulitis, but rather beta cell-depleted islets. Hemizygous transgene expression suppressed spontaneous autoimmune T1D in females, and produced a male glucose intolerance syndrome associated with age-dependent declines in plasma insulin content. Among lines of transgenic NOD/Lt mice expressing Cre recombinase driven by the rat insulin 2 promoter (RIP-Cre), high, non-mosaic expression correlated with suppressed T1D development. These findings emphasize the need for careful characterization of genetically manipulated NOD mouse stocks to insure that model characteristics have not been compromised. PMID- 17919175 TI - Sirtuins and beta-cell function. AB - Over the past century, there have been major advances in medicine, diet and living conditions that have substantially extended the health span of people in the developed world. Over the past decade or so, the science of ageing has made great strides in providing an understanding of the underlying causes of ageing and how diet and genes play into the ageing process. In this essay, we will review some of the recent advances made in the science of sirtuins and beta-cell biology, and discuss how we will apply this knowledge to further the progress of humankind in healthy ageing. PMID- 17919176 TI - Mutations in the ABCC8 gene encoding the SUR1 subunit of the KATP channel cause transient neonatal diabetes, permanent neonatal diabetes or permanent diabetes diagnosed outside the neonatal period. AB - AIM: Mutations in the ABCC8 gene encoding the SUR1 subunit of the pancreatic ATP sensitive potassium channel cause permanent neonatal diabetes mellitus (PNDM) and transient neonatal diabetes mellitus (TNDM). We reviewed the existing literature, extended the number of cases and explored genotype-phenotype correlations. METHODS: Mutations were identified by sequencing in patients diagnosed with diabetes before 6 months without a KCNJ11 mutation. RESULTS: We identified ABCC8 mutations in an additional nine probands (including five novel mutations L135P, R306H, R1314H, L438F and M1290V), bringing the total of reported families to 48. Both dominant and recessive mutations were observed with recessive inheritance more common in PNDM than TNDM (9 vs. 1; p < 0.01). The remainder of the PNDM probands (n = 12) had de novo mutations. Seventeen of twenty-five children with TNDM inherited their heterozygous mutation from a parent. Nine of these parents had permanent diabetes (median age at diagnosis: 27.5 years, range: 13-35 years). Recurrent mutations of residues R1183 and R1380 were found only in TNDM probands and dominant mutations causing PNDM clustered within exons 2-5. CONCLUSIONS: ABCC8 mutations cause PNDM, TNDM or permanent diabetes diagnosed outside the neonatal period. There is some evidence that the location of the mutation is correlated with the clinical phenotype. PMID- 17919177 TI - The HNF-1alpha-SNARE connection. AB - Maturity-onset diabetes of the young (MODY) is a monogenic form of type 2 diabetes mellitus that is characterized by impairment of glucose-stimulated insulin secretion from pancreatic beta-cells. We previously reported that heterozygous mutations of the hepatocyte nuclear factor (HNF)-1alpha gene cause a form of MODY (MODY3). We have subsequently found that collectrin, a recently cloned kidney-specific gene of unknown function, is a novel target of HNF-1alpha in pancreatic beta-cells. In addition, we have demonstrated that collectrin forms a complex with the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex by direct interaction with snapin, a protein that is thought to be involved in neurotransmission by binding to synaptosomal-associated protein, 25 KD (SNAP25). Collectrin favours the formation of SNARE complexes and controls insulin exocytosis. PMID- 17919178 TI - Functional analysis of two Kir6.2 (KCNJ11) mutations, K170T and E322K, causing neonatal diabetes. AB - Heterozygous activating mutations in Kir6.2 (KCNJ11), the pore-forming subunit of the adenosine triphosphate (ATP)-sensitive potassium (K(ATP)) channel, are a common cause of neonatal diabetes (ND). We assessed the functional effects of two Kir6.2 mutations associated with ND: K170T and E322K. K(ATP) channels were expressed in Xenopus oocytes, and the heterozygous state was simulated by coexpression of wild-type and mutant Kir6.2 with SUR1 (the beta cell type of sulphonylurea receptor (SUR)). Both mutations reduced the sensitivity of the K(ATP) channel to inhibition by MgATP and enhanced whole-cell K(ATP) currents. In pancreatic beta cells, such an increase in the K(ATP) current is expected to reduce insulin secretion and thereby cause diabetes. The E322K mutation was without effect when Kir6.2 was expressed in the absence of SUR1, suggesting that this residue impairs coupling to SUR1. This is consistent with its predicted location on the outer surface of the tetrameric Kir6.2 pore. The kinetics of K170T channel opening and closing were altered by the mutation, which may contribute to the lower ATP sensitivity. Neither mutation affected the sensitivity of the channel to inhibition by the sulphonylurea tolbutamide, suggesting that patients carrying these mutations may respond to these drugs. PMID- 17919179 TI - The balance between proinsulin biosynthesis and insulin secretion: where can imbalance lead? AB - Insulin is stored in pancreatic beta-cells in beta-granules. Whenever insulin is secreted in response to a nutrient secretagogue, there is a complementary increase in proinsulin biosynthesis to replenish intracellular insulin stores. This specific nutrient regulation of proinsulin biosynthesis is predominately regulated at the translational level. Recently, a highly conserved cis-element in the 5'-untranslated region (UTR) of preproinsulin mRNA, named ppIGE, has been identified that is required for specific translational regulation of proinsulin biosynthesis. This ppIGE is also found in the 5'-UTR of certain other translationally regulated beta-granule protein mRNAs, including the proinsulin processing endopeptidases, PC1/3 and PC2. This provides a mechanism whereby proinsulin processing is adaptable to changes in proinsulin biosynthesis. However, relatively few beta-granules undergo secretion, with most remaining in the storage pool for approximately 5 days. Aged beta-granules are retired by intracellular degradation mechanisms, either via crinophagy and/or autophagy, as another long-term means of maintaining beta-granule stores at optimal levels. When a disconnection between insulin production and secretion arises, as may occur in type 2 diabetes, autophagy further increases to maintain beta-granule numbers. However, if this increased autophagy becomes chronic, autophagia mediated cell death occurs that could then contribute to beta-cell loss in type 2 diabetes. PMID- 17919180 TI - microRNAs and the regulation of glucose and lipid metabolism. AB - Over recent years, metabolic disorders such as type 2 diabetes have finally become recognized as a major challenge to global health. The attention of scientists therefore has to focus on improving our understanding of the molecular mechanisms behind these diseases and towards the design of new drug therapy strategies. The pathophysiology of diabetes is undoubtedly complex, oftentimes characterized by varying states of insulin resistance and impaired beta-cell function; however, the identification of new pathways is constantly improving our understanding of the disease. We and others have recently shown that microRNAs (miRNAs) can play a role in insulin secretion and glucose homostasis. Thus, in this review, we will discuss the potential role of miRNAs in type 2 diabetes and related metabolic diseases. PMID- 17919181 TI - A transgenic model to study the pathogenesis of somatic mtDNA mutation accumulation in beta-cells. AB - Low levels of somatic mutations accumulate in mitochondrial DNA (mtDNA) as we age; however, the pathogenic nature of these mutations is unknown. In contrast, mutational loads of >30% of mtDNA are associated with electron transport chain defects that result in mitochondrial diseases such as mitochondrial encephalopathy lactic acidosis and stroke-like episodes. Pancreatic beta-cells may be extremely sensitive to the accumulation of mtDNA mutations, as insulin secretion requires the mitochondrial oxidation of glucose to CO(2). Type 2 diabetes arises when beta-cells fail to compensate for the increased demand for insulin, and many type 2 diabetics progress to insulin dependence because of a loss of beta-cell function or beta-cell death. This loss of beta-cell function/beta-cell death has been attributed to the toxic effects of elevated levels of lipids and glucose resulting in the enhanced production of free radicals in beta-cells. mtDNA, localized in close proximity to one of the major cellular sites of free radical production, comprises more than 95% coding sequences such that mutations result in changes in the coding sequence. It has long been known that mtDNA mutations accumulate with age; however, only recently have studies examined the influence of somatic mtDNA mutation accumulation on disease pathogenesis. This article will focus on the effects of low-level somatic mtDNA mutation accumulation on ageing, cardiovascular disease and diabetes. PMID- 17919182 TI - beta-cell hyperexcitability: from hyperinsulinism to diabetes. AB - Nutrient oxidation in beta cells generates a rise in [ATP]:[ADP] ratio. This reduces K(ATP) channel activity, leading to depolarization, activation of voltage dependent Ca(2+) channels, Ca(2+) entry and insulin secretion. Consistent with this paradigm, loss-of-function mutations in the genes (KCNJ11 and ABCC8) that encode the two subunits (Kir6.2 and SUR1, respectively) of the ATP-sensitive K(+) (K(ATP)) channel underlie hyperinsulinism in humans, a genetic disorder characterized by dysregulated insulin secretion. In mice with genetic suppression of K(ATP) channel subunit expression, partial loss of K(ATP) channel conductance also causes hypersecretion, but unexpectedly, complete loss results in an undersecreting, mildly glucose-intolerant phenotype. When challenged by a high fat diet, normal mice and mice with reduced K(ATP) channel density respond with hypersecretion, but mice with more significant or complete loss of K(ATP) channels cross over, or progress further, to an undersecreting, diabetic phenotype. It is our contention that in mice, and perhaps in humans, there is an inverse U-shaped response to hyperexcitabilty, leading first to hypersecretion but with further exacerbation to undersecretion and diabetes. The causes of the overcompensation and diabetic susceptibility are poorly understood but may have broader implications for the progression of hyperinsulinism and type 2 diabetes in humans. PMID- 17919184 TI - New insights into the molecular mechanisms of priming of insulin exocytosis. AB - Exocytosis of insulin vesicles in the pancreatic beta-cell involves a sequence of regulated events, whose normal function and efficient adaptation to increased demand are essential for the maintenance of glucose homeostasis. These exocytotic events comprise the trafficking and docking of vesicles to the plasma membrane, followed by fusion triggered by Ca(2+). Recent studies have unravelled post docking steps mediated by novel factors, which, by their interactions with soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)- and SNARE-associated proteins, confer the docked vesicles fusion competence. These priming steps define the releasable pool of insulin vesicles, which accounts for the first phase of insulin secretion, and controls the rate at which vesicles are replenished for the second phase of secretion. This article aims to summarize what is currently known about the mechanisms that underlie the priming activity of these proteins, focusing on Munc13, a topic to which we have made some recent contributions. Abnormal glucose homeostasis in type 2 diabetes is because of the failure of islet beta-cells to augment insulin secretion sufficiently to compensate for reduced insulin sensitivity. A better understanding of the priming steps may help develop novel approaches to increase insulin secretory capacity and thereby prevent the progression to type 2 diabetes. PMID- 17919183 TI - Action potentials and insulin secretion: new insights into the role of Kv channels. AB - Coordinated electrical activity allows pancreatic beta-cells to respond to secretagogues with calcium entry followed by insulin secretion. Metabolism of glucose affects multiple membrane proteins including ion channels, transporters and pumps that collaborate in a cascade of electrical activity resulting in insulin release. Glucose induces beta-cell depolarization resulting in the firing of action potentials (APs), which are the primary electrical signal of the beta cell. They are shaped by orchestrated activation of ion channels. Here we give an overview of the voltage-gated potassium (Kv) channels of the beta-cell, which are responsible in part for the falling phase of the AP, and how their regulation affects insulin secretion. beta cells contain several Kv channels allowing dynamic integration of multiple signals on repolarization of glucose-stimulated APs. Recent studies on Kv channel regulation by cAMP and arachidonic acid and on the Kv2.1 null mouse have greatly increased our understanding of beta-cell excitation-secretion coupling. PMID- 17919185 TI - Secretory vesicle docking to the plasma membrane: molecular mechanism and functional significance. AB - In regulated exocytic pathways, secretion occurs only in the presence of appropriate stimuli. Professional secretory cells harbour specific storage organelles that release bioactive substances with both controlled timing and quantity in response to the strength and period of stimulation. Although each secretory organelle is highly differentiated in multicellular organisms, the basic regulatory mechanism is thought to be conserved. In most instances, the secretagogue increases the intracellular Ca(2+) concentration from the resting level of approximately 100 nM to somewhere between approximately 10 and 100 microM. Although Ca(2+) sensors of the final fusion reaction, such as synaptotagmin, have been investigated intensively in synaptic vesicle exocytosis, there are other preceding rate-limiting steps influenced by Ca(2+) and other secretory signals, especially in the exocytosis of secretory granules whose time course is much slower than that of synaptic vesicles. The stable docking of secretory vesicles to the fusion site that is only seen in regulated exocytic pathways may represent one such critical step. Here, we review the molecular mechanism of docking, mainly based on recent findings on insulin granules in pancreatic beta cells, and propose a new concept for its functional significance in regulated exocytosis. PMID- 17919186 TI - Islet-cell-to-cell communication as basis for normal insulin secretion. AB - The emergence of pancreatic islets has necessitated the development of a signalling system for the intra- and inter-islet coordination of beta cells. With evolution, this system has evolved into a complex regulatory network of partially cross-talking pathways, whereby individual cells sense the state of activity of their neighbours and, accordingly, regulate their own level of functioning. A consistent feature of this network in vertebrates is the expression of connexin (Cx)-36-made cell-to-cell channels, which cluster at gap junction domains of the cell membrane, and which adjacent beta cells use to share cytoplasmic ions and small metabolites within individual islets. This chapter reviews what is known about Cx36, and the mechanism whereby this beta-cell connexin significantly regulates insulin secretion. It further outlines other less established functions of the protein and evaluates its potential relevance for the development of novel therapeutic approaches to diabetes. PMID- 17919187 TI - Sterol regulatory element-binding protein-1c and pancreatic beta-cell dysfunction. AB - It has long been known that excess intracellular fatty acids cause impaired insulin secretion, referred to as beta-cell lipotoxicity. Sterol regulatory element-binding protein (SREBP)-1c is a transcription factor that controls hepatic fatty acid synthesis. Activation of SREBP-1c by overnutrition also inhibits insulin receptor substrate-2 (IRS-2) and induces insulin resistance in the liver. As SREBP-1c is also expressed in beta cells, we hypothesized that activation of SREBP-1c could be a part of the mechanism by which saturated fatty acids induce beta-cell lipotoxicity. We found that nuclear SREBP-1c has a negative impact on both glucose- and potassium-stimulated insulin secretion as determined in islets from beta-cell-specific SREBP-1c transgenic mice as well as SREBP-1c knockout mice. This effect of SREBP-1c involves multiple functional pathways required for insulin secretion from beta cells: (i) decreased ATP caused by energy consumption through lipogenesis and uncoupling protein-2 (UCP-2) activation; (ii) repressed IRS-2 and pancreas duodenum homeobox 1 (PDX1) expression, leading to impaired beta-cell mass; and (iii) impaired post-ATP membrane voltage-dependent steps of the insulin secretion pathway caused by upregulated granuphilin and other ion channel proteins. Saturated fatty acids, such as palmitic acid (PA), impair insulin secretion through SREBP-1c activation, whereas polyunsaturated fatty acids including eicosapentaenoic acid (EPA) restore PA-suppressed insulin secretion through suppression of SREBP-1c. These data implicate a therapeutic potential of EPA against insulin secretion defects caused by lipotoxicity. PMID- 17919188 TI - Regulation of pancreatic beta-cell function by the forkhead protein FoxO1. AB - Forkhead transcription factors of the FoxO family play a critical role in cellular differentiation, proliferation, apoptosis and stress resistance. FoxO1 regulates glucose and lipid production in liver; food intake in the hypothalamus and cell differentiation in preadipocytes, myoblasts and vascular endothelium. In this review, we summarize recent literature on the role of FoxO1 in pancreatic beta cells. FoxO1 regulates beta-cell proliferation and protects against beta cell failure induced by oxidative stress through NeuroD and MafA induction. In addition, FoxO1 nuclear exclusion is required for the proliferative effects of glucoincretin glucagon-like peptide-1 in islets. The data begin to outline an overarching role of FoxO1 in beta-cell function. PMID- 17919189 TI - Regulation of beta-cell mass and function by the Akt/protein kinase B signalling pathway. AB - The insulin receptor substrate-2/phosphoinositide 3-kinase (PI3K) pathway plays a critical role in the regulation of beta-cell mass and function, demonstrated both in vitro and in vivo. The serine threonine kinase Akt is one of the promising downstream molecules of this pathway that has been identified as a potential target to regulate function and induce proliferation and survival of beta cells. Here we summarize some of the molecular mechanisms, downstream signalling pathways and critical components involved in the regulation of beta-cell mass and function by Akt. PMID- 17919190 TI - Role of the M3 muscarinic acetylcholine receptor in beta-cell function and glucose homeostasis. AB - The release of insufficient amounts of insulin in the presence of elevated blood glucose levels is one of the key features of type 2 diabetes. Various lines of evidence indicate that acetylcholine (ACh), the major neurotransmitter of the parasympathetic nervous system, can enhance glucose-stimulated insulin secretion from pancreatic beta-cells. Studies with isolated islets prepared from whole body M(3) muscarinic ACh receptor knockout mice showed that cholinergic amplification of glucose-dependent insulin secretion is exclusively mediated by the M(3) muscarinic receptor subtype. To investigate the physiological relevance of this muscarinic pathway, we used Cre/loxP technology to generate mutant mice that lack M(3) receptors only in pancreatic beta-cells. These mutant mice displayed impaired glucose tolerance and significantly reduced insulin secretion. In contrast, transgenic mice overexpressing M(3) receptors in pancreatic beta-cells showed a pronounced increase in glucose tolerance and insulin secretion and were resistant to diet-induced glucose intolerance and hyperglycaemia. These findings indicate that beta-cell M(3) muscarinic receptors are essential for maintaining proper insulin secretion and glucose homeostasis. Moreover, our data suggest that enhancing signalling through beta-cell M(3) muscarinic receptors may represent a new avenue in the treatment of glucose intolerance and type 2 diabetes. PMID- 17919191 TI - Evaluation of beta-cell mass and function in the Gottingen minipig. AB - Increased knowledge about beta-cell mass and function is important for our understanding of the pathophysiology of type 2 diabetes (T2DM). The relationship between the two is difficult to study in humans, whereas animal models allow studies of consequences of, for example, reduction of beta-cell mass and induction of obesity and procurement of the pancreas for histological examination. An overview of results obtained in the Gottingen minipig in relation to beta-cell function, and mass is provided here. Effects of a primary reduction of beta-cell mass have indicated that not all of the defects of pulsatile insulin secretion in human T2DM can be explained by reduced beta-cell mass. Furthermore, induction of obesity has shown deterioration of beta-cell function and morphological changes in the pancreas. As in humans, obesity leads to an increased beta-cell volume in the minipig, and based on the increased number of islets, neogenesis of islets is an important factor in expansion of beta-cell mass in this species. Measurement of beta-cell function as an estimate of beta cell mass is, at present, the only method possible in humans, and this approach has been validated using lean and obese minipigs with a range of beta-cell mass. The effects on beta-cell function and mass of obesity of longer duration and/or more pronounced hyperglycaemia remains to be determined, but the models developed so far represent a valuable tool for such investigations. PMID- 17919192 TI - Islet gene expression and function in type 2 diabetes; studies in the Goto Kakizaki rat and humans. AB - Defective beta-cell function with resulting impairment of glucose-stimulated insulin release is a critical factor in the pathogenesis of type 2 diabetes. Accumulated studies in pancreatic islets of the spontaneously diabetic Goto Kakizaki (GK) rat suggest that this is a useful animal model of type 2 diabetes. The GK rat is non-obese, and abnormal glucose regulation develops early in life in association with impaired insulin secretion. There are some differences in islet morphology and function reported between different GK rat colonies. In addition to reduction of beta-cell mass, a number of beta-cell defects have been described with possible relevance for the reduced insulin secretion. Interestingly, some of these defects have also been shown in isolated islets from type 2 diabetic humans. The polygenic nature of diabetes heredity in the GK rat may well resemble the genetic basis in the majority of patients with type 2 diabetes. Here, we review studies concerning beta-cell function and islet gene expression in the GK rat and compare it with the limited number of investigations on similar topics in isolated islets from patients with type 2 diabetes. PMID- 17919193 TI - Type 2 diabetes - a matter of failing beta-cell neogenesis? Clues from the GK rat model. AB - Now that reduction in beta-cell mass has been clearly established in humans with type 2 diabetes mellitus (T2D), the debate focuses on the possible mechanisms responsible for decreased beta-cell number. Appropriate inbred rodent models are essential tools for this purpose. The information available from the Goto Kakizaki (GK) rat, one of the best characterized animal models of spontaneous T2D, is reviewed in such a perspective. We propose that the defective beta-cell mass in the GK model reflects mostly a persistently decreased beta-cell neogenesis. The data discussed in this review are consistent with the notion that poor proliferation and/or survival of the endocrine precursor cells during GK foetal life will result in a decreased pool of endocrine precursors in the pancreas, and hence an impaired capacity of beta-cell neogenesis (either primary in the foetus or compensatory in the newborn and the adult). As we also demonstrated that beta-cell neogenesis can be pharmacologically reactivated in the GK model, our work supports, on a more prospective basis, the concept that facilitation of T2D treatment may be obtained through beta-cell mass expansion after stimulation of beta-cell regeneration/neogenesis in diabetic patients. PMID- 17919194 TI - Intrauterine programming of the endocrine pancreas. AB - Epidemiological studies have revealed strong relationships between poor foetal growth and subsequent development of the metabolic syndrome. Persisting effects of early malnutrition become translated into pathology, thereby determine chronic risk for developing glucose intolerance and diabetes. These epidemiological observations identify the phenomena of foetal programming without explaining the underlying mechanisms that establish the causal link. Animal models have been established and studies have demonstrated that reduction in the availability of nutrients during foetal development programs the endocrine pancreas and insulin sensitive tissues. Whatever the type of foetal malnutrition, whether there are not enough calories or protein in food or after placental deficiency, malnourished pups are born with a defect in their beta-cell population that will never completely recover, and insulin-sensitive tissues will be definitively altered. Despite the similar endpoint, different cellular and physiological mechanisms are proposed. Hormones operative during foetal life like insulin itself, insulin-like growth factors and glucocorticoids, as well as specific molecules like taurine, or islet vascularization were implicated as possible factors amplifying the defect. The molecular mechanisms responsible for intrauterine programming of the beta cells are still elusive, but two hypotheses recently emerged: the first one implies programming of mitochondria and the second, epigenetic regulation. PMID- 17919195 TI - Proceedings of the L'Oreal Workshop on African Hair and Skin, November 2-3, 2006, Dakar, Senegal. PMID- 17919196 TI - Worldwide diversity of hair curliness: a new method of assessment. AB - BACKGROUND: For many years, cosmetic scientists have attempted to measure the physical features of human hair, such as its shape and colour, as these can be artificially modified using cosmetic products. With regard to hair shape, previous anthropologic studies have emphasized its variability within and between human ethnic groups. Many studies have broadly distinguished three ethnic human subgroups: African, Asian, and Caucasian. Such a broad classification cannot account for the great complexity of human biological diversity, resulting from multiple, past or recent mixed origins. The verbal description of hair shape ranges from the classic to the more sophisticated, with terms such as straight, wavy, curly, frizzy, kinky, woolly, helical, etc. Although these descriptions evoke a global appearance, they remain confusing as their definitions and limits are unclear. Assessments are therefore required to more accurately define such verbal attributes. OBJECTIVE: The work reported here attempts to address the following issues: (i) to define hair types according to specific shape criteria through objective and simple measurements; and (ii) to define such hair types without referring to human ethnicity. METHODS: Measurements of four parameters related to hair curliness (curve diameter, curl index) or kinking of the hair (numbers of waves, numbers of twists) were performed on hair from 2449 subjects from 22 different countries. Principal components analysis and hierarchical ascendant classification were used to identify homogeneous groups of hair and to determine key variables for the assignment of group membership. Finally, a segmentation tree was prepared in order to establish simple rules for predicting group membership of new subjects. RESULTS AND CONCLUSIONS: This study has shown that it is possible to classify the various hair types found worldwide into eight main groups. The approach involves objective descriptors of hair shape, and is more reliable than traditional methods relying on categories such as curly, wavy, and kinky. Applied to worldwide human diversity, it avoids reference to the putative, unclear ethnic origin of subjects. Briefly, a straight hair type I is just that, and whether it originates from a Caucasian or an Asian subject is not at issue. The hair types defined here also more adequately reflect the large variation of hair shape diversity around the world, and may possibly help to trace past mixed origins amongst human subgroups. PMID- 17919198 TI - Comparative study of the hydration of the stratum corneum between four ethnic groups: influence of age. AB - BACKGROUND: Several recent overviews have reported that significant work remains to be performed to understand and quantify the ethnic differences in skin properties. In this way, we have carried out a set of in vivo biophysical experiments on the skin of American women from different ethnic populations living in the same environment. Inter- and intraethnic skin micro relief results were already published, skin hydration differences are now reported here. METHODS: The skin water content was evaluated taking advantage of the new skin capacitance imaging technique which allows to study stratum corneum hydration without suffering of the influence of the skin micro-relief and hair on the measurement. Three hundred and eleven American women from four ethnic groups were enrolled in this study. The investigation was performed during the summer season of 2004 on the major relatively distinct ethnic groups of Chicago, which is to say: African American, Chinese, Caucasian and Mexican. The hydration of the skin was investigated on the dorsal and ventral forearm sites as a function of ethnicity and age. RESULTS: Skin dryness is higher on sun exposed sites for lighter skin tones, such as in Chinese and Caucasian women, than on sites that are primarily out of the sun; while, no skin dryness differences are seen on either site for African American and Mexican women whose skin is darker. The skin dryness does not change as a function of ethnicity for the younger group for either the ventral and dorsal site of the forearm. With age, however, the dryness of the skin is higher for African American and Caucasian women than for the two other ethnic groups, with a higher percentage increase in Caucasian women. CONCLUSIONS: This study has revealed that the hydration of the skin is different according to ethnicity and that the age effects are influenced by ethnicity, suggesting anatomical or physiological property differences in ethnic skin. This study has also pointed out that the SkinChip seems to be a convenient and fast way to investigate both the micro relief as previously published and the dryness of the skin on a large number of subjects, and in this way will be very useful to improve our knowledge about skin of people from different ethnic groups and helping to develop specific products that are customized to all these populations. PMID- 17919197 TI - Human hair keratin network and curvature. AB - BACKGROUND: In human hair, very little is known about the substructures relating to the curl pattern. The interpretation of the macroscopic shape of the fiber at the molecular and cellular scales is still unclear. METHODS: A comparative and multiscale study was carried out on a set of human hair samples, ranging in shape from straight to tightly curled, in order to investigate structural elements that might be related to hair curl pattern. RESULTS: At the macroscopic level, the frequency and amplitude of cross-sectional rotations were found to be crucial for an accurate description of curliness. At the cellular level, transmission electron microscopy experiments made it possible to confirm that macrofibril organization was strongly related to hair curliness. The curly hair follicles exhibited retrocurvature, independent of ethnic origin. A direct comparison of straight hair and curly hair highlighted an intrinsic asymmetry in the proliferative compartment that clearly extended above the Auber line on the convex side of the curvature. This phenomenon caused delayed differentiation of both inner and outer root sheaths. The hair cortex itself was elliptical and asymmetric, as evidenced by hHa8 keratin. In curly hair, this hair keratin accumulated on the concave side of the curvature, whereas, in straight hair, positive cortical cells were evenly distributed throughout the circular fiber. CONCLUSION: The curly shape of the hair shaft seems to result from the asymmetric differentiation of the precortex. Hair fiber can thus be considered as a shape memory material. PMID- 17919200 TI - Hair loss in children in South-East Nigeria: common and uncommon cases. AB - BACKGROUND: The cosmetic importance of hair as a unique structure of the scalp is immense, such that once the loss of hair becomes an issue for a child, parents, in most cases, project their own concerns, often expressed as guilt. The clinical presentation of hair loss and scalp disorders in children varies widely, often ranging from subtle to disfiguring forms, and may be acquired or congenital. This variable pattern of hair loss is seen in all regions, possibly based on the ethnic origin of hair types. AIM: To determine the clinical aspects of hair loss based on clinical appearance, age of onset, duration of disease, and associated features, and to provide a guide for the evaluation of acquired hair loss and scalp disorders commonly occurring in childhood in south-east Nigeria. METHODS: One hundred and thirteen children with hair loss and scalp disorders consulted the University of Nigeria Teaching Hospital Skin Clinic between February 2002 and March 2003. The data collected included age, sex, clinical presentation, associated symptoms, and family history. Laboratory tests [full blood count (FBC), thyroid function test (TFT), antinuclear antibody (ANA), urinalysis], microscopic examination, Wood's light examination, and histology were conducted for further confirmation where necessary. RESULTS: There were 71 males (62.8%) and 42 females (37.2%), aged between 3 months and 13 years. The presenting symptoms included scaly scalp, itchiness, and anxiety; some patients were asymptomatic. Hair loss and fear were the chief concerns of 89.6% and 51.3% of parents, respectively, who presented with their children. Commonly occurring diagnoses were tinea capitis (62; 54.9%), alopecia areata (43; 38.1%), psoriasis (5; 4.4%), and telogen effluvium (3; 2.7%). Seventeen patients with alopecia areata had atopic dermatitis and three had vitiligo. CONCLUSION: Commonly occurring hair loss in children in our region is mainly acquired, and the clinical course is related to the parent's attitude to treatment, particularly for tinea capitis. PMID- 17919199 TI - Aesthetic problems associated with the cosmetic use of bleaching products. AB - BACKGROUND AND AIM: The use of skin bleaching products for cosmetic purposes is a frequent practice (25-96%) in women from sub-Saharan Africa. The dermatologic complications associated with this practice have been comprehensively reported. The aim of this work was to study the epidemiologic, clinical, and cosmetic aspects of these complications in order to produce better therapeutic guidelines for their management. METHODS: This was a prospective, descriptive study performed over a 6-month period. All women aged between 15 and 50 years, who consulted a dermatologist (Le Dantec Hospital or Institute of Social Hygiene), experienced a complication associated with artificial depigmentation, and agreed to take part in the study, were included. The data were input and analyzed using Epi info version 6.0. RESULTS: Eighty-six female patients were included, with a mean age of 29.34 years (range, 16-49 years). The breakdown by level of education was as follows: primary (48.8%), secondary (18.3%), and higher (8.5%) education. Twenty-two per cent of our population had not attended school. The mean monthly cost of skin bleaching products was 6.22 euros. The initial skin tone before using skin bleaching products was black in 41.5% of patients, light in 32.9%, and intermediate in 25.6%. The mean duration of exposure was 6.7 +/- 5 years (range, 1-30 years). The breakdown by skin bleaching products showed that topical corticosteroids were the most frequently used (78%), followed by hydroquinone (56%), products based on vegetable extracts (31.7%), caustic products (8.5%), and, finally, products of unknown composition (41.4%). Two components or more were frequently combined (86.5%). The aesthetic complications of artificial depigmentation were the reason for consulting a dermatologist in 10 patients (12%). Nineteen types of aesthetic complication were reported in our sample. Hyperpigmentation of the joints was the most frequently found complication (85.4%), followed by striae atrophicae (72%) and skin atrophy (59.8%). The number of aesthetic complications found in the patients varied from one to nine. Patients frequently presented (71.9%) with other complications associated with artificial depigmentation. CONCLUSIONS: Aesthetic complications associated with artificial depigmentation are common, but rarely the reason for consulting a dermatologist. In the absence of suitable therapeutic agents, prevention, based on informing women of the damaging effects of artificial depigmentation, is the only way forward. PMID- 17919201 TI - Impact of hair relaxers in women in Nakuru, Kenya. AB - BACKGROUND: The practice of relaxing hair is intended to straighten it, so that it is easy to manage and style, and the result is aesthetically pleasing. We studied the impact of using hair relaxers in women in Nakuru, Kenya. METHODS: A cross-sectional study was carried out between February and March 2006. The subjects were women and girls aged 15 years and above, living in Nakuru district, Kenya, who had been or were currently using hair relaxers and were willing to participate. Sampling was by the snowball method and data collection by questionnaire. The data were analyzed using Epi-info 2000. RESULTS: Two hundred and forty-two women were recruited into the study. The age range was 15-51 years, with a median of 32 years. The main reasons given for relaxing hair were that it made hair easy to manage and looked "beautiful". One hundred and thirty-four women (67%) had problems with the relaxers used (at least once), 75% of whom decided to stop. Half of those who stopped resumed the use of relaxers almost immediately or after a period of time. The problems encountered included hair loss, burns to the scalp, and color change. Surprisingly fake/counterfeit products were not implicated as a cause of the problems experienced, although they are known to be on the market. CONCLUSION: Despite the problems encountered, a significant number of Kenyan women in this region have used and continue to use relaxers. PMID- 17919203 TI - Alopecia in consultations in the dermatology department at Burkina Faso: epidemiologic, clinical, and etiologic aspects. AB - BACKGROUND: Hair loss, or alopecia, in a man or woman can have major psychologic repercussions. AIM: The aim of this study was to describe the epidemiologic, clinical, and etiologic aspects of alopecia in our service. MATERIALS AND METHODS: A retrospective study was performed over two years. All patients of both sexes, who consulted a dermatologist for alopecia, were included. Sociodemographic, clinical, and etiologic data were collected. RESULTS: The prevalence of alopecia is 1.02%. It generally concerns young people who are pupils or students. Hair loss was generally asymptomatic; the onset was often progressive. Alopecia was diffuse in 13.20% of cases. It was noted that 13.20% involved partial alopecia. The scalp was scarred or inflamed in 54.71% of cases. Tinea (21 cases), Alopecia areata (14 cases), Keloid folliculitis (6 cases), androgenetic alopecia (4 cases), traction alopecia (4 cases) and cosmetic alopecia (2 cases) were the most common etiologies we found in our patients. CONCLUSION: This preliminary study shows few epidemiologic, clinic and etiological aspects of the alopecia in Black African. PMID- 17919202 TI - Skin cancers at the National University Hospital of Cotonou from 1985 to 2004. AB - BACKGROUND: Skin cancer is the most common form of cancer occurring in adults in Europe and the USA. A low incidence has been reported in the black American population and in Africa, however. This study investigates the incidence of melanoma and epidermoid and basocellular carcinoma at Cotonou in Benin. METHODS: Over 20 years, 19 patient records were collected, 16 of which were the subject of a retrospective epidemiologic study. RESULTS: The main characteristic of skin cancer in the Department of Dermatology, Cotonou, Benin was its extreme rarity. The frequency was 0.00066%, with a male to female ratio of 1.28 and a mean age of 54.81 years. The mean ages at diagnosis for epidermoid carcinoma, melanoma, and basocellular carcinoma were 61.66, 59.40, and 54.56 years, respectively. The mean duration of epidermoid carcinoma was 6-10 times longer than that of melanoma and basocellular carcinoma. Of the 16 patients included in the study, five were albinos. In this group, the frequency was 31.25%. DISCUSSION: The extreme rarity of skin cancers in our series cannot be the result of recruitment bias alone. It reflects the actual situation. All of our cases of melanoma were in a plantar location. This inconsistency with other studies is probably a result of the size of our series. The duration of disease in our cases of carcinoma was comparable with the results obtained in a previous study in Dakar, Senegal. In our study, oculocutaneous albinism was the main preneoplastic factor. CONCLUSION: This study confirms the low incidence of skin cancer in the black population, the fact that basocellular carcinoma affects a relatively young population, and the predilection of melanoma for acral locations in black individuals. PMID- 17919204 TI - Mycosis fungoides and Sezary syndrome against a human immunodeficiency virus positive background: case report. AB - BACKGROUND: Mycosis fungoides (MF) and Sezary syndrome (SS) are forms of primary cutaneous T-cell lymphoma. They are relatively rare diseases, infrequent in Europe, Asia, and Africa. In Rwanda, they remain very rare. Their etiopathogenesis has long been obscure and poorly understood. Recent studies, however, have confirmed that they are of viral origin, specifically human T-cell lymphotropic virus type I (HTLV-I) and human immunodeficiency virus (HIV). Their incidence is low: about three new cases per year per million inhabitants. Individuals affected are usually aged between 40 and 60 years and of male gender. The incidence of SS is less than one new case per year per million inhabitants. METHODS: A new case of MF/SS syndrome against a HIV-positive background is described and a literature review is performed. RESULTS: Clinically, MF is characterized at onset by pruriginous, disseminated, erythematous, scaly plaques, localized on the skin, which gradually become infiltrative. In advanced stages, the nodular lesions can become ulcerated and Sezary cells are detected in the peripheral blood. Our male Rwandan MF/SS/HIV-positive patient presented with pruriginous, erythematous, scaly lesions, which had developed over 4 years and were now infiltrative, together with nodular lesions of various sizes, localized on the face, scalp, and palmar and plantar regions. All of these clinical signs, the anatomopathologic examination, retroviral serology, and testing of peripheral blood confirmed the diagnosis of MF/SS against a background of immunodepression. CONCLUSIONS: MF and SS are relatively rare cutaneous lymphomas, but have been diagnosed in Rwanda against a clinical picture of immunodepression. Their etiology seems to be viral in origin. MF and SS are difficult diseases to diagnose, with confirmation only at advanced stages. No tests exist to aid and confirm the diagnosis in the initial stages of the diseases. Further studies are urgently needed on these diseases. PMID- 17919205 TI - Epidemiologic, clinical, and therapeutic features of acne in Dakar, Senegal. AB - BACKGROUND: Acne, a chronic inflammatory dermatosis of the pilosebaceous follicle, is very widespread worldwide, affecting 75-80% of adolescents, with an impact on their quality of life. The etiologic factors are not always the same, and the treatments proposed are not always well tolerated in black individuals. AIMS: To determine how acne affects the black population in Dakar, Senegal, and to assess the results of and tolerance to treatment. METHODS: This was a prospective study conducted on a sample of patients who attended consultations for acne in our dermatology department between December 2002 and March 2003, and who agreed to take part in the study (informed consent was obtained). An anonymous questionnaire was used as a support. RESULTS: Ninety-three patients were included in the study, with a hospital frequency of 5.3% and an average age of onset of acne of 25.58 years. In 75% of cases, the acne sufferers were young women, 76.3% of whom were single. Acne was of the superficial inflammatory type in 76.3%, retentional in 67.7%, and nodulocystic in 12.9% (Some patients who have acne vulgaris may have pustules, papules and nodules or cystic lesions so the percentages add up to 100). Signs of hyperandrogeny were observed in 19.1%, with hirsutism in 93.8%. Artificial depigmentation was noted in 38.7%. Stress was found in 89.2% and a depressive state in 3.4%. Solar exposure aggravated the lesions in 52.7%. In terms of therapy, only 1.1% received oral retinoids. An improvement was noted in 67.2%, and resistance was observed in 7.3%. DISCUSSION: In Dakar, Senegal, patients with acne consult later than elsewhere. The majority of cases are women, contrary to certain other studies. The inflammatory forms predominate in adult women. The most frequent location is on the face. An evolution to pigmentary disorders is found in 67.7%. Predisposing factors include the use of depigmenting cosmetic products and sun exposure. Local treatment, such as benzoyl peroxide and topical retinoids, which promote melanoderma in black skin, is widely used. The exorbitant cost of oral retinoids considerably limits their use in Senegal. CONCLUSION: Acne, an adolescent pathology, is increasingly being observed in adults using depigmenting cosmetics. The multiple clinical forms are dominated by inflammatory acne. The many relapses and cost of treatment have a detrimental effect on the quality of life of patients. PMID- 17919206 TI - Acne on pigmented skin. AB - Acne on pigmented skin occurs in Africans and their descendants, as well as in patients with a skin phototype above IV. Acne is common in Africans and is often the primary reason for consulting a dermatologist. Acne on pigmented skin is principally inflammatory acne. The sequelae in terms of pigmentation are often the main reason for consulting a dermatologist, and take precedence over the acne itself. Research is needed into cosmetic acne and steroid-induced dermatitis, which are common in users of skin bleaching products. The treatment for acne in pigmented skin involves the same medicines as used in acne in white skin. Of the available topical treatments, benzoyl peroxide is particularly effective for the inflammatory component. Retinoids act on both the retentional and inflammatory components of acne and have skin bleaching properties. Tetracyclines are essential on initiation of treatment, regardless of severity. They prevent an inflammatory episode at the onset of acne and rapidly improve the inflammatory component. Minocycline should not be prescribed in African patients. Fatal cases of hypersensitivity have been described, with a particular frequency in this population. Doxycycline is effective, but has photosensitizing properties. Lymecycline is well tolerated. Isotretinoin is indicated for nodular acne. The same precautions for use should be followed as in all cases of acne. The skin bleaching effects are considerable. Photoprotection should generally be imposed, but, in patients with black skin, photoprotective agents are often not applied. The administration of a cosmetic containing a skin bleaching agent combined with a photoprotective agent for application in the morning, instead of hydrating cream, is acceptable to patients, improves compliance, and is effective. PMID- 17919207 TI - Kaposi's sarcoma in a hospital setting in Lome (Togo): a study of 93 cases. AB - AIM: To define the epidemiologic and clinical profile and course of the disease in African Kaposi's sarcoma (KS) and acquired immunodeficiency syndrome (AIDS) associated KS in Togo. METHODS: This was a retrospective study performed on the medical records of patients seen at the Department of Dermatology, University Hospital of Lome, Togo from January 1994 to December 2004. The medical records of all patients with KS, who had undergone human immunodeficiency virus (HIV) serology, were included in the study. RESULTS: Ninety-three files on 98 patients with KS, who had undergone HIV serology, were included in the study. The annual incidence during the study period was 8.5. HIV serology was positive in 73 patients (78.5%) and negative in 20 patients (21.5%). The mean age of the patients with AIDS-associated KS was 33.8 +/- 8.2 years, and 49.5 +/- 15.8 years for African KS. The male to female ratio for AIDS-associated KS was 1.4, and 9 for African KS. The mortality rate at 2 years for African KS was 5%, and 45% for AIDS-associated KS. CONCLUSION: The low level of access to antiretroviral drugs in HIV-infected patients explains the morbidity and mortality from AIDS associated KS in Togo. PMID- 17919208 TI - Dermatosis papulosa nigra in Dakar, Senegal. AB - BACKGROUND: Dermatosis papulosa nigra (DPN) is a benign epithelial tumour, common in the black population. It is a special topographic form of ethnic seborrheic keratosis, which is more common in the white population. Its benign character has meant that very few studies have been performed. In Senegal, no investigation has been carried out to date. AIM: To determine the epidemiologic and semiological characteristics of DPN, and the aesthetic results of DPN exeresis. METHODS: A prospective study was carried out over a period of 6 months (November 2005 to April 2006) at the Dermatology Department, Le Dantec CHU, Dakar, Senegal. The diagnosis of DPN was clinical. If desired, exeresis of the lesions by fine-needle or fine-scissor electrosurgery was carried out, and the results were evaluated over a period of 6 weeks. RESULTS: Thirty patients were evaluated: 21 women and nine men. The average age was 39.8 years (range, 25-70 years). A family predisposition was found in 93.3% of cases. The average age of onset of the lesions was 22 years, with a progressive increase in size and number. The lesions appeared initially on the face and then spread to all photoexposed areas. The lesions were profuse, numbering between 50 and 100 in 66.6% of cases, and coalesced to form plaques in 26.6%. In other cases, the lesions were papular, pediculate, or wide-based brown to deep black in colour, and with a keratotic appearance. Eight of the 21 women practiced artificial depigmentation and presented with profuse lesions. Aesthetic problems were found in 60% of patients. When performed, exeresis was satisfactory without scarring in 60% of cases at day 45. Hypochromic scars persisted in four patients who practiced artificial depigmentation and in one who had been treated with carbon dioxide snow. CONCLUSION: This study on 30 cases of DPN is the first in Senegal. The classic female predominance, family predisposition, and photodistribution of the lesions were found. The lesions were profuse, with a background of artificial depigmentation. This suggests that the sun may be an etiopathogenic factor. Aesthetic problems are an indication for ablation of the lesions using a method that minimises the risk of scarring. Artificial depigmentation may cause delayed healing and dyschromic scars, as may the application of carbon dioxide snow. PMID- 17919209 TI - Some Nigerian plants of dermatologic importance. AB - BACKGROUND: According to the World Health Organization (WHO), 80% of the world's population uses medicinal plants in the treatment of diseases and, in African countries, this rate is much higher. In recent years, however, medicinal plants have represented a primary health source for the pharmaceutical industry. No less than 400 compounds derived from plants are currently used in the preparation of drugs, such as vincristine and vinblastine used in the treatment of cancer. Nigerians still depend largely on crude herbal remedies or traditional medicine. They also use wild plants for cosmetics and perfumery. Some of these herbal remedies have been observed to be effective in certain skin diseases. METHODS: The data were obtained from history questionnaires completed by patients at the Dermatology Clinic, Lagos State University Teaching Hospital (LASUTH), Ikeja, Lagos, Nigeria, and from oral interviews with vendors and prescribers of herbal preparations at major markets at Lagos and Ijebu-Ode in south-west Nigeria, between July 2004 and July 2006. Photographs of plants were taken at private residences at Lagos, Ibadan, and Ijebu-Ode in south-west Nigeria. A literature search was conducted on 38 of the plants. The data are presented in tabular form. RESULTS: Sixty-five per cent of patients had applied some form of herbal remedy before attending our clinic. The reasons for consultation included relapses, unsustained relief, incomplete resolution, and post inflammatory hyperpigmentation. Lesions for which herbs were successfully applied included infantile eczema and seborrhoiec dermatitis, atopic eczema, impetigo, impetiginized eczema, tinea capitis, scabies, erythema multiforme, leg ulcers, localized vitiligo, and sexually transmitted diseases. Partial relief was achieved in dermatophytoses, ichthyosis, leprosy, and systemic lupus erythematosus (SLE). Some forms of alopecia, onychomycosis, and vitiligo, as well as allergic dermatoses, were not improved by herbal medicines. The preparation of the remedies was mostly by heating and boiling, infusion, and maceration. In most cases, mixtures of plants or other substances were used. The preparations were applied as poultices, ointments, baths, soaks and soaps, and oral fluids. The plants used included Adansonia digitata, Aframomum melegueta, Aloe species, Azadirachta indica, Cassia alata, Alstonia boonei, Ficus asperifolia, Cocos nucifera, Jatropha gossypyfolia, Ocimum gratissimum, Ricinus communis. A literature search on 38 of the plants used by herbal prescribers revealed the presence of established antimicrobial agents, immune modulating agents, antioxidants, other vitamins and minerals, volatile oils and emollients, and anti inflammatory agents. Some of the plants may be contaminated by mycotoxins because of poor storage. CONCLUSION: There appears to be clinical, scientific, and pharmacologic basis for the use of herbal preparations. Nigeria needs to provide effective coordination of the practice of herbal medicine to ensure safety, standardization, and preservation of the flora. PMID- 17919210 TI - Propofol in emergency medicine: further evidence of safety. PMID- 17919211 TI - Review of dyspnoea quantification in the emergency department: is a rating scale for breathlessness suitable for use as an admission prediction tool? AB - Acute shortness of breath is a potential marker of serious cardiopulmonary disease and requires rapid assessment. In our current health-care system, increasing pressure on the ED to limit costs and waiting times has resulted in the development of many clinical decision aids and admission prediction tools designed to assist ED physicians in meeting these demands. However, most of these tools are disease specific, and none are currently available for application to patients presenting to the ED with shortness of breath. Although somewhat limited, current evidence supports the utilization of a simple dyspnoea rating scale, to assist in the streamlining of clinical severity assessments and urgency evaluations, and to potentially provide useful information to facilitate rapid and accurate site-of-care decisions in this setting. PMID- 17919212 TI - Profiling adverse respiratory events and vomiting when using propofol for emergency department procedural sedation. AB - OBJECTIVES: To evaluate the rate of adverse respiratory events and vomiting among ED patients undergoing procedural sedation with propofol. METHODS: This was a prospective, observational series of patients undergoing procedural sedation. Titrated i.v. propofol was administered via protocol. Fasting status was recorded. RESULTS: Four hundred patients undergoing sedation were enrolled. Of these 282 (70%, 95% confidence interval [CI] 66-75%) had eaten or drunken within 6 and 2 h, respectively. Median fasting times from a full meal, snack or drink were 7 h (interquartile range [IQR] 5-9 h), 6 h (IQR 4-8 h) and 4 h (IQR 2-6 h), respectively. Overall a respiratory event occurred in 86 patients (22%, 95% CI 18 26%). An airway intervention occurred in 123 patients (31%, 95% CI 26-35%). In 111 cases (90%, 95% CI 60-98%) basic airway manoeuvres were all that was required. No patients were intubated. Two patients vomited (0.5%, 95% CI 0.0 1.6%), one during sedation, one after patient became conversational. One patient developed transient laryngospasm (0.25%, 95% CI 0-1.2%) unrelated to vomiting. There were nil aspiration events (0%, 95% CI 0-0.74%). CONCLUSIONS: Seventy per cent of patients undergoing ED procedural sedation are not fasted. No patient had a clinically evident adverse outcome. Transient respiratory events occur but can be managed with basic airway interventions making propofol a safe alternative for emergency physicians to provide emergent procedural sedation. PMID- 17919213 TI - Optimization of propofol dose shortens procedural sedation time, prevents resedation and removes the requirement for post-procedure physiologic monitoring. AB - OBJECTIVE: To examine the effect of propofol dosing (total dose and number of doses) on patient sedation time and likelihood of resedation. METHODS: This was a prospective, observational patient series in an urban district hospital ED with 42 000 attendances per annum. Patients undergoing an emergent procedure requiring procedural sedation were included. Titrated intravenous propofol was administered according to departmental procedure. Standardized consent and data collection forms were used. Time taken for the patient to become conversational after first administration was recorded and any resedation documented. RESULTS: Four hundred patients, undergoing 404 procedures, were enrolled for the period commencing August 2004 until March 2006. The mean initial propofol bolus was 0.8 mg/kg (SD 0.6), and mean total propofol dose was 1.8 mg/kg (SD 1.0), comprising a mean of 2.3 (SD 2.1) doses of 15.8 mg (SD 11.4). Mean sedation time was 11.8 min (SD 6.9), and increased sedation times were associated with higher total propofol dose and number of boluses (P < 0.0001). Resedation occurred in two patients (0.5%, 95% CI 0-1.6%). CONCLUSION: Shorter sedation times are seen with lower doses of propofol. Patients do not need prolonged post-procedure monitoring because the occurrence of spontaneous resedation associated with propofol use is a rare event. This has implications for patient flow and staff resource allocation in a busy ED. PMID- 17919214 TI - Identifying severe community-acquired pneumonia in the emergency department: a simple clinical prediction tool. AB - OBJECTIVE: To identify independent predictors of severe pneumonia in a local population, and create a simple severity score that would be useful in the ED. METHODS: Data on the clinical features of patients presenting to hospital with community-acquired pneumonia were collected. Multivariate logistic regression was used to identify independent predictors of death, requirement for ventilatory or inotropic support, and these combined. These predictors were used to modify an existing severity score, and its performance was tested in a second cohort of patients. RESULTS: A total of 392 patients in the derivation, and 330 in the validation cohorts. Independent predictors of 'death and/or requirement for ventilatory or inotropic support' were: systolic blood pressure (BP) <90 mmHg (OR 3.49 [95% CI 1.12-10.38]); acute confusion (OR 5.48 [95% CI 2.74-10.99]); oxygen saturations < or =90% (OR 3.49 [95% CI 1.77-6.89]); and respiratory rate > or =30/min (OR 2.65 [95% CI 1.35-5.21]). Age >65 years was not an independent predictor in this patient group (OR 0.52 [95% CI 0.23-1.16]). This information was used to propose that severe pneumonia could be predicted by two or more of: acute confusion; oxygen saturations < or =90%; respiratory rate > or =30/min; and either systolic BP <90 mmHg; or diastolic BP < or =60 mmHg. In a separate cohort, the performance of this score was similar to other tools. CONCLUSION: This provides a practical tool that can be used to 'flag' impending patient demise. Its advantages are that it is simple, uses predictive variables, does not require invasive testing, and removes bias regarding patient age. Like other tools, its accuracy is not perfect, and it should only be used to augment clinical judgement. PMID- 17919215 TI - Lignocaine is a better analgesic than either ethyl chloride or nitrous oxide for peripheral intravenous cannulation. AB - BACKGROUND: Peripheral intravenous (i.v.) cannulation is a painful, frequently performed ED procedure. It is common practice in other medical settings to offer analgesia prior to cannulation. OBJECTIVE: The present trial aims to reproduce in the ED studies that found a reduction in the pain of i.v. cannulation after intradermal lignocaine, ethyl chloride topical spray and entonox (50:50 oxygen : nitrous oxide). It also intends to determine which is analgesic most effective and explore the role of entonox for cannulation analgesia. METHODS: Three hundred subjects were randomized into four groups: i.v. cannula inserted with (i) no anaesthesia; (ii) entonox; (iii) ethyl chloride; and (iv) 0.1 mL intradermal 1% lignocaine. Pain was recorded on 100 mm visual analogue scales (VAS) after lignocaine injection or ethyl chloride spray and following i.v. cannulation. A clinically significant reduction in VAS pain score was determined to be 13 mm. RESULTS: Patients cannulated without analgesia reported the most pain. Those cannulated after lignocaine had the least pain (median VAS 20 mm, 95% CI 15-25, vs 1 mm 95% CI 0-6, P < or = 0.001). Ethyl chloride (VAS 11 mm, 95% CI 7-15, P = 0.003) and entonox (VAS 13 mm, 95% CI 8-18, P = 0.047) reduced i.v. cannulation pain but did not reach clinical significance. Neither pain from presenting symptoms (P = 0.3), nor size of cannula (P = 0.8) affected pain scores. VAS scores were independent of sex and age (P = 0.1). Cannulation success was not affected by either the choice of analgesia or cannulation site. CONCLUSIONS: The present trial confirms the findings of Harris and colleagues that lignocaine reduces the pain of cannulation in the ED. Lignocaine reduced the pain of i.v. cannulation more effectively than entonox or ethyl chloride. PMID- 17919216 TI - Handover in the emergency department: deficiencies and adverse effects. AB - OBJECTIVE: To determine problems resulting from ED handover, deficiencies in current procedures and whether patient care or ED processes are adversely affected. METHODS: A prospective observational study at three large metropolitan ED comprising three components: observation of handover sessions, 2 h post handover surveys of the receiving doctors and a general survey of ED doctors. RESULTS: The handovers of 914 patients were observed during 60 handover sessions in a 3-month period. Medical information, including presenting complaints, was handed over better than communication and disposition information. Seven hundred and seven (77.4%) of 914 potential post-handover interviews were undertaken. Most (88.3%) doctors thought the handover was 'adequate/good'. However, information was perceived as lacking in 109 (15.4%) handovers, especially details of management (35, 5.0%), investigations (33, 4.7%) and disposition (33, 4.7%). There was a significant difference in the perceived quality of handovers (1-5 scale where 5 = excellent) when all required information was handed over and when it was not (median scores 4.0 vs 3.0, respectively, P < 0.001). As a result of perceived inadequate handovers, the doctor/ED and patient were affected adversely in 62 (8.8%) and 33 (4.7%) cases, respectively, for example, repetition of assessment, delays in disposition and care. Fifty doctors completed the general survey. Most believed communications made to inpatient units, inaccurate/incomplete information and disorganization were problematic. CONCLUSION: Deficiencies in handover processes exist, especially in communication and disposition information. These affect doctors, the ED and patients adversely. Recommendations for improvement include guideline development to standardize handover processes, the greater use of information technology facilities, ongoing feedback to staff, and quality assurance and education activities. PMID- 17919217 TI - Accuracy of ECG electrode placement by emergency department clinicians. AB - OBJECTIVES: Misplaced ECG electrodes can cause changes in ECG recordings, which could have an impact on clinical decisions. We aimed to determine the inter-rater reliability of ECG electrode placement by senior clinical staff in the ED. METHODS: A prospective observational study was conducted in adult patients undergoing an ECG as part of their routine ED care. Adhesive electrodes were left in place after an ECG had been performed by the treating nurse, and subsequently each patient was assessed by two of the three investigators. Each investigator independently recorded the location of the chest electrodes relative to the recommended standard positions. Displacement of the electrodes from the standard positions was measured in the vertical and horizontal planes. The age, sex, weight, height and chest circumference was also recorded. Comparisons were made between investigators to determine variability in assessment of the standard positions. RESULTS: Measurement of horizontal and vertical displacement for each of the six chest leads in the 77 patients resulted in 924 paired measurements. There was substantial inter-rater variation in the measurement of both vertical (mean 13.5 mm, range 0-105 mm) and horizontal (mean 16.5 mm, range 0-120 mm) displacement. This variation was greater in the lateral chest leads and was more marked in women than in men, especially in the vertical plane (lead V6: men 14.5 mm vs women 27.0 mm, P < 0.01). CONCLUSION: Among clinical 'experts', there is wide variation in the identification of the correct location for electrode placement, particularly in the lateral leads and in women. This has significant implications when comparing ECG in which electrodes have been placed by different clinicians. PMID- 17919219 TI - Route of administration of redback spider bite antivenom: determining clinician beliefs to facilitate Bayesian analysis of a clinical trial. AB - OBJECTIVE: To determine current beliefs of Australasian emergency physicians, to form the basis of 'stopping rules' for a clinical trial of intravenous (i.v.) versus intramuscular (i.m.) redback spider antivenom. METHODS: An email survey of fellows and trainees of the Australasian College for Emergency Medicine. RESULTS: There were 218 responses; 30% used the i.v. route exclusively, 16% used the i.m route exclusively, 17% used i.m. followed by i.v. if there was a poor initial clinical response, and 38% stated that they had no particular preference. The probability given by respondents that the i.v. route is superior allowed us to differentiate 'i.v. enthusiasts' from 'i.v. sceptics'. Median predicted response rates were 90% versus 80% for the i.v. route and 60% versus 75% for the i.m. route in the enthusiastic and sceptical groups, respectively. The median expected absolute advantage of i.v. compared with i.m. antivenom was 20% versus 5%, respectively. The median number-needed-to-treat threshold that would lead respondents to choose the i.v. route in preference to the i.m. was 5. CONCLUSION: Australasian emergency physicians have polarized views on the optimal route for administering redback spider antivenom. We were therefore able to define both sceptical and enthusiastic priors for a fully Bayesian trial analysis. Our findings support using a number needed to treat of 5 (20% absolute advantage) for powering a clinical study and for determining the point at which it should be stopped. PMID- 17919218 TI - Management and outcome of spontaneous pneumothoraces at three urban EDs. AB - OBJECTIVE: Spontaneous pneumothoraces predominantly affect young people. Substantial morbidity arises from the disease, invasive procedures and hospitalization. The literature is inconsistent regarding optimal management. This retrospective study aimed to define factors affecting the outcome of these patients. METHODS: Patients were identified from databases at three EDs for explicit retrospective medical record review. Iatrogenic and traumatic pneumothoraces were excluded. Data collected included demographic details, treatment and outcome. The primary outcome was failure of initial treatment, defined as the need for a second treatment modality (including inpatient pleurodesis for persistent air leak) or re-presentation within 5 days of treatment cessation. Associations with the primary outcome in primary spontaneous pneumothorax were assessed using logistic regression. RESULTS: One hundred and twenty-one spontaneous pneumothoraces were identified. There was poor correlation between clinician estimates and objective measurement of pneumothorax size. Pneumothorax size, measured using the average interpleural distance method, was the only independent predictor of treatment failure. Initial treatment modality demonstrated a confounder relationship with outcome. Subgroup analysis for patients treated with continuous pleural drainage compared small and large calibre drainage tubes, with no significant difference found. CONCLUSIONS: Objective measurement of pneumothorax size was the only independent predictor of treatment failure, with initial treatment modality having a confounding effect. Algorithms regarding initial treatment modalities are usually based on pneumothorax size and presence or absence of symptoms. The present study illustrates the importance of objective assessment of pneumothorax size in both clinical research and clinical practice. PMID- 17919220 TI - Investigation of the rate of meningitis in association with urinary tract infection in infants 90 days of age or younger. AB - OBJECTIVE: To test the hypothesis that urinary tract infections (UTI) in young infants are rarely associated with meningitis. METHODS: We undertook a review of the laboratory results from 322 infants, 90 days of age or younger, with an admission or discharge diagnosis of UTI or meningitis. The study was conducted in a tertiary paediatric hospital. The primary outcome measure was the incidence of coexisting urinary tract and cerebrospinal fluid sepsis. RESULTS: In total, 161 of the 322 (50%) infants with an admission or discharge diagnosis of UTI or meningitis were subsequently shown to have a culture-proven UTI. Of the children with a culture-proven UTI, 75 (47%) had cerebrospinal fluid obtained. We detected one case of probable bacterial meningitis in association with UTI. CONCLUSION: UTI is rarely associated with meningitis in infants 90 days of age or younger. PMID- 17919221 TI - Emergency medicine in a developing country: experience from Kilimanjaro Christian Medical Centre, Tanzania, East Africa. AB - Tanzania in East Africa has a population of over 36 million and is one of the poorest countries in the world. Life expectancy has declined and infant mortality rates are increasing. Four consultant specialist hospitals and 17 regional hospitals service the mainland. Kilimanjaro Christian Medical Centre is a major specialist teaching hospital with 500 beds, serving the entire north-west of the country. There is a small 'casualty' ward with three cubicles and one resuscitation room. Malaria, HIV, respiratory infections and gastroenteritis are the chief causes of death in children. Changing lifestyle and Western influences have increased diabetes and vascular disease in adults, and large numbers of trauma deaths are increasingly encountered. Kilimanjaro Christian Medical Centre 'Casualty' admission data are presented, as well as an insight into the challenges of emergency medicine in this country. PMID- 17919222 TI - Stephen's Banded Snake envenomation treated with tiger snake antivenom. AB - Demonstration of the use of Tiger Snake Anti-venom in Stephen's Banded Snake envenomation is described. The patient presented with a clear history of a bite and a mild headache. Subsequently, the patient developed defibrination coagulopathy. A swab of the bite site reacted to the Tiger Snake Antivenom Indicated Well 1 [corrected] on the Commonwealth Serum Laboratories Snake Venom Detection Kit [corrected] Two ampoules of anti-venom were used. A mild allergic reaction to tiger snake antivenom developed. There was resolution of the coagulopathy. PMID- 17919223 TI - Air bag-associated burn. AB - Burns due to the deployment of air bags have been occasionally described in recent years. Most reports are about injuries in the USA. However, there have been few reports of this type of injury in Australia. This case report details such an injury that took place in a road traffic accident in Melbourne, Australia. PMID- 17919224 TI - Magnesium sulphate for phaeochromocytoma crisis. PMID- 17919226 TI - Treating nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease (NAFLD) is a chronic illness with multiple consequences. The spectrum of disease ranges from simple steatosis, with benign prognosis, to a potentially progressive form, nonalcoholic steatohepatitis, which may lead to liver fibrosis and cirrhosis, leading to an increase in morbidity and mortality. Furthermore, hepatocellular carcinoma incidence in NAFLD is comparable with that observed in hepatitis C-infected patients once cirrhosis is established. Current therapy is limited to lifestyle changes and control of associated metabolic disorders; however, new treatments are on the way from basic research to bedside. A review of the current literature on treatment of nonalcoholic fatty liver disease is presented in this article. PMID- 17919227 TI - Percutaneous liver biopsy in clinical practice. AB - Percutaneous liver biopsy (PLB) is the standard procedure for obtaining hepatic tissue for histopathological examination, and remains an essential tool in the diagnosis and management of parenchymal liver diseases. The use of liver biopsy (LB) is increasing with the advent of liver transplantation and the progress being made in antiviral therapeutic agents. While blind percutaneous needle biopsy is the traditional technique, the use of ultrasound (US) guidance has increased considerably. Literatures were reviewed to assess the existing clinical practice of PLB with an emphasis on the technique, the operator, types of biopsy needles, quality of LB specimens and the risk of complications. The best available evidence indicates that the use of ultrasound-guided biopsy (UGB) is superior to blind needle biopsy (BNB). The odds ratios of the controlled studies showed that BNB carried a higher risk for major complications, postbiopsy pain and biopsy failure. Therefore, percutaneous LB under US control is superior to BNB and it is recommended that UGB be considered the standard of care for this important and widely used invasive procedure in the field of clinical hepatology. PMID- 17919228 TI - Immunohistochemical analysis of the progression of flat and papillary preneoplastic lesions in intrahepatic cholangiocarcinogenesis in hepatolithiasis. AB - BACKGROUND: Two types of precursor lesions, flat-type 'biliary intraepithelial neoplasia (BilIN)' and papillary-type 'intraductal papillary neoplasm of the bile duct (IPNB)', are proposed in the tumorigenesis of intrahepatic cholangiocarcinoma (ICC) in hepatolithiasis. METHODS: In this study, the participation of cancer-related molecules in the progression of these two precursor lesions was examined, using 64 hepatolithiatic livers with BilIN lesions (45 livers) and IPNB lesions (19 livers) and 10 hepatolithiatic livers without neoplastic lesions as a control. The expression of E-cadherin, beta catenin, matrix metalloproteinase-7 (MMP-7), membrane type 1-MMP (MT1-MMP), cyclin D1 and c-myc was immunohistochemically examined. RESULTS: The membranous expression of beta-catenin decreased along with the progression in both BilIN and IPNB lineages. Membranous expression of E-cadherin was significantly decreased in invasive ICC with BilIN and IPNB in comparison with non-invasive counterparts. MMP-7 and MT1-MMP were commonly expressed in invasive ICC with BilIN (100%), while non-invasive lesions (BilIN-1, -2, -3) and the IPNB lineage were only occasionally and weakly positive for these molecules. Cyclin D1 and c-myc, target molecules of Wnt signalling, were frequently positive in the IPNB lineage (65 and 54% respectively), and interestingly nuclear beta-catenin staining, reflecting activation of Wnt signalling, was observed only in the IPNB lineage (22%) (P<0.05). CONCLUSIONS: Decreased membranous expression of beta-catenin and E cadherin is an early event in the tumorigenesis of both BilIN and IPNB lineages. The expression of MMP-7 and MT1-MMP was closely associated with invasive growth of the BilIN lineage. The Wnt signalling pathway may play an important role in the tumorigenesis of the IPNB lineage. PMID- 17919230 TI - Malnutrition and diabetes mellitus are related to hepatic encephalopathy in patients with liver cirrhosis. AB - BACKGROUND/AIMS: Studies on animal models of hepatic encephalopathy (HE) suggest that poor nutritional status may facilitate the development of HE. Insulin resistance and diabetes mellitus have recently been reported to affect cognition in patients with hepatitis C cirrhosis awaiting liver transplantation. Our aim was to investigate the effects of malnutrition and diabetes mellitus on HE in unselected patients with liver cirrhosis. METHODS: A total of 128 consecutive cirrhotic patients were prospectively evaluated for the presence of HE according to the West-Haven criteria as well as by means of two psychometric tests and fasting plasma ammonium ion concentrations. Nutritional status was assessed by anthropometry and estimation of recent weight change. Fasting plasma glucose was measured, and in a subgroup of 84 patients fasting serum insulin and insulin resistance were also determined. RESULTS: Fifty-one (40%) cirrhotics were malnourished, 33 (26%) had diabetes and 42 (34%) had HE. Patients with vs. without malnutrition had more frequently HE (46 vs. 27%; P=0.031) but did not differ in age, aetiology or severity of liver cirrhosis (P>0.1). Multivariate analysis showed that the time needed to perform number connection test A was independently correlated to age, the Child-Pugh score, diabetes and malnutrition (P<0.05 for all). Plasma ammonium ion levels were related to insulin resistance (r=0.42, P<0.001) and muscle mass (r=0.28, P=0.003). CONCLUSION: Malnutrition and diabetes mellitus seem to be related to HE in patients with liver cirrhosis. Nutritional status and insulin resistance might be implicated in the pathogenesis of HE. PMID- 17919229 TI - Interferon and lamivudine vs. interferon for hepatitis B e antigen-positive hepatitis B treatment: meta-analysis of randomized controlled trials. AB - AIMS: To compare interferon monotherapy with its combination with lamivudine for hepatitis B e antigen (HBeAg)-positive hepatitis B treatment. METHODS: Two independent researchers identified pertinent randomized controlled trials. The trials were evaluated for methodological quality and heterogeneity. Rates of sustained virological and biochemical responses, and HBeAg clearance and seroconversion were used as primary efficacy measures. Quantitative meta-analyses were conducted to assess differences between groups for conventional and pegylated interferon, and overall. RESULTS: Greater sustained virological, biochemical and seroconversion rates were observed with addition of lamivudine to conventional [odds ratio (OR)=3.1, 95% confidence intervals (CI) (1.7-5.5), P<0.0001, OR=1.8, 95% CI (1.2-2.7), P=0.007 and OR=1.8, 95% CI (1.1-2.8), P=0.01 respectively], although not pegylated [OR=1.1, 95% CI (0.5-2.3), P=0.8, OR=1.0, 95% CI (0.7-1.3), P=0.94, and OR=0.9, 95% CI (0.6-1.2), P=0.34 respectively] interferon-alpha, with no significant affect on HBeAg clearance rates [OR=1.6, 95% CI (0.9-2.7), P=0.09, and OR=0.8, 95% CI (0.6-1.1), P=0.26 respectively]. Excluding virological response (P<0.001), pegylated interferon monotherapy and conventional interferon and lamivudine combination therapy were similarly efficacious (P>0.05), with the former studied in harder to treat patients, as evidenced by the superior virological response observed with conventional as compared with pegylated interferon monotherapy (P<0.0001). CONCLUSION: In comparable populations, pegylated interferon monotherapy is likely to be equally or more efficacious than conventional interferon and lamivudine combination therapy, thus constituting the treatment of choice, with no added benefit with lamivudine addition. However, when conventional interferon is used, its combination with lamivudine should be considered. PMID- 17919231 TI - Recipient cytotoxic T lymphocyte antigen-4 +49 G/G genotype is associated with reduced incidence of hepatitis B virus recurrence after liver transplantation among Chinese patients. AB - BACKGROUND: Single-nucleotide polymorphisms (SNP) of two-gene locus cytotoxic T lymphocyte antigen-4 (CTLA-4) +49 and CD86 +1057 were previously reported to influence the outcome of liver transplantation (LT) with respect to allograft acceptance. SNP at CTLA-4 +49 was also suggested to be associated with the individual difference in the clearance of hepatitis B virus (HBV). However, their influence on the incidence of post-LT HBV reinfection was not clear. With the increasing knowledge of costimulatory mechanisms on LT and host immune response, we designed this study to investigate the relationship between different alleles as well as genotypes at these two locations and HBV reinfection after LT. METHODS: Genomic DNA from 167 LT recipients with HBV-related diseases was genotyped for CTLA-4 +49 and CD86 +1057 genomic polymorphisms using a sequence specific primer-polymerase chain reaction (PCR-SSP). HBV recurrence was diagnosed based on the serological and pathological finding of HBV DNA and HBsAg. RESULTS: The present study indicated that the recipients with CTLA-4 +49 GG genotype had a reduced risk (6.67%) of HBV recurrence compared with non-CTLA-4 +49 GG-carrying individuals (20.7%) (relative risk 3.098) (P=0.032). The allelic frequency of CTLA-4 +49 G was also significantly lower in patients with HBV recurrence, compared with that in patients without HBV recurrence (P=0.013, odds ratio 2.176, 95% confidence interval 1.170-4.046). However, no significant association was found between CD86 +1057 and HBV recurrence. CONCLUSION: Our result on CTLA-4 +49 A/G polymorphism indicated that the CTLA-4 +49 GG genotype was related to a reduced risk in the incidence of HBV recurrence. PMID- 17919232 TI - Similar effects of recombinant interferon-alpha-2b and natural interferon-alpha when combined with intra-arterial 5-fluorouracil for the treatment of advanced hepatocellular carcinoma. AB - AIM: Intra-arterial 5-fluorouracil (5-FU) plus interferon (IFN) combination therapy is effective against advanced hepatocellular carcinoma (HCC) with portal vein tumour thrombosis. In this study, we compared the efficiency and safety of recombinant IFN-alpha-2b with natural IFN-alpha as components of the combination therapy. METHODS: Consecutive HCC patients (n=31) with portal vein tumour thrombosis were enrolled in this prospective study. They received combination therapy of 5-FU and either recombinant IFN-alpha-2b (R group, n=15) or natural IFN-alpha (N group, n=16). We compared the two groups for the early response rate, adverse reactions, time to progression (TTP) and survival rates. In addition, we assessed the cost-effectiveness of each protocol. RESULTS: The early response rate (R: 26.7%, N: 31.2%), median TTP (R: 5.8 months, N: 5.6 months) and median survival time (R: 7.5 months, N: 6.5 months) were not significantly different between the R and N groups. There were no differences in adverse reactions between the two groups. The estimated cost-effectiveness ratio of recombinant IFN-alpha-2b was better than natural IFN-alpha. CONCLUSIONS: In our protocol of combination therapy, there were no significant differences between recombinant IFN-alpha-2b and natural IFN-alpha with regard to early response to therapy, adverse effects, TTP and survival rates. 5-FU could be combined with either recombinant IFN-alpha-2b or natural IFN-alpha, although the cost effectiveness of the former warrants its use clinically. PMID- 17919234 TI - Antimitochondrial antibodies of immunoglobulin G3 subclass are associated with a more severe disease course in primary biliary cirrhosis. AB - BACKGROUND/AIMS: Primary biliary cirrhosis (PBC) is characterised by the presence of immunoglobulin (Ig) G antimitochondrial antibodies (AMA), which are routinely detected by indirect immunofluorescence (IFL) using composite rodent tissue substrate. The IgG subclass distribution and clinical significance of IFL detected AMA in patients with PBC have not been previously studied in detail. METHODS: We have examined IgG subclass-specific AMA detected by IFL on rodent liver, kidney and stomach tissue substrate using affinity-purified IgG subclass monospecific antisera as revealing reagents in 95 AMA-positive PBC patients from Greece. RESULTS: AMA of any of the IgG1, IgG2 or IgG3 subclasses were present in 89/95 (93.7%) patients. Among those 89, 55 (61.8%) had IgG1, 2, 3 AMA positivity; eight (9%) had IgG1, 2; seven (7.9%) had IgG2, 3; eight (9%) had IgG1, 3; nine (10.1%) had IgG1 subclass and two (2.2%) single IgG3 AMA reactivity. IgG4 AMA was absent. IgG3 titres were higher than IgG2 and IgG1 (P<0.001) and IgG1 higher than IgG2 (P<0.001). IgG3 AMA-positive patients had a histologically more advanced disease (P<0.01) and were more frequently cirrhotic compared with those who were negative (P<0.01). There was a positive correlation between AMA IgG3 titre and Mayo risk score (r=0.55, P=0.009, Spearman's correlation). CONCLUSIONS: Our findings suggest that AMA are not restricted to a specific IgG subclass. AMA of the IgG3 subclass are associated with a more severe disease course, possibly reflecting the peculiar ability of this isotype to engage mediators of damage. PMID- 17919233 TI - Gamma-glutamyltransferase and rapid virological response as predictors of successful treatment with experimental or standard peginterferon-alpha-2b in chronic hepatitis C non-responders. AB - BACKGROUND: High-dose peginterferon-alpha (PegIFN-alpha) induction and prolongation of therapy may be an option to improve sustained virological response (SVR) rates among hepatitis C virus (HCV) non-responders, although a higher and a longer dosing of PegIFN-alpha may intensify side effects. METHODS: We randomized 53 patients, who previously failed with standard IFN-alpha+/ ribavirin, to a high-dose induction and an extended regimen with PegIFN-alpha-2b [3.0 microg/kg once weekly (q.w.) 12 weeks-->2.0 microg/kg q.w. 12 weeks-->1.5 microg/kg q.w. 48 weeks] or a standard regimen (1.5 microg/kg q.w. 48 weeks). All patients received daily weight-based ribavirin (800-1200 mg/day). The short-form 36 health survey was used to evaluate health-related quality of life (HRQL). RESULTS: Intention-to-treat analysis showed no significant difference in SVR rate (44% vs. 37%, P=0.62) and relapse rate (9% vs. 31%, P=0.17) between experimental and standard treatment. Overall, 80% of the [positive predictive value (PPV)] patients with rapid virological response (RVR, HCV-RNA negativity at week 4) achieved SVR. No significant dose-related differences in HRQL were seen between both groups. At baseline, genotype 2 or 3 [odds ratio (OR): 7.4, 95% confidence interval (CI): 1.4-33.3, P=0.01] and gamma-glutamyltransferase (GGT) levels <2 x ULN (upper limit of normal) (OR: 6.76, 95% CI: 1.5-31.3, P=0.009) were significantly associated with SVR. Multivariate logistic regression at week 4 showed that only baseline GGT <2 x ULN (OR: 7.3, 95% CI: 1.4-38.5, P=0.01) and RVR (OR: 15.6, 95% CI: 3.2-76.9, P<0.001) were independently predictive for SVR. CONCLUSION: Retreatment with PegIFN-alpha-2b and ribavirin for a minimum of 48 weeks should be considered in all patients unresponsive to previous IFN-based therapies. Baseline GGT values and RVR are highly predictive for retreatment outcome. PMID- 17919235 TI - Urotensin II: a novel vasoactive mediator linked to chronic liver disease and portal hypertension. AB - BACKGROUND/AIMS: Urotensin II (UII) is recognised as the most potent human vasoconstrictor; however, its role in chronic liver disease (CLD) is unknown. AIM: We sought to determine serum UII levels in CLD and explore its relationship with clinical features and outcomes of patients with CLD and portal hypertension. METHODS: UII was analysed by radio-immunoassay on cirrhotic patients undergoing hepatic venous pressure gradient (HVPG) determination and age- and sex-matched controls. Follow-up data were prospectively recorded. RESULTS: From 1997 to 2004, 80 patients (male/female: 74/6) underwent a total of 94 HVPG assessments. UII was higher in cirrhotic patients compared with controls (2.05+/-0.06 and 1.55+/-0.09 pmol/L, P<0.001) and was correlated with HVPG (r=+0.35, P=0.001) and severity of CLD (r=+0.6, P<0.001). UII was higher in patients who developed refractory ascites (2.45+/-0.13 vs. 1.7+/-0.12 pmol/L, P<0.001) and in those who died during the follow-up period (2.27+/-0.15 pmol/L vs. 1.95+/-0.08 pmol/L, P<0.05). CONCLUSION: Serum UII is elevated in patients with CLD, and is associated with the severity of the underlying liver disease and the degree of portal hypertension. Baseline levels can predict future complications such as refractory ascites and patient survival. PMID- 17919236 TI - Outcome of patients with hepatocellular carcinoma referred to a tertiary centre with availability of multiple treatment options including cadaveric liver transplantation. AB - Hepatocellular carcinoma (HCC) is a primary cancer of the liver with an established causal link to viral hepatitis and other forms of chronic liver disease. AIMS: The aim of this study was to analyse the determinants of outcome in patients with HCC referred to a tertiary centre for management. METHOD: Two hundred and thirty-five prospective patients with HCC and minimum 12-month follow up were studied. RESULTS: The cohort was heterogeneous, with 52% Caucasian, 40% Asian and 5% of Middle-Eastern origin. Independent predictors of outcome included tumour size and number, the presence of ascites or portal vein thrombosis, alpha foetoprotein >50 U/L and an impaired performance status. Treatment was determined on an individual case basis by a multidisciplinary tumour team. Surgical resection was primary treatment in 43 patients, liver transplantation in 40 patients, local ablation (percutaneous radiofrequency ablation or alcohol injection) in 33 patients, transarterial chemoembolisation in 33 patients, chemotherapy or other systemic therapy in 30 patients and no treatment in 56 patients. After adjustment for significant covariates, both liver transplantation (P<0.001) and surgical resection (P=0.029) had a significant effect on patient survival compared with no treatment, but local ablation (P=0.410) and chemoembolisation (P=0.831) did not. Liver transplantation resulted in superior overall and, in particular, disease-free survival compared with surgical resection (disease-free survival 84 vs 15% at 5 years). CONCLUSION: In conclusion, both surgical resection and liver transplantation significantly improve the survival of patients with HCC, but improvements need to be made to the delivery of loco-regional therapy to enhance its effectiveness. PMID- 17919237 TI - Glycine reduces hepatic warm ischaemia-reperfusion injury by suppressing inflammatory reactions in rats. AB - BACKGROUND: Glycine, a non-essential amino acid, is known to have an anti inflammatory effect on haemorrhagic and endotoxic shock in animals. In the present study, we examined the effects of glycine on inflammatory reactions and hepatocellular damage after hepatic warm ischaemia-reperfusion (I-R) in rats. METHODS: Using Sprague-Dawley rats, ischaemia was induced in 92% of the liver by clamping the hepatic inflows for 60 min, and part of the non-ischaemic lobe was resected after reperfusion. Before the induction of I-R, rats were treated by an intravenous administration of either glycine (Glycine group) or normal saline (Control group). The severity of hepatocellular injury was determined by serum levels of hepatic enzymes and histological necrosis. To evaluate the effect of glycine on inflammatory reactions, tumour necrosis factor (TNF)-alpha mRNA expression in the liver, serum levels of TNF-alpha and chemokine-induced neutrophil chemoattractant (CINC) and the number of neutrophils in the liver were compared between the groups. RESULTS: At 60 min after reperfusion, the serum levels of hepatic enzymes in the Glycine group were significantly lower than those in the Control group (P<0.05). TNF-alpha mRNA expression was also suppressed in the livers in the Glycine group. Furthermore, the serum levels of TNF-alpha and CINC in the Glycine group were significantly lower than those in the Control group (P<0.05). Pretreatment with glycine also significantly reduced hepatic necrosis and the number of neutrophils at 24 h after reperfusion. CONCLUSION: Glycine has a protective effect against inflammatory reactions, and reduces hepatocellular injury induced by hepatic warm I-R in rats. PMID- 17919238 TI - Hepatic stellate cell line modulates lipogenic transcription factors. AB - BACKGROUND/AIMS: Pre-adipocyte differentiation into adipocyte is a terminal differentiation process triggered by a cascade of transcription factors. Conversely, hepatic stellate cells (HSC) can switch between lipid storing and the myofibroblast phenotype in association with liver fibrotic processes. Here, adipogenic/lipogenic-related transcription factors and downstream-regulated genes were evaluated in a murine HSC cell line. GRX-HSC cells are transitional myofibroblasts that differentiate into lipocytes following retinol or indomethacin treatment. METHODS/RESULTS: Specific mRNAs were quantified by a real time polymerase chain reaction after 24 h or 7 days of cell culture with indomethacin or retinol. Proliferator-activated receptorgamma and Pex16 transcripts were increased either by retinol or indomethacin. Retinol induced a minor increase in C/enhancer binding proteinalpha transcripts, while only indomethacin increased adipsin transcripts. CONCLUSIONS: Our results showed that the myofibroblast to lipocyte phenotype switch follows partially different transcriptional pathways, according to the effector. Retinol induces lipid synthesis and storage without affecting characteristic adipocytic genes, while indomethacin treatment restores the lipocytic phenotype with increased adipisin expression. PMID- 17919239 TI - Leptin stimulates alpha1(I) collagen expression in human hepatic stellate cells via the phosphatidylinositol 3-kinase/Akt signalling pathway. AB - BACKGROUND/AIMS: Leptin has been recognized as a profibrogenic hormone in the liver and is involved in collagen type I formation by activated hepatic stellate cells (HSCs) in response to fibrogenic substances, but the molecular signal mechanisms by which leptin promotes liver fibrogenesis through upregulation of collagen type I expression is not clear. We investigated whether leptin-induced collagen type I is mediated by the Janus kinase-phosphatidylinositol 3-kinase-Akt (JAKs-PI3K-Akt) pathway in a human HSC cell line, LX-2. METHODS: LX-2 cells were treated with or without various inhibitors in the presence of leptin. RESULTS: Leptin increased alpha1(I) collagen mRNA and protein. JAK1, PI3K and Akt were activated after leptin stimulation. AG490, a JAK inhibitor, blocked JAK1 phosphorylation accompanied by inhibition of PI3K and Akt activation as well as alpha1(I) collagen mRNA expression, indicating a JAK1-dependent mechanism. Wortmannin, a PI3K inhibitor, prevented PI3K and Akt activation and resulted in suppression of alpha1(I) collagen mRNA expression, suggesting a PI3K-mediated process. These changes were reproduced by overexpression of the dominant-negative p85alpha mutant. A443654.3, an Akt inhibitor, opposed Akt activation, leading to downregulation of alpha1(I) collagen mRNA. Overexpression of the dominant negative Akt mutant led to similar alterations. CONCLUSION: Leptin has a direct action on liver fibrogenesis by stimulating alpha1(I) collagen production in activated HSC. The process appears to be mediated by the PI3K/Akt pathway through activated JAK1. PMID- 17919241 TI - Successful living donor liver transplantation using a graft from a hepatitis B surface antigen-positive donor. AB - BACKGROUND/AIMS: Liver transplantation using a graft from a donor with a positive hepatitis B surface antigen (HBsAg) has been contraindicated owing to the extremely high risk for recurrent disease leading to graft loss. However, the severe shortage of donors often forces the transplant community to utilize suboptimal donors, especially in the setting of living donor liver transplantation (LDLT). METHOD: Here, we report a case of successful LDLT for a patient with hepatitis B-related cirrhosis utilizing a graft from an HBsAg positive 'healthy carrier' donor using a combination prophylaxis of lamivudine and adefovir dipivoxil. RESULTS: To date, the patient has been doing well with normal liver function tests and liver histological findings at 4 years after the transplantation and the donor has also been doing well. CONCLUSIONS: Although virological recurrence appears to be universal despite prophylaxis, re-evaluation of the use of a graft from a healthy HBsAg-positive donor is warranted in this era of combination prophylaxis. PMID- 17919242 TI - Branched-chain amino acids improve insulin resistance in patients with hepatitis C virus-related liver disease: report of two cases. AB - Hepatitis C virus (HCV) infection causes insulin resistance. Because increased insulin resistance is a risk factor for development of hepatocellular carcinoma and reduced long-term survival, insulin resistance is a therapeutic target in patients with HCV infection. Branched-chain amino acids (BCAAs) are not only structural constituents of proteins but they are also considered as regulators of insulin signalling. We first describe two cases suggesting that administration of BCAAs improves insulin resistance associated with HCV-related liver disease. Although there were no changes in body weight, plasma glucose concentration and haemoglobin A1c (HbA1c) value were decreased. Moreover, BCAAs caused a decrease in both fasting insulin concentration and the value of homeostasis model assessment for insulin resistance. Thus, BCAAs are a potential therapeutic agent for improving insulin resistance in patients with HCV-related liver disease. PMID- 17919240 TI - NIM811, a nonimmunosuppressive cyclosporine analogue, suppresses collagen production and enhances collagenase activity in hepatic stellate cells. AB - BACKGROUND/AIMS: A recent decrease in patient survival has been reported among hepatitis C virus (HCV)-infected liver transplant recipients and this may be attributable to progression of fibrosis. We reported previously that cyclosporine suppressed the proliferation of, and collagen production in, hepatic stellate cells (HSCs). Here, we investigated the effects of NIM811, a cyclosporine analogue, on cell growth, collagen production and collagenase activity in HSCs. METHODS: Rat HSCs and human HSC-derived TWNT-4 cells were cultured for the study. The expression of collagen, matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) and collagenase activity was evaluated. Cell proliferation and apoptosis were measured. Phosphorylation of mitogen activated protein kinases (MAPKs), Smad2 and Smad3 was evaluated. The expression of the tumour growth factor-beta (TGF-beta)-receptor and Smad7 genes was also evaluated. RESULTS: NIM811, as well as cyclosporine, suppressed the transcription and synthesis of collagen and stimulated the production of MMP-1 with a concomitant enhancement of collagenase activity, although it did not change the expression of TIMP-1. NIM811 inhibited proliferation without induction of apoptosis. In the MAPKs and TGF-beta signalling pathways, NIM811 enhanced the phosphorylation of JNK and p38, but not extracellular signal-regulated kinases 1 and 2, and suppressed the phosphorylation of Smad2 and Smad3, accompanied by increased Smad7 transcription and decreased TGF-beta-receptor transcription. CONCLUSION: These findings demonstrate that NIM811 not only suppresses collagen production and proliferation but also increases collagenase activity. These effects are accompanied by inhibition of TGF-beta signalling pathways. PMID- 17919243 TI - Perivascular epithelioid cell tumour of the liver. PMID- 17919244 TI - Association between food mixing ability and mandibular movements during chewing of a wax cube. AB - The purpose of this study was to clarify whether masticatory movements during chewing of a wax cube associate with food mixing ability. Twenty-six dentate subjects (mean age 25.3 years) were directed to chew a two-coloured paraffin wax cube for 10 strokes on preferred chewing side. Mixing Ability Index (MAI) was determined from the colour mixture and shape of the chewed wax cube. MAI was employed as an estimate of food mixing ability. Mandibular movements during chewing of the wax cube were recorded using a six-degrees-of freedom jaw movements recording system, and motion of a lower incisal point of the mandible was computed. Twelve parameters of masticatory movements in relation to mandibular excursion, angle, velocity, cycle duration, duration of each phase were measured for each masticatory cycle. In addition, unilateral maximum biting was performed, and the maximum force during biting was measured using a force transducer. A multiple regression analysis identified vertical amplitude, closing duration, closing angle and maximum bite force as significant predictors accounting for 63% of inter-subject variation (adjusted R(2)) in the MAI (P < 0.05). Only some parameters of masticatory movements seem to have an influence on the MAI in dentate subjects. PMID- 17919245 TI - Correlations between incisor and condylar movements during lateral excursion in children with primary dentition. AB - The purpose of this study was to look for associations between lower incisor movement and working and balancing condylar movement during lateral excursion in children with primary dentition and adults with permanent dentition. Mandibular movement was recorded using a TRI-MET (an optoelectronic analysis system with six degrees-of-freedom) at a sampling frequency of 100 Hz. The movement data was transferred to a graphics workstation for analysis. Subjects were categorized by age into two groups. The primary dentition group consisted of 19 children (mean age: 5 years and 5 months, s.d.: 8.7 months), and the permanent dentition group consisted of 22 women (mean age: 20 years and 5 months, s.d.: 26.3 months). The occlusion and TMJ in both groups were normal, with no history of orthodontic treatment. Three orthogonal excursive ranges and the 3D linear distance of the incisal and balancing and working condylar points, along with inter- and intra individual correlations between the incisor and the balancing and working condyles during lateral excursion, were estimated by using multilevel statistical models. Lateral excursion in children with primary dentition was characterized by smaller incisor excursive ranges and 3D linear distance than in adults, and stronger inter- and intra-individual correlations between incisor and balancing condylar movements than in adults. In both children and adults the lateral excursion of the incisor was a good indicator of the extent of balancing condylar movement, but not working condylar movement. PMID- 17919246 TI - Short-term efficacy of physical therapy compared to splint therapy in treatment of arthrogenous TMD. AB - A prospective randomized study was carried out to evaluate the efficacy of physical therapy in addition to splint therapy on treatment outcome in patients with temporomandibular disorders (TMD) with respect to objective and subjective parameters. Twenty-six patients suffering from an arthrogenic TMD and exhibiting a painfully restricted jaw opening were randomized in two groups. Thirteen patients were treated solely with Michigan splint (group I), 13 patients received supplementary physical therapy (group II). Before treatment a clinical examination and electronic recording of jaw movements were performed and subjective pain level was evaluated by visual analogue scales. After 3 months of therapy maintenance of improvement was evaluated. Within treatment groups comparison of data before and after treatment was analysed using Wilcoxon test. Groups were compared by Mann-Withney-U test. A P-value < 0.05 was considered significant. Compared with the baseline, in both groups mandibular movement capacity increased significantly after treatment, whereas subjective pain decreased significantly (P < 0.05). Active jaw opening increased from 28.6 +/- 5.8 to 35.9 +/- 4.8 mm in group I and from 30.1 +/- 5.4 to 40.8 +/- 4.1 mm in group II. After therapy the difference of active jaw opening between groups was significant (P < 0.05). Physical therapy also gave a supplementary improvement of protrusive mandibular movement capacity during electronic registration and subjective pain level. For none of these parameters this difference between groups was significant. Physical therapy seems to have a positive effect on treatment outcome of patients with TMD. PMID- 17919247 TI - Do tooth gaps matter? Evaluation of self-assessments: a pilot study. AB - This study aimed to test the hypothesis stating no difference in number of teeth or tooth gaps counted by the dentist or the patient. The sample consisted of 49 randomly selected regular patients and their dentists (response rate 10%, 53% males). Mean age of the patients was 53.6 +/- 11.9 years. The mean number of teeth was 26.5, 20/49 reported tooth gaps (excluding second and third molars) and 12/20 had visible gaps. The calculation of patients' detection of gaps showed a sensitivity = 79%, specificity = 96% and a positive predictive value of 95%. The patients reported more teeth in the maxilla and fewer teeth in the mandible compared with their dentists. More teeth were misdiagnosed as the number of teeth increased in the mandible. Patients reported more gaps than dentists in both jaws (NS); an average of 0.04, 0.18 and 0.22 misdiagnosed gaps in the maxilla, mandible and for both jaws respectively. Agreement on tooth present/missing in the maxilla was 91.3% (+/-9.82) and in the mandible 88.2% (+/-10.44). Correlation between self-report and dentist's report of the number of teeth present and the number of gaps in the maxilla was (Pearson's r) 0.94 and 0.83. One person with self-perceived visible gaps and five persons with non-visible gaps were recommended treatment; two persons agreed about a treatment need. Self assessments of tooth gaps show limited but acceptable sensitivity and excellent specificity. No significant differences in tooth counting were found when comparing self-reports and dentists' reports. The study should be tried on a larger scale. PMID- 17919248 TI - Reliability and validation of a Brazilian version of the Oral Health Impact Profile for assessing edentulous subjects. AB - The aims of this study were to evaluate the reliability and to validate a Brazilian version of Oral Health Impact Profile for assessing edentulous subjects (OHIP-EDENT), an inventory for measuring oral health-related quality of life of edentulous subjects. The sample comprised 65 complete denture wearers (23 men, mean age of 69.1 +/- 10.3 years). The translated OHIP-EDENT was applied on two occasions with a washout period of 3 months. Reliability was assessed by an internal consistency analysis and a test-retest approach. A preliminary validation process was conducted by a qualitative approach/interview. Results of internal consistency showed a Cronbach's alpha of 0.86 or 0.90 for the first or second appointment respectively. Through the test-retest analysis, an intra-class correlation coefficient of 0.57 was found, and individual answers reflected a broad range of agreement. Interviewed volunteers (n = 6) comprehended most questions well. In conclusion, the Brazilian version of OHIP-EDENT is adequate for assessing the oral health-related quality of life for edentulous subjects. PMID- 17919249 TI - Influence of tongue pressure and width on tongue indentation formation. AB - The purpose of this study was to clarify which physiological and anatomical factors were involved in the formation of tongue indentations, which are believed to be a clinical sign of clenching. Twenty-four dentulous subjects were investigated. They were divided into two groups, depending on the presence or absence of tongue indentations: (i) a tongue indentation group and (ii) a no tongue indentation group. Intraoral appliances containing a small pressure sensor were placed at the lingual surfaces of the upper and lower right first molars. Lingual pressure on the lingual surfaces of the upper and lower right first molars was then recorded under different conditions. The tasks selected as physiological factors to be recorded were: (i) silent reading at rest for 10 min, (ii) maximum voluntary clenching (MVC) for 5 s, (iii) 10% of MVC for 1 min and (iv) swallowing. The results for all tasks were compared between groups. Tongue width and dental arch width were also measured as anatomical factors. No significant differences were found between groups during silent reading at rest, clenching, swallowing, or in tongue pressure integration during silent reading at rest. However, a significant difference was found in terms of tongue width (P < 0.05). These results indicate that neither volume of lingual pressure exerted on the lingual surface of the teeth during rest, clenching, nor swallowing are related to the formation of tongue indentations. The results do suggest; however, that tongue width at rest plays an important role in the formation of tongue indentations. PMID- 17919250 TI - A preliminary report of patients treated with early loaded implants using computerized tomography-guided surgical stents: flapless versus conventional flapped surgery. AB - The objective of this clinical study was to compare the survival rates of early loaded implants placed using flapless and flapped surgical techniques and to determine the bone density in the implant recipient sites using computerized tomography (CT). The study population consisted of 12 patients who were referred implant placement. One group consisted of five patients referred for the placement of 14 implants and treated with a flapless procedure. The other group consisted of seven patients referred for the placement of 45 implants with a conventional flapped procedure. Patients were selected randomly. CT machine was used for pre-operative evaluation of the jaw bone and the mean bone density value of each implant recipient site was recorded in Hounsfield units (HU). All implants were placed using CT-guided surgical stents. The early loading protocols included 2 months of healing in the mandible and 3 months of healing in the maxilla. Single-implant crowns, implant-supported fixed partial dentures, and implant-retained over dentures were delivered to the patients. Of 59 implants placed, one was lost in the conventional flapped group within the first month of healing, meaning overall implant survival rate of 98.3% average 9 months later. The highest average bone density value (801 +/- 239 HU) was found in the anterior mandible, followed by 673 +/- 449 HU for the posterior maxilla, 669 +/-346 HU for the anterior maxilla and 538 +/- 271 HU for the posterior mandible. The results of this study show that the early loading of implants placed utilizing flapless surgical technique with CT-guided surgical stents may be possible. PMID- 17919251 TI - Influence of thermal expansion mismatch and fatigue loading on phase changes in porcelain veneered Y-TZP zirconia discs. AB - There is a rapidly growing interest for the use of Y-TZP zirconia as core material in veneered all-ceramic prostheses. It was hypothesized that a mismatch in coefficient of thermal expansion between the veneering porcelain and the Y-TZP zirconia core of these prostheses causes transformation of the tetragonal to the monoclinic structure in Y-TZP zirconia at the interface boundary when exposed to fatigue loading, resulting in fracture at the interface boundary. Y-TZP zirconia discs were veneered with three porcelains differing in coefficient of thermal expansion. Finite element analysis was used to investigate the stress distribution in the bi-layered discs because of the mismatch in thermal expansion. Two of these three groups were fatigued with the veneering ceramic in tension. X-ray diffraction was used to measure the intensity of monoclinic and tetragonal zirconia phase present at the zirconia core surface after sintering, airborne abrasion, veneering, and fatigue loading. It was found that the sintered tetragonal structure was converted to monoclinic up to a depth of 27 mum after airborne abrasion, and reversed back to tetragonal after porcelain veneering with porcelain. Fatigue loading of veneered discs did not, even with the highest possible thermal mismatch stress, cause any conversion from tetragonal to monoclinic phase. PMID- 17919252 TI - Comparison of shade matching by visual observation and an intraoral dental colorimeter. AB - The purpose of this study was to compare the applicability of two shade-matching approaches: Vintage Halo shade guide (visual method) and Shofu ShadeEye NCC colorimeter (instrumental method). Twenty participants' maxillary left central incisors were evaluated. Corresponding metal ceramic crowns were fabricated with each shade-matching approach. The colour distributions (L*, a* and b*) of the middle third region of each tooth and corresponding metal ceramic crowns were spectrophotometrically assessed. The colour difference (DeltaE) and colour distributions (DeltaL*, Deltaa* and Deltab*) between the tooth and the corresponding crowns were calculated. We found that the colour differences of both groups fell within the clinical unacceptable range (DeltaE > 2.75). Regarding DeltaE and the three colour distributions, no significant difference was found, expect for a* (P < 0.01). The shade matching difficulty degree was analysed through the agreements of visual shade selection. Within easy matching cases, the instrumental method achieved better results (P = 0.041). In conclusion, it is suggested that the reliability of shade matching can be ensured by neither the colorimeter nor the visual approach. However, the colorimeter can achieve better results within easy matching cases. PMID- 17919253 TI - Clinical performance of a self-etching and a total-etch adhesive system - 2-year results. AB - The purpose of this controlled prospective clinical study was to compare the clinical performance of the self-etching adhesive system AdheSE and the established total-etch adhesive system Excite in classes I and II cavities for a period of 2 years. Fifty patients participated in this study. Each received two restorations, one with AdheSE, one with Excite by one calibrated, non-blinded operator. The resin composite used to restore the teeth was Tetric Ceram HB. One calibrated, blinded clinician re-evaluated the restorations at baseline, after 6, 12 and 24 months after placement using the modified Ryge criteria. For this, vitality, post-operative sensitivity, visible marginal irregularity, marginal discolouration, secondary caries, surface texture, anatomic form and filling defect were considered. After 2 years, 67 restorations were reviewed in 34 patients. None of the teeth showed signs of secondary caries. Two teeth, one of each group, had to receive endodontic treatment because of pulp inflammation. All other teeth remained vital. Slight marginal discolourations were observed in six AdheSE restorations and three Excite restorations. These restorations were scored as Beta. After 2 years, an overall clinical success rate, summing up all the Alpha and Beta scores, of 97% was found, viewing both adhesive systems together. Statistic analysis using log-rank test showed no statistic differences in the overall survival rate between the two materials tested within the observation period. It can be concluded that both adhesive systems tested demonstrated very good clinical performance in the restoration of classes I and II cavities at the end of 2 years. PMID- 17919254 TI - Clinical effects of acrylic resilient denture liners applied to mandibular complete dentures on the alveolar ridge. AB - The purpose of this study was to investigate whether resilient denture liners has the potential to decrease problems affecting the alveolar ridge mucosa during the first adjustment session following the initial fitting of the denture. Interventions were complete denture treatment with permanent acrylic resilient denture liner dentures (RLD) in mandibles and conventional heat-activated acrylic resin dentures (ARD) in mandibles. Maxillary dentures were fabricated with conventional heat-activated ARD. Seventy-four subjects were randomly allocated to the RLD and ARD groups by random permuted blocks within the strata method after obtaining written informed consent. A randomized controlled parallel clinical trial with two hospital centres was conducted from April 2004 to July 2006. The problems were comprehensively evaluated by three outcomes with subjective and objective factors as follows: (i) numbers of sore spots, (ii) duration of wearing dentures and (iii) pain perception with 100 mm Visual Analog Scale. The numbers of sore spots in the mandibular support and border areas for the RLD group were significantly less than those for the ARD groups (P < 0.05). The RLD wearers were able to wear their dentures an additional day longer than the ARD wearers (P < 0.05). The maxillary and mandibular pain ratings for the RLD group were significantly lower than those for the ARD group (P < 0.05). Application of resilient denture liners to mandibular complete dentures provided edentulous patients with fewer problems affecting the alveolar during the first adjustment session following the initial fitting of the denture when compared with conventional denture treatments. PMID- 17919256 TI - F90927: a new member in the class of cardioactive steroids. AB - F90927 is a newly developed cardioactive drug with a steroid-like structure. It acts directly and agonistically on the cardiac L-type Ca2+ channel by shifting its voltage-dependent activation toward more negative potentials. This leads to an increased influx of Ca2+ and, therefore, to a stronger contraction; however, no arrhythmias occur. Calcium current stimulation can already be observed at nanomolar concentrations, but higher concentrations of F90927 elevate intracellular Ca2+ concentration, causing a reduction of the myocardial compliance and an increased diastolic blood pressure. Vessels also react to F90927 and contract in its presence. Binding of F90927 with the L-type Ca2+ channel presumably occurs in the vicinity of the transmembrane domains III and IV of the alpha1 subunit. F90927 exhibits no use dependence and interacts with Ca2+ channel inhibitors of all three known classes of channel modulators (dihydropyridines, phenylalkylamines, and benzothiazepines), suggesting that it is a member of a new class of Ca2+ channel modulators. Due to its adverse effects on blood pressure and vessel contraction, F90927 is not an ideal drug candidate. It has, however, some unique properties, which makes it a promising tool to study the function of the L-type Ca2+ channel. PMID- 17919257 TI - Edifoligide: a transcription factor decoy to modulate smooth muscle cell proliferation in vein bypass. AB - The era of genomics and recombinant DNA technology has ushered in an entirely new class of therapeutic agents designed to influence disease progression at a genetic level. The scope and utility of this technology is not fully realized. However, multiple trials of therapeutic agents have been completed and many more are ongoing. Here we report on edifoligide, a double-stranded oligodeoxynucleotide (ODN) that competitively inhibits the transcription factor E2F, a critical regulator of the cell cycle. Edifoligide has undergone extensive clinical testing for the treatment of intimal hyperplasia following vascular bypass procedures. In this review we address the rationale for targeting E2F in vascular disease, the pharmacology of edifoligide, and the results of preclinical and clinical studies using this novel compound. PMID- 17919259 TI - Conivaptan: a dual vasopressin receptor v1a/v2 antagonist [corrected]. AB - Several fluid retentive states such as heart failure, cirrhosis of the liver, and syndrome of inappropriate antidiuretic hormone secretion are associated with inappropriate elevation in plasma levels of arginine vasopressin (AVP), a neuropeptide that is secreted by the hypothalamus and plays a critical role in the regulation of serum osmolality and in circulatory homeostasis. The actions of AVP are mediated by three receptor subtypes V1a, V2, and V1b. The V1a receptor regulates vasodilation and cellular hypertrophy while the V2 receptor regulates free water excretion. The V1b receptor regulates adrenocorticotropin hormone release. Conivaptan is a nonpeptide dual V1a/V2 AVP receptor antagonist. It binds with high affinity, competitively, and reversibly to the V1a/V2 receptor subtypes; its antagonistic effect is concentration dependent. It inhibits CYP3A4 liver enzyme and elevates plasma levels of other drugs metabolized by this enzyme. It is approved only for short-term intravenous use. Infusion site reaction is the most common reason for discontinuation of the drug. In animals conivaptan increased urine volume and free water clearance. In heart failure models it improved hemodynamic parameters and free water excretion. Conivaptan has been shown to correct hyponatremia in euvolemic or hypervolemic patients. Its efficacy and safety for short-term use have led to the Food and Drug Administration (FDA) approval of its intravenous form for the correction of hyponatremia in euvolemic and hypervolemic states. Despite its ability to block the action of AVP on V1a receptors, no demonstrable benefit from this action was noted in patients with chronic compensated heart failure and it is not approved for this indication. Consideration should be given to further evaluation of its potential benefits in patients with acute decompensated heart failure. PMID- 17919260 TI - The functional consequence of the Glu298Asp polymorphism of the endothelial nitric oxide synthase gene in young healthy volunteers. AB - OBJECTIVE: To assess the role of the endothelial nitric oxide synthase (eNOS) gene variant as a risk factor for atherosclerosis we sought to investigate whether the Glu298Asp polymorphism of the eNOS gene is associated with functional changes in the endothelium in healthy volunteers. METHODS: Endothelial function was assessed in 68 normal volunteers (ages 18-44 years) by bilateral forearm venous occlusion plethysmography with intraarterial infusions of increasing doses of acetylcholine for endothelial-dependent vasodilation and, with sodium nitroprusside and verapamil for endothelial-independent vasodilation. Blood was genotyped by polymerase chain reaction and BanII digestion. RESULTS: Asp homozygotes (TT) had a decreased vasodilatory response to acetylcholine [forearm blood flow (FBF) ratio between infused and control arm, 2.82 +/- 1.10] when compared to GG variant (FBF ratio to acetylcholine, 3.97 +/- 1.90, p= 0.04) and to a certain extent, the GT variant (FBF ratio to acetylcholine, 3.79 +/- 2.28, p= 0.07). There was no effect of eNOS genotype on the response to the endothelial independent vasodilators-sodium nitroprusside and verapamil. CONCLUSIONS: Our data show that carriage of the Asp298 variant of the eNOS gene is associated with a blunted endothelial-dependent vasodilation in healthy volunteers. These findings support a genetically determined modulation of endothelial dysfunction, a phenotype of early atherosclerosis in humans. PMID- 17919263 TI - Abstracts of Australian Gastroenterology Week. October 24-27, 2007. Perth, Australia. PMID- 17919258 TI - Role of poly(ADP-ribose) polymerase 1 (PARP-1) in cardiovascular diseases: the therapeutic potential of PARP inhibitors. AB - Accumulating evidence suggests that the reactive oxygen and nitrogen species are generated in cardiomyocytes and endothelial cells during myocardial ischemia/reperfusion injury, various forms of heart failure or cardiomyopathies, circulatory shock, cardiovascular aging, diabetic complications, myocardial hypertrophy, atherosclerosis, and vascular remodeling following injury. These reactive species induce oxidative DNA damage and consequent activation of the nuclear enzyme poly(ADP-ribose) polymerase 1 (PARP-1), the most abundant isoform of the PARP enzyme family. PARP overactivation, on the one hand, depletes its substrate, NAD+, slowing the rate of glycolysis, electron transport, and ATP formation, eventually leading to the functional impairment or death of the endothelial cells and cardiomyocytes. On the other hand, PARP activation modulates important inflammatory pathways, and PARP-1 activity can also be modulated by several endogenous factors such as various kinases, purines, vitamin D, thyroid hormones, polyamines, and estrogens, just to mention a few. Recent studies have demonstrated that pharmacological inhibition of PARP provides significant benefits in animal models of cardiovascular disorders, and novel PARP inhibitors have entered clinical development for various cardiovascular indications. Because PARP inhibitors can enhance the effect of anticancer drugs and decrease angiogenesis, their therapeutic potential is also being explored for cancer treatment. This review discusses the therapeutic effects of PARP inhibitors in myocardial ischemia/reperfusion injury, various forms of heart failure, cardiomyopathies, circulatory shock, cardiovascular aging, diabetic cardiovascular complications, myocardial hypertrophy, atherosclerosis, vascular remodeling following injury, angiogenesis, and also summarizes our knowledge obtained from the use of PARP-1 knockout mice in the various preclinical models of cardiovascular diseases. PMID- 17919265 TI - Polymorphism of interleukin-18 promoter influences the onset of kidney graft function after transplantation. AB - It has been well recognized that the promoter polymorphisms of interleukin-18 (IL 18) influence the level of cytokine expression. In our previously published data, we showed constitutive IL-18 expression in the epithelium of renal distal tubules in patients after kidney transplantation and significantly elevated IL-18 expression during acute rejection. In this study, we evaluated the clinical significance of two functional promoter polymorphisms of the IL-18 gene at positions -607 A/C (rs1946518) and -137 C/G (rs187238) in patients after kidney transplantation and looked for associations with the onset of graft function and the incidence of rejection episodes. Promoter polymorphisms in 124 patients and 103 unrelated controls were evaluated by sequence-specific primer polymerase chain reaction and the allele, genotype and haplotype frequencies were statistically correlated. We found a statistically different distribution of the allele frequency of -607 A/C polymorphism between patients with immediate or delayed onset of kidney graft function. Data showed that the C allele, which contributes to higher IL-18 expression, is more frequent in patients with delayed onset of function (P = 0.03, odds ratio = 1.93; 95% confidence interval = 1.15 3.25). A/C single nucleotide polymorphisms of the IL-18 promoter at position -607 may influence the onset of early kidney allograft function. PMID- 17919264 TI - Characterization of mouse CD229 (Ly9), a leukocyte cell surface molecule of the CD150 (SLAM) family. AB - Mouse CD229 (Ly9) is a cell surface molecule of the CD150 (signaling lymphocyte activation molecule) family. This family consists of nine leukocyte receptors of the immunoglobulin superfamily that are involved in leukocyte activation. CD229 binds to SAP, a protein encoded by the gene for X-linked lymphoproliferative disease. In this study, mouse CD229 expression was assessed with a new CD229 specific monoclonal antibody (mAb) (Ly9.ab3), raised using CD229-transfected cells. CD229 was expressed on Sca-1+c-kit+Lin- hematopoietic stem cells, and this expression increased during lymphocyte maturation. Virtually, all T and B cells expressed high levels of CD229. CD229 was absent on granulocytes, bone marrow derived dendritic cells, platelets, and red blood cells (RBCs). However, it was expressed at significant levels on monocytes, indicating that it is also expressed on mouse myeloid cells. We also show that natural killer cells, natural killer T cells, and B1 cells express very high levels of this molecule. In vitro functional experiments showed that ligation of CD229 inhibited the expression of the activation markers CD69 and CD25 on T lymphocytes in response to anti-CD3 stimulation. Moreover, this reduced activation was concurrent with a reduction in cytokine production. Our results show that CD229 is a pan-lymphocyte marker and indicate that mAbs against CD229 are able to down-modulate T-cell activation. PMID- 17919267 TI - Sequence analysis of the novel HLA-B35 (B*3576) allele in an African individual. AB - A novel human leucocyte antigen (HLA)-B35 (HLA-B*3576) allele has been described in an African individual by polymerase chain reaction sequence-based typing. This new allele contains six nucleotide substitutions and is homologous to B*3501 with the exception of residues 66-74 resulting in five amino acid mutations. PMID- 17919268 TI - Nomenclature for factors of the HLA system, update July 2007. PMID- 17919266 TI - Islet autoantibodies are associated with HLA-DQ genotypes in Han Chinese patients with type 1 diabetes and their relatives. AB - The objective of this study was to explore the relationship between islet autoantibodies of glutamic acid decarboxylase (GADA), islet antigen-2A (IA-2A), insulin autoantibody (IAA), and human leukocyte antigen (HLA)-DQ genotypes in type 1 diabetes (T1D) patients and their first-degree relatives (FDRs). Cross sectional and case-control study. Four hundred and ninety-five T1D patients, 419 FDRs, and 376 control subjects in Han Chinese populations were recruited and tested for GADA and IA-2A, while 71 cases, all FDRs and 300 controls were tested for IAA. The 338 T1D patients (including 187 antibody-positive and 151 antibody negative patients), 173 FDRs and 278 controls were genotyped for HLA-DQ with polymerase chain reaction sequencing-based method. Compared with the control, the frequency of DQA1*03-DQB1*0303, DQA1*05-DQB1*0201, and DQA1*03-DQB1*0401 haplotypes was higher (P < 0.05-0.01) but DQA1*0102-DQB1*0602 haplotype was lower (P < 0.01) in T1D patients. DQA1*03 allele was less in the FDRs than in their probands (P < 0.05). GADA was more prevalent in T1D patients carrying DQA1*05 DQB1*0201 or DQA1*03-DQB1*0401 haplotype (55.8% vs 41.0%, 65.5% vs 40.3%, P < 0.05-0.01), whereas IA-2A presented more in the patients carrying DQA1*03 DQB1*0303 haplotype (27.0% vs 7.9%, P < 0.05-0.01), both GADA and IA-2A showed frequently in the patients with DQA1*03-DQB1*0303/DQA1*05-DQB1*0201 haplotypes (34.5% vs 9.7%, P < 0.01). GADA positivity was lower in the patients with DQA1*0102-DQB1*0602 haplotype (16.7% vs 45.9%, P < 0.05). The frequency of IAA was not different between patients with and without susceptible DQ haplotypes (P > 0.05). GADA, IA-2A or IAA presented frequently in FDRs with DQA1*03-DQB1*0303 haplotype. The findings in the study indicate that some of specific HLA-DQA1/ DQB1 genotypes and haplotypes not only confer susceptibility to T1D but also are associated with the presence of the islet autoantibodies in the Han Chinese population. PMID- 17919270 TI - Clinical trial: multispecies probiotic supplementation alleviates the symptoms of irritable bowel syndrome and stabilizes intestinal microbiota. AB - BACKGROUND: Irritable bowel syndrome is the most common diagnosis in gastroenterology. Trials suggest certain probiotics to be beneficial. AIM: To investigate the effects of multispecies probiotic supplementation (Lactobacillus rhamnosus GG, L. rhamnosus Lc705, Propionibacterium freudenreichii ssp. shermanii JS and Bifidobacterium animalis ssp. lactis Bb12) on abdominal symptoms, quality of life, intestinal microbiota and inflammatory markers in irritable bowel syndrome. METHODS: Eighty-six irritable bowel syndrome patients (Rome II criteria) participated in this randomized, placebo-controlled 5-month intervention. Patients were randomized to receive daily either multispecies probiotic supplementation or placebo. Irritable bowel syndrome symptoms, quality of life, microarray-based intestinal microbiota stability (n = 20), serum cytokines and sensitive C-reactive protein were monitored. RESULTS: The composite irritable bowel syndrome score had at 5 months decreased 14 points (95% CI: -19 to -9) from baseline with the multispecies probiotic vs. three points (95% CI: -8 to 1) with placebo (P = 0.0083). Especially, distension and abdominal pain were affected. A stabilization of the microbiota was observed, as the microbiota similarity index increased with the probiotic supplementation (1.9 +/- 3.1), while it decreased with placebo (-2.9 +/- 1.7). No differences were seen in C reactive protein. CONCLUSIONS: This multispecies probiotic seems to be an effective and safe option to alleviate symptoms of irritable bowel syndrome, and to stabilize the intestinal microbiota. PMID- 17919271 TI - Cumulative incidence of chronic constipation: a population-based study 1988-2003. AB - AIM: To estimate the cumulative incidence of chronic constipation and evaluate potential risk factors. METHODS: In previous cross-sectional studies in 1988, random samples of Olmsted County, MN residents were mailed valid gastrointestinal symptoms surveys. A similar survey was mailed in 2003 to all the remaining eligible subjects who had been mailed to previously. An incident case of chronic constipation was defined as no reported constipation or irritable bowel syndrome on their initial survey but reported constipation on the second survey. RESULTS: In all, 5507 (79%) subjects responded to the initial survey and 2298 (55%) subjects responded to the second survey in which chronic constipation could be defined. Over 12 years, the cumulative incidence of chronic constipation was 17.4% (14.5, 20.5). Among those less than age 50 years at baseline, the incidence of chronic constipation differed by gender (9.2% in men vs. 18.3% in women). In those over 70 years, the incidence of chronic constipation was more similar for men and women (20.6% vs. 25.0%). The other risk factor associated with new onset chronic constipation was the presence of abdominal pain at baseline [OR = 2.0 (1.3, 3.0)]. CONCLUSION: The cumulative incidence of chronic constipation over more than a decade was almost one in six, and more pronounced in women and the elderly. PMID- 17919273 TI - Metabolic syndrome is associated with severe fibrosis in chronic viral hepatitis and non-alcoholic steatohepatitis. AB - BACKGROUND: The prevalence of metabolic syndrome and its possible impact on the severity of liver histological lesions have not been studied prospectively in chronic liver diseases. AIM: To investigate the prevalence of metabolic syndrome in patients with chronic viral hepatitis or non-alcoholic steatohepatitis, and to determine its associations with histological severity. METHODS: We prospectively included 317 patients (hepatitis B e antigen-negative chronic hepatitis B: 95, chronic hepatitis C: 176, non-alcoholic steatohepatitis: 46) with liver biopsy. Metabolic syndrome was defined using the Adult Treatment Panel III criteria. Histological lesions were evaluated according to Ishak's or Brunt's classification. RESULTS: Metabolic syndrome was present in 10.4% of patients being significantly more prevalent in non-alcoholic steatohepatitis than in chronic viral hepatitis (41.3% vs. 5.1%, P < 0.001). In chronic viral hepatitis, cirrhosis (stages 5-6) was independently associated with increasing age, higher aspartate aminotransferase and gamma-glutamyl-transpeptidase levels, severe necroinflammation and metabolic syndrome (P = 0.016). In non-alcoholic steatohepatitis, severe fibrosis (stages 3-4) was independently associated with severe necroinflammation and metabolic syndrome (P = 0.033). Presence of metabolic syndrome was not associated with presence or severity of steatosis both in chronic viral hepatitis and in non-alcoholic steatohepatitis. CONCLUSION: Metabolic syndrome is more prevalent in non-alcoholic steatohepatitis than in chronic viral hepatitis; it is associated independently with more severe fibrosis but not with the severity of steatosis, both in chronic viral hepatitis and in non-alcoholic steatohepatitis. PMID- 17919272 TI - Clinical trial: The effect of somatostatin vs. octreotide in preventing post endoscopic increase in hepatic venous pressure gradient in cirrhotics with bleeding varices. AB - BACKGROUND: Hepatic venous pressure gradient (HVPG) increases significantly after endoscopic therapy in patients with bleeding oesophageal varices, which may precipitate further haemorrhage. Whether vasoactive drugs can suppress these changes remains unknown. AIM: To investigate the efficacy of somatostatin when compared with octreotide in preventing the post-endoscopic increase in HVPG during acute bleeding and whether the changes affect outcome. METHODS: Thirty three cirrhotics with bleeding varices were randomized to receive somatostatin (n = 17) or octreotide (n = 16) under double-blind conditions, soon after their admission. HVPG measurements were performed before and immediately after endoscopic treatment. RESULTS: In the somatostatin group, postendotherapy HVPG values did not change significantly when compared with pre-treatment values (18.9 vs. 17.2, P = 0.092). Conversely, in the octreotide group, HVPG increased significantly after endoscopy (18.2 vs. 20.8, P = 0.003). The probability of 6 week survival without treatment failure was significantly higher in the somatostatin group (P = 0.024). Post-endoscopic HVPG value was independently associated with 6-week failure. CONCLUSIONS: Somatostatin, but not octreotide, effectively prevents the post-endoscopic increase in HVPG, which may be associated with low probability of treatment failure. PMID- 17919274 TI - Severe symptoms of gastro-oesophageal reflux disease are associated with cardiovascular disease and other gastrointestinal symptoms, but not diabetes: a population-based study. AB - BACKGROUND: Few population-based studies have examined comorbidity in relation to gastro-oesophageal reflux disease (GERD). AIM: To study the association between cardiovascular disease, diabetes, gastrointestinal symptoms and GERD. METHODS: Population-based, cross-sectional, case-control study based on a large Norwegian health survey conducted in 1995-97. Among 65,333 participants, 3153 persons reporting severe reflux symptoms were defined as cases, and 40,210 persons without such symptoms were defined as controls. Data on cardiovascular disease, diabetes, gastrointestinal symptoms and potential confounders were collected through questionnaires. Odds ratios (OR) with 95% confidence intervals (CI) were estimated using unconditional logistic regression, in crude and adjusted models. RESULTS: In the crude models, positive associations were observed between myocardial infarction (OR 1.7, 95% CI 1.4-2.1), angina pectoris (OR 2.5, 95% CI 2.1-2.9) and stroke (OR 1.6, 95% CI 1.2-2.1) and risk of GERD. The associations were attenuated in the adjusted models, but remained significant for angina pectoris (OR 1.9, 95% CI 1.6-2.2). No association was observed between diabetes and GERD. Strong positive associations were seen between all studied gastrointestinal symptoms, i.e. nausea, diarrhoea and constipation, and risk of GERD. CONCLUSIONS: This population-based study indicates that myocardial infarction, angina pectoris, stroke and symptoms of nausea, diarrhoea and constipation are associated with GERD. PMID- 17919275 TI - Review article: the evidence base for interventions used to maintain remission in Crohn's disease. AB - BACKGROUND: Crohn's disease is characterised by recurrent flare-ups alternating with periods of remission. A number of interventions are currently used in clinical practice to try and maintain remission in Crohn's disease but the evidence base for some of them may be questionable. AIM: To review the available evidence on interventions, which are currently used to maintain remission in Crohn's disease. METHODS: The Cochrane Library and Medline (Pubmed) were searched for level 1 evidence on specific interventions. Search terms included 'Crohn's disease or synonyms', 'remission or synonyms' and the names of specific interventions. RESULTS: Azathioprine, infliximab and adalimumab are effective at maintaining remission in Crohn's disease. Natalizumab is also effective, but there are concerns about its potential association with progressive multifocal leukoencephalopathy. Long-term enteral nutritional supplementation, enteric coated omega-3 fatty acids and intramuscular methotrexate may also be effective but the evidence for these is based on relatively small studies. The available evidence does not support the use of oral 5-aminosalicylates agents, corticosteroids, anti-mycobacterial agents, probiotics or ciclosporin as maintenance therapy in Crohn's disease. CONCLUSION: A better understanding of the evidence base of existing interventions could result in the use of treatments, which are more likely to lead to improved patient outcomes. PMID- 17919276 TI - Oral pathology in untreated coeliac [corrected] disease. AB - BACKGROUND: Many coeliac disease patients with atypical symptoms remain undiagnosed. AIM: To examine the frequency of oral lesions in coeliac disease patients and to assess their usefulness in making coeliac disease diagnosis. PATIENTS AND METHODS: One hundred and ninety-seven coeliac disease patients and 413 controls were recruited and the oral examination was performed. RESULTS: Forty-six out of 197 coeliac disease patients (23%) were found to have enamel defects vs. 9% in controls (P < 0.0001). Clinical delayed eruption was observed in 26% of the pediatric coeliac disease patients vs. 7% of the controls (P < 0.0001). The prevalence of oral soft tissues lesions was 42% in the coeliac disease patients and 2% in controls (P < 0.0001). Recurrent aphthous stomatitis disappeared in 89% of the patients after 1 year of gluten-free diet. Multi logistic analysis selected the following variables as the most meaningful in coeliac disease patients: dental enamel defects (OR = 2.652 CI = 1.427-4.926) and soft tissue lesions (OR = 41.667, CI = 18.868-90.909). Artificial Neural Networks methodology showed that oral soft tissue lesions have sensitivity = 42%, specificity = 98% and test accuracy = 83% in coeliac disease diagnosis. CONCLUSIONS: The overall prevalence of oral soft tissue lesions was higher in coeliac disease patients (42%) than in controls. However, the positive-predictive value of these lesions for coeliac disease diagnosis was low. PMID- 17919277 TI - Meta-analysis: gastrointestinal bleeding due to interaction between selective serotonin uptake inhibitors and non-steroidal anti-inflammatory drugs. AB - BACKGROUND: Selective serotonin reuptake inhibitors (SSRIs) have been associated with upper gastrointestinal haemorrhage (UGIH) but the magnitude and characteristics of this reaction and possible interaction with concurrent Non Steroidal Anti-Inflammatory Drug (NSAID) therapy are unknown. AIM: To evaluate systematically the risk of UGIH with SSRIs, including interaction with NSAIDs. METHODS: We searched PubMED, Science Citation Index, and trial registries for data on SSRIs, NSAIDs and UGIH. We evaluated spontaneous case reports from pharmacovigilance databases. RESULTS: Random effects meta-analysis of four observational studies involving 153 000 patients showed an odds ratio of 2.36 (95% CI: 1.44-3.85; P = 0.0006) for SSRI associated UGIH. The odds ratio increased to 6.33 (95% CI: 3.40-11.8; P < 0.00001) with concomitant NSAIDs. In patients aged above 50 years with no UGIH risk factors, the Number-Needed-to-Harm per year is 411 for SSRIs alone, and 106 with concomitant NSAIDs. Analysis of 101 spontaneous reports showed that UGIH occurred after a median of 25 weeks with SSRIs. Around 67% of these patients were on NSAIDs. CONCLUSIONS: Selective serotonin reuptake inhibitor use, alone and in combination with NSAIDs, substantially increases the risk of UGIH. Clinicians should consider this when managing patients at risk of, or presenting with UGIH. PMID- 17919279 TI - Function of heterologous and truncated RNase P proteins in Bacillus subtilis. AB - Bacterial RNase P is composed of an RNA subunit and a single protein (encoded by the rnpB and rnpA genes respectively). The Bacillus subtilis rnpA knockdown strain d7 was used to screen for functional conservation among bacterial RNase P proteins from a representative spectrum of bacterial subphyla. We demonstrate conserved function of bacterial RNase P (RnpA) proteins despite low sequence conservation. Even rnpA genes from psychrophilic and thermophilic bacteria rescued growth of B. subtilis d7 bacteria; likewise, terminal extensions and insertions between beta strands 2 and 3, in the so-called metal binding loop, were compatible with RnpA function in B. subtilis. A deletion analysis of B. subtilis RnpA defined the structural elements essential for bacterial RNase P function in vivo. We further extended our complementation analysis in B. subtilis strain d7 to the four individual RNase P protein subunits from three different Archaea, as well as to human Rpp21 and Rpp29 as representatives of eukaryal RNase P. None of these non-bacterial RNase P proteins showed any evidence of being able to replace the B. subtilis RNase P protein in vivo, supporting the notion that archaeal/eukaryal RNase P proteins are evolutionary unrelated to the bacterial RnpA protein. PMID- 17919278 TI - Nitrosative stress treatment of E. coli targets distinct set of thiol-containing proteins. AB - Reactive nitrogen species (RNS) function as powerful antimicrobials in host defence, but so far little is known about their bacterial targets. In this study, we set out to identify Escherichia coli proteins with RNS-sensitive cysteines. We found that only a very select set of proteins contain cysteines that undergo reversible thiol modifications upon nitric oxide (NO) treatment in vivo. Of the 10 proteins that we identified, six (AtpA, AceF, FabB, GapA, IlvC, TufA) have been shown to harbour functionally important thiol groups and are encoded by genes that are considered essential under our growth conditions. Media supplementation studies suggested that inactivation of AceF and IlvC is, in part, responsible for the observed NO-induced growth inhibition, indicating that RNS mediated modifications play important physiological roles. Interestingly, the majority of RNS-sensitive E. coli proteins differ from E. coli proteins that harbour H2O2-sensitive thiol groups, implying that reactive oxygen and nitrogen species affect distinct physiological processes in bacteria. We confirmed this specificity by analysing the activity of one of our target proteins, the small subunit of glutamate synthase. In vivo and in vitro activity studies confirmed that glutamate synthase rapidly inactivates upon NO treatment but is resistant towards other oxidative stressors. PMID- 17919280 TI - RutR is the uracil/thymine-sensing master regulator of a set of genes for synthesis and degradation of pyrimidines. AB - Using the genomic SELEX, a total of six Escherichia coli DNA fragments have been identified, which formed complexes with transcription factor RutR. The RutR regulon was found to include a large number of genes encoding components for not only degradation of pyrimidines but also transport of glutamate, synthesis of glutamine, synthesis of pyrimidine nucleotides and arginine, and degradation of purines. DNase I footprinting indicated that RutR recognizes a palindromic sequence of TTGACCAnnTGGTCAA. The RutR box in P1 promoter of carAB encoding carbamoyl phosphate synthetase, a key enzyme of pyrimidine synthesis, overlaps with the PepA (CarP) repressor binding site, implying competition between RutR and PepA. Adding either uracil or thymine abolished RutR binding in vitro to the carAB P1 promoter. Accordingly, in the rutR-deletion mutant or in the presence of uracil, the activation in vivo of carAB P1 promoter was markedly reduced. Northern blot analysis of the RutR target genes indicated that RutR represses the Gad system genes involved in glutamate-dependent acid resistance and allantoin degradation. Altogether we propose that RutR is the pyrimidine sensor and the master regulator for a large set of the genes involved in the synthesis and degradation of pyrimidines. PMID- 17919281 TI - Genetic basis for retention of a critical virulence plasmid of Borrelia burgdorferi. AB - The genome of Borrelia burgdorferi is composed of one linear chromosome and approximately 20 linear and circular plasmids. Although some plasmids are required by B. burgdorferi in vivo, most plasmids are dispensable for growth in vitro. However, circular plasmid (cp) 26 is present in all natural isolates and has never been lost during in vitro growth. This plasmid carries ospC, which is critical for mammalian infection. We previously showed that cp26 encodes essential functions, including the telomere resolvase, ResT, and hence cannot be displaced. Here we identify two additional essential genes on cp26, bbb26 and bbb27, through a systematic attempt to inactivate each open reading frame (ORF). Furthermore, an incompatible plasmid carrying resT, bbb26 and bbb27 could displace cp26. Computational and experimental analyses suggested that both BBB26 and BBB27 are membrane-associated, periplasmic proteins. These data indicate that bbb26 and bbb27 encode essential but possibly redundant functions and that one or the other of these cp26 genes, in addition to resT, is required for bacterial viability. We conclude that the genetic linkage of critical physiological and virulence functions on cp26 is pertinent to its stable maintenance throughout the evolution of B. burgdorferi. PMID- 17919282 TI - The Hsp70 chaperone machines of Escherichia coli: a paradigm for the repartition of chaperone functions. AB - Molecular chaperones are highly conserved in all free-living organisms. There are many types of chaperones, and most are conveniently grouped into families. Genome sequencing has revealed that many organisms contain multiple members of both the DnaK (Hsp70) family and their partner J-domain protein (JDP) cochaperone, belonging to the DnaJ (Hsp40) family. Escherichia coli K-12 encodes three Hsp70 genes and six JDP genes. The coexistence of these chaperones in the same cytosol suggests that certain chaperone-cochaperone interactions are permitted, and that chaperone tasks and their regulation have become specialized over the course of evolution. Extensive genetic and biochemical analyses have greatly expanded knowledge of chaperone tasking in this organism. In particular, recent advances in structure determination have led to significant insights of the underlying complexities and functional elegance of the Hsp70 chaperone machine. PMID- 17919283 TI - Housekeeping sortase facilitates the cell wall anchoring of pilus polymers in Corynebacterium diphtheriae. AB - Many surface proteins in Gram-positive bacteria are covalently linked to the cell wall through a transpeptidation reaction catalysed by the enzyme sortase. Corynebacterium diphtheriae encodes six sortases, five of which are devoted to the assembly of three distinct types of pilus fibres--SrtA for the SpaA-type pilus, SrtB/SrtC for the SpaD-type pilus, and SrtD/SrtE for the SpaH-type pilus. We demonstrate here the function of SrtF, the so-called housekeeping sortase, in the cell wall anchoring of pili. We show that a multiple deletion mutant strain expressing only SrtA secretes a large portion of SpaA polymers into the culture medium, with concomitant decrease in the cell wall-linked pili. The same phenotype is observed with the mutant that is missing SrtF alone. By contrast, a strain that expresses only SrtF displays surface-linked pilins but no polymers. Therefore, SrtF can catalyse the cell wall anchoring of pilin monomers as well as pili, but it does not polymerize pilins. We show that SrtA and SrtF together generate wild-type levels of the SpaA-type pilus on the bacterial surface. Furthermore, by regulating the expression of SpaA in the cell, we demonstrate that the SrtF function becomes critical when the SpaA level is sufficiently high. Together, these findings provide key evidence for a two-stage model of pilus assembly: pilins are first polymerized by a pilus-specific sortase, and the resulting fibre is then attached to the cell wall by either the cognate sortase or the housekeeping sortase. PMID- 17919284 TI - GolS controls the response to gold by the hierarchical induction of Salmonella specific genes that include a CBA efflux-coding operon. AB - Salmonella employs a specific set of proteins that allows it to detect the presence of gold salts in the environment and to mount the appropriate resistance response. This includes a P-type ATPase, GolT, and a small cytoplasmic metal binding protein, GolB. Their expression is controlled by a MerR-like sensor, GolS, which is highly selective for Au ions. Here, we identify a new GolS controlled operon named gesABC which codes for a CBA efflux system, and establish its role in Au resistance. GesABC can also mediate drug resistance when induced by Au in a GolS-dependent manner, in a strain deleted in the main drug transporter acrAB. The GolS-controlled transcription of gesABC differs from the other GolS-regulated loci. It is activated by gold, but not induced by copper, even in a strain deleted of the main Cu transporter gene copA, which triggers a substantial GolS-dependent induction of golTS and golB. We demonstrate that the Au-dependent induction of gesABC transcription requires higher GolS levels than for the other members of the gol regulon. This correlates with a divergent GolS operator in the gesABC promoter. We propose that the hierarchical induction within the gol regulon allows Salmonella to cope with Au-contaminated environments. PMID- 17919285 TI - Glutathione and Gts1p drive beneficial variability in the cadmium resistances of individual yeast cells. AB - Phenotypic heterogeneity among individual cells within isogenic populations is widely documented, but its consequences are not well understood. Here, cell-to cell variation in the stress resistance of Saccharomyces cerevisiae, particularly to cadmium, was revealed to depend on the antioxidant glutathione. Heterogeneity was decreased strikingly in gsh1 mutants. Furthermore, cells sorted according to differing reduced-glutathione (GSH) contents exhibited differing stress resistances. The vacuolar GSH-conjugate pathway of detoxification was implicated in heterogeneous Cd resistance. Metabolic oscillations (ultradian rhythms) in yeast are known to modulate single-cell redox and GSH status. Gts1p stabilizes these oscillations and was found to be required for heterogeneous Cd and hydrogen peroxide resistance, through the same pathway as Gsh1p. Expression of GTS1 from a constitutive tet-regulated promoter suppressed oscillations and heterogeneity in GSH content, and resulted in decreased variation in stress resistance. This enabled manipulation of the degree of gene expression noise in cultures. It was shown that cells expressing Gts1p heterogeneously had a competitive advantage over more-homogeneous cell populations (with the same mean Gts1p expression), under continuous and fluctuating stress conditions. The results establish a novel molecular mechanism for single-cell heterogeneity, and demonstrate experimentally fitness advantages that depend on deterministic variation in gene expression within cell populations. PMID- 17919286 TI - A partner for the resuscitation-promoting factors of Mycobacterium tuberculosis. AB - Many cases of active tuberculosis are thought to result from the reactivation of dormant Mycobacterium tuberculosis from a prior infection, yet remarkably little is known about the mechanism by which these non-sporulating bacteria reactivate. A family of extracellular bacterial proteins, known as resuscitation-promoting factors (Rpfs), has previously been shown to stimulate growth of dormant mycobacteria. While Rpf proteins are clearly peptidoglycan glycosidases, the mechanism and role of Rpf in mediating reactivation remains unclear. Here we use a yeast two-hybrid screen to identify potential binding partners of RpfB and report the interaction between RpfB and a putative mycobacterial endopeptidase, which we named Rpf-interacting protein A (RipA). This interaction was confirmed by in vitro and in vivo co-precipitation assays. The interacting domains map to the C-termini of both proteins, near predicted enzymatic domains. We show that RipA is a secreted, cell-associated protein, found in the same cellular compartment as RpfB. Both RipA and RpfB localize to the septa of actively growing bacteria by fluorescence microscopy. Finally, we demonstrate that RipA is capable of digesting cell wall material and is indeed a peptidoglycan hydrolase. The interaction between these two peptidoglycan hydrolases at the septum suggests a role for the complex in cell division, possibly during reactivation. PMID- 17919287 TI - Regulation of fatty acid metabolism in bacteria. AB - In Escherichia coli, the main player in transcription regulation of fatty acid metabolism is the FadR protein, which is involved in negative regulation of fatty acid degradation and in positive and negative regulation of the cellular processes related to it, as well as in positive regulation of the biosynthesis of unsaturated fatty acids in a concerted manner with negative regulation of FabR. On the other hand, Bacillus subtilis possesses two global transcriptional regulators, FadR (YsiA) and FapR. B. subtilis FadR represses fatty acid degradation, whereas FapR represses almost all the processes in the biosynthesis of saturated fatty acids and phospholipids. Furthermore, Streptococcus pneumoniae FabT represses the genes of fatty acid biosynthesis that are clustered in its genome. Long-chain acyl-CoAs appear to be metabolic signals for fatty acid degradation by bacteria in general, and antagonize the FadR protein from either E. coli or B. subtilis. However, malonyl-CoA is a metabolic signal for fatty acid and phospholipid biosynthesis by Gram-positive low-GC bacteria, and it antagonizes FapR. These would be the primary aspects for understanding the elaborate and complex regulation of fatty acid metabolism in bacteria to maintain membrane lipid homeostasis. PMID- 17919289 TI - The initiation and pattern of spread of histone H4 acetylation parallel the order of transcriptional activation of genes in the aflatoxin cluster. AB - The 27 genes involved in aflatoxin biosynthesis are clustered within a 70 kb region in the Aspergillus parasiticus genome. Using chromatin immunoprecipitation, we demonstrated a positive correlation between the initiation and spread of histone H4 acetylation in aflatoxin promoters and the onset of accumulation of aflatoxin proteins and aflatoxin. Histone H4 acetylation in the pksA (encodes an 'early' biosynthetic pathway enzyme) promoter peaked at 30 h, prior to the increased acetylation in the omtA and ordA (encode 'late' enzymes) promoters detected at 40 h. The specific order in which pksA, ver-1 (encodes a 'middle' enzyme) and omtA transcripts accumulated in cells paralleled the pattern of spread of histone H4 acetylation. Binding of AflR, a positive regulator of aflatoxin biosynthesis, to the ordA promoter showed a positive correlation with the spread of histone H4 acetylation. The data suggest that the order of genes within the aflatoxin cluster determines the timing and order of transcriptional activation, and that the site of initiation and spread of histone H4 acetylation mediate this process. Our data indicate that the aflatoxin and adjacent sugar utilization clusters are part of a larger 'regulatory unit'. PMID- 17919288 TI - Interaction of Bacteroides fragilis pLV22a relaxase and transfer DNA with Escherichia coli RP4-TraG coupling protein. AB - Many Bacteroides transfer factors are mobilizable in Escherichia coli when coresident with the IncP conjugative plasmid RP4, but not F. To begin characterization and potential interaction between Bacteroides mobilizable transfer factors and the RP4 mating channel, both mutants and deletions of the DNA processing (dtr), mating pair formation (mpf) and traG coupling genes of RP4 were tested for mobilization of Bacteroides plasmid pLV22a. All 10 mpf but none of the four dtr genes were required for mobilization of pLV22a. The RP4 TraG coupling protein (CP) was also required for mobilization of pLV22a, but could be substituted by a C-terminal deletion mutant of the F TraD CP. Potential interactions of the TraG CP with relaxase protein(s) and transfer DNA of both RP4 and pLV22a were assessed. Overlay assays identified productive interactions between TraG and the relaxase proteins of both MbpB and TraI from pLV22a and RP4 respectively. The Agrobacterium Transfer-ImmunoPrecipitation (TrIP) assay also identified an interaction between TraG and both RP4 and pLV22a transfer DNA. Thus, mobilization of the Bacteroides pLV22a in E. coli utilizes both RP4 Mpf and CP functions including an interaction between the relaxosome and the RP4 CP similar to that of cognate RP4 plasmid. PMID- 17919290 TI - The AraC/XylS regulator TxtR modulates thaxtomin biosynthesis and virulence in Streptomyces scabies. AB - Streptomyces scabies is the best studied of those streptomycetes that cause an economically important disease known as potato scab. The phytotoxin thaxtomin is made exclusively by these pathogens and is required for virulence. Here we describe regulation of thaxtomin biosynthesis by TxtR, a member of the AraC/XylS family of transcriptional regulators. The txtR gene is imbedded in the thaxtomin biosynthetic pathway and is located on a conserved pathogenicity island in S. scabies, S. turgidiscabies and S. acidiscabies. Thaxtomin biosynthesis was abolished and virulence was almost eliminated in the txtR deletion mutant of S. scabies 87.22. Accumulation of thaxtomin biosynthetic gene (txtA, txtB, txtC, nos) transcripts was reduced compared with the wild-type S. scabies 87.22. NOS dependent nitric oxide production by S. scabies was also reduced in the mutant. The TxtR protein bound cellobiose, an inducer of thaxtomin production, and transcription of txtR and thaxtomin biosynthetic genes was upregulated in response to cellobiose. TxtR is the first example of an AraC/XylS family protein regulated by cellobiose. Together, these data suggest that cellobiose, the smallest oligomer of cellulose, may signal the availability of expanding plant tissue, which is the site of action of thaxtomin. PMID- 17919291 TI - Assessment of bronchial wall thickness and lumen diameter in human adults using multi-detector computed tomography: comparison with theoretical models. AB - A thickened bronchial wall is the morphological substratum of most diseases of the airway. Theoretical and clinical models of bronchial morphometry have so far focused on bronchial lumen diameter, and bronchial length and angles, mainly assessed from bronchial casts. However, these models do not provide information on bronchial wall thickness. This paper reports in vivo values of cross-sectional wall area, lumen area, wall thickness and lumen diameter in ten healthy subjects as assessed by multi-detector computed tomography. A validated dedicated software package was used to measure these morphometric parameters up to the 14th bronchial generation, with respect to Weibel's model of bronchial morphometry, and up to the 12th according to Boyden's classification. Measured lumen diameters and homothety ratios were compared with theoretical values obtained from previously published studies, and no difference was found when considering dichotomic division of the bronchial tree. Mean wall area, lumen area, wall thickness and lumen diameter were then provided according to bronchial generation order, and mean homothety ratios were computed for wall area, lumen area and wall thickness as well as equations giving the mean value of each parameter for a given bronchial generation with respect to its value in generation 0 (trachea). Multi-detector computed tomography measurements of bronchial morphometric parameters may help to improve our knowledge of bronchial anatomy in vivo, our understanding of the pathophysiology of bronchial diseases and the evaluation of pharmacological effects on the bronchial wall. PMID- 17919292 TI - Different structural stability and toxicity of PrP(ARR) and PrP(ARQ) sheep prion protein variants. AB - The polymorphisms at amino acid residues 136, 154, and 171 in ovine prion protein (PrP) have been associated with different susceptibility to scrapie: animals expressing PrP(ARQ) [PrP(Ala136/Arg154/Gln171)] show vulnerability, whereas those that express PrP(ARR) [PrP(Ala136/Arg154/Arg171)] are resistant to scrapie. The aim of this study was to evaluate the in vitro toxic effects of PrP(ARR) and PrP(ARQ) variants in relation with their structural characteristics. We show that both peptides cause cell death inducing apoptosis but, unexpectedly, the scrapie resistant PrP(ARR) form was more toxic than the scrapie susceptible PrP(ARQ) variant. Moreover, the alpha-helical conformation of PrP(ARR) was less stable than that of PrP(ARQ) and the structural determinants responsible of these different conformational stabilities were characterized by spectroscopic analysis. We observed that PrP toxicity was inversely related to protein structural stability, being the unfolded conformation more toxic than the native one. However, the PrP(ARQ) variant displays a higher propensity to form large aggregates than PrP(ARR). Interestingly, in the presence of small amounts of PrP(ARR), PrP(ARQ) aggregability was reduced to levels similar to that of PrP(ARR). Thus, in contrast to PrP(ARR) toxicity, scrapie transmissibility seems to reside in the more stable conformation of PrP(ARQ) that allows the formation of large amyloid fibrils. PMID- 17919293 TI - Proteomic analysis of the effects and interactions of sleep deprivation and aging in mouse cerebral cortex. AB - The cellular and molecular processes that underlie the drives and functions of sleep have been the topic of many studies in the last few decades. Discovery based techniques, such as cDNA microarrays, have increasingly been utilized in conjunction with sleep deprivation paradigms to examine the molecular mechanisms and functions of sleep. These studies have helped to validate and expand existing hypotheses, such as those on the roles of sleep in synaptic plasticity and in energy metabolism. The mechanisms underlying the highly prevalent changes in sleep architecture with age are not known, but likely reflect fundamental changes in the molecular basis of circadian timing and sleep homeostatic processes. We decided to explore the effects and interactions of sleep deprivation and aging utilizing the proteomic technique of difference in gel electrophoresis (DIGE). DIGE, which utilizes cyanine dye labeling of samples, allows for the comparison of multiple experimental groups within and across gels. In this study, we compared cerebral cortex tissue from young (2.5 months) and old (24 months) mice that had been sleep deprived for 6 h to tissue from undisturbed young and old control animals. Following DIGE, automatic image matching and spot identification, and statistical analysis, 43 unique proteins were identified. The proteins were grouped into seven functional classes based on published characteristics: cell signaling, cytoskeletal, energy metabolism, exocytosis, heat shock proteins, mRNA processing/trafficking, and serum proteins. The identity and characteristics of these proteins relevant to sleep and aging are discussed. PMID- 17919294 TI - Preliminary results of a limited thoracotomy: new approach to treat atrial fibrillation. AB - OBJECTIVE: To determine efficacy of a new procedure combining epicardial bipolar radiofrequency (RF) pulmonary vein (PV) antrum isolation and ganglionated plexus (GP) ablation for treatment of atrial fibrillation (AF). BACKGROUND: PV antrum electrical isolation and GP ablation have each been associated with elimination of AF. Both of these can be performed epicardially in a single combined surgical procedure, which may have advantages over endocardial ablation. METHODS AND RESULTS: Twenty-one subjects entered a prospective evaluation of limited thoracotomy epicardial bipolar PV antrum isolation, verified by PV recordings, with GP ablation, guided by GP mapping. Procedural success was defined as freedom from AF and antiarrhythmic agents during 1 year of follow-up, including evaluation by prolonged continuous monitoring capable of detecting asymptomatic arrhythmias. All subjects had recordable PV potentials and GP activity prior to ablation. Circumferential epicardial bipolar RF eliminated PV potentials in 18 of 20 right and 14 of 20 left PV antra. This concurrently eliminated 79% of GP activity (125 of 159 active sites); nearly all remaining GP activity could then be eliminated using epicardial bipolar RF forceps. Fifteen of 20 (75%) subjects overall, and 14 of 16 (87.5%) subjects with paroxysmal or persistent AF had a successful procedure. CONCLUSION: Limited thoracotomy epicardial bipolar RF antrum isolation, verified by PV recordings, with GP ablation, guided by GP mapping, is effective treatment for AF and should be considered in patients with paroxysmal or persistent AF. PMID- 17919295 TI - Failure to pace: pacemaker malfunction? PMID- 17919297 TI - Host response to Helicobacter pylori infection before initiation of the adaptive immune response. AB - Helicobacter pylori persistently colonizes the human stomach. In this study, immune responses to H. pylori that occur in the early stages of infection were investigated. Within the first 2 days after orogastric infection of mice with H. pylori, there was a transient infiltration of macrophages and neutrophils into the glandular stomach. By day 10 postinfection, the numbers of macrophages and neutrophils decreased to baseline levels. By 3 weeks postinfection, an adaptive immune response was detected, marked by gastric infiltration of T lymphocytes, macrophages, and neutrophils, as well as increased numbers of H. pylori-specific T cells, macrophages, and dendritic cells in paragastric lymph nodes. Neutrophil attracting and macrophage-attracting chemokines were expressed at higher levels in the stomachs of H. pylori-infected mice than in the stomachs of uninfected mice. Increased expression of TNFalpha and IFNgamma (Th1-type inflammatory cytokines) and IL-17 (a Th17-type cytokine) was detected in the stomachs of H. pylori-infected mice, but increased expression of IL-4 (a Th2-type cytokine) was not detected. These data indicate that a transient gastric inflammatory response to H. pylori occurs within the first few days after infection, before the priming of T cells and initiation of an adaptive immune response. It is speculated that inappropriate waning of the innate immune response during early stages of infection may be a factor that contributes to H. pylori persistence. PMID- 17919298 TI - Differences in faecal bacterial communities in coeliac and healthy children as detected by PCR and denaturing gradient gel electrophoresis. AB - Coeliac disease (CD) is a chronic inflammatory disorder of the small intestinal mucosa. Scientific evidence supports a role of the gut microbiota in chronic inflammatory disorders; yet information is not specifically available for CD. In this study, a comparative denaturing gradient gel electrophoresis analysis of faecal samples from coeliac children and age-matched controls was carried out. The diversity of the faecal microbiota was significantly higher in coeliac children than in healthy controls. The presence of the species Lactobacillus curvatus, Leuconostoc mesenteroides and Leuconostoc carnosum was characteristic of coeliac patients, while that of the Lactobacillus casei group was characteristic of healthy controls. The Bifidobacterium population showed a significantly higher species diversity in healthy children than in coeliacs. In healthy children, this population was characterized by the presence of Bifidobacterium adolescentis. Overall, the results highlighted the need for further characterization of the microbiota in coeliac patients, and suggested a potential role of probiotics and/or prebiotics in restoring their gut microbial balance. PMID- 17919296 TI - Heritability of specific language impairment depends on diagnostic criteria. AB - Heritability estimates for specific language impairment (SLI) have been inconsistent. Four twin studies reported heritability of 0.5 or more, but a recent report from the Twins Early Development Study found negligible genetic influence in 4-year-olds. We considered whether the method of ascertainment influenced results and found substantially higher heritability if SLI was defined in terms of referral to speech and language pathology services than if defined by language test scores. Further analysis showed that presence of speech difficulties played a major role in determining whether a child had contact with services. Childhood language disorders that are identified by population screening are likely to have a different phenotype and different etiology from clinically referred cases. Genetic studies are more likely to find high heritability if they focus on cases who have speech difficulties and who have been referred for intervention. PMID- 17919299 TI - Recombinant BCG coexpressing Ag85B, ESAT-6 and mouse-IFN-gamma confers effective protection against Mycobacterium tuberculosis in C57BL/6 mice. AB - In this study, the protective efficacy of a novel recombinant bacille Calmette Geurin (BCG) strain (rBCG-AEI) expressing fusion protein the antigen 85B (Ag85B)- the 6-kDa early secreted antigen target (ESAT-6)-IFN-gamma against Mycobacterium tuberculosis H37Rv in mice was evaluated. The immunogenicity study showed that rBCG-AEI could induce higher specific antibody titers and significantly increase cellular immune response than BCG, or rBCG-A strain (expressing Ag85B), or rBCG AE strain (expressing fusion protein Ag85B-ESAT-6). The protective experiment demonstrated that rBCG-AEI could confer similar or even better protective efficacy against M. tuberculosis infection compared with others in organ bacterial loads, lung histopathology and net weight gain or loss. The results suggested that rBCG-AEI is a potential candidate for further study. PMID- 17919300 TI - Siderophore-mediated cooperation and virulence in Pseudomonas aeruginosa. AB - Why should organisms cooperate with each other? Helping close relatives that are likely to share the same genes (kin selection) is one important explanation that is likely to apply across taxa. The production of metabolically costly extracellular iron-scavenging molecules (siderophores) by microorganisms is a cooperative behaviour because it benefits nearby conspecifics. We review experiments focusing on the production of the primary siderophore (pyoverdin) of the opportunistic bacterial pathogen, Pseudomonas aeruginosa, which test kin selection theories that seek to explain the evolution of cooperation. First, cooperation is indeed favoured when individuals interact with their close relatives and when there is competition between groups of cooperators and noncooperators, such that the benefit of cooperation can be realized. Second, the relative success of cheats and cooperators is a function of their frequencies within populations. Third, elevated mutation rates can confer a selective disadvantage under conditions when cooperation is beneficial, because high mutation rates reduce how closely bacteria are related to each other. Fourth, cooperative pyoverdin production is also shown to be favoured by kin selection in vivo (caterpillars), and results in more virulent infections. Finally, we briefly outline ongoing and future work using this experimental system. PMID- 17919301 TI - A prospective study of the modified Atkins diet for intractable epilepsy in adults. AB - PURPOSE: The ketogenic diet is not typically offered to adults with epilepsy due to the significant lifestyle alterations needed for its use. The modified Atkins diet has been recently demonstrated to be therapeutic for children without the need for an admission, fasting period, weighing of foods, or fluid, calorie, and protein restriction. METHODS: A prospective, open-label study was performed of adults over 18 years of age, having at least weekly seizures and prior use of at least two anticonvulsants. Carbohydrates were initially restricted to 15 g/day, fats were encouraged, and fluids, protein, and calories were allowed ad lib. RESULTS: Thirty patients, with age ranging from 18 to 53 years, were enrolled. Using an intent-to-treat analysis, 47% had a >50% seizure reduction after 1 and 3 months on the diet; 33% after 6 months. In those with seizure reduction, the median time to improvement was 2 weeks (range: 1-8 weeks). The mean weight loss was 6.8 kg, p < 0.001. Body-mass index (BMI) decrease correlated with efficacy at 3 months, p = 0.03. Ten subjects (30%) discontinued the diet prior to 3 months. Side effects included increased cholesterol (mean 187 to 201 mg/dL), blood urea nitrogen (BUN; 13 to 16 mg/dL), and urine calcium to creatinine ratio (0.14 to 0.19). CONCLUSIONS: A modified Atkins diet appears to demonstrate preliminary efficacy for adults with intractable epilepsy, especially in those who lost weight. Considering the rapid response in those who improved, but somewhat high discontinuation rate, a 2-month trial period may be adequate to assess for efficacy. PMID- 17919303 TI - Contact allergy and medicinal herbs. AB - Herbal treatments are becoming increasingly popular, and are often used for internal as well as dermatological conditions, both externally as well as orally. The prevalence of contact sensitization against several plants especially of the Compositae family is quite high in Europe. Sensitization seems to occur relatively frequent with a few species such as arnica, elecampane and tea tree (oil), and occurs rarely with the majority. Testing for plant allergy is problematic because of the limited number of commercially available standardized patch test substances and the danger of active sensitization when testing with plants, parts thereof, or individual extracts. Knowledge about the allergic potential of plants is limited. Although plants are regarded as critical allergens by dermatologists, the number of reported cases of contact dermatitis is relatively small. Many widely used substances are not licensed as drugs or cosmetics. While the positive effects are frequently questionable or limited, the side effects are often more evident. Adverse effects of herbal medicines are an important albeit neglected subject in dermatology, which deserves further systematic investigation. PMID- 17919302 TI - The NR1 N-methyl-D-aspartate subunit and brain-derived neurotrophic factor in temporal lobe epilepsy hippocampus: a comparison of patients with and without coexisting psychiatric symptoms. AB - PURPOSE: The glutamate N-methyl-D-aspartate (NMDA) receptor and the neurotrophin brain-derived neurotrophic factor have been implicated in the pathophysiology of schizophrenia and depression. Since these psychiatric disorders are common in temporal lobe epilepsy (TLE), a comparison of TLE patients with and without coexisting psychiatric symptoms may be useful to unravel pathophysiologic mechanisms for psychosis or depression. METHODS: We used immunoautoradiography to assess the NR1 NMDA receptor subunit and brain-derived neurotrophic factor in resected TLE hippocampus. RESULTS: No changes relative to comparison controls were found for TLE patients with schizophrenia-like psychosis or depression. Increased NR1 was found in the dentate molecular layer in the dysphoria group and unmedicated depressed patients. CONCLUSIONS: An increase in NR1 protein in the dentate molecular layer suggests an upregulation of NMDA receptors in granule cells in TLE patients with dysphoria and depression. This finding is compatible with the theory that increased NMDA receptor function is involved in the pathogenesis of depression and that antidepressants may act by opposing this mechanism. PMID- 17919304 TI - Particularities in managing melanoma patients with tattoos: case report and review of the literature. AB - Assessing pigmented lesions within or covered by tattoos is difficult, both at a clinical and dermatoscopic level. Patients with tattoos may provide a challenge in the early recognition and follow-up of melanoma. PMID- 17919305 TI - Headache associated with sexual activity: prognosis and treatment options. AB - The aim of this study was to provide data on the prognosis and treatment options of headache associated with sexual activity (HSA). Sixty patients diagnosed with HSA between 1996 and 2004 were followed up between 2003 and 2006 at least 12 months after the first interview. The further course of the disease and their contentedness with therapy were requested. On average, the second interview was performed 35.9 months after the first examination. Of the 45 patients who had suffered from single attacks or bouts prior to baseline examination, 37 had no further attacks. Seven patients suffered from at least one further bout with an average duration of 2.1 months. One patient developed a chronic course of the disease after an episodic start. Of the 15 patients with chronic disease at the first examination, seven were in remission and five had ongoing attacks at follow up. Ten patients received indomethacin for preemptive therapy, with good results in nine patients. Eighteen patients received beta-blockers for prophylaxis, with good results in 15 patients. Episodic HSA occurs in approximately three-quarters and chronic HSA in approximately one-quarter of patients. Even in chronic HAS, the prognosis is favourable, with remission rates of 69% during an observation period of 3 years. For patients with longer-lasting bouts or with chronic HSA, prophylactic treatment with beta-blockers or preemptive therapy with indomethacin are often successful. PMID- 17919306 TI - Seasonal variation and atypical presentation of idiopathic intracranial hypertension in pre-pubertal children. AB - Idiopathic intracranial hypertension is an enigmatic disorder of elevated cerebrospinal fluid pressure. In adulthood, patients are typically obese women of childbearing age; however, in young children the clinical picture is strikingly different, indicating age-related differences in the aetiology of idiopathic intracranial hypertension. To investigate this phenomenon, we analysed the clinical details of 15 pre-pubertal children with the diagnosis of idiopathic intracranial hypertension. Evaluating the date of initial presentation, we discovered a distinct seasonal variation. Ten patients presented between November and March, thus coinciding with the typical season of paediatric viral and bacterial infections in Germany. Therefore, we suggest an association between intracranial hypertension and possibly concurrent infections in these children. Moreover, eight children presented only with ophthalmologic findings without any other obvious symptoms, raising questions regarding the incidence of undetected cases, particularly in this age group. PMID- 17919307 TI - Investigation of neuromuscular transmission in some rare types of migraine. AB - The aim of this study was to delineate any dysfunction of neuromuscular transmission (NMT) by single-fibre electromyography (SFEMG) in some rare types of migraine. Recent studies have shown subclinical dysfunction of NMT in migraine with aura and cluster headache by using SFEMG, whereas another recent study has shown NMT to be normal in familial hemiplegic migraine (FHM) with CACNA1A mutations. Thirty patients with rare primary headache syndromes [18 with sporadic hemiplegic migraine (SHM), six with FHM and six with basilar-type migraine (BM)] and 15 healthy control subjects without any headache complaints underwent nerve conduction studies, EMG and SFEMG during voluntary contraction of the extensor digitorum communis muscle. Ten to 20 different potential pairs were recorded and individual jitter values calculated. The results obtained from patient groups were compared with those from the normal subjects. Of 600 individual jitter values of the patients, 27 (4.5%) were abnormally high, whereas only 3/205 (1.5%) jitter values from normal subjects were abnormal. Abnormal NMT was found in 4/30 (13.3%) patients (three SHM and one BM), but in none of the control subjects. Only in SHM patients was the number of individual abnormal jitter values slightly but significantly different from normal controls. The present study demonstrates that subclinical NMT abnormality is slightly present in only SHM and BM patients, but not in FHM patients. PMID- 17919308 TI - Residential exposure to mould and dampness is associated with adverse respiratory health. AB - BACKGROUND: Indoor exposure to mould and dampness is frequently associated with asthma symptoms with and without lung function changes. However, the mechanisms contributing to this threat to respiratory health are only partly understood. OBJECTIVE: To investigate the contribution of recent exposure to mould and dampness in the living room or bedroom to respiratory health in a general practice-based cohort of 526 asthmatic children. METHODS: Parents were questioned about home characteristics, including moulds and dampness. The level of asthma control was evaluated in their participating children by means of asthma symptoms, peak expiratory flow (PEF) variability, severity of airway hyperresponsiveness (AHR), and medication usage. RESULTS: Children exposed to indoor moulds and dampness more often had severe AHR compared with non-exposed (42% vs. 16%; P< or =0.001). They also showed an increased PEF variability (11.3% vs. 8.4%; P=0.03) and, however, not significant, more frequent asthma symptoms. The use of controller medication was not significantly different between exposed and non-exposed children. After adjustment for gender, age, smoking, exposure to parental smoking, parental education, pet ownership, presence of inhalant allergy, use of controller medication, health care center, and season of study assessment, the odds ratio for severe AHR in exposed children was 3.95 [95% confidence interval (CI): 1.82-8.57]. CONCLUSION: We found a consistent association between reported moulds and dampness in the living room or the child's bedroom and an increased risk for severe AHR in a general practice-based cohort of asthmatic children, even after adjustment for gender, presence of inhalant allergy, and use of controller medication. PMID- 17919309 TI - Non-neuronopathic Gaucher disease due to saposin C deficiency. AB - Gaucher disease is generally caused by a deficiency of the lysosomal enzyme glucocerebrosidase. The degradation of glycosphingolipids requires also the participation of sphingolipid activator proteins. The prosaposin PSAP gene codes for a single protein which undergoes post-translational cleavage to yield four proteins named saposins A, B, C and D. Saposin (SAP-) C is required for glucosylceramide degradation, and its deficiency results in a variant form of Gaucher disease. In this report, we present clinical, biochemical, and molecular findings in a 36-year-old man and his 30-year-old sister with non-neuronopathic Gaucher disease due to SAP-C deficiency. Very high levels of chitotriosidase activity, chemokine CCL18, and increased concentration of glucosylceramide in plasma and normal beta-glucosidase activity in skin fibroblasts were observed in the patients. A molecular genetics study of the PSAP gene enabled the identification of one missense mutation, p.L349P, located in the SAP-C domain and another mutation, p.M1L, located in the initiation codon of the prosaposin precursor protein. The presented findings describe the first cases where the non neuronopathic Gaucher disease has been definitely demonstrated to be a consequence of SAP-C deficiency. Three previously described cases in the literature displayed a Gaucher type 3 phenotype. PMID- 17919310 TI - Phase I trial of personalized peptide vaccination for cytokine-refractory metastatic renal cell carcinoma patients. AB - The aim of this clinical trial was to investigate the toxicity and immunological responses of personalized peptide vaccination for cytokine-refractory metastatic renal cell carcinoma patients. Patients were confirmed to be human leukocyte antigen (HLA)-A24 or HLA-A2 positive and had histologically confirmed renal cell carcinoma. Ten patients were enrolled in the present study. The peptides to be administered were determined based on the presence of peptide-specific cytotoxic T lymphocyte precursors in peripheral blood mononuclear cells (PBMC) and peptide specific IgG in the plasma of cancer patients. Patients received subcutaneous injections of four different peptides (3 mg/peptide) every 2 weeks. Vaccinations were well tolerated without any major adverse events. A minimal increase in peptide-specific interferon-gamma production in postvaccination PBMC was observed, regardless of higher levels of cytotoxic T lymphocyte activity in prevaccination PBMC. In contrast, an increase in peptide-specific IgG levels of postvaccination (sixth) plasma was observed in the majority of patients. After progression, five patients received interleukin-2 therapy and continuous vaccination, with survival of 31, 25, 23, 17, and 15 months, but interleukin-2 did not impede humoral responses boosted by the vaccination. These results encourage further clinical trials of personalized peptide vaccinations. PMID- 17919311 TI - Expression of the Rai (Shc C) adaptor protein in the human enteric nervous system. AB - The adaptor protein Rai (ShcC/N-Shc) is almost exclusively present in the nervous system, although little is documented about its expression in the gut and the enteric nervous system (ENS). As Rai is a physiological substrate of Ret, an important factor for the development of ENS, we have evaluated the expression of Rai in the ENS in various segments of the human gastrointestinal tract. The expression of Rai was assessed by immunohistochemistry in disease-free human gut samples (oesophagus, stomach, small bowel and colon) obtained from subjects undergoing surgical procedures. Rai was not expressed in the epithelia or lymphoid tissue, whereas a moderate level of expression was observed in the endothelial cells of blood vessels and on the outer membrane of smooth muscle cells in both the muscularis mucosae and the muscularis propria. In the ENS, strong positivity was observed only in enteric glial cells, overlapping with GFAP and S100. In conclusion, Rai is expressed in the human gut, especially in the enteric glial cells. We conclude that Rai may provide an additional marker for this cell type. PMID- 17919312 TI - Downregulated hypothalamic 5-HT3 receptor expression and enhanced 5-HT3 receptor antagonist-mediated improvement in fatigue-like behaviour in cholestatic rats. AB - The serotonin neurotransmitter system, including the 5-HT(3) receptor, has been implicated in the genesis of fatigue in patients with liver disease. Therefore, we examined the possible role of 5-HT(3) receptors in cholestasis-associated fatigue. Rats were either bile duct resected (BDR) or sham resected and studied 10 days postsurgery. A significant decrease in hypothalamic 5-HT(3) receptor expression was detected by immunohistochemistry and Western blot in BDR vs sham rats, coupled with increased hypothalamic serotonin turnover identified by an elevated 5-hydroxyindoleacetic acid (5-HIAA) to 5-HT ratio in BDR vs sham rats. To examine fatigue-like behaviour, an activity meter was used. BDR rats exhibited significantly lower locomotor activity than did sham animals. Subcutaneous injection of the 5-HT(3) receptor antagonist tropisetron (0.1 mg kg(-1)) resulted in significantly increased locomotor activity in BDR rats compared to the activity in saline-treated controls, but was without effect in sham rats. However, a 10-fold higher dose of tropisetron significantly increased locomotor activity in both BDR and sham rats compared to saline-injected controls. These findings indicate that cholestasis in the rat is associated with increased hypothalamic serotonin turnover, decreased hypothalamic 5-HT(3) receptor expression, and enhanced sensitivity to locomotor activation induced by 5-HT(3) receptor antagonism, thereby implicating the 5-HT(3) receptor system in cholestasis associated fatigue. PMID- 17919313 TI - Assessment of potential risk factors associated with ischaemic colitis. AB - Ischaemic colitis (IC) has been associated with a number of diverse disorders and risk factors, including irritable bowel syndrome (IBS) and constipation. We sought to assess, through a large-scale population study, the potential risk factors associated with IC. Patients with IC and matched controls without IC were identified using the medical and pharmacy claims data from the HealthCore Managed Care Database from 1st January 2000 to 31st May 2005. A multivariate conditional logistic regression model was developed to identify significant risk factors of IC. Interactions of age, sex, prior IBS diagnosis, and prior constipation diagnosis were further evaluated. We identified 1754 patients with IC and 6970 non-IC controls; 64% were women, and mean ages were 63 and 62 years respectively. The final parsimonious model comprised 19 independent variables associated with increased risk for IC including shock, dysentery, bloating, IBS, colon carcinoma, constipation, cardiovascular disease, dyspepsia, abdominal, aortic, or cardiovascular surgery, 12-month laxative, H(2) receptor blocker and oral contraceptive use. A significant interaction was observed between age and prior IBS on risk for IC. In conclusion, multiple risk factors for IC were identified and we confirmed that patients with IBS or constipation are at greater risk for IC. PMID- 17919314 TI - Sympathetic (electrodermal) activity during repeated maximal rectal distensions in patients with irritable bowel syndrome and constipation. AB - Irritable bowel syndrome (IBS) is associated with visceral hypersensitivity, stress and autonomic dysfunction. Sympathetic activity during repeated events indicates excitatory or inhibitory mechanisms such as sensitization or habituation. We investigated skin conductance (SC) during repetitive rectal distensions at maximal tolerable pressure in patients with IBS and chronic constipation. Twenty-seven IBS patients, 13 constipation patients and 18 controls underwent two sets of isobaric rectal distensions. First, maximal tolerable distension was determined and then it was repeated five times. Skin conductance was measured continuously. Subjective symptom assessment remained steady in all groups. The baseline values of SC were higher in IBS patients than in patients with constipation and significantly lower in constipation patients than in controls. The maximal SC response to repetitive maximal distensions was higher in IBS patients compared with constipation patients. The amplitude of the initial SC response decreased successively with increased number of distensions in patients with IBS and constipation but not in controls. Irritable bowel syndrome and constipation patients habituated to maximal repetitive rectal distensions with decreasing sympathetic activity. Irritable bowel syndrome patients had higher sympathetic reactivity and baseline activity than constipation patients. A lower basal SC in constipation patients compared with controls suggests an inhibition of the sympathetic drive in constipation patients. PMID- 17919315 TI - Lemon juice has protective activity in a rat urolithiasis model. AB - BACKGROUND: The use of herbal medicines (medicinal plants or phytotherapy) has recently gained popularity in Europe and the United States. Nevertheless the exact mechanism of the preventive effects of these products is still far to be clearly established, being its knowledge necessary to successfully apply these therapies to avoid stone formation. METHODS: The effect of oral lemon juice administration on calcium oxalate urolithiasis was studied in male Wistar rats. Rats were rendered nephrolithic by providing drinking water containing 0.75% ethylene glycol [v/v] (EG) and 2% ammonium chloride [w/v] (AC) for 10 days. In addition to EG/AC treatment, three groups of rats were also gavage-administered solutions containing 100%, 75% or 50% lemon juice [v/v] (6 microl solution/g body weight). Positive control rats were treated with EG/AC but not lemon juice. Negative control rats were provided with normal drinking water, and were administered normal water by gavage. Each group contained 6 rats. After 10 days, serum samples were collected for analysis, the left kidney was removed and assessed for calcium levels using flame spectroscopy, and the right kidney was sectioned for histopathological analysis using light microscopy. RESULTS: Analysis showed that the rats treated with EG/AC alone had higher amounts of calcium in the kidneys compared to negative control rats. This EG/AC-induced increase in kidney calcium levels was inhibited by the administration of lemon juice. Histology showed that rats treated with EG/AC alone had large deposits of calcium oxalate crystals in all parts of the kidney, and that such deposits were not present in rats also treated with either 100% or 75% lemon juice. CONCLUSION: These data suggest that lemon juice has a protective activity against urolithiasis. PMID- 17919316 TI - Genotype-specific interactions and the trade-off between host and parasite fitness. AB - BACKGROUND: Evolution of parasite traits is inextricably linked to their hosts. For instance one common definition of parasite virulence is the reduction in host fitness due to infection. Thus, traits of infection must be viewed in both protagonists and may be under shared genetic and physiological control. We investigated these questions on the oomycete Hyaloperonospora arabidopsis (= parasitica), a natural pathogen of the Brassicaceae Arabidopsis thaliana. RESULTS: We performed a controlled cross inoculation experiment confronting six lines of the host plant with seven strains of the parasite in order to evaluate genetic variation for phenotypic traits of infection among hosts, parasites, and distinct combinations. Parasite infection intensity and transmission were highly variable among parasite strains and host lines but depended also on the interaction between particular genotypes of the protagonists, and genetic variation for the infection phenotype of parasites from natural populations was found even at a small spatial scale within population. Furthermore, increased parasite fitness led to a significant decrease in host fitness only on a single host line (Gb), although a trade-off between these two traits was expected because host and parasite share the same resource pool for their respective reproduction. We propose that different levels of compatibility dependent on genotype by genotype interactions might lead to different amounts of resources available for host and parasite reproduction. This variation in compatibility could thus mask the expected negative relationship between host and parasite fitness, as the total resource pool would not be constant. CONCLUSION: These results highlight the importance of host variation in the determination of parasite fitness traits. This kind of interaction may in turn decouple the relationship between parasite transmission and its negative effect on host fitness, altering theoretical predictions of parasite evolution. PMID- 17919317 TI - Disentangling the stigma of HIV/AIDS from the stigmas of drugs use, commercial sex and commercial blood donation - a factorial survey of medical students in China. AB - BACKGROUND: HIV/AIDS related stigma interferes with the provision of appropriate care and support for people living with HIV/AIDS. Currently, programs to address the stigma approach it as if it occurs in isolation, separate from the co-stigmas related to the various modes of disease transmission including injection drug use (IDU) and commercial sex (CS). In order to develop better programs to address HIV/AIDS related stigma, the inter-relationship (or 'layering') between HIV/AIDS stigma and the co-stigmas needs to be better understood. This paper describes an experimental study for disentangling the layering of HIV/AIDS related stigmas. METHODS: The study used a factorial survey design. 352 medical students from Guangzhou were presented with four random vignettes each describing a hypothetical male. The vignettes were identical except for the presence of a disease diagnosis (AIDS, leukaemia, or no disease) and a co-characteristic (IDU, CS, commercial blood donation (CBD), blood transfusion or no co-characteristic). After reading each vignette, participants completed a measure of social distance that assessed the level of stigmatising attitudes. RESULTS: Bivariate and multivariable analyses revealed statistically significant levels of stigma associated with AIDS, IDU, CS and CBD. The layering of stigma was explored using a recently developed technique. Strong interactions between the stigmas of AIDS and the co-characteristics were also found. AIDS was significantly less stigmatising than IDU or CS. Critically, the stigma of AIDS in combination with either the stigmas of IDU or CS was significantly less than the stigma of IDU alone or CS alone. CONCLUSION: The findings pose several surprising challenges to conventional beliefs about HIV/AIDS related stigma and stigma interventions that have focused exclusively on the disease stigma. Contrary to the belief that having a co-stigma would add to the intensity of stigma attached to people with HIV/AIDS, the findings indicate the presence of an illness might have a moderating effect on the stigma of certain co-characteristics like IDU. The strong interdependence between the stigmas of HIV/AIDS and the co-stigmas of IDU and CS suggest that reducing the co-stigmas should be an integral part of HIV/AIDS stigma intervention within this context. PMID- 17919318 TI - Modeling and detection of respiratory-related outbreak signatures. AB - BACKGROUND: Time series methods are commonly used to detect disease outbreak signatures (e.g., signals due to influenza outbreaks and anthrax attacks) from varying respiratory-related diagnostic or syndromic data sources. Typically this involves two components: (i) Using time series methods to model the baseline background distribution (the time series process that is assumed to contain no outbreak signatures), (ii) Detecting outbreak signatures using filter-based time series methods. METHODS: We consider time series models for chest radiograph data obtained from Midwest children's emergency departments. These models incorporate available covariate information such as patient visit counts and smoothed ambient temperature series, as well as time series dependencies on daily and weekly seasonal scales. Respiratory-related outbreak signature detection is based on filtering the one-step-ahead prediction errors obtained from the time series models for the respiratory-complaint background. RESULTS: Using simulation experiments based on a stochastic model for an anthrax attack, we illustrate the effect of the choice of filter and the statistical models upon radiograph attributed outbreak signature detection. CONCLUSION: We demonstrate the importance of using seasonal autoregressive integrated average time series models (SARIMA) with covariates in the modeling of respiratory-related time series data. We find some homogeneity in the time series models for the respiratory-complaint backgrounds across the Midwest emergency departments studied. Our simulations show that the balance between specificity, sensitivity, and timeliness to detect an outbreak signature differs by the emergency department and the choice of filter. The linear and exponential filters provide a good balance. PMID- 17919319 TI - qPIPSA: relating enzymatic kinetic parameters and interaction fields. AB - BACKGROUND: The simulation of metabolic networks in quantitative systems biology requires the assignment of enzymatic kinetic parameters. Experimentally determined values are often not available and therefore computational methods to estimate these parameters are needed. It is possible to use the three-dimensional structure of an enzyme to perform simulations of a reaction and derive kinetic parameters. However, this is computationally demanding and requires detailed knowledge of the enzyme mechanism. We have therefore sought to develop a general, simple and computationally efficient procedure to relate protein structural information to enzymatic kinetic parameters that allows consistency between the kinetic and structural information to be checked and estimation of kinetic constants for structurally and mechanistically similar enzymes. RESULTS: We describe qPIPSA: quantitative Protein Interaction Property Similarity Analysis. In this analysis, molecular interaction fields, for example, electrostatic potentials, are computed from the enzyme structures. Differences in molecular interaction fields between enzymes are then related to the ratios of their kinetic parameters. This procedure can be used to estimate unknown kinetic parameters when enzyme structural information is available and kinetic parameters have been measured for related enzymes or were obtained under different conditions. The detailed interaction of the enzyme with substrate or cofactors is not modeled and is assumed to be similar for all the proteins compared. The protein structure modeling protocol employed ensures that differences between models reflect genuine differences between the protein sequences, rather than random fluctuations in protein structure. CONCLUSION: Provided that the experimental conditions and the protein structural models refer to the same protein state or conformation, correlations between interaction fields and kinetic parameters can be established for sets of related enzymes. Outliers may arise due to variation in the importance of different contributions to the kinetic parameters, such as protein stability and conformational changes. The qPIPSA approach can assist in the validation as well as estimation of kinetic parameters, and provide insights into enzyme mechanism. PMID- 17919320 TI - Comparative genomics of Bacillus thuringiensis phage 0305phi8-36: defining patterns of descent in a novel ancient phage lineage. AB - BACKGROUND: The recently sequenced 218 kb genome of morphologically atypical Bacillus thuringiensis phage 0305phi8-36 exhibited only limited detectable homology to known bacteriophages. The only known relative of this phage is a string of phage-like genes called BtI1 in the chromosome of B. thuringiensis israelensis. The high degree of divergence and novelty of phage genomes pose challenges in how to describe the phage from its genomic sequences. RESULTS: Phage 0305phi8-36 and BtI1 are estimated to have diverged 2.0 - 2.5 billion years ago. Positionally biased Blast searches aligned 30 homologous structure or morphogenesis genes between 0305phi8-36 and BtI1 that have maintained the same gene order. Functional clustering of the genes helped identify additional gene functions. A conserved long tape measure gene indicates that a long tail is an evolutionarily stable property of this phage lineage. An unusual form of the tail chaperonin system split to two genes was characterized, as was a hyperplastic homologue of the T4gp27 hub gene. Within this region some segments were best described as encoding a conservative array of structure domains fused with a variable component of exchangeable domains. Other segments were best described as multigene units engaged in modular horizontal exchange. The non-structure genes of 0305phi8-36 appear to include the remnants of two replicative systems leading to the hypothesis that the genome plan was created by fusion of two ancestral viruses. The case for a member of the RNAi RNA-directed RNA polymerase family residing in 0305phi8-36 was strengthened by extending the hidden Markov model of this family. Finally, it was noted that prospective transcriptional promoters were distributed in a gradient of small to large transcripts starting from a fixed end of the genome. CONCLUSION: Genomic organization at a level higher than individual gene sequence comparison can be analyzed to aid in understanding large phage genomes. Methods of analysis include 1) applying a time scale, 2) augmenting blast scores with positional information, 3) categorizing genomic rearrangements into one of several processes with characteristic rates and outcomes, and 4) correlating apparent transcript sizes with genomic position, gene content, and promoter motifs. PMID- 17919321 TI - On human self-domestication, psychiatry, and eugenics. AB - The hypothesis that anatomically modern homo sapiens could have undergone changes akin to those observed in domesticated animals has been contemplated in the biological sciences for at least 150 years. The idea had already plagued philosophers such as Rousseau, who considered the civilization of man as going against human nature, and eventually "sparked over" to the medical sciences in the late 19th and early 20th century. At that time, human "self-domestication" appealed to psychiatry, because it served as a causal explanation for the alleged degeneration of the "erbgut" (genetic material) of entire populations and the presumed increase of mental disorders. Consequently, Social Darwinists emphasised preventing procreation by people of "lower genetic value" and positively selecting favourable traits in others. Both tendencies culminated in euthanasia and breeding programs ("Lebensborn") during the Nazi regime in Germany. Whether or not domestication actually plays a role in some anatomical changes since the late Pleistocene period is, from a biological standpoint, contentious, and the currently resurrected debate depends, in part, on the definitional criteria applied. However, the example of human self-domestication may illustrate that scientific ideas, especially when dealing with human biology, are prone to misuse, particularly if "is" is confused with "ought", i.e., if moral principles are deduced from biological facts. Although such naturalistic fallacies appear to be banned, modern genetics may, at least in theory, pose similar ethical problems to medicine, including psychiatry. In times during which studies into the genetics of psychiatric disorders are scientifically more valued than studies into environmental causation of disorders (which is currently the case), the prospects of genetic therapy may be tempting to alter the human genome in patients, probably at costs that no-one can foresee. In the case of "self domestication", it is proposed that human characteristics resembling domesticated traits in animals should be labelled "domestication-like", or better, objectively described as genuine adaptations to sedentism. PMID- 17919322 TI - Trinucleotide cassettes increase diversity of T7 phage-displayed peptide library. AB - BACKGROUND: Amino acid sequence diversity is introduced into a phage-displayed peptide library by randomizing library oligonucleotide DNA. We recently evaluated the diversity of peptide libraries displayed on T7 lytic phage and M13 filamentous phage and showed that T7 phage can display a more diverse amino acid sequence repertoire due to differing processes of viral morphogenesis. METHODS: In this study, we evaluated and compared the diversity of a 12-mer T7 phage displayed peptide library randomized using codon-corrected trinucleotide cassettes with a T7 and an M13 12-mer phage-displayed peptide library constructed using the degenerate codon randomization method. RESULTS: We herein demonstrate that the combination of trinucleotide cassette amino acid codon randomization and T7 phage display construction methods resulted in a significant enhancement to the functional diversity of a 12-mer peptide library. This novel library exhibited superior amino acid uniformity and order-of-magnitude increases in amino acid sequence diversity as compared to degenerate codon randomized peptide libraries. Comparative analyses of the biophysical characteristics of the 12-mer peptide libraries revealed the trinucleotide cassette-randomized library to be a unique resource. CONCLUSION: The combination of T7 phage display and trinucleotide cassette randomization resulted in a novel resource for the potential isolation of binding peptides for new and previously studied molecular targets. PMID- 17919323 TI - The progesterone receptor PROGINS polymorphism is not related to oxidative stress factors in women with polycystic ovary syndrome. AB - BACKGROUND: Women with PCOS have been reported to be at increased risk of a number of gynaecological neoplasias, including endometrial, breast, and ovarian cancer. Studies of the possible association of genetic variation in progesterone receptor polymorphism with risk of ovarian and breast cancer have concentrated on a variant known as PROGINS. METHODS: Ninety-five young women with PCOS and 99 healthy control women were included in our study. All subjects underwent venous blood drawing for complete hormonal assays, lipid profile, glucose, insulin and PROGINS polymorphism genetic study. RESULTS: In PROGINS polymorphism results; in both control and the patient groups T1/T1 has been detected in high levels. But for genotype (p = 0.178) and allele (p = 0.555) frequencies both of the groups give similar results. Statistically significant difference has been detected on serum FSH levels for T1/T1 genotype according to T2/T2 genotype. CONCLUSION: No relation has been detected between the inflammatory and oxidative stress factors, and PROGINS polymorphism alleles. This may be because the PCOS patients are young and their BMI means are normal and their CIMT and oxidative stress markers are like healthy women. PMID- 17919324 TI - Divergent evolution of arrested development in the dauer stage of Caenorhabditis elegans and the infective stage of Heterodera glycines. AB - BACKGROUND: The soybean cyst nematode Heterodera glycines is the most important parasite in soybean production worldwide. A comprehensive analysis of large-scale gene expression changes throughout the development of plant-parasitic nematodes has been lacking to date. RESULTS: We report an extensive genomic analysis of H. glycines, beginning with the generation of 20,100 expressed sequence tags (ESTs). In-depth analysis of these ESTs plus approximately 1,900 previously published sequences predicted 6,860 unique H. glycines genes and allowed a classification by function using InterProScan. Expression profiling of all 6,860 genes throughout the H. glycines life cycle was undertaken using the Affymetrix Soybean Genome Array GeneChip. Our data sets and results represent a comprehensive resource for molecular studies of H. glycines. Demonstrating the power of this resource, we were able to address whether arrested development in the Caenorhabditis elegans dauer larva and the H. glycines infective second-stage juvenile (J2) exhibits shared gene expression profiles. We determined that the gene expression profiles associated with the C. elegans dauer pathway are not uniformly conserved in H. glycines and that the expression profiles of genes for metabolic enzymes of C. elegans dauer larvae and H. glycines infective J2 are dissimilar. CONCLUSION: Our results indicate that hallmark gene expression patterns and metabolism features are not shared in the developmentally arrested life stages of C. elegans and H. glycines, suggesting that developmental arrest in these two nematode species has undergone more divergent evolution than previously thought and pointing to the need for detailed genomic analyses of individual parasite species. PMID- 17919325 TI - Effect of surgeon on transprosthetic gradients after aortic valve replacement with Freestyle stentless bioprosthesis and its consequences: a follow-up study in 587 patients. AB - BACKGROUND: The implantation of stentless valves is technically demanding and the outcome may depend on the performance of surgeons. We studied systematically the role of surgeons and other possible determinants for mid-term survival, postoperative gradients and Quality of Life (QoL) after aortic valve replacement (AVR) with Freestyle stentless bioprostheses. METHODS: Between 1996 and 2003, 587 patients (mean 75 years) underwent AVR with stentless Medtronic Freestyle(R) bioprostheses. Follow-up was 99% complete. Determinants of morbidity, mortality, survival time and QoL were evaluated by multiple, time-related, regression analysis. Risk models were built for all sections of the Nottingham Health Profile (NHP): energy, pain, emotional reaction, sleep, social isolation and physical mobility RESULTS: Actuarial freedom from aortic valve re-operation, structural valve deterioration, non-structural valve dysfunction, prosthetic valve endocarditis and thromboembolic events at 6 years were 95.9 +/- 2.1%, 100%, 98.7 +/- 0.5%, 97.0 +/- 1.5%, 79.6 +/- 4.3%, respectively. The actuarial freedom from bleeding events at 6 years was 93.1 +/- 1.9%. Estimated survival at 6 years was similar to the age-matched German population (61.4 +/- 3.8 %). Predictors of survival time were: diabetes mellitus, atrial fibrillation, peripheral vascular disease, renal dysfunction, female gender > 80 years and patients < 165 cm with BMI < 24. Predictive models showed characteristic profiles and good discriminative powers (c-indexes > 0.7) for each of the 6 QoL sections. Early transvalvular gradients were identified as independent risk factors for impaired physical mobility (c-index 0.77, p < 0.002). A saturated propensity score identified besides patient related factors (e.g. preoperative gradients, ejection fraction, haematological factors) indexed geometric orifice area, subcoronary implantation technique and individual surgeons as predictors of high gradients. CONCLUSION: In addition to the valve size (in relation to body size), subcoronary technique (versus total root) and various patient-related factors the risk of elevated gradients after stentless valve implantation depends, considerably on the individual surgeon. Although there was no effect on survival time and most aspects of QoL, higher postoperative transvalvular gradients affect physical mobility after AVR. PMID- 17919326 TI - The 2G allele of promoter region of matrix metalloproteinase-1 as an essential pre-condition for the early onset of oral squamous cell carcinoma. AB - BACKGROUND: Matrix metalloproteinase (MMP) is known to be involved in the initial and progressive stages of cancer development, and in the aggressive phenotypes of cancer. This study examines the association of single nucleotide polymorphisms in promoter regions of MMP-1 and MMP-3 with susceptibility to oral squamous cell carcinoma (OSCC). METHODS: We compared 170 Japanese OSCC cases and 164 healthy controls for genotypes of MMP-1 and MMP-3. RESULTS: The frequency of the MMP-1 2G allele was higher and that of the 1G homozygote was lower in the OSCC cases (p = 0.034). A multivariate logistic regression analysis revealed that subjects who were 45 years old or older had a significantly increased (2.47-fold) risk of OSCC (95%CI 1.47-4.14, p = 0.0006), and those carrying the MMP-1 2G allele had a 2.30 fold risk (95%CI 1.15-4.58, p = 0.018), indicating independent involvement of these factors in OSCC. One of the key discoveries of this research is the apparent reduction of the MMP-1 1G/1G and 1G/2G genotype distributions among the early onset OSCC cases under the ages of 45 years. It should be noted that the tongue was the primary site in 86.2% of these early onset cases. This could suggest the specific carcinogenic mechanisms, i.e. specific carcinogenic stimulations and/or genetic factors in the tongue. CONCLUSION: Since the 2G allele is a majority of the MMP-1 genotype in the general population, it seems to act as a genetic pre-condition in OSCC development. However this report suggests a crucial impact of the MMP-1 2G allele in the early onset OSCC. PMID- 17919327 TI - Acute effects of ingesting Java Fittrade mark energy extreme functional coffee on resting energy expenditure and hemodynamic responses in male and female coffee drinkers. AB - BACKGROUND: The purpose of this study was to examine the effects of a functional coffee beverage containing additional caffeine, green tea extracts, niacin and garcinia cambogia to regular coffee to determine the effects on resting energy expenditure (REE) and hemodynamic variables. METHODS: Subjects included five male (26 +/- 2.1 y, 97.16 +/- 10.05 kg, 183.89 +/- 6.60 cm) and five female (28.8 +/- 5.3 y, 142.2 +/- 12.6 lbs) regular coffee drinkers. Subjects fasted for 10 hours and were assessed for 1 hour prior (PRE) and 3 hours following 1.5 cups of coffee ingestion [JavaFittrade mark Energy Extreme (JF) ~400 mg total caffeine; Folgers (F) ~200 mg total caffeine] in a double-blind, crossover design. REE, resting heart rate (RHR), and systolic (SBP) and diastolic (DBP) blood pressure was assessed at PRE and 1, 2, and 3-hours post coffee ingestion. Data were analyzed by three-factor repeated measures ANOVA (p < 0.05). RESULTS: JF trial resulted in a significant main effect for REE (p < 0.01), SBP (p < 0.01), RER (p < 0.01), and VO2 (p < 0.01) compared to F, with no difference between trials on the RHR and DBP variables. A significant interaction for trial and time point (p < 0.05) was observed for the variable REE. The JF trial resulted in a significant overall mean increase in REE of 14.4% (males = 12.1%, females = 17.9%) over the observation period (p < 0.05), while the F trial produced an overall decrease in REE of 5.7%. SBP was significantly higher in the JF trial; however, there was no significant increase from PRE to 3-hours post. CONCLUSION: Results from this study suggest that JavaFittrade mark Energy Extreme coffee is more effective than Folgers regular caffeinated coffee at increasing REE in regular coffee drinkers for up to 3 hours following ingestion without any adverse hemodynamic effects. PMID- 17919328 TI - Host origin of plastid solute transporters in the first photosynthetic eukaryotes. AB - BACKGROUND: It is generally accepted that a single primary endosymbiosis in the Plantae (red, green (including land plants), and glaucophyte algae) common ancestor gave rise to the ancestral photosynthetic organelle (plastid). Plastid establishment necessitated many steps, including the transfer and activation of endosymbiont genes that were relocated to the nuclear genome of the 'host' followed by import of the encoded proteins into the organelle. These innovations are, however, highly complex and could not have driven the initial formation of the endosymbiosis. We postulate that the re-targeting of existing host solute transporters to the plastid fore-runner was critical for the early success of the primary endosymbiosis, allowing the host to harvest endosymbiont primary production. RESULTS: We tested this model of transporter evolution by conducting a comprehensive analysis of the plastid permeome in Arabidopsis thaliana. Of 137 well-annotated transporter proteins that were initially considered, 83 that are broadly distributed in Plantae were submitted to phylogenetic analysis. Consistent with our hypothesis, we find that 58% of Arabidopsis transporters, including all carbohydrate transporters, are of host origin, whereas only 12% arose from the cyanobacterial endosymbiont. Four transporter genes are derived from a Chlamydia-like source, suggesting that establishment of the primary plastid likely involved contributions from at least two prokaryotic sources. CONCLUSION: Our results indicate that the existing plastid solute transport system shared by Plantae is derived primarily from host genes. Important contributions also came from the cyanobacterial endosymbiont and Chlamydia-like bacteria likely co-resident in the first algae. PMID- 17919329 TI - Accuracy of telepsychiatric assessment of new routine outpatient referrals. AB - BACKGROUND: Studies on the feasibility of telepsychiatry tend to concentrate only on a subset of clinical parameters. In contrast, this study utilises data from a comprehensive assessment. The main objective of this study is to compare the accuracy of findings from telepsychiatry with those from face to face interviews. METHOD: This is a primary, cross-sectional, single-cluster, balanced crossover, blind study involving new routine psychiatric referrals. Thirty-seven out of forty cases fulfilling the selection criteria went through a complete set of independent face to face and video assessments by the researchers who were blind to each other's findings. RESULTS: The accuracy ratio of the pooled results for DSM-IV diagnoses, risk assessment, non-drug and drug interventions were all above 0.76, and the combined overall accuracy ratio was 0.81. There were substantial intermethod agreements for Cohen's kappa on all the major components of evaluation except on the Risk Assessment Scale where there was only weak agreement. CONCLUSION: Telepsychiatric assessment is a dependable method of assessment with a high degree of accuracy and substantial overall intermethod agreement when compared with standard face to face interview for new routine outpatient psychiatric referrals. PMID- 17919330 TI - Sexual behaviors and their correlates among young people in Mauritius: a cross sectional study. AB - BACKGROUND: Little is known about the HIV/AIDS epidemic in the Indian Ocean region, including Mauritius. National records suggest a prevalence of HIV in Mauritius of < 1% in the general population, which is one of the lowest prevalence rates in southern Africa. However, HIV-positive cases have been increasing recently in Mauritius. We conducted a cross-sectional survey in January 2003 to assess the prevalence of HIVrelated sexual behaviors and their correlates among young people aged 15-24 years in Mauritius. METHODS: We identified 1200 participants using two-stage cluster sampling. Demographic, social, sexual, and knowledge of HIV/AIDS data were obtained in face-to-face interviews using a structured questionnaire administered by trained interviewers. The prevalence of sexual behaviors was described in relation to gender, and the correlates of ever having had sex and nonuse of condom at last sex were analyzed using logistic regression. RESULTS: In the target population, 30.9% of males and 9.7% of females reported a history of sexual intercourse. Of the currently sexually active participants, 50.6% of men and 71.2% of women did not use condoms at their last sexual encounter. Logistic regression revealed that work experience and marijuana use were significantly associated with men's sexual experience, whereas being out of school and drinking experience were significantly associated with women's sexual experience. For both men and women, being Christian and visiting nightclubs were associated with having ever had sexual intercourse (P < 0.05). In addition, not using a condom at the first sexual encounter and lack of exposure to a nongovernmental organization (NGO) dealing with HIV/AIDS were associated with the nonuse of condoms at the last sexual encounter (P < 0.05). CONCLUSION: Young people in Mauritius are at risk of a future HIV epidemic because behaviors predisposing to HIV infection are prevalent among sexually experienced youth. A focused prevention program targeting young people should be reinforced as part of the National AIDS Control Program, taking into account the predictors of sexual behaviors identified here. PMID- 17919332 TI - Thalamic haemorrhage vs internal capsule-basal ganglia haemorrhage: clinical profile and predictors of in-hospital mortality. AB - BACKGROUND: There is a paucity of clinical studies focused specifically on intracerebral haemorrhages of subcortical topography, a subject matter of interest to clinicians involved in stroke management. This single centre, retrospective study was conducted with the following objectives: a) to describe the aetiological, clinical and prognostic characteristics of patients with thalamic haemorrhage as compared with that of patients with internal capsule basal ganglia haemorrhage, and b) to identify predictors of in-hospital mortality in patients with thalamic haemorrhage. METHODS: Forty-seven patients with thalamic haemorrhage were included in the "Sagrat Cor Hospital of Barcelona Stroke Registry" during a period of 17 years. Data from stroke patients are entered in the stroke registry following a standardized protocol with 161 items regarding demographics, risk factors, clinical features, laboratory and neuroimaging data, complications and outcome. The region of the intracranial haemorrhage was identified on computerized tomographic (CT) scans and/or magnetic resonance imaging (MRI) of the brain. RESULTS: Thalamic haemorrhage accounted for 1.4% of all cases of stroke (n = 3420) and 13% of intracerebral haemorrhage (n = 364). Hypertension (53.2%), vascular malformations (6.4%), haematological conditions (4.3%) and anticoagulation (2.1%) were the main causes of thalamic haemorrhage. In-hospital mortality was 19% (n = 9). Sensory deficit, speech disturbances and lacunar syndrome were significantly associated with thalamic haemorrhage, whereas altered consciousness (odds ratio [OR] = 39.56), intraventricular involvement (OR = 24.74) and age (OR = 1.23), were independent predictors of in-hospital mortality. CONCLUSION: One in 8 patients with acute intracerebral haemorrhage had a thalamic hematoma. Altered consciousness, intraventricular extension of the hematoma and advanced age were determinants of a poor early outcome. PMID- 17919331 TI - Statistical power of phylo-HMM for evolutionarily conserved element detection. AB - BACKGROUND: An important goal of comparative genomics is the identification of functional elements through conservation analysis. Phylo-HMM was recently introduced to detect conserved elements based on multiple genome alignments, but the method has not been rigorously evaluated. RESULTS: We report here a simulation study to investigate the power of phylo-HMM. We show that the power of the phylo-HMM approach depends on many factors, the most important being the number of species-specific genomes used and evolutionary distances between pairs of species. This finding is consistent with results reported by other groups for simpler comparative genomics models. In addition, the conservation ratio of conserved elements and the expected length of the conserved elements are also major factors. In contrast, the influence of the topology and the nucleotide substitution model are relatively minor factors. CONCLUSION: Our results provide for general guidelines on how to select the number of genomes and their evolutionary distance in comparative genomics studies, as well as the level of power we can expect under different parameter settings. PMID- 17919333 TI - Comparative genomic analysis of Campylobacter jejuni associated with Guillain Barre and Miller Fisher syndromes: neuropathogenic and enteritis-associated isolates can share high levels of genomic similarity. AB - BACKGROUND: Campylobacter jejuni infection represents the most frequent antecedent infection triggering the onset of the neuropathic disorders Guillain Barre syndrome (GBS) and Miller Fisher syndrome (MFS). Although sialylated ganglioside-mimicking lipo-oligosaccharide (LOS) structures are the strongest neuropathogenic determinants in C. jejuni, they do not appear to be the only requirement for a neuropathic outcome since strains capable of their production have been isolated from patients with uncomplicated cases of enteritis. Consequently, other pathogen and/or host-related factors contribute to the onset of neurological complications. We have used comparative genomic hybridization to perform a detailed genomic comparison of strains isolated from GBS/MFS and enteritis-only patients. Our dataset, in which the gene conservation profile for 1712 genes was assayed in 102 strains, including 56 neuropathogenic isolates, represents the largest systematic search for C. jejuni factors associated with GBS/MFS to date and has allowed us to analyze the genetic background of neuropathogenic C. jejuni strains with an unprecedented level of resolution. RESULTS: The majority of GBS/MFS strains can be assigned to one of six major lineages, suggesting that several genetic backgrounds can result in a neuropathogenic phenotype. A statistical analysis of gene conservation rates revealed that although genes involved in the sialylation of LOS structures were significantly associated with neuropathogenic strains, still many enteritis control strains both bear these genes and share remarkable levels of genomic similarity with their neuropathogenic counterparts. Two capsule biosynthesis genes (Cj1421c and Cj1428c) showed higher conservation rates among neuropathogenic strains compared to enteritis-control strains. Any potential involvement of these genes in neuropathogenesis must be assessed. A single gene (HS:3 Cj1135) had a higher conservation rate among enteritis-control strains. This gene encodes a glucosyltransferase that is found in some of the LOS classes that do not express ganglioside mimics. CONCLUSION: Our findings corroborate that neuropathogenic factors may be transferred between unrelated strains of different genetic background. Our results would also suggest that the failure of some strains isolated from uncomplicated cases of enteritis to elicit a neuropathic clinical outcome may be due to subtle genetic differences that silence their neuropathogenic potential and/or due to host-related factors. PMID- 17919334 TI - Use of formative research in developing a knowledge translation approach to rotavirus vaccine introduction in developing countries. AB - BACKGROUND: Rotavirus gastroenteritis is the leading cause of diarrheal disease mortality among children under five, resulting in 450,000 to 700,000 deaths each year, and another 2 million hospitalizations, mostly in the developing world. Nearly every child in the world is infected with rotavirus at least once before they are five years old. Vaccines to prevent rotavirus or minimize its severity are now becoming available, and have already been introduced into the public vaccine programs of several Latin American countries. The World Health Organization (WHO) has made rotavirus vaccine introduction in developing countries a high priority. The WHOs Guidelines for Vaccine Introduction indicates that a key determinant to achieving vaccine introduction is the public health priority of the disease, suggesting that where the disease is not a priority uptake of the vaccine is unlikely. WHO recommends conducting a qualitative analysis of opinions held by the public health community to determine the perceptions of the disease and the priority given to the vaccine. METHODS: This paper presents the formative research results of a qualitative survey of public health providers in five low- and middle-income countries to determine if and to what degree rotavirus is perceived to be a problem and the priority of a vaccine. Open-ended surveys were carried out through focus group discussions and one-on one interviews. RESULTS: Researchers discovered that in all five countries knowledge of rotavirus was extremely low, and as a result was not considered a high priority. However, diarrhea among young children was considered a high priority among public health providers in the three poorest countries with relatively high levels of child mortality: India, Indonesia, and Nicaragua. CONCLUSION: In the poorest countries, advocacy and communication efforts to raise awareness about rotavirus sufficient for prioritization and accelerated vaccine introduction might benefit from a knowledge translation approach that delivers information and evidence about rotavirus through the broader context of diarrheal disease control, an existing priority, and including information about other new interventions, specifically low-osmolarity oral rehydration solution and zinc treatment. PMID- 17919335 TI - Policy maker and health care provider perspectives on reproductive decision making amongst HIV-infected individuals in South Africa. AB - BACKGROUND: Worldwide there is growing attention paid to the reproductive decisions faced by HIV-infected individuals. Studies in both developed and developing countries have suggested that many HIV-infected women continue to desire children despite knowledge of their HIV status. Despite the increasing attention to the health care needs of HIV-infected individuals in low resource settings, little attention has been given to reproductive choice and intentions. Health care providers play a crucial role in determining access to reproductive health services and their influence is likely to be heightened in delivering services to HIV-infected women. We examined the attitudes of health care policy makers and providers towards reproductive decision-making among HIV-infected individuals. METHODS: In-depth interviews were conducted with 14 health care providers at two public sector health care facilities located in Cape Town, South Africa. In addition, 12 in-depth interviews with public sector policy makers and managers, and managers within HIV/AIDS and reproductive health NGOs were conducted. Data were analyzed using a grounded theory approach. RESULTS: Providers and policy makers approached the issues related to being HIV-infected and child bearing differently. Biomedical considerations were paramount in providers' approaches to HIV infection and reproductive decision-making, whereas, policy makers approached the issues more broadly recognizing the structural constraints that inform the provision of reproductive health care services and the possibility of "choice" for HIV-infected individuals. CONCLUSION: The findings highlight the diversity of perspectives among policy makers and providers regarding the reproductive decisions taken by HIV-infected people. There is a clear need for more explicit policies recognizing the reproductive rights and choices of HIV-infected individuals. PMID- 17919336 TI - What orthopaedic surgery residents need to know about the hand and wrist? AB - BACKGROUND: To develop a Core Curriculum for Orthopaedic Surgery; and to conduct a national survey to assess the importance of curriculum items as judged by orthopaedic surgeons with primary affiliation non-academic. Attention for this manuscript was focused on determining the importance of topics pertaining to adult hand and wrist reconstruction. METHODS: A 281-item questionnaire was developed and consisted of three sections: 1) Validated Musculoskeletal Core Curriculum; 2) Royal College of Physician and Surgeons of Canada (RCPSC) Specialty Objectives and; 3) A procedure list. A random group of 131 [out of 156] orthopaedic surgeons completed the questionnaire. Data were analyzed descriptively and quantitatively using histograms, a Modified Hotel ling's T2 statistic 1 with p-value determined by a permutation test, and the Benjamini Hochberg/Yekutieli procedure RESULTS: 131/156 (84%) orthopaedic surgeons participated in this study. 27/32 items received an average mean score of at least 3.0/4.0 by all respondents thus suggesting that 84% of the items are either "probably important" or "important" to know by the end of residency (SD range 0.007-0.228). The Benjamini-Hochberg procedure demonstrated that for 80% of the 32 x 31/2 = 496 possible pairs of hand and wrist questions did not appear to demonstrate the same distribution of ratings given that one question was different from that of another question. CONCLUSION: This study demonstrates with reliable statistical evidence, agreement on the importance of 27/32 items pertaining to hand and wrist reconstruction is included in a Core Curriculum for Orthopaedic Surgery. Residency training programs need ensure that educational opportunities focusing on the ability to perform with proficiency procedures pertaining to the hand and wrist is taught and evaluated in their respective programs. PMID- 17919337 TI - The radiosensitizing effect of doranidazole on human colorectal cancer cells exposed to high doses of irradiation. AB - BACKGROUND: This paper investigates the effects of a new radiosensitizer, doranidazole, and enhancing irradiation on colorectal cancer cells. METHODS: The radiosensitizing effect of doranidazole was determined using colony formation and propidium iodide (PI) assays to measure cell growth inhibition and the cell killing effect of human colorectal cancer cell lines exposed to high doses of gamma-ray irradiation under hypoxic conditions in vitro. Fluorescence staining and cell migration assays were also used to assess the radiosensitizing effect. RESULTS: Cell proliferation evaluated by clonogenic survival curves was significantly inhibited by 5 mmol/L doranidazole, particularly at doses ranging from 10 to 30 Gy of irradiation. The radiosensitizing effect of doranidazole on colorectal cancer cells occurs in a time- and dose-dependent manner. Doranidazole also inhibited the mobility of cell invasion and migration. CONCLUSION: Doranidazole can enhance the killing effect and the cell growth inhibition of colorectal cancer after high-dose irradiation in a time and dose-dependent manner. PMID- 17919338 TI - Docetaxel weekly regimen in conjunction with RF hyperthermia for pretreated locally advanced non-small cell lung cancer: a preliminary study. AB - BACKGROUND: To evaluate the feasibility and therapeutic effect of chemotherapy combined with regional radio frequency hyperthermia for pretreated locally advanced non-small cell lung cancer. METHODS: 29 patients with stage III non small cell lung cancer were enrolled in present study, received chemotherapy up to 4 cycles and radio frequency hyperthermia up to 32 times. The primary end points were grade 3,4 hematological or non-hematological toxicities and progression free survival, the secondary end points were response rate, tumor control rate and overall survival. Method of Kaplan-Meier was used for the survival analysis. RESULTS: 21 patients completed their arranged treatments. The most common grade 3,4 toxicity was neutropenia (24.1%). Median progression free survival was 4 months (range 0-13 months), one year progression free survival rate was 10.3%. Overall response rate was 25.9%, tumor control rate was 66.6%. Median overall survival was 11 months (range 2-18+ months), one year overall survival rate was 44.8%. CONCLUSION: Treatment of chemotherapy in conjunction with regional hyperthermia was safe and well tolerant, it suggested an impressive tumor control rate and an acceptable one year progression free survival. Further study might be needed. PMID- 17919339 TI - Relationships between sensory stimuli and autonomic nervous regulation during real and virtual exercises. AB - BACKGROUND: Application of virtual environment (VE) technology to motor rehabilitation increases the number of possible rehabilitation tasks and/or exercises. However, enhancing a specific sensory stimulus sometimes causes unpleasant sensations or fatigue, which would in turn decrease motivation for continuous rehabilitation. To select appropriate tasks and/or exercises for individuals, evaluation of physical activity during recovery is necessary, particularly the changes in the relationship between autonomic nervous activity (ANA) and sensory stimuli. METHODS: We estimated the ANA from the R-R interval time series of electrocardiogram and incoming sensory stimuli that would activate the ANA. For experiments in real exercise, we measured vehicle data and electromyogram signals during cycling exercise. For experiments in virtual exercise, we measured eye movement in relation to image motion vectors while the subject was viewing a mountain-bike video image from a first-person viewpoint. RESULTS: For the real cycling exercise, the results were categorized into four groups by evaluating muscle fatigue in relation to the ANA. They suggested that fatigue should be evaluated on the basis of not only muscle activity but also autonomic nervous regulation after exercise. For the virtual exercise, the ANA related conditions revealed a remarkable time distribution of trigger points that would change eye movement and evoke unpleasant sensations. CONCLUSION: For expanding the options of motor rehabilitation using VE technology, approaches need to be developed for simultaneously monitoring and separately evaluating the activation of autonomic nervous regulation in relation to neuromuscular and sensory systems with different time scales. PMID- 17919341 TI - Breastfeeding, breast milk and viruses. AB - BACKGROUND: There is seemingly consistent and compelling evidence that there is no association between breastfeeding and breast cancer. An assumption follows that milk borne viruses cannot be associated with human breast cancer. We challenge this evidence because past breastfeeding studies did not determine "exposure" of newborn infants to colostrum and breast milk. METHODS: We conducted a prospective review of 100 consecutive births of infants in the same centre to determine the proportion of newborn infants who were "exposed" to colostrum or breast milk, as distinct from being fully breast fed. We also report a review of the breastfeeding practices of mothers of over 87,000 newborn infants in the Australian State of New South Wales. This study was approved by the Human Research Ethics Committee of the University of New South Wales (Sydney, Australia). Approval 05063, 29 September 2005. RESULTS: Virtually all (97 of 100) newborn infants in this centre were "exposed" to colostrum or breast milk whether or not they were fully breast fed. Between 82.2% to 98.7% of 87,000 newborn infants were "exposed" to colostrum or breast milk. CONCLUSION: In some Western communities there is near universal exposure of new born infants to colostrum and breast milk. Accordingly it is possible for the transmission of human milk borne viruses. This is contrary to the widespread assumption that human milk borne viruses cannot be associated with breast cancer. PMID- 17919340 TI - Characterization of a novel bifunctional mutant of staphylokinase with platelet targeted thrombolysis and antiplatelet aggregation activities. AB - BACKGROUND: Although staphylokianse (SAK) is among the most promising blood dissolving agents, it is far from ideal. It is interesting to hypothesize that the clot lysis efficacy of SAK can be enhanced with direct active platelet binding ability, and at the same time the rethrombosis complication after successful recanalization can be minimized with an antiplatelet aggregation activity. The present study was performed to characterize the functional properties of RGD-SAK, a novel mutant of staphylokinase (SAK). RESULTS: By using site-directed mutagenesis, an Arg-Gly-Asp (RGD) motif was engineered in the staphylokinase (SAK). This mutant of SAK designated RGD-SAK was expressed, purified and characterized. Biochemical analysis indicated that RGD-SAK maintained the similar structure and the fibrinolytic function of SAK. Measurement of platelet binding activity in vitro demonstrated that RGD-SAK had a much higher affinity with platelets than SAK. In vitro platelet-rich clot lysis assay demonstrated that the engineered mutant outperformed the non-manipulated SAK. The time required for 50% platelet-rich clot lysis and the concentration required to obtain 50% clot lysis (C50) were reduced significantly across different concentrations of RGD-SAK comparing with SAK. Meanwhile, RGD-SAK was found to inhibit ADP-induced platelet aggregation in a concentration-dependent manner while SAK had negligible effect on platelet aggregation. CONCLUSION: RGD SAK possessed the bifunction to target platelet-rich clots and to block platelets aggregation, and thus may serve as a more potential thrombolytic agent with platelet-targeted thrombolytic and antiplatelet aggregation activities in compared with SAK. PMID- 17919342 TI - Improving the teaching skills of residents as tutors/facilitators and addressing the shortage of faculty facilitators for PBL modules. AB - BACKGROUND: Residents play an important role in teaching of medical undergraduate students. Despite their importance in teaching undergraduates they are not involved in any formal training in teaching and leadership skills. We aimed to compare the teaching skills of residents with faculty in facilitating small group Problem Based Learning (PBL) sessions. METHODS: This quasi experimental descriptive comparative research involved 5 postgraduate year 4 residents and five senior faculty members. The study was conducted with all phase III (Final year) students rotating in Gastroenterology. The residents and faculty members received brief training of one month in facilitation and core principles of adult education. Different aspects of teaching skills of residents and faculty were evaluated by students on a questionnaire (graded on Likert Scale from 1 to 10) assessing i) Knowledge Base-content Learning (KBL), ii) PBL, iii) Student Centered Learning (SCL) and iv) Group Skills (GS). RESULTS: There were 33 PBL teaching sessions in which 120 evaluation forms were filled; out of these 53% forms were filled for residents and 47% for faculty group. The faculty showed a statistically greater rating in "KBL" (faculty 8.37 Vs resident 7.94; p-value 0.02), "GS" (faculty 8.06 vs. residents 7.68; p-value 0.04). Differences in faculty and resident scores in "the PBL" and "SCL" were not significant. The overall score of faculty facilitators, however, was statistically significant for resident facilitators. (p = .05). CONCLUSION: 1) Residents are an effective supplement to faculty members for PBL; 2) Additional facilitators for PBL sessions can be identified in an institution by involvement of residents in teacher training workshops. PMID- 17919344 TI - n-3 Fatty acids and mood: the devil is in the detail. PMID- 17919343 TI - Mitochondrial respiratory chain is involved in insulin-stimulated hydrogen peroxide production and plays an integral role in insulin receptor autophosphorylation in neurons. AB - BACKGROUND: Accumulated evidence suggests that hydrogen peroxide (H2O2) generated in cells during insulin stimulation plays an integral role in insulin receptor signal transduction. The role of insulin-induced H2O2 in neuronal insulin receptor activation and the origin of insulin-induced H2O2 in neurons remain unclear. The aim of the present study is to test the following hypotheses (1) whether insulin-induced H2O2 is required for insulin receptor autophosphorylation in neurons, and (2) whether mitochondrial respiratory chain is involved in insulin-stimulated H2O2 production, thus playing an integral role in insulin receptor autophosphorylation in neurons. RESULTS: Insulin stimulation elicited rapid insulin receptor autophosphorylation accompanied by an increase in H2O2 release from cultured cerebellar granule neurons (CGN). N-acetylcysteine (NAC), a H2O2 scavenger, inhibited both insulin-stimulated H2O2 release and insulin stimulated autophosphorylation of insulin receptor. Inhibitors of respiratory chain-mediated H2O2 production, malonate and carbonyl cyanide-4 (trifluoromethoxy)-phenylhydrazone (FCCP), inhibited both insulin-stimulated H2O2 release from neurons and insulin-stimulated autophosphorylation of insulin receptor. Dicholine salt of succinic acid, a respiratory substrate, significantly enhanced the effect of suboptimal insulin concentration on the insulin receptor autophosphorylation in CGN. CONCLUSION: Results of the present study suggest that insulin-induced H2O2 is required for the enhancement of insulin receptor autophosphorylation in neurons. The mitochondrial respiratory chain is involved in insulin-stimulated H2O2 production, thus playing an integral role in the insulin receptor autophosphorylation in neurons. PMID- 17919345 TI - Exogenous nucleosides accelerate differentiation of rat intestinal epithelial cells. AB - Several studies have reported differing data on the effect of exogenous nucleosides and nucleotides on the proliferation and differentiation in various intestinal cell lines and explants. To study whether exogenous nucleosides modulate intestinal cell differentiation, IEC-6 cells were differentiated in the presence or absence of a nucleoside mixture (cytidine, uridine, guanosine and inosine, 30 microM each), and the concentrations of nucleoside derivatives were determined by HPLC. Cell differentiation was assessed by electron microscopy, alkaline phosphatase activity and Rnd3 gene expression. The present results showed that uridine, guanosine and inosine were cleared from culture media (up to 32, 63 and 100 % in proliferating cells, and 31, 80 and 94 % in differentiated cells, respectively) whereas cytidine concentrations increased. Differentiation of IEC-6 cells was associated with a significant increase in intracellular nucleotide concentrations. Clearance of nucleosides correlated with a significant increase in the intracellular nucleotide pool in proliferating and differentiated IEC-6 cells. Intracellular guanosine nucleotides increased 2.5- and 5-fold in nucleoside-supplemented proliferating and differentiated cells, respectively. At 24 h, nucleoside-supplemented differentiated IEC-6 cells had significantly higher energy charge and GTP levels than non-supplemented ones. These modifications paralleled changes in cell differentiation as indicated by increased alkaline phosphatase activity, prolonged microvilli formation and accelerated down regulation of Rnd3 gene expression. The present findings suggest that exogenous nucleosides were selectively taken up by IEC-6 cells, increased the intracellular nucleotide pool, GTP and energy charge, and favoured cell morphological and functional changes during differentiation. PMID- 17919346 TI - Wholegrain foods made from a novel high-amylose barley variety (Himalaya 292) improve indices of bowel health in human subjects. AB - Himalaya 292 (Hordeum vulgare var. Himalaya 292) is a novel hull-less barley variety lacking activity of a key enzyme of starch synthesis giving a grain containing less total starch, more amylose and higher total dietary fibre. Animal trials have shown that Himalaya 292 contains more resistant starch and has greater positive impact on biomarkers of large-bowel health than comparable wholegrain cereal products. The present study compared the effects of foods made from wholegrain Himalaya 292 with those made from wholegrain wheat on faecal biomarkers of bowel health in human subjects. Seventeen male and female volunteers aged 31-66 years consumed similar quantities of Himalaya 292, whole wheat or refined cereal foods daily for 4 weeks in a randomised cross-over design. Total dietary fibre intakes from weighed food records were 45, 32 and 21 g/d for the Himalaya 292, whole-wheat and refined cereal periods, respectively. Compared with the refined cereal foods, consumption of Himalaya 292 foods resulted in 33 % higher faecal weight, a lowering of faecal pH from 7.24 to 6.98, a 42 % higher faecal concentration and a 91 % higher excretion of butyrate, a 57 % higher faecal total SCFA excretion and a 33 % lower faecal p-cresol concentration. pH and butyrate concentration and excretion were also significantly different compared with wholemeal wheat. It is concluded that consumption of a diet that included foods made from Himalaya 292 supplied more fibre and improved indices of bowel health compared with refined cereal foods and, for some indices, similar wholemeal wheat foods at equivalent levels of intake. PMID- 17919347 TI - Ductal carcinoma in situ: biology, diagnosis, and new therapies. AB - The incidence of ductal carcinoma in situ (DCIS) has markedly increased as a result of the use of screening mammography. Whether DCIS is a premalignant lesion or a cancer remains a cause of debate, but evidence supports the idea that DCIS evolves into invasive breast cancer based on histologic patterns, similar risk factors, and genetic similarities. Microcalcifications identified during mammography generally raise the suspicion of DCIS, and biopsy, often by core needle, confirms such a diagnosis. The extent of disease can be further delineated by breast magnetic resonance imaging. Radiation therapy in breast conserving treatment, along with tamoxifen, decreases the overall rate of local recurrence in patients with DCIS. Studies in the treatment of DCIS exploring partial breast radiation and trastuzumab are under way. Ongoing investigations with comparative genomic hybridization suggest that there are independent, evolutionary genetic pathways within DCIS. Genome-wide microarray-based gene expression analyses are now providing new opportunities to discover genes that are specifically activated or inactivated during the course of breast cancer progression. PMID- 17919348 TI - Patterns of bisphosphonate use in the United States in the treatment of metastatic bone disease. AB - PURPOSE: The purpose of this study was to determine the incidence of metastatic bone disease (MBD), the frequency of intravenous (I.V.) bisphosphonate use and its impact on skeletal-related events (SREs), and opioid use. PATIENTS AND METHODS: Patients diagnosed with MBD between 2000 and 2004 were identified using 2 Thomson MarketScan Research Databases. A total of 6783 patients, 1431 with breast cancer, fulfilled the criteria. Pain was assessed as the number of days on opioids, the strength of which was categorized according to the World Health Organization 3-step ladder for pain. RESULTS: Use of I.V. bisphosphonates steadily increased for all cancers from 17% in 2000 to 38% in 2004. For all patients, 61% received mild opioids and 35% received moderate to severe opioids at baseline. Use of I.V. bisphosphonates within the first 90 days after diagnosis of MBD was associated with a 63% decrease in SREs and reduction in use of moderate to severe opioids. Among patients with breast cancer, 10.6% received oral bisphosphonates before diagnosis of MBD, and 33.8% had pain at baseline. There was a 5.4% reduction in the use of moderate to severe opioids. CONCLUSION: Our data support the role of I.V. bisphosphonates in decreasing SRE and improving the quality of life for patients with MBD; the result is less pain and fewer SREs. More than 73% of all patients and 53% of patients with breast cancer never receive I.V. bisphosphonate treatment. Educational measures are warranted to increase the awareness by patients and physicians of the value of I.V. bisphosphonates in MBD. PMID- 17919349 TI - Feasibility and cardiac safety of pegylated liposomal doxorubicin plus trastuzumab in heavily pretreated patients with recurrent HER2-overexpressing metastatic breast cancer. AB - BACKGROUND: Few studies have evaluated concomitant pegylated liposomal doxorubicin (PLD) plus trastuzumab as therapy for HER2-overexpressing metastatic breast cancer (MBC). This open-label, prospective, phase II trial assessed the safety and efficacy of this regimen, with cardiac tolerance as the principal focus. PATIENTS AND METHODS: Women with HER2-overexpressing recurrent MBC, baseline left ventricular ejection fraction >or= 55%, and no history of serious cardiac illness were eligible; preexisting cardiac risk factors, including previous anthracyclines and previous trastuzumab for MBC, were allowed. Patients received weekly trastuzumab and every-3-week PLD until progression, prohibitive toxicity, or patient refusal. Left ventricular ejection fraction was assessed during and after therapy. Grade 3/4 congestive heart failure (CHF) was monitored for premature closure. RESULTS: The trial closed after 2.5 years for slow accrual. Twelve patients were enrolled: 7 had received adjuvant anthracyclines; 9 had received previous MBC treatment, of whom 7 had received trastuzumab in combination with chemotherapy. Patients received a mean of 4.8 cycles of PLD; 8 patients experienced stable disease; 4 patients experienced progression. Mean left ventricular ejection fraction levels did not change substantially: 60.4%, 57%, 60.3%, and 56.8% at baseline, after cycle 2, after cycle 4, and after completion of treatment, respectively. No patients experienced grade 4 CHF. One patient discontinued treatment after grade 3 CHF. Three patients experienced grade 2 left ventricular dysfunction, of whom 2 discontinued treatment. Cardiac function improved in all 4 patients after going off study. Other adverse events were generally mild (grade 1/2) and infrequent. CONCLUSION: Pegylated liposomal doxorubicin plus trastuzumab might be an option for heavily pretreated patients with recurrent HER2-overexpressing MBC. PMID- 17919351 TI - Method of detection of new contralateral primary breast cancer in younger versus older women. AB - BACKGROUND: Surveillance for contralateral primary breast cancer after a diagnosis of unilateral breast cancer typically consists of yearly mammography and physical examinations at 3-6 month intervals. Mammography is known to be less sensitive in younger, dense breasts. It is unknown at this time how well mammography performs in young patients to detect a new contralateral primary breast cancer. PATIENTS AND METHODS: Patients with contralateral breast cancer diagnosed between 1980 and 2004 were identified from the British Columbia Cancer Agency's Breast Cancer Outcomes Unit database in Vancouver. Characteristics of the tumor at baseline and the contralateral tumor were recorded as well as the method of detection of the contralateral breast cancer. A subset of patients was identified based on the age at which they were diagnosed with their initial primary cancer: < 40 years (group A) and 55-59 years (group B). chi2 and independent-sample t tests were used for between-group comparisons. RESULTS: Older patients were significantly more likely to have their second primary tumor detected by routine follow-up mammography compared with the younger cohort (P < 0.001). Older patients were also more likely to have estrogen receptor-positive, lower grade second primary tumors, and there was a trend toward smaller tumors. Tumors detected by mammography were more likely to be lower grade, estrogen receptor positive, and smaller. CONCLUSION: Older patients were more likely to have a contralateral breast cancer detected by conventional mammography, whereas younger patients tended to have cancer detected by physical examinations or by self-diagnosis. Better imaging techniques are required to detect new contralateral primary breast cancer in younger patients. PMID- 17919350 TI - Toxicity of dose-dense docetaxel followed by doxorubicin with cyclophosphamide as adjuvant therapy for breast cancer in a phase II study. AB - PURPOSE: In order to evaluate the feasibility of dose-dense docetaxel followed by dose-dense AC (doxorubicin/cyclophosphamide) as adjuvant chemotherapy for operable breast cancer, we conducted a phase II study. PATIENTS AND METHODS: In cohort 1, 28 patients received docetaxel 100 mg/m2 followed by doxorubicin 60 mg/m2 with cyclophosphamide 600 mg/m2, each every 2 weeks for 4 weeks (total of 8 cycles). Enrollment was discontinued because of stopping criteria based on significant toxicity (grade 4 hematologic toxicity or grade >or= 3 nonhematologic toxicity). In cohort 2, the docetaxel dose was reduced to 75 mg/m2; enrollment was discontinued after 18 patients. RESULTS: Significant toxicity occurred in 79% and 72% of patients in cohorts 1 and 2, respectively, resulting in treatment delays in 50% and 17% of patients, respectively. The most common grade 4 hematologic toxicity was neutropenia, which occurred in 7% and 42% of cohort 1 patients during docetaxel and AC, respectively, and in none and 19% of cohort 2 patients, respectively. The most common grade >or= 3 nonhematologic toxicity was palmar-plantar erythrodysesthesia, which occurred in 25% and none of cohort 1 patients during docetaxel and AC, respectively. With docetaxel 75 mg/m2 and patient education encouraging routine use of topical strategies, grade 3 palmar plantar erythrodysesthesia occurred in only 11% of cohort 2 patients. Grade 2 nail changes were also debilitating and occurred in 33% of cohort 1 patients during AC. CONCLUSION: These phase II findings suggest that dose-dense docetaxel 100 mg/m2 followed by AC is not feasible and, until more studies are conducted, should be restricted to clinical studies. PMID- 17919352 TI - Bilateral synchronous ductal carcinoma in situ in a young man: case report and review of the literature. AB - We report a rare case of synchronous bilateral ductal carcinoma in situ (DCIS) developing in a man with long-standing gynecomastia. He underwent bilateral staged subcutaneous mastectomies with the right side being performed first at age 26 years. Histology confirmed bilateral pure DCIS. There was no identifiable causative factor for the development of bilateral DCIS, and there was no familial history of the disease. He ultimately experienced progression to total bilateral mastectomies. This case highlights the importance of remaining vigilant about the presence of malignancy in normally benign conditions. PMID- 17919353 TI - Pure squamous cell carcinoma of the breast presenting as a pyogenic abscess: a case report. AB - The field of oncology is studded with fascinating case reports of rarities, and management of breast cancer by the oncologist has, at times, resulted in the surfacing of such instances of rarities. Pure squamous cell carcinoma (SCC) of the breast is such an example of a rare and generally aggressive malignancy constituting < 0.1% of invasive breast cancers. To the best of our knowledge, until 2006, only 5 patients of primary SCC of the breast, which presented clinically as breast abscess, have been reported in medical literature. We report the sixth worldwide case of pure primary SCC of the breast presenting as an abscess. In this report, we highlight the fact that a benign lesion like breast abscess can harbor such a rare malignancy. Clinicians should be aware of that fact, and adequate investigations should be done to rule out that possibility. Extensive literature review has been done to discuss the clinical and radiologic features as well as management of this rare lesion. PMID- 17919354 TI - Remission of porphyria cutanea tarda after anastrozole treatment of breast cancer. AB - Porphyria cutanea tarda (PCT) is sometimes precipitated or aggravated by increased exposure to estrogen, estrogen-like compounds, or tamoxifen. We report the case of a 37-year-old white woman who developed sporadic PCT after she took oral contraceptives for 10 years. She was heterozygous for the common H63D mutation of the hemochromatosis-associated HFE gene. The PCT responded partially to the cessation of oral contraceptives and to phlebotomy therapy to maintain low iron stores, but only remitted after she received anastrozole therapy for management of adenocarcinoma of the breast at age 59 years. The pertinence of HFE mutations, anastrozole and tamoxifen treatment, and chemotherapy to the development and management of PCT in women with breast cancer is discussed. PMID- 17919355 TI - Post-traumatic growth in advanced cancer patients receiving palliative care. AB - GOALS OF WORK: To develop the Greek version of the Post-traumatic Growth Inventory (PTGI-Gr), and assess its psychometric properties in a palliative care patient sample. PATIENTS AND METHODS: The scale was translated with the forward backward procedure to Greek. It was administered twice, with a 3-day interval, to 131 eligible patients with advanced cancer. Together with the PTGI, the patients also completed the Greek version of the Impact of Events Scale-Revised scale (IES R-Gr). The reliability was assessed by the internal consistency (Cronbach's alpha coefficients), test/retest (Spearman's r value), and inter-item correlations. Validity was demonstrated by factor analysis, inter-scale correlations, construct validity with the IES-R-Gr, and combined with the Eastern Cooperative Oncology Group (ECOG) performance status. MAIN RESULTS: The PTGI-Gr yielded a five-factor structure, explaining 73.5% of the variance. Cronbach's alphas for the five factors ranged from .66 to .87, respectively. Overall test-retest reliability was satisfactory with a range between .85 and .92 (p<.0005), and inter-item correlations ranged between .47 and .63. Inter-scale correlations were found satisfactory (p<.0005, p<.005, and p<.05). Validity as performed using combined validity analysis showed good results. Satisfactory construct validity was supported by the correlation analysis between the PTGI-Gr and the IES-R-Gr scales. CONCLUSIONS: PTGI-Gr is an instrument with satisfactory psychometric properties, and is a valid research tool for the post-traumatic growth of advanced cancer patients. PMID- 17919356 TI - Drug treatment discontinuation and achievement of target blood pressure and cholesterol in United Kingdom primary care. AB - AIM: This cohort study evaluated medication discontinuation and target achievement in a United Kingdom primary care setting. METHODS: The study population comprised patients within the General Practice Research Database who began treatment for hypertension, dyslipidaemia or both between 1997 and 2001. Discontinuation (absence of prescription refill > or = 6 months) and reaching treatment targets (blood pressure < 140/90 mmHg, or < 130/80 mmHg in patients with diabetes; total cholesterol < 193 mg/dL [5 mmol/L] or low-density lipoprotein cholesterol < 116 mg/dL [3 mmol/L]) were determined for patients treated for hypertension alone (cohort HT), dyslipidaemia alone (cohort DYS) and both conditions (cohort HT+DYS). RESULTS: At 2 years, 41.3% (95% CI: 40.8, 41.9%) of patients had stopped treatment in cohort HT, 29.2% (27.6, 30.9%) in cohort DYS and 25.0% (24.3, 25.8%) stopped either treatment in cohort HT+DYS. The cumulative proportion reaching treatment targets at this time was 28.2% (27.8, 28.7%) in cohort HT and 49.9% (47.8, 51.9%) in cohort DYS; in cohort HT+DYS, 43.4% (42.6, 44.2%) achieved blood pressure target, 53.7% (52.8, 54.6%) cholesterol target and 24.8% (24.0, 25.5%) reached target for both. Diabetic patients generally stayed on treatment longer and were less likely to attain their blood pressure targets, and more likely to reach cholesterol targets, compared with all patients. CONCLUSION: Patients prescribed both antihypertensive and lipid-lowering therapy remained on treatment longer; more patients achieved treatment target than those treated for single risk factors. Nevertheless, there is a large unmet need for initiating and maintaining antihypertensive and lipid-lowering therapy in patients with hypertension and dyslipidaemia. PMID- 17919357 TI - Comparison between spinal surgery blood transfusion services costs and associated treatment practices in the United States and Belgium. AB - OBJECTIVE: This study assessed utilization and cost of allogeneic blood transfusion (ABT) associated with spinal surgery in the United States (US) and Belgium. METHODS: A retrospective cohort of 292,864 spinal surgery inpatients in US hospitals was pooled with a cohort of 27,952 inpatients who had similar procedures in Belgian hospitals. Utilization and cost data were derived from hospital accounting systems. Costs were converted to US dollars. Descriptive and multivariate statistics were used to describe the factors associated with the use and cost of ABT. Missing data, confounding, and variable measurement error were addressed using standard approaches for observational studies. RESULTS: US hospitalizations cost $12,044 (SD = 15,920) over 3.6 days compared to $4010 (SD = 3586) over 10.3 days in Belgium. Low molecular weight heparin was used by 78% of Belgian patients and 4% of US patients. Red blood cell utilization occurred in approximately 7% of patients from both countries; however US patients received 6 units compared to 3 units by Belgian patients. US patients spent 3.5 (p < 0.0001) less days in hospital, 1.0 (p < 0.0001) more days in an intensive care unit, used 64% more allogeneic blood (OR = 1.64, 95% CI 1.53-1.75), and incurred $13,647 (p < 0.0001) more per hospitalization than Belgian patients. CONCLUSIONS: US patients used more blood products, had shorter hospital stays, and incurred greater costs than Belgian patients. Specialists as attending physicians were associated with lower utilization of ABT; this may be an administrative change that hospitals can implement to reduce utilization and costs. PMID- 17919358 TI - How do people with gastro-oesophageal reflux disease perceive their disease? Results of a multinational survey. AB - OBJECTIVE: To determine how patients with gastro-oesophageal reflux disease (GERD) perceive their condition. METHODS: Subjects with a formal diagnosis of GERD (n = 929) and those who did not have a formal diagnosis but regularly suffered from symptoms suggestive of GERD (heartburn or acid regurgitation, n = 924) were identified as part of a multinational survey. RESULTS: Symptoms had been present for a mean of 11 years in diagnosed individuals and 8 years in undiagnosed individuals. Over-the-counter (OTC) antacids were used by 78% of undiagnosed individuals. Most patients (68% of undiagnosed and 46% of diagnosed subjects) did not consider themselves to have a serious condition, while 70% of undiagnosed and 58% of diagnosed respondents believed that their condition would not have any long-term health consequences. A key perception driving the decision to seek medical care was the belief that a physician, and prescribed medication, could help. Conversely, the perception that OTC medications work effectively was the greatest barrier to visiting a physician. Diagnosed patients, who, on average, waited 2 years before seeking medical advice, indicated that prescription medication reduced the severity and frequency of their symptoms. CONCLUSIONS: There is widespread perception that GERD is a non-serious disease without long-term health consequences, and some patients find that OTC medications are a sufficiently effective treatment option. Educational initiatives should perhaps be targeted at those in greatest medical need, overcoming the barriers that dissuade many of these patients from seeking appropriate medical care. PMID- 17919359 TI - Thiazolidinedione use and post-operative atrial fibrillation: a US nested case control study. AB - BACKGROUND: Previous investigations suggested thiazolidinediones (TZDs) have the ability to suppress inflammation. Since the pathophysiology of atrial fibrillation (AF) after cardiothoracic surgery (CTS) likely involves an inflammatory mechanism, we sought to determine whether preoperative use of TZDs could further reduce the incidence of post-CTS AF in a population treated with beta-blockers and prophylactic amiodarone. METHODS: All diabetic patients over the age of 50 years, not in atrial arrhythmia prior to surgery, who underwent CTS from the Atrial Fibrillation Suppression Trials I, II and III (AFIST I, II and III) were evaluated in this nested case-control study. Patient demographics, surgical characteristics, medication utilization and the incidence of post-CTS AF (AF > 5 minutes duration) were collected as part of AFIST I, II and III. Multivariate logistic regression was used to calculate adjusted odds ratios with 95% confidence intervals (CIs). RESULTS: One hundred and eighty-four diabetic patients were enrolled in the three trials. Overall, the study population averaged 66.9 +/- 7.3 years of age, 71.7% were male, 7.1% underwent valve surgery, 4.9% had prior AF, 17.9% had heart failure and 84.2% and 41.8% received postoperative beta-blockade and prophylactic amiodarone, respectively. Forty patients (21.7%) received a preoperative TZD and 144 (78.3%) did not. In total, 66 patients (35.9%) developed post-CTS AF. Upon multivariate logistic regression, the preoperative use of TZDs was found to be associated with a 20% non statistically significant reduction in post-CTS AF (adjusted odds ratio; 0.80, 95% CI 0.32-1.99; p = 0.63). LIMITATIONS: Patients were not randomized to receive TZDs or not. We may not have had adequate power to evaluate our post-CTS AF endpoint. CONCLUSIONS: In a diabetic population treated perioperatively with beta blocker and amiodarone, adjunctive TZD use was associated with a non statistically significant reduction in a patient's odds of developing post-CTS AF. Further research is needed to determine whether TZDs, in fact, do not have anti-fibrillatory effects or whether our study was underpowered to detect a statistically significant benefit with TZDs. PMID- 17919360 TI - Suicide risk in schizophrenia: explanatory models and clinical implications, The Schematic Appraisal Model of Suicide (SAMS). AB - PURPOSE: The objective of this paper was to evaluate the Cry of Pain model of suicide behaviour as applied to psychosis and to derive theoretically driven guidelines for prevention. Suicide risk in psychotic patients is a serious but poorly understood clinical and social problem. There is a dearth of psychological models to explain suicidal behaviour and to guide effective preventative clinical interventions. Understanding suicidal behaviour in psychosis may be facilitated by utilizing models from depression. The 'Cry of Pain' model of suicide is founded on an evolutionary approach to understanding suicidal behaviour in depression. METHODS: The model was critically evaluated and relevant literature reviewed. RESULTS: Although strengths of the model were identified, lack of clarity regarding the concepts of defeat, entrapment, and lack of rescue appeared to limit its theoretical and clinical utility. We suggest a modification to this model which replaces concepts of defeat, entrapment, and lack of rescue with a four-stage appraisal process in conjunction with information-processing biases and suicidal schema. CONCLUSIONS: Methods of testing this model are suggested and guidelines for a clinical intervention (Cognitive Behavioural Suicide Prevention for Psychosis; CBSPp) are outlined. PMID- 17919361 TI - Thoracic injury: a review of 276 cases. AB - OBJECTIVE: Chest injury, one of the most important aspects of trauma, directly accounts for 25% of all trauma-related deaths and plays a major contributing role in another 25% of trauma deaths. This paper aimed to explore the spectrum and outcome of thoracic injuries seen in a multi centric study of trauma patients. METHODS: A total of 276 consecutive trauma patients in 6 general hospitals were analyzed. The feature of injury, injury severity score (ISS), clinical treatment and mortality were recorded in a prospective manner and analyzed retrospectively. Multiple logistic regression analysis was used to determine the independent predictors of mortality following the chest trauma. RESULTS: There were 246 males (89.1%) and 30 females (10.9%) ranging from 3 to 80 years with a mean age of (34+/-17) years. Road traffic accident was the main cause of injury, especially for pedestrians, followed by stab wound (89 cases, 32.1%) and falling injuries (32 cases, 11.6%), respectively. Haemothorax or pneumothorax (50.4%) and rib fracture (38.6%) were the most common types of chest injury. Extremity fracture was the most common associated injury with the rate of 37% ( 85/230), followed by head injury (25.2%) and abdominal trauma (19.6%). These injuries contributed significantly to the morbidity and mortality of trauma patients. CONCLUSIONS: According to the results, most patients with chest injury can be treated conservatively with close observation and tube thoracostomy. The presence of blunt trauma, head injury and abdominal injury independently adversely affect mortality after chest trauma. It is necessary to investigate the causes and patterns of injuries resulting from stab wound for effective prevention. PMID- 17919363 TI - Effect of radix paeoniae rubra on expression of p38 MAPK/iNOS/HO-1 in rats with lipopolysaccharide-induced acute lung injury. AB - OBJECTIVE: To investigate the effect of radix paeoniae rubra (RPR) on expression of p38 mitogen activated protein kinase (MAPK)/iNOS/HO-1 in rats with lipopolysaccharide-induced acute lung injury and explore the molecular mechanism. METHODS: Forty healthy male Wistar rats, weighing 200-250 g, aged 6-8 weeks (mean equal to 7 weeks), provided by the Experimental Center, Medical College, Wuhan University, Wuhan, China, were employed in this study. Under anesthesia with 7% chloraldurat (5 ml/kg body weight) through intraperitoneal injection, the trachea of the rat was exposed and an arterial puncture needle pricked into the trachea via cricothyroid membrane. Then they were randomly divided into five groups: 8 rats receiving 1 ml normal saline through the puncture needle (Group A), 8 receiving 1 ml lipopolysaccharide (LPS, 2.5 mg/kg, Group B), 8 receiving LPS and RPR (30 mg/kg, pumped through the femoral vein for 2 hours, Group C), 8 receiving RPR 2 hours before dripping LPS (Group D), and 8 receiving hemin (75 micromol/L through intraperitoneal injection) 18 hours before dripping LPS (Group E). After 6 hours of LPS dripping, blood samples were obtained through the carotid artery to perform blood gas analysis, then all the rats were exsanguinated to death and specimens of lung tissues were obtained. The pathomorphological changes of the lung tissues were observed. The expression of p38 MAPK/iNOS/HO-1, the neutrophil ratio, protein content in alveolar irrigating solution and malonaldehyde (MDA) content in the lung tissues were also detected. RESULTS: Compared with Group A, the expression of p38 MAPK, iNOS and HO-1 markedly increased in Groups B, C, D, and E (P < 0.01). But in Groups C, D and E the expression of p38 MAPK and iNOS were significantly lower than that of Group B, while expression of HO-1 was obviously higher than that of Group B (P < 0.05). The protein content, the ratio of neutrophils in bronchoalveolar lavage fluid (BALF), the content of MDA and the activities of serum NO in Group B were significantly higher than those of Group A (P < 0.01). There was a significant decrease in the level of arterial bicarbonate and partial pressure of oxygen in Group B (P < 0.01). Compared with Group B, these indexes of lung injury were significantly lower while the levels of arterial bicarbonate and partial pressure of oxygen increased significantly in Groups C, D and E (P < 0.05 or P < 0.01). Under light microscope, the pathological changes induced by LPS were significantly attenuated by RPR and hemin. CONCLUSIONS: The high expression of MAPK plays an important role in lipopolysaccharide-induced acute lung injury. Protective effect of RPR on lipopolysaccharide-induced acute lung injury may be related to the inhibition of the abnormal high expression of p38 MAPK/iNOS/HO-1. PMID- 17919362 TI - Variations of p38 MAPK and sICAM-1 with therapeutic effect of different resuscitation fluids on severe traumatic patients. AB - OBJECTIVE: To investigate the dynamic variation and action mechanism of sICAM-1 and p38 mitogen-activated protein kinases (MAPK) signal transduction in human severe trauma and resuscitation, as well as the effect of lactated Ringer's solution(LR), 7.5% sodium chloride solution(HS) and 20% albumin injection(ALB) on the incidence and mortality of multiple organ dysfunction syndrome (MODS). METHODS: Seventy-two severe trauma patients (ISS score 16-43) were divided into ISS < or = 25 and ISS > 25 groups (each group was subdivided into LR, HS and ALB groups). ELISA was used to measure the concentration of sICAM-1. Western blot was used to measure the expression of p38 MAPK. RESULTS: Compared with LR group, the transfusion volume needed for maintaining systolic blood pressure > or = 90 mm Hg was significantly decreased in HS and ALB groups (P < 0.05). Compared with the control group, the concentration of blood sICAM-1 and the expression of p38 MAPK was elevated from 4 to 48 hours after trauma in all experimental groups (P < 0.05 0.01). At 4, 12, and 24 hours, there was significant correlation between the expression of p38 MAPK and sICAM-1 (P < 0.01). Compared with LR group, sICAM-1 and p38 MAPK in HS and ALB groups were decreased (P < 0.05). sICAM-1 and p38 MAPK were significantly higher in the group of ISS > 25 than that of ISS < or = 25 (P < 0.05). MODS incidence and mortality were significantly higher in the group of ISS > 25 than that of ISS < or = 25 (P < 0.05). MODS incidence and mortality were lower in HS and ALB groups than LR group (P < 0.05). CONCLUSIONS: The up regulation of polymorphonuclear neutrophil-endotheliocytes (PMN-EC) adhesion may be due to the increased sICAM-1 expression during severe trauma. The up regulation of sICAM-1 expression is correlated with the activation of p38 MAPK. During severe trauma, the levels of sICAM-1 and p38 MAPK, as well as the incidence and mortality of MODS are lower when HS and ALB are used than single lactated LR solution is used. PMID- 17919364 TI - Analysis of high risk factors related to acute respiratory distress syndrome following severe thoracoabdominal injuries. AB - OBJECTIVE: To investigate the high risk factors related to acute respiratory distress syndrome(ARDS) following serious thoracoabdominal injuries. METHODS: The clinical data of 282 patients with serious thoracoabdominal injuries were retrospectively studied. Univariate and Cox multivariate regression analysis were used to determine the risk factors related to ARDS following serious thoracoabdominal injuries. RESULTS: The incidence of ARDS was 31.9% (90/282) in patients with serious thoracoabdominal injuries. The mortality caused by ARDS was 37.8% (34/90). The univariate analysis and multivariate analysis demonstrated that the clinical conditions such as elder age, shock, dyspnea, abnormal arterial blood gas, hemopneumothorax, pulmonary contusion, flail chest, coexisting pulmonary diseases, multiple abdominal injury and high ISS score were the independent high risk factors related to ARDS. CONCLUSION: There are many high risk factors related to ARDS following severe thoracoabdominal injuries, which should be detected early and treated timely to decrease the incidence and mortality of A RDS. PMID- 17919365 TI - Damage control surgery for severe thoracic and abdominal injuries. AB - OBJECTIVE: To investigate the application of damage control surgery in treatment of patients with severe thoracic and abdominal injuries. METHODS: A retrospective study was done on 37 patients with severe thoracic and abdominal injuries who underwent damage control surgery from January 2000 to October 2006 in our department. There were 8 cases of polytrauma (with thoracic injury most commonly seen), 21 of polytrauma (with abdominal injury most commonly seen) and 8 of single abdominal trauma. Main organ damage included smashed hepatic injuries in 17 cases, posterior hepatic veins injuries in 8,pancreaticoduodenal injuries in 7, epidural or subdural hemorrhage in 4, contusion and laceration of brain in 5, severe lung and bronchus injuries in 4, pelvis and one smashed lower limb wound in 3 and pelvic fractures and retroperitoneal hemorrhage in 6. Injury severity score (ISS) was 28-45 scores (38.4 scores on average), abbreviated injury scale (AIS) > or = 4.13. The patients underwent arteriography and arterial embolization including arteria hepatica embolization in 4 patients, arteria renalis embolization in 2 and pelvic arteria retroperitoneal embolization in 7. Once abbreviated operation finished, the patients were sent to ICU for resuscitation. Twenty-four cases underwent definitive operation within 48 hours after initial operation, 5 underwent definitive operation within 72 hours after initial operation, 2 cases underwent definitive operation postponed to 96 hours after initial operation for secondary operation to control bleeding because of abdominal cavity hemorrhea. Two cases underwent urgent laparotomy and decompression because of abdominal compartment syndrome and 2 cases underwent secondary operation because of intestinal fistulae (1 case of small intestinal fistula and 1 colon fistula) and gangrene of gallbladder. RESULTS: A total of 28 patients survived, with a survival rate of 75.68%, and 9 died (4 died within 24 hours and 5 died 3-9 days after injury). The trauma deaths at the early stage were caused by severe primary injuries resulting in failure of respiration and circulation, while mortality at the later stage was caused by multiple organ failure. CONCLUSIONS: Damage control surgery is important for the treatment against severe thoracic and abdominal injuries. It is suggested that the surgeon should select the reasonable auxiliary examination before operation, and take the proper time to perform damage control and definitive surgery. PMID- 17919366 TI - Treatment of abdominal injuries: a report of 522 cases. AB - OBJECTIVE: To make a summary of the experiences in the treatment of abdominal injuries. METHODS: A retrospective study was done on 522 cases of abdominal injuries in our department from January 1986 to December 2004. RESULTS: Of all,382 cases were treated by surgery and 140 by conservative method. Among the surgically treated cases, 347 patients (90.8%) recovered, 35(9.2%) died and 21 had postoperative complications (5.6%). For patients undergoing conservative treatment, 139(99.3%) recovered but one (0.7%) died. CONCLUSIONS: The severity of abdominal injury and delayed treatment are two key factors leading to death. Surgical procedure is still the main method against alternative abdominal injuries. It is necessary to strictly control the indications in conservative treatment. PMID- 17919367 TI - Construction of eukaryotic expression plasmid human transforming growth factor beta3 and its transfection into precartilaginous stem cells. AB - OBJECTIVE: To obtain seed cells for cartilage repair through constructing recombinant human transforming growth factor beta3 vector (hTGF-beta3) and transfecting it into rat's precartilaginous stem cells (PSCs). METHODS: Gene engineering technique was introduced to construct eukaryotic expression plasmid pcDNA3.1 (+)-hTGF-beta3. PSCs of rats were isolated and purified with method of immunomagnetic microbeads. Then PSCs were cotransfected with plasmid hTGF-beta3 and pcDNA3.1 (+)-enhanced green fluorescence protein (EGFP) by liner polyethyleneimine (PEI). And 48 hours later the transient expression of EGFP was observed under a fluorescence microscope, and the expression of hTGF-beta3 was detected with reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA). RESULTS: The sequences of the recombinants were consistent with that from Genebank. Cotransfection of EGFP provided fast visual confirmation of successful transduction. The hTGF-beta3 mRNA and protein expression could be detected by RT-PCR and ELISA. CONCLUSIONS: The recombinant plasmid is correctly constructed and successfully transfected into rat's PSCs, which is an important step to treat epiphyseal injury or other osteo-cartilage diseases with transgenic therapy. PMID- 17919368 TI - Brachial plexus palsy caused by halo traction before posterior correction in patients with severe scoliosis. AB - OBJECTIVE: To explore the clinical features and treatment results of brachial plexus palsy caused by halo traction before posterior correction in patients with severe scoliosis. METHODS: A total of 300 cases of severe scoliosis received halo traction before posterior correction in our department from July 1997 to November 2004. Among them, 7 cases were complicated with brachial plexus palsy. The average Cobb angle was 110 degree (range, 90 degree-135 degree). Diagnoses were made as idiopathic scoliosis in 1 case, congenital scoliosis in 3 cases, and neuromuscular scoliosis in 3 cases. Additionally, diastematomyelia and tethered cord syndrome were found in 3 cases and thoracolumbar kyphosis in 2 cases. Weight of traction was immediately reduced when the patient developed any abnormal neurological symptoms in the upper extremity, and rehabilitation training was undertaken. Simultaneously, neurotrophic pharmacotherapy was applied, and the neurological function restoration of the upper limbs and the recovery time were documented. RESULTS: Traction was used for an average of 3.5 weeks (range, 2-6 weeks) before spinal fusion for these 7 patients. The average traction weight was 8 kg, which was 19% on average (range, 13%-26%) of the average body weight (40.2 kg). These 7 patients had long and thin body configuration with a mean height of 175 cm. The duration between symptoms of brachial plexus paralysis and the diagnosis was 1-3 hours. All of these 7 patients presented various degrees of numbness in the ulnar side of the hand and forearm. Median nerve paresis was found in 3 cases and ulnar nerve paresis in 4 cases. Complete recovery of the neurological function had been achieved by the end of three months. CONCLUSIONS: The clinical features of brachial plexus palsy caused by halo traction include median nerve paresis, ulnar nerve paralysis, and numbness in the ulnar side of the hand and forearm, which may be due to the injury of the inferior part of the brachial plexus, i.e., damage of C8 and T1 nerve roots. Complete recovery of neurological function can be expected when the patient is kept under careful observation for recognizing this complication as soon as possible, then immediately reducing or removing the traction weight, and adopting rehabilitation training and neurotrophic pharmaceutical treatment. PMID- 17919369 TI - Applied anatomy of the lower cervical pedicle screw insertion. AB - OBJECTIVE: To ascertain an accurate approach to inserting the pedicle screw into C3-C7 segments of the cervical vertebra. METHODS: Anatomic morphology of lateral mass and pedicle, and their anatomic relationship with the adjacent tissue were observed on C3-C7 segments of 25 adult embalmed cadavers (50 sides). RESULTS: 1) The inferior edge of the base of the posterior tubercle of the transverse process and the inferior edge of the pedicle were connected with each other on 25 adult embalmed cadavers (50 sides). The transverse section which passed through the median point between the superior edge and the inferior edge of the base of the posterior tubercle of the transverse process, and the transverse section which passed through the central axis between the superior edge and the inferior edge of the pedicle, were in the same horizontal plane. The superior and inferior position of placing the pedicle screw was determined by this transverse section, which passed through the median point between the superior and the inferior edge of the base of the posterior tubercle of the transverse process. 2) There was a directed internal-downwards "triangular sulcule" between the base of the posterior tubercle of the transverse process and the anterolateral edge of the inferior articular process. The anterior wall of the triangular sulcule was the base of the posterior tubercle of the transverse process, the posterior wall was the anterolateral edge of the inferior articular process, and the bottom of the sulcule was connected with the interior edge of the pedicle. The vertical length between the top of triangle and the planes of inferior edge of the pedicle was (2.78+/-1.71) mm. The inferior edge of the cervical pedicle could be detected using a blunt probe along the "triangular sulcule" between the base of the posterior tubercle of the transverse process and the anterolateral edge of the inferior articular process in surgical operation. 3) The lateral fovea of the articular process was observed on all lateral masses (50 sides). The internal and external position of the entrance point could depend on anatomic landmarks: the lateral edge of the lateral fovea of the articular process. The horizontal length between the lateral fovea of the articular process and the entrance point was (3.14+/-1.45) mm. 4) The diameter of pedicle screw, about (2.78+/-1.71) mm, was the transverse diameter of the cancellous bone of the greatest narrow part of the cervical pedicle. CONCLUSIONS: The median point between the superior edge and the inferior edge of the base of the posterior tubercle of the transverse process, the lateral fovea of the articular process, and the triangular sulcule between the base of the posterior tubercle of the transverse process and the anterolateral edge of inferior articular process, are easy to be exposed and identified in surgical operation. The pedicle screw can be precisely inserted through this method. PMID- 17919370 TI - Effect of fluvastatin on vascular endothelial growth factor in rats with osteoporosis in process of fracture healing. AB - OBJECTIVE: To explore the effect of fluvastatin on vascular endothelial growth factor (VEGF) in rats with osteoporosis in the process of fracture healing. METHODS: Fractures at the intermediate piece of the femur were made on 72 Sprague Dawley (SD) rats (weighing initially 290-340 g and aged 6 months) with osteoporosis after ovariectomy for three months, then these rats were divided randomly into the medication administration group (the experimental group) and the control group, 36 rats each. In the experimental group, the rats received fluvastatin lavage (10 mg/kg per day) since the next day of operation lasting for 6 weeks, and the rats in the control group received placebo. Then the expression of VEGF and VEGF mRNA in bony callus of the two groups was measured respectively with immunohistochemistry and in situ hybridization on days of 3rd, 7th, 14th, 21st, 28th, and 42nd, and image analysis was made with real-color image analysis machine. RESULTS: No difference was found in the cellular localization of VEGF and VEGF mRNA gene expression between the experimental group and the control group in process of fracture healing and their expression modes were almost similar. On the 14th day postoperatively, the positive extent of positive cells in the experimental group was higher than that of the control group (P < 0.05). CONCLUSION: Fluvastatin can promote the VEGF level in rats with osteoporosis in process of fracture healing. PMID- 17919371 TI - Microendoscopic discectomy, a less traumatic procedure for lumbar disk herniation. AB - OBJECTIVE: To investigate the change of serum levels of interleukin-6 (IL-6), C reactive protein (CRP) and creatine kinase (CK) in patients undergoing microendoscopic discectomy (MED) and open discectomy. METHODS: Forty-four patients with single level lumbar disk herniation were treated, either by MED (Group A, n equal to 22) or open discectomy (Group B, n equal to 22). Peripheral venous blood samples were taken before surgery and at 24 and 48 hours postoperatively. The operating time, intraoperative blood loss, postoperative hospital stay were recorded. The pain severity of incision was evaluated by visual analog scale after operation and the clinical outcome was evaluated by Oswestry disability index. Statistical comparison was performed by the analysis of variance and Student's t test. RESULTS: The data showed that patients in Group A had a less intraoperative blood loss (P < 0.05), shorter operating length (P < 0.05), shorter postoperative hospital stay (P < 0.05) and less postoperative pain of incision than those in Group B. Serum levels of IL-6 (mean, 31.60 ng/L +/- 9.88 ng/L vs 39.16 ng/L +/- 11.14 ng/L, P < 0.05) and CK (mean, 167.91 U/L +/- 51.85 U/L vs 401.55 U/L +/- 108.86 U/L, P < 0.05) all get to the peak at 24 hours after operation and Group A with the response statistically less than Group B. Serum level of CRP peaked at 24 hours in Group A (mean, 12.68 mg/L +/- 7.10 mg/L vs 20.82 mg/L +/- 8.79 mg/L, P less than 0.05)and peaked at 48 hours after surgery in Group B (mean, 10.77 mg/L +/- 5.25 mg/L vs 29.95 mg/L +/- 14.85 mg/L, P < 0.05). The clinical outcomes of both groups were the same at 6 months after surgery. CONCLUSIONS: Both MED and open discectomy have made good clinical outcomes, however, the less change of IL-6, CRP and CK after operation proves that MED procedure is less traumatic to patients than open discectomy. PMID- 17919372 TI - Plastic materials currently used in Mongolia. AB - The first skin-flap procedures were performed in Mongolia by a team of Chinese doctors (Chenod, Chen) who treated children with burn injuries in the 1950s. The field of plastic surgery was further developed through assistance from the former Soviet Union and is now widely practiced by a number of surgeons around the country. In recent years, the fast evolving field of plastic surgery in Mongolia has created a need for clear and consistent system for the classification of various plastic materials. A team of Mongolian surgeons at the National Hospital for Traumatology, Orthopedics, Rehabilitation and Teaching Research have completed a research programme aimed at adopting a leading classification system that can facilitate effective communication between plastic surgeons. This requires the chosen system to be in line with modern trends in plastic surgery and the established international norms. As a result of extensive research and analysis, they have developed a customized version of Vasiliev's classification that focuses on formulating general principles of the description of plastic materials based on their functional characteristics. The main reason for this selection is its similarity with existing Mongolian standards as well as its principles that contain important implications for surgery. PMID- 17919373 TI - Foreign metallic pellet in the heart. AB - Foreign bodies in the heart are a rare but serious form of cardiac injury. The objects usually are sharp pointed. Such as acupuncture needles, sewing needles, coat hangers, fragments of Kirschner wires, pins, etc.(1) We report a patient with a metallic pellet lodged in the heart, which was accurately diagnosed and successfully removed. PMID- 17919374 TI - [Impact of hypocaloric and hypo-nitrogen parenteral nutrition on clinical outcome in postoperative patients: a multi-center randomized controlled trial of 120 cases]. AB - OBJECTIVE: To evaluate the impact of hypocaloric and hypo-nitrogen parenteral nutrition (PN) on infective complication rate, postoperative hospital stay and treatment cost in postoperative period. METHODS: 120 patients with gastrointestinal tumors with the Nutrition Risk Screening (NRS) score of 3 or 4 undergoing radical gastrectomy in 5 hospitals were randomly assigned into 2 equal groups: control group, receiving PN with 30 (28 - 32) kcal x kg(-1) x d(-1) and nitrogen 0.20 (0.19 - 0.21) g x kg(-1) x d(-1) in regular "3 liter bag", and study group receiving calorie of 18 (range 16 - 20) kcal x kg(-1) x d(-1) and nitrogen of 0.10 (0.09 - 0.11) g x kg(-1) x d(-1) with triple chamber bag. PN support was infused continuously for at least six postoperative days through peripheral vein or peripherally inserted central catheter. The differences between these two groups in blood glucose level, infectious complication, phlebitis, systemic inflammatory response syndrome (SIRS), and duration of hospital stay after operation, and treatment cost. All data were evaluated by both intention to treat (ITT) analysis and per protocol (PP) analysis. RESULTS: There were no significant differences in the clinical baseline and operative types between the two groups. ITT analysis showed that the occurrence of hyperglycemia in postoperative period in the control group was 43.3%, significantly much higher than that in the study group (6.6%, P = 0.000). The infectious complication rate of the study group was 3.3%, significantly lower than that of the control group (16.6%, P = 0.0149), the phlebitis rate of the study group was 0.0%, significantly lower than that of the control group (18.3%, P = 0.0005). The SIRS rate of the study group was 25.0%, significantly lower than that of the control group (45.0%, P = 0.0216). PP analysis showed that the postoperative duration of hospital stay of the control group was 14.1 days +/- 5.8 days, significantly longer than that of the study group (12.4 days +/- 4.0 days, P = 0.047), the total PN cost of the study group was 3411.6 +/- 181.1 Yuan RMB, significantly higher than that of the control group (2945 +/- 162 Yuan RMB, P = 0.000); but the total post-operative cost of treatment of the control group was 13156 +/- 3282 Yuan RMB, significantly higher than that of the study group (11 642 +/- 3019 Yuan RMB, P = 0.010); and the time for compounding of the study group was 5.0 min +/- 1.7 min, significantly shorter than that of the control group (15.4 min +/- 3.7 min, P = 0.000). CONCLUSION: Hypocaloric and hypo nitrogen PN in postoperative days 1 - 6 in patients with scores 3 or 4 decreases the rates of hyperglycemia, infectious complications, phlebitis, and SIRS, shortens the postoperative hospital stay, and lowers the cost of treatment comparing with conventional PN. The use of triple chamber bag shortens the compounding time of PN. PMID- 17919375 TI - [Association between metabolic enzyme genotype of azathioprine and drug tolerance in patients with rheumatic diseases]. AB - OBJECTIVE: To discuss the relationship between the genotype of thiopurine methyltransferase (TPMT) and azathioprine tolerance in the patients with rheumatic diseases. METHODS: Four common mutation alleles of TPMT in 200 patients with rheumatic diseases [TPMT*2 (G238C), TPMT*3A (A719G/G460A), TPMT*3B (G460A), TPMT*3C (A719G)] were detected by allele specific polymerase chain reaction (AS PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR PFLP). 194 patients who had used azathioprine finished the 3 months' follow-up. RESULTS: In the 200 patients with rheumatic diseases, 4 cases of heterozygote of TPMT*3C were detected, but no mutation of TPMT*2, TPMT*3B or TPMT*3A was found. The genotypic frequency of wild-type homozygote was 98%, and that of heterozygote (TPMT*1/TPMT*3C) was 2%. Mutation allele frequency in these patients was 1%. Average of TPMT activity of the 4 cases of heterozygote was (2.4 +/- 1.2) U/ml red blood cells, significantly lower than that of the 196 cases of homozygote which was (12.2 +/- 6.8) U/ml RBC. In the 194 patients who had used azathioprine, bone marrow suppression occurred in 18 patients, 2 of which suffered severe crisis of hematopoietic system, and 6 of which were complicated with impaired liver function. In the 4 patients with heterozygote, 3 had used azathioprine, and bone marrow suppression occurred within 1 month of using the drug, including 2 cases of severe crisis of hematopoietic system. CONCLUSION: Patients with mutation alleles of TPMT are intolerant to azathioprine, and likely to have severe crisis of hematopoietic system. To detect the TPMT genotype before using azathioprine is significant to improve the therapeutic safety. PMID- 17919376 TI - [The expression of MRP, GST-pi, Topo IIalpha and COX-2 in epithelial ovarian cancer and its relationship to drug resistance and prognosis]. AB - OBJECTIVE: To study the expression of MRP, GST-pi, Topo IIalpha and COX-2 in epithelial ovarian cancer (EOC) and its relationship to drug resistance and prognosis. METHODS: Immunohistochemical method was used to study the expression of MRP, GST-pi, Topo IIalpha and COX-2 in 60 cases of epithelial ovarian cancer who were treated in PUMC Hospital from 2000 to 2004. The mean age of all patients was 51.2 years old. The histological types were 35 cases of serous adenocarcinoma, 2 cases of mucinous adenocarcinoma, 11 cases of clear cell carcinoma, 4 cases of transitional cell carcinoma and 8 cases of endometrioid carcinoma. Statistical analysis was done to determine its relationships to drug resistance and prognosis. RESULTS: The positive expression rate of MRP, GST-pi, Topo IIalpha and COX-2 in EOC was 26.7%, 93.3%, 61.7% and 78.3% respectively. The positive expression rate of MRP was significantly higher in clear cell carcinoma than that in non-clear cell carcinoma (54.5% vs 20.4%, P < 0.05); The expression of Topo IIalpha in advanced stage of EOC was significantly lower than that in early stage cases (56.6% vs 78.6%, P < 0.05). Positive expression of MRP, negative expression of Topo IIalpha, strong positive expression of GST-pi and COX 2 were seen as four risk factors for recurrence, drug-resistance and poor prognosis. More cases of recurrent group and drug-resistant group had 2 or more risk factors. The three-year survival rate of patients with 2 or more risk factors and patients with less than 2 risk factor are 69.9% and 91.7% respectively (P > 0.05). CONCLUSIONS: There are moderate or high expression of MRP, GST-pi, Topo IIalpha and COX-2 in epithelial ovarian cancer. Having 2 or more risk factors may be seen as a vital indicator to predict drug-resistance and recurrence in patients of epithelial ovarian cancer. PMID- 17919377 TI - [Intradiscal electrothermal treatment combined with epidural steroid injection for discogenic lumbago-leg pain]. AB - OBJECTIVE: To assess the effectiveness of intradiscal electrothermal treatment (IDET) combined with epidural steroid injection for discogenic lumbago-leg pain. METHODS: Fifty-nine patients with discogenic lower back pain and radical pain underwent IDET and CT guided spinal cord epidural steroid injection (Limethason). Thirty-eight patients with discogenic lower back pain and radical pain underwent IDET only. The clinical efficacy was observed within 1 week, 1 week-1 month, 1 - 3 months, and 3 - 6 months after the treatment. RESULTS: The efficacy rates within 1 week, 1 week-1 month, 1 - 3 months, and 3 - 6 months after the treatment of the IDET plus epidural steroid injection group were 71%, 55%, 81%, and 89% respectively, and those of the IDET only group were 55%, 34%, 76% and 87% respectively. The efficacy rate 1 week-1 month postoperatively of the IDET plus epidural steroid injection group was significantly higher than that of the IDET only group (chi(2) = 4.375, P < 0.05), and the efficacy rates at other time points were not significantly different between these 2 groups. CONCLUSION: IDET combined with epidural steroid injection relieves post-IDET edema and induces pain more effectively. PMID- 17919378 TI - [Effects of cattle encephalon glycoside and ignotin injection on serum level of S100B protein and neuron specific enolase in patients undergoing cardiac valve replacement with cardiopulmonary bypass]. AB - OBJECTIVE: To investigate the effects of cattle encephalon glycoside and ignotin injection on serum level of S100B protein and neuron specific enolase (NSE) during cardiac valve replacement with cardiopulmonary bypass (CPB). METHODS: Forty patients undergoing cardiac valve replacement with CPB were randomly divided into two equal groups: cattle encephalon glycoside and ignotin injection group (Group A, injected intravenously with cattle encephalon glycoside and ignotin 20 ml after induction of anesthesia, and control group (Group B, injected intravenously with normal saline instead. Blood samples were taken from the jugular bulb after induction of anesthesia (T1), immediately, and 2 h, 6 h, and 24 h after the discontinuation of CPB (T2, T3, T4, and T5) for determination of serum levels of S100B protein and NSE. RESULTS: The serum levels of S100B protein and NSE of both groups were significantly increased after CPB, and then gradually decreased. At the time point T5, the serum levels of S100B protein of both groups decreased to near the baseline levels (T1 levels), but the serum levels of NSE remained high in both groups. At the time points T2, T3, T4 and T5, the serum levels of S100B protein were significantly higher in Group B than in Group A (all P < 0.05). At the time points T2, T3, and T4, the serum levels of NSE was significantly higher in Group B than in Group A (all P < 0.05). CONCLUSION: Cattle encephalon glycoside and ignotin injection decreases the serum levels of S100B protein and NSE and has cerebral protective effect during cardiac valve replacement with CPB to a certain degree. PMID- 17919379 TI - [Effects of pleural drainage using central venous catheter and simple aspiration in treatment of spontaneous pneumothorax: a clinical comparative study]. AB - OBJECTIVE: To compare the curative effects and expenses of pleural drainage by using central venous catheter and of simple aspiration by thoracentesis in treatment of spontaneous pneumothorax. METHODS: 46 patients with spontaneous pneumothorax were randomly divided into 2 equal groups: Group A, treated with pleural drainage by using central venous catheter, and Group B, treated with simple aspiration by thoracentesis. The curative rate and average residual volume after treatment were used to evaluate the curative effects. The average hospitalization days, duration of treatment, and expenses for medical materials were used to evaluate as the economic index. RESULTS: (1) The effective rates were 100% and 91.30% in Groups A and B respectively. (2) The average residual volume of pneumothorax at the time of discharge from hospital of Group A was 4.17%, significantly lower than that of Group B (18.62%, P < 0.05). (3) The average residual volume of pneumothorax 1 week after discharge of Group A was 0.53%, significantly lower than that of Group B (7.59%, P < 0.01). (4) The expense for medical materials of Group A was 264.79 Yuan RMB, a little bit more than that of Group B (233.62 Yuan RMB). (5) The hospitalization time of Group A was 5.32 days, significantly shorter than that of Group B (7.68 days, P < 0.05). The duration of treatment with antibiotics of Group A was 3.46 days, significantly shorter than that of Group B (5.59 days, P < 0.05). The time of oxygen inhalation of Group A was 86.4 hours, significantly shorter than that of Group B (133.1 hours, P < 0.01). CONCLUSION: With higher curative effect and lower expenses, pleural drainage by using central venous catheter is superior to simple aspiration in treatment of spontaneous pneumothorax, and can be the first choice. PMID- 17919380 TI - [Clinical observation of high thoracic epidural anesthesia therapy for patients with congestive heart failure secondary to ischemic cardiomyopathy]. AB - OBJECTIVE: To investigate the effects of high thoracic epidural anesthesia (HTEA) on chronic heart failure (CHF) and its safety. METHODS: 117 patients with ischemic cardiomyopathy (ICM) of NYHA class II - IV and with the LVEF < or = 45%, 90 males and 27 females, aged 45 - 81, were Randomly divided into 2 groups: control group (n = 54, treated with the routine medication) and HTEA group (n = 63, administrated epidurally with 0.5% lidocaine 3 - 5 ml every two hours from 9 a.m. to 11 p.m. everyday via an epidural catheter for one month in addition to the routine treatment). Echocardiography was performed before and after the treatment to examine the cardiac structure and left ventricular systolic function. The plasma brain natriuretic peptide (BNP) was examined before and after the treatment. RESULTS: The clinical symptoms and signs of the HTEA group were improved markedly. The total effective rate was 95.2% in the HTEA group, significantly higher than in the control group (85.2%, P < 0.05). Echocardiographic parameters showed that the diameters of ventricles and atrium decreased by 3 - 5 mm in the HTEA group. The left ventricular end diastolic volume (LVEDV) of the HTEA group reduced from (337 +/- 132) ml before treatment to 274 ml +/- 109 ml after treatment, and the left ventricular end systolic volume (LVESV) of the HTEA group reduced from (215 +/- 99) ml before treatment to (147 +/- 73) ml after treatment (both P < or = 0.001). The regurgitation area of mitral valve of the HTEA group reduced from (6.6 +/- 3.7) cm(2) before treatment to (4.3 +/- 2.7) cm(2) after treatment, and the regurgitation area of tricuspid valve of the HTEA group reduced from (5.4 +/- 3.2) cm(2) before treatment to (3.3 +/- 2.3) cm(2) after treatment (both P < 0.01). No changes were observed in the E/A ratio in both groups. The BNP level fell from (678 +/- 521) ng/L to (237 +/- 225) ng/L in the HTEA group (P < or = 0.05). In the HTEA group, one patient died because of deterioration of cardiac function. HTEA was well tolerated and caused lower incidence of adverse events. CONCLUSIONS: HTEA obviously improves the heart function of the patients with CHS secondary to ICM. Consequently, HTEA is effective and safe in CHF patients with favorable tolerance. PMID- 17919381 TI - [The relationship between resistin and the peripheral neuropathy in type 2 diabetes]. AB - OBJECTIVE: To investigate the relationship between the serum resistin concentration and type 2 DM with peripheral neuropathy. METHODS: Ninety-six patients of type 2 diabetes mellitus (DM) among which 52 were with peripheral neuropathy (DM + PN) and 34 healthy persons undergoing routine physical examination as controls were studied. Body weight, waist circumference, blood pressure, body mass index (BMI), fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), and beta-cell action measured by the homeostasis model assessment (HOMA-beta), and blood lipids were measured. ELISA was used to detect the serum resistin, Endothelins (ET), and nitrogen oxide (NO). RESULTS: The serum resistin level of the DM + PN patients was (26 +/- 9) ng/ml, significantly higher than that of the DM patients [(20 +/- 6) ng/ml, P < 0.05]. Th serum resistin level was positively correlated with FINS, Homa-IR, systolic blood pressure, diastolic blood pressure, and ET (all P < 0.05) and significantly negatively correlated with HOMA-beta and NO (both P < 0.05). CONCLUSION: Resistin may play a role in the pathogenesis of type 2 DM and diabetic peripheral neuropathy. PMID- 17919382 TI - [Experience in lung resection synchronous with off-pump coronary artery bypass grafting in patients with lung tumor and coronary disease]. AB - OBJECTIVE: To summarize the experience in lung resection synchronous with off pump coronary artery bypass grafting (OPCABG) in patients with lung tumor and coronary disease. METHODS: The clinical data of ten patients with coronary artery disease and lung tumor, 8 with peripheral malignant tumors and 2 with central benign tumors, all males, aged 56.2 (41 - 782), who underwent lung resection and OPCABG synchronously, were analyzed. RESULTS: There was no hospital death in this series. All patients regained their ideal cardiorespiratory function postoperatively. CONCLUSION: With experienced management, it is feasible and safe to perform lung resection and OPCABG synchronously for strictly selected patients. PMID- 17919385 TI - [Clinical significance of atypical squamous cells and low grade squamous intraepithelial lesions in cervical smear]. AB - OBJECTIVE: To investigate the clinical significance of atypical squamous cells (ASCUS) and low-grade squamous intraepithelial lesions (LSIL). METHODS: A total of 800 patients cytologically diagnosed with ASCUS and LSIL were referred for colposcopy. The histopathology diagnosis undergoing colposcopic biopsy, endocervical curettage (ECC), cervical loop electrosurgical excision procedure (LEEP) were analyzed. Using pathology as gold criterion, the sensitivity, specificity and positive predictive value of colposcopy to detect cervical lesions and cervical cancer were measured. The follow-up results were recorded. RESULTS: (1) Among the 405 patients with ASCUS, the percentage with chronic cervicitis was 57.04%, LSIL was 34.81%, HSIL was 3.95%, otherwise 2 cases (0.49%) of microinvasive cervical cancer and 15 cases (3.70%) of vulvar intraepithelial neoplasia (VIN) were found respectively. Among the 395 patients with LSIL, the percentage with chronic cervicitis was 34.18%, LSIL was 50.89%, HSIL was 10.38%, 2 cases (0.51%) of microinvasive cervical cancer, 1 case (0.25%) of vulva squamous cancer and 15 cases (3.80%) of VIN was identified respectively. (2) The impression undergoing colposcopy was consistent with the histologically diagnosis in 637 of 800 cases (79.63%). The sensitivity, specificity and positive predictive value was 96.17%, 58.41% and 73.34% respectively. (3) All patients with HSIL or above regressed to normal after 1 year of follow-up. 501 of 738 patients with chronic cervicitis, LSIL and VIN were followed up more than 1 year. CONCLUSION: ASCUS and LSIL does not represent a single biologic entity; it subsumes changes that are unrelated to neoplasia as well as findings that suggest the possible presence of underlying Cervical intraepithelial neoplasia (CIN) and rarely carcinoma. Thorough evaluation using colposcopy will detect early not only the histological cervical HSIL and cervical cancer, but also VIN and vulvar squamous cancer. Colposcopy is a viable option in management patients with ASCUS/LSIL. PMID- 17919384 TI - [Expression of survivin gene in thyroid carcinoma and its significance]. AB - OBJECTIVE: To study the expression of survivin gene in thyroid carcinoma and its significance. METHODS: Samples of thyroid carcinoma were collected from 98 patients during operation. Western blotting and RT-PCR were used to detect mRNA and protein expression of survivin in the carcinoma tissue and the tissue around the carcinoma. TUNEL method was used to detect the apoptotic index (AI). RESULTS: Western blotting and RT-PCR methods showed similar results. The positive expression rates of survivin protein and mRNA in the thyroid carcinoma were both 68.37%; however, survivin was not expressed in all of the tissues around the thyroid carcinoma at both the mRNA and protein levels. The expression rate of survivin was closely related to the pathological typing, lymph node metastasis, and hematogenous metastasis. Apoptotic cells were found in all thyroid carcinoma. The mean AI of the surviving positive group was 0.48 +/- 0.38, significantly lower than that of the survivin negative group (2.98 +/- 0.12, P = 0.0082). CONCLUSION: Survivin gene is expressed in most of the thyroid carcinoma. The positive expression rate of survivin is significantly linked to cell proliferating activity, lymph node metastasis, and hematogenous metastasis. Survivin gene will be a new target of gene therapy in thyroid carcinoma. PMID- 17919386 TI - [The role of external signal regulated kinase and transforming growth factor beta(1) in asthma airway remodeling and regulation of glucocorticoids]. AB - OBJECTIVE: To study role of external signal regulated kinase (ERK) and transforming growth factor beta(1) (TGF-beta1) in asthma airway remodeling and to explore the regulation of glucocorticoids on ERK, TGF-beta1, and airway remodeling. METHODS: Thirty SD rats were randomly divided into 3 equal groups: control group; asthma group, undergoing intra-peritoneal injection of ovalbumin (OVA) on days 1 and 8 and inhalation of OVA every other day for 8 weeks since day 15 to establish chronic asthma models; dexamethasone (DM) intervention group, undergoing intra-peritoneal injection of DM 30 min before every inhalation instigation; and control group, receiving normal saline instead of DM. 1 - 2 hours after the last instigation the left lungs were taken out. The total bronchial wall thickness (Wat) and smooth muscle thickness (Wam) were measured by image analysis system. Phosphorylated ERK (P-ERK) was detected by immunohistochemistry. 1 - 2 hours after the last instigation blood samples were collected from the femoral artery. The concentration of transforming growth factor (TGF)-beta1 in the serum was measured by sandwich ELISA. Rat airway epithelial cells were cultured, stimulated with platelet-derived growth factor-BB (PDGF-BB, 1, 10, 25, or 50 microg/L), U0126 (specific inhibitor of phosphorylation of ERK), or budesonide (BUD). Western blotting was used to detect the P-ERK level. The level of TGF-beta1 in the cell culture supernatant was detected by sandwich ELISA. RESULTS: The Wat and Wam of the asthma group was significantly higher than those of the control group (both P < 0.01), and the Wat and Wam of the DM group were both significantly lower than those of the asthma group (both P < 0.01). The mean optical density of P-ERK and concentration of TGF beta1 in the serum of the asthma group were 31.1 +/- 2.2 and 28.1 +/- 7.4 microg/L respectively, both significantly higher than those of the control group (12.8 +/- 2.4 and 13.6 +/- 2.7 microg/L respectively, both P < 0.01), and the mean optical density of P-ERK and concentration of TGF-beta1 in the serum of the DM group were 18.7 +/- 3.1 and 15.0 +/- 3.2 microg/L respectively, both significantly lower than those asthma group (both P < 0.01). In the PDGF-BB (25 microg/L) stimulated cells marked phosphorylation of ERK occurred 15 min later, the level of P-ERK remained high up to 8 hour later, and the maximal activation occurred at the period of 2 h - 4 h later, 6.5 +/- 0.4 times that of the control value (P < 0.01). The phosphorylation levels of ERK depended on the concentration of PDGF-BB and the maximal level phosphorylation was detected with the concentration of PDGF-BB of 50 microg/L, which was 4.1 +/- 0.3 times that of the control value (P < 0.01). U0126 and BUD inhibited the phosphorylation of ERK in the cells stimulated by PDGF-BB of the concentration of 25 microg/L. there was no difference in the level of TGF-beta1 in the cell culture supernatant among different groups. CONCLUSION: Phosphorylation of ERK and TGF-beta1 have an important role in asthma airway remodeling; PDGF-BB does not induce normal rat airway epithelial cells to product or release TGF-beta1 by phosphorylation of ERK. Glucocorticoids can inhibit phosphorylation of ERK. PMID- 17919387 TI - [Inhibition of the aquaporin-1 gene expression by RNA interference: experiment with cultured rat pleural mesothelial cells]. AB - OBJECTIVE: To investigate the influence of RNA interference (RNAi) on the expression of aquaporin-1 (AQP1) gene and to investigate the feasibility of gene therapy for pleural effusion. METHODS: Two recombinant plasmids with shRNAs targeting the AQP1 gene, AQP1-1-pGenesil and AQP1-2-pGenesil-1 were constructed. Pleural mesothelial cells were obtained from rats, cultured, and randomly divided into 5 groups: blank control group, Lipofectamine 2000 control group, HK negative control group, AQP1-1-pGenesil-1 transfected group, and AQP1-2-pGenesil-1 transfected group. RT-PCR and Western blotting were used to detect the mRNA and protein expression of AQP1. RESULTS: The mRNA expression levels of aquaporin-1 of the AQP1-1-pGenesil-1 and AQP1-2-pGenesil-1 transfected groups were inhibited by 83.5% and 90.9% respectively, and the protein expression levels of the AQP1-1 pGenesil-1 and AQP1-2-pGenesil-1 transfected groups were inhibited by 41.2% and 67.6% respectively. CONCLUSION: RNAi can successfully inhibit the expression of AQP1 and has the feature of sequence correlation of shRNA in the cultured rat pleural mesothelial cells. It may be used as a potential new approach for gene therapy of pleural effusion. PMID- 17919388 TI - [Effect of vascular endothelial growth factor 165 gene transfection on repair of bone defect: experiment with rabbits]. AB - OBJECTIVE: To investigate the influence of vascular endothelial growth factor (VEGF) 165 gene transfection on the repair of bone defect. METHODS: 38 New Zealand rabbits underwent resection of a segment 1 cm in length in bilateral radii filled with absorbable gelatin sponge. Dilated solution of the plasmid pcDNA3.1/VEGF165 was injected into the bone defect of one side and normal saline was injected into the contralateral bone defect. 1, 2, 4, 6, 8, and 12 weeks later X ray examination was conducted to observe the repair of bone defect, and then 5 rabbits were killed at each time points to take out the bone defects. HE staining was used to observe the bone repair. The levels of microvessel density (MVD) 1 and 2 weeks after the operation were observed. RT-PCR was used to detect the mRNA expression of VEGF in the bone defect. Based on the results of RT-PCR the tissue mRNA expression of VEGF65 was detected by real-time quantitative polymerase chain reaction (RQ-PCR). RESULTS: X-ray examination showed that there was no significant difference in the wound healing between the two group 1 week after the operation in all rabbits. Some callus could be seen in the experimental group 2 weeks after. Twelve weeks after the operation the reconstruction of bone cortex was completed. Similar process occurred in the control sides but more lately. The MVD level 7 days after of the experimental group was 47.0 +/- 7.5, significantly higher than that of the control group (42.2 +/- 6.4, t = 2.4519, P = 0.0179), and the MVD level 14 days after of the experiment group was 69.1 +/- 5.4, significantly higher than that of the control group (56.1 +/- 6.1, t = 8.0347, P = 0.0000). In the experimental group the mRNA expression amounts of VEGF165 could be found 1 week after, gradually increased and peaked 3 weeks after, then decreased, and became stable 6 weeks after. The mRNA expression amounts of VEGF165 in the control group were lower than those of the experimental group. CONCLUSION: Local application of PcDNA3.1/VEGF(165) vector promotes the expression of VEGF165, and enhances the quantity of the angiogenesis, extra cellular matrix and healing of bone defect. PMID- 17919389 TI - [Relationship between the expression of hypoxia-inducible factor-1alpha and chemotherapy response in esophageal squamous cell carcinoma]. AB - OBJECTIVE: To observe the relationship between expression of hypoxia-inducible factor (HIF)-1alpha and chemotherapy response and clinical outcome. METHODS: Platixal in combination with cisplatin was used in 48 patients with ESCC at advanced stage. platixal 175 mg/m(2), d1; cisplatin 80 mg/m(2), d 2, d 3, d 4, 21 days was one cycle. Chemotherapy response was evaluated. Specimens mentioned above comes from pathology department, the first affiliated hospital of Zhengzhou university. Immuno-histochemistry was used to examine the expression of HIF 1alpha protein in esophageal squamous cell carcinoma. 20 cases normal tissue of esophago was used as control group. chi(2) test and Kaplan-Merier was used to analysis data. RESULTS: HIF-1alpha immunoreactivity was recognized in both cytoplasm and nuclei. HIF-1alpha protein positive expression rate was 43.75% (21/48) in tumor samples, and was negative expression in normal tissue. Among 21 cases with positive expression of HIF-1alpha, there was none complete response case and 5 cases of partial responses, and the response rate was 23.81%; whereas in 27 cases with negative expression of HIF-1alpha, there were 8 cases of complete response and 11 cases of partial responses, and the response rate was 70.37%. There was significant difference between two groups (P < 0.05). In patients of HIF-1alpha positive expression, the median time to tumor progression was 2.0 months, and the median actuarial survival was 6 months, and survival rate was 13.3% in two years; Whereas in patients of HIF-1alpha negative expression, the median time to tumor progression was 5.0 months, and the median actuarial survival was 11.0 months, and survival rate was 42.2% in two years. There was significant difference between two groups (P < 0.05). CONCLUSION: HIF-1alpha protein expression by immunohistochemistry may be a useful indicator to prediction the chemotherapy response and clinical outcome for esophageal squamous cell carcinoma. PMID- 17919390 TI - [Study on distribution of neural stem cells in the brain of Alzheimer disease transgenic mice through caudal vein transplantation]. AB - OBJECTIVE: To observe whether neural stem cells (NSCs) can successfully permeate into the brain through the blood-brain barrier (BBB) of Alzheimer disease (AD) transgenic mice and explore the methods of distribution and migration. METHODS: NSCs were isolated from 12-day-old fetal mice, cultured, labeled with enhanced green fluorescent protein (eGFP) and then transplanted into 10 AD transgenic mice and normal mice as controls through caudal vein. The mice were killed 48 h, 1 w, 2 w, and 4 w after transplantation respectively. The brains of the mice were made into continual frozen sections, the distribution and migration of the eGFP labeled NSCs were studied under fluorescence microscope. RESULTS: At different time points after transplantation the eGFP-labeled NSCs were diffusely distributed in the brain: distributed around the blood vessels in the first 48 h, and then migrated gradually towards the hippocampus and cortex until 4 weeks later. There were no obvious abnormal complications occurring after transplantation. CONCLUSION: NSCs can successfully permeate into the brain through the BBB of AD transgenic mice, and migrate into the brain parenchyma gradually. PMID- 17919395 TI - [The GRADE system in taking clinical decisions and the elaboration of recommendations and clinical practice guidelines]. PMID- 17919391 TI - [Effect of 17beta-estradiol on myocardial inducible NOS and endothelial NOS activities after ischemia-reperfusion in rat heart model]. AB - OBJECTIVE: To study the effect of 17beta-estradiol (E(2)) on myocardial inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) activities of ischemia-reperfusion myocardium in rat model. METHODS: 40 Langendorff perfused hearts isolated from bilateral ovariectomy (OVX) rat were randomly divided into four groups: control group (Group C), in which hearts were reperfused ex vivo for 15 minutes before ischemia in rat; ischemia-reperfusion control (Group I-R), in which modified St. Thomas II cardioplegic solution was perfused to perform the ischemia-reperfusion; dissolved control group (Group D), in which 0.1% dimethyl sulfoxide (DMSO) was dissolved in cardioplegic solution; E(2) group (Group E), in which 0.1% DMSO and 5 micromol of E(2) were dissolved in cardioplegic solution. Myocardial iNOS and eNOS activities were detected before and after reperfusion. Creatine phosphokinase (CPK), lactic dehydrogenase (LDH) and nitric oxide (NO) of coronary flow were measured, and heart function was evaluated to observe the effect of E(2) on myocardial ischemia-reperfusion injury (MIRI). RESULTS: Myocardial eNOS activity declined (P < 0.01) and iNOS activity increased after ischemia-reperfusion (8.87 +/- 3.74 nmol/min/g in Group C, 15.83 +/- 2.42 nmol/min/g in Group I-R, 17.60 +/- 5.21 nmol/min/g in Group E; P < 0.01), moreover, iNOS activity was much higher in Group E (25.85 +/- 5.21 nmol/min/g, P < 0.01). NO production was lower in Group I-R and Group D (P < 0.05), and higher in Group E (30.96 +/- 4.91 micromol/L in Group C, 33.16 +/- 5.57 micromol/L in Group E; P < 0.01). CPK and LDH were lower in Group E (P < 0.05). Recovery of heart function was better in Group E (P < 0.05). CONCLUSIONS: E(2) can relieve the injury of MIRI and promote heart function recovery by increasing iNOS activity and NO production. PMID- 17919396 TI - [Cooperative research and family medicine]. PMID- 17919397 TI - [Barriers for the implementation of cognitive services in Spanish community pharmacies]. AB - OBJECTIVE: To identify and assess barriers for dissemination, implementation, and sustainability of different cognitive services in Spanish community pharmacies. DESIGN: Qualitative study through semi-structured interviews followed by a descriptive analysis. METHOD: Two groups of experts related to Spanish community pharmacy were chosen. One with 15 community pharmacists with a relevant professional activity, while the other group (n=18) was related to pharmacy strategists. RESULTS: The lack of university clinical oriented learning, lack of pharmacists' attitude towards change and some uncertainty over their professional future were identified as barriers at the pharmacists' level. In relation to pharmacy as an organization the lack of clear messages by their leaders and the small volume of Spanish pharmacies were identified as barriers. In the category of pharmacy profession, the current reimbursement system, the lack of university clinical education, and the lack of leadership by current representative organizations were the barriers found. The lack of real involvement by health authorities, the lack of knowledge about the objectives of pharmacy cognitive services, and the lack of demand of these services by patients where also identified as barriers. CONCLUSIONS: Finally, 12 barriers were identified and grouped into 6 categories. These barriers fit in with the barriers identified in other countries. PMID- 17919398 TI - [Is it really so difficult to introduce cognitive services into Spanish community pharmacy?]. PMID- 17919399 TI - [Patients' understanding of the dietetic and pharmacological treatment of hypercholesterolaemia and its relationship with the achievement of objectives in patients receiving statins: OPINA study]. AB - OBJECTIVE: To evaluate the understanding of patients receiving statins about the dietary and therapeutic measures taken to control their cholesterol and to relate this to the achievement of therapy objectives. DESIGN: Transversal, multi-centre study, conducted between March and November, 2004. SETTING: Three-hundred seventy six primary care and specialist doctors from all over the country. PARTICIPANTS: A total of 2347 patients, with an average age of 59 (12) and 48% women, being treated with statins. METHOD: Survey to evaluate understanding of lipid-lowering diet and pharmacological treatment. The percentage of patients achieving therapy objectives in line with ATP III guidelines was determined. RESULTS: Understanding of diet was adequate only for foods with negative effects on the concentration of cholesterol. Overall, understanding was better in the young, those with a higher level of education, those with hyperlipaemia originating in the family, those under treatment for longer, and in those attended in primary care or internal medicine clinics; it was worse in diabetic patients and in those with cardiovascular disease. Patients with more understanding of their treatment reached their therapy objectives to a greater extent. CONCLUSION: Patients receiving statins treatment have an adequate understanding of diet, but little information on pharmacological treatment. This understanding is associated with the achievement of therapy objectives. PMID- 17919400 TI - [Pilot study for the inclusion of the portfolio of the family medicine specialist book in training]. AB - BACKGROUND: The Spanish Family Medicine National Commission is proposing a new portfolio-type Specialist Training Book (STB). OBJECTIVE: To pilot its contents, structure, and implementation strategies. DESIGN: Cross-sectional, descriptive study. SETTING: A Provincial Family Medicine Teaching Unit. PARTICIPANTS: Twenty eight tutors and 36 residents. METHODS: For 9 months the participants conducted a training assessment on diverse areas of competence by means of tasks at work. Tutors recorded information on the quality of reflection achieved by residents and the tasks they performed by means of the card model proposed in the STB. Residents filled in an ad hoc survey. A univariate analysis of quantitative data was conducted. RESULTS: Thirty-three surveys were received from residents; 21 tutors handed in 67 evaluation reports (average: 3 per tutor). They dealt with all the areas of competence, particularly those of communication, teaching, and ethics. Tasks most used were clinical sessions, critical incidents and video recording. Both tutors and residents thought that the new method could be useful for reflecting on clinical practice, understanding their own areas of competence better and for strengthening the tutor-trainee relationship, especially if some suggestions to improve its practical use and reduce time and effort involved were taken into account. CONCLUSIONS: The new STB in its current version or with some modifications is a useful tool for residents' training assessment and is probably accepted well in our ambit. PMID- 17919401 TI - [Evaluative study of medical-care costs in primary care]. AB - OBJECTIVE: To estimate the real costs of medical care by diagnostic groups at a primary care centre. DESIGN: Descriptive, retrospective study, based on the review of computerized medical records. SETTING: Urban primary care centre (PCC). PARTICIPANTS: All patients who attended the PCC during 2005. MAIN MEASUREMENTS: Mean medical care cost per visit in euros, broken down for health professionals, diagnostic procedures and drugs costs, and stratified by diagnostic groups. RESULTS: The most frequent visits were for pulmonary, locomotor, cardiovascular, and gastro-intestinal conditions. The mean number of visits per patient attended was 8.7 (SD, 9.4); and per patient registered at the centre, 5.9 (8.7). The highest costs were for cardiovascular (18.96%; 95% CI, 18.93%-18.99%), locomotor (11.21%; 95% CI, 11.18%-11.23%), psychological (10.69%, 95% CI, 10.66%-10.71%), pulmonary (10.20%; 95% CI, 10.17%-10.22%) and endocrinal-nutritional (9.61%; 95% CI, 9.58%-9.63%) problems. Drugs expenditure accounted for 65% of the total cost; visits to health professionals, for 33%; and procedures, for 2%. Overall cost per inhabitant was 239.1 (493.6) euros, and per patient attended was 349.5 (563.5). CONCLUSIONS: Cardiovascular disease conditions are much the most costly ones in terms of overall medical cost. Psychological conditions are located in second place in terms of pharmaceutical cost; and in third place, in terms of overall medical-care cost. PMID- 17919402 TI - [The bioethics portfolio in family doctor training]. PMID- 17919403 TI - [Current situation of family medicine in Venezuela]. PMID- 17919404 TI - [Proposal for a map of methods and techniques to help tutors to "travel through the teaching programme"]. PMID- 17919405 TI - [White-coat hypertension indicators diagnosed through self-measurement of blood pressure at home]. PMID- 17919406 TI - [Cover with conjugated heptavalent pneumoccal vaccine in the child population of Tarragona-Valls, Spain]. PMID- 17919407 TI - [Control of vascular risk factors before and after an AMI or a CVA]. PMID- 17919408 TI - [Interns and the pharmaceutical industry]. PMID- 17919409 TI - [Knowledge of contraception and transmission of HIV and hepatitis B in the sub Saharan immigrant population]. PMID- 17919410 TI - [Little relevant news on secondary prevention of breast cancer]. PMID- 17919412 TI - [Nonrespiratory effects of smoking]. PMID- 17919411 TI - [Mediastinoscopy: an endangered species?]. PMID- 17919413 TI - [Expression of proteins associated with multidrug resistance and resistance to chemotherapy in lung cancer]. AB - OBJECTIVE: Membrane transporters are proteins that play a crucial role in resistance to chemotherapy. The aim of this study was to assess the influence of membrane transporter protein expression on chemotherapeutic response. MATERIAL AND METHODS: One hundred and forty seven samples of tumor tissue were collected from 143 patients; 35 samples were obtained by bronchoscopy and 112 were surgical specimens. A total of 101 samples from 99 patients were adequate for study. Cryopreserved samples were subjected to immunohistochemical analysis to detect 3 proteins associated with multidrug resistance: P-glycoprotein (Pgp), multidrug resistance-associated protein 1 (MRP1), and lung resistance protein (LRP). RESULTS: In 16 cases none of the proteins were expressed. A single protein was expressed in 32 (3 Pgp, 11 MRP1, and 18 LRP); 2 in 34 cases (24 Pgp and LRP; 5 MRP1 and Pgp; 5 MRP1 and LRP); and all 3 in 17 cases. No significant relationship was found between age and the expression of Pgp (P=.74), MRP1 (P=.95), or LRP (P=.26). Nor were there significant differences in number (P=.72) or type of coexpressed proteins (P=.39) by sex, by tumor stage (number, P=.55; type, P=.21), or by tumor grade (number, P=.59; type, P=.51). There was a highly significant trend toward coexpression of Pgp and LRP (P< .01) but not of Pgp and MRP1 (P=.18) or MRP1 and LRP (P=.26). MRP1 was expressed less often in adenocarcinoma. LRP was expressed less often in squamous cell carcinoma than in adenocarcinoma and undifferentiated large cell carcinoma. Coexpression of Pgp, MRP1, and LRP was observed most often in squamous cell carcinoma. CONCLUSIONS: Proteins associated with multidrug resistance are commonly expressed in lung cancer. Of the 3 proteins studied, LRP was the one most often found. Coexpression of more than 1 of the proteins was found in a considerable percentage of patients. Pgp was mainly found to be coexpressed with LRP. Pgp expression and the number of coexpressed proteins seemed to have a negative impact on response to chemotherapy. PMID- 17919414 TI - [Reference values for inspiratory capacity in healthy nonsmokers over age 50 years]. AB - OBJECTIVE: The role of dynamic hyperinflation in triggering dyspnea and limiting exercise capacity in patients with chronic obstructive pulmonary disease has been recognized in recent years. The degree of dynamic hyperinflation can be assessed by measuring reduction in inspiratory capacity (IC). The aim of this study was to establish reference values for IC in healthy individuals of both sexes between the ages of 50 and 87 years, as such data are scarce in the literature. SUBJECTS AND METHODS: We studied 155 healthy volunteers (93 women) with normal spirometry. None had a prior history of respiratory, cardiovascular, or systemic diseases that might alter lung function. All were never-smokers. IC was measured during a normal, unforced inspiration to total lung capacity starting from functional residual capacity. The highest value of 6 satisfactory maneuvers was recorded. Sex, height, age, and weight were included in the regression equations. One thousand bootstrap samples for each sex were also analyzed. RESULTS: For each sex, we found that a model including age, height, and weight produced IC prediction equations with a coefficient of determination (r2) of 0.414 for women and 0.447 for men. The mean (SD) intrasubject coefficient of variation was 4.3% (2%) for IC measured during a single session and 5.1% (0.4%) for measurements from 5 weekly sessions. CONCLUSIONS: Our results provide reference equations for IC that are valid for a healthy population over 50 years of age. Predicted values were similar to those recently obtained in an Italian population aged between 65 and 85 years. PMID- 17919415 TI - [Prognostic value of transthoracic echocardiography in hemodynamically stable patients with acute symptomatic pulmonary embolism]. AB - OBJECTIVE: To determine the prognostic value of transthoracic echocardiography in hemodynamically stable patients diagnosed with acute symptomatic pulmonary embolism. PATIENTS AND METHODS: Hemodynamically stable outpatients diagnosed with acute symptomatic pulmonary embolism at a tertiary university hospital were prospectively included in the study. All patients underwent transthoracic echocardiography within 48 hours of diagnosis. The primary endpoint was all-cause mortality at 1 month. RESULTS: Right ventricular dysfunction was documented by echocardiography in 86 of the 214 patients (40%) in our series. In the first month of follow-up, 7 patients died--4 with positive echocardiographic findings and 3 with negative findings (odds ratio, 2.0; 95% confidence interval, 0.4-9.3; P=.41). For the primary endpoint, the negative predictive value of transthoracic echocardiography was 98%, the positive predictive value was 5%, and the negative likelihood ratio was 0.7. The negative predictive value was 100% and the positive predictive value was 3% when we analyzed death due to pulmonary embolism only. CONCLUSIONS: In our setting, transthoracic echocardiography is not useful for prognostic stratification of hemodynamically stable patients with pulmonary embolism. PMID- 17919416 TI - [Clinical utility of transbronchial needle aspiration of mediastinal lymph nodes in the diagnosis of sarcoidosis in stages I and II]. AB - OBJECTIVE: To analyze the individual and combined diagnostic yield and clinical utility of transbronchial needle aspiration and transbronchial biopsy in the histologic diagnosis of sarcoidosis in stages I and II. PATIENTS AND METHODS: We performed a prospective study enrolling all the patients admitted to our hospital between July 2001 and June 2006 with mediastinal lymph nodes of a diameter of over 10 mm and clinical and radiological indication of sarcoidosis. The patients underwent the following tests in the order specified: a transbronchial needle aspiration of 1 or more lymph nodes using a 19-gauge histology needle until 2 satisfactory specimens were obtained, a bronchoalveolar lavage, and 4 to 6 transbronchial biopsies. RESULTS: A total of 32 patients were enrolled during the study period. Of these, 26 were finally diagnosed with sarcoidosis. The mean (SD) age of the patients was 38.9 (10.6) years and there were 13 women (50%). The mean (SD) lymph node diameter was 23.5 (6.5) mm. The most common lymph node aspiration sites were the subcarinal station (9 patients, 34.6%) and a combination of the subcarinal and paratracheal stations (7 patients, 26.9%). Computed tomography scans of the chest revealed parenchymal involvement in 12 (46.2%) patients. Fifteen (57.7%) patients were diagnosed by transbronchial biopsy and 16 (61.5%) by transbronchial needle aspiration. These techniques were used in isolation in 7 (26.9%) and 8 (30.8%) patients, respectively (P>.05). Diagnosis was achieved with the combined results of transbronchial biopsy and needle aspiration in 23 (88.4%) patients. Overall, the diagnostic yield of the combined techniques was significantly higher than that of either one in isolation (P=.01 vs transbronchial biopsy and P=.02 vs needle aspiration). In patients with parenchymal involvement, yield was better for the combined techniques only in comparison with transbronchial biopsy (P=.01). Only 4 (15%) patients developed complications and 20 (76.9%) tolerated the procedures well. CONCLUSIONS: Diagnostic yield was significantly higher when transbronchial biopsy and transbronchial needle aspiration were used in combination than when used alone, particularly in patients without parenchymal involvement. There were few complications and tolerance was good. PMID- 17919417 TI - [Experimental models for the study of pulmonary fibrosis: current usefulness and future promise]. AB - Diffuse interstitial lung diseases form a group of respiratory diseases about which many questions remain to be answered. In recent years there have been major advances in the correct diagnostic classification of each disease, and therefore, the essential foundations have been laid for investigation of their pathophysiology. However, both the triggers and the precise mechanisms that lead to irreversible changes in the lung parenchyma remain to be identified. Idiopathic pulmonary fibrosis is the most common diffuse interstitial lung disease and has the worst prognosis. Current treatments are empirical and the response is random; furthermore, they do not improve survival. Consequently, most basic research has focused on the pathophysiology of the disease and on identifying an effective therapeutic approach. The aim of this review is to describe the experimental studies that have begun to open the way towards an understanding of the complex process of fibrosis. PMID- 17919418 TI - [Lung cancer articles in Archivos de Bronconeumologia: 2 years on from lung cancer year of the Spanish Society of Pulmonology and Thoracic Surgery (SEPAR)]. PMID- 17919419 TI - [Accelerated phase of idiopathic pulmonary fibrosis]. AB - The natural history of idiopathic pulmonary fibrosis is characterized by a slow progression resulting in respiratory failure and death. The progression to the fulminant form is rapid in a small percentage of cases, however. Within weeks or months, patients develop respiratory distress, and extensive ground-glass patterns can be seen in computed tomography scans and hyaline membranes in biopsy samples. This is described as an accelerated phase of idiopathic pulmonary fibrosis, in which elevated levels of acute-phase reactants and tumor markers have been reported. To date, the monoclonal tumor marker, CA 15/3 has not been associated with the accelerated phase. PMID- 17919420 TI - [Early pleuropulmonary toxicity associated with cabergoline, an antiparkinsonian drug]. AB - We report a case of pleural effusion, pericardial thickening, and pulmonary involvement in a patient with dry cough, dyspnea, edema, and changes in the skin of the lower limbs. Treatment with cabergoline (Sogilen) had been started 4 months earlier. Pleural effusion, pericardial thickening, and impaired pulmonary function (airflow obstruction, increased airway resistance, and reduced carbon monoxide diffusing capacity) were observed. The Naranjo scale pointed to a probable relationship between cabergoline and these adverse effects. We report on outcome after 2 months of follow-up, during which time there was a slow and incomplete improvement in respiratory function. This is the first case in our practice setting of early pleuropulmonary toxicity associated with cabergoline. PMID- 17919421 TI - [Rebuttal of: "Can home prophylaxis for venous thromboembolism reduce mortality rates in patients with chronic obstructive pulmonary disease?"]. PMID- 17919422 TI - [Uric acid and its relationship to creatinine levels and hypoxia]. PMID- 17919423 TI - [Evaluation of regular physical activity in COPD patients with an accelerometer and a questionnaire: a pilot study]. PMID- 17919424 TI - [Sexually transmitted diseases and immigration in Spain]. PMID- 17919425 TI - [The Cochrane Skin Group]. AB - Over the last 10 years, the advent of evidence-based medicine has caused a paradigm shift in medical thinking and clinical practice. The Cochrane Skin Group organizes, drafts, publishes, and updates systematic reviews of randomized clinical trials of therapeutic interventions in skin diseases to assist in evidence-based clinical decision making. This article provides background information on the history and operation of this group, including the process by which systematic reviews are prepared. PMID- 17919426 TI - [Corticosteroids and osteoporosis]. AB - Corticosteroids are the principal cause of secondary osteoporosis due to drug treatment. Doses of more than 5 mg daily and periods of treatment lasting more than 3 months increase the risk of osteoporosis and fragility fractures. It is therefore essential to be aware of measures to reduce the risk of osteoporosis in our patients. PMID- 17919427 TI - [Longitudinal study of different metastatic patterns in the progression of cutaneous melanoma]. AB - INTRODUCTION: Compared with other tumors, melanoma has displayed one of the largest increases in incidence in recent years, and it is known to have a high metastatic potential. In cases of metastasis, approximately two-thirds of patients have lymph-node metastases and one-third develop systemic metastases. However, few studies have been reported that analyzed different metastatic patterns according to the natural history of melanoma. The main aim of this study was to analyze the different metastatic pathways and patterns and to assess the time course of development of metastases from cutaneous melanoma. MATERIAL AND METHODS: A retrospective study was performed in 575 patients with onset of primary melanoma between 1990 and 2004. During follow-up, 67 patients developed metastases. Different pathways for metastasis were established and evaluated. We identified 4 metastatic pathways according to the metastatic pattern during progression of the melanoma. The time course of metastases was also evaluated. Finally, we analyzed melanomas with local recurrence in terms of whether or not systemic progression occurred. RESULTS: Melanoma metastases first occurred in local lymph nodes in 55.2 % of the patients. Initial metastasis was systemic in 14.9 % of the patients. The anatomical location and tumor thickness influenced which metastatic pathway was followed. Distant metastases occurred after a mean of 25 months regardless of the pathway followed CONCLUSIONS: The development of distant metastases displays a constant time course and the time to onset is independent of the metastatic pathway. This observation may explain why sentinel lymph node biopsy has a limited impact on overall survival of melanoma patients. PMID- 17919428 TI - [Analysis of lymphocyte populations in psoriatic plaques following inhibition of tumor necrosis factor alpha with etanercept]. AB - INTRODUCTION: Psoriasis is a disease with a strong immunological component in which there is a predominant T helper 1 cell-mediated immune response. Etanercept, a receptor for tumor necrosis factor a that blocks its action, is a new drug with proven efficacy in the treatment of psoriasis. OBJECTIVES: The primary objective of this study was to assess the histological response to etanercept by analyzing the lymphocyte populations in psoriatic plaques. The secondary objectives were to assess the clinical response to the drug using the Psoriasis Area and Severity Index (PASI) and to analyze the effect of etanercept on peripheral blood lymphocyte populations. METHODS: Ten patients with plaque psoriasis and a PASI score greater than 10 were included in the study. A clinical assessment was performed in all patients along with a 4-mm skin punch biopsy of a plaque and analysis of peripheral blood lymphocyte populations at baseline and after 12 weeks of etanercept therapy at a dose of 50 mg per week. RESULTS: There was a significant reduction in different lymphocyte populations in the plaques following treatment with etanercept. The mean (SD) number of CD4+ T lymphocytes per microscopic field decreased from 16.93 (8.13) at baseline to 6.51 (3.46) after treatment with etanercept (P < 007). CD8+ T lymphocytes also decreased from 17.73 (9.77) before treatment to 10.50 (9.4) after treatment (P < 005). An overall improvement in PASI score was also observed: 33.30 (10.71) at baseline versus 15.20 (13.28) following treatment (P < 008). Nine out of 10 patients showed improvement. No significant differences were observed in peripheral blood lymphocyte populations before and after treatment. CONCLUSIONS: Etanercept leads to clinical improvement of psoriasis and reduces inflammatory infiltration of the lesions without affecting peripheral blood lymphocyte populations. PMID- 17919429 TI - [Stewart-Bluefarb syndrome]. AB - Stewart-Bluefarb syndrome is a rare condition involving skin lesions that share clinical features with Kaposi sarcoma and that are secondary to an underlying arteriovenous fistula. We report the case of a 24-year-old man with progressive growth of skin lesions on the lower third of his left leg. Diagnosis of Stewart Bluefarb syndrome was confirmed histologically and with detection of an arteriovenous fistula. PMID- 17919430 TI - [Lupus erythematosus panniculitis presenting as palpebral edema and parotiditis]. AB - Lupus erythematosus panniculitis or lupus erythematosus profundus is characterized by inflammation of the deep dermis and subcutaneous tissue. It can occur in isolation or associated with chronic systemic or discoid lupus erythematosus. It usually consists of nodules and hardened subcutaneous plaques on the forehead, cheeks, proximal extremities, and buttocks. Periorbital and parotid involvement are rare and can lead to misdiagnosis. We present the case of a patient with lupus erythematosus panniculitis who presented with palpebral edema and involvement of the periocular fat and parotid gland. PMID- 17919431 TI - [Kaposi sarcoma associated with systemic corticosteroid therapy]. AB - Kaposi sarcoma (KS) was first described in 1872 by Moritz Kaposi. Its epidemiology is suggestive of an infectious disease and in 1994 Chang and coworkers identified DNA sequences corresponding to a previously unidentified herpes virus--human herpes virus 8 (HHV-8)--in AIDS-associated KS biopsies. It is now believed that the presence of HHV-8 is a necessary condition but not sufficient on its own to cause KS. Other factors such as immunosuppression should also be considered and it is known that immunosuppressive therapy increases the risk of KS. We describe a patient who developed KS after prolonged prednisone therapy for temporal arteritis. PMID- 17919432 TI - [Topical chemotherapy for actinic keratosis and nonmelanoma skin cancer: current options and future perspectives]. AB - Actinic keratosis is currently considered not to be a precursor of squamous cell carcinoma but, rather, an initial stage of the disease. Furthermore, the incidence of both squamous cell carcinoma and basal cell carcinoma continues to increase. Topical drugs are now becoming widely used in the therapeutic management of nonmelanoma skin cancer and its precursor lesions. Here, we review the various topical drugs that are currently available and discuss their advantages and drawbacks. Therapeutic options include retinoids, 5-fluoracil, diclofenac, imiquimod, and photodynamic therapy. PMID- 17919433 TI - [Reticular telangiectatic plaque]. PMID- 17919434 TI - [Palmoplantar hyperkeratotic plaques]. PMID- 17919436 TI - [Medicine and disease in baroque art]. PMID- 17919435 TI - [Dry gangrene of the foot]. PMID- 17919437 TI - [Medicine in baroque painting]. PMID- 17919438 TI - [Generalized syringoma: a case study]. PMID- 17919439 TI - [Psoriasis at the site of healed herpes zoster: Wolf's isotopic response]. PMID- 17919440 TI - [Malignant degeneration of linear hidroacanthoma simplex]. PMID- 17919441 TI - [Hypersensitivity to the antioxidant ethoxyquin]. PMID- 17919442 TI - Effect of time of day on arithmetic fact retrieval in a number-matching task. AB - The goal of this study is aimed to investigate a possible effect of time of day on arithmetic fact retrieval in a number-matching task. We tested 96 students (age range 19-33 years) at 9 a.m. and at 1 p.m., in a counterbalanced order. The subjective alertness levels were also recorded. As regards retrieval efficiency, the results showed that the sum interference effect was significantly more pronounced in the morning (9 a.m.) than at midday (1 p.m.). As expected, participants showed higher subjective alertness levels at 1 p.m. than at 9 a.m.. However, the difference of subjective alertness did not completely explain differences in interference at two testing sessions. The results could be explained with respect to the variation of working memory efficiency during the day. PMID- 17919443 TI - Successful vaccination against Leishmania chagasi infection in BALB/c mice with freeze-thawed Leishmania antigen and Corynebacterium parvum. AB - This study evaluated the potential of a Leishmania antigen vaccine in protecting BALB/c mice against Leishmania chagasi. Mice received two subcutaneous doses of L. amazonensis vaccine with Corynebacterium parvum and subsequent boost was done without adjuvant. One week later, mice were challenged with L. chagasi. We observed that this vaccine caused a significant reduction in parasite load in liver and spleen and induced a high production of IFN-gamma and IL-4 by spleen cells from vaccinated mice in response to Leishmania antigen. Together, our data show that this vaccine is capable of inducing a Th1/Th2 response that is important to control parasite replication. PMID- 17919444 TI - Water adsorption on carbons--critical review of the most popular analytical approaches. AB - The purpose of the current study is to present the state of art in the field of analytical description of water sorption on carbons. We discuss the most important and promising models proposed recently (for example by Mahle; Talu and Meunier; and Malakhov and Volkov) as well as some older theoretical models inspired by the pioneering ideas proposed in the papers of Dubinin, Serpinsky, Barton, D'Arcy, Watt, Do and Do and others. The applicability, advantages, and defects of all these analytical formulas are pointed out and some new approaches in this field are presented. The special attention is paid to the finite adsorption space and the possible involvement of partial chemisorption, i.e. the existence of various types of the hydrophilic centres. Since the calculation of isosteric enthalpy from an adsorption equation, and the comparison of theoretical enthalpy plot with the values measured calorimetrically, is the fundamental condition for the verification of the correctness of an adsorption model, for all considered models we show the corresponding adsorption enthalpy equations. The validity of all mentioned above models is verified for the data measured for five water-activated carbon systems. Finally, a summary of obtained results and some perspectives and suggestions for the description of experimental data are presented. From the analysis of experimental data it is seen that developed recently- the heterogeneous Do and Do model is probably the most successful for the simultaneous description of water adsorption and enthalpy of adsorption results. PMID- 17919445 TI - Reproducibility of visual field end point criteria for standard automated perimetry, full-threshold, and Swedish interactive thresholding algorithm strategies: diagnostic innovations in glaucoma study. AB - PURPOSE: To compare the interthreshold and intrathreshold strategy agreement of visual field end point criteria for standard automated perimetry (SAP) with the full-threshold (FT) algorithm and the Swedish interactive threshold algorithm (SITA). DESIGN: Prospective, longitudinal cohort study. METHODS: The interstrategy group included a randomly selected eye of 173 participants in the Diagnostic Innovations in Glaucoma Study who had undergone FT algorithm and SITA analysis within three months (sequence 1, FT + SITA). Intrastrategy agreement for the FT algorithm (sequence 2, FT + FT) was tested for 44 (25.4%) participants who had undergone FT analysis within one year of the FT used in the interstrategy pairing, and for 89 patients (51.4%) who had undergone SITA analysis within one year before (sequence 3, SITA + SITA). Four different end point criteria using Statpac II indices were tested. Interstrategy agreement was compared with intrastrategy agreement using kappa statistics. RESULTS: FT + SITA agreement (kappa) for pattern standard deviation (PSD) < 1% was 0.82; for PSD < 5%, the kappa value was 0.64; and for four or more pattern deviation plot points, the kappa value was 0.43. Agreement with glaucoma hemifield test (GHT) results was significantly higher (P < .01) for FT + FT (kappa = 0.94) than FT + SITA (kappa = 0.67), and approached significance (P = .07) when comparing FT + FT with SITA + SITA (kappa = 0.77). GHT results were more likely to be abnormal on the SITA analysis than on the FT analysis. No other significant differences were found. CONCLUSIONS: To minimize misinterpreting abnormal GHT results on SITA as evidence of change when switching strategies, both SITA and FT should be performed and compared within a short period. Other indices are comparable between the two strategies. PMID- 17919446 TI - Association between intraocular pressure variation and glaucoma progression: data from a United States chart review. AB - PURPOSE: To evaluate whether greater intraocular pressure (IOP) variation between visits was associated with higher likelihood of glaucoma progression. DESIGN: Cohort study. METHODS: A five-year minimum of data (June 1, 1990 through January 22, 2002) was collected on 151 patients (302 eyes) from 12 United States specialty centers. A post hoc analysis of visual field (VF) progression, glaucoma medication, intraocular pressure (IOP), and other ocular data was conducted for two nonmutually exclusive cohorts based on retrospective data abstracted well after actual patient visits. Mean IOP and standard deviations (SD) were calculated before treatment (medication or surgery) or progression, whichever occurred first, and before progression regardless of treatment. IOP variables were assessed in a univariate fashion; Cox proportional hazards models evaluated glaucoma progression as an outcome measure and IOP SD as a main predictor, controlling for covariates. RESULTS: In cohort 1 (55 patients; 84 eyes), mean age was 63 years (range, 37 to 85 years), 58% were female, and 19% of eyes underwent VF progression. In cohort 2 (129 patients; 251 eyes), mean age was 66 years (range, 19 to 88 years), 55% were female, and 27% of eyes underwent VF progression. Mean IOP was 16.5 mm Hg (IOP SD, 2.0 mm Hg), and 16.4 mm Hg (IOP SD, 2.7 mm Hg) in cohorts 1 and 2, respectively. Controlling for age, mean IOP, VF stage, and other covariates, each unit increase in IOP SD resulted in a 4.2 times and 5.5 times higher risk of glaucoma progression for cohort 1 (95% confidence interval [CI], 1.3 to 12.9) and cohort 2 (95% CI, 3.4 to 9.1), respectively. CONCLUSIONS: IOP variability is an important predictor of glaucoma progression; SD is a convenient measure of variability to assess glaucoma progression risk. PMID- 17919447 TI - Corneal endothelial morphologic features in cataract and clear lens in an Indian population. AB - PURPOSE: To compare the morphologic features of corneal endothelium in patients with cataract and clear lenses. DESIGN: A single, observational clinic-based case control study. METHODS: In this single, observational clinic-based case-control study at Iladevi Cataract & IOL Research Centre, Ahmedabad, India, 2,025 eyes of 2,025 individuals with cataract and clear lenses were evaluated for corneal endothelial morphologic features, which included the endothelial cell density (ECD), coefficient of variation (CV) of cell area, and percentage of hexagonal cells. Nine hundred and thirty-nine eyes of patients with clear lenses were assigned to the control group and 1,086 eyes of patients with cataract were assigned to the case group. The main outcome measures studied were the influence of variables such as age, gender, axial length, and anterior chamber depth on the corneal endothelial morphologic features in patients with cataract and clear lenses. RESULTS: Eyes in the cataract group had a low mean ECD (2241.52 +/- 158.69 cells/mm(2) vs 2446.04 +/- 197.79 cells/mm(2)), high CV of cell area, and low mean percentage of hexagonal cells compared with those in the clear lens group. Multiple linear regression analysis revealed a reduction in ECD in the presence of cataract, in female subjects, and with advancing age, whereas an increase in ECD was noted with an increase in anterior chamber depth of more than 2.7 mm (P = .020) and an increase in axial length of more than 24.0 mm (P < .001). CONCLUSIONS: Patients with cataract had lower levels of ECD than the corresponding normal population. In multiple regression analysis, after taking into account all the variables, we found a significant impact on the morphologic features of corneal endothelium in patients with cataract when compared with those with clear lenses. PMID- 17919449 TI - Quantitative measurements of vancomycin binding to self-assembled peptide monolayers on chips by quartz crystal microbalance. AB - This paper describes direct binding of a small vancomycin to peptide ligands immobilized on a sensor chip using quartz crystal microbalance. In this study, the binding ligands were composed of three components: a molecular recognition element (peptide), a conformationally flexible and hydrophilic linker, and a long chain alkanethiol. These peptide ligands were used to establish the well-packed, self-assembled monolayers on quartz chips and could be readily synthesized using conventional organic chemistry protocols. Results of quartz crystal microbalance measurements showed that vancomycin specifically associated with the d-Ala-d-Ala containing peptide with an affinity of 3.2+/-0.3 microM and was, as expected, completely inactive to the self-assembled monolayer presenting l-Ala-l-Ala peptide. The dissociation constant obtained correlated well with values reported in literature and was further confirmed by surface plasmon resonance measurement (2.7+/-0.7 microM). The technique used in this study should be applicable to both peptidyl and nonpeptidyl ligands of greater complexity than that used here. This method is practical, it provides quantitative binding information, and complicated analysis is avoided. PMID- 17919448 TI - Refinement of a radioreceptor binding assay for nicotinic acid adenine dinucleotide phosphate. AB - The measurement of changes in nicotinic acid adenine dinucleotide phosphate (NAADP) levels in cells has been, and remains, key to the investigation of the functions of NAADP as a Ca2+ -releasing second messenger. Here we provide details of how to isolate NAADP from cells by extraction with perchloric acid and then measure the NAADP using a radioreceptor assay. We demonstrate that NAADP is neither generated nor broken down during sample processing conditions and that radioreceptor assay is highly selective for the detection of NAADP under cell extract conditions. Furthermore, a number of improvements, such as solid-state detection of the radioactivity, are incorporated to enhance the safety of the procedure. Finally, we have developed a new method to prevent the endogenous metabolism of NAADP by chelating Ca2+ with bis-(o-aminophenoxy)ethane-N,N,N',N' tetraacetic acid (BAPTA), thereby reducing the difficulty of catching a small transient rise in NAADP levels. In summary, we have refined and improved a method for measuring NAADP levels and presented it in a manner accessible to a wide range of laboratories. It is expected that this will enhance research in the NAADP field. PMID- 17919451 TI - Unraveling the components of protein translocation pathway in human malaria parasite Plasmodium falciparum. AB - The targeting and translocation of proteins is an essentially required and conserved process in all the living organisms. This complex process involves multiple steps and requires a variety of factors before the protein reaches its final destination. The major components of translocation machinery are signal recognition particle (SRP) and secretory (Sec) complex. These are composed of highly conserved components. SRP contains SRP RNA and other polypeptides such as SRP9, SRP14, SRP19 and SRP54. Sec complex is composed of Sec61alphabetagamma, Sec62 and Sec63. In this review using bioinformatics approach we have shown that the P. falciparum genome contains the homologues for all of these and other factors such as SRP receptor, and TRAM (translocation associated membrane protein), which are required for post- and co-translational protein translocation. We have also shown the various steps of translocation in a hypothetical model. PMID- 17919450 TI - Phospholipid-protein coatings for chiral capillary electrochromatography. AB - A phospholipid-bovine serum albumin (BSA) coating was developed for chiral capillary electrochromatographic separation of d- and l-tryptophan. Temperature, liposome composition, and liposome-BSA mixing and extrusion were found to have critical effects on the chiral separation of d- and l-tryptophan in terms of resolution, separation efficiency, and migration times. A solution of 0.5mM phosphatidylcholine (PC)-1 mg/ml BSA performed better than a solution of 0.5mM PC/phosphatidylserine (PS) (80:20, mol%)-1 mg/ml BSA as capillary coating; baseline separation of the enantiomers with satisfactory resolution was then achieved. Temperature played a crucial role in the chiral separation, as demonstrated for phospholipid-coated capillaries immobilized with BSA and lysozyme. The d- and l-tryptophans showed a marked difference in separation efficiency on the PC-BSA-coated capillary; the theoretical plate number of l tryptophan was above 500,000 m(-1), whereas that of d-tryptophan was only about 22,000 m(-1). Immobilized BSA (pI 4.7) showed better chiral separation selectivity for the enantiomers than did immobilized lysozyme (pI 10.5), alpha chymotrypsin (pI 8.1-8.3), or avidin (pI 10.0-10.5); also resolution was better and analysis time was faster. Hydrophobic interactions played an important role in the BSA-immobilized phospholipid-coated capillaries. The importance of protein net charge and molar mass for its immobilization in phospholipid-coated capillaries is discussed. PMID- 17919452 TI - Effect of unsaturated bonds in the sn-2 acyl chain of phosphatidylcholine on the membrane-damaging action of Clostridium perfringens alpha-toxin toward liposomes. AB - Clostridium perfringens alpha-toxin degrades phosphatidylcholine (PC) in the bilayer of liposomes and destroys the membrane. The effect of the type and position of unsaturation in the fatty acyl chain of PC (18:0/18:1 PC) synthesized on the toxin-induced leakage of carboxyfluorescein (CF) from PC liposomes was examined. Differential scanning calorimetry showed that the phase transition temperature (T(m)) was minimal when the triple bond was positioned at C (9) in the sn-2 acyl chain. The toxin-induced CF leakage decreased with the migration of the bond from C (9) to either end of the acyl chain in PC. The PC containing the cis-double bond had a similar T(m) to that with the triple bond, but a lower value than the PC containing the trans-double bond. Furthermore, the toxin induced leakage from liposomes composed of PC containing the cis-double bond resembled that with PC having the triple bond and was greater than that from liposomes with PC having the trans-double bond. The binding of a H148G mutant to PC liposomes showed a reciprocal relationship in terms of the T(m) value of PC containing the triple bond. These results indicate that the toxin-induced membrane damage is closely related to membrane fluidity in liposomes. PMID- 17919453 TI - Dynein light chain 1 peptide inhibits human immunodeficiency virus infection in eukaryotic cells. AB - Human immunodeficiency virus (HIV) uses kinases such as Pak1 and macropinocytosis for a productive infection. Recently dynein light chain 1 (DLC1), a component of the dynein motor, was identified as a Pak1 substrate and interacted with the C terminal region of DLC1 (aa 61-89). The dynein motor is implicated in retrograde transport, also of HIV, to the nucleus. It is known that DLC1 is important in macropinocytosis, and anti-dynein antibodies inhibit a productive HIV infection. Here, we show that in Hela-beta-gal cells macropinocytosis was effectively blocked by a peptide spanning the C-terminal 19 amino acids of DLC1. We also found that the DLC1 peptide was capable of inhibiting the early entry steps of HIV, and the DLC1 peptide efficiently inhibited a productive HIV infection, and cooperated with the anti-HIV activity of CD4 antibodies. Taken together, the potentially therapeutic DLC1 peptide represents an interesting class of HIV inhibitors, targeting an essential cellular component for HIV infection. Our findings raise the possibility that the use of a DLC1 peptide in combination with currently used anti-HIV agents, might offer additional arsenal against HIV infection in human cells. PMID- 17919454 TI - Modulation of Spc1 stress-activated protein kinase activity by methylglyoxal through inhibition of protein phosphatase in the fission yeast Schizosaccharomyces pombe. AB - Methylglyoxal, a ubiquitous metabolite derived from glycolysis has diverse physiological functions in yeast cells. Previously, we have reported that extracellularly added methylglyoxal activates Spc1, a stress-activated protein kinase (SAPK), in the fission yeast Schizosaccharomyces pombe [Y. Takatsume, S. Izawa, Y. Inoue, J. Biol. Chem. 281 (2006) 9086-9092]. Phosphorylation of Spc1 by treatment with methylglyoxal in S. pombe cells defective in glyoxalase I, an enzyme crucial for the metabolism of methylglyoxal, continues for a longer period than in wild-type cells. Here we show that methylglyoxal inhibits the activity of the protein phosphatase responsible for the dephosphorylation of Spc1 in vitro. In addition, we found that methylglyoxal inhibits human protein tyrosine phosphatase 1B (PTP1B) also. We propose a model for the regulation of the activity of the Spc1-SAPK signaling pathway by methylglyoxal in S. pombe. PMID- 17919455 TI - Marine natural products as targeted modulators of the transcription factor NF kappaB. AB - The inducible transcription factor nuclear factor-kappaB (NF-kappaB) plays an important role in the regulation of immune, inflammatory and carcinogenic responses. While normal NF-kappaB activation is necessary for cell survival and immunity, deregulated NF-kappaB expression is a characteristic phenomenon in cancer development, as well as in several inflammatory diseases. Hence, NF-kappaB has become a major target in drug discovery, and several natural and synthetic compounds have been investigated for their potential to inhibit NF-kappaB. Here, we discuss the applications of marine natural products, in particular, as novel, potent NF-kappaB inhibitors. With the oceans covering two-thirds of the Earth's surface, and with the uniqueness of the environmental properties of marine habitats, it is easily understandable that organisms thriving in the oceans constitute a rich source of chemically unique and biomedically powerful secondary metabolites. Since the early 1960s, significant effort has been placed on the pharmacological evaluation of marine secondary metabolites. Noteworthy achievements of this field of biomedically guided marine exploration, a scientific endeavour often referred to as the search for "Drugs from the Sea", include the discovery of numerous potent anti-cancer, anti-inflammatory, antimicrobial, and analgesic compounds. The chemical characteristics and molecular targets of marine NF-kappaB inhibitors discovered to date are presented and discussed in the context of marine chemical ecology. PMID- 17919456 TI - Induction of G2/M arrest and apoptosis by sesquiterpene lactones in human melanoma cell lines. AB - Malignant melanoma is a highly aggressive tumor which frequently resists chemotherapy, therefore, the search for new agents for its treatment is of great importance. In this study, we purified the sesquiterpene lactones (SLs), Tomentosin and Inuviscolide from Inula viscosa (Compositae) leaves and studied their anti-cancer potency against human melanoma cell lines in order to develop new agents for melanoma treatment. SLs inhibited the proliferation of three human melanoma cell lines: SK-28, 624 mel and 1363 mel in a dose-dependent manner. We further investigated SLs mechanism of action using SK-28 as a representative cell line model. SLs caused cell-cycle arrest at G(2)/M, accompanied by the appearance of a sub-G0 fraction, indicative of apoptotic cell death. Induction of apoptosis was further confirmed by changes in membrane phospholipids, changes in mitochondrial membrane potential (DeltaPsi) and by detection of Caspase-3 activity. Rapid inhibitory phosphorylation of Cdc2 (Thr14 and Tyr15) was seen early after treatment, followed by a later decrease in the expression level of both Cyclin b1 and Cdc2. Induction of p53 and p21(waf1) proteins and phosphorylation of p53 at Ser15 were also detected early after treatment. The anti-apoptotic proteins, p65 subunit of nuclear factor kappaB (NF-kappaB), and Survivin were reduced in a dose-dependent manner. Taken together, these changes partially explain the ability of the SLs to induce G(2)/M arrest and apoptosis. Induction of apoptosis by Tomentosin and Inuviscolide in human aggressive melanoma cell lines has high pharmacological value and implies that SLs might be developed as new agents for melanoma treatment. PMID- 17919457 TI - Effect of bipolar disorder on left frontal cortical responses to goals differing in valence and task difficulty. AB - BACKGROUND: The behavioral activation system (BAS) dysregulation theory of bipolar disorder predicts that bipolar individuals will show an excessive increase in approach motivation during reward striving. Building on past research showing that the left frontal cortical region is involved in approach motivation, we predicted that individuals with bipolar disorder would evidence increased relative left frontal cortical activity in response to goal striving, particularly in response to positive challenges. METHODS: Right-handed individuals (age 18-24) with a bipolar spectrum diagnosis (n = 41) and individuals with no major affective psychopathology (n = 53) were presented with cues indicating that, on a given trial, an easy, medium, or hard anagram (scrambled word) would be presented in 7 seconds and that they would receive money or avoid losing money for the correct solution (10 anagrams of each of the 6 types). During this preparation period, electroencephalogram (EEG) alpha power was measured and hemispheric asymmetry indexes were computed. RESULTS: Compared with the nonbipolar individuals, individuals with bipolar disorder showed greater relative left frontal cortical activation in preparation for the hard/win trials. Whereas nonbipolar individuals showed a decrease in left frontal cortical activation from medium to hard win trials, bipolar individuals did not. In addition, among bipolar individuals, current self-reported activation related to greater left frontal activation to the hard/win trials. CONCLUSIONS: These results provide support for an integrative biopsychosocial model of bipolar disorder, BAS dysregulation theory, and suggest that relative left frontal activity, which may be involved in mania, is triggered by challenging and potentially rewarding events. PMID- 17919458 TI - Mifepristone in psychotic depression? PMID- 17919459 TI - Plasma obestatin concentrations are negatively correlated with body mass index, insulin resistance index, and plasma leptin concentrations in obesity and anorexia nervosa. AB - BACKGROUND: Obestatin is a recently identified ghrelin gene product that was reported to inhibit appetite and gastric motility in contrast to ghrelin. We investigated fasting obestatin and ghrelin levels in patients with obesity and anorexia nervosa. METHODS: Fasting plasma obestatin, acyl-ghrelin, desacyl ghrelin, leptin, glucose serum adiponectin, and insulin were measured in 10 obese subjects, 11 restricting-type anorexics, and 11 control subjects. RESULTS: Obese group had significantly lower levels of obestatin (p < .01), while anorexic group had significantly higher levels (p < .01). Obestatin was negatively correlated with body mass index (BMI) (r = -.74), glucose (r = -.56), insulin (r = -.55), leptin (r = -.66), and also with the homeostasis model assessment of insulin resistance (HOMA-R) (r = -.49) and was positively correlated with acyl-ghrelin (r = .65) and desacyl-ghrelin (r = .60). No correlation was seen between obestatin and adiponectin, but the latter was negatively correlated with both acyl-ghrelin and desacyl-ghrelin. Desacyl-ghrelin to acyl-ghrelin ratio was significantly different between anorexic and control groups (p < .05), while no difference was seen between obese and control groups. CONCLUSIONS: Both obestatin and ghrelin are increased in anorexic and decreased in obesity. We suggest that obestatin is a nutritional marker reflecting body adiposity and insulin resistance. PMID- 17919460 TI - Treatment of SSRI-resistant depression: a meta-analysis comparing within- versus across-class switches. AB - BACKGROUND: Two broad treatment options exist for switching antidepressants for depressed patients who fail to respond to a selective serotonin reuptake inhibitor (SSRI): either a second course of SSRI therapy or a different class of antidepressants. The goal of the present work was to conduct a meta-analysis of studies comparing these two switch strategies. METHODS: Several sources were searched for randomized clinical trials comparing these two switch strategies. RESULTS: Data from four clinical trials (n = 1496) were combined using a random effects model. Patients randomized to switch to a non-SSRI antidepressant (bupropion, mirtazapine, venlafaxine) were more likely to experience remission than patients switched to a second SSRI (risk ratio = 1.29, p = .007). Pooled remission rates were 28% (for non-SSRIs) and 23.5% (for SSRIs). There was also a nonsignificant trend (p = .1) in the rate of discontinuation due to intolerance favoring the within-class switch strategy (risk ratio = 1.23). There was no difference in response rates between the two treatment groups. CONCLUSIONS: These results suggest a modest yet statistically significant advantage in remission rates when switching patients with SSRI-resistant depression to a non-SSRI rather than an SSRI antidepressant. With the number needed to treat (NNT) statistic as one indicator of clinical significance, nearly 22 SSRI nonresponders would need to be switched to a non-SSRI rather than a second SSRI antidepressant to obtain one additional remitter. This difference falls well below the mark of NNT = 10 suggested by the United Kingdom's National Institute of Clinical Excellence but nonetheless might be of public health relevance given the large number of SSRI resistant patients switched to an SSRI versus a non-SSRI antidepressant. PMID- 17919461 TI - DARPP-32 expression in rat brain after electroconvulsive stimulation. AB - Although electroconvulsive therapy (ECT) has been used as a treatment for mental disorder since 1930s, little progress has been made in the mechanisms underlying its therapeutic or adverse effects. The aim of this work was to analyze the expression of DARPP-32 (a protein with a central role in dopaminergic signaling) in striatum, cortex, hippocampus and cerebellum of Wistar rats subjected to acute or chronic electroconvulsive stimulation (ECS). Rats were submitted to a single stimulation (acute) or to a series of eight stimulations, applied one every 48 h (chronic). Animals were killed for collection of tissue samples at time zero, 0.5, 3, 12, 24 and 48 h after stimulation in the acute model and at the same time intervals after the last stimulation in the chronic model. Our results indicated that acute ECS produces smaller changes in the expression of DARPP-32 but, interestingly, chronic ECS increased transient expression of DARPP-32 in several time frames, in striatum and hippocampus, after the last stimulation. Results on the expression of proteins involved in signaling pathways are relevant for neuropsychiatric disorders and treatment, in particular ECT, and can contribute to shed light on the mechanisms related to therapeutic and adverse effects. PMID- 17919462 TI - Evolution of the inflammatory response in the brain following intracerebral hemorrhage and effects of delayed minocycline treatment. AB - There are no effective treatments for intracerebral hemorrhage (ICH). Although inflammation is a potential therapeutic target, there is a dearth of information about time-dependent and cell-specific changes in the expression of inflammation related genes. Using the collagenase-induced ICH model in rats and real-time quantitative RT-PCR we monitored mRNA levels of markers of glial activation, pro- and anti-inflammatory cytokines, enzymes responsible for cytokine activation and several matrix metalloproteases at 6 h and 1, 3 and 7 days after ICH onset. For the most highly up-regulated genes, immunohistochemistry was then used to identify cell-specific protein expression. Finally, minocycline, a drug widely reported to reduce damage in several models of brain injury, was used to test the hypothesis that it can reduce up-regulation of inflammation-related genes when administered using a clinically relevant dosing regime: intraperitoneal injection beginning 6 h after ICH. Our results show a complex inflammatory response, with different brain cell types producing several pro- and anti-inflammatory molecules for at least 7 days after ICH onset. Included is the first demonstration that astrocytes are an important source of interleukin-1beta (IL-1beta), interleukin-1 receptor antagonist (IL-1ra), interleukin-6 (IL-6) and MMP-12. Importantly, our results demonstrate that while delayed minocycline treatment effectively reduces early up-regulation of TNFalpha and MMP-12, its efficacy is lost when treatment is extended for up to a week, and it does not reduce several other genes associated with microglia activation. These results suggest caution in extrapolating to ICH the promising results of minocycline treatment in other models of brain injury. PMID- 17919463 TI - Phase-locking of spontaneous and tone-elicited pontine waves to hippocampal theta waves during REM sleep in rats. AB - Temporal relationships between hippocampal theta waves and pontine waves (P waves) during rapid-eye-movement (REM) sleep were investigated in rats. P waves were phase-locked to the positive theta peak. The phase relationships of P waves elicited by a tone stimulus (P(E) waves) to hippocampal theta waves were also analyzed to qualitatively clarify the mechanism of phase-locking between these two phenomena. P(E) waves occurred at the positive theta peak, as seen for spontaneous P waves. This phase preference of P(E) waves could be understood as that of the response probability to tone stimulus. These data suggest that the P wave generator receives inputs that mimic theta waves. As hippocampal theta waves and P waves are known to be involved in learning and memory processes during REM sleep, the present studies could help to clarify these functions. PMID- 17919465 TI - A new model of reversible superior sagittal sinus thrombosis in rats. AB - To investigate the characteristics of the thrombus at different time points after thrombosis of the intracranial venous sinus, we have developed a new reversible superior sagittal sinus (SSS) model in rats. In this new model, thrombosis was induced by slow injections of the thrombogenic agent into the SSS using a microcatheter. The success of SSS thrombosis was confirmed by magnetic resonance images (MRI), magnetic resonance venographs (MRV), and electron microscopy. T2 weighted MRI and MRV were performed every week for 4 weeks to investigate the process of SSS occlusion. We also examined thrombus formation and the surrounding tissue pathology, as well as endothelial cell injury following SSS occlusion. SSS occlusion occurred at the beginning of the injection of the partial thromboplastin time reagent, and the occluded SSS reopened at the beginning of the second week. MRI images revealed that T2 signals were detected in the parieto occipital lobes 24 h after SSS thrombosis and disappeared at the end of week two. During week two, the rate of thrombus organization was evident and increased significantly in week three. Thrombus calcification was detected in week three and increased significantly in week four. Electron microscopy examination showed the damaged endothelial cell detected at week three following SSS thrombosis. All of these findings suggest that this reversible SSS thrombosis model is feasible and reproducible. The occlusion state can be maintained for at least 4 weeks, providing an opportunity to study the mechanisms of SSS thrombosis. PMID- 17919464 TI - Negative regulation of Vsx1 by its paralog Chx10/Vsx2 is conserved in the vertebrate retina. AB - Chx10/Vsx2 and Vsx1 are the only Paired-like CVC (Prd-L:CVC) homeobox genes in the mouse genome. Both are expressed in the retina and have important but distinct roles in retinal development. Mutations in Chx10/Vsx2 cause reduced retinal progenitor cell (RPC) proliferation and an absence of bipolar cells, while mutations in Vsx1 impair differentiation of cone bipolar cells. Given their structural similarities and importance in retinal development, we sought to determine if a regulatory interaction exists between these genes and whether inactivation of both genes blocks initiation of retinal development. We found that Chx10/Vsx2 binds to a specific sequence in the Vsx1 5'-intergenic region and represses the activity of a luciferase reporter under the control of the Vsx1 promoter. This is consistent with our observation that there is an inverse relationship between the levels of Chx10/Vsx2 and Vsx1 immunostaining within the bipolar cell class. Furthermore, Vsx1 mRNA is upregulated in the RPCs of Chx10/Vsx2 deficient mice and zebrafish embryos injected with a chx10/vsx2 morpholino. In mice deficient for both Chx10/Vsx2 and Vsx1 and zebrafish embryos co-injected with chx10/Vsx2 and vsx1 morpholinos, the changes in embryonic retinal development and marker expression are similar in magnitude to embryos with Chx10/Vsx2 loss of function only. From these studies, we propose that Vsx1 is a direct target of Chx10/Vsx2-mediated transcriptional repression. Although Vsx1 mRNA is upregulated in Chx10/Vsx2 deficient RPCs, Vsx1 does not genetically compensate for loss of Chx10/Vsx2, demonstrating that Prd-L:CVC genes, although important, are not absolutely required to initiate retinal development. PMID- 17919466 TI - Wechsler Memory Scale Revised Edition: neural correlates of the visual paired associates subtest adapted for fMRI. AB - Memory deficits in neurological and psychiatric patients are evaluated by neuropsychological tests such as the Wechsler Memory Scale Revised Edition (WMS R). Neuropsychological data from patients with circumscribed lesions point to single elements of the underlying neural network but fail to identify its whole extent. We report the fMRI adaptation of a subtest of the WMS-R, the Visual Paired Associates. Fifteen healthy, right-handed male volunteers were studied using a 1.5T MRI scanner. The encoding of the combination between a shape and a colour, the assessment of the retrieval of this combination immediately after encoding took place, and the underlying network employed during retrieval a second time after approximately 25 min were investigated. The results show a fronto-parieto-occipital network with left frontal accentuation for encoding and a fronto-parieto-occipital network for immediate and delayed retrieval. Noteworthy is the specific role of the thalamus. During immediate retrieval, the thalamus showed significant bilateral activation; during delayed retrieval, there was no significant activation. The thalami are part of an extended hippocampal diencephalic system which is critical for efficient encoding and normal retrieval of new episodic information. We describe the probability of thalamocortical connections during retrieval based on the Thalamus Connectivity Atlas. The cerebellum showed significant activation in all conditions; its part in higher cognitive functions such as memory was thereby confirmed. PMID- 17919468 TI - Effects of LiCl/pilocarpine-induced status epilepticus on rat brain mu and benzodiazepine receptor binding: regional and ontogenetic studies. AB - Neurochemical studies document involvement of benzodiazepine (BDZ) and mu opioid receptors in seizure development and their possible age-related role during epileptogenesis. To study developmental changes of this role LiCl/pilocarpine status epilepticus (SE) was induced in P12, P25 and/or adult rats. This SE leads to epilepsy in all adult and subpopulation of immature rats. Using in vitro autoradiography, benzodiazepine (BDZ) and mu opioid receptor binding was evaluated 1 week (early phase of epileptogenesis) and 3 months (chronic phase) after SE in 27 brain structures involved in seizure generation and spread (amygdala, hippocampus, basal ganglia and thalamic nuclei). The pattern of receptor binding changes was related to the age at SE, interval after SE and to brain structures. Enhanced BDZ binding was found 1 week after SE in many cortical areas in P12 and also in the amygdala complex and dentate gyrus in both P12 and P25. No changes of BDZ binding occurred in adults at that time, but 3 months after SE a decrease of binding appeared in all evaluated areas in both adult and P25 but not P12 rats. This decrease did not reflect neuronal loss. mu opioid receptors were less significantly affected but clear tendency to decrease binding occurred in adult rats in various cortical, amygdala and thalamic regions early after SE. Changes were less expressed in immature rats. Our data support the hypothesis that age-related changes of receptor properties may participate in different functional consequences of SE including epileptogenesis (more common in older age groups) and behavioral changes. PMID- 17919467 TI - A prototypical Sigma-1 receptor antagonist protects against brain ischemia. AB - Previous studies indicate that the Sigma-1 ligand 4-phenyl-1-(4-phenylbutyl) piperidine (PPBP) protects the brain from ischemia. Less clear is whether protection is mediated by agonism or antagonism of the Sigma-1 receptor, and whether drugs already in use for other indications and that interact with the Sigma-1 receptor might also prevent oxidative damage due to conditions such as cerebral ischemic stroke. The antipsychotic drug haloperidol is an antagonist of Sigma-1 receptors and in this study it potently protects against oxidative stress related cell death in vitro at low concentrations. The protective potency of haloperidol and a number of other butyrophenone compounds positively correlate with their affinity for a cloned Sigma-1 receptor, and the protection is mimicked by a Sigma-1 receptor-selective antagonist (BD1063), but not an agonist (PRE 084). In vivo, an acute low dose (0.05 mg/kg s.c.) of haloperidol reduces by half the ischemic lesion volume induced by a transient middle cerebral artery occlusion. These in vitro and in vivo pre-clinical results suggest that a low dose of acutely administered haloperidol might have a novel application as a protective agent against ischemic cerebral stroke and other types of brain injury with an ischemic component. PMID- 17919469 TI - Changes in the rates of the tricarboxylic acid (TCA) cycle and glutamine synthesis in the monkey brain with hemiparkinsonism induced by intracarotid infusion of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP): studies by non invasive 13C-magnetic resonance spectroscopy. AB - The tricarboxylic acid cycle rate (Vtca) and the rate of glutamine synthesis (Vgln) in the pre- and post-MPTP-treated cynomolgus monkey (Macaca fascicularis) brain were measured non-invasively using a 2 Tesla 13C-magnetic resonance spectroscopy (13C-MRS; multislice 1H-13C correlation heteronuclear single quantum coherence spectroscopy) system. Before the infusion of 1-methyl-4-phenyl-1,2,3,6 tertahydropyridine (MPTP) into three monkeys, spectra were obtained by 13C-MRS from each monkey under anesthesia after the bolus injection of [1-13C] glucose (99% atom excess, 0.28 g/kg) followed by the continuous infusion of [1-13C] glucose (99% atom excess, 0.72 g/kg) into the saphenous vein for 3 h. The average values of Vtca were 0.475+/-0.077 (mean+/-S.D.) and 0.472+/-0.073 micromol/g/min, and the average values of Vgln were 0.042+/-0.007 and 0.041+/-0.008 mumol/g/min on the left and on the right hemisphere, respectively. Three monkeys were induced hemiparkinsonism by intracarotid (left) infusion of MPTP (0.6 mg/kg) and then were employed in 13C-MRS studies for 2 (5, 14 days), 3 (3, 8, 71 days) or 4 (5, 11, 27, 78 days) times, respectively, after the MPTP treatment. The average ratios of Vtca and Vgln on the left hemisphere to those on the right hemisphere in pre- and post-MPTP-treated monkeys were 0.837+/-0.085 and 1.373+/-0.132, respectively. These results of non-invasive 13C-MRS analysis of the MPTP primate model of Parkinson's disease indicate that the loss of the dopaminergic innervation from the caudate putamen may modulate the overall glucose metabolism to glutamate and glutamine in the ipsilateral cerebrum. PMID- 17919470 TI - Pupil dilatation in response to vagal afferent electrical stimulation is mediated by inhibition of parasympathetic outflow in the rat. AB - Pupil dilatation (PD) was produced by vaginocervical mechanical stimulation (VCMS) in intact rats, and in rats with spinal cord (SC) transection at level T7 (i.e., above the level of entry into SC of all genitospinal nerves). After transection of SC, bilateral vagotomy abolished PD in response to VCMS, providing evidence that the vagus nerve conveys VCMS directly from the genitalia to the brain. In the present study, unilateral electrical stimulation of the central stump of the vagus nerve (ESV) transected at the spinal cervical level produced PD in otherwise intact rats, and in rats that had undergone bilateral cervical sympathectomy (CSX). Unilateral ESV produced bilaterally symmetrical PD. By contrast, unilateral electrical stimulation of the cervical sympathetic chain produced PD only ipsilateral to stimulation. These results provide evidence that activation of sympathetic outflow, although sufficient to produce PD, does not, by itself, mediate PD in response to ESV. After bilateral CSX, transection of the brain stem at the mid-pontine level abolished PD in response to unilateral ESV, while preserving the pupillary light reflex. Since CSX eliminates all sympathetic control of the pupil, and transection of the brain stem at the mid-pontine level blocks access of vagus-induced activity that ascends to the oculomotor nucleus in the midbrain, we conclude that ESV produces PD, at least in part, via inhibition of output from the (parasympathetic) oculomotor nucleus. PMID- 17919471 TI - Contribution of the GABAergic pathway(s) to the correlated activities of chicken retinal ganglion cells. AB - In the present study, the spatiotemporal pattern of chicken retinal ganglion cells' firing activity in response to full-field white light stimulation was investigated. Cross-correlation analysis showed that ganglion cells of sustained subtype fired in precise synchrony with their adjacent neurons of the same subtype (delay lag within 2 ms, narrow correlation). On the other hand, the activities of neighboring ganglion cells of transient subtype were correlated with distributed time lags (10-30 ms, medium correlation). Pharmacological studies demonstrated that the intensity of the medium correlations could be strengthened when exogenous GABA was applied and attenuated when GABA receptors were blocked by picrotoxin. Meanwhile, the GABAergic modulation on the narrow correlations was not consistent. These results suggest that, in the chicken retina, GABAergic pathway(s) are likely involved in the formation of medium correlations between ganglion cells. Neurons might fire at a lower rate but with higher level of synchronization to improve the efficiency of information transmission, with the mechanism involving the GABAergic inhibitory input. PMID- 17919472 TI - Working with what we have: options for myocardial infarct repair? PMID- 17919479 TI - Best hospitals? PMID- 17919480 TI - Nuclear hormone receptors and intestinal inflammation. PMID- 17919481 TI - Recent advances in assessing anorectal structure and functions. PMID- 17919482 TI - Clinical challenges and images in GI. Gallstone impacted in duodenum causing gastric outlet obstruction (Bouveret syndrome). PMID- 17919483 TI - Clinical Challenges and Images in GI. Unique gastrointestinal involvement in a patient with Henoch-Schonlein purpura. PMID- 17919484 TI - Colonoscopic screening of first-degree relatives of patients with large adenomas: increased risk of colorectal tumors. AB - BACKGROUND & AIMS: The risk of developing colorectal neoplasia is not well established among family members of individuals with large adenomas, and screening strategies remain under debate in this population. This study aimed at quantifying the risk of colorectal adenomas and cancers using colonoscopic screening in first-degree relatives of patients with large adenomas. METHODS: This case-control study was performed in 18 endoscopic units of French nonuniversity hospitals. A colonoscopy was offered to first-degree relatives of 306 index cases with adenomas > or =10 mm if they were alive, aged 40-75 years, and could be contacted by the index case. Among them, 168 were examined and matched for age, sex, and geographical area with 2 controls (n = 307). Controls were randomly selected from 1362 consecutive patients aged 40-75 years having undergone a colonoscopy for minor symptoms. RESULTS: The prevalence of large adenomas and cancers was 8.4% and 4.2%, in relatives and controls, respectively. Odds ratios (ORs) associated with a history of large adenomas in relatives were 2.27 (95% confidence interval [CI], 1.01-5.09) for cancers or large adenomas, 1.21 (95% CI, 0.68-2.15) for small adenomas, and 1.56 (95% CI, 0.96-2.53) for all colorectal neoplasia. The risk of large adenomas and cancers was higher in relatives of index cases younger than 60 years (OR, 3.82; 95% CI, 0.92-15.87) and when the index case had large distal adenomas (OR, 3.14; 95% CI, 1.27-7.73). CONCLUSIONS: First-degree relatives of patients with large adenomas are at increased risk of developing colorectal cancers or large adenomas. This result has implications for screening in this high-risk population. PMID- 17919485 TI - Development of colorectal tumors in colonoscopic surveillance in Lynch syndrome. AB - BACKGROUND & AIMS: Mutation carriers in Lynch syndrome families have a high risk for developing colorectal cancer during their lifetime. This study was designed to assess the cumulative risk for the development of colorectal adenoma or carcinoma in prospective colonoscopic surveillance. METHODS: Data from the Finnish Hereditary Colorectal Cancer Registry electronic database on 420 Lynch syndrome mutation carriers without previous colorectal tumors were reviewed. Between March 1982 and May 2005 the mutation carriers underwent a total of 1252 colonoscopies. The total follow-up time was 3150 years (mean, 6.7 y/patient). RESULTS: The cumulative risk of adenoma by age 60 was estimated as 68% (95% confidence interval [CI], 50%-80%) in men and 48% (95% CI, 29%-62%) in women. The estimated cumulative risk up to age 60 years for the development of cancer found as a result of surveillance at an interval of 2-3 years was 35% (95% CI, 16%-49%) in men and 22% (95% CI, 7%-34%) in women. Half of the adenomas were located proximal to the splenic flexure. Extracolonic cancer was diagnosed in 73 patients (18%). CONCLUSIONS: Adenoma would appear to be the most important lesion preceding cancer formation in Lynch syndrome and removal of adenomas decreases the risk for colorectal cancer (CRC). The Finnish surveillance protocol of colonoscopies at 2- to 3-year intervals facilitates patient adherence but includes an essential risk for CRC up to 60 years of age, but without CRC-related mortality when the surveillance instructions are followed. PMID- 17919486 TI - Histologic inflammation is a risk factor for progression to colorectal neoplasia in ulcerative colitis: a cohort study. AB - BACKGROUND & AIMS: Although inflammation is presumed to contribute to colonic neoplasia in ulcerative colitis (UC), few studies have directly examined this relationship. Our aim was to determine whether severity of microscopic inflammation over time is an independent risk factor for neoplastic progression in UC. METHODS: A cohort of patients with UC undergoing regular endoscopic surveillance for dysplasia was studied. Degree of inflammation at each biopsy site had been graded as part of routine clinical care using a highly reproducible histologic activity index. Progression to neoplasia was analyzed in proportional hazards models with inflammation summarized in 3 different ways and each included as a time-changing covariate: (1) mean inflammatory score (IS-mean), (2) binary inflammatory score (IS-bin), and (3) maximum inflammatory score (IS-max). Potential confounders were analyzed in univariate testing and, when significant, in a multivariable model. RESULTS: Of 418 patients who met inclusion criteria, 15 progressed to advanced neoplasia (high-grade dysplasia or colorectal cancer), and 65 progressed to any neoplasia (low-grade dysplasia, high-grade dysplasia, or colorectal cancer). Univariate analysis demonstrated significant relationships between histologic inflammation over time and progression to advanced neoplasia (hazard ration (HR), 3.0; 95% CI: 1.4-6.3 for IS-mean; HR, 3.4; 95% CI: 1.1-10.4 for IS-bin; and HR, 2.2; 95% CI: 1.2-4.2 for IS-max). This association was maintained in multivariable proportional hazards analysis. CONCLUSIONS: The severity of microscopic inflammation over time is an independent risk factor for developing advanced colorectal neoplasia among patients with long-standing UC. PMID- 17919487 TI - Altered rectal perception in irritable bowel syndrome is associated with symptom severity. AB - BACKGROUND & AIMS: Diverging results exist regarding the connection between altered visceral perception and gastrointestinal (GI) symptoms, as well as the effects of psychological status on visceral sensitivity. We sought to investigate different aspects of rectal perception in irritable bowel syndrome (IBS) and the association with GI and psychological symptoms. METHODS: We included 109 patients with IBS meeting Rome II criteria (77 women; age range, 20-71 years) and 29 healthy controls (21 women; age range, 20-68 years). They underwent rectal balloon distentions determining sensory thresholds for discomfort and pain, the perceived intensity of unpleasantness, and the viscerosomatic referral area. The fifth percentile (thresholds) and 95th percentile (unpleasantness and referral area) in controls were used to define altered perception. Questionnaires were used to assess severity of IBS-related GI symptoms and psychological symptoms. RESULTS: When combining the 3 aspects of perception, 67 patients (61%) had altered rectal perception. These patients, compared with normosensitive patients, more frequently reported moderate or severe pain (73% vs 44%; P < .01), bloating (73% vs 36%; P < .0001), diarrhea (47% vs 21%; P < .01), satiety (39% vs 13%; P < .01), and clinically significant anxiety (31% vs 12%; P < .05). In a multivariate analysis, only pain and bloating remained associated with altered rectal perception. CONCLUSIONS: Altered rectal perception is common in IBS and seems to be one important pathophysiologic factor associated with GI symptom severity in general and pain and bloating in particular. It is not just a reflection of the psychological state of the patient. PMID- 17919488 TI - Free fatty acids have more potent effects on gastric emptying, gut hormones, and appetite than triacylglycerides. AB - BACKGROUND & AIMS: The effects of fat on gastric emptying (GE), gut hormones, and energy intake are dependent on digestion to free fatty acids (FFAs). In animals, small intestinal oleic acid inhibits energy intake more potently than the triacylglyceride (TG) triolein, but there is limited information about the comparative effects of FFA and TG in human beings. We compared the effects of FFA and TG on GE, gut hormone secretion, appetite, and energy intake in healthy males. METHODS: Nine men (age, 23 +/- 2 y; body mass index, 22 +/- 1 kg/m(2)) were studied on 3 occasions to evaluate the effects of (1) 40 g oleic acid (FFA, 1830 kJ), (2) 40 g macadamia oil (TG, 1856 kJ; both 600-mL oil-in-water emulsions stabilized with 4% milk protein and labeled with 15 MBq (123)I), or (3) 600 mL 4% milk protein (control, 352 kJ), administered intragastrically, on GE, plasma cholecystokinin (CCK) and peptide-YY (PYY) levels, appetite perceptions, and subsequent energy intake. RESULTS: GE of FFA was much slower than that of TG (P < .05), with greater retention of FFA, than TG, in the proximal stomach (P < .001). Hunger was less (P < .05), and fullness was greater (P < .05), after FFA when compared with control and TG. Increases in plasma CCK and PYY levels were greater after FFA than TG or control (P < .05). Energy intake tended to be less after FFA compared with TG (control, 4754 +/- 610 kJ; TG, 5463 +/- 662 kJ; FFA, 4199 +/- 410 kJ). CONCLUSIONS: FFAs empty from the stomach more slowly, but stimulate CCK and PYY and suppress appetite more potently than TG in healthy human beings. PMID- 17919489 TI - HCV-specific T-cell response in relation to viral kinetics and treatment outcome (DITTO-HCV project). AB - BACKGROUND & AIMS: The second slope of viral decline induced by interferon treatment has been suggested to be influenced mainly by the hepatitis C virus (HCV)-specific T-cell response; however, this hypothesis needs to be validated by results derived from experimental studies. METHODS: To address this issue, the HCV-specific T-cell response of 32 genotype-1-infected patients of the 270 patients enrolled in the dynamically individualized treatment of hepatitis C infection and correlates of viral/host dynamics phase III, open-label, randomized, multicenter trial was studied in relation to viral kinetics and treatment outcome. RESULTS: Greater proliferative responses by HCV-specific CD8 cells were found before treatment in patients with a fast viral decline and with a sustained viral response. However, no significant improvement of HCV-specific CD8 responses was observed in the first weeks of therapy in both rapid viral responder and non-rapid viral responder patients. A mild enhancement of proliferative T-cell responses and a partial restoration of the cytotoxic T-cell potential was expressed only late during treatment, likely favored by HCV clearance. CONCLUSIONS: Early restoration of an efficient T-cell response does not seem to be an essential requirement for a rapid viral decline in the first weeks of treatment. However, patients presenting a better HCV-specific CD8 cell proliferative potential at baseline are more likely to present a rapid and sustained viral response. Therefore, future treatment protocols should consider the development of strategies aimed at improving HCV-specific T-cell responses. PMID- 17919490 TI - Ultra-rapid cardiotoxicity of the hepatitis C virus protease inhibitor BILN 2061 in the urokinase-type plasminogen activator mouse. AB - BACKGROUND & AIMS: Because current therapies for chronic hepatitis C virus (HCV) infections are suboptimal and associated with severe side effects, novel treatment options are needed. A small animal model has recently been developed to study HCV infections. To examine the usefulness of this human liver-urokinase type plasminogen activator (uPA)(+/+) severe combined immune deficient (SCID) mouse for the development of HCV-targeted drugs, we evaluated the antiviral efficacy and safety of an HCV NS3-protease inhibitor, BILN 2061. METHODS: BILN 2061 was orally administered at clinical range doses for 4 days to SCID mice that differed in the presence of HCV infection, human hepatocyte grafts, and uPA zygosity. Treatment outcome was evaluated clinically, virologically, and morphologically. Using standard high-performance liquid chromatography ultraviolet (HPLC-UV) methods and mass spectrometry, single-dose pharmacokinetics and multiple-dose drug exposures were analyzed. The (13)C-aminopyrine breath test was applied to compare in vivo liver function. RESULTS: A 4-day treatment with BILN 2061 of HCV genotype-1b infected chimeric animals reduced the viral load by >100-fold, but concomitant clinical and ultrastructural signs of cardiotoxicity appeared. BILN 2061 administration to uPA-transgenic mice induced mitochondrial swelling with aberrant cristae in cardiomyocytes, but not in skeletal muscle. Because both drug accumulation and liver function were identical in affected uPA transgenic and nontransgenic SCID mice without cardiac involvement, the urokinase plasminogen activator transgene itself appears to be implicated. CONCLUSIONS: The human liver-uPA(+/+)SCID mouse is an interesting small animal model to evaluate the preclinical safety and efficacy of new antiviral compounds against HCV. The uPA-transgene increases the susceptibility of mice to BILN 2061-induced cardiotoxicity. PMID- 17919491 TI - Antiviral activity and hepatoprotection by heme oxygenase-1 in hepatitis B virus infection. AB - BACKGROUND & AIMS: Induction of heme oxygenase-1 (HO-1) has been shown to be beneficial in immune-mediated liver damage. We now investigate the effects of HO 1 induction in models of human hepatitis B virus (HBV) infection. METHODS: Adenoviral transfer of an HBV 1.3 genome into wild-type mice was used as a model for acute hepatitis B. HBV transgenic animals were used as a model for chronic HBV infection. HBV replication was assessed by HBV viremia, antigenemia, and Southern blotting, liver damage was assessed by serum alanine aminotransferase activities and histopathology of liver sections. To investigate HO-1 effects on HBV replication at a molecular level, stably HBV-transfected hepatoma cells were used. HBV gene expression, protein stability, transcription, and replication were determined. HO-1 was induced by either cobalt-protoporphyrin-IX or over expressed by adenoviral gene transfer. RESULTS: In the acute hepatitis B model, liver injury was reduced significantly after HO-1 induction. In addition, HO-1 showed a pronounced antiviral effect, which was confirmed in stably HBV-transfected hepatoma cells and in persistently HBV replicating transgenic mice. We showed that HO-1 induction repressed HBV replication directly in hepatocytes at a posttranscriptional step by reducing stability of HBV core protein and thus blocking refill of nuclear HBV covalently closed circular (ccc)DNA. Small interfering RNA directed against HO-1 proved that this effect depended on the expression level of HO-1. CONCLUSIONS: Besides its hepatoprotective effect, HO-1 showed a pronounced antiviral activity in HBV infection. Therefore, induction of HO-1 might be a novel therapeutic option for inflammatory flares of hepatitis B. PMID- 17919493 TI - Evidence for the role of interferon-alfa production by dendritic cells in the Th1 response in celiac disease. AB - BACKGROUND & AIMS: Dendritic cells (DCs) play a crucial role in immune responses by controlling the extent and type of T-cell response to antigen. Celiac disease is a condition in which T-cell immunity to gluten plays an important pathogenic role, yet information on DCs is scant. We examined mucosal DCs in celiac disease in terms of phenotype, activation/maturation state, cytokine production, and function. METHODS: Mucosal DCs from 48 celiacs and 30 controls were investigated by flow cytometry. In situ distribution of DCs was analyzed by confocal microscopy. Interferon (IFN)-alfa, interleukin (IL)-4, IL-5, IL-12p35, IL-12p40, IL-18, IL-23p19, IL-27, and transforming growth factor-beta transcripts were measured by real-time reverse-transcription polymerase chain reaction in sorted DCs. DC expression of IL-6, IL-12p40, and IL-10 was assessed by intracellular cytokine staining. The effect of IFN-alfa and IL-18 blockade on the gluten induced IFN-gamma response in celiac biopsy specimens grown ex vivo also was investigated. RESULTS: Mucosal DCs were increased in untreated, but not treated, celiacs. The majority of them were plasmacytoid with higher levels of maturation (CD83) and activation (CD80/CD86) markers. Higher transcripts of Th1 relevant cytokines, such as IFN-alfa, IL-18, and IL-23p19, were produced by celiac DCs, but because IL-12p40 was undetectable, a role for IL-23 is unlikely. Intracellular cytokine staining of celiac DCs showed higher IL-6, but lower IL-10 expression, and confirmed the lack of IL-12p40. Blocking IFN-alfa inhibited IFN gamma transcripts in ex vivo organ culture of celiac biopsy specimens challenged with gluten. CONCLUSIONS: These data suggest that IFN-alfa-producing DCs contribute to the Th1 response in celiac disease. PMID- 17919494 TI - Nitric oxide and acid induce double-strand DNA breaks in Barrett's esophagus carcinogenesis via distinct mechanisms. AB - BACKGROUND & AIMS: The luminal microenvironment including acid and nitric oxide (NO) has been implicated in Barrett's esophagus carcinogenesis. We investigated the ability of acid and NO to induce DNA damage in esophageal cells. METHODS: Transformed and primary Barrett's esophagus and adenocarcinoma cells were exposed to either acid, (pH 3.5), +/- antioxidant or NO from a donor or generated by acidification of nitrite in the presence of ascorbate +/- NO scavenger. Phosphorylation of histone H2AX and the neutral comet assay were used to detect DNA double-strand breaks (DSBs). Intracellular levels of reactive oxygen species and NO were detected with fluorescent dyes. Mitochondrial viability was measured with a rhodamine dye. Long-term survival was assessed by clonogenic assay. RESULTS: Exposure to acid (pH 3.5) for > or =15 minutes induced DSBs in all cell lines (P < .05). There was a concomitant increase in intracellular reactive oxygen species in the absence of mitochondrial damage, and pretreatment with antioxidants inhibited DNA damage. Exposure to physiologic concentrations of NO produced from the NO donor or acidification of salivary nitrite induced DSBs in a dose- (>25 micromol/L) and cell-dependent manner (adenocarcinoma >Barrett's esophagus, P < .05). This occurred preferentially in S-phase cells consistent with stalled replication forks and was blocked with a NO scavenger. NO also induced DSBs in primary Barrett's esophagus cells treated ex vivo. Cells were able to survive when exposed to acid and NO. CONCLUSIONS: Both acid and NO have the potential to generate DSBs in the esophagus and via distinct mechanisms. PMID- 17919492 TI - Reciprocal effects of micro-RNA-122 on expression of heme oxygenase-1 and hepatitis C virus genes in human hepatocytes. AB - BACKGROUND & AIMS: Heme oxygenase-1 (HO-1) is an antioxidant defense and key cytoprotective enzyme, which is repressed by Bach1. Micro-RNA-122 (miR-122) is specifically expressed and highly abundant in human liver and required for replication of hepatitis C virus (HCV) RNA. This study was to assess whether a specific miR-122 antagomir down-regulates HCV protein replication and up regulates HO-1. METHODS: We transfected antagomir of miR-122, 2'-O-methyl-mimic miR-122, or nonspecific control antagomir, into wild-type (WT) Huh-7 cells or Huh 7 stably replicating HCV subgenomic protein core through nonstructural protein 3 of HCV (NS3) (CNS3 replicon cells) or NS3-5B (9-13 replicon cells). RESULTS: Antagomir of miR-122 reduced the abundance of HCV RNA by 64% in CNS3 and by 84% in 9-13 cells. Transfection with 2'-O-methlyl-mimic miR-122 increased HCV levels up to 2.5-fold. Antagomir of miR-122 also decreased Bach1 and increased HO-1 mRNA levels in CNS3, 9-13, and WT Huh-7 cells. Increasing HO-1 by silencing Bach1 with 50 nmol/L Bach1-short interfering RNA or by treatment with 5 mumol/L cobalt protoporphyrin or heme (known inducers of HO-1) decreased HCV RNA and protein by 50% in HCV replicon cells. CONCLUSIONS: Down-regulation of HCV replication using an antagomir targeted to miR-122 is effective, specific, and selective. Increasing HO-1, by silencing the Bach1 gene or by treatment with cobalt protoporphyrin or heme, decreases HCV replication. Thus, miR-122 plays an important role in the regulation of HCV replication and HO-1/Bach1 expression in hepatocytes. Down-regulation of miR-122 and up-regulation of HO-1 may be new strategies for anti-HCV intervention and cytoprotection. PMID- 17919495 TI - Muc1 mucin limits both Helicobacter pylori colonization of the murine gastric mucosa and associated gastritis. AB - BACKGROUND & AIMS: The MUC1 mucin is expressed on the cell surface of epithelial cells lining the gastric mucosa. Epidemiologic studies suggest that functional allelic variations in the MUC1 gene may play a role in human susceptibility to Helicobacter pylori-associated pathologies, including gastric adenocarcinoma. We have evaluated the impact of Muc1 expression on the colonization and pathogenesis of gastric Helicobacter infections. METHODS: Wild-type and Muc1-deficient mice were infected with H pylori and colonization and gastritis levels determined. Primary gastric cells were used to examine the impact of Muc1 expression on bacterial adherence. RESULTS: Mice lacking Muc1 were colonized by 5-fold more H pylori within 1 day of infection, and this difference was maintained for at least 2 months postinfection. Mice heterozygous for the null Muc1 allele developed intermediate bacterial colonization. Although wild-type mice developed only a mild gastritis when infected for 2 months with H pylori, Muc1(-/-) mice developed an atrophic gastritis marked by loss of parietal cells. We demonstrate H pylori adhesion to purified MUC1 and significantly increased adhesion to cultured murine Muc1 null gastric epithelial cells, suggesting that Muc1 acts as a decoy limiting binding to the cell surface. CONCLUSIONS: Muc1 provides a protective barrier, which limits both acute and chronic colonization by H pylori, as well as playing a major role in limiting the inflammation induced by Helicobacter infection. We propose that Muc1 restricts access of H pylori to the epithelial surface, hence reducing exposure of the host to proinflammatory bacterial products. PMID- 17919496 TI - Activation of the cholinergic anti-inflammatory pathway ameliorates postoperative ileus in mice. AB - BACKGROUND & AIMS: We previously showed that intestinal inflammation is reduced by electrical stimulation of the efferent vagus nerve, which prevents postoperative ileus in mice. We propose that this cholinergic anti-inflammatory pathway is mediated via alpha7 nicotinic acetylcholine receptors expressed on macrophages. The aim of this study was to evaluate pharmacologic activation of the cholinergic anti-inflammatory pathway in a mouse model for postoperative ileus using the alpha7 nicotinic acetylcholine receptor-agonist AR-R17779. METHODS: Mice were pretreated with vehicle, nicotine, or AR-R17779 20 minutes before a laparotomy (L) or intestinal manipulation (IM). Twenty-four hours thereafter gastric emptying was determined using scintigraphy and intestinal muscle inflammation was quantified. Nuclear factor-kappaB transcriptional activity and cytokine production was assayed in peritoneal macrophages. RESULTS: Twenty-four hours after surgery IM led to a delayed gastric emptying compared with L (gastric retention: L(saline) 14% +/- 4% vs IM(saline) 38% +/- 10%, P = .04). Pretreatment with AR-R17779 prevented delayed gastric emptying (IM(AR R17779) 15% +/- 4%, P = .03). IM elicited inflammatory cell recruitment (L(saline) 50 +/- 8 vs IM(saline) 434 +/- 71 cells/mm(2), P = .001) which was reduced by AR-R17779 pretreatment (IM(AR-R17779) 231 +/- 32 cells/mm(2), P = .04). An equimolar dose of nicotine was not tolerated. Subdiaphragmal vagotomy did not affect the anti-inflammatory properties of AR-R17779. In peritoneal macrophages, both nicotinic agonists reduced nuclear factor kappaB transcriptional activity and proinflammatory cytokine production, with nicotine being more effective than AR-R17779. CONCLUSIONS: AR-R17779 treatment potently prevents postoperative ileus, whereas toxicity limits nicotine administration to ineffective doses. Our data further imply that nicotinic inhibition of macrophage activation may involve other receptors in addition to alpha7 nicotinic acetylcholine receptor. PMID- 17919497 TI - Clock gene expression in the murine gastrointestinal tract: endogenous rhythmicity and effects of a feeding regimen. AB - BACKGROUND & AIMS: Based on observations that the gastrointestinal tract is subject to various 24-hour rhythmic processes, it is conceivable that some of these rhythms are under circadian clock gene control. We hypothesized that clock genes are present in the gastrointestinal tract and that they are part of a functional molecular clock that coordinates rhythmic physiologic functions. METHODS: The effects of timed feeding and vagotomy on temporal clock gene expression (clock, bmal1, per1-3, cry1-2) in the gastrointestinal tract and suprachiasmatic nucleus (bmal, per2) of C57BL/6J mice were examined using real time polymerase chain reaction and Western blotting (BMAL, PER2). Colonic clock gene localization was examined using immunohistochemistry (BMAL, PER1-2). RESULTS: Clock immunoreactivity was observed in the myenteric plexus and epithelial crypt cells. Clock genes were expressed rhythmically throughout the gastrointestinal tract. Timed feeding shifted clock gene expression at the RNA and protein level but did not shift clock gene expression in the central clock. Vagotomy did not alter gastric clock gene expression compared with sham-treated controls. CONCLUSIONS: The murine gastrointestinal tract contains functional clock genes, which are molecular core components of the circadian clock. Daytime feeding in nocturnal rodents is a strong synchronizer of gastrointestinal clock genes. This synchronization occurs independently of the central clock. Gastric clock gene expression is not mediated through the vagal nerve. The presence of clock genes in the myenteric plexus and epithelial cells suggests a role for clock genes in circadian coordination of gastrointestinal functions such as motility, cell proliferation, and migration. PMID- 17919498 TI - Lack of haptoglobin affects iron transport across duodenum by modulating ferroportin expression. AB - BACKGROUND & AIMS: Haptoglobin is an acute phase protein responsible for the recovery of free hemoglobin from plasma. Haptoglobin-null mice were previously shown to have an altered heme-iron distribution, thus reproducing what occurs in humans in cases of congenital or acquired anhaptoglobinemia. Here, we report the analysis of iron homeostasis in haptoglobin-null mice. METHODS: Iron absorption was measured in tied-off duodenal segments. Iron stores were evaluated on tissue homogenates and sections. The expression of molecules involved in iron homeostasis was analyzed at the protein and messenger RNA levels both in mice and in murine RAW264.7 macrophages stimulated in vitro with hemoglobin. RESULTS: Analysis of intestinal iron transport reveals that haptoglobin-null mice export significantly more iron from the duodenal mucosa to plasma compared with control counterparts. Increased iron export from the duodenum correlates with increased duodenal expression of ferroportin, both at the protein and messenger RNA levels, whereas hepatic hepcidin expression remains unchanged. Up-regulation of the ferroportin transcript, but not of the protein, also occurs in haptoglobin-null spleen macrophages, which accumulate free hemoglobin-derived iron. Finally, we demonstrate that hemoglobin induces ferroportin expression in RAW264.7 cells. CONCLUSIONS: Taking together these data, we suggest that haptoglobin, by controlling plasma levels of hemoglobin, participates in the regulation of ferroportin expression, thus contributing to the regulation of iron transfer from duodenal mucosa to plasma. PMID- 17919499 TI - The effect of statins in colorectal cancer is mediated through the bone morphogenetic protein pathway. AB - BACKGROUND & AIMS: Epidemiological evidence suggests that statins prevent colorectal cancer (CRC), but the biological mechanism remains obscure. Statins induce bone morphogenetic protein (BMP) expression in bone cells. We have previously shown that BMPs act as tumor suppressors in CRC. We hypothesized that the action of statins in CRC involves the induction of BMPs. METHODS: We investigated the effects of statins on CRC cell lines using immunoblotting, measurements of apoptosis and cell proliferation, and luciferase reporter assays. The effect of statins was confirmed in a xenograft mouse model. RESULTS: CRC cell lines show widely differing sensitivities to statin treatment. Sensitive cell lines show induction of BMP2 protein levels and a BMP2 reporter construct, activation of the BMP pathway, and induction of the BMP target gene ID-2, whereas resistant cell lines do not. The addition of the specific inhibitor of BMPs, noggin, completely prevents lovastatin-induced apoptosis in sensitive cells. Sensitive cell lines express the central BMP pathway element SMAD4, whereas the resistant cell lines do not. Targeted knockout of SMAD4 leads to the loss of statin sensitivity and reconstitution with SMAD4, to the restoration of statin sensitivity. In a xenograft mouse model, tumors from sensitive and insensitive cell lines were treated with oral simvastatin. Significant inhibition of tumor growth using sensitive cells but increased tumor growth when using insensitive cells was observed. CONCLUSIONS: Statins induce apoptosis in CRC cells through induction of BMP2. Statin therapy may only be effective in SMAD4-expressing CRCs and may have adverse effects in SMAD4-negative tumors. PMID- 17919500 TI - Bacterial endotoxin: a trigger factor for alcoholic pancreatitis? Evidence from a novel, physiologically relevant animal model. AB - BACKGROUND & AIMS: This study examined the possible role of endotoxinemia (from increased gut permeability) as an additional trigger factor for overt pancreatic disease and as a promoter of chronic pancreatic injury in alcoholics by using a rat model of chronic alcohol feeding and in vitro experiments with cultured pancreatic stellate cells (PSCs), the key mediators of pancreatic fibrosis. METHODS: In the in vivo model, Sprague-Dawley rats fed isocaloric Lieber-DeCarli liquid diets +/- alcohol for 10 weeks were challenged with a single dose or 3 repeated doses of the endotoxin lipopolysaccharide (LPS) and the pancreas was examined. In the in vitro studies, rat PSCs were assessed for activation on exposure to LPS +/- ethanol. The expression of LPS receptors TLR4 and CD14 also was assessed in rat and human PSCs. RESULTS: In the in vivo model, single or repeated LPS challenge resulted in significantly greater pancreatic injury in alcohol-fed rats compared with rats fed the control diet without alcohol. Notably, repeated LPS injections caused pancreatic fibrosis in alcohol-fed rats, but not in rats fed the control diet. In the in vitro studies, PSCs were activated by LPS. Alcohol + LPS exerted a synergistic effect on PSC activation. Importantly, both rat and human PSCs expressed TLR4 and CD14. CONCLUSIONS: This study describes, for the first time, a clinically relevant animal model of alcohol-related pancreatic injury and provides strong in vivo and in vitro evidence that suggests that LPS is a trigger factor in the initiation and progression of alcoholic pancreatitis. PMID- 17919501 TI - T-cell function is critical for murine cholesterol gallstone formation. AB - BACKGROUND & AIMS: The formation of cholesterol gallstones is a complex process involving contributions from genes and environmental factors. Although gallbladder inflammation is believed to be common during cholelithogenesis, the role of immunologic factors is unknown. METHODS: The role of adaptive immunity in cholesterol cholelithogenesis was analyzed utilizing immunocompetent Helicobacter spp.-infected and -uninfected BALB/c and congenic immunodeficient Rag2(-/-) (Rag) mice. Lymphocyte transfer studies were performed to determine which cellular subset was responsible for cholesterol gallstone formation. Also, gallbladder inflammation was quantified to determine the nature of the inflammatory response associated with cholelilithogenesis. RESULTS: When fed a lithogenic diet for 8 weeks, wild-type mice developed significantly more cholesterol gallstones (27% 80% prevalence) than Rag mice ( approximately 5%, P < .05). Helicobacter spp. infected BALB/cJ mice displayed statistically significant increases in cholesterol gallstone prevalence compared with uninfected mice (81% vs. 39%; P < .05). Transfer of splenocytes or T lymphocytes to Rag2(-/-) mice increased stone prevalence markedly (26% and 40% respectively; P < .05), whereas transfer of B cells was not appreciably cholelithogenic (13%). The adaptive immune response increased the expression of gallbladder Muc genes and accumulation of mucin gel. In addition, T cells and cholesterol monohydrate crystals induced proinflammatory gene expression in the gallbladder, which likely contributes to gallbladder dysfunction. CONCLUSIONS: These studies indicate that T cells are critical in murine cholesterol cholelithogenesis. Furthermore, cholesterol monohydrate crystals induce expression of proinflammatory cytokines in a T-cell-dependent fashion. Acquired immunity and inflammation are likely to be crucial factors in cholesterol gallstone pathogenesis, rather then merely the result of cholelithogenesis. PMID- 17919502 TI - Distinct Wilson's disease mutations in ATP7B are associated with enhanced binding to COMMD1 and reduced stability of ATP7B. AB - BACKGROUND & AIMS: Wilson's disease (WD) is characterized by hepatic copper overload and caused by mutations in the gene encoding the copper-transporting P type adenosine triphosphatase (ATPase) ATP7B. ATP7B interacts with COMMD1, a protein that is deleted in Bedlington terriers with hereditary copper toxicosis. Here we characterized the implications of the interaction between COMMD1 and ATP7B in relation to the pathogenesis of WD. METHODS: Glutathione-S-transferase pull-down experiments, co-immunoprecipitations, immunofluorescence microscopy, site-directed mutagenesis, and biosynthetic labeling experiments were performed to characterize the interaction between COMMD1 and ATP7B and the effects of WD causing mutations. RESULTS: COMMD1 specifically interacted with the amino terminal region of ATP7B. This interaction was independent of intracellular copper levels and of the expression of the copper chaperone ATOX1. Four WD patient-derived mutations in this region of ATP7B significantly increased its binding to COMMD1. Two of these mutations also resulted in mislocalization and increased degradation rate of ATP7B. Although COMMD1 did not affect copper induced trafficking of ATP7B, it markedly decreased the stability of newly synthesized ATP7B. CONCLUSIONS: Our data implicate COMMD1 in the pathogenesis of WD and indicate that COMMD1 exerts its regulatory role in copper homeostasis through the regulation of ATP7B stability. PMID- 17919503 TI - The genetics of inflammatory bowel disease. PMID- 17919504 TI - Eosinophilic esophagitis in children and adults: a systematic review and consensus recommendations for diagnosis and treatment. AB - During the last decade, clinical practice saw a rapid increase of patients with esophageal eosinophilia who were thought to have gastroesophageal reflux disease (GERD) but who did not respond to medical and/or surgical GERD management. Subsequent studies demonstrated that these patients had a "new" disease termed eosinophilic esophagitis (EE). As recognition of EE grew, so did confusion surrounding diagnostic criteria and treatment. To address these issues, a multidisciplinary task force of 31 physicians assembled with the goal of determining diagnostic criteria and making recommendations for evaluation and treatment of children and adults with suspected EE. Consensus recommendations were based upon a systematic review of the literature and expert opinion. EE is a clinicopathological disease characterized by (1) Symptoms including but not restricted to food impaction and dysphagia in adults, and feeding intolerance and GERD symptoms in children; (2) > or = 15 eosinophils/HPF; (3) Exclusion of other disorders associated with similar clinical, histological, or endoscopic features, especially GERD. (Use of high dose proton pump inhibitor treatment or normal pH monitoring). Appropriate treatments include dietary approaches based upon eliminating exposure to food allergens, or topical corticosteroids. Since EE is a relatively new disease, the intent of this report is to provide current recommendations for care of affected patients and defining gaps in knowledge for future research studies. PMID- 17919505 TI - Toward risk stratification for screening and surveillance of colorectal neoplasia: one small step for the colonoscopist. PMID- 17919506 TI - Would free Fatty acids enhance treatment of obesity? PMID- 17919507 TI - The cholinergic anti-inflammatory pathway and the gastrointestinal tract. PMID- 17919509 TI - Noninvasive markers of advanced histology in nonalcoholic fatty liver disease: are we there yet? PMID- 17919508 TI - It's about time: clock genes unveiled in the gut. PMID- 17919511 TI - Laryngopharyngeal reflux: beauty is in the eye of the beholder. PMID- 17919512 TI - Initial capsule endoscopy or angiography in patients with obscure gastrointestinal bleeding? PMID- 17919513 TI - Will virtual colonoscopy replace optical colonoscopy for colorectal cancer screening? PMID- 17919515 TI - Factors associated with incomplete colonoscopy: further information from a large prospective Italian survey. PMID- 17919516 TI - Are colonoscopies performed in private offices less likely to be successful? PMID- 17919517 TI - Assessing the extent of colonoscopy. PMID- 17919519 TI - Apoptosis-associated changes in the glycerophospholipid composition of hematopoietic progenitor cells monitored by 31P NMR spectroscopy and MALDI-TOF mass spectrometry. AB - Apoptosis, or programmed cell death, plays an important role in development and in tissue homeostasis and is assumed to be accompanied by changes in the composition of cellular glycerophospholipids (GPL). We have applied a combination of 31P nuclear magnetic resonance spectroscopy and matrix-assisted laser desorption and ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) to the analysis of organic extracts of hematopoietic progenitor cells undergoing the physiologically relevant process of apoptosis following growth factor withdrawal. The combined application of these methods enables the quantitative analysis of all glycerophospholipid classes and reveals changes in the acyl chain compositions from crude cell extracts. Using these techniques, an increase in the ratio of ether-linked glycerophospholipids to diacyl-glycerophospholipids during apoptosis was observed. The relative decrease in the membrane diacyl phosphatidylcholine (PC) levels was found to correlate with increased concentrations of the corresponding lysophosphatidylcholine (LPC) present in the medium. PMID- 17919520 TI - Letter from the editor: the fear of breast cancer. PMID- 17919522 TI - Case of the season: primary angiosarcoma of the breast: correlative imaging and pathology. PMID- 17919523 TI - Auditing your breast imaging practice: an evidence-based approach. PMID- 17919524 TI - Percutaneous breast biopsy: emerging techniques and continuing controversies. PMID- 17919525 TI - Breast magnetic resonance imaging: principles and techniques. PMID- 17919526 TI - Digital mammography: clinical implementation and clinical trials. PMID- 17919527 TI - Advanced applications of digital mammography: tomosynthesis and contrast-enhanced digital mammography. PMID- 17919528 TI - Lymph node status in the breast cancer patient: sampling techniques and prognostic significance. PMID- 17919529 TI - Positron emission tomography, positron emission tomography-computed tomography, and molecular imaging of the breast cancer patient. PMID- 17919530 TI - Computer-assisted detection in clinical practice: medical legal considerations. PMID- 17919531 TI - Performance characteristics and clinical utility of an enzymatic method for the measurement of glycated albumin in plasma. AB - OBJECTIVE: The measurement of plasma glycated albumin is particularly useful in the short-middle term monitoring of glycometabolic control in diabetics. The aim of this work is to evaluate a new enzymatic method for the measurement of glycated albumin in plasma, with particular attention to some selected cases and comparison with other relevant tests (fasting plasma glucose, after glucose load, fructosamine, glycated hemoglobin). DESIGN AND METHODS: We have performed a multicenter study by which sample collection was performed in three different centers (Milano, Padova and Cagliari) and serum samples, frozen at -80 degrees C, were then delivered under dry ice to the centralized laboratory in Milano. Glycated plasma albumin was measured with reagents from Asahi Kasei Pharma (Lucica GA-L enzymatic assay; AKP, Tokyo, Japan) on a Modular P Roche system. Fructosamine was assessed by a Roche method and HbA(1c) (measured separately in the three centers on fresh EDTA blood) by DCCT-aligned HPLC systems. We have investigated 50 type 2 diabetics, 26 subjects with gestational diabetes, 35 subjects with thalassemia major, 10 subjects with cirrhosis, 23 patients with end stage renal disease subjected to dialysis treatment and 32 healthy adult control subjects. RESULTS: The main analytical performance characteristics of the new GA test were the following: (a) the within-assay reproducibility was between 3.0 and 3.9% (in terms of GA% CV, measured on 2 serum pools and 2 control materials at normal and pathological glycated albumin levels); (b) the between-assays reproducibility was from 2.8 to 4.1%; (c) the linearity was tested in the interval between 13 and 36% and found acceptable (r(2)=0.9932). Concerning the clinical utility of the new test, we have evaluated the relationships between GA, HbA(1c), fructosamine and fasting and post-prandial glucose in several patients, as well as the changes in the above mentioned parameters in a sub-group of type 2 diabetic patients for 18 weeks as they progressed from severe hyperglycemia (HbA(1c) >or=10.0%) toward a better glycemic control. The correlations between glycated albumin and HbA(1c) were as follows: (a) type 2 diabetics: r(2)=0.483 (good glycemic control), r(2)=0.577 (poor control); (b) diabetic patients under dialysis: r(2)=0.480; (c) liver disease: r(2)=0.186; (d) transfused non-diabetics with thalassemia: r(2)=0.004. Glycated albumin, as well as HbA(1c) and fructosamine, was of little value in the study of women with gestational diabetes, mainly because of the very limited glucose fluctuations in this particular category of subjects. In 11 type 2 diabetic patients under poor metabolic control, GA was better correlated with fasting plasma glucose then HbA(1c) (r(2)=0.555 vs. 0.291, respectively), and decreased more rapidly than HbA(1c) during intensive insulin therapy. CONCLUSIONS: The experience we have acquired with the new enzymatic test demonstrates its reproducibility and robustness. We confirm that plasma glycated albumin is better related to fasting plasma glucose with respect to HbA(1c). Moreover, glycated albumin is more sensitive than HbA(1c) with regard to short-term variations of glycemic control during treatment of diabetic patients. This test is also very appropriate when the interpretation of HbA(1c) is critical. PMID- 17919532 TI - Thermal performance of quartz capillaries for vitrification. AB - In this paper we report the thermal behavior of a new approach for vitrification. Thermal performance of traditional open pulled straws is compared with a new technique based on the combined use of quartz capillaries with slush nitrogen. This new method of vitrification achieved ultrafast cooling rates of 250,000 degrees C/min. As a result, a much lower concentration of cryoprotectant was needed to reach vitrification. In fact, a cryoprotectant solution typically used in oocyte slow freezing protocols was shown to remain transparent after cooling to liquid nitrogen temperatures indicating apparent "vitrification". This approach offers a new and very promising technique for vitrification of cells using low levels of cryoprotectants. PMID- 17919533 TI - TCF4 deficiency expands ventral diencephalon signaling and increases induction of pituitary progenitors. AB - The anterior and intermediate lobes of the pituitary gland are formed from Rathke's pouch. FGF, BMP and WNT signals emanating from the ventral diencephalon influence pouch growth and development. In order to examine the role of canonical WNT signaling during pituitary development we examined the pituitary expression of the TCF/LEF family of transcription factors, which mediate WNT signaling through the binding of beta-catenin. We report here the expression of several members of this family during pituitary development and the functional role of one member, TCF4 (TCF7L2), in the induction of the pituitary primordium. TCF4 is expressed in the ventral diencephalon early in pituitary development, rostral to a domain of BMP and FGF expression. Tcf4 deficient mice express Fgf10 and Bmp4; however, the Bmp and Fgf expression domains are expanded rostrally. As a result, additional pituitary progenitor cells are recruited into Rathke's pouch in Tcf4 mutants. Mutants also exhibit an expansion of the Six6 expression domain within Rathke's pouch, which may increase the number of proliferating pouch cells, resulting in a greatly enlarged anterior pituitary gland. This suggests that TCF4 negatively regulates pituitary growth through two mechanisms. The first mechanism is to restrict the domains of BMP and FGF signaling in the ventral diencephalon, and the second mechanism is the restriction of Six6 within Rathke's pouch. Thus, TCF4 is necessary both intrinsically and extrinsically to Rathke's pouch to ensure the proper growth of the pituitary gland. PMID- 17919534 TI - Laminin alpha5 is essential for the formation of the zebrafish fins. AB - The vertebrate fin fold, the presumptive evolutionary antecedent of the paired fins, consists of two layers of epidermal cells extending dorsally and ventrally over the trunk and tail of the embryo, facilitating swimming during the embryonic and larval stages. Development of the fin fold requires dramatic changes in cell shape and adhesion during early development, but the proteins involved in this process are completely unknown. In a screen of mutants defective in fin fold morphogenesis, we identified a mutant with a severe fin fold defect, which also displays malformed pectoral fins. We find that the cause of the defect is a non sense mutation in the zebrafish lama5 gene that truncates laminin alpha5 before the C-terminal laminin LG domains, thereby preventing laminin alpha5 from interacting with its cell surface receptors. Laminin is mislocalized in this mutant, as are the membrane-associated proteins, actin and beta-catenin, that normally form foci within the fin fold. Ultrastructural analysis revealed severe morphological abnormalities and defects in cell-cell adhesion within the epidermis of the developing fin fold at 36 hpf, resulting in an epidermal sheet that can not extend away from the body. Examining the pectoral fins, we find that the lama5 mutant is the first zebrafish mutant identified in which the pectoral fins fail to make the transition from an apical epidermal ridge to an apical fold, a transformation that is essential for pectoral fin morphogenesis. We propose that laminin alpha5, which is concentrated at the distal ends of the fins, organizes the distal cells of the fin fold and pectoral fins in order to promote the morphogenesis of the epidermis. The lama5 mutant provides novel insight into the role of laminins in the zebrafish epidermis, and the molecular mechanisms driving fin formation in vertebrates. PMID- 17919535 TI - The Drosophila Dead end Arf-like3 GTPase controls vesicle trafficking during tracheal fusion cell morphogenesis. AB - The Drosophila larval tracheal system consists of a highly branched tubular organ that becomes interconnected by migration-fusion events during embryonic development. Fusion cells at the tip of each branch guide migration, adhere, and then undergo extensive remodeling as the tracheal lumen extends between the two branches. The Drosophila dead end gene is expressed in fusion cells, and encodes an Arf-like3 GTPase. Analyses of dead end RNAi and mutant embryos reveal that the lumen fails to connect between the two branches. Expression of a constitutively active form of Dead end in S2 cells reveals that it influences the state of actin polymerization, and is present on particles that traffic along actin/microtubule containing processes. Imaging experiments in vivo reveal that Dead end-containing vesicles are associated with recycling endosomes and the exocyst, and control exocyst localization in fusion cells. These results indicate that the Dead end GTPase plays an important role in trafficking membrane components involved in tracheal fusion cell morphogenesis and lumenal development. PMID- 17919536 TI - Myogenic reprogramming of retina-derived cells following their spontaneous fusion with myotubes. AB - Satellite cells are recognized as the main source for myoblasts in postnatal muscle. The possible participation of other cell types in myofiber maintenance remains a subject of debate. Here, we investigated the potential of vascular preparations from mouse retina to undergo myogenesis when cultured alone or with differentiated primary myogenic cultures. The choice of retina, an organ richly supplied with capillary network and anatomically separated from skeletal muscles, ensures that the vasculature preparation is devoid of satellite cells. We demonstrate that retina-derived cells spontaneously fuse with preexisting myotubes and contribute additional myonuclei, some of which initiate expression of muscle-specific genes after fusion. Myogenic differentiation of retinal cells prior to their fusion with preexisting myotubes was not detected. Although originating from vasculature preparations, nuclei undergoing myogenic reprogramming were contributed by cells that were neither endothelial nor blood borne. Our results suggest smooth muscle/pericytes as the possible source, and that myogenic reprogramming depends on the muscle specific transcription factor MyoD. Our studies provide insights into a novel avenue for myofiber maintenance, relying on nuclei of non-myogenic origin that undergo fusion and subsequent myogenic conversion within host myofibers. This process may support ongoing myofiber maintenance throughout life. PMID- 17919537 TI - Reducing the risk for breast cancer recurrence after completion of tamoxifen treatment in postmenopausal women. AB - BACKGROUND: In postmenopausal women with hormone-sensitive early stage breast cancer, the risk for relapse persists after 5 years of treatment with adjuvant tamoxifen. Because tamoxifen is not indicated for adjuvant therapy beyond 5 years, the need for another therapy in the extended adjuvant setting to reduce late recurrence risk is clear. The National Cancer Institute of Canada Clinical Trials Group MA.17 and the National Surgical Adjuvant Breast and Bowel Project (NSABP) B-33 trial found that extended adjuvant therapy with an aromatase inhibitor (AI) (eg, letrozole, exemestane, or anastrozole) rendered additional benefit in postmenopausal women with hormone receptor-positive breast cancer. The MA.17 trial was unblinded at the first interim analysis (median follow-up, 2.4 years) due to a significant reduction in relative risk for recurrence (P < 0.001). Following the publication of the final analysis, several other MA.17 trial analyses and a postblinding analysis were also conducted. Recent data on the NSABP B-33 trial, which investigated exemestane in the extended adjuvant setting, and Austrian Breast and Colorectal Cancer Study Group (ABCSG) trial 6a, which evaluated anastrozole in the extended adjuvant setting, have also been reported. OBJECTIVE: The goal of this article was to provide a current review of the MA.17 and NSABP B-33 results, together with additional data, to determine the benefit and tolerability of extended adjuvant AI therapy and the potential benefits of late extended adjuvant therapy with AIs after a prolonged (or=60 years. Among the 23 patients whose sex was reported, only 3 (13%) were men. Of 15 patients with a history of invasive dental treatment, 12 (80%) had undergone dental surgery or experienced dental trauma at the site of ONJ. Among the 10 patients for whom the duration of bisphosphonate treatment was reported, no clear relationship between the duration of bisphosphonate treatment and the development of ONJ was observed. CONCLUSIONS: Considering that millions of patients have been prescribed bisphosphonates for the treatment of osteoporosis, the relative prevalence of ONJ in these patients was low. Age >or=60 years, female sex, and previous invasive dental treatment were the most common characteristics of those who developed ONJ. However, it is not possible to draw further conclusions about the potential association between oral bisphosphonate use and ONJ in the identified studies because of incomplete reporting and the presence of confounding factors. PMID- 17919539 TI - Darunavir: a nonpeptidic antiretroviral protease inhibitor. AB - BACKGROUND: Protease inhibitors were a major therapeutic breakthrough in the mid 1990s for the treatment of HIV infection, which resulted in improved life expectancy for patients who had failed previous therapies. With time and evolution of the virus, however, there is a new population of patients with treatment-resistant disease and few treatment options. Darunavir is a synthetic nonpeptidic analogue of amprenavir with enhanced activity against resistant virus that became available in 2006. OBJECTIVES: The purpose of this review was to describe the clinical pharmacology, pharmacokinetic and pharmacodynamic properties, and clinical efficacy of darunavir. Also discussed are the published clinical experience with darunavir, its adverse events, drug interactions, pharmacoeconomics, and dosing and administration. METHODS: A MEDLINE and EMBASE search (English-language only) was performed from January 1996 through April 2007 using the key words darunavir and TMC114. Abstracts from relevant scientific meetings were searched for the years 2000 through 2007. Additionally, the US Food and Drug Administration Web site was accessed to review the new drug application summary and data presented therein. RESULTS: Darunavir was found to maintain antiretroviral activity against HIV with protease inhibitor mutations in 6 studies. Clinical efficacy and safety data are limited to 4 controlled and 2 uncontrolled trials. In 2 large Phase IIb clinical studies, viral suppression at 48 weeks to undetectable levels in heavily pretreated patients was achieved in 45% of patients compared with 10% of patients in the control group (P < 0.001). The addition of enfuvirtide enhanced this response rate to 58% compared with 11% of the patients who did not receive enfuvirtide (P < 0.001). Gastrointestinal symptoms, nausea, and headache were the most commonly reported events. CONCLUSIONS: Darunavir has improved activity against resistant HIV isolates in patients with few treatment choices, particularly when enfuvirtide is added. The safety profile of darunavir is comparable to other protease inhibitors based on early data. PMID- 17919541 TI - Efficacy and safety of inhaled zanamivir in the prevention of influenza in community-dwelling, high-risk adult and adolescent subjects: a 28-day, multicenter, randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: Influenza can cause significant morbidity and mortality in subjects at high risk for complications, including the elderly (age >or=65 years) and those with chronic respiratory, cardiovascular, or metabolic conditions. Effective prophylaxis can significantly reduce the disease burden in this population. Previous studies conducted primarily in non-high-risk subjects have reported the efficacy of inhaled zanamivir in preventing influenza. OBJECTIVE: This study investigated the efficacy and safety of zanamivir in preventing influenza in community-dwelling adult and adolescent subjects at high risk for complications of influenza. METHODS: This was a multicenter, randomized, double blind, placebo-controlled, parallel-group study in community-dwelling subjects aged >or=12 years who were at high risk for developing complications of influenza, were able to use the Diskhaler device (Glaxo Group Limited, Research Triangle Park, North Carolina), and were able to take the first dose of study medication within 5 days of laboratory-confirmed local influenza activity. Eligible subjects were randomized to receive inhaled zanamivir 10 mg or placebo once daily for 28 days. The primary end point was the proportion of randomized subjects who developed symptomatic influenza during prophylaxis, as confirmed by culture and/or serology. All adverse events (AEs) occurring after the first dose of study medication were recorded. RESULTS: The study enrolled 3363 subjects, of whom 58% were female and 93% were white; the mean age of participants was 60.4 years (range, 12-94 years), and 4% were adolescents. Significantly fewer zanamivir-treated subjects developed symptomatic, laboratory- confirmed influenza during prophylaxis compared with placebo recipients (4/1678 vs 23/1685, respectively), representing a relative risk (RR) of 0.17 (95% CI, 0.07-0.44; P < 0.001) and a protective efficacy of 83%. The incidence of complications was reduced in zanamivir-treated subjects compared with placebo recipients (1/1678 and 8/1685), representing an RR of 0.12 (95% CI, 0.02-0.73; P = 0.042) and a protective efficacy of 88%. The numbers of zanamivir recipients (151/1678 [9%]) and placebo recipients (169/1685 [ 10 % ] ) who developed symptomatic influenza like illness regardless of laboratory confirmation did not differ significantly (RR = 0.86; 95% CI, 0.70-1.06), indicating that zanamivir was not effective in preventing influenza-like illness that was not caused by influenza infection. Similarly, there was no significant difference in the numbers of zanamivir and placebo recipients who developed laboratory-confirmed infection regardless of symptoms (39/1678 [2%] and 52/1685 [3%], respectively; RR = 0.76; 95% CI, 0.50 1.15). Of these, 64 subjects (35 and 29) were asymptomatic; seroconversion occurred in all but 1 subject, indicating that zanamivir prophylaxis did not prevent asymptomatic seroconversion. During prophylaxis, 51% of subjects in both treatment groups reported at least 1 AE. There were no major differences in the frequency or nature of AEs between groups. The most commonly reported AEs (>or=3% of subjects in each treatment group) were consistent with upper respiratory viral infection (headache: 17% zanamivir, 18% placebo; cough: 14% and 15%, respectively; throat and tonsil discomfort/pain: 13% and 14%). There were no differences between groups in the overall incidence of viral respiratory infections (5% in both groups) or ear, nose, and throat infections (2% in both groups). None of the analyzed isolates from confirmed cases of influenza exhibited reduced susceptibility to zanamivir or genotypic evidence of resistance. CONCLUSIONS: Zanamivir, administered once daily for 28 days, was efficacious in preventing infection with the predominant circulating strains in the 2000- 2001 influenza season in the Northern Hemisphere (influenza A/New Calendonia/20/99-1ike and influenza B/ Sichuan/379/99-like) in these high-risk community- dwelling subjects aged >or=12 years. Zanamivir was well tolerated, with a safety profile comparable to that of placebo. No emergence of resistant virus was detected. PMID- 17919542 TI - Transdermal buprenorphine patches applied in a 4-day regimen versus a 3-day regimen: a single-site, Phase III, randomized, open-label, crossover comparison. AB - BACKGROUND: In 2001, a transdermal matrix patch formulation of buprenorphine was approved for the treatment of moderate to severe cancer pain and severe pain that is unresponsive to nonopioid analgesics. The primary recommendation contained in the prescribing information was that transdermal patches be worn for a 3-day period before application of a new patch. OBJECTIVE: This study was conducted to evaluate the potential for extending the time the buprenorphine patch is worn from 3 to 4 days. METHODS: This single-center, randomized, open-label, crossover Phase III study compared the efficacy and tolerability of the buprenorphine transdermal patch applied for different durations, with patch changes every 3 days versus every 4 days (12 days each), in patients with chronic moderate or severe pain of malignant or nonmalignant origin. Study participants were aged >18 years, had already responded to at least 4 weeks of transdermal buprenorphine, and had achieved steady-state conditions for at least 2 weeks before enrollment. The primary end point was patients' rating of the quality of treatment (analgesic efficacy and tolerability, rated on a 5-point scale: very good, good, satisfactory, poor, and inadequate) at the completion of each treatment regimen. Also recorded were physicians' ratings of the quality of treatment; pain intensity, rated on an 11-point numerical rating scale (from 0 = no pain to 10 = worst pain imaginable) and on the McGill Pain Questionnaire (MPQ) (maximum pain = 3.0); health status, assessed using the 36-item Short Form Health Survey (SF-36), expressed as a percentage of the best health condition (100%); and pain relief (5 point scale: complete, good, satisfactory, slight, and none). Local skin tolerability was evaluated for objective and subjective dermatologic symptoms at the patch application sites. Patients recorded daily pain intensities at specified times of day and night, pain relief (5-point verbal rating scale), and sleep duration (2-3 hours, >3-<6 hours, or >or=6 hours) in a diary. The safety profile was evaluated based on standard monitoring of adverse events, vital signs, and routine laboratory tests. RESULTS: Forty-nine white patients (25 women, 24 men) were enrolled; their mean (SD) age was 61.6 (11.5) years, and their mean weight was 74.7 (16.7) kg. The most common source of pain was musculoskeletal disorders (40 patients), followed by nervous system disorders (10), neoplasms (9), injuries (5), and other causes (6). Forty-one patients completed the study; 2 patients discontinued because of adverse events, 1 because of lack of efficacy, and 5 for nonmedical reasons. Thirty-three patients provided data per protocol. Patients in the perprotocol population received a mean (SD) transdermal buprenorphine dose of 49.9 (38.9) microg/h. The proportion of patients in the per-protocol population rating the quality of treatment as adequate (combined ratings of very good, good, and satisfactory) was 93.9% (31/33) for both regimens. The physicians' ratings indicated adequate quality of treatment in 93.8% (30/32) of patients applying 4 patches for 3 days each and 97.0% (32/33) of patients applying 3 patches for 4 days each. Mean (SD) pain intensity scores on the numerical rating scale were similar after completion of the 3- and 4-day regimens (3.73 [1.88] and 3.88 [1.75] points, respectively), as were MPQ scores (0.79 [0.67] and 0.79 [0.78]). The mean (SD) proportion of days with at least satisfactory pain relief was 83.9% (26.1%) and 85.6% (24.4%) for the 3- and 4-day regimens; the corresponding proportions of nights with at least satisfactory pain relief were 85.2% (26.6%) and 88.1% (21.4%). Continuously assessed pain intensities at specified times of day and night (numerical rating scale) did not differ significantly between regimens. Mean SF-36 health status scores did not differ significantly between regimens (total score: 37.7% [17.0%] and 37.7% [17.3%]). Mean rates of nights with good sleep quality were 28.5% (39.9%) for the 3-day regimen and 36.0% (42.6%) for the 4-day regimen. Local skin tolerability was comparable for the 3- and 4-day regimens, with objective findings (mainly erythema) at the patch-application sites in 17 of 32 and 11 of 33 patients, respectively, and subjective symptoms (mainly itching) in 16 of 32 and 13 of 33 patients. The most common adverse events in the safety population were nausea, dizziness/giddiness, and malaise/fatigue (3/49 [6.1%] each). CONCLUSION: Analgesic efficacy, patients' satisfaction with the quality of treatment, and skin tolerability did not differ significantly between 3 and 4 days of patch application in these patients with chronic pain who had been previously stabilized on transdermal buprenorphine. PMID- 17919543 TI - Negative binomial meta-regression analysis of combined glycosylated hemoglobin and hypoglycemia outcomes across eleven Phase III and IV studies of insulin glargine compared with neutral protamine Hagedorn insulin in type 1 and type 2 diabetes mellitus. AB - OBJECTIVES: This analysis first modeled the interaction between hypoglycemia and glycosylated hemoglobin (HbA1c) in clinical trials that compared insulin glargine (glargine) with human neutral protamine Hagedorn insulin (NPH) in patients with type 1 or type 2 diabetes mellitus. The model was then used to compare rates of hypoglycemia associated with use of these insulins. METHODS: Patient-level data from all randomized Phase III/IV clinical trials sponsored by the manufacturer of glargine that compared glargine and NPH and were available in May 2004 were included in the model. In addition, MEDLINE, EMBASE, and BIOSIS were searched for comparative randomized controlled trials of glargine and NPH using the terms insulin glargine, HOE 901, neutral protamine Hagedorn insulin, and NPH insulin. Studies were excluded from the analysis if patient-level data were not available. Unadjusted rates of symptomatic, confirmed, and severe hypoglycemia were compared with those derived from negative binomial regression analysis, which stratified the results by HbA1c at end point (with last observation carried forward), treatment, and duration of diabetes. In addition, the analysis was stratified by Phase III studies (which focused on determining tolerability and efficacy before regulatory approval) and Phase IV studies (which compared the clinical efficacy of the 2 insulins). The first month of the study was not included in the analysis because of continual adjustment of the insulin dose and maintenance of previous NPH in some studies. RESULTS: Eleven sponsored randomized trials were included in the model (total of 5074 patients). Four other sponsored trials were not included because the databases were not finalized, and 3 investigator-initiated trials were not included because patient-level data were unavailable. Rates of hypoglycemia had a curvilinear relationship with HbAlc, increasing at lower end point HbAlc values. In combined analyses of the studies of type 1 and type 2 diabetes, unadjusted rates of hypoglycemia were lower for glargine than NPH: 6.1% lower for all symptomatic hypoglycemia, 21.6% lower for confirmed hypoglycemia, and 23.9% lower for severe hypoglycemia (all, P < 0.05). When modeled using the negative binomial distribution with end-point HbA1c as a covariate, the corresponding results were 9.1% (P < 0.05), 26.6% (P < 0.001), and 30.0% (P = 0.08), respectively. When only Phase IV trials were analyzed, the relative reductions with glargine were 16.2% (P < 0.01), 40.8% (P < 0.01), and 46.8% (P < 0.05). The results of the separate analyses of studies of type 1 and type 2 diabetes were comparable. CONCLUSIONS: Based on the results of this analysis, calculated unadjusted hypoglycemia event rates appear to underestimate the differences between glargine and NPH. In most of the present analyses, unadjusted rates were significantly lower with glargine than NPH. Adjustment for end-point HbA1c resulted in greater relative reductions in the risk of hypoglycemia for glargine compared with NPH. The adjusted risk reduction with glargine was highest in the Phase IV studies. PMID- 17919544 TI - Pharmacokinetic and pharmacodynamic properties of buprenorphine after a single intravenous administration in healthy volunteers: a randomized, double-blind, placebo-controlled, crossover study. AB - BACKGROUND: Buprenorphine is used as an analgesic for postoperative and chronic pain. The usual sublingual dose is 0.2 to 0.8 mg, and the usual parenteral dose is 0.3 mg for acute postoperative pain. The pharmacokinetic and related pharmacodynamic properties of buprenorphine at these doses have not been characterized. OBJECTIVE: The aim of this study was to assess the pharmacokinetic properties of buprenorphine 0.002 mg/kg IV (0.15 mg/70 kg) and its antinociceptive and psychomotor effects. METHODS: Healthy male volunteers received 0.002 mg/kg buprenorphine IV in a randomized, double-blind, placebo controlled, crossover design. Blood samples were collected at 0.5, 1, 1.5, 1.75, 2, 2.5, 3, 4, 5, 6, and 8 hours for the determination of plasma concentrations. Pharmacokinetic parameters were estimated by a compartmental model using specialized software. Antinociceptive and psychomotor effects were determined for 8 hours. Quantitative sensory testing with thermal and electrical (nociceptive flexion RIII reflex) stimulations was performed. The cold pressor test was used to assess pain tolerance to a tonic, intense pain stimulation. Psychomotor performance was assessed by the digit symbol substitution test (DSST). Participants also rated sedation on an 11-point numeric scale (0 = none to 10 = severe). A selective liquid chromatography-tandem mass spectrometry assay was developed for the determination of buprenorphine; the limit of quantification was 0.05 ng/mL using a 0.25-mL plasma aliquot. Participants were instructed to report adverse effects, which were recorded for type, time of onset, seriousness, and duration. RESULTS: The study enrolled 12 participants, all of whom were white. Mean (SD) age was 26 (3.5) years, and mean weight was 67 (9) kg. None of the participants had a history of opiate abuse. Buprenorphine significantly increased the objective (nociceptive flexion RIII reflex) and subjective pain thresholds for >4 hours and pain tolerance (cold pressor test) for 2 hours. The mean (SD) RIII reflex threshold and subjective threshold at baseline were 31.6 (9.5) mA and 45.5 (22.3) mA, respectively. The maximum increases (mean [SD]) were +14.1 (17.5) mA for the RIII reflex (P = 0.02) and +24.2 (21.7) mA for the subjective threshold (P = 0.02), corresponding to mean (SEM) percentages of 53.7% (20.2%) and 74.7% (20.4%) of the baseline values, respectively. The maximum increases were observed at 120 minutes for both measures. The effect of buprenorphine on pain tolerance peaked at 30 minutes. Mean (SEM) latency before withdrawal of the hand was 69 (10) seconds, corresponding to a mean increase of 63.8% (14.4%) from baseline (P = 0.003). Buprenorphine had a significant effect on the DSST. The mean maximum decrease in the total number of symbols drawn was -6 (14.5%; P = 0.005) at 1 hour. The participants reported high levels of sedation: at peak effect (120 minutes), mean scores increased from 2.9 to 6.4 (SEM 0.7) (P = 0.005). Levels returned to baseline values by the end of the session, unlike for the nociceptive tests. The onset of effects occurred during the distribution phase for all the measures, and their duration was observed across a wide range of concentrations during the elimination phase. The most likely explanation for this finding is the high affinity of buprenorphine at mu-opioid receptors, and possibly distribution to the brain. Buprenorphine t(l/2) was 2.75 hours. A secondary peak in concentration was observed at 90 minutes, suggesting enterohepatic circulation of buprenorphine. A 2-compartment model adequately described buprenorphine pharmacokinetics. CONCLUSIONS: A clinically relevant analgesic dose of 0.002 mg/kg (0.15 mg/70 kg) of buprenorphine had a significant effect on nociception and psychomotor performance in these healthy male volunteers. A 2-compartment model satisfactorily characterized buprenorphine pharmacokinetics, and we found evidence of enterohepatic circulation. PMID- 17919545 TI - Oral garenoxacin in the treatment of acute bacterial maxillary sinusitis: a Phase II, multicenter, noncomparative, open-label study in adult patients undergoing sinus aspiration. AB - BACKGROUND: Garenoxacin is a des-F(6)-quinolone with in vitro activity against key respiratory pathogens, including Streptococcus pneumoniae, Hemophilus influenzae, Staphylococcus aureus, and Moraxella catarrhalis. Limited data are available regarding the effect of garenoxacin in the treatment of acute bacterial sinusitis. OBJECTIVE: The aim of this study was to assess the efficacy and tolerability of garenoxacin in adults with acute bacterial maxillary sinusitis undergoing a pre-treatment diagnostic sinus aspirate. METHODS: This Phase II, multicenter, noncomparative, open-label study was conducted at 30 centers in the United States, Mexico, Argentina, and Europe. Male and female patients aged 18 to 80 years with clinical signs and symptoms lasting >or=5 but or=5 mm because it was believed that improvement in mucosal thickening might not be reliably measurable at the 5 day time point. All patients received garenoxacin 400 mg QD for 5 or 10 days. Maxillary sinus needle aspiration for Gram stain, routine culture, and susceptibility testing were performed before treatment, and, if clinically indicated, during and after treatment. Bacteriologic eradication (negative culture on repeat sinus aspiration) and cure rates (complete resolution of all signs and symptoms) were assessed at a test-of-cure visit 5 to 18 days after the end of treatment. The occurrence of adverse events was recorded by the investigators up to 30 days after the last administration of garenoxacin by questioning patients. RESULTS: A total of 546 patients were enrolled and 543 were randomized (5-day cohort: mean age, 40 years; mean weight, 76 kg; 56% women; 10 day cohort: mean age, 41 years; mean weight, 77 kg; 58% women). Clinically evaluable patients included 253 in the 5-day cohort and 266 in the 10-day cohort. Cure rates were 93% (236/253; 95% CI, 89%-96%) and 91% (243/266; 95% CI, 87%-94%) for evaluable patients in the 5- and 10-day cohorts, respectively. Bacteriologic eradication rates in microbiologically evaluable patients were 94% in both cohorts (5 days, 204/217; 10 days, 182/193). Eradication rates in the 5- and 10 day cohorts were as follows: S pneumoniae, 94% (62/66) and 93% (39/42); H influenzae, 100% (30/30) and 93% (26/28); S aureus, 96% (23/24) and 91% (31/34); and M catarrhalis, 89% (8/9) and 86% (12/14). Of the 9 patients with acute bacterial sinusitis due to multidrug-resistant S pneumoniae, 8 achieved clinical cure with garenoxacin treatment. Adverse events (AEs) most frequently reported were diarrhea (or=1 prescription for these medications in the 6-month periods before and after the date of the first gabapentin or pregabalin prescription), and attainment of therapeutic dose levels (gabapentin, >or=1800 mg/d; pregabalin, >or=150 mg/d). RESULTS: The database search identified 151 patients with PHN who were newly prescribed gabapentin (57.0% female; mean [SD] age, 55.8 [11.3] years) and 100 patients who were newly prescribed pregabalin (62.0% female; mean age, 52.8 [9.4] years). The prevalence of comorbidities did not differ significantly between recipients of prescriptions for gabapentin or pregabalin, with the exception of hyperlipidemia, which was more prevalent in those prescribed gabapentin (33.8% vs 22.0%, respectively; P = 0.044), and depression, which was more prevalent in those prescribed pregabalin (12.0% vs 4.6%, respectively; P = 0.031). In the pretreatment period, those who were prescribed pregabalin had significantly greater use of long-acting opioids (P = 0.005), anticonvulsants (P < 0.001), selective norepinephrine reuptake inhibitors (P < 0.001), and the lidocaine 5% patch (P = 0.005) compared with those prescribed gabapentin. Use of any opioid increased from pretreatment to follow-up in those prescribed gabapentin, significantly so in those who received >or=2 opioid prescriptions (P = 0.016). Use of any opioid decreased significantly from pre-treatment to follow up in those prescribed pregabalin (P = 0.005), particularly in those who received >or=2 opioid prescriptions (P = 0.004). Tramadol use decreased significantly in those prescribed gabapentin (P = 0.045), and anticonvulsant use decreased significantly in those prescribed pregabalin (P = 0.004). Among patients prescribed gabapentin or pregabalin who received 1 prescription, 2 consecutive prescriptions, and >or=3 consecutive prescriptions, a greater proportion of those prescribed pregabalin attained therapeutic dose levels by their first, second, and third consecutive prescriptions compared with those prescribed gabapentin (69.0% vs 3.5%, respectively [P < 0.001]; 71.4% vs 21.7% [P < 0.001]; and 89.3% vs 46.2% [P < 0.001], respectively). CONCLUSIONS: In these patients with PHN in the usual-care setting, opioid use increased after the initiation of gabapentin and decreased after the initiation of pregabalin. Few of those prescribed gabapentin received a prescription for a therapeutic dose, whereas a greater proportion of patients who were prescribed pregabalin received a prescription for a therapeutic dose. PMID- 17919548 TI - Mixed dyslipidemia among patients using lipid-lowering therapy in French general practice: an observational study. AB - BACKGROUND: Low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglycerides (TGs) are associated with an increased incidence of coronary heart disease (CHD). However, limited data are available about the prevalence of dyslipidemias related to LDL-C, HDL-C, and TGs among French patients treated with lipid-lowering agents. OBJECTIVE: This paper describes the prevalence of various types of dyslipidemias among patients treated with lipid-lowering agents in French general practice. METHODS: This was a cross sectional, observational study conducted using retrospective data collection at the time of enrollment. Eligible patients were those treated pharmacologically for dyslipidemia in the Cegedim Strategic Data general practice network. Fasting lipid values and cardiovascular (CV) risk factors were gathered by investigators using an ad hoc questionnaire. European guidelines were used to define various types of dyslipidemias. Polytomous logistic regression was used to assess the associations between different dyslipidemias and diabetes mellitus, a history of CHD, and the number of CV risk factors. RESULTS: A total of 946 patients had a complete lipid profile and valid data for determining CV risk status. The mean (SD) age of these patients was 64.0 (9.9) years, and 55.7% of the patients were men. At least 1 abnormality in LDL-C, HDL-C, or TGs was present in 791 (83.6 %) of the 946 patients. The rates of elevated LDL-C, low HDL-C, and elevated TGs were 73.2%, 16.9%, and 30.3%, respectively (these groups are not mutually exclusive). Among those who did not reach the LDL-C goal, 38.7% had dyslipidemias with low HDL-C, elevated TGs, or both. Compared with having a normal lipid profile, each additional CV risk factor increased the likelihood of the following types of dyslipidemias: low HDL-C and/or elevated TGs, but normal LDL-C (odds ratio [OR], 1.36; 95% CI, 1.03-1.79); elevated LDL-C and TGs, but normal HDL-C (OR, 1.58; 95% CI, 1.24-2.02); and all 3 lipid abnormalities (OR, 1.54; 95% CI, 1.10-2.14). Patients with diabetes had a similarly increased risk of mixed dyslipidemias, whereas patients with a history of CHD did not. CONCLUSIONS: Among these patients treated with lipid-lowering agents, 38.7% had mixed dyslipidemias, including low HDL-C, elevated TGs, both low HDL-C and elevated TGs, or all 3 lipid abnormalities. Patients with a greater number of nonlipid CV risk factors or with diabetes had a significantly increased risk of mixed dyslipidemias involving elevated TGs and/or low HDL-C in addition to elevated LDL-C. PMID- 17919549 TI - Goserelin acetate 10.8 mg plus iron versus iron monotherapy prior to surgery in premenopausal women with iron-deficiency anemia due to uterine leiomyomas: results from a Phase III, randomized, multicenter, double-blind, controlled trial. AB - BACKGROUND: Women with symptomatic uterine leiomyomas (fibroids) may have iron deficiency anemia (IDA); therefore, surgery places them at risk of blood-borne morbidity from perioperative transfusions. Such women might benefit from a preoperative treatment that restores hematologic normality and alleviates fibroid symptoms. OBJECTIVE: The purpose of this study was to examine the effects of a single preoperative depot injection of goserelin acetate plus iron treatment compared with iron monotherapy, in premenopausal women with IDA due to uterine leiomyomas. METHODS: This Phase III, randomized, multicenter, double-blind, controlled trial (12 weeks of treatment plus a 24-week follow-up period) was conducted from October 1997 to August 1999. Patients received an injection of goserelin acetate 10.8 mg (3-month formulation) or a sham, with both groups receiving PO iron (ferrous sulfate) 325-mg tablets TID during the 12-week treatment period. Surgery (hysterectomy or myomectomy) was planned for week 12. Hemoglobin (Hb) level, symptoms of uterine leiomyomas, requirement for blood transfusion throughout, ability to donate blood for autologous transfusion, and leiomyoma and uterine volume were assessed for efficacy. The tolerability assessment included bone mineral density measurements and subjective symptomatology (ie, menstrual bleeding [uterine hemorrhage], fatigue, pelvic pain, and pelvic pressure). RESULTS: A total of 110 women received treatment (n = 54, goserelin acetate 10.8 mg; n = 56, sham). The majority of patients (69.1%) were black and the mean age at study entry was 39.9 years, with a mean weight of 80.1 kg. At approximately 12 weeks, Hb levels were significantly higher in the goserelin group compared with the sham group (difference of least squares mean, 1.17 g/dL; 95% CI, 0.68-1.66; P < 0.001), and significantly more patients in the goserelin group had an increase in Hb concentration of >or=2 g/dL (odds ratio 6.36; 95% CI, 2.00-20.18; P < 0.001). A nonsignificant decrease in both uterine and leiomyoma volume was experienced by patients who administered goserelin compared with increases in the sham group. Uterine hemorrhage was also experienced numerically less often by goserelin-treated patients compared with those given the sham injection (9.3% vs 28.6%, respectively). One or more adverse events (AEs) were reported by 89% of patients in each treatment group. Goserelin acetate 10.8 mg was generally well tolerated by patients, with no serious drug related AEs reported during this 36-week trial. CONCLUSION: A single, preoperative injection of goserelin acetate 10.8 mg in addition to PO iron 325 mg TID was associated with improved Hb levels in these premenopausal women with IDA due to uterine leiomyomas. PMID- 17919550 TI - Pharmacokinetic and pharmacodynamic assessments of the dipeptidyl peptidase-4 inhibitor PHX1149: double-blind, placebo-controlled, single- and multiple-dose studies in healthy subjects. AB - BACKGROUND: PHX1149 is a dipeptidyl peptidase-4 (DPP4) inhibitor that is currently in clinical development for the treatment of type 2 diabetes mellitus. PHX1149 is a small (molecular weight = 241.16 Da), highly water-soluble (>2 g/mL), orally active molecule with a selectivity index of 15- to 319-fold relative to those of other members of the DPP family. The biochemical median inhibitory concentration of DPP4 is 2.5 nmol/L. OBJECTIVE: The aim of these 2 double-blind, randomized, placebo-controlled studies was to examine the pharmacokinetic (PK) parameters and pharmacodynamic (PD) properties and tolerability of single and multiple ascending doses of PHX1149 in healthy human subjects. METHODS: Healthy men and women aged 18 to 60 years were recruited to participate in a single- or a multiple-dose study in which sequential dose escalation paradigm was used. In the single-dose study, subjects were given a single oral dose of PHX1149 50 to 500 mg or placebo; in the multiple-dose study, subjects were given PHX1149 at doses from 50 to 400 mg or placebo QD for 13 days. There was no intrasubject dose escalation. Blood samples were collected from each subject at a series of time points ranging from 1 hour before to 24 hours after dosing on day 1 in the single- dose study and on days 1, 7, and 13 in the multiple-dose study. PK and PD analyses were performed in plasma samples to determine Cmax, Tmax, AUC0-t, AUC0-infinity, and DPP4 enzymatic activity. The drug accumulation index was also calculated for each dose of PHX1149 in the multiple-dose study. Adverse events (AEs) were monitored in both studies through physical examinations, including measurement of vital signs, and clinical laboratory testing. In both studies, electrocardiography was performed. RESULTS: The single- and multiple-dose studies enrolled 30 and 28 subjects, respectively, for a total enrollment in the 2 studies of 58 healthy adult subjects. The distribution of male and female subjects was 14 (47%) and 16 (53%), respectively, in the single- dose study and 16 (57%) and 12 (43%) in the multiple- dose study. In the single-dose study, 28 (93%) subjects were white; in the multidose study, all subjects were white. The mean (SD) ages in the 2 studies were 51 (10) and 51 (12) years, respectively; and mean (SD) body weights were 89.0 (10.8) and 81.1 (10.9) kg, respectively. PHX1149 exhibited dose-proportional increases in mean Cmax AUC0-t, and AUC0-infinity across the evaluated dose ranges. Tmax ranged from 2 to 4 hours, and t1/2 ranged from approximately 10 to 13 hours. No drug accumulation was observed. Plasma DPP4 inhibition at 24 hours was >or=50% in the multiple-dose study for doses of >or=100 mg. PHX1149 400 mg achieved approximately 90% 24-hour plasma DPP4 inhibition in the multiple-dose study. All AEs were characterized as mild, with the exception of 1 case of moderate edema, which occurred 17 days after the end of dosing in the multiple-dose study (50-mg dose group) and was considered unrelated to the study drug. Adverse events were experienced by 47% of all subjects studied in the single-dose study and 93% of subjects in the multiple-dose study. The rates of AEs were comparable between the study and placebo groups. CONCLUSIONS: The PK parameters and PD properties of PHX1149 were suitable (eg, tl/2, DPP4 inhibition) for once-daily dosing in this group of 58 healthy subjects. All doses were well tolerated. PMID- 17919551 TI - The effect of simvastatin on erythrocyte membrane fluidity during oxidative stress induced by cardiopulmonary bypass: a randomized controlled study. AB - BACKGROUND: Abnormal erythrocyte deformability can cause severe complications during cardiopulmonary bypass (CPB) surgery, including both hemolysis and perfusion abnormalities. OBJECTIVES: The goals of this study were to evaluate changes in erythrocyte membrane fluidity and lipid peroxidation during CPB and to examine the effect of simvastatin treatment on these parameters. METHODS: Patients undergoing cardiac surgery involving CPB were selected and randomized to receive either simvastatin 40 mg/d or placebo for 3 weeks before surgery. Three blood samples were obtained at different times during surgery for analysis of erythrocyte membrane fluidity, anion permeability, and lipid peroxidation. Erythrocyte ghosts were prepared and incubated with a lipophilic fluorescent probe (diphenyl-hexatriene), and fluorescence anisotropy was evaluated by spectrophotofluorimetric assay as a measure of membrane fluidity. Anion permeability was evaluated by the specific absorption of methemoglobin (CM) at 590 and 635 nm after treatment of heparinized blood with NaNO2. The formation of thiobarbituric acid-reactive substances was evaluated as an index of lipid peroxidation. Aspartate transaminase and lactate dehydrogenase were also measured as indices of hemolysis. RESULTS: Forty patients met the inclusion criteria (20 simvastatin, 20 placebo). Their characteristics differed significantly at baseline only in terms of the lipid profile; the statin group had higher levels of high-density lipoprotein cholesterol (P = 0.01) and lower levels of low density lipoprotein cholesterol (P = 0.001) than the placebo group. CPB was found to significantly modify characteristics of the erythrocyte membrane. Compared with preoperative values, CPB induced decreases in both mean (SD) erythrocyte membrane fluidity and anion permeability (preoperative CM: 0.69 [0.02]; 24-hour postoperative CM: 0.18 [0.02]; P < 0.001) and an increase in mean (SD) membrane lipid peroxidation (preoperative malonyl dialdehyde [MDA]: 0.21 [0.01] nmol/mL; postoperative MDA: 0.10 [0.02] nmol/mL; P < 0.001). Treatment with simvastatin was associated with a significant reduction in mean (SD) membrane lipid peroxidation both preoperatively and at 24 hours postoperatively compared with placebo (preoperative MDA: 0.07 [0.01] vs 0.10 [0.02] nmol/mL, respectively; P < 0.05; postoperative MDA: 0.10 [0.04] vs 0.21 [0.01] nmol/mL; P < 0.05). In addition, statin treatment was associated with significant increases in anion permeability preoperatively and postoperatively compared with placebo (preoperative CM: 0.79 [0.01] vs 0.69 [0.02]; P < 0.01; 24-hour postoperative CM: 0.30 [0.01] vs 0.18 [0.02]; P < 0.01). CONCLUSION: The results of this study suggest that among these patients undergoing CPB surgery, use of simvastatin for 3 weeks before the surgery had significant beneficial effects on erythrocyte membrane fluidity, lipid peroxidation, and anion permeability. PMID- 17919552 TI - Effects of milrinone on contractility and cyclic adenosine monophosphate production induced by beta1- and beta2-adrenergic receptor activation in human myocardium. AB - BACKGROUND: Because milrinone is a widely used phosphodiesterase-3 (PDE3) inhibitor, it would be of interest to know whether it interacts with beta1- and beta2-adrenergic receptor (AR) agonists in human myocardium. OBJECTIVES: This in vitro study was conducted to test whether milrinone differentially regulates cyclic adenosine-3',5'-monophosphate (cAMP) production and to examine the effect of milrinone on the positive inotropic responses and cAMP production induced by activation of the beta1-AR with norepinephrine (NE) and activation of the beta2 AR with epinephrine (EPI) in human atrial myocardium. METHODS: Right atrial trabeculae were obtained from patients undergoing cardiac surgery for valve repair. Concentration-response curves for inotropic responses mediated through the beta1-AR (NE in the presence of the beta2-blocker ICI 118, 551) and the beta2 AR (EPI in the presence of the beta1-blocker CGP 20712A) were obtained in the absence and presence of milrinone 1 micromol/L. This concentration of milrinone was chosen because it corresponded to its 50% inhibitory concentration as a PDE3 inhibitor and its therapeutic plasma concentration. The production of cAMP induced by exposure to selective beta1- and beta2-AR stimulation was also measured in the absence and presence of milrinone. RESULTS: Right atrial tissue samples were obtained from 12 white patients (7 women, 5 men; mean [SE] age, 64.6 [6.3] years) undergoing cardiac surgery for valve repair (8 mitral, 4 aortic). The presence of milrinone was associated with leftward shifts in the concentration-response curves for both NE and EPI. cAMP production in myocardial tissue samples in the presence of milrinone was increased only with NE induction (mean [SEM], 745.0 [136.7] pmol/g in the absence of milrinone vs 1620.5 [372.3] pmol/g in the presence of milrinone; P < 0.05). CONCLUSIONS: In this preliminary study in human atrial myocardium, milrinone potentiated the contractile responses to both NE and EPI. However, only the effect of NE on tissue levels of cAMP was increased in the presence of milrinone. PMID- 17919553 TI - Acute hepatitis attack after exposure to telithromycin. AB - INTRODUCTION: Antibiotic-associated hepatotoxicity is rare. With widespread use of antimicrobial agents, however, hepatic injury occurs frequently, and among adverse drug reactions, idiosyncratic reactions are the most serious. CASE SUMMARY: A 25-year-old male patient, with a height of 175 cm and weight of 72 kg presented to Marmara University Hospital Emergency Department, Istanbul, Turkey, with 5 days' history of jaundice, malaise, nausea, and vomiting. He had been prescribed telithromycin 400 mg/d PO to treat an upper respiratory tract infection 7 days prior. Admission laboratory tests were as follows: alanine aminotransferase, 67 U/L (reference range, 10-37 U/L); aspartate aminotransferase, 98 U/L (10-40 U/L); alkaline phosphatase, 513 U/L (0-270 U/L); gamma-glutamyltransferase, 32 U/L (7-49 U/L); amylase, 46 U/L (0-220 U/L); total bilirubin, 20.1 mg/dL (0.2-1.0 mg/dL); direct bilirubin, 14.8 mg/dL (0-0.3 mg/dL); and albumin, 4.7 mg/dL (3.5-5.4 mg/dL). No toxin, alcohol, or other drugs were reported. The patient had suffered a previous episode of "acute hepatitis of unknown origin," that occurred after telithromycin usage. Both incidents occurred within a year. DISCUSSION: Telithromycin is the first of the ketolide antibacterials to receive US Food and Drug Administration approval for clinical use. It has been associated with infrequent and usually reversible severe hepatic dysfunction. Based on a score of 8 on the Naranjo adverse drug reaction probability scale, telithromycin was the probable cause of acute hepatitis in this patient, and pathological findings suggested drug-induced toxic hepatitis. Recurrence of hepatitis attack might have been avoided if the initial incident had been communicated to the attending physician who prescribed telithromycin the second time. CONCLUSION: Here we report a case of acute hepatitis probably associated with the administration of telithromycin. PMID- 17919554 TI - Interaction between warfarin and cranberry juice. AB - INTRODUCTION: Warfarin is commonly used to treat or prevent thromboembolic events. Cranberry juice has been suggested to have an interaction with warfarin. However, there have been few reported cases of warfarin-cranberry juice interaction. CASE SUMMARY: A 78-year-old, 86-kg man receiving warfarin at a total weekly dose of 45 mg for atrial fibrillation presented at the Bakersfield Healthcare Center of the VA Greater Los Angeles Healthcare System with an international normalized ratio (INR) of 6.45, having reported drinking a half gallon of cranberry/apple juice in the week prior to the elevated INR. After discontinuation of the cranberry juice, maintaining the warfarin dose for 5 days, and resuming the warfarin at a total weekly dose of 40 mg, the INR returned to the therapeutic range of 2 to 3. DISCUSSION: Possible warfarin-cranberry juice interactions have been reported in the literature. Case reports illustrate INR elevation associated with cranberry juice ingestion concomitantly with warfarin administration and may be associated with bleeding (eg, pericardial, gastrointestinal). In the present case report, cranberry juice was the most likely cause of the patient's elevated INR. The Naranjo probability scale score was 3, suggesting that there was a possible interaction between warfarin and cranberry juice, while the modified Naranjo scale score adapted for anticoagulants was 5, rating the interaction as probable. CONCLUSION: The combination of warfarin administration and cranberry juice ingestion appeared to be associated with an elevated INR without bleeding in this elderly patient. PMID- 17919555 TI - A 2-week, crossover study to investigate the effect of fluticasone furoate nasal spray on short-term growth in children with allergic rhinitis. AB - BACKGROUND: It is essential to assess potential growth effects of any newly developed corticosteroid. Fluticasone furoate is a recently approved, enhanced- affinity intranasal corticosteroid with low systemic bioavailability and proven efficacy in treating allergic rhinitis. OBJECTIVE: The aim of the current study was to assess whether treatment with fluticasone furoate nasal spray affected the short-term lower-leg growth rate in children with allergic rhinitis. METHODS: Prepubertal children with seasonal or perennial allergic rhinitis of at least 1 year's duration were included in this single-center, randomized, double-blind, placebo-controlled, crossover study. The study consisted of 4 periods, each of 2 weeks' duration screening, then 2 treatment periods separated by a washout). Study medications were fluticasone furoate nasal spray 110 microg and placebo nasal spray, both administered QD in the morning. The primary end point, lower leg growth rate (measured in millimeters per week), was assessed by knemometry. Adverse events were also assessed. RESULTS: Fifty-eight patients were randomized to the study and comprised the intent-to-treat (ITT) population (mean [SD] age, 9.1 [1.4] years; 39 males, 19 females). Five patients were excluded from the ITT group due to protocol violations; thus, 53 patients (mean [SD] age, 9.0 [1.4] years; 35 males, 18 females) comprised the growth population (a de facto per protocol group). In the growth population, the adjusted mean lower-leg growth rate was 0.40 and 0.42 mm/wk with fluticasone furoate and placebo, respectively. The difference in adjusted mean lower-leg growth rate between fluticasone furoate and placebo was -0.016 mm/wk (95% CI, -0.13 to 0.10). Fluticasone furoate was noninferior to placebo, as the lower boundary of the 95% CI was above the prespecified noninferiority margin of -0.20 mm/wk. These results were supported by the ITT analysis. Fluticasone furoate nasal spray was well tolerated and had an adverse-event profile similar to that of placebo nasal spray. Nasopharyngitis (placebo, 4; fluticasone furoate, 1) and headache (placebo, 3; fluticasone furoate, 1) were the most frequent adverse events during the treatment periods. CONCLUSION: In this study, fluticasone furoate nasal spray 110 microg QD for 2 weeks had no effect on lower-leg growth rate in these prepubertal children with allergic rhinitis. PMID- 17919556 TI - Use of a Bayesian network to predict the nighttime intraocular pressure peak from daytime measurements. AB - BACKGROUND: Intraocular pressure (IOP) fluctuation over 24 hours is an independent risk factor for glaucoma progression. Nighttime lOP measurement is not routine practice. OBJECTIVE: The aim of this study was to predict, using a Bayesian network (BN), the likelihood of a nighttime IOP peak >18 mm Hg based on daytime measurements. METHODS: A pooled analysis was conducted using a BN. Data from 3 clinical trials of adult patients with glaucoma or ocular hypertension were used. IOP values at 0800, 1200, 1600, and 2000 hours were dichotomized according to the 18-mm Hg threshold. Patients' lOPs were assessed from the pretreatment washout periods and during latanoprost or travoprost treatments. A BN was constructed to study associations between daytime and nighttime IOP values, and prostaglandin analogue IOP control adjusted for trial. The nighttime IOP peak was defined as the maximum IOP value between 2400 and 0400 hours. RESULTS: The study identified 382 daily IOP vectors (6 measures per day, every 4 hours for 24 hours; pretreatment, 208; latanoprost, 73; travoprost, 101). Based on the BN, IOP at 0800 hours was associated with IOP at 1200 hours, which was also associated with the IOP at 1600 hours. IOP at 2000 hours was predicted by the IOPs at 1200 and 1600 hours. The nighttime IOP peak was associated with IOP >18 mm Hg at 1200 and 2000 hours. The percentage of patients with controlled IOP at 1200 and 2000 hours was higher in those receiving travoprost than in those receiving latanoprost. Travoprost was also associated with an increased probability of controlling nighttime IOP values >18 mm Hg (travoprost, 76.9% 77.5% vs latanoprost, 66.7%-67.9%). Daily IOP fluctuations were not found to be associated with nighttime IOP peak. CONCLUSIONS: Daytime IOP measurements are highly intercorrelated. According to this BN, IOP at 1200 and 2000 hours are more strongly associated with the nighttime IOP peak than other IOP measurements. BN can estimate the risk of a nighttime IOP peak >18 mm Hg. Daytime IOP control was important for nighttime IOP control. These findings require validation in a clinical setting. PMID- 17919557 TI - Does statin therapy initiation increase the risk for myopathy? An observational study of 32,225 diabetic and nondiabetic patients. AB - BACKGROUND: Estimates of myopathy rates in the literature are based on adverse events reported in clinical trials, which may not be representative of the clinical practice setting. OBJECTIVE: The objective of this study was to estimate the prevalence of myopathic events, particularly myalgia, myositis, and rhabdomyolysis in a community-based practice among a cohort of subjects with or without diabetes, some of whom received statin treatment. METHODS: In this retrospective data analysis, we identified members of a health maintenance organization (HMO) who initiated statin treatment between 1997 and 2004 and classified them into 2 groups: those subjects with diabetes and those without. We matched them to an equal number of health plan members based on age group, diabetes diagnosis, and year of health plan enrollment. We defined 4 levels of myopathic events according to the American College of Cardiology, the American Heart Association, and the National Heart, Lung, and Blood Institute's definitions as follows: myalgia, mild myositis, severe myositis, and rhabdomyolysis. Subjects were observed for approximately 9 years. Prevalence rates were calculated by adjusting for known myopathic risk factors and also by utilizing Cox regression models to identify predictors of time for myopathic events. RESULTS: Of the 35,413 HMO members initially assessed for inclusion, 32,225 were identified and classified into the 2 cohorts: diabetes (n = 10,247) and nondiabetes (n = 21,978). A greater proportion of statin initiators in both the diabetes (7.9% vs 5.5%; P < 0.001) and nondiabetes cohorts (9.0% vs 3.7%; P < 0.001) experienced myopathic events. However, 95% of events were myalgia or mild myositis. The prevalence of severe myositis was 0.4 per 1000 person-years (95% CI, 0.2-0.7) and 0.8 per 1000 person-years (95% CI, 0.6-1.1) among statin initiators with or without diabetes, respectively. By comparison, rates were 0.3 (95% CI, 0.1-0.5) and 0.2 (95% CI, 0.1-0.4) per 1000 person-years among nonstatin users with or without diabetes, respectively. Rates of rhabdomyolysis were 0.1 (95% CI, 0.1-0.3) and 0.2 (95% CI, 0.1-0.5) per 1000 person-years among statin and non-statin users with diabetes, respectively, and 0.2 (95% CI, 0.1-0.4) in both groups without diabetes. CONCLUSIONS: Statin initiation was associated with an approximate doubling of the risk for any myopathic event but did not appear to be associated with an increased risk for rhabdomyolysis in these patients. Because clinically significant elevations of creatine kinase levels were rare, statins appeared to be well tolerated in diabetic and nondiabetic patients who used them. PMID- 17919558 TI - Utilization of, and adherence to, drug therapy among medicaid beneficiaries with congestive heart failure. AB - BACKGROUND: Congestive heart failure (CHF) affects 4.8 million adult Americans, particularly those aged >65 years, and has been described as a "new epidemic" due to the high annual incidence of the disease (an estimated 550,000 new cases per year). OBJECTIVES: The goal of this research was to determine the number of Medicaid beneficiaries with CHF, identify the rate of CHF drug use, estimate adherence rates, examine factors associated with CHF drug use and adherence, and explore policy implications of the research findings. METHODS: Methods used included identifying noninstitutionalized beneficiaries with >or=1 inpatient claim or >or=2 ambulatory claims with a CHF diagnosis and claims for CHF drugs using 1998 State Medicaid Research Files and 1999 Medicaid Analytic eXtract data for Arkansas, California, Indiana, and New Jersey. Patient adherence was estimated using the medication possession ratio (MPR) and days of medication persistence. Multivariate regression models were used to identify factors associated with CHF drug use and adherence. RESULTS: Overall, 84.8% of beneficiaries had claims for at least 1 CHF medication; 15.2% of beneficiaries were not using any CHF medications. Among those with a claim, the mean number of claims per month was 1.4, and 25.8% had >or=4 claims per month. Mean MPR was 71.9% and mean days of medication persistence were 24.8 per month. Persons aged <65 years, men, ethnic minorities, patients with hospital admissions for conditions other than CHF, and beneficiaries with high Chronic Illness and Disability Payment System scores were less likely to have a CHF drug claim and had lower adherence rates. CONCLUSIONS: State Medicaid agencies and Medicare prescription drug plans should consider designing targeted interventions that encourage better adherence among Medicaid beneficiaries with CHF, particularly men, those aged <65 years, ethnic minorities, and patients with poor overall health status. PMID- 17919559 TI - Diffusion into use of exenatide for glucose control in diabetes mellitus: a retrospective cohort study of a new therapy. AB - BACKGROUND: Exenatide was approved by the US Food and Drug Administration (FDA) in April 2005 as adjunctive therapy to metformin or a sulfonylurea for the treatment of type 2 diabetes mellitus (DM). OBJECTIVE: We evaluated whether use of exenatide soon after its approval was consistent with the FDA- approved indications. METHODS: We assembled a retrospective cohort of patients with DM using data from a population of employed persons and their dependents, including pharmacy claims and claims for inpatient and outpatient services, provided by i3 Innovus. The data set included patients aged between 18 and 64 years with a diagnosis of DM or a claim for a DM drug from June 1, 2004, to December 31, 2005. Laboratory data were available for a subgroup of patients tested at specific commercial laboratories from June 1, 2003, to December 31, 2005. In addition, we requested data for patients with a diagnosis of obesity, regardless of a diagnosis of DM, to assess early off-label use of this medication. Patients were categorized by DM medication use and by their first fill date for exenatide, and their clinical characteristics were described. Early use was defined as filling a prescription for exenatide in the first 3 months after its approval. For descriptive purposes, we reported the means and percentages for the variables described. RESULTS: The study included data for 206,345 individuals (mean age, 51.3 years), of whom 54.0% were male. Starting in June 2005, prescriptions for exenatide were filled by 3225 (1.6%) individuals. Fifty-three percent of early users were women. Among those who filled a prescription for exenatide, 21.9% were obese, compared with 10.9% to 15.1% of those filling prescriptions for other DM medications. The proportion of patients filling a prescription for exenatide who had not received a prescription for any other DM medication in the preceding year was 14%, suggesting that exenatide was their initial therapy. A prescription for a thiazolidinedione was filled by 29.9% of patients within 60 days of filling a prescription for exenatide. CONCLUSIONS: Soon after its approval, exenatide was frequently used as monotherapy or in combination with a thiazolidinedione, neither of which is an FDA- approved indication. The observation that those filling a prescription for exenatide had a higher prevalence of obesity than those receiving prescriptions for other therapies may reflect awareness of the weight-lowering effects of exenatide. PMID- 17919560 TI - Characterizing and predicting ultrafine particle counts in Canadian classrooms during the winter months: model development and evaluation. AB - School classrooms are potentially important micro-environments for childhood exposures owing to the large amount of time children spend in these locations. While a number of airborne contaminants may be present in schools, to date few studies have examined ultrafine particle (0.02-1 microm) (UFP) levels in classrooms. In this study, our objective was to characterize UFP counts (cm(-3)) in classrooms during the winter months and to develop a model to predict such exposures based on ambient weather conditions and outdoor UFPs, as well as classroom characteristics such as size, temperature, relative humidity, and carbon dioxide levels. In total, UFP count data were collected on 60 occasions in 37 occupied classrooms at one elementary school and one secondary school in Pembroke, Ontario. On average, outdoor UFP levels exceeded indoor measures by 8989 cm(-3) (95% confidence interval (CI): 6382, 11596), and classroom UFP counts were similar at both schools with a combined average of 5017 cm(-3) (95% CI: 4300, 5734). Of the variables examined only wind speed and outdoor UFPs were important determinants of classrooms UFP levels. Specifically, each 10 km/h increase in wind speed corresponded to an 1873 cm(-3) (95% CI: 825, 2920) decrease in classroom UFP counts, and each 10000 cm(-3) increase in outdoor UFPs corresponded to a 1550 cm(-3) (95% CI: 930, 2171) increase in classroom UFP levels. However, high correlations between these two predictors meant that the independent effects of wind speed and outdoor UFPs could not be separated in multivariable models, and only outdoor UFP counts were included in the final predictive model. To evaluate model performance, classroom UFP counts were collected for 8 days at two new schools and compared to predicted values based on outdoor UFP measures. A moderate correlation was observed between measured and predicted classroom UFP counts (r=0.63) for both schools combined, but this relationship was not valid on days in which a strong indoor UFP source (electric kitchen stove) was active in schools. In general, our findings suggest that reasonable estimates of classroom UFP counts may be obtained from outdoor UFP data but that the accuracy of such estimates are limited in the presence of indoor UFP sources. PMID- 17919561 TI - Effect of protein kinase C and Ca(2+) on p42/p44 MAPK, Pyk2, and Src activation in rat conjunctival goblet cells. AB - Conjunctival goblet cells synthesize and secrete mucins onto the ocular surface to lubricate it and protect it from bacterial infections. Mucin secretion is under neural control, and cholinergic agonists released from parasympathetic nerves are major stimuli of this secretion. The signal transduction pathways these agonists use to stimulate secretion involve activating protein kinase C (PKC) and increasing intracellular [Ca(2+)] to activate the non-receptor kinases Pyk2 and p60Src (Src) to transactivate the EGF receptor. Transactivation of the EGF receptor activates a kinase cascade culminating in the activation of p42/p44 MAPK (MAPK) and ultimately that leads to secretion of high molecular weight glycocongujates (HMWGC), including mucins. To further examine the roles of PKC and Ca(2+) in the activation of MAPK, Pyk2, and Src in mucin secretion, rat conjunctival pieces and cultured goblet cells were incubated with the PKC activator phorbol myristate acid (PMA), the cholinergic agonist carbachol, or the calcium ionophore, ionomycin for varying times. Conjunctival pieces were preincubated with PKC inhibitors 10min prior to addition of carbachol (10(-4)M) for 10min. The amount of phosphorylated (activated) MAPK, Pyk2 and Src was determined by Western blotting techniques using antibodies specific to the phosphorylated forms of each kinase. PMA significantly increased the activation of MAPK, Pyk2, and Src in a time and concentration-dependent manner. PMA stimulated MAPK activity was completely inhibited by the EGF receptor inhibitor AG1478 (10(-7)M). Carbachol-stimulated MAPK activity was inhibited by three PKC inhibitors, calphostin C, chelethyrine, and staurosporine. Ionomycin (10(-6)M) stimulated MAPK activity was inhibited 66% by AG1478 (10(-7)M). Ionomycin also significantly increased Pyk2 and Src in time dependent manner. PKC and ionomycin also activated p42/p44 MAPK, Pyk2, and Src in cultured conjunctival goblet cells. We conclude that PKC and intracellular Ca(2+) activate Pyk2 and Src and phosphorylate the EGF receptor leading to stimulation of MAPK in conjunctival goblet cells. PMID- 17919562 TI - Early breast cancer therapy and cardiovascular injury. AB - Although recent advances in curative-intent therapies are beginning to produce significant survival gains in early breast cancer, these improvements may ultimately be attenuated by increased risk of long-term cardiovascular mortality. This paper reviews emerging evidence on the cardiovascular effects of breast cancer adjuvant therapy and proposes a new entity that we have labeled the "multiple-hit" hypothesis. The evidence that lifestyle modification, especially exercise therapy, may mitigate these adverse effects is also reviewed. These issues are of considerable practical importance for cardiovascular clinicians, as identification and intervention in those at high risk for cardiovascular complications may reduce a major cause of mortality in women with early breast cancer. PMID- 17919563 TI - The PROXIMAL trial: proximal protection during saphenous vein graft intervention using the Proxis Embolic Protection System: a randomized, prospective, multicenter clinical trial. AB - OBJECTIVES: To determine if outcomes could be further improved, we investigated an embolic protection device placed proximal to the target lesion that could provide protection before lesion instrumentation, allow the use of conventional guidewires, and permit embolic protection in anatomy unfavorable for distal devices. BACKGROUND: Embolic complications during stenting of degenerated saphenous vein coronary bypass grafts are reduced, but not eliminated, by distal protection. METHODS: A total of 594 patients undergoing stenting of 639 saphenous vein grafts were prospectively randomized, using a noninferiority design, to compare 2 treatment strategies: control (distal protection whenever possible) or test (proximal protection when possible, distal when not). RESULTS: The primary composite end point of death, myocardial infarction, or target vessel revascularization at 30 days by intention to treat analysis occurred in 10.0% of control and 9.2% of test patients; difference = -0.8% (95% confidence interval [CI] -5.5% to 4.0%); p for noninferiority = 0.0061. In device specific analysis, this composite end point occurred in 11.7% of distal protection patients and 7.1% of proximal protection patients (difference = -4.6% [95% CI -9.6% to 0.3%]; p for superiority = 0.10, p for noninferiority = 0.001). Finally, in the subset of patients with lesions amenable to treatment with either proximal or distal protection devices (n = 410), the primary composite end point occurred in 12.2% of distal protection patients and 7.4% of proximal protection patients; difference = -4.7% (95% CI -10.4% to 1.0%), p for superiority = 0.14, p for noninferiority = 0.001. CONCLUSIONS: Using proximal embolic protection whenever possible during treatment of diseased saphenous vein grafts produced outcomes similar to those with distal embolic protection. PMID- 17919564 TI - Unusual CD4+CD28null T lymphocytes and recurrence of acute coronary events. AB - OBJECTIVES: We hypothesized that the expansion of unusual T lymphocytes, CD4+CD28null T cells, might represent a key pathogenetic mechanism of recurrent instability. BACKGROUND: Clinical presentation of acute coronary syndromes (ACS) is variable. Some patients have recurrent episodes of instability, despite optimal treatment, whereas others have a single acute event in their life. The CD4+CD28null T cells, with a functional profile that favors vascular injury, have recently been found both in peripheral blood and in unstable coronary plaques of patients with ACS. METHODS: Peripheral blood T cells from 120 consecutive unstable angina (UA) patients were analyzed for the distribution of T-cell subsets by flow cytometry. Patients were subgrouped according to the occurrence of prior (during the 24 months before the study enrollment) and subsequent (during the 24 months of follow-up) acute coronary events. For 51 patients, the index event was the first ever (G1); 30 patients had prior events (G2); and 39 patients had further events at follow-up (death, myocardial infarction, or UA) or both before and after the index event (G3). RESULTS: The CD4+CD28null T-cell frequency was higher in G3 than in G2 and G1 (median 9.5% [range 2.4% to 48.0%] vs. 5.1% [range 0.4% to 27.8%] and 2.3% [range 0.2% to 22.8%], respectively; p < 0.001). The expansion of these unusual T lymphocytes was higher in patients with elevated C-reactive protein levels, and it was reduced by statin therapy. On multivariate logistic regression analysis, CD4+CD28null T-cell frequency was an independent predictor of future acute coronary events (odds ratio 3.01, 95% confidence interval 1.1 to 8.25; p = 0.023). CONCLUSIONS: A perturbation of T cell repertoire is strongly associated with the recurrence of acute coronary events, conceivably playing a key pathogenetic role. PMID- 17919565 TI - T cells in coronary artery disease: different effects of different T-cell subsets. PMID- 17919566 TI - The impact of for-profit hospital status on the care and outcomes of patients with non-ST-segment elevation myocardial infarction: results from the CRUSADE Initiative. AB - OBJECTIVES: We sought to determine whether for-profit status influenced hospitals' care or outcomes among non-ST-segment elevation myocardial infarction (NSTEMI) patients. BACKGROUND: While for-profit hospitals potentially have financial incentives to selectively care for younger, healthier patients, perform highly reimbursed procedures, reduce costs by limiting access to expensive medications, and encourage shorter in-patient length of stay, there are limited data available to investigate these issues objectively. METHODS: Using data from the CRUSADE (Can Rapid risk stratification of Unstable angina patients Suppress ADverse outcomes with Early implementation of the American College of Cardiology/American Heart Association guidelines) Initiative, we investigated whether for-profit status influenced hospitals' patient case mix, care, or outcomes among 145,357 patients with NSTEMI treated between January 1, 2001, and December 31, 2005, at 532 U.S. hospitals. Impact of for-profit status on care and outcomes was analyzed overall and after adjustment for clinical and facility factors using regression modeling. RESULTS: Patients (n = 11,658) treated at 58 for-profit hospitals were of similar age and gender, but were more likely to be nonwhite (black, Asian, Hispanic, and other) and have health maintenance organization/private insurance, diabetes mellitus, congestive heart failure, hypertension, and renal insufficiency compared with 133,699 patients treated at 474 nonprofit hospitals. For-profit hospitals were less likely to use discharge beta-blockers, but all other treatments were similar including the use of interventional procedures (cardiac catheterization and revascularization procedures) compared with nonprofit centers. In-hospital length of stay and mortality were also similar by hospital type. CONCLUSIONS: We found no evidence that for-profit hospitals selectively treat less sick patients, provide less evidence-based care, limit in-hospital stays, or have patients with worse acute outcomes than nonprofit centers. PMID- 17919567 TI - 64-slice computed tomography coronary angiography in patients with high, intermediate, or low pretest probability of significant coronary artery disease. AB - OBJECTIVES: We assessed the usefulness of 64-slice computed tomography coronary angiography (CTCA) to detect or rule out coronary artery disease (CAD) in patients with various estimated pretest probabilities of CAD. BACKGROUND: The pretest probability of the presence of CAD may impact the diagnostic performance of CTCA. METHODS: Sixty-four-slice CTCA (Sensation 64, Siemens, Forchheim, Germany) was performed in 254 symptomatic patients. Patients with heart rates > or =65 beats/min received beta-blockers before CTCA. The pretest probability for significant CAD was estimated by type of chest discomfort, age, gender, and traditional risk factors and defined as high (> or =71%), intermediate (31% to 70%), and low (< or =30%). Significant CAD was defined as the presence of at least 1 > or =50% coronary stenosis on quantitative coronary angiography, which was the standard of reference. No coronary segments were excluded from analysis. RESULTS: The estimated pretest probability of CAD in the high (n = 105), intermediate (n = 83), and low (n = 66) groups was 87%, 53%, and 13%, respectively. The diagnostic performance of the computed tomography (CT) scan was different in the 3 subgroups. The estimated post-test probability of the presence of significant CAD after a negative CT scan was 17%, 0%, and 0% and after a positive CT scan was 96%, 88%, and 68%, respectively. CONCLUSIONS: Computed tomography coronary angiography is useful in symptomatic patients with a low or intermediate estimated pretest probability of having significant CAD, and a negative CT scan reliably rules out the presence of significant CAD. Computed tomography coronary angiography does not provide additional relevant diagnostic information in symptomatic patients with a high estimated pretest probability of CAD. PMID- 17919569 TI - Diminished left ventricular dyssynchrony and impact of resynchronization in failing hearts with right versus left bundle branch block. AB - OBJECTIVES: We compared mechanical dyssynchrony and the impact of cardiac resynchronization therapy (CRT) in failing hearts with a pure right (RBBB) versus left bundle branch block (LBBB). BACKGROUND: Cardiac resynchronization therapy is effective for treating failing hearts with conduction delay and discoordinate contraction. Most data pertain to LBBB delays. With RBBB, the lateral wall contracts early so that biventricular (BiV) pre-excitation may not be needed. Furthermore, the magnitude of dyssynchrony and impact of CRT in pure RBBB versus LBBB remains largely unknown. METHODS: Dogs with tachypacing-induced heart failure combined with right or left bundle branch radiofrequency ablation were studied. Basal dyssynchrony and effects of single and BiV CRT on left ventricular (LV) function were assessed by pressure-volume catheter and tagged magnetic resonance imaging, respectively. RESULTS: Left bundle branch block and RBBB induced similar QRS widening, and LV function (ejection fraction, maximum time derivative of LV pressure [dP/dt(max)]) was similarly depressed in failing hearts with both conduction delays. Despite this, mechanical dyssynchrony was less in RBBB (circumferential uniformity ratio estimate [CURE] index: 0.80 +/- 0.03 vs. 0.58 +/- 0.09 for LBBB, p < 0.04; CURE 0-->1 is dyssynchronous-->synchronous). Cardiac resynchronization therapy had correspondingly less effect on hearts with RBBB than those with LBBB (i.e., 5.5 +/- 1.1% vs. 29.5 +/- 5.0% increase in dP/dt(max), p < 0.005), despite similar baselines. Furthermore, right ventricular only pacing enhanced function and synchrony in RBBB as well or better than did BiV, whereas LV-only pacing worsened function. CONCLUSIONS: Less mechanical dyssynchrony is induced by RBBB than LBBB in failing hearts, and the corresponding impact of CRT on the former is reduced. Right ventricular-only pacing may be equally efficacious as BiV CRT in hearts with pure right bundle branch conduction delay. PMID- 17919568 TI - Combined longitudinal and radial dyssynchrony predicts ventricular response after resynchronization therapy. AB - OBJECTIVES: The purpose of this study was to test the hypothesis that a combined echocardiographic assessment of longitudinal dyssynchrony by tissue Doppler imaging (TDI) and radial dyssynchrony by speckle-tracking strain may predict left ventricular (LV) functional response to cardiac resynchronization therapy (CRT). BACKGROUND: Mechanical LV dyssynchrony is associated with response to CRT; however, complex patterns may exist. METHODS: We studied 190 heart failure patients (ejection fraction [EF] 23 +/- 6%, QRS duration 168 +/- 27 ms) before and after CRT. Longitudinal dyssynchrony was assessed by color TDI for time to peak velocity (2 sites in all and 12 sites in a subgroup of 67). Radial dyssynchrony was assessed by speckle-tracking radial strain. The LV response was defined as > or =15% increase in EF. RESULTS: One hundred seventy-six patients (93%) had technically sufficient baseline and follow-up data available. Overall, 34% were EF nonresponders at 6 +/- 3 months after CRT. When both longitudinal dyssynchrony by 2-site TDI (> or =60 ms) and radial dyssynchrony (> or =130 ms) were positive, 95% of patients had an EF response; when both were negative, 21% had an EF response (p < 0.001 vs. both positive). The EF response rate was lowest (10%) when dyssynchrony was negative using 12-site TDI and radial strain (p < 0.001 vs. both positive). When either longitudinal or radial dyssynchrony was positive (but not both), 59% had an EF response. Combined longitudinal and radial dyssynchrony predicted EF response with 88% sensitivity and 80% specificity, which was significantly better than either technique alone (p < 0.0001). CONCLUSIONS: Combined patterns of longitudinal and radial dyssynchrony can be predictive of LV functional response after CRT. PMID- 17919570 TI - Restrictive right ventricular physiology: its presence and symptomatic contribution in patients with pulmonary valvular stenosis. AB - OBJECTIVES: The aim of this study was to examine whether restrictive right ventricular (RV) physiology (the presence of antegrade pulmonary arterial flow in late diastole) occurred in patients with moderate to severe isolated pulmonary valvular stenosis (PVS) and to estimate its prevalence and relationship to RV function and patient symptoms. BACKGROUND: Little is published about RV diastolic performance in adult patients with PVS. METHODS: A total of 43 consecutive patients (age 44 +/- 10 years) with moderate to severe PVS referred to Royal Brompton Hospital from 2002 to 2005 were retrospectively studied. Patient New York Heart Association (NYHA) functional class was recorded. The RV (lateral tricuspid annulus motion) long-axis movement was measured by M-mode and pulsed wave (PW) tissue Doppler imaging (TDI). Restrictive RV physiology was assessed by PW Doppler echocardiography. RESULTS: Eighteen patients (42%) had restrictive RV physiology. They were more symptomatic (NYHA functional class 1.8 +/- 0.5 vs. 1.3 +/- 0.5; p < 0.001) and had poorer RV long-axis function (TDI peak systolic velocity 7.3 +/- 2.1 cm/s vs. 9.7 +/- 2.7 cm/s; TDI early diastolic velocity 6.6 +/- 1.6 cm/s vs. 8.5 +/- 2.4 cm/s; RV long-axis systolic amplitude 1.3 +/- 0.2 cm vs. 1.5 +/- 0.3 cm; p < 0.01 for all) compared with other PVS patients despite similar RV ejection fraction, myocardial performance index, and RV systolic pressure. The presence of restrictive RV physiology (odds ratio [OR] 6.05, 95% confidence interval [CI] 1.45 to 10.29; p = 0.01) and peak pulmonary valve pressure gradient (OR 1.07, 95% CI 1.01 to 1.13; p = 0.04) were the 2 independent echocardiographic predictors for decreased exercise tolerance in patients on multivariate analysis. CONCLUSIONS: Restrictive RV physiology is common in PVS patients. Its presence is related to a worse deterioration in RV long-axis function and decreased exercise tolerance in patients. PMID- 17919571 TI - Risk factors for reoperation after repair of discrete subaortic stenosis in children. AB - OBJECTIVES: This study aimed to identify independent predictors of reoperation after successful resection of discrete subaortic stenosis (DSS). BACKGROUND: Recurrence of DSS has been reported to range from 0% to 55% of patients. Factors associated with recurrence have not been adequately defined. METHODS: Patients were included if they had a diagnosis of DSS, normal segmental cardiac anatomy, previous resection of DSS, and at least 36 months' follow-up. Demographic, surgical, and echocardiographic data were analyzed. Primary outcome was repeat resection of DSS in patients after successful primary resection. RESULTS: Of 111 subjects who had successful surgical resection of DSS, 16 patients (14%) required reoperation. Median follow-up time was 8.2 years. Form of DSS and gender did not differ significantly between those with reoperation and those without. In multivariate analysis, independent predictors of reoperation that would be available before first surgery were <6 mm distance between the aortic valve (AoV) and the obstruction (hazard ratio [HR] 5.1; p = 0.013) and peak gradient by Doppler > or =60 mm Hg (HR 4.2; p = 0.016). If intraoperative variables are also considered, peeling of the membrane from the AoV or mitral valve at first surgery, <6 mm distance between the DSS and AoV, and peak gradient by Doppler > or =60 mm Hg were independent predictors of reoperation. CONCLUSIONS: Proximity of the obstructive lesion to the AoV and severe obstruction determined by preoperative echocardiography, as well as involvement of valve leaflets requiring surgical peeling, predict recurrent DSS requiring reoperation. PMID- 17919572 TI - Reduced incidence of vagally induced atrial fibrillation and expression levels of connexins by n-3 polyunsaturated fatty acids in dogs. AB - OBJECTIVES: This open-label canine study assessed whether n-3 polyunsaturated fatty acids (PUFAs) prevent vagally induced atrial fibrillation (AF) and influence atrial tissue expression levels of connexins (CXs). BACKGROUND: n-3 polyunsaturated fatty acids in fish oils protect against sudden cardiac death and reduce postoperative AF. Changes in spatial organization of gap junctions or cellular CX levels have been linked to arrhythmogenesis. METHODS: Vagally induced AF was studied. Eight dogs were given fish oil daily for 14 days. Eight control dogs had reproducibly induced AF and were re-evaluated after intravenous administration of fish oil. Atrial fibrillation was compared, and n-3 PUFA, CX40, and CX43 protein levels were assessed in atrial biopsies. RESULTS: Atrial tissue n-3 PUFA levels increased in oral treatment dogs (5.78 +/- 0.71% vs. 2.49 +/- 0.46% in control animals, p < 0.001). No difference was observed for atrial refractory periods or hemodynamic or electrocardiographic parameters. Incidence of AF in oral treatment dogs decreased 79% with the extra stimulus technique (10.5% vs. 48.9%, p = 0.003) and 42% with burst induction (22.5% vs. 38.8%, p = 0.038). Both CX40 and CX43 levels were lower in oral treatment dogs (60% [p = 0.019] and 42% [p = 0.038] lower, respectively); protection against AF was mostly related to reduced CX40 expression levels (p = 0.02). In dogs that were given intravenous n-3 PUFAs, AF inducibility by the extra stimulus technique was reduced from 75.0% to 28.6% (p = 0.002). CONCLUSIONS: Oral treatment with fish oils increased atrial n-3 PUFA levels and reduced vulnerability to induction of AF in this dog model. Modulation of cardiac CX by n-3 PUFAs probably contributes to the antiarrhythmic effects of fish oils. PMID- 17919573 TI - Fish oil and prevention of atrial fibrillation. PMID- 17919574 TI - Impact of late drug-eluting stent malapposition on 3-year clinical events. PMID- 17919575 TI - Comparing hospital performance in door-to-balloon time between the Hospital Quality Alliance and the National Cardiovascular Data Registry. PMID- 17919576 TI - Throw the window out the door. PMID- 17919578 TI - Eliminating plaque angiogenesis. PMID- 17919580 TI - One hour electrical stimulation accelerates functional recovery after femoral nerve repair. AB - The clinical outcome of peripheral nerve injuries requiring surgical repair is usually poor and efficient therapies do not exist. Recent work has suggested that low-frequency electrical stimulation of the severed nerve which produces repeated discharges of the parent motoneuron perikarya positively influences axonal regeneration, even if applied once for a period of only 1 h. Here we provide the first evidence for locomotor functional benefits of such stimulation. We transected the femoral nerve of adult C57BL/6J mice proximal to the bifurcation of the quadriceps and saphenous branches and electrically stimulated the proximal nerve stump for 1 h at 20-Hz frequency prior to nerve repair with a silicone cuff. Three months later, the ability of the quadriceps muscle to extend the knee in sham-stimulated mice had recovered to 63% of the preoperative values as estimated by single-frame motion analysis. After electrical stimulation, the outcome was only slightly better (73%) but the rate of functional recovery was considerably accelerated. Near-maximum recovery was achieved 6 weeks earlier than in the control group. The beneficial effects were associated with larger motoneuron cell bodies and increased diameters of regenerated axons in the quadriceps nerve branch, but not with enhanced preferential reinnervation by motoneurons of muscle as opposed to skin. The observed acceleration of functional restoration and the positive effects on motoneurons and regenerated axons indicate the potential of a clinically feasible approach for improvement of nerve repair outcome in human patients in which delayed target reinnervation is a factor limiting recovery. PMID- 17919581 TI - Schistosome genomics and beyond: news and views. PMID- 17919582 TI - Litomosoides sigmodontis: vaccine-induced immune responses against Wolbachia surface protein can enhance the survival of filarial nematodes during primary infection. AB - Wolbachia are bacteria present within the tissues of most filarial nematodes. Filarial nematode survival is known to be affected by immune responses generated during filarial nematode infection and immune responses to Wolbachia can be found in different species harbouring filarial nematode infections, including humans. Using the rodent filarial model Litomosoides sigmodontis, we show that pre exposure to wolbachia surface protein in a Th1 context (but not in a Th2-context) enhances worm survival on subsequent challenge. This study suggests that despite abundant evidence that pro-inflammatory reactions to the endosymbiont have detrimental effects on the both the nematode and mammalian host, they may under some circumstances be beneficial to the nematode. PMID- 17919583 TI - Introduction advances in surgical technology supplement. PMID- 17919584 TI - The efficient delivery of elective orthopedic care. AB - The reality of the modern orthopedic practice is that rising fixed business costs, rising malpractice premiums, and decreasing reimbursement make efficiency not just a matter of prosperity, but in some markets, one of practice survival. As physicians, we have been trained to deliver medical advice to our patients and offer plans of care catered to the patient's specific needs, goals, and lifestyle. Terms such as practice efficiency may initially seem to conflict with the concept of optimizing patient outcomes. This article introduces some of the strategies that can be integrated into a practice model to increase volumes, productivity, and efficiency while simultaneously controlling costs and improving patient outcomes. PMID- 17919585 TI - Principles of a clean operating room environment. AB - Optimizing the operating room environment is necessary to minimize the prevalence of arthroplasty infection. Reduction of bacterial contamination in the operating room should be a primary focus of all members of the operating room team. However, in recent years, there has been a decline in the emphasis of the basic principles of antisepsis in many operating rooms. The purpose of this review is to highlight important considerations for optimizing the operating room environment. These principles should be actively promoted by orthopedic surgeons in their operating rooms as part of a comprehensive approach to minimizing arthroplasty infection. PMID- 17919587 TI - Knee bearing technology: where is technology taking us? AB - A novel sequentially irradiated and annealed bearing material (X3), characterized for use in knee arthroplasty, has been developed. Attention was directed to mechanical strength properties, oxidation resistance, and the ability to reduce wear. Material properties such as ultimate tensile and yield strength were unaffected by the sequential cross-linking process. Elongation was reduced relative to GUR 1020 conventional polyethylene, but equivalent to that of direct compression molded 1900 material. In knee simulator testing for normal gait and stair climbing, measured wear rates for X3 polyethylene were reduced by 79% and 77% when compared to the same knee design using conventional polyethylene. Mechanical properties and wear characteristics of the X3 polyethylene were unaffected before and after exposure to accelerated aging; properties of conventional polyethylene were adversely impacted. PMID- 17919586 TI - Pain management and accelerated rehabilitation for total hip and total knee arthroplasty. AB - Improved pain management techniques and accelerated rehabilitation programs are revolutionizing our patients' postoperative experience after total hip and knee arthroplasty. The process involves regional anesthesia with multimodal pain control using local periarticular injections in combination with enhanced patient education and accelerated rehabilitation provided by a dedicated team of surgeons, physicians, anesthesiologists, physician assistants, physical therapists, and social workers. With this system, it is now possible to achieve a painless recovery after total hip arthroplasty and total knee arthroplasty. Although this is not always the case, it was unheard of in prior years. It is our hope that future research into this area will make painful, difficult recoveries after total hip arthroplasty and total knee arthroplasty a distant memory. PMID- 17919588 TI - Minimally invasive lateral approach to total knee arthroplasty. AB - Minimally invasive knee surgery can be accomplished through several exposures used in standard total knee arthroplasty. These exposures include the medial parapatellar, subvastus, and midvastus approaches. The authors describe a new minimally invasive direct lateral approach in an attempt to minimize soft tissue damage and preserve quadriceps muscle function in 35 patients (35 knees). There were 23 women and 12 men with a mean age of 65 years. At a mean follow-up of 3.8 years, the mean Knee Society objective and functional scores improved to 94 and 92 points, respectively. The assessment of quadriceps muscle strength, anterior knee pain, and patient satisfaction was promising. However, the downside of this pilot cohort was that using instruments and implants that have not been customized for this approach led to a considerable rate of early complications that may limit the potential of this new approach. Thus, further refinements are needed to increase clinical success and allow this technique for general use. PMID- 17919589 TI - Gender-specific design in total knee arthroplasty. AB - Total knee arthroplasty (TKA) has now been performed for more than 3 decades, with millions of implantations, and a tremendous volume of scientific data regarding its use. There is little, if anything, in the academic literature to suggest an inferior result when a TKA is performed in a female patient. Recently, implant manufacturers have introduced TKA designs with modified dimensions to accommodate the anatomical differences that occur between sexes. This article will attempt to clarify the known sex issues surrounding TKA, and outline the attempts by manufacturers to improve outcomes based on sex-specific or sex friendly designs. PMID- 17919590 TI - Management of bone loss in revision total knee arthroplasty. AB - Revision total knee arthroplasty presents several complex challenges to the adult reconstructive surgeon, including management of bone loss and ligamentous insufficiency. Bone loss patterns can be anatomically categorized and the surgical treatment can be algorithmically approached based on the bone loss pattern. In this review, we present a simplified, anatomically based classification system of bone loss patterns encountered in total knee arthroplasty. This classification system can help guide the orthopedic surgeon in prosthetic design selection, and the allograft and prosthetic augmentation options in the surgical management of these cases. PMID- 17919591 TI - Hip pain in young adults: femoroacetabular impingement. AB - Femoroacetabular impingement is a more recently noticed cause of hip pain in young patients, and early diagnosis and treatment are critical to prevent development of osteoarthritis and/or delay the need for a more radical treatment option such as arthroplasty surgery. Two general types of femoroacetabular impingement have been described. In the cam impingement, the femoral deformity, usually a bump on the head-and-neck junction, impinges on the acetabular rim. The pincer type of impingement is caused by the deformity on the acetabular side, which may be a deep socket, or an acetabular overcoverage due to retroversion that creates an obstacle for flexion and internal rotation. This article attempts to examine the underlying pathogenesis and discusses diagnostic and treatment modalities. PMID- 17919592 TI - Primary total hip arthroplasty with an uncemented femoral component: two- to seven-year results. AB - This prospective study reports the midterm outcome of total hip arthroplasty performed in a consecutive series of patients using a tapered uncemented femoral component. The cohort consists of 631 patients (700 hips). The clinical records and the routine serial radiographs of these patients were monitored closely over a 5-year period. Follow-up averaged 4.35 years. There was a significant improvement in functional outcome of these patients as measured by Harris hip score and short-form 36.There were 4 revisions for aseptic loosening of the femoral component in this series, accounting for an overall survivorship of 99.4%. The study confirms that the midterm outcome of this stem is excellent, with a low revision rate. PMID- 17919594 TI - Treatment options and allograft use in revision total hip arthroplasty the acetabulum. AB - One of the most challenging aspects of revision total hip arthroplasty is management of bone loss. The acetabulum is particularly difficult, with its complex morphology and proximity to major neurovascular structures. We present a reliable classification system of pelvic and acetabular bone loss based on preoperative radiographs, and this classification system directs treatment of bone loss. The type and application of allograft bone required for the reconstruction will be reviewed for each level of bone loss. Good ability to prognosticate each level of treatment is available from follow-up studies based on survival. We know of no other scientifically developed and validated classification of pelvic bone loss that predicts the likely success of the applied treatment. PMID- 17919593 TI - The role of stripe wear in causing acoustic emissions from alumina ceramic-on ceramic bearings. AB - Noise from ceramic-on-ceramic hip bearings has been reported during specific patient activities. Wear stripes, which have been seen in some clinical retrievals, may be associated with bearing noise. We produced wear stripes on ceramic bearings in the laboratory and determined that, under certain conditions, noise could occur during either edge loading or joint simulation. Squeaking was not observed with bearings in their pristine condition. We concluded that wear stripes caused by edge loading may be associated with bearing noise, during either edge loading alone or during normal articulation. PMID- 17919596 TI - Hip resurfacing arthroplasty: the Australian experience. AB - In the last 10 years, resurfacing hip arthroplasty has become a popular option again for treating hip disorders in younger, active patients in some parts of the world. We report the Australian experience with this operation based on the Australian National Joint Replacement Registry and the literature available on the Australian experience in Resurfacing. PMID- 17919595 TI - Why total hip resurfacing. AB - Improvements in materials and fixation have addressed many of the limitations of resurfacing total hip arthroplasty (THA). The functional demands and the longevity of arthroplasty patients are increasing. Many patients have embraced the functional capacity, bone conservation, and revision options of hip resurfacing. Young men have historically been at increased risk for failure of total THA. In this patient subgroup, the survivorship of hybrid metal-metal resurfacing THA has been demonstrated to exceed that of THA, but patient selection is critical. The procedure is now also conservative on the acetabular side, and revision of the cementless acetabular component is rare. The operative parameters of femoral resurfacing revisions (conversion to a THA) are similar to that of a primary THA. Bearing technology will continue to evolve. PMID- 17919597 TI - Femoral neck fractures after metal-on-metal total hip resurfacing: a prospective cohort study. AB - There has been a renewed interest in metal-on-metal resurfacing total hip arthroplasty. Recent studies have reported high success rates at short to midterm follow-up. Despite these excellent early outcomes, femoral neck fractures have been reported as a major complication after this procedure. The purpose of this study was to identify the incidence of this complication in a prospective cohort of patients. In addition, various demographic and radiographic factors such as surgeon experience, age, sex, body mass index, femoral neck notching, and cysts were assessed as potential risk factors. Between November 2000 and August 2006, 550 metal-on-metal total hip resurfacings were performed by a single surgeon. The absolute risk for femoral neck fracture in this cohort was 2.5%. Of the 14 fractures, 12 occurred in the first 69 resurfacings performed. After this time, the incidence of fracture was 0.4%. Women and obese patients were shown to have higher cumulative incidences of fractures. These findings suggest the need for careful patient selection and surgical technique, especially for surgeons during the early learning curve for this technically difficult surgery. PMID- 17919598 TI - Whole genome microarray analysis of C. elegans rrf-3 and eri-1 mutants. AB - We performed genome wide gene expression analysis on L4 stage Caenorhabditis elegans rrf-3(pk1426) and eri-1(mg366) mutant strains to study the effects caused by loss of their encoded proteins, which are required for the accumulation of endogenous secondary siRNAs. Mutant rrf-3 and eri-1 strains exhibited 72 transcripts that were co-over-expressed and 4 transcripts co-under-expressed (>2 fold, P<0.05) compared to N2 wild type strain. Ontology analysis indicated these transcripts were over represented for protein phosphorylation and sperm function genes. These results provide additional support for the hypothesis that RRF-3 and ERI-1 act together in the endo-siRNA pathway, and furthermore suggests their involvement in additional biological processes. PMID- 17919599 TI - Oligosaccharide recognition and binding to the carbohydrate binding module of AMP activated protein kinase. AB - The AMP-activated protein kinase (AMPK) contains a carbohydrate-binding module (beta1-CBM) that is conserved from yeast to mammals. Beta1-CBM has been shown to localize AMPK to glycogen in intact cells and in vitro. Here we use Nuclear Magnetic Resonance spectroscopy to investigate oligosaccharide binding to 15N labelled beta1-CBM. We find that beta1-CBM shows greatest affinity to carbohydrates of greater than five glucose units joined via alpha,1-->4 glycosidic linkages with a single, but not multiple, glucose units in an alpha,1- >6 branch. The near identical chemical shift profile for all oligosaccharides whether cyclic or linear suggest a similar binding conformation and confirms the presence of a single carbohydrate-binding site. PMID- 17919600 TI - Macromolecular crowding increases structural content of folded proteins. AB - Here we show that increased amount of secondary structure is acquired in the folded states of two structurally-different proteins (alpha-helical VlsE and alpha/beta flavodoxin) in the presence of macromolecular crowding agents. The structural content of flavodoxin and VlsE is enhanced by 33% and 70%, respectively, in 400 mg/ml Ficoll 70 (pH 7, 20 degrees C) and correlates with higher protein-thermal stability. In the same Ficoll range, there are only small effects on the unfolded-state structures of the proteins. This is the first in vitro assessment of crowding effects on the native-state structures at physiological conditions. Our findings imply that for proteins with low intrinsic stability, the functional structures in vivo may differ from those observed in dilute buffers. PMID- 17919601 TI - Suppressive effect of reactive oxygen species on CD40-induced B cell activation. AB - Reactive oxygen species (ROS) produced by the innate immune system work as effectors to destroy pathogens and to control cellular responses. However, their role in the adaptive immune response remains unclear. Here we studied the effect of exogenous ROS on CD40-induced B cell activation. H2O2 treatment inhibited CD40 induced immunoglobulin production of B cells, DNA binding of NF-kappaB, IkappaBalpha degradation and IKK phosphorylation. On the other hand, H2O2 treatment did not induce obvious B cell death after 30 min of stimulation. Although the ligation of anti-CD40 antibody was not disturbed by H2O2, TRAF2 recruitment to CD40 was inhibited. These results suggest that exogenous ROS play a negative role in CD40 signaling during B cell activation. PMID- 17919602 TI - Wondonin, a novel compound, inhibits hypoxia-induced angiogenesis through hypoxia inducible factor 1 alpha. AB - Hypoxia regulates a variety of transcription factors including HIF-1, which has been considered a target for anti-angiogenic therapy. While searching for a novel anti-angiogenic agent that would inhibit HIF-1 activity, we identified wondonin, a new bis imidazole purified from an association of the sponges Poecillastra wondoensis. Wondonin significantly decreased hypoxia-induced HIF-1alpha protein and VEGF expression and inhibited angiogenesis in vitro and in vivo. Furthermore, wondonin down-regulated HIF-1alpha protein through the increased interaction with the pVHL in immortalized human keratinocyte cell line. Wondonin also decreased the immunoreactivities of CD31 and VEGF in epidermal hyperplasia in mice. Taken together, these results suggested that wondonin had the potential to become the anti-angiogenic therapeutic agent to target HIF-1alpha. PMID- 17919603 TI - Comparison of spindle and chromosome configuration in in vitro- and in vivo matured mouse oocytes after vitrification. AB - OBJECTIVE: To compare the cytogenetic changes in in vitro- and in vivo-matured oocytes after vitrification. DESIGN: In vitro experiments using murine model. SETTING: Animal model study in university laboratory. ANIMAL(S): CD-1 mice. INTERVENTION(S): In vitro maturation and vitrification of oocytes. MAIN OUTCOME MEASURE(S): Post-warming survival, analysis of spindle and chromosome configurations, aneuploidy screening of parthenogenetically activated oocytes, extent of DNA fragmentation, and early embryonic development after IVF. RESULT(S): Eighty percent of germinal vesicle-stage oocytes matured after in vitro maturation and were cryopreserved by vitrification (n = 354). There was no significant difference in the post-warming survival of in vitro- and in vivo matured oocytes (94.1% vs. 91.8%, respectively). The majority of in vitro- and in vivo-matured oocytes maintained normal meiotic spindle morphology and chromosome alignment (88.2% vs. 86.9%, respectively) after vitrification and the incidence of aneuploidy was not increased (11.5% vs. 9.3%). However, in vitro-matured oocytes showed a higher rate of DNA fragmentation after vitrification compared to in vivo-matured oocytes. After vitrification, the cleavage and blastocyst formation rates of in vitro-matured oocytes were significantly lower than those of in vivo-matured oocytes (37.0% vs. 60.0% and 5.4% vs. 18.9%, respectively). CONCLUSION(S): Vitrification of in vitro-matured mouse oocytes results in high survival rates, normal meiotic spindle and chromosome alignment, and no increased incidence of aneuploidy. A possible cause of the reduced developmental competence of in vitro-matured and vitrified oocytes may be due to DNA fragmentation. PMID- 17919604 TI - A child of "hers": older single mothers and their children conceived through IVF with both egg and sperm donation. AB - OBJECTIVE: To study the decision to have a child alone, the experience of gamete donation, the issue of disclosure of the donor link to the child, conception related health and sociodemographic characteristics of the mothers, children's socioemotional development, and mother-child relationships. DESIGN: Qualitative study. SETTING: Prior clients of a sperm bank. PARTICIPANT(S): Eleven single women in their late forties who gave birth to children aided by IVF involving both egg and sperm donation, i.e., the children are not genetically related to the mothers. RESULT(S): Not only were there differences among the participants, but they also differed from the only previous study focusing on single women becoming mothers by choice and using advanced reproductive technologies. CONCLUSION(S): Similarly to previous studies, we generally found that the impact of assisted conception on parenting and child development gives no undue cause for concern while the children are still young. However, the young age of the children in our sample prevented us from answering many questions about the children's socioemotional development and about disclosure of donor conception to children born to older single women using double gamete donation and IVF. PMID- 17919605 TI - Influence of heating car seats on scrotal temperature. AB - OBJECTIVE: To evaluate the influence of a heated versus an unheated car seat on scrotal temperature under standardized experimental conditions. DESIGN: Controlled clinical study. SETTING: Healthy volunteers in an academic research environment. PATIENT(S): Thirty volunteers without a history of infertility and with a normal andrological examination. INTERVENTION(S): Scrotal temperatures were measured every minute with a portable data recorder connected to two thermistor temperature sensors, which were attached on either side of the scrotum. All volunteers started the experiment at the same time of day wearing standardized cotton wool trousers and shirts fitting to body size. Each volunteer performed two periods of 90 minutes in a randomized manner on either the heated or unheated car seat. RESULT(S): At the end of the sitting periods scrotal temperatures were significantly higher using the heated car seat versus the unheated seat (left scrotal side: 0.5 degrees C; right scrotal side: 0.6 degrees C). Maximum values recorded during sitting alone were exceeded on the heated seat already after one-third of the exposure time. CONCLUSION(S): The present study suggests that the frequent use of a heated car seat represents an additional scrotal, and consequently, testicular heat stress factor to that which is present by merely sitting for long periods. PMID- 17919606 TI - Free microvascular transfer of the vermiform appendix and colon for creation of a uterovaginal fistula: a new technique for cervicovaginal reconstruction. AB - OBJECTIVE: To report a new technique for creating a uterovaginal fistula in a patient with vaginal dysgenesis, an atretic cervix, and a functional endometrium. DESIGN: Case report. SETTING: University tertiary care hospital. PATIENT(S): A patient with an atretic cervix, dysplastic vagina, and a functional uterus. INTERVENTION(S): Creation of a neocervix by partial resection of the septum within a didelphic uterus and an uterovaginal fistula by the free microvascular transfer of a vermiform appendix and an 8-cm segment of ascending colon. MAIN OUTCOME MEASURE(S): Clinical follow-up examination to verify restoration of the outflow of menstrual blood and patency of the uterovaginal fistula. RESULT(S): The patient commenced menstrual bleeding 8 weeks postoperatively, after which she had regular menstrual cycles. The uterovaginal fistula was patent at the 3-year follow-up examination. CONCLUSION(S): Free microvascular transfer of the appendix and ascending colon for creation of an uterovaginal fistula is an improved and superior technique for cervicovaginal reconstruction, with excellent long-term results. PMID- 17919607 TI - Expression of androgen receptor co-regulators in the testes of men with azoospermia. AB - OBJECTIVE: To elucidate the physiologic and pathologic roles of androgen receptor (AR) and co-regulators in human testes with obstructive azoospermia or nonobstructive azoospermia. DESIGN: Prospective laboratory and clinical study. SETTING: Infertility clinic at Chang Gung Memorial Hospital. PATIENT(S): Twenty seven men with obstructive azoospermia and 24 men with nonobstructive azoospermia. INTERVENTIONS: None. MAIN OUTCOME MEASURE(S): Expression of AR and AR co-regulators was determined in testicular specimens and were analyzed using reverse transcription polymerase chain reaction (PCR) and immunohistochemical staining techniques. RESULT(S): Most of the AR co-regulators were expressed at similar levels in specimens obtained from men with nonobstructive and obstructive azoospermia. However, the levels of expression of ARA54 mRNA were significantly lower and ARA55 mRNA was significantly higher in specimens from men with nonobstructive than obstructive azoospermia. In specimens from men with obstructive and nonobstructive azoospermia, AR immunostaining was detected in Sertoli, Leydig, and peritubular myoid cells. ARA55 immunostaining was detected in peritubular myoid and endothelial cells of blood vessels. Interestingly, nuclear ARA54 immunostaining was detected in the late stage germ cells of specimens from men with obstructive azoospermia but in the somatic cells of specimens from men with nonobstructive azoospermia. CONCLUSION(S): These results demonstrated that the decreased expression of ARA54 and increased expression of ARA55 is a feature of nonobstructive azoospermia. In addition, the differential localization of ARA54 may play an important role in testicular development and spermatogenesis in humans. PMID- 17919608 TI - Serum antimullerian hormone levels remain stable throughout the menstrual cycle and after oral or vaginal administration of synthetic sex steroids. AB - OBJECTIVE: To investigate whether oral or vaginal administration of contraceptive hormones might affect antimullerian hormone (AMH) levels. DESIGN: Prospective trial with women recruited by advertisement. Women who wished contraception were randomized between oral or vaginal estroprogestative contraception, and those who did not choose contraception were included in the control group. SETTING: Fertility clinic of a tertiary university hospital. PATIENT(S): Twenty-four young, healthy volunteer women with regular cycles who had received no hormonal contraception for at least 3 months before the study. INTERVENTION(S): Oral or vaginal estroprogestative contraception from day 5 to 25 of a menstrual cycle versus no contraception. MAIN OUTCOME MEASURE(S): Intercycle and intracycle variations of serum AMH levels in normally ovulating volunteers and following the initiation of oral or vaginal estroprogestative contraception. RESULT(S): Fluctuations of AMH levels observed during the menstrual cycle remained within cycle-to-cycle variability in cycling controls and in women receiving oral or vaginal contraception. CONCLUSION(S): Our findings confirm that AMH levels remain steady during the menstrual cycle and indicate that they are unaffected by exogenous sex steroids used for contraception whether administered orally or vaginally. PMID- 17919609 TI - Adenomyosis and endometriosis in the California Teachers Study. AB - OBJECTIVE: To evaluate the reproductive and lifestyle correlates of a surgically confirmed diagnosis of endometriosis or adenomyosis in a large prospective cohort. DESIGN: Collection of surgical diagnoses of endometriosis and adenomyosis during follow-up of women with no prior history of endometriosis and no prior surgery for adenomyosis. SETTING: The California Teachers Study (CTS), an ongoing prospective study of female teachers and school administrators established from the rolls of the California State Teachers Retirement System. PATIENT(S): Women with surgical diagnoses of endometriosis and adenomyosis were identified from California statewide hospital patient discharge records for CTS cohort members with an intact uterus and no prior history of endometriosis. Women with an incident surgical diagnosis of endometriosis (n = 229) or adenomyosis (n = 961) were compared with disease-free women in the same age range (for endometriosis, n = 43,493; for adenomyosis, n = 79,495). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Multivariable logistic regression methods were used to calculate prevalence odds ratios and associated 95% confidence intervals for the associations between self-reported menstrual and reproductive characteristics and either endometriosis or adenomyosis. RESULT(S): Women surgically diagnosed with endometriosis were younger than those surgically diagnosed with adenomyosis. Factors statistically significantly associated with endometriosis were having a mother or sister with endometriosis and nulligravidity. Factors statistically significantly associated with adenomyosis were increasing parity, early menarche (or=2 showed a trend toward association with CMV disease (hazards ratio: 2.46; 95% confidence interval: 0.66 to 9.2; p = 0.18). During the mean (+/-SD) follow-up period of 2.9 (+/-1.68) years, 3 patients died. CMV disease was not associated with all-cause mortality. CONCLUSIONS: In the contemporary era of anti viral prophylaxis, delayed-onset primary CMV disease remains a common complication among CMV D(+)/R(-) cardiac recipients. This finding warrants a better strategy for CMV prevention. PMID- 17919623 TI - The utility of right ventricular endomyocardial biopsy for the diagnosis of xenograft rejection after CD46 pig-to-baboon cardiac transplantation. AB - INTRODUCTION: Endomyocardial biopsy is the standard means of establishing cardiac allograft rejection diagnosis. The efficacy of this procedure in xenotransplantation has not been determined. In this study we compare the histology of right ventricular endomyocardial biopsy specimens with the corresponding full cross sections of explanted right ventricle (RV). We also compare RV with the related left ventricle (LV) cross sections. METHODS: Heterotopic CD46 pig-to-baboon cardiac xenotransplants (n = 64) were studied. RV endomyocardial biopsy specimens were taken at cardiac explant by using a standard bioptome (n = 24) or by sharp dissection (n = 40). Hematoxylin and eosin stained sections of RV and LV cross-section and RV endomyocardial biopsy specimens were compared in a blinded fashion. Characteristics of delayed xenograft rejection and a global assessment of ischemia were scored from 0 to 4 according to the percentage of myocardium involved (0, 0%; 1, 1%-25%; 2, 26%-50%; 3, 51%-75%; and 4, 76%-100%). RESULTS: Median graft survival was 30 days (range, 3-137 days). Linear regression analysis of histology scores demonstrated that specimens from both bioptome and sharp dissection equally represented the histology of the RV cross section. Global ischemic injury was strongly correlated between RV and RV endomyocardial biopsy (R(2) = 0.84) and between RV and LV cross sections (R(2) = 0.84). Individual characteristics of delayed xenograft rejection showed no significant variation between RV and RV endomyocardial biopsy or between RV and LV (p < 0.05). CONCLUSIONS: These results indicate that delayed xenograft rejection is a widespread process involving both right and left ventricles similarly. This study shows that histologic assessment of RV endomyocardial biopsy specimens is an effective method for the monitoring of delayed xenograft rejection after cardiac xenotransplantation. PMID- 17919625 TI - The endothelin axis and gelatinase activity in alveolar macrophages after brain stem death injury: a pilot study. AB - BACKGROUND: Endothelin-1 (ET-1) is a potent vasoconstricting mitogen that has been implicated in the development of primary graft dysfunction. Increased activity of matrix metalloproteinases (MMPs), specifically MMP-2 and -9, has been associated with tissue damage in acute lung injury and after lung transplantation. Using a validated model of brain-stem death (BSD), we aimed to determine whether alveolar macrophage up-regulation in the pulmonary system is an early feature of BSD injury and if expression levels of ET-1, endothelin A receptors (ET(A)R) and endothelin B receptors (ET(B)R), as well as MMP-2 and -9, are increased in comparison to sham controls. METHODS: Six control and 8 experimental Wistar-Kyoto rats had a balloon catheter inserted into their subdural space. In the experimental group the balloon was inflated for 4 hours. Lung specimens were immunohistochemically labeled with CD68, ET-1, ET(A)R, ET(B)R, MMP-2 and MMP-9, and 10 fields per slide were assessed. RESULTS: The ratio of alveolar macrophages to polymorphonuclear neutrophils was significantly greater in the BSD group than in controls (9 +/- 4.1 vs 3 +/- 0.5, p = 0.004) and adventitial macrophages increased in BSD lung parenchyma (p < 0.0001). ET-1, ET(A)R and ET(B)R levels were elevated in the experimental group (27.6 +/- 5.7 vs 7 +/- 2.3, 36.1 +/- 4.6 vs 17.7 +/- 2.6 and 60 +/- 7.1 vs 19.8 +/- 3.7, p < 0.0001 inclusive). BSD expression of MMP-2 and MMP-9 was double that of controls (14.9 +/- 3.4 vs 30.7 +/- 3.4 and 14.2 +/- 2.2 vs 37 +/- 3.6, respectively, p < 0.0001 inclusive). CONCLUSIONS: Alveolar macrophages are rapidly recruited after BSD and may affect peri-operative lung function via increased expression of ET-1, ET(A)R, ET(B)R, MMP-2 and MMP-9. PMID- 17919624 TI - Beta-blockers influence the short-term and long-term prognostic information of natriuretic peptides and catecholamines in chronic heart failure independent from specific agents. AB - INTRODUCTION: In chronic heart failure (CHF), the physiologic effects of natriuretic peptides and catecholamines are interdependent. Furthermore, reports state an agent-dependent effect of individual beta-blockers on biomarkers. Data on the short-term and long-term predictive power comparing these biomarkers as well as accounting for the influence of beta-blocker treatment both on the marker or the resultant prognostic information are scarce. METHODS: We included 513 consecutive patients with systolic CHF, measured atrial natriuretic peptide (ANP), N-terminal prohormone brain natriuretic peptide (NTproBNP), noradrenaline, and adrenaline, and monitored them for 90 +/- 25 months. Death or the combination of death and cardiac transplantation at 1 year, 5 years, and overall follow-up were considered end points. RESULTS: Compared with patients not taking beta blockers, patients taking beta-blockers had significantly lower levels of catecholamines but not natriuretic peptides. Only for adrenaline was the amount of this effect related to the specific beta-blocker chosen. Receiver operating characteristic curves demonstrated superior prognostic accuracy for NTproBNP both at the 1- and 5-year follow-up compared with ANP, noradrenaline, and adrenaline. In multivariate analysis including established risk markers (New York Heart Association functional class, left ventricular ejection fraction, peak oxygen uptake, and 6-minute walk test), of all neurohumoral parameters, only NTproBNP remained an independent predictor for both end points. CONCLUSION: Long-term beta blocker therapy is associated with decreased levels of plasma catecholamines but not natriuretic peptides. This effect is independent from the actual beta-blocker chosen for natriuretic peptides and noradrenaline. In multivariate analysis, both for short-term and long-term prediction of mortality or the combined end point of death and cardiac transplantation, only NTproBNP remained independent from established clinical risk markers. PMID- 17919626 TI - Donor cardiac troponin I levels do not predict recipient survival after cardiac transplantation. AB - BACKGROUND: Serum levels of cardiac troponin I (cTnI) are frequently measured in the evaluation of potential heart donors. However, the utility of cTnI levels for predicting recipient outcomes remains controversial. This study was performed to determine whether donor cardiac cTnI levels exceeding 1.0 microg/liter are associated with adverse recipient outcomes. METHODS: All donors managed by the California Transplant Donor Network between January 2001 and July 2002 with consent for donor evaluation and at least 1 measured cTnI level were included in the study if 1-year recipient mortality data were available. Each study subject was classified as having elevated cTnI if any level exceeded 1.0 microg/liter. Donor variables, recipient risk of 30-day and 1-year mortality, and recipient need for mechanical circulatory support were compared between the 2 groups. RESULTS: A total of 263 potential donors were evaluated, and 98 had elevated cTnI levels. Of these potential donors, 139 were accepted for transplantation. The cTnI levels were normal in 96 and elevated in 43. Most donors (77%) with elevated cTnI levels had levels of less than 10 microg/liter. Donor cardiopulmonary resuscitation was associated with cTnI elevations. Donors with elevated cTnI levels did not require higher doses of inotropic drugs before transplantation and had similar hemodynamic profiles compared with donors with normal cTnI levels. Although there was a trend towards longer post-transplant hospitalization in recipients of grafts from donors with elevated cTnI levels (17 days vs 15 days, p = 0.044), there was no significant difference in the recipient need for mechanical circulatory support or 30-day and 1-year mortality between the 2 groups. CONCLUSIONS: In this study, a modestly elevated donor cTnI was not associated with a higher risk of recipient mortality or need for post-transplant mechanical circulatory support. A potential donor heart should not be discarded solely because the troponin level is elevated. PMID- 17919627 TI - Fibrinolytic treatment improves the quality of lungs retrieved from non-heart beating donors. AB - BACKGROUND: The use of non-heart-beating donors (NHBDs) is an alternative strategy to increase the limited number of donors. The ex vivo evaluation has been proposed to assess the function of the lungs from NHBDs as an interim evaluation of the graft before transplantation. We evaluated the effect of a fibrinolytic agent, urokinase, in a pig ex vivo evaluation model. METHODS: Domestic pigs (30-38 kg) were divided in 3 groups of 5 pigs each. In the Control Heart-Beating Donor (HBD) Group, the lungs were flushed, explanted, and stored in cold solution (4 degrees C) of low potassium dextran for 4 hours. The pigs in the other 2 study groups were non-heart-beating donors (NHBD), and their lungs were topically cooled for 1 hour in the closed chest after 3 hours of warm ischemia. Urokinase (100,000 IU) was added into the perfusate during reperfusion 1n 1 of the NHBD groups (NHBD-UROK). Hemodynamic and aerodynamic parameters were measured. The wet-to-dry weight ratio was calculated. RESULTS: There was a significant difference between NHBD-UROK and NHBD Groups in pulmonary vascular resistance (22.5 +/- 3.06 vs 39.02 +/- 6.6 Wood Units, p = 0.032), partial pressure of arterial oxygen/fraction of inspired oxygen (250.8 +/- 23.3 vs 148.9 +/- 14.6 mm Hg, p = 0.032), oxygenation index (6.9 +/- 0.7 vs 15.9 +/- 3.2, p = 0.016), and wet-to-dry weight ratio (5.99 +/- 0.2 vs 7.74 +/- 0.3, p = 0.016). Pulmonary vascular resistance did not differ between the HBD and NHBD-UROK Groups but was significantly higher in the NHBD Group than in the HBD Group (p = 0.032). CONCLUSION: Adding urokinase into the perfusate during ex vivo evaluation resulted in improved graft function by reducing pulmonary vascular resistance and increasing oxygenation after 3 hours of warm ischemia. This ex vivo evaluation model is feasible and may be used to recondition grafts from NHBDs. PMID- 17919628 TI - Treatment of unprotected left main coronary artery stenosis in a 5-year-old heart transplant patient using a sirolimus-eluting stent. AB - Cardiac allograft vasculopathy is the most common cause of long-term graft failure in adult and pediatric heart transplant recipients. In the absence of a specific treatment for this condition, percutaneous revascularization has been the main palliative treatment in the adult population. Revascularization of pediatric patients, however, is more problematic secondary to the lack of a large pool of outcome data and the encounter of special technical challenges. We present the case of a 5-year-old girl who presented with severe cardiac allograft vasculopathy of her left main coronary artery and was treated with sirolimus stent placement. PMID- 17919629 TI - Stroke and myocardial infarction as late complications of lung transplantation. AB - A 64-year-old man who had received a lung transplant later presented with an air embolism that caused ST-segment elevation myocardial infarction, multiple strokes, and death. Transesophageal echocardiography was used to document air bubbles crossing from a bronchial fistula to a pulmonary vein and into the left atrium. Spontaneous air was seen entering a pulmonary vein during positive pressure ventilation and exiting through the left ventricular outflow tract. Autopsy confirmed the presence of a probe-patent bronchial-to-pulmonary vein fistula within a focus of necrosis and infection with Aspergillus flavus, an angioinvasive organism. The potential for intravascular gas arising from the anastomotic site should be considered when transplant recipients who present with myocardial or peripheral arterial infarction are evaluated. PMID- 17919630 TI - Early cardiac tamponade due to tension pneumopericardium after bilateral lung transplantation. AB - We report the case of a 42-year-old woman who developed severe hemodynamic instability with marked arterial pulsed pressure variation in the early course of bilateral lung transplantation. The diagnosis of tension pneumopericardium was made on Day 2 post-operatively based on chest X-ray and echocardiography. Transoesophageal echocardiography revealed both a cardiac tamponade and a right to-left shunt via a patent foramen ovale. The treatment and mechanisms of these two rare complications are discussed. PMID- 17919631 TI - Left pneumonectomy in a patient with a chronically infected allograft. AB - We describe a young man with cystic fibrosis who underwent bilateral sequential lung transplantation (BSLT) and a subsequent right single-lung re-transplant for bronchiolitis obliterans syndrome (BOS). Destruction of the retained left lung with recurrent pneumonia, worsening bronchiectasis and abscess formation was treated with a left lower lobectomy and a subsequent complete pneumonectomy. The patient tolerated the procedures and is alive and well 18 months after left pneumonectomy. In the setting of BOS, allograft pneumonectomy can be performed safely to remove non-functioning infected tissue. PMID- 17919632 TI - Exercise-induced increase in brain natriuretic peptide is related to vascular endothelial function after heart transplantation. PMID- 17919633 TI - Microbial mannan inhibits bacterial killing by macrophages: a possible pathogenic mechanism for Crohn's disease. AB - BACKGROUND & AIMS: Crohn's disease (CD) is mimicked by inherited phagocyte disorders and is associated with circulating antibodies against yeast mannan (anti-Saccharomyces cerevisiae antibody; ASCA). We speculated that mannans might impair phagocyte function. METHODS: S cerevisiae mannan was assessed for its effects on human peripheral blood neutrophils, adherent monocytes, and monocyte derived macrophages (MDM). RESULTS: Mannan caused dose-related increased survival of CD Escherichia coli HM605 within adherent monocytes from 24% +/- 10.5% (control) to 114% +/- 22.7% with mannan 1 mg/mL at 2 hours (mean +/- SEM, n = 9; P = .0002). Electron microscopy showed E coli HM605 surviving and probably replicating within macrophage vesicles. Mannan (1 mg/mL) inhibited the respiratory burst in neutrophils and monocytes (both P = .002) and bacterial killing within MDM (P < .001). E coli survival was increased within macrophages from TLR4(-/-) (126% +/- 3.5% survival at 2 hours) and MyD88(-/-) (134.8% +/- 6.5%) mice compared with wild-type mice (both P < .0001). Mannan had no additional effect, showing that TLR4 and MyD88 are involved in bacterial killing by macrophages and its inhibition by mannan. Putative CD-associated micro organisms were screened for the ASCA mannan epitope by Galanthus nivalis lectin (GNA) blotting. ASCA epitope was expressed by Candida albicans and Mycobacterium paratuberculosis but not by Mycobacterium tuberculosis or E coli. Supernatants from M paratuberculosis culture inhibited killing of E coli HM605 by adherent human monocytes and murine macrophages. The inhibitory activity was removed by GNA-affinity chromatography. CONCLUSIONS: Suppression of mucosal phagocyte function by microbial mannans, possibly of Mycobacterial origin, may contribute to CD pathogenesis. PMID- 17919635 TI - Literacy as the foundation of patient responsibility. PMID- 17919634 TI - Hypnotherapy for children with functional abdominal pain or irritable bowel syndrome: a randomized controlled trial. AB - BACKGROUND & AIMS: Functional abdominal pain (FAP) and irritable bowel syndrome (IBS) are highly prevalent in childhood. A substantial proportion of patients continues to experience long-lasting symptoms. Gut-directed hypnotherapy (HT) has been shown to be highly effective in the treatment of adult IBS patients. We undertook a randomized controlled trial and compared clinical effectiveness of HT with standard medical therapy (SMT) in children with FAP or IBS. METHODS: Fifty three pediatric patients, age 8-18 years, with FAP (n = 31) or IBS (n = 22), were randomized to either HT or SMT. Hypnotherapy consisted of 6 sessions over a 3 month period. Patients in the SMT group received standard medical care and 6 sessions of supportive therapy. Pain intensity, pain frequency, and associated symptoms were scored in weekly standardized abdominal pain diaries at baseline, during therapy, and 6 and 12 months after therapy. RESULTS: Pain scores decreased significantly in both groups: from baseline to 1 year follow-up, pain intensity scores decreased in the HT group from 13.5 to 1.3 and in the SMT group from 14.1 to 8.0. Pain frequency scores decreased from 13.5 to 1.1 in the HT group and from 14.4 to 9.3 in the SMT group. Hypnotherapy was highly superior, with a significantly greater reduction in pain scores compared with SMT (P < .001). At 1 year follow-up, successful treatment was accomplished in 85% of the HT group and 25% of the SMT group (P < .001). CONCLUSIONS: Gut-directed HT is highly effective in the treatment of children with longstanding FAP or IBS. PMID- 17919636 TI - Operative treatment of scapular fractures: a systematic review. AB - We systematically reviewed the published evidence regarding the operative treatment of scapular fractures. Publications were identified using MEDLINE databases and were included if they reported operative indications, surgical approach and implants, postoperative complications, and functional outcomes. Seventeen investigations encompassing 243 cases met our eligibility criteria. All were retrospective case series (evidence-based medicine level IV). The most common injuries treated with surgery were glenoid fossa fractures and scapular neck fractures. Approximately 25% of the cases had a concomitant injury to the clavicle or acromioclavicular ligaments. Internal fixation was most often achieved with a plate and screws through a posterior approach. The complication rate was low with infection, shoulder stiffness, and implant failure the most commonly reported problems. Good to excellent functional results were obtained in approximately 85% of the cases an average of 49.9 months postoperatively. PMID- 17919637 TI - Major trauma & cervical clearance radiation doses & cancer induction. AB - AIM: To compare the radiation dose of cervical spine clearance and body CT in a cohort of unconscious, major trauma patients for three different protocols, comparing spiral to multislice CT. To quantify the radiation exposure effect of the protocols on the lifetime cancer risk. METHOD: The hospital trauma database was used to find the unconscious (GCS<9), severely injured (Injury Severity Score >15) from 1 January 2001 to 31 December 2003, excluding isolated head injuries. The protocols used for imaging the brain and cervical spine were, including the radiographs performed as a mode: The exposure factors and field of view used were put into the Monte Carlo software, to estimate the CT and radiographic X-ray doses to the body as a whole and the dose to the thyroid associated with each region imaged. The associated nominal additional lifetime cancer risk was assessed. RESULTS: Excluding inter hospital transfers, where data was incomplete, 87 patients survived to be admitted and fulfilled the criteria. In 30 cases, the CT films were missing, the exposure factors were not recorded or no imaging was performed. In a further 21 cases, the X-ray packets were missing. Three patients had brain and cervico-dorsal CT imaging only, leaving 33 cases for evaluation. The effective radiation dose for a spiral CT of the brain using the Toshiba Xpress GX CT scanner was 3.8 mSv. The total effective doses for imaging the brain and cervical spine using the three protocols with the same CT scanner were (S.D. as % of mean): (1) 4.4 mSv (5%), (2) 7.1 mSv (10%) and (3) 8.2 mSv (15%). The corresponding mean thyroid doses were: (1) 8.5 mGy (25%), (2) 48.9 mGy (20%) and (3) 66.5 mSv (20%). The resultant nominal lifetime cancer risks were: (1) 1:4500, (2) 1:2800 and (3) 1:2400. For the Siemens Sensation 16 multislice CT scanner, the total effective doses (S.D. as % of mean) were: (1) 2.3 mSv (10%), (2) 4.3 mSv (25%) and (3) 5.4 mSv (35%). The mean doses to the thyroid were: (1) 5.9 mGy (30%), (2) 36.1 mGy (50%) and (3) 52.4 mGy (40%). The lifetime cancer risks were: (1) 1:8700, (2) 1:4600 and (3) 1:3700. Using the Toshiba spiral CT scanner, the total dose and additional lifetime nominal cancer risk associated with CT of the chest, abdomen and pelvis (CAP) as 16 mSv and 1:1250, respectively. Using the Siemens multislice CT scanner, these were 11.8 mSv and 1:1700. The cancer risk for protocol 1 when combined with a CT scan of the chest, abdomen and pelvis was 1:1000 for the spiral CT scanner and 1:1500 for the multislice CT (MCT) scanner. The cancer risk for protocol 2 with CAP CT using the MCT was 1:1200. The cancer risk for protocol 3 when combined with a CT scan of the chest, abdomen and pelvis was 1:1100 for the multislice CT scanner. Prior to the introduction of the BTS guidelines for cervical clearance, 12% of cases had CT of the body, which increased to 16% post-guidelines. CONCLUSIONS: CT of the trunk (chest, abdomen and pelvis) is associated with the greatest risk of inducing a fatal cancer in the severely injured patient with a GCS less than 9. In our institution the multislice CT protocols expose the patient to less radiation than single slice CT, which is contrary to much of the published work to date. CT scanning the thyroid (or whole cervical spine) still has a marked effect on the cancer risk in cervical clearance. Many centres will relax cervical spinal precautions in unconscious trauma patients if the cervical spine CT with reconstructions is normal. CT of the whole cervical spine may be justified in the unconscious, severely injured patient. In conscious trauma patients, the additional lifetime risk may not justify CT of the whole cervical spine as a routine practice. PMID- 17919639 TI - [Hemoglobin target for chronic kidney disease patients?]. PMID- 17919638 TI - Alternating pressure air mattresses as prevention for pressure ulcers: a literature review. AB - OBJECTIVES: The purpose of this paper is to examine and synthesise the literature on alternating pressure air mattresses (APAMs) as a preventive measure for pressure ulcers. DESIGN: Literature review. DATA SOURCES: PubMed, Cinahl, Central, Embase, and Medline databases were searched to identify original and relevant articles. Additional publications were retrieved from the references cited in the publications identified during the electronic database search. RESULTS: Thirty-five studies were included. Effectiveness and comfort of APAMs were the main focuses of the studies evaluating APAMs. Pressure ulcer incidence, contact interface pressure, and blood perfusion were the most frequently used outcome measures to evaluate the effectiveness of APAMs. Fifteen randomised controlled trials (RCTs) analysed the pressure ulcer incidence. One RCT compared a standard hospital mattress with an APAM and found that the APAM was a more effective preventive measure. RCTs comparing APAMs with constant-low-air mattresses resulted in conflicting evidence. There was also no clear evidence as to which type of APAM performed better. All RCTs had methodological flaws. The use of contact interface pressure and blood perfusion measurements to evaluate the effectiveness of APAMs is questionable. Comfort of APAMs was the primary outcome measure in only four studies. Different methods for assessment were used and different types of APAMs were evaluated. Better measures for comfort are needed. A few studies discussed technical problems associated with APAMs. Educating nurses in the correct use of APAMs is advisable. CONCLUSION: Taking into account the methodological issues, we can conclude that APAMs are likely to be more effective than standard hospital mattresses. Contact interface pressure and blood perfusion give only a hypothetical conclusion about APAMs' effectiveness. Additional large, high-quality RCTs are needed. No conclusions can be drawn regarding the comfort of APAMs. A number of technical problems associated with APAMs are related to nurses' improper use of the devices. PMID- 17919640 TI - [Measuring quality of life in end-stage renal disease. Transcultural adaptation and validation of the specific Kidney Disease Quality of Life questionnaire]. AB - End-stage renal disease has an important impact on the patients' daily life, which can be measured by quality of life questionnaires. The objective of this work was to adapt the Kidney Disease Quality of Life questionnaire (KDQoL) into French and to determine its basic psychometric properties, i.e. validity and reliability. The KDQoL consisted of 8 generic dimensions and 11 specific dimensions. The questionnaire was translated several times independently, and then submitted to a committee of professionals. The study of the measurement properties was carried out near 68 dialysis patients. KDQoL is valid and reproducible, and has properties comparable to the original instrument: missing items proportion of 5.5%, limited floor and ceiling effects (except for 4 dimensions), Cronbach alpha coefficient varying from 0.64 to 0.92 (except for 2 dimensions), test-retest coefficient greater than 0.67 (except for 3 dimensions), and the items of KDQoL were better correlated with their dimension than with other dimensions (except for 2 dimensions). Correlations between the generic and the specific scores showed the absence of redundancies between specific and generic dimensions. Thus the French version has comparable properties to the original KDQoL. This questionnaire can be used to measure the quality of life of the dialysis patients. It constitutes a good tool in clinical research, allowing international comparisons. PMID- 17919642 TI - [Diuretic-based therapy]. AB - Diuretics are pharmacological agents that increase natriuresis through inhibition of tubular re-absorption of sodium. The mechanisms and site of this inhibition differ with each drug class, accounting for their additive effects on natriuresis increase and their hydroelectrolytic side effects. The response to a given diuretic dose depends on the diuretic concentration on the urine at its action site. This concentration may be decreased by pharmacokinetic factors such as encountered in renal insufficiency or in nephrotic syndrome. These resistance mechanisms of diuretics may be corrected by dose increase, previous diuretic fixation on albumin or warfarin administration. Once these mechanisms are opposed, the diuretic concentration for maximal efficacy is reached at is action site and the natriuresis obtained as the normal maximal plateau. This is not the case when an oedematous systemic disease with effective hypovolemia is present, like in heart failure or cirrhosis, or when chronic use of loop diuretics has induced a hypertrophy of the more distant part of the tubule. In theses cases, a pharmacodynamic resistance exists, resulting in a lower maximal natriuresis plateau in spite of adequate concentration of the diuretic at its action site, even in the absence of pharmacokinetic resistance factors. The main indications of diuretics are systemic oedematous disease and hypertension. In the oedematous diseases, diuretics indication is both straightforward and sufficient only if effective hypervolemia is present. The therapeutic approach is discussed according to the various clinical conditions and pathophysiological background. In uncomplicated hypertension, diuretics are the cornerstone of the therapy. The most suitable diuretic treatment for hypertension is an association of low doses thiazide (12.5-50 mg/day) with potassium sparing diuretics. Rare indications of diuretics are also reviewed. PMID- 17919641 TI - [Microalbuminuria and urinary albumin excretion: clinical practice guidelines]. AB - Measurement of urinary albumin excretion (UAE) may be done on a morning urinary sample or on a 24 hours-urine sample. Values defining microalbuminuria are: 24 hour-urine sample: 30-300 mg/24 hours; morning urine sample: 20-200 mg/ml or 30 300 mg/g creatinine or 2.5-25 mg/mmol creatinine (men) or 3.5-35 mg/mol (women). Timed urine sample: 20-200 microg/min. The optimal use of semi-quantitative urine test-strip is not clearly defined. It is generally believed that microalbuminuria reflects a generalized impairment of the endothelium; however, no definite proof has been shown in humans. IN DIABETIC SUBJECTS: Microalbuminuria is a marker of increased risk of cardiovascular (CV) and renal morbidity and mortality in type 1 and type 2 diabetic subjects. The increase in UAE during follow-up is also a marker of CV and renal risk in type 1 and type 2 diabetic subjects; its decrease during follow-up is associated with lower risks. IN NO DIABETIC SUBJECTS: Microalbuminuria is a marker of increased risk for diabetes mellitus, deterioration of the renal function, CV morbidity and all-cause mortality. It is a marker of increased risk for the development of hypertension in normotensive subjects, and is associated with unfavorable outcome in patients with cancer and lymphoma. Persistence or elevation of UAE overtime is associated with deleterious outcome in some hypertensive subjects. Measurement of UAE may be recommended in hypertensive subjects with one or two CV risk factors in whom CV risk remains difficult to assess, and in those with refractory hypertension: microalbuminuria indicates a high CV risk and must lead to strict control of arterial pressure. Studies focused on microalbuminuria in non-diabetic non-hypertensive subjects are limited; most of them suggest that microalbuminuria predicts CV complications and deleterious outcome as it is in diabetic or hypertensive subjects. Subjects with a history of CV or cerebrovascular disease have an even greater CV risk if microalbuminuria is present than if it is not; however, in all cases, therapeutic intervention must be aggressive regardless of whether microalbuminuria is present or not. It is not recommended to measure UAE in non-diabetic non-hypertensive subjects in the absence of history of renal disease. Monitoring of renal function (UAE, serum creatinine and estimation of GFR) is annually recommended in all subjects with microalbuminuria. MANAGEMENT: In patients with microalbuminuria, weight reduction, sodium restriction (<6 g/day), smoking cessation, strict glucose control in diabetic subjects, strict arterial pressure control are necessary; in diabetic subjects: use of maximal doses of ACEI or ARB are recommended; ACEI/ARB and thiazides have synergistic actions on arterial pressure and reduction of UAE; in non diabetic subjects, any of the five classes of antihypertensive medications (ACEI, ARB, thiazides, calcium channel blockers or betablockers) can be used. PMID- 17919643 TI - [Biophysics of membrane transport and recent progress in hemodialysis]. PMID- 17919644 TI - Cyclophilin A differentially activates monocytes and endothelial cells: role of purity, activity, and endotoxin contamination in commercial preparations. AB - BACKGROUND: Cyclophilin A (CyPA) is a cytoplasmic protein secreted under inflammatory conditions. Extracellular CyPA is detected in atherosclerotic plaques and has been observed to activate endothelial cells as well as monocytes. METHODS AND RESULTS: Commercially available recombinant CyPA-induced expression of tissue factor (TF) and vascular cell adhesion molecule-1 (VCAM-1) in human aortic endothelial cells (HAEC). However, CyPA from commercial sources contained lipopolysaccharide at concentrations up to 18.9 ng/ml; moreover, it exhibited low purity as determined by protein spectrum analysis and low activity as assessed by peptidyl prolyl cis-trans isomerase (PPIase) assay. An in-house preparation of pure, active, and uncontaminated CyPA failed to induce endothelial TF or VCAM-1 expression; moreover, it was not chemotactic for HAEC. In contrast, such CyPA exhibited potent chemotactic activity on monocytic THP-1 cells, with a maximal effect on migration occurring at a concentration of 5.5 x 10(-9)mol/l. Pretreatment of CyPA with cyclosporine A prevented its effect on THP-1 cell migration; similarly, PPIase-deficient mutant CyPA protein did not induce migration of these cells. In-house prepared CyPA induced the release of Il-6, but not TNF-alpha, from THP-1 cells. CONCLUSIONS: Commercially available CyPA exhibits low purity and activity and may be contaminated by endotoxin. Pure, active, and uncontaminated CyPA does not induce endothelial TF or VCAM-1 expression; instead, it acts as a potent monocyte chemoattractant and induces monocyte Il-6 release, implying a role for extracellular CyPA in the pathogenesis of atherosclerosis via activation of monocytes rather than endothelial cells. PMID- 17919645 TI - Carotid geometry effects on blood flow and on risk for vascular disease. AB - It has been widely observed that atherosclerotic diseases occur at sites with complex hemodynamics, such as artery bifurcations, junctions, and regions of high curvature. These regions usually have very low or highly oscillatory wall shear stress (WSS). In the present work, 3D pulsatile blood flow through a model of the carotid artery bifurcation was simulated using a finite volume numerical method. The goal was to quantify the risk of atherogenesis associated with different carotid artery geometries. A risk scale based on the average WSS on the sinus wall of the internal carotid artery was proposed-a scale that can be used to quantify the effect of the carotid geometry on the relative risk for developing vascular disease. It was found that the bifurcation angle and the out-of-plane angle of the internal carotid artery affect the formation of low stress regions on the carotid walls. The main conclusions are: (a) larger internal carotid artery angles (theta(IC)) generally increase the frequency and the area of blood recirculation and lower the WSS on the sinus wall, hence increasing the risk of plaque build-up; (b) off-plane angles were found to lower the WSS on the sinus for geometries with theta(IC)25 degrees . Larger off-plane angles generally increase the danger of plague build-up; (c) for theta(IC) < 25 degrees , the off plane angle does not have an obvious effect on the hemodynamic WSS; (d) symmetric bifurcations were found to increase the WSS on the sinus wall and ease the risk of vascular disease. PMID- 17919646 TI - Thermodynamics of hydrophobic interaction chromatography of acetyl amino acid methyl esters. AB - Thermodynamic analysis of hydrophobic interaction chromatography of amino acid methyl esters showed entropy-driven adsorption, consistent with solvophobic theory, except for phenyl ester on the Toyopearl resins. All esters adsorbed more strongly to the Toyopearl resins, including the polymethacrylate base matrix, than to Butyl Sepharose. Enthalpy changes were more favorable with the former, explaining the retention difference between Toyopearl Butyl and Butyl Sepharose. An enthalpy change versus heat capacity change plot showed Van der Waals interactions predominantly with the resin matrix. Literature data revealed the same effect for dansylamino acids, shown by isothermodynamic temperature analysis to adsorb more entropically than the esters. PMID- 17919647 TI - Liquid-liquid extraction/headspace/gas chromatographic/mass spectrometric determination of benzene, toluene, ethylbenzene, (o-, m- and p-)xylene and styrene in olive oil using surfactant-coated carbon nanotubes as extractant. AB - BTEX-S compounds are widely distributed in the environment and can be present in different foodstuffs, including olive oil. Taking into account the risks of the exposure to these compounds, analytical methods for their determination in different matrices are mandatory. In this paper, the use of surfactant-coated multiwalled carbon nanotubes as additive in liquid-liquid extraction is applied for the determination of single-ring aromatic compounds in olive oil samples. After sample treatment, the aqueous extracts are subsequently analyzed by headspace/gas chromatography/mass spectrometry allowing the determination of BTEX S within ca. 15 min. Each stage of the proposed LLE/HS/GC/MS configuration involves a selectivity enhancement avoiding the interference of other compounds of the sample matrix. Limits of detection were in the range 0.25 ng mL(-1) (obtained for ethylbenzene) and 0.43 ng mL(-1) (for benzene). The repeatability of the proposed method expressed as RSD varied between 1.9% (styrene) and 3.3% (benzene) (n=11). PMID- 17919648 TI - Determination of fenthion and oxidation products in personal protection equipment by gas chromatography. AB - A method for the co-extraction and simultaneous chromatographic determination of fenthion and its five oxidation products (or metabolites) fenoxon, fenoxon sulfoxide, fenoxon-sulfone, fenthion-sulfoxide, and fenthion-sulfone in personal protection equipment (PPE) of pesticide applicators was developed and validated. Capillary gas chromatography-nitrogen-phosphorus detection was used for the analytical determination of all the aforementioned compounds over the concentration range of 0.1-0.5 microg/mL. All necessary validation criteria of the method were met. The method was found to be highly selective, accurate, and precise, gave satisfactory recovery (>70%) and RSD values (<20%) for fenthion and its metabolites in all tested specimens of personal protection equipment and in air samplers. The limit of quantification (LOQ) and linearity were determined for the most relevant PPE (i.e. inner coverall) for the parent compound and for the metabolites. The LOQ values ranged from 0.05 to 0.1 ppb while the linearity in the tested range of 0.1-0.5 ppm had r(2)>0.994 for all analytes. PMID- 17919650 TI - Stripping analysis of mercury(II) ionic solutions under magneto-hydrodynamic convection. AB - Inorganic mercury(II) ions are ubiquitous contaminants of world water systems and thus their determination and removal from the environment are important. The effects of magnetic field on the stripping analysis of mercury(II) ionic solutions have been experimentally investigated. During the stripping analysis, a potential difference is applied across the working and reference electrodes positioned in the working sample and a current density transmits through the electrolyte solution. When the electrochemical cell is exposed to a magnetic field, provided by a permanent magnet, the interaction between the current density and the magnetic field induces Lorentz forces, which, in turn, induce fluid motion. The induced magneto-hydrodynamic (MHD) convection enhances the ionic mass transport during the deposition and stripping steps, which leads to larger anodic current during the stripping step, thus obtaining higher detection sensitivity during the determination of the mercury(II) ions. The Hg2+ ionic solutions with concentrations ranging from 1 nM to 1 microM in the presence and absence of supporting electrolyte, 30 mM nitric acid (HNO 3) and 0.1 M potassium nitrate (KNO 3), under various magnetic flux densities (B=0,0.27,0.53, and 0.71 T) were measured with a linear sweep stripping voltammetry (LSSV) technique. The experimental results demonstrated that the stripping signals of the Hg2+ ions are enhanced, respectively, more than 10 and 30% in the absence and presence of the supporting electrolyte under a magnetic flux density B=0.71 T as compared to the cases in the absence of the magnetic field with all other identical conditions. PMID- 17919649 TI - Development of a fast capillary electrophoresis method for determination of creatinine in urine samples. AB - The aim of this work was to develop a fast method using capillary electrophoresis for the determination of creatinine in human urine samples. The pH and constituents of the background electrolyte were selected by inspection of effective mobility of creatinine and candidate urine interferents versus pH curves. The tendency of the analyte to undergo electromigration dispersion and the buffer capacity were evaluated by the Peakmaster software and considered in the optimization of the background electrolyte, composed by 10 mmol L(-1) tris(hydroxymethyl)aminomethane and 20 mmol L(-1) 2-hydroxyisobutyric acid (HIBA) at pH 3.93. Separation was conducted in a fused-silica capillary (32 cm total length and 8.5 cm effective length, 50 microm I.D.), with short-end injection configuration and direct UV detection at 215 nm. The migration time of creatinine was only 22s. A few figures of merit of the method are as follows: good linearity in the concentration interval of 5-70 mg L(-1) (R(2)>0.99), limit of detection of 0.5 mg L(-1), inter-day precision better than 2.7% (n=9) and recovery in the range 99.0-103.7% at three concentration levels (50, 100 and 150 mg L(-1)). Urine samples were prepared by deproteination with acetonitrile (1:3 sample:acetonitrile, v/v), centrifugation and dilution of a deproteinated aliquot with 12.5 mmol L(-1) HIBA (1:4, v/v). Creatinine concentrations between 489 and 1063 mg L(-1) were obtained in the urine of four healthy volunteers. PMID- 17919651 TI - A microgenetic study of insightful problem solving. AB - An eight-session microgenetic study of acquisition of an insightful problem solving strategy was conducted. A total of 35 second graders who did not use this insightful strategy initially were assigned to two groups that differed in the frequency of problems likely to facilitate discovery and generalization of the strategy. Children in the facilitative problems group discovered the insightful strategy earlier, used it more often subsequently, and transferred it more often to novel problems than did those in the nonfacilitative problems group. Children generally discovered the insightful strategy on the most facilitative items and extended it progressively to items on which its advantages were smaller but still substantial. The results indicate that experience outside the experimental situation, as well as experience inside the experimental situation, influences use of new strategies. PMID- 17919652 TI - Determinants of frequency-dependent contraction and relaxation of mammalian myocardium. AB - An increase in heart rate is the primary mechanism that up-regulates cardiac output during conditions such as exercise and stress. When the heart rate increases, cardiac output increases due to (1) an increased number of beats per time period, and (2) the fact that myocardium generates a higher level of force. In this review, we focus on the underlying mechanisms that are at the basis of frequency-dependent activation of the heart. In addition to increased force development, the kinetics of both cardiac activation and relaxation are faster. This is crucial, as in between successive beats there is less time, and cardiac output can only be maintained if the ventricle can fill adequately. We will discuss the cellular mechanisms that are involved in the regulation of rate dependent changes in kinetics, with a focus on changes that occur in regulation of the intracellular calcium transient, and the changes in the myofilament responsiveness that occur when the heart rate changes. PMID- 17919653 TI - Measurements of single DNA molecule packaging dynamics in bacteriophage lambda reveal high forces, high motor processivity, and capsid transformations. AB - Molecular motors drive genome packaging into preformed procapsids in many double stranded (ds)DNA viruses. Here, we present optical tweezers measurements of single DNA molecule packaging in bacteriophage lambda. DNA-gpA-gpNu1 complexes were assembled with recombinant gpA and gpNu1 proteins and tethered to microspheres, and procapsids were attached to separate microspheres. DNA binding and initiation of packaging were observed within a few seconds of bringing these microspheres into proximity in the presence of ATP. The motor was observed to generate greater than 50 picoNewtons (pN) of force, in the same range as observed with bacteriophage phi29, suggesting that high force generation is a common property of viral packaging motors. However, at low capsid filling the packaging rate averaged approximately 600 bp/s, which is 3.5-fold higher than phi29, and the motor processivity was also threefold higher, with less than one slip per genome length translocated. The packaging rate slowed significantly with increasing capsid filling, indicating a buildup of internal force reaching 14 pN at 86% packaging, in good agreement with the force driving DNA ejection measured in osmotic pressure experiments and calculated theoretically. Taken together, these experiments show that the internal force that builds during packaging is largely available to drive subsequent DNA ejection. In addition, we observed an 80 bp/s dip in the average packaging rate at 30% packaging, suggesting that procapsid expansion occurs at this point following the buildup of an average of 4 pN of internal force. In experiments with a DNA construct longer than the wild type genome, a sudden acceleration in packaging rate was observed above 90% packaging, and much greater than 100% of the genome length was translocated, suggesting that internal force can rupture the immature procapsid, which lacks an accessory protein (gpD). PMID- 17919654 TI - Altered dynamics of DNA bases adjacent to a mismatch: a cue for mismatch recognition by MutS. AB - The structural deviations as well as the alteration in the dynamics of DNA at mismatch sites are considered to have a crucial role in mismatch recognition followed by its repair utilizing mismatch repair family proteins. To compare the dynamics at a mismatch and a non-mismatch site, we incorporated 2-aminopurine, a fluorescent analogue of adenine next to a G.T mismatch, a C.C mismatch, or an unpaired T, and at several other non-mismatch positions. Rotational diffusion of 2-aminopurine at these locations, monitored by time-resolved fluorescence anisotropy, showed distinct differences in the dynamics. This alteration in the motional dynamics is largely confined to the normally matched base-pairs that are immediately adjacent to a mismatch/ unpaired base and could be used by MutS as a cue for mismatch-specific recognition. Interestingly, the enhanced dynamics associated with base-pairs adjacent to a mismatch are significantly restricted upon MutS binding, perhaps "resetting" the cues for downstream events that follow MutS binding. Recognition of such details of motional dynamics of DNA for the first time in the current study enabled us to propose a model that integrates the details of mismatch recognition by MutS as revealed by the high-resolution crystal structure with that of observed base dynamics, and unveils a minimal composite read-out involving the base mismatch and its adjacent normal base pairs. PMID- 17919655 TI - From genetic diversity to metabolic unity: studies on the biosynthesis of aurafurones and aurafuron-like structures in myxobacteria and streptomycetes. AB - The myxobacterial polyketide secondary metabolites aurafuron A and B were identified by genome mining in the myxobacterial strain Stigmatella aurantiaca DW4/3-1. The compounds contain an unusual furanone moiety and resemble metabolites isolated from soil-dwelling and marine actinobacteria, a fungus and mollusks. We describe here the cloning and functional analysis of the aurafuron biosynthetic gene cluster, including site-directed mutagenesis and feeding studies using labeled precursors. The polyketide core of the aurafurones is assembled by a modular polyketide synthase (PKS). As with many such systems described from myxobacteria, the aurafuron PKS exhibits a number of unusual features, including the apparent iterative use of a module, redundant modules and domains, a trans acting dehydratase and the absence of a terminal thioesterase domain. Four oxidoreductases are encoded within the gene locus, some of which likely participate in formation of the furanone moiety via a Baeyer-Villiger type oxidation. Indeed, inactivation of a gene encoding a cytochrome P(450) monooxygenase completely abolished production of both compounds. We also compare the complete gene locus to biosynthetic gene clusters from two Streptomyces sp., which produce close structural analogues of the aurafurones. A portion of the post-PKS biosynthetic machinery is strikingly similar in all three cases, in contrast to the PKS genes, which are highly divergent. Phylogenetic analysis of the ketosynthase domains further indicates that the PKSs have developed independently (polyphyletically) during evolution. These findings point to a currently unknown but important biological function of aurafuron-like compounds for the producing organisms. PMID- 17919656 TI - Comparative normal mode analysis of LFA-1 integrin I-domains. AB - The conformational dynamics of the Inserted domain (I-domain) from the lymphocyte function-associated antigen-1 (LFA-1) was investigated by normal mode analysis of multiple structures of the low, intermediate, and high affinity states. LFA-1 is an integrin expressed on leukocytes and is of critical importance in adhesion reactions, like antigen-specific responses, homing, and diapedesis. The main ligand binding site of LFA-1 is the I-domain, which recognizes intercellular adhesion molecules (ICAMs), members of the immunoglobulin superfamily. From experimental crystal structures, a large-scale conformational change of, among others, the alpha7 helix of the I-domain has been observed leading to the proposal that these structural changes are linked to the conformational regulation of LFA-1. The results from the present calculations show that structural changes of the alpha7 helix consistent with those observed in the crystal structures are significantly sampled by the low frequency modes. This was found to be particularly true for the low affinity state of the I-domain, indicating that low frequency motions favor the conformational transition implicated in activation. However, beyond the simple downward shift of the helix implied by the crystal structures, the calculations further show that there is a noticeable swinging-out motion of the helix. The consequences of this motion are discussed in the context of integrin activation and inhibition. Moreover, significant changes in the atomic-level dynamics and in long-range correlated motions of the I-domain were found to occur upon binding of the natural ligand ICAM. These changes were more local upon binding of an allosteric inhibitor. The present study opens the question of how changes in dynamics may contribute to the long-range transmission of signal upon ICAM binding by the LFA-1 I-domain. PMID- 17919657 TI - Gal11p dosage-compensates transcriptional activator deletions via Taf14p. AB - Transcriptional activators work by recruiting transcription factors that are required for the process of transcription to their target genes. We have used the Split-Ubiquitin system to identify eight transcription factors that interacted with both the transcriptional activators Gal4p and Gcn4p in living cells. The over-expression of one of the activator-interacting proteins, Gal11p, partially suppressed GAL4 and GCN4 deletions. We have isolated two point mutants in Gal11p, F848L and F869S that were defective for the dosage compensation. We have identified 35 transcription factors that interacted with Gal11p in living cells, and the only protein-protein interaction affected by the Gal11p mutations was the one between Gal11p and Taf14p. We have further shown that the suppression of a GAL4 deletion by high levels of Gal11p required Taf14p, and that over-expression of Gal11p recruited Taf14p to the GAL1 promoter together with Tbp1p, Swi2p and Srb7p. Gal11p interacted with Mig1p, indicating that Mig1/2p could have recruited Gal11p to the GAL1 promoter in the absence of Gal4p. Our results suggest that transcriptional activators work by raising the local concentration of the limiting factor Gal11p, and that Gal11p works by recruiting Mediator and Taf14p containing transcription factors like TFIID and SWI/SNF and by competing general repressors like Ssn6p-Tup1p off the target promoters. PMID- 17919659 TI - Fatty acids can substitute the HIV fusion peptide in lipid merging and fusion: an analogy between viral and palmitoylated eukaryotic fusion proteins. AB - Various fusion proteins from eukaryotes and viruses share structural similarities such as a coiled coil motif. However, compared with eukaryotic proteins, a viral fusion protein contains a fusion peptide (FP), which is an N-terminal hydrophobic fragment that is primarily involved in directing fusion via anchoring the protein to the target cell membrane. In various eukaryotic fusion proteins the membrane targeting domain is cysteine-rich and must undergo palmitoylation prior to the fusion process. Here we examined whether fatty acids can replace the FP of human immunodeficiency virus type 1 (HIV-1), thereby discerning between the contributions of the sequence versus hydrophobicity of the FP in the lipid merging process. For that purpose, we structurally and functionally characterized peptides derived from the N terminus of HIV fusion protein - gp41 in which the FP is lacking or replaced by fatty acids. We found that fatty acid conjugation dramatically enhanced the capability of the peptides to induce lipid mixing and aggregation of zwitterionic phospholipids composing the outer leaflet of eukaryotic cell membranes. The enhanced effect of the acylated peptides on membranes was further supported by real-time atomic force microscopy (AFM) showing nanoscale holes in zwitterionic membranes. Membrane-binding experiments revealed that fatty acid conjugation did not increase the affinity of the peptides to the membrane significantly. Furthermore, all free and acylated peptides exhibited similar alpha-helical structures in solution and in zwitterionic membranes. Interestingly, the fusogenic active conformation of N36 in negatively charged membranes composing the inner leaflet of eukaryotic cells is beta-sheet. Apparently, N-terminal heptad repeat (NHR) can change its conformation as a response to a change in the charge of the membrane head group. Overall, the data suggest an analogy between the eukaryotic cysteine-rich domains and the viral fusion peptide, and mark the hydrophobic nature of FP as an important characteristic for its role in lipid merging. PMID- 17919658 TI - Critical role of mac-1 sialyl lewis x moieties in regulating neutrophil degranulation and transmigration. AB - Leukocyte cell surface sialyl Lewis x (sLe(x)) and related epitopes play an important role in cell rolling and adhesion during diapedesis via interaction with E-selectin. Here, we present evidence that Mac-1 (CD11b/CD18, CR-3) is a major neutrophil glycoprotein decorated with sLe(x) and ligation of these carbohydrate moieties by anti-sLe(x) antibody significantly impairs neutrophil functions. First, Western blot analysis shows that both CD11b and CD18 subunit of purified Mac-1 are decorated with sLe(x) moieties. A significant co-localization of CD11b and sLe(x) moieties is observed at neutrophil secondary granules. With stimulation of formyl-Met-Leu-Phe (fMLP), neutrophil surface labeling with anti sLe(x) antibody follows an identical up-regulation pattern of Mac-1. Second, protein-binding assays indicate that sLe(x) moieties on Mac-1 are critical for binding interaction of Mac-1 to E-selectin. Removal of sLe(x) moieties completely abolishes Mac-1-E-selectin binding. Finally, ligation of Mac-1 sLe(x) by anti sLe(x) antibody induces a significant degranulation of neutrophil secondary granules at the absence of chemoattractant stimulation. This "dysregulated" degranulation induced by anti-sLe(x) antibody strongly inhibits neutrophil transmigration in response to fMLP. In summary, Mac-1 sLe(x) moieties play a critical role in regulating beta(2) integrin functions during neutrophil transmigration and degranulation. PMID- 17919660 TI - Model for RuvAB-mediated branch migration of Holliday junctions. AB - During RuvAB-mediated Holliday-junction migration two opposite arms of double stranded DNA (dsDNA) are driven to translocate unidirectional by two respective ring-like hexameric RuvB proteins. However, how the RuvB protein, powered by ATP hydrolysis, drives unidirectional translocation of dsDNA is not clear. Here a model is presented for this mechanochemical-coupling mechanism. In the model, the unidirectional translocation is resulted from both the ATP hydrolysis-induced rotation (power stroke) of the RuvB subunits and the passage of the strong DNA binding from the previous to next RuvB subunits during the sequential ATPase activities around the ring. Using the model, the relationship between the power stroke size, the step size of DNA translocation and the ratio of the rotational rate of DNA over that of RuvB relative to RuvA is predicted. PMID- 17919661 TI - Sperm competition can drive a male-biased mutation rate. AB - A pattern of male-biased mutation has been found in a wide range of species. The standard explanation for this bias is that there are greater numbers of mitotic cell divisions in the history of the average sperm, compared to the average egg, and that mutations typically result from errors made during replication. However, this fails to provide an ultimate evolutionary explanation for why the male germline would tolerate more mutations that are typically deleterious. One possibility is that if there is a tradeoff between producing large numbers of sperm and expending energetic resources in maintaining a lower mutation rate, sperm competition would select for males that produce larger numbers of sperm despite a higher resulting mutation rate. Here I describe a model that jointly considers the fitness consequences of deleterious mutation and mating success in the face of sperm competition. I show that a moderate level of sperm competition can account for the observation that the male germline tolerates a higher mutation rate than the female germline. PMID- 17919662 TI - Optimizing bacteriophage plaque fecundity. AB - Bacteriophages (phages), the viruses of bacteria, form visible lesions within bacterial lawns (called plaques), which are employed ubiquitously in phage isolation and characterization. Plaques also can serve as models for phage population growth within environments that display significant spatial structure, e.g. soils, sediments, animal mucosal tissue, etc. Furthermore, phages growing within plaques, in experimental evolution studies, may become adapted to novel conditions, may be selected for faster expansion, or may evolve toward producing more virions per plaque. Here, we examine the evolution of the latter, greater plaque fecundity, considering especially tradeoffs between phage latent period and phage burst size. This evolution is interesting because genetically lengthening latent periods, as seen with phage lysis-timing mutants, should increase phage burst sizes, as more time is available for phage-progeny maturation during infection. Genetically shortening latent periods, however, is a means toward producing larger phage plaques since phage virions then can spend more time diffusing rather than infecting. With these larger plaques more bacteria become phage infected, resulting in more phage bursts. Given this conflict between latent period's impact on per-plaque burst number versus per infection burst size, and based on analysis of existing models of plaque expansion, we provide two assertions. First, latent periods that optimize plaque fecundity are longer (e.g. at least two-fold longer) than latent periods that optimize plaque size (or that optimize phage population growth within broth). Second, if increases in burst size can contribute to plaque size (i.e. larger plaques with larger bursts), then latent-period optima that maximize plaque fecundity should be longer still. As a part of our analysis, we provide a means for predicting latent-period optima-for maximizing either plaque size or plaque fecundity-which is based on knowledge of only phage eclipse period and the relative contribution of phage burst size versus latent period toward plaque size. PMID- 17919663 TI - Pigment epithelium-derived factor inhibits glioma cell growth in vitro and in vivo. AB - Glioblastoma multiforme is the most common malignant brain tumor in adults, and it is among the most lethal of all cancers. Recent studies have shown that pigment epithelium-derived factor (PEDF) can induce differentiation and inhibit angiogenesis of several tumors. This study was designed to determine whether gliomas angiogenesis and tumor growth could be inhibited by PEDF. We found that PEDF down-regulated expression levels of vascular endothelial growth factor and up-regulated the expression of thrombospondin-2 and augmented apoptosis in a dose dependent manner in both A172 and U87 glioma cells lines after 48 h of treatment. Analysis of the cell cycle showed arrest in the G(1) phase and block in S phase of the cell cycle. Meanwhile PEDF induced apoptosis was associated with increases of p53 and Bax and inhibition of Bcl-2. Conditioned medium with PEDF showed a significantly reductive effect on migration in vitro accompanied with a significant reduction of matrix metalloproteinase-9 expression. PEDF suppressed glioma cell migration in vitro and tumor burden in athymic nude mice. These results demonstrate for the first time inhibitory effects of PEDF on the growth and migration of human gliomas via induction of apoptosis and blocking of migratory-related factors. PEDF activation can be a novel approach for future therapeutic purposes against gliomas. PMID- 17919664 TI - Mutagenic radioadaptation in a human lymphoblastoid cell line. AB - We investigated the mutagenic radioadaptive response of human lymphoblastoid TK6 cells by pretreating them with a low dose (5 cGy) of X-rays followed by a high (2 Gy) dose 6h later. Pretreatment reduced the 2-Gy-induced mutation frequency (MF) of the thymidine kinase (TK) gene (18.3 x 10(-6)) to 62% of the original level (11.4 x 10(-6)). A loss of heterozygosity (LOH) detection analysis applied to the isolated TK(-) mutants revealed the mutational events as non-LOH (resulting mostly from a point mutation in the TK gene), hemizygous LOH (resulting from a chromosomal deletion), or homozygous LOH (resulting from homologous recombination (HR) between chromosomes). For non-LOH events, pretreatment decreased the frequency to 27% of the original level (from 7.1 x 10(-6) to 1.9 x 10(-6)). cDNAs prepared from the non-LOH mutants revealed that the decrease was due mainly to the repression of base substitutions. The frequency of hemizygous LOH events, however, was not significantly altered by pretreatment. Mapping analysis of chromosome 17 demonstrated that the distribution and the extent of hemizygous LOH events were also not significantly influenced by pretreatment. For homozygous LOH events, pretreatment reduced the frequency to 61% of the original level (from 5.1 x 10(-6) to 3.1 x 10(-6)), reflecting an enhancement in HR repair of DNA double strand breaks. Our findings suggest that the radioadaptive response in TK6 cells follows mainly from mutations at the base-sequence level, not the chromosome level. PMID- 17919665 TI - Involvement of brain endogenous histamine in the degeneration of dopaminergic neurons in 6-hydroxydopamine-lesioned rats. AB - Previous studies have suggested that brain histamine is involved in the pathogenesis of Parkinson's disease (PD), but the role of endogenous histamine in the degeneration of dopaminergic neurons in the substantia nigra pars compact (SNpc) remains unclear. We aimed to investigate this issue by changing the brain histamine levels by giving histaminergic agents, and administrating histamine receptor antagonists in the PD animal model, i.e. the 6-hydroxydopamine (6-OHDA) lesioned rat. In saline-treated animals, 6-OHDA infusion produced a progressive increase in apomorphine-induced turning rate and a loss of tyrosine hydroxylase immunoreactive (TH-ir) neurons in the SNpc. Histaminergic agents were given prior and daily for 1, 7 or 14 days after 6-OHDA infusion. Histidine (500 mg/kg, i.p.), a precursor of histamine, increased the turning rate (27% on day 7 and 26% on day 14, respectively; P<0.05) and also the loss of TH-ir neurons, but only on day 1 and 7 (67% vs 47% and 90.4% vs 74% loss, respectively; P<0.05). In contrast, alpha-fluoromethylhistidine (alpha-FMH, 25 microg, i.c.v.), an irreversible inhibitor of histidine decarboxylase (HDC), significantly decreased the turning rate (25% on day 7 and 26% on day 14, respectively; P<0.05) and prevented the loss of TH-ir neurons, also only on day 1 and day 7 (28% vs 47% and 58% vs 74% loss, respectively; P<0.05). In addition, the histamine H(1) receptor antagonist pyrilamine (5 microg, i.c.v.), but not the H(2) receptor antagonist cimetidine (5 microg, i.c.v.), also decreased the turning rate (38% on day 7 and 21% on day 14, respectively; P<0.05) and prevented the loss of TH-ir neurons on day 1 and day 7 (38% vs 51% and 60% vs 78% loss, respectively; P<0.05). On day 14 after 6-OHDA lesion, there were no significant differences in the number of TH-ir neurons among all the different treatment groups. Taken together, these findings indicate that endogenous histamine may accelerate the degeneration of dopaminergic neurons via its H(1) receptor, while attenuation of histamine transmission may play a protective role on it in the early stage of development of 6-OHDA lesioned PD rats. PMID- 17919666 TI - Plant bioactives for ruminant health and productivity. AB - Plants have been used throughout history for their medicinal properties. This use has often focused on human health but plants have also been, and still are, applied in ethnoveterinary practice and animal health management. In recent times, the use of synthetic chemicals has become prevalent. Public awareness of the potential environmental and health risks associated with heavy chemical use has also increased. This has put pressure on regulatory bodies to reduce the use of chemicals in agriculture. The most striking example is the 2006 banning of antibiotics in animal feed by the European Union. Moves such as this have increased the drive to find alternatives to synthetic chemicals and research has again turned to the use of plant bioactives as a means of improving animal health. Current scientific evidence suggests there is significant potential to use plants to enhance animal health in general and that of ruminants (cattle, deer, sheep, etc.) in particular. Active areas of research for plant bioactives (particularly saponin and tannin containing plants) include reproductive efficiency, milk and meat quality improvement, foam production/bloat control and methane production. Nematode control is also a significant area of research and the evidence suggests a much broader range of phytochemicals may be effective. This review presents a summary of the literature and examines international research efforts towards the development of plant bioactives for animal health. PMID- 17919668 TI - Effects of water deprivation for 72 h on the pharmacokinetics of ipriflavone in rats. AB - It has been reported that ipriflavone was primarily metabolized via hepatic microsomal cytochrome P450 (CYP) 1A1/2 and 2C11 in rats, and the expression of hepatic CYP1A2 and 2C11 was not changed in rats with water deprivation for 72h compared to controls. Hence, it could be expected that the time-averaged nonrenal clearance (Cl(nr)) of ipriflavone would be comparable between two groups of rat. As expected, after intravenous administration of ipriflavone (20 mg/kg), the Cl(nr) values of ipriflavone were comparable between two groups of rat. This could be supported by comparable in vitro intrinsic clearance (Cl(int)) values for the disappearance of ipriflavone in hepatic microsomes for both groups of rat. After oral administration of ipriflavone (200 mg/kg), the total area under the plasma concentration-time curve from time zero to the last measured time, 12h, in plasma (AUC(0-12h)) values of ipriflavone were also comparable between two groups of rat. The above data suggest that dehydration state did not affect significantly pharmacokinetics of ipriflavone in rats. PMID- 17919669 TI - [Alveolar rhabdomyosarcoma of the parotid gland]. AB - INTRODUCTION: Alveolar rhabdomyosarcoma of salivary glands is a rare malignant neoplasm, presenting a striated muscular differentiation. It is typically observed in children, adolescents, and young adults but can sometimes be diagnosed in older people. OBSERVATION: We report the case of a 57 year-old man presenting with a primary alveolar rhabdomyosarcoma of the parotid gland. The patient was treated by surgery and radiotherapy. DISCUSSION: According to literature data, alveolar rhabdomyosarcoma of the parotid gland is exceptionally described after 40 years of age. In that case it has the worst prognosis and its lethality is due mainly to its very early metastasis. PMID- 17919670 TI - [Piezoelectric osteotomy: applications in periodontal and implant surgery]. AB - Reverse piezoelectricity is currently used in Dentistry for ultrasonic scaling devices. Using this technology, increasingly more powerful ultrasonic surgical lancets were developed within a few years, and these new tools have provided many practical solutions in oral and maxillofacial surgery. This article reviews the basic principles of piezosurgery, and its numerous clinical applications: in oral surgery (atraumatic extractions, wisdom teeth exposure, periapical cyst debridement, pre-prosthetic surgery), more specifically in periodontal (root planning and bone remodeling, crown lengthening) and implant surgery (sinus lift, removal of fractured implants, bone ridge augmentation, bone graft harvesting (crestal, retromolar, chin or tori)). The recent increased power of these instruments allows emphasizing a much wider range of clinical applications, extending to all surgical fields. PMID- 17919671 TI - Hookworm infections in Singaporean soldiers after jungle training in Brunei Darussalam. AB - In June 2006, 118 Singaporean soldiers visited Brunei for jungle training for 10 days. Subsequently, two soldiers had severe diarrhoea and were diagnosed with severe hookworm infection. An epidemiological investigation and case-control study was conducted among the 118 soldiers. A survey was administered to determine the exposures predisposing to infection. Laboratory tests included a full blood count and three separate faecal screens via microscopy and culture. All servicemen were treated with one dose of mebendazole, and follow-up screening was performed 3 months later. Of 113 soldiers completing the questionnaire, 43 soldiers (38%) reported gastrointestinal symptoms post deployment, with 33 (29%) complaining of diarrhoea and 29 (26%) of abdominal pain. Eight had an itch or rash, but none were suggestive of cutaneous larva migrans. Of 103 soldiers completing both the questionnaire and with all the laboratory tests, 42 soldiers (41%) had eosinophilia (>0.6x10(9)/l) and 18 (17%) had hookworm infection on microscopy. More than 89% recalled substantial exposure to soil or groundwater, but no exposure was significantly associated with eosinophilia or infection. After adjusting for possible exposures, not wearing footwear during rest periods had a significantly higher odds ratio (2.86) for acquiring hookworm infection or eosinophilia. Helminth infections are common in visitors to rural Asia. As preventing exposure is often difficult, empirical treatment with anthelminthics should be considered in travellers to high risk areas. PMID- 17919672 TI - Malaria diagnosis under field conditions in the Venezuelan Amazon. AB - To improve practical, accurate diagnosis of malaria in the Amazon rainforest of Venezuela, two rapid diagnostic tests (RDT) (OptiMAL-IT) and FalciVax) and a laboratory light microscope, used in the field with a battery-operated head lamp as an external light source, were evaluated against the standard laboratory microscope procedure for malaria detection. One hundred and thirty-six Yanomami patients were studied for the presence of malaria parasites. Thirty-three patients (24%) were positive for malaria (Plasmodium falciparum, P. vivax, P. malariae). Twenty-one (64%) of the positive patients had <100 parasites/microl. Both RDTs showed poor sensitivity (24.2% for OptiMAL-IT) and 36.4% for FalciVax) but good specificity (99% both for OptiMAL-IT) and FalciVax). Field and laboratory microscopy showed sensitivities of 94% and 91%, respectively. The kappa coefficient was 0.90, indicating a high agreement between field and laboratory microscopy. We conclude that (i) adequate slide reading cannot be substituted by either of the two RDTs in the Venezuelan Amazon and (ii) the use of a light source such as that described above makes slide reading more feasible than hitherto in remote areas without electricity. PMID- 17919673 TI - Subhepatotoxic exposure to arsenic enhances lipopolysaccharide-induced liver injury in mice. AB - Exposure to arsenic via drinking water is a serious health concern in the US. Whereas studies have identified arsenic alone as an independent risk factor for liver disease, concentrations of arsenic required to damage this organ are generally higher than found in the US water supply. The purpose of the current study was to test the hypothesis that arsenic (at subhepatotoxic doses) may also sensitize the liver to a second hepatotoxin. To test this hypothesis, the effect of chronic exposure to arsenic on liver damage caused by acute lipopolysaccharide (LPS) was determined in mice. Male C57Bl/6J mice (4-6 weeks) were exposed to arsenic (49 ppm as sodium arsenite in drinking water). After 7 months of exposure, animals were injected with LPS (10 mg/kg i.p.) and sacrificed 24 h later. Arsenic alone caused no overt hepatotoxicity, as determined by plasma enzymes and histology. In contrast, arsenic exposure dramatically enhanced liver damage caused by LPS, increasing the number and size of necroinflammatory foci. This effect of arsenic was coupled with increases in indices of oxidative stress (4-HNE adducts, depletion of GSH and methionine pools). The number of apoptotic (TUNEL) hepatocytes was similar in the LPS and arsenic/LPS groups. In contrast, arsenic pre-exposure blunted the increase in proliferating (PCNA) hepatocytes caused by LPS; this change in the balance between cell death and proliferation was coupled with a robust loss of liver weight in the arsenic/LPS compared to the LPS alone group. The impairment of proliferation after LPS caused by arsenic was also coupled with alterations in the expression of key mediators of cell cycle progression (p27, p21, CDK6 and Cyclin D1). Taken together, these results suggest that arsenic, at doses that are not overtly hepatotoxic per se, significantly enhances LPS-induced liver injury. These results further suggest that arsenic levels in the drinking water may be a risk modifier for the development of chronic liver diseases. PMID- 17919674 TI - Overexpression of cerebral and hepatic cytochrome P450s alters behavioral activity of rat offspring following prenatal exposure to lindane. AB - Oral administration of different doses (0.0625, 0.125 or 0.25 mg/kg corresponding to 1/1400th, 1/700th or 1/350th of LD(50)) of lindane to the pregnant Wistar rats from gestation days 5 to 21 were found to produce a dose-dependent increase in the activity of cytochrome P450 (CYP)-dependent 7-ethoxyresorufin-O-deethylase (EROD), 7-pentoxyresorufin-O-dealkylase (PROD) and N-nitrosodimethylamine demethylase (NDMA-d) in brain and liver of offspring postnatally at 3 weeks. The increase in the activity of CYP monooxygenases was found to be associated with the increase in the mRNA and protein expression of xenobiotic metabolizing CYP1A, 2B and 2E1 isoenzymes in the brain and liver of offspring. Dose-dependent alterations in the parameters of spontaneous locomotor activity in the offspring postnatally at 3 weeks have suggested that increase in CYP activity may possibly lead to the formation of metabolites to the levels that may be sufficient to alter the behavioral activity of the offspring. Interestingly, the inductive effect on cerebral and hepatic CYPs was found to persist postnatally up to 6 weeks in the offspring at the relatively higher doses (0.125 and 0.25 mg/kg) of lindane and up to 9 weeks at the highest dose (0.25 mg/kg), though the magnitude of induction was less than that observed at 3 weeks. Alterations in the parameters of spontaneous locomotor activity in the offspring postnatally at 6 and 9 weeks, though significant only in the offspring at 3 and 6-week of age, have further indicated that due to the reduced activity of the CYPs during the ontogeny, lindane and its metabolites may not be effectively cleared from the brain. The data suggest that low dose prenatal exposure to the pesticide has the potential to produce overexpression of xenobiotic metabolizing CYPs in brain and liver of the offspring which may account for the behavioral changes observed in the offspring. PMID- 17919676 TI - Pharmacological cdk inhibitor R-Roscovitine suppresses JC virus proliferation. AB - The human Polyomavirus JC virus (JCV) utilizes cellular proteins for viral replication and transcription in the host cell nucleus. These cellular proteins represent potential targets for antiviral drugs against the JCV. In this study, we examined the antiviral effects of the pharmacological cyclin-dependent kinase (cdk) inhibitor R-Roscovitine, which has been shown to have antiviral activity against other viruses. We found that Roscovitine significantly inhibited the viral production and cytopathic effects of the JCV in a JCV-infected cell line. Roscovitine attenuated the transcriptional activity of JCV late genes, but not early genes, and also prevented viral replication via inhibiting phosphorylation of the viral early protein, large T antigen. These data suggest that the JCV requires cdks to transcribe late genes and to replicate its own DNA. That Roscovitine exhibited antiviral activity in JCV-infected cells suggests that Roscovitine might have therapeutic utility in the treatment of progressive multifocal leukoencephalopathy (PML). PMID- 17919675 TI - Induction of CYP1A1 and CYP1B1 by benzo(k)fluoranthene and benzo(a)pyrene in T 47D human breast cancer cells: roles of PAH interactions and PAH metabolites. AB - The interactions of polycyclic aromatic hydrocarbons (PAH) and cytochromes P450 (CYP) are complex; PAHs are enzyme inducers, substrates, and inhibitors. In T-47D breast cancer cells, exposure to 0.1 to 1 microM benzo(k)fluoranthene (BKF) induced CYP1A1/1B1-catalyzed 17beta-estradiol (E(2)) metabolism, whereas BKF levels greater than 1 muM inhibited E(2) metabolism. Time course studies showed that induction of CYP1-catalyzed E(2) metabolism persisted after the disappearance of BKF or co-exposed benzo(a)pyrene, suggesting that BKF metabolites retaining Ah receptor agonist activity were responsible for prolonged CYP1 induction. BKF metabolites were shown, through the use of ethoxyresorufin O deethylase and CYP1A1-promoter-luciferase reporter assays to induce CYP1A1/1B1 in T-47D cells. Metabolites formed by oxidation at the C-2/C-3 region of BKF had potencies for CYP1 induction exceeding those of BKF, whereas C-8/C-9 oxidative metabolites were somewhat less potent than BKF. The activities of expressed human CYP1A1 and 1B1 with BKF as substrate were investigated by use of HPLC with fluorescence detection, and by GC/MS. The results showed that both enzymes efficiently catalyzed the formation of 3-, 8-, and 9-OHBKF from BKF. These studies indicate that the inductive effects of PAH metabolites as potent CYP1 inducers are likely to be additional important factors in PAH-CYP interactions that affect metabolism and bioactivation of other PAHs, ultimately modulating PAH toxicity and carcinogenicity. PMID- 17919677 TI - Baffled membrane bioreactor (BMBR) for efficient nutrient removal from municipal wastewater. AB - Submerged membrane bioreactors (MBRs) are now widely used for various types of wastewater treatment. One drawback of submerged MBRs is the difficulty in removing nitrogen because intensive aeration is usually carried out in the tank and the MBRs must therefore be operated under aerobic conditions. In this study, the feasibility of treating municipal wastewater by a baffled membrane bioreactor (BMBR), particularly in terms of nitrogen removal, was examined. Simultaneous nitrification/denitrification in a single and small reaction tank was possible by inserting baffles into a normal submerged MBR as long as wastewater was fed in the appropriate way. To examine the applicability of the BMBR, pilot-scale experiments were carried out using real municipal wastewater. Although neither external carbon addition nor mixed liquor circulation was carried out in the operation of the BMBR, average removal rates of total organic carbon (TOC), total phosphorus (T-P) and total nitrogen (T-N) reached 85%, 97% and 77%, respectively, with the hydraulic retention time (HRT) of 4.7h. Permeability of the membrane could be maintained at a high level throughout the operation. It was found that denitrification was the limiting step in removal of nitrogen in the BMBR in this study. Various types of monitoring carried out in the BMBR also demonstrated the possibility of further improvements in its performance. PMID- 17919678 TI - Role of Fe(II), phosphate, silicate, sulfate, and carbonate in arsenic uptake by coprecipitation in synthetic and natural groundwater. AB - Competitive effects of phosphate, silicate, sulfate, and carbonate on As(III) and As(V) removal at pH approximately 7.2 have been investigated to test the feasibility of Fe(II)(aq) and hydroxylapatite crystals as inexpensive and potentially efficient agents for remediation of contaminated well-water, using Bangladesh as a type study. Arsenic(III) removal approximately 50-55% is achieved, when Fe(II)(aq) oxidizes to Fe(III) and precipitates as Fe(OH)3 at 25 degrees C and 3h reaction time, in the presence of all the oxyanion. Similar results were obtained for well-water samples from two sites in Bangladesh. Heating at 95 degrees C for 24h results in 70% As(III) uptake due to precipitation of magnesian calcite. A two-step process, Fe(II) oxidation and Fe(OH)3 precipitation at 25 degrees C for 2h, followed by magnesian calcite precipitation at 95 degrees C for 3h, yields approximately 65% arsenic removal while reducing the expensive heating period. In the absence of silicate, up to 70% As(III) uptake occurs at 25 degrees C. In all cases, As(III) was oxidized to As(V) in solution by dissolved oxygen and the reaction rate was probably promoted by intermediates formed during Fe(II) oxidation. Iron-catalyzed oxidation of As(III) by oxygen and hydrogen peroxide is pH-dependent with formation of oxidants in the Fenton reaction. Buffering pH at near-neutral values by dissolved carbonate and hydroxylapatite seeds is important for faster Fe(II) oxidation kinetics ensuring rapid coprecipitation of As as As(V) in the ferric phases. PMID- 17919679 TI - Removal of carbon disulfide via coupled reactions on a bi-functional catalyst: experimental and modeling results. AB - A mathematical model describing the rate of carbon disulfide (CS(2)) removal due to coupled reactions has been developed. Kinetic studies were carried out in a fixed bed reactor under atmospheric pressure and a range of temperatures (85-125 degrees C). The effects of flow rate, CS(2) inlet concentration, temperature and relative humidity were analyzed. A kinetic model based on axial dispersion, external and internal mass-transfer resistances, as well as effects of S deposition on the inner-face of the catalyst was in agreement with the CS(2) experimental breakthrough curves. The mathematical model can be used for process design and scale up of similar systems. PMID- 17919680 TI - Estimating half-lives of pesticides in/on vegetation for use in multimedia fate and exposure models. AB - Degradation half-lives in/on vegetation are needed in environmental risk assessment of pesticides, but these data are often not available for most active ingredients. To address this, we first correlated experimental soil degradation half-life data of 41 pesticides obtained from the reviewed literature with the corresponding experimental half-lives on plant surface. Degradation half-lives in soil were found to be four times slower compared with half-lives on plant surfaces. In a second step, we explored measured plant surface half-lives directly with those in vegetation. The results were validated by comparing computed values with results obtained from an experimental set-up. The uptake and dissipation of alpha-cypermethrin (insecticide) and bromopropylate (acaricide) was studied by detecting pesticide residues in whole and peeled tomato fruits using gas chromatography. Half-lives within vegetation were found to be four times faster compared with plant surface half-lives. Using this experimental based approach, it is concluded that the estimation of degradation half-lives of pesticides in/on vegetation to be used as input data in environmental mass balance models can be directly correlated from the more abundant ready experimental degradation half-life data for soil. PMID- 17919681 TI - Restoration of cadmium-contaminated paddy soils by washing with ferric chloride: Cd extraction mechanism and bench-scale verification. AB - The ability of FeCl3 to extract Cd from three paddy soils was compared with that of various irons, manganese, and zinc salts to elucidate the extraction mechanism. Manganese, zinc and iron salts (including FeCl3) extracted 4-41%, 8 44% and 24-66% of total Cd, respectively. This difference reflected the pH of the extraction solution, indicating that the primary mechanism of Cd extraction by FeCl3 is proton release coupled with hydroxide generation, as iron hydroxides are insoluble. Washing with FeCl3 led to the formation of Cd-chloride complexes, enhancing Cd extraction from the soils. FeCl3 effectively extracted Cd from all of the three soils compared to HCl that is a conventional washing chemical, when the concentrations of the two washing chemicals were between 15 and 60mM(c) (at above extraction pH 2.4), while the corresponding extraction pH of FeCl3 was slightly higher than HCl. As HCl is the strong acid of complete dissociation, if excess amount of HCl was added to soil, it is possible to give the dissolution of clay minerals in soils. In contrast, proton release from FeCl3 is controlled by the chemical equilibrium of hydroxide formation. While soil fertility properties were affected by a bench-scale soil washing with 45mM(c) FeCl3, adverse effects were not crucial and could be corrected. The bench-scale test confirmed the effectiveness of FeCl3 for removal of soil Cd. The washing had no negative effect on rice yield and lowered the Cd concentration of rice grain and rice straw in a pot experiment. PMID- 17919682 TI - Clear effects of soil organic matter chemistry, as determined by NMR spectroscopy, on the sorption of diuron. AB - Organic matter has long been recognized as the main sorbent phase in soils for hydrophobic organic compounds (HOCs). In recent times, there has been an increasing realization that not only the amount, but also the chemical composition, of organic matter can influence the sorption properties of a soil. Here, we show that the organic carbon-normalized sorption coefficient (K(OC)) for diuron is 27-81% higher in 10 A11 horizons than in 10 matching A12 horizons for soils collected from a small (2ha) field. K(OC) was generally greater for the deeper (B) horizons, although these values may be inflated by sorption of diuron to clays. Organic matter chemistry of the A11 and A12 horizons was determined using solid-state 13C nuclear magnetic resonance (NMR) spectroscopy. K(OC) was positively correlated with aryl C (r2=0.59, significance level 0.001) and negatively correlated with O-alkyl C (r2=0.84, significance level <0.001). This is only the second report of correlations between whole soil K(OC) and NMR derived measures of organic matter chemistry. We suggest that this success may be a consequence of limiting this study to a very small area (a single field). There is growing evidence that interactions between organic matter and clay minerals strongly affect K(OC). However, because the soil mineralogy varies little across the field, the influence of these interactions is greatly diminished, allowing the effect of organic matter chemistry on K(OC) to be seen clearly. This study in some way reconciles studies that show strong correlations between K(OC) and the chemistry of purified organic materials and the general lack of such correlations for whole soils. PMID- 17919683 TI - Influences of preparative methods of humic acids on the sorption of 2,4,6 trichlorophenol. AB - Humic acids (HAs) are a major component of soil organic matter which strongly affects the sorption behavior of organic contaminants in soils. To assess the sorption-desorption characteristics of organic compounds on HAs, the organic adsorbent is usually isolated using an acid-base extraction method followed by air-drying or freeze-drying. In this study, a peat soil from the Yangming mountain area of Taiwan was sampled and repeatedly extracted followed by either air-drying or a non-drying treatment (denoted DHAs and NDHAs, respectively). The sorption of 2,4,6-TCP on HAs was evaluated using the batch method. Kinetic sorption results indicated that DHAs exhibited a two-step first-order sorption behavior, involving a rapid sorption followed by a slow sorption. The slow sorption may be attributed to the diffusion of 2,4,6-TCP through the condensed aromatic domains of HAs. On the contrary, the sorption of 2,4,6-TCP on NDHAs was extremely rapid, and the sorption data did not fit existing kinetic models. Each HA sample exhibited a nonlinear sorption isotherm. Sorption nonlinearity (represented by Freundlich N values) and K(oc) had a positive relationship with aliphaticity for DHAs; however, nonlinearity and K(oc) correlated positively with aromaticity when NDHAs adsorbents were used. We conclude that the air-drying technique may artificially create a more condensed area, which strongly affects the sorption characteristics of HAs. Thus, an incorrect evaluation of the sorption capacity and its relationship with the chemical composition of HAs would arise following use of the air-drying method. PMID- 17919684 TI - Plio-Pleistocene facies environments from the KBS Member, Koobi Fora Formation: implications for climate controls on the development of lake-margin hominin habitats in the northeast Turkana Basin (northwest Kenya). AB - Climate change is hypothesized as a cause of major events of Plio-Pleistocene East African hominin evolution, but the vertically discontinuous and laterally confined nature of the relevant geological records has led to difficulties with assessing probable links between the two. High-resolution sedimentary sequences from lacustrine settings can provide comprehensive data of environmental changes and detailed correlations with well-established orbital and marine records of climate. Hominin-bearing deposits from Koobi Fora Ridge localities in the northeast Turkana Basin of Kenya are an archive of Plio-Pleistocene lake-margin sedimentation though significant developmental junctures of northern African climates, East African environments, and hominin evolution. This study examines alluvial channel and floodplain, nearshore lacustrine, and offshore lacustrine facies environments for the approximately 136-m-thick KBS Member (Koobi Fora Formation) exposed at the Koobi Fora Ridge. Aspects of the facies environments record information on the changing hydrosedimentary dynamics of the lake margin and give insights into potential climatic controls. Seasonal/yearly climate changes are represented by the varve-like laminations in offshore mudstones and the slickensides, dish-shaped fractures, and other paleosol features overprinted on floodplain strata. Vertical shifts between facies environments, however, are interpreted to indicate lake-level fluctuations deriving from longer-term, dry wet periods in monsoonal rainfall. Recurrence periods for the inferred lake-level changes range from about 10,000 to 50,000 years, and several are consistent with the average estimated timescales of orbital precession ( approximately 20,000 years) and obliquity ( approximately 40,000 years). KBS Member facies environments from the Koobi Fora Ridge document the development of lake-margin hominin habitats in the northeast Turkana Basin. Environmental changes in these habitats may be a result of monsoonal rainfall variations that derive from orbital insolation and/or glacial forcing. PMID- 17919686 TI - Contribution for tier 1 of the ecological risk assessment of Cunha Baixa uranium mine (Central Portugal): I soil chemical characterization. AB - Within the tier 1 of a site specific risk assessment, the pseudo-total concentrations (extracted with aqua regia) and the potential mobile fractions of metals were determined to perform a preliminary evaluation of risks posed by contaminated soils from an abandoned uranium mine (Mangualde, Central Portugal). Considering the mobile fractions of metals, extracted with artificial rain water, aluminium and uranium were the most concerning elements, since their concentrations were above soil quality criteria values (SQGVs) established for both elements. However, according to the evaluation based on potential mobile fractions of elements, rather than on pseudo-total metal concentrations the risks were limited to sites within the exploitation area, where contamination derives mainly from past in-situ leaching activities of pore ore as well as from the deposition of sludge from the effluent pond. The exclusion of other sites under evaluation, from the risk assessment process, requires additional data provided by soil screening ecotoxicological assays. PMID- 17919685 TI - Determination of cadmium in whole blood and scalp hair samples of Pakistani male lung cancer patients by electrothermal atomic absorption spectrometer. AB - A large number of epidemiologic studies have been undertaken to identify potential risk factors for cancer, amongst which the association with cadmium has received considerable attention. There is compelling evidence in support of positive associations between cadmium and risk of lung cancer. In present study we measured the concentration of Cd in whole blood and scalp hair samples of 120 male lung cancer patients (smokers) and 150 controls or referents (smokers and nonsmokers) from different cities of Pakistan. Both referents and patients were of same age group (ranged 40-70 years), socio-economic status, localities and dietary habits. The scalp hair and whole blood samples were oxidized by 65% nitric acid: 30% hydrogen peroxide (2:1) ratio in microwave oven. To check the validity of the proposed method, a conventional wet acid digestion method was used to obtain total Cd concentration in certified samples of human hair BCR 397 and Clincheck control-lyophilized human whole blood. All digests were analyzed for Cd concentration by electrothermal atomic absorption spectrometer (ETAAS). The results of this study showed that the average Cd concentration was higher in the blood and scalp hair of lung cancer patients at different stages as compared to controls (p<001). The smoker referents have high level of Cd in both biological samples as compared to nonsmoker subjects. These results illustrate that the patients who continued smoking after confirmed diagnosis of lung cancer have 34.2-67.26 and 22.4-57.3% more Cd in blood samples and scalp hair than lung cancer patients who cease smoking. This study is compelling evidence in support of positive associations between cadmium, cigarette smoking and lung cancer risk. PMID- 17919687 TI - Effectiveness of household reverse-osmosis systems in a Western U.S. region with high arsenic in groundwater. AB - It is well known to the public in Lahontan Valley in rural Nevada, USA, that local aquifers produce water with varied, but sometimes very high concentrations of arsenic (>4 ppm). As a result, many residents of the area have installed household reverse-osmosis (RO) systems to produce drinking water. We examined performance of RO systems and factors associated with arsenic removal efficiency in 59 households in Lahontan Valley. The sampling results indicated that RO systems removed an average of 80.2% of arsenic from well water. In 18 of the 59 households, arsenic concentrations exceeded 10 ppb in treated water, with a maximum in treated water of 180 ppb. In 3 of the 59 households, RO treatment had little effect on specific conductance, indicating that the RO system was not working properly. Two main factors lead to arsenic levels in treated water exceeding drinking-water standards in the study area. First, arsenic concentrations were high enough in some Lahontan Valley wells that arsenic levels exceeded 10 ppb even though RO treatment removed more than 95% of the arsenic. Second, trivalent As(+3) was the dominant arsenic species in approximately 15% of the wells, which significantly reduced treatment efficiency. Measurements of specific conductance indicated that efficiency in reducing arsenic levels did not always correlate with reductions in total dissolved solids. As a consequence, improvements in taste of the water or simple measurements of specific conductance made by technicians to test RO systems can mislead the public into assuming the water meets safety standards. Actual measurements of treated water are necessary to assure that household RO systems are reducing arsenic concentrations to safe levels, particularly in areas where groundwater has high arsenic concentrations or where As(+3) is the dominant species. PMID- 17919688 TI - Contributions of HERG K+ current to repolarization of the human ventricular action potential. AB - Action potential repolarization in the mammalian heart is governed by interactions of a number of time- and voltage-dependent channel-mediated currents, as well as contributions from the Na+/Ca2+ exchanger and the Na+/K+ pump. Recent work has shown that one of the K+ currents (HERG) which contributes to repolarization in mammalian ventricle is a locus at which a number of point mutations can have significant functional consequences. In addition, the remarkable sensitivity of this K+ channel isoform to inhibition by a variety of pharmacological agents and clinical drugs has resulted in HERG being a major focus for Safety Pharmacology requirements. For these reasons we and others have attempted to define the functional role for HERG-mediated K+ currents in repolarization of the action potential in the human ventricle. Here, we describe and evaluate changes in the formulations for two K+ currents, IK1 and HERG (or IK,r), within the framework of ten Tusscher model of the human ventricular action potential. In this computational study, new mathematical formulations for the two nonlinear K+ conductances, IK1 and HERG, have been developed based upon experimental data obtained from electrophysiological studies of excised human ventricular tissue and/or myocytes. The resulting mathematical model provides much improved simulations of the relative sizes and time courses of the K+ currents which modulate repolarization. Our new formulation represents an important first step in defining the mechanism(s) of repolarization of the membrane action potential in the human ventricle. Our overall goal is to understand the genesis of the T-wave of the human electrocardiogram. PMID- 17919689 TI - Theoretical analysis of the magnetocardiographic pattern for reentry wave propagation in a three-dimensional human heart model. AB - We present a computational study of reentry wave propagation using electrophysiological models of human cardiac cells and the associated magnetic field map of a human heart. We examined the details of magnetic field variation and related physiological parameters for reentry waves in two-dimensional (2-D) human atrial tissue and a three-dimensional (3-D) human ventricle model. A 3-D mesh system representing the human ventricle was reconstructed from the surface geometry of a human heart. We used existing human cardiac cell models to simulate action potential (AP) propagation in atrial tissue and 3-D ventricular geometry, and a finite element method and the Galerkin approximation to discretize the 3-D domain spatially. The reentry wave was generated using an S1-S2 protocol. The calculations of the magnetic field pattern assumed a horizontally layered conductor for reentry wave propagation in the 3-D ventricle. We also compared the AP and magnetocardiograph (MCG) magnitudes during reentry wave propagation to those during normal wave propagation. The temporal changes in the reentry wave motion and magnetic field map patterns were also analyzed using two well-known MCG parameters: the current dipole direction and strength. The current vector in a reentry wave forms a rotating spiral. We delineated the magnetic field using the changes in the vector angle during a reentry wave, demonstrating that the MCG pattern can be helpful for theoretical analysis of reentry waves. PMID- 17919690 TI - Midkine promoter-based conditionally replicative adenovirus for targeting midkine expressing human bladder cancer model. AB - OBJECTIVES: To develop a novel therapeutic strategy against human bladder cancer using Ad-MK-E1a-a midkine (MK) promoter-regulated, conditionally replicating, adenovirus. METHODS: We tested several human cancer cell lines in vitro, including those of bladder cancer (KK47, 5637, and T24), lung cancer (A549), and head and neck cancer (H891). In each cell line, we examined MK mRNA expression by TaqMan real-time quantitative polymerase chain reaction, MK promoter activity, after plasmid transfection, using a luciferase assay, and the transduction efficiency by co-transfection with the cytomegalovirus-beta-gal plasmid. In these cells, we assessed the cell type-specific replication of Ad-MK-E1a virus by measuring the E1a DNA copy number by real-time polymerase chain reaction and the cell growth inhibition due to this virus using the Alamar blue assay. In animal studies, nude mice were subcutaneously inoculated with KK47 cells and later intratumorally injected with phosphate-buffered saline or Ad5-CMV-LacZ or Ad-MK E1a. RESULTS: The MK mRNA expression level and MK promoter-driven luciferase activity were relatively greater and markedly increased, respectively, in the 5637, A549, and KK47 cells than in the T24 and H891 cells. After Ad-MK-E1a infection, the E1a DNA copy number increased more significantly in the KK47, 5637, and A549 cells than in the T24 and H891 cells. At a multiplicity of infection of 0.01, Ad-MK-E1a significantly inhibited KK47 and 5637 cell growth. In vivo, Ad-MK-E1a injection markedly inhibited KK47 tumor growth. CONCLUSIONS: We have demonstrated the antitumor effect of Ad-MK-E1a in a human bladder cancer model overexpressing MK mRNA. PMID- 17919691 TI - Routine stenting reduces urologic complications as compared with stenting "on demand" in adult kidney transplantation. AB - OBJECTIVES: To examine the impact of the chosen surgical technique and of systematic versus "on-demand" placement of a primary stent on the incidence of urologic complications in adult kidney transplantation. METHODS: Data of 497 consecutive patients undergoing kidney transplantation at a single center were retrospectively analyzed with respect to urologic complications. Three different surgical strategies for the ureteroneocystostomy were compared: (1) transvesical anastomosis with stenting "on demand," (2) extravesical anastomosis with stenting "on demand," and (3) extravesical anastomosis with routine stenting. Nine parameters were evaluated by logistic regression for a possible contribution to the development of urologic complications. RESULTS: Routine placement of a stent significantly reduced the number of urologic complications compared with both transvesical or extravesical anastomoses with stenting "on demand" (20.8% in transvesical "on demand," 17.9% in extravesical "on demand," and 5.8% in extravesical "routine"). Logistic regression analysis revealed that routine stenting versus stenting "on demand" (P = 0.001) and living donor transplantation (P = 0.038) are two independent factors associated with a significantly lower incidence of urologic complications. Although routine stenting was not associated with an increased incidence of urinary tract infections, female gender was the only independent factor associated with this complication (P = 0.001). CONCLUSIONS: Routine stenting of the ureteroneocystostomy is superior to stenting "on demand" in adult kidney transplantation, suggesting that the intraoperative decision of whether to stent is insufficient to avoid urologic complications. PMID- 17919692 TI - Identification of early p53 mutations in clam ileocystoplasties using restriction site mutation assay. AB - OBJECTIVES: Because a risk of cancer arising in enterocystoplasties exists, it is necessary to identify which patients are most at risk of tumor formation. The aim of this study was to determine whether rare mutated p53 sequences were more common at the enterovesical anastomosis than in the bladder remnant in patients with a clam ileocystoplasty using the restriction site mutation (RSM) assay. METHODS: DNA was extracted from endoscopic biopsies obtained from the ileovesical anastomosis and native bladder remnant (control specimens) of 38 patients with a clam ileocystoplasty. The RSM assay was used to study five known hotspots for mutations of the p53 gene using the restriction enzymes Hha I (codon 175), Taq I (codon 213), Hae III (codon 249/250), and Msp I (codons 248 and 282). The mutational events of p53 were confirmed by sequencing the undigested mutated polymerase chain reaction products identified by RSM analysis. RESULTS: We found p53 mutations at the ileovesical anastomosis in 7 of the 38 patients. The mutations were observed at codon 213 (n = 1), codon 248 (n = 3), and codon 250 (n = 3). No p53 mutations were detected in any control specimen. CONCLUSIONS: The ileovesical anastomosis is genetically unstable in patients with a clam ileocystoplasty. The p53 mutations identified by the RSM assay at the enterovesical anastomosis could possibly be used as markers of genetic instability to identify patients at risk of developing a tumor. Prospective, randomized longitudinal studies are required to substantiate this hypothesis. PMID- 17919694 TI - Short-term health outcome differences between robotic and conventional radical prostatectomy. AB - OBJECTIVES: To prospectively compare the relevant in-hospital and postdischarge short-term health outcomes in a contemporary group of patients undergoing either robotic or conventional radical prostatectomy. METHODS: A total of 117 robotic and 89 conventional radical prostatectomy patients participated in a prospective study evaluating short-term postoperative recovery. A validated short-term health outcomes questionnaire was self-administered at 2 and 6 weeks postoperatively. All patients were given the same preoperative and postoperative instructions. RESULTS: The significant differences in short-term health outcome measures between the two groups were that robotic patients had less mean narcotic use during the first 24 hours of admission (32 versus 52 mg morphine sulfate equivalents, P = 0.001) and a shorter mean length of hospital stay (1.2 versus 1.3 days, P = 0.048). No difference was found between the robotic and conventional radical prostatectomy groups regarding the median time to normal activity, 100% activity, and time to driving (9 and 8, 21 and 28, and 13 and 14 days, respectively). No difference was found in the postdischarge pain levels at 2 and 6 weeks, as reflected in the equivalent time of narcotic use between the two groups (10.6 and 9 days for the robotic and conventional radical prostatectomy groups, respectively). CONCLUSIONS: The results of this prospective study have shown that both robotic and conventional radical prostatectomy provide comparable short-term postdischarge recovery, including time to normal and full activity, driving, and postdischarge narcotic use. PMID- 17919695 TI - Mechanisms of hemostatic failure during laparoscopic nephrectomy: review of Food and Drug Administration database. AB - OBJECTIVES: To compare the complications with endovascular stapling devices, nonlocking titanium clips, and nonabsorbable polymer ligating (Hem-o-lok) clips during laparoscopic nephrectomy. METHODS: The Food and Drug Administration Manufacturer and User Facility Device Experience Database was retrospectively reviewed for reports dated from January 1992 to March 2006 using the key words "nephrectomy" and "kidney." All episodes of pure and hand-assisted laparoscopic nephrectomy were evaluated. RESULTS: Of 2172 total nephrectomy or kidney-related reports, 352 reported failure using laparoscopic hemostatic devices to secure the renal vasculature, and 223 complications (63%) resulted during the use of endovascular stapling devices, 111 (33%) from nonlocking titanium clips and 18 (5%) from locking clips. The leading causes of failure reported in stapling devices were staple line malformation (47%) and locking up (29%). In titanium clips, jamming/feeding difficulties (27%) and trouble closing or "scissoring" clips (26%) were the most common. In locking clips, dislodgement (44%) was most frequently reported. Three, one, and three deaths were reported after the use of the stapling device, titanium clip, and locking clip device, respectively. CONCLUSIONS: All three methods used to secure the renal hilum in laparoscopic nephrectomy can result in malfunction. Because the overall denominator of use is not known, it would be inappropriate to conclude that one device is safer than another. When they occurred, these device malfunctions were potentially serious. Knowledge of the possible mechanisms of failure seen with each device could allow surgeons to anticipate potential complications and, therefore, perform laparoscopic surgery more safely. PMID- 17919696 TI - Lemonade therapy increases urinary citrate and urine volumes in patients with recurrent calcium oxalate stone formation. AB - OBJECTIVES: Potassium citrate is prescribed to patients with calcium oxalate (CaOx) stone formation to increase urinary citrate and pH, thus reducing CaOx crystal formation. Lemonade therapy (LT) might also increase urinary citrate and the total urine volume. We compared the effects of LT alone (group 1, n = 63) and potassium citrate plus LT (group 2, n = 37) in patients with CaOx stone formation on the urinary citrate and total urine volume to determine the efficacy of LT. METHODS: Adult patients with CaOx stone formation and three or more clinic visits from 1996 to 2005 and three or more UroRisk profiles were included in our retrospective analysis. RESULTS: Urinary citrate increased maximally by 203 and 346 mg/day for groups 1 and 2, respectively. The maximal total urine volume increase was 763 and 860 mL/day for groups 1 and 2, respectively. The urinary citrate and total urine volume increased sooner during follow-up for group 1. By the last clinic visit, the urinary citrate and total urine volume had decreased in both groups. However, group 1 sustained a greater total urine volume than did group 2 (2.35 +/- 0.10 standard error versus 2.17 +/- 0.12 L/day). Urinary citrate was greater in group 1 (765 +/- 56 standard error versus 548 +/- 56 mg/day for group 2), but the change from baseline to the last visit was significant (P = 0.008) only in group 2. CONCLUSIONS: LT resulted in favorable changes in urinary citrate and total urine volume in our series. Potassium citrate with LT was more effective than LT alone at increasing urinary citrate. Because maximal changes for urinary citrate and total urine volume were achieved earlier in follow-up, individualized encouragement and motivation should be provided to patients at each visit for sustained prevention. PMID- 17919697 TI - Efficacy of transcutaneous electrical nerve stimulation in children with overactive bladder refractory to pharmacotherapy. AB - OBJECTIVES: To examine the outcome of transcutaneous electrical nerve stimulation (TENS) use in children with overactive bladder refractory to behavioral and anticholinergic therapy. METHODS: We reviewed the data of 18 children (13 girls and 5 boys; mean age 9 years) with overactive bladder refractory to standard therapy who underwent at-home TENS treatment twice daily for 20 minutes. Symptoms and objective measures (noninvasive uroflowmetry and postvoid residual urine volume) were noted at baseline and after treatment. RESULTS: Of the 18 children, 15 had pretreatment incontinence and 3 had only increased urgency/frequency. The incontinence group averaged 3.2 +/- 2.1 daytime accidents. The mean length of TENS use was 8 +/- 7 months, and the mean follow-up after starting TENS was 13 +/ 9 months. Of the 15 patients with incontinence, 2 became dry (13%), 9 were significantly improved (60%), and 4 reported no improvement (27%). Of 12 children with marked urinary frequency, 8 had significant symptom improvement. Noninvasive uroflow and postvoid residual urine volume measurements were not significantly different statistically before and after treatment. Pretreatment patient sex, age, and frequency of wetting were not predictive of the outcome. A post-TENS parabolic uroflow curve showed a statistically significant correlation with patients who became dry or improved (P = 0.018). CONCLUSIONS: The results of our study have indicated that TENS use in children with incontinence refractory to pharmacotherapy is safe and well tolerated. The encouraging results of this treatment modality in the refractory patient population warrant additional study of the pediatric overactive bladder. PMID- 17919693 TI - Relationship of lifestyle and clinical factors to lower urinary tract symptoms: results from Boston Area Community Health survey. AB - OBJECTIVES: Because lifestyle factors and comorbidities can influence lower urinary tract symptoms (LUTS) by sex and race/ethnicity differently, we investigated these associations in the Boston Area Community Health survey. METHODS: Using a multistage stratified cluster random sample, 5506 adults aged 30 to 79 years were enrolled (2301 men, 3205 women, 1770 blacks, 1877 Hispanics, and 1859 whites). Adiposity, lifestyle factors, comorbidities (cardiovascular diseases, diabetes, high blood pressure, high cholesterol, depressive symptoms, previous urinary tract infections) were considered in predicting the odds of LUTS (American Urological Association Symptom Index of 8 or greater) by sex and race/ethnicity. RESULTS: The prevalence of LUTS was 18.7%, with similar rates by sex (men 18.7%, women 18.6%) and race/ethnicity (black 19.3%, Hispanic 16.2%, white 18.9%); however, the prevalence did increase substantially with age. Depressive symptoms were associated with increased odds of LUTS across all sex and racial/ethnic groups. The overall odds ratio was 2.4 (95% confidence interval 1.9 to 3.2, P <0.001). Age increased the odds of LUTS among all groups. Physical activity decreased the odds of LUTS, particularly in women (odds ratio 0.4, 95% confidence interval 0.2 to 0.7, P = 0.003, comparing high and low activity). Cardiovascular diseases and previous urinary tract infections increased the odds of LUTS overall (odds ratio 1.6, 95% confidence interval 1.2 to 2.1, P = 0.004; and odds ratio 1.9, 95% confidence interval 1.4 to 2.4, P <0.001, respectively) and for most groups. CONCLUSIONS: The results of this study have shown that the lifestyle and clinical factors associated with LUTS are similar by sex and race/ethnicity. PMID- 17919698 TI - Evaluation of practice efficiency with a novel sheathed flexible cystoscope: a randomized controlled trial. AB - OBJECTIVES: The processing of standard flexible cystoscopes is time-consuming and potentially hazardous to healthcare staff. We evaluated a novel flexible cystoscope system containing a disposable sheath, which obviates the need for chemical processing. METHODS: A total of 100 patients undergoing office cystoscopy were randomized into a "sheath" or a control group (n = 50 for each group). Disposable Slide-On sheaths and the CST-2000 Flexible Cystoscope System were used in the former group. A standard 16F flexible cystoscope, manually cleaned and disinfected with Cidex OPA or Rapicide, was used in the control group and the preparation times were recorded. Questionnaires to assess cystoscope setup, handling, optical quality, and patient comfort were completed by the physician, nursing staff, and patients. RESULTS: Data were available for 97 patients. The mean total time required for preparation of the cystoscopes was 10.7, 14.7, and 42.2 minutes for the sheath (n = 49), Rapicide (n = 21), and Cidex OPA (n = 27), respectively (P <0.01 for all comparisons). The control group scored better than the sheath group with regard to ease of cystoscope insertion (9.1 versus 7.7), cystoscope handling (8.8 versus 8.1), and irrigation setup (8.8 versus 8.2; all P < or = 0.01). No statistically significant difference was found between groups for procedure time, optical quality, or patient comfort, either during or after the procedure. CONCLUSIONS: The EndoSheath System can save 4 to 31 minutes of processing time while avoiding exposure to irritants found in conventional soaking methods. Although maneuvering of the sheathed cystoscope was slightly more difficult, the optical quality and patient comfort were similar to those of standard flexible cystoscopy. PMID- 17919699 TI - Salvage laparoscopic extraperitoneal radical prostatectomy after failed high intensity focused ultrasound and radiotherapy for localized prostate cancer. AB - OBJECTIVES: To investigate the feasibility and efficacy of salvage endoscopic extraperitoneal radical prostatectomy (EERPE) in cases of recurrent prostate cancer after high-intensity focused ultrasound therapy (HIFU) or radiotherapy. METHODS: Nine patients underwent salvage EERPE with curative intent for biopsy proven, locally recurrent prostate cancer. Of these 9 patients, 3 had previously undergone HIFU and 6 radiotherapy. Perioperative parameters (operation time, estimated blood loss, conversion to open surgery rate, transfusion rate, transurethral catheter time), functional outcome, and short-term oncologic outcome were reviewed. RESULTS: Mean patient age was 63.3 years (range 48 to 74 years). Mean preoperative PSA value was 12.64 ng/mL and mean prostate weight was 49.2 g. Mean blood loss was 238 mL. There was no need for conversion to open surgery or transfusion. Mean operation time was 148 minutes, and mean total transurethral catheter time was 6 days. No intraoperative complications were reported. There was no clear difference in operation difficulty between the post HIFU and postradiotherapy EERPE. After a mean follow-up of 17 months, 7 patients were completely continent, and 2 needed 1 to 2 pads per day. Three patients were potent before the surgical treatment, but no patient reported potency postoperatively. In 1 patient a PSA relapse (1.20 ng/mL) was recognized 12 months postoperatively. CONCLUSIONS: Salvage EERPE after failed HIFU and radiation therapy is a safe and efficient method to treat locally recurrent prostate cancer. Short-term oncologic and functional outcomes are promising, but further study should be made on the long-term oncologic outcomes of this technique. PMID- 17919701 TI - Retroperitoneal vascular leakage in ovarian cancer is associated with increased VEGF. PMID- 17919700 TI - Reconstruction of the pelvic floor and perineum with human acellular dermal matrix and thigh flaps following pelvic exenteration. AB - BACKGROUND: Patients who undergo pelvic floor resection as treatment for recurrent cancer following radiation therapy have increased rates of complications, particularly if permanent prosthetic mesh is used for reconstruction. Human acellular dermal matrix (HADM), commonly used for reconstruction in other torso locations, is associated with lower rates of complications (including infection, adhesions and cutaneous exposure) than synthetic mesh. We describe an effective technique to reconstruct the pelvic floor and perineum with HADM and thigh-based flaps following pelvic exenteration and radical vulvectomy. CASE: A 75-year-old woman underwent radical resection of the pelvic floor and perineum to treat recurrent vulvar squamous cell carcinoma and osteoradionecrosis. The pelvic floor and perineal soft tissue defect were reconstructed with HADM (AlloDerm; LifeCell Corporation, Branchburg, NJ) and bilateral, thigh-based tissue flaps, respectively. Despite a large resection, previous irradiation therapy and bacterial contamination the wounds healed without complications. CONCLUSION: Reconstruction of pelvic floor defects using HADM is an option when wound conditions are unfavorable for the use of permanent prosthetic meshes. PMID- 17919702 TI - Innate immunomodulatory effects of cereal grains through induction of IL-10. AB - BACKGROUND: Interactions between the gastrointestinal immune system and the luminal environment play critical roles in maintaining immune homeostasis and in diseases such as inflammatory bowel disease. Although immunomodulation by microbial factors has been studied extensively, little attention has been given to the potential immunomodulatory effects of ingested foods. OBJECTIVE: We characterized the effects of cereal grains on the immune response in human subjects and investigated the mechanism. METHODS: PBMCs from healthy individuals were incubated with cereal grain extracts, and cytokine levels in cell-free supernatants were measured. The cellular source of IL-10 and the role of monocytes were investigated by means of flow cytometry and cell depletion/reconstitution experiments. RESULTS: Extracts of cereal grains, including rice and wheat, induced marked IL-10 production from PBMCs. Intracellular cytokine staining and cell-depletion experiments showed that CD14+ monocytes produced IL-10. Importantly, when PBMCs were stimulated with concanavalin A, cereal grains concentration-dependently inhibited their production of IL-5, IL-13, and IFN-gamma; neutralizing IL-10 or removing the monocytes abrogated this inhibitory effect. This cereal grain-induced IL-10 response was polymyxin B sensitive, heat resistant, and inhibited by blocking the Toll-like receptor 4. CONCLUSION: Cereal grains have strong innate immunomodulatory effects by inducing marked production of IL-10 from CD14+ monocytes in vitro. LPS or LPS-mimicking activity in cereal grains might be responsible. The potential immunomodulatory effects of cereal grains need further study in vivo. PMID- 17919703 TI - The GILL study: glycerin-induced local reactions in immunotherapy. AB - BACKGROUND: The mechanism of local reactions is not well defined. Glycerin, an excellent preservative used commonly in immunotherapy extracts, is a recognized irritant. OBJECTIVE: This study was undertaken to examine whether higher glycerin concentration in immunotherapy extracts is associated with an increase in local reaction rates during immunotherapy. METHODS: A retrospective analysis of electronic immunotherapy records over a 12-month period was performed from a single site. A small local reaction was defined as induration and/or erythema at the injection site smaller than or equal to the size of the patient's palm. A large local reaction was defined as a reaction larger than the patient's palm. RESULTS: Over the 12-month period, 360 patients received a total of 9678 immunotherapy injections. For all injections, the total local reaction rate was 16.3% (1574/9678), the small local reaction rate was 15.9% (1536/9678), and the large local reaction rate was 0.4% (38/9678). For aeroallergens, small local reaction rates increased significantly with increasing allergen concentrations, from 7.3% (1:1000 vol/vol) to 23.0% (1:1 vol/vol; P < .001). The small local reaction rate was higher with increasing allergen content but not higher glycerin concentration. Large local reactions were infrequent and did not significantly increase with allergen or glycerin concentration. CONCLUSIONS: Small local, but not large local, reaction rates increase with higher allergen concentration, number, and volume. Higher glycerin concentrations (even 50%) are not associated with significantly higher small or large local reaction rates. PMID- 17919705 TI - Is vitamin D deficiency to blame for the asthma epidemic? AB - In the 1960s, the prevalence of asthma and allergic diseases began to increase worldwide. Currently, the burden of the disease is more than 300 million people affected. We hypothesize that as populations grow more prosperous, more time is spent indoors, and there is less exposure to sunlight, leading to decreased cutaneous vitamin D production. Coupled with inadequate intake from foods and supplements, this then leads to vitamin D deficiency, particularly in pregnant women, resulting in more asthma and allergy in their offspring. Vitamin D has been linked to immune system and lung development in utero, and our epidemiologic studies show that higher vitamin D intake by pregnant mothers reduces asthma risk by as much as 40% in children 3 to 5 years old. Vitamin D deficiency has been associated with obesity, African American race (particularly in urban, inner-city settings), and recent immigrants to westernized countries, thus reflecting the epidemiologic patterns observed in the asthma epidemic. Providing adequate vitamin D supplementation in pregnancy may lead to significant decreases in asthma incidence in young children. PMID- 17919706 TI - Atopy in early life and effect of a primary prevention program for asthma in a high-risk cohort. PMID- 17919707 TI - Expression of IL-9 receptor alpha chain on human germinal center B cells modulates IgE secretion. AB - BACKGROUND: IL-9 has been shown to affect the differentiation pathway of different cell types. However, its potential role in the maturation pathway of antigen-driven B-cell differentiation and its functional effects remain unknown. OBJECTIVE: To characterize IL-9 receptor alpha chain (IL-9R alpha) expression on human tonsillar B cells at different maturational stages, and to assess its effect on IgE production. METHODS: Freshly purified human tonsillar B cells were fractionated into 3 populations: low-density (LD), medium-density, and high density cells. Expression levels of IL-9R alpha were determined by using immunohistochemistry and flow cytometry. IL-9R alpha(high)-expressing cells were stimulated with IL-9 in the presence or absence of IL-4, and IgE release was measured by ELISA. RESULTS: IL-9R alpha was expressed on human LD tonsillar B cells, with an ability to transduce signals through activation of signal transducer and activator of transcription 3 and 5. Although IL-9 was unable to induce IgE secretion by itself, it potentiated IL-4-mediated IgE production from LD cells. Moreover, increased IgE was paralleled by an upregulation of IL-9R alpha and CD27, with the latter a memory B-cell marker implicated in increased IgE secretion. CONCLUSION: These results highlight a crucial role for IL-9 in modulating T-cell-dependent B-cell differentiation and establish a new paradigm for understanding the synergistic role of T(H)2 cytokines and their modulatory effect on B-cell maturation and IgE production. CLINICAL IMPLICATIONS: IL-9 appears to be involved in memory B-cell differentiation and T(H)2-mediated allergic diseases such as asthma. PMID- 17919708 TI - Exogenous vitamin D might contribute to geographic variations in epinephrine prescriptions. PMID- 17919709 TI - A polymorphism in CD14 modifies the effect of farm milk consumption on allergic diseases and CD14 gene expression. AB - BACKGROUND: Consumption of farm milk in early life is associated with less asthma and allergies. OBJECTIVE: We hypothesized that genetic variation in the innate immunity receptor CD14 might modify the association between farm milk consumption and asthma and atopy. METHODS: Questionnaire data, serum IgE levels, and genotypes for 4 single nucleotide polymorphisms in CD14 were assessed in farmers' and nonfarmers' children from 2 European populations (Allergy and Endotoxin study, n = 576; Prevention of Allergy Risk factors for Sensitization in children related to Farming and Anthroposophic Lifestyle study, n = 1539). In a subsample (n = 222) CD14 gene expression was measured in peripheral blood leukocytes. The effects of farm milk and CD14 genotypes on asthma, allergies, and CD14 expression and their interactions were investigated. RESULTS: We found a significant interaction between genetic variation in CD14/-1721 and farm milk consumption. Adjusted odds ratios for the association between farm milk and asthma varied between the genotypes: AA, 0.18 (95% CI, 0.07-0.47); AG, 0.47 (95% CI, 0.26 0.86); and GG, 0.98 (95% CI, 0.46-2.08). Similar patterns were observed for symptoms of allergic rhinoconjunctivitis and pollen sensitization. CD14/-1721 also modified the association between farm milk and CD14 gene expression (adjusted geometric means ratios: AA, 1.61 (95% CI, 0.98-2.66); AG, 1.11 (95% CI, 0.71-1.72); and GG, 0.76 (95% CI, 0.39-1.48). CONCLUSION: The protective effect of farm milk consumption on allergic diseases is stronger in children carrying the A allele in CD14/-1721 than in children homozygous for the G allele. This might be mediated through farm milk-induced upregulated CD14 gene expression. CLINICAL IMPLICATIONS: Our results support the hypothesis that the inverse association between farm milk consumption and allergic diseases is mediated by CD14-activated innate immune mechanisms. PMID- 17919712 TI - Anaphylactic shock after first exposure to gadoterate meglumine: two case reports documented by positive allergy assessment. PMID- 17919713 TI - A birth of inactivity? A review of physical activity and parenthood. AB - OBJECTIVE: To review the existing research on physical activity and parenthood in order to establish direction for future research. METHOD: Articles were limited to English peer-reviewed journals, published from 1989 to 2007. Major findings from 25 independent samples were summarized based on common subtopics of: physical activity of parents compared to non-parents, physical activity barriers, employment and marital status, number of children, and theory-based work applied to parents. RESULTS: Parenthood and physical activity involvement showed a negative relationship (meta-analytic d=0.41 to 0.48, correcting for sampling error) when compared to non-parents. Mothers were generally less active than fathers. Associations were found between specific barriers and parental physical activity, but the relationship between physical activity and marital/employment status as well as number/age of children was inconsistent. Finally, the use of theoretical models applied to understanding early family development and physical activity has been limited. CONCLUSIONS: Parents with dependent children are clearly more inactive than non-parents and the topic has received disproportionably scant research considering the size of the effect. Current research has largely been focused on mothers, and has relied heavily on cross sectional designs and self-report measures. Future work should focus on longitudinal designs across family development, gender and role interactions, and include social ecological frameworks and objective physical activity measurement. PMID- 17919711 TI - Altered phosphorylated signal transducer and activator of transcription profile of CD4+CD161+ T cells in asthma: modulation by allergic status and oral corticosteroids. AB - BACKGROUND: Asthma is a complex immunologic disorder linked to altered cytokine signaling. OBJECTIVE: We tested whether asthmatic patients showed any change in cytokine-dependent signal transducer and activator of transcription (STAT) levels, focusing on the central/effector-memory CD4(+)CD161(+) subset, which represents 15% to 25% of circulating T cells. METHODS: We quantified intracellular levels of active phosphorylated STAT (phospho-STAT) 1, 3, 5, and 6 by means of flow cytometry, without any activation or expansion. RESULTS: Baseline phospho-STAT1 and phospho-STAT6 levels were increased in CD4(+)CD161(+) T cells from asthmatic patients compared with those from healthy control subjects (by 10- and 8-fold, respectively). This asthma-associated alteration was both subset specific because no change was seen in CD4(+)CD161(-)CD25(+) (regulatory T cells) and CD4(+)CD161(-)CD25(-) subsets and isoform specific because phospho STAT5 and phospho-STAT3 levels were unchanged. Among asthmatic patients, phospho STAT1 and phospho-STAT6 levels correlated negatively with each other, suggesting antagonistic regulation. Oral corticosteroid (OCS) treatment significantly decreased phospho-STAT6 and IL-4 levels but not phospho-STAT1 levels. Disease parameters showing significant correlations with phospho-STAT1, phospho-STAT6, or both included age at onset, plasma IgE levels, and levels of the T(H)2 cytokines IL-4 and IL-10 and the T(H)1 cytokine IL-2. Overall, combined phospho-STAT1 and phospho-STAT6 measurements showed excellent predictive value for identifying (1) asthmatic patients versus healthy control subjects, (2) allergic versus nonallergic asthmatic patients, and (3) asthmatic patients taking versus those not taking OCSs. CONCLUSION: Baseline changes in phospho-STAT1 and phospho-STAT6 levels in blood CD4(+)CD161(+) T cells identify asthmatic patients and mirror their allergic status and response to OCSs. CLINICAL IMPLICATIONS: These results confirm the pathologic importance of activated STAT1 and STAT6 in asthma and suggest their potential use as clinical biomarkers. PMID- 17919714 TI - Early detection of skin cancer as public health policy: comparison of campaign and routine activity. AB - OBJECTIVE: The objective of this study was to evaluate the early detection of skin cancer, with emphasis on comparison of campaign (short high-intensity periods) and routine (continuous low-intensity) activity. METHODS: The study population consisted of 5903 campaign and 4284 routine attenders visiting a nurse at the Pirkanmaa Cancer Society between January 1, 1991, and December 31, 2000. Skin cancers were identified from the Finnish Cancer Registry. The performance of the program was evaluated in terms of sensitivity, specificity, positive and negative predictive values. RESULTS: A single lesion was more frequently examined during campaigns whereas a partial or total body were examined less frequently (p<0.001). Attenders received referral for removal of a lesion more frequently in routine activity (52% vs. 20% p<0.001), regardless of extent of the examination. The cumulative incidence of skin cancer within 24 months was 3.2% for routine and 1.6% for campaign attenders (p<0.001). Sensitivity was higher (82% vs. 59%, p<0.001), while specificity was lower (49% vs. 79%, p<0.001) for routine activity. CONCLUSIONS: Even though neither approach appears optimal, scheduled appointments with adequate time allocation per subject provided a lower threshold for detection and a higher yield of skin cancers compared to high-intensity campaigns. PMID- 17919715 TI - Dynamics of sperm DNA fragmentation in domestic animals II. The stallion. AB - The mixed success of equine artificial insemination programs using chilled and frozen-thawed semen is most likely associated with the variable response of the sperm cell to the preservation process and the fact that stallions are not selected on the basis of reproductive performance. We propose that the traditional indicators of sperm viability do not fully account for male factor infertility in the stallion and that knowledge of sperm DNA damage in the original semen sample and during semen processing may provide a more informed explanation of an individual stallion's reproductive potential. This study reports on the validation of a sperm DNA fragmentation test based on the sperm chromatin dispersion test (SCD) for stallion spermatozoa and on its application to semen that was chilled (4 degrees C; n=10) or frozen-thawed (n=13). Semen samples were collected by artificial vagina and the proportion of sperm with fragmented DNA determined. Seminal plasma was then removed by centrifugation and the sperm pellet re-suspended in commercial extenders prior to being chilled or cryopreserved using standard industry protocols. Chilled semen was cooled slowly to 4 degrees C and stored for 1h before commencing the analysis; cryopreserved semen was thawed and immediately analyzed. Following chilling or cryopreservation, the semen samples were incubated at 37 degrees C and analyzed for SCD after 0, 4, 6, 24 and 48 h storage. The results of this investigation revealed that there was no significant difference in the sperm DNA fragmentation index (sDFI) of sperm evaluated initially after collection compared to those tested immediately after chilling or cryopreservation. However, within 1h of incubation at 37 degrees C, both chilled and frozen-thawed spermatozoa showed a significant increase in the proportion of sDFI; after 6h the sDFI had increased to over 50% and by 48 h, almost 100% of the sperm showed DNA damage. While the sDFI of individual stallions at equivalent times of incubation was variable, an analysis of the rate of change of sDFI revealed no difference between stallions or the way in which the semen was preserved. In terms of sperm DNA fragmentation dynamics, the highest intensity of sperm DNA damage occurred in the first 6h of incubation. We suggest that the SCD test can be used as a routine assessment tool for the development and refinement of preservation protocols designed to reduce stallion sperm DNA damage. PMID- 17919716 TI - Identification of differentially expressed genes from the cross-subfamily cloned embryos derived from zebrafish nuclei and rare minnow enucleated eggs. AB - Cross-species nuclear transfer (NT) has been used to retain the genetic viability of a species near extinction. However, unlike intra-species NT, most embryos produced by cross-species NT were unable to develop to later stages due to incompatible nucleo-cytoplasmic interactions between the donor nuclei and the recipient cytoplasm from different species. To study the early nucleo-cytoplasmic interaction in cross-species NT, two laboratory fish species (zebrafish and rare minnow) from different subfamilies were used to generate cross-subfamily NT embryos in the present study. Suppression subtractive hybridization (SSH) was performed to screen out differentially expressed genes from the forward and reverse subtractive cDNA libraries. After dot blot and real-time PCR analysis, 80 of 500 randomly selective sequences were proven to be differentially expressed in the cloned embryos. Among them, 45 sequences shared high homology with 28 zebrafish known genes, and 35 sequences were corresponding to 22 novel expressed sequence tags (ESTs). Based on functional clustering and literature mining analysis, up- and down-regulated genes in the cross-subfamily cloned embryos were mostly relevant to transcription and translation initiation, cell cycle regulation, protein binding, etc. To our knowledge, this is the first report on the determination of genes involved in the early development of cross-species NT embryos of fish. PMID- 17919717 TI - HPV testing for primary cervical cancer screening. PMID- 17919718 TI - Human papillomavirus DNA testing for the detection of cervical intraepithelial neoplasia grade 3 and cancer: 5-year follow-up of a randomised controlled implementation trial. AB - BACKGROUND: Tests for the DNA of high-risk types of human papillomavirus (HPV) have a higher sensitivity for cervical intraepithelial neoplasia grade 3 or worse (CIN3+) than does cytological testing, but the necessity of such testing in cervical screening has been debated. Our aim was to determine whether the effectiveness of cervical screening improves when HPV DNA testing is implemented. METHODS: Women aged 29-56 years who were participating in the regular cervical screening programme in the Netherlands were randomly assigned to combined cytological and HPV DNA testing or to conventional cytological testing only. After 5 years, combined cytological and HPV DNA testing were done in both groups. The primary outcome measure was the number of CIN3+ lesions detected. Analyses were done by intention to treat. This trial is registered as an International Standard Randomised Controlled Trial, number ISRCTN20781131. FINDINGS: 8575 women in the intervention group and 8580 in the control group were recruited, followed up for sufficient time (> or =6.5 years), and met eligibility criteria for our analyses. More CIN3+ lesions were detected at baseline in the intervention group than in the control group (68/8575 vs 40/8580, 70% increase, 95% CI 15-151; p=0.007). The number of CIN3+ lesions detected in the subsequent round was lower in the intervention group than in the control group (24/8413 vs 54/8456, 55% decrease, 95% CI 28-72; p=0.001). The number of CIN3+ lesions over the two rounds did not differ between groups. INTERPRETATION: The implementation of HPV DNA testing in cervical screening leads to earlier detection of CIN3+ lesions. Earlier detection of such lesions could permit an extension of the screening interval. PMID- 17919719 TI - Ovotransferrin possesses SOD-like superoxide anion scavenging activity that is promoted by copper and manganese binding. AB - The embryo of oviparous species is confronted by a highly oxidative stress generating as it grows and must rely on effective antioxidant system for protection. Proteins of avian egg albumin have been suggested to play the major redox-modulatory role during embryo development. Recently, we found that ovotransferrin (OTf) undergoes distinct thiol-linked self-cleavage in a redox dependent process. In this study, we explore that OTf is SOD mimic protein with a potent superoxide anion (O(2)(-)) scavenging activity. The O(2)(-) scavenging activity was investigated using the natural xanthine/xanthine oxidase (X/XOD) coupling system. OTf exhibited O(2)(-) scavenging activity in a dose-dependent manner and showed remarkably higher scavenging activity than the known antioxidants, ascorbate or serum albumin. The isolated half-molecules of OTf exhibited higher scavenging activity than the intact molecule, whereas the N-lobe showed much greater activity. OTf dramatically quenched the O(2)(-) flux but had no effect on the urate production in the X/XOD system, indicating its unique specificity to scavenge O(2)(-) but not oxidase inhibition. Strikingly, metal bound OTf exhibited greater O(2)(-) dismutation capacity than the apo-protein, ranging from moderate (Zn(2+)-OTf and Fe(2+)-OTf) to high (Mn(2+)-OTf and Cu(2+) OTf) activity with the Cu(2+)-OTf being the most potent scavenger. In a highly sensitive fluorogenic assay, the metal-bound OTf exhibited significant increase in the rate of H(2)O(2) production in the X/XOD reaction than the apo-OTf, providing evidence that Zn(2+)-, Mn(2+)- and Cu(2+)-OTf possess SOD mimic activity. This finding is the first to describe that OTf is an O(2)(-) scavenging molecule, providing insight into its novel SOD-like biological function, and heralding a fascinating opportunity for its potential candidacy as antioxidant drug. PMID- 17919721 TI - Placental IGF2 expression in normal and intrauterine growth restricted (IUGR) pregnancies. PMID- 17919720 TI - The use of keratin biomaterials derived from human hair for the promotion of rapid regeneration of peripheral nerves. AB - The management of trauma-associated nerve defects is difficult because of the absence of autologous donor motor or sensory nerves. Pre-clinical development and clinical experience has shown that damaged nerves can be surgically repaired using a tubular conduit interposed across the defect. Acceptable patient outcomes are achieved so long as the gap distance does not exceed a few centimeters. Although research in animals has demonstrated that nerve repair can be facilitated across slightly larger gaps by introducing a biomaterial filler into the conduit lumen, these biomaterials are not typically "neuroinductive" (i.e. capable of acting directly on regenerative cells to enhance nerve tissue formation beyond clinical limits). Moreover, their use does not often result in functional recovery equivalent to nerve autograft, the clinical gold standard. Here we show that a biomaterial gel made from the proteins found in human hair can mediate a robust nerve regeneration response, in part through activation of Schwann cells. In vitro, keratins extracted from human hair enhance the activity of Schwann cells by a chemotactic mechanism, increase their attachment and proliferation, and up-regulate expression of important genes. Moreover, these characteristics translate to improved functional nerve recovery in an animal model. These results suggest that a biomaterial derived from human hair keratins is neuroinductive and can facilitate an outcome comparable to autograft in a nerve injury model. PMID- 17919722 TI - ELISA and HPLC methods for atrazine and simazine determination in trophic chains samples. AB - The aim of the research was to determine optimal conditions for atrazine determination in trophic chain samples by means of an antigen-coated tube enzyme linked immunosorbent assay (ELISA). The ELISA method was used for analysis of a selection of samples and the results and method requirement compared with HPLC. The 2 h competitive ELISA showed a minimum detection limit of 0.05 ng mL(-1) and a dynamic range 0.1-2 ng mL(-1). Investigation of atrazine concentration in a selection of trophic chain samples indicated that the content of atrazine (microg kg(-1)) in soil samples was 3.2-85.4, vegetable roots 32.9-148.9, green parts of plants 67.7-136.4, cereals 42.4-91.5 and samples of animal origin 1.3-8.4. The correlation between results obtained by HPLC and ELISA methods was 0.97. In addition, simazine content was determined by the HPLC method in which the detection limits were 0.2 microg g(-1) for atrazine and 0.3 microg g(-1) for simazine. The content (microg kg(-1)) of simazine in soil samples was 13.5-15.5, in vegetables roots 29.5-93.7, in green parts of plants 34.6-72.6 and in cereals 158-189. The study demonstrates the utility and convenience of the simple, practical and cost-effective ELISA method in a non-immunoassay laboratory for the analysis of food and environmental samples. The method is ideal for the rapid screening of large numbers of samples in laboratories where access to HPLC facilities is limited or lacking. In addition the investigation demonstrates the presence of significant levels of atrazine and simazine in trophic chain samples collected from different areas of the region. As expected, the highest concentration of both herbicides was found in plants. PMID- 17919723 TI - Potential marker enzymes and metal-metal interactions in Helisoma duryi and Lymnaea natalensis exposed to cadmium. AB - In this study, we investigate the effects of exposure to cadmium and copper on Lymnaea natalensis and Helisoma duryi. The snails were dosed with Cd2+ or Cu2+ for a period of 96h. Snails dosed with Cd accumulated the metal significantly (P<0.05) in tissues but not in shells. Mortality was observed at approximately 1mg Cd/l of culture water. In tissues and shells of snails dosed with Cd or Cu, synergistic and antagonistic metal-metal interactions involving Cd, Cu, Zn, and Pb were observed and these may affect metal toxicity. Glutamate dehydrogenase, aspartate aminotransferase, and alanine aminotransferase were assayed in whole snail tissue sub-cellular fractions of Cd-dosed snails. Generally, enzyme activity significantly increased at lower concentrations of Cd but decreased at high concentrations of the metal. However, mitochondrial alanine aminotransferase activity progressively declined with increasing Cd concentration. The changes in some of the enzymes' activities suggest biomarker potential. PMID- 17919724 TI - Arsenic fractions and enzyme activities in arsenic-contaminated soils by groundwater irrigation in West Bengal. AB - A study for the effect of arsenic (As) contamination on beta-glucosidase, urease, acid-/alkaline-phosphatase, and arylsulphatase activities was conducted in As contaminated soils of West Bengal, India. The studied soils show low total As contents relative to those in other As-contaminated sites. A sequential extraction technique was used to quantify water soluble, exchangeable, carbonate bound, Fe/Mn oxide bound, organic bound, and residual As fractions. Arsenic concentrations in the two most labile fractions (i.e., water soluble and exchangeable fractions) were generally low. The studied enzymes were significantly and negatively correlated with water soluble and exchangeable As forms but did not show any significant correlations with other forms, indicating that water soluble and exchangeable forms exert a strong inhibitory effect on the soil enzyme activities. It is thus suggested that the enzyme activity can be helpful in assessing the effects of As on biochemical quality of soils. PMID- 17919726 TI - Central neurophysiology and dopaminergic control of ejaculation. AB - Although premature ejaculation (PE) represents the most common male sexual dysfunction, brain mechanisms controlling ejaculatory process remain poorly understood. Recently a group of neurons, identified in the lumbar spinal cord, has been proposed to constitute a spinal ejaculation generator. This key site in ejaculation control, relaying sensory inputs to the brain, is under supraspinal excitatory (medial preoptic area, paraventricular nucleus of the hypothalamus) and inhibitory (nucleus paragigantocellularis) controls. Activation of brain excitatory areas by dopamine (DA) or DA agonists being demonstrated to facilitate ejaculation, it seems particularly interesting to further understand the implication of central DA in the complex process leading to ejaculation. Moreover, the fact that dopaminergic pathways are involved in sexual behavior and that DA release in some brain regions is an important facilitator of male sexual behavior reinforces the crucial implication of DA. Clearly, a better understanding of DA incerto-hypothalamic pathways and targeting brain DA receptor subtypes mediating ejaculation (especially D(3) receptors) will benefit the development of new pharmacological strategies to treat ejaculatory dysfunction including PE. PMID- 17919727 TI - The relationship between alcoholic cerebellar degeneration and cognitive and emotional functioning. AB - Although it is now widely acknowledged that the cerebellum contributes to the modulation of higher-order cognitive and emotional functions, this relationship has not been extensively explored in perhaps the largest group of individuals with cerebellar damage, chronic alcoholics. Localised damage to the cerebellum has been associated with a specific constellation of deficits and has been termed the 'cerebellar cognitive affective syndrome' (CCAS) [Schmahmann, J.D., Sherman, J.C., 1998. The cerebellar cognitive affective syndrome. Brain 121, 561-579]. The CCAS describes a profile of impairments, including deficits in executive functioning and visuospatial skills, language disruption and altered personality and affective behaviour. It is conceivable that the CCAS may also develop in a subgroup of alcoholics with alcoholic cerebellar degeneration and may in part account for a proportion of the cognitive and affective deficits commonly observed with the condition. While evidence has emerged supporting such a relationship, methodological limitations and the lack of theoretically driven investigation of the contribution of cerebellar dysfunction to cognitive and emotional functioning in chronic alcoholics, preclude definitive conclusions being drawn. PMID- 17919729 TI - Enchytraeus albidus (Enchytraeidae): a test organism in a standardised avoidance test? Effects of different chemical substances. AB - Enchytraeids (Enchytraeus albidus) directly improve the pore structure of the soil and are indirectly involved in regulating the degradation of organic matter. Due to their behavior they are able to avoid unfavorable environmental conditions. Avoidance tests allow a first assessment of toxicity of a contaminated or spiked soil within 48 h, by using the reaction of the enchytraeids as measurement endpoint. In this period, the organisms can choose between the control soil and the test soil. In the tests reported here, enchytraeids were exposed to LUFA 2.2 soil spiked with the following set of toxic substances: copper chloride, zinc chloride, cadmium chloride, phenmedipham, benomyl, carbendazim, dimethoate, atrazine, pentachlorophenol, chlorpyriphos, lindane, TBTO, Linear Alkylbenzene Sulfonates (LAS) and boric acid. Different chemical concentrations were tested. EC50s ranged from 8 mg/kg (Carbendazim) to >1000 mg/kg (e.g. LAS). While the tested heavy metals showed clear dose-response relationships, the effect pattern differed considerably in the tests with organic chemicals, e.g. no avoidance behaviour was observed in LAS, even at very high doses. Here we proposed to standardize the Enchytraeid avoidance test in a way similar to what is currently done for the earthworm and collembolan avoidance tests by the International Standardisation Organisation (ISO). PMID- 17919725 TI - How do the basal ganglia contribute to categorization? Their roles in generalization, response selection, and learning via feedback. AB - This article examines how independent corticostriatal loops linking basal ganglia with cerebral cortex contribute to visual categorization. The first aspect of categorization discussed is the role of the visual corticostriatal loop, which connects the visual cortex and the body/tail of the caudate, in mapping visual stimuli to categories, including evaluating the degree to which this loop may generalize across individual category members. The second aspect of categorization discussed is the selection of appropriate actions or behaviors on the basis of category membership, and the role of the visual corticostriatal loop output and the motor corticostriatal loop, which connects motor planning areas with the putamen, in action selection. The third aspect of categorization discussed is how categories are learned with the aid of feedback linked dopaminergic projections to the basal ganglia. These projections underlie corticostriatal synaptic plasticity across the basal ganglia, and also serve as input to the executive and motivational corticostriatal loops that play a role in strategic use of feedback. PMID- 17919728 TI - The link between dyslexic traits, executive functioning, impulsivity and social self-esteem among an offender and non-offender sample. AB - PURPOSE: The current study had two core aims; first to explore the link between dyslexic traits and other aspects of functioning among a sample of offenders and non-offenders (students); and, second, to explore if dyslexic traits were over represented among offenders. A subsidiary aim was to explore if the results were influenced by an offender's current index offence (i.e. violent versus non violent). METHOD: Ninety-two adult male participants took part: sixty offenders and thirty-two non-offenders. All completed a structured interview assessing dyslexic traits, namely the Dyslexia Adult Screening Test battery (DAST: Fawcett and Nicholson, 1998). Participants also completed a measure of executive functioning (Benton Word Fluency Test, Benton, A. (1968) Differential behavioural effects in frontal lobe disease. Neuropsychologica, 6, 53-60), impulsivity (Barratt Impulsiveness Scale: BIS-II, Barratt, E.S. (1994), Impulsiveness and Aggression. In J. Monahan and H.J. Steadman (Eds.), Violence and Mental Disorder: Developments in Risk Assessment (pp.61-79). Chicago: University of Chicago Press.) and social self-esteem (Texas Social Behavior Inventory: TSBI, Helmreich and Stapp, 1974). RESULTS: Offenders presented with more dyslexic traits than non offenders, with those with violent index offences presenting with more traits than those with non-violent index offences. Offenders performed poorly on assessments of executive functioning when compared with non-offenders. Dyslexic traits were predicted most significantly by executive functioning difficulties followed by decreased social self-esteem. There was a trend for increased impulsivity to correlate with increased dyslexic traits. Dyslexic traits were also predictive of membership to the offender group whereas impulsivity, executive functioning or social self-esteem was not. CONCLUSION: Preliminary evidence is provided for increased dyslexic traits among offenders compared to non-offenders. The study highlights the correlates of dyslexic traits. The implications of these findings for future research are outlined. PMID- 17919730 TI - Constructed wetlands for environmental pollution control: a review of developments, research and practice in Ireland. AB - For the purpose of synthesizing a compendium of efforts aimed at environmental pollution control through the use of constructed wetlands systems (CWs) in Ireland, a detailed review of CWs was undertaken. Emphasis was placed on the diverse range of development, practice and researches on CWs technology, placing them in the overall context of the need for low-cost and sustainable wastewater treatment systems. The potential use of CWs in protecting estuarine quality within the current legislative framework is considered, as well as the emerging concept of integrated constructed wetlands (ICWs). In addition, an assessment of the efficiency of CWs in operation in Ireland towards abating environmental pollution was done, and compared with CWs operating in other European countries. The need for sufficient and appropriate data to assist in further development of CWs and modelling studies, and instilling confidence in the public is also highlighted. PMID- 17919731 TI - Hyphal growth patterns and recurrence of fungal keratitis after lamellar keratoplasty. AB - PURPOSE: To evaluate growth patterns of fungal pathogens in corneas and recurrence of fungal keratitis after lamellar keratoplasty (LK). DESIGN: Retrospective noncomparative study. PARTICIPANTS: One hundred seventy-four patients (174 eyes) with fungal keratitis who underwent LK at Shandong Eye Institute from January 2000 through November 2006. METHODS: Medical records of each patient were retrospectively reviewed. Hyphal growth patterns in corneas were evaluated by histopathological examination. Fungal recurrence after LK was observed during the follow-up. MAIN OUTCOME MEASURES: Pathogens, hyphal growth patterns, and postoperative fungal recurrence. RESULTS: The pathogens were Fusarium (85.1%), Aspergillus (6.3%), Alternaria (4.6%), Penicillium (2.3%), and Candida (1.7%). Most Fusarium hyphae (91.2%) lay parallel to the corneal stromal lamellae, whereas most Aspergillus (90.9%) grew vertically. Recurrence of fungal keratitis was found in 15 patients (8.6%) after LK, and the pathogens were F. oxysporum (33.3%), F. solani (26.7%), F. moniliforme (13.3%), Aspergillus flavus (13.3%), Aspergillus fumigatus (6.7%), and Aspergillus terreus (6.7%). In cases of fungal recurrence, the majority of hyphae (80%) grew vertically. There was a higher recurrence rate in patients with vertically growing hyphae (46.2%) than in those with horizontally growing hyphae (2%) (chi(2) = 54.664, P<0.001), as well as in those with Aspergillus keratitis (36.4%) versus those with Fusarium keratitis (7.4%) (chi(2) = 10.031, P = 0.002). Reproducibility of the fungal recurrence rate was moderate in the patients with different hyphal growth patterns (kappa = 0.534) but poor in those with different fungal pathogens (kappa = -0.044). CONCLUSIONS: Hyphal growth patterns in corneas differ not only in the same fungal genus but also in the same species. The fungal recurrence rate after LK in patients with hyphae growing horizontally is much lower than that in those with hyphae growing vertically. Growth patterns of fungal pathogens may be an important factor for fungal recurrence after LK. PMID- 17919734 TI - Information transfer rate in fMRI experiments measured using mutual information theory. AB - Information theory provides a mathematical framework for analysis of fMRI experiments. By modeling the fMRI experiment as a communication system, various results from information theory can be applied to measure information transfer rate in fMRI experiments. The information transfer rate has important implications for design and analysis of brain-computer interface (BCI) experiments. A key factor in the effective implementation of BCI techniques is to achieve maximum information transfer rate. In this report, mutual information rate (MIR) was used to evaluate the efficiency of alternative experimental designs. The channel capacity, a fundamental physical limit on the rate at which information can be extracted from an fMRI experiment, was estimated and compared with the theoretical limit specified by the Hartley-Shannon Theorem. We present an information theory framework for the analysis of fMRI time-series assuming a known hemodynamic response function. Using MIR to evaluate fMRI experimental designs, we show that block lengths of 3-5s have maximum information transfer rates. For designs with shorter block lengths, the MIR is limited by the channel capacity. For experimental designs with longer block lengths, the MIR is limited by the low source information transmission rate. PMID- 17919733 TI - Some historical reflections on the neural control of locomotion. AB - Thought on the neural control of locomotion dates back to antiquity. In this article, however, the focus is more recent by starting with some major 17th century concepts, which were developed by Rene Descartes, a French philosopher; Thomas Willis, an English anatomist; and Giovanni Borelli, an Italian physiologist and physicist. Each relied on his personal expertise to theorize on the organization and control of movements. The 18th and early 19th centuries saw work on both the central and peripheral control of movement: the former most notably by Johann Unzer, Marie Jean-Pierre Flourens and Julien-Jean-Cesar Legallois, and the latter by Unzer, Jiri Prochaska and many others. Next in the 19th century, neurologists used human locomotion as a precise tool for characterizing motor pathologies: e.g., Guillaume Duchenne de Boulogne's description of locomotor ataxia. Jean-Martin Charcot considered motor control to be organized at two levels of the central nervous system: the cerebral cortex and the spinal cord. Maurice Philippson's defined the dog's step cycle and considered that locomotion used both central and reflex mechanisms. Charles Sherrington explained that locomotor control was usually thought to consist of a succession of peripheral reflexes (e.g., the stepping reflexes). Thomas Graham Brown's then contemporary evidence for the spinal origin of locomotor rhythmicity languished in obscurity until the early 1960s. By then the stage was set for an international assault on the neural control of locomotion, which featured research conducted on both invertebrate and vertebrate animal models. These contributions have progressively became more integrated and interactive, with current work emphasizing that locomotor control involves a seamless integration between central locomotor networks and peripheral feedback. PMID- 17919732 TI - Regulation of extracellular matrix remodeling and cell fate determination by matrix metalloproteinase stromelysin-3 during thyroid hormone-dependent post embryonic development. AB - Interactions between cells and extracellular matrix (ECM), in particular the basement membrane (BM), are fundamentally important for the regulation of a wide variety of physiological and pathological processes. Matrix metalloproteinases (MMP) play critical roles in ECM remodeling and/or regulation of cell-ECM interactions because of their ability to cleave protein components of the ECM. Of particular interest among MMP is stromelysin-3 (ST3), which was first isolated from a human breast cancer and also shown to be correlated with apoptosis during development and invasion of tumor cells in mammals. We have been using intestinal remodeling during thyroid hormone (TH)-dependent amphibian metamorphosis as a model to study the role of ST3 during post-embryonic tissue remodeling and organ development in vertebrates. This process involves complete degeneration of the tadpole or larval epithelium through apoptosis and de novo development of the adult epithelium. Here, we will first summarize expression studies by us and others showing a tight spatial and temporal correlation of the expression of ST3 mRNA and protein with larval cell death and adult tissue development. We will then review in vitro and in vivo data supporting a critical role of ST3 in TH induced larval epithelial cell death and ECM remodeling. We will further discuss the potential mechanisms of ST3 function during metamorphosis and its broader implications. PMID- 17919735 TI - Evaluation of corticospinal axon loss by fluorescent dye tracing in mice with experimental autoimmune encephalomyelitis. AB - In both multiple sclerosis (MS) patients and experimental autoimmune encephalomyelitis (EAE) animals, axon loss has been demonstrated to correlate with neurological disability. However, it is difficult to accurately determine the location and severity of axonal damage since the lesion in MS or EAE is disseminated and is frequently in a relapsing-remitting mode. The corticospinal system is the only direct pathway from the motorsensory cortex to the spinal cord, and the major neural pathway for control of voluntary movement. Moreover, it is frequently involved in the pathological process of the disease. To evaluate corticospinal tract (CST) axon loss in EAE mice, we developed a direct tracing method with a fluorescent neuronal tracer DiI which was injected into the primary motor cortex and sensorimotor cortex to label the pyramidal neurons. The lesion location in the spinal cord and axon disruption were indicated by dye leakage. Using the EAE induced axon reduction as an index of the extent of axonal damage, our data showed a high correlation between the axonal loss and the behavioral outcome score in the EAE mice. The results were consistent with the axonal Bielschowsky silver staining. Thus, this CST tracing method permits monitoring of the axonal damage in EAE. PMID- 17919736 TI - Remitted depression moderates self-injurious behavior in personality-disordered research volunteers. AB - The moderating effects of depression on self-injurious behavior among personality disordered individuals (N=40) were examined. Self-injurious behavior (SIB) was assessed using a well-validated laboratory measure. Remitted depression was associated with greater sensitivity to self-aggressive cues, indicating that remitted depression may be a risk factor for SIB. PMID- 17919737 TI - Relationship between cardiovagal modulation and psychotic state in patients with paranoid schizophrenia. AB - Disturbed autonomic nervous system (ANS) function in schizophrenia might contribute to increased cardiovascular mortality. We obtained heart rate variability indices from 40 unmedicated schizophrenic patients and 58 matched controls. Mainly we found that patients displaying stronger psychotic symptoms as assessed by the Brief Psychiatric Rating Scale exhibit more severe cardiac ANS disturbances compared with controls. PMID- 17919740 TI - Antibody-secreting cells in the central nervous system in an animal model of MS: Phenotype, association with disability, and in vitro production of antibody. AB - Little is known about antibody production within the central nervous system, called intrathecal antibody production (ITAbP). Mice infected with Theiler's murine encephalomyocarditis virus (TMEV) develop an immune-mediated demyelinating disease(TMEV-IDD), characterized by progressive weakness associated with robust ITAbP, CNS inflammation and demyelination. TMEV-IDD represents an excellent model of human multiple sclerosis (MS), especially its progressive disability. The mechanism of the weakness in this disease is unknown but there is considerable evidence that it is immune-mediated. Our hypothesis was that the disability in the model is induced by robust ITAbP by antibody-secreting cells(ASCs) in the CNS. We further hypothesized that these ASCs are predominantly CD138+ plasma cells, driven by the persistence of virus at high levels in the central nervous system (CNS) as well as high levels of B-cell activating factor (BAFF). In order to test these hypotheses we infected mice with TMEV, and correlated measures of ITAbP with disability. Measures of ITAbP were ELISpot and IgG gene expression, while disability was tested by Rotarod. We found that disability was highly correlated with ITAbP, assayed by number of CNS ASCs as well as amount of IgG mRNA. CNS ASCs were primarily CD138+, and produced high levels of IgG enriched for reactivity to TMEV. There were high levels of BAFF production in the CNS, but these levels were only minimally higher in infected mice than in uninfected controls. TMEV and IgG RNA were distributed throughout the spinal cord. These data are the first demonstrating that ITAbP is highly correlated with disability in TMEV infection, an excellent model of human MS. Our data raise the possibility that ITAbP contributes significantly to disability, both in this model and in MS. PMID- 17919739 TI - Dissociation of spinal microglia morphological activation and peripheral inflammation in inflammatory pain models. AB - We compared the effects of peripheral Freund's Complete Adjuvant (CFA) and formalin injection on spinal microglia activation. Both qualitative and quantitative analyses showed signs of microglia activation on the ipsilateral side of the lumbar dorsal horn on day 3, day 7 and day 14 after formalin injection. However, significant microglia morphological alteration was not found in the CFA model. At the injection site in the paw, CFA injection induced considerably more inflammation than formalin injection. Although spinal microglia might be activated in inflammatory pain models, morphologically, spinal microglia activation was not closely correlated with peripheral inflammation. PMID- 17919738 TI - G protein-coupled receptors and the modification of FcepsilonRI-mediated mast cell activation. AB - By releasing multiple pro-inflammatory mediators upon activation, mast cells are critical effector cells in the pathogenesis of allergic inflammation. The traditional viewpoint of antigen-dependent mast cell activation is that of a Th(2)-driven process whereby antigen-specific IgE molecules are produced by B cells followed by binding of the IgE to high affinity IgE receptors (FcepsilonRI) expressed on mast cells. Subsequent antigen-dependent aggregation of the FcepsilonRI initiates an intracellular signalling cascade that culminates in mediator release. Mast cell responses, including cell growth, survival, chemotaxis, and cell adhesion, however, can also be regulated by other receptors expressed on mast cells. Furthermore, FcepsilonRI-mediated mast cell mediator release can be significantly modified by ligation of specific classes of these receptors. One such class of receptors is the G protein-coupled receptors (GPCR). In this review, we describe how sub-populations of GPCRs can either enhance or inhibit FcepsilonRI-mediated mast cell activation depending on the particular G protein utilized for relaying signalling. Furthermore, we discuss the potential mechanisms whereby the signalling responses utilized by the FcepsilonRI for mast cell activation are influenced by those initiated by GPCRs to produce these diverse responses. PMID- 17919741 TI - An investigation of the significance of Actinomycosis in tonsil disease. AB - OBJECTIVES: The objective of the study was to establish the incidence of Actinomycosis in the tonsils of children undergoing tonsillectomy or adenotonsillectomy, and to evaluate its role in clinical tonsillar disease. METHODS: This was a prospective controlled study done at the Red Cross Children's Hospital in Cape Town, South Africa over an 8-month period and included all children undergoing tonsillectomy or adenotonsillectomy. All resected tonsils were examined for the presence of Actinomycosis and any signs of significant cryptitis or active tonsillitis. A comparison was made in the incidence of Actinomycosis in children with obstructive sleep apnoea, recurrent tonsillitis or obstructive sleep apnoea and recurrent tonsillitis. The data was further analysed to determine the statistical significance of the association between Actinomycosis of the tonsils and age, sex and histopathological and clinical diagnosis. RESULTS: A total of 344 tonsils were analysed on 172 patients. We found 20 patients (11.6%) with Actinomycosis in the tonsils. The mean age of patients with Actinomycosis was 7.25 years and without Actinomycosis was 5.4 years (p=0.002). Most specimens (16) had no evidence of tissue reaction to Actinomyces, and their presence was found to be due to colonisation of the tonsils only. Actinomycosis was present in 11% of patients with obstructive sleep apnoea, 11% of patients with recurrent tonsillitis and in 9% with obstructive sleep apnoea and recurrent tonsillitis. The difference in incidence of Actinomycosis between these three groups (p=0.94), and between the recurrent tonsillitis group alone compared to the obstructive group (p=0.83), was not statistically significant. There was therefore no statistical significance found between Actinomyces and OSA+/- recurrent tonsillitis. CONCLUSIONS: There was no correlation found between the presence of tonsillar Actinomycosis and recurrent tonsillitis and/or obstructive tonsillar hypertrophy. Histopathologic findings showed no evidence of tissue reaction to Actinomyces and its presence was found to be due to colonisation of the tonsils only. The series did however show a statistically significant correlation between Actinomycosis colonisation and age with Actinomycosis being more common in older children, especially those over 5 years of age. PMID- 17919742 TI - An indirect immunocolorimetric assay to detect rubella virus infected cells. AB - An indirect immunocolorimetric assay (ICA) to detect rubella virus infected cells by the naked eye was developed. This assay was as sensitive and specific as an indirect immunofluorescent assay (IFA), could detect as little as 10 plaque forming units (pfu) of rubella virus in the initial inoculum and could detect viruses from throat swabs. This assay detected infection with viruses of all nine rubella virus genotypes available for testing. It could be utilized for virus quantitation and a neutralization assay. In addition to detecting rubella virus infection, this assay also allowed us to confirm measles virus infection. This assay should be useful for virus isolation from clinical samples. PMID- 17919743 TI - Activity of the neuraminidase inhibitor A-315675 against oseltamivir-resistant influenza neuraminidases of N1 and N2 subtypes. AB - Clinical use of the neuraminidase inhibitor (NAI) oseltamivir has been associated with the emergence of viral resistance resulting from subtype-specific neuraminidase (NA) mutations. In this study, we evaluated the impact of the most frequent oseltamivir-resistant NA mutations including E119V, H274Y, R292K and N294S on the susceptibility profile to a novel NAI (A-315675) using recombinant NA proteins of N1 and N2 subtypes and also selected oseltamivir-resistant influenza H1N1 and H3N2 viruses. In the N1 subtype, recombinant NA proteins containing mutations H274Y and N294S previously associated with resistance to oseltamivir (754- and 197-fold increases in IC(50) values, respectively, compared to WT) remained susceptible to A-315675 (2.5- and 2-fold increases in IC(50) values(,) respectively). In the N2 subtype, NA proteins harboring mutations E119V and R292K conferring high levels of resistance to oseltamivir (1016- and >10,000 fold increases in IC(50) values, respectively) had IC(50) values that increased by only 1.5- and 13-fold, respectively, against A-315675. Similar susceptibility patterns to A-315675 were obtained when testing recombinant H1N1 mutant viruses (H274Y and N294S) and clinical H3N2 mutants (E119V). The V116A and I117V mutations, previously associated with oseltamivir resistance in H5N1 viruses, were susceptible to oseltamivir when tested in the H1N1 background suggesting a strain-specific impact of these mutations. These results confirm the potent inhibitory effect of A-315675 against oseltamivir-resistant influenza viruses of the N1 and N2 subtypes and support the clinical development of its bioavailable prodrug A-322278. PMID- 17919745 TI - Phenotypic segregation of aphakia and Pitx3-null mutants reveals that Pitx3 deficiency increases consolidation of specific movement components. AB - Deficiency of the meso-diencephalic dopamine (mdDA) neuron specific transcription factor Pitx3 in aphakia (ak) mice results in the loss of the substantia nigra compacta (SNc). Concomitantly, reduced spontaneous locomotor behavior, symptoms reminiscent to those in Parkinson's disease, has been reported. However, the ak mouse line originates from the 1960s and has been compared to C57BL/6J inbred controls. Therefore, to define Pitx3 gene function in baseline and novelty induced locomotor behavior and mdDA neuronal activity, we analyzed Pitx3 deficiency in a controlled genetic and epigenetic background. The analysis implicated that, in contrast to the controversial and previously reported hypo activity in ak mice, Pitx3-/- mice showed normal dark phase motor activity levels. Our data also revealed that ak and Pitx3-/- mice both display a similar neuro-anatomical and physiological phenotype, and, interestingly, showed increased spontaneous home cage activity levels during their habitual sleep phase. Further behavioral analysis revealed that both ak and Pitx3-/- mice have reduced transitions but increased consolidation of specific locomotor behaviors, such as rearing and horizontal movement. Thus, Pitx3 is not involved in the expression of nighttime motor activity levels, but is critical for selective mdDA neuronal activity and associated with increased consolidation of movement. PMID- 17919744 TI - Infusions of 3alpha,5alpha-THP to the VTA enhance exploratory, anti-anxiety, social, and sexual behavior and increase levels of 3alpha,5alpha-THP in midbrain, hippocampus, diencephalon, and cortex of female rats. AB - 17beta-Estradiol (E2) and progesterone (P4) influence the onset and duration of sexual behavior and are also associated with changes in behaviors that may contribute to mating, such as exploration, anxiety, and social behaviors (socio sexual behaviors). In the midbrain ventral tegmental area (VTA), the P4 metabolite, 5alpha-pregnan-3alpha-ol-20-one (3alpha,5alpha-THP), modulates lordosis of E2-primed rodents; 3alpha,5alpha-THP can also influence anxiety and social behaviors. To examine if 3alpha,5alpha-THP in the VTA mediates socio sexual behaviors, we infused 3alpha,5alpha-THP to the VTA of diestrous and proestrous rats. As expected, proestrous, compared to diestrous, rats showed more exploratory (open field), anxiolytic (elevated plus maze), pro-social (partner preference, social interaction), and sexual (paced mating) behavior and had increased E2, P4, dihydroprogesterone (DHP), and 3alpha,5alpha-THP in serum, midbrain, hippocampus, diencephalon, and cortex. Infusions of 3alpha,5alpha-THP to the VTA, but not control sites, such as the substantia nigra (SN) or central grey (CG), of diestrous rats produced behavioral and endocrine effects akin to that of proestrous rats and increased DHP and 3alpha,5alpha-THP levels in midbrain, hippocampus, and diencephalon. Levels of DHP and 3alpha,5alpha-THP, but neither E2 nor P4 concentrations, in midbrain, hippocampus, diencephalon, and/or cortex were positively correlated with socio-sexual behaviors. Thus, 3alpha,5alpha-THP infusions to the VTA, but not SN or CG, can enhance socio sexual behaviors and increase levels in midbrain, hippocampus, and diencephalon. PMID- 17919746 TI - Vasoactive intestinal peptide rescues cultured rat myenteric neurons from lipopolysaccharide induced cell death. AB - The role of the enteric nervous system in intestinal inflammation is not fully understood and the plethora of cellular activities concurrently ongoing in vivo renders intelligible studies difficult. In order to explore possible effects of bacterial lipopolysaccharide (LPS) on enteric neurons we utilised cultured myenteric neurons from rat small intestine. Exposure to LPS caused markedly reduced neuronal survival and increased neuronal expression of vasoactive intestinal peptide (VIP), while the expression of Toll-like receptor 4 (TLR4) was unchanged. TLR4 was expressed in approximately 35% of all myenteric neurons irrespective of if they were cultured in the presence or absence of LPS. In neurons cultured in medium, without LPS, 50% of all TLR4-immunoreactive neurons contained also VIP. Addition of LPS to the neuronal cultures markedly increased the proportion of TLR4-immunoreactive neurons also expressing VIP, while the proportion of TLR4 neurons devoid of VIP decreased. Simultaneous addition of LPS and VIP to the neuronal cultures resulted in a neuronal survival comparable to controls. CONCLUSIONS: LPS recognition by myenteric neurons is mediated via TLR4 and causes neuronal cell death. Presence of VIP rescues the neurons from LPS induced neurodegeneration. PMID- 17919747 TI - Protective role of adrenomedullin in burn-induced remote organ damage in the rat. AB - Clinical and experimental research findings suggest that a local burn insult produces oxidant-induced organ changes as evidenced by increased lipid peroxidation in lung, liver and gut. Adrenomedullin (AM), a potent vasodilator, was originally isolated from pheochromocytoma cells, and has been identified in other tissues. In this study, we investigated the potential role of AM in burn induced remote organ damage in rats. Sprague-Dawley rats (250-300 g) were treated with either AM (100 ng/kg, subcutaneously) or saline 10 min before burn insult which covers 30% of total body surface area and were decapitated 24 h after the burn insult. Trunk blood was collected and analyzed for liver and kidney functions and for determination of TNF-alpha levels. The liver, lung and kidney samples were taken for histologic evaluation and for measurement of malondialdehyde (MDA) level, myeloperoxidase (MPO) activity and chemiluminescence levels. The data revealed that AM treatment resulted in a significant protection in tissues tested against burn injury via suppression of lipid peroxidation, tissue neutrophil infiltration, oxidant generation and via decreasing circulating levels of the pro-inflammatory cytokine TNF-alpha. AM treatment was also effective in attenuating hepatic and kidney dysfunction due to burn injury, suggesting that peripherally AM administration may protect the tissues against burn-induced injury. PMID- 17919748 TI - Heterogeneous nuclear ribonucleoprotein A3 binds single-stranded telomeric DNA and inhibits telomerase extension in vitro. AB - Telomeres are dynamic DNA-protein complexes at the end of linear chromosomes. Maintenance of functional telomeres is required for chromosome stability, and to avoid the activation of DNA damage response pathway and cell cycle arrest. Telomere-binding proteins play crucial roles in the maintenance of functional telomeres. In this study, we employed affinity pull-down and proteomic approach to search for novel proteins that interact with the single-stranded telomeric DNA. The proteins identified by two-dimensional gel electrophoresis were further characterized by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and MALDI-TOF-TOF tandem MS. Among the five identified proteins, we report here the biochemical properties of a novel protein, hnRNP A3. The purified hnRNP A3 bound specifically to G-rich strand, but not to C-rich strand or double-stranded telomeric DNA. The RRM1 (RNA recognition motif 1) domain, but not RRM2, of hnRNP A3 is sufficient to confer specific binding to the telomeric sequence. In addition, we present evidence that hnRNP A3 can inhibit telomerase extension in vitro. These biochemical properties of hnRNP A3 suggest that hnRNP A3 can participate in telomere regulation in vivo. PMID- 17919750 TI - Sterile abscess formation as a complication of hydrophilic radial artery cannulation. AB - A sterile inflammatory response has been observed at the access site in a small number of patients who have undergone radial artery cannulation with a hydrophilic sheath. This reaction generally appears from several days to weeks after use of the sheath, and while a distinct cause has not been established it is not thought to be due to infection. This particular risk of use of hydrophilic radial sheaths must therefore be weighed against its potential benefits. PMID- 17919749 TI - Disruption of aldo-keto reductase genes leads to elevated markers of oxidative stress and inositol auxotrophy in Saccharomyces cerevisiae. AB - A large family of aldo-keto reductases with similar kinetic and structural properties but unknown physiological roles is expressed in the yeast Saccharomyces cerevisiae. Strains with one or two AKR genes disrupted have apparently normal phenotypes, but disruption of at least three AKR genes results in a heat shock phenotype and slow growth in inositol-deficient culture medium (Ino(-)). The present study was carried out to identify metabolic or signaling defects that may underlie phenotypes that emerge in AKR deficient strains. Here we demonstrate that pretreatment of a pentuple AKR null mutant with the anti oxidative agent N-acetyl-cysteine rescues the heat shock phenotype. This indicates that AKR gene disruption may be associated with defects in oxidative stress response. We observed additional markers of oxidative stress in AKR deficient strains, including reduced glutathione levels, constitutive nuclear localization of the oxidation-sensitive transcription factor Yap1 and upregulation of a set of Yap1 target genes whose function as a group is primarily involved in response to oxidative stress and redox balance. Genetic analysis of the Ino(-) phenotype of the null mutants showed that defects in transcriptional regulation of the INO1, which encodes for inositol-1-phosphate synthase, can be rescued through ectopic expression of a functional INO1. Taken together, these results suggest potential roles for AKRs in oxidative defense and transcriptional regulation. PMID- 17919752 TI - Unusual left atrial appendage thrombus in a neonate with normal heart after sustained supraventricular tachycardia. AB - Left atrial appendage thrombi in neonates are uncommon. We describe a neonate with a large thrombus in the left atrial appendage detected by echocardiography after a paroxystic sustained supraventricular tachycardia associated with intermittent Wolff-Parkinson-White syndrome. Mechanisms and therapy of atrial thrombosis in neonates are briefly discussed. PMID- 17919751 TI - Coronary stent implantation may seal the inflammatory response in patients with acute coronary syndromes. AB - The change in CD40L levels before and 24 h coronary stenting was compared in patients presenting with acute coronary syndromes and for elective angioplasty. Baseline CD40L levels were lower in stable subjects, whereas unstable patients had a significant decrease of CD40L after stenting. This might suggest that coronary stenting, despite of being a local therapy, can help to seal a systemic inflammatory response in acute coronary syndromes. PMID- 17919753 TI - Citalopram induced torsade de pointes, a rare life threatening side effect. AB - Acquired Long QT syndrome is a disorder caused by medications, electrolyte imbalances, and drug interactions. This syndrome is associated with an increased risk of a characteristic life-threatening cardiac arrhythmia, known as torsade de pointes (TdP). In the setting of Long QT syndrome (LQTS), selective serotonin reuptake inhibitors (SSRIs) can precipitate TdP. We report the first case of LQTS and TdP induced by citalopram in the United States. After discontinuation of citalopram, the QT/QTc interval normalized after 3 days and resolved further episodes of TdP. Patients on citalopram should be monitored closely for QT/QTc interval to prevent torsade de pointes. PMID- 17919754 TI - Near miss sudden cardiac death on a young patient with repaired atrioventricular septal defect. AB - Patients with congenital heart disease often face the prospect of long-term haemodynamic or arrhythmic complications for which lifelong follow-up in specialist adult congenital heart disease (ACHD) centres is required. We describe the case of a 25-year-old man with repaired atrioventricular septal defect who was referred to our centre after a ventricular fibrillation arrest. Serial echocardiograms in previous years had shown progressive severe left ventricular outflow obstruction, but the patient had not been operated on as he was deemed asymptomatic and reluctant to consider surgery. Management and criteria for further intervention in ACHD patients often differ from those of patients with acquired heart disease and reliance on symptoms alone is not good practice and may prove catastrophic. PMID- 17919755 TI - Release pattern of cardiac troponin in left ventricular apical ballooning syndrome: insights into the mechanisms of stress cardiomyopathy. PMID- 17919758 TI - Genus-specific profile of acetic acid bacteria by 16S rDNA PCR-DGGE. AB - An effective method for grouping acetic acid bacteria (AAB) genera was defined and evaluated as a tool for preliminary screening of the major AAB species involved in vinegar production. Acetobacter, Gluconobacter, Gluconacetobacter, Asaia, Neoasaia, Saccharibacter, Frateuria and Kozakia AAB strains were screened on the basis of the 16S rDNA sequences using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique. The DGGE profile of all the strains tested, consisted of one single band of approximately 330 bp for each strain and allowed their clustering. The results obtained clearly reflected in silico phylogenetic analysis of the AAB species used in this study, in fact, the species with a higher 16S rDNA sequence homology showed a similar electrophoretic profile. In particular almost all the species belonging to the genus Gluconacetobacter showed a DGGE pattern nearly identical and well distinct from all the other AAB genera. Furthermore by PCR-DGGE it was possible to clearly group the species more frequently recovered from vinegar fermentation which are mainly distributed in the genera Acetobacter, Gluconobacter and Gluconacetobacter. PMID- 17919756 TI - Expression and functional characterization of gfpmut3.1 and its unstable variants in Staphylococcus epidermidis. AB - Reporter gene systems are an invaluable tool for investigation of gene transcription activity in eukaryotes and prokaryotes. In order to analyze the temporal and spatial resolution of gene expression patterns in situ and for quantitatively investigating gene expression, the green fluorescent protein (GFP) appears to be especially useful. GFP has been broadly used in various bacterial species, however, there is only limited knowledge about key biological properties in S. epidermidis. Here, the crucial influence of different ribosomal binding sites (RBS) on gfpmut3.1 translation initiation in S. epidermidis 1457 is demonstrated. Only by using the RBS of the delta-hemolysin promoter, after 24 hours a strong fluorescence signal was obtained. The half-life of GFPmut3.1 in S. epidermidis 1457 was significantly shorter than in E. coli (7 h vs. 24 h). GFPmut3.1 derivatives with shorter half-lives (GFP(AAV) and GFP(ASV)) did not reach sufficient quantitative protein levels, and the resulting low fluorescence limits their use as reporter genes in S. epidermidis. This work provides fundamental insights into gfpmut3.1 expression in S. epidermidis and describes the crucial determinants of its biological behavior in this species. In general, this study underlines the need to accurately characterize key biological properties of this transcription marker in gram-positive hosts. PMID- 17919759 TI - Effective approaches for the production of heterologous proteins using the Thermococcus kodakaraensis-based translation system. AB - We have previously established a system for cell-free protein synthesis that can be operated at high temperatures using the cell lysate of a hyperthermophilic archaeon, Thermococcus kodakaraensis. To apply this system to practical use in the field of heterologous protein production, the performance of our system in the synthesis of green fluorescent protein (GFP) was examined. As the wild-type GFP is a thermolabile protein, a thermostable GFP derivative (tGFP) was selected as a candidate for protein synthesis. The first attempt of tGFP synthesis at 60 degrees C using the system resulted in a detection of small amount of protein (<0.1 microg/mL) by Western blot analysis. Using a newly synthesized tGFP gene in which codon usage was optimized for T. kodakaraensis as a template, tGFP was clearly detectable at temperatures between 50 and 65 degrees C. The tGFP production was further enhanced over 10 microg/mL with the addition of stem-loop structure at the 3'-end of mRNA. Determination of fluorescences of tGFP in the reaction mixtures indicated that active tGFP constituted ca. 30% of the total protein synthesized. Addition of T. kodakaraensis chaperonin to the system significantly increased the ratio of active tGFP content to ca. 50%. Through these approaches to the system, the production of tGFP increased over 100-fold, and the yield of active tGFP synthesized reached to 6.5 microg/mL. PMID- 17919760 TI - Metabolic flux screening of Saccharomyces cerevisiae single knockout strains on glucose and galactose supports elucidation of gene function. AB - New methods for an extended physiological characterization of yeast at a microtiter plate scale were applied to 27 deletion mutants of Saccharomyces cerevisiae cultivated on glucose and galactose as sole carbon sources. In this way, specific growth rates, specific rates of glucose consumption and ethanol production were determined. Flux distribution, particularly concerning branching into the pentose phosphate pathway was determined using a new (13)C-labelling method using MALDI-ToF-mass spectrometry. On glucose, the growth was predominantly fermentative whereas on galactose respiration was more active with correspondingly lower ethanol production. Some deletion strains showed unexpected behavior providing very informative data about the function of the corresponding gene. Deletion of malic enzyme gene, MAE1, did not show any significant phenotype when grown on glucose but a drastically increased branching from glucose 6 phosphate into the pentose phosphate pathway when grown on galactose. This allows the conclusion that MAE1 is important for the supply of NADPH during aerobic growth on galactose. PMID- 17919761 TI - Post-translational modifications of Abutilon mosaic virus movement protein (BC1) in fission yeast. AB - The movement protein (MP) of Abutilon mosaic virus (AbMV, Geminiviridae) exhibited a complex band pattern upon gel electrophoresis indicating its post translational modification when expressed in AbMV-infected plants or, ectopically, in fission yeasts. High-resolution separation according to charge and molecular weight in acetic acid/urea polyacrylamide or sodium dodecyl sulphate polyacrylamide gels followed by western blot analysis revealed a pattern of AbMV MP from infected plants more related to that from fission yeast than from bacteria. For this reason, expression in fission yeast was established as an experimental system to study post-translational modifications of AbMV MP. Metabolic labelling with 32P-orthophosphate confirmed a phosphorylation of all MP variants suggesting multiple phosphorylation sites. Treatment with calf intestinal alkaline phosphatase removed this label completely, but was unable to eliminate all protein bands with lower electrophoretic mobility. Thus, multiple phosphorylations contribute to the complex migration behaviour of MP, but additional post-translational modifications may occur as well. PMID- 17919762 TI - Six-month multicentric, open-label, randomized trial of twice-daily injections of biphasic insulin aspart 30 versus multiple daily injections of insulin aspart in Japanese type 2 diabetic patients (JDDM 11). AB - To evaluate glycemic control using convenience-oriented biphasic insulin analog compared with intensified insulin therapy, we conducted a 6-month multicentric, open-label, randomized trial in Japanese insulin-naive patients with type 2 diabetes mellitus. A total of 160 adult patients at 19 centers were randomized into two groups: those who received twice-daily injections of biphasic insulin aspart 30 and those on three-times-daily injections of insulin aspart with or without NPH insulin (multiple daily injections). At 6 months, mean HbA(1c) decreased by approximately 2.5% in both groups. Reduction of HbA(1c) on both regimens was better in patients whose prior therapy before starting the study was only diet and exercise (-5.0%) than in patients who were previously taking oral antidiabetic agents (-1.0%). No incidence of major hypoglycemia was observed in either regimen. These results suggest that convenience-oriented insulin therapy using biphasic insulin analog is as useful as intensified insulin therapy with insulin analog for the treatment of type 2 diabetes mellitus over 6 months. Furthermore, early induction of insulin therapy in individuals hitherto using only diet and exercise may provide good glycemic control. This study suggests that convenience-oriented biphasic insulin aspart 30 might be a useful option for the treatment of type 2 diabetes mellitus, especially for insulin-naive patients over 6 months, although it should be changed to another regimen when expected efficacy is not obtained. PMID- 17919763 TI - Efficacy of insulin glargine and glimepiride in controlling blood glucose of ethnic Japanese patients with type 2 diabetes mellitus. AB - BACKGROUND: To evaluate the efficacy and safety of glimepiride plus insulin glargine in ethnic Japanese patients with type 2 diabetes mellitus (T2DM). METHODS: This 24-week, open-label, single-arm study was conducted in eight centers in Brazil. One hundred ethnic Japanese T2DM patients with inadequate glycemic control [HbA(1c): 8.0-11.0% and fasting plasma glucose (FPG)>or=140 mg/dL] on oral antidiabetic drugs (OADs) were enrolled. Patients were treated once daily with glimepiride 3mg (morning) and glargine (bedtime) with dose titration to achieve FPG 72-100mg/dL. RESULTS: At Week 24, the mean dose of glargine was 37.6 IU/day. There were significant decreases (p<0.0001) compared with baseline, for mean HbA(1c) (1.5%), mean FPG (88.3mg/dL) (p<0.0001), mean PPG (112.0mg/dL), and mean fasting C-peptide (1.14 ng/mL). Peptide index (peak basal/basal) in carbohydrate challenge test increased by 2.24 units. No severe adverse events, including severe hypoglycemia were reported. CONCLUSIONS: Our study suggests that combined therapy of insulin glargine and glimepiride should be considered for T2DM patients who have unsatisfactory response to previous OAD treatment. PMID- 17919764 TI - Left ventricular diastolic dysfunction in normotensive diabetic patients in various age strata. AB - Although patients with type 2 diabetes demonstrate cardiac diastolic dysfunction, it is well known that cardiac diastolic dysfunction is produced by hypertension and aging. The purpose of the present study was to elucidate the cardiac structure and function in normotensive patients with type 2 diabetes in various age strata in order to assess the effect of diabetes mellitus itself on cardiac function. Echocardiographic examination was performed in 77 normotensive patients with type 2 diabetes (age: 63+/-11 years) and 76 healthy control subjects (age: 60+/-11 years) who were in their forties, fifties, sixties, and seventies. The left ventricular (LV) wall thickness and dimension were measured by M-mode echocardiography. The relative wall thickness, LV mass index, and LV ejection fraction (LVEF) were calculated. LV diastolic function was assessed by the peak velocity of early rapid filling (E velocity) and the peak velocity of atrial filling (A velocity), and the ratio of E to A (E/A) using the transmitral flow velocity, which were obtained by Doppler echocardiography. No difference was observed in the relative wall thickness, LV mass index, or LVEF between the diabetic patients and control subjects in any of the age strata. The E/A ratio in the patients with type 2 diabetes was not different from that in the control subjects in their 40s (1.17+/-0.35 versus 1.20+/-0.36). However, E/A was significantly lower in the diabetic patients than in the control subjects in their 50s (0.87+/-0.28 versus 1.14+/-0.24), 60s (0.78+/-0.22 versus 0.97+/-0.27), and 70s (0.66+/-0.19 versus 0.84+/-0.21) (p<0.05). The duration of type 2 diabetes was significantly longer in patients in their 50s (7.0+/-2.5 years), 60s (8.0+/-3.2 years), and 70s (10.4+/-3.2 years) than in patients in their 40s (3.3+/-1.9 years) (p<0.001). The value of E/A in the diabetic patients correlated with the duration of type 2 diabetes (r=-0.62, p<0.001). These results indicate that cardiac diastolic dysfunction without LV systolic dysfunction in patients with well-controlled type 2 diabetes is related neither to hypertension nor LV hypertrophy, but rather to aging and the duration of type 2 diabetes. PMID- 17919765 TI - Hyperplastic islets observed in "reversed" NOD mice treated without hematopoietic cells. AB - At the onset of type 1 diabetes, most of the insulin-producing pancreatic beta cells are destroyed by effector cells, and therefore, the following two factors, at a minimum, are necessary for "reversing" hyperglycemia in autoimmune diabetes; depletion of effector cells and enhancement of beta cell regeneration. In this study, we tried a novel approach for "reversing" autoimmune diabetes in a murine model. Here we show that remission could be achieved with a combination therapy of a single injection of complete Freund's adjuvant (CFA) and a single intraperitoneal injection of a pancreatic beta cell line, MIN6N-9a, in recent onset diabetic NOD (non-obese diabetic) mice. Five out of seven mice (71%) receiving MIN6N-9a and CFA became normoglycemic within 120 days after treatment, whereas only two of nine (22%) receiving vehicle instead of MIN6N-9a achieved remission. Histological examination of pancreatic specimens from "reversed" mice showed decreased islet number, but each islet was markedly hyperplastic; being about six times larger than those from controls. Although it has been reported that hematopoietic cells such as splenocytes differentiate into insulin-producing cells and play a key role, our data indicate that they are not an absolute requirement for the "reversal" of autoimmune diabetes. PMID- 17919766 TI - An improved algorithm for vessel centerline tracking in coronary angiograms. AB - For automated visualization and quantification of artery diseases, the accurate determination of the arterial centerline is a prerequisite. Existing tracking based approaches usually suffer from the inaccuracy, inflexion and discontinuity in the extracted centerlines, and they may even fail in complicated situations. In this paper, an improved algorithm for coronary arterial centerline extraction is proposed, which incorporates a new tracking direction updating scheme, a self adaptive magnitude of linear extrapolation and a dynamic-size search window for matched filtering. A simulation study is conducted for the determination of the optimal weighting factor which is used to combine the geometrical topology information and intensity distribution information to obtain the proposed tracking direction. Synthetic and clinical examples, representing some difficult situations that may occur in coronary angiograms, are presented. Results show that the proposed algorithm outperforms the conventional methods. By adopting the proposed algorithm, centerlines are successfully extracted under these complicated situations, and with satisfactory accuracy. PMID- 17919768 TI - 3D lithographically fabricated nanoliter containers for drug delivery. AB - Lithographic patterning offers the possibility for precise structuring of drug delivery devices. The fabrication process can also facilitate the incorporation of advanced functionality for imaging, sensing, telemetry and actuation. However, a major limitation of present day lithographic fabrication is the inherent two dimensionality of the patterning process. We review a new approach to construct three dimensional (3D) patterned containers by lithographically patterning two dimensional (2D) templates with liquefiable hinges that spontaneously fold upon heating into hollow polyhedral containers. The containers have finite encapsulation volumes, can be made small enough to pass through a hypodermic needle, and the 3D profile of the containers facilitates enhanced diffusion with the surrounding medium as compared to reservoir systems fabricated in planar substrates. We compare the features of the containers to those of present day drug delivery systems. These features include ease of manufacture, versatility in size and shape, monodisperse porosity, ability for spatial manipulation and remote triggering to release drugs on-demand, the incorporation of electronic modules, cell encapsulation, biocompatibility and stability. We also review possible applications in drug delivery and cell encapsulation therapy (CET). The results summarized in this review suggest a new strategy to enable construction of "smart", three dimensional drug delivery systems using lithography. PMID- 17919769 TI - Retreatment with pemetrexed-based chemotherapy in malignant pleural mesothelioma (MPM): a second line treatment option. AB - The role of second line treatment in malignant pleural mesothelioma (MPM) is currently undefined. We present four cases of patients who had durable responses from pemetrexed-based chemotherapy who were retreated with a similar regimen upon progression of their mesothelioma. This case series explores the possible role of retreatment with pemetrexed-based chemotherapy as a second line therapeutic option in MPM. PMID- 17919770 TI - The status of the conditional evolutionarily stable strategy. AB - The conditional evolutionarily stable strategy (ESS) has proven to be a versatile tool for understanding the production of alternative phenotypes in response to environmental cues. Hence, we would expect the theoretical basis of the conditional strategy to be robust. However, Shuster and Wade have recently criticized the conditional ESS based on Gross's 1996 proposal that most alternative reproductive tactics are conditional and have evolved by 'status dependent selection.' We critically assess Gross's status-dependent selection model and Shuster and Wade's critique. We find shortcomings and misconceptions in both. We return to the findings of the strategic models behind the conditional ESS and demonstrate how environmental threshold models use a reaction norm approach and quantitative genetic theory to understand the evolution of conditional strategies. PMID- 17919771 TI - Mass extinction events and the plant fossil record. AB - Five mass extinction events have punctuated the geological record of marine invertebrate life. They are characterized by faunal extinction rates and magnitudes that far exceed those observed elsewhere in the geological record. Despite compelling evidence that these extinction events were probably driven by dramatic global environmental change, they were originally thought to have little macroecological or evolutionary consequence for terrestrial plants. New high resolution regional palaeoecological studies are beginning to challenge this orthodoxy, providing evidence for extensive ecological upheaval, high species level turnover and recovery intervals lasting millions of years. The challenge ahead is to establish the geographical extent of the ecological upheaval, because reconstructing the vegetation dynamics associated with these events will elucidate the role of floral change in faunal mass extinction and provide a better understanding of how plants have historically responded to global environmental change similar to that anticipated for our future. PMID- 17919772 TI - Allopurinol is the most common cause of Stevens-Johnson syndrome and toxic epidermal necrolysis in Europe and Israel. AB - BACKGROUND: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are rare severe cutaneous adverse reactions. OBJECTIVES: We sought to update knowledge on the causes of SJS or TEN with a focus on the rate of allopurinol associated cases and to identify risk factors for allopurinol-associated SJS or TEN. METHODS: We conducted a multinational case-control study. RESULTS: In all, 379 patients with severe cutaneous adverse reactions validated as SJS or TEN and 1505 matched hospitalized control subjects were enrolled. Allopurinol was the drug most frequently associated with SJS or TEN, with 66 exposed patients (17.4%) and 28 exposed control subjects (1.9%) (adjusted odds ratio = 18, 95% confidence interval: 11-32). Allopurinol use was greater than in a previous case-control European study. Daily doses equal to or greater than 200 mg were associated with a higher risk (adjusted odds ratio = 36, 95% confidence interval: 17-76) than lower doses (adjusted odds ratio = 3.0, 95% confidence interval: 1.1-8.4). The risk was restricted to short-term use (or=10 years of age) and used to estimate the base reproduction number (R(0)) and the herd immunity threshold (H). Most VZV infection occurred in childhood, but there was a wide variation in transmissibility, with R(0) ranging from 16.9 in the Netherlands to 3.3 in Italy. Herd immunity thresholds varied from 70% in Italy to 94% in the Netherlands. There are substantial differences in VZV sero-epidemiology within the European region, which will need to be taken into account in designing national policies regarding VZV vaccination. PMID- 17919789 TI - Hepatitis C virus envelope glycoprotein immunization of rodents elicits cross reactive neutralizing antibodies. AB - Neutralizing antibody responses elicited during infection generally confer protection from infection. Hepatitis C virus (HCV) encodes two glycoproteins E1 and E2 that are essential for virus entry and are the major target for neutralizing antibodies. To assess whether both glycoproteins are required for the generation of a neutralizing antibody response, rodents were immunized with a series of glycoproteins comprising full length and truncated versions. Guinea pigs immunized with HCV-1 genotype 1a E1E2p7, E1E2 or E2 generated high titer anti-glycoprotein antibody responses that neutralized the infectivity of HCVpp and HCVcc expressing gps of the same genotype as the immunizing antigen. Less potent neutralization of viruses bearing the genotype 2 strain J6 gps was observed. In contrast, immunized mice demonstrated reduced anti-gp antibody responses, consistent with their minimal neutralizing activity. Immunization with E2 alone was sufficient to induce a high titer response that neutralized HCV pseudoparticles (HCVpp) bearing diverse glycoproteins and cell culture grown HCV (HCVcc). The neutralization titer was reduced 3-fold by the presence of lipoproteins in human sera. Cross-competition of the guinea pig anti-E1E2 immune sera with a panel of epitope mapped anti-E2 monoclonal antibodies for binding E2 identified a series of epitopes within the N-terminal domain that may be immunogenic in the immunized rodents. These data demonstrate that recombinant E2 and E1E2 can induce polyclonal antibody responses with cross-reactive neutralizing activity, supporting the future development of prophylactic and therapeutic vaccines. PMID- 17919790 TI - Lead and polycyclic aromatic hydrocarbons (PAHs) in surface soil from day care centres in the city of Bergen, Norway. AB - Surface soil (0-2cm) quality in 87 day care centres in the city of Bergen, Norway has been studied. Approximately 45% of the day care centres contained Pb and PAH values above recommended action levels. There are clear variations between different areas of the city. The old central part of the city hosts most of the contaminated day care centres. In suburban areas most of the day care centres have Pb and PAH concentrations below action levels. City fires, gas work emission, lead-based paint, and traffic are probably important anthropogenic contamination sources, together with uncontrolled transportation of soil from contaminated to clean areas. Geological or other natural sources are probably not an important contributor to the high levels of lead and PAH. PMID- 17919791 TI - Database search for safety information on cosmetic ingredients. AB - Ethical considerations with respect to experimental animal use and regulatory testing are worldwide under heavy discussion and are, in certain cases, taken up in legislative measures. The most explicit example is the European cosmetic legislation, establishing a testing ban on finished cosmetic products since 11 September 2004 and enforcing that the safety of a cosmetic product is assessed by taking into consideration "the general toxicological profile of the ingredients, their chemical structure and their level of exposure" (OJ L151, 32-37, 23 June 1993; OJ L066, 26-35, 11 March 2003). Therefore the availability of referenced and reliable information on cosmetic ingredients becomes a dire necessity. Given the high-speed progress of the World Wide Web services and the concurrent drastic increase in free access to information, identification of relevant data sources and evaluation of the scientific value and quality of the retrieved data, are crucial. Based upon own practical experience, a survey is put together of freely and commercially available data sources with their individual description, field of application, benefits and drawbacks. It should be mentioned that the search strategies described are equally useful as a starting point for any quest for safety data on chemicals or chemical-related substances in general. PMID- 17919792 TI - EU legislations affecting safety data availability of cosmetic ingredients. AB - With the introduction of the 6th and 7th Amendments (OJ L151, 32-37, 23 June 1993; OJ L066, 26-35, 11 March 2003) to the Cosmetic Products Directive (OJ L262, 169-200, 27 September 1976), imposing a testing and marketing ban on cosmetic products tested on animals, the retrieval of toxicological data on individual ingredients became of greater need. Since the majority of cosmetic ingredients are used for many other purposes than their cosmetic function, they fall under the scope of more than one EU Directive. An overview is given of EU legislation that could potentially affect the availability and interpretation of cosmetic safety data. It will become clear that, although cosmetics are regulated by a specific so-called "vertical" legislation, "horizontal" influences from other products' legislations play a role since they determine the type and amount of data that theoretically could be found on the specific substances they regulate. This knowledge is necessary while performing extended searches in databases and becomes indispensable when initiating negotiations with manufacturers or suppliers for obtaining the safety data required. PMID- 17919793 TI - How to improve skin notation. Position paper from a workshop. AB - The ICOH Scientific Committee on Occupational and Environmental Dermatoses organized an International Workshop on "Dermal risk assessment at workplace" with the aim of focussing on the different ways of approaching the concept of skin notation (S) for chemicals. The Workshop participants presented their ideas on several aspects of S such as the problems related to the absorption through the compromised skin, the different approaches to S and models that can be used as alternatives to S. Participants agreed to produce a position paper with the goal of exploring the actions needed to improve the S system towards international harmonization. They consider that further discussions are needed to obtain an international consensus, but at the same time they believe that by improving and harmonizing systems for setting S we can make an important contribution to improving health of people with potential dermal exposure to chemicals at work. PMID- 17919794 TI - Paid to share: IVF patients, eggs and stem cell research. AB - Following a recent decision by the human fertilisation and embryology authority (HFEA), British women undergoing in vitro fertilisation (IVF) treatment can be 'paid to share' their eggs with stem cell researchers. The HFEA and the clinic proposing the scheme present this as a 'win-win' arrangement benefiting both infertile women and couples and British science. It is also represented as concurrently both 'business as usual' and an exceptional case. Constituting a significant departure from the previous policy and practice of altruistic donation, the scheme has raised significant concerns among clinicians and activists. Here, we ask what questions feminists can bring to these debates without resorting to a position of either refusal or affirmation. Drawing on diverse materials from public debates, as well as social scientific literature on gamete and embryo donation, we undertake a close analysis of the discursive framing and justification of the proposal. We argue that these discourses are characterised by three linked areas of elision and distinction: treatment and research; eggs and embryos; and donation and selling. Our analysis highlights the need for innovative social, ethical and political consideration of egg sharing for stem cell research. PMID- 17919795 TI - From health research to social research: privacy, methods, approaches. AB - Information-rich environments in Canada, Australia, and the United Kingdom have been built using record linkage techniques with population-based health insurance systems and longitudinal administrative data. This paper discusses the issues in extending population-based administrative data from health to additional topics more generally connected with well being. The scope of work associated with a multi-faceted American survey, the Panel Study in Income Dynamics (PSID), is compared with that of the administrative data in Manitoba, Canada. Both the PSID and the Manitoba database go back over 30 years, include families, and have good information on residential location. The PSID has emphasized research design to maximize the opportunities associated with expensive primary data collection. Information-rich environments such as that in Manitoba depend on registries and record linkage to increase the range of variables available for analysis. Using new databases on education and income assistance to provide information on the whole Manitoba population has involved linking files while preserving privacy, scaling educational achievement, assessing exposure to a given neighborhood, and measuring family circumstances. Questions being studied concern the role of the socioeconomic gradient and infant health in child development, the comparative influence of family and neighborhood in later well being, and the long-term effects of poverty reduction. Issues of organization of research, gaps in the data, and productivity are discussed. PMID- 17919796 TI - Repeat suicide attempts in Hong Kong community adolescents. AB - It has been well documented that a history of suicide attempts confers risk for subsequent attempts; however, efforts to explain how variables may change after a previous attempt and in turn relate to future suicide attempts are rare in the literature. This study presents longitudinal data on adolescent suicide attempts in Hong Kong, and examines whether the data support the "crescendo" model to explain repeat suicide attempts. One thousand and ninety-nine community adolescents aged 12-18 years were evaluated at two assessment points 12 months apart (T1 and T2). The study assessed (1) risk factors at T1 for a suicide attempt between T1 and T2, (2) whether a suicide attempt during the 12 months prior to T1 predicted an attempt between the two assessment points, and (3) whether the indicators of distress worsened from T1 to T2 if an attempt had taken place in the interim. The results indicated that: (1) depressive symptoms, substance use, and suicidal ideation measured at T1 were independent predictors of a suicide attempt between T1 and T2; (2) suicide attempt in the year prior to T1 predicted suicide attempt between T1 and T2 after controlling for other predictors; and (3) suicide attempt between T1 and T2 was a predictive factor for a negative change from T1 to T2 in substance use, suicidal ideation, family relationships, depression, anxiety, and life stress. These findings are consistent with the "crescendo" model proposing that the risk of repeat attempts is enhanced following a previous suicide attempt. PMID- 17919797 TI - Differences in cytokines between non-suicidal patients and suicidal patients in major depression. AB - Several studies have shown that there is an imbalance between pro-inflammatory and anti-inflammatory cytokines in major depressive disorder (MDD). However, little is known about the role of cytokines in suicide. In the present study, amounts of IL-6, IL-2, IFN-gamma, IL-4, and TGF-beta1 produced by mitogen stimulated whole blood were measured in 36 MDD patients who had recently attempted suicide, 33 non-suicidal MDD patients, and 40 normal controls. The severity of depression symptoms and suicidal behaviors was evaluated using Hamilton's depression rating scale (HDRS), the Lethality Suicide Attempt Rating Scale (LSARS), and the Risk-Rescue Rating (RRR). Non-suicidal MDD patients had significantly higher IL-6 production than suicidal MDD patients and normal controls (p<0.001). Suicidal MDD patients had significantly lower IL-2 compared with non-suicidal patients and normal controls (p<0.001). Both MDD groups, with or without attempted suicide, had significantly lower IFN-gamma and IL-4 and higher TGF-beta1 production. HDRS scores had significant positive correlations with IL-6, IFN-gamma, and the Th1/Th2 ratio and significant negative correlations with IL-4 in non-suicidal depression patients (p<0.005); however, these correlations did not hold true for suicidal patients. Suicidal MDD patients had no significant correlations between the LSARS or RRR scores and cytokine release. Our findings suggest that the immune response has distinct differences between non-suicidal patients and suicidal patients. Non-suicidal MDD may be associated with increased IL-6 production and a Th1/Th2 imbalance with a shift to Th1, while suicidal MDD may be associated with decreased IL-2. PMID- 17919798 TI - New-onset diabetic ketoacidosis in a schizophrenic patient with multiple autoimmune disease during treatment with risperidone. PMID- 17919799 TI - Comments on "Association study of the serotonin transporter promoter polymorphism and mirtazapine antidepressant response in major depressive disorder". PMID- 17919800 TI - A dietary supplement for female sexual dysfunction, Avlimil, stimulates the growth of estrogen-dependent breast tumors (MCF-7) implanted in ovariectomized athymic nude mice. AB - Avlimil, a dietary supplement advertised to ameliorate female sexual dysfunction, is a mixture of eleven herbal components, and some herbal constituents of Avlimil (including black cohosh, licorice, red raspberry, red clover and kudzu) contain phenolic compounds, which are suggested to have estrogenic, anti-estrogenic, or androgenic potential for relieving menopausal symptoms. We hypothesize that Avlimil could modulate the growth of estrogen receptor positive human breast cancer (MCF-7) cells in vitro and in vivo. A dimethylsulfoxide (DMSO) extract of Avlimil (0.001-100 microg Avlimil powder equivalents/mL media) was tested for its estrogenic and anti-estrogenic effects on the growth of MCF-7 cells in vitro. We observed that the DMSO extract of Avlimil at low concentrations (0.1-50 microg/mL media) dose-dependently increased MCF-7 cell proliferation in vitro, and Avlimil DMSO extract at 100 microg/mL inhibited the growth of MCF-7 cells in vitro. Avlimil and some constituents (black cohosh and licorice roots) of Avlimil were fractionated by using sequential solvent extraction (hexane, ethyl acetate, and methanol) and the activities of the fractions were monitored by effects on the growth of MCF-7 cells. Depending on dosage (0.1-100 microg/mL media) both stimulatory and inhibitory effects of the extracts on the growth of MCF-7 cells were observed. The effect of dietary Avlimil at dosages approximating human intake was evaluated using ovariectomized mice implanted with MCF-7 cells. Animals were fed diets containing 500 ppm or 1000 ppm Avlimil for 16 weeks. Dietary Avlimil at 500 ppm stimulated MCF-7 tumors, but Avlimil at 1000 ppm had no apparent effect on the growth of MCF-7 tumors. The observation of stimulated tumor growth in the absence of uterine wet weight gains suggest that estrogenic/anti-estrogenic effects of Avlimil we observed may be dosage- and target tissue-specific and that Avlimil may not be safe for women with estrogen dependent breast cancer. The different biological effects of fractionated Avlimil components and the different concentration dependencies warrant further compound identification and dose-response studies, especially at recommended intake levels that could have estrogenic effects in women. PMID- 17919801 TI - Safety evaluation of chicken breast extract containing carnosine and anserine. AB - Chicken breast extract (CBEX) is obtained via hot water extraction of chicken breast and contains among its primary constituents carnosine and anserine, which are histidine-containing dipeptides present in the muscle tissues of most vertebrate species. Dietary intake of CBEX has been previously shown to buffer hydrogen ions formed during high-intensity exercise in human skeletal muscle cells, thereby inhibiting a decrease in muscle cell pH and subsequent muscle fatigue. The objective of this paper is to report the results of safety studies completed on CBEX. CBEX was determined to have an oral LD(50) value of more than 6000 mg/kg body weight in rats. Gavage doses of 500 or 2000 mg CBEX/kg body weight/day administered to rats for 90 days produced no toxicologically significant, dose-related, differences between control and treated animals with respect to body weight gain, food consumption, behavioral effects, hematological and clinical chemistry parameters, absolute and relative organ weights, or gross and microscopic findings. In the presence or absence of metabolic activation, CBEX exerted no mutagenic activity in the Ames assay conducted in various strains of Salmonella typhimurium and Escherichia coli. The results of these studies support the safety of CBEX as a potential dietary source of carnosine and anserine. PMID- 17919802 TI - Tumor angiogenesis--a potential target in cancer chemoprevention. AB - Tumor angiogenesis is critically important for the growth of solid tumors as tumors remain in dormant phase for a long time in the absence of the initiation of blood vessel formation. Tumors can grow up to approximately 2mm size without requirement of blood supply as diffusion is sufficient at this level to support the removal of wastes from and supply of nutrients to tumor cells. Therefore, angiogenesis process could be an important target to suppress tumor growth and metastasis. Angiogenesis is required at almost every step of tumor progression and metastasis, and tumor vasculature has been identified as strong prognostic marker for tumor grading. Endothelial cells are the main players of angiogenesis process and could be peculiar target for antiangiogenic therapy because they are non-transformed and easily accessible to achievable concentrations of antiangiogenic agents, and also are unlikely to acquire drug resistance. Several antiangiogenic strategies have been developed to inhibit tumor growth by targeting different components of tumor angiogenesis. Chemopreventive agents have been shown to target and inhibit different aspects and components of angiogenesis process and can be used conveniently as they are mostly non-toxic natural compounds and could be part of our daily diet. However, a risk assessment for the use of antiangiogenic phytochemicals is needed as they can also disrupt normal physiologic angiogenesis such as wound healing and endometrium development processes. This review focuses on how different chemopreventive phytochemicals target various components of angiogenesis, including angiogenic signaling, which usually starts from tumor cells producing angiogenic factors and affecting endothelial cells growth, migration and capillary vessel organization for tumor angiogenesis. PMID- 17919803 TI - Protection against hydrogen peroxide induced oxidative damage in rat erythrocytes by Mangifera indica L. peel extract. AB - Phytochemicals such as polyphenols and carotenoids are gaining importance because of their contribution to human health and their multiple biological effects such as antioxidant, antimutagenic, anticarcinogenic and cytoprotective activities and other therapeutic properties. Mango peel is a major by-product in pulp industry and it contains various bioactive compounds like polyphenols, carotenoids and others. In the present study, the protective effect of peel extracts of unripe and ripe mango fruits of two varieties namely, Raspuri and Badami on hydrogen peroxide induced hemolysis, lipid peroxidation, degradation of membrane proteins and its morphological changes are reported. The oxidative hemolysis of rat erythrocytes by hydrogen peroxide was inhibited by mango peel extract in a dose dependent manner. The IC(50) value for lipid peroxidation inhibition on erythrocyte ghost membrane was found to be in the range of 4.5-19.3 microg gallic acid equivalents. The mango peel extract showed protection against membrane protein degradation caused by hydrogen peroxide. Morphological changes to erythrocyte membrane caused by hydrogen peroxide were protected by mango peel extract. The results demonstrated that mango peel extracts protected erythrocytes against oxidative stress and may impart health benefits and it could be used as a valuable food ingredient or a nutraceutical product. PMID- 17919805 TI - The perception of invariant speech features in children with autism. AB - We investigated whether the good pitch-discrimination abilities reported in individuals with autism have adverse effects on their speech perception by compromising their ability to extract invariant phonetic features from speech input. The MMN, a brain response reflecting sound-discrimination processes, was recorded from children with autism and their controls for phoneme-category and pitch changes in speech stimuli under two different conditions: (a) when all the other features of the standard and deviant stimuli were kept constant, and (b) when constant variation with respect to an irrelevant feature was introduced to the standard and deviant stimuli. Children with autism had enhanced MMNs for pitch changes in both conditions, as well as for phoneme-category changes in the constant-feature condition. However, when the phoneme-category changes occurred in phonemes having pitch variation, the MMN enhancement was abolished in autistic children. This suggests that children with autism lose their advantage in phoneme discrimination when the context of the stimuli is speech-like and requires abstracting invariant speech features from varying input. PMID- 17919804 TI - Fetal responses to induced maternal relaxation during pregnancy. AB - Fetal responses to induced maternal relaxation during the 32nd week of pregnancy were recorded in 100 maternal-fetal pairs using a digitized data collection system. The 18-min guided imagery relaxation manipulation generated significant changes in maternal heart rate, skin conductance, respiration period, and respiratory sinus arrhythmia. Significant alterations in fetal neurobehavior were observed, including decreased fetal heart rate (FHR), increased FHR variability, suppression of fetal motor activity (FM), and increased FM-FHR coupling. Attribution of the two fetal cardiac responses to the guided imagery procedure itself, as opposed to simple rest or recumbency, is tempered by the observed pattern of response. Evaluation of correspondence between changes within individual maternal-fetal pairs revealed significant associations between maternal autonomic measures and fetal cardiac patterns, lower umbilical and uterine artery resistance and increased FHR variability, and declining salivary cortisol and FM activity. Potential mechanisms that may mediate the observed results are discussed. PMID- 17919806 TI - Preferences for symmetry in faces change across the menstrual cycle. AB - Symmetry in human male faces may be a cue to heritable fitness benefits and is found attractive. Preferences for facial masculinity, another proposed marker of genetic quality, have been found to vary in ways that may maximise evolutionary relevant benefits and masculinity is found to be of increased attractiveness at peak fertility across the menstrual cycle. Here we show that women prefer more symmetric faces at peak fertility (Study 1) and that such shifting preferences may be potentially strategic preferences as we found them to occur only for judgements concerning short-term relations and when women already had a partner (Study 2). Such preferences potentially indicate a strategy that maximises the quality of extra-pair/short-term partners or a quality dependent response to hormones. Such strategic preferences for symmetry may support the role of symmetry in signalling potential good-gene benefits. PMID- 17919807 TI - Challenges of cancer biomarker profiling. AB - OBJECTIVES: New biomarkers are being developed to identify individuals at risk for cancer, detect disease earlier, determine prognosis, detect recurrence, predict response to particular agents, and monitor response to treatment. This article attempts to address some of the challenges facing the research and medical communities in the delivery of new biomarkers for individualized medicine. METHODS: A variety of issues and barriers can affect the transfer of clinical tests from research to clinical practice. Differences in sample collection, handling or storage, and profiling techniques may influence the protein profile obtained by any method. RESULTS: Standard procedures and quality check schemes are necessary because there is a lack of definition to guarantee reproducibility of new procedures. From technical and economic viewpoints, the assay has to be sufficiently robust to be completed in community-based hospitals. Although traditionally cancer patients were treated with drugs of low toxicity or of high tolerance regardless of their efficacy in a given patient if the benefits of that drug are proven in both experimental and clinical conditions, recent advances have provided opportunities to adapt "tailored" treatment modalities. The evolving trend is the usage of patterns of markers instead of a single marker. Further challenges in biomarker development are in finding the relevant markers that have the right degree of specificity and sensitivity and a reliable test to measure the outcome. CONCLUSIONS: Discovery, testing, and validation of clinically appropriate and commercially useful tumor markers should permit individualization of therapy. PMID- 17919809 TI - Multi-timescale event-scheduling in multi-agent immune simulation models. AB - Multi-agent (or MA) -based design approaches have received much research attention lately for modeling immunological systems due to their efficacy in representing non-heterogeneous behaviors in the population under dynamic environmental and topological conditions. The update scheme of a MA model refers to the frequency of agent state updates and how these are related in temporal order. In contrast to verifiable agent behavioral rules at the individual level, the update scheme is a design decision made by the model developer at the systems level that is subject to realism and computational efficiency issues that directly affect the credibility and the usefulness of the simulation results. Previous works have mainly focused on the issue of realism with respect to synchrony of updates but have often overlooked the necessary heterogeneity in update frequencies due to multi-timescales in immunological phenomena. To incorporate such multi-timescales for realism, the efficiency of the update scheme arises as a nontrivial issue. An event-scheduling based asynchronous update scheme has the advantage of allowing arbitrary smaller timescales for realism and avoiding unnecessary execution and delays to achieve efficiency. In this paper we present the application of the event-scheduling update scheme to realistically model the B cell life cycle, and empirically compare its simulation performance with the widely adopted uniform time-step update scheme. The simulation results show a significantly reduced execution time (40 times faster) and also reveal the conditions where the event-scheduling update scheme is superior. PMID- 17919808 TI - Using matrix of thresholding partial correlation coefficients to infer regulatory network. AB - DNA arrays measure the expression levels for thousands of genes simultaneously under different conditions. These measurements reflect many aspects of the underlying biological processes. A method based on the matrix of thresholding partial correlation coefficients (MTPCC) is proposed for network inference from expression profiles. It includes three main parts: (1) hierarchical cluster analysis, (2) cluster boundaries establishment, and (3) regulatory network inference. The method was applied to the expression data of 2467 genes in Saccharomyces cerevisiae measured under 79 different conditions [Eisen, M.B., Spellman, P.T., Brown, P.O., Botstein, D., 1998. Cluster analysis and display of genome-wide expression patterns. Proc. Natl. Acad. Sci. 95, 14863-14868]. Using hierarchical clustering and cluster boundaries establishment, the 2467 genes were grouped into 12 clusters. The expression profiles of each cluster were expressed as a set of expression levels average over the cluster that constituted genes of each condition. Then the expression data of these clusters were subjected to the analysis of partial correlation, and the significance of each element in the obtained partial correlation coefficient matrix (PCCM) was examined by a permutation test. The corresponding undirected dependency graph (UDG) was obtained as a model of the regulatory network of S. cerevisiae. The veracity of the network was evidenced by the consistency of our results with the collected results from experimental studies. PMID- 17919810 TI - Somatostatin signaling and the regulation of growth and metabolism in fish. AB - The study of the somatostatins (SS) signaling system in fish has provided important information about the structure, function, and evolution of SSs and their receptors. The SS signaling system elicits widespread biological actions via multiple hormone variants, numerous receptor subtypes, and a variety of signal transduction pathways. SSs alter growth via both direct and indirect actions, including inhibiting growth hormone release at the pituitary, decreasing hepatic GH sensitivity, and lowering plasma IGF-I levels. Metabolism also is significantly influenced by SSs. SSs stimulate the breakdown of energy stores and influences digestion, food intake, nutrient absorption, and food conversion both directly and through the modulation of other hormonal systems. The study of fish, which display a diversity of habitat types and life history forms, reveals that the SS signaling system helps regulate energy partitioning and integrate metabolism with growth and other biological processes. PMID- 17919811 TI - Quality of life after extratemporal epilepsy surgery: a prospective clinical study. AB - PURPOSE: The purpose of this prospective clinical study was to assess quality of life (QOL) and impact of seizure status on QOL in patients with extratemporal epilepsy after surgery. PATIENTS AND METHODS: Twenty-three consecutive patients who had been operated due to extratemporal epilepsy were included in this study. Quality of Life Inventory in Epilepsy-10 (QOLIE-10) questionnaire was completed by all patients before 6 months and 2 years after surgery. Results obtained from short- and long-term follow-up were compared to baseline. Furthermore, patients who were seizure-free since surgery and those who had seizure were also compared in terms of outcome in QOL after surgery. RESULTS: All patients showed significantly improved QOL in both short- and long-term follow-ups compared to preoperative status regardless of seizure status (p<0.001). Seizure-free patients showed better QOL than those of patients who continued to have seizure during postoperative period. Furthermore, improved QOL was correlated with seizure status and shorter duration of epilepsy (p=0.001). CONCLUSION: Our findings showed that improved QOL is related to postoperative seizure status. However, future clinical studies including larger population of patients with extratemporal epilepsy are required to elucidate the role of other factors. PMID- 17919813 TI - Coagulation of wood extractives in chemical pulp bleaching filtrate by cationic polyelectrolytes. AB - The purpose of this research was to investigate the effectiveness of short-chain cationic polyelectrolytes of different molecular weights and charge densities in reducing turbidity and selectively removing toxic wood extractives from chemical birch pulp filtrate. The effects of chemical type, dosage and temperature were of interest. An effective performance was achieved with a copolymer of acrylamide and methacrylate of medium molecular weight and medium charge density at 72 degrees C and pH 5-6. The dosage range optimum for reducing the turbidity was 102 142 mg/L. Up to 92% of the wood extractives was selectively removed. PMID- 17919814 TI - Dye adsorption behavior of Luffa cylindrica fibers. AB - Using natural Luffa cylindrica fibers as adsorbent removal of methylene blue dye from aqueous solutions at different temperatures and dye concentrations was investigated in this study. Thermodynamics and kinetics of adsorption were also investigated. The adsorption isotherms could be well defined with Langmuir model instead of Freundlich model. The thermodynamic parameters of methylene blue (MB) adsorption indicated that the adsorption is exothermic and spontaneous. The average MB adsorption capacity was found out as 49 mg/g and average BET surface area of fibers was calculated as 123 m(2)/g. PMID- 17919812 TI - Hypoxia enhances LPA-induced HIF-1alpha and VEGF expression: their inhibition by resveratrol. AB - Lysophosphatidic acid (LPA) is a bioactive phospholipid that is involved in various cellular events, including tumor invasion and metastasis. In the present study, we investigated the effects of LPA and hypoxia on HIF-1alpha and VEGF expression, as well as the effect of resveratrol on LPA and hypoxia-induced HIF 1alpha and VEGF expression and human ovarian cancer cell migration. Our results show that LPA treatment under hypoxia increases HIF-1alpha protein level, which leads to increased expression of VEGF protein and mRNA. These increases in HIF 1alpha and VEGF expression are dramatically attenuated by resveratrol. The underlying mechanism of inhibition of HIF-1alpha expression by resveratrol seems to be associated with both inactivation of p42/p44 MAPK and p70S6K, as well as enhanced degradation of HIF-1alpha protein, resulting in profound decrease in VEGF expression and cell migration. Collectively, these results show that LPA under hypoxic condition enhances cell migration through the sequential induction of HIF-1alpha and VEGF, and that this enhancement is efficiently blocked by resveratrol. PMID- 17919815 TI - Cyanobacteria and their toxins in Guanting Reservoir of Beijing, China. AB - The present study investigated the cyanobacteria and one family of their toxins microcystins (MCs) in Guanting Reservoir of Beijing, China. The dominant species in the cyanobacteria found in August and September of 2006 was Microcystis, which accounted 99% of total algal cells. The specific species of the Microcystis in the cyanobacteria included Microcystis ichthyobalbe, Microcystis novacekii, Microcystis botrys and Microcystis aeruginosa which had different ratios in different sites. The qualitative analysis by HPLC showed that two microcystins were contained in cyanobacteria and one microcystin was in water of the reservoir. The major microcystins were microcystin-RR (MC-RR) and microcystin-LR (MC-LR), but only MC-LR was detected in water. The quantitative analysis by HPLC indicated that the maximum concentrations of MC-RR and MC-LR contained in cyanobacteria were 0.74 and 0.41 mg/g dry weight, respectively. The maximum microcystin concentration in water was 1.15 microg/L and others were below 1 microg/L. PMID- 17919816 TI - Differential Bos taurus cattle response to Babesia bovis infection. AB - Bovine babesiosis is a tick-borne disease caused by Babesia spp. haemoprotozoans. The disease is of great importance at tick enzootic unstable areas and hampers cattle production in several developing countries. The available immunisation alternatives are pre-immunition and attenuated vaccines. Despite being efficient and protective, they are unsafe as they use cattle blood cells as inoculum and may potentially spread other diseases. Another alternative to help in babesiosis control would be the identification of genetically resistant cattle to Babesia bovis infection. The objective of this work was to phenotype cattle based on primary response against B. bovis infection. Two-hundred and forty half-sib Hereford and Aberdeen Angus heifers (120 animals from each breed), 12-18-month old naive cattle, originated from a tick-free area in Southern Brazil, were used in the experiment. Animals were monitored following an inoculation with 1x10(7)B. bovis parasitised erythrocytes. Results showed three different phenotypes: 1 'susceptible', animals with babesiosis clinical signs that received treatment to avoid death; 2-'intermediate', animals with clinical signs: parasitaemia, >or=21.5% reduction in packed cell volume (PCV) and increase in body temperature when compared to their pre-challenge physiological parameters, no specific treatment was needed as animals self recovered from the disease, and 3 'resistant', animals without clinical signs that showed B. bovis presence in blood smears, <21.5% PCV reductions, with little or no increase in body temperature and no need for babesiosis treatment. The frequencies of each phenotype were: 45.4, 26.7, and 27.9%, respectively, demonstrating the existence of phenotypic variation for B. bovis in Bos taurus cattle. PMID- 17919817 TI - Efficacy of a topically applied formulation of metaflumizone on cats against the adult cat flea, flea egg production and hatch, and adult flea emergence. AB - A spot-on metaflumizone formulation was evaluated to determine its adulticidal efficacy, effect upon egg production, and ovicidal activity when applied to flea infested cats. Eight male and eight female adult domestic shorthair cats were randomly assigned to either serve as non-treated controls or were treated topically with a minimum of 40mg/kg metaflumizone in single spot-on Day 0. On Days -2, 7, 14, 21, 28, 35, 42, 49, and 56, each cat was infested with approximately 100 unfed cat fleas, Ctenocephalides felis felis. On Days 1, 2, and 3, and at 48 and 72h after each post-treatment reinfestation, flea eggs were collected and counted. At approximately 72h after treatment or infestation, each cat was combed to remove and count live fleas. Egg viability was determined by examining hatched eggs after 5 days and adult emergence was determined 28 days after egg collection. Metaflumizone provided >/=99.6% efficacy against adult fleas from Days 3 to 45 following a single application. Following treatment, egg production fell by 51.6% within 24h and 99.2% within 48h. Following subsequent weekly infestations egg production from treated cats was negligible out to Day 38, with >/=99.5% reduction relative to non-treated cats. Where there were eggs to evaluate, metaflumizone treatment did not have any apparent effect on the hatching of eggs or on the development and emergence of adult fleas from the eggs produced by fleas from treated animals. PMID- 17919818 TI - Cooling extensive burns: sprayed coolants can improve initial cooling management a thermography-based study. AB - This pilot study was designed to verify whether the spraying of coolant improves initial cooling in extensive burns. The cooling effects of 1l of sprayed water and 5l of poured water (at 22 degrees C) were tested; 53 healthy participants were cooled for 15 min over 18% of their total body surface, twice. Thermographic imaging measured the loss of skin temperature and assessed the homogeneity of cooling. With sprayed coolant the mean decrease of skin temperature was significantly higher (p < 0.003) throughout the entire cooling period and more homogeneous for the first 9 min (p < 0.003), compared with poured coolant. Infrared tympanic thermometry estimated core body temperature; neither poured nor sprayed water caused hypothermia. Even with a fifth of the volume of poured water, sprayed water cooled more efficiently. Thus, we conclude that spraying of coolant improves initial management. PMID- 17919819 TI - Dental and skeletal changes during pressure garment use in facial burns: a systematic review. AB - OBJECTIVE: Concern exists regarding the combined effect of the burn itself in addition to the continuous use of the pressure garments on craniofacial structures. The purpose of this systematic review was to evaluate the available evidence regarding dental and skeletal changes produced during pressure garment use in facial burns. METHODS: Several electronic databases were screened for relevant articles and reference lists of pertinent articles were also hand searched. RESULTS: Only a two-part study had the minimal set of criteria sought. They found that children with partial face burns had only a slightly reduced inferior growth of the mandible and an increase protrusion of anterior teeth, which stay unchanged even after discontinuation of the pressure garment. The group of children with total face burns showed changes in the direction of the growth of the mandible, from a normal anterior inferior direction to a more inferior direction, and in the maxillary horizontal growth. A trend of returning toward normality was found thereafter. CONCLUSIONS: There is scarcity of studies reporting dentomaxillofacial effects after pressure garment use in facial burns. The level of evidence presented in these articles does not give support to any strong conclusion regarding the craniofacial effects of facial burn management. PMID- 17919820 TI - Toxic epidermal necrolysis: use of Biobrane or skin coverage reduces pain, improves mobilisation and decreases infection in elderly patients. AB - INTRODUCTION: Toxic epidermal necrolysis is a rare disease with high mortality due to generalised infection, sepsis or lung involvement, and requires discontinuation of all potentially triggering medications and intensive care in a specialised burn centre. Apart from wound care with antiseptics, wound coverage may be achieved with a skin substitute; treatments are compared with regard to infection, protein loss, re-epithelialisation and mortality. PATIENTS AND METHODS: Of 14 people with toxic epidermal necrolysis affecting >30% body surface area, eight received daily dressing changes using Lavasept nd six received wound coverage with Biobrane. Demographic data, SCORTEN score, mortality, visual-analog pain scale, mobilisation, time to re-epithelialisation, serum protein, albumin, C reactive protein and leukocytes, and body temperature were evaluated in all cases. RESULTS: Mean age of patients was 68.0+/-14.8 years, mean body surface area affected was 66.4%, median SCORTEN score was three and overall mortality was 36%. In the Biobrane ompared with the Lavasept control) group, mean pain was significantly reduced (2.9 versus 5.5 on the scale, p<0.05), mobilisation was significantly earlier (walking at 3 days versus 7 days, p=0.003), re epithelialisation was complete in 12.5 days versus 16 days, and at 9 days there was reduced decrease of serum proteins and significantly lower levels of C reactive protein and white cells (p<0.05). CONCLUSION: Early wound coverage with synthetic skin substitute such as Biobrane s beneficial compared with conservative antiseptic wound treatment, but mortality rate is not significantly different. PMID- 17919821 TI - Transient selective factor X deficiency in a severely burned child. PMID- 17919822 TI - Cost analysis of Jelonet versus Suprathel in the management of split-thickness skin graft donor sites. PMID- 17919823 TI - Treatments for metastatic melanoma: synthesis of evidence from randomized trials. AB - BACKGROUND: Advanced melanomas (non-resectable Stage-III/IV) are fatal, with few effective treatments. It remains unclear if other drugs offer improvements over the standard, dacarbazine. PURPOSE: We quantified objective response rates (Complete+Partial response) of dacarbazine versus comparators for advanced cutaneous melanoma. METHODS: We retrieved all head-to-head randomized controlled trials involving dacarbazine and other drugs/combinations. Two reviewers searched MEDLINE (1966-Jan 2006), EMBASE (1980-2006), CINAHL (1982-2006) and Cochrane library, then compared results. Differences were resolved through consensus. Rates were combined using random effects meta-analysis. chi2 tested heterogeneity; points from Jadad's method were assessed to examine study quality. RESULTS: We found 48 studies having 111 active treatment arms [24 with dacarbazine monotherapy (n=1390), 75 with dacarbazine combinations (n=4962), and 12 with non-dacarbazine treatments (n=783)] treating 7135 patients. Overall, study quality was poor. Response to dacarbazine monotherapy ranged between 5.3% and 28.0% (average 15.3%), OR=1.31, CI(95%): 1.06-1.61; N=3356. Partial responses comprised 73% of successes. Only adding interferons improved response rates (OR=1.69, CI(95%): 1.07-2.68, N=778) but survival duration was not significantly longer (P=0.32), and trials with larger sample sizes found lower success rates. All other treatments alone or in combination were ineffective P>0.05. CONCLUSIONS: Dacarbazine generally produces poor outcomes. Adding other therapies offers minimal clinical advantages (possibly with interferons). In general, study quality was poor and sample sizes were small. This meta-analysis highlights the unmet need for effective treatment options for advanced melanoma. PMID- 17919827 TI - Reversible reduction in dendritic spines in CA1 of rat and ground squirrel subjected to hypothermia-normothermia in vivo: A three-dimensional electron microscope study. AB - A study was made at electron microscope level of changes in the three-dimensional (3-D) morphology of dendritic spines and postsynaptic densities (PSDs) in CA1 of the hippocampus in ground squirrels, taken either at low temperature during hibernation (brain temperature 2-4 degrees C), or after warming and recovery to the normothermic state (34 degrees C). In addition, the morphology of PSDs and spines was measured in a non-hibernating mammal, rat, subjected to cooling at 2 degrees C at which time core rectal temperature was 15 degrees C, and then after warming to normothermic conditions. Significant differences were found in the proportion of thin and stubby spines, and shaft synapses in CA1 for rats and ground squirrels for normothermia compared with cooling or hibernation. Hypothermia induced a decrease in the proportion of thin spines, and an increase in stubby and shaft spines, but no change in the proportion of mushroom spines. The changes in redistribution of these three categories of spines in ground squirrel are more prominent than in rat. There were no significant differences in synapse density determined for ground squirrels or rats at normal compared with low temperature. Measurement of spine and PSD volume (for mushroom and thin spines) also showed no significant differences between the two functional states in either rats or ground squirrels, nor were there any differences in distances between neighboring synapses. Spinules on dendritic shafts were notable qualitatively during hibernation, but absent in normothermia. These data show that hypothermia results in morphological changes which are essentially similar in both a hibernating and a non-hibernating animal. PMID- 17919826 TI - Increased asynchronous release and aberrant calcium channel activation in amyloid precursor protein deficient neuromuscular synapses. AB - Despite the critical roles of the amyloid precursor protein (APP) in Alzheimer's disease pathogenesis, its physiological function remains poorly established. Our previous studies implicated a structural and functional activity of the APP family of proteins in the developing neuromuscular junction (NMJ). Here we performed comprehensive analyses of neurotransmission in mature neuromuscular synapse of APP deficient mice. We found that APP deletion led to reduced paired pulse facilitation and increased depression of synaptic transmission with repetitive stimulation. Readily releasable pool size and total releasable vesicles were not affected, but probability of release was significantly increased. Strikingly, the amount of asynchronous release, a measure sensitive to presynaptic calcium concentration, was dramatically increased, and pharmacological studies revealed that it was attributed to aberrant activation of N- and L-type Ca(2+) channels. We propose that APP modulates synaptic transmission at the NMJ by ensuring proper Ca(2+) channel function. PMID- 17919825 TI - Corticotropin releasing factor-1 receptor antagonist, CP-154,526, blocks the expression of ethanol-induced behavioral sensitization in DBA/2J mice. AB - RATIONALE: Manipulation of glucocorticoid receptor signaling has been shown to alter the acquisition and expression of ethanol-induced locomotor sensitization in mice. It is unknown if other components of the hypothalamic-pituitary-adrenal (HPA)-axis modulate locomotor sensitization resulting from repeated ethanol administration. In the present investigation, we determined if pretreatment with an i.p. injection of CP-154,526, a selective corticotropin releasing factor (CRF) type-1 receptor antagonist, would block the acquisition and/or expression of ethanol-induced locomotor sensitization in male DBA/2J mice. METHODS: To assess the role of the CRF1 receptor in the acquisition of behavioral sensitization, mice were pretreated with an i.p. injection of CP-154,526 30 min before each of 10 sensitizing i.p. injections of ethanol. To determine the role of the CRF1 receptor in modulating the expression of ethanol-induced sensitization, mice that had previously been sensitized to the locomotor stimulant effects of ethanol were pretreated with CP-154,526 30 min before an i.p. injection of ethanol on the test day. In a third study, ethanol-naive mice were pretreated with CP-154,526 30 min before an initial i.p. injection of ethanol to determine the combined effects of the CRF1 receptor antagonist and ethanol on locomotor activity. Blood ethanol concentrations were assessed at the termination of sensitization studies. RESULTS: Pretreatment with CP-154,526 blocked the expression of ethanol-induced locomotor sensitization in DBA/2J mice but did not prevent the acquisition of sensitization. The ability of CP-154,526 to block the expression of ethanol induced locomotor sensitization was not attributable to alterations in blood ethanol levels or possible sedative effects produced by the combined administration of CP-154,526 and ethanol. CONCLUSIONS: These data provide novel evidence that CRF1 receptor signaling modulates the expression of ethanol-induced locomotor sensitization, and add to a growing literature suggesting a role for neurochemicals and hormones associated with the HPA-axis in behavioral sensitization resulting from repeated exposure to drugs of abuse. PMID- 17919828 TI - Dehydroepiandrosterone in the nucleus accumbens is associated with early onset of depressive-behavior: a study in an animal model of childhood depression. AB - Although the monoamine theory of depression is well studied, regarding childhood depression it is poorly supported. Antidepressant treatments affecting the monoaminergic system fail to ameliorate childhood depression in the same manner that they affect adult depression. The present study used the Flinders sensitive line (FSL) rat, a well-investigated genetic animal model of depression and Sprague-Dawley (SD) rats as controls. We co-measured monoamines and dehydroepiandrosterone (DHEA) levels in the nucleus accumbens on postnatal day 1, in prepubertal rats (35 days), and adult rats (4 months) in order to examine developmental characteristics in the monoamine systems. The results suggest that there are different ontogenetic patterns of monoaminergic activity in FSL and SD rats. While monoamine levels were different only in adulthood, FSL rats exhibited lower DHEA levels already in prepubertal childhood. These differences may be relevant to the poor response to antidepressant drugs observed in depressed children and suggest DHEA as a new marker for childhood depression. PMID- 17919829 TI - Affective differences among daily tobacco users, occasional users, and non-users. AB - The current study (n=340 college students) examined individual differences in affective functioning among daily tobacco users, occasional users, and non-users. Tobacco use frequency in the past 6 months was positively associated with negative affect, affect lability, and rising reactivity while negatively associated with positive affect and falling reactivity. Multinomial logit analysis indicated that affect lability was positively associated with the likelihood of being either a non-user or a daily user relative to an occasional user. Positive affect and falling reactivity were positively associated and rising reactivity negatively associated with the likelihood of being a non-user relative to a daily user. The results indicate that daily tobacco use is associated with multiple indices of dysregulated affect. Daily tobacco use is associated with increased reactivity and decreased emotional stability coupled with decreased positive affect and a slowed rate of returning to baseline once aroused. Greater affect lability emerged as the primary affective difference between daily and occasional users. PMID- 17919830 TI - High prevalence rates of tobacco, alcohol and drug use in adolescents and young adults in France: results from the GAZEL Youth study. AB - BACKGROUND: Rates of substance use among adolescents have increased in the 1990s, however little is known about current patterns of substance use among youths entering adulthood. METHODS: We studied sex and age-specific rates of substance use (tobacco, alcohol, cannabis, other illicit drugs, inhalants and psychotropic medications) in a large sample of French youths aged 12-26 years (the GAZEL Youth study, n=1333). RESULTS: Prevalence rates of substance use were high and varied with age and sex. Tobacco, cannabis and polysubstance use were most frequent among 19-21 year-olds (regular tobacco use: 41.5% in males, 39.9% in females; regular cannabis use: respectively 23.9% and 10.9%; tobacco+alcohol+cannabis: respectively 9.9% and 4.6%). Regular alcohol use was most frequent among 22-26 year-olds (29.8% in males, 15.6% in females). Across successive birth cohorts, the age of initiation of tobacco and cannabis use decreased. Males were consistently more likely to use psychoactive substances than females (except for tobacco and psychotropic medications). CONCLUSIONS: Rates of substance abuse peak in late adolescence but remain high among a subgroup of young adults. Moreover, substance use initiation appears to be occurring at increasingly younger ages. PMID- 17919831 TI - A target role for mast cell in the prevention and therapy of hepatic fibrosis. AB - Hepatic fibrosis is a common pathological process of chronic hepatic disease. Despite the extensive studies, the pathophysiological mechanisms in hepatic fibrosis remain unclear. Mast cell has a variety of physiological and pathological functions through the production of heparin, histamine, neutrophil chemoattractants, immunoregulatory cytokines, and mast cell-specific serine proteases tryptase and chymase. The survival and proliferation of mast cell are dependent upon stem cell factor. More recently, the data have suggested that mast cell has been associated with hepatic fibrosis in many chronic liver diseases. However, to what extent the mast cell effects the hepatic fibrosis remains to be clarified. Several therapeutic approaches to inhibit mast cell activation have already demonstrated some clinical utility in tissue fibrosis or inflammatory diseases such as the use of mast cell stabilizers, inhibitors of tryptase or chymase, blockade of stem cell factor and anti-IgE therapy. The article introduces the hypothesis that mast cell has a central role when it is affected by its activation state in the progression of hepatic fibrosis, thus new therapeutic strategies for treatment of hepatic fibrosis are suggested by this hypothesis. Considering the important role of mast cell and the development of these tangible therapeutic approaches in hepatic fibrosis will enable us to target any types of chronic liver diseases, which appears to be a more reasonable or a promising strategy. PMID- 17919833 TI - Proposing a causal link between thyroid hormone resistance and primary autoimmune hypothyroidism. AB - Resistance to thyroid hormone (RTH) is a rare, inherited condition. It is characterised by raised circulating fT4 and TSH levels. The literature contains a number of descriptions of the finding of thyroid autoantibodies in patients with RTH. Until now, this has been attributed to the coincidental development of primary autoimmune thyroiditis as a second unrelated pathology. Our hypothesis is that the chronic TSH elevation in RTH stimulates lymphocytes to produce the pro inflammatory cytokine TNF-alpha. TNF-alpha, in turn mediates thyroid cell destruction by binding to its receptors on thyrocytes, or indirectly by potentiating antibody formation or cytotoxic T lymphocyte production. PMID- 17919832 TI - The venous hypothesis of hydrocephalus. AB - Pressure in the central nervous system (CNS) depends upon the volume of tissue that it contains. This includes blood, cerebrospinal fluid (CSF), nerves and any space occupying lesions. The dependency of pressure on volume arises because the CNS is confined by bone. Venous and CSF pressure is linked to overall pressure. Arterial pressure can increase in response to overall pressure to maintain arterial supply. Continuous arterial supply can be maintained because venous blood flows out of the CNS. Reduced volumes of arterial blood will enter the system if venous outflow is interrupted. Increase in CNS volume, as occurs with space occupying lesions, causes compression of veins. This may result in increased venous pressure and reduction in flow of blood out of the CNS. Cerebrospinal fluid (CSF) is extracellular fluid; its absorption back into the circulation is influenced by venous pressure. Any increased in CNS tissue volumes can therefore lead to CSF accumulation. This may then exacerbate the hydrocephalus by further increasing overall CNS volume. Free flow of CSF around the CNS facilitates venous drainage. Blockages to CSF flow can act like space occupying lesions. Chiari malformations, where the cerebellar tonsils obstruct the foramen magnum lead to reductions in CSF flow that can occur intermittently. This leads to impairment of venous drainage which may result in accumulation of CSF. The head or the spine can be affected together or separately. The manifestation of excess fluid accumulation is hydrocephalus and syringomyelia. The speed and origin of venous insufficiency influences the morphology of individual cases particularly with regard to lateral ventricle size. When pressure increases rapidly there may be little time for CSF accumulation. Oedema, compression of intracranial CSF spaces and cerebral ischaemia follows. When venous pressure is only slightly elevated CSF will accumulate and the manifestations of ischaemia may be less apparent, although ischaemia will be a feature of all instances of pathologically raised CNS pressure. PMID- 17919834 TI - Impact of image guidance on outcomes after external beam radiotherapy for localized prostate cancer. AB - PURPOSE: To verify whether rectal distention at the time of planning impacts outcomes in patients with localized prostate cancer treated with daily image guidance. METHODS AND MATERIALS: Between 1998 and 2002, a total of 488 prostate cancer patients were treated with intensity-modulated radiotherapy. The radiation dose was 70 Gy delivered at 2.5 Gy per fraction in all cases. All cases were treated with a 4-mm margin posteriorly. In all cases the total rectal volume documented on the CT scan was used for treatment planning. No special bowel preparation instructions were given, either for the simulation or the daily treatments. Before each daily treatment, alignment of the prostate was performed with the B-mode acquisition and targeting (BAT) transabdominal ultrasound system. The median follow-up for all 488 patients was 60 months (range, 24-96 months). RESULTS: For all patients the biochemical relapse-free survival (bRFS) rate at 5 years was 86%. The 5-year bRFS rate for the rectal distention<50 cm3, 50 to <100 cm3, and >or=100 cm3 groups was 90%, 83%, and 85%, respectively (p=0.18). To adjust for other potential variables affecting bRFS rates, a multivariate time-to failure analysis using the Cox proportional hazards model was performed. Rectal distention was not an independent predictor of biochemical failure on multivariate analysis (p=0.80). Rectal distention was not a predictor of rectal or urinary toxicity. CONCLUSION: The use of daily image guidance eliminates errors such as rectal distention at the initial planning stage that can affect outcomes after radiotherapy for localized prostate cancer. PMID- 17919836 TI - Pelvic insufficiency fracture after pelvic radiotherapy for cervical cancer: analysis of risk factors. AB - PURPOSE: To investigate the incidence, clinical characteristics, and risk factors of pelvic insufficiency fracture (PIF) after pelvic radiotherapy (RT) in cervical cancer. METHODS AND MATERIALS: Medical records and imaging studies, including bone scintigraphy, CT, and MRI of 557 patients with cervical cancer who received whole-pelvic RT between January 1998 and August 2005 were reviewed. RESULTS: Eighty-three patients were diagnosed as having PIF after pelvic RT. The 5-year cumulative incidence of PIF was 19.7%. The most commonly involved site was the sacroiliac joint. Pelvic pain developed in 48 patients (57.8%) at diagnosis. Eleven patients (13.3%) needed admission or narcotics because of severe pain, and others had good relief of symptoms with conservative management. In univariate analysis, age>or=55 years (p<0.001), anteroposterior/posteroanterior parallel opposing technique (p=0.001), curative treatment (p<0.001), and radiation dose>or=50.4 Gy (p=0.005) were the predisposing factors for development of PIF. Concurrent chemotherapy (p=0.78) was not significant. Multivariate analysis showed that age>or=55 years (p<0.001), body weight<55 kg (p=0.02), curative treatment (p=0.03), and radiation dose>or=50.4 Gy (p=0.04) were significant predisposing factors for development of PIF. CONCLUSION: The development of PIF is not rare after pelvic RT. The use of multibeam arrangements to reduce the volume and dose of irradiated pelvic bone can be helpful to minimize the risk of fracture, especially in elderly women with low body weight. PMID- 17919837 TI - Planning target margin calculations for prostate radiotherapy based on intrafraction and interfraction motion using four localization methods. AB - PURPOSE: To determine planning target volume (PTV) margins for prostate radiotherapy based on the internal margin (IM) (intrafractional motion) and the setup margin (SM) (interfractional motion) for four daily localization methods: skin marks (tattoo), pelvic bony anatomy (bone), intraprostatic gold seeds using a 5-mm action threshold, and using no threshold. METHODS AND MATERIALS: Forty prostate cancer patients were treated with external radiotherapy according to an online localization protocol using four intraprostatic gold seeds and electronic portal images (EPIs). Daily localization and treatment EPIs were obtained. These data allowed inter- and intrafractional analysis of prostate motion. The SM for the four daily localization methods and the IM were determined. RESULTS: A total of 1532 fractions were analyzed. Tattoo localization requires a SM of 6.8 mm left right (LR), 7.2 mm inferior-superior (IS), and 9.8 mm anterior-posterior (AP). Bone localization requires 3.1, 8.9, and 10.7 mm, respectively. The 5-mm threshold localization requires 4.0, 3.9, and 3.7 mm. No threshold localization requires 3.4, 3.2, and 3.2 mm. The intrafractional prostate motion requires an IM of 2.4 mm LR, 3.4 mm IS and AP. The PTV margin using the 5-mm threshold, including interobserver uncertainty, IM, and SM, is 4.8 mm LR, 5.4 mm IS, and 5.2 mm AP. CONCLUSIONS: Localization based on EPI with implanted gold seeds allows a large PTV margin reduction when compared with tattoo localization. Except for the LR direction, bony anatomy localization does not decrease the margins compared with tattoo localization. Intrafractional prostate motion is a limiting factor on margin reduction. PMID- 17919835 TI - Are increased tumor aneuploidy and heightened cell proliferation along with heterogeneity associated with patient outcome for carcinomas of the uterine cervix? A combined analysis of subjects treated in RTOG 9001 and a single institution trial. AB - PURPOSE: To look for possible associations between measurements of DNA index (DI), S-phase fraction (SPF), and tumor heterogeneity (TH) using flow cytometry and overall survival for patients with invasive cervical carcinoma treated with definitive irradiation. METHODS AND MATERIALS: A total of 57 patients with International Federation of Obstetrics and Gynecology Stages IB(2) through IVB cervical carcinomas treated with definitive radiotherapy with or without concurrent chemotherapy were enrolled into this registry study that involved flow cytometric analysis of fresh tissue from each cervical cancer obtained by pretreatment biopsy. These specimens were evaluated for DNA aneuploidy (DI 1.5), SPF (15%), and TH (uniploid vs. multiploid). RESULTS: In these analyses 27 of the patients were treated in Radiation Therapy Oncology Group protocol 9001, and an additional 30 were offered chemoradiation at a single institution. Forty-one patients had DI 1.5. Twenty nine patients had SPF 15%, and 2 had no determinable SPF. Forty three patients had uniploid and 14 multiploid tumors. The 4-year estimated overall survival rate for the entire study cohort was 62% (95% confidence interval 48%-74%). With a median follow-up of 3.7 years, there were no observable associations by univariate analysis for DI, SPF, or TH concerning patient survival. CONCLUSIONS: There were no statistically significant associations among DI, SPF, or TH and patient outcome. Additional studies are indicated to identify tumor biomarkers that could predict patients at risk for disseminated disease. PMID- 17919838 TI - Cyclooxygenase-2 expression as a predictor of para-aortic lymph node recurrence in uterine cervical cancer. AB - PURPOSE: The overexpression of cyclooxygenase-2 (COX-2) is associated with a worse prognosis and the development of distant metastases in cervical cancer. This matched-pair analysis examined whether COX-2 expression is associated with para-aortic lymph node (PALN) recurrence in uterine cervical cancer treated with radiotherapy (RT). METHODS AND MATERIALS: For this study, we matched 20 patients with PALN recurrence after definitive or postoperative RT by stage with 20 others who did not have PALN recurrence. Of the 20 patients with PALN recurrence, definitive or postoperative RT was performed in 11 and 9 patients, respectively. COX-2 expression was assessed immunohistochemically using a mouse monoclonal antibody on formalin-fixed paraffin-embedded tumor specimens taken before RT. A logistic regression model was used to predict for PALN recurrence. RESULTS: COX-2 was expressed in 28 (70%) of the 40 patients. The staining intensity was as follows: weak in 19 (47%), moderate in 6 (15%), and strong in 3 (8%) patients. The patients with PALN recurrence had much greater expression of COX-2 (17 patients, 85%) than did the control group (11 patients, 55%; p = 0.04). Strong staining intensity of COX-2 was seen only in the PALN recurrence group. The statistically significant factors associated with PALN recurrence were positive pelvic lymph nodes (odds ratio, 7.61; 95% confidence interval, 1.55-37.37; p = 0.01) and COX-2 expression (odds ratio, 1.47; 95% confidence interval, 1.04-2.09; p = 0.03). CONCLUSION: Our findings suggest that COX-2 overexpression in the initial tumor tissue might be associated with PALN recurrence after RT in cervical cancer patients. PMID- 17919839 TI - Accelerated hyperfractionated radiotherapy for cervical cancer: multi institutional prospective study of forum for nuclear cooperation in Asia among eight Asian countries. AB - PURPOSE: To evaluate the toxicity and efficacy of accelerated hyperfractionated radiotherapy (RT) for locally advanced cervical cancer. METHODS AND MATERIALS: A multi-institutional prospective single-arm study was conducted among eight Asian countries. Between 1999 and 2002, 120 patients (64 with Stage IIB and 56 with Stage IIIB) with squamous cell carcinoma of the cervix were treated with accelerated hyperfractionated RT. External beam RT consisted of 30 Gy to the whole pelvis, 1.5 Gy/fraction twice daily, followed by 20 Gy of pelvic RT with central shielding at a dose of 2-Gy fractions daily. A small bowel displacement device was used with the patient in the prone position. In addition to central shielding RT, intracavitary brachytherapy was started. Acute and late morbidities were graded according to the Radiation Therapy Oncology Group and Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer criteria. RESULTS: The median overall treatment time was 35 days. The median follow-up time for surviving patients was 4.7 years. The 5-year pelvic control and overall survival rate for all patients was 84% and 70%, respectively. The 5 year pelvic control and overall survival rate was 78% and 69% for tumors > or = 6 cm in diameter, respectively. No treatment-related death occurred. Grade 3-4 late toxicities of the small intestine, large intestine, and bladder were observed in 1, 1, and 2 patients, respectively. The 5-year actuarial rate of Grade 3-4 late toxicity at any site was 5%. CONCLUSION: The results of our study have shown that accelerated hyperfractionated RT achieved sufficient pelvic control and survival without increasing severe toxicity. This treatment could be feasible in those Asian countries where chemoradiotherapy is not available. PMID- 17919840 TI - Interval to biochemical failure highly prognostic for distant metastasis and prostate cancer-specific mortality after radiotherapy. AB - PURPOSE: Few biochemical parameters have been related to mortality. The present study examined the clinical utility of the interval to biochemical failure (IBF) as a prognostic factor for distant metastasis (DM) and prostate cancer-specific mortality (PCSM) after radiotherapy. METHODS AND MATERIALS: The study group consisted of 211 T1c-T3Nx-N0M0 patients who had experienced BF among 1,174 men treated with three-dimensional conformal radiotherapy alone. Biochemical failure was defined as a post-treatment prostate-specific antigen (PSA) level of at, or greater than, the PSA nadir plus 2 ng/mL. Cox proportional hazards modeling was used to identify independent predictors of DM and PCSM on multivariate analysis. RESULTS: An IBF of <18 months was independently predictive for DM (p = 0.008), as was a Gleason score of 7-10 (p = 0.0005), PSA nadir >or=2 ng/mL (p = 0.04), and decreasing radiation dose (p = 0.02) on multivariate analysis, including increasing pretreatment PSA level, PSA nadir >or=2.5 ng/mL, PSA doubling time of <3 months, and Stage T3 disease. An IBF of <18 months was the only predictor of PCSM (p = 0.0003) in the same model. The actuarial 5-year DM rate for an IBF of <18 vs. >or=18 months was 52% vs. 20% (p < 0.0001), and the actuarial PCSM rate was 36% vs. 6%, respectively (p = 0.0001). CONCLUSIONS: The IBF is an important descriptor of the PSA kinetics after radiotherapy to identify men at high risk of clinical failure and death. A IBF of <18 months could aid in selecting men for early, aggressive salvage therapy or participation in a clinical trial. PMID- 17919841 TI - Early or up-front radiotherapy improved survival of localized extranodal NK/T cell lymphoma, nasal-type in the upper aerodigestive tract. AB - PURPOSE: To investigate the role of early or up-front radiotherapy (RT), the optimal RT dose required to achieve appropriate treatment outcome and prognostic factors for patients with localized extranodal NK/T-cell lymphoma, nasal-type, in the upper aerodigestive tract. METHODS AND MATERIALS: Eighty-two patients were reviewed. Eight patients were treated with chemotherapy (CT) alone, 9 patients received RT alone, and 65 patients were given combined modality treatment of CT and RT (CMT). Of those 74 patients receiving RT, 31 patients were given up-front RT, whereas CT was the initial therapy for 43 patients and 41 of those 43 patients received early RT. RESULTS: Five-year overall survival (OS) and disease free survival (DFS) were 52.3% and 39.2%, respectively. RT was the only independent prognostic factor for both OS and DFS at both the univariate and multivariate level. The 5-year OS and DFS were better in patients receiving >or=54 Gy of RT as compared with that of <54 Gy (5-year OS 75.5% vs. 46.1%, p = 0.019; 5-year DFS 60.3% vs. 33.4%, p = 0.004). Up-front RT presented better survival in Stage I patients when compared with that of initial CT followed by early RT (5-year OS 90.0% vs. 48.9%, p = 0.012; 5-year DFS 78.7% vs. 39.9%, p = 0.021). CONCLUSION: Early or up-front RT had an essential role in improved OS and DFS in patients with localized extranodal NK/T-cell lymphoma, nasal-type, in the upper aerodigestive tract. The recommended tumor dose was at least 54 Gy. Up front RT may yield more benefits on survival in patients with Stage I disease. PMID- 17919843 TI - Clinical significance of margin status in postoperative radiotherapy for extremity and truncal soft-tissue sarcoma. AB - PURPOSE: To evaluate whether adjuvant radiotherapy (RT) in extremity and truncal soft-tissue sarcoma (STS) patients with microscopically positive or close margins after excision can achieve comparable local control to that of excision with negative margin plus RT. METHODS AND MATERIALS: A total of 150 patients (111 extremity and 39 trunk cases) treated with conserving surgery and adjuvant RT was analyzed. All surgical margins were classified as being a negative margin or a positive or close margin based on pathologic margin width. RT was delivered with a shrinking-field technique in 150 patients (median, 63 Gy). RESULTS: All patients were divided into two groups: (A) excision with negative margins plus RT (n = 56) and (B) excision with positive or close margins plus RT (n = 94). Overall, the 5-year local failure-free survival in all patients was 72.9%, and no significant differences were found between the two groups (Group A, 74.7%; Group B, 71.6%). High tumor grade was found to be a significant predictor of local failure. However, Group A was superior to Group B in distant metastasis-free survival (p = 0.02). No significant differences were shown in overall survival between the two groups. CONCLUSIONS: In our series, margin status did not predict for LF when adjuvant RT was used. We believe that when adjuvant RT is used, re resection may not be necessary for selected patients with positive or close pathologic margins in the management of extremity and truncal STS patients. PMID- 17919845 TI - Strategy for online correction of rotational organ motion for intensity-modulated radiotherapy of prostate cancer. AB - PURPOSE: To develop and evaluate a correction strategy for prostate rotation using gantry and collimator angle adjustments. METHODS AND MATERIALS: Gantry and collimator angle adjustments were used to correct for prostate rotation without rotating the table. A formula to partially correct for left-right (LR) rotations was derived through geometric analysis of rotation-induced clinical target volume (CTV) beam's-eye-view shape changes. For 10 prostate patients, intensity modulated radiotherapy (IMRT) plans with different margins were created. Simulating CTV LR rotation and correcting each beam by a collimator rotation, the corrected CTV dose was compared with the original and uncorrected dose. Effects of residual geometric uncertainties were assessed using a Monte Carlo technique. A large number of treatments representative for prostate patients were simulated. Dose probability histograms of the minimum CTV dose (D min) were derived, with and without online correction, resulting in a more realistic margin estimate. RESULTS: Dosimetric analysis of all IMRT plans showed that, with rotational correction and a 2-mm margin, D min was constant to within 3% for LR rotations up to +/-15 degrees . The Monte Carlo dose probability histograms showed that, with correction, a margin of 4 mm ensured that 90% of patients received a D min >or=95% of the prescribed dose. Without correction a margin of 6 mm was required. CONCLUSIONS: We developed and tested a practical method for (online) correction of prostate rotation, allowing safe and straightforward implementation of margin reduction and dose escalation. PMID- 17919842 TI - Involved-field, low-dose chemoradiotherapy for early-stage anal carcinoma. AB - PURPOSE: To report the results of patients with early-stage anal cancer treated using a low-dose, reduced-volume, involved-field chemoradiotherapy protocol. METHODS AND MATERIALS: Between June 2000 and June 2006, 21 patients were treated with external beam radiotherapy (30 Gy in 15 fractions within 3 weeks) and concurrent chemotherapy (bolus mitomycin-C 12 mg/m(2) on Day 1 to a maximum of 20 mg followed by infusion 5-fluorouracil 1,000 mg/m(2)/24 h on Days 1-4). Of the 21 patients, 18 underwent small-volume, involved-field radiotherapy and 3 were treated with anteroposterior-posteroanterior parallel-opposed pelvic fields. Of the 21 patients, 17 had had lesions that were excised with close (<1 mm) or involved margins, 1 had had microinvasive disease on biopsy, and 3 had had macroscopic tumor <2 cm in diameter (T1). All were considered to have Stage N0 disease radiologically. RESULTS: After a median follow-up of 42 months, only 1 patient (4.7%) had experienced local recurrence and has remained disease free after local excision. No distant recurrences or deaths occurred. Only 1 patient could not complete treatment (because of Grade 3 gastrointestinal toxicity). Grade 3-4 hematologic toxicity occurred in only 2 patients (9.5%). No significant late toxicity was identified. CONCLUSION: The results of our study have shown that for patients with anal carcinoma who have residual microscopic or very-small volume disease, a policy of low-dose, reduced-volume, involved-field chemoradiotherapy produces excellent local control and disease-free survival, with low rates of acute and late toxicity. PMID- 17919846 TI - [Is there a change in ecology of Candida species incriminated in fungemia? A 7 years study at Foch Hospital]. AB - Some Candidemia studies have documented changes in epidemiology of Candida species and some species were reported as emerging species. We conducted a study over a 6 years period and until 2005 we do not noticed any change in epidemiology of Candida even if Candida albicans still the most common species followed by Candida glabrata. No increase of candidemia was observed from 2000 to 2005 and we observed a decrease during the year 2006, this fact have to be confirm and may be related to other data: reinforcement of hygienic measures in our hospital, changes in treatment or preemptive treatment of yeasts and fungi with new azoles or candines molecules. On another side, patients from intensive care units and patients suffering of cancer were, as expected, the most represented population in our study. PMID- 17919844 TI - Randomized, multicenter, phase IIb study of preoperative chemoradiotherapy in T3 mid-distal rectal cancer: raltitrexed + oxaliplatin + radiotherapy versus cisplatin + 5-fluorouracil + radiotherapy. AB - PURPOSE: To prospectively compare the rates of pathologic response, acute toxicity, and sphincter preservation with two different schedules of preoperative chemoradiotherapy in patients with cT3 mid-distal rectal cancer. METHODS AND MATERIALS: Patients with cT3 and/or N+ resectable rectal carcinoma were randomized to receive one of the two following chemoradiotherapy regimens: cisplatin, 5-fluorouracil, and radiotherapy (PLAFUR) or raltitrexed, oxaliplatin, and radiotherapy (TOMOX-RT). For PLAFUR, cisplatin (60 mg/m(2)) was given on Days 1 and 29, with a prolonged infusion of 5-fluorouracil (1,000 mg/m(2)) on Days 1-4 and 29-32, plus concurrent radiotherapy (50.4 Gy in 1.8-Gy fractions daily). For TOMOX-RT, raltitrexed (3 mg/m(2)) and oxaliplatin (130 mg/m(2)) was given on Days 1, 19, and 38 with the same radiotherapy regimen as used for PLAFUR. Surgery was performed 6-8 weeks after completion of chemoradiotherapy. All pathologic specimens were reviewed by a designated expert pathologist. The primary endpoint of this study was pathologic tumor downstaging (defined as tumor regression grade 1-2). Secondary endpoints included the incidence of ypT0, clinical tumor downstaging, sphincter-saving surgery, and acute treatment-related toxicity. RESULTS: Between 2002 and 2005, 164 patients were accrued in 10 Italian centers, 83 patients in the PLAFUR arm and 81 in the TOMOX-RT arm. Overall, tumor regression grade 1-2 was observed in 76 patients (46.4%) and ypT0 in 49 (29.9%). The tumor regression grade 1-2 rate was 41.0% vs. 51.9% (p = 0.162) and the ypT0 rate was 24.1% vs. 35.8% (p = 0.102) for the PLAFUR vs. TOMOX-RT arm, respectively. The overall rate of tumor regression grade 1 and ypN+ was 4.6%. The occurrence of ypT downstaging was significantly greater in the TOMOX-RT arm (p = 0.035). Grade 3-4 acute toxicity occurred in 19 patients (11.6%): 7.1% in the PLAFUR arm vs. 16.4% in the TOMOX-RT arm. Sphincter-saving surgery was performed in 143 patients (87.2%) overall: 87.9% in the PLAFUR arm and 86.4% in the TOMOX RT arm. CONCLUSIONS: Compared with the PLAFUR regimen, TOMOX-RT achieved a greater incidence of downstaging but was associated with a correspondingly greater rate of acute Grade 3+ toxicity. With longer follow-up, the local control and survival rates might offer additional guidance as to the choice of regimen. PMID- 17919847 TI - Molecular diversity of the aminoterminal region of the G protein gene of human respiratory syncytial virus subgroup B. AB - Two major antigenic subgroups (designated A and B) have been described for human respiratory syncytial virus (HRSV). Between and within the two main subgroups, there is antigenic variation in the attachment protein G. The variability of the G protein is known to be located in two hypervariable regions of the ectodomain. Most investigators have studied the gene segment coding the C-terminal end of the protein, and little is known about the N-terminal variable region. In the present study, the genetic variability of HRSV subgroup B was evaluated by nucleotide sequencing of the N-terminal region of the G gene of 52 Tunisian isolates. Tunisian subgroup B isolates clustered into two main lineages designated arbitrarily as Tu-GB1 and Tu-GB2. Three distinct subtypes were identified within genotype Tu-GB2. The inter- and intragenotype nucleotide variability ranged from 4 to 8% and from 0 to 4%, respectively. Overall divergence values of the G sequences were inferior or equal to 15% at the aminoacid level. Comparison of sequences among Tunisian HRSV strains and viruses isolated in other geographical areas during different epidemics demonstrated close similarity to strains from Kenya, Belgium, the UK, Qatar, Canada and South Korea. PMID- 17919848 TI - Commodity specific rates of temporal discounting: does metabolic function underlie differences in rates of discounting? AB - Discounting rates vary as a function of commodity type. Previous studies suggest five potential characteristics of the commodity that could explain these differences: type of reinforcer (primary or secondary), if the commodity is perishable, if the commodity is satiable, if the commodity can be directly consumed, and immediacy of consumption. This paper suggests that these characteristics may best be viewed as related to a more fundamental characteristic: metabolic processing. In order to explore the possibility that metabolic processing underlies changes in discount rates, the difference in discounting between food, money, music CDs, DVDs, and books are compared. Music CDs, DVDs, and books share many characteristics in common with food, including gaining value through a physiological process, but are not directly metabolized. Results are consistent with previous findings of commodity specific discount rates and show that metabolic function plays a role in determining discount rates with those commodities that are metabolized being discounted at a higher rate. These results are interpreted as evidence that the discount rate for different commodities lies along a continuum with those that serve an exchange function rather than a direct function (money) anchoring the low end and those that serve a direct metabolic function capping the high end (food, alcohol, drugs). PMID- 17919849 TI - If everything is everywhere, do they share a common gene pool? AB - Marine phytoplanktons are highly dispersed with large population sizes and are often considered to be homogenous over their entire range. Thus, using this definition, one would predict that everything is everywhere for these microbes. However, recent molecular analyses have shown both spatial and temporal compartmentalisation in phytoplankton communities, thus calling into question the idea that everything is everywhere, especially if they do not share a global gene pool. Examples are present to document the range of biogeography that has been reported in the phytoplankton and a hypothesis as to how this relates to species evolution on a geological time scale is provided. PMID- 17919850 TI - Pre-, peri- and postnatal complications in Prader-Willi syndrome in a UK sample. AB - BACKGROUND: Few studies describe in detail the pregnancy and early development of infants with Prader-Willi syndrome (PWS). However, variations at these early stages may partially account for differences in the later phenotype. A recent paper described an abnormally high number of problems in pregnancy and early infancy in a large sample of people with PWS but this sample was not homogeneous with respect to age and potentially liable to cohort effects. AIMS: To describe the early development of infants with PWS, younger and more homogeneous for age, and to investigate whether such high rates of perinatal problems are still present despite medical advances and whether there are differences according to the genetic subtypes. STUDY DESIGN: Using a structured interview, data were collected from mothers and from relevant GP and hospital records. SUBJECTS: Forty six babies with PWS, born in a six-year period 2000-2006, and their mothers. OUTCOME MEASURES: Problems arising during pregnancy, the birth process and the neonatal period and the birth characteristics of the babies. RESULTS: An abnormally high number of problems associated with the early developmental period similar to those previously reported were observed. The only significant difference between the genetic subtypes was that mother's age was positively correlated with birth weight for UPD (and negatively correlated for deletion subtypes). CONCLUSIONS: High rates of, and variability in, the nature and severity of problems arising during early development have been confirmed. To establish whether variability in the later phenotype is influenced by such differences requires a longitudinal study. PMID- 17919851 TI - Editorial - Nutrition of the preterm infant. Best Practice Guidelines on nutrition of preterm infants. PMID- 17919852 TI - Matrix metalloproteinases in infants with posthemorrhagic hydrocephalus. AB - The cerebrospinal fluid matrix metalloproteinase (MMP) activities were measured in infants with posthemorrhagic hydrocephalus to elucidate the intrinsic mechanism for the resolution of ventricular dilation. Increased MMP-9 activities were observed in the patients who escaped a shunt operation, suggesting its potential contribution to the resolution of ventricular dilation. PMID- 17919853 TI - Growth curves for preterm infants. AB - The commonly used growth curves for preterm infants are four decades old and may not be suitable for the current population. Uncertainty exists regarding the most suitable curves for monitoring the growth of preterm infants. While intrauterine growth rate appears to be the ideal growth that needs to be attained by the preterm infants, it may not be feasible given the limitations set by the morbidities of prematurity. Babson and Benda's chart has been updated using recent data from large samples of preterm infants making it useful for monitoring growth of infants in the preterm period. Once a corrected age of 40 weeks is reached, the recently released WHO growth curves can be used to monitor their ongoing growth. While aiming for achieving intrauterine growth velocities in postnatal life, one should not lose sight of the potential short term adverse effects of aggressive nutrition and long term adverse effects of excessive catch up growth. PMID- 17919854 TI - Paraquat potentiates glutamate toxicity in immature cultures of cerebellar granule neurons. AB - Toxic concentrations of paraquat (0.2mM, 24h) caused death of both mature and immature cerebellar granule neurons (CGNs), which could be prevented by blockers of ionotropic glutamate receptors, or by removal of glutamine from cultural medium. Glutamate (Glu, 0.05-1mM, 24h) was highly toxic for mature CGNs while young CGNs were insensitive to the toxic effect of Glu. Measurements of the relative intracellular calcium ion concentration showed that the Glu-induced [Ca(2+)](i) rise in mature neurons was two times higher than that in young neurons. Subtoxic concentrations of paraquat did not affect the Glu-induced [Ca(2+)](i) rise in neurons, but lowered the CGNs survival only in immature cultures. These data provide evidence that oxidative stress induced by paraquat is a strong factor modulating the glutamate-induced damage to immature CGNs. PMID- 17919855 TI - Effect of insulin administration on ovarian function and estrus induction in acyclic goats. AB - The present experiment was conducted to study the effect of insulin on follicular development, ovarian steroid profiles and estrus induction in 14 non-descript acyclic goats. Experimental animals were randomly divided into two groups, i.e. group A (control, n=7) and group B (insulin-treated, n=7). The animals of treatment group were injected with long acting bovine insulin subcutaneously at 0.2IU/kg body weight once daily for five consecutive days. However animals of control group were injected with normal saline only as a placebo. Animals were subjected to detection of estrus twice daily (morning and evening) using an apronized buck and visual observation of estrus signs. Simultaneously animals of both groups were subjected to B-mode transrectal ovarian ultrasonsography for follicular development and blood collection on days 0, 4, 8, 12, 16, 20, 24 and 28 for the estimation of serum estradiol-17beta, progesterone and nitric oxide (NO) profiles, respectively. The results revealed significantly greater total number of follicles in insulin-treated goats as compared to control. The diameter of corpus luteum (CL) in insulin-administered goat was ranging between 5.14 and 6.14mm. Significantly higher concentrations of estradiol-17beta on days 4 and 8 and progesterone on days 12, 16, 20 and 24 were observed in insulin-treated goats as compared to control. Significantly higher concentration of NO was observed in insulin-treated group as compared to control. In the treatment group estrus was recorded in 5/7 goats (71%), with estrus duration of 19+/-2.6h at a mean interval of 189+/-17.7h from the initiation of treatment. Ovulation occurred in 4/5 (80%) of responding animals, as confirmed by detecting CL ultrasonographically. However in control none of the animal displayed signs of estrus during the observation period. The results indicate favorable effect of insulin on ovarian function. PMID- 17919856 TI - Spermatozoal morphologies and fructose and citric acid concentrations in agouti (Dasyprocta leporina) semen. AB - This study was conducted to identify the levels of fructose and citric acid, and sperm morphologies in agouti (Dasyprocta leporina) semen. These parameters may be important in identifying highly fertile semen from the agouti. The objectives were: (1) to investigate spermatozoal abnormalities in agouti semen and (2) to determine the concentrations of seminal fructose and citric acid in agouti semen samples. Semen samples were collected from 16 anaesthetised male agouti by electro-ejaculation. Fructose and citric acid concentrations were 256.86+/-63.54 mg/dl and 1877+/-147 mg/dl, respectively, measured with ELISA kits. Sperm morphologies, examined using eosin-negrosin staining, showed 11 morphologies. The most abundant (68.5%) sperm morphology (M1) showed no known sperm defects. Means for head, mid piece, tail and total length of the agouti spermatozoa was 5.23+/ 0.04 microm, 5.18+/-0.08 microm, 37.52+/-0.24 microm and 47.96+/-0.25 microm, respectively for M1 sperm. The means of spermatozoa head and mid piece width and semen volume were 3.26+/-0.04 microm, 0.70+/-0.02 microm and 0.47+/-0.16 ml, respectively. It was concluded that as the fructose concentration in agouti ejaculate increased the percentage of spermatozoa with known spermatozoa defects increased (r=0.506; P<0.037; n=32). It is suggested that the M1 sperm could be the most competitive spermatozoa in agouti ejaculate. In conclusion standards for identifying fertile agouti semen were established. PMID- 17919857 TI - Association between premenopausal progestogen use and postmenopausal hormonal therapy in the GAZEL cohort. AB - OBJECTIVES: The aim of this work was to verify the existence of and characterise the relationship between premenopausal progestogen use, subsequent use of estrogen-progestogen therapy (EPT) and risk factors for breast cancer, and to describe and analyse premenopausal progestogen use among women participating in a longitudinal study. METHODS: Data came from self-administered questionnaires mailed to 2254 pre- or peri-menopausal women aged 45 years or older participating in the French GAZEL cohort and followed for 10 years. Bivariate and multivariate analyses first examined the association between progestogen use and women's and physicians' characteristics. A generalised linear model was then used to analyse the association between progestogen use and EPT use, while taking into account factors associated with progestogen use. RESULTS: Thirty-six percent of the women had used progestogens before menopause: 56% of them 19-norpregnane derivatives, and 13% 17alpha hydroxyprogesterone derivatives. They were more likely to report breast pain, a family history of breast cancer and mood changes, to be thin, to consult a gynaecologist rather than a general practitioner, to consult a female physician, and to consult often. Of the 1756 women who had reached menopause, EPT users were more likely to have used progestogens premenopausally (adjusted RR: 1.2; 95% CI: 1.1-1.3). CONCLUSIONS: Physician characteristics play an important role in the use of progestogens before menopause. Our results also suggest that women using progestogens may have a higher risk of breast cancer, that is, that progestogen use may be a confounding rather than causal factor. The analyses concerning the relationship between EPT use and breast cancer risk must take progestogen use before menopause into account, as they do other confounding factors, such as age at menarche, parity, and use of oral contraceptives. PMID- 17919858 TI - Application of instrumental evaluation of color for the pre-formulation and formulation of rabeprazole. AB - The aims of this study were to fast screen the compatibility of rabeprazole and excipients using a spectrocolorimeter and to examine the relationship between the color change value and drug contents/drug degradation products in solid dosage forms. The color change values of rabeprazole-excipient mixtures were measured using a spectrocolorimeter, with six tablet formulations compressed using a single-punch instrumental tablet press. The rabeprazole and degradation products contents in the tablets were analyzed using an HPLC method, with the color change values of the tablets measured using spectrocolorimetery for 4 weeks. These experiments indicated that the instrumental evaluation of color was a speedy, simple and useful tool in the determination of the interaction between the drug and excipients, as well as in the formulation of solid dosage forms. The relationships of the % reduced drug contents versus the color change value, and those of the % drug degradation products versus the color change value were exponentially increased in formulations containing zinc stearate. On stress testing, the color change value of rabeprazole was inconsistent with previous reports, as the degradation of rabeprazole can be greatly influenced by humidity as well as temperature. Consequently, these results highlight the potential of color formation in the application of pre-formulation and formulation of drugs. PMID- 17919859 TI - Preparation of rapamycin-loaded chitosan/PLA nanoparticles for immunosuppression in corneal transplantation. AB - Rapamycin-loaded chitosan/polylactic acid nanoparticles with size of about 300 nm in diameter were prepared through nanoprecipitation method using cholesterol modified chitosan as a stabilizer. The surface coating of chitosan, which was demonstrated by zeta potential measurement, endowed the nanoparticles good retention ability at the procorneal area, facilitating the sustained release of rapamycin on the corneal. The immunosuppression in corneal transplantation of the nanoparticles was investigated using rabbit as animal model, the median survival time of the corneal allografts treated with nanoparticles was 27.2+/-1.03 days and 50% grafts still remained surviving by the end of the observation, while the group treated with 0.5% rapamycin suspension was 23.7+/-3.20 days. The median survival time of drug-free nanoparticles group and untreated groups were 10.9+/ 1.45 and 10.6+/-1.26 days, respectively. The results demonstrated the excellent immunosuppression of rapamycin-loaded chitosan/polylactic acid nanoparticles in corneal transplantation. PMID- 17919860 TI - Solid form selection of zwitterionic 5-HT4 receptor agonist. AB - From discovery synthesis of a zwitterionic pharmaceutical compound, 4-{[4-({[(3 isopropyl-2-oxo-2,3-dihydro-1H-benzimidazol-1-yl)carbonyl]amino}methyl)piperidin 1-yl]methyl}tetrahydro-2H-pyran-4-carboxylic acid (compound A), two anhydrous ZW I and ZW-II and two hydrate forms ZW-III and ZW-IV were identified. Although stable form ZW-I was chemically stable at 70 degrees C/75% RH for 10 days, it was transformed to hydrate form ZW-IV under ambient conditions within a few days, taking up water from atmospheric moisture. In order to select a solid form for further investigation, solid-state characterization, salt screening on 96-well plate, stable polymorph and hydrate screening and physical stability were performed. Based on the results of the salt screening, besylate, camsylate, hemi edisylate, hemifumarate, monosuccinate salts of compound A were prepared, and their polymorphism and chemical and physical stability were evaluated. From the viewpoint of stability and manufacturability, a stable form of besylate salt (BSA I), which had two anhydrous forms BSA-I and BSA-II and hydrate form BSA-III, was selected as a solid form. BSA-I was quite stable at high relative humidity and provided significant improvement of physical stability compared with ZW-I. PMID- 17919861 TI - Attachment of the utricular and saccular maculae to the temporal bone. AB - The present investigation concerns the true morphology of the attachment of the two otolith receptor organs the utricular and the saccular maculae in two and three dimensions. By applying a new visualization method, which utilized the application of X-ray microtomography and a method of contrast enhancement based on en-bloc staining in osmium tetroxide, we were able to overcome problems of artefact production such as tissue distortion and loss of valuable information that was present in previous studies. A series of more than 1000 axial sections were obtained for each of the specimens, which subsequently formed the basis for detailed 2D and 3D visualizations. Our interpretations of these data reveal that the saccular maculae are closely attached to the curved bony surface of the temporal bone as traditionally believed, but the utricular macula is attached to the temporal bone only at the anterior region of the macula. PMID- 17919862 TI - Consequences of unilateral hearing loss: time dependent regulation of protein synthesis in auditory brainstem nuclei. AB - Conductive hearing impairment results in marked changes in neuronal activity in the central auditory system, particularly in young animals [Tucci, D.L., Cant, N.B., Durham, D., 1999. Conductive hearing loss results in a decrease in central auditory system activity in the young gerbil. Laryngoscope 109, 1359-1371]. To better understand the effects of conductive hearing loss (CHL) on cellular metabolism, incorporation of (3)H-leucine was used as a measure of protein synthesis in immature postnatal day 21 gerbils subjected to either unilateral CHL by malleus removal or profound sensorineural hearing loss by cochlear ablation. (3)H-leucine uptake was measured after survival times of 6 or 48h. Protein synthesis values were standardized to measurements from the abducens nucleus and compared with measurements from sham animals at similar age/survival times. Protein synthesis in the medial superior olive (MSO) was found to be significantly down-regulated (bilaterally) after CHL in animals surviving 48h. However, 6h after CHL manipulation, protein synthesis is up-regulated in MSO (bilaterally) and in the ipsilateral medial nucleus of the trapezoid body. PMID- 17919863 TI - Temporal integration in absolute identification of musical pitch. AB - The effect of stimulus duration on absolute identification of musical pitch was measured in a single-interval 12-alternative forced-choice task. Stimuli consisted of pure tones selected randomly on each trial from a set of 60 logarithmically spaced musical note frequencies from 65.4 to 1975.5Hz (C2-B6). Stimulus durations were 5, 10, 25, 50, 100, and 1000ms. Six absolute-pitch musicians identified the pitch of pure tones without feedback, reference sounds, or practice trials. Results showed that a 5ms stimulus is sufficient for producing statistically significant above chance performance. Performance monotonically increased up to the longest duration tested (1000ms). Higher octave stimuli produced better performance, though the rate of improvement declined with increasing octave number. Normalization by the number of waveform cycles showed that 4cycles are sufficient for absolute-pitch identification. Restricting stimuli to a fixed-cycle waveform instead of a fixed-duration still produced monotonic improvements in performance as a function of stimulus octave, demonstrating that better performance at higher frequencies does not exclusively result from a larger number of waveform cycles. Several trends in the data were well predicted by an autocorrelation model of pitch extraction, though the model outperformed observed performance at short durations suggesting an inability to make optimal use of available periodicity information in very brief tones. PMID- 17919864 TI - Phytochemical profiling and phase II enzyme-inducing properties of Thunbergia laurifolia Lindl. (RC) extracts. AB - Thunbergia laurifolia Lindl. (Acanthaceae) or Rang Chuet (RC) is described in traditional medicine for protection against dietary and environmental toxicants. This work, therefore, investigated RC's phytochemical profile, antimutagenic activity, and xenobiotic detoxification potential in its extracts. RC extracts were prepared by infusion with water, ethanol, acetone and subsequently assayed for major phytochemical constituents. Total phenolic content was 24.33, 5.65, and 1.42microg gallic acid equivalent (GAE) per mL for water, ethanol and acetone extract, respectively. HPLC analysis identified caffeic acid and apigenin as primary constituents of water extracts. Acetone and ethanol extracts contained primarily chlorophyll a and b, pheophorbide a, pheophytin a, and lutein. Treatment of Hepa 1C1C7 cells with standardized RC extracts resulted in a dose dependent increase in QR specific activity for all extracts. Acetone extract (92microg GAE/mL) increased QR activity 2.8-fold, while ethanol (120microg GAE/mL) and water (1000microg GAE/mL) extracts increased QR activity by 1.35- and 1.56-fold, respectively. The RC extracts were subsequently assayed for mutagen and antimutagenic activity by bacterial reverse mutagenesis assay. All three RC extracts exhibited strong dose-dependent antimutagenic activity inhibiting 2 aminoanthracene induced mutagenesis up to 87% in Salmonella typhimurium TA 98. These results support the traditional medicinal use of RC for detoxification and suggest the potential role of both phenolic acids and natural chlorophyll constituents in modulating these effects. PMID- 17919865 TI - Screening of antifungal activity of plants from the northeast of Mexico. AB - Screening of the antifungal activity of plants from the northeast of Mexico against some of the main etiological agents inducing pulmonary mycoses, Candida albicans, Aspergillus fumigatus, Histoplasma capsulatum, and Coccidioides immitis, was conducted in vitro. We used the M27-A2 and M38-A methods from the National Committee for Clinical Laboratory Standards to screen the antifungal activity. Ten hydroalcoholic extracts from the 15 plants evaluated showed antifungal activity against at least one of these fungi. Following this, a differential extraction was conducted with solvents of different polarities, and 16 extracts showed activity ranging from 16 to 125microg/mL against the different fungi. Toxicity of the extracts was evaluated by means of the test of lethality to Artemia salina, two of which were shown to have toxicity. PMID- 17919867 TI - Experimental evaluation of rigor mortis IX. The influence of the breaking (mechanical solution) on the development of rigor mortis. AB - Objective measurements were carried out to study the possible re-establishment of rigor mortis on rats after "breaking" (mechanical solution). Our experiments showed that: *Cadaveric rigidity can re-establish after breaking. *A significant rigidity can reappear if the breaking occurs before the process is complete. *Rigidity will be considerably weaker after the breaking. *The time course of the intensity does not change in comparison to the controls: --the re-establishment begins immediately after the breaking; --maximal values are reached at the same time as in the controls; --the course of the resolution is the same as in the controls. PMID- 17919866 TI - A novel PCR-DGGE-based method for identifying plankton 16S rDNA for the diagnosis of drowning. AB - The diagnosis of drowning is one of the most difficult issues in forensic practice. We have developed a sensitive and specific PCR and DGGE method for identifying the 16S rDNA of plankton, which exists ubiquitously in all types of water. In order to evaluate the usefulness of this method for diagnosis of drowning, we used this method for detection of plankton 16S rDNA in drowned rabbits and non-drowned rabbits submerged after death, as well as two human drowning cases. Plankton DNA was identified from lung, liver, kidney, blood and brain of the drowned victims, and the DGGE patterns were helpful in indicating the site of drowning. Plankton DNA was also identified from two lung samples obtained from non-drowned rabbits. The results show that the new PCR-DGGE-based method is a potentially useful tool for diagnosing drowning. PMID- 17919868 TI - Non tuberculous mycobacteria infection of the parotid region: two familiar cases. AB - OBJECTIVE: We review two cases of non tuberculous mycobacteria infections of the parotid region in members of the same family. The parotid region represents a peculiar location for the disease and it is exceptional to discover cases in members of the same family. METHODS: Two unusual case reports of non tuberculous mycobacteria infections in two members of the same family are presented. We discuss the diagnostic criteria and review pertinent recent literature. To our knowledge, these are the first English language reports of NTM infections in member of the same family. RESULTS: We performed surgical exeresis of the lesions in parotid region together with the skin affected by the fistula; regular check ups for 24 months after surgery. CONCLUSIONS: Surgical exeresis of regional structures is the treatment for non tuberculous mycobacteria infections non responsive to antibiotic therapy. PMID- 17919870 TI - WITHDRAWN: Selection of a surrogate beta-lactam testing agent for initial susceptibility testing of doripenem, a new carbapenem. AB - This article has been withdrawn consistent with Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The Publisher apologizes for any inconvenience this may cause. PMID- 17919869 TI - Why cold water delays the onset of vestibular vertigo--an functional MRI study. AB - The mechanism of vertigo is unclear. Generally, the peak time or the latency of blood oxygenation level dependent (BOLD) effect is about 6s. However, clinically, the latency of vertigo or nystagmus induced by caloric stimulations is much longer than 6s, commonly about 30s induced by water of 30 degrees C or 44 degrees C. We hypothesize that there is an inhibitive power or mechanism for the occurrence of vestibular vertigo, since it is an unpleasant feeling. The caloric test was performed in healthy volunteers during the BOLD fMRI scanning. The overlaid results of statistical parametric mapping (SPM) showed that three brain regions showed neural activation during vestibular dizziness while deactivation occurred in response to cold water simulation: (1) supplementary motor area (SMA); (2) middle temporal area/medial superior temporal area (MT/MST); (3) visual association area (BA19). The time course of the regions further demonstrated that the signal decreased during the cold-water stimulation and increased during the period of vertigo. We therefore further hypothesize that there may be two forces for the production of vertigo: inhibitory power (IP) and promotive power (PP). The delayed onset of vertigo was the result of the interaction between IP and PP. All of our findings, for the first time, suggested such an original mechanism of vertigo. PMID- 17919871 TI - Use of lemon juice to increase crack cocaine solubility for intravenous use. PMID- 17919872 TI - Acute lower extremity pain in an adult patient secondary to bilateral popliteal cysts. AB - Lower leg and calf pain is a common presenting complaint in the Emergency Department and may involve a variety of etiologies, including musculoskeletal and vascular pathology such as deep venous thrombosis. Baker's cysts are a periodic finding in patients suspected of having deep venous thrombosis. As more emergency physicians perform their own lower extremity ultrasound examinations, they encounter Baker's cysts as the cause of unilateral leg pain or swelling. Bilateral Baker's cysts are very rare and may present a diagnostic challenge in the Emergency Department if bedside ultrasound is not available. We present a case of bilateral Baker's cysts in a young dancer who presented complaining of bilateral lower extremity pain and upper calf swelling. The bilateral cysts were diagnosed on bedside ultrasound by the treating emergency physician and a disposition quickly made. PMID- 17919874 TI - Chest pain in an adolescent with Marfan syndrome. PMID- 17919873 TI - Unenhanced CT in the diagnosis of small bowel intussusception. PMID- 17919875 TI - Can I ask you a question? PMID- 17919877 TI - PTEN and VEGF: possible predictors for sentinel lymph node micro-metastasis in breast cancer. AB - INTRODUCTION: The loss of PTEN expression and VEGF overexpression has been found to be correlated with metastasis in breast cancer patients. Despite significant advances in micro-metastasis detection methods, little is known about the relationship between micro-metastasis and primary tumors. The purpose of this study was to assess the association of VEGF and PTEN expression with micro metastasis in breast cancer. MATERIALS AND METHODS: As destination sites for micro-metastasis, we examined peripheral blood (BD), bone marrow (BM) and sentinel lymph node (SLN) from 53 breast cancer patients. Protein and gene expressions of VEGF and PTEN at the primary site were determined by immunohistochemistry (IHC). BD and BM samples were processed using immunocytochemistry (ICC). SLNs were examined by hematoxylin and eosin (H&E) staining and IHC. RESULTS: Percentages of the patients with micro-metastasis were 24.5% for BD, 56.6% for BM, 26.4% in SLN by H&E and 41.5% in SLN by IHC. VEGF overexpression was strongly correlated to loss of PTEN expression (P=0.001, r= 0.446). VEGF overexpression and loss of PTEN expression were significantly associated with SLN micro-metastasis by either H&E or IHC (P<0.001). On the contrary, there is no significant correlation between their expression and micro metastasis in BD and BM. CONCLUSIONS: Our results indicate possible value of using these biological markers to predict the risk of micro-metastasis in breast cancer. PMID- 17919876 TI - Patterns of ambulatory activity in subjects with and without intermittent claudication. AB - PURPOSE: This study compared the patterns of ambulatory activity in subjects with and without intermittent claudication. METHODS: The study participants were 98 subjects limited by intermittent claudication and 129 controls who were matched for age, gender, and race. Subjects were assessed on their ambulatory activity patterns for 1 week with a small, lightweight step activity monitor attached to the ankle using elastic Velcro (Velcro Industries BV, Manchester, NH) straps above the lateral malleolus of the right leg. The step activity monitor recorded the number of strides taken on a minute-to-minute basis, the time spent ambulating, and the time and number of strides measured at low (<15 strides/min), medium (15 to 30 strides/min), and high (>30 strides/min) cadences. RESULTS: Subjects with intermittent claudication took fewer total strides each day than the controls (3149 +/- 1557 strides/d vs 4230 +/- 1708 strides/d; P < .001) and fewer strides at medium (1228 +/- 660 strides/day vs 1638 +/- 724 strides/day; P = .001) and high (766 +/- 753 strides/day vs 1285 +/- 1029 strides/day; P < .001) cadences. Subjects with intermittent claudication also had a lower daily average cadence than the controls (11.8 +/- 2.9 strides/min vs 13.5 +/- 3.1 strides/min; P < .001) and spent less total time ambulating each day (264 +/- 109 min/day vs 312 +/- 96 min/day; P = .034), primarily at medium (58 +/- 30 min/day vs 75 +/- 32 min/day; P < .001) and at high (19 +/- 17 min/day vs 30 +/- 22 min/day; P = .001) cadences. CONCLUSION: Intermittent claudication is associated with lower total daily ambulatory activity owing both to less time ambulating and to fewer strides taken while ambulating, particularly at moderate and high cadences. PMID- 17919878 TI - Vibrio cholerae cytolysin is essential for high enterotoxicity and apoptosis induction produced by a cholera toxin gene-negative V. cholerae non-O1, non-O139 strain. AB - Cholera toxin (CT) gene-negative Vibrio cholerae non-O1, non-O139 strains may cause severe diarrhea though their pathogenic mechanism remains unclear. V. cholerae cytolysin (VCC) is a pore-forming exotoxin encoded in the hlyA gene of V. cholerae whose contribution to the pathogenesis is not fully understood. In this work, the virulence properties of a CT gene-negative V. cholerae non-O1, non O139 strain causing a cholera-like syndrome were analyzed. Inoculation of rabbit ileal loops with the wild type strain induced extensive fluid accumulation, accompanied by severe histopathological damage characterized by villus shortening, lymphangiectasia and focal areas of necrosis. These pathogenic effects were abrogated by mutation of the hlyA gene thus pointing out the main role of VCC in the virulence of the strain. Interestingly, this toxin was capable of triggering apoptosis in human intestinal cell lines due to its anion channel activity. Moreover, the wild type strain also induced increased apoptosis of the intestinal epithelium cells which was not observed upon inoculation of the VCC null mutant strain, indicating that VCC may trigger apoptotic cell death during infection in vivo. Altogether, these results support a main role of VCC in the pathogenesis of the CT gene-negative V. cholerae non-O1, non-O139 strain and identify apoptosis as a previously unrecognized cell death pathway triggered by VCC. PMID- 17919880 TI - Not every hospital death can be anticipated. PMID- 17919879 TI - Hispanic language version of the family evaluation of hospice care. PMID- 17919881 TI - Phototoxicity assessment of drugs and cosmetic products using E. coli. AB - A gram negative bacteria Escherichia coli (Dh5alpha strain) was developed as an alternate test system of phototoxicity. Eight drugs (antibiotics) and cosmetic products (eight face creams) were examined for their phototoxicity using this test system. Five known phototoxic compounds were used to validate the test system. UVA-radiation induced phototoxicity of these compounds was tested by agar gel diffusion assay. Decrease in colony forming units (CFU) was taken as an end point of phototoxicity. The phototoxic compounds and antibiotics produced significant reduction in CFU (p<0.001) at 80 microg/ml concentrations under exposure to UVA-radiation (5.4-10.8 J/cm(2)). One face cream was found phototoxic and produced significant decrease in CFU of E. coli at 1.0mg/ml concentration under UVA exposure (10.8 J/cm(2)). The minimum effective concentration of tetracycline and dose of UVA-radiation were also determined by observing growth inhibition of E. coli through disc diffusion assay. The observations suggested that E. coli can be used as an alternative test system for phototoxicity evaluation of chemicals. A battery of test systems is required to conclude the toxic/phototoxic potential of a chemical agent. In view of the speed, easiness, sensitivity and low cost, E. coli is introduced as one of the alternate test system for phototoxicity studies in safety evaluation of various chemical ingredients or formulations used in cosmetics and drugs. PMID- 17919882 TI - Component structure of the Repeatable Battery for the Assessment of Neuropsychological Status in dementia. AB - The Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) has been studied relatively extensively in normal samples, and its theoretically derived index scores have been demonstrated to be useful in the assessment of a variety of clinical conditions. However, examinations of the empirical relationships between individual subtests are limited. The intent of the present study was to explore the component structure of the instrument in a sample of 351 individuals with a diagnosed memory disorder, to examine the impact of demographic factors on these empirically derived components, and to explore differences in performance between diagnostic groups. Findings suggested a three component solution (Memory, Visuomotor Processing, and Verbal Processing). Demographic variables had relatively small, but significant relationships with various component scores. Significant differences were observed between probable Alzheimer's disease and non-Alzheimer's type dementia groups on the memory component score, but not on other component scores or on RBANS index scores. PMID- 17919883 TI - Response to Bigler (2007): the sky is not falling. PMID- 17919884 TI - Prediction of genetic risk for dyslipidemia. AB - The purpose of the present study was to identify genetic variants that confer susceptibility to dyslipidemia. A total of 5213 individuals from two independent populations were examined: Subject panel A comprised 3794 individuals who visited participating hospitals; subject panel B comprised 1419 community-dwelling elderly individuals. The genotypes for 100 polymorphisms of 65 candidate genes were determined. The chi(2) test and multivariable logistic regression analysis revealed that seven polymorphisms of APOA5, APOC3, APOA1, ACAT2, and LPL were significantly associated with hypertriglyceridemia, six polymorphisms of APOA5, LIPC, and CYP3A4 with low HDL-cholesterol, and three polymorphisms of APOE and CCR2 with high LDL-cholesterol in subject panel A. For validation of these associations, the same polymorphisms were examined in subject panel B. Six polymorphisms of APOA5, APOC3, APOA1, and LPL were again significantly associated with hypertriglyceridemia, three polymorphisms of APOA5 with low HDL-cholesterol, and two polymorphisms of APOE with high LDL-cholesterol. Serum triglyceride, HDL cholesterol, and LDL-cholesterol concentrations differed significantly among genotypes of these corresponding polymorphisms in both subject panels. These results indicate that polymorphisms of APOA5, APOC3, APOA1, and LPL are determinants of hypertriglyceridemia and that those of APOA5 and APOE are determinants of low HDL-cholesterol and high LDL-cholesterol, respectively, in Japanese individuals. PMID- 17919885 TI - A novel role of minocycline: attenuating morphine antinociceptive tolerance by inhibition of p38 MAPK in the activated spinal microglia. AB - We have previously demonstrated that activation of p38 mitogen-activated protein kinase (p38 MAPK) in the spinal microglia mediates morphine antinociceptive tolerance. Minocycline, a selective inhibitor of microglia activation, has been reported to attenuate peripheral inflammation-induced hyperalgesia by depressing p38 MAPK in the spinal microglia. The aim of the present study is to explore the effect of intrathecal minocycline on the development of morphine antinociceptive tolerance and p38 activation in the spinal microglia induced by chronic morphine treatment. Minocycline (20, 50 and 100 microg) was given intrathecally 30 min before each morphine (15 microg) administration for consecutive 7 days. It was shown that minocycline attenuated tolerance to morphine analgesia in a dose dependent manner. Minocycline administration (50 microg) which was initiated on day 4 followed by another 4 days administration partially reversed the established morphine antinociceptive tolerance. However, minocycline treatment which was started on day 8 followed by its administration for 4 more days failed to reverse the established morphine tolerance. Immunohistochemical analysis showed that chronic intrathecal morphine-induced activation of p38 MAPK in the spinal microglia. Minocycline at a dose that was shown to antagonize tolerance to morphine analgesia significantly inhibited the increase in p38 MAPK activation in the spinal microglia. To our knowledge, this is the first study to demonstrate that minocycline antagonizes morphine antinociceptive tolerance, possibly due to the inhibition of p38 activation in the spinal microglia. PMID- 17919886 TI - Real-time 3-dimensional echocardiography for follow-up of Waterston shunt. AB - A 37-year-old man presented with Eisenmenger's reaction. He had progressive cyanosis and signs of right-sided heart failure. In early childhood he was palliated with a Waterston shunt for cyanotic congenital heart disease. Two dimensional echocardiography provided only a rudimentary view of his heart's anatomy and hemodynamics. Real-time three-dimensional echocardiography (RT3DE) was superior in visualizing both the size and the shape of the large shunt. RT3DE easily identifies and quantifies a key pathogenic factor for pulmonary hypertension with irreversible vascular damage in such patients. If applied early, RT3DE may prove to be a valuable follow-up tool. PMID- 17919887 TI - Mechanical dysfunction in extreme QT prolongation. AB - BACKGROUND: Although cardiac mechanical dysfunction has not been reported in congenital long QT syndrome (LQTS), theoretically abnormal cardiac repolarization may confer a substrate for mechanical dysfunction via calcium overload during the prolonged plateau period. We report the first clinical descriptions of a patient who supports this hypothesis. METHODS: Clinical, electrocardiographic, and echocardiographic evaluations were performed on a newborn presenting to Milwaukee Children's Hospital and subsequently referred to Mayo Clinic's LQTS Clinic. Mutational analysis of the 10 known LQTS-susceptibility genes was performed using denaturing high performance liquid chromatography and DNA sequencing. RESULTS: Presenting with fetal bradycardia from 2:1 atrioventricular block, a male Caucasian infant manifested extreme QT prolongation (QT interval corrected for heart rate: 800 milliseconds), 2:1 atrioventricular conduction, and macroscopic T wave alternans after birth. At 6 weeks of age, the infant demonstrated 20 seconds of torsades de pointes. The family history was unremarkable, screening electrocardiograms of first-degree relatives revealed normal findings, and mutational analysis of all known LQTS-susceptibility genes was negative. Echocardiographically, the infant demonstrated globally abnormal systolic and diastolic dysfunction before the onset of arrhythmias. CONCLUSIONS: This infantile presentation of QT extremis and cardiac dysfunction provides support for the hypothesis of mechanical perturbations stemming from abnormal repolarization. Future studies are needed to investigate whether or not the typical patient with congenital LQTS exhibits any evidence for mechanical dysfunction. PMID- 17919888 TI - Complications in alveolar distraction osteogenesis of the atrophic mandible. AB - To improve the starting point for placement of dental implants, 45 patients suffering from atrophied edentulous mandibles, with a vertical height varying between 7.3 and 15.8mm, were treated by alveolar vertical distraction osteogenesis (VDO). The mean follow-up period was 3 years, ranging from 1 to 7 years. Associated complications, as occurred during instalment of the distractor device, VDO period and consolidation phase, and also after dental implant placement, were evaluated. Observed complications were: early fractures (2%), late fractures (17%), bleeding or haematoma (4%), infections (6%), skin perforation (2%), mucosal dehiscence (8%), sensory disturbances (28%), sagging chin (13%) and failure of dental implants (13%). In 10 patients 2 complications and in 1 patient 3 complications were monitored. All complications occurred in the first year. It is concluded that VDO to restore vertical bone height in patients with mandibular atrophy is a surgically delicate technique with a high risk of various complications. The likelihood of the most striking complication, namely fracture occurrence, increases with decreasing residual bone height. PMID- 17919889 TI - Effect of orthognathic surgery for class III correction on quality of life as measured by SF-36. AB - The purpose of this work was to evaluate the quality of life of patients with Angle's class III malocclusion submitted to orthognathic surgery. Twenty-nine patients of both sexes, ranging in age from 17 to 46 years, with Angle's class III malocclusion and indication for surgical treatment, were evaluated about 30 days before surgery and 6 months postoperatively. Surgery consisted of maxillary advancement or mandibular retrusion, or both. The generic SF-36 questionnaire was used to evaluate the following eight domains: functional capacity, physical aspects, pain, general health status, mental health, emotional aspects, social aspects and vitality. Descriptive and inferential statistical analyses were used to determine possible interactions between timing of evaluation, gender and type of surgery. With respect to physical and social aspects, a significant difference in outcomes was observed, with mean scores being higher after surgery regardless of gender or type of surgery. Regarding emotional aspects, an interaction effect was observed for timing and gender, with higher mean scores only being obtained for females after surgery. Orthognathic surgery had a positive impact on the quality of life of both male and female patients, improving physical and social aspects, and on that of female patients, improving emotional aspects. PMID- 17919892 TI - To my oncologist - an open letter from a patient at the end of follow-up. PMID- 17919891 TI - Preparation and characterization of PLGA particles for subcutaneous controlled drug release by membrane emulsification. AB - Uniformly sized microparticles of poly(D,L-lactic-co-glycolic) (PLGA) acid, with controllable median diameters within the size range 40-140 microm, were successfully prepared by membrane emulsification of an oil phase injected into an aqueous phase, followed by solvent removal. Initially, simple particles were produced as an oil in water emulsion, where dichloromethane (DCM) and PLGA were the oil phase and water with stabiliser was the continuous phase. The oil was injected into the aqueous phase through an array type microporous membrane, which has very regular pores equally spaced apart, and two different pore sizes were used: 20 and 40 microm in diameter. Shear was provided at the membrane surface, causing the drops to detach, by a simple paddle stirrer rotating above the membrane. Further tests involved the production of a primary water in oil emulsion, using a mechanical homogeniser, which was then subsequently injected into a water phase through the microporous membrane to form a water in oil in water emulsion. These tests used a water-soluble model drug (blue dextran) and encapsulation efficiencies of up to 100% were obtained for concentrations of 15% PLGA dissolved in the DCM and injected through a 40 microm membrane. Solidification of the PLGA particles was followed by removal of the DCM through the surrounding aqueous continuous phase. Different PLGA concentrations, particle size and osmotic pressures were considered in order to find their effect on encapsulation efficiency. Osmotic pressure was varied by changing the salt concentration in the external aqueous phase whilst maintaining a constant internal aqueous phase salt concentration. Osmotic pressure was found to be a significant factor on the resulting particle structure, for the tests conducted at lower PLGA concentrations (10% and 5% PLGA). The PLGA concentration and particle size distribution influence the time to complete the solidification stage and a slow solidification, formed by stirring gently overnight, provided the most monosized particles and highest encapsulation efficiency. PMID- 17919890 TI - Subjective and objective quality of life in schizophrenia. AB - OBJECTIVE: Quality of life (QOL) is considered an important outcome in the treatment of schizophrenia, but the determinants of QOL are poorly understood in this population. Furthermore, previous studies have relied on combined measures of subjective QOL (usually defined as life satisfaction) and objective QOL (usually defined as participation in activities and relationships). We examined separately the clinical, functional, and cognitive predictors of subjective and objective QOL in outpatients with schizophrenia. We hypothesized that better subjective QOL would be associated with less severe negative and depressive symptoms, better objective QOL, and greater everyday functioning capacity, and that better objective QOL would be associated with less severe negative and depressive symptoms, better cognitive performance, and greater functional capacity. METHOD: Participants included 88 outpatients with schizophrenia or schizoaffective disorder who completed a comprehensive series of assessments, including measures of positive, negative, and depressive symptoms; performance based functional skills; a neuropsychological battery; and an interview measure of subjective and objective QOL. RESULTS: In the context of multiple predictor variables, more severe depressive symptoms and better neuropsychological functioning were independent predictors of worse subjective QOL. More severe negative symptoms predicted worse objective QOL. Functional capacity variables were not associated with subjective or objective QOL. CONCLUSION: Treatments to improve QOL in schizophrenia should focus on negative symptoms and depressive symptoms. PMID- 17919893 TI - VHL protein alterations in sporadic renal cell carcinoma. AB - AIMS: The vhl gene is a tumour suppressor gene implicated in renal tumorigenesis in both familial and sporadic renal cell carcinoma (RCC). Alterations in the gene may modify its suppressor function and allow the formation of renal tumours. The purpose of this study was to determine the existence of vhl gene mutations in renal tumour tissue among patients with sporadic RCC and to assess the effects on the structure of the VHL protein. MATERIALS AND METHODS: This was an observational, analytical and descriptive study of 96 patients who had undergone surgery for sporadic RCC. In surgical specimens of tumour tissue, the three exons of the vhl gene were amplified by polymerase chain reaction and subjected to automatic sequencing. The consequences of the mutations detected on the VHL protein were analysed, taking into account the physical and chemical properties of the amino acids changed by the mutations, the location of the alterations in the protein sequence, the degree of conservation throughout evolution, and prediction of the secondary structure of the protein. RESULTS: In total, 22 vhl gene mutations were detected in 21 (21.9%) patients; in particular, 13 exonic point mutations consisting of 11 sense mutations, one silent mutation and one missense mutation, plus five exon deletions and one insertion. The remaining three were intronic mutations. All changes occurred in protein functional domains and in regions that have been well conserved throughout evolution. Two-thirds of the intronic mutations were considered relevant for protein function. Among the mutations detected, 72.7% were considered capable of compromising the VHL protein suppressor function. CONCLUSIONS: Mutations in the vhl gene result in amino acid changes in the protein that usually occur at important functional sites that have been conserved throughout evolution and where the binding domains for other proteins are located and exert their suppressor function. PMID- 17919895 TI - Phosphate fertilizer from sewage sludge ash (SSA). AB - Ashes from sewage sludge incineration are rich in phosphorus content, ranging between 4% and 9%. Due to the current methods of disposal used for these ashes, phosphorus, which is a valuable plant nutrient, is removed from biological cycling. This article proposes the possible three-stage processing of SSA, whereby more than 90% of phosphorus can be extracted to make an adequate phosphate fertilizer. SSA from two Swiss sewage sludge incinerators was used for laboratory investigations. In an initial step, SSA was leached with sulfuric acid using a liquid-to-solid ratio of 2. The leaching time and pH required for high phosphorus dissolution were determined. Inevitably, dissolution of heavy metals takes place that would contaminate the fertilizer. Thus in a second step, leach solution has to be purified by having the heavy metals removed. Both ion exchange using chelating resins and sulfide precipitation turned out to be suitable for removing critical Cu, Ni and Cd. Thirdly, phosphates were precipitated as calcium phosphates with lime water. The resulting phosphate sludge was dewatered, dried and ground to get a powdery fertilizer whose efficacy was demonstrated by plant tests in a greenhouse. By measuring the weight of plants after 6 weeks of growth, fertilized in part with conventional phosphate fertilizer, fertilizer made from SSA was proven to be equal in its plant uptake efficiency. PMID- 17919894 TI - The effect of pioglitazone as add-on therapy to metformin or sulphonylurea compared to a fixed-dose combination of metformin and glibenclamide on diabetic dyslipidaemia. AB - BACKGROUND AND AIMS: Diabetic dyslipidaemia contributes to the increased risk of cardiovascular disease in patients with Type 2 diabetes. This paper examines the effectiveness of adding pioglitazone to metformin or a sulphonylurea (SU) compared with a fixed-dose combination of metformin and glibenclamide on diabetic dyslipidaemia in patients with Type 2 diabetes. METHODS AND RESULTS: Patients (n=250) treated with metformin (< or =3g/day) or an SU as monotherapy at a stable dose for > or =3 months were randomised to receive either pioglitazone (15-30 mg/day) in addition to their metformin or SU, or a fixed-dose combination tablet containing metformin (400mg) and glibenclamide (2.5 mg) [up to 3 tablets daily] for 6 months. Addition of pioglitazone tended to increase plasma high-density lipoprotein-cholesterol (HDL-C) [0.04 mmol/L; P=0.051] at 6 months and significantly reduced plasma triglycerides (-0.25 mmol/L; P=0.013) compared with baseline. Patients treated with metformin/glibenclamide for 6 months had reduced HDL-C (-0.09 mmol/L; P<0.01) and no change in plasma triglyceride levels (0.03 mmol/L; P=0.733). Both treatment regimes resulted in a similar level of glycaemic control. CONCLUSION: The beneficial effects of pioglitazone on diabetic dyslipidaemia may help combat the increased cardiovascular morbidity and mortality observed in patients with Type 2 diabetes while providing stable glycaemic control. PMID- 17919896 TI - Energy recovery from sewage sludge by means of fluidised bed gasification. AB - Because of its potential harmful impact on the environment, disposal of sewage sludge is becoming a major problem all over the world. Today the available disposal measures are at the crossroads. One alternative would be to continue its usage as fertiliser or to abandon it. Due to the discussions about soil contamination caused by sewage sludge, some countries have already prohibited its application in agriculture. In these countries, thermal treatment is now presenting the most common alternative. This report describes two suitable methods to directly convert sewage sludge into useful energy on-site at the wastewater treatment plant. Both processes consist mainly of four devices: dewatering and drying of the sewage sludge, gasification by means of fluidised bed technology (followed by a gas cleaning step) and production of useful energy via CHP units as the final step. The process described first (ETVS-Process) is using a high pressure technique for the initial dewatering and a fluidised bed technology utilising waste heat from the overall process for drying. In the second process (NTVS-Process) in addition to the waste heat, solar radiation is utilised. The subsequent measures--gasification, gas cleaning and electric and thermal power generation--are identical in both processes. The ETVS-Process and the NTVS-Process are self-sustaining in terms of energy use; actually a surplus of heat and electricity is generated in both processes. PMID- 17919897 TI - Evaluating impact level of different factors in environmental impact assessment for incinerator plants using GM (1, N) model. AB - In this study, the impact levels in environmental impact assessment (EIA) reports of 10 incinerator plants were quantified and discussed. The relationship between the quantified impact levels and the plant scale factors of BeiTou, LiZe, BaLi, LuTsao, RenWu, PingTung, SiJhou and HsinChu were constructed, and the impact levels of the GangShan (GS) and YongKong (YK) plants were predicted using grey model GM (1, N). Finally, the effects of plant scale factors on impact levels were evaluated using grey model GM (1, N) too. According to the predicted results of GM, the relative errors of topography/geology/soil, air quality, hydrology/water quality, solid waste, noise, terrestrial fauna/flora, aquatic fauna/flora and traffic in the GS plant were 17%, 14%, 15%, 17%, 75%, 16%, 13%, and 37%, respectively. The relative errors of the same environmental items in the YK plant were 1%, 18%, 10%, 40%, 37%, 3%, 25% and 33%, respectively. According to GM (1, N), design capacity (DC) and heat value (HV) were the plant scale factors that affected the impact levels significantly in each environmental item, and thus were the most significant plant scale factors. GM (1, N) was effective in predicting the environmental impact and analyzing the reasonableness of the impact. If there is an EIA for a new incinerator plant to be reviewed in the future, the official committee of the Taiwan EPA could review the reasonableness of impact levels in EIA reports quickly. PMID- 17919898 TI - Give-and-take: interactions between DNA transposons and their host plant genomes. AB - Recent genome sequencing efforts have revealed how extensively transposable elements (TEs) have contributed to the shaping of present day plant genomes. DNA transposons associate preferentially with the euchromatic or genic component of plant genomes and have had the opportunity to interact intimately with the genes of the plant host. These interactions have resulted in TEs acquiring host sequences, forming chimeric genes through exon shuffling, replacing regulatory sequences, mobilizing genes around the genome, and contributing genes to the host. The close interaction of transposons with genes has also led to the evolution of intricate cellular mechanisms for silencing transposon activity. Transposons have thus become important subjects of study in understanding epigenetic regulation and, in cases where transposons have amplified to high numbers, how to escape that regulation. PMID- 17919900 TI - Stem cell transplantation in Europe: trends and prospects. AB - The aim of the present study was to identify trends in numbers of European patients treated with autologous and allogeneic haematopoietic stem cell transplantation (HSCT) as well as to provide anticipated transplant rates for the upcoming years. The following indications were considered: haematological malignancies (acute leukaemias, myeloproliferative disorders, lymphoproliferative disorders and multiple myeloma), solid tumours and non-malignant diseases. Numbers of patients treated from 1990 to 2004 were extracted from the European Group for Blood and Marrow Transplantation database, extrapolated to 2012 using mathematic models and adjusted to the literature study and expert opinion. In Europe, a 13% raise in HSCT utilisation is to be expected from 2005 to 2010, mostly due to the growing application of reduced-intensity conditioning regimens followed by allogeneic HSCT. Growing transplant rates are likely to exert health expenditure budgets and put pressure on health care providers and health insurers in Europe. Therefore, the rapid expansion would ideally imply a simultaneous increase in HSCT budgets. PMID- 17919899 TI - Taking it step by step: mechanistic insights from structural studies of ubiquitin/ubiquitin-like protein modification pathways. AB - Ubiquitin (Ub) and ubiquitin-like (Ubl) proteins regulate a diverse array of cellular pathways through post-translational attachment to protein substrates. Ub/Ubl-mediated signaling is initiated through E1, E2, and E3-mediated conjugation, transduced by proteins that recognize Ub/Ubl-modified substrates, and terminated by proteases which remove the Ub/Ubl from the substrate. Recent structural studies have elucidated mechanisms pertinent to Ub/Ubl conjugation, recognition, and deconjugation, highlighting essential steps during Ub/Ubl modification that illustrate common and divergent mechanistic themes within this important process. PMID- 17919901 TI - The effect of propionic to acetic acid ratio on anaerobic-aerobic (low dissolved oxygen) biological phosphorus and nitrogen removal. AB - In this paper, three lab-scale sequencing batch reactors (SBR-A, B, and C) operated with anaerobic/aerobic (low dissolved oxygen, 0.15-0.45 mg L(-1)) configuration were long-term cultured, respectively with single acetic acid and propionic/acetic acid of 1/1 and 2/1 (carbon molar ratio), and the comparisons of anaerobic and aerobic transformations of phosphorus and nitrogen among them were made. With the increase of propionic/acetic acid, lower anaerobic phosphorus release and higher phosphorus release to short-chain fatty acids uptake ratio were observed, and less anaerobic and aerobic transformations of glycogen and poly-3-hydroxybutyrate as well as total polyhydroxyalkanoates occurred, but the transformations of poly-3-hydroxyvalerate and poly-3-hydroxy-2-methyvalerate increased. The phosphorus removal efficiency was respectively 81, 94 and 97% in SBR-A, B and C. Almost all ammonium was removed and no significant nitrite was accumulated at different propionic/acetic acid ratios. However, the nitrate accumulation and total nitrogen removal were observed to be affected by propionic/acetic acid ratio. The total nitrogen removal efficiency was 61, 68 and 82%, and the aerobic end nitrate concentration was 8.05, 6.40 and 3.54 mg L(-1) in three SBRs, respectively. All the above studies indicated that the sole acetic acid caused more nitrate accumulation than propionic and acetic acids mixture, and a pertinent increase of wastewater propionic/acetic acid ratio was of benefit to both nitrogen and phosphorus removal in an anaerobic/aerobic (low dissolved oxygen) biological wastewater treatment process. PMID- 17919902 TI - Enhanced production of biomass, pigments and antioxidant capacity of a nutritionally important cyanobacterium Nostochopsis lobatus. AB - A diazotrophic cyanobacterium Nostochopsis lobatus was evaluated for enhanced production of biomass, pigments and antioxidant capacity. N. lobatus showed potentially high antioxidant capacity (46.12 microM AEAC) with significant improvement under immobilized cell cultures (87.05 microM AEAC). When a mixture of P and Fe was supplemented, biomass, pigments, nutritive value and antioxidant capacity increased substantially at pH 7.8. When considered separately, P appeared to be a better supplement than Fe for the production of biomass, chlorophyll and carotenoids. However, for phycocyanin, phycoerythrin, nutritive value and antioxidant capacity, Fe appeared more effective than P. Our study indicates N. lobatus to be a promising bioresource for enhanced production of nutritionally rich biomass, pigments and antioxidants. The study also suggests that P and Fe are potentially effective supplements for scale-up production for commercial application. PMID- 17919903 TI - Synthesis, antibacterial activity, and quantitative structure-activity relationships of new (Z)-2-(nitroimidazolylmethylene)-3(2H)-benzofuranone derivatives. AB - A new series of (Z)-2-(1-methyl-5-nitroimidazole-2-ylmethylene)-3(2H) benzofuranones (11a-p) and (Z)-2-(1-methyl-4-nitroimidazole-5-ylmethylene)-3(2H) benzofuranones (12a-m) were synthesized and assayed for their antibacterial activity against Gram-positive and Gram-negative bacteria. Most of the 5 nitroimidazole analogues (11a-p) showed a remarkable inhibition of a wide spectrum of Gram-positive bacteria (Staphylococcus aureus, Streptococcus epidermidis, MRSA, and Bacillus subtilis) and Gram-negative Klebsiella pneumoniae, whereas 4-nitroimidazole analogues (12a-m) were not effective against selected bacteria. The quantitative structure-activity relationship investigations were applied to find out the correlation between the experimentally evaluated activities with various parameters of the compounds studied. The QSAR models built in this work had reasonable predictive power and could be explained by the observed trends in activities. PMID- 17919904 TI - Synthesis of aromatic compounds containing a 1,1-dialkyl-2-trifluoromethyl group, a bioisostere of the tert-alkyl moiety. AB - 1,1-Dialkyl-2-perfluoroalkyl compounds, which are potential metabolically stable bioisosteres of the tert-alkyl moiety, have been synthesized from the corresponding tertiary alcohols using titanium (IV) chloride-dimethylzinc or trimethylaluminium as the source of the methyl group. The synthetic methods proved to be versatile for synthesizing 1,1-dimethyl-2,2,2-trifluoroethyl compounds and analogs, including compounds containing aromatic and heterocyclic rings. PMID- 17919905 TI - The discovery of 2-anilinothiazolones as 11beta-HSD1 inhibitors. AB - A series of 2-anilinothiazolones were prepared as inhibitors of 11beta hydroxysteroid dehydrogenase type 1 (11beta-HSD1). The most potent compounds contained a 2-chloro or 2-fluoro group on the aniline ring with an isopropyl substituent on the 5-position of the thiazolone ring (compounds 2 and 3, respectively). The binding mode was determined through the X-ray co-crystal structure of the enzyme with compound 3. This compound was also approximately 70 fold selective over 11beta-HSD2 and was orally bioavailable in rat pharmacokinetic studies. However, compound 3 was >580-fold less active in the 11beta-HSD1 cell assay when tested in the presence of 3% human serum albumin. PMID- 17919906 TI - Salicylic acid inhibits pathogen growth in plants through repression of the auxin signaling pathway. AB - The phytohormone auxin regulates almost every aspect of plant development. At the molecular level, auxin induces gene expression through direct physical interaction with the TIR1-like F box proteins, which in turn remove the Aux/IAA family of transcriptional repressors [1-4]. A growing body of evidence indicates that many plant pathogens can either produce auxin themselves or manipulate host auxin biosynthesis to interfere with the host's normal developmental processes [5 11]. In response, plants probably evolved mechanisms to repress auxin signaling during infection as a defense strategy. Plants overaccumulating the defense signal molecule salicylic acid (SA) frequently display morphological phenotypes that are reminiscent of auxin-deficient or auxin-insensitive mutants, indicating that SA might interfere with auxin responses. By using the Affymetrix ATH1 GeneChip for Arabidopsis thaliana, we performed a comprehensive study of the effects of SA on auxin signaling [12]. We found that SA causes global repression of auxin-related genes, including the TIR1 receptor gene, resulting in stabilization of the Aux/IAA repressor proteins and inhibition of auxin responses. We demonstrate that this inhibitory effect on auxin signaling is a part of the SA-mediated disease-resistance mechanism. PMID- 17919907 TI - Drosophila Spd-2 recruits PCM to the sperm centriole, but is dispensable for centriole duplication. AB - In C. elegans, genome-wide screens have identified just five essential centriole duplication factors: SPD-2, ZYG-1, SAS-5, SAS-6, and SAS-4 [1-8]. These proteins are widely believed to comprise a conserved core duplication module [3, 9-14]. In worm embryos, SPD-2 is the most upstream component of this module, and it is also essential for pericentriolar material (PCM) recruitment to the centrioles [1, 4, 15, 16]. Here, we show that Drosophila Spd-2 (DSpd-2) is a component of both the centrioles and the PCM and has a role in recruiting PCM to the centrioles. DSpd-2 appears not, however, to be essential for centriole duplication in somatic cells. Moreover, PCM recruitment in DSpd-2 mutant somatic cells is only partially compromised, and mitosis appears unperturbed. In contrast, DSpd-2 is essential for proper PCM recruitment to the fertilizing sperm centriole, and hence for microtubule nucleation and pronuclear fusion. DSpd-2 therefore appears to have a particularly important role in recruiting PCM to the sperm centriole. We speculate that the SPD-2 family of proteins might only be absolutely essential for the recruitment of centriole duplication factors and PCM to the centriole(s) that enter the egg with the fertilizing sperm. PMID- 17919908 TI - Straight GDP-tubulin protofilaments form in the presence of taxol. AB - Microtubules exist in dynamic equilibrium, growing and shrinking by the addition or loss of tubulin dimers from the ends of protofilaments. The hydrolysis of GTP in beta-tubulin destabilizes the microtubule lattice by increasing the curvature of protofilaments in the microtubule and putting strain on the lattice. The observation that protofilament curvature depends on GTP hydrolysis suggests that microtubule destabilizers and stabilizers work by modulating the curvature of the microtubule lattice itself. Indeed, the microtubule destabilizer MCAK has been shown to increase the curvature of protofilaments during depolymerization. Here, we show that the atomic force microscopy (AFM) of individual tubulin protofilaments provides sufficient resolution to allow the imaging of single protofilaments in their native environment. By using this assay, we confirm previous results for the effects of GTP hydrolysis and MCAK on the conformation of protofilaments. We go on to show that taxol stabilizes microtubules by straightening the GDP protofilament and slowing down the transition of protofilaments from straight to a curved configuration. PMID- 17919910 TI - Sugar receptors in Drosophila. AB - The detection and discrimination of chemical compounds in potential foods are essential sensory processes when animals feed. The fruit fly Drosophila melanogaster employs 68 different gustatory receptors (GRs) for the detection of mostly nonvolatile chemicals that include sugars, a diverse group of toxic compounds present in many inedible plants and spoiled foods, and pheromones [1 6]. With the exception of a trehalose (GR5a) and a caffeine (GR66a) receptor [7 9], the functions of GRs involved in feeding are unknown. Here, we show that the Gr64 genes encode receptors for numerous sugars. We generated a fly strain that contained a deletion for all six Gr64 genes (DeltaGr64) and showed that these flies exhibit no or a significantly diminished proboscis extension reflex (PER) response when stimulated with glucose, maltose, sucrose, and several other sugars. The only considerable response was detected when Gr64 mutant flies were stimulated with fructose. Interestingly, response to trehalose is also abolished in these flies, even though they contain a functional Gr5a gene, which has been previously shown to encode a receptor for this sugar [8, 9]. This observation indicates that two or more Gr genes are necessary for trehalose detection, suggesting that GRs function as multimeric receptor complexes. Finally, we present evidence that some members of the Gr64 gene family are transcribed as a polycistronic mRNA, providing a mechanism for the coexpression of multiple sugar receptors in the same taste neurons. PMID- 17919911 TI - Two betas or not two betas: regulation of asymmetric division by beta-catenin. AB - In various organisms, cells divide asymmetrically to produce distinct daughter cells. In the nematode Caenorhabditis elegans, asymmetric division is controlled by the asymmetric activity of a Wnt signaling pathway (the Wnt/beta-catenin asymmetry pathway). In this process, two specialized beta-catenin homologs have crucial roles in the transmission of Wnt signals to the asymmetric activity of a T-cell factor (TCF)-type transcription factor, POP-1, in the daughter cells. One beta-catenin homolog regulates the distinct nuclear level of POP-1, and the other functions as a coactivator of POP-1. Both beta-catenins localize asymmetrically in the daughter nuclei using different mechanisms. The recent discovery of reiterative nuclear asymmetries of a highly conserved beta-catenin in an annelid suggests that similar molecular mechanisms might regulate asymmetric cell divisions in other organisms. PMID- 17919909 TI - Zebrafish melanophilin facilitates melanosome dispersion by regulating dynein. AB - BACKGROUND: Fish melanocytes aggregate or disperse their melanosomes in response to the level of intracellular cAMP. The role of cAMP is to regulate both melanosome travel along microtubules and their transfer between microtubules and actin. The factors that are downstream of cAMP and that directly modulate the motors responsible for melanosome transport are not known. To identify these factors, we are characterizing melanosome transport mutants in zebrafish. RESULTS: We report that a mutation (allele j120) in the gene encoding zebrafish melanophilin (Mlpha) interferes with melanosome dispersion downstream of cAMP. Based on mouse genetics, the current model of melanophilin function is that melanophilin links myosin V to melanosomes. The residues responsible for this function are conserved in the zebrafish ortholog. However, if linking myosin V to melanosomes was Mlpha's sole function, elevated cAMP would cause mlpha(j120) mutant melanocytes to hyperdisperse their melanosomes. Yet this is not what we observe. Instead, mutant melanocytes disperse their melanosomes much more slowly than normal and less than halfway to the cell margin. This defect is caused by a failure to suppress minus-end (dynein) motility along microtubules, as shown by tracking individual melanosomes. Disrupting the actin cytoskeleton, which causes wild-type melanocytes to hyperdisperse their melanosomes, does not affect dispersion in mutant melanocytes. Therefore, Mlpha regulates dynein independently of its putative linkage to myosin V. CONCLUSIONS: We propose that cAMP-induced melanosome dispersion depends on the actin-independent suppression of dynein by Mlpha and that Mlpha coordinates the early outward movement of melanosomes along microtubules and their later transfer to actin filaments. PMID- 17919912 TI - Considering theories of aggression in an emergency department context. AB - Internationally literature emphasises concern regarding the phenomenon of violence and aggression within the emergency field. This paper emphasises the important role education and training may play in reducing the risk of staff being exposed to violent or aggressive experiences. Furthermore, the paper emphasises, explores and discusses well recognised theories relating to aggression development. These theories can be used to explain both organisational strategies designed to minimise aggression in the emergency department and situational factors contributing to the development of aggressive interactions. PMID- 17919913 TI - Synthesis and pharmacology of 1-deoxy analogs of CP-47,497 and CP-55,940. AB - A series of 1-deoxy analogs of CP-47,497 (8 and 13, n=0-7) and 1-deoxy analogs of CP-55,940 (9, n=0-7) have been synthesized and their affinities for the cannabinoid CB(1) and CB(2) receptors have been determined. Although the majority of these compounds exhibit selectivity for the CB(2) receptor, none have greater than modest affinity for either receptor. The interactions of these 1-deoxy nontraditional cannabinoids with the CB(2) receptor are discussed. PMID- 17919914 TI - Non-ATP competitive glycogen synthase kinase 3beta (GSK-3beta) inhibitors: study of structural requirements for thiadiazolidinone derivatives. AB - The 2,4-disubstituted thiadiazolidinones (TDZD) were described as the first non ATP competitive GSK-3beta inhibitors. New modifications in this heterocyclic ring are here reported to study the influence on the biological activity. The basic skeleton of 1,2,4-thiadiazole and also one of the carbonyl groups are kept, while different modifications are introduced in positions 3 and 5, respectively. The GSK-3beta activity of the new thiadiazole derivatives here synthesized showed IC(50) values for some of the compounds in the micromolar range. Additionally, ATP competition studies have been carried out, showing that as well as the first generation of TDZD, these new compounds act in a non-competitive manner. With this study, additional requirements for the biological activity of the TDZD family have been delineated. PMID- 17919915 TI - Investigation of the role of the base in the synthesis of [18F]FLT. AB - The role of the base in the synthesis of 3'-deoxy-3'-[18F]fluorothymidine, [18F]FLT, via nucleophilic substitution of the nosyl group with [18F]fluoride was investigated. The rate of 18F-incorporation into the molecule dramatically changed as a function of the precursor-to-base ratio. In the presence of excess base, the precursor was consumed by elimination before substitution was complete. When the precursor-to-base ratio was optimal, an overall [18F]FLT yield of 30-40% was achieved even if the precursor amount was as small as 8-13 mg. PMID- 17919916 TI - Investigations to the synthesis of n.c.a. [18F]FClO3 as electrophilic fluorinating agent. AB - An approach to synthesize the electrophilic fluorinating agent no-carrier-added (n.c.a.) [18F]perchloryl fluoride ([18F]FClO3) in superacidic media in the presence of KClO4 or anhydrous perchloric acid starting from [18F]fluoride was demonstrated in this study. However, the radiochemical yields were low (1-6%) and poorly reproducible. Fluorosulphonic acid proved to be an essential intermediate as revealed by non-radioactive experiments. A key problem in the preparation of [18F]FClO3 is the assumed kinetic inhibition due to the unfavourable stoichiometric ratio of the ClO4 moiety to [18F]HSO3F. PMID- 17919917 TI - Factors which abolish hypoglycemic seizures do not increase cerebral glycogen content in vitro. AB - The brain is heavily dependant on glucose for its function and survival. Hypoglycemia can have severe, irreversible consequences, including seizures, coma and death. However, the in vivo content of brain glycogen, the storage form of glucose, is meager and is a function of both neuronal activity and glucose concentration. In the intact in vitro hippocampus isolated from mice aged postnatal days 8-13, we have recently characterized a novel model of hypoglycemic seizures, wherein seizures were abolished by various neuroprotective strategies. We had hypothesized that these strategies might act, in part, by increasing cerebral glycogen content. In the present experiments, it was found that neither decreasing temperature nor increasing glucose concentrations (above 2 mM) significantly increased hippocampal glycogen content. Preparations of isolated frontal neocortex in vitro do not produce hypoglycemic seizures yet it was found they contained significantly lower glycogen content as compared to the isolated intact hippocampus. Further, the application of either TTX, or a cocktail containing APV, CNQX and gabazine, to block synaptic activity, did not increase, but paradoxically decreased, hippocampal glycogen content in the isolated intact hippocampus. Significant decreases in glycogen were noted when neuronal activity was increased via incubation with l-aspartate (500 muM) or low Mg(2+). Lastly, we examined the incidence of hypoglycemic seizures in hippocampi isolated from mice aged 15-19 and 22-24 days, and compared it to the incidence of hypoglycemic seizures of hippocampi isolated from mice aged 8-13 days described previously (Abdelmalik et al., 2007 Neurobiol Dis 26(3):646-660). It was noted that hypoglycemic seizures were generated less frequently, and had less impact on synaptic transmission in hippocmpi from PD 22-24 as compared to hippocampi from mice PD 15-19 or PD 8-13. However, hippocampi from 8- to 13-day-old mice had significantly more glycogen than the other two age groups. The present data suggest that none of the interventions which abolish hypoglycemic seizures increases glycogen content, and that low glycogen content, per se, may not predispose to the generation of hypoglycemic seizures. PMID- 17919918 TI - Dissecting aneurysm of the interventricular septum following percutaneous balloon aortic valvuloplasty. PMID- 17919919 TI - Serum from exercising humans suppresses t-cell cytokine production. AB - Exercise affects t-cell cytokine production. Whether or not these effects are caused by circulating factors associated with physical activity (e.g., inflammatory mediators, acidosis) is unknown. To investigate this, we incubated sera (10%), obtained from 16 young-adults before (PRE) and after (END) 30-min of exercise, with commercially available Jurkat cells, a t-lymphocyte model, that, of course, had never been exposed to an exercise milieu. After 1 and 6h in culture, we measured in the supernatant four cytokines (each known to be altered by exercise and involved in disease pathophysiology): IL-2, TGF-beta1, TNF-alpha, and IL-1ra. Cell proliferation was assessed with proliferating nuclear cell antigen (PNCA). Statistical analysis consisted of a linear mixed model for repeated measurement. There was no effect of exercise on t-cell production of either TGF-beta1 or IL-1ra. In contrast, both IL-2 (p=0.025) and TNF-alpha (p=0.031) production was significantly suppressed in sera from the exercising participants. The suppression of these two cytokines occurred despite the fact that PNCA significantly increased (p=0.0004) in the END serum. In conclusion, exercise alters circulating factors that can, subsequently, influence t-cell cytokine production in vitro. PMID- 17919921 TI - BDNF mRNA splice variants display activity-dependent targeting to distinct hippocampal laminae. AB - Brain-derived neurotrophic factor (BDNF) may exert contrasting effects depending on its different subcellular sites of action (soma, dendrites, axons). These contrasting effects may explain contradictory findings, for example that BDNF may favour or oppose epileptogenesis. We determined the distribution of five BDNF splice variants in the soma and dendrites of rat hippocampal principal neurons, after application of stimuli that prompt BDNF mRNA accumulation in dendrites (epileptogenic seizures). Under basal conditions, no BDNF mRNA splice variant was detectable in dendrites, while specific splice variants were found in dendrites in response to epileptogenic seizures. Three hours after pilocarpine administration, exon VI and exon II splice variants were found in dendrites, while exons I and IV transcripts displayed a strictly somatic localization. Three hours after kainate administration, only exon VI was found in dendrites. These data suggest that the regulated expression of different splice variants may provide a spatial code to ensure the delivery of BDNF to precise destinations in the cell soma or along the dendrites. PMID- 17919920 TI - Inflammatory cytokines patterns in the placenta of pregnancies complicated by HELLP (hemolysis, elevated liver enzyme, and low platelet) syndrome. AB - OBJECTIVE: We investigated the expression pattern and the role of inflammatory cytokines and their receptors in the placentas of pregnancy with HELLP syndrome. STUDY DESIGN: Placentas were collected after cesarean section, 10 from normal pregnancy and 10 from HELLP. The array was performed with GEArray Q Series Human Inflammatory Cytokines & Receptors Gene Array HS-015. The data were confirmed by quantitative real-time PCR. The Student's t test was used for statistical analysis. RESULTS: Macroarray analysis identified 14 cytokines differentially expressed. PCR confirmed that only IL-10, IL-6-receptor, and TGF-beta3 were increased, whilst CCL18, CXCL5, and IL-16 were significantly decreased, in HELLP. CONCLUSION: The regulation of cytokines involved in angiogenesis and adaptive immune responses may be critical for the placental vascular dysfunction. Our data support the hypothesis that HELLP syndrome could be a placental inflammatory response which leads to a systemic and endothelial dysfunction. PMID- 17919922 TI - Projection maps of three members of the KdgM outer membrane protein family. AB - We present the projection structures of the three outer membrane porins KdgM and KdgN from Erwinia chrysanthemi and NanC from Escherichia coli, based on 2D electron crystallography. A wide screening of 2D crystallization conditions yielded tubular crystals of a suitable size and quality to perform high resolution electron microscopy. Data processing of untilted samples allowed us to separate the information of the two crystalline layers and resulted in projection maps to a resolution of up to 7A. All three proteins exhibit a similar putative beta-barrel structure and the three crystal forms have the same symmetry. However, there are differences in the packing arrangements of the monomers as well as the densities of the projections. To interpret these projections, secondary structure prediction was performed using beta-barrel specific prediction algorithms. The predicted transmembrane beta-barrels have a high similarity in the arrangement of the putative beta-strands and the loops, but do not match those of OmpG, a related protein porin whose structure was solved. PMID- 17919924 TI - Conscious and unconscious proportion effects in masked priming. AB - The global neuronal workspace theory [Dehaene, S., & Naccache, L. (2001). Towards a cognitive neuroscience of consciousness: basic evidence and a workspace framework. Cognition, 79, 1-37.] proposes that it is impossible for an unconscious stimulus to be used strategically to enhance task performance (bottom up effect), while a fully consciously perceived stimulus can be used to improve task performance (top-down effect). Two experiments were designed to investigate these hypotheses. In a first experiment we investigated whether a manipulation of the proportion of Arabic/number word targets had a top-down effect. Since the subjects were fully aware of the target, we expected that this manipulation could be used to enhance task performance. In a second experiment we determined whether a manipulation of the proportion of Arabic/number word primes had a bottom-up effect. Since the subjects were unaware of the prime, we hypothesized that they would be unable to use the manipulation to enhance task performance and therefore the manipulation would have no effect. Both hypotheses were largely confirmed. PMID- 17919923 TI - The conformation of double-stranded DNA inside bacteriophages depends on capsid size and shape. AB - The packaging of double-stranded DNA into bacteriophages leads to the arrangement of the genetic material into highly-packed and ordered structures. Although modern experimental techniques reveal the most probable location of DNA inside viral capsids, the individual conformations of DNA are yet to be determined. In the current study we present the results of molecular dynamics simulations of the DNA packaging into several bacteriophages performed within the framework of a coarse-grained model. The final DNA conformations depend on the size and shape of the capsid, as well as the size of the protein portal, if any. In particular, isometric capsids with small or absent portals tend to form concentric spools, whereas the presence of a large portal favors coaxial spooling; slightly and highly elongated capsids result in folded and twisted toroidal conformations, respectively. The results of the simulations also suggest that the predominant factor in defining the global DNA arrangement inside bacteriophages is the minimization of the bending stress upon packaging. PMID- 17919925 TI - Power calculation for group fMRI studies accounting for arbitrary design and temporal autocorrelation. AB - When planning most scientific studies, one of the first steps is to carry out a power analysis to define a design and sample size that will result in a well powered study. There are limited resources for calculating power for group fMRI studies due to the complexity of the model. Previous approaches for group fMRI power calculation simplify the study design and/or the variance structure in order to make the calculation possible. These approaches limit the designs that can be studied and may result in inaccurate power calculations. We introduce a flexible power calculation model that makes fewer simplifying assumptions, leading to a more accurate power analysis that can be used on a wide variety of study designs. Our power calculation model can be used to obtain region of interest (ROI) summaries of the mean parameters and variance parameters, which can be use to increase understanding of the data as well as calculate power for a future study. Our example illustrates that minimizing cost to achieve 80% power is not as simple as finding the smallest sample size capable of achieving 80% power, since smaller sample sizes require each subject to be scanned longer. PMID- 17919926 TI - Statistical mapping of sound-evoked activity in the mouse auditory midbrain using Mn-enhanced MRI. AB - Manganese-enhanced MRI (MEMRI) has been developed to image brain activity in small animals, including normal and genetically modified mice. Here, we report the use of a MEMRI-based statistical parametric mapping method to analyze sound evoked activity in the mouse auditory midbrain, the inferior colliculus (IC). Acoustic stimuli with defined frequency and amplitude components were shown to activate and enhance neuronal ensembles in the IC. These IC activity patterns were analyzed quantitatively using voxel-based statistical comparisons between groups of mice with or without sound stimulation. Repetitive 40-kHz pure tone stimulation significantly enhanced ventral IC regions, which was confirmed in the statistical maps showing active regions whose volumes increased in direct proportion to the amplitude of the sound stimuli (65 dB, 77 dB, and 89 dB peak sound pressure level). The peak values of the activity-dependent MEMRI signal enhancement also increased from 7% to 20% for the sound amplitudes employed. These results demonstrate that MEMRI statistical mapping can be used to analyze both the 3D spatial patterns and the magnitude of activity evoked by sound stimuli carrying different energy. This represents a significant advance in the development of MEMRI for quantitative and unbiased analysis of brain function in the deep brain nuclei of mice. PMID- 17919927 TI - A simple view of the brain through a frequency-specific functional connectivity measure. AB - Here we develop a measure of functional connectivity describing the degree of covariability between a brain region and the rest of the brain. This measure is based on previous formulas for the mutual information (MI) between clusters of regions in the frequency domain. Under the current scenario, the MI can be given as a simple monotonous function of the multiple coherence and it leads to an easy visual representation of connectivity patterns. Computationally efficient formulas, adequate for short time series, are presented and applied to functional magnetic resonance imaging (fMRI) data measured in subjects (N=34) performing a working memory task or being at rest. While resting state coherence in high (0.17 0.25 Hz) and middle (0.08-0.17 Hz) frequency intervals is bilaterally salient in several limbic and temporal areas including the insula, the amygdala, and the primary auditory cortex, low frequencies (<0.08 Hz) have greatest connectivity in frontal structures. Results from the comparison between resting and N-back conditions show enhanced low frequency coherence in many of the areas previously reported in standard fMRI activation studies of working memory, but task related reductions in high frequency connectivity are also found in regions of the default mode network. Finally, potentially confounding effects of head movement and regional volume on MI are identified and addressed. PMID- 17919928 TI - Bayesian decoding of brain images. AB - This paper introduces a multivariate Bayesian (MVB) scheme to decode or recognise brain states from neuroimages. It resolves the ill-posed many-to-one mapping, from voxel values or data features to a target variable, using a parametric empirical or hierarchical Bayesian model. This model is inverted using standard variational techniques, in this case expectation maximisation, to furnish the model evidence and the conditional density of the model's parameters. This allows one to compare different models or hypotheses about the mapping from functional or structural anatomy to perceptual and behavioural consequences (or their deficits). We frame this approach in terms of decoding measured brain states to predict or classify outcomes using the rhetoric established in pattern classification of neuroimaging data. However, the aim of MVB is not to predict (because the outcomes are known) but to enable inference on different models of structure-function mappings; such as distributed and sparse representations. This allows one to optimise the model itself and produce predictions that outperform standard pattern classification approaches, like support vector machines. Technically, the model inversion and inference uses the same empirical Bayesian procedures developed for ill-posed inverse problems (e.g., source reconstruction in EEG). However, the MVB scheme used here extends this approach to include a greedy search for sparse solutions. It reduces the problem to the same form used in Gaussian process modelling, which affords a generic and efficient scheme for model optimisation and evaluating model evidence. We illustrate MVB using simulated and real data, with a special focus on model comparison; where models can differ in the form of the mapping (i.e., neuronal representation) within one region, or in the (combination of) regions per se. PMID- 17919929 TI - Competition between functional brain networks mediates behavioral variability. AB - Increased intraindividual variability (IIV) is a hallmark of disorders of attention. Recent work has linked these disorders to abnormalities in a "default mode" network, comprising brain regions routinely deactivated during goal directed cognitive tasks. Findings from a study of the neural basis of attentional lapses suggest that a competitive relationship between the "task negative" default mode network and regions of a "task-positive" attentional network is a potential locus of dysfunction in individuals with increased IIV. Resting state studies have shown that this competitive relationship is intrinsically represented in the brain, in the form of a negative correlation or antiphase relationship between spontaneous activity occurring in the two networks. We quantified the negative correlation between these two networks in 26 subjects, during active (Eriksen flanker task) and resting state scans. We hypothesized that the strength of the negative correlation is an index of the degree of regulation of activity in the default mode and task-positive networks and would be positively related to consistent behavioral performance. We found that the strength of the correlation between the two networks varies across individuals. These individual differences appear to be behaviorally relevant, as interindividual variation in the strength of the correlation was significantly related to individual differences in response time variability: the stronger the negative correlation (i.e., the closer to 180 degrees antiphase), the less variable the behavioral performance. This relationship was moderately consistent across resting and task conditions, suggesting that the measure indexes moderately stable individual differences in the integrity of functional brain networks. We discuss the implications of these findings for our understanding of the behavioral significance of spontaneous brain activity, in both healthy and clinical populations. PMID- 17919930 TI - Assessment of brain growth in early childhood using deformation-based morphometry. AB - We present methods for the quantitative analysis of brain growth based on the registration of longitudinal MR image data with the use of Jacobian determinant maps to characterise neuroanatomical changes. The individual anatomies, growth maps and tissue classes are also spatially normalised in an 'average space' and aggregated to provide atlases for the population at each timepoint. The average space representation is obtained using the average intersubject transformation within each timepoint. In an exemplar study, this approach is used to assess brain development in 25 infants between 1 and 2 years, and we show consistency in growth estimates between registration and segmentation approaches. PMID- 17919931 TI - Biophysical model for integrating neuronal activity, EEG, fMRI and metabolism. AB - Our goal is to model the coupling between neuronal activity, cerebral metabolic rates of glucose and oxygen consumption, cerebral blood flow (CBF), electroencephalography (EEG) and blood oxygenation level-dependent (BOLD) responses. In order to accomplish this, two previous models are coupled: a metabolic/hemodynamic model (MHM) for a voxel, linking BOLD signals and neuronal activity, and a neural mass model describing the neuronal dynamics within a voxel and its interactions with voxels of the same area (short-range interactions) and other areas (long-range interactions). For coupling both models, we take as the input to the BOLD model, the number of active synapses within the voxel, that is, the average number of synapses that will receive an action potential within the time unit. This is obtained by considering the action potentials transmitted between neuronal populations within the voxel, as well as those arriving from other voxels. Simulations are carried out for testing the integrated model. Results show that realistic evoked potentials (EP) at electrodes on the scalp surface and the corresponding BOLD signals for each voxel are produced by the model. In another simulation, the alpha rhythm was reproduced and reasonable similarities with experimental data were obtained when calculating correlations between BOLD signals and the alpha power curve. The origin of negative BOLD responses and the characteristics of EEG, PET and BOLD signals in Alzheimer's disease were also studied. PMID- 17919932 TI - Preservation of motor programs in paraplegics as demonstrated by attempted and imagined foot movements. AB - Execution and imagination of movement activate distinct neural circuits, partially overlapping in premotor and parietal areas, basal ganglia and cerebellum. Can long-term deafferented/deefferented patients still differentiate attempted from imagined movements? The attempted execution and motor imagery network of foot movements have been investigated in nine chronic complete spinal cord-injured (SCI) patients using fMRI. Thorough behavioral assessment showed that these patients were able to differentiate between attempted execution and motor imagery. Supporting the outcome of the behavioral assessment, fMRI disclosed specific patterns of activation for movement attempt and for motor imagery. Compared with motor execution data of healthy controls, movement attempt in SCI patients revealed reduced primary motor cortex activation at the group level, although activation was found in all single subjects with a high variability. Further comparisons with healthy subjects revealed that during attempt and motor imagery, SCI patients show enhanced activation and recruitment of additional regions in the parietal lobe and cerebellum that are important in sensorimotor integration. These findings reflect central plastic changes due to altered input and output and suggest that SCI patients may require additional cognitive resources to perform these tasks that may be one and the same phenomenon, or two versions of the same phenomenon, with quantitative differences between the two. Nevertheless, the retained integrity of movement attempt and motor imagery networks in SCI patients demonstrates that chronic paraplegics can still dispose of the full motor programs for foot movements and that therefore, attempted and imagined movements should be integrated in rehabilitative strategies. PMID- 17919933 TI - Changes in white matter microstructure during adolescence. AB - Postmortem histological studies have demonstrated that myelination in human brain white matter (WM) continues throughout adolescence and well into adulthood. We used in vivo diffusion-weighted magnetic resonance imaging to test for age related WM changes in 42 adolescents and 20 young adults. Tract-Based Spatial Statistics (TBSS) analysis of the adolescent data identified widespread age related increases in fractional anisotropy (FA) that were most significant in clusters including the body of the corpus callosum and right superior corona radiata. These changes were driven by changes in perpendicular, rather than parallel, diffusivity. These WM clusters were used as seeds for probabilistic tractography, allowing us to identify the regions as belonging to callosal, corticospinal, and prefrontal tracts. We also performed voxel-based morphometry style analysis of conventional T1-weighted images to test for age-related changes in grey matter (GM). We identified a cluster including right middle frontal and precentral gyri that showed an age-related decrease in GM density through adolescence and connected with the tracts showing age-related WM FA increases. The GM density decrease was highly significantly correlated with the WM FA increase in the connected cluster. Age-related changes in FA were much less prominent in the young adult group, but we did find a significant age-related increase in FA in the right superior longitudinal fascicle, suggesting that structural development of this pathway continues into adulthood. Our results suggest that significant microstructural changes in WM continue throughout adolescence and are associated with corresponding age-related changes in cortical GM regions. PMID- 17919934 TI - The self in conflict: the role of executive processes during truthful and deceptive responses about attitudes. AB - This study sought to extend previous results regarding deceptions about specific memories by investigating the role of executive processes in deceptions about evaluative judgments. In addition, given that previous studies of deception have not included valence manipulations, we also wanted to determine whether the goodness/badness aspect of the items would affect the processes used during deception. Thus, we compared behavioral and event-related potential (ERP) activity while participants made truthful and directed lie (i.e., press opposite of the truth) responses about attitude items with which they either strongly agreed or disagreed. Consistent with previous results, deceptive responses required greater cognitive control as indicated by slower RTs, larger medial frontal negativities (MFN) and smaller late positive components than truthful responses. Furthermore, the magnitude of these deception-related effects was dependent on the valence that participants assigned to the items (i.e., agree/disagree). Directed lie responses about attitudes also resulted in greatly reduced pre-response positivities, an indication that participants strategically monitored their responses even in the absence of explicit task demands. Item valence also differentially affected the amplitude of three ERP components in a 650 ms pre-response interval, independently of whether truthful or deceptive responses were made. Analyses using dipole locations based on results from fMRI studies of evaluative judgments and deception indicated a high degree of overlap between the ERP and fMRI results and revealed the possible temporal characteristics of the hemodynamic activations. PMID- 17919935 TI - Atlas-based segmentation of white matter tracts of the human brain using diffusion tensor tractography and comparison with classical dissection. AB - The technique of diffusion tensor tractography is gaining increasing prominence as a non-invasive method for studying the architecture of the white matter pathways in the human brain. Numerous studies have been published that attempt to identify or reconstruct particular pathways of interest. An atlas or map of all the pathways in the white matter would be particularly useful for providing detailed anatomical data that is not available in studies based on conventional MRI data. In this paper we present a method for constructing a white matter atlas to define structures from diffusion tensor tractography by making use of the locations of the anatomical terminations of individual streamlines that pass through white matter. We show how a map of unique seed regions can be used to generate tracts of interest. This approach provides anatomical information that can be rapidly applied to MRI datasets for the clear identification of white matter tracts. We show close correspondence of the tracts generated from the atlas with tracts isolated with classical dissection of post-mortem brain tissue. PMID- 17919936 TI - Cortical representation of saccular vestibular stimulation: VEMPs in fMRI. AB - Short tone bursts trigger a vestibular evoked myogenic potential (VEMP), an inhibitory potential which reflects a component of the vestibulocollic reflex (VCR). These potentials arise as a result of activation of the sacculus and are expressed through the vestibulo-collic reflex (VCR). Up to now, the ascending projections of the sacculus are unknown in humans, only the representation of the semicircular canals or the entire vestibular nerve has been demonstrated. The aim of this study was to determine whether a sacculus stimulus that evoked VEMPs could activate vestibular cortical areas in fMRI. To determine this, we studied the differential effects of unilateral VEMP stimulation in 21 healthy right handers in a clinical 1.5 T scanner while wearing piezo electric headphones. A unilateral VEMP stimulus and two auditory control stimuli were given in randomized order over the stimulated ear. A random effects statistical analysis was done with SPM2 (p<0.05, corrected). After exclusion of the auditory effects, the major findings were as follows: (i) significant activations were located in the multisensory cortical vestibular network within both hemispheres, including the posterior insular cortex, the middle and superior temporal gyri, and the inferior parietal cortex. (ii) The activation pattern was elicited bilaterally with a predominance of the right hemisphere in right-handers. (iii) Saccular vestibular projection was predominantly ipsilateral, whereas (iv) pure acoustic stimuli were processed with a predominance of the respective contralateral and mainly in the left hemisphere. This is the first demonstration by means of fMRI of the cortical representation of the saccular input at cortical level. The activation pattern is similar to that known from the stimulation of the entire vestibular nerve or the horizontal semicircular canal. Our data give evidence of a task-dependent separation of the processing within the vestibular otolith and the auditory systems in the two hemispheres. PMID- 17919937 TI - Tertiary climate change and the diversification of the Amazonian gecko genus Gonatodes (Sphaerodactylidae, Squamata). AB - The genus Gonatodes is a monophyletic group of small-bodied, diurnal geckos distributed across northern South America, Central America, and the Caribbean. We used fragments of three nuclear genes (RAG2, ACM4, and c-mos) and one mitochondrial gene (16S) to estimate phylogenetic relationships among Amazonian species of Gonatodes. We used Penalized Likelihood to estimate timing of diversification in the genus. Most cladogenesis occurred in the Oligocene and early Miocene and coincided with a burst of diversification in other South American animal groups including mollusks, birds, and mammals. The Oligocene and early Miocene were periods dominated by dramatic climate change and Andean orogeny and we suggest that these factors drove the burst of cladogenesis in Gonatodes geckos as well as other taxa. A common pattern in Amazonian taxa is a biogeographic split between the eastern and western Amazon basin. We observed two clades with this spatial distribution, although large differences in timing of divergence between the east-west taxon pairs indicate that these divergences were not the result of a common vicariant event. PMID- 17919938 TI - Mitogenomic analyses of caniform relationships. AB - Extant members of the order Carnivora split into two basal groups, Caniformia (dog-like carnivorans) and Feliformia (cat-like carnivorans). In this study we address phylogenetic relationships within Caniformia applying various methodological approaches to analyses of complete mitochondrial genomes. Pinnipeds are currently well represented with respect to mitogenomic data and here we add seven mt genomes to the non-pinniped caniform collection. The analyses identified a basal caniform divergence between Cynoidea and Arctoidea. Arctoidea split into three primary groups, Ursidae (including the giant panda), Pinnipedia, and a branch, Musteloidea, which encompassed Ailuridae (red panda), Mephitidae (skunks), Procyonidae (raccoons) and Mustelidae (mustelids). The analyses favored a basal arctoid split between Ursidae and a branch containing Pinnipedia and Musteloidea. Within the Musteloidea there was a preference for a basal divergence between Ailuridae and remaining families. Among the latter, the analyses identified a sister group relationship between Mephitidae and a branch that contained Procyonidae and Mustelidae. The mitogenomic distance between the wolf and the dog was shown to be at the same level as that of basal human divergences. The wolf and the dog are commonly considered as separate species in the popular literature. The mitogenomic result is inconsistent with that understanding at the same time as it provides insight into the time of the domestication of the dog relative to basal human mitogenomic divergences. PMID- 17919939 TI - The strong correlation between alkaline phosphatase activity and cell viability. PMID- 17919940 TI - Altered protein synthesis is a trigger for long-term memory formation. AB - There is ongoing debate concerning whether new protein synthesis is necessary for, or even contributes to, memory formation and storage. This review summarizes a contemporary model proposing a role for altered protein synthesis in memory formation and its subsequent stabilization. One defining aspect of the model is that altered protein synthesis serves as a trigger for memory consolidation. Thus, we propose that specific alterations in the pattern of neuronal protein translation serve as an initial event in long-term memory formation. These specific alterations in protein readout result in the formation of a protein complex that then serves as a nidus for subsequent perpetuating reinforcement by a positive feedback mechanism. The model proposes this scenario as a minimal but requisite component for long-term memory formation. Our description specifies three aspects of prevailing scenarios for the role of altered protein synthesis in memory that we feel will help clarify what, precisely, is typically proposed as the role for protein translation in memory formation. First, that a relatively short initial time window exists wherein specific alterations in the pattern of proteins translated (not overall protein synthesis) is involved in initializing the engram. Second, that a self-perpetuating positive feedback mechanism maintains the altered pattern of protein expression (synthesis or recruitment) locally. Third, that other than the formation and subsequent perpetuation of the unique initializing proteins, ongoing constitutive protein synthesis is all that is minimally necessary for formation and maintenance of the engram. We feel that a clear delineation of these three principles will assist in interpreting the available experimental data, and propose that the available data are consistent with a role for protein synthesis in memory. PMID- 17919941 TI - Interleukin-22 but not interleukin-17 provides protection to hepatocytes during acute liver inflammation. AB - The cytokine interleukin-22 (IL-22) is primarily expressed by T helper 17 (Th17) CD4(+) T cells and is highly upregulated during chronic inflammatory diseases. IL 22 receptor expression is absent on immune cells, but is instead restricted to the tissues, providing signaling directionality from the immune system to the tissues. However, the role of IL-22 in inflammatory responses has been confounded by data suggesting both pro- and anti-inflammatory functions. Herein, we provide evidence that during inflammation, IL-22 played a protective role in preventing tissue injury. Hepatocytes from mice deficient in IL-22 were highly sensitive to the detrimental immune response associated with hepatitis. Additionally, IL-22 expressing Th17 cells provided protection during hepatitis in IL-22-deficient mice. On the other hand, interleukin-17 (IL-17), which is coexpressed with IL-22 and can induce similar cellular responses, had no observable role in liver inflammation. Our data suggest that IL-22 serves as a protective molecule to counteract the destructive nature of the immune response to limit tissue damage. PMID- 17919942 TI - Stimulation of the intracellular bacterial sensor NOD2 programs dendritic cells to promote interleukin-17 production in human memory T cells. AB - How the development of antibacterial T helper 17 (Th17) cells is selectively promoted by antigen-presenting dendritic cells (DCs) is unclear. We showed that bacteria, but not viruses, primed human DCs to promote IL-17 production in memory Th cells through the nucleotide oligomerization domain 2 (NOD2)-ligand muramyldipeptide (MDP), a derivative of bacterial peptidoglycan. MDP enhanced obligate bacterial Toll-like receptor (TLR) agonist induction of IL-23 and IL-1, which promoted IL-17 expression in T cells. The role of NOD2 in this IL-23-IL-1 IL-17 axis could be confirmed in NOD2-deficient DCs, such as DCs from selected Crohn's disease patients. Thus, antibacterial Th17-mediated immunity in humans is orchestrated by DCs upon sensing bacterial NOD2-ligand MDP. PMID- 17919944 TI - Letter to editor re: Dacron or PTFE for above-knee femoropopliteal bypass. A multicenter randomised study. PMID- 17919943 TI - Granzyme B and perforin are important for regulatory T cell-mediated suppression of tumor clearance. AB - Granzyme B is important for the ability of NK cells and CD8(+) T cells to kill their targets. However, we showed here that granzyme B-deficient mice clear both allogeneic and syngeneic tumor cell lines more efficiently than do wild-type (WT) mice. To determine whether regulatory T (Treg) cells utilize granzyme B to suppress immune responses against these tumors, we examined the expression and function of granzyme B in Treg cells. Granzyme B was not expressed in naive Treg cells but was highly expressed in 5%-30% of CD4(+)Foxp3(+) Treg cells in the tumor environment. Adoptive transfer of WT Treg cells, but not granzyme B- or perforin-deficient Treg cells, into granzyme B-deficient mice partially restored susceptibility to tumor growth; Treg cells derived from the tumor environment could induce NK and CD8(+) T cell death in a granzyme B- and perforin-dependent fashion. Granzyme B and perforin are therefore relevant for Treg cell-mediated suppression of tumor clearance in vivo. PMID- 17919945 TI - Near-infrared spectroscopy for evaluation of peripheral vascular disease. A systematic review of literature. AB - OBJECTIVES: To assess near-infrared spectroscopy (NIRS) as a method for the diagnosis and evaluation of peripheral vascular disease. SEARCH STRATEGY: MEDLINE and CENTRAL were searched with a search protocol presented below. Handsearching through reference lists of the retrieved articles and reviews was conducted. MAIN RESULTS: 224 and 57 abstracts from MEDLINE and CENTRAL respectively were retrieved from which 21 studies were selected. NIRS was evaluated for the diagnosis and severity evaluation in patients with peripheral vascular disease. Its parameters were shown to reflect the clinical status of patients, with good correlation to existing methods. CONCLUSIONS: Currently NIRS technology can serve as an adjunct method for the diagnosis and evaluation of patients with peripheral vascular disease. PMID- 17919946 TI - In vitro effects of detergent sclerosants on coagulation, platelets and microparticles. AB - OBJECTIVES: To investigate the in vitro effects of Sodium Tetradecyl Sulphate (STS) and Polidocanol (POL) on clotting tests, clotting factors, platelets and microparticles. MATERIALS AND METHODS: Platelet rich (PRP) and platelet poor (PPP) plasmas were incubated with varying concentrations of STS and POL. Clotting tests, platelet/plasma turbidity, and microparticle studies were performed. Specimens were mixed with individual factor deficient plasmas and clotting factor activities were studied. RESULTS: STS at high concentrations (>0.3%) destroyed platelets, microparticles and the clotting factors V, VII and X. It prolonged all clotting tests including prothrombin time (PT), activated partial thromboplastin time (APTT), non-activated partial thromboplastin time (NAPTT), thrombin time (TT), factor Xa clotting time (XACT) and surface activated clotting time (SACT). Higher concentrations of POL were required to achieve some anticoagulant activity. Low sclerosant concentrations shortened XACT and SACT, and induced release of procoagulant platelet derived microparticles. Increased exposure time resulted in increased procoagulant activity. STS at concentrations higher than 0.5% precipitated a complex containing apolipoprotein b and fibrinogen. CONCLUSIONS: Detergent sclerosants affect the clotting mechanism by interfering with clotting factor activities, procoagulant phospholipids and platelet derived microparticles. STS has more anticoagulant activity than POL in high concentrations. Low concentration sclerosants demonstrate procoagulant activity. PMID- 17919947 TI - Prospective study of the functional recovery after surgery for thoracic outlet syndrome. AB - OBJECTIVES: The aim of this study is to evaluate the functional recovery after Thoracic Outlet Syndrome (TOS) surgery, by the application of Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire. MATERIAL AND METHODS: This was a prospective study of all patients operated on for TOS from January 1998 to December 2005. The DASH questionnaire was administered pre- and postoperatively. The scores were analysed according to TOS type, the associated comorbidity and the type of surgery performed. Results were assessed with Wilcoxon Test for continuous variables, and the Fisher Test for categories. RESULTS: Twenty-three consecutive patients were included in the study, the average age was 37 years (range: 22-54). Fourteen patients presented with venous TOS and 9 with neurogenic TOS. Patients with venous TOS had a preoperative score of 14.9 (SD 18.31) and a postoperative score of 14.8 (SD 15.6) (p>0.05). The preoperative score in patients with neurogenic TOS was 53.96 (SD 15.6) and the postoperative score was 17.8 (SD 15.3) (p=0.01). CONCLUSIONS: DASH questionnaire is a valid and objective test for evaluating the functional state after TOS surgery. Venous TOS is clinically less incapacitating than neurogenic. Surgically decompression of thoracic outlet leads to significant benefit in patients with neurogenic TOS. PMID- 17919948 TI - Ubiquitin-mediated proteasomal degradation in normal and malignant hematopoiesis. AB - Understanding the molecular mechanisms controlling normal hematopoietic differentiation is critical to develop new treatments for blood diseases and to manipulate stem cells. Despite the identification of many players in hematopoiesis, the molecular mechanisms controlling hematopoietic differentiation remain poorly understood. Due to a number of recent findings, the targeting of regulators of hematopoiesis to proteasomal degradation might be an important step in control of this developmental program. PMID- 17919950 TI - Epichloe endophytes grow by intercalary hyphal extension in elongating grass leaves. AB - A fundamental hallmark of fungal growth is that vegetative hyphae grow exclusively by extension at the hyphal tip. However, this model of apical growth is incompatible with endophyte colonization of grasses by the symbiotic Neotyphodium and Epichloe species. These fungi are transmitted through host seed, and colonize aerial tissues that develop from infected shoot apical meristems of the seedling and tillers. We present evidence that vegetative hyphae of Neotyphodium and Epichloe species infect grass leaves via a novel mechanism of growth, intercalary division and extension. Hyphae are attached to enlarging host cells, and cumulative growth along the length of the filament enables the fungus to extend at the same rate as the host. This is the first evidence of intercalary growth in fungi and directly challenges the centuries-old model that fungi grow exclusively at hyphal tips. A new model describing the colonization of grasses by clavicipitaceous endophytes is described. PMID- 17919949 TI - The role of insulator elements in large-scale chromatin structure in interphase. AB - Insulator elements can be classified as enhancer-blocking or barrier insulators depending on whether they interfere with enhancer-promoter interactions or act as barriers against the spreading of heterochromatin. The former class may exert its function at least in part by attaching the chromatin fiber to a nuclear substrate such as the nuclear matrix, resulting in the formation of chromatin loops. The latter class functions by recruiting histone-modifying enzymes, although some barrier insulators have also been shown to create chromatin loops. These loops may correspond to functional nuclear domains containing clusters of co-expressed genes. Thus, insulators may determine specific patterns of nuclear organization that are important in establishing specific programs of gene expression during cell differentiation and development. PMID- 17919951 TI - Canine pododermatitis and idiopathic disease. AB - Pododermatitis is a common inflammatory skin disease of dogs. As pedal lesions are reported in many canine dermatoses, a methodical series of diagnostic tests is required to establish the underlying aetiology. However, laboratory/ancillary investigations may prove unrewarding, prompting a diagnosis of idiopathic disease. Various hypotheses have been proposed to explain the pathogenesis of idiopathic pododermatitis including pedal conformation, trauma, immunosuppression, bacterial infection, furunculosis and dermal granuloma formation. Idiopathic pododermatitis accounts for 0.5% of all dermatology referrals to the authors' clinic. A sub-group within this population is characterised histopathologically by epidermal hyperplasia, hyperkeratosis, spongiosis, dermal oedema and perivascular aggregates of lymphocytes and plasma cells. The term lymphocytic-plasmacytic pododermatitis (LPP) has previously been proposed to reflect the histological appearance of such lesions. Affected dogs, although systemically well, characteristically have pruritus, erythema, swelling, pain and alopecia of the feet. Although non-responsive to antimicrobial therapy, antiparasitic agents and elimination diets, these dogs typically respond well to immunomodulatory therapy. PMID- 17919952 TI - Are reports of mechanical dysfunction in chronic oro-facial pain related to somatisation? A population based study. AB - OBJECTIVES: (i) To examine the association between self-reported mechanical factors and chronic oro-facial pain. (ii) To test the hypothesis that this relationship could be explained by: (a) reporting of psychological factors, (b) common association of self-reported mechanical factors with other unexplained syndromes. METHODS: A population based cross-sectional study of 4200 randomly selected adults registered with a General Medical Practice in North West, England. The study examined the association of chronic oro-facial pain with a variety of self-reported mechanical factors: teeth grinding, facial trauma, missing teeth and the feeling that the teeth did not fit together properly. Information was also collected on demographic factors, psychological factors and the reporting of other frequently unexplained syndromes. RESULTS: An adjusted response rate of 72% was achieved. Only two mechanical factors: teeth grinding (odds ratio (OR) 2.0, 95% CI 1.3-3.0) and facial trauma (OR 2.0; 95% CI 1.3-2.9) were independently associated with chronic oro-facial pain after adjusting for psychological factors. However, these factors were also commonly associated with the reporting of other frequently unexplained syndromes: teeth grinding (odds ratio (OR) 1.8, 95% CI 1.5-2.2), facial trauma (OR 2.1; 95% CI 1.7-2.6). CONCLUSIONS: Self-reported mechanical factors associated with chronic oro-facial pain are confounded, in part, by psychological factors and are equally common across other frequently unexplained syndromes. They may represent another feature of somatisation. Therefore the use of extensive invasive therapy such as occlusal adjustments and surgery to change mechanical factors may not be justified in many cases. PMID- 17919953 TI - Protein assignments without peak lists using higher-order spectra. AB - Despite advances in automating the generation and manipulation of peak lists for assigning biomolecules, there are well-known advantages to working directly with spectra: the eye is still superior to computer algorithms when it comes to picking out peak relationships from contour plots in the presence of confounding factors such as noise, overlap, and spectral artifacts. Here, we present constructs called higher-order spectra for identifying, through direct visual examination, many of the same relationships typically identified by searching peak lists, making them another addition to the set of tools (alongside peak picking and automated assignment) that can be used to solve the assignment problem. The technique is useful for searching for correlated peaks in any spectrum type. Application of this technique to novel, complete sequential assignment of two proteins (AhpFn and IC74(84-143)) is demonstrated. The program "burrow-owl" for the generation and display of higher-order spectra is available at (http://sourceforge.net/projects/burrow-owl) or from the authors. PMID- 17919954 TI - Predicting anti-HIV-1 activity of 6-arylbenzonitriles: computational approach using superaugmented eccentric connectivity topochemical indices. AB - Highly discriminating adjacency-cum-distance based topochemical indices termed as superaugmented eccentric connectivity topochemical indices for quantitative structure-activity and structure-property relationships (QSAR/QSPR) have been conceptualized in the present study. These indices were found to exhibit high sensitivity towards the presence and relative position of heteroatom(s), exceptionally high discriminating power and negligible degeneracy for all possible structures of five vertices containing one heteroatom. Utility of these indices was investigated for development of models for prediction of anti-human immunodeficiency virus (HIV)-1 activity using a data set comprising 81 differently substituted 6-arylbenzonitriles. The values of the superaugmented eccentric connectivity topochemical indices of all the analogues comprising the data set were computed using an in-house computer program. The resultant data was analyzed and suitable models were developed after identification of the active ranges. Subsequently, a biological activity was assigned to each analogue using these models which was then compared with the reported anti-HIV-1 activity. The accuracy of prediction was found to be approximately 81% for all the three topochemical models. High sensitivity towards presence and relative position of heteroatom(s), exceptionally high discriminating power amalgamated with low degeneracy offer proposed topochemical indices vast potential for isomer discrimination, similarity/dissimilarity, drug design, quantitative structure activity/structure-property relationships, lead optimization and combinatorial library design. PMID- 17919955 TI - Hibernation and daily torpor in an armadillo, the pichi (Zaedyus pichiy). AB - Hibernation and daily torpor are physiological strategies to cope with energetic challenges that occur in many mammalian and avian taxa, but no reliable information exists about daily torpor or hibernation for any xenarthran. Our objective was to determine whether the pichi (Zaedyus pichiy), a small armadillo (Xenarthra, Dasypodidae) that inhabits arid and semi-arid habitats in central and southern Argentina and Chile, enters shallow daily torpor or prolonged deep hibernation during winter when environmental temperature and food availability are low. We studied body temperature changes during winter in semi-captive pichis by means of temperature dataloggers implanted subcutaneously. All individuals entered hibernation, characterized by torpor events of 75+/-20 h during which the subcutaneous temperature (T(sc)) decreased to 14.6+/-2.1 degrees C. These events were interrupted by periods of euthermia of 44+/-38 h with a T(sc) of 29.1+/-0.7 degrees C. After the hibernation season, daily torpor bouts of 4 to 6 h occurred irregularly, with T(sc) dropping to as low as 24.5 degrees C. We conclude that the pichi is a true hibernator and can enter daily torpor outside of the hibernation season. PMID- 17919956 TI - Catalase from the white-spotted flower chafer, Protaetia brevitarsis: cDNA sequence, expression, and functional characterization. AB - Catalase, which is one of the key enzymes of the cellular antioxidant defense system, prevents free hydroxyl radical formation by breaking down hydrogen peroxide into oxygen and water. Here, we show the cloning and characterization of a catalase gene in a coleopteran insect. This gene was isolated by searching the white-spotted flower chafer Protaetia brevitarsis cDNA library, and the gene itself encodes a protein of 505 amino acids in length, named PbCat. PbCat shows high similarities to the insect catalase genes known to date. The recombinant PbCat, which is expressed as a 56-kDa polypeptide in baculovirus-infected insect Sf9 cells, shows the highest activity at 30 degrees C and pH 7.0. Northern and Western blot analyses revealed the presence of PbCat in all tissues examined, showing its ubiquitous expression. P. brevitarsis larvae in which H(2)O(2) was overloaded, showed a marked up-regulation in PbCat expression. Moreover, P. brevitarsis larvae showed an apparent increase in PbCat expression even after a wounding through injection. These results indicate that PbCat is up-regulated after wounding and oxidative pressure induced by H(2)O(2), reflecting an important role of PbCat in H(2)O(2) scavenging. PMID- 17919957 TI - Differentiation of ambisexual gonads and immunohistochemical localization of P450 cholesterol side-chain cleavage enzyme during gonadal sex differentiation in the protandrous anemonefish, Amphiprion clarkii. AB - To clarify the relationship between steroid hormones and sex differentiation of the protandrous anemonefish Amphiprion clarkii, we histologically examined its gonadal differentiation. From hatching to 30 days post hatching (dph), all of the gonads surveyed were sexually undifferentiated. The gonads of all fish first differentiated into ovaries at 60 dph, and the oocytes gradually developed and increased in number as the ovaries grew up until 183 dph. Some cysts of differentiated spermatogenic germ cells appeared in the ovaries at 214 dph, and ambisexual gonads with both ovarian and testicular tissues formed by 273 dph. Using immunohistochemistry, we then investigated the expression of cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc), during gonadal sex differentiation. P450scc-immunopositive reactions first appeared in sexually undifferentiated gonads at 30 dph. Beginning at 60 dph, the number of strongly positive cells increased throughout the differentiation of the ovaries and continued to increase during the testicular differentiation until 210 dph. Immunopositive cells were observed more frequently in ovarian tissue than in testicular tissue in the ambisexual gonads at 270 dph. These results suggest that endogenous steroid hormones are important for the sex differentiation, including the primary sex differentiation and subsequent testicular differentiation, of the anemonefish. PMID- 17919958 TI - Kytococcus schroeteri, a rare agent of endocarditis. PMID- 17919959 TI - Systemic effects of epidural and intra-articular glucocorticoid injections in diabetic and non-diabetic patients. AB - INTRODUCTION: Whereas the systemic effects of glucocorticoid therapy have been extensively reported, little is known about those of local glucocorticoid injections. The objective of this study was to look for systemic effects of local glucocorticoid injections at two sites in diabetic and non-diabetic patients. METHODS: We studied 29 patients (18 women and 11 men with an age range of 18-86 years). The injection site was the epidural space in 18 patients (4 with and 14 without diabetes) with disk-related sciatica and the shoulder in 11 patients (8 with and 3 without diabetes) with frozen shoulder. Each patient was given three injections of 1.5 ml cortivazol (5.625 mg of cortivazol or about 85 mg prednisone equivalent per injection and about 250 mg prednisone-equivalent in all), at 3-day intervals. Of the 12 patients with diabetes, 2 were on insulin therapy. At baseline and at the post-treatment visits 1, 7, and 21 days after the third injection, the following tests were done: plasma cortisol and ACTH at 8 am, urinary free cortisol excretion in 24 hours, fasting and postprandial blood glucose, serum cholesterol and triglycerides, and serum sodium and potassium. Blood pressure was measured at each visit. RESULTS: Mean systolic blood pressure increased significantly between baseline (123+/-10 mmHg) and the first two post treatment visits (day 1, 127+/-9 mmHg; and day 7, 128+/-10 mmHg) but returned to baseline values by the third post-treatment visit (day 21). Mean postprandial blood glucose was significantly higher at the day 1 post-treatment visit (10.1+/ 5.4 mmol/l) than at baseline (7.5+/-2.9 mmol/l). At the day 7 post-treatment visit, blood glucose remained significantly elevated compared to baseline in the 12 diabetic patients (13.9+/-4.8 mmol/l versus 9.4+/-3.3 mmol/l at baseline). In both the overall population and the various subgroups, plasma cortisol and ACTH and urinary free cortisol were markedly reduced at the day 1 and day 7 post treatment visits, compared to baseline. At the day 21 visit, these variables were diminished in the group given epidural injections, whereas plasma cortisol and ACTH were normal in the group treated intra-articularly. No significant variations were noted for fasting blood glucose or for serum levels of cholesterol, triglycerides, sodium, and potassium. CONCLUSION: The administration of three local cortivazol injections was followed by suppression of the corticotropic axis that persisted beyond 21 days after epidural injection and recovered more rapidly after intra-articular injection. Systolic blood pressure increased transiently. Elevations in postprandial glucose levels lasted longer in diabetic than non-diabetic patients. PMID- 17919960 TI - Long-term soccer practice increases bone mineral content gain in prepubescent boys. AB - INTRODUCTION: Soccer is a highly osteogenic sport in pubescent adolescents and adults, particularly in weight-bearing bones. However, little is known about its effects in children despite the fact that soccer practice usually starts before puberty. The aim of this study was to verify whether soccer was able to increase bone mineral content (BMC) of weight-bearing bones by modifying bone remodelling in relation to the level of training in prepubescent boys compared to controls. METHODS: At baseline we investigated 39 prepubescent soccer players (11.7+/-0.8 years) divided into two groups according to the duration of training (2 and 4h/week) and 13 controls (10.7+/-0.6 years). BMC and bone resorption marker (CTX) were measured, respectively, by dual-energy X-ray absorptiometry (DEXA) and ELISA. Then, measurements were performed twice during a 10-month survey: only 27 boys remaining prepubescent were analysed. RESULTS: At baseline, no BMC difference was found in weight-bearing sites between soccer players and controls. Nevertheless, soccer players BMC gain significantly increased in total hip (+10.7%, P<0.05), lumbar spine (+10.5%, P<0.05) and legs, the increase being more marked in the longest duration training group (4h/weeks), particularly after a summer resting period. Meanwhile, resorption activity decreased. At the same time, cranial BMC was decreased in soccer players (-4.6%, P<0.001). CONCLUSION: BMC is not significantly enhanced in soccer prepubescent boys in comparison with controls. Nevertheless, the annual gain is greater in soccer players than in controls, especially after a rest period. PMID- 17919961 TI - Septic arthritis of the hip with psoas abscess caused by Non-typhi Salmonella infection in an immunocompetent patient. AB - Osteoarticular infections caused by Non-typhi Salmonella are exceptionally encountered. We report a case of a bacteriologically documented hip infection associated with a psoas abscess due to Non-typhi Salmonella. A 64-year-old immunocompetent male was admitted in our department for pain and motion range limitation in the right hip with fever. Non-typhi Salmonella was recovered in joint fluid obtained by needle aspiration. Antimicrobial chemotherapy combined with surgical intervention was necessary for eradication of the infection. Physicians should be aware of this rare manifestation of Non-typhi Salmonella infections in non-debilitated patients. PMID- 17919962 TI - An efficient and practical synthesis of 2-((1H-indol-3 yl)(aryl)methyl)malononitriles under ultrasound irradiation. AB - Synthesis of 2-((1H-indol-3-yl)(aryl)methyl)malononitrile via the Michael addition of indole with various arylmethylenemalononitriles was carried out in good yields using anhydrous zinc chloride as the catalyst under ultrasound irradiation. PMID- 17919963 TI - Relating mode of action to clinical practice: dopaminergic agents in Parkinson's disease. AB - Most treatment advances in PD have been based on restoring dopaminergic input. The development of levodopa was the first breakthrough and, since then, other compounds have been developed. Each antiparkinsonian medication has its own profile of efficacy and adverse effects, and these can largely be explained by their modes of action. As patients receive a number of different compounds, physicians should be aware of the differences of agents and understand how these differences may relate to clinical practice. This article reviews the three main classes of dopaminergic PD therapy (levodopa, monoamine oxidase inhibitors and dopamine agonists). PMID- 17919964 TI - Neighbourhood social capital and common mental disorder: testing the link in a general population sample. AB - General population multilevel studies of social capital and mental health are few in number. This multilevel study examined external measures of neighbourhood social capital and common mental disorders (CMD). Main effects and stress buffering models were tested. Based on data from over 9000 residents in 239 neighbourhoods in England and Scotland, there was no evidence of a main effect of social capital. For people living in deprived circumstances only, associations between neighbourhood social capital and CMD were seen. Elements of bridging social capital (contact amongst local friends) were associated with lower reporting of CMD. Elements of bonding social capital (attachment to neighbourhood) were associated with higher reporting of CMD. Findings provide some support for the hypothesis that social capital may protect against CMD, but indicate that initiatives should be targeted to deprived groups, focus on specific elements of social capital and not neglect the important relationship between personal socioeconomic disadvantage and CMD. PMID- 17919967 TI - Spectroscopic and theoretical studies on cobalt(II) complex of maleonitriledithiolate and 5-nitro-1,10-phenanthroline. AB - The molecular structure, electronic and infrared spectroscopic properties for the title complex Co(mnt)(5-NO2-phen) (mnt2-=maleonitriledithiolate, 5-NO2-phen=5 nitro-1,10-phenanthroline) were studied in this paper. With semi-empirical PM3 and non-empirical density functional theory (DFT) methods, the gaseous molecular geometry of the complex was optimized and corresponding vibrational spectra was obtained. The calculated results of structure and frequency from DFT were more reasonable than those from PM3, and the two methods were both agreed with the experimental values. A complete assignment to the IR spectra of such a complicated molecule has been exhibited. An electronic spectra was calculated by ZINDOS/S method. The results showed that the calculated values agreed with the observed ones. PMID- 17919965 TI - Amino acid transport through the Saccharomyces cerevisiae Gap1 permease is controlled by the Ras/cAMP pathway. AB - The general amino acid permease (Gap1p) of Saccharomyces cerevisiae is a broad range, low affinity permease that imports amino acids in cells growing on poor nitrogen sources. This permease also signals the presence of amino acids through the fermentable growth medium pathway allowing the cell to respond to new sources of nitrogen in the surrounding medium. Yeast with an activated Ras2/cAMP pathway show many phenotypes indicative of altered nitrogen uptake and metabolism; sensitivity to nitrogen starvation, low amino acid pools. We have shown that Gap1p activity is lowered in cells with an activated RAS2(val19) allele or elevated cAMP levels whereas cells with inactive ras2 allele lose ammonia repression of Gap1p-mediated transport. This regulation is through a post transcriptional mechanism; transcription of GAP1 is not affected by cAMP level. A mechanism by which the Ras2/cAMP/PKA pathway controls the ubiquitin-dependent degradation of Gap1p is most consistent with the data. PMID- 17919968 TI - A simple and efficient colorimetric anion receptor for H(2)PO(4)(-). AB - A novel and neutral anion sensor bearing a urea group as binding sites and 2,4-di nitrophenylhydrazine unit as a molecular architecture and a chromophore was synthesized and the visible color changes, the UV-vis and (1)H NMR spectral responses toward anions were assessed. PMID- 17919969 TI - Density functional theory study on the structure and vibrational frequencies of glycylglycine. AB - Eleven possible conformers of glycylglycine have been studied by using the BLYP, B3LYP methods of density functional theory and the HF method at the basis set of 6-311++G**. BLYP (using Becke's and Lee-Yang-Parr's correlation functionals), ab initio Hartree-Fock (HF) and hybrid DFT/HF B3LYP calculations have been carried out to study the structure and vibrational spectra of glycylglycine. Glycylglycine crystal structure has been determined by X-ray diffraction analysis. The title compound has been crystallizes in the orthorhombic space group C1, with Z=4. And the unit cell parameters are: a=8.1184(12)A, b=9.5542(14)A, c=7.8192(11)A and V=577.95(15)A(3). Molecular conformation calculations have got 11 possible conformers. In these possible conformers, the most stable one has been selected. The BLYP/6-311++G** and scaled HF/6-311++G** frequencies correspond well with available experimental assignments of the normal vibrational modes. Comparison of the observed fundamental vibrational frequencies of glycylglycine and calculated results by density functional B3LYP and Hartree Fock (HF) methods indicates that B3LYP is superior to the scaled Hartree-Fock (HF) for molecular vibrational issues. PMID- 17919970 TI - The vibrational group frequency of the N-O* stretching band of nitroxide stable free radicals. AB - The group frequency of the N-O radical stretching vibration has received scant attention in the literature. The few existing treatments of the vibrational spectroscopy of nitroxides are incomplete at best and potentially misleading to workers in the field. To close this gap in the available knowledge, the existing literature on the vibrational spectra of nitroxide stable free radicals is critically reviewed with particular reference to the wavenumber position of the N O* stretching vibration, nu(N-O*). Poor evidentiary bases for the assignment nu(N O) were found in many instances. Ab initio Density Field Theory calculations using a model chemistry of UB3LYP at the 6-311++G(d,p) level were performed to obtain a theoretical band position of nu(N-O) for comparison with the published data. Large discrepancies between the theoretical and experimental values were found for the radical 3-carbamoyl-2,2,5,5-tetramethyl-3-pyrrolin-1-yloxyl, which currently sets the lower limit of the accepted wavenumber range of nu(N-O*), as well as for the nitronyl and iminyl nitroxides. The wavenumber position of nu(N O*) was found to occur in the range 1450-1420cm(-1) for 5-membered cyclic nitroxides and 1395-1340cm(-1) for 6-membered cyclic and acyclic nitroxides. In nitronyl nitroxides, the symmetric stretching vibration occurs in the region 1470cm(-1), but coupling to other modes makes specific band assignments problematic for the nitronyl nitroxide group. PMID- 17919971 TI - Synthesis, characterization of the luminescent lanthanide complexes with (Z)-4-(4 methoxyphenoxy)-4-oxobut-2-enoic acid. AB - (Z)-4-(4-Methoxyphenoxy)-4-oxobut-2-enoic acid and its solid rare earth complexes LnL3.2H2O (Ln=La, Eu, Tb) were synthesized and characterized by means of MS, elemental analysis, FTIR, 13C NMR and TG-DTA. The IR and 13C NMR results show that the carboxylic groups in the complexes coordinated to the rare earth ions in the form of a bidentate ligand, but the ester carboxylic groups have not taken part in the coordination. The luminescence spectra of Eu(III) and Tb(III) complexes in solid state were also studied. The strong luminescence emitting peaks at 616nm for Eu(III) and 547nm for Tb(III) can be observed, which could be attributed to the ligand has an enhanced effect to the luminescence intensity of the Eu and Tb. PMID- 17919972 TI - Single nucleotide polymorphisms and functional analysis of class II transactivator (CIITA) promoter IV in persistent HBV infection. AB - BACKGROUND/AIMS: CIITA plays a pivotal role in immune response, and the outcome of HBV infection is influenced by immune response. The aims of this study were to analyze the effect of CIITA promoter IV polymorphisms on its activity, and their association with persistent HBV infection. METHODS: The polymorphisms in promoter IV (C-1350T and G-944C) were analyzed by tetra-primer amplification refractory mutation system polymerase chain reaction (ARMS-PCR), and four haplotypes were assigned in 1420 HBV infected subjects. The functional analysis on haplotypes of promoter IV was studied using pGL3-basic and pGL3-promoter vectors. RESULTS: There were significantly decreased-TG and increased-CC haplotype frequencies in persistent HBV infected subjects (34.8% and 41.3%), compared with spontaneously recovered subjects (46.5% and 36.2%). There were significantly higher CC/CC and lower TG/TG genotype frequencies in persistent infected subjects (20.3% and 11.1%) than in spontaneously recovered subjects (10.9% and 23.1%). The mean relative luciferase activity of promoter IV were the highest in TG haplotype (0.558+/-0.023), and the lowest in CC haplotype (0.302+/-0.016). CONCLUSIONS: The polymorphisms of CIITA promoter IV are associated with persistent HBV infection. The CC haplotype with the lowest activity of promoter and CC/CC genotype are responsible for persistent HBV infection. PMID- 17919973 TI - Physical and intellectual development in children with asymptomatic congenital cytomegalovirus infection: a longitudinal cohort study in Qinba mountain area, China. AB - BACKGROUND: Although about 90% of congenital cytomegalovirus (CMV) infection is asymptomatic in newborn, some of them could show sequelae later in life. Qinba mountain area is a place with high incidence of mental retardation and a high rate of CMV intrauterine transmission in China. The correlation between asymptomatic congenital CMV infection and developmental outcomes of children in this area remain unclear. OBJECTIVES: To investigate the impact of asymptomatic congenital CMV infection on physical and intellectual development of children during the first 6 years of life in Qinba mountain area. STUDY DESIGN: Longitudinal cohort study. Forty-nine of all the 54 children with asymptomatic congenital CMV infection were followed prospectively in a study for surveying physical growth and intellectual developments. RESULTS: Either in neonatal or in infant period, no significant difference was noted between the asymptomatic congenital CMV infection children and the controls in average weight, height and head circumference (both p>0.05). The intellectual development was disproportion in asymptomatic congenital infected children. Compared with the control group, both global development quotient (DQ) and full-scale intelligence quotient (IQ) scores of asymptomatically infected children were worse (t=2.19, p=0.031; t=2.48, p=0.015), especially on language DQ scores (t=3.25, p=0.002) and verbal IQ scores (t=3.88, p=0.000). However, the incidence rates of mental retardation (DQ/IQ<70) were similar in two groups (chi(2)=1.03, p>0.05). CONCLUSIONS: Although asymptomatic congenital CMV infection did not have significant influence on the neonatal physical development or incidence of mental retardation later in life, it is obviously an important factor correlating with long-time cognitive outcomes, especially on the development of language. It is necessary to survey CMV congenital infection and monitor the early intellectual development of children with asymptomatic congenital CMV infection in this area. PMID- 17919974 TI - Concealed neo-aortic incompetence and new right ventricular outflow tract obstruction after arterial switch operation in a late presenter. AB - We report a 9 year old boy who presented late with cyanosis as a case of dextro transposition of great arteries, intact ventricular septum and left ventricular outflow tract obstruction (D-TGA/IVS/LVOTO). Arterial switch operation (ASO) with resection of sub-neo aortic membrane and repair of mitral valve were done for the naturally trained LV. On the second postoperative day, the newly discovered right ventricular outflow tract obstruction (RVOTO) was relieved and mitral valve replacement (MVR) was done for significant mixed stenotic/regurgitant mitral valve disease, and intraoperative extra-corporeal membrane oxygenation (ECMO) support was instituted for pulmonary dysfunction for 4 days. Failure of extubation warranted further assessment that revealed significant aortic incompetence (AI) during cardiac catheterisation study, which was underestimated by echocardiography. Aortic valve replacement (AVR) was done on the 11th postoperative day and he was then extubated and had uneventful hospital course in spite of two emergency procedures: drainage of sub-dural haematoma and appendectomy. PMID- 17919975 TI - Monitoring receptor signaling by intramolecular FRET. AB - A large variety of techniques has been used to monitor activation of G protein coupled receptors (GPCRs) both in isolated membranes and in intact cells. However, most of these techniques cannot resolve receptor activation and signaling in space and in time. Here, we describe techniques that allow the temporally and spatially resolved monitoring of these processes by optical recording with energy transfer techniques. Fluorescence and bioluminescence resonance energy transfer, FRET and BRET, are based on energy transfer between two closely spaced probes. The exquisite sensitivity of FRET and BRET to the distance of the two probes makes these techniques ideal tools to study either protein-protein interactions (when the two probes are localized on two different proteins) or conformational changes within a given protein (when the two probes are localized on a single protein). Here, we review the latter approach as a tool to study receptor activation and the levels of the second messengers cAMP and cGMP in intact cells. PMID- 17919976 TI - The peripheral-blood transcriptome: new insights into disease and risk assessment. AB - Future personalized medicine strategies for assessing an individual's health require, ideally, a noninvasive system that is capable of integrating numerous interactive factors, including gender, age, genetics, behavior, environment and comorbidities. Several microarray-based methods developed to meet this goal are currently under investigation. However, most rely on tissue biopsies, which are not readily available or accessible. As an alternative, several recent studies have investigated the use of human peripheral blood cells as surrogate biopsy material. Such studies are based on the assumption that molecular profiling of circulating blood might reflect physiological and pathological events occurring in different tissues of the body. This has led to the development of novel methods for identifying and monitoring blood biomarkers to probe an individual's health status. Here, we discuss the rationale and clinical potential of profiling the peripheral-blood transcriptome. PMID- 17919977 TI - 'Pizza, patients and points of view': Involving young people in the design of a post registration module entitled the adolescent with cancer. AB - Health policy and education directives emphasise the need to involve service users and carers in healthcare, from service design and delivery to the education of healthcare professionals. This paper describes a pilot project aimed at eliciting teenagers and young people with cancer views in relation to the content of a continuing professional development module entitled 'adolescents with cancer'. Seven young people participated in the project. The project design was based on the PARTICIPATION - spice it up [Shephard, C., Treseder, P., 2002. PARTICIPATION: spice it up! Dynamix, Swansea] framework, which aims to ensure effective and appropriate involvement of service users and carers. 'Post-it ideas storm' 'diamond ranking' and 'dot voting' were used to elicit the young people's views. The teenagers and young people could clearly articulate the qualities they felt nurses should have when caring for adolescents with cancer. They also identified topics they felt were important to include in the module, with an understanding of, and using, humour being given the highest priority. This was not originally included in the module timetable but as a direct result of consulting with the young people changes were made to reflect their views. PMID- 17919978 TI - Mutations in DNA repair genes are associated with the Haarlem lineage of Mycobacterium tuberculosis independently of their antibiotic resistance. AB - The analysis of the DNA repair genes ogt and ung was carried out in 117 Mycobacterium tuberculosis clinical isolates from Argentina and Colombia in order to explore correlation between mutations in these genes and multi-drug resistance. With the exception of two Beijing family isolates, the rest of the strains harbored either two wild-type or two mutant alleles with identical single nucleotide polymorphisms (SNPs) in each gene (ogt44 and ung501). These ogt44 and ung501 mutations were not associated with multi-drug resistance and occurred simultaneously in circulating Haarlem genotype M. tuberculosis strains. We therefore propose the use of these markers as tools in phylogenetic and epidemiologic studies. PMID- 17919979 TI - The role of hematopoietic stem cell transplantation in Chronic Lymphocytic Leukemia. AB - Chronic Lymphocytic Leukemia is a common and often incurable lymphoproliferative disorder. Purine nucleoside analogues, and more recently monoclonal antibodies, have increased the potential for obtaining complete or even molecular remissions. Despite these advances, disease recurrence and relapse remain the major concern. Hematopoietic stem cell transplantation is now regularly being considered in this patient population. Attempts at autologous transplantation have not shown additional benefit in advanced refractory disease. However, recent reports show that allogeneic transplantation may represent a chance for cure. This benefit must be balanced with the risk of transplant related mortality and graft-versus host-disease. Current studies suggest that the use of non-myeloablative transplantation may provide durable disease control without the toxicity of conventional transplants. This review will focus on the role of hematopoietic stem cell transplantation in CLL. PMID- 17919980 TI - Family support and cardiac rehabilitation: a comparative study of the experiences of South Asian and White-European patients and their carer's living in the United Kingdom. AB - BACKGROUND: Effective lifestyle modification facilitated by cardiac rehabilitation is known to reduce the occurrence of adverse coronary events and mortality. South Asians have poorer outcomes after a myocardial infarction than the general UK population, but little is known about their experiences of family support, cardiac rehabilitation and lifestyle change. AIMS: To explore the nature of family support available to a sample of South Asian and White-European cardiac patients and to highlight similarities and differences between these groups with regard to cardiac rehabilitation and lifestyle modification. METHODS: Using a qualitative approach, semi-structured interviews (in 1 of 6 languages) were conducted by researchers with; 45 South Asian patients and 37 carers and 20 White European patients and 17 carers. Interviews were conducted in a home setting, up to eighteen months after discharge from hospital following myocardial infarction, coronary artery bypass surgery or unstable angina. RESULTS: The main themes that emerged related to the provision of advice and information, family support and burden, dietary change and exercise regimes. CONCLUSIONS: Several cultural and ethnic differences were identified between patients and their families alongside similarities, irrespective of ethnicity. These may represent generic characteristics of recovery after a cardiac event. Health professionals should develop a cultural repertoire to engage with diversity and difference. Not every difficulty a person encounters as they try to access appropriate service delivery can be attributed to ethnic background. By improving services generally, support for South Asian populations can be improved. The challenge is to know when ethnicity makes a difference and mediates a person's relationship with service support and when it does not. PMID- 17919981 TI - The presence of family members during cardiopulmonary resuscitation: European federation of Critical Care Nursing associations, European Society of Paediatric and Neonatal Intensive Care and European Society of Cardiology Council on Cardiovascular Nursing and Allied Professions Joint Position Statement. AB - This paper presents the European federation of Critical Care Nursing associations, the European Society of Paediatric and Neonatal Intensive Care, and the European Society of Cardiology Council on Cardiovascular Nursing and Allied Professions Joint Position Statement on The Presence of Family Members During Cardiopulmonary Resuscitation. PMID- 17919982 TI - Effects of angiotensin II blockade on the development of autoimmune thyroiditis in nonobese diabetic mice. AB - We evaluated the effects of angiotensin II (Ang II) blockers, losartan, an Ang II receptor blocker, and enalapril, an angiotensin converting enzyme inhibitor, on the development of autoimmune thyroiditis in nonobese diabetic (NOD) mice, an animal model of Hashimoto's thyroiditis (HT). Mice were assigned into three groups, untreated, losartan-treated (30 mg/kg/day), and enalapril-treated (10 mg/kg/day) groups. Thyroiditis was induced by iodide ingestion (experiment 1) or mouse thyroglobulin (Tg) immunization (experiment 2). Both procedures effectively induced thyroiditis. While iodide ingestion failed to induce anti-mouse Tg antibody (TgAb) production, Tg immunization resulted in a significant increase in serum TgAb levels. In both experiments, neither losartan nor enalapril interfered with the development of thyroiditis and TgAb production. These results suggest that Ang II may not be associated with the development of autoimmune thyroiditis in NOD mice. Thus, the Ang II blockade may not have therapeutic potential in HT. PMID- 17919984 TI - From the imaging department: a questionnaire study of the use of radiographs in the evaluation of spine complaints. PMID- 17919983 TI - A technique to measure three-dimensional in vivo rotation of fused and adjacent lumbar vertebrae. AB - BACKGROUND CONTEXT: Previous attempts to measure vertebral motion in vivo have been either static measure, imprecise, two-dimensional, or overly invasive to be applied to serial studies. PURPOSE: This study evaluated the efficacy of a unique high-speed biplane X-ray system for tracking lumbar vertebrae in vivo during dynamic motion. Additional goals were to determine parameters for future studies using this tool and to obtain preliminary data on the effects of lumbar fusion on vertebral kinematics. STUDY DESIGN/SETTING: A high-speed biplane radiographic X ray system was used to measure the three-dimensional (3D) relative rotation between fused and adjacent vertebrae in vivo during muscle driven movement. Subjects were tested 2, 3, and 6 months after fusion procedures to assess vertebral motion of fused and adjacent vertebrae. PATIENT SAMPLE: Five subjects received lumbar fusion surgery. OUTCOME MEASURES: Physiologic measures included 3D vertebral rotation of fused and adjacent vertebrae. METHODS: Tantalum beads were implanted into lumbar vertebrae during fusion operations. Radiographic data was collected continuously at 50 frames per second during flexion-extension, lateral bending, and axial twist movements serially, at 2, 3, and 6 months after fusion surgery. RESULTS: Implanted beads were tracked with an accuracy of 0.18 mm during dynamic motion. Vertebral rotation was not necessarily linearly related to trunk rotation, supporting the use of continuous data collection during movement; collecting only movement start and end points may not be sufficient. Some movements indicated fusion was complete, whereas others indicated incomplete fusion. This suggests patients be tested performing a variety of movements to test for complete fusion. The fusion site often acted as a pivot point for vertebral rotation, with vertebrae superior to the fusion rotating in the direction of the trunk and vertebrae inferior rotating opposite trunk rotation. CONCLUSIONS: This technique is sufficiently accurate for in vivo serial studies of vertebral motion during muscle driven movements. A variety of movements should be performed to assess surgical results, and the data should be collected continuously through the entire range of motion, not just at the movement endpoints. However, care must be exercised in subject selection, in camera location, and in the placement of tracking beads in relation to implanted instrumentation. PMID- 17919985 TI - Evaluating public health uses of health information exchange. AB - Health information exchange (HIE) initiatives are in various stages of development across the United States. They aim to bring previously unavailable clinical data from patients' disparate health records, which may be spread over multiple provider and payer networks, to the point of care where clinicians and their patients need it most. The implications of these initiatives on public health are numerous. This article provides general evaluation methods for measuring the impact of HIE on public health in six use cases: (1) mandated reporting of laboratory diagnoses, (2) mandated reporting of physician-based diagnoses, (3) public health investigation, (4) disease-based non-reportable laboratory data, (5) antibiotic-resistant organism surveillance, and (6) population-level quality monitoring. PMID- 17919986 TI - The United Hospital Fund meeting on evaluating health information exchange. AB - Health information exchange (HIE) projects are sweeping the nation, with hopes that they will lead to high quality, efficient care, yet the literature on their measured benefits remains sparse. To the degree that the field adopts a common set of evaluation strategies, duplicate work can be reduced and meta-analysis will be easier. The United Hospital Fund sponsored a meeting to address HIE evaluation. HIE projects are diverse with many kinds of effects. Assessment of the operation of the HIE infrastructure and of usage should be done for all projects. The immediate business case must be demonstrated for the stakeholders. Rigorous evaluation of the effect on quality may only need to be done for a handful of projects, with simpler process studies elsewhere. Unintended consequences should be monitored. A comprehensive study of return on investment requires an assessment of all effects. Program evaluation across several projects may help set future policy. PMID- 17919987 TI - Efficacy of grape seed proanthocyanidins on serum and heart tissue lipids in rats subjected to isoproterenol-induced myocardial injury. AB - The present study was aimed to evaluate the preventive role of grape seed proanthocyanidins (GSPs) on serum and tissue lipid enzymes in isoproterenol (ISO) induced myocardial injury in male Wistar albino rats. GSP was administered orally to rats (150-180 g) in three different doses, by gastric gavage (50, 100 and 150 mg/kg GSP), 6 days a week for 5 weeks. At the end of this period, all the rats, except the normal untreated rats that served as the control group, were administered ISO, 85 mg/kg subcutaneously, for 2 consecutive days to induce myocardial injury. After 48 h, rats (n=6 per group) were anesthetized with anesthetic ether, sacrificed and the levels of biochemical observations of the serum and heart tissues were performed. Biochemical assessment of myocardial injury was done by measuring the activities of serum thiobarbituric acid reactive substances and plasma lactate, which were significantly elevated in the rats administered with ISO. Further, our results suggest that prior administration of GSPs significantly maintained the cholesterol, phospholipids, triglycerides, and free fatty acids levels in serum and heart tissue of the ISO-induced myocardial injury in rats. The experiments conclude that GSPs possess cardioprotective and hypolipidemic effect on the treatment of ISO-induced myocardial injury. PMID- 17919988 TI - New onset colitis. PMID- 17919989 TI - Gastrointestinal symptoms in 342 patients with Fabry disease: prevalence and response to enzyme replacement therapy. AB - BACKGROUND & AIMS: Fabry disease is an X-linked deficiency of alpha-galactosidase A, resulting in lysosomal deposition of globotriaosylceramide in nearly all tissues. The disease frequently causes diarrhea and abdominal pain, which are assumed to arise from malfunction of enteric neurons and which mimic diarrhea predominant irritable bowel syndrome (IBS). There are limited data about the prevalence and nature of gastrointestinal symptoms in patients with Fabry disease and the response to enzyme replacement therapy (ERT) in large cohorts. The aims of this study were to evaluate the nature and prevalence of gastrointestinal symptoms and their impact on health-related quality of life (HRQoL) in patients with Fabry disease and to analyze changes after 12 and 24 months of treatment with agalsidase alfa. METHODS: Information about gastrointestinal symptoms was obtained from regular interviews before and during the time of ERT. Data on HRQoL were collected by using the EQ-5D questionnaire. RESULTS: The overall prevalence of gastrointestinal symptoms was 52%, with abdominal pain and diarrhea being most frequent. Female patients were more frequently affected than male patients, and there was a high prevalence in children (abdominal pain, 49.3%; diarrhea 25.4%). ERT with agalsidase alfa reduced the prevalence of abdominal pain, with a statistically significant decrease in male patients and in children after 12 months of treatment. CONCLUSIONS: The gastrointestinal symptomatology of Fabry disease is very similar to diarrhea-predominant IBS; however, pathophysiologic similarities remain to be elucidated. ERT reduced the prevalence of gastrointestinal symptoms in Fabry disease, particularly in children and male patients. PMID- 17919991 TI - Drug-induced acute pancreatitis: time for a uniform classification system. PMID- 17919990 TI - HLA DQ gene dosage and risk and severity of celiac disease. AB - BACKGROUND & AIMS: Celiac disease (CD) is a chronic inflammatory disorder of the small intestine that is strongly associated with certain HLA molecules encoded by DQA and DQB genes. The aim of this study was to examine the role of DQA and DQB alleles in determining the risk for and the age of onset and severity of CD in an American population. METHODS: High-resolution class 2 HLA genotyping was performed in a population-based sample (n = 84) of southeastern Minnesota residents with CD and a comparable control group (n = 102) to determine the contribution of DQA and DQB alleles to disease risk. Logistic regression modeling was used to examine the relative and absolute risks of CD. RESULTS: Ninety-seven percent of CD patients carried both of the HLA alleles, DQA1*05 and DQB1*02. Those who carried a second allele of DQB1*02 were 5 times more likely to have CD than those with just one (95% confidence interval, 1.4-18.1). The carriage of 2 copies of DQB1*02 did not predict either an earlier age of onset or severity of disease. CONCLUSIONS: Both HLA alleles DQA1*05 and DQB1*02 are associated with a greatly increased risk of CD, although the latter has the greater effect. Carrying 2 copies of DQB1*02 was associated with an even greater risk for disease but did not predict an earlier age of onset and diagnosis or disease severity. Assessing the copy number of the DQB1*02 allele might allow for the stratification of disease risk. PMID- 17919992 TI - Pharmaceutical company funding and its consequences: a qualitative systematic review. AB - This article systematically reviews published studies of the association of pharmaceutical industry funding and clinical trial results, as well a few closely related studies. It reviews two earlier results, and surveys the recent literature. Results are clear: Pharmaceutical company sponsorship is strongly associated with results that favor the sponsors' interests. PMID- 17919993 TI - Effectiveness of recruitment in clinical trials: an analysis of methods used in a trial for irritable bowel syndrome patients. AB - A successful clinical trial is dependent on recruitment. Between December 2003 and February 2006, our team successfully enrolled 289 participants in a large, single-center, randomized placebo-controlled trial (RCT) studying the impact of the patient-doctor relationship and acupuncture on irritable bowel syndrome (IBS) patients. This paper reports on the effectiveness of standard recruitment methods such as physician referral, newspaper advertisements, fliers, audio and video media (radio and television commercials) as well as relatively new methods not previously extensively reported on such as internet ads, ads in mass-transit vehicles and movie theater previews. We also report the fraction of cost each method consumed and fraction of recruitment each method generated. Our cost per call from potential participants varied from $3-$103 and cost per enrollment participant varied from $12-$584. Using a novel metric, the efficacy index, we found that physician referrals and flyers were the most effective recruitment method in our trial. Despite some methods being more efficient than others, all methods contributed to the successful recruitment. The iterative use of the efficacy index during a recruitment campaign may be helpful to calibrate and focus on the most effective recruitment methods. PMID- 17919994 TI - Gene expression pattern of Cue110: a member of the uncharacterized UPF0224 gene family preferentially expressed in germ cells. AB - The large number of expressed sequence tags (ESTs) now available in databases has enabled the analysis of gene expression profiles in silico. We searched public databases for uncharacterized transcripts specifically expressed in germ cells, in an attempt to identify genes involved in gametogenesis. We found a transcript that is expressed in unfertilized eggs, ovaries, and testes of the mouse. It has an open reading frame (ORF) encoding a 167-amino acid protein belonging to the UPF0224 (unknown protein family 0224) family. We called the novel gene Cue110. We examined the Pfam database for other members of the UPF0224 family, and found a conserved N-terminal portion among members of various species. To study the cellular localization of the Cue110 transcript and protein, we performed in situ hybridization and immunohistochemical analysis of the adult mouse ovary and testis. In the testis, specific hybridization signals were observed weakly in preleptotene spermatocytes but maximally in late round spermatids. Immunostaining showed that Cue110 protein was present predominantly in the cytoplasm of pachytene spermatocytes and round spermatids. In the ovary, weak hybridization signals were observed in primary oocytes in the primordial, primary, and secondary follicles, but Cue110 protein was not detected in oocytes by immunostaining. We next examined the developmental expression pattern of the Cue110 gene using RT-PCR and western blotting, and found its increasing expression coincided with the appearance of spermatocytes. Thus, the Cue110 gene is expressed predominantly in male germ cells at stages from the pachytene spermatocytes to round spermatids. PMID- 17919995 TI - DNA double-strand break and chromosomal rejoining defects with misrejoining in Nijmegen breakage syndrome cells. AB - NBS1-deficient cells exhibit pronounced radiosensitivity and defects in chromosome integrity after ionizing radiation (IR) exposure, yet show only a minor defect in DNA double-strand break (DSB) rejoining, leaving an as yet unresolved enigma as to the nature of the radiosensitivity of these cells. To further investigate the relationship between radiosensitivity, DSB repair, and chromosome stability, we have compared cytological and molecular assays of DSB misrejoining and repair in NBS1-defective, wild type, and NBS1-complemented cells after IR damage. Our findings suggest a subtle defect in overall DSB rejoining in NBS1-defective cells and uniquely also reveal reduced ability of NBS1-defective cells to rejoin correct ends of DSBs. In agreement with published results, one of two different NBS1-defective cell lines showed a slight defect in overall rejoining of DSBs compared to its complemented counterpart, whereas another NBS line did not show any difference from wild type cells. Significant defects in the correct rejoining of DSBs compared to their respective controls were observed for both NBS1-defective lines. The defect in DSB rejoining and the increased misrejoining detected at the molecular level were also reflected in higher levels of fragments and translocations, respectively, at the chromosomal level. This work provides both molecular and cytological evidence that NBS1-deficient cells have defects in DSB processing and reveals that these molecular events can be manifest cytologically. PMID- 17919996 TI - Effects of different nutritional support on lung mechanics and remodelling in undernourished rats. AB - This study investigated the impact of three different oral nutritional support regimens on lung mechanics and remodelling in young undernourished Wistar rats. In the nutritionally deprived group, rats received one-third of their usual daily food consumption for 4 weeks. Undernourished rats were divided into three groups receiving a balanced, glutamine-supplemented, or long-chain triglyceride supplemented diet for 4 weeks. In the two control groups, rats received food ad libitum for 4 (C4) or 8 weeks. Lung viscoelastic pressure and static elastance were higher in undernourished compared to C4 rats. After refeeding, lung mechanical data remained altered except for the glutamine-supplemented group. Undernutrition led to a reduced amount of elastic and collagen fibres in the alveolar septa. Elastic fibre content returned to control with balanced and glutamine-supplemented diets, but increased with long-chain triglyceride supplemented diet. The amount of collagen fibre augmented independent of nutritional support. In conclusion, glutamine-supplemented diet is better at reducing morphofunctional changes than other diets after 4 weeks of refeeding. PMID- 17919997 TI - An efficient method for the purification and quantification of a galactose specific lectin from vegetative tissues of Dolichos lablab. AB - The affinity purified galactose-specific seed lectin from Dolichos lablab, designated as DLL-II, is a tetrameric protein with an apparent native molecular mass of 120 kDa that is composed of two non-identical subunits of 31 and 29 kDa, respectively, associated non-covalently. The stems and leaves of the D. lablab plant also contain a galactose-specific lectin that cross-reacts with the seed lectin antiserum (antiserum raised against the 31 kDa subunit of DLL-II). Anti lectin antibodies have been purified from this antiserum using a gel containing purified DLL-II lectin. Lectin specific antibodies have been used to develop simple and efficient immuno-affinity matrix, which allowed the purification of the lectin from stems and leaves of the D. lablab. The vegetative lectin (DLL-VL) exhibits similar electrophoretic properties as the seed lectin. Using these antibodies, an ELISA method was developed that allowed quantification of the lectin in the vegetative tissues (stems, leaves and roots) at concentrations of 0.5-50 ng. MS and database analysis of the tryptic peptides of the purified subunits of the DLL-VL suggested the purified protein to be a lectin. PMID- 17919998 TI - Simultaneous quantification of GMP, AMP, cyclic GMP and cyclic AMP by liquid chromatography coupled to tandem mass spectrometry. AB - Phosphodiesterases are drug targets for treating various diseases. Inhibition of these can increase cAMP and cGMP levels, which can affect a variety of physiological responses. Here we report a new method for determining PDE activity by combining high-performance liquid chromatography and tandem mass spectrometry. Characteristic peaks of the substrates, cGMP or cAMP and products, GMP or AMP, were identified in positive-ion electrospray ionization using multiple reaction monitoring. The method can be applied to determine activity of PDE inhibitors. Our results showed that this new method was fast, sensitive and highly reproducible. PMID- 17919999 TI - Simultaneous analysis of dextromethorphan and its three metabolites in human plasma using an improved HPLC method with fluorometric detection. AB - A simple and improved HPLC method with fluorometric detection for simultaneous determination of dextromethorphan (DM) and its three metabolites (dextrorphan (DX), 3-methoxymorphinan (MM), 3-hydroxymorphinan (HM)) in human plasma was developed and validated. The method involved a simple and efficient extraction protocol using an n-heptane/ethyl acetate (1:1) solvent mixture that achieved recoveries of 70-90% with an insignificant interference from the plasma matrix. The analysis was performed on a phenyl column with isocratic elution, a mobile phase composed of 20% methanol, 30% acetonitrile, and 50% KH2PO4 buffer (10mM, with adding 0.02% of TEA; adjusted with phosphoric acid to pH 3.5), and a run time of only 15 min. Linear calibration curves were constructed in the concentration range of 1-200 nM for DM and its three metabolites. The lower limit of quantitation (LLOQ) in human plasma was 1 nM for each compound. The coefficient of variation and RSE% of the intraday and interday analyses for DM and its three metabolites all complied with USFDA requirements. This analytical method was preliminarily applied to determine the polymorphic functions of CYP2D6 and CYP3A4 in the metabolic pathway of DM to DX and then to HM. PMID- 17920000 TI - Obesity in Eastern Europe: an overview of its health and economic implications. AB - AIMS: To assess the evolution and patterns of obesity in countries of the WHO European Region with a particular focus on the Eastern European countries, and to discuss the health and economic implications of obesity for those countries. METHODS: The available data on overweight and obesity in children and adults for the countries of the WHO European countries were collated from the International Obesity TaskForce database and considered in the light of estimates for the costs of obesity-related ill health. RESULTS: Overweight and obesity in most countries of Europe show rising secular trends, and are predicted to continue rising if not addressed. Estimates of the costs to the health services and to economic productivity indicate that some countries may find it hard to cope with the burden of obesity: up to 6% of total health care costs and as much in indirect costs of lost productivity could be attributed to obesity and its associated illnesses. CONCLUSIONS: Transition, despite the many benefits it has undoubtedly conferred to the population living in the Region, has also entailed the collateral damage of a fast growing obesity challenge. Policy-makers in the new and candidate EU countries as well as other countries of the European Region can learn from the negative Western European and global experience, act now to stem the obesity epidemic from further developing and in so doing, reduce the substantial economic losses associated with obesity. Local, national and international strategies will be needed to combat the problem. PMID- 17920001 TI - Impacts of Cd(II) on the conformation and self-aggregation of Alzheimer's tau fragment corresponding to the third repeat of microtubule-binding domain. AB - Environmental exposure to some heavy metals such as cadmium appears to be a risk factor for Alzheimer's disease (AD), however, definite mechanism of their toxicity in AD remains to be elucidated. Previous studies largely focused on the metal ions binding to beta-amyloid, however, very few papers concerned the interaction between tau and metal ions. For the first time, we investigated the impacts of Cd(II) on the conformation and self-aggregation of Alzheimer's tau peptide R3, corresponding to the third repeat of microtubule-binding domain. The initial state of R3 was proven to be dimeric linked by intermolecular disulfide bond, in the non-reducing buffer (Tris-HCl buffer pH7.5, containing no reducing reagent). In this paper, we show that Cd(II) can accelerate heparin-induced aggregation of R3 or independently induce the aggregation of R3, as monitored by ThS fluorescence. In the presence of Cd(II), the resulting R3 filaments became much smaller, as revealed by electron microscopy. Binding to the Cd(II) ion, the dimeric R3 partially lost its random coil, and converted to alpha-helix structure, as revealed by CD and Raman spectrum. Stoichiometric analysis of CD signal against the ratio of [Cd(II)]/[R3] suggested that the coordination intermediate consisted of two R3 dimers binding to a central cadmium ion. As the seed, the coordination intermediate could extensively accelerate the self aggregation of R3 via promoting the nucleation step. On the other hand, gain in alpha-helix structure on the peptide chain, by coordinating with Cd(II), could be a critical role to promote self-aggregation, as revealed by Raman spectrum. These results provide a further insight into the mechanism of tau filament formation and emphasize the possible involvement of Cd(II) in the pathogenesis of AD. PMID- 17920002 TI - Promotion of insulin aggregation by protein disulfide isomerase. AB - We examined the aggregation of insulin as a result of reduction of disulfide bonds catalyzed by protein disulfide isomerase (PDI) using various techniques. We demonstrated the kinetic correlation between PDI-catalyzed insulin reduction and the aggregate formation, the relationship between aggregation and amyloid formation, and the structural information on the secondary structure of the aggregates. The initial rate of PDI-catalyzed reduction of insulin, apparent rate constants of aggregate growth for sigmoidal features, and lag times were obtained by changing the PDI concentration, temperature, and insulin concentration. In situ kinetics were studied using the dyes; thioflavin T (ThT) and Congo red (CR) in addition to turbidimetry with the insulin reduction by PDI. The ThT and CR binding assay revealed sigmoidal kinetics, and the spectrum of binding CR showed a red shift against time, suggesting an orderly formation of insulin aggregates. The secondary structure of the PDI-promoted insulin aggregates showed antiparallel beta-sheet conformation by FT-IR measurement. PMID- 17920003 TI - Deregulated expression of Notch receptors in human hepatocellular carcinoma. AB - BACKGROUND AND AIM: Notch signaling controls cellular differentiation and proliferation. Deregulated expression of Notch receptors is observed in a growing number of malignant tumours, however, the role of Notch signaling in hepatocellular carcinoma is still unknown. To address this, the expression of Notch receptors in human hepatocellular carcinoma was examined in both protein and ribonucleic acid levels. PATIENTS AND METHODS: Fifty-three hepatocellular carcinoma tissue sections were detected by immunohistochemistry. Three paired fresh surgical hepatocellular carcinoma and adjacent nontumour liver samples were analyzed by Western blot and reverse transcriptase polymerase chain reaction. Immunohistochemistry, Western blot and reverse transcriptase polymerase chain reaction are reliable methods to examine the expression of protein and RNA. RESULTS: All of the four Notch receptors were expressed in the neoplastic cells of hepatocellular carcinoma tissues with different intensity and extensity. Notch1 and Notch4 were expressed in both cytoplasm and nucleus, and all of the nuclear staining showed up in the cytoplasm-positive cases. Cytoplasmic and nuclear Notch1 was detected in 88.7% (47/53) and 9.4% (5/53) of hepatocellular carcinoma tissues, respectively; positive rates of Notch4 were 67.9% (36/53) in cytoplasm and 52.8% (31/53) in nucleus. Notch2 and Notch3 were only in cytoplasm, with positive rates of 26.4% (14/53) and 52.8% (28/53), respectively. Compared with adjacent nontumour liver, Notch1 (cytoplasmic) and Notch4 (nuclear) were up regulated (P<0.05, P<0.05), Notch2 was down-regulated (P<0.05), while Notch1 (nuclear), Notch3 and Notch4 (cytoplasmic) showed no difference between hepatocellular carcinoma and adjacent nontumour liver. Western blot and reverse transcriptase polymerase chain reaction analysis showed a consistent result. CONCLUSION: Our findings indicate that the expression of Notch receptors was deregulated and Notch signaling might be involved in the development of hepatocellular carcinoma. PMID- 17920005 TI - Efficacy and safety of risedronate 150 mg once a month in the treatment of postmenopausal osteoporosis. AB - INTRODUCTION: Risedronate has been shown to be effective in the treatment of postmenopausal osteoporosis when given orally in daily or weekly doses or on 2 consecutive days per month. This randomized, double-blind, multi-center study was designed to assess the efficacy and safety of a single 150 mg risedronate once-a month oral dose compared with the 5 mg daily regimen. METHODS: Women with postmenopausal osteoporosis were randomly assigned to receive risedronate 5 mg daily (n=642) or 150 mg once a month (followed by daily placebo) (n=650) in a double-blind fashion for 2 years. Study drug was taken on an empty stomach at least 30 min before breakfast. Bone mineral density, bone turnover markers, fractures, and adverse events were evaluated. The primary efficacy endpoint was the mean percent change from baseline in lumbar spine bone mineral density after 1 year. RESULTS: 538 patients in the daily group (83.8%) and 556 patients in the once-a-month group (85.5%) completed 1 year. The mean percent change in lumbar spine bone mineral density was 3.4% (95% confidence interval, 3.03% to 3.82%) in the daily group and 3.5% (95% confidence interval, 3.15% to 3.93%) in the once-a month group. The difference between groups was -0.1% (95% confidence interval, 0.51% to 0.27%). The once-a-month regimen was determined to be non-inferior to the daily regimen based on prospectively defined criteria. The mean percent changes in bone mineral density at sites in the hip (total proximal femur, femoral neck, femoral trochanter) were also similar in both dose groups, as were the changes in biochemical markers of bone turnover. The incidence of adverse events, adverse events leading to withdrawal, and upper gastrointestinal tract adverse events were similar in the 2 treatment groups. Both regimens were well tolerated; the percent of patients who withdrew from treatment as a result of an adverse event was 9.5% in the daily group and 8.6% in the once-a-month group. CONCLUSIONS: Risedronate 150 mg once a month is similar in efficacy and safety to daily dosing and may provide an alternative for patients who prefer once-a-month oral dosing. PMID- 17920004 TI - Surface polyethylene glycol enhances substrate-mediated gene delivery by nonspecifically immobilized complexes. AB - Substrate-mediated gene delivery describes the immobilization of gene therapy vectors to a biomaterial, which enhances gene transfer by exposing adhered cells to elevated DNA concentrations within the local microenvironment. Surface chemistry has been shown to affect transfection by nonspecifically immobilized complexes using self-assembled monolayers (SAMs) of alkanethiols on gold. In this report, SAMs were again used to provide a controlled surface to investigate whether the presence of oligo(ethylene glycol) (EG) groups in a SAM could affect complex morphology and enhance transfection. EG groups were included at percentages that did not affect cell adhesion. Nonspecific complex immobilization to SAMs containing combinations of EG- and carboxylic acid-terminated alkanethiols resulted in substantially greater transfection than surfaces containing no EG groups or SAMs composed of EG groups combined with other functional groups. Enhancement in transfection levels could not be attributed to complex binding densities or release profiles. Atomic force microscopy imaging of immobilized complexes revealed that EG groups within SAMs affected complex size and appearance and could indicate the ability of these surfaces to preserve complex morphology upon binding. The ability to control the morphology of the immobilized complexes and influence transfection levels through surface chemistry could be translated to scaffolds for gene delivery in tissue engineering and diagnostic applications. PMID- 17920006 TI - The ups and downs of neural progenitors: Cep120 and TACCs control interkinetic nuclear migration. AB - The nuclei of dividing neural progenitors undergo a cell-cycle-dependent change in position along the apico-basal axis known as interkinetic nuclear migration (INM). The functional relationship between INM and the mode of division of neural progenitors remains elusive, in part because its regulation at the molecular level is poorly understood. In this issue of Neuron, Xie et al. identify two centrosomal proteins (Cep120 and TACCs) regulating the INM of cortical neural progenitors. PMID- 17920007 TI - Paying attention with the latest technology. AB - In this issue of Neuron, Parikh et al. utilize biosensors to probe the cholinergic system in freely moving rats performing cue-detection/reward delivery tasks. They show that cue-evoked cholinergic activity in the medial prefrontal cortex is associated with cue detection and not reward delivery. We discuss the implications of their research in behavioral neuroscience. PMID- 17920008 TI - The first steps in Drosophila motion detection. AB - The visual system, with its ability to perceive motion, is crucial for most animals to walk or fly steadily. Theoretical models of motion detection exist, but the underlying cellular mechanisms are still poorly understood. In this issue of Neuron, Rister and colleagues dissect the function of neuronal subtypes in the optic lobe of Drosophila to reveal their role in motion detection. PMID- 17920009 TI - The behavioral significance of spontaneous fluctuations in brain activity. AB - Low-frequency fluctuations in fMRI data are believed to reflect synchronous and spontaneous fluctuations in neuronal networks. A study by Fox et al. in this issue of Neuron shows that these spontaneous fluctuations in the motor cortex can account for significant trial-to-trial variations in both the fMRI response and behavior. PMID- 17920010 TI - Self-recognition at the atomic level: understanding the astonishing molecular diversity of homophilic Dscams. AB - The Drosophila Dscams are immunoglobulin superfamily members produced from a single gene that is diversified by alternative splicing to produce a family of cell-surface proteins with over 19,000 different ectodomain isoforms. Dscams are critical for neuronal wiring, and mounting evidence suggests that they play a key role in self-avoidance between sister branches from neurons, which depends on homophilic self-recognition by Dscams. Two recent papers shed new light on Dscam recognition: first by showing that the vast majority of Dscam isoforms mediate specific homophilic binding and second by revealing the essence of the molecular basis of homophilic recognition by Dscams through high-resolution structural studies. PMID- 17920011 TI - To detect and correct: norm violations and their enforcement. AB - Compliance with social norms requires neural signals related both to the norm and to deviations from it. Recent work using economic games between two interacting subjects has uncovered brain responses related to norm compliance and to an individual's strategic outlook during the exchange. These brain responses possess a provocative relationship to those associated with negative emotional outcomes, and hint at computational depictions of emotion processing. PMID- 17920012 TI - Response variation following trauma: a translational neuroscience approach to understanding PTSD. AB - Exposure to traumatic stress is a requirement for the development of posttraumatic stress disorder (PTSD). However, because the majority of trauma exposed persons do not develop PTSD, examination of the typical effects of a stressor will not identify the critical components of PTSD risk or pathogenesis. Rather, PTSD represents a specific phenotype associated with a failure to recover from the normal effects of trauma. Thus, research must focus on identifying pre- and posttraumatic risk factors that explain the development of the disorder and the failure to reinstate physiological homeostasis. In this review, we summarize what is known about the clinical and biological characteristics of PTSD and articulate some of the gaps in knowledge that can be addressed by basic neuroscience research. We emphasize how knowledge about individual differences related to genetic and epigenetic factors in behavioral and brain responses to stress offers the hope of a deeper understanding of PTSD. PMID- 17920013 TI - Perisomatic inhibition. AB - Recent evidence supports the hypothesis of a functional dichotomy of perisomatic inhibition in the cerebral cortex: the parvalbumin- and cholecystokinin containing basket cells that are specialized to control rhythm (as a clockwork) and "mood" (as a fine-tuning device), respectively, of network oscillations. Pathology extends this conclusion further, as the former is implicated in epilepsy and the latter in anxiety. The well-balanced cooperation of the two inhibitory systems is required for the normal network operations underlying the cognitive functions of the cerebral cortex. PMID- 17920014 TI - Imaging large-scale neural activity with cellular resolution in awake, mobile mice. AB - We report a technique for two-photon fluorescence imaging with cellular resolution in awake, behaving mice with minimal motion artifact. The apparatus combines an upright, table-mounted two-photon microscope with a spherical treadmill consisting of a large, air-supported Styrofoam ball. Mice, with implanted cranial windows, are head restrained under the objective while their limbs rest on the ball's upper surface. Following adaptation to head restraint, mice maneuver on the spherical treadmill as their heads remain motionless. Image sequences demonstrate that running-associated brain motion is limited to approximately 2-5 microm. In addition, motion is predominantly in the focal plane, with little out-of-plane motion, making the application of a custom designed Hidden-Markov-Model-based motion correction algorithm useful for postprocessing. Behaviorally correlated calcium transients from large neuronal and astrocytic populations were routinely measured, with an estimated motion induced false positive error rate of <5%. PMID- 17920015 TI - MeCP2 controls excitatory synaptic strength by regulating glutamatergic synapse number. AB - MeCP2 is a transcriptional repressor critical for normal neurological function. Prior studies demonstrated that either loss or doubling of MeCP2 results in postnatal neurodevelopmental disorders. To understand the impact of MeCP2 expression on neuronal function, we studied the synaptic properties of individual neurons from mice that either lack or express twice the normal levels of MeCP2. Hippocampal glutamatergic neurons that lack MeCP2 display a 46% reduction in synaptic response, whereas neurons with doubling of MeCP2 exhibit a 2-fold enhancement in synaptic response. Further analysis shows that these changes were primarily due to the number of synapses formed. These results reveal that MeCP2 is a key rate-limiting factor in regulating glutamatergic synapse formation in early postnatal development and that changes in excitatory synaptic strength may underlie global network alterations in neurological disorders due to altered MeCP2 levels. PMID- 17920017 TI - Cep120 and TACCs control interkinetic nuclear migration and the neural progenitor pool. AB - Centrosome- and microtubule-associated proteins have been shown to be important for maintaining the neural progenitor pool during neocortical development by regulating the mitotic spindle. It remains unclear whether these proteins may control neurogenesis by regulating other microtubule-dependent processes such as nuclear migration. Here, we identify Cep120, a centrosomal protein preferentially expressed in neural progenitors during neocortical development. We demonstrate that silencing Cep120 in the developing neocortex impairs both interkinetic nuclear migration (INM), a characteristic pattern of nuclear movement in neural progenitors, and neural progenitor self-renewal. Furthermore, we show that Cep120 interacts with transforming acidic coiled-coil proteins (TACCs) and that silencing TACCs also causes defects in INM and neural progenitor self-renewal. Our data suggest a critical role for Cep120 and TACCs in both INM and neurogenesis. We propose that sustaining INM may be a mechanism by which microtubule-regulating proteins maintain the neural progenitor pool during neocortical development. PMID- 17920018 TI - Regulation of dendritic development by neuron-specific chromatin remodeling complexes. AB - The diversity of dendritic patterns is one of the fundamental characteristics of neurons and is in part regulated by transcriptional programs initiated by electrical activity. We show that dendritic outgrowth requires a family of combinatorially assembled, neuron-specific chromatin remodeling complexes (nBAF complexes) distinguished by the actin-related protein BAF53b and based on the Brg/Brm ATPases. nBAF complexes bind tightly to the Ca(2+)-responsive dendritic regulator CREST and directly regulate genes essential for dendritic outgrowth. BAF53b is not required for nBAF complex assembly or the interaction with CREST, yet is required for their recruitment to the promoters of specific target genes. The highly homologous BAF53a protein, which is a component of neural progenitor and nonneural BAF complexes, cannot replace BAF53b's role in dendritic development. Remarkably, we find that this functional specificity is conferred by the actin fold subdomain 2 of BAF53b. These studies suggest that the genes encoding the individual subunits of BAF complexes function like letters in a ten letter word to produce biologically specific meanings (in this case dendritic outgrowth) by combinatorial assembly of their products. PMID- 17920016 TI - Amyloid precursor protein regulates brain apolipoprotein E and cholesterol metabolism through lipoprotein receptor LRP1. AB - Mutations in the amyloid precursor protein (APP) cause early-onset Alzheimer's disease (AD), but the only genetic risk factor for late-onset AD is the varepsilon4 allele of apolipoprotein E (apoE), a major cholesterol carrier. Using Cre-lox conditional knockout mice, we demonstrate that lipoprotein receptor LRP1 expression regulates apoE and cholesterol levels within the CNS. We also found that deletion of APP and its homolog APLP2, or components of the gamma-secretase complex, significantly enhanced the expression and function of LRP1, which was reversed by forced expression of the APP intracellular domain (AICD). We further show that AICD, together with Fe65 and Tip60, interacts with the LRP1 promoter and suppresses its transcription. Together, our findings support that the gamma secretase cleavage of APP plays a central role in regulating apoE and cholesterol metabolism in the CNS via LRP1 and establish a biological linkage between APP and apoE, the two major genetic determinants of AD. PMID- 17920019 TI - The BMP ligand Gbb gates the expression of synaptic homeostasis independent of synaptic growth control. AB - Inhibition of postsynaptic glutamate receptors at the Drosophila NMJ initiates a compensatory increase in presynaptic release termed synaptic homeostasis. BMP signaling is necessary for normal synaptic growth and stability. It remains unknown whether BMPs have a specific role during synaptic homeostasis and, if so, whether BMP signaling functions as an instructive retrograde signal that directly modulates presynaptic transmitter release. Here, we demonstrate that the BMP receptor (Wit) and ligand (Gbb) are necessary for the rapid induction of synaptic homeostasis. We also provide evidence that both Wit and Gbb have functions during synaptic homeostasis that are separable from NMJ growth. However, further genetic experiments demonstrate that Gbb does not function as an instructive retrograde signal during synaptic homeostasis. Rather, our data indicate that Wit and Gbb function via the downstream transcription factor Mad and that Mad-mediated signaling is continuously required during development to confer competence of motoneurons to express synaptic homeostasis. PMID- 17920020 TI - Closing in on the resting state of the Shaker K(+) channel. AB - Membrane depolarization causes voltage-gated ion channels to transition from a resting/closed conformation to an activated/open conformation. We used voltage clamp fluorometry to measure protein motion at specific regions of the Shaker Kv channel. This enabled us to construct new structural models of the resting/closed and activated/open states based on the Kv1.2 crystal structure using the Rosetta Membrane method and molecular dynamics simulations. Our models account for the measured gating charge displacement and suggest a molecular mechanism of activation in which the primary voltage sensors, S4s, rotate by approximately 180 degrees as they move "outward" by 6-8 A. A subsequent tilting motion of the S4s and the pore domain helices, S5s, of all four subunits induces a concerted movement of the channel's S4-S5 linkers and S6 helices, allowing ion conduction. Our models are compatible with a wide body of data and resolve apparent contradictions that previously led to several distinct models of voltage sensing. PMID- 17920021 TI - Prefrontal acetylcholine release controls cue detection on multiple timescales. AB - Cholinergic neurons originating from the basal forebrain innervate the entire cortical mantle. Choline-sensitive microelectrodes were used to measure the synaptic release of cortical acetylcholine (ACh) at a subsecond resolution in rats performing a task involving the detection of cues. Cues that were detected, defined behaviorally, evoked transient increases in cholinergic activity (at the scale of seconds) in the medial prefrontal cortex (mPFC), but not in a nonassociational control region (motor cortex). In trials involving missed cues, cholinergic transients were not observed. Cholinergic deafferentation of the mPFC, but not motor cortex, impaired cue detection. Furthermore, decreases and increases in precue cholinergic activity predicted subsequent cue detection or misses, respectively. Finally, cue-evoked cholinergic transients were superimposed over slower (at the timescale of minutes) changes in cholinergic activity. Cortical cholinergic neurotransmission is regulated on multiple timescales to mediate the detection of behaviorally significant cues and to support cognitive performance. PMID- 17920022 TI - Dissection of the peripheral motion channel in the visual system of Drosophila melanogaster. AB - In the eye, visual information is segregated into modalities such as color and motion, these being transferred to the central brain through separate channels. Here, we genetically dissect the achromatic motion channel in the fly Drosophila melanogaster at the level of the first relay station in the brain, the lamina, where it is split into four parallel pathways (L1-L3, amc/T1). The functional relevance of this divergence is little understood. We now show that the two most prominent pathways, L1 and L2, together are necessary and largely sufficient for motion-dependent behavior. At high pattern contrast, the two pathways are redundant. At intermediate contrast, they mediate motion stimuli of opposite polarity, L2 front-to-back, L1 back-to-front motion. At low contrast, L1 and L2 depend upon each other for motion processing. Of the two minor pathways, amc/T1 specifically enhances the L1 pathway at intermediate contrast. L3 appears not to contribute to motion but to orientation behavior. PMID- 17920023 TI - Intrinsic fluctuations within cortical systems account for intertrial variability in human behavior. AB - The resting brain is not silent, but exhibits organized fluctuations in neuronal activity even in the absence of tasks or stimuli. This intrinsic brain activity persists during task performance and contributes to variability in evoked brain responses. What is unknown is if this intrinsic activity also contributes to variability in behavior. In the current fMRI study, we identify a relationship between human brain activity in the left somatomotor cortex and spontaneous trial to-trial variability in button press force. We then demonstrate that 74% of this brain-behavior relationship is attributable to ongoing fluctuations in intrinsic activity similar to those observed during resting fixation. In addition to establishing a functional and behavioral significance of intrinsic brain activity, these results lend new insight into the origins of variability in human behavior. PMID- 17920024 TI - The neural signature of social norm compliance. AB - All known human societies establish social order by punishing violators of social norms. However, little is known about how the brain processes the punishment threat associated with norm violations. We use fMRI to study the neural circuitry behind social norm compliance by comparing a treatment in which norm violations can be punished with a control treatment in which punishment is impossible. Individuals' increase in norm compliance when punishment is possible exhibits a strong positive correlation with activations in the lateral orbitofrontal cortex and right dorsolateral prefrontal cortex. Moreover, lateral orbitofrontal cortex activity is strongly correlated with Machiavellian personality characteristics. These findings indicate a neural network involved in social norm compliance that might constitute an important basis for human sociality. Different activations of this network reveal individual differences in the behavioral response to the punishment threat and might thus provide a deeper understanding of the neurobiological sources of pathologies such as antisocial personality disorder. PMID- 17920025 TI - Identification of a mitochondrial ATP synthase-adenine nucleotide translocator complex in Leishmania. AB - The ATP synthasome is a macromolecular complex consisting of ATP synthase, adenine nucleotide translocator and phosphate carrier. To determine if this complex is evolutionary old or young, we searched for its presence in Leishmania, a mitochondria containing protozoan which evolved from the main eukaryote line soon after eukaryotes split from prokaryotes. Sucrose gradient centrifugation showed that the distribution of ANT among the fractions coincided with the distribution of ATP synthase. In addition, ATP synthase co-precipitated with FLAG tagged and wild type adenine nucleotide translocator isolated with anti FLAG and anti adenine nucleotide translocator antibodies, respectively. These data indicate that the adenine nucleotide translocator interacted with the ATP synthase to form a stable structure referred to as the ATP synthasome. The presence of the ATP synthasome in Leishmania, an organism branching off the main line of eukaryotes early in the development of eukaryotes, as well as in higher eukaryotes suggests that the ATP synthasome is a phylogenetically ancient structure. PMID- 17920026 TI - Artifactual detection of biotin on histones by streptavidin. AB - Biotinylation is a recent addition to the list of reported posttranslational modifications made to histones. Holocarboxylase synthetase (HCS) and biotinidase have been implicated as biotinylating enzymes. However, the details of the mechanism and the regulation of biotin transfer on and off histones remains unclear. Here we report that in a cell culture system low biotin availability reduces biotinylation of carboxylases, yet apparent biotinylation of histones is unaffected. This is despite biotin depletion having detrimental effects on cell viability and proliferation. Further analysis of the widely used method for detecting biotin on histones, streptavidin blotting, revealed that streptavidin interacts with histones independently of biotin binding. Preincubation of streptavidin with free biotin reduced binding to biotinylated carboxylases but did not block binding to histones. To investigate biotinylation of histones using an alternative detection method independent of streptavidin, incorporation of 14C biotin into biotinylated proteins was analyzed. Radiolabeled biotin was readily detectable on carboxylases but not on histones, implying very low levels of biotin in the nucleus attached to histone proteins (< 0.03% biotinylation). In conclusion, we would caution against the use of streptavidin for investigating histone biotinylation. PMID- 17920027 TI - Isothermal titration calorimetry at very low c. AB - In the study of 1:1 binding, M + X right arrow over left arrow MX, isothermal titration calorimetry (ITC) can be used successfully at values of c=K[M](0) well below the value 1.0 that is often considered its lower limit. However, analysis of low-c ITC data may require freezing the stoichiometry parameter n, and that is thought to be prohibitive for biological systems, where n can be poorly known. Here it is noted that the least-squares estimates of the binding constant K are virtually independent of errors in n at low c, permitting reliable determination of K and, from its temperature dependence, DeltaH degrees and n, down to c=10(-4) or lower, ligand solubility permitting. PMID- 17920028 TI - Binding characteristics of bovine serum albumin encapsulated in sol-gel glasses: an alternative for protein interaction studies. AB - Silica glasses doped with 500-700 microg of bovine serum albumin were prepared by the sol-gel method; two pH conditions (pH 5 and 7) were assayed for protein encapsulation. Both biomaterials showed a highly porous structure, with pore sizes in the range 5-28 nm. Columns packed with the ground biogels were on-line coupled to a C18 HPLC column for evaluation of the entrapped protein binding properties using propranolol. Binding capacities (at saturation) were approximately 3.7 and 7.1 microg of propranolol (drug-protein molar ratios 1.4 and 2.7) for the biogels prepared at pH 5 and 7, respectively. The significant difference indicates increased albumin denaturation upon encapsulation at pH 5. A frontal analysis study was then performed in cartridges packed with biogel prepared at pH 7 to evaluate the protein interaction with naproxen at low concentrations (30 days in 2 of 8 patients), the dose was reduced to 10 microg/kg in 24 patients. On day 30, 84% of patients demonstrated a clinical response, 41% achieved clinical remission, and 44% achieved endoscopic remission. Forty-five percent of patients did not require salvage therapies or colectomy during the first year postdose. Mild to moderate symptoms of cytokine release occurred in 100% and 83% of patients in the 15- and 10-microg/kg dose groups, respectively. All patients exhibited a rapid decrease in circulating CD4(+) T-cell counts, which returned to baseline values by day 30 in 26 of 30 evaluable patients (86%). There were no serious infections. CONCLUSIONS: Visilizumab had an acceptable safety profile at the 10-microg/kg dose level and may be clinically beneficial in patients with severe intravenous corticosteroid-refractory ulcerative colitis. PMID- 17920066 TI - The presence of the adult respiratory distress syndrome does not worsen mortality or discharge disability in blunt trauma patients with severe traumatic brain injury. AB - PURPOSE: To evaluate the prevalence of the acute respiratory distress syndrome (ARDS) among blunt trauma patients with severe traumatic brain injury (TBI) and to determine if ARDS is associated with higher mortality, morbidity and worse discharge outcome. METHODS: Blunt trauma patients with TBI (head abbreviated injury score (AIS)> or =4) who developed predefined ARDS criteria between January 2000 and December 2004 were prospectively collected as part of an ongoing ARDS database. Each patient in the TBI+ARDS group was matched with two control TBI patients based on age, injury severity score (ISS) and head AIS. Outcomes including complications, mortality and discharge disability were compared between the two groups. RESULTS: Among 362 TBI patients, 28 (7.7%) developed ARDS. There were no differences between the two groups with respect to age, sex, ISS, Glasgow coma score (GCS), head, abdomen and extremity AIS. The TBI+ARDS group had significantly more patients with chest AIS> or =3 (57.1% versus 32.1%, p=0.03). There was no difference with respect to overall mortality between the TBI+ARDS group (50.0%) and the TBI group (51.8%) (OR 0.79: 95% CI 0.31-2.03, p=0.63). There was no significant difference with respect to discharge functional capacity between the two groups. There were significantly more overall complications in the TBI+ARDS group (42.9%) compared to the TBI group (16.1%) (OR 3.66: 95% CI 1.19-11.24, p=0.02). The TBI+ARDS group had an overall mean intensive care unit (ICU) length of stay of 15.6 days, versus 8.4 days in the TBI group (p<0.01). The TBI+ARDS group had significantly higher hospital charges than the TBI group ($210,097 versus $115,342, p<0.01). CONCLUSION: The presence of ARDS was not associated with higher mortality or worse discharge disability. It was, however, associated with higher hospital morbidity, longer ICU and hospital length of stay. PMID- 17920067 TI - The progress of coronary heart disease in Type 2 diabetes as measured by coronary calcium score from electron beam computed tomography (EBCT): the PREDICT study. AB - Coronary calcification score (CACS) measured by electron beam tomography is well established in the evaluation of cardiovascular risk in general populations. The PREDICT study aims to evaluate prediction of cardiovascular events by CACS in Type 2 diabetic subjects without previous clinical cardiovascular disease. In the present PREDICT sub-study, the rate of progression of CACS and factors influencing this rate were assessed. CACS was measured at baseline and after a mean interval of 4.0 (range of 2.1-5.0) years in the 202 PREDICT participants who agreed to have a second scan. Participants also had a range of conventional and novel biochemical risk factors measured at baseline. Progression of calcification was apparent in 170 (84%), while in 32 (16%) there was regression or no progression. Those showing progression had a significantly more adverse risk factor profile. Rate of change in CACS was strongly related to baseline CACS (p<0.0001). Rate of change also correlated with, waist:hip ratio (p=0.004), male gender (p=0.009), age (p=0.04), use of antihypertensive drugs (p=0.03) and statins (p=0.05) and, independently of baseline CACS, systolic blood pressure (p=0.0006), waist circumference (p=0.001) and urine albumin:creatine ratio (p=0.04). Most subjects with Type 2 diabetes showed progression of CACS. The absence of a relationship between progression and lipid risk factors and the positive relationship with statin and antihypertensive drug use may reflect earlier risk factor exposure. Independent relationships between progression and established calcification, blood pressure, central adiposity and urine albumin:creatinine ratio suggest areas for risk factor modification that could be especially relevant in Type 2 diabetes. PMID- 17920068 TI - Realistic finite element-based stent design: the impact of balloon folding. AB - At present, the deployment of an intravascular stent has become a common and widely used minimally invasive treatment for coronary heart disease. To improve these coronary revascularization procedures (e.g. reduce in-stent restenosis rates) the optimal strategy lies in the further development of stent design, material and coatings. In the context of optimizing the stent design, computational models can provide an excellent research tool. In this study, the hypothesis that the free expansion of a stent is determined by the unfolding and expansion of the balloon is examined. Different expansion modeling strategies are studied and compared for a new generation balloon-expandable coronary stent. The trifolded balloon methodology presented in this paper shows very good qualitative and quantitative agreement with both manufacturer's data and experiments. Therefore, the proposed numerical expansion strategy appears to be a very promising optimization methodology in stent design. PMID- 17920069 TI - Determination of sequestering agents in cosmetics and synthetic detergents by high-performance liquid chromatography with ultraviolet detection. AB - Using a fast reversible reaction of aminopolycarboxylic acids (APCAs) into Fe(III)-APCA complexes in the presence of Fe(III) ions, seven kinds of APCAs [nitrilotriacetate (NTA), N-(2-hydroxyethyl)ethylenediamine-triacetate (HEDTA), ethylenediamine-tetraacetate (EDTA), 1,3-propanediamine-tetraacetate (PDTA), diethylenetriamine-pentaacetate (DTPA), 1,2-diaminopropane-tetraacetate (MeEDTA), and O,O'-bis(2-aminoethyl)ethyleneglycol-tetraacetate (GEDTA)] in cosmetics and synthetic detergents were separated on two reversed-phase C30 columns connected in series and detected with ultraviolet detection. Simple pretreatment, consisted of thousand times dilution of samples and addition of 100 microl of the Fe(III) solution containing 10 mM Fe(III) chloride and 0.5 M sulfuric acid to 10 ml of diluted samples, permitted the determination of APCAs in cosmetics and synthetic detergents at concentration level of 0.1 mM, except 0.3 mM for GEDTA. APCAs except GEDTA could be detected at concentration level of 0.03 mM and GEDTA could be detected at concentration level of 0.09 mM. Good recoveries (95-110%) were obtained for each APCA by the standard addition method on two diluted samples with high accuracy (RSD 0.2-9.1%). Three APCAs (EDTA, HEDTA and NTA) were detected in various concentrations in cosmetics and synthetic detergents and the other APCAs were not detected in any of the samples. This method requires no tedious pretreatment and takes only 15 min for one analysis, so it is useful for determination of APCAs. PMID- 17920070 TI - Post-column reactor of coaxial-gap mode for laser-induced fluorescence detection in capillary electrophoresis. AB - A post-column reactor with coaxial-gap mode is developed for laser-induced fluorescence detection (LIF) in capillary electrophoresis (CE). The reactor can be assembled simply and conveniently, in which a thin polyimide sleeve of 10-mm length obtained from the capillary coating is used to align separation and reaction capillary with a 20 microm gap. Naphthalene-2,3-dicarboxaldehyde and 2 mercaptoethanol are used as derivatization reagents and delivered into the reaction capillary through the annulus between the separation capillary and polyimide sleeve and the gap of two capillaries by gravity. A reaction distance from the gap to detection point is 5mm. For the post-column reactor of CE-LIF, several configuration parameters are optimized, including liquid level difference between the derivatization solution and outlet buffer, annular dimension between the outer diameter of etched separation capillary and the inner diameter of polyimide sleeve, and reaction distance, etc. The detection limits in the range from 8.0x10(-8) to 1.0x10(-6) mol/L and linear calibration range more than two orders of magnitude are obtained for amino acids. The separation efficiency ranges from 1.35x10(5) to 1.67x10(5) theoretical plates. PMID- 17920071 TI - As-placed contact angles for sessile drops. AB - As-placed contact angle is the contact angle a drop adapts as a result of its placement on a surface. As expected, the as-placed contact angle, thetaAP, of a sessile drop on a horizontal surface decreases with the drop size due to the increase in hydrostatic pressure. We present a theoretical prediction for thetaAP which shows that it is a unique function of the advancing contact angle, thetaA, drop size, and material properties (surface tensions and densities). We test our prediction with published and new data. The theory agrees with the experiments. From the relation of the as-placed contact angle to drop size the thermodynamic equilibrium contact angle is also calculated. PMID- 17920073 TI - Cell permeabilization for the assessment of T lymphocyte polyfunctional capacity. AB - The rapid progress in flow cytometry means that an increasing number of parameters can be looked at simultaneously, which is highly relevant for the assessment of lymphocyte characteristics, in particular their multifunctional profile. However, procedures using less complex technology need now optimization in order to take into account the new reagents and applications related to polychromatic flow cytometry. Through optimization of an immunomonitoring protocol to assess the functional profile of antigen specific T-cells using 9-10 colour flow cytometry, we tested the efficacy of three distinct standardized permeabilization buffers for the staining of relevant intracellular molecules. We show significant discrepancies in staining sensitivity for cytokine and cytotoxic factor expression from one permeabilization kit to another, which can lead to different data and interpretation. It is important to be aware of this potential bias and to design specific application/experimental procedures in order to obtain optimal results. PMID- 17920072 TI - Characterization of inflammatory cell infiltration in feline allergic skin disease. AB - Sixteen cats with allergic dermatitis and six control cats with no skin disease were examined. Lymphoid and histiocytic cells in skin sections were examined immunohistochemically and mast cells were identified by toluidine blue staining. The 16 allergic cats showed one or more of several features (alopecia, eosinophilic plaques or granulomas, papulocrusting lesions), and histopathological findings were diverse. In control cats there were no cells that expressed IgM or MAC387, a few that were immunolabelled for IgG, IgA or CD3, and moderate numbers of mast cells. In allergic cats, positively labelled inflammatory cells were generally more numerous in lesional than in non-lesional skin sections, and were particularly associated with the superficial dermis and perifollicular areas. There were low numbers of plasma cells expressing cytoplasmic immunoglobulin; moderate numbers of MHC II-, MAC387- and CD3-positive cells; and moderate to numerous mast cells. MHC class II expression was associated with inflammatory cells morphologically consistent with dermal dendritic cells and macrophages, and epidermal Langerhans cells. Dendritic cells expressing MHC class II were usually associated with an infiltrate of CD3 lymphocytes, suggesting that these cells participate in maintenance of the local immune response by presenting antigen to T lymphocytes. These findings confirm that feline allergic skin disease is characterized by infiltration of activated antigen-presenting cells and T lymphocytes in addition to increased numbers of dermal mast cells. This pattern mimics the dermal inflammation that occurs in the chronic phase of both canine and human atopic dermatitis. PMID- 17920074 TI - Heme-oxygenase-1-induced protection against hypoxia/reoxygenation is dependent on biliverdin reductase and its interaction with PI3K/Akt pathway. AB - Heme-oxygenase-1 (HO-1), a stress-inducible protein, is an important cytoprotective agent against ischemia/reperfusion (I/R) injury. However, the role of downstream mediators involved in HO-1-induced cytoprotection is not clear. In the current study we investigated the role of biliverdin reductase, an enzyme involved in the conversion of HO-1-derived biliverdin into bilirubin and the PI3K/Akt pathway in mediating the cytoprotective effects of HO-1 against hypoxia and reoxygenation (H/R) injury in vitro and in vivo. H9c2 cardiomyocytes were transfected with a plasmid expressing HO-1 or LacZ and exposed to 24 h of hypoxia followed by 12 h of reoxygenation. At the end of reoxygenation, reactive oxygen species generation was determined using CM-H(2)DCFDA dye and apoptosis was assessed by TUNEL, caspase activity and Bad phosphorylation. p85 and Akt phosphorylation were determined using cell-based ELISA and phospho-specific antibodies, respectively. HO-1 overexpression increased phosphorylation of the regulatory subunit of the PI3K (p85alpha) and downstream effector Akt in H9c2 cells, leading to decreased ROS and apoptosis. Furthermore, cardiac expression of HO-1 increased basal phosphorylated Akt levels and decreased infarct size in response to LAD ligation and release induced I/R injury. Conversely, PI3K inhibition reversed the effects of HO-1 on Akt phosphorylation, cell death and infarct size. In addition, knockdown of biliverdin reductase (BVR) expression with siRNA attenuated HO-1-induced Akt phosphorylation and increased H/R-induced apoptosis of H9c2 cells. Co-immunoprecipitation revealed protein-protein interaction between BVR and the phosphorylated p85 subunit of the PI3 kinase. Taken together, these results suggest that the enzyme biliverdin reductase plays an important role in mediating cytoprotective effects of HO-1. This effect is mediated, at least in part, via interaction with and activation of the PI3K/Akt pathway. PMID- 17920076 TI - Cross-talk between iron and nitrogen regulatory networks in anabaena (Nostoc) sp. PCC 7120: identification of overlapping genes in FurA and NtcA regulons. AB - Nitrogen signalling in cyanobacteria involves a complex network in which the availability of iron plays an important role. In the nitrogen-fixing cyanobacterium Anabaena sp. PCC 7120, iron uptake is controlled by FurA, while NtcA is the master regulator of nitrogen metabolism and shows a mutual dependence with HetR in the first steps of heterocyst development. Expression of FurA is modulated by NtcA and it is enhanced in a hetR(-) background. Iron starvation in cells grown in the presence of combined nitrogen causes a moderate increase in the transcription of glnA that is more evident in a ntcA(-) background. Those results evidence a tight link between the reserves of iron and nitrogen metabolism that leads us to search for target genes potentially co-regulated by FurA and NtcA. Using a bioinformatic approach we have found a significant number of NtcA-regulated genes exhibiting iron boxes in their upstream regions. Our computational predictions have been validated using electrophoretic mobility shift assay (EMSA) analysis. These candidates for dual regulation are involved in different functions such as photosynthesis (i.e. psaL, petH, rbcL, isiA), heterocyst differentiation (i.e. xisA, hanA, prpJ, nifH), transcriptional regulation (several alternative sigma factors) or redox balance (i.e. trxA, ftrC, gor). The identification of common elements overlapping the NtcA and FurA regulons allows us to establish a previously unrecognized transcriptional regulatory connection between iron homeostasis, redox control and nitrogen metabolism. PMID- 17920077 TI - Vanadium induces VL30 retrotransposition at an unusually high level: a possible carcinogenesis mechanism. AB - Carcinogenesis by vanadium is thought to occur through induction of DNA-double strand breaks (DSBs) but its mechanism is not fully understood. We investigated the effect of vanadium on induction of viral-like 30 element (VL30) retrotransposition using a NIH3T3 cell-retrotransposition assay based on a recombinant VL30/EGFP element. Incubation of assay cells with vanadyl sulphate (VOSO(4)) induced retrotransposition frequency in a dose and time-dependent manner, measured by fluorescence-activated cell scanning (FACS) and retrotransposition events were confirmed by UV microscopy and PCR analysis. Among vanadium salts with different valence tested, vanadyl (4+) ions were the most potent retrotransposition inducers. VOSO(4), at 50 muM induced retrotranspositions at an unusually high frequency of up to 0.185 events per cell per generation. VOSO(4), acting at the transcription level, strongly induced VL30 and endogenous reverse transcriptase (enRT) transcripts with maxima at 50 muM and 100 muM of 22 and 18-fold, respectively. VOSO(4)-induced retrotransposition frequency was inhibited by 42% with efavirenz, an inhibitor of enRTs, while paraquat, a DNA-DSBs inducer, had no effect. Furthermore, it was completely abolished with deferoxamine, a metal chelator, while reduced by 75% with N-acetyl cysteine, a general antioxidant. Remarkably, H(2)O(2) reproduced inducible retrotransposition linking for the first time oxidative stress to induction of retrotransposition. We propose that VOSO(4)-induced VL30 retrotransposition through H(2)O(2) generation may be an alternative mutagenic, DNA-DSBs independent, mechanism leading to carcinogenesis. PMID- 17920075 TI - Site-directed alkylation of LacY: effect of the proton electrochemical gradient. AB - Previous N-ethylmaleimide-labeling studies show that ligand binding increases the reactivity of single-Cys mutants located predominantly on the periplasmic side of LacY and decreases reactivity of mutants located for the most part of the cytoplasmic side. Thus, sugar binding appears to induce opening of a periplasmic pathway with closing of the cytoplasmic cavity resulting in alternative access of the sugar-binding site to either side of the membrane. Here we describe the use of a fluorescent alkylating reagent that reproduces the previous observations with respect to sugar binding. We then show that generation of an H(+) electrochemical gradient (Delta(mu (H)+), interior negative) increases the reactivity of single-Cys mutants on the periplasmic side of the sugar-binding site and in the putative hydrophilic pathway. The results suggest that Delta(mu (H)+), like sugar, acts to increase the probability of opening on the periplasmic side of LacY. PMID- 17920079 TI - Homotopic dynamics of telomeric loops in mammalian chromatids. AB - Asymmetric transfer of a solitonic kink between telomeric loops shared by template-replica chromatids is conceptualized in terms of homotopic dynamics. Downscaled to B-Z DNA these ideas lead to interesting possibilities regarding duplication and/or selective activation of genes located at or near DNA "hybrid" chiralities. PMID- 17920078 TI - Computerized measurement of negative symptoms in schizophrenia. AB - Accurate measurement of negative symptoms is crucial for understanding and treating schizophrenia. However, current measurement strategies are reliant on subjective symptom rating scales, which often have psychometric and practical limitations. Computerized analysis of patients' speech offers a sophisticated and objective means of evaluating negative symptoms. The present study examined the feasibility and validity of using widely-available acoustic and lexical-analytic software to measure flat affect, alogia and anhedonia (via positive emotion). These measures were examined in their relationships to clinically-rated negative symptoms and social functioning. Natural speech samples were collected and analyzed for 14 patients with clinically-rated flat affect, 46 patients without flat affect and 19 healthy controls. The computer-based inflection and speech rate measures significantly discriminated patients with flat affect from controls, and the computer-based measure of alogia and negative emotion significantly discriminated the flat and nonflat patients. Both the computer and clinical measures of positive emotion/anhedonia corresponded to functioning impairments. The computerized method of assessing negative symptoms offered a number of advantages over the symptom scale-based approach. PMID- 17920080 TI - Roles of chymase in stenosis occurring after polytetrafluoroethylene graft implantations. AB - Chymase is an important enzyme for the generation of angiotensin (Ang) II and in the activation of transforming growth factor (TGF)-beta1. Therefore, chymase may be involved in the hemodialysis access dysfunction, which is caused by intimal hyperplasia that occurs after polytetrafluoroethylene (PTFE) graft implantations. Bilateral U-shaped PTFE grafts were placed between the femoral vein and artery in dogs. Chymase inhibitor (NK3201, 1 mg/kg per day, p.o.) treatments were initiated 3 days before the operation. After the implantation, the stenosis by neointima proliferation was most frequently observed in the venous side of the PTFE grafts. In the hyperplastic neointima, myofibroblasts were the main cellular components. On the other hand, fibroblasts only occupied cellular components in a much smaller proportion in the neointima. However, these cells seem to be rich in the properties of proliferation and migration. After PTFE graft implantations, extensive accumulations of chymase-positive mast cells were found mainly in the tissue surrounding the grafts. The Ang II- and TGF-beta-positive cells were found in an adjacent section that was in close proximity to the chymase-positive cells. In contrast, the AT(1) receptors, as well as TGF-beta type II receptors, were expressed either in the neointima or in the outside adventitia of the PTFE grafts. Chymase inhibitor treatment resulted in a reduction of chymase, Ang II and TGF-beta1 expression, leading to a significant inhibition of neointimal formation. These findings indicating that an increase of chymase via promoting Ang II and TGF-beta1 generation plays a pivotal role in the neointimal formation after the implantation of PTFE grafts and also suggesting that chymase inhibition may be a new strategy that can be used to prevent PTFE graft dysfunctions in clinical settings. PMID- 17920081 TI - A growth and division model for retinoblastoma. AB - A stochastic growth and division model for studying a two hit cancer is developed and applied to retinoblastoma. Retinoblastoma occurs if both genes coding for a tumor suppressor protein on homologous chromosomes become defective. Germinal cases occur when a patient or carrier, born with one defective gene, suffers a second insult to any progeny retinal cell. Somatic cases are far less likely as two hits to the same cell during development are required. Details of the disease, germinal or somatic, unilateral or bilateral, in combination with case data allow for the estimation of the two parameters of the model: mutation rate, estimated at p=7x10(-7) per chromosome per cell division, and carrier frequency, estimated at f=40 per million. The model indicates that carriers of the disease arise from similar mutations to germ cells; in particular, heridary transmission can occur for only a generation or two before dying out. The results show that a stochastic simulation of a multi-hit cancer is feasible and may predict tumor growth dynamics. A simulation run will have to consist of a few million cells in order to observe even a small number of mutations. And several dozens such runs will have to be simulated. PMID- 17920083 TI - Inhibition of voltage-gated channel currents in rat auditory cortex neurons by salicylate. AB - Salicylate is a medicine for anti-inflammation with a side effect of tinnitus. To understand the mechanisms of tinnitus induced by salicylate, we studied the effects of salicylate on voltage-gated ion channels and action potential firing rates in freshly dissociated rat pyramidal neurons in auditory cortex (AC) using the whole-cell patch technique. We found that salicylate reduced the voltage gated sodium current (I(Na)), the delayed rectifier potassium current (I(K(DR))) and the L-type voltage-gated calcium current (I(Ca,L)) in concentration-dependent manner. An amount of 1mM salicylate shifted the steady-state inactivation curve of I(Na) negatively by about 5mV, shifted the steady-state activation and inactivation curve of I(K(DR)) negatively by approximately 14mV and 17mV, respectively, and shifted the steady-state activation curve of I(Ca,L) negatively by about 10mV. 1mM salicylate significantly increased the action potential firing rates, ultimately. From the results, we speculated that through affecting the voltage-gated ion channels in AC, an important position in auditory system, salicylate increased the firing rate of neurons and enhanced neuronal excitability on the one hand, increased the excitatory transmitters release and reduced the inhibitory transmitter release on the other hand, thus finally induced tinnitus. PMID- 17920082 TI - Decreased withdrawal symptoms but normal tolerance to nicotine in mice null for the alpha7 nicotinic acetylcholine receptor subunit. AB - Withdrawal symptoms are a major deterrent when people try to quit smoking. The alpha7 subunit of the neuronal nicotinic acetylcholine receptor (nAChR) is highly expressed in the brain, and has been suspected to play a major role in nicotine addiction. We studied the influence of alpha7-containing nAChRs on nicotine withdrawal and tolerance, in wild type mice and mice null for the alpha7 nAChR subunit (alpha7 -/-). For withdrawal experiments, animals were implanted with osmotic minipumps delivering nicotine for 13 days. A single intraperitoneal injection of the nAChR antagonists mecamylamine (MEC) or methyllycaconitine (MLA) was used to precipitate withdrawal. In wild type mice, both MEC- and MLA precipitated somatic signs of withdrawal such as increased grooming, scratching and shaking. In alpha7 -/- mice, the somatic effects of MEC-precipitated nicotine withdrawal were significantly reduced. Interestingly, the presumed alpha7 specific antagonist MLA also precipitated withdrawal. Tolerance, which was measured as a decrease in nicotine-induced hypolocomotion after subchronic nicotine treatment, was normal in alpha7 -/- mice. Finally, because anxiety and withdrawal symptoms are highly correlated in humans, we studied anxiety-like behaviors in alpha7 -/- mice using a battery of anxiety-related tests. The behavior of alpha7 -/- mice was indistinguishable from that of control mice. Our results point to the alpha7 subunit as one of the players in nicotine withdrawal, but not in nicotine tolerance or basal anxiety-like behavior. PMID- 17920085 TI - Categorical and thematic knowledge representation in the brain: neural correlates of taxonomic and thematic conceptual relations. AB - Most current models of knowledge organization in the brain are based on hierarchical or taxonomic categories (animals, tools). Another important organizational pattern is thematic categorization, i.e. categories held together by external relations, a unifying scene or event (car and garage). We used fMRI to examine neural activation patterns as subjects performed a category construction task where these two category types were contrasted. Subjects were visually presented with a target word followed by the presentation of two match words and had to choose by button press one match that goes best with the target word. In the balanced or cross-categorization condition (Car/Garage Bus) both match words fit the target; in the biased conditions only one match word fit the target either thematically (Car/Garage Brush) or taxonomically (Car/Bus Eraser). We found that in the biased conditions, thematic and taxonomic categories recruited very similar cortical regions: left inferior frontal, middle temporal and occipital regions. In the balanced condition subjects showed no behavioral preference for either thematic or taxonomic matches. However, contrasting signal changes during a subjective taxonomic choice in the presence of a thematic alternative vs. a subjective thematic choice in the presence of a taxonomic alternative required the additional recruitment of right middle frontal gyrus, left precuneus and left thalamus. Our results suggest that thematic relations between objects are processed similarly to taxonomic relations, but require less cerebral processing demand, providing validation for thematic categories as an alternative principle of conceptual organization. PMID- 17920084 TI - Standard versus tailored left temporal lobe resections: differences in cognitive outcome? AB - This study explores differences in cognitive outcome after a standard resection (SR) or tailored (TR) in 100 patients with left temporal lobe epilepsy, controlling for extent in the three lateral gyri. Comparing preoperative to 6 month postoperative performance on a battery of intelligence, language and verbal memory tests revealed the following: a differential effect of the procedure was found for digit span, a short-term memory and attention task, the SR group showing a gain and the TR group a loss postoperatively. This could be explained by a rather large improvement of the SR group with below average resection sizes in the superior temporal gyrus (STG) (<2.8 cm), which small resections are nearly absent in TR resections. Effect of larger extent on the STG in the SR group was related to a decrease in verbal intelligence and a tendency in auditory comprehension which poses a risk in 'large' standard resections. Differences in extent of resection on the other gyri did not cause differences in effects on language functioning or verbal memory. CONCLUSIONS: In standard anterior temporal lobe resections only (without intraoperative language mapping) up to a limit of 4.5 cm, large resections on the STG pose a risk for declining on verbal IQ and auditory comprehension. In general, tailored resections (with language mapping) result in decline on a task measuring short-term memory and attention. PMID- 17920086 TI - Interhemispheric transfer and integration of imagined visual stimuli. AB - We employed two reaction time paradigms to find out whether imagined visual stimuli can be integrated between the two cerebral hemispheres. In a first experiment we found that interhemispheric transfer time, as assessed with the Poffenberger paradigm, was much longer for imagined than visible stimuli and this suggests that the callosal site of transfer is different in the two conditions. In a second experiment we found that interhemispheric summation, as assessed with the redundant signal effect paradigm, was present for both visible and imagined stimuli and could be accounted for by a neural coactivation mechanism rather than by a probabilistic explanation. Taken together, these results support the view that that there is an equivalence between perceptual and imagery processes that goes beyond early processing stages and includes the interhemispheric exchange of information. PMID- 17920089 TI - Modification of hemicellulose content by antisense down-regulation of UDP glucuronate decarboxylase in tobacco and its consequences for cellulose extractability. AB - Extractability and recovery of cellulose from cell walls influences many industrial processes and also the utilisation of biomass for energy purposes. The utility of genetic manipulation of lignin has proven potential for optimising such processes and is also advantageous for the environment. Hemicelluloses, particularly secondary wall xylans, also influence the extractability of cellulose. UDP-glucuronate decarboxylase produces UDP-xylose, the precursor for xylans and the effect of its down-regulation on cell wall structure and cellulose extractability in transgenic tobacco has been investigated. Since there are a number of potential UDP-glucuronate decarboxylase genes, a 490bp sequence of high similarity between members of the family, was chosen for general alteration of the expression of the gene family. Sense and antisense transgenic lines were analysed for enzyme activity using a modified and optimised electrophoretic assay, for enzyme levels by western blotting and for secondary cell wall composition. Some of the down-regulated antisense plants showed high glucose to xylose ratios in xylem walls due to less xylose-containing polymers, while arabinose and uronic acid contents, which could also have been affected by any change in UDP-xylose provision, were unchanged. The overall morphology and stem lignin content of the modified lines remained little changed compared with wild type. However, there were some changes in vascular organisation and reduction of xylans in the secondary walls was confirmed by immunocytochemistry. Pulping analysis showed a decreased pulp yield and a higher Kappa number in some lines compared with controls, indicating that they were less delignified, although the level of residual alkali was reduced. Such traits probably indicate that lignin was less available for removal in a reduced background of xylans. However, the viscosity was higher in most antisense lines, meaning that the cellulose was less broken-down during the pulping process. This is one of the first studies of a directed manipulation of hemicellulose content on cellulose extractability and shows both positive and negative outcomes. PMID- 17920088 TI - ESP and ESM1 mediate indol-3-acetonitrile production from indol-3-ylmethyl glucosinolate in Arabidopsis. AB - Glucosinolates are plant secondary metabolites that act as direct defenses against insect herbivores and various pathogens. Recent analysis has shown that methionine-derived glucosinolates are hydrolyzed/activated into either nitriles or isothiocyanates depending upon the plants genotype at multiple loci. While it has been hypothesized that tryptophan-derived glucosinolates can be a source of indole-acetonitriles, it has not been explicitly shown if the same proteins control nitrile production from tryptophan-derived glucosinolates as from methionine-derived glucosinolates. In this report, we formally test if the proteins involved in controlling aliphatic glucosinolate hydrolysis during tissue disruption can control production of nitriles during indolic glucosinolate hydrolysis. We show that myrosinase is not sufficient for indol-3-acetonitrile production from indol-3-ylmethyl glucosinolate and requires the presence of functional epithospecifier protein in planta and in vitro to produce significant levels of indol-3-acetonitrile. This reaction is also controlled by the Epithiospecifier modifier 1 gene. Thus, like formation of nitriles from aliphatic glucosinolates, indol-3-acetonitrile production following tissue disruption is controlled by multiple loci raising the potential for complex regulation and fine tuning of indol-3-acetonitrile production from indol-3-ylmethyl glucosinolate. PMID- 17920087 TI - Overlapping numerical cognition impairments in children with chromosome 22q11.2 deletion or Turner syndromes. AB - Children with one of two genetic disorders (chromosome 22q11.2 deletion syndrome and Turner syndrome) as well typically developing controls, participated in three cognitive processing experiments. Two experiments were designed to test cognitive processes involved in basic aspects numerical cognition. The third was a test of simple manual motor reaction time. Despite significant differences in global intellectual abilities, as measured by IQ tests, performance on the two numerical cognition tasks differed little between the two groups of children with genetic disorders. However, both performed significantly more poorly than did controls. The pattern of results are consistent with the hypothesis that impairments were not due to global intellectual ability but arose in specific cognitive functions required by different conditions within the tasks. The fact that no group differences were found in the reaction time task, despite significant differences in the standardized processing speed measure, further supports the interpretation that specific cognitive processing impairments and not global intellectual or processing speed impairments explain the pattern of results. The similarity in performance on these tasks of children with unrelated genetic disorders counters the view that numerical cognition is under any direct genetic control. Instead, our findings are consistent with the view that disturbances in foundational spatiotemporal cognitive functions contribute to the development of atypical representations and processes in the domains of basic magnitude comparison and simple numerical enumeration. PMID- 17920090 TI - Antioxidant phenylpropanoid glycosides from the leaves of Wasabia japonica. AB - From the MeOH extract of the leaves of W. japonica, seven phenylpropanoid gentiobiosides (1-7) were isolated along with eight known phenylpropanoids (8 15). Structures of 1-7 were determined based on spectroscopic data and chemical evidence. The activity of compounds 1-15 to scavenge superoxide anion radicals was investigated using an electron spin resonance (ESR) method. PMID- 17920091 TI - Influence of electron transport proteins on the reactions catalyzed by Fusarium fujikuroi gibberellin monooxygenases. AB - The multifunctional cytochrome P450 monooxygenases P450-1 and P450-2 from Fusarium fujikuroi catalyze the formation of GA14 and GA4, respectively, in the gibberellin (GA)-biosynthetic pathway. However, the activity of these enzymes is qualitatively and quantitatively different in mutants lacking the NADPH:cytochrome P450 oxidoreductase (CPR) compared to CPR-containing strains. 3beta-Hydroxylation, a major P450-1 activity in wild-type strains, was strongly decreased in the mutants relative to oxidation at C-6 and C-7, while synthesis of C19-GAs as a result of oxidative cleavage of C-20 by P450-2 was almost absent whereas the C-20 alcohol, aldehyde and carboxylic acid derivatives accumulated. Interaction of the monooxygenases with alternative electron transport proteins could account for these different product distributions. In the absence of CPR, P450-1 activities were NADH-dependent, and stimulated by cytochrome b5 or by added FAD. These properties as well as the decreased efficiency of P450-1 and P450-2 in the mutants are consistent with the participation of cytochrome b5:NADH cytochrome b5 reductase as redox partner of the gibberellin monooxygenases in the absence of CPR. We provide evidence, from either incubations of GA12 (C-20 methyl) with cultures of the mutant suspended in [18O]H2O or maintained under an atmosphere of [18O]O2:N2 (20:80), that GA15 (C-20 alcohol) and GA24 (C-20 aldehyde) are formed directly from dioxygen and not from hydrolysis of covalently enzyme-bound intermediates. Thus these partially oxidized GAs correspond to intermediates of the sequential oxidation of C-20 catalyzed by P450-2. PMID- 17920092 TI - Argemone mexicana decoction for the treatment of uncomplicated falciparum malaria. AB - A prospective, dose-escalating, quasi-experimental clinical trial was conducted with a traditional healer using a decoction of Argemone mexicana for the treatment of malaria in Mali. The remedy was prescribed in three regimens: once daily for 3 days (Group A; n=23); twice daily for 7 days (Group B; n=40); and four times daily for the first 4 days followed by twice daily for 3 days (Group C; n=17). Thus, 80 patients were included, of whom 80% were aged<5 years and 25% were aged<1 year. All presented to the traditional healer with symptoms of malaria and had a Plasmodium falciparum parasitaemia>2000/microl but no signs of severe malaria. The proportions of adequate clinical response (ACR) at Day 14 were 35%, 73% and 65% in Groups A, B and C, respectively (P=0.011). At Day 14, overall proportions of ACR were lower in children aged<1 year (45%) and higher in patients aged>5 years (81%) (P=0.027). Very few patients had complete parasite clearance, but at Day 14, 67% of patients with ACR had a parasitaemia<2000/microl. No patient needed referral for severe disease. Only minor side effects were observed. Further research should determine whether this local resource could represent a first-aid home treatment in remote areas. PMID- 17920093 TI - Dynamics of Fst for the island model. AB - F(st) is a measure of genetic differentiation in a subdivided population. Sewall Wright observed that F(st)=1/1+2Nm in a haploid diallelic infinite island model, where N is the effective population size of each deme and m is the migration rate. In demonstrating this result, Wright relied on the infinite size of the population. Natural populations are not infinite and therefore they change over time due to genetic drift. In a finite population, F(st) becomes a random variable that evolves over time. In this work we ask, given an initial population state, what are the dynamics of the mean and variance of F(st) under the finite island model? In application both of these quantities are critical in the evaluation of F(st) data. We show that after a time of order N generations the mean of F(st) is slightly biased below 1/1+2Nm. Further we show that the variance of F(st) is of order 1/d where d is the number of demes in the population. We introduce several new mathematical techniques to analyze coalescent genealogies in a dynamic setting. PMID- 17920094 TI - Epigenetic changes in the rat livers induced by pyrazinamide treatment. AB - Drug-induced liver injury, including drug-induced hepatotoxicity during the treatment of tuberculosis infection, is a major health problem with increasingly significant challenges to modern hepatology. Therefore, the assessment and monitoring of the hepatotoxicity of antituberculosis drugs for prevention of liver injury are great concerns during disease treatment. The recently emerged data showing the ability of toxicants, including pharmaceutical agents, to alter cellular epigenetic status, open a unique opportunity for early detection of drug hepatotoxicity. Here we report that treatment of male Wistar rats with antituberculosis drug pyrazinamide at doses of 250, 500 or 1000 mg/kg/day body weight for 45 days leads to an early and sustained decrease in cytosine DNA methylation, progressive hypomethylation of long interspersed nucleotide elements (LINE-1), and aberrant promoter hypermethylation of placental form glutathione-S transferase (GSTP) and p16(INK4A) genes in livers of pyrazinamide-treated rats, while serum levels of bilirubin and activity of aminotransferases changed modestly. The early occurrence of these epigenetic alterations and their association with progression of liver injury specific pathological changes indicate that alterations in DNA methylation may be useful predictive markers for the assessment of drug hepatotoxicity. PMID- 17920095 TI - Selective induction of cell-mediated immunity and protection of rhesus macaques from chronic SHIV(KU2) infection by prophylactic vaccination with a conserved HIV 1 envelope peptide-cocktail. AB - Infection of Indian-origin rhesus macaques by the simian human immunodeficiency virus (SHIV) is considered to be a suitable preclinical model for directly testing efficacy of vaccine candidates based on the HIV-1 envelope. We used this model for prophylactic vaccination with a peptide-cocktail comprised of highly conserved HIV-1 envelope sequences immunogenic/antigenic in macaques and humans. Separate groups of macaques were immunized with the peptide-cocktail by intravenous and subcutaneous routes using autologous dendritic cells (DC) and Freund's adjuvant, respectively. The vaccine elicited antigen specific IFN-gamma producing cells and T-cell proliferation, but not HIV-neutralizing antibodies. The vaccinated animals also exhibited efficient cross-clade cytolytic activity against target cells expressing envelope proteins corresponding to HIV-1 strains representative of multiple clades that increased after intravenous challenge with pathogenic SHIV(KU2). Virus-neutralizing antibodies were either undetectable or present only transiently at low levels in the control as well as vaccinated monkeys after infection. Significant control of plasma viremia leading to undetectable levels was achieved in majority of vaccinated monkeys compared to mock-vaccinated controls. Monkeys vaccinated with the peptide-cocktail using autologous DC, compared to Freund's adjuvant, and the mock-vaccinated animals, showed significantly higher IFN-gamma production, higher levels of vaccine specific IFN-gamma producing CD4(+) cells and significant control of plasma viremia. These results support DC-based vaccine delivery and the utility of the conserved HIV-1 envelope peptide-cocktail, capable of priming strong cell mediated immunity, for potential inclusion in HIV vaccination strategies. PMID- 17920096 TI - Comparative requirements for the restriction of retrovirus infection by TRIM5alpha and TRIMCyp. AB - The restriction factors, TRIM5alpha in most primates and TRIMCyp in owl monkeys, block infection of various retroviruses soon after virus entry into the host cell. Rhesus monkey TRIM5alpha (TRIM5alpha rh) inhibits human immunodeficiency virus (HIV-1) and feline immunodeficiency virus (FIV) more potently than human TRIM5alpha (TRIM5alpha hu). TRIMCyp restricts infection of HIV-1, simian immunodeficiency virus of African green monkeys (SIV agm) and FIV. Early after infection, TRIMCyp, like TRIM5alpha rh and TRIM5alpha hu, decreased the amount of particulate viral capsid in the cytosol of infected cells. The requirements for the TRIMCyp and TRIM5alpha domains in restricting different retroviruses were investigated. Potent restriction of FIV by TRIMCyp occurred in the complete absence of RING and B-box 2 domains; by contrast, efficient FIV restriction by TRIM5alpha rh required these domains. Variable region 1 of the TRIM5alpha rh B30.2 domain contributed to the potency of HIV-1, FIV and equine infectious anemia virus restriction. Thus, although differences exist in the requirements of TRIMCyp and TRIM5alpha for RING/B-box 2 domains, both restriction factors exhibit mechanistic similarities. PMID- 17920098 TI - Selectivity for coherence in polar orientation in human form vision. AB - Evidence from summation near threshold psychophysical experiments using compound Glass patterns is presented which supports the existence of mechanisms in the human visual system tuned for coherence in radial and concentric, and +45 degrees and -45 degrees spiral orientations. It is suggested that sensitivity to +45 degrees and -45 degrees logarithmic spirals serves to disambiguate the sense of spiral form, which would not be uniquely specified by measures of the components of orientation along the radial and concentric directions alone. A spiral space is introduced within which radial and concentric patterns are diametrically opposed on one axis and spirals of +45 degrees and -45 degrees on an orthogonal axis and it is proposed that these represent cardinal axes for detecting global structure. Comparison of the sensitivity tuning functions of the four mechanisms tuned to these axes with sensitivity to simple spiral Glass patterns shows that weighted combinations of output from adjacent pairs of this set of mechanisms are sufficient to account for absolute sensitivity to logarithmic spiral Glass patterns of all intermediate spiral angles. Control experiments demonstrate that the combinations are labeled for spiral sense (simple spirals of -22.5 degrees spiral angle can be discriminated from +22.5 degrees spirals at threshold for detection) and that adaptation transfers across quadrants of spiral space (adaptation to spirals of -22.5 degrees results in a decrease in sensitivity to orthogonal +22.5 degrees and -67.5 degrees spirals). Together these observations suggest that sensitivity to spirals in each of the quadrants of spiral space is due to higher order mechanisms reliant on output from 0 degrees , 90 degrees , +45 degrees and -45 degrees cardinal mechanisms. PMID- 17920097 TI - Analysis of gene expression in fetal and adult cells infected with rubella virus. AB - Congenital infection with rubella virus (RUB) leads to persistent infection and congenital defects and we showed previously that primary human fetal fibroblasts did not undergo apoptosis when infected with RUB, which could promote fetal virus persistence [Adamo, P., Asis, L., Silveyra, P., Cuffini, C., Pedranti, M., Zapata, M., 2004. Rubella virus does not induce apoptosis in primary human embryo fibroblasts cultures: a possible way of viral persistence in congenital infection. Viral Immunol. 17, 87-100]. To extend this observation, gene chip analysis was performed on a line of primary human fetal fibroblasts (10 weeks gestation) and a line of human adult lung fibroblasts (which underwent apoptosis in response to RUB infection) to compare gene expression in infected and uninfected cells. A total of 632 and 516 genes were upregulated or downregulated in the infected fetal and adult cells respectively in comparison to uninfected cells, however only 52 genes were regulated in both cell types. Although the regulated genes were different, across functional gene categories the patterns of gene regulation were similar. In general, regulation of pro- and anti-apoptotic genes following infection appeared to favor apoptosis in the adult cells and lack of apoptosis in the fetal cells, however there was a greater relative expression of anti-apoptotic genes and reduced expression of pro-apoptotic genes in uninfected fetal cells versus uninfected adult cells and thus the lack of apoptosis in fetal cells following RUB infection was also due to the prevailing background of gene expression that is antagonistic to apoptosis. In support of this hypothesis, it was found that of a battery of five chemicals known to induce apoptosis, two induced apoptosis in the adult cells, but not in fetal cells, and two induced apoptosis more rapidly in the adult cells than in fetal cells (the fifth did not induce apoptosis in either). A robust interferon-stimulated gene response was induced following infection of both fetal and adult cells and many of the genes upregulated in both cell types were those involved in establishment of an antiviral state; this is the first demonstration of an interferon response at this early stage of human embryonic development. In both fetal and adult cells, interferon controlled but did not eliminate virus spread and apoptosis was not induced in infected fetal cells in the absence of interferon. In addition to the interferon response, chemokines were induced in both infected fetal and adult cells. Thus, it is possible that fetal damage following congenital RUB infection, which involves cell proliferation and differentiation, could be due to induction of the innate immune response as well as frank virus infection. PMID- 17920099 TI - Broadly tuned, view-specific coding of face shape: opposing figural aftereffects can be induced in different views. AB - Face aftereffects are sensitive to changes in viewpoint, suggesting view-specific face coding, yet are not entirely eliminated by changes in viewpoint, suggesting view-invariance. To determine whether broad view-tuning can account for these findings we measured the reduction of a figural face aftereffect induced in one view by concurrent adaptation to an opposite distortion in a second viewpoint, varying the angle between these views. To the degree that the same neural population codes both viewpoints, the opposing aftereffects should cancel. Cancellation increased monotonically as the angle between two adapting views decreased, consistent with broadly tuned, view-specific coding of face shape. PMID- 17920100 TI - Dynamics of nitric oxide and nitrous oxide emission during full-scale reject water treatment. AB - Emission of NO and N2O from a full-scale two-reactor nitritation-anammox process was determined during a measurement campaign at the Dokhaven-Sluisjesdijk municipal WWTP (Rotterdam, NL). The NO and N2O levels in the off-gas responded to the aeration cycles and the aeration rate of the nitritation reactor, and to the nitrite and dissolved oxygen concentration. Due to the strong fluctuations in the NO and N2O levels in both the nitritation and the anammox reactor, only time dependent measurements could yield a reliable estimate of the overall NO and N2O emissions. The NO emission from the nitritation reactor was 0.2% of the nitrogen load and the N2O emission was 1.7%. The NO emission from the anammox reactor was determined to be 0.003% of the nitrogen load and the N2O emission was 0.6%. Emission of NO2 could not be detected from the nitritation-anammox system. Denitrification by ammonia-oxidizing bacteria was considered to be the most probable cause of NO and N2O emission from the nitritation reactor. Since anammox bacteria have not been shown to produce N2O under physiological conditions, it is also suspected that ammonia-oxidizing bacteria contribute most to N2O production in the anammox reactor. The source of NO production in the anammox reactor can be either anammox bacteria or denitrification by heterotrophs or ammonia-oxidizing bacteria. Based on the results and previous work, it seems that a low dissolved oxygen or a high nitrite concentration are the most likely cause of elevated NO and N2O emission by ammonia-oxidizing bacteria. The emission was compared with measurements at other reject water technologies and with the main line of the Dokhaven-Sluisjesdijk WWTP. The N2O emission levels in the reject water treatment seem to be in the same range as for the main stream of activated sludge processes. Preliminary measurements of the N2O emission from a one-reactor nitritation-anammox system indicate that the emission is lower than in two reactor systems. PMID- 17920101 TI - EDDS and EDTA-enhanced zinc accumulation by Solanum nigrum inoculated with arbuscular mycorrhizal fungi grown in contaminated soil. AB - The effect of two different chelating agents [EDTA and EDDS (S,S ethylenediaminedissucinic acid)] on Zn tissue accumulation in Solanum nigrum L. grown in a naturally contaminated soil was assessed. Under those conditions, the response of the plant to the inoculation with two different isolates of arbuscular mycorrhizal fungi (AMF)--Glomus claroideum and Glomus intraradices- was also studied. Plants grown in the local contaminated soil (Zn levels of 433mg kg(-1)) accumulated up to 1191mg kg(-1) of Zn in the roots, 3747mg kg(-1) in the stems and 3409mg kg(-1) in the leaves. S. nigrum plants grown in the same soil spiked with extra Zn (Zn levels of 964mg kg(-1)) accumulated up to 4735, 8267 and 7948mg Zn kg(-1) in the leaves, stems and roots, respectively. The addition of EDTA promoted an increase in the concentration of Zn accumulated by S. nigrum of up to 231% in the leaves, 93% in the stems and 81% in the roots, while EDDS application enhanced the accumulation in leaves, stems and roots up to 140, 124 and 104%, respectively. In the stems, the presence of Zn was predominantly detected in the cortex collenchyma cells, the starch sheath and the internal phloem and xylem parenchyma, and the addition of chelating agents did not seem to have an effect on the localisation of accumulation sites. The devise of a chelate enhanced phytoextraction strategy, using chelating agents and AMF, is discussed. PMID- 17920102 TI - Performance of Gd-doped Ti-based Sb-SnO2 anodes for electrochemical destruction of phenol. AB - The performance of electrodes for the electro-catalytic decomposition of a model pollutant (phenol) was enhanced using Gd-doped Ti/SnO(2)-Sb electrodes prepared by a thermal deposition method. Phenol degradation followed first-order rate kinetics, with the maximum rate achieved using a 2% Gd doping level (molar ratio based on Gd:Sn) for tests conducted over a doping range of 1-10%. The first-order rate constant with 2% Gd was 0.044 min(-1), versus 0.026 min(-1) obtained with the control (plain Ti/SnO(2)-Sb). TOC removal and UV scans revealed that different intermediates were produced for different Gd contents, and that destruction efficiencies of these intermediates also varied with Gd doping levels of 1-5%. Electrodes were characterized by scanning electron microscopy, X-ray diffraction, electron dispersive spectrometry, and X-ray photon-electron spectroscopy. It is suggested that the state of specific active sites on the electrode surface and the oxygen transfer activity at the electrode/electrolyte interface affect the performance of anodes with different compositions. PMID- 17920103 TI - Tracking changing environments using stable carbon isotopes in fossil tooth enamel: an example from the South African hominin sites. AB - The environmental contexts of the karstic hominin sites in South Africa have been established largely by means of faunal associations; taken together these data suggest a trend from relatively closed and more mesic to open, drier environments from about 3 to 1.5 Ma. Vrba argued for a major shift within this trend ca. 2.4 2.6 Ma, an influential proposal that posited links between bovid (and hominin) radiation in Africa and the intensification of Northern Hemisphere Glaciation. Yet faunal approaches often rely on habitat and feeding preferences of modern taxa that may differ from those of their extinct predecessors. Here we explore ways of extending (13)C/(12)C data from fossil mammals beyond denoting "presence" or "absence" of C(4) grasses using the evolution of open environments in South Africa as a case study. To do so we calculated the relative proportions of C(3)-, mixed-, and C(4)-feeding herbivores for all the hominin sites for which we have sufficient data based on (13)C/(12)C analyses of fossil tooth enamel. The results confirm a general trend towards more open environments since 3 Ma, but they also emphasize a marked change to open grassy habitats in the latest Pliocene/early Pleistocene. Mean (13)C/(12)C for large felids also mirrored this trend. PMID- 17920104 TI - Stratigraphy, U-Th chronology, and paleoenvironments at Gladysvale Cave: insights into the climatic control of South African hominin-bearing cave deposits. AB - Gladysvale Cave is one of the few Plio-Pleistocene hominin-bearing cave sites in South Africa that contains a well-stratified cave fill with clastic sediments interspersed with flowstones. The clastic sediments can be divided into units based on the presence of intercalated flowstones, forming flowstone bounded units (FBU). Ten MC-ICP-MS uranium-series dates on several flowstone horizons in the Gladysvale Internal Deposit fan indicate deposition from the late mid-Pleistocene ( approximately 570 ka) to Holocene ( approximately 7 ka) during limited periods of higher effective moisture. Clastic sedimentation occurred during the interceding, presumably more arid, periods. This sequence is not consistent with earlier models for South African caves that simply assumed interglacial sedimentation and glacial erosion. (13)C/(12)C data suggest that flowstone tended to form during periods with higher proportions of C(3) plants in the local vegetation, while clastic sediments reflect higher proportions of C(4) grasses, although this is not always the case. We argue that flowstones are precipitated during periods of higher effective precipitation and restricted cave entrances, while clastic sediments accumulated during periods with more open vegetation. The sedimentary fill of the fossiliferous deposits are, therefore, highly episodic in nature, with large periods of time unlikely to be represented. This has serious implications for the other hominin-bearing caves close by, as these deposits are likely to be similarly episodic. This is especially pertinent when addressing extinction events and reconstructions of paleoenvironments, as large periods of time may be unrecorded. The Gladysvale Cave fill sediments may serve as a climatically forced chronostratigraphic model for these less well-stratified and well-dated Plio-Pleistocene sites. PMID- 17920105 TI - The biokinetics of inorganic cobalt in the human body. AB - This paper reviews information on the biological behavior of inorganic cobalt in humans and laboratory animals and proposes a model of the systemic biokinetics of inorganic cobalt in adult humans. The model was developed as part of an effort to update the models of the International Commission on Radiological Protection (ICRP) for addressing intakes of radionuclides by workers but is also applicable to environmental or medical exposures to inorganic forms of radiocobalt. The model can be used in conjunction with any respiratory, gastrointestinal, or wound model that provides predictions of the time-dependent feed of cobalt to blood. In contrast to the ICRP's current systemic model for cobalt, which is a simple open catenary system, the proposed model is constructed within a physiologically realistic framework that depicts recycling of cobalt between blood and tissues and transfer from blood to excretion pathways. Compared with the ICRP's current model, the proposed model yields similar predictions of whole-body retention but substantially different predictions of the systemic distribution of cobalt as a function of time after uptake to blood. PMID- 17920106 TI - Toxicity of urban highway runoff with respect to storm duration. AB - The toxicity of stormwater runoff during various time-based stages was measured in both grab and composite samples collected from three highly urbanized highway sites in Los Angeles, California between 2002 and 2005. Stormwater runoff samples were tested for toxicity using three freshwater species (the water flea Ceriodaphnia dubia, the fathead minnow Pimephales promelas, and the green algae Pseudokirchneriella subcapitatum) and two marine species (the purple sea urchin Strongylocentrotus purpuratus, and the luminescent bacteria Photobacterium phosphoreum using Microtox. Toxicity results varied substantially throughout the storm events for both freshwater and marine species toxicity tests. In general, however, the first few samples were found to be more toxic compared with those collected during later stages of each storm event. In most cases, more than 40% of the toxicity was associated with the first 20% of discharged runoff volume. Furthermore, on average, 90% of the toxicity was observed during the first 30% of storm duration. Toxicity identification evaluation results found copper and zinc to be the primary cause of toxicity in about 90% of the samples evaluated with these procedures. Surfactants were also found to be the cause of toxicity in less than 10% of the samples. PMID- 17920107 TI - Factors associated with the timing of diagnosis of venous thromboembolism: results from the MASTER registry. AB - INTRODUCTION: Signs and symptoms of venous thromboembolism (VTE) are non-specific and thus can make diagnosis difficult, even for an experienced clinician. We aimed to evaluate the timing of diagnosis of deep vein thrombosis (DVT) and pulmonary embolism (PE) in Italian hospitals and to identify individual and clinical predictors of timely or delayed diagnosis. MATERIAL AND METHODS: MASTER is a multicenter prospective registry of patients with acute DVT and PE. Information on clinical presentation and diagnostic methods, temporary and permanent risk factors, were captured by an electronic data network at the time of the index event. RESULTS: Data on 2047 patients (1024 males), 1505 with DVT and 542 with PE, were analysed. Delayed diagnosis (i.e. more than 10 days from onset of symptoms) was observed in 340 (22.6%) patients with DVT and in 88 (16.2%) with PE, respectively. In DVT patients, factors associated with earlier diagnosis were the presence of multiple signs or symptoms (p=0.014), the presence of pain (p=0.049), and previous venous thrombosis (p=0.016). Neither the presence of other known risk factors nor ongoing prophylaxis influenced the timing of diagnosis. In PE patients, only multiple signs or symptoms at presentation (p=0.014) and the presence of transient risk factors (p=0.001) were significantly associated with earlier diagnosis. CONCLUSIONS: Substantial delays occur when diagnosing both DVT and PE. The severity of presentation, but not patient risk profile are associated with earlier diagnosis, even in patients with signs or symptoms of PE. PMID- 17920108 TI - The prognostic effects of performance status and quality of life scores on progression-free survival and overall survival in advanced ovarian cancer. AB - OBJECTIVE: Performance status (PS) is an important prognostic factor in advanced ovarian cancer. The purpose of this study was to evaluate the prognostic significance of PS and quality of life (QoL) assessment on progression-free survival (PFS) and overall survival (OS) in patients with advanced ovarian cancer. METHODS: We studied Canadian patients participating in an intergroup study in ovarian cancer (NCIC-OV10), which randomized patients to receive either standard chemotherapy using cisplatin/cyclophosphamide or cisplatin/paclitaxel chemotherapy. QoL was assessed using the EORTC quality of life questionnaire (QLQ C30+3). The effects of multiple variables including the relevant clinical variables, PS and QoL scores were analyzed by Cox stepwise regression at baseline and again 3 months after completion of chemotherapy. RESULTS: At baseline and at 3 months after chemotherapy, there were 151 and 93 patients respectively who completed the QLQ-C30+3 questionnaires. Baseline PS, global QoL score and treatment were independent predictors for both PFS and OS. Baseline cognitive functioning score was also an additional independent predictor for OS. At 3 months after completion of chemotherapy global QoL score, PS and grade were significant independent predictors of OS; however, only physical functioning score, emotional functioning score and tumor grade predicted for PFS. CONCLUSIONS: Performance status and global quality of life scores at baseline are prognostic factors in advanced ovarian cancer for both PFS and OS. Higher baseline cognitive functioning scores were also associated with improved survival. Global QoL scores at 3 following completion of chemotherapy proved to be of prognostic significance for OS but not PFS. PMID- 17920109 TI - Extracapsular extension of pelvic lymph node metastases is of prognostic value in carcinoma of the cervix uteri. AB - OBJECTIVES: Pelvic lymph node involvement is a well-recognized prognostic factor in cervical carcinoma (CX). Limited knowledge exists about extranodal extension of the tumor outside the lymph node capsule, i.e. extracapsular spread (ECS). METHODS: Two hundred fifty-six cases of surgically treated CX (FIGO stage IB1 to IIB) with pelvic lymph node involvement were evaluated regarding the occurrence of extranodal spread of the metastatic deposits outside the lymph node capsule (ECS), determined on standardized handled lymphadenectomy specimens, regarding their impact of recurrent disease and overall survival during a median follow-up time of 62 months (95% CI 51-73 months). RESULTS: ECS was seen in 30.9% (79/256) of the cases. The occurrence of ECS showed a significant correlation to advanced stage disease (p=0.02), the number of involved nodes (p<0.001) and the size of metastatic deposits (p<0.01). The 5-year recurrence-free survival rate in patients with ECS was significant lower compared to patients without ECS (59.7% [95% CI: 46.3%-73.2%] versus 67.2% [95% CI: 58.9%-75.5%]; (p=0.04). The 5-year overall survival rate was significant lower in patients with ECS (33.5% [95% CI: 20.6%-46.3%] vs. 60.5% [95% CI: 52.3%-68.6%]; p<0.001). In multivariate analysis, tumor stage, number of involved pelvic nodes, tumor differentiation and ECS were independent prognostic factors. CONCLUSIONS: The results indicate that extracapsular spread (ECS) of pelvic lymph node metastases is of prognostic impact in cervical carcinomas. A revised FIGO/TNM classification system for pelvic lymph node disease is recommended: ECS 0 = lymph node involvement without extranodal spread of the metastatic deposits and ECS 1 = lymph node involvement with extranodal spread of the metastatic deposits. PMID- 17920110 TI - Combining multiple serum tumor markers improves detection of stage I epithelial ovarian cancer. AB - OBJECTIVE: Currently available tumor markers for ovarian cancer are still inadequate in both sensitivity and specificity to be used for population-based screening. Artificial neural network (ANN) as a modeling tool has demonstrated its ability to assimilate information from multiple sources and to detect subtle and complex patterns. In this paper, an ANN model was evaluated for its performance in detecting early stage epithelial ovarian cancer using multiple serum markers. METHODS: Serum specimens collected at four institutions in the US, The Netherlands and the United Kingdom were analyzed for CA 125II, CA 72-4, CA 15 3 and macrophage colony stimulating factor (M-CSF). The four tumor marker values were then used as inputs to an ANN derived using a training set from 100 apparently healthy women, 45 women with benign conditions arising from the ovary and 55 invasive epithelial ovarian cancer patients (including 27 stage I/II cases). A separate validation set from 27 apparently healthy women, 56 women with benign conditions and 35 women with various types of malignant pelvic masses was used to monitor the ANN's performance during training. An independent test data set from 98 apparently healthy women and 52 early stage epithelial ovarian cancer patients (38 stage I and 4 stage II invasive cases and 10 stage I borderline ovarian tumor cases) was used to evaluate the ANN. RESULTS: ROC analysis confirmed the overall superiority of the ANN-derived composite index over CA 125II alone (p=0.0333). At a fixed specificity of 98%, the sensitivities for ANN and CA 125II alone were 71% (37/52) and 46% (24/52) (p=0.047), respectively, for detecting early stage epithelial ovarian cancer, and 71% (30/42) and 43% (18/42) (p=0.040), respectively, for detecting invasive early stage epithelial ovarian cancer. CONCLUSIONS: The combined use of multiple tumor markers through an ANN improves the overall accuracy to discern healthy women from patients with early stage ovarian cancer. Analysis of multiple markers with an ANN may be a better choice than the use of CA 125II alone in a two-step approach for population screening in which a secondary test such as ultrasound is used to keep the overall specificity at an acceptable level. PMID- 17920112 TI - Lifestyle and reduced mortality among active California Mormons, 1980-2004. AB - OBJECTIVE: The objective is to measure the relationship of several healthy characteristics of the Mormon lifestyle to mortality. METHOD: We examined 9815 religiously active California Mormon adults followed for mortality during 1980 2004 and 15,832 representative U.S. white adults enrolled in the 1987 National Health Interview Survey (NHIS) and followed for mortality during 1988-1997. The standardized mortality ratio (SMR) and 95% confidence interval (CI) was calculated relative to U.S. whites defined to have a SMR of 1.00. RESULTS: Active California Mormons practice a healthy lifestyle advocated by their religion, which emphasizes a strong family life, education and abstention from tobacco and alcohol. Unusually low SMRs occurred among married never smokers who attended church weekly and had at least 12 years of education. For those aged 25-99 years at entry, the SMR for all causes of death was 0.45 (0.42-0.48) for males and 0.55 (0.51-0.59) for females. For those aged 25-64 years at entry, the SMR for all causes of death was 0.36 (0.32-0.41) for males and 0.46 (0.40-0.53) for females. Life expectancy from age 25 was 84 years for males and 86 years for females. These SMRs were largely replicated among similarly defined persons of all religions within the NHIS cohort. CONCLUSIONS: Several healthy characteristics of the Mormon lifestyle are associated with substantially reduced death rates and increased life expectancy. PMID- 17920111 TI - In utero methanesulfonyl fluoride differentially affects learning and maze performance in the absence of long-lasting cholinergic changes in the adult rat. AB - There is increasing evidence that acetylcholinesterase (AChE) may have various specific developmental roles in brain development. Nevertheless, specific effects of AChE inhibition during early brain development have not been adequately described. Therefore, methanesulfonyl fluoride (MSF), an irreversible AChE inhibitor that shows high selectivity for the CNS was used to produce AChE inhibition in utero to study subsequent adult behaviors, sleep, and cholinergic markers. Rats exposed to MSF in utero showed a deficit in spatial learning tasks using appetitive motivation but, surprisingly, they performed equally well or better than controls when aversive motivation was used. One hypothesis was that MSF treatment in utero affected the response to stress. Tests of anxiety however showed no differences in basal levels of anxiety. Studies of sleep behavior, however, indicated a higher level of REM sleep which is only seen during the light phase of male rats exposed to MSF in utero as compared to controls. No differences in cholinergic markers in the brains of adults were found except that females exposed to MSF in utero had a higher level of ChAT activity in the synaptosomal fraction of the hippocampus. Even so, whether cholinergic alterations accompany the in utero MSF exposure remains to be determined. The failure to find widespread changes in cholinergic markers in the adult brains suggests changes in behaviors should be further investigated by testing the participation of postsynaptic mechanisms, measuring of cholinergic markers during earlier development periods and the possible participation of other neurotransmitter systems to clearly reveal the role of the cholinergic system following in utero MSF exposure. PMID- 17920113 TI - Pregnancy in the domestic cat after vaginal or transcervical insemination with fresh and frozen semen. AB - The objective was to compare pregnancy rates in domestic cats using fresh semen for intravaginal artificial insemination (IVI), either at the time of hCG treatment for induction of ovulation, or 28 h later, and to compare pregnancy rates following IVI or transcervical intrauterine insemination (IUI) of frozen thawed semen. Eighteen queens were inseminated during 39 estrus cycles. Fresh semen with 13.5+/-5.4 x 10(6) sperm (range, 6.8-22 x 10(6)) collected by electroejaculation from four male cats was used in Experiment 1, and cryopreserved semen (20 x 10(6) sperm, with 70+/-5% post-thaw motility) from one male cat was used in Experiment 2. Serum concentrations of estradiol-17beta and progesterone were determined in most queens on the day of AI and again 30-40 days later. Treatment with 100 IU of hCG 3 days after the onset of estrus induced ovulation in 95% of treated queens. Pregnancy rates to IVI with fresh semen at the time of hCG administration versus 28 h later were not different (P=0.58); overall 33% (5/15) of the queens became pregnant. For frozen-thawed semen, AI was consistently done 28h after hCG administration; IUI and IVI resulted in pregnancy rates of 41.7% (5/12), whereas no queen (0/12) became pregnant by IVI (P=0.0083). In conclusion, an acceptable pregnancy rate was obtained with frozen-thawed semen in the domestic cat by non-surgical transcervical IUI; this method might also be useful in other small felids. PMID- 17920115 TI - Axial shrinkage-stress depends upon both C-factor and composite mass. AB - OBJECTIVES: To measure and then mathematically model polymerization stress dependence upon systematic variations of C-factor (bonded/unbonded area ratio) for the Bioman instrument [1], recording stress by free cantilever-beam deflection; compliance 1.5 microm/MPa. METHODS: A light-cured resin-composite (RZD103; Ivoclar) with 57% (v/v) 450 nm filler was studied. Facing surfaces: glass slab and steel rod-end, constituting the Bioman test chamber, being perpendicular to the measured axial stress-direction, were varied: (a) with rod diameters (phi), from 1 to 10mm in 1mm increments (with 0.8mm gap height); and then (b) with gap heights (h) in 16 steps from 0.05 to 1.50mm (with phi=10mm). For each h and phi combination, giving C-factors ranging from 0.6 to 100, shrinkage-stress was recorded for 1h from start of 40s irradiation at 600 mW cm( 2) for photo-polymerization at 23 degrees C (n=3). Shrinkage-stress (S(sigma)) was plotted directly as functions of h, phi, and C and also per unit composite mass, (S(sigma)g(-1)). ANOVA and Tukey's statistics were applied. RESULTS: Series A-diameter variation; with C-factor increasing from 0.6 to 6, gave an exact exponential decrease in S(sigma) from 45 to 8 MPa. Series B-height variation; with C-factor increasing from 3 to 100, gave increasing S(sigma) from 1 to 8 MPa. Since composite mass played an equally dominant role, plots of stress-variations per unit composite mass, (S(sigma)g(-1)) separated these effects, confirming progressive off-axial stress-relief with increasing h. SIGNIFICANCE: (i) Values of h=0.8 and phi=10mm, recommended [1] for Bioman use, were confirmed as appropriate. Every lab instrument for measuring S(sigma) necessarily embodies specific C-factors and compliance values in the instrument design. (ii) Configuration (C) factor is recognized as an important parameter affecting manifestation of shrinkage-stress within restorative cavities and luting gaps. However, the restorative mass must equally be considered when translating shrinkage-science into specific clinical recommendations. PMID- 17920114 TI - HEMA reduces cell proliferation and induces apoptosis in vitro. AB - OBJECTIVES: Methacrylate monomers have been identified in aqueous extracts of freshly cured compomers. Both cells in the pulpal cavity and various cells of the oral mucosa can potentially be exposed to these leachables. Short-term exposure to dental monomers at relatively high concentrations induces adverse biological effects in vitro. The mechanisms involved have not been fully elucidated although involvement of various signaling pathways including ROS formation, activation of MAP-kinases and caspases has been suggested. The aim of this study was to investigate potential cellular responses following long-term exposure to relatively low and potentially more clinical relevant HEMA concentrations. METHODS: A submandibular gland cell line was exposed to HEMA (20-600 microM) for up to 72h. The impact on cell proliferation, apoptosis, and possible underlying mechanisms was assessed by flow cytometry, microscopy and western blotting. RESULTS: Exposure to HEMA (600 microM) resulted in reduced cell proliferation after 24h and increased apoptosis after 60h. Further, we observed ATM dependent phosphorylation of p53, advocating an initial DNA damage in the HEMA exposed cells. SIGNIFICANCE: In conclusion, we show that exposure to relatively low concentration of HEMA for a prolonged time result in cell death, possibly as a consequence of DNA damage. PMID- 17920116 TI - Tobacco smoking, harm reduction, and nicotine product regulation. PMID- 17920117 TI - Restoration of insulin sensitivity following treatment with imatinib mesylate (Gleevec) in non-diabetic patients with chronic myelogenic leukemia (CML). PMID- 17920118 TI - Comprehensive analysis of FOXP3 mRNA expression in leukemia and transformed cell lines. AB - Studies of FOXP3 expression have thus far focused on T cells, including both normal and malignant T cells. In particular, adult T cell leukemia/lymphoma (ATLL) cells have been studied intensively because their phenotype resembles that of normal CD4(+)CD25(+) regulatory T (Treg) cells. However, a comprehensive study of FOXP3 expression covering all hematopoietic cell lineages has not yet been performed. In this study, FOXP3 mRNA expression was examined by quantitative PCR using a large collection of human hematopoietic cell lines derived from leukemia/lymphoma or virus-transformation, including cells lines with T, B, plasmacytoid, myeloid, monocytic, megakaryocytic, erythroid, and NK lineages. Unexpectedly, we found FOXP3 mRNA expression in a number of cell lines belonging to all of the cell lineages investigated. In sharp contrast, FOXP3 protein expression was found in only three cell lines, all of which were HTLV-I-infected. Several non-T cell lines expressed higher levels of mRNA but were still negative for protein expression. The broad mRNA expression contrasts with the restricted protein expression of FOXP3 in human hematopoietic cell lines, suggesting that post-transcriptional control mechanisms may control FOXP3 protein expression. PMID- 17920119 TI - Erythrocytic nuclear abnormalities in wild and caged fish (Liza aurata) along an environmental mercury contamination gradient. AB - Laranjo basin (Aveiro, Portugal) has been subjected to mercury contamination from a chlor-alkali plant, presenting a well-described mercury gradient. This study aims the assessment of mercury genotoxicity in this area by measuring erythrocytic nuclear abnormalities (ENA) frequency in the mullet Liza aurata, and its relation with total mercury concentration (Hg(t)) in blood. Wild fish were seasonally analysed, and, complementarily, fish were caged for 3 days at three locations differing on their distances to the mercury source. The results from Laranjo were compared with those from a reference area (S. Jacinto). Wild fish from Laranjo showed elevated ENA frequency in summer and autumn in concomitance with increased blood Hg(t). Surprisingly, no ENA induction was found in winter, despite the highest blood Hg(t), which may be explained by haematological dynamics alterations, as supported by a decreased immature erythrocytes frequency. Caged fish displayed ENA induction only at the closest site to the contamination source, also showing a correlation with blood Hg(t). PMID- 17920120 TI - Cholera toxin transiently inhibits porcine T cell proliferation in vitro. AB - Cholera toxin (Ctx) is an important mucosal adjuvant with potential experimental applications in pigs. However, little is known about the direct effects of Ctx on porcine immune cells. Therefore, we analysed the influence of Ctx on mitogen induced lymphocyte proliferation. Ctx inhibited peripheral blood mononuclear cell (PBMC) proliferation with an IC50 of 34+/-17 ng/mL. This inhibition was not due to increased cell death. Lymphoblast formation in cultures stimulated with concanavalin A and Ctx was decreased at 24 h, but had reached the levels of control cultures again at 72 and 120 h, indicating that suppression was transient. Analysis of T cell subsets revealed that Ctx treatment specifically reduced the percentage of CD4-CD8+ and gammadelta T cells, whereas the proportion of CD4+CD8- increased. Furthermore, Ctx caused secretion of IL-10 by PBMC cultures, but depressed TNFalpha secretion. PMID- 17920121 TI - The process of evaluating a capacity-building support initiative for HIV positive South Africans. AB - This paper describes the process of evaluating a capacity-building initiative involving HIV positive South Africans. The evaluation was conducted within a participatory evaluation framework, which aims at empowering participants through social action. The rationale is that an understanding and acceptance of evaluation results promotes commitment to act. Forty four members of the Kudu Support Group and 23 non-members participated in the evaluation. Methods included in-depth interviews, a survey, repeated measures of the General Health Questionnaire-28 (GHQ-28), a SWOT analysis, interviews with health workers, field notes, and participatory observation. The capacity-building initiative resulted in empowerment through paid employment for support group members, the establishment of a vegetable garden, increased skills and efficacy, access to resources and networks, a perceived reduction of stigma, perceived improvement of social relations, and improved scores on the GHQ-28. Lessons that emerge out of this study are the importance of integrating evaluation as a part of ongoing activities throughout the research project; to ensure that methods and results are understandable and useful to for the participants; that the evaluator has an in-depth understanding of the project and context; and to introduce frequent and simple internal evaluation tools to guide efforts to improve community-based groups' functioning and activities. PMID- 17920122 TI - Mentally disordered offenders and the European Court of Human Rights. AB - Mentally disordered offenders find themselves at the intersection of the healthcare system and the criminal justice system in most European countries. Decisions on their care often involve lengthy discussions in relation to care versus control in society. In this article, the focus is on one aspect of this debate - that of human rights. An analysis of cases, taken to the European Court of Human Rights by mentally disordered offenders, demonstrates the difficulties inherent in ensuring appropriate care to individuals and safeguards to the public at the same time. The issues raised include the problems raised by indeterminate sentences, the use of detention for preventive purposes, and debates about treatment. The countries represented in this selection of cases are Belgium, Norway, Poland, the Netherlands, Russia and the United Kingdom. PMID- 17920123 TI - Chemopreventive agents induce programmed death-1-ligand 1 (PD-L1) surface expression in breast cancer cells and promote PD-L1-mediated T cell apoptosis. AB - Chemotherapy has been widely used in cancer treatment. However, the prognosis of the cancer patients following chemotherapy has not been substantially improved. Alternative strategies such as immunotherapy and their combinations with chemotherapy are now being considered. Yet, the effects of chemotherapy on the immune responses of cancer cells are not clear. Cancer immunoresistance and immune escape are major obstacles in immunotherapy. In the present studies, we examined the effects of chemopreventive agents, paclitaxel, etoposide and 5 fluorouracil, on the surface expression of programmed death-1-ligand 1 (PD-L1), a negative regulator of T cell anti-tumor immunity. Interaction of PD-L1 on cancer cells with programmed death receptor 1 (PD-1) on T cells has been reported to inhibit the proliferation of tumor-reactive cytotoxic T cells and induce T cell apoptosis, which could be an important mechanism in the development of cancer immunoresistance. We demonstrated that those chemopreventive agents were able to induce PD-L1 surface expression in human breast cancer cells, which then promoted PD-L1-mediated T cell apoptosis. Our studies reveal a potential link between chemotherapy and cancer immunoresistance. PMID- 17920124 TI - Interleukin-1beta gene in orange-spotted grouper, Epinephelus coioides: molecular cloning, expression, biological activities and signal transduction. AB - IL-1beta, a key mediator of inflammation, orchestrates a variety of immune responses by initiating gene expression. Herein, we have cloned and sequenced the IL-1beta in orange-spotted grouper (Epinephelus coioides), produced soluble mature recombinant IL-1beta in Escherichia coli, and characterized its biological properties and downstream signal transduction. The grouper IL-1beta cDNA was 1364bp in length, containing an open reading frame of 765bp. The predicted protein of 254 amino acids revealed the presence of the IL-1 family signature motif and the absence of a conventional ICE cut site. Phylogenetically, the grouper IL-1beta clustered closely with those of teleost belonging to Perciformes and apart from those of mammals. The grouper IL-1beta was constitutively expressed in almost all tissues examined, and was augmented in PBL after the addition of LPS or Poly I:C in vitro. The prokaryotically produced rIL-1beta significantly stimulated the proliferation of grouper head kidney cells, and activated gene expression of IL-1beta and COX-2. Moreover, the rIL-1beta-induced IL-1beta and COX-2 expression were reduced by p38 MAPK inhibitor (SB203580) and JNK inhibitor (SP600125), respectively. Taken together, the present study indicated that grouper IL-1beta may have an important role in grouper immune system and activate similar downstream cascades as its mammalian counterparts. PMID- 17920125 TI - The warm temperature acclimation protein Wap65 as an immune response gene: its duplicates are differentially regulated by temperature and bacterial infections. AB - The warm temperature acclimation related 65kDa protein (Wap65) in teleost fish shares high structural similarities with mammalian hemopexins. Recent studies using microarray analysis indicated that this temperature acclimation protein may also be involved in immune responses. To provide evidence of its potential involvement in immune responses after bacterial infections, we have identified and characterized two types of Wap65 genes in channel catfish, referred to as Wap65-1 and Wap65-2, respectively. While Wap65-1 and Wap65-2 are both structurally similar to the mammalian hemopexins, they exhibit highly differential patterns of spatial expression. Wap65-1 was expressed in a wide range of tissues, whereas Wap65-2 was only expressed in the liver. Their regulation with warm temperature and bacterial infections was also highly different: Wap65-1 was constitutively expressed, whereas Wap65-2 was highly regulated by both warm temperature and bacterial infections, and warm temperature and bacterial infections appeared to synergistically induce the expression of Wap65-2. The great contrast of expression patterns and regulation of the two catfish Wap65 genes suggested both neofunctionalization and partitioning of their functions. Phylogenetic analysis indicated that the duplicated catfish Wap65 genes were evolved not only from whole genome duplication, but also from tandem, intrachromosomal gene duplications. Taken together, the results of this study suggest that Wap65 genes are not only important for its classical role as a warm temperature acclimation protein, but more importantly, may also function as an immune response protein. PMID- 17920126 TI - Rainbow trout suppressor of cytokine signalling (SOCS)-1, 2 and 3: molecular identification, expression and modulation. AB - Suppressor of cytokine signalling (SOCS) proteins are inhibitors of cytokine signalling pathways. Three SOCS genes, SOCS-1, 2 and 3, have been identified and their sequences analyzed in an economically important fish, rainbow trout (Oncorhynchus mykiss, Walbaum). In general, these three SOCS molecules are well conserved especially in the SRC homology 2 and the SOCS domains, with sequence identities between trout and mammals ranging from 41 to 42, 50 to 51, and 58 to 61% for SOCS-1, 2 and 3, respectively. The identities within fish species are slightly higher, with sequence identities between trout and the other fish species at 44-46, 64-70, and 71-76% for SOCS-1, 2 and 3, respectively. All the SOCS-1, as well as all the SOCS-2 or 3 molecules from different species are grouped together in phylogenetic tree analysis with high bootstrap support, with the fish molecules in each type grouping closely together. The expression of the trout SOCS-1, 2 and 3 genes are detectable by real-time PCR in all the eight tissues studied; the gills, skin, muscle, liver, spleen, head kidney, intestine and brain. SOCS-1 is highly expressed in intestine, head kidney, spleen, gills and skin. SOCS-2 is highly expressed in brain, head kidney, muscle, spleen, gills, skin and intestine. The expression of SOCS-3 is the highest among the three SOCS genes in all the tissues except in intestine, brain and liver. The modulation of SOCS gene expression is shown to be cytokine and cell type dependent. While interferon-gamma up-regulates the expression of all the three SOCS genes in both the fibroid RTG-2 and the monocyte/macrophage RTS-11 cell lines, interleukin-1beta only up-regulates SOCS gene expression in the RTG-2 cell line, with little, if any, effect in the RTS-11 cell line. PMID- 17920127 TI - Methylmercury activates ASK1/JNK signaling pathways, leading to apoptosis due to both mitochondria- and endoplasmic reticulum (ER)-generated processes in myogenic cell lines. AB - Cellular stress responses following exposure to methylmercury (MeHg) were investigated using myogenic cell lines that showed different susceptibilities to MeHg. The susceptible cell line showed apoptosis within 24h after exposure to low levels of MeHg. The activation of caspase 12, 9, and 3 was detected in the apoptotic cells at 14-16 h after MeHg exposure, suggesting that MeHg causes apoptosis via both mitochondria- and endoplasmic reticulum (ER)-generated processes. An early increase in the level of intracellular reactive oxygen species (ROS) was quantitatively recognized since 2-3h after exposure to MeHg in both MeHg-susceptible and non-susceptible cell lines; however, the increase was lower in the latter cell line. The phosphorylation of apoptosis signal-regulating kinase 1 (ASK1) was also recognized in both cell lines, with the increase in intracellular ROS. However, the activation of stress-activated protein kinase/c Jun N-terminal kinase (SAPK/JNK) pathways was observed only in the MeHg susceptible cell line. In contrast, the non-susceptible cell line exhibited activation of the cell survival ERK pathway. Up-regulation of metallothioneine I and Hic-5 mRNAs encoding proteins induced by oxidative stress was recognized during the early stage of MeHg cytotoxicity in the MeHg-susceptible cell line. Quantitative real-time PCR and western blot analyses confirmed that ER stress is a late event during MeHg cytotoxicity. Coaddition of the antioxidant Trolox dramatically suppressed the increase in the level of ROS, activation of caspases and, finally, apoptosis. However, later treatment with Trolox attenuated its protective effect against MeHg cytotoxicity. The results indicate that failure to protect cells against the early oxidative stress triggers ER stress and apoptosis processes. Combined treatment with protective factors against oxidative and ER stresses is necessary, especially in the later stages of MeHg cytotoxicity. PMID- 17920128 TI - Modulation of cholinergic systems by manganese. AB - Information on changes in the central nervous system (CNS) cholinergic systems following exposure to manganese are considerably less extensive than that associated with other neurotransmitter systems. However, experimental and clinical evidence support the notion that cholinergic activity plays a key role in the pathophysiology of manganese-induced neurotoxicity. Manganese acts as a chemical stressor in cholinergic neurons in a region-specific manner causing breakdown of the cellular homeostatic mechanisms. In fact, a number of cholinergic synaptic mechanisms are putative targets for manganese activity: presynaptic choline uptake, quantal release of acetylcholine into the synaptic cleft, postsynaptic binding of acetylcholine to receptors and its synaptic degradation by acetylcholinesterase. Moreover, manganese significantly influences astrocytic choline transport systems and astrocytic acetylcholine-binding proteins. Thus, manganese exerts its effect on the highly dynamic reciprocal relationship between astrocytes and cholinergic neurons. Cholinergic afferents are crucial in the physiology of locomotion, cognition, emotion and behavioral response, and therefore, it is not surprising that the anatomical selectivity of most manganese-induced cholinergic effects is compatible with the clinical correlates of manganism, which involves impairment of emotional response, decline in higher cortical functions and movement disorder. Manganism, also referred to as Parkinson's-like disorder, is initially manifested by a neuropsychiatric syndrome (locura manganica), the most frequent symptoms and signs of which are compulsive behavior, emotional lability, visual hallucinations and flight of ideas, cognitive decline and memory loss. These signs and symptoms are followed by an extrapyramidal syndrome, which shares numerous clinical and pathophysiological characteristics with idiopathic Parkinson's disease (PD). This natural history of disease could be a clinical reflection of the preferential involvement of the cholinergic systems, initially in the septo-hippocampus and later in the basal ganglia. These observations highlight the importance of studying the role of the CNS cholinergic systems in manganese-mediated neurotoxicity. PMID- 17920132 TI - Action-based sensory encoding in spinal sensorimotor circuits. AB - The concept of a modular organisation of the spinal withdrawal reflex circuits has proven to be fundamental for the understanding of how the spinal cord is organised and how the sensorimotor circuits translate sensory information into adequate movement corrections. Recent studies indicate that a task-related body representation is engraved at the network level through learning-dependent mechanisms involving an active probing procedure termed 'somatosensory imprinting' during development. It was found that somatosensory imprinting depends on the tactile input that is associated with spontaneous movements that occur during sleep and results in elimination of erroneous connections and establishment of correct connections. In parallel studies it was found that the strength of the first order tactile synapses in rostrocaudally elongated zones in the adult dorsal horn in the lower lumbar cord is related to the modular organisation of the withdrawal reflexes. Hence, the topographical organisation of the tactile input to this spinal area seems to be action-based rather than a simple body map as previously thought. Far from being innate and adult like at birth, the adult organisation seems to emerge from an initial 'floating' and diffuse body representation with many inappropriate connections through profound activity-dependent rearrangements of afferent synaptic connections. It is suggested that somatosensory imprinting plays a key role in the self-organisation of the spinal cord during development. PMID- 17920131 TI - Spontaneous rhythmic activity in early chick spinal cord influences distinct motor axon pathfinding decisions. AB - During embryonic development chick and mouse spinal cords are activated by highly rhythmic episodes of spontaneous bursting activity at very early stages, while motoneurons are still migrating and beginning to extend their axons to the base of the limb. While such spontaneous activity has been shown to be important in refining neural projections once axons have reached their targets, early pathfinding events have been thought to be activity independent. However, in-ovo pharmacological manipulation of the transmitter systems that drive such early activity has shown that early motor axon pathfinding events are highly dependent on the normal pattern of bursting activity. A modest decrease in episode frequency resulted in dorsal-ventral pathfinding errors by lumbar motoneurons, and in the downregulation of several molecules required to successfully execute this guidance decision. In contrast, increasing the episode frequency was without effect on dorsal-ventral pathfinding. However, it prevented the subsequent motoneuron pool specific fasciculation of axons and their targeting to appropriate muscles, resulting in marked segmental pathfinding errors. These observations emphasize the need to better evaluate how such early spontaneous electrical activity may influence the molecular and transcription factor pathways that have been shown to regulate the differentiation of motor and interneuron phenotypes and the formation of spinal cord circuits. The intracellular signaling pathways by which episode frequency affects motor axon pathfinding must now be elucidated and it will be important to more precisely characterize the patterns with which specific subsets of motor and inter-neurons are activated normally and under conditions that alter spinal circuit formation. PMID- 17920134 TI - Brain-computer interfaces (BCIs): detection instead of classification. AB - Many studies over the past two decades have shown that people can use brain signals to convey their intent to a computer through brain-computer interfaces (BCIs). These devices operate by recording signals from the brain and translating these signals into device commands. They can be used by people who are severely paralyzed to communicate without any use of muscle activity. One of the major impediments in translating this novel technology into clinical applications is the current requirement for preliminary analyses to identify the brain signal features best suited for communication. This paper introduces and validates signal detection, which does not require such analysis procedures, as a new concept in BCI signal processing. This detection concept is realized with Gaussian mixture models (GMMs) that are used to model resting brain activity so that any change in relevant brain signals can be detected. It is implemented in a package called SIGFRIED (SIGnal modeling For Real-time Identification and Event Detection). The results indicate that SIGFRIED produces results that are within the range of those achieved using a common analysis strategy that requires preliminary identification of signal features. They indicate that such laborious analysis procedures could be replaced by merely recording brain signals during rest. In summary, this paper demonstrates how SIGFRIED could be used to overcome one of the present impediments to translation of laboratory BCI demonstrations into clinically practical applications. PMID- 17920133 TI - Morphological and physiological properties of serotonergic neurons in dissociated cultures from the postnatal rat dorsal raphe nucleus. AB - We have developed dissociated primary cultures of the dorsal raphe nucleus from postnatal 9-12-day-old rats. The nucleus was dissected out from brain slices, dissociated, and cultured over a glial feeder layer. Serotonin immunocytochemistry revealed that 62% of cultured neurons were serotonergic. There was no significant difference in diameters between serotonergic and non serotonergic neurons. With the whole-cell patch-clamp method, cultured neurons were tested for responses to 8-hydroxydipropylaminotetraline (8-OH-DPAT, a selective agonist for 5-HT(1A)), and then treated with serotonin immunocytochemistry. Ninety-two percent of neurons responding to 8-OH-DPAT were serotonergic. These results were used to identify serotonergic neurons. In most cases, serotonergic neurons did not show spontaneous firings of action potentials. Constant current depolarizations elicited trains of action potentials that usually did not show marked adaptation. Application of 8-OH-DPAT inhibited action potential firing. The current-voltage relation of the 8-OH-DPAT-induced current indicated an inward rectification with its reversal potential near E(K). Serotonergic neurons were depolarized by phenylephrine, bombesin, and gastrin releasing peptide. This culture system will serve as a useful tool for elucidating the cellular, physiological, and molecular properties of brain serotonergic neurons. PMID- 17920135 TI - Central visfatin causes orexigenic effects in chicks. AB - Intracerebroventricular injection of visfatin caused increased feed intake and pecking efficiency, but did not affect water intake in chicks. Visfatin-treated chicks had increased c-Fos immunoreactivity in the lateral hypothalamus, decreased reactivity in the ventromedial hypothalamus and the dorsomedial hypothalamus, infundibular nucleus, periventricular nucleus, paraventricular nucleus were not affected. A low dose of visfatin increased locomotion. We conclude that intracerebroventricular injection of visfatin causes orexigenic effects in chicks. PMID- 17920136 TI - Gene inactivation confirms the identity of enzymes involved in nematode phosphorylcholine-N-glycan synthesis. AB - An unusual feature of nematodes is the covalent attachment of immunomodulatory phosphorylcholine (PC) moieties to N-type glycans. Our previous work on the filarial nematode glycoprotein ES-62 has enabled us to predict the identity of enzymes necessary for PC-N-glycan biosynthesis. Here, we addressed these predictions using gene knockout technology applied to C. elegans and present two pieces of confirmatory data. Employing a triple null mutant worm lacking all three genes that encode active UDP-N-acetyl-D-glucosamine: alpha-3-D-mannoside beta1, 2-N-acetylglucosaminyltransferase I (GnT I) we have confirmed our earlier prediction that a crucial step in the generation of the substrate for PC transfer is addition of terminal GlcNAc to the alpha1-3-linked mannose residue of the glycan by GnT I. Second, by silencing genes responsible for expressing enzymes of the Kennedy pathway of phosphatidylcholine biosynthesis by RNA interference (RNAi), we have confirmed our belief for a role for diacylglycerol: choline phosphotransferase (CPT) in PC-N-glycan biosynthesis. PMID- 17920137 TI - The F-box protein CFB2 is required for cytokinesis of bloodstream-form Trypanosoma brucei. AB - F-box proteins serve as mediators in targeting bound target proteins for ubiquitination and destruction. We here describe the roles of two F-box proteins, CFB1 and CFB2, in the trypanosome cell cycle. Five almost identical copies of CFB1 are arranged in a direct tandem repeat on Trypanosoma brucei chromosome 1; immediately downstream is a single CFB2 gene. RNAi targeting CFB1 in bloodstream form trypanosomes had a transient effect on growth and mitosis. Depletion of CFB2, in contrast, resulted in immediate growth arrest and rapid cell death. CFB2 depleted cells accumulated nuclei and kinetoplasts with the corresponding numbers of basal bodies and flagella. The CFB2 transcript was less abundant in procyclic form trypanosomes, and RNAi against CFB2 in these forms had no effect on growth. These results suggest that CFB2 is required for bloodstream-form trypanosome cytokinesis. PMID- 17920138 TI - Cystatin B and its EPM1 mutants are polymeric and aggregate prone in vivo. AB - Progressive myoclonus epilepsy type 1 (EPM1) is a neurodegenerative disease correlating with mutations of the cystatin B gene. Cystatin B is described as a monomeric protein with antiprotease function. This work shows that, in vivo, cystatin B has a polymeric structure, highly resistant to SDS, urea, boiling and sensitive to reducing agents and alkaline pH. Hydrogen peroxide increases the polymeric structure of the protein. Mass spectrometry analysis shows that the only component of the polymers is cystatin B. EPM1 mutants of cystatin B transfected in cultured cells are also polymeric. The banding pattern generated by a cysteine-minus mutant is different from that of the wild-type protein as it contains only monomers, dimers and some very high MW bands while misses components of MW intermediate between 25 and 250 kDa. Overexpression of wild-type or EPM1 mutants of cystatin B in neuroblastoma cells generates cytoplasmic aggregates. The cysteine-minus mutant is less prone to the formation of inclusion bodies. We conclude that cystatin B in vivo has a polymeric structure sensitive to the redox environment and that overexpression of the protein generates aggregates. This work describes a protein with a physiological role characterized by highly stable polymers prone to aggregate formation in vivo. PMID- 17920139 TI - A case report of myocardial infarction in young patient with a parental history of premature cardiovascular death: combination of prothrombotic gene mutations. AB - We report a case of myocardial infarction at a young age in a subject heterozygous for the G20210A prothrombin gene variant and homozygous for the C677T MTHFR polymorphism, who presented a strong family history of atherothrombosis. Genetic screening for inherited thrombophilia, especially in the presence of a strong familiarity, may be a critical information for secondary prevention of arterial thrombosis. PMID- 17920140 TI - Cardiology in contemporary China: an update. PMID- 17920141 TI - Patent foramen ovale and ascending aortic aneurysm with platypnea-orthodeoxia syndrome. AB - Platypnea-orthodeoxia is a rare syndrome characterized by dyspnea induced by the upright position and relieved by supine position and an arterial deoxygenation increased by the upright position which improves during recumbency. Several anatomical factors that can alter the atrial anatomy and facilitate shunting through an interatrial defect have been related with this syndrome. In many cases, this syndrome has been associated with patent foramen ovale and right-to left shunt. Rarely platypnea-orthodeoxia syndrome has been described associated with an aortic aneurysm too. We present a case of platypnea-orthodeoxia syndrome in a 76-year-old Italian man with patent foramen ovale and ascending aortic aneurysm. PMID- 17920142 TI - High resolution imaging by organic secondary ion mass spectrometry. AB - Secondary-ion mass spectrometry (SIMS) is based on the acceleration of high energy primary ions onto a target. Secondary electrons, neutrals and ions are emitted from the target, reflecting its chemical composition. This enables simultaneous analysis and localization of target molecules, giving valuable information that is difficult or impossible to obtain with other analytical methods. The secondary ions can be extracted and detected by any type of mass analyzer. SIMS is unique in its ability to detect several target molecules simultaneously in small samples and to image their localization at subcellular resolution. The recent development of bioimaging SIMS opens up new possibilities in biotechnology and biological research with applications in biomedicine and pathology. The current development of this technique has the potential to become as important for biotechnology as the advent of the electron microscope, confocal microscope or in situ hybridization. PMID- 17920143 TI - FDG-PET scan and treatment planning for early stage Hodgkin lymphoma. PMID- 17920144 TI - Affective modulation of eyeblink reactions to noxious sural nerve stimulation: a supraspinal measure of nociceptive reactivity? AB - Research suggests affective picture-viewing modulates subjective and physiological reactions to noxious stimulation (pain report, heart rate acceleration, skin conductance response, nociceptive flexion reflex). Because the nociceptive flexion reflex (a spinal reflex) is modulated by picture-viewing, this suggests affective processes are able to modulate afferent nociception at spinal levels. This highlights the importance of assessing nociceptive reactivity from physiological measures mediated at different levels of the neuraxis (spinal vs. supraspinal) to help elucidate the mechanisms associated with pain regulation. The present study examined whether affective pictures modulate eyeblink reactions (a supraspinal reflex) to noxious stimulation. Healthy men and women (N=23) were recruited from the psychology subject pool to participate. Pictures (attack, loss, neutral, food, erotica) that manipulated affective valence and arousal were presented and noxious electrodermal stimulations were delivered to the sural nerve. Picture duration (500-ms vs. 6-s) was also manipulated, balanced across picture content. Results suggested affective valence and arousal contributed to the modulation of eyeblinks. Eyeblinks were larger during highly arousing unpleasant pictures (attack) than highly arousing pleasant pictures (erotica), but low arousal pictures (loss, food) did not lead to significant modulation. Affective modulation was independent of picture duration or the perceived painfulness of noxious stimulation. This study suggests eyeblink reactions can serve as a supraspinal outcome in procedures used to study affective modulation of pain and nociception. PMID- 17920145 TI - The effect of mechanical context on attentional cost in unimanual coordination. AB - The purpose of this study was to address the attentional cost of sensorimotor coordination by determining if changes to the mechanical context of movement would influence the ability to attend and respond to an alternate stimulus. Nine right-handed participants performed rhythmic pronation and supination movements of the forearm in time with an auditory metronome. A secondary task, consisting of a pedal response to visual probe stimuli, was employed to infer the attentional cost of the coordination task. When the axis-of-rotation (AOR) was placed below the long axis of the forearm, the average time to react (RT) to the probe stimuli was greater for the supinate-on-the-beat condition than for the pronate-on-the-beat condition. Conversely, with the AOR above the forearm, RT for the pronate-on-the-beat pattern was greater than that for the supinate-on-the beat pattern. Thus, changing the mechanical context of an upper limb coordination task altered the central processing cost required to maintain pattern stability. This finding provides further evidence that the attentional resources required to produce a particular movement are determined by the ease with which the action is executed by the sensorimotor system. PMID- 17920146 TI - Role of arms in somersaulting from compliant surfaces: a simulation study of springboard standing dives. AB - The role of arms in compliant-surface jumping for maximizing backward somersault rotations is studied using multi-segment models and is applied to springboard diving. The surface (springboard) is modeled by a rigid bar with a rotational spring with a hinged end and point mass at the tip. Planar four- and five-segment human models are used (with the fifth segment representing the arms) and are driven by torque actuators at the ankle, knee, hip, and shoulder. Each joint torque is the product of maximum isometric torque and three variable functions depending on instantaneous joint angle, angular velocity, and activation level, respectively. Movement simulation starts from a balanced initial posture and ends at jump takeoff. The objective is to find joint torque activation patterns during board contact so that the number of backward rotations in flight is maximized. Kinematic differences in jumps with and without arms are mainly in smaller takeoff vertical velocity and more flexed knee and hip in the former. In both jumps, joint torque/activations are similar in their minor flexion-full extension patterns. Maximum hip torque is larger with arms but maximum knee torque is larger without arms. Except at the knee, more joint work can be done with arm swing. Total angular momentum is increased considerably by arm motion because of its remote contribution. Consequently segment remote contributions to total angular momentum are much larger in jumping with arms. Shoulder strength helps generate angular momentum only to a certain limit. If more work is used to generate horizontal velocity away from the board, the amount of total angular momentum is reduced. PMID- 17920147 TI - Distinct properties of corticothalamic and primary sensory synapses to thalamic neurons. AB - An unique synaptic feature of thalamic relay neurons is that, in addition to receiving primary sensory synapses, they receive massive feedback synapses originating from the cerebral cortex (corticothalamic synapses). These two synapses are both glutamatergic and coordinate the firing responses of thalamic neurons. It has been revealed in the past decade that various glutamate receptors are involved in synaptic responses in the thalamus. However, differences in the compositions of glutamate receptors between corticothalamic and primary sensory synapses have not been fully determined. This update article aims to provide an overview of the differences in the synaptic properties, including the compositions of glutamate receptors, between primary sensory and corticothalamic synapses in the ventrobasal nucleus of the somatosensory thalamus, and then elucidate how these differences in the two synapses influence the firing properties of thalamic neurons. PMID- 17920149 TI - Adenosine induces expression of glial cell line-derived neurotrophic factor (GDNF) in primary rat astrocytes. AB - Adenosine, which accumulates rapidly during ischemia due to the breakdown of ATP, has beneficial effects in many tissues. We examined whether adenosine induces the production of glial cell line-derived neurotrophic factor (GDNF) in cultured astrocytes. We evaluated GDNF mRNA expression and GDNF production in astrocytes cultured with adenosine and the adenosine selective receptor agonists 5-(N ethylcarboxamido) adenosine (NECA), N(6)-cyclopentyladenosine (CPA) and 2-p-(2 carboxyethyl) phenethylamino-5'-N-ethylcarboxamindo-adenosine hydrochloride (CGS 21680). Moreover, we examined the possibility that the expression of GDNF is regulated differently in cultured astrocytes from the stroke-prone spontaneously hypertensive rat (SHRSP) than in those from Wistar Kyoto rats (WKY). In this study, we confirmed that adenosine and the selective A(2B) adenosine receptor agonist NECA induced the expression of GDNF in cultured astrocytes. The A(2B) receptor antagonist alloxazine was able to inhibit the increase in extracellular GDNF produced by adenosine. Furthermore, the amounts of GDNF produced were significantly reduced in astrocytes of the adenosine-treated SHRSP compared with those of WKY. These results indicate that adenosine induces the expression of GDNF, and adenosine A(2B) receptors participate in the regulation of GDNF levels in astrocytes. This expression was attenuated in astrocytes of SHRSP compared with those of WKY. PMID- 17920148 TI - Inhibition of glial cell activation ameliorates the severity of experimental autoimmune encephalomyelitis. AB - Activated microglia and astrocytes have been implicated in the course of multiple sclerosis (MS) and its animal model: experimental autoimmune encephalomyelitis (EAE). MW01-5-188WH is a novel drug that selectively inhibits glial activation in the central nervous system (CNS). We report here that MW01-5-188WH is effective to ameliorate the severity of myelin oligodendrocyte glycoprotein (MOG)-induced EAE. Daily oral administration of MW01-5-188WH at 5mg/kg body weight reduced the clinical scores of EAE mice while having no influence on the disease incidence or animal mortality. Pathological examination revealed reduced numbers of microglia and astrocytes in the spinal cord of MW01-5-188WH-treated EAE mice. Moreover, MW01-5-188WH suppressed the release of key chemokines, which are involved in MS pathology, from cultured microglia and astrocytes. Taken together, our results indicate that treatments that suppress the activation of microglia and astrocytes should be pursued in future research for their potential as avenues for the treatment of MS. PMID- 17920150 TI - Analysis of proteins responsive to acetic acid in Acetobacter: molecular mechanisms conferring acetic acid resistance in acetic acid bacteria. AB - Acetic acid bacteria are used for industrial vinegar production because of their remarkable ability to oxidize ethanol and high resistance to acetic acid. Although several molecular machineries responsible for acetic acid resistance in acetic acid bacteria have been reported, the entire mechanism that confers acetic acid resistance has not been completely understood. One of the promising methods to elucidate the entire mechanism is global analysis of proteins responsive to acetic acid by two-dimensional gel electrophoresis. Recently, two proteins whose production was greatly enhanced by acetic acid in Acetobacter aceti were identified to be aconitase and a putative ABC-transporter, respectively; furthermore, overexpression or disruption of the genes encoding these proteins affected acetic acid resistance in A. aceti, indicating that these proteins are involved in acetic acid resistance. Overexpression of each gene increased acetic acid resistance in Acetobacter, which resulted in an improvement in the productivity of acetic acid fermentation. Taken together, the results of the proteomic analysis and those of previous studies indicate that acetic acid resistance in acetic acid bacteria is conferred by several mechanisms. These findings also provide a clue to breed a strain having high resistance to acetic acid for vinegar fermentation. PMID- 17920151 TI - Relationship between nitrate/nitrite reductase activities in meat associated staphylococci and nitrosylmyoglobin formation in a cured meat model system. AB - Quantitative determination of catalase, nitrate reductase, nitrite reductase and nitric oxide synthase activities (NOS) was performed on 11 different bacterial strains, mainly staphylococci, isolated from fermented sausages, bacon brine or cured meat products. All except one strain possessed catalase activity in the range from 1.0 to 6.1 micromol min(-1) ml(-1). Ten out of 11 bacteria strains showed nitrate reductase activity in the range between 50 and 796 nmol min(-1) ml(-1) and nine showed nitrite reductase activity in the range between 6 and 42 nmol min(-1) ml(-1). No evidence of NOS activity of the selected strains was detected. In a colour formation assay containing myoglobin all strains affected nitrosylmyoglobin (MbFe(II)NO) formation in assays containing nitrite, whereas only strains having nitrate reductase activity generated MbFe(II)NO in assays containing nitrate as the sole nitrosylating agent. The quantitative nitrate and nitrite reductase activity did not fully explain or correlate well with the observed rate of formation of MbFe(II)NO, which seemed to be more affected by the growth rate of the different strains. The mechanism of the reduction of nitrite into NO of strains not having nitrite reductase activity remains to be fully elucidated, but could be due to a dual-mode action of nitrate reductase capable of acting on nitrate. PMID- 17920152 TI - Does evidence exist for the presence of UDP-D-xylose:proteoglycan core protein beta-D-xylosyltransferase I orthologs in Pseudomonas fluorescens? PMID- 17920153 TI - Distinct human prolactin (hPRL) and growth hormone (hGH) behavior under bacteriophage lambda PL promoter control: temperature plays a major role in protein yields. AB - When producing recombinant protein for therapy, it is desirable not only to obtain substantial amounts of the protein, but also to make sure that potential contaminants such as inducing agents are not present in the final product. To prevent this, one can use expression systems in which the promoter (lambdaP(L)) is activated by a temperature shift that denatures a repressor (e.g., cIts). In this manner, hGH was successfully expressed and secreted in Escherichia coli periplasm, with specific yields well above 1 microg ml(-1) A(600)(-1), after a temperature shift from 30 to 42 degrees C. However, attempts to express a related hormone, human prolactin, employing the same protocol were unsuccessful, providing 0.03 microg ml(-1) A(600)(-1) at the most. A process is described in which this labile protein is obtained from a cIts(-) strain under optimized temperature condition (37 degrees C). The highest periplasmic secretions of prolactin ever reported were thus obtained: 0.92+/-0.10 microg ml(-1) A(600)(-1) at an optical density of approximately 3 A(600) units in shake flask cultures and approximately 1 microg ml(-1) A(600)(-1), at an OD of 35 A(600) units, via a rapid and flexible batch feed process in laboratory bioreactor. Purified hPRL was monomeric, correctly processed (Mr=22,906), properly folded and bioactive (51.5+/ 24.1 IU mg(-1)). PMID- 17920154 TI - Temperature-sensitive hydrogels with SiO2-Au nanoshells for controlled drug delivery. AB - Silica-gold (SiO(2)-Au) nanoshells are a new class of nanoparticles that consist of a silica dielectric core that is surrounded by a gold shell. These nanoshells are unique because their peak extinctions are very easily tunable over a wide range of wavelengths particularly in the near infrared (IR) region of the spectrum. Light in this region is transmitted through tissue with relatively little attenuation due to absorption. In addition, irradiation of SiO(2)-Au nanoshells at their peak extinction coefficient results in the conversion of light to heat energy that produces a local rise in temperature. Thus, to develop a photothermal modulated drug delivery system, we have fabricated nanoshell composite hydrogels in which SiO(2)-Au nanoshells of varying concentrations have been embedded within temperature-sensitive hydrogels, for the purpose of initiating a temperature change with light. N-isopropylacrylamide-co-acrylamide (NIPAAm-co-AAm) hydrogels are temperature-sensitive hydrogels that were fabricated to exhibit a lower critical solution temperature (LCST) slightly above body temperature. The resulting composite hydrogels had the extinction spectrum of the SiO(2)-Au nanoshells in which the hydrogels collapsed reversibly in response to temperature (50 degrees C) and laser irradiation. The degree of collapse of the hydrogels was controlled by the laser fluence as well as the concentration of SiO(2)-Au nanoshells. Modulated drug delivery profiles for methylene blue, insulin, and lysozyme were achieved by irradiation of the drug loaded nanoshell-composite hydrogels, which showed that drug release was dependent upon the molecular weight of the therapeutic molecule. PMID- 17920155 TI - Micromechanical control of cell and tissue development: implications for tissue engineering. AB - Tissue engineering approaches for repair of diseased or lost organs will require the development of new biomaterials that guide cell behavior and seamlessly integrate with living tissues. Previous approaches to engineer artificial tissues have focused largely on optimization of scaffold polymer chemistry and selection of appropriate biochemical additives (e.g., growth factors, adhesive ligands) to provide effective developmental control. However, recent work has shown that micromechanical forces and local variations of extracellular matrix (ECM) elasticity at the microscale regulate cell and tissue development both in vitro and in vivo. The micromechanical properties of the host tissue microenvironment also play a critical role in control of stem cell lineage switching. Here we discuss how new understanding of the fundamental role that mechanical forces play in tissue development might be leveraged to facilitate the development of new types of biomimetic materials for regenerative medicine, with a focus on the design of injectable materials that can target to injury sites, recruit stem cells and direct cellular self-assembly to regenerate functional tissues and organs in situ. PMID- 17920156 TI - Erlotinib, docetaxel, and gefitinib in sequential cohorts with relapsed non-small cell lung cancer. AB - BACKGROUND: Both docetaxel and erlotinib improve overall survival over best supportive care in non-small cell lung cancer (NSCLC). We assessed the effectiveness of erlotinib (E) and gefitinib (G) in patients with relapsed NSCLC in both second- and third-line settings, and compared this with that of docetaxel (D), in our clinical practice. METHODS: Sequential cohorts of patients with relapsed advanced stage NSCLC who had been treated with erlotinib (150 mg), gefitinib (250 mg), or docetaxel (75 mg/m(2)) were retrospectively identified from our database. The primary endpoint was overall survival. Secondary endpoints were response rate and progression-free survival. RESULTS: After adjusting for covariates, there was no significant difference in overall survival between the three drugs in both second-line (median E=24; G=25; D=43 weeks, p=0.17), and third-line (median E=31; G=24; D=29 weeks, p=0.61) settings. Response rates were also not statistically significant between the three drugs across both lines of treatment. CONCLUSIONS: Erlotinib, gefitinib, and docetaxel have similar effectiveness in this non-trial setting. PMID- 17920157 TI - Changes in plasmalemma K+Mg2+ -ATPase dephosphorylating activity and H+ transport in relation to seasonal growth and freezing tolerance of Festuca pratensis Huds. AB - Changes in plasmalemma K(+)Mg(2+)-ATPase dephosphorylating activity and H(+) transport were examined in freezing-tolerant and non-tolerant genotypes of the perennial grass species Festuca pratensis Huds. Enzyme activity and DeltamuH(+) were measured in plasmalemma fractions isolated from basal nodes and roots. Three types of experiments were undertaken: (i) a field experiment, utilizing the seasonal growth and cessation cycle of a perennial plant; (ii) a cold acclimation experiment in hydroponics; and (iii) an instant freezing test. A specific fluctuation in K(+)Mg(2+)-ATPase activity was found throughout the seasonal growth of the plants (i). The K(+)Mg(2+)-ATPase activity peaks for both the basal node and the root plasmalemma were determined early in the spring before the renewal of growth. The lowest activity values in roots occurred at the time approaching flowering, and in basal nodes at the transition into the growth cessation. The K(+)Mg(2+)-ATPase activity was approximately 50% lower in the basal node plasmalemma of freezing-tolerant plants than of non-tolerant ones, when assessed at the optimal growth stage in hydroponics. In hydroponics (ii) and in the freezing test (iii), temperature stress was followed by a more pronounced change in the level of K(+)Mg(2+)-ATPase activity than in that of H(+) transport, and this change was more clearly differentiated in the basal node plasmalemma of contrasting genotypes than in the roots. Stress response was manifested differently in freezing-tolerant and non-tolerant plants at cold acclimation (4-2 degrees C) and at freezing (-8 degrees C) temperatures. Proton transport regulation via coupled changes in the hydrolysed ATP/transported proton ratio, as an attribute of freezing-tolerant plants, is discussed. PMID- 17920158 TI - Protective action of salicylic acid against bean yellow mosaic virus infection in Vicia faba leaves. AB - In this study, morphological, ultrastructural and physiological modifications of faba bean (Vicia faba cv Giza 461) leaves in response to bean yellow mosaic virus (BYMV) infection and salicylic acid (SA) treatments were examined. Under BYMV stress, leaves showed symptoms including severe mosaic, mottling, crinkling, size reduction and deformations. Three weeks after virus inoculation, photosynthetic rate, pigment contents and transpiration rate were significantly reduced in response to BYMV infection. Ultrastructural investigations of BYMV-infected leaves demonstrated that most chloroplasts with increased stromal area became spherical in shape and some lost their envelopes, either partially or totally. The internal structures of chloroplast, grana and thylakoids were dilated. Two kinds of inclusions were detected in BYMV-infected leaves: straight or slightly curved bands sometimes coiled or looped at the end, and electron opaque crystals with varied shapes. BYMV-infected cells showed lower chloroplast number in comparison to the control. Spraying of SA on faba bean leaves helped to reduce or prevent the harmful effects produced after virus infection. Application of 100 microM SA three days before inoculation restored the metabolism of infected leaves to the levels of healthy controls. SA treatment improved plant health by increasing the photosynthesis rates, pigment contents and levels of other parameters studied similar to control values. Moreover, SA treatment increased plant resistance against BYMV. This was observed through induction of chloroplast number, reduction in percentage of infected plants, decrease in disease severity and virus concentration of plants treated with SA prior to BYMV inoculation. Cells of SA-treated samples showed well-developed chloroplasts with many starch grains and well-organized cell organelles. The present results provide an overview of the negative effects on faba bean leaves due to BYMV infection from physiological and subcellular perspectives. Also, a role of SA involved in induction of resistance against BYMV infection in bean plants is discussed. PMID- 17920159 TI - Identification of enzyme activity that conjugates indole-3-acetic acid to aspartate in immature seeds of pea (Pisum sativum). AB - This study describes the first identification of plant enzyme activity catalyzing the conjugation of indole-3-acetic acid to amino acids. Enzymatic synthesis of indole-3-acetylaspartate (IAA-Asp) by a crude enzyme preparation from immature seeds of pea (Pisum sativum) was observed. The reaction yielded a product with the same Rf as IAA-Asp standard after thin layer chromatography. The identity of IAA-Asp was verified by HPLC analysis. IAA-Asp formation was dependent on ATP and Mg2+, and was linear during a 60 min period. The enzyme preparation obtained after poly(ethylene glycol) 6000 fractionation showed optimum activity at pH 8.0, and the temperature optimum for IAA-Asp synthesis was 30 degrees C. PMID- 17920161 TI - Age-dependent remodelling of retinal circuitry. AB - We have investigated morphological changes in second-order neurons of the mouse retina during aging by using immunohistochemistry and electron microscopy. We observed sprouting of rod bipolar cells dendrites and horizontal cells arborizations: neuronal processes of both neuronal types showed irregular extensions beyond the outer plexiform layer, toward the outer limiting membrane, as well as into the outer nuclear layer (ONL). These processes were first observed in animals of 12 months of age and increased in numbers steadily until 24 months, which represent the last age examined. The ectopic processes are decorated by puncta immunoreactive for pre-synaptic markers typical of photoreceptor terminals juxtaposed to post-synaptic neurotransmitter receptors, demonstrating the presence of the entire molecular machinery of functional synapses. Electron microscopy confirmed that ectopic processes receive synapses from photoreceptor terminals. We conclude that during the second year of life retinal rod bipolar and horizontal cells undergo sprouting and form ectopic synapses in the ONL. PMID- 17920162 TI - [Retinal venous occlusion in Behcet disease]. AB - PURPOSE: The aim of our study is to report three cases of retinal venous occlusion in patients with Behcet disease and to discuss the physiopathology of this vascular accident. METHODS: In this retrospective study, out of a series of 32 patients suffering from Behcet disease, we report three cases of retinal venous occlusion. General and ophthalmologic features are described. Treatment strategy is detailed for each case. RESULTS: There were 2 cases of branch retinal venous occlusion and one case of central retinal venous occlusion. Relapses and retinal neovascularization were the most common complications. CONCLUSION: The retinal venous occlusion in Behcet's disease is a non-common but severe complication. Early and appropriate treatment is required to improve the functional prognosis. PMID- 17920160 TI - The effects of normal aging and ApoE genotype on the levels of CSF biomarkers for Alzheimer's disease. AB - While cerebrospinal fluid (CSF) biomarkers are of use in the prediction and diagnosis of Alzheimer's disease our understanding of the background effects of age and the ApoE genotype is limited. Seventy-eight community-based normal volunteers (mean age 60+/-10 years, range 36-86) were examined to determine the relationships between CSF measures of total tau (T-tau), hyperphosphorylated tau (P-tau 231), amyloid beta (Abeta42/Abeta40 ratio), and isoprostane (IP) with age and ApoE genotype. The results showed that age by epsilon4 genotype interactions were found for P-tau231 (beta=1.82; p<0.05) and IP (beta=1.6; p<0.05). T-tau CSF concentration increased with age. The increasing CSF concentrations of P-tau and IP in epsilon4 carriers suggest that early tauopathy and oxidative stress may be related to the increased risk for AD. The data also suggest that T-tau changes are more age dependent than Abeta changes. The evidence that P-tau231 and IP are the earliest markers for the neuronal damage related to AD awaits longitudinal study. PMID- 17920163 TI - Predicting who will drop out of nursing courses: a machine learning exercise. AB - INTRODUCTION: The concepts of causation and prediction are different, and have different implications for practice. This distinction is applied here to studies of the problem of student attrition (although it is more widely applicable). BACKGROUND: Studies of attrition from nursing courses have tended to concentrate on causation, trying, largely unsuccessfully, to elicit what causes drop out. However, the problem may more fruitfully be cast in terms of predicting who is likely to drop out. METHODS: One powerful method for attempting to make predictions is rule induction. This paper reports the use of the Answer Tree package from SPSS for that purpose. DATA: The main data set consisted of 3978 records on 528 nursing students, split into a training set and a test set. The source was standard university student records. RESULTS: The method obtained 84% sensitivity, 70% specificity, and 94% accuracy on previously unseen cases. DISCUSSION: The method requires large amounts of high quality data. When such data are available, rule induction offers a way to reduce attrition. It would be desirable to compare its results with those of predictions made by tutors using more informal conventional methods. PMID- 17920165 TI - Viral antigen production in cell cultures on microcarriers Bovine parainfluenza 3 virus and MDBK cells. AB - Viral antigens can be obtained from infected mammalian cells cultivated on microcarriers. We have worked out parameters for the production of bovine parainfluenza 3 (PI-3) virus by Mandin-Darby Bovine Kidney (MDBK) cells cultivated on Cytodex 1 microcarriers (MCs) in spinners flasks and bioreactor using fetal bovine serum (FBS) supplemented Eagle minimal essential medium (Eagle MEM). Medium renewal during the cell culture was shown to be crucial for optimal MCs loading (>90% MCs with confluent cell monolayers) and cell growth (2.5 x 10(6)cells/mL and a micro(x) (h(-1)) 0.05). Since cell cultures performed with lower amount of MCs (1g/L), showed good performances in terms of cell loading, we designed batch experiments with a lower concentration of MCs in view of optimizing the cell growth and virus production. Studies of cell growth with lower concentrations of MCs (0.85 g/L) showed that an increase in the initial cell seeding (from 7 to 40 cells/MC) led to a different kinetic of initial cell growth but to comparable final cell concentrations ((8-10)x10(5)cells/mL at 120 h) and cell loading (210-270 cells/MC). Upon infection with PI-3 virus, cultures showed a decrease in cell growth and MC loading directly related to the multiplicity of infection (moi) used for virus infection. Infected cultures showed also a higher consumption of glucose and production of lactate. The PI-3 virus and PI-3 antigen production among the cultures was not significantly different and attained values ranging from, respectively, 7-9 log(10) TCID(50)/mL and 1.5-2.2 OD. The kinetics of PI-3 virus production showed a sharp increase during the first 24h and those of PI-3 antigen increased after 24h. The differential kinetics of PI-3 virus and PI-3 antigen can be explained by the virus sensitivity to temperature. In view of establishing a protocol of virus production and based on the previous experiments, MDBK cell cultures performed under medium perfusion in a bioreactor of 1.2L were infected and the PI-3 virus production in 12L attained 12 log(10) TCID(50). Other than establishing a protocol for PI-3 production in MDBK cell cultures on Cytodex 1, the experiments are proposed as a basis for approaching the development of a virus production protocol in mammalian cells cultivated on microcarriers in bioreactors. PMID- 17920164 TI - Regulation of airway eosinophil and neutrophil infiltration by alpha galactosylceramide in a mouse model for respiratory syncytial virus (RSV) vaccine augmented disease. AB - Respiratory syncytial virus (RSV) is a leading cause of respiratory disease among infants, the elderly and immunocompromised adults. In this study, we assessed the effects of alpha-galactosylceramide, a known immunoregulatory lipid, on liposomal RSV vaccine-induced responses in BALB/c mice subsequently challenged with RSV. Liposomes containing a recombinant fragment of the RSV G protein were prepared with and without alpha-galactosylceramide and used to immunize mice by the intranasal route. The inclusion of alpha-galactosylceramide in the liposomal formulation caused a dramatic reduction in bronchoalveolar lavage neutrophils, but also an increase in eosinophils, following subsequent RSV challenge. The reduction in neutrophils was specific to mice receiving alpha-galactosylceramide containing liposomes and was not reproduced in mice administered liposomes containing another alpha-galactosyl lipid, alpha galactosylphosphatidylglyceroylalkylamine. Lung IL-13 mRNA levels were particularly elevated in mice administered alpha-galactosylceramide-containing liposomes followed by RSV challenge. This study demonstrates a striking ability of alpha-galactosylceramide to modulate the cellular airway infiltrate in mice immunized with liposomal RSV vaccine followed by RSV challenge. PMID- 17920166 TI - A genetically engineered prime-boost vaccination strategy for oculonasal delivery with poly(D,L-lactic-co-glycolic acid) microparticles against infection of turkeys with avian Metapneumovirus. AB - In this study we demonstrated the use of an oculonasally delivered poly(D,L lactic-co-glycolic acid) microparticle (PLGA-MP)-based and genetically engineered vaccination strategy in the avian system. An avian Metapneumovirus (aMPV) fusion (F) protein-encoding plasmid vaccine and the corresponding recombinant protein vaccine were produced and bound to or encapsulated by PLGA-MP, respectively. The PLGA-MP as the controlled release system was shown in vitro to not induce any cytopathic effects and to efficiently deliver the F protein-based aMPV-vaccines to avian cells for further processing. Vaccination of turkeys was carried out by priming with an MP-bound F protein-encoding plasmid vaccine and a booster vaccination with an MP-encapsulated recombinant F protein. Besides the prime boost F-specific vaccinated birds, negative control birds inoculated with a mock MP prime-boost regimen as well as non-vaccinated birds and live vaccinated positive control birds were included in the study. The MP-based immunization of turkeys via the oculonasal route induced systemic humoral immune reactions as well as local and systemic cellular immune reactions, and had no adverse effects on the upper respiratory tract. The F protein-specific prime-boost strategy induced partial protection. After challenge the F protein-specific MP-vaccinated birds showed less clinical signs and histopathological lesions than control birds of mock MP-vaccinated and non-vaccinated groups did. The vaccination improved viral clearance and induced accumulation of local and systemic CD4+ T cells when compared to the mock MP-vaccination. It also induced systemic aMPV-neutralizing antibodies. The comparison of mock- and F protein-specific MP-vaccinated birds to non-vaccinated control birds suggests that aMPV-specific effects as well as adjuvant effects mediated by MP may have contributed to the overall protective effect. PMID- 17920168 TI - Vaccine-preventable influenza disease burden from clinical trials of Vaxigrip - an inactivated split virion influenza vaccine - supports wider vaccine use. AB - Trivalent inactivated split influenza virus vaccine has been used for more than 35 years, and is currently licensed in over 100 countries. To determine vaccine preventable influenza burden in different populations and geographic regions, we reviewed studies of vaccine effectiveness against non-specific outcomes such as upper respiratory infection, hospitalization, and death in addition to confirmed microbiologically confirmed influenza. The vaccine-preventable disease incidence was high in most studies, regardless of the outcome or population evaluated. This indicates that routine influenza vaccination can improve overall population health under a broad range of circumstances. PMID- 17920167 TI - Immunisation of sheep against heartwater in The Gambia using inactivated and attenuated Ehrlichia ruminantium vaccines. AB - Heartwater (cowdriosis) is a disease of ruminants caused by a rickettsial pathogen Ehrlichia ruminantium and transmitted by ticks of the genus Amblyomma. The purpose of this work was to evaluate the protective efficacies of inactivated and attenuated vaccines to protect sheep against heartwater in The Gambia. An inactivated vaccine, prepared from E. ruminantium (Gardel stock), and a live attenuated vaccine from E. ruminantium (Senegal stock), were evaluated in two independent on-station trials. A local stock of E. ruminantium (Kerr Seringe) was used as challenge material. Inactivated and live attenuated vaccines provided 43% and 100% protection, respectively, against virulent needle challenge. In a subsequent field trial, the attenuated vaccine protected 75% of sheep against virulent tick challenge, which was fatal for all control sheep. Quantification by real-time PCR showed that an immunising dose of approximately 23,000 attenuated E. ruminantium organisms was sufficient. Moreover, restriction fragment length polymorphism (RFLP) analysis indicated that the local Kerr Seringe genotype caused mortality amongst control sheep, whereas fatalities in the vaccinated group could be attributed to a different genotype. PMID- 17920169 TI - Immunization with radioattenuated yeast cells of Paracoccidioides brasiliensis induces a long lasting protection in BALB/c mice. AB - Paracoccidioides brasiliensis is the fungus agent of paracoccidioidomycosis, a chronic systemic disease prevalent in Latin America. The aim of the present work was to evaluate the protection elicited by the immunization of BALB/c mice with radioattenuated yeast cells of P. brasiliensis. The immunization promoted a long lasting protection against highly infective yeast forms of P. brasiliensis. A 99.5% decrease in CFUs recovery was verified 90 days post challenge. At the same time the levels of IgG2a and IFN-gamma were high while a very low production of IL-10 and IL-5 was verified, suggesting that a Th1 pattern was dominant. This work shows the potential of radioattenuated yeast cells for the development of vaccines against fungi infections. PMID- 17920170 TI - More vaccines for children? Parents' views. AB - AIM: To investigate parents' views regarding potential new vaccines. We examined attitudes towards severity of specific infections, acceptability of potential vaccines and preferences for the number of injections they would want their child to receive on any one occasion. DESIGN: Cross sectional survey. METHODS: Parents of children aged 18-24 months in three Primary Care Trusts in England were asked to complete a questionnaire. RESULTS: Of the 859 parents who responded (38%), over 90% believed that the vaccines currently on offer prevent disease always or almost always. Of those who rated meningitis as serious or very serious, 84% would accept a vaccine against meningococcal group B infection and 69% against pneumococcal infection. Only 34% of those who said their child would make a full recovery from chicken pox would accept a vaccine. Over half the respondents preferred new vaccines to be given separately. Fifty seven percent of parents would not want their child to have more than two injections per clinic visit. CONCLUSIONS: Parents' views on the severity of illness influence their acceptance of a new vaccine. Their preference for as few injections as possible at a single clinic visit needs to be reconciled with their concerns over the use of combination vaccines. PMID- 17920171 TI - Perceptions of motherhood: The effect of experience and knowledge on midwifery students. AB - OBJECTIVE: to explore the factors that influence student midwives' constructs of childbearing, before and during their undergraduate midwifery programme. DESIGN: a naturalistic, qualitative study. SETTING: a university in the East Midlands, UK. PARTICIPANTS: 58 women registered on a 3-year midwifery education programme. MEASUREMENTS: focus groups were conducted at programme commencement and at 9-12 monthly intervals with two cohorts of midwifery students who were separated into groups of mothers and non-mothers (32 focus groups in total). This paper draws on data from the nine focus groups held at the start of the students' programme. FINDINGS: the main themes that emerged from the data were in relation to image during pregnancy, expectations/experiences of childbirth and parenting. In particular, students believed that pregnancy and childbirth should be special. They suggested that a lack of knowledge about sexuality and choice options affected women's ability to be in control. Although normality was the students' expectation of childbirth, they also assumed that hospital birth was the norm. They were unsure whether the baby's father was the best birth partner. Their mothers were suggested as likely to be more supportive, but there was lack of agreement regarding whether they were the best parenting role models. Students also said that there was a lack of positive images of breast feeding. The overall motivation to become midwives was 'to make a difference'. CONCLUSIONS: students need to be facilitated early in their programme to explore their belief systems and constructs of childbearing critically so that they are equipped to support parents to have a positive experience, whether childbirth is normal or complex, and so that they can cope with any dissonance between their own expectations and the uncertainties and realities of practice. PMID- 17920172 TI - Perception of risk in relation to ultrasound screening for Down's syndrome during pregnancy. AB - OBJECTIVE: to explore how information about being at risk of carrying a fetus with Down's syndrome was understood, and whether the actual risk and the woman's perception of risk was associated with worry or depressive symptoms during and after pregnancy. DESIGN AND SETTING: observational study. The sample was drawn from the intervention group of a Swedish randomised controlled trial of ultrasound screening for Down's syndrome by nuchal translucency measurement. MEASUREMENTS: data were collected by three questionnaires. Questions were asked about recall of the risk score and perception of risk. The Cambridge Worry Scale and the Edinburgh Postnatal Depression Scale measured worry and depressive symptoms, respectively, on all three occasions. FINDINGS: of the 796 women who provided data for this study, one in five was unaware that the risk score was noted in her case record. In total, 620 women stated that they had received a risk score, but only 64% of them recalled the figure exactly or approximately. The actual risk was associated with the perceived risk, but of the 31 women who perceived the risk to be high, only 14 were actually at high risk. A high-risk score was not associated with worry or depressive symptoms in mid-pregnancy, in contrast to a woman's own perception of being at high risk. Two months postpartum, no associations were found between maternal emotional well-being and actual or perceived risk. CONCLUSIONS: information about fetal risk is complicated and women's perception of risk does not always reflect the actual risk, at least not when presented as a numerical risk score. The possibility that the information may cause unnecessary emotional problems cannot be excluded. IMPLICATIONS FOR PRACTICE: caregivers should ascertain that information about fetal risk is interpreted correctly by pregnant women. PMID- 17920173 TI - Flexor tendon repair using the two-strand side-locking loop technique to tolerate aggressive active mobilization immediately after surgery. AB - BACKGROUND: Early mobilization after tendon repair decreases adhesion formation and improves repair-site strength. We investigated whether the two-strand side locking loop technique would tolerate aggressive active mobilization immediately after surgery. METHODS: Twelve flexor digitorum profundus tendons of the porcine forelimbs were sutured by the two-strand side-locking loop technique with a cross stitch epitendinous repair (Group A), and by the 8-strand repair method with a simple running suture (Group B). Gaps and residual tensile strength after cyclic loadings of 3-50 N (for 10,000 rounds) were measured. FINDINGS: Gaps after cyclic loading in Group A were 0.5+/-0.3 and 1.2+/-0.8 mm while those in Group B were 3.5+/-0.8 and 5.2+/-1.2 mm at 3 and 50 N, respectively. In addition, the respective residual tensile strength of Groups A and B were 207.1+/-15.2 and 84.2+/-18.3N. INTERPRETATION: A combination of the two-strand side-locking loop technique with cross-stitch epitendinous repair served as the optimum suture method in establishing safe and early active mobilization without the aid of a specialized rehabilitation staff. PMID- 17920174 TI - Variation in PAH inputs and microbial community in surface sediments of Hamilton Harbour: implications to remediation and monitoring. AB - Variations in concentrations of polycyclic aromatic hydrocarbons (PAHs) and microbial community indicators were investigated in representative highly contaminated and less contaminated surface sediment sites of Hamilton Harbour. Inputs of PAH to the upper 3cm of sediments up to four times the average upper sediment concentrations were observed. Associated PAH fingerprint profiles indicated that the source was consistent with the PAH source to the industrial region of the harbour. Increased PAH loadings were associated with decreased bacterial populations as indicated by phospholipid fatty acid (PLFA) concentrations. However, relatively minor impacts on overall community composition were indicated. Porewater methane concentrations and isotopic data indicated a difference in the occurrence of methane oxidation between the two sites. This study confirms temporally limited transport of contaminants from highly impacted regions as a vector for contaminants within the harbour and the impact on microbial carbon cycling and bed stability. PMID- 17920175 TI - Reduced nitrogen in ecology and the environment. AB - Since the beginning of the 19th century humans have increasingly fixed atmospheric nitrogen as ammonia to be used as fertilizer. The fertilizers are necessary to create amino acids and carbohydrates in plants to feed animals and humans. The efficiency with which the fertilizers eventually reach humans is very small: 5-15%, with much of the remainder lost to the environment. The global industrial production of ammonia amounts to 117 Mton NH(3)-Nyear(-1) (for 2004). By comparison, we calculate that anthropogenic emissions of NH(3) to the atmosphere over the lifecycle of industrial NH(3) in agriculture are 45.3 Mton NH(3)-Nyear(-1), about half the industrial production. Once emitted ammonia has a central role in many environmental issues. We expect an increase in fertilizer use through increasing demands for food and biofuels as population increases. Therefore, management of ammonia or abatement is necessary. PMID- 17920176 TI - Effects of racial density and income incongruity on pregnancy outcomes in less segregated communities. AB - A previous publication in this journal documented a decreased risk of adverse birth outcomes when African-American women have a positive income incongruity (defined as mothers living in a census tract with a higher household income than would be expected based on their individual education and marital status) and live in a census tract with "predominantly African-American" residents [Pickett, K. E., Collins, J. W. Jr., Masi, C. M., & Wilkinson, R. G. (2005). The effects of racial density and income incongruity on pregnancy outcomes. Social Science & Medicine, 60(10), 2229-2238.]. The communities included in that study were from Chicago and were highly segregated by race. Our objective was to repeat this analysis in a less severely segregated environment: two urban counties (Wake and Durham) in central North Carolina. Rather than assuming an absence of knowledge about the effects of interest, we used the previously published results to inform our prior distributions in a Bayesian logistic regression analysis. This approach, which is analogous to a meta-analysis of the two studies, revealed a protective effect of positive income incongruity for African-American women living in census tracts with high relative African-American density across a much wider range of residential segregation patterns. Positive income incongruity was not associated with a decreased risk of low birth weight or preterm delivery for women living in tracts with a low relative density of African-Americans. These estimates are comparable to those that might have been observed had the original authors included a much more diverse set of communities with respect to degree of segregation, and so these new results provide important information about the generality of this intriguing finding. PMID- 17920178 TI - Women's management of menstrual symptoms: findings from a postal survey and qualitative interviews. AB - This paper draws on postal questionnaire and qualitative interview data to investigate women's experiences of and responses to menstrual symptoms. Respondents were drawn from general practices in the Lothian region of Scotland. They described a range of strategies to manage menstrual symptoms, learnt both from personal experience and discussions with others. While most women had spoken to others about periods, this more often related to matters such as changes with age, or pain management, whereas heaviness of blood loss was not usually a subject for discussion. When asked about formal help-seeking women described it as a step reluctantly taken, only if attempts to self manage failed, and that often it was opportunistic, occurring when consulting for something else. It was apparent that women thought of menstrual symptoms, even when felt to be problematic, as 'part and parcel' of female life and not a legitimate reason for adopting illness behaviour. The way that women spoke of managing symptoms suggested that responding to menstrual symptoms takes place within a moral framework and that the only 'legitimate' or 'virtuous' response was a stoical one. Our findings suggest that women with debilitating menstrual symptoms may not have access to the best information, advice or treatments for this. A supported self care approach could help, combining access to high quality information about what is usual and unusual for women, with information about what can help, what treatments are available from which sources and guidance on when and how to seek professional advice. If available in a range of formats, such as features in women's magazines, health-related web-sites and information leaflets, such an approach may help women self-manage their debilitating symptoms even better. PMID- 17920177 TI - Education, income and ethnic differences in cumulative biological risk profiles in a national sample of US adults: NHANES III (1988-1994). AB - Data from the nationally representative US National Health and Nutrition Examination Survey (NHANES) III cohort were used to examine the hypothesis that socio-economic status is consistently and negatively associated with levels of biological risk, as measured by nine biological parameters known to predict health risks (diastolic and systolic blood pressure, pulse, HDL and total cholesterol, glycosylated hemoglobin, c-reactive protein, albumin and waist-hip ratio), resulting in greater cumulative burdens of biological risk among those of lower education and/or income. As hypothesized, consistent education and income gradients were seen for biological parameters reflecting cardiovascular, metabolic and inflammatory risk: those with lower education and income exhibiting greater prevalence of high-risk values for each of nine individual biological risk factors. Significant education and income gradients were also seen for summary indices reflecting cumulative burdens of cardiovascular, metabolic and inflammatory risks as well as overall total biological risks. Multivariable cumulative logistic regression models revealed that the education and income effects were each independently and negatively associated with cumulative biological risks, and that these effects remained significant independent of age, gender, ethnicity and lifestyle factors such as smoking and physical activity. There were no significant ethnic differences in the patterns of association between socio-economic status and biological risks, but older age was associated with significantly weaker education and income gradients. PMID- 17920179 TI - Evidence for the involvement of the monoaminergic system in the antidepressant like action of two 4-amine derivatives of 10,11-dihydro-5H-dibenzo [a,d] cycloheptane in mice evaluated in the tail suspension test. AB - Our previous study described the synthesis of 4-amine derivatives of 10,11 dihydro-5H-dibenzo-alkylamine-cycloheptane, 4-amine (3-N,N-dimethylpropylamine) 10,11-dihydro-5H-dibenzo[a,d] cycloheptane-5-one (ADDCH1), and 1,2,3,4,8,9 hexahydro-dibenzocycloheptane[4,4a,5-ef]1,4-diazepin (ADDCH2), and the characterization of their antidepressant-like effect in the forced swimming test in mice. This study investigated the involvement of monoaminergic pathways in the antidepressant-like effect of these compounds in mice evaluated in the tail suspension test (TST), another animal model to screen antidepressant drugs. Our results show that the immobility time in the TST was significantly reduced by ADDCH1 (15 to 50 mg/kg, i.p.) or ADDCH2 (30 and 50 mg/kg, i.p.). The antidepressant-like effect of ADDCH1 (30 mg/kg, i.p.) in the TST was prevented by pre-treatment of mice with methysergide (2 mg/kg, i.p.), a non-selective serotonin receptor antagonist, p-chlorophenylalanine methylester (pCPA, 100 mg/kg, i.p.), an inhibitor of serotonin synthesis, prazosin (62.5 microg/kg, i.p.), an alpha1-adrenoceptor antagonist, or yohimbine (1 mg/kg, i.p.), an alpha2 adrenoceptor antagonist. In contrast, the antidepressant-like effect of ADDCH2 was antagonized only by yohimbine (1 mg/kg) or haloperidol (50 microg/kg, i.p.), a dopamine D2/D3/D4 receptor antagonist, and was not affected by methysergide, pCPA or prazosin. Altogether, the present results strongly suggest the differential involvement of monoaminergic systems, serotonin/noradrenaline (ADDCH1) and noradrenaline/dopamine (ADDCH2) pathways, respectively, in the antidepressant-like effect of dibenzosuberone compounds. PMID- 17920180 TI - An interaction between the norepinephrine transporter and monoamine oxidase A polymorphisms, and novelty-seeking personality traits in Korean females. AB - The personality traits associated with the noradrenergic system have not yet been clearly established. In the present study, we investigated the variable number of tandem repeats (VNTR) polymorphism of the norepinephrine transporter (NET) and monoamine oxidase A (MAOA), which are major components of the adrenergic system, to elucidate their relationship with personality. A total of 245 normal female Koreans (age 23.05+/-3.07 years, mean+/-SD) volunteered to take part in this study. They filled out a Korean version of the Temperament and Character Inventory (TCI) and were genotyped for the NET and MAOA-VNTR; the NET T-182C and MAOA-uVNTR polymorphisms were checked. We found significant main effect of NET genotype on novelty seeking (NS) score (F=5.43, p=0.021) and significant interaction between the NET and MAOA-uVNTR polymorphisms on NS score (F=11.06, p=0.001). However, there were no relationship between MAOA-uVNTR polymorphisms and NS score, and no association with other temperamental dimensions and these two polymorphisms. Our findings suggest that this functional polymorphism in the noradrenergic gene is associated with novelty seeking in Korean females. PMID- 17920181 TI - [Letters to the editor]. PMID- 17920182 TI - The effects of methylmercury on motor activity are sex- and age-dependent, and modulated by genetic deletion of adenosine receptors and caffeine administration. AB - Adenosine and its receptors are, as part of the brain stress response, potential targets for neuroprotective drugs. We have investigated if the adenosine receptor system affects the developmental neurotoxicity caused by the fish pollutant methylmercury (MeHg). Behavioral outcomes of low dose perinatal MeHg exposure were studied in mice where the A(1) and A(2A) adenosine receptors were either partially blocked by caffeine treatment or eliminated by genetic modification (A(1)R and A(2A)R knock-out mice). From gestational day 7 to day 7 of lactation dams were administered doses that mimic human intake via normal diet, i.e. 1microM MeHg and/or 0.3g/l caffeine in the drinking water. This exposure to MeHg resulted in a doubling of brain Hg levels in wild type females and males at postnatal day 21 (PND21). Open field analysis was performed at PND21 and 2 months of age. MeHg caused time-dependent behavioral alterations preferentially in male mice. A decreased response to amphetamine in 2-month-old males pointed to disturbances in dopaminergic functions. Maternal caffeine intake induced long lasting changes in the offspring evidenced by an increased motor activity and a modified response to psychostimulants in adult age, irrespectively of sex. Similar alterations were observed in A(1)R knock-out mice, suggesting that adenosine A(1) receptors are involved in the alterations triggered by caffeine exposure during development. Perinatal caffeine treatment and, to some extent, genetic elimination of adenosine A(1) receptors, attenuated the behavioral consequences of MeHg in males. Importantly, also deletion of the A(2A) adenosine receptor reduced the vulnerability to MeHg, consistent with the neuroprotective effects of adenosine A(2A) receptor inactivation observed in hypoxia and Parkinson's disease. Thus, the consequences of MeHg toxicity during gestation and lactation can be reduced by adenosine A(1) and A(2A) receptor inactivation, either via their genetic deletion or by treatment with their antagonist caffeine. PMID- 17920183 TI - Pharmacologic treatment of male stress urinary incontinence: systematic review of the literature and levels of evidence. AB - OBJECTIVE: Stress urinary incontinence (SUI) occurs in men and women. Pharmacologic treatment of female SUI has been beneficiary but the role of drug treatment in male SUI is controversial. This review evaluates the drug classes, the effects of these drugs in trials with male SUI, and the levels of evidence of the individual trials. METHODS: A systematic literature review was conducted searching for studies on male SUI published between 1966 and 2007. In retrieved articles, reference lists were hand-searched to identify additional articles. The level of evidence was judged according to the Oxford classification. RESULTS: This search found nine articles providing evidence that alpha-adrenoceptor agonists (ephedrine, phenylpropanolamine, midodrine), beta2-adrenoceptor agonists (clenbuterol), and serotonin-noradrenaline reuptake inhibitors (imipramine, duloxetine) have a potentially beneficial effect on male SUI. Most trials used only small numbers of patients and a mixed study population (men and women). The evidence level of most studies was low due to the lack of randomisation (case series or cohort studies, level 4). In contrast, the first high-level study to provide evidence in the treatment of male SUI was with duloxetine, a selective serotonin and noradrenaline reuptake inhibitor (randomised controlled study, level 1b). However, only one well-designed study has been published so far. CONCLUSIONS: One high-level study with duloxetine in combination with pelvic floor muscle training shows preliminary but promising results in the treatment of male SUI. These results have to be confirmed in larger and well-designed trials to allow definite recommendations for the pharmacologic treatment of male SUI. PMID- 17920184 TI - EAU guidelines on prostate cancer. AB - OBJECTIVES: To present a summary of the 2007 version of the European Association of Urology (EAU) guidelines on prostate cancer (PCa). METHODS: A literature review of the new data emerging from 2004 to 2007 was performed by the working panel. The guidelines have been updated, and the level of evidence/grade of recommendation was added to the text based on a systematic review of the literature, which included a search of online databases and bibliographic reviews. RESULTS: A full version is available at the EAU Office or at www.uroweb.org. Systemic prostate biopsy under ultrasound guidance is the preferred diagnostic method. Active treatment is mostly recommended for patients with localized disease and a long life expectancy, with radical prostatectomy being shown to be superior to watchful waiting in a prospective randomized trial. Nerve-sparing radical prostatectomy represents the approach of choice in organ confined disease; neoadjuvant androgen deprivation demonstrates no improvement of outcome variables. Radiation therapy should be performed with at least 72 and 78 Gy in low-risk and intermediate- to high-risk PCa, respectively. Monotherapeutic androgen deprivation is the standard of care in metastatic PCa; intermittent androgen deprivation might be an alternative treatment option for selected patients. Follow-up is largely based on prostate-specific antigen and a disease specific history with imaging only indicated when symptoms occur. Cytotoxic therapy with docetaxel has emerged as the reference treatment for metastatic hormone-refractory PCa. CONCLUSIONS: The knowledge in the field of PCa is rapidly changing. These EAU guidelines on PCa summarize the most recent findings and put them into clinical practice. PMID- 17920185 TI - Re: Paula M.J. Moonen, Judith M.J.E. Bakkers, Lambertus A.L.M. Kiemenay et al. Human papilloma virus DNA and p53 mutation analysis on bladder washes in relation to clinical outcome of bladder cancer. Eur Urol 2007;52:464-9. PMID- 17920186 TI - ERRbeta: a potent inhibitor of Nrf2 transcriptional activity. AB - The orphan nuclear receptor, estrogen-related receptor beta (ERRbeta), shares a high degree of amino acid identity with estrogen receptor alpha (ERalpha). Although ERRbeta has been shown to be critical in embryo development, little is known about its functions and target genes. Here we report that the newly identified and most common human ortholog of ERRbeta--short-form hERRbeta (SFhERRbeta) potently represses the transcriptional activity of NF-E2 Related Factor 2 (Nrf2) on antioxidant response element (ARE)-mediated gene expression. Nrf2 is a main regulator of the expression of phase II detoxifying enzymes and antioxidant proteins in the cellular protection against oxidative stress. SFhERRbeta is the most potent inhibitor of Nrf2 transcriptional activity among the three ERR family members, ERRalpha, ERRbeta and ERRgamma. Additional analyses revealed that SFhERRbeta repressed Nrf2 activity likely through physical interaction in a complex with Nrf2, not by competing for the ARE DNA-binding sites, nor by decreasing Nrf2 protein concentration. By confocal immunofluorescence microscopy, SFhERRbeta alters the subcellular localization of Nrf2. Analyses using SFhERRbeta deletion mutants showed that SFhERRbeta interacts with Nrf2 through multiple sites. Our findings suggest that ERRbeta plays a novel functional role in the Nrf2-ARE pathway. By acting as a repressor of Nrf2, ERRbeta may be useful as a therapeutic target in cancer chemoprevention studies. PMID- 17920187 TI - White matter hyperintense lesions in genetically proven spinocerebellar ataxia 8. AB - We report two brothers with a progressive cerebellar syndrome due to spinocerebellar ataxia type 8 (SCA8). In addition to severe cerebellar atrophy, both had prominent white matter hyperintensities on cranial MRI. This is the first report of white matter hyperintensities on cranial MRI in patients with SCA8. A disorder due to a similar molecular basis, myotonic dystrophy 1 (DM1), is known to have white matter hyperintensities on cranial MRI. Cognitive impairment is well described in DM1 and is being recognized in SCA8. The significance of these associations is discussed. PMID- 17920188 TI - Transient global amnesia after breathing hyperoxic mixtures in otherwise regular dives. AB - The diagnosis of decompression sickness may be difficult, particularly when patients present with atypical clinical manifestations after dives going on uneventfully and with full adherence to safety parameters. The case reports involve two divers who presented with transient global amnesia (TGA) immediately after surfacing from otherwise regular dives during which they had been breathing hyperoxic mixtures (Nitrox). A wide spectrum of symptoms of neural dysfunction following hyperoxic mixtures have been reported, including cases of memory impairment related to previous generalized convulsions. After a review of the literature, however, we found no previous reports of TGA, which is very unlikely to result from an epileptic mechanism, as an adverse effect of breathing hyperoxic mixtures. PMID- 17920189 TI - Amyotrophic lateral sclerosis patients and ocular ptosis. AB - Ptosis is not a feature observed in amyotrophic lateral sclerosis (ALS). We describe two old women with bulbar-onset ALS and rapid progression, in whom ptosis and diplopia were noted. They did not improve on pyridostigmine or steroids. Antibodies against acetylcholine receptor were negative, thymoma was excluded, but neurophysiological showed marked neuromuscular transmission failure in orbicularis oculi. We discuss the association between ALS and ocular myasthenia gravis in these cases. PMID- 17920190 TI - Magnetic resonance imaging of idiopathic carpal tunnel syndrome: correlation with clinical findings and electrophysiological investigation. AB - OBJECTIVE: To compare clinical evaluation, electrophysiological investigation and magnetic resonance findings in assessing the severity of idiopathic carpal tunnel syndrome. PATIENTS AND METHODS: Seventy-four patients with idiopathic carpal tunnel syndrome were prospectively recruited. Clinical evaluation included symptoms severity score and two-point discrimination, sensory and motor nerve conduction velocities were determined by electroneuromyography and imaging parameters were obtained after wrist magnetic resonance. The Wilcoxon test was used to define the differences between measurements of median nerve area. The Pearson and Spearman correlation tests were used to determine the relationships between all the measured parameters. RESULTS: Cross-sectional area of median nerve was smaller at hamate level than at radio-ulnar joint and pisiform levels (p<0.001). With exception of median nerve area at hamate level, there was a lower degree of correlation between MRI parameters and findings obtained by clinical assessments and electrophysiological measurements. The median nerve area at hamate level correlated negatively with duration of symptoms, two-point discrimination, symptoms severity score and positively with sensory nerve conduction velocity (p<0.01). CONCLUSION: In patients with idiopathic carpal tunnel syndrome, median nerve area measured by wrist magnetic resonance at hamate level may be considered as a valuable indicator to grading the severity of disease. PMID- 17920191 TI - Neuroimage: disseminated nocardiosis. PMID- 17920192 TI - Genetic analysis of head and neck squamous cell carcinoma using comparative genomic hybridisation identifies specific aberrations associated with laryngeal origin. AB - Head and neck squamous cell carcinoma (HNSCC) demonstrates significant differences in the biological and clinical behaviour of tumours found at different sub-sites. We investigated the genetic profiles of 68 carcinomas (larynx n=35, hypopharynx n=19, oropharynx n=14) using chromosomal comparative genomic hybridisation in order to identify sub-site specific differences. Multiple genetic aberrations were found throughout the tumour genomes, including +3q (82%), -3p (75%), +8q (66%), +5p (49%), +7q (49%), +1q (47%), -4p (46%), -11q (46%), -13q (46%), -5q (44%), +11q (43%) and +12p (43%). The mean number of chromosomal arms with at least one aberration was 15. Laryngeal carcinomas (LSCC) were found to have significantly more aberrations on chromosomal arms than oropharyngeal carcinomas (OpSCC); (mean of 17 vs. 11, respectively (p=0.011). It was noted that -4p, +8q, +12q, and -18q were significantly associated with LSCC when compared with both hypopharyngeal SCC (HpSCC) and OpSCC. HpSCC was significantly associated with -2q whereas no aberrations were found to be significantly associated with OpSCC. In conclusion a large number of common chromosomal aberrations are associated with HNSCC however in addition further aberrations are significantly associated with individual sub-sites of head and neck cancer. These aberrations may be responsible for the diverse biological behaviour of these different tumour types. Further research is required to identify the specific genes associated with these chromosomal regions and evaluate their individual impact on disease progression. PMID- 17920193 TI - Hypoxia-inducible factors: crosstalk between their protein stability and protein degradation. AB - As a transcription factor family dependent of oxygen, hypoxia-inducible factors (HIFs) play important roles in organic and cellular homeostasis, which are master regulators of tumorigenesis, angiogenesis and embryogenesis. Up to date, some known posttranslational modifications, such as hydroxylation, acetylation, phosphorylation and S-nitrosylation, have been proved to influence protein stability and transcriptional activity of HIFs. On the other hand, HIFs can be physiologically degraded through ubiquitin-dependent or -independent proteasome pathway. Two biochemical processes involve in many modulators, which are correlated and compose an intricate mechanism to control the biological function of HIFs systemically. Herein, we review and discuss the diverse roles of involved modulators in both HIFs stability and degradation, describe the potential link between the two molecular scenarios. PMID- 17920195 TI - D-serine regulates CREB phosphorylation induced by NMDA receptor activation in Muller glia from the retina. AB - D-serine is an N-methyl-D-aspartate (NMDA) receptor coagonist predominantly produced by glial cells in the brain and the retina. Whereas a role for D-serine as a modulator of NMDA receptors in neurons has been suggested, its function in glial cells has not been analyzed. We here show that D-serine modulates gene expression in Muller glial cells from the retina through the induction of transcription factor CREB phosphorylation and the expression of the immediate early gene c-fos. Pharmacological analysis indicates that D-serine effect involves NMDA receptor activation. Comparison of the effect of D-serine in Muller cells, hippocampal astrocytes and hippocampal neurons suggests that D-serine could function as a retinal NMDA receptor coagonist activating functionally relevant transcription factor pathways in glial cells. PMID- 17920194 TI - Role of glutamatergic receptors located in the nucleus raphe magnus on antinociceptive effect of morphine microinjected into the nucleus cuneiformis of rat. AB - Neurons in the nucleus cuneiformis (CnF), located just ventrolateral to the periaqueductal gray, project to medullary nucleus raphe magnus (NRM), which is a key medullary relay for descending pain modulation and is critically involved in opioid-induced analgesia. Previous studies have shown that antinociceptive response of CnF-microinjected morphine can be modulated by the specific subtypes of glutamatergic receptors within the CnF. In this study, we evaluated the role of NMDA and kainate/AMPA receptors that are widely distributed within the NRM on morphine-induced antinociception elicited from the CnF. Hundred and five male Wistar rats weighing 250-300 g were used. Morphine (10, 20 and 40 microg) and NMDA receptor antagonist, MK-801 (10 microg) or kainate/AMPA receptor antagonist, DNQX (0.5 microg) in 0.5 microl saline were stereotaxically microinjected into the CnF and NRM, respectively. The latency of tail-flick response was measured at set intervals (2, 7, 12, 17, 22, 27 min after microinjection) by using an automated tail-flick analgesiometer. The results showed that morphine microinjection into the CnF dose-dependently causes increase in tail-flick latency (TFL). MK-801 microinjected into the NRM, just 1 min before morphine injection into the CnF, significantly attenuated antinociceptive effects of morphine. On the other hand, DNQX microinjected into the NRM, significantly increased TFL after local application of morphine into the CnF. We suggest that morphine related antinociceptive effect elicited from the CnF is mediated, in part, by NMDA receptor at the level of the NRM whereas kainite/AMPA receptor has a net inhibitory influence at the same pathway. PMID- 17920196 TI - The treatment of neuropathic pain: from hubris to humility. PMID- 17920197 TI - Effects of specimen tilt in ADF-STEM imaging of a-Si/c-Si interfaces. AB - Annular dark field scanning transmission electron microscopy (ADF-STEM) imaging of a crystal depends strongly on specimen orientation, but for an amorphous sample it is insensitive to orientation changes. To fully investigate the effects of specimen tilt, an interface of amorphous Si (a-Si) and crystalline Si (c-Si) was rotated systematically off a zone axis in a STEM equipped with low-angle ADF (LAADF) and high-angle ADF (HAADF) detectors. The change of relative intensity across the interface shows very different trends in the LAADF and the HAADF images upon tilting. More importantly, it is found that the HAADF signal decreases much more rapidly when tilted off a zone axis than does the LAADF signal. The high-resolution lattice fringes also disappear much faster in the HAADF image than in the LAADF image. These trends reflect the fact that the channeling peaks that are responsible for scattering into the HAADF detector decrease more quickly upon tilting than the lower angle scattering to the LAADF detector does. PMID- 17920198 TI - Moisture requirements for the free-living development of Haemonchus contortus: quantitative and temporal effects under conditions of low evaporation. AB - The key influence of moisture availability on development of the free-living stages of Haemonchus contortus is well-documented, although quantitative relationships between moisture variables and development are poorly defined. A factorial experiment (3x2x2) was conducted in programmable incubators to determine the effects of amount and distribution of simulated rainfall on H. contortus development at low evaporation rates (approximately 2 mm/day) under temperatures typical of summers on the Northern Tablelands of NSW, Australia. Sheep faeces containing H. contortus eggs were placed in experimental units containing sterile soil and had one of the three amounts (12, 24 and 32 mm) of simulated rainfall applied, in either a single event on the day after deposition (d 1) or three split events over 6 days (d 1, 3 and 6). Treatments were applied either in week 1 only, or in weeks 1 and 2. Recovery of infective larvae (L3) at 4, 7 and 14 days post-contamination increased with each incremental amount of simulated rainfall over the range of 12-32 mm and was significantly higher under the single "rain" event (2.8%), compared with the three smaller, split events (1.9%). The second application of simulated rainfall in week 2 had only a very small influence on L3 recovery, suggesting that the majority of development to L3 occurred in response to simulated rainfall events in the first 7 days. Both faecal moisture content and the cumulative ratio of precipitation and evaporation (P/E) were strongly and positively correlated with recovery of L3. Recovery of L3 from treatments, which received simulated rainfall in week 1 only was best described by P/E at d 5. Whether the relationships observed in this study hold under field conditions, where macroclimatic conditions such as evaporation rate are substantially more limiting to free-living development, is yet to be determined. PMID- 17920199 TI - Plant collagenase: unique collagenolytic activity of cysteine proteases from ginger. AB - Two cysteine proteases, GP2 and GP3, have been isolated from ginger rhizomes (Zingiber officinale). GP2 is virtually identical to a previously identified ginger protease GPII [K.H. Choi, and R.A. Laursen, Amino-acid sequence and glycan structures of cysteine proteases with proline specificity from ginger rhizome Zingiber officinale, Eur. J. Biochem. 267 (2000) 1516-1526.], and cleaves native type I collagen at multiple discrete sites, which are in the interior of the triple helical region of this molecule. In reaction with proline-containing peptides GP2 shows preference for Pro in the P2 position, and at least 10-fold higher efficiency of hydrolysis than papain. Comparison of models of GP2 and GP3 with the crystal structure of papain shows that the three enzymes have different S2 pocket structures. The S2 pocket in GP2 and GP3 is half the size of that of papain. GP2 is the only reported plant cysteine protease with a demonstrated ability to hydrolyse native collagen. The results support a role for ginger proteases as an alternative to papain, in commercial applications such as meat tenderization, where collagen is the target substrate. PMID- 17920200 TI - Trypanosoma cruzi as a model system to study the expression of exogenous genes coding for polyamine biosynthetic enzymes. Induction of DFMO resistance in transgenic parasites. AB - Trypanosoma cruzi, the etiologic agent of Chagas' disease, is a polyamine auxotroph organism because its genome contains neither ornithine decarboxylase (ODC) nor arginine decarboxylase (ADC) genes, presumably lost during evolution. After transformation with a recombinant plasmid bearing the complete coding region of Crithidia fasciculata ODC gene, the transgenic parasites were able to synthesize putrescine and simultaneously became susceptible to alpha difluoromethylornithine (DFMO), an irreversible inhibitor of ODC. We have studied the emergence of DFMO-resistant T. cruzi after one-step selection of ODC transformed parasites cultivated in the presence of high levels of the drug (5 mM). Our results have indicated a duplication of the ODC gene copy number in the drug-resistant cell line. The ODC transcripts and the corresponding translation products showed very significant increases (about 7- and 25-fold, respectively) in DFMO-resistant parasites, while the ODC enzymatic activity was 5 times higher than in drug-sensitive T. cruzi. The unequal increases of ODC protein and enzymatic activity in DFMO-resistant protozoa strongly suggest that in addition to gene amplification and enhanced transcription and translation, the assembly of ODC polypeptide chains into dimeric active enzyme molecules might also contribute to regulate the development of DFMO resistance. PMID- 17920201 TI - A novel ribonuclease with antiproliferative activity from fresh fruiting bodies of the edible mushroom Hypsizigus marmoreus. AB - An 18-kDa ribonuclease (RNase) with a novel N-terminal sequence was purified from fresh fruiting bodies of the mushroom Hypsizigus marmoreus. The purification protocol comprised ion exchange chromatography on DEAE cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-cellulose and Q-Sepharose and gel filtration by fast protein liquid chromatography on Superdex 75. The starting buffer was 10 mM Tris-HCl buffer (pH 7.2), 10 mM Tris HCl buffer (pH 7.2), 10 mM NH(4)OAc buffer (pH 5), 10 mM NH(4)HCO(3) buffer (pH 9.4) and 200 mM NH(4)HCO(3) (pH 8.5), respectively. Absorbed proteins were desorbed using NaCl added to the starting buffer. A 42-fold purification of the enzyme was achieved. The RNase was unadsorbed on DEAE cellulose, Affi-gel blue gel and CM-cellulose but adsorbed on Q-Sepharose. It exhibited maximal RNase activity at pH 5 and 70 degrees C. Some RNase activity was detectable at 100 degrees C. It demonstrated the highest ribonucleolytic activity (196 U/mg) toward poly C, the next highest activity (126 U/mg) toward poly A, and much weaker activity toward poly U (48 U/mg) and poly G (41 U/mg). The RNase inhibited [(3)H methyl]-thymidine uptake by leukemia L1210 cells with an IC(50) of 60 microM. PMID- 17920202 TI - Amnion in the treatment of pediatric partial-thickness facial burns. AB - BACKGROUND: Wound coverage for second-degree burns remains a clinical challenge. Human amniotic membranes have been used for many years in the treatment of burns; however, no large prospective clinical trials have been published. In this article, we present a novel and standardized procurement and processing method for amnion and investigate, whether the use of this biological dressing is safe and may represent a new therapeutic option for children with partial-thickness facial burns compared to standard topical treatment. METHODS: Patients with partial-thickness burns of the face, neck and head admitted between 2003 and 2005 were included in this study. They were divided into two groups to receive either amnion (n=53) or topical antimicrobials (n=49). Demographics (age, gender, ethnicity, TBSA, burn areas), length of hospital stay (LOS), rate of infections (RI), time to total healing, and frequency of dressing changes were compared between the two groups. The long-term outcome was assessed in nine patients in the amnion group and eight patients in the topical group, who returned for up to 12-month follow-up visits. RESULTS: Patients in the amnion group had significantly less dressing changes then in the control group (p<0.05). Time to healing, length of stay and the development of hypertrophic scarring was not different between the groups. Use of amnion was not associated with an increased risk of local infection. CONCLUSION: This study indicates that amnion is safe and has advantages as wound coverage for second-degree facial burns compared to the standard topical ointments. Further studies with the use of amniotic membranes on the trunk and the extremities, as well as for coverage of grafted third-degree burns, have yet to be performed. PMID- 17920203 TI - Base deficit and lactate: early predictors of morbidity and mortality in patients with burns. AB - Severe burn results in severe and unique physiological changes called burn shock. Historically, resuscitation has been guided by a combination of basic laboratory values, invasive monitoring and clinical findings, but the optimal guide to the endpoint of resuscitation still remains controversial. Two hundred and eighty patients, who were admitted to our Burn Unit, were enrolled in this prospective study. Resuscitation of these patients was undertaken according to the current standard of care. Parkland formula was used as a first approximation of acquired fluid administration rates; final fluid administration was adapted in order to meet clinical needs. The aim of this study was to evaluate if plasma lactate (PL) and base deficit (BD) are useful early parameters to estimate the severity of a burn. One of the main objectives was to evaluate if BD and its changes due to fluid resuscitation adds additional information in comparison to the evaluation of PL alone. The results of this study indicate that initial PL and BD level (Day 0) are useful parameters to separate survivors from non-survivors. Moreover, an outcome predictor of shock and effective resuscitation could be defined by evaluating the changes of BD on Day 1. Normalization of the BD within 24 h is associated with a better chance of survival. One explanation for this phenomenon might be the fact that many burn patients are still sub-optimally resuscitated; in summary, measuring PL and BD may help to identify critically injured patients either for enhancement of treatment, or selection of therapeutic options. PMID- 17920204 TI - The suprachiasmatic nucleus functions beyond circadian rhythm generation. AB - We recently discovered that human activity possesses a complex temporal organization characterized by scale-invariant/self-similar fluctuations from seconds to approximately 4 h-(statistical properties of fluctuations remain the same at different time scales). Here, we show that scale-invariant activity patterns are essentially identical in humans and rats, and exist for up to approximately 24 h: six-times longer than previously reported. Theoretically, such scale-invariant patterns can be produced by a neural network of interacting control nodes-system components with feedback loops-operating at different time scales. However such control nodes have not yet been identified in any neurophysiological model of scale invariance/self-similarity in mammals. Here we demonstrate that the endogenous circadian pacemaker (suprachiasmatic nucleus; SCN), known to modulate locomotor activity with a periodicity of approximately 24 h, also acts as a major neural control node responsible for the generation of scale-invariant locomotor patterns over a broad range of time scales from minutes to at least 24 h (rather than solely at approximately 24 h). Remarkably, we found that SCN lesion in rats completely abolished the scale-invariant locomotor patterns between 4 and 24 h and significantly altered the patterns at time scales <4 h. Identification of the control nodes of a neural network responsible for scale invariance is the critical first step in understanding the neurophysiological origin of scale invariance/self-similarity. PMID- 17920205 TI - Mesencephalic dopamine neuron number and tyrosine hydroxylase content: Genetic control and candidate genes. AB - The mesotelencephalic dopamine system shows substantial genetic variation which fundamentally affects normal and pathological behaviors related to motor function, motivation, and learning. Our earlier radioenzyme assay studies demonstrated significantly higher activity of tyrosine hydroxylase (TH), the first and rate limiting enzyme in the biosynthesis of catecholamine neurotransmitters, in the substantia nigra-ventral tegmental area of BALB/cJ mice in comparison with that of C57BL/6ByJ mice. Here, using quantitative immunoblotting and immunocytochemistry, we tested the hypothesis that mesencephalic TH protein content and number of nigral TH-positive neurons show strain-dependent differences in C57BL/6ByJ and BALB/cJ parallel to those observed in the TH activity studies. Immunoblotting experiments detected significantly higher mesencephalic TH protein content in BALB/cJ in comparison to C57BL/6ByJ (P<0.05). Immunocytochemical studies demonstrated that the number of TH-positive cells in substantia nigra was 31.3% higher in BALB/cJ than that in C57BL/6ByJ (P<0.01), while the average dopamine neuron volume was not significantly different. In a search for candidate genes that modulate TH content and the size of mesencephalic dopamine neuron populations we also studied near-isogenic mouse sublines derived from the C57BL/6ByJ and BALB/cJ progenitor strains. A whole genome scan with 768 single nucleotide polymorphism markers indicated that two sublines, C4A6/N and C4A6/B, were genetically very similar (98.3%). We found significantly higher mesencephalic TH protein content in C4A6/B in comparison to C4A6/N (P=0.01), and a tendency for higher number of dopamine neurons in the substantia nigra in C4A6/B in comparison to C4A6/N, which, however, did not reach statistical significance. To identify the genetic source of the TH content difference we analyzed the single nucleotide polymorphism (SNP) genotype data of the whole-genome scan, and detected two small differential chromosome segments on chr. 13 and chr. 14. Microarray gene expression studies and bioinformatic analysis of the two differential regions implicated two cis-regulated genes (Spock1 and Cxcl14, chr. 13), and two growth factor genes [bone morphogenetic protein 6 (Bmp6) (chr. 13), and fibroblast growth factor 14 (Fgf14) (chr. 14)]. Taken together, the results suggest that (1) nigral dopamine neuron number and TH protein content may be genetically associated but further studies are needed to establish unequivocally this linkage, and (2) Spock1, Cxcl14, Bmp6, and Fgf14 are novel candidates for modulating the expression and maintenance of TH content in mesencephalic dopamine neurons in vivo. PMID- 17920206 TI - Neural mechanisms of pelvic organ cross-sensitization. AB - Clinical observations of viscerovisceral referred pain in patients with gastrointestinal and genitourinary disorders suggest an overlap of neurohumoral mechanisms underlying both bowel and urinary bladder dysfunctions. Close proximity of visceral organs within the abdominal cavity complicates identification of the exact source of chronic pelvic pain, where it originates, and how it relocates with time. Cross-sensitization among pelvic structures may contribute to chronic pelvic pain of unknown etiology and involves convergent neural pathways of noxious stimulus transmission from two or more organs. Convergence of sensory information from discrete pelvic structures occurs at different levels of nervous system hierarchy including dorsal root ganglia, the spinal cord and the brain. The cell bodies of sensory neurons projecting to the colon, urinary bladder and male/female reproductive organs express a wide range of membrane receptors and synthesize many neurotransmitters and regulatory peptides. These substances are released from nerve terminals following enhanced neuronal excitability and may lead to the occurrence of neurogenic inflammation in the pelvis. Multiple factors including inflammation, nerve injury, ischemia, peripheral hyperalgesia, metabolic disorders and other pathological conditions dramatically alter the function of directly affected pelvic structures as well as organs located next to a damaged domain. Defining precise mechanisms of viscerovisceral cross-sensitization would have implications for the development of effective pharmacological therapies for the treatment of functional disorders with chronic pelvic pain such as irritable bowel syndrome and painful bladder syndrome. The complexity of overlapping neural pathways and possible mechanisms underlying pelvic organ crosstalk are analyzed in this review at both systemic and cellular levels. PMID- 17920207 TI - Vicious cycle composed of gut flora and visceral fat: a novel explanation of the initiation and progression of atherosclerosis. AB - Microorganisms infection was thought to be associated with atherosclerosis (AS), but the results of trials investigating antibiotic therapy in patients with coronary artery disease were controversial. Recently, a series of studies explored the relationship between gut flora and obesity, which showed that western-style diet (high fat) could induce imbalanced ratio of the Firmicutes versus the Bacteroidetes in the gut of germ-free mice and human beings, and gut flora could promote obesity through several novel pathways, such as inhibition of fasting-induced adipocyte factor. Meanwhile, data from some studies confirm that some link exists between the abdominal obesity and satiety centers (hindbrain and hypothalamus) with neurotransmitters and hormones. All of the above showed some connection between western-style diet, gut flora, visceral fat, and satiety centers, but until now, no study has shown us the exact mechanism about it. It is proposed that the vicious cycle composed of gut flora and visceral fat may initiate and promote AS. Attempts to confirm this hypothesis may lead to new directions in the study of the pathogenesis of AS and the development of novel strategies for the treatment of AS. PMID- 17920208 TI - Autism and vitamin D. AB - Any theory of autism's etiology must take into account its strong genetic basis while explaining its striking epidemiology. The apparent increase in the prevalence of autism over the last 20 years corresponds with increasing medical advice to avoid the sun, advice that has probably lowered vitamin D levels and would theoretically greatly lower activated vitamin D (calcitriol) levels in developing brains. Animal data has repeatedly shown that severe vitamin D deficiency during gestation dysregulates dozens of proteins involved in brain development and leads to rat pups with increased brain size and enlarged ventricles, abnormalities similar to those found in autistic children. Children with the Williams Syndrome, who can have greatly elevated calcitriol levels in early infancy, usually have phenotypes that are the opposite of autism. Children with vitamin D deficient rickets have several autistic markers that apparently disappear with high-dose vitamin D treatment. Estrogen and testosterone have very different effects on calcitriol's metabolism, differences that may explain the striking male/female sex ratios in autism. Calcitriol down-regulates production of inflammatory cytokines in the brain, cytokines that have been associated with autism. Consumption of vitamin D containing fish during pregnancy reduces autistic symptoms in offspring. Autism is more common in areas of impaired UVB penetration such as poleward latitudes, urban areas, areas with high air pollution, and areas of high precipitation. Autism is more common in dark-skinned persons and severe maternal vitamin D deficiency is exceptionally common the dark skinned. CONCLUSION: simple Gaussian distributions of the enzyme that activates neural calcitriol combined with widespread gestational and/or early childhood vitamin D deficiency may explain both the genetics and epidemiology of autism. If so, much of the disease is iatrogenic, brought on by medical advice to avoid the sun. Several types of studies could easily test the theory. PMID- 17920209 TI - Psychopathology in the context of obesity: the adiponectin hypothesis. PMID- 17920210 TI - Expression of two testis-specific genes, TSGA10 and SYCP3, in different cancers regarding to their pathological features. AB - BACKGROUND: Cancer-testis genes are a group of genes expressed in testicular germinal cells and a range of human cancers. Testis-specific gene A10 (TSGA10) is expressed in testis and actively dividing and fetal differentiating tissues. Mouse homologue (Tsga10) mRNA is translated to a 65kDa protein and appears to be processed to a major fibrous sheath protein of sperm tail. SYCP3 gene is supposed to be a testis-specific gene and constitutes the core of the lateral elements of synaptonemal complex. It has role in regulating DNA binding to the chromatid axis, sister chromatid cohesion, synapsis, and recombination. METHODS: In this study expression of TSGA10 and SYCP3 were investigated in different cancers (156 tumor samples) using RT-PCR. Diagnosis of cancer was based on histopathological reports. The association with histopathological characteristics of tumors was analyzed using statistical programs. RESULTS: TSGA10 expression was observed in 83% of brain tumors, 66% of breast cancers, 58% of gastrointestinal tumors, 66% of skin tumors and 53% of soft tissue tumors. But, SYCP3 transcripts were found in four tumor samples (moderately differentiated gemistocytic astrocytoma, pituitary adenoma, glioma and an ovarian tumor). CONCLUSION: These results may get further insight into TSGA10, but not SYCP3, potential role as a cancer marker and a cancer testis gene implicated in tumorogenesis of cancers. PMID- 17920211 TI - Molecular and serological diagnosis of Chikungunya virus infection. AB - INTRODUCTION: In 2005-2006, during the Chikungunya virus outbreak in La Reunion (Indian Ocean), we urgently established the molecular and serological methods for the diagnosis of Chikungunya virus (CHIKV) from various types of samples. METHODS: CHIKV RNA was detected using a highly sensitive real-time RT PCR assay. A co-extracted and co-amplified internal control RNA was used to identify RT PCR inhibitors. Depending on their nature samples were pretreated before nucleic acid extraction. Viral loads were measured using a synthetic RNA calibrator. CHIKV immunoglobulin (Ig) G and M antibodies were detected by ELISA either from sera or from blood absorbed on filter paper. RESULTS: CHIKV RNA was found in various types of samples such as plasma, cerebrospinal fluid, and placenta, but was not found in some samples including maternal milk and synovial samples. Detection of IgG from filter paper absorbed blood is specific and sensitive. Routine data showed that maternally transferred IgG and naturally acquired IgM persist at least 12 and 18 months, respectively. DISCUSSION: The techniques enabled the diagnosis of chikungunya in known and newly described forms of the disease. They are used for routine diagnosis and large scale surveys. PMID- 17920212 TI - Mutational and biochemical analyses of the endolysin Lys(gaY) encoded by the Lactobacillus gasseri JCM 1131T phage phi gaY. AB - The Lys(gaY) of Lactobacillus gasseri JCM 1131(T) phage phigaY endolysin was purified to homogeneity using the Escherichia coli/His.Tag system. Zymographic and spectrophotometric assays showed that Lys(gaY) lysed over 20 heated Gram positive bacterial species as the substrates, including lactobacilli, lactococci, enterococci, micrococci, and staphylococci. The enzymatic activity had the pH and temperature optima of about 6.5 and 37 degrees C, respectively. Amino-acid substitution analysis revealed that 13 residues of Lys(gaY) were involved in cell lytic activity: in the beta/alpha(gaY) domain, G10, D12, E33, D36, H60, Y61, D96, E98, V124, L132, and D198; in the SH3b(gaY) domain, Y272 and W284. In addition, deletion analysis demonstrated that the beta/alpha(gaY) domain of N-terminal 216 residues is the core enzyme portion, although the cell-lytic ability is lower than that of Lys(gaY). These mutational experiments suggested that beta/alpha(gaY) (in which two acidic residues of D12 and E98 likely act as catalytic residues) is responsible for cell-lytic activity, and SH3b(gaY) promotes beta/alpha(gaY) possibly through cell-wall binding function. The purified His-tagged SH3b(gaY) domain containing 94 residues from S217 to K310 (i) bound to Gram-positive bacteria susceptible to Lys(gaY), (ii) induced aggregation of exponentially growing cells of L. gasseri JCM 1131(T), L. casei IAM 1045, Lactococcus lactis C2, L. lactis MG 1363, and Enterococcus hirae IAM 1262 by forming thread-like chained cells, (iii) inhibited lytic activity of Lys(gaY), and (iv) impeded autolysis of L. gasseri JCM 1131(T) in buffer systems. A truncated protein HDeltaSH3b(gaY) lacking in N-terminal 21 residues (from S217 to E237) of SH3b(gaY) and an amino-acid substituted protein HSH3b(gaY)G (W284G) lost the activities of HSH3b(gaY), showing that the N-terminal region and W284 probably play important roles in the SH3b(gaY) function(s). PMID- 17920213 TI - The immunogenicity of fusion protein linking the carboxyl terminus of the B subunit of Shiga toxin 2 to the B subunit of E. coli heat-labile enterotoxin. AB - To augment the immunogenicity of the subunit B of Shiga toxin (Stx2e B) produced by Escherichia coli and protect piglets from edema disease in china, a fusion gene was constructed consisting of Stx2e B genetically linked at the N-terminus of the B subunit of heat-labile enterotoxin (LTB) in a translational fusion. After being induced with IPTG, the expressed fusion protein of Stx2e B-LTB was about 8.8% of total proteins, approximately 13 microg/ml of the bacteria culture. The Stx2e B-LTB fusion protein was found to be nontoxic to Vero cells at the dose higher than 1 microg/ml and to mice less than 100 microg/ml. Antibody titer against the fusion protein Stx2e B-LTB was 1:76,800, much higher than that of the recombinant Stx2e B protein (1:12,800) alone. All of the mice immunized with the Stx2e B-LTB fusion protein survived when challenged with a lethal dose (LD) of Stx2e toxin. The results showed that the poor immunogenicity of Stx2e B was overcome by conjugating the stx2e B to ltB. The immunogenicity of the constructed fusion protein Stx2e B-LTB in the present study was highly qualified to protect animals against Shiga toxin produced from Shiga toxin-producing Escherichia coli (STEC). The fusion protein of Stx2e B-LTB could be a candidate for a vaccine against edema disease and post-weaning diarrhea simultaneously in piglets. PMID- 17920214 TI - Fatty acids and early human development. AB - Fatty acids play central roles in growth and development through their roles in membrane lipids, as ligands for receptors and transcription factors that regulate gene expression, precursor for eicosanoids, in cellular communication, and through direct interactions with proteins. Adverse fatty acid supplies during fetal and child development alter the fatty acid composition of membrane phospholipids and storage triglycerides with the potential to disrupt cellular environments, and program structure and function. Maternal fatty acid nutrition during pregnancy and lactation determines the transfer of essential n-6 and n-3, and non-essential trans fatty acids via the placenta and through human milk. Poor maternal docosahexaenoic acid (DHA) status increases risk of inadequate DHA to support brain and retinal development, delaying or limiting neural and visual system development. The implications of recent changes in the dietary fatty acids on maternal to infant fatty acid transfer, including the composition of human milk has been insufficiently studied. PMID- 17920215 TI - Hormone therapy use and prescription durations of menopausal women in Taiwan: a 5 years' National Cohort study. AB - OBJECTIVES: To describe the prevalence and associated factors of hormone replacement therapy (HRT)-related utilization in a national representative fixed cohort in Taiwan. METHOD: The study population are women aged 40 years and over in Taiwan. Our data, provided by National Health Research Institutes, are the sampled registry information of 37,315 beneficiaries of National Health Insurance from 2000 to 2004. The dependent variables were HRT-related utilizations, including physician contact, HRT utilization rate/duration/patterns (new, prevalent and discontinue users). The independent variables were age, time, prior co-morbidities and prior utilization pattern. We used generalized estimating equation (GEE) model for repeated measurement analysis. RESULTS: The outpatient contact rates for menopause syndrome were low, though the HRT prescription rate among those who have contact were high. GEE shows that age, time, prior co morbidities/HRT utilization patterns were significantly associated with all types of HRT-related utilizations, which all declined significantly following the publications of Women's Health Initiative (WHI) in 2002. The magnitudes of reduction, though similar in 2003-2004, were greater for physician contact and HRT durations than that of HRT prescription rate for those have contacts in 2002. Besides, the percentages of new (discontinue) users have already declined (increased) since 2001 although it had not declined until 2003 for the continued users. CONCLUSIONS: WHI publications have great impact on HRT-related utilizations. However, the response of the women was quicker and much drastic than that of the physicians in 2002. Besides, the efforts of the various women's associations before WHI might have some contribution to the declined (increased) of new (discontinued) users. PMID- 17920216 TI - Interaction of lipid nanoparticles with human epidermis and an organotypic cell culture model. AB - Various lipid nanoparticle formulations were investigated with respect to (trans)dermal drug delivery with special regard to the mechanism of their effects on human and an organotypic cell culture epidermis. Potential alterations of stratum corneum lipid domains were studied using fluorescence assays with labeled liposomes and thermal analysis of isolated stratum corneum. Influences on the permeation of corticosterone were investigated and the occlusive properties of the nanoparticles were determined by measurements of the transepidermal water loss (TEWL). The penetration of a fluorescence dye was visualized by fluorescence microscopy of cross sections of human epidermis after incubation with cubic and solid lipid nanoparticles. Corticosterone permeation was limited when applied in matrix-type lipid nanoparticles (fat emulsion, smectic and solid lipid nanoparticles). An adhesion of solid lipid nanoparticles was clearly observed in thermal analysis as reflected by additional phase transitions probably caused by the nanoparticle matrix lipid. However, as for the other matrix-type nanoparticles, no distinct alterations of the phase transitions of the stratum corneum lipids were observed. Cubic nanoparticles led to the most predominant effect on skin permeation where the surface-active matrix lipid may act as penetration enhancer. An alteration of the stratum corneum lipids' thermal behavior as well as an interaction with fluorescence labeled liposomes was observed. Differences observed in permeation studies and thermal analysis of human and cell culture epidermis indicate that surface lipids, which are not present to the same extent in the cell culture model than in human epidermis, seem to play an important role. PMID- 17920217 TI - Influence of sulfobutyl ether beta-cyclodextrin (Captisol) on the dissolution properties of a poorly soluble drug from extrudates prepared by hot-melt extrusion. AB - The aim of this study was to investigate the influence of sulfobutyl ether beta cyclodextrin (SBE(7)-beta-CD; Captisol on the dissolution properties of a poorly water-soluble drug from extrudates prepared by hot-melt extrusion. Ketoprofen was employed as a model drug. Extrudates containing the parent beta-cyclodextrin (beta-CD) were also produced for comparative evaluation to assess the benefits of SBE(7)-beta-CD. Hot-melt extrudates were produced at 100 degrees C, which was close to the melting point of ketoprofen. The physiochemical properties and the in vitro drug release properties of ketoprofen from extrudates were investigated and compared with samples prepared by physical mixing, co-grinding, freeze-drying and heat-treatment. The solubilizing effects and the interactions of ketoprofen with SBE(7)-beta-CD and beta-CD were investigated using phase solubility and NMR studies, respectively. The dissolution rate of ketoprofen from samples prepared by hot-melt extrusion with SBE(7)-beta-CD was significantly faster than both the physical mixture and the hot-melt extrudates prepared with the parent beta-CD. Moisture absorption studies revealed that the hygroscopic nature of SBE(7)-beta CD led to particle aggregation and a corresponding decrease in drug release rate for all samples. However, the samples prepared by melt extrusion were least affected by exposure to elevated humidity. PMID- 17920218 TI - d-alpha-Tocopheryl polyethylene glycol 1000 succinate (TPGS) modified poly(l lactide) (PLLA) films for localized delivery of paclitaxel. AB - d-alpha-Tocopheryl polyethylene glycol 1000 succinate (TPGS) was used as a novel additive to the poly(l-lactide) (PLLA) films for local drug delivery with paclitaxel as a prototype therapeutic agent. Paclitaxel-loaded PLLA/TPGS films were prepared by the solvent casting technique with dichloromethane as the solvent. Effects of TPGS component on the films' physicomechanical properties and the drug release profile were investigated. It was found by field emission scanning microscopy (FESEM) that a biphasic honeycomb surface was formed for the PLLA/TPGS films, while the PLLA film exhibited a smooth and homogeneous surface. There was no significant effect of the drug loading on the morphological structure of the PLLA/TPGS films. Differential scanning calorimetry (DSC) demonstrated that the PLLA/TPGS films was a phase-separated system. Tensile testing showed that the flexibility of the PLLA/TPGS films was much higher than that of the PLLA film. The elongation at break for the PLLA/TPGS film of 5%, 10% and 15% TPGS content was 6.8, 8.9 and 19.4 times of that for the PLLA film, respectively. In vitro drug release studies found that incorporation of TPGS considerably facilitated paclitaxel release. PMID- 17920220 TI - Ictal electroencephalographic findings of neonatal seizures in preterm infants. AB - The aim of this study is to clarify the characteristics of ictal EEG findings of neonatal seizures in preterm infants. Seizures associated with ictal EEG changes were recognized in nine infants with gestational age of less than 37 weeks. Propagation, migration, shifting, changes in morphology of ictal EEG discharges were evaluated. Seizure manifestation was divided into the following categories; motor seizure, apneic seizure, automatic seizure and seizure without clinical symptoms. The types of the seizures were motor seizures in five infants, apneic in two, automatic in one and those without clinical symptoms in five. All seizures were of focal onset. The foci of seizures were temporal in six infants, occipital in two, central in one, and frontal in one. The morphology of ictal discharges was low voltage spikes or sharp waves in six infants, spikes in two, theta waves in one and high-voltage spiky theta in one. The propagation of ictal discharges was focal in five infants and regional in five. The migration of ictal discharges was observed in two infants and a shift in two. There was no clear relation between seizure manifestation and ictal EEG foci, duration of seizures and morphology or propagation of ictal discharges. PMID- 17920219 TI - Anger symptoms and "delinquent" behavior in Tourette syndrome with and without attention deficit hyperactivity disorder. PMID- 17920221 TI - [Initial training in vascular medicine: a revision was needed!]. PMID- 17920222 TI - Quantitative assessment of the influence of anatomic noise on the detection of subtle lung nodule in digital chest radiography using fractal-feature distance. AB - PURPOSE: To confirm whether or not the influence of anatomic noise on the detection of nodules in digital chest radiography can be evaluated by the fractal feature distance. MATERIALS AND METHODS: We used the square images with and without a simulated nodule which were generated in our previous observer performance study; the simulated nodule was located on the upper margin of a rib, the inside of a rib, the lower margin of a rib, or the central region between two adjoining ribs. For the square chest images, fractal analysis was conducted using the virtual volume method. The fractal-feature distances between the considered and the reference images were calculated using the pseudo-fractal dimension and complexity, and the square images without the simulated nodule were employed as the reference images. We compared the fractal-feature distances with the observer's confidence level regarding the presence of a nodule in plain chest radiograph. RESULTS: For all square chest images, the relationships between the length of the square boxes and the mean of the virtual volumes were linear on a log-log scale. For all types of the simulated nodules, the fractal-feature distance was the highest for the simulated nodules located on the central region between two adjoining ribs and was the lowest for those located in the inside of a rib. The fractal-feature distance showed a linear relation to an observer's confidence level. CONCLUSION: The fractal-feature distance would be useful for evaluating the influence of anatomic noise on the detection of nodules in digital chest radiography. PMID- 17920223 TI - Magnetic resonance spectroscopy features of normal-appearing white matter in patients with acute brucellosis. AB - We aimed to evaluate whether the subtle metabolic cerebral changes are present in normal-appearing white matter on conventional MRI, in patients with acute brucellosis, by using MR spectroscopy (MRS). Sixteen patients with acute brucellosis and 13 healthy control subjects were investigated with conventional MRI and single-voxel MRS. Voxels were placed in normal-appearing parietal white matter (NAPWM). N-Acetyl aspartate (NAA)/creatine (Cr) and choline (Cho)/Cr ratios were calculated. There was no significant difference between the study subjects and the control group in NAA/Cr ratios obtained from NAPWM. However, the Cho/Cr ratios were significantly higher in patients with acute brucellosis compared to controls (p=0.01). MRS revealed metabolic changes in normal-appearing white matter of patients with brucellosis. Brucellosis may cause subtle cerebral alterations, which may only be discernible with MRS. Increased Cho/Cr ratio possibly represents an initial phase of inflammation and/or demyelination process of brucellosis. PMID- 17920224 TI - The forgotten organ: contrast enhanced sonography of the spleen. AB - OBJECTIVE: Ultrasound contrast agents in conjunction with contrast specific imaging techniques, are increasingly accepted in clinical use for diagnostic imaging in several organs. Contrast enhanced sonography (CES) of second generation contrast media have shown a spleen-specific uptake of the microbubble contrast agent. The aim of this review is to illustrate indications for the use of CES in patients with suspected (peri-)splenic pathology. METHODS: This review based on the experience of transcutaneous CES in 200 patients with (peri-)splenic pathology diagnosed by B-mode sonography at an internal medicine center. CES studies were performed with a contrast-devoted unit (Acuson, Sequoia, Siemens medical solution) that had contrast-specific, continuous-mode software. A low mechanical index was used. A sulfur hexafluoride-based microbubble contrast medium (Sonovue, Bracco SpA, Milan, Italy) was injected. RESULTS: On our experience, there are several clinical conditions which may show an diagnostic advantage of CES in comparison to B-mode US. CES should be performed to investigate: (1) the perisplenic tumor to diagnose or exclude accessory spleen, (2) the small-sized spleen to diagnose functional asplenia/hyposplenia, (3) the inhomogenous spleen of unknown cause to diagnose focal lesions within the spleen, (4) the incidentally found hypoechoic splenic tumor to diagnose high vascular splenic hemangioma, (5) focal lesions suspect for splenic abscess, hematoma, infarction to confirme diagnosis, and (6) patients with abdominal trauma to diagnose or exclude splenic injury. CONCLUSION: CES is of diagnostic value in several clinical circumstances to diagnose accessory spleen, functional asplenia, small-sized splenic involvement, high vascular splenic hemangioma, and vascular splenic pathology like splenic infarction, splenic abscess, and splenic laceration. PMID- 17920225 TI - Using of HPLC coupled with coulometric detector for the determination of biotin in pharmaceuticals. AB - The method for the determination of biotin by high performance liquid chromatography (HPLC) coupled with coulometric detector is presented here. Chromatographic and detection conditions were tested. A LiChrospher 60RP-select B column (250 mm x 4 mm; 5 microm) and the mobile phase containing 0.24 mol/L aqueous solution of acetic acid and acetonitrile in the ratio 85:15 (v/v) were found as the most suitable. The flow rate was 1 mL/min and the injected volume of the sample was 20 microL. The hydrodynamic voltammogram of biotin was measured and according to obtained data the detection parameters were set--channel I 600 mV, channel II 900 mV, sensitivity 1 microA. The developed method has been validated. The calibration curve is linear in the range 15-3600 ng/mL, correlation coefficient is 0.9998, limits of detection and quantification are 5 and 15 ng/mL, respectively. Recovery of the spiked samples was 98.67% with R.S.D. 0.255% on average. The developed method has been successfully applied for determination of biotin in pharmaceutical preparations. PMID- 17920226 TI - Untangling the Web--the impact of Internet use on health care and the physician patient relationship. AB - OBJECTIVE: The use of Web (i.e. Internet)-derived health information within the health care encounter is rapidly increasing. In this article, an extensive review of the complex effects and sometimes contradictory roles of the Web in regard to health care delivery and the physician-patient relationship is presented. METHODS: A review of relevant literature was conducted, with key points integrated into a physician guide for effective interaction with Web-activated patients. RESULTS: An emerging consumerist model with "triangulation" of patient Web-physician can be expected to significantly impact dynamics of the physician patient relationship. Potential advantages of Web-acquired information include helping patients make informed health care choices (with potential to decrease health care disparities), shared decision-making with a collaborative, teamwork approach, more efficient use of clinical time, augmenting of physician-provided information, online support groups, and/or access to patients' own health information. Alternatively, factors such as misinformation due to highly variable quality of Web information, possible exacerbation of socioeconomic health disparities, and shifting of conventional notions of the physician-patient relationship ("traditional" medical authority) present their own set of challenges for the health care provider. CONCLUSION: A tangible guide to the integration of patients' use of the Web within a medical practice is thus offered with recommended communication skills. PRACTICE IMPLICATIONS: The "net-friendly" clinician can be effective by engendering a genuine partnership with patients, thus contributing to quality health care. PMID- 17920227 TI - The relative importance of graft surveillance and warfarin therapy in infrainguinal prosthetic bypass failure. AB - BACKGROUND: We sought to describe modes of failure and associated limb loss after infrainguinal polytetrafluoroethylene bypass grafting in patients lacking a saphenous venous conduit and to define specific clinical or hemodynamic factors prognostic for bypass failure. METHODS: We identified 121 patients (mean age, 67 years; 90 men and 31 women) with determinable outcomes (minimum follow-up, 2 months; mean, 17 months) after 130 prosthetic infrainguinal bypasses between 1997 and 2005. Ischemic presentation was rest pain in 52%, tissue loss in 34%, and disabling claudication and/or popliteal aneurysm in 14%, with 24% of patients requiring a redo bypass. Distal targets were the above-knee (n = 44), distal popliteal (n = 27), or tibial/pedal (n = 59) arteries. Sixty-six (77%) of the below-knee (BK) target (distal popliteal or tibial) bypasses had distal anastomotic adjuncts (vein cuff or patch). Duplex graft surveillance was performed at 1, 4, and 7 months after surgery and twice yearly thereafter, with recording of midgraft velocities and imaging encompassing inflow and outflow vessels. Arteriography and open/endovascular intervention was performed for stenoses identified by duplex scanning (peak systolic velocity >300 cm/s; velocity ratio >3.5). An attempt was made to salvage occluded grafts by using catheter-directed thrombolysis or open techniques. Eighty-six patients (74% of BK bypasses) were placed on chronic warfarin therapy with a target international normalized ratio range between 2 and 3. Prognostic factors were identified by using univariate statistics and multivariate logistic regression analysis. RESULTS: Three-year primary, assisted, and secondary patency rates were 39%, 43%, and 59%, respectively, for all bypasses, with no difference noted between above knee and BK grafts (P = .5). At 3 years, freedom from limb loss was 75%, and patient survival was only 70%, with no adverse effect on survival imparted by amputation. Sixty-nine total adverse events occurred as a result of thrombotic occlusion (n = 51), duplex scan-detected stenosis (n = 13), or graft infection (n = 5). Forty-nine percent of all initial graft occlusions eventually led to amputation. Twenty-three grafts (27% of 86 patients) in patients maintained on chronic warfarin were subtherapeutic at the time of occlusion. Use of a distal anastomotic adjunct with BK bypasses reduced graft thrombosis (35% with vs 60% without) but did not impart a significant patency advantage (P = .07). Multivariate analysis revealed low graft flow (midgraft velocity < or =45 cm/s; odds ratio [OR], 6.1; 95% confidence interval [CI], 1.9-19.2), use of warfarin (OR, 8.4; 95% CI, 2.1-34.5), and therapeutic warfarin (OR, 24.6; 95% CI, 5.7-106) to be independently predictive for bypass patency. Graft patency was maintained in 89% of grafts remaining therapeutic on warfarin compared with only 55% of subtherapeutic or nonanticoagulated grafts (P < .001). Low-flow grafts (n = 61) occluded more frequently than higher-flow grafts (46% vs 13%; P < .001). Therapeutic warfarin augmented the patency of low-flow (P < .001) but not high flow (P = .15) grafts. CONCLUSIONS: Low graft flow was a more common mode of prosthetic bypass failure than development of duplex scan-detected stenotic lesions during follow-up. Early duplex scanning may be more important for characterizing midgraft velocity and related thrombotic potential and selecting patients for chronic anticoagulation. Maintenance of therapeutic warfarin is paramount in optimizing prosthetic bypass patency and limb preservation. PMID- 17920228 TI - The use of intravascular ultrasound imaging to improve use of inferior vena cava filters in a high-risk bariatric population. AB - OBJECTIVE: Pulmonary embolism is the leading cause of death after gastric bypass procedures for obesity, approximating 0.5% to 4%. All bariatric patients, but especially the super-obese, which have a body mass index (BMI) >50 kg/m(2), are at significant risk for postoperative venous thromboembolism (VTE). Visualization and weight limitations of fluoroscopy tables exclude most bariatric and all super obese patients from inferior vena cava (IVC) filter placement using fluoroscopy. Intravascular ultrasound (IVUS)-guided IVC filter placement is the only modality that allows these high-risk patients to have an IVC filter placed. METHODS: Hospital and outpatient records of the 494 patients who underwent gastric bypass procedures from January 1, 2004, to May 31, 2006, were reviewed. All patients who had concurrent IVC filter placement with the use of IVUS guidance were selected. Comorbidities, outcomes, and complications were recorded. RESULTS: We identified 27 patients with mean BMI of 70 +/- 3 kg/m(2); of these, 25 were super-obese (BMI >50 kg/m(2)). Procedures included five laparoscopic and 22 open gastric bypass operations. All patients underwent concurrent IVC filter placement using IVUS guidance. In addition to super-obesity, indications for IVC filter placement included history of VTE (n = 4), known hypercoagulable state (n = 2), and profound immobility (n = 21). Mean follow up was 293 +/- 40 days. Technical success rate was 96.3%. There were no catheter site complications. In one surviving patient, a nonfatal pulmonary embolism was detected by computed tomography 2 months postoperatively. Two patients died, and autopsy excluded VTE as the cause of death in both. CONCLUSION: This study suggests efficacy of IVUS guided IVC filter placement in preventing mortality from pulmonary embolism in high-risk bariatric patients, including the super-obese. IVUS-guided IVC filter placement can be safely performed with an excellent success rate in all bariatric patients, including the super-obese, who otherwise would not be candidates for IVC filter placement due to the limitations imposed by their large body habitus. PMID- 17920229 TI - Intraplaque hemorrhage assessed by high-resolution magnetic resonance imaging and C-reactive protein in carotid atherosclerosis. AB - BACKGROUND: Carotid intraplaque hemorrhage is a marker of atheroma instability. Noninvasive assessment of bleeding can be performed by high-resolution magnetic resonance imaging (MRI), but its association with inflammatory markers has not been clearly demonstrated. METHODS: We evaluated consecutive carotid endarterectomy patients that underwent high-resolution MRI, independent evaluation of neurologic symptoms, C-reactive protein measurement, and histologic analysis. Intraplaque hemorrhage was determined by the presence of a hyperintense MRI signal (T1-weighted sequence). RESULTS: The study included 70 predominantly male (66%) and hypertensive (89%) patients (89%) aged 66 +/- 9 years old. MR angiography identified 15 patients (21.5%) with stenosis between 50% and 69%, 15 (21.5%) with stenosis between 70% and 90%, and 40 (57%) with stenosis >90%. High resolution MRI depicted a hyperintense signal suggestive of intraplaque bleeding in 45 subjects (64%). All patients who had had transient ischemic attacks >90 days before the surgery showed a hyperintense signal on MRI (P = .007). Age, gender, traditional cardiovascular risk factors, and history of myocardial infarction or peripheral arterial disease were similar in patients with or without signs of intraplaque bleeding on MRI. There was excellent agreement between acute or recent hemorrhage on histologic and MRI findings (kappa coefficient, 0.91; 95% confidence interval, 0.81 to 1.00). Only one of 45 patients (2%) with a hyperintense signal on MRI did not have acute or recent hemorrhage in the histologic analysis (P < .001). High-sensitivity C-reactive protein levels were similar for different degrees of carotid stenosis as assessed by MR angiography, but they were significantly higher in clinically unstable patients (P = .006) and in those with a positive hyperintense MRI signal (P = .01). In an aggregated analysis of neurologic symptoms and MRI findings, we found a progressive increase of high-sensitivity C-reactive protein levels (P = .02). CONCLUSIONS: Intraplaque hemorrhage evaluated by MRI identified neurologically unstable patients with increased levels of high-sensitivity C-reactive protein regardless of the degree of carotid stenosis. PMID- 17920230 TI - The APC E1317Q and I1307K polymorphisms in non-colorectal cancers. AB - Mutation of the adenomatous polyposis coli (APC) gene is an important initiating factor in the early stages of the multi-step colorectal cancer (CRC) carcinogenesis. APC E1317Q and I1307K variants have been linked to CRC. The aim of this study was to examine the association of these variants with non colorectal cancers. Mutation screening was performed using real-time PCR. The APC E1317Q variant was detected in 1.25% individuals undergoing testing. Among 2076 patients that were analyzed for this mutation, 404 had cancer outside of the colon. None of the non-colorectal cancer patients was a carrier of the E1317Q polymorphism. The I1307K variant was found in 32 subjects with non-CRC (7.9%). We conclude herein that the E1317Q gene variant in the APC gene is not found in cancers outside of the colon. The prevalence of the more common I1307K variant is similar to that of CRC. PMID- 17920231 TI - Effect of Cervus korean TEMMINCK var. mantchuricus Swinhoe on protease activities, antioxidant and free radical damages in rheumatis arthritis rats. AB - The effect of deer antler extracts (DAA) of Cervus korean TEMMINCK var. mantchuricus Swinhoe on protease activities, oxidant and free radical damages in synovial fluid from rheumatoid arthritis in rats was studied. Rats were i.p. administered with DAA. We have compared (using the same series of experimental samples) the levels of activity of a comprehensive range of cytoplasmic, lysosomal and matrix protease types, together with the levels of free radical induced protein damage (determined as protein carbonyl derivative) and total antioxidant in synovial fluid from rheumatoid arthritis (RA) and DAA-treated rats. Many proteases activities were shown to be significantly increased in RA compared to normal rats. Protease activities (including those enzyme types putatively involved in the immune response, such as dipeptidyl aminopeptidase IV) in plasma were not significantly different between RA and normal rats. DAA treatment at dose of 100 microg/kg suppressed the production of the proteases of cytoplasmic, lysosomal and matrix protease types. The level of free radical induced damage to synovial fluid proteins was approximately 2-fold lower in DAA rats compared to RA rats, although there was no significant difference in total antioxidant status in synovial fluid or plasma between RA and DAA rats. It was concluded that DAA treatment reduces the activation of proteolytic enzymes and free radicals, which are likely to be of equal potential importance as protein damaging agents in the pathogenesis of RA. PMID- 17920233 TI - Combined treatment of P-gp-positive L1210/VCR cells by verapamil and all-trans retinoic acid induces down-regulation of P-glycoprotein expression and transport activity. AB - The development of the most common multidrug resistance (MDR) phenotype associated with a massive overexpression of P-glycoprotein (P-gp) in neoplastic cells may result in more than one hundred fold higher resistance of these cells to several drugs. L1210/VCR is a P-gp-positive drug resistant cell line in which P-gp overexpression was achieved by repeated cultivation of parental cells with a stepwise increasing concentration of vincristine. Relatively little is known about regulation of P-gp expression. Therefore, serious efforts have been made to recognize all aspects involved in regulation of P-gp expression. Retinoic acid nuclear receptors are involved in regulating expression of a large number of different proteins. Several authors have described that all-trans retinoic acid (ATRA, ligand of retinoic acid receptors, RARs) may induce alterations in P-gp expression and/or activity in drug resistant malignant cell lines. There are also other nuclear receptors for retinoids--retinoid X receptors (RXRs)--that may be involved in the development of the P-gp-mediated MDR phenotype. The topic of the present paper is a study of the relationship, if any, between the regulatory pathways of nuclear receptors for retinoids and P-glycoprotein expression. Increased levels of mRNAs encoding the retinoic acid nuclear receptors RARalpha and gamma, as well as decreased levels of the mRNAs encoding RARbeta and the retinoid X receptor RXRgamma or slightly decreased levels of RXRbeta mRNA, were observed in L1210/VCR cells in comparison with parental L1210 cells. Neither L1210 cells nor L1210/VCR cells contained measurable amounts of mRNA encoding the RXRalpha receptor. ATRA did not influence the viability of L1210/VCR cells differently from L1210 cells. A combined treatment of L1210/VCR cells with vincristine (1.08 micromol/l) and ATRA induced slightly higher cell death than that observed with ATRA alone. When applied alone, ATRA did not influence P-gp expression (monitored by anti P-gp antibody c219 using western blot analysis) or transport activity (monitored by use of calcein/AM as a P-gp substrate by FACS) in L1210/VCR cells. In contrast, when ATRA was applied together with verapamil (an often used P-gp inhibitor), a significant decrease in P-gp expression and transport activity were observed. However, no significant differences in [11, 12 (3)H]-ATRA uptake were observed in either sensitive or resistant cells, in the latter case in the absence or presence of vincristine. Moreover, verapamil did not influence ATRA uptake under any conditions. Thus, we can conclude that the combined treatment of L1210/VCR cells with ATRA and verapamil is able to depress P-gp expression, and consequently its activity. ATRA is not a P-gp-transportable substance, and thus this effect could not be attributed to verapamil-induced inhibition of P-gp that would allow ATRA to reach retinoic acid nuclear receptors and activate them. PMID- 17920232 TI - Anti-cancer activity of highly purified sulfur in immortalized and malignant human oral keratinocytes. AB - Sulfur is commonly used in Asia as an herbal medicine to treat inflammation and cancer, and potent chemopreventive effects have been demonstrated in various in vivo and in vitro models for sulfur-containing compounds found in naturally occurring products. Here, we report the growth inhibitory and apoptosis-related effects of a newly developed highly purified sulfur (HPS) on immortalized human oral keratinocytes (IHOKs) and on oral cancer cells representing two stages of oral cancer (HN4, HN12) based on a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay, Western blotting, cell cycle analysis, and nuclear staining. The purity of the sulfur preparation was verified by high performance liquid chromatography. HPS inhibited the proliferation of immortalized and malignant oral keratinocytes in a dose- and time-dependent manner. FITC-annexin V staining, DNA fragmentation testing, and Hoechst 33258 staining revealed that HPS inhibited cell growth via apoptosis. HPS increased the sub-G1 cell cycle fraction, with decreased expression of cyclins D1, D2, and E and their activating partners cdk2, cdk4, and cdk6, and a concomitant induction of p53 and p21/WAF1. Furthermore, HPS treatment increased the cytosolic level of cytochrome c and resulted in caspase-3 activation; this effect was correlated with Bax up-regulation and Bcl-2 down-regulation. Thus, these data suggest that HPS is a potential candidate for anti-cancer therapy in oral cancer. PMID- 17920234 TI - Cytotoxic effect of gallic acid on testicular cell lines with increasing H2O2 level in GC-1 spg cells. AB - Gallic acid is added to foods to prevent oxygen-induced lipid peroxidation and can be obtained by the hydrolysis of tannic acid which can be found in tea, coffee, red wine, and immature fruits. Tannic acid has also been used as a food additive. In the present study, we investigated the effect of gallic acid on mouse spermatogonia, mouse spermatocytes, and mouse Sertoli cells in vitro. First, each cell line was cultured with predetermined concentrations of gallic acid for 3h to access the effects of gallic acid on in vitro growth of testicular cells and MTT cytotoxicity assay was used to measure cell viability. Secondly, intracellular levels of hydrogen peroxide in mouse spermatogonia, mouse spermatocytes, and mouse Sertoli cells treated with gallic acid were analyzed using dihydrorhodamine 123 as a probe to evaluate the pro-oxidative property of gallic acid. The results obtained indicate that gallic acid inhibits the growth and proliferation of testicular cells in a dose-dependent manner and increases the intracellular level of hydrogen peroxide in mouse spermatogonia significantly (p<0.05). It can be suggested that gallic acid exerts cytotoxic effects on testicular cells by its pro-oxidative activity. In conclusion, gallic acid induced cytotoxicity in mouse spermatogonia, mouse spermatocytes, and mouse Sertoli cells in vitro may be of toxicological research interest considering the testicular toxic potential of gallic acid. PMID- 17920235 TI - Synergistic toxicity induced by prolonged glutathione depletion and inhibition of nuclear factor-kappaB signaling in liver cells. AB - TNF-alpha, GSH depletion and CYP2E1 are factors that play an important role in alcoholic liver disease. Activation of NF-kappaB prevents hepatocyte damage caused by TNF-alpha. This work describes the effect of NF-kappaB inhibition on toxicities caused by GSH depletion or arachidonic acid (AA) treatment in liver cells, and evaluates the possible influence of CYP2E1 overexpression. Cells were exposed to the NF-kappaB inhibitor BAY11-7082, in the absence or presence of l buthionine sulfoximine (BSO) to block GSH synthesis. BSO toxicity was higher in CYP2E1-expressing E47 HepG2 cells compared to control cells; the incubation with BAY11-7082 potentiated BSO toxicity in both cell lines comparably. Several other agents which suppress activation of NF-kappaB increased BSO toxicity in E47 cells. NF-kappaB inhibition, however, did not sensitize E47 cells to AA toxicity. Suppressing activity of NF-kappaB also potentiated BSO, but not AA toxicity, in isolated rat hepatocytes. BAY11-7082 plus BSO induced a greater p38 MAPK activation as compared to BAY11-7082 or BSO alone, and a p38 MAPK inhibitor protected against the synergistic toxicity. In summary, inhibition of NF-kappaB sensitizes liver cells to toxicity linked to GSH depletion, probably accelerating the processes of thiol homeostasis deregulation and induction of apoptosis through a mechanism mediated by p38 MAPK. PMID- 17920237 TI - Up-regulation of CYP1A/B in rat lung and liver, and human liver precision-cut slices by a series of polycyclic aromatic hydrocarbons; association with the Ah locus and importance of molecular size. AB - Exposure of precision-cut rat liver slices to six structurally diverse polycyclic aromatic hydrocarbons, namely benzo[a]pyrene, benzo[b]fluoranthene, dibenzo[a,h]anthracene, dibenzo[a,l]pyrene, fluoranthene and 1 methylphenanthrene, led to induction of ethoxyresorufin O-deethylase, CYP1A apoprotein and CYP1A1 mRNA levels, but to a markedly different extent. In liver slices, constitutive CYP1A1 mRNA levels were higher, as well as being markedly more inducible by PAHs, compared with CYP1B1, a similar profile to that observed in human liver slices following exposure to the PAHs. Increase in ethoxyresorufin O-deethylase and in CYP1A1 apoprotein levels was also observed when precision-cut rat lung slices were incubated with the same PAHs, the order of induction potency being similar to that observed in liver slices. Under the same conditions of exposure, CYP1B1 apoprotein levels were elevated in the lung. Up-regulation of CYP1A1 by the six PAHs correlated with their affinity for the Ah receptor, determined using the chemical-activated luciferase expression (CALUX) assay. It may be concluded that (a) precision-cut liver and lung slices may be used to assess the CYP1 induction potential of chemicals at the activity, apoprotein and mRNA levels; (b) rat is a promising surrogate animal for human in studies to evaluate CYP1 induction potential; (c) CYP1A1 is far more inducible than CYP1B1 in both rat liver and lung; (d) CYP1 up-regulation by PAHs is related to their affinity for the Ah receptor, and finally (e) computer analysis revealed that the ratio of molecular length/width is an important determinant of CYP1 induction potency among equiplanar PAHs. PMID- 17920236 TI - Ability of rabbit antiserum against crotapotin to neutralize the neurotoxic, myotoxic and phospholipase A2 activities of crotoxin from Crotalus durissus cascavella snake venom. AB - The toxicity of crotoxin, the major toxin of Crotalus durissus terrificus (South American rattlesnake) venom, is mediated by its basic phospholipase A(2) (PLA(2)) subunit. This PLA(2) is non-covalently associated with crotapotin, an acidic, enzymatically inactive subunit of the crotoxin complex. In this work, rabbit antiserum raised against crotapotin purified from Crotalus durissus cascavella venom was tested for its ability to neutralize the neurotoxicity of this venom and its crotoxin in vitro. The ability of this antiserum to inhibit the enzymatic activity of the crotoxin complex and PLA(2) alone was also assessed, and its potency in preventing myotoxicity was compared with that of antisera raised against crotoxin and PLA(2). Antiserum to crotapotin partially neutralized the neuromuscular blockade caused by venom and crotoxin in electrically stimulated mouse phrenic nerve-hemidiaphragm preparations and prevented the venom-induced myotoxicity, but did not inhibit the enzymatic activity of crotoxin and purified PLA(2). In contrast, previous findings showed that antisera against crotoxin and PLA(2) from C. d. cascavella effectively neutralized the neuromuscular blockade and PLA(2) activity of this venom and its crotoxin. The partial neutralization of crotoxin-mediated neurotoxicity by antiserum to crotapotin probably reduced the binding of crotoxin to its receptor following interaction of the antiserum with the crotapotin moiety of the complex. PMID- 17920238 TI - Expression levels of 63 p53-related genes add up to similar values in 24 different tissues and are unified in cancer. AB - The expression patterns of 62 genes interacting with p53 have been investigated in 24 normal and cancerous tissues using NIH's dbEST library. The expression levels of individual genes, such as the TTP53 gene itself, but also other genes, vary up to 33-fold among the 24 different tissues and no consistent pattern can be recognized. However, when expression levels for all 63 genes are summed, these "cumulated levels" are surprisingly constant over the 24 investigated normal tissues. In cancers, the variation is further reduced. Essentially, the cumulated expression levels in cancer are independent of those in normal tissue. We furthermore constructed a linear statistical classifier, i.e., a weighted sum of gene expression levels, which robustly distinguishes normal from cancer tissue independent of the particular kind of tissue. Thus, despite very large differences for individual genes and considerable changes during carcinogenesis, the cumulated expressions have narrowly defined levels. PMID- 17920239 TI - Can probability of genetic mutation be an indicator of clinical relevance? AB - NPM1 gene mutation evaluated on a population basis is a valuable and realistic tool to reflect the pathophysiological relevance of cancer. In a comparison of the NPM1 cDNA of human bladder cancer with its consensus sequence, we have found that a higher NPM1 sequence identity in a population is consistent with poor tumor differentiation, advanced tumor stage, and likelihood of recurrence. These data imply that "probability" of NPM1 mutation is an indicator of status of malignancy. PMID- 17920240 TI - Visual DNA -- identification of DNA sequence variations by bead trapping. AB - In this paper we describe a method that uses the nearly covalent strength biotin streptavidin interaction to attach a paramagnetic bead of micrometer size to a DNA molecule of nanometer size, scaling up the spatial size of a query DNA strand by a factor of 1000, making it visible to the human eye. The use of magnetic principles enables rapid binding and washing of detector beads, facilitating a readout of amplified DNA sequences in a few minutes. Here we exemplify the method on mitochondrial DNA variations using an array platform. Visual identification and documentation can be performed with an ordinary mobile phone equipped with a built-in camera. PMID- 17920241 TI - Brief and long maternal separations decrease corticosterone secretion in a lupus prone strain: dissociation from disease-related parameters. AB - Neonatal manipulations are known to alter the activity of the immune system and the hypothalamus-pituitary-adrenal (HPA) axis. This study was performed in order to examine whether brief and long maternal separations (BMS and LMS, respectively) interfere with the onset and development of murine lupus in NZB/NZWF1 females, and to determine whether the pattern of corticosterone (CORT) secretion throughout life is associated to the expression of the disease. Maternal separation was performed daily during postnatal days 1-14, lasting 15 min in the BMS group and 3h in the LMS group. Blood was sampled from the retro orbital plexus on the 9th week, and every other week, from 10th to 34th weeks of life, for detection of anti-nuclear antibodies (ANA) and anti-double-strand DNA (anti-dsDNA) antibodies, and for determination of CORT serum levels. Urine samples were collected on the 21st, 27th, 33rd and 37th weeks of life. There were no group differences in regard to disease-related parameters, but LMS females presented a tendency for late onset of anti-dsDNA antibodies. BMS and LMS mice exhibited reduced CORT levels compared to non-manipulated (NM) animals. There was a strong negative correlation between total mean CORT concentration and onset of ANA, and a strong positive correlation between total mean CORT concentration and life span only in the NM group. Neonatal manipulations appeared to eliminate these correlations; hence, both BMS and LMS modified basal CORT secretion and the association between glucocorticoids and immune activity in the NZB/NZWF1 mouse strain. PMID- 17920242 TI - Elucidating the consequences of chronic stress on immune regulation and behavior in rheumatoid arthritis. PMID- 17920244 TI - Evaluation of Alzheimer's disease by analysis of MR images using multilayer perceptrons and committee machines. AB - Alzheimer's disease is the most common cause of dementia, yet hard to diagnose precisely without invasive techniques, particularly at the onset of the disease. This work approaches image analysis and classification of synthetic multispectral images composed by diffusion-weighted magnetic resonance (MR) cerebral images for the evaluation of cerebrospinal fluid area and its correlation with the advance of Alzheimer's disease. The MR images were acquired from a unique volunteer with Alzheimer's, using an image system based on a clinical 1.5T tomographer. The classification methods are based on multilayer perceptrons and committee machines and the classification results are used to correlate clinical and imaging findings. The classification results are used to improve the usual analysis of the ADC map. PMID- 17920243 TI - Campylobacter jejuni infection increases anxiety-like behavior in the holeboard: possible anatomical substrates for viscerosensory modulation of exploratory behavior. AB - The presence of certain bacteria in the gastrointestinal tract influences behavior and brain function. For example, challenge with live Campylobacter jejuni (C. jejuni), a common food-born pathogen, reduces exploration of open arms of the plus maze, consistent with anxiety-like behavior, and activates brain regions associated with autonomic function, likely via a vagal pathway. As yet, however, little is known regarding the interface of immune sensory signals with brain substrates that mediate changes in behavioral states. To address this issue, we challenged mice with either C. jejuni or saline, and 7-8h later assessed anxiety-like behavior using the open holeboard, and used immunohistochemical detection of the protein c-Fos as an activation marker in the brain. C. jejuni treatment was associated with increased avoidance of the center regions of the holeboard, compared to saline-treated controls. Exposure to the holeboard induced activation in multiple brain regions previously implicated in anxiety-like behavior, including the lateral septum (LS), paraventricular (PVN) and dorsomedial hypothalamic nuclei (DMH), basolateral and central nuclei of the amygdala (BLA, CEA), bed nucleus of the stria terminalis (BST) and periaquiductal grey (PAG), compared to homecage controls. In C. jejuni-treated animals c-Fos induction also occurred in autonomic regions, as previously reported. The PVN, BLA, parts of the BST, medial prefrontal (mPFC) and anterior cingulate responded to both C. jejuni treatment and the holeboard, suggesting a role for these regions in the enhanced anxiety-like behavior observed. In saline-treated animals, anxiety-like behavior was predicted by activation in the CEA and BLA, whereas in C. jejuni-treated animals, c-Fos expression in the BST predicted the degree of anxiety-like behavior. These findings implicate the PVN, amygdala and BST as interfaces between gastrointestinal pathogenic challenge and brain regions that mediate behavioral responses to stress, and reinforce these nuclei as anatomical substrates by which viscerosensory stimuli can influence behavior. PMID- 17920246 TI - Continuous versus intermittent infusion of polymyxin B in the treatment of infections caused by multidrug-resistant Gram-negative bacteria. PMID- 17920245 TI - ITAREPS: information technology aided relapse prevention programme in schizophrenia. AB - ITAREPS presents a mobile phone-based telemedicine solution for weekly remote patient monitoring and disease management in schizophrenia and psychotic disorders in general. The programme provides health professionals with home telemonitoring via a PC-to-phone SMS platform that identifies prodromal symptoms of relapse, to enable early intervention and prevent unnecessary hospitalizations. Its web-based interface offers the authorized physician a longitudinal analysis of the dynamics and development of possible prodromes. This work presents preliminary findings from a one-year mirror-design follow-up evaluation of the programme's clinical effectiveness in 45 patients with psychotic illness. There was a statistically significant 60% decrease in the number of hospitalizations during the mean 283.3+/-111.9 days of participation in the ITAREPS, compared to the same time period before the ITAREPS entry (sign test, p<0.004). Variables significantly influencing the number of hospitalizations after the ITAREPS entry (medication compliance along with factors intrinsic to the ITAREPS, i.e. adherence to the programme and involvement of a family member) suggest a critical role of the programme in controlling the number of relapses and subsequent hospitalizations in psychosis. PMID- 17920247 TI - Synergistic effect between lycopene and ciprofloxacin on a chronic bacterial prostatitis rat model. AB - Traditionally, long-term antibiotic therapy has been the gold standard treatment for chronic bacterial prostatitis (CBP). However, the treatment outcome is not ideal and long-term administration of antibiotics can result in adverse effects and bacterial resistance. For these reasons, both patients and physicians are dissatisfied with the management of this disease and there is interest in phytotherapy and other alternative therapies. Lycopene, an extract of tomatoes, has been reported to have an anti-inflammatory effect via an antioxidative function. To evaluate the therapeutic effect of lycopene on CBP, we developed a CBP rat model treated with ciprofloxacin or lycopene, or both. After 2 weeks of treatment, results of microbiological cultures of the prostate and urine as well as histological findings of the prostate were analysed. The ciprofloxacin group and the lycopene/ciprofloxacin group showed a statistically significant decrease in bacterial growth and improvement in prostatic inflammation compared with the control group. The lycopene/ciprofloxacin group also showed a statistically significant decrease in bacterial growth and improvement in prostatic inflammation compared with the ciprofloxacin group. These results suggest that lycopene may have an additional (synergistic) effect with ciprofloxacin in the treatment of CBP. PMID- 17920248 TI - Alpha 1-noradrenergic system role in increased motivation for cocaine intake in rats with prolonged access. AB - In rodents, extended access to cocaine produces an escalation in cocaine self administration that has face and construct validity for human compulsive drug intake. Here we report that rats with six-hour access (long access, LgA) to cocaine self-administration produced a higher breakpoint for cocaine using a progressive-ratio schedule than rats with one-hour access (short access, ShA), and prazosin (alpha 1 receptor antagonist) reduced the higher breakpoint for cocaine in LgA rats. Additionally, the number of neurons with alpha 1-adrenergic receptor-like immunoreactivity in the bed nucleus of stria terminalis (BNST) was found to be much lower in LgA rats than in ShA and drug-naive rats. In contrast, UK14304 (alpha 2 receptor agonist) and betaxolol (beta 1 receptor antagonist) had no effect on cocaine self-administration in either group. The data suggest that activation of the alpha 1-noradrenergic system, perhaps in the BNST, is associated with increased motivation for cocaine in rats with extended access. PMID- 17920249 TI - Enhanced mixing in polyacrylamide gels containing embedded silica nanoparticles as internal electroosmotic pumps. AB - Many biosensors, including those based on sensing agents immobilized inside hydrogels, suffer from slow response dynamics due to mass transfer limitations. Here we present an internal pumping strategy to promote convective mixing inside crosslinked polymer gels. This is envisioned as a potential tool to enhance biosensor response dynamics. The method is based on electroosmotic flows driven by non-uniform, oscillating electric fields applied across a polyacrylamide gel that has been doped with charged colloidal silica inclusions. Evidence for enhanced mixing was obtained from florescence recovery after photobleaching (FRAP) measurements with fluorescein tracer dyes dissolved in the gel. Mixing rates in silica-laden gels under the action of the applied electric fields were more than an order of magnitude faster than either diffusion or electrophoretically driven mixing in gels that did not contain silica. The mixing enhancement was due in comparable parts to the electroosmotic pumping and to the increase in gel swelling caused by the presence of the silica inclusions. The latter had the effect of increasing tracer mobility in the silica-laden gels. PMID- 17920250 TI - Effect of chain density and conformation on protein adsorption at PEG-grafted polyurethane surfaces. AB - Polyurethanes were modified using monobenzyloxy polyethylene glycol (BPEG) which possesses a bulky hydrophobic benzyloxy group at one end and a hydroxyl group at the other end as a preconstructed BPEG layer, and poly(ethylene glycol) (PEG) and monomethoxyl poly(ethylene glycol) (MPEG) with various chain lengths as fillers. Our objective was to investigate the effect of PEG graft density and conformation on protein adsorption at PEGlated surface. The graft density was estimated by a chemical titration method. The combination of ATR-FTIR, AFM and titration results provide evidences that the graft density can be increased by backfilling PEG or MPEG to a BPEG layer. However, fibrinogen and albumin adsorption significantly increased on all surfaces after PEG or MPEG backfilling. We conclude that the conformation of hydrophobic benzyloxy end groups of the BPEG layer plays a key role. The benzyloxy end groups of preconstructed PEG chains stretch to the surface after PEG backfilling, which possibly accounts for the observed increase in protein adsorption. The BPEG conformation change after backfilling with PEG or MPEG was also suggested by contact angles. Additionally, protein adsorption was slightly influenced by the length of filler, suggesting a change in surface morphology. PMID- 17920251 TI - [Focal nodular hyperplasia involving portal cavernoma]. AB - Focal nodular hyperplasia is a rare hepatic tumor in children representing only 2% of all pediatric hepatic tumors. We report a case in teen-ager who had a symptomatic portal cavernoma at 9 years of age, and had been treated by porto mesenteric surgical shunt. At 16 years, liver ultrasounds revealed multiple hepatic nodules. Focal nodular hyperplasia was suggested by CT scan and magnetic resonance imaging. Given the symptoms and the unusual and multiple lesions, diagnosis was confirmed by surgical biopsy. FNH pathogeny is discussed in this context. PMID- 17920253 TI - [Errors for delivery medications through gastrostomy devices: study of 109 children]. AB - Gastrostomy is frequently used in clinical practice for drug administration. However modalities of drug administration via a gastrostomy device have been poorly studied and remain uncodified. AIM: To assess the mode of administration of drugs as well as errors associated with the use of a gastrostomy devices for drug delivery in children. PATIENTS AND METHODS: Mode of administration of drugs was studied in 109 children (mean age 8.4+/-5.5 years, 72% neurologically impaired, 41% institutionalized children). A questionnaire was filled in by parents and/or caregivers. Errors of administration were classified as follows: galenic, due to preparation, physico-chemical interaction, lack of flushing the tube. Factors influencing the occurrence of errors were studied: living at home or in an institution, underlying disease, number of drugs administered via gastrostomy device. RESULTS: Errors were frequently observed: galenic (47%), due to preparation (42%), physico-chemical interaction (51%), lack of flushing (10%). Errors occurred more frequently in institutionalized children compared to children living at home (78 versus 25%, P<0.0005). Galenic errors were more frequent in neurologically impaired children (57 versus 30%, P<0.005). CONCLUSION: Many medications are administrated via gastrostomy tube in children. Errors are frequently observed and potentially dangerous. PMID- 17920252 TI - [Pneumococcal infection among community-acquired pneumonia. A retrospective study of 230 hospitalized children]. AB - OBJECTIVES: To find arguments in favour of pneumococcal origin in community acquired pneumonia. POPULATION AND METHODS: A retrospective analysis of the files of 230 children hospitalized between January 1st 1999 and June 30th 2001 for community acquired pneumonia was performed. The files were classified into 3 subgroups: I (N=7), confirmed (positive blood culture); II (N=134), probable (biological arguments); III (N=89), possible pneumococcal infection. Age of the children was also taken into consideration. RESULTS: All children in the subgroup I had fever>39 degrees C at admission and at least 1 of the 3 criteria (WBC> or=20.10(9)/l, neutrophils > or =10.10(9)/l, C-reactive protein level> or =60 mg/l). Dyspnea was more frequently asthmatiform in the subgroup III. Chest X-ray was not contributive. Before admission, 39% of the children were given one or several antibiotics, and so some of patients belonging to the subgroups II and III could have been infected by pneumococcus without possibility to confirm that. CONCLUSION: Results of this analysis suggest that some criteria may be useful for selecting initial antibiotherapy even though systematic early specific antipneumococcal immunization should reduce the frequency of this infection. PMID- 17920254 TI - Heterogeneous effects of gene polymorphism on type 2 diabetes risk: lesson from the PPAR gamma 2 Pro12Ala. PMID- 17920255 TI - Effect of Ninjin-yoei-to (Rensheng-Yangrong-Tang) on olfactory behavior after olfactory nerve transection. AB - Ninjin-yoei-to (NYT), a Japanese traditional medicine, is used to treat athrepsia due to surgery, anorexia, cold constitution, and anemia. There are reports of the effects of NYT on the nervous system; however, there have been no behavioral studies of the effect of NYT on olfactory function. The olfactory system undergoes continuous replacement of sensory neurons. Morphologic and behavioral studies have shown that the olfactory system recovers after bilateral olfactory nerve transection (BNX). However, in the humans, olfactory function does not always recover. In this study, we examined the effect of oral NYT on behavioral recovery after BNX. Fourteen mice were subjected to BNX. The regular diet was mixed with 2% NYT (NYT diet). Mice were separated into two groups; seven mice were fed the regular diet (control group), and seven mice were fed the NYT diet (NYT group). NYT was administered beginning 7 days prior to BNX and continuing for 35 days after BNX. Mice in both groups had free access to food and water. Olfactory function was evaluated by testing each mouse's ability to avoid cotton balls treated with acetic acid. After BNX, mice lost their ability to avoid cotton balls treated with acetic acid. In the control group, the time for behavioral recovery after BNX was 28 days. In the NYT group, the time for behavioral recovery after BNX was 21 days. NYT hastened behavioral recovery after BNX. NYT may have therapeutic benefits for patients with olfactory disorders. PMID- 17920256 TI - Improvements with tiotropium in COPD patients with concomitant asthma. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) and asthma have different diagnostic criteria and treatment paradigms. Both are common and can occur in the same patient. We sought to determine the spirometric effects of tiotropium in COPD patients with concomitant asthma. METHODS: A 12-week randomized, double-blind, placebo-controlled, parallel group trial with tiotropium 18 mcg daily was performed. Patients continued usual respiratory medications except for inhaled anticholinergics. INCLUSION CRITERIA: Physician diagnosis of COPD and asthma, age >or= 40 years, smoking >10 pack years, post bronchodilator forced expiratory volume in 1s (FEV(1))<80% predicted, FEV(1)/forced vital capacity (FVC)<70%, >or= 12%, and >or= 200 ml increase in FEV(1) following inhaled bronchodilator, treatment with inhaled steroids >or= 1 year. Spirometry was measured serially for 6h on days 1, 29 and 85. RESULTS: Four hundred and seventy-two patients were randomized. Baseline characteristics were balanced. Mean age=59.6 years, 61.4% were men, and FEV(1)=1.55l (53.0% predicted). Improvements at 12 weeks with tiotropium were observed for the primary endpoint FEV(1) area under the curve (AUC) from 0 to 6h (difference=186+/ 24 ml, p<0.001) and for morning pre-dose FEV(1) (difference=98+/-23 ml, p<0.001). Significant differences in favor of tiotropium were observed for pre-dose FVC (difference=128+/-34 ml, p<0.001) and FVC AUC 0-6h (difference=232+/-35 ml, p<0.001). Compared to baseline, the mean weekly number of daily puffs of prn salbutamol was reduced by 0.05+/-0.12 puffs/day in the placebo group and by 0.50+/-0.12 puffs/day in the tiotropium group at week 12 (p<0.05). CONCLUSIONS: Patients with COPD and concomitant asthma achieve spirometric improvements with tiotropium along with symptomatic benefit as seen by reduced need for rescue medication. PMID- 17920257 TI - Effectiveness of omalizumab (Xolair) in the first patients treated in real-life practice in France. AB - OBJECTIVE: To describe omalizumab (Xolair) effectiveness in the first patients treated on compassionate grounds before its commercialisation in France. METHODS: In a historic-prospective study, data were obtained by questionnaire from the physicians whose patients had received a nominative temporary use authorisation (ATU) for omalizumab from July 2003 to January 2006. Anonymised patient data regarding demographics, asthma-related treatments and events in the year previous to the start of omalizumab treatment as well as the details of omalizumab treatment itself were obtained at inclusion. Follow-up data at more than 3 months following inclusion were also obtained and regarded asthma-related treatment (including omalizumab), events and undesirable effects suspected to be linked to omalizumab treatment. RESULTS: Data were obtained for 147 of the 154 patients treated via ATU. 31.3% received inappropriate monthly doses of omalizumab. Of the 28 patients (19%) who discontinued for unsatisfactory therapeutic effect, 7 were treated for less than the 16 weeks recommended to evaluate efficacy and 9 who were treated for a longer period of time were underdosed. During the treatment period and compared to the previous year, patients with follow-up data at 5 months or more had experienced 62% fewer exacerbations requiring oral corticosteroids, 65% fewer emergency department visits and 29% fewer hospitalisations per year. The nature of adverse effects reported was similar to that reported in omalizumab clinical trials. CONCLUSION: Results strongly suggest that omalizumab in the first patients treated in real-life setting provided a similar benefit to that observed in clinical trials. Underdosing of patients may limit this therapeutic effect. PMID- 17920258 TI - Australian heroin seizures and the causes of the 2001 heroin shortage. AB - This paper uses Australian heroin seizure data, along with estimates of the size of the Australian heroin market to evaluate the impact of drug law enforcement on the 2001 Australian heroin shortage from the percentage of the market seized. It also critically examines international heroin production trends and published reports on the causes of the Australian heroin shortage. Its conclusion is that previous studies may have overstated the success of drug law enforcement and that the most likely explanation for Australia's 2001 heroin shortage was a significant decline in heroin production world-wide, due to a general move away from heroin production in the countries of Southeast Asia and the prohibition on opium growing by the Taliban regime in Afghanistan. PMID- 17920259 TI - Functional organisation of the genome during interphase. AB - Mammalian genomes are partitioned in chromosomes and contain millions of base pairs that encode for tens of thousands of protein-coding genes ( approximately 30,000 in humans) and many more non-coding transcripts. Gene forests interspersed with gene deserts in the linear DNA sequence may segregate into domains that are more or less permissive for transcription within the three-dimensional nucleus. Functional attachments to various nuclear landmarks are thought to organise the architectural folding of the chromosome fibre. The position of a gene within the nucleus can favour its silencing or activation and the efficiency with which its products are processed or transported to the cytoplasm. The stochastic properties of genome organisation may contribute to cell-type-specific gene expression and to the dynamic responses that occur during differentiation and adaptation to the environment. The first steps for high-throughput mapping of chromosome architecture in specific cell types are under way to help understand the mechanisms by which genome architecture may regulate gene expression. PMID- 17920260 TI - MAD contortions: conformational dimerization boosts spindle checkpoint signaling. AB - Almost two decades after their identification, the components of the mitotic checkpoint are finally revealing their structural secrets. The activation of Mad2, a central piece of the checkpoint protein machinery, is linked to the rare ability of this protein to adopt two distinct topologies. Current models of checkpoint function propose that the topological transition between the two states of Mad2 is rate limiting for checkpoint activation and is accelerated through a self-activation process based on the direct interaction of the two Mad2 conformers. These models add a molecular framework to an old theory that depicts kinetochores as catalysts in the generation of the mitotic checkpoint signal. PMID- 17920261 TI - HER-3 in colorectal tumourigenesis: from mRNA levels through protein status to clinicopathologic relationships. AB - INTRODUCTION: Colorectal cancer is a major cause of cancer mortality in the Western world. Although HER-3 signalling is known to be implicated in colorectal carcinogenesis, the significance of its expression, localisation and phosphorylation remains elusive. METHODS: Quantitative RT-PCR for HER-3 mRNA and immunohistochemistry for HER-3 and phosphorylated HER-3 (pHER-3) protein were performed in normal tissue, adenomas and carcinomas from 140 patients with colorectal cancer. RESULTS: HER-3 was detected both in the cytoplasm and nucleus, whereas pHER-3 was observed in the nucleus and membrane of cells. A possible switch in HER-3 topography from the nucleus to the cytoplasm during colorectal tumourigenesis is suggested. The expression of pHER-3 did not differ significantly in normal tissue, adenomas and carcinomas, but was related to disease stage. HER-3 mRNA overexpression was significantly associated with decreased time to disease progression. It was also correlated with higher median age, left colon and rectal tumour sites and lymph node involvement. CONCLUSION: We postulate that HER-3 is critically involved in colorectal tumourigenesis and its expression/phosphorylation might be of prognostic significance. PMID- 17920262 TI - Reclamation of chitinous materials by bromelain for the preparation of antitumor and antifungal materials. AB - The main purpose of this research was to investigate the antitumor and antimicrobial activities of the chitooligosaccharides containing hydrolyzates obtained from the hydrolysis of chitinous materials (such as chitin, chitosan, and squid pen) by bromelain. The optimum preparations were gained in the hydrolysates of squid pen powder hydrolyzed at pH 5, 37 degrees C for 2 days by 0.1% bromelain. The hydrolysates had an 80% inhibitory activity on phyto pathogenic mold Fusarium oxysporum. Chitooligosaccharides were recovered from the hydrolysates and were used for tumor cell surviving test. Surviving rate of the human leukemic U937 cells was reduced to 69% by the chitooligosaccharides. The solution of 0.1% of water-soluble chitosan was also hydrolyzed for 1 day at pH 5, 37 degrees C by bromelain. The resultant hydrolysates contained the highest chitooligosaccharides, which had inhibitory effect on Bacillus subtilis and also had 40% inhibitory activity on human pathogenic mold Aspergillus fumigatus. Surviving rate of mouse CT26 colorectal adenocarcinoma cells was reduced to 57% by the chitooligosaccharides. This is the first publication of enzymatic reclamation of squid pen (fishery processing waste) for the preparation of antitumor and antimicrobial materials. PMID- 17920263 TI - Determination of a relationship between chitinase activity and microbial diversity in chitin amended compost. AB - By using denaturing gradient gel electrophoresis (DGGE) and simultaneously measuring the enzymatic activity of chitinase, we could link genetic diversity of the indigenous microbial communities with chitinase activity in compost samples. A garden/park waste compost and a source separated organic household waste compost, showed different genetic diversity as measured by PCR-DGGE of total DNA extracted from the composts. The household waste compost had the highest chitinase activity. To increase chitinase activity, the two composts were amended with chitin. This addition induced a change in both the bacterial and fungal genetic diversity when compared to the non-amended compost samples. Likewise, both composts reacted to the addition of chitin with an increase in chitinase activity. Thus, a relationship between genetic diversity and chitinase activity was established for the composts in question. The N-mineralization in the household waste compost was apparently increased by the addition of chitin, while such an effect was not observed in the garden/park waste compost. PMID- 17920264 TI - A novel raw starch digesting alpha-amylase from a newly isolated Bacillus sp. YX 1: purification and characterization. AB - This study reports the purification and characterization of a novel raw starch digesting alpha-amylase from a newly isolated Bacillus sp. YX-1. Maximum alpha amylase activity (53 U mL(-1)) was obtained at 45 degrees C after 44 h of incubation. The enzyme was purified using ammonium sulfate precipitation, ion exchange and gel filtration chromatography, and showed a molecular weight of 56 kDa by SDS-PAGE. This enzyme exhibited maximum activity at pH 5.0, performed stability over a broad range of pH 4.5-11.0, and was optimally active at 40-50 degrees C. The enzyme preparation had a strong digesting ability towards various raw starches and efficiently hydrolyzed raw corn starch at a concentration of 20% and pH 5.0, which were normally used in the starch industries, in a period of 12h. By analyzing its partial amino acid sequences, the enzyme was proposed to be a novel alpha-amylase. PMID- 17920265 TI - Effect of reactive substrates used for the removal of phosphorus from wastewater on the fertility of acid soils. AB - Reactive substrates used in filter systems can reduce phosphorus (P) pollution and, once saturated with P, may be recycled in agriculture. These substrates are usually calcium carbonate derivates with high pH values, which may be particularly beneficial for acid soils. Three reactive substrates (Filtra P, Polonite and wollastonite) saturated with P were used as amendments to an acid soil in a pot experiment. Substrate amendments tended to improve ryegrass yield and P uptake compared with control and potassium phosphate treatments. Polonite produced the highest yield/amendment ratio, while Polonite and Filtra P significantly increased the concentrations of P and Ca in the ryegrass. Addition of all three substrates increased the pH, AL-extractable P and cation exchange capacity of soils during the experiment. These substrates can therefore be applied to acid soils in order to recycle P and improve soil properties. PMID- 17920266 TI - Structure-activity relationship study on the 6-membered heteroaromatic ring system of diphenylpyrazine-type prostacyclin receptor agonists. AB - A series of prostacyclin receptor agonists was prepared by modifying the central heteroaromatic ring of lead compound 2, and a docking study was performed to investigate their structure-activity relationships by using a homology-modeled structure of the prostacyclin receptor. Compound 2 and its derivatives could be docked to the prostacyclin receptor in two ways depending on the position of the nitrogen atom within the heteroaromatic ring. Furthermore, hydrogen bonding between the nitrogen atom in the heteroaromatic ring and the hydroxyl group of Ser20 or Tyr75 of the receptor appears to be important for the potent expression of biological activity. PMID- 17920267 TI - A simple method for the preparation of PEG-6-mercaptopurine for oral administration. AB - A new and efficient method for the synthesis of PEG-6-mercaptopurine is described. The key feature of the proposed approach is the protection of the thiol group against metabolic inactivation. Preliminary in vivo and in vitro evaluations of the macromolecular prodrug have been carried out. PMID- 17920268 TI - Rational design of N-alkyl derivatives of 2-amino-2-deoxy-d-glucitol-6P as antifungal agents. AB - N-Alkyl and N,N-dialkyl derivatives of 2-amino-2-deoxy-d-glucitol-6P (ADGP) were synthesized and found to inhibit growth of human pathogenic fungi (MICs in the 0.08-0.625mgmL(-1) range for the most active compounds). It was thus shown that N alkylation of ADGP provides novel inhibitors of a fungal enzyme, glucosamine-6P synthase, exhibiting higher antifungal activity than the parent compound, due to the increased lipophilicity and better uptake by fungal cells. PMID- 17920270 TI - The experiences of patients and relatives/significant others of overcrowding in accident and emergency in Ireland: a qualitative descriptive study. AB - Overcrowding in Accident and Emergency Departments (A&E) in Ireland has reached crisis proportions since the dawning of the new millennium. Although this phenomenon is not unique to Ireland and many authors have suggested causes and management strategies to deal with these crises, little appears to have been written regarding the experiences of patients or their families waiting in the A&E. The aim of this qualitative descriptive study was to describe the experiences of patients and/or their relatives/significant others who had spent 12 h or more in A&E awaiting admission to hospital. Four patients and three relatives/significant others volunteered to participate in the study. Participants described the A&E departments as resembling a disaster zone or a hospital scene from a third world country. Descriptions portrayed an environment that was overcrowded, dirty and lacking in resources. Participants were generally positive in their attitudes towards the care they received, but some descriptions appeared to suggest that the quality of care was not always ideal. Recommendations from participants included reduced waiting times with a maximum of 6 h from admission to transfer or discharge; better communications systems with perhaps a liaison person who could advise them about the expected duration of stay in A&E and what was happening regarding their care; and better privacy and security within the departments. PMID- 17920269 TI - Trends in childhood injury mortality in a developing country: United Arab Emirates. AB - OBJECTIVE: To describe the epidemiology of a leading cause of childhood mortality in Al-Ain, United Arab Emirates (UAE)--injury. To examine trends across types of injury, as well as the mechanisms of injury leading to death, by age groups, gender, citizenship, and explore mortality rates and make global comparisons. DESIGN: This is a retrospective, descriptive, statistical analysis of unlinked hospital data. SETTING: Al-Ain and Tawam Hospitals, and Preventive Medicine Department, Al-Ain, United Arab Emirates. SUBJECTS: All cases that met the conditions established for the study: fell within the age group of 0 to 14 years, suffered from injuries, and were admitted to either Al-Ain or Tawam hospitals and subsequently died within the studied time period of 1 January 1995 to 31 December 2004. RESULTS: A total of 7204 deaths were reported in children below 15 years during the studied time period. Of these cases, 2150 children died due to injury, comprising 29.8% of total deaths. Further analysis showed that road traffic injuries were the most frequent cause of injury leading to death (68.3%). Overall injury death rates were higher in non-citizens (54.5%) than in citizens (45.5%); and males had a higher incidence, specifically a 2.1:1 ratio, than females. Children 5 to 14 years had the highest frequency of injury deaths. Overall, injury mortality rates exhibited a decreasing, though fluctuating, trend during the studied period at a rate that is comparable to those in other developed nations such as New Zealand and USA. CONCLUSION: The present study reveals that the burden of injury deaths among children below 15 years is significant; and injuries exist in every form and affect every age group, and gender. The high burden of injuries on children in the UAE demands the attention of the health community, including policy makers. An understanding of the trends such as those presented in this study, for instance that injuries from road traffic are prominent, will assist in the development of interventions to address this growing concern. Furthermore, similarities in rates of UAE with other developed countries signify the potential for appropriate responses to lower the burden of injuries on children in the future. PMID- 17920271 TI - Strategies used to walk through a moving aperture. AB - The objectives of the study were to determine what strategy (pursuit or interception) individuals used to pass through an oscillating target and to determine if individuals walked towards where they were looking. Kinematic and gaze behaviour data was collected from seven healthy female participants as they started at one of five different starting positions and walked 7 m towards an oscillating target. The target was a two-dimensional 70 cm aperture made by two 76 cm wide doors and oscillated between two end posts that were 300 cm apart. In order to quantify the objectives, target-heading angles [Fajen BR, Warren WH. Behavioral dynamics of steering, obstacle avoidance, and route selection. J Exp Psychol Hum Percept Perform 2003;29(2):343-62; Fajen BR, Warren WH. Visual guidance of intercepting a moving target on foot. Perception 2004;33:689-715] were calculated. Results showed that the participants used neither an interception nor a pursuit strategy to successfully pass through the moving aperture. The participants steered towards the middle of the pathway prior to passing through the middle of the aperture. A cross correlation between the horizontal gaze locations and the medial/lateral (M/L) location of the participants' center of mass (COM) was performed. The results from the cross correlation show that during the final 2s prior to crossing the aperture, the participants walked where they were looking. The findings from this study suggest that individuals simplify a task by decreasing the perceptual load until the final stages. In this way the final stages of this task were visually driven. PMID- 17920272 TI - The effect of trunk flexion on able-bodied gait. AB - This study examined the effect of sagittal trunk posture on the gait of able bodied subjects. Understanding the effect of trunk posture on gait is of clinical interest since alterations in trunk posture often occur with age or in the presence of spinal pathologies, such as lumbar flatback. Gait analysis was conducted on 14 adults walking at self-selected slow, normal, and fast walking speeds while maintaining three trunk postures: upright, and with 25+/-7 degrees and 50+/-7 degrees of trunk flexion from the vertical. During trunk-flexed gait, subjects adopted a crouch posture characterized by sustained knee flexion during stance and an increase in ankle dorsiflexion and hip flexion angles. During stance, these kinematic adaptations produced a posterior shift in the positions of the trunk and pelvis, which helped to offset the anterior shift in the trunk mass that occurred with trunk flexion. In this way, kinematic adaptations may have been used to maintain balance by shifting the body's center of mass to a position similar to that of upright walking. These changes in lower limb joint kinematics created a phase lag in the position of the hip joint center relative to that of the ankle joint center in the sagittal plane. Alterations in the sagittal alignment of the hip and ankle joint positions were associated with a phase lag in the vertical position, velocity, and acceleration of the body's center of mass (BCOM) relative to upright walking. Since the vertical ground reaction force (GRF(v)) is proportional to the vertical acceleration of the BCOM, significant changes were also seen in the GRF(v) during trunk-flexed gait. In summary, kinematic adaptations necessary to maintain dynamic balance altered the trajectory and acceleration of the BCOM in the vertical direction, which was reflected in the GRF(v). The results of this study may help clinicians better understand the nature and impact of compensatory mechanisms in patients who exhibit trunk-flexed postures during gait. PMID- 17920273 TI - The benefits of using diverse animal models for studying pertussis. AB - Pertussis, a respiratory disease caused by infection with Bordetella pertussis, represents one of the most devastating diseases in infants and young children worldwide. Significant research efforts over the last five decades have led to the introduction of two types of vaccines, which are now available worldwide and which have significantly reduced the global incidence of pertussis. The use of animal models and, in particular, the mouse model has benefited in the development of these vaccines tremendously. However, open questions regarding the duration of immunity, the type of immune response needed for protection and the role of mucosal and innate immunity in disease protection still remain. Here, we review the various animal models available currently and their benefits for studying this important disease. PMID- 17920274 TI - Campylobacter flagella: not just for motility. AB - Campylobacter jejuni and Campylobacter coli are among the major causes of diarrheal disease worldwide. The motility imparted by the polar flagella of these pathogens is required for colonization of the mucus lining of the gastrointestinal tract. However, recent studies have revealed a more complex role for flagella in Campylobacter pathogenesis that includes the ability to secrete non-flagellar proteins that modulate virulence and the co-regulation of secreted and non-secreted virulence factors with the flagella regulon. Campylobacter flagellins are heavily glycosylated and changes in glycan composition affect autoagglutination and microcolony formation on intestinal epithelial cells; these traits are associated with disease in an animal model. Here, these recent advances in our understanding of the multifaceted role of flagella in Campylobacter virulence are summarized. PMID- 17920275 TI - The elusive activity of the Yersinia protein kinase A kinase domain is revealed. AB - Yersinia spp. pathogens use their type III secretion system to translocate effectors that manipulate host signaling pathways during infection. Although molecular targets for five of the six known Yersinia effectors are known, the target for the serine/threonine kinase domain of Yersinia protein kinase A (YpkA) has remained elusive. Recently, Navarro et al. (2007) demonstrated that YpkA phosphorylates Galphaq, and inhibits Galphaq-mediated signaling. Inhibition by YpkA could contribute to one of the most documented symptoms of Yersinia pestis infection, extensive bleeding. PMID- 17920276 TI - Microtiter plates as mini-bioreactors: miniaturization of fermentation methods. AB - In the past decade, the use of microtiter plates for microbial growth has become widespread, particularly in industry. In parallel, research in academia has provided a thorough insight into the complex relation between well dimensions, culture volumes, orbital shaking conditions and surface tension on the one hand, and oxygen-transfer rates and degrees of mixing on the other. In this review, I will discuss these fundamental issues and describe the current applications of microtiter plates in microbiology. Microtiter plates can now be considered a mature alternative to Erlenmeyer shake flasks. PMID- 17920277 TI - ABC transporters: how small machines do a big job. AB - Transporters from the ATP-binding cassette (ABC) superfamily operate in all organisms, from bacteria to humans, to pump substances across biological membranes. Recent high-resolution views of ABC transporters in different conformational states provide clues as to how ATP might be used to drive the structural reorganizations that accompany membrane transport. Importantly, it now appears that a putative translocation pathway running through the center of the transporter might be gated alternately, either at the inside or the outside of the cytoplasmic membrane, coupling substrate translocation to a cycle of ATP dependent conformational changes. ATP binding and ATP hydrolysis have distinct roles in this cycle: binding favors the outward-facing orientation, whereas hydrolysis returns the transporter to an inward-facing conformation. PMID- 17920278 TI - Bare rudiments of notch signaling: how receptor levels are regulated. AB - The Notch signaling pathway, a prototype for high-fidelity intercellular signaling mechanisms, regulates cell fate and other fundamental processes. Notch receptors are activated by transmembrane ligands present on neighboring cells through ligand-dependent proteolysis. This liberates the receptor intracellular domain, creating a transcriptional coactivator. Although tremendous progress towards elucidating the Notch activation mechanism and Notch-dependent processes has been reported, perhaps the most fundamental issue of Notch signaling is still poorly understood -- how cell-type-specific Notch-receptor expression patterns are established and regulated. This is especially true for the four mammalian Notch receptors (Notch1-4), which exhibit unique and overlapping expression patterns. However, recent studies have demonstrated the transcriptional control of Notch receptor levels as an essential determinant of Notch signaling. PMID- 17920279 TI - The PI3K inhibitor arsenal: choose your weapon! AB - Owing to its widespread activation in inflammation and cancer, a growing appreciation of the therapeutic potential of inhibitors of the phosphoinositide 3 kinase (PI3K) pathway has stimulated intense interest in compounds with suitable pharmacological profiles. These are primarily directed toward PI3K itself. However, as class I PI3Ks are also essential for a range of normal physiological processes, broad spectrum PI3K inhibition could be poorly tolerated. In recent years, patents describing a new generation of PI3K inhibitors have started to appear, with a particular focus on the development of compounds with enhanced isoform selectivity for use as anti-cancer and anti-inflammatory therapies. However, challenges remain for the efforts to pharmacologically target this enzyme family in a successful manner. PMID- 17920280 TI - That which does not kill me makes me stronger: adapting to chronic ER stress. AB - Cells respond to the accumulation of unfolded proteins by activating signal transduction cascades that improve protein folding. One example of such a cascade is the unfolded protein response (UPR), which senses protein folding stress in the endoplasmic reticulum (ER) and leads to improvement in the protein folding and processing capacity of the organelle. A central paradox of the UPR, and indeed of all such stress pathways, is that the response is designed to facilitate both adaptation to stress and apoptosis, depending upon the nature and severity of the stressor. Understanding how the UPR can allow for adaptation, instead of apoptosis, is of tremendous physiological importance. Recent advances have improved our understanding of ER stress and the vertebrate UPR, which suggest possible mechanisms by which cells adapt to chronic stress. PMID- 17920281 TI - The use of ligand-based de novo design for scaffold hopping and sidechain optimization: two case studies. AB - This paper describes the application of de novo design utilizing exclusively ligand information. In the current approach, ligand design criteria, including pharmacophores, similarity and desired properties are applied as part of a fitness function driving the design process, instead of using them as filters after the process. This allows relevant parts of chemical space to be explored more efficiently. Two case studies of successful ligand design are also presented, one aimed at scaffold hopping, the other for exploring substitution patterns around a novel scaffold. PMID- 17920282 TI - Discovery of novel alpha-glucosidase inhibitors based on the virtual screening with the homology-modeled protein structure. AB - Discovery of alpha-glucosidase inhibitors has been actively pursued with the aim to develop therapeutics for the treatment of diabetes and the other carbohydrate mediated diseases. We have been able to identify 13 novel alpha-glucosidase inhibitors by means of a computer-aided drug design protocol involving homology modeling of the target protein and the virtual screening with docking simulations under consideration of the effects of ligand solvation in the binding free energy function. Because the newly discovered inhibitors are structurally diverse and reveal a significant potency with IC(50) values lower than 50 microM, all of them can be considered for further development by structure-activity relationship studies or de novo design methods. Structural features relevant to the interactions of the newly identified inhibitors with the active site residues of alpha-glucosidase are discussed in detail. PMID- 17920283 TI - Increased metabolism in the R6/2 mouse model of Huntington's disease. AB - Huntington's disease (HD) is a hereditary disorder characterized by personality changes, chorea, dementia and weight loss. The cause of this weight loss is unknown. The aim of this study was to examine body weight changes and weight regulating factors in HD using the R6/2 mouse model as a tool. We found that R6/2 mice started losing weight at 9 weeks of age. Total locomotor activity was unaltered and caloric intake was not decreased until 11 weeks of age, which led us to hypothesize that increased metabolism might underlie the weight loss. Indeed, oxygen consumption in R6/2 mice was elevated from 6 weeks of age, indicative of an increased metabolism. Several organ systems that regulate weight and metabolism, including the hypothalamus, the stomach and adipose tissue displayed abnormalities in R6/2 mice. Together, these data demonstrate that weight loss in R6/2 mice is associated with increased metabolism and changes in several weight-regulating factors. PMID- 17920284 TI - Intrastriatal inhibition of aromatic amino acid decarboxylase prevents l-DOPA induced dyskinesia: a bilateral reverse in vivo microdialysis study in 6 hydroxydopamine lesioned rats. AB - l-3,4-dihydroxyphenylalanine (l-DOPA)-induced dyskinesia consists of involuntary choreiform and dystonic movements. Here we report whether intrastriatal l-DOPA itself is able to trigger dyskinetic behavior and which role the neurotransmitter dopamine (DA) and its metabolites play. Intrastriatal l-DOPA as well as DA administration at the 6-hydroxydopamine (6-OHDA) lesioned side led to a significant appearance of dyskinetic behavior, whereas DA metabolites were ineffective. Intrastriatal inhibition of the enzyme aromatic amino acid decarboxylase (AADC) by benserazide prevented the appearance of l-DOPA-induced dyskinetic movements at the lesioned side. Principle component analysis of DA and DA metabolite levels with dyskinesia scores after l-DOPA/benserazide (6/15 mg/kg) administration indicated a significant correlation only for DA, whereas DA metabolites did not show any significant correlation with the occurrence of dyskinetic behavior. We conclude that intrastriatal l-DOPA itself is not able to induce dyskinetic movements, whereas the increase of intrastriatal DA levels is instrumental for l-DOPA- and DA-induced dyskinetic behavior. PMID- 17920286 TI - Incidence, risk factors, and prognosis of postoperative hyperbilirubinemia after heart transplantation. AB - OBJECTIVE: The clinical significance of postoperative hyperbilirubinemia after heart transplantation has not been reported. Here, we sought to evaluate the incidence, risk factors, and prognosis of postoperative hyperbilirubinemia after heart transplantation. METHODS: Between 1987 and 2005, 256 consecutive patients undergoing heart transplantation were studied prospectively. Postoperative hyperbilirubinemia was defined as occurrence of a serum total bilirubin concentration of more than 3mg/dl in any measurement during the postoperative period. Logistic regression was done to identify possible risk factors for postoperative hyperbilirubinemia and hospital mortality. RESULTS: Overall incidence of postoperative hyperbilirubinemia was 57%. Among all patients, there were 35 hospital deaths (14%). In patients with postoperative hyperbilirubinemia, the mean onset time was 2.4+/-4.4 days after transplantation and the mean peak serum total bilirubin was 10.1+/-10.4 mg/dl. Development of postoperative hyperbilirubinemia was associated with a higher mortality (21% vs 5%, P<0.001 by Fisher's exact test). The onset time of postoperative hyperbilirubinemia, the peak serum total bilirubin level, and the time at which the peak bilirubin level was reached were associated with hospital mortality. Old donor age, valvular heart disease, high right atrial pressure, use of mechanical ventilation before transplant, and ascites at transplant were the significant risk factors for postoperative hyperbilirubinemia. CONCLUSIONS: Postoperative hyperbilirubinemia is common in patients undergoing heart transplantation and is associated with high hospital mortality. Patients with valvular heart disease, high preoperative right atrial pressure, and ascites at transplant, who then receive an old donor heart, are at greater risk for development of postoperative hyperbilirubinemia. PMID- 17920285 TI - Oxidatively modified RNA in mild cognitive impairment. AB - Studies show increased oxidative damage in the brains of subjects with Alzheimer's disease (AD) and mild cognitive impairment (MCI). To determine if RNA oxidation occurs in MCI, sections of hippocampus/parahippocampal gyrus (HPG) from 5 MCI, 5 late stage AD (LAD) and 5 age-matched normal control (NC) subjects were subjected to immunohistochemistry using antibodies against 8-hydroxyguanine (8 OHG) and 1-N2-propanodeoxyguanosine (NPrG). Confocal microscopy showed 8-OHG and NPrG immunostaining was significantly (p<0.05) elevated in MCI and LAD HPG compared with NC subjects and was predominately associated with neurons identified using the MC-1 antibody that recognizes conformational alterations of tau, which are associated with early neurofibrillary tangle formation. Pretreating sections with RNase or DNase-I showed immunostaining for both adducts was primarily associated with RNA. In addition, levels of both adducts in MCI were comparable to those measured in LAD, suggesting RNA oxidation may be an early event in the pathogenesis of neuron degeneration in AD. PMID- 17920287 TI - Different effect of unfractionated heparin and enoxaparin on circulating proangiogenic factors during hemodialysis: A cross-over study. AB - BACKGROUND: We aimed to compare the effect of unfractionated heparin (UFH) and enoxaparin used as anticoagulants during hemodialysis (HD) on circulating levels of heparin-binding, endothelial-derived, proangiogenic factors-vascular endothelial (VEGF(165)) and basic fibroblast (bFGF) growth factor. METHODS: We enrolled 22 chronic HD patients, who were randomly assigned to either enoxaparin (n=11) or UFH (n=11) anticoagulation, and followed prospectively for 12 weeks before crossing over to the alternate therapy for further 12 weeks. The cytokines were measured by immunoassay at the start, at 10 and 180min of HD. RESULTS: The baseline VEGF(165) and bFGF levels were comparable during enoxaparin and UFH treatment. VEGF(165) significantly decreased during both enoxaparin (chi(2) ANOVA=33.0, P<10(-6)) and UFH (chi(2) ANOVA=27.2, P<10(-6)) anticoagulated HD, while over-HD bFGF remained stable regardless of the type of heparin. The switch from enoxaparin to UFH treatment was connected with 34% VEGF(165) decrease after 180min of HD and had no impact on bFGF. During UFH-anticoagulated HD 75% VEGF(165) decrease after 10min was negatively associated with heparin dosage and was more profound in patients with ischemic heart disease. CONCLUSION: The traditional UFH regimen, in contrast to enoxaparin treatment, is connected with dose-depended VEGF(165) decrease during HD procedure. The biological and possible clinical relevance of this observation requires further investigations. PMID- 17920288 TI - Interaction characteristics of flavonoids with human organic anion transporter 1 (hOAT1) and 3 (hOAT3). AB - The present study aimed to investigate the interaction characteristics of flavonoids with human organic anion transporter 1 (hOAT1) and 3 (hOAT3). Five flavonoids (morin, silybin, naringin, naringenin and quercetin) were selected and their interaction characteristics with hOAT1 and hOAT3 were examined in MDCK cells overexpressing hOAT1 or hOAT3. Among tested flavonoids, morin and silybin exhibited significant inhibition effects on the cellular uptake of [3H]-para aminohippuric acid ([3H]-PAH) in MDCK-hOAT1 cells with Ki of 0.46 microM and 24 microM, respectively, while all the tested flavonoids appeared to be less interactive with hOAT3 compared to hOAT1. A kinetic study suggested that morin and silybin inhibited hOAT1-mediated cellular uptake of [3H]-PAH in a competitive manner. Furthermore, morin and silybin were translocated by hOAT1 across the cellular membrane. In conclusion, the present study identified some of flavonoids as a new class of hOAT1 inhibitors, suggesting a potential for flavonoid-drug interactions via the modulation of hOAT1 activity. PMID- 17920289 TI - Ammonia elimination from protonated nucleobases and related synthetic substrates. AB - The results are reported of mass-spectrometric studies of the nucleobases adenine 1h (1, R = H), guanine 2h, and cytosine 3h. The protonated nucleobases are generated by electrospray ionization of adenosine 1r (1, R = ribose), guanosine 2r, and deoxycytidine 3d (3, R = deoxyribose) and their fragmentations were studied with tandem mass spectrometry. In contrast to previous EI-MS studies of the nucleobases, NH(3) elimination does present a major path for the fragmentations of the ions [1h + H](+), [2h + H](+), and [3h + H](+). The ion [2h + H - NH(3)](+) also was generated from the acyclic precursor 5-cyanoamino-4 oxomethylene-dihydroimidazole 13h and from the thioether derivative 14h of 2h (NH(2) replaced by MeS). The analyses of the modes of initial fragmentation is supported by density functional theoretical studies. Conjugate acids 15-55 were studied to determine site preferences for the protonations of 1h, 2h, 3h, 13h, and 14h. The proton affinity of the amino group hardly ever is the substrate's best protonation site, and possible mechanisms for NH(3) elimination are discussed in which the amino group serves as the dissociative protonation site. The results provide semi-direct experimental evidence for the existence of the pyrimidine ring-opened cations that we had proposed on the basis of theoretical studies as intermediates in nitrosative nucleobase deamination. PMID- 17920290 TI - Top-down quantitation and characterization of SILAC-labeled proteins. AB - Stable isotope labeling by amino acids in cell culture (SILAC) has become a popular labeling strategy for peptide quantitation in proteomics experiments. If the SILAC technology could be extended to intact proteins, it would enable direct quantitation of their relative expression levels and of the degree of modification between different samples. Here we show through modeling and experiments that SILAC is suitable for intact protein quantitation and top-down characterization. When SILAC-labeling lysine and/or arginine, peaks of light and heavy SILAC-doublets do not interfere with peaks of different charge states at least between 10 and 200 kDa. Unlike chemical methods, SILAC ensures complete incorporation-all amino acids are labeled. The isotopic enrichment of commercially available SILAC amino acids of nominally 95% to 98% shifts the mass difference between light and heavy state but does not lead to appreciably broadened peaks. We expressed labeled and unlabeled Grb2, a 28 kDa signaling protein, and showed that the two forms can be quantified with an average standard deviation of 6%. We performed on-line top-down sequencing of both forms in a hybrid linear ion trap orbitrap instrument. The quantized mass offset between fragments provided information about the number of labeled residues in the fragments, thereby simplifying protein identification and characterization. PMID- 17920291 TI - Study of Michael-Michael-retro Michael addition catalyzed by 9-amino-9 deoxyepiquinine using ESI-MS. AB - The Michael-Michael-retro Michael addition catalyzed by 9-amino-9-deoxyepiquinine was monitored and the major intermediates and catalyst in the catalytic cycle were detected and characterized using ESI-MS/MS for the first time. Some important isomeric intermediates including isomeric enamine and imine structures were tentatively differentiated and further studied by theoretical calculations. Fragment ions of protonated catalyst indicate that proton possibly influences the conformation of the catalyst. PMID- 17920293 TI - Progenitor cell maintenance and neurogenesis in sympathetic ganglia involves Notch signaling. AB - Differentiation of noradrenergic neurons from neural crest-derived precursors results in the formation of primary sympathetic ganglia. As sympathetic neurons continue to divide after the acquisition of adrenergic and neuronal properties it was unclear, whether the increase in neuron number during neurogenesis is due to neuron proliferation rather than differentiation of progenitor cells. Here, we demonstrate Sox10-positive neural crest progenitor cells and continuous sympathetic neuron generation from Phox2b-positive autonomic progenitors during early chick sympathetic ganglion development. In vivo activation of Notch signaling resulted in a decreased neuronal population, whereas expression of the Notch signaling inhibitor Su(H)(DBM) increased the proportion of Scg10-positive neurons. Similar results were obtained for sensory dorsal root ganglia (DRG). The effects of Notch gain- and loss-of-function experiments support the notion that progenitor maintenance and neuron differentiation from progenitor cells are essential for neurogenesis also during early sympathetic ganglion development. PMID- 17920294 TI - Glycinergic input of small-field amacrine cells in the retinas of wildtype and glycine receptor deficient mice. AB - Amacrine cells are known to express strychnine-sensitive glycine receptors (GlyRs), however, it is not known which of the four GlyRalpha subunits (alpha1-4) are expressed in this diverse group of cells. Herein, we studied the presence of glycine activated currents and spontaneous inhibitory postsynaptic currents (sIPSCs) of amacrine cells in the mouse retina. By recording glycinergic currents in retinal slices of wildtype mice and of mice deficient in GlyRalpha subunits (Glra1spd-ot, Glra2-/-, Glra3-/-), we could classify AII and narrow-field amacrine cells (NF, Types 5, 6, 7) on the basis of their alpha-subunit composition. Glycinergic sIPSCs of AII cells displayed medium fast kinetics (mean decay time constant tau=11+/-2 ms), which were completely absent in the Glra3-/- mouse, indicating that synaptic GlyRs of AII cells mainly contain the alpha3 subunit. Glycinergic sIPSCs of NF cells had slow kinetics (tau=27+/-6.8 ms) that were significantly prolonged in Glra2-/- mice (tau=69+/-16 ms). These data show that morphologically distinct amacrine cells express different sets of GlyRs. PMID- 17920292 TI - Heterogeneous nuclear ribonucleoprotein A/B and G inhibits the transcription of gonadotropin-releasing-hormone 1. AB - Gonadotropin-releasing hormone 1 (GnRH1) causes the release of gonadotropins from the pituitary to control reproduction. Here we report that two heterogeneous nuclear ribonucleoproteins (hnRNP-A/B and hnRNP-G) bind to the GnRH-I upstream promoter region in a cichlid fish Astatotilapia burtoni. We identified these binding proteins using a newly developed homology based method of mass spectrometric peptide mapping. We show that both hnRNP-A/B and hnRNP-G co localize with GnRH1 in the pre-optic area of the hypothalamus in the brain. We also demonstrated that these ribonucleoproteins exhibit similar binding capacity in vivo, using immortalized mouse GT1-7 cells where overexpression of either hnRNP-A/B or hnRNP-G significantly down-regulates GnRH1 mRNA levels in GT1-7 cells, suggesting that both act as repressors in GnRH1 transcriptional regulation. PMID- 17920295 TI - Expression and characterization of a pleckstrin homology domain in phospholipase C, PLC-eta1. AB - Phospholipase C (PLC) plays an important role in intracellular signal transduction by hydrolyzing phosphatidylinositol 4,5-bis-phosphate, a membrane phospholipid. Currently, thirteen mammalian PLC isozymes have been identified, which are divided into six classes on the basis of structure and mechanisms. All the PLC isozymes share common domains including catalytic X and Y domains, protein kinase C conserved region 2 (C2) domain, EF-hand motif and pleckstrin homology (PH) domain. In this study, the PLC-eta1 PH domain has been over expressed and purified. The most undesirable feature of the protein was instability, resulting in precipitation during the purification process. With the aim of structural characterization, a solution condition was optimized using SDS PAGE and NMR spectroscopy. A circular dichroism spectrum indicated that the PLC eta1 PH domain mainly comprised beta-strands, which was also suggested by the 2D 1H-15N HSQC spectrum. PMID- 17920296 TI - Toward direct neural current imaging by resonant mechanisms at ultra-low field. AB - A variety of techniques have been developed to noninvasively image human brain function that are central to research and clinical applications endeavoring to understand how the brain works and to detect pathology (e.g. epilepsy, schizophrenia, etc.). Current methods can be broadly divided into those that rely on hemodynamic responses as indicators of neural activity (e.g. fMRI, optical, and PET) and methods that measure neural activity directly (e.g. MEG and EEG). The approaches all suffer from poor temporal resolution, poor spatial localization, or indirectly measuring neural activity. It has been suggested that the proton spin population will be altered by neural activity resulting in a measurable effect on the NMR signal that can be imaged by MRI methods. We present here the physical basis and experimental evidence for the resonant interaction between magnetic fields such as those arising from neural activity, with the spin population in ultra-low field (microT) NMR experiments. We demonstrate through the use of current phantoms that, in the case of correlated zero-mean current distributions such as those one might expect to result from neural activity, resonant interactions will produce larger changes in the observed NMR signal than dephasing. The observed resonant interactions reported here might one day form the foundation of a new functional neuroimaging modality ultimately capable of simultaneous direct neural activity and brain anatomy tomography. PMID- 17920297 TI - BOLD and perfusion changes during epileptic generalised spike wave activity. AB - It is unclear whether neurovascular coupling is maintained during epileptic discharges. Knowing this is important to allow appropriate inferences from functional imaging studies of epileptic activity. Recent blood oxygen level dependent (BOLD) functional MRI (fMRI) studies have demonstrated negative BOLD responses (NBR) in frontal, parietal and posterior cingulate cortices during generalised spike wave activity (GSW). We hypothesized that GSW-related NBR commonly reflect decreased cerebral blood flow (CBF). We measured BOLD and cerebral blood flow responses using simultaneous EEG with BOLD and arterial spin label (ASL) fMRI at 3 T. Four patients with epilepsy were studied; two with idiopathic generalized epilepsy (IGE) and two with secondary generalized epilepsy (SGE). We found GSW-related NBR in frontal, parietal and posterior cingulate cortices. We measured the coupling between BOLD and CBF changes during GSW and normal background EEG and found a positive correlation between the simultaneously measured BOLD and CBF throughout the imaged volume. Frontal and thalamic activation were seen in two patients with SGE, concordant with the electro clinical features of their epilepsy. There was striking reproducibility of the GSW-associated BOLD response in subjects previously studied at 1.5 T. Our results show a preserved relationship between BOLD and CBF changes during rest and GSW activity consistent with normal neurovascular coupling in patients with generalized epilepsy and in particular during GSW activity. Cortical activations appear to reflect areas of discharge generation whilst deactivations reflect changes in conscious resting state activity. PMID- 17920298 TI - Defining a monophyletic Cardinalini: a molecular perspective. AB - Within the New World nine-primaried oscine assemblage, feeding morphology and behavior have long been used as a guideline for assigning membership to subgroups. For example, birds with stout, conical bills capable of crushing heavy seeds have generally been placed within the tribe Cardinalini (cardinal grosbeaks). Many workers have tried to characterize this group more definitively, using a variety of morphological characters; however, the characters used often conflicted with one another. Previous molecular studies addressing the monophyly of Cardinalini have had only limited sampling within the group. In this study, we analyze mtDNA sequence data from all genera and 34 of the 42 Cardinalini species (sensu [Sibley, C.G., Monroe, B.L., 1990. Distribution and Taxonomy of the Birds of the World, Yale University Press, New Haven, CT]) to address the monophyly of the group and to reconstruct the most complete phylogeny of this tribe published to date. We found strong support for a redefined Cardinalini that now includes some members previously placed within Thraupini (tanagers; the genera Piranga, Habia, Chlorothraupis, and Amaurospiza) and some members previously placed within the Parulini (wood-warblers; the genus Granatellus). In addition, some genera traditionally considered members of the Cardinalini are shown to have affinities with other groups (the genera Porphyrospiza, Parkerthraustes, and Saltator). Our redefined Cardinalini contains 48 species, six more than are listed in Sibley and Monroe's (1990) taxonomy of the group. Within the nine-primaried oscine assemblage, the Cardinalini are more closely related to the Thraupini (tanagers) than they are to the Emberizini (sparrows), Parulini (wood-warblers), or Icterini (blackbirds), consistently forming a monophyletic group with Thraupini across all analyses. The reconfigured Cardinalini is comprised of five well-supported, major clades: (1) a "masked" clade (Piranga, Cardinalis, Caryothraustes, Periporphyrus, and Rhodothraupis), (2) a "blue" clade (Amaurospiza, Cyanocompsa, Cyanoloxia, Passerina, and Spiza), (3) a clade containing the genera Habia and Chlorothraupis, (4) a clade containing all species of Granatellus, and (5) a clade containing only species of Pheucticus. Diversification of these five lineages from one another occurred relatively rapidly during the mid-Pliocene, around 5 or 8 million years ago. Each of these major clades includes both North and South American species; thus, a complex biogeographic history is inferred for the group. PMID- 17920299 TI - Geographical patterns of deep mitochondrial differentiation in widespread Malagasy reptiles. AB - Using sequences of the mitochondrial 16S rRNA gene, we reconstructed the phylogeography of six widely distributed Malagasy reptiles: two gekkonid lizard species, Phelsuma lineata and Hemidactylus mercatorius; two chameleons, the Calumma brevicorne complex, and Furcifer lateralis; and two skinks, Trachylepis gravenhorstii and Trachylepis elegans. Genetic differentiation among major haplotype lineages was high and in some cases indicates or confirms species status of the divergent populations. Maximum uncorrected sequence divergences were between 2.2% and 8.3% within the various species or species complexes. Haplotype lineages were exclusive to geographic regions, except in the commensal H. mercatorius where in three anthropogenic habitats coexistence of haplotype lineages was observed, possibly due to human translocation. The eastward flowing rivers Mangoro and Mananara may represent barriers to gene flow in the case of three species each. Some species sampled from humid eastern and arid western Madagascar showed no differentiation between populations from these two regions; instead the pattern observed was in several cases more concordant with a differentiation along a north-south axis. PMID- 17920300 TI - Phylogeographic patterns in widespread corvid birds. AB - Intraspecific genetic diversity and phylogeography of Corvus corone was investigated using the mitochondrial (mt) control region as a molecular marker. A split into two distinct mt lineages was observed. One represents individuals from a wide geographic range spanning from England to the Russian Far East (Kamchatka), while the other one was found in the Primorye and Khabarovsk regions (southern parts of Russian Far East) as well as Japan. For comparison, we investigated several widespread Palearctic corvid taxa with respect to their phylogeographic patterns. A deep split into two lineages was revealed in five cases: Besides C. corone, within Corvus frugilegus, Pica pica, and between the species pairs Corvus monedula-Corvus dauuricus and Cyanopica cyanus-Cyanopica cooki. Although these taxa display a variety of distribution patterns, from disjunct, para/allopatric to continuous, the genetic pattern and level of divergence between clades is very similar. This implies that the differentiation started in about the same time range. In contrast, no differentiation into highly divergent lineages was detected in Corvus corax, Perisoreus infaustus, and Nucifraga caryocatactes. We try to explain the two phylogeographic patterns in corvid birds with ecological factors accompanying the changing climatic conditions during the Pleistocene. The deep genetic splits within several widely distributed Palearctic corvids are discussed with respect to taxonomic questions. PMID- 17920301 TI - Assessing phylogenetic resolution among mitochondrial, nuclear, and morphological datasets in Nothonotus darters (Teleostei: Percidae). AB - External morphological characters are the basis of our understanding of diversity and species relationships in many darter clades. The past decade has seen the publication of many studies utilizing mtDNA sequence data to investigate darter phylogenetics, but only recently have nuclear genes been used to investigate darter relationships. Despite a long tradition of use in darter systematics few studies have examined the phylogenetic utility of external morphological characters in estimating relationships among species in darter clades. We present DNA sequence data from the mitochondrial cytochrome b (cytb) gene, the nuclear encoded S7 intron 1, and discretely coded external morphological characters for all 20 species in the darter clade Nothonotus. Bayesian phylogenetic analyses result in phylogenies that are in broad agreement with previous studies. The cytb gene tree is well resolved, while the nuclear S7 gene tree lacks phylogenetic resolution, node support, and is characterized by a lack of reciprocal monophyly for many of the Nothonotus species. The phylogenies resulting from analysis of the morphological dataset lack resolution, but nodes present are found in the cytb and S7 gene trees. The highest resolution and node support is found in the Bayesian combined data phylogeny. Based on our results we propose continued exploration of the phylogenetic utility of external morphological characters in other darter clades. Given the extensive lack of reciprocal monophyly of species observed in the S7 gene tree we predict that nuclear gene sequences may have limited utility in intraspecific phylogeographic studies of Nothonotus darters. PMID- 17920302 TI - Effect of order of presentation of a generic and a specific health-related quality of life instrument in knee and hip osteoarthritis: a randomized study. AB - OBJECTIVE: Current guidelines recommend using both a generic and a specific instrument to measure quality of life (QoL) among people with chronic diseases. However, the two questionnaires may not be independent, which raises the issue of whether the order in which they are completed influences their value. We aimed to assess the effect of order of presentation of a generic (SF36) and a specific (Osteoarthritis Knee and Hip QoL [OAKHQOL]) QoL instrument administered to patients with knee and hip osteoarthritis (OA). METHODS: We recruited 341 outpatients from rheumatology and orthopaedic surgery clinics. Demographic and clinical data and responses to the SF36 and OAKHQOL self-administered questionnaires were collected at baseline and 10 days after inclusion; surgical patients were also assessed 6 months post-surgery. The order of presentation of the instruments was randomized at inclusion. RESULTS: The order of instrument presentation had no significant effect on response rate, number of dimensions with a floor or a ceiling effect, or questionnaire scores. In one of 13 dimensions (social support dimension of the OAKHQOL), test-retest reliability was slightly better when the generic SF36 questionnaire was presented first (intra class correlation coefficient (ICC) 0.84 vs 0.55). The analysis of sensitivity to change and discriminant ability did not favour one group over the other. CONCLUSION: The order of presentation of a generic and a specific QoL questionnaire to patients with OA had an only marginal effect on the quality of responses and the QoL scores obtained. PMID- 17920303 TI - Rhabdastrellic acid-A inhibited PI3K/Akt pathway and induced apoptosis in human leukemia HL-60 cells. AB - Increasing evidence suggests that aberrant activation of PI3K/Akt is involved in many human cancers, and that inhibition of the PI3K/Akt pathway might be a promising strategy for cancer treatment. Our investigation indicates that Rhabdastrellic acid-A, an isomalabaricane triterpenoid isolated from the sponge, Rhabdastrella globostellata, inhibits proliferation of HL-60 cells with an IC(50) value of 0.68mug/ml, and induces apoptosis. Rhabdastrellic acid-A also induces cleavage of the death substrate poly (ADP-ribose) polymerase (PARP) and caspase 3. Pretreatment of HL-60 cells with the caspase-3 specific inhibitor, DEVD-CHO, prevents Rhabdastrellic acid-A-induced DNA fragmentation and PARP cleavage. Activated PI3K and Akt significantly decreases after treatment with Rhabdastrellic acid-A in HL-60 cells. Expression levels of protein bcl-2, bax remain unchanged in response to Rhabdastrellic acid-A treatment in HL-60 cells. These results suggest that Rhabdastrellic acid-A inhibits PI3K/Akt pathway and induces caspase-3 dependent-apoptosis in HL-60 human leukemia cells. PMID- 17920304 TI - Effect on memory of acute administration of naturally secreted fibrils and synthetic amyloid-beta peptides in an invertebrate model. AB - Amyloid beta peptide (Abeta) is considered one of the main agents of Alzheimer's disease pathogenesis. Recently, it has been proposed that memory deficits are caused by different stages of Abeta aggregation, particularly by oligomers. In addition, although memory impairment was found after Abeta administration in rodents and chicks, the nature of the memory deficits induced in invertebrates by acute administration of mammalian Abeta peptides is not well understood. Previously, we reported the amnesic effect of acute pre-training administration of naturally formed fibrils (NF) in crab memory. Here we evaluate the effect of NF and synthetic Abeta peptides administration at different times before and after training in this well characterized invertebrate memory model, the context signal memory of the crab Chasmagnathus. We found a clear amnesic effect at very low doses of naturally Abeta NF only when administered immediately pre- and post training, but not 24 h and 18 h before or 6h after training. Activation of ERK/MAPK (a protein kinase required for memory formation in this model) 60 min after administration was found. In contrast, neither JNK/SAPK nor NF-kappaB transcription factor were activated. Furthermore, synthetic Abeta1-42 and Abetapy3-42 administration induced amnesia when used after a protocol for fibrillation but not after a protocol for oligomerization. On the contrary, no amnestic effect was found when fibrillated Abeta1-40 and Abetapy11-42 peptides were used. Thus, Abeta1-42 and Abetapy3-42 peptides impaired memory and the effects were only found when highly aggregated peptides, which may include fibrils, protofibrils and oligomers, were administered. These temporally- and signaling-specific effects suggest that Abeta impairs memory by inducing transient physiological, rather than permanent neuropathological, alterations of the brain and this effect is achieved through generalized ERK activation. PMID- 17920305 TI - Relationship of preoperative venous and arterial imaging findings to outcomes of brachio-basilic transposition fistulae for hemodialysis: a prospective clinical study. AB - PURPOSE: To evaluate the maturation and patency of transposed brachio-basilic fistulae that had been chosen based on the results of preoperative imaging techniques. METHODS: Among 215 patients admitted to our clinic requiring arteriovenous fistulae between May 2004 and September 2005, 59 were scheduled for a transposed brachio-basilic fistula procedure. The relationship between demographic data, laboratory values, invasive and noninvasive imaging studies with patency and maturation time of the fistulae were evaluated by univariate analyses. RESULTS: Primary and secondary patency rates were 82% and 97% at 6 months 72% and 92% at one year respectively. The only parameter found to affect maturation time was basilic vein diameter. The maturation time was 59.3+/-22.3 days (range 32-92 days) for veins less than 3mm in diameter and 24.7+/-4.4 days (range, 21 to 34 days) for those with larger diameters. The number of previously failed fistulae correlated with a decrease in primary patency time. The primary patency rate at 1 year was 58.9% for patients whose preoperative arterial flow rate was below 70cm/sec, while it was 93.3% when the flow was greater. CONCLUSION: We believe that this type of fistula should be the first option in patients in whom the cephalic vein is inappropriate for a vascular access. Preoperative evaluation of the arterial system as well as a history of previous access failure may be considered predictive parameters for the patency of the fistulae. PMID- 17920306 TI - Superficial femoral artery recanalization with self-expanding nitinol stents: long-term follow-up results. AB - PURPOSE: Since long-term patency and device integrity of nitinol stents in SFA lesions are not well studied, we examined clinical outcome, patency and device integrity after stenting long lesions using a standardized implantation technique. METHODS: Between 2001 and 2006, 59 patients (74 lesions) were treated with the same nitinol self-expandable stent (Zilver, Cook, USA) and technique for SFA recanalization. Clinical charts and imaging were retrospectively reviewed for patency (primary and assisted-primary), and device integrity. RESULTS: Patients were 74.5 (10.9) years old (range 49 to 93), 64% male, 42% diabetic, 62% hypertensive and 67% current or former smokers. Lesions were 23% TASC B, 16% TASC C, or 61% TASC D. Mean recanalization length was 19 cm (range 3 to 53). Mean number of stents per patient was 2.8 (total 210). Mean follow-up time was 2.4 years (range 3 days to 4.8 years). Kaplan-Meier estimates for primary patency rates were 90%, 78%, 74%, 69%, and 69% at 1, 2, 3, 4 and 4.8 years, respectively. Ten restenoses at a mean of 500 (388) days (1-1251 days) were successfully recanalized. The assisted primary patency rates were 96%, 90%, 90%, 90% and 90% at 1, 2, 3, 4 and 5 years, respectively. Six complete occlusions could not be reverted by a second recanalization procedure, and were treated by surgical bypass (1 case), amputation (3 cases), or medical management (2 cases). One (1.04%) Class II stent fracture was noted. CONCLUSIONS: SFA recanalization with a standardized implantation technique and nitinol stents provides good long-term primary and assisted-primary patency. PMID- 17920307 TI - Technical review of endovenous laser therapy for varicose veins. AB - BACKGROUND: In the last decade, several new treatments of truncal varicose veins have been introduced. Of these new therapies, endovenous laser therapy (EVLT) is one of the most widely accepted and used treatment options for incompetent greater and lesser saphenous veins. OBJECTIVE: The objective of this report is to inform clinicians about the EVLT procedure and to review its efficacy and safety in treatment of truncal varicose veins. Also, we discuss some of the underlying theoretical principles and laser parameters that affect EVLT. METHODS: We carried out a literature review of EVLT;s efficacy and safety. We included reports that included 100 or more limbs with a follow-up of at least 3 months. The principals and procedure of EVLT are described. Of the laser parameters, mode of administration, wavelength, fluence, wattage and pullback speed are discussed. CONCLUSION: EVLT appears to be a very effective and safe option in the treatment of varicose veins but large randomized comparative studies are needed. PMID- 17920309 TI - Paraplegia following elective endovascular repair of abdominal aortic aneurysm: reversal with cerebrospinal fluid drainage. AB - Paraplegia secondary to spinal cord ischaemia is a rare but devastating complication of abdominal aortic aneurysm repair. We report a case of paraplegia following elective endovascular repair of an infrarenal aortic aneurysm. A cerebrospinal fluid (CSF) drain was immediately inserted and resulted in full neurological recovery. This case highlights the fact that endovascular techniques are prone to similar complications as open surgery, and the importance of prompt cerebrospinal fluid drainage in cases of spinal cord ischaemia. PMID- 17920308 TI - The nature of skin pigmentations in chronic venous insufficiency: a preliminary report. AB - BACKGROUND: Chronic Venous Insufficiency (CVI) provokes skin pigmentation commonly seen in the gaiter region of the leg. The exact nature and pathogenesis of this are poorly understood. OBJECTIVE: To evaluate the presence of melanin and haemosiderin in histological sections of the skin of limbs with primary varicose veins. METHODS: Histological investigations were performed in 49 biopsies from pigmented and non-pigmented skin of limbs with varicose veins and control limbs. RESULTS: All samples from pigmented skin showed a higher content of melanin than controls. In contrast, haemosiderin was found in only a few biopsies taken from the more severely pigmented skin in areas of lipodermatosclerosis. Erythrocyte diapedesis was observed only where an intense inflammatory process was also present. CONCLUSIONS: Our findings suggest that in the initial phases of skin changes due to venous disease, pigmentation is attributable to melanin. Haemosiderin seems to play a role in the evolution of skin changes toward lipodermatosclerosis and ulceration. Erythrocyte diapedesis is likely to occur only during acute phases of the inflammatory process. Further investigations are needed to explain the cause and the exact cellular and molecular mechanisms responsible for hypermelanisation occurring in early phases of skin changes in CVI. PMID- 17920310 TI - Activated coagulation in patients with shock due to ruptured abdominal aortic aneurysm. AB - BACKGROUND: Ruptured abdominal aortic aneurysm is associated with a high operative mortality. Postoperative thrombosis related complications are common, a possible mechanism being activation of the coagulation system and endothelial stimulation. The aim of the present study was to investigate the coagulation activity preoperatively in patients with ruptured and nonruptured abdominal aortic aneurysm in relation to the clinical outcome with special regard to the influence of shock. METHODS: Ninety-five patients with repair of infrarenal aortic aneurysm and forty-one controls without aneurysm matched by age, gender and smoking habits were studied. Thrombin-antithrombin (TAT), prothrombin fragment 1+2 (F 1+2), and von Willebrand factor antigen (vWFAg) were measured. RESULTS: There were significantly higher levels of TAT, F 1+2, and vWFAg in patients operated for ruptured compared to nonruptured abdominal aortic aneurysm. The highest level of TAT and F 1+2 were detected in patients with rupture and shock. CONCLUSION: The present data indicate a state of activated coagulation in patients with ruptured abdominal aortic aneurysm which is reinforced by shock. PMID- 17920311 TI - Candidate gene association studies in abdominal aortic aneurysm disease: a review and meta-analysis. AB - BACKGROUND: Candidate gene analysis has been frequently used in attempts to understand the pathological processes involved in many aspects of AAA disease. METHODS: This paper sets out a systems approach to reviewing AAA candidate gene analysis studies, whilst, explaining the key principles and design limitations of this universally applied technique. In addition we have performed a meta-analysis of six gene polymorphisms (ACE I/D, MTHFR+677C>T, MMP9-1562C>T, Il-1Beta/3953C>T, eNOS 4a/4b & TIMP1/+434C>T) reported in multiple case control studies. RESULTS AND CONCLUSIONS: Three of these polymorphisms were associated with a significant risk of AAA, ACE RR 1.33 [95% CI 1.20-1.48], MTHFR RR 1.14 [1.08-1.21] and MMP9 RR 1.09 [1.01-1.18]. These differences have been previously reported as equivocal, within a context of contradictory studies and as such this meta analysis provides new evidence for their involvement in AAA disease. The plausibility of these findings is discussed within the context of a systems approach to the pathology of AAA disease. PMID- 17920312 TI - Gatekeeper function of the RUNX1 transcription factor in acute leukemia. AB - The RUNX1 gene encodes the alpha subunit of the core binding factor (CBF) and is a common target of genetic mutations in acute leukemia. We propose that RUNX1 is a gatekeeper gene, the disruption of which leads to the exodus of a subset of hematopoietic progenitors with increased self-renewal potential from the normal environmental controls of homeostasis. This pool of "escaped" cells is the target of secondary mutations, accumulating over time to induce the aggressive manifestation of acute leukemia. Evidence from patient and animal studies supports the concept that RUNX1 mutations are the initiating event in different leukemia subtypes, but also suggests that diverse mechanisms are used to subvert RUNX1 function. One common result is the inhibition of differentiation-but its effect impinges on different lineages and stages of differentiation, depending on the mutation or fusion partner. A number of different approaches have led to the identification of secondary events that lead to the overt acute phase; however, the majority is unknown. Finally, the concept of the "leukemia stem cell" and its therapeutic importance is discussed in light of the RUNX1 gatekeeper function. PMID- 17920313 TI - The perichromatin region: a functional compartment in the nucleus that determines large-scale chromatin folding. AB - The perichromatin region has emerged as an important functional domain of the interphase nucleus. Major nuclear functions, such as DNA replication and transcription, as well as different RNA processing factors, occur within this domain. In this review, we summarize in situ observations regarding chromatin structure analysed by transmission electron microscopy and compare results to data obtained by other methods. In particular, we address the functional architecture of the perichromatin region and the way chromatin may be folded within this nucleoplasmic domain. PMID- 17920314 TI - Role of Ace2 (Activator of Cellulases 2) within the xyn2 transcriptosome of Hypocrea jecorina. AB - Ace2 (Activator of Cellulases 2)-encoding gene was deleted from and retransformed in the H. jecorinaQM9414 genome. Comparison of xylanase activity and xyn2 transcription of the corresponding strains after cultivation on inducing compounds (xylan, xylobiose) revealed a faster initial inducibility in the Deltaace2-strain, but final levels of xyn2 transcript and xylanase activity of the parental strain could not be reached. This suggests a role for Ace2 in the regulation of xyn2 induction mechanisms, moreover Ace2 is responsible for the basal level of xyn2 transcription. Furthermore, a palindrome in the xyn2 promoter consisting of a GGGTAA- and a CCAGCC-element was identified. Both Xyr1 and Ace2 are able to bind the complete motif, the latter also only to one part of it. Phosphorylation as well as dimerization are prerequisites for binding of Ace2 to the xyn2 promoter. Finally, the impact of Ace2 on xyr1 transcription could be demonstrated under inducing conditions. PMID- 17920315 TI - Changes in protein structure and dynamics as a function of hydration from (1)H second moments. AB - We report the proton second moment obtained directly from the Free Induction Decay (FID) of the NMR signal of variously hydrated bovine serum albumin (BSA) and hen egg white lysozyme (HEWL) and from the width of the NMR Z-spectrum of the cross-linked protein gels of different concentrations. The second moment of the proteins decreases in a continuous stepwise way as a function of increasing water content, which suggests that the structural and dynamical changes occur in small incremental steps. Although the second moment is dominated by the short range distances of nearest neighbors, the changes in the second moment show that the protein structure becomes more open with increasing hydration level. A difference between the apparent liquid content of the sample as found from decomposition of the FID and the analytically determined water content demonstrates that water absorbed in the early stages of hydration is motionally immobilized and magnetically indistinguishable from rigid protein protons while at high hydration levels some protein side-chain protons move rapidly contributing to liquid-like component of the NMR signal. PMID- 17920316 TI - Low-E probe for (19)F-(1)H NMR of dilute biological solids. AB - Sample heating induced by radio frequency (RF) irradiation presents a significant challenge to solid state NMR experiments in proteins and other biological systems, causing the sample to dehydrate which may result in distorted spectra and a damaged sample. In this work we describe a large volume, low-E (19)F-(1)H solid state NMR probe, which we developed for the 2D (19)F CPMG studies of dilute membrane proteins in a static and electrically lossy environment at 600MHz field. In (19)FCPMG and related multi-pulse (19)F-(1)H experiments the sample is heated by the conservative electric fields E produced in the sample coil at both (19)F and (1)H frequencies. Instead of using a traditional sample solenoid, our low-E (19)F-(1)H probe utilizes two orthogonal loop-gap resonators in order to minimize the conservative electric fields responsible for sample heating. Absence of the wavelength effects in loop-gap resonators results in homogeneous RF fields and enables the study of large sample volumes, an important feature for the dilute protein preparations. The orthogonal resonators also provide intrinsic isolation between the (19)F and (1)H channels, which is another major challenge for the (19)F-(1)H circuits where Larmor frequencies are only 6% apart. We detail steps to reduce (19)F background signals from the probe, which included careful choice of capacitor lubricants and manufacture of custom non-fluorinated coaxial cables. Application of the probe for two-dimensional (19)F CPMG spectroscopy in oriented lipid membranes is demonstrated with Flufenamic acid (FFA), a non-steroidal anti inflammatory drug. PMID- 17920317 TI - Removal of t(1) noise from metabolomic 2D (1)H-(13)C HSQC NMR spectra by Correlated Trace Denoising. AB - The presence of t(1) noise artefacts in 2D phase-cycled Heteronuclear Single Quantum Coherence (HSQC) spectra constrains the use of this experiment despite its superior sensitivity. This paper proposes a new processing algorithm, working in the frequency-domain, for reducing t(1) noise. The algorithm has been developed for use in contexts, such as metabolomic studies, where existing denoising techniques cannot always be applied. Two test cases are presented that show the algorithm to be effective in improving the SNR of peaks embedded within t(1) noise by a factor of more than 2, while retaining the intensity and shape of genuine peaks. PMID- 17920318 TI - Optical coherence tomography as a tool for monitoring pediatric pseudotumor cerebri. AB - PURPOSE: To evaluate optical coherence tomography (OCT) measurement of peripapillary retinal nerve fiber layer (RNFL) and macular thickness in children with pseudotumor cerebri. METHODS: Prospective observational series of children with pseudotumor cerebri compared against controls matched for age and gender. We included 11 pediatric subjects with clinical pseudotumor cerebri and 37 normal controls. Subjects underwent a complete eye examination including stereo disk photographs and OCT. Peripapillary RNFL and macular thickness were evaluated using the Stratus OCT 3000. The peripapillary RNFL was evaluated with two protocols: Fast RNFL Map protocol with values divided in eight sections, made up of four quadrants, each with an inner and outer ring, and Fast RNFL Thickness (3.4) protocol, using values presented for four quadrants. A Fast Macular Thickness Map was also obtained, and results reported similarly included eight sections, made up of four quadrants, each with an inner and outer ring. RESULTS: The Fast RNFL Map protocol showed an increased RNFL thickness (pseudotumor cerebri vs controls) in the temporal and superior quadrants. The Fast RNFL Thickness (3.4) protocol showed a thicker average RNFL for pseudotumor cerebri eyes compared with controls (125.7 vs 106.5 microm, p < 0.0001). The Fast Macular Thickness Map showed an increased RNFL thickness in the nasal quadrant for the inner ring. The macular volumes for pseudotumor cerebri versus control eyes were 7.21 versus 6.97 mm(3), respectively (p = 0.005). CONCLUSIONS: OCT shows increased RNFL and macular thickness in pseudotumor cerebri and may be a useful clinical tool in these children. PMID- 17920319 TI - A comparison of hang-back with conventional recession surgery for exotropia. AB - BACKGROUND: The hang-back loop suspension surgical technique for rectus muscle recession offers potential advantages over the conventional rectus muscle recession, including better exposure at the site of scleral sutures, shorter procedure duration, and lower risk of scleral perforation. Previous reports suggest that the hang-back technique for lateral rectus recession for exotropia results in poorer surgical success and may require a different surgical dosage. METHODS: We compared strabismus in a nonrandomized series of 55 children with exotropia treated with conventional surgery or hang-back surgery. Those in the hang-back (suspension) recession group were on average 2.5 years younger and had 8(Delta) more preoperative exotropia at distance. RESULTS: Surgical outcomes were not significantly different in the two treatment groups. Multivariate logistic regression suggests that outcomes are at least equivalent to conventional surgery after adjusting for age, severity, and other preoperative factors likely associated with outcome. CONCLUSIONS: The hang-back surgical technique was as effective as conventional lateral rectus recession surgery for children with exotropia. PMID- 17920321 TI - The venous circulation: a piscine perspective. AB - Vascular capacitance describes the pressure-volume relationship of the circulatory system. The venous vasculature, which is the main capacitive region in the circulation, is actively controlled by various neurohumoral systems. In terrestrial animals, vascular capacitance control is crucial to prevent orthostatic blood pooling in dependent limbs, while in aquatic animals like fish, the effects of gravity are cancelled out by hydrostatic forces making orthostatic blood pooling an unlikely concern for these animals. Nevertheless, changes in venous capacitance have important implications on cardiovascular homeostasis in fish since it affects venous return and cardiac filling pressure (i.e. central venous blood pressure), which in turn may affect cardiac output. The mean circulatory filling pressure is used to estimate vascular capacitance. In unanaesthetized animals, it is measured as the central venous plateau pressure during a transient stoppage of cardiac output. So far, most studies of venous function in fish have addressed the situation in teleosts (notably the rainbow trout, Oncorhynchus mykiss), while any information on elasmobranchs, cyclostomes and air-breathing fishes is more limited. This review describes venous haemodynamic concepts and neurohumoral control systems in fish. Particular emphasis is placed on venous responses to natural cardiovascular challenges such as exercise, environmental hypoxia and temperature changes. PMID- 17920320 TI - Ocular cysticercosis of the anterior segment. AB - A 6-year-old girl presented with signs of severe anterior uveitis. After initiating treatment, a cyst like lesion was observed in the anterior chamber that led to the diagnosis of ocular cysticercosis that was ultimately confirmed with histopathologic analysis. Ocular cysticercosis usually affects the extraocular muscle. Infection of the anterior chamber has been described, although to a lesser extent. Because of the usually poor visual prognosis and the controversy in treatment, physicians should be aware of this disease and its different variants of presentation. PMID- 17920322 TI - Fractures and handicap in an adult population: A clinical study. AB - OBJECTIVE: To evaluate the bone status of ambulatory patients with physical and mental handicaps before a program of fracture prevention. METHODS: We recruited 58 walking adults. We retrospectively collected the past episodes of fractures, essentially peripheral, and epilepsy. The serum calcium, albumin, 25 hydroxyvitamin D, parathormone, CTX-1 and P1NP levels were prospectively measured in 36 consecutive patients. Each patient received daily calcium and vitamin D. The vertebral status has been not evaluated. RESULTS: Twenty-one patients had presented at least one fracture. Thirty nine per cent of the fractures were minor (nasal bone, hands, feet). The age of patients with fractures was significantly higher than patients without fracture (46 versus 40years, respectively; p=0.04). Patients with fractures had a significantly increased S-P1NP (63.5ng/ml+/-32.0 versus 41.9ng/ml+/-20.0, respectively; p=0.02). Nineteen patients suffered from epilepsy. We listed 23 fractures among 9 patients treated by phenobarbital and 8 fractures, which tended to be less severe among 5 patients epileptics without this drug. Minor fracture was often followed by severe fracture in case of phenobarbital treatment. This treatment was associated with a significantly lower serum calcium level (2.16mmol/l+/-0.05, versus epileptic patients without phenobarbital 2.32mmol/l+/-0.08, p<0.0004). CONCLUSIONS: The presence of a fracture, even minor, must encourage to improve the preventive and curative measures among patients with handicaps. PMID- 17920323 TI - Periosteal reaction in systemic lupus erythematosus. AB - Musculoskeletal complaints are the most common presenting symptoms in most of the patients with systemic lupus erythematosus (SLE). However, periosteal new bone formation is an extraordinarily rare condition in SLE. We report a case of periosteal reaction in SLE. A 31-year-old woman with SLE presented with both knee pain. Radiographs revealed periosteal reactions in both femur and tibia and around the metaphysis of the right distal tibia. Periosteal reaction can be caused by benign or malignant lesions and infection. We cannot find any other cause of periosteal reaction in our case after thorough evaluations. Periosteal reaction in SLE might be associated with inflammatory vascular changes. This is the first report of periosteal reaction in SLE after the 1990s description and the first report in Korea. PMID- 17920324 TI - Influence of RANKL inhibition on immune system in the treatment of bone diseases. AB - Receptor activator of nuclear factor-kappaB ligand (RANKL) is essential for osteoclast differentiation, activation and survival. Recently, denosumab, a human monoclonal antibody to RANKL, has been tested in the treatment of osteoporosis. But the interactions between RANKL, receptor activator of nuclear factor-kappaB (RANK) and osteoprotegerin (OPG) are also involved in immune regulations in addition to bone metabolism. So, blocking RANKL could interplay with immune cells and have systemic effects. Here, we review biological data regarding potential consequences of RANKL blocking on the immune system. PMID- 17920326 TI - "Trigger finger at the wrist" due to anomalous flexor digitorum superficialis muscle belly within the carpal tunnel. AB - "Triggering of the fingers at the wrist" is a relatively unusual condition. It consists in a triggering at the wrist produced by finger motion. Its etiology and presentation may vary. This condition should be clearly differentiated from the other clinical entity called "trigger wrist", occurring on wrist movement. In the present article, we report the case of an anomalous flexor digitorum superficialis muscle belly, arising from the right ring finger at the carpal tunnel, in a 47-year old female patient, causing triggering of the right ring finger at the wrist and a carpal tunnel syndrome. Surgical excision of the muscle mass and carpal tunnel release relieved patient's symptoms and has led to the disappearance of the triggering phenomenon. To date, few cases of trigger finger at the wrist have been reported in the literature. These reported cases were reviewed. The clinical entity of "true trigger wrist" and its etiology are also discussed. PMID- 17920325 TI - [Avulsion of flexor digitorum profundus combined with articular fracture of distal phalanx: a case report]. AB - Avulsion fracture of flexor digitorum profundus is uncomun and usualy related to sports injurys. IV type of Jersey finger is very rare and only few cases have been reported so far. A minimal traumatic form is reported in 34 years old patient; surgery which included open reduction and pullout fixation followed by physiotherapy, led to restoration of a full range of motion of the involved digit. PMID- 17920327 TI - [Radiation induced breast sarcoma: a case report]. AB - Radiation-induced breast sarcoma is a late complication of radiation treatment. We report a case of an undifferentiated sarcoma occurring 8 years after breast conserving treatment, which required mastectomy taking pectoralis major. PMID- 17920328 TI - RPE transplantation and its role in retinal disease. AB - Retinal pigment epithelial (RPE) transplantation aims to restore the subretinal anatomy and re-establish the critical interaction between the RPE and the photoreceptor, which is fundamental to sight. The field has developed over the past 20 years with advances coming from a large body of animal work and more recently a considerable number of human trials. Enormous progress has been made with the potential for at least partial restoration of visual function in both animal and human clinical work. Diseases that have been treated with RPE transplantation demonstrating partial reversal of vision loss include primary RPE dystrophies such as the merTK dystrophy in the Royal College of Surgeons (RCS) rat and in humans, photoreceptor dystrophies as well as complex retinal diseases such as atrophic and neovascular age-related macular degeneration (AMD). Unfortunately, in the human trials the visual recovery has been limited at best and full visual recovery has not been demonstrated. Autologous full-thickness transplants have been used most commonly and effectively in human disease but the search for a cell source to replace autologous RPE such as embryonic stem cells, marrow-derived stem cells, umbilical cord-derived cells as well as immortalised cell lines continues. The combination of cell transplantation with other modalities of treatment such as gene transfer remains an exciting future prospect. RPE transplantation has already been shown to be capable of restoring the subretinal anatomy and improving photoreceptor function in a variety of retinal diseases. In the near future, refinements of current techniques are likely to allow RPE transplantation to enter the mainstream of retinal therapy at a time when the treatment of previously blinding retinal diseases is finally becoming a reality. PMID- 17920329 TI - Negative charged threonine 95 of c-Jun is essential for c-Jun N-terminal kinase dependent phosphorylation of threonine 91/93 and stress-induced c-Jun biological activity. AB - Activation of c-Jun, a major component of the AP-1 transcription factor, represents a paradigm for transcriptional response to stress. Transactivation of c-Jun is regulated by Jun-N-terminal kinases (JNKs) through phosphorylation at serine 63 and 73 (S63/S73), as well as at threonine 91 and 93 (T91/T93). How these two groups of phosphoacceptor sites respond to different grades of genotoxic stress and whether DNA-damage pathways influence the extent of their JNK-dependent phosphorylations remain to be elucidated. Here, we show that following a short exposure to the DNA-damaging compound etoposide, c-Jun phosphorylation is restricted to S63/S73. In contrast, JNK-dependent phosphorylation of T91/T93 requires continuous exposure to the drug and is impaired by caffeine treatment or alanine substitution of the adjacent threonine 95 (T95). Conversely, c-Jun mutations switching the T95/Q96 site into a canonical site for mitogen activated protein kinase (MAPK) phosphorylation (T95/P96) rescues T91/T93 phosphorylation in presence of caffeine, suggesting that a preceding phosphorylation at T95 exposes T91/T93 to JNK-dependent phosphorylation. Moreover, we show that alanine substitution at T95 impairs c-Jun transactivation and c-Jun-mediated cell death, indicating that negatively charged T95 is a general constraint for c-Jun activation. Hence, our study suggests that c-Jun may sense the strength of genotoxic stress through DNA-damage dependent phosphorylation of T95, which in turn augments c-Jun transactivation by JNKs. PMID- 17920331 TI - Tracking down the elusive early endosome. AB - Despite significant progress in understanding protein trafficking and compartmentation in plants, the identification and protein compartmentalization for organelles that belong to both the secretory and endocytic pathways have been difficult because protein trafficking has generally been studied separately in these two pathways. However, recent data indicate that the trans-Golgi network serves as an early endosome merging the secretory and endocytic pathways in plant cells. Here, we discuss the proteins identified as markers for post-Golgi compartments in these two pathways and propose that the trans-Golgi network is a pivotal organelle with multiple sorting domains for post-Golgi protein trafficking in plant cells. PMID- 17920330 TI - Sequential activation of JAKs, STATs and xanthine dehydrogenase/oxidase by hypoxia in lung microvascular endothelial cells. AB - Xanthine dehydrogenase/oxidase (XDH/XO) is associated with various pathological conditions related to the endothelial injury. However, the molecular mechanism underlying the activation of XDH/XO by hypoxia remains largely unknown. In this report, we determined whether the Janus kinases (JAKs) and signal transducers and activators of transcription (STATs) signaling pathway is involved in hypoxia induced activation of XDH/XO in primary cultures of lung microvascular endothelial cells (LMVEC). We found that hypoxia significantly increased interleukin 6 (IL6) production in a time-dependent manner in LMVEC. Hypoxia also markedly augmented phosphorylation/activation of JAKs (JAK1, JAK2 and JAK3) and the JAK downstream effectors STATs (STAT3 and STAT5). Hypoxia-induced activation of STAT3 was blocked by IL6 antibodies, the JAK inhibitor AG490 and the suppressor of cytokine signaling 3 (SOCS3), implying that hypoxia-promoted IL6 secretion activates the JAK/STAT pathway in LMVEC. Phosphorylation and DNA binding activity of STAT3 were also inhibited by the p38 MAPK inhibitor SB203580 and the phosphatidylinositol 3-kinase inhibitor LY294002, suggesting that multiple signaling pathways involved in STAT activation by hypoxia. Importantly, hypoxia promoted XDH/XO activation in LMVEC, which was markedly reversed by inhibiting the JAK-STAT pathway using IL6 antibodies, AG490 and SOCS3. These data demonstrated that JAKs, STATs and XDH/XO were sequentially activated by hypoxia. These data provide the first evidence indicating that the JAK-STAT pathway is involved in hypoxia-mediated XDH/XO activation in LMVEC. PMID- 17920332 TI - Most human metapneumovirus and human respiratory syncytial virus in infant nasal secretions is cell free. AB - BACKGROUND: Nasopharyngeal secretions aspirated from infants with bronchiolitis (NPA) are a valuable resource for the study of virus dynamics and local inflammatory responses, however samples are small and difficult to manipulate. OBJECTIVES: To improve yield of NPA from infants. To establish if removal of the cellular component of NPA affects quantification of human metapneumovirus (hMPV) or human respiratory syncytial virus (hRSV) genome. STUDY DESIGN: Weight of NPA collected into traps from 30 infants was compared with that collected in trap plus catheter and washed through with saline from another 30 infants. hMPV (n=33) and hRSV (n=30) genome was measured by reverse-transcribed real-time polymerase chain reaction (RT-RT-PCR) in paired whole and cell-free NPA collected by the improved method. RESULTS: The improved method of NPA collection gave near two fold greater weight (p = 0.002) of NPA (mean = 0.52 g (S.D. = 0.30 g)) than the traditional method (0.32 g (S.D. 0.19)). There was strong agreement and no significant difference between viral load measured in whole and cell-free fractions of NPA for both viruses (samples (n), correlation coefficient (cc) and significance (p)); hMPV (n=33, cc=0.938, p<0.001) and hRSV (n=30, cc=0.977 and p<0.001). CONCLUSIONS: The majority of hRSV and hMPV in nasal secretions is not associated with cells. Removal of the cellular component of NPA does not interfere with quantification of hRSV and hMPV. PMID- 17920333 TI - Fine needle aspiration biopsy and flow cytometry in the diagnosis of lymphoma. AB - Fine needle aspiration biopsy (FNAB) is emerging as a technique of potential value in the diagnosis of benign and malignant lesions in areas such as the breast and thyroid gland. Its place in distinguishing reactive from neoplastic lymphoid proliferations, when compared to the established practice of excision biopsy and histopathology, continues to undergo evaluation. Morphology alone discriminates poorly between atypical or lymphoproliferative disorders as seen in the presence of Epstein-Barr or human immunodeficiency virus. Furthermore the polymorphic populations of follicular lymphoma may mimic reactive changes. In addition previous classifications of these tumours using working formulation or Kiel classification relied heavily on architecture, which is a feature not reflected in cytology smears. The World Health Organisation approach includes clinical features, immunophenotyping and cytogenetic profiles to define neoplasms of immunohaematopoietic tissues. Flow cytometry on fine needle aspiration biopsy offers additional advantages in being rapid and objective in quantitatively as well as qualitatively documenting cell surface characteristics. All patients referred for this procedure to Tygerberg Academic Hospital with suspected nodal or extranodal lymphoma between January 2002 and December 2004 were analysed. In each case flow cytometry and cytomorphology were correlated with histopathology on tissue biopsy, bone marrow examination and clinical follow-up for confirmation of diagnosis. Results of the 124 cases were tabulated and statistically processed. Eighty-one met the inclusion criteria, thirteen (16.1%) were not malignant, two (2.5%) were falsely negative, two (2.5%) were equivocal needing histology and in the remaining sixty-four (79%) diagnosis was achieved. SUMMARY: Fine needle aspiration coupled with flow cytometry can reliably distinguish between nodal and extranodal neoplastic B-cell population. It is concluded that appropriate use, in a collaborative multidisciplinary setting, may eliminate the need for surgical procedures in many cases. CONCLUSION: These advances are not widely recognised and this is particularly true in South Africa. Accordingly, such an approach has been prospectively evaluated in the Western Cape showing that the combination of ready availability and diagnostic accuracy, after an initial learning curve, allow accurate characterisation of haematologic malignancies so that excision biopsy may be reserved for specific further studies to provide data not available from this less invasive procedure. PMID- 17920334 TI - Sequential imputation for missing values. AB - As missing values are often encountered in gene expression data, many imputation methods have been developed to substitute these unknown values with estimated values. Despite the presence of many imputation methods, these available techniques have some disadvantages. Some imputation techniques constrain the imputation of missing values to a limited set of genes, whereas other imputation methods optimise a more global criterion whereby the computation time of the method becomes infeasible. Others might be fast but inaccurate. Therefore in this paper a new, fast and accurate estimation procedure, called SEQimpute, is proposed. By introducing the idea of minimisation of a statistical distance rather than a Euclidean distance the method is intrinsically different from the thus far existing imputation methods. Moreover, this newly proposed method can be easily embedded in a multiple imputation technique which is better suited to highlight the uncertainties about the missing value estimates. A comparative study is performed to assess the estimation of the missing values by different imputation approaches. The proposed imputation method is shown to outperform some of the existing imputation methods in terms of accuracy and computation speed. PMID- 17920335 TI - Acquired accessory left atrium. AB - The patient was a 61-year-old woman brought to our hospital due to dyspnea and lower extremity swelling in shock state three weeks after mitral valve replacement due to rheumatic mitral regurgitation. Cardiothoracic examination revealed bilateral diffuse rales, metallic first heart sound and a loud holo systolic murmur in apex with radiation to axilla. Transesophageal echocardiography revealed a large echo lucent area representing a separate chamber in the lateral part of left atrium which communicates with left ventricle through a relatively large orifice far from normal functioning prosthesis. In operation left atrial dissection was confirmed but the patient did not wean from cardiopulmonary bypass and died on the operating table. PMID- 17920336 TI - Vanadium and cadmium in vivo effects in teleost cardiac muscle: metal accumulation and oxidative stress markers. AB - Several biological studies associate vanadium and cadmium with the production of reactive oxygen species (ROS), leading to lipid peroxidation and antioxidant enzymes alterations. The present study aims to analyse and compare the oxidative stress responses induced by an acute intravenous exposure (1 and 7 days) to a sub lethal concentration (5 mM) of two vanadium solutions, containing different vanadate n-oligomers (n=1-5 or n=10), and a cadmium solution on the cardiac muscle of the marine teleost Halobatrachus didactylus (Lusitanian toadfish). It was observed that vanadium is mainly accumulated in mitochondria (1.33+/-0.26 microM), primarily when this element was administrated as decameric vanadate, than when administrated as metavanadate (432+/-294 nM), while the highest content of cadmium was found in cytosol (365+/-231 nM). Indeed, decavanadate solution promotes stronger increases in mitochondrial antioxidant enzymes activities (catalase: +120%; superoxide dismutase: +140%) than metavanadate solution. On contrary, cadmium increases cytosolic catalase (+111%) and glutathione peroxidases (+50%) activities. It is also observed that vanadate oligomers induce in vitro prooxidant effects in toadfish heart, with stronger effects induced by metavanadate solution. In summary, vanadate and cadmium are differently accumulated in blood and cardiac subcellular fractions and induced different responses in enzymatic antioxidant defence mechanisms. In the present study, it is described for the first time the effects of equal doses of two different metals intravenously injected in the same fish species and upon the same exposure period allowing to understand the mechanisms of vanadate and cadmium toxicity in fish cardiac muscle. PMID- 17920337 TI - A collaborative medical case authoring environment based on the UMLS. AB - In this paper, we present a novel collaborative authoring tool that was designed to allow medical teachers to formalize and visualize their knowledge for medical intelligent tutoring systems. Our goal is to increase the efficiency and effectiveness in creating the domain model representing the problem solution- often referred to as the bottleneck in developing intelligent tutoring systems. We incorporate the Unified Medical Language System (UMLS) knowledge base to assist the authors in creating the problem solution collaboratively via a videoconferencing platform. The system consists of a shared workspace gathering information visualization and tools necessary for collaborative problem-solving tasks. We found that the authoring tool can be used to effectively elicit the knowledge structure of the domain model. This was achieved in hours compared to months for the conventional paper-based approach. PMID- 17920338 TI - Effect of orlistat on fecal fat, fecal biliary acids, and colonic cell proliferation in obese subjects. AB - BACKGROUND & AIMS: Orlistat is a weight management agent that selectively inhibits gastrointestinal lipase activity. Because of orlistat's mode of action, increased fecal fat is presented to the colonic mucosa, and fecal bile acid and free fatty acid composition may be altered during treatment. Our aim was to assess the effect of treatment of obese subjects with orlistat 120 mg 3 times a day for 6 weeks on fecal lipid and bile acid parameters and colonic mucosal cell proliferation. METHODS: Twenty-four obese (body mass index, 30-40 kg/m2) but otherwise healthy male and female subjects were enrolled in a single-center, randomized, double-blind, placebo-controlled, parallel-group study. Participants were hospitalized during days 1-3 and 33-42 of treatment and were treated as outpatients for the remaining days. RESULTS: Treatment with orlistat for 6 weeks resulted in significantly greater increases in fecal weight, total fecal fat, and fecal free fatty acids than placebo. Total fecal bile acid amounts decreased slightly with orlistat, and increased significantly with placebo treatment (P < .05 between-group difference). Orlistat did not alter colonic cell proliferation as assessed by the 3 proliferative indices (5-bromo-2-deoxyuridine, whole crypt mitotic count, and proliferating cell nuclear antigen). CONCLUSIONS: Biochemical changes in fecal composition related to the pharmacodynamic mode of action of orlistat are not accompanied by altered colonic cell proliferation, a putative biomarker of colon cancer risk. PMID- 17920339 TI - Mouse erythrocyte tropomodulin in the brain reported by lacZ knocked-in downstream from the E1 promoter. AB - Erythrocyte tropomodulin (E-Tmod, Tmod1) is a tropomyosin-binding protein that caps the slow-growing end of actin filaments. In erythrocytes, it may favor the formation of short actin protofilaments needed for elastic cell deformation. Previously we created a knockout mouse model in which lacZ was knocked-in downstream of the E1 promoter to report the expression of full length E-Tmod. Here we utilize E-Tmod(+/lacZ) mice to study E-Tmod expression patterns in the CNS. X-gal staining and in situ hybridization of adults revealed its restricted expression in the olfactory bulb, hippocampus, cerebral cortex, basal ganglia, nuclei of brain stem and cerebellum. In neonates, signals in the cortex and caudate putamen increased from days 15 to 40. Immunohistochemistry also revealed that signals for beta-galactosidase coincided with that of NeuN, a post-mitotic nuclear marker for neurons, but not that for GFAP+ astrocytes or APC+ oligodendrocytes, suggesting E-Tmod/lacZ-positive cells in the CNS were neurons. Large neurons, e.g., mitral cells in olfactory bulb and mossy cells in hilus of the dentate gyrus are among those that expressed very high levels of E-Tmod in the CNS. PMID- 17920340 TI - WITHDRAWN: Dietary Factors, Hormesis and Health. AB - The Publisher regrets that this article is an accidental duplication of an article that has already been published in Ageing Research Reviews, Volume 7, Issue 1, January 2008, Pages 43-48, doi:10.1016/j.arr.2007.08.004. The duplicate article has therefore been withdrawn. PMID- 17920341 TI - DNA polymerase theta (POLQ) can extend from mismatches and from bases opposite a (6-4) photoproduct. AB - DNA polymerase theta (pol theta) is a nuclear A-family DNA polymerase encoded by the POLQ gene in vertebrate cells. The biochemical properties of pol theta and of Polq-defective mice have suggested that pol theta participates in DNA damage tolerance. For example, pol theta was previously found to be proficient not only in incorporation of a nucleotide opposite a thymine glycol or an abasic site, but also extends a polynucleotide chain efficiently from the base opposite the lesion. We carried out experiments to determine whether this ability to extend from non-standard termini is a more general property of the enzyme. Pol theta extended relatively efficiently from matched termini as well as termini with A:G, A:T and A:C mismatches, with less descrimination than a well-studied A-family DNA polymerase, exonuclease-free pol I from E. coli. Although pol theta was unable to, by itself, bypass a cyclobutane pyrimidine dimer or a (6-4) photoproduct, it could perform some extension from primers with bases placed across from these lesions. When pol theta was combined with DNA polymerase iota, an enzyme that can insert a base opposite a UV-induced (6-4) photoproduct, complete bypass of a (6 4) photoproduct was possible. These data show that in addition to its ability to insert nucleotides opposite some DNA lesions, pol theta is proficient at extension of unpaired termini. These results show the potential of pol theta to act as an extender after incorporation of nucleotides by other DNA polymerases, and aid in understanding the role of pol theta in somatic mutagenesis and genome instability. PMID- 17920342 TI - Purification and identification of antiviral components from Laggera pterodonta by high-speed counter-current chromatography. AB - Although Laggera pterodonta, a folk medicine has been widely used for several centuries as an antiviral agent, there have been no studies to identify its antiviral components. A bioassay-guided phytochemical examination of L. pterodonta disclosed that its aqueous extract, which was made up of three dicaffeoylquinic acids showed significant inhibitory activity of virus replication. Then a simple and efficient preparative high-speed counter-current chromatography (HSCCC) method was successfully established by using ethyl acetate n-butanol-water (3:2:5, v/v) as the two-phase solvent system to isolate and purify three bioactive dicaffeoylquinic acids, 3,5-O-dicaffeoylquinic acid, 3,4-O dicaffeoylquinic acid and 4,5-O-dicaffeoylquinic acid. The flow rate was 1.5 ml/min and revolution speed was 800 rpm. The isolation was done in less than 6h, and 34.6 mg of 3,5-O-dicaffeoylquinic acid, 29.4 mg of 3,4-O-dicaffeoylquinic acid and 27.1mg of 4,5-O-dicaffeoylquinic acid was yielded from 600 mg of the crude sample in one-step separation with the purity of 98.3%, 96.7% and 96.2%, respectively, as determined by high-performance liquid chromatography (HPLC). The structures of these three bioactive dicaffeoylquinic acids were identified by ultraviolet (UV), electrospray ionization mass spectrometry (ESI-MS), proton nuclear magnetic resonance ((1)H NMR) and carbon-13 nuclear magnetic resonance (13C NMR). In the antiviral experiment, three dicaffeoylquinic acids all showed significant inhibitory activity against herpes simplex virus-1 (HSV-1), herpes simplex virus-2 (HSV-2) and influenza viruses A (IVA) in vitro with high selectivity indexes. However, among the three compounds, 3,5-O-dicaffeoylquinic acid and 4,5-O-dicaffeoylquinic acid were the more active than 3,4-O dicaffeoylquinic acid against HSV-1, HSV-2 and IVA. This study demonstrated a direct link between the antiviral activity of L. pterodonta and the three dicaffeoylquinic acids. PMID- 17920343 TI - Sperm activation: role of reactive oxygen species and kinases. AB - Reactive oxygen species (ROS), such as the superoxide anion (O(2)(-*)), hydrogen peroxide (H(2)O(2)) and nitric oxide (NO*), when generated at low and controlled levels, act as second messengers. ROS regulate sperm capacitation, which is the complex series of changes allowing spermatozoa to bind to the zona pellucida surrounding the oocyte, induce the acrosome reaction (exocytotic event by which proteolytic enzymes are released) and fertilize the oocyte. Capacitating spermatozoa produce controlled amounts of ROS that regulate downstream events: first, the increase in cAMP, protein kinase A (PKA) activation and phosphorylation of PKA substrates (arginine-X-X-serine/threonine motif; 15-30 min); second, the phosphorylation of MEK (extracellular signal regulated kinase [ERK] kinase)-like proteins (30-60 min) and then that of the threonine-glutamate tyrosine motif (>1 h); finally, the late tyrosine phosphorylation of fibrous sheath proteins (>2 h). Although all these events are ROS-dependent, the regulation by various kinases, protein kinase C, PKA, protein tyrosine kinases, the ERK pathway, etc. is different. ROS also regulate the acquisition of hyperactivated motility and the acrosome reaction by spermatozoa. ROS action is probably mediated via the sulfhydryl/disulfide pair on sperm proteins. Redundancy, cross talk, and multiple systems acting in parallel point to an array of safeguards assuring the timely function of spermatozoa. PMID- 17920344 TI - Fabrication of bioactive glass-ceramic foams mimicking human bone portions for regenerative medicine. AB - A technique for the preparation of bioglass foams for bone tissue engineering is presented. The process is based on the in situ foaming of a bioglass-loaded polyurethane foam as the intermediate step for obtaining a bioglass porous monolith, starting from sol-gel synthesized bioglass powders. The obtained foams were characterized using X-ray diffraction analysis, Fourier transform infrared spectroscopy, and field emission scanning electron microscopy observations. The material was assessed by soaking samples in simulated body fluid and observing apatite layer formation. Diagnostic imaging taken from human patients was used to reconstruct a human bone portion, which was used to mould a tailored scaffold fabricated using the in situ foaming technique. The results confirmed that the obtained bioactive materials prepared with three-dimensional processing are promising for applications in reconstructive surgery tailored to each single patient. PMID- 17920345 TI - A reliable method for the placement and securement of tissue expander remote injection port. PMID- 17920346 TI - Shwachman-Diamond syndrome is associated with low-turnover osteoporosis. AB - INTRODUCTION: Shwachman-Diamond syndrome (SDS) is an autosomal recessive disorder characterized by exocrine pancreatic insufficiency and bone marrow dysfunction. These result in malabsorption and hematological abnormalities. A skeletal dysplasia is also an integral feature of SDS. The present study assessed prevalence and determinants of osteopenia and osteoporosis in patients with SDS and disease-causing mutations in the SBDS gene. MATERIALS AND METHODS: Eleven patients (8 males) aged from 5 to 37 years (median 16.7 years) with a genetically confirmed diagnosis of SDS were assessed for fracture history, bone mineral content (BMC), lean tissue mass (LTM) and bone mineral density (BMD) (Hologic Discovery A), osteoporotic vertebral changes, and for blood biochemistry and hematological parameters. Iliac crest bone biopsies were obtained from four patients for histology and histomorphometry. RESULTS: The main findings were: (1) markedly reduced BMD Z-scores at the lumbar spine (median -2.1, range -4.4 to 0.8), proximal femur (median -1.3, range -2.2 to -0.7) and, whole body (median 1.0, range -2.8 to +0.6), and reduced Z-scores for height-adjusted BMC/LTM ratio (median -0.9, range -3.6 to +1.1); (2) vertebral compression fractures in three patients; and (3) blood biochemistry suggestive of mild vitamin D and vitamin K deficiency. Bone biopsies in four patients showed significant low-turnover osteoporosis with reduced trabecular bone volume, low numbers of osteoclasts and osteoblasts, and reduced amount of osteoid. CONCLUSIONS: The results suggest that in addition to the skeletal dysplasia, SDS is associated with a more generalized bone disease characterized by low bone mass, low bone turnover and by vertebral fragility fractures. Osteoporosis may result from a primary defect in bone metabolism, and could be related to the bone marrow dysfunction and neutropenia. PMID- 17920348 TI - (18)F-labeled positron emission tomographic radiopharmaceuticals in oncology: an overview of radiochemistry and mechanisms of tumor localization. AB - Molecular imaging is the visualization, characterization, and measurement of biological processes at the molecular and cellular levels in a living system. At present, positron emission tomography/computed tomography (PET/CT) is one the most rapidly growing areas of medical imaging, with many applications in the clinical management of patients with cancer. Although [(18)F]fluorodeoxyglucose (FDG)-PET/CT imaging provides high specificity and sensitivity in several kinds of cancer and has many applications, it is important to recognize that FDG is not a "specific" radiotracer for imaging malignant disease. Highly "tumor-specific" and "tumor cell signal-specific" PET radiopharmaceuticals are essential to meet the growing demand of radioisotope-based molecular imaging technology. In the last 15 years, many alternative PET tracers have been proposed and evaluated in preclinical and clinical studies to characterize the tumor biology more appropriately. The potential clinical utility of several (18)F-labeled radiotracers (eg, fluoride, FDOPA, FLT, FMISO, FES, and FCH) is being reviewed by several investigators in this issue. An overview of design and development of (18)F-labeled PET radiopharmaceuticals, radiochemistry, and mechanism(s) of tumor cell uptake and localization of radiotracers are presented here. The approval of clinical indications for FDG-PET in the year 2000 by the Food and Drug Administration, based on a review of literature, was a major breakthrough to the rapid incorporation of PET into nuclear medicine practice, particularly in oncology. Approval of a radiopharmaceutical typically involves submission of a "New Drug Application" by a manufacturer or a company clearly documenting 2 major aspects of the drug: (1) manufacturing of PET drug using current good manufacturing practices and (2) the safety and effectiveness of a drug with specific indications. The potential routine clinical utility of (18)F-labeled PET radiopharmaceuticals depends also on regulatory compliance in addition to documentation of potential safety and efficacy by various investigators. PMID- 17920349 TI - Cancer imaging with fluorine-18-labeled choline derivatives. AB - The choline transporter and choline kinase enzyme frequently are overexpressed in malignancy. Therefore, positron-emitter-labeled compounds derived from choline have the potential to serve as oncologic probes for positron emission tomography. The fluorine-18 ((18)F)-labeled choline derivative fluorocholine (FCH) in particular has demonstrated potential utility for imaging of a variety of neoplasms, including those of the breast, prostate, liver, and brain. The pharmacokinetics of FCH and other choline tracers allow for whole-body imaging within minutes of injection while still achieving high tumor-to-background contrast in most organs, including the brain. These features, along with the possibility of imaging malignancies that have proved elusive with the use of (18)F-fluorodeoxyglucose positron emission tomography support further clinical investigations of (18)F-labeled choline tracers. PMID- 17920350 TI - FLT: measuring tumor cell proliferation in vivo with positron emission tomography and 3'-deoxy-3'-[18F]fluorothymidine. AB - Positron emission tomography (PET) using the radiotracer 3'-deoxy-3' [(18)F]fluorothymidine (FLT) can image cellular proliferation in human cancers in vivo. FLT uptake has been shown to correlate with pathology-based proliferation measurements, including the Ki-67 score, in a variety of human cancers. Unlike pathology-based measurements, imaging-based methods, including FLT-PET, are noninvasive, easily repeatable, and less prone to sampling errors. FLT-PET may therefore be a useful tool for assessing tumor aggressiveness, predicting outcome, planning therapy, or monitoring response to treatment. Three recent clinical studies have reported that FLT-PET can accurately predict response very early after the initiation of chemotherapy. Especially with the advent of cytostatic chemotherapy agents, methods of biologically assessing a tumor's response will take on increasing importance, since changes in tumor size will not always be expected. To date, most studies of FLT-PET have focused on validating it as a means of quantifying cellular proliferation and testing its ability to accurately stage cancer. In some settings, FLT-PET has shown greater specificity for cancer than (18)F-fluorodeoxyglucose (FDG)-PET, which can show false-positive uptake in areas of infection or inflammation. However, because of FLT's lower overall uptake and higher background activity in liver and bone marrow, FLT-PET should not be considered a potential replacement for staging by FLT-PET. Instead, FLT-PET should be considered a powerful addition to FDG-PET, providing additional diagnostic specificity and important biological information that could be useful in predicting prognosis, planning treatment, and monitoring response. PMID- 17920351 TI - 3,4-dihydroxy-6-[18f]-fluoro-L-phenylalanine positron emission tomography in patients with central motor disorders and in evaluation of brain and other tumors. AB - The use of 3,4-dihydroxy-6-[(18)F]-fluoro-L-phenylalanine ((18)F-FDOPA) with positron emission tomography initially centered on studying central motor disorders and evaluating patients with Parkinsonian symptoms, based on its uptake into presynaptic dopaminergic terminals in the putamen and caudate nuclei of the brain. The roles of this tracer have since expanded to include monitoring disease progression, potentially contributing to drug development, and even questioning the current gold standard for making the diagnosis of Parkinson's disease. As with some other amino acids, (18)F-FDOPA has also been effective for visualizing brain tumors, either at time of diagnosis or when monitoring for recurrence, with high sensitivity and overall accuracy. (18)F-FDOPA may be especially useful for imaging patients with low-grade gliomas, as well in the evaluation of patients with neuroendocrine tumors such as carcinoid and pheochromocytoma, in which its role as a precursor for amine neurotransmitter/neurohormones serves as a basis for its differential uptake. PMID- 17920352 TI - Hypoxia positron emission tomography imaging with 18f-fluoromisonidazole. AB - The importance of hypoxia in disease pathogenesis and prognosis is gathering increasing clinical significance and having a greater impact on patient management and outcome. Previous efforts to evaluate hypoxia have included the invasive assessment of hypoxia with immunohistologic and histographic oxygen probes. The emergence of new radiotracers has allowed noninvasive assessment of hypoxia, with the most extensively investigated and validated positron emission tomography radiotracer of hypoxia to date being (18)F-fluoromisonodazole ((18)F FMISO). This review discusses the relevance and biology of hypoxia in cells and organ systems, and reviews the laboratory and clinical applications of (18)F FMISO in oncology and noncancer disease states. PMID- 17920353 TI - 18F-Fluoride positron emission tomography and positron emission tomography/computed tomography. AB - (18)F-Fluoride is a positron-emitting bone-seeking agent, the uptake of which reflects blood flow and remodeling of bone. Assessment of (18)F-fluoride kinetics using quantitative positron emission tomography (PET) methods allows the regional characterization of lesions of metabolic bone diseases and the monitoring of their response to therapy. It also enables the assessment of bone viability and discrimination of uneventful and impaired healing processes of fractures, bone grafts and osteonecrosis. Taking advantage of the favorable pharmacokinetic properties of the tracer combined with the high performance of PET technology, static (18)F-fluoride PET is a highly sensitive imaging modality for detection of benign and malignant osseous abnormalities. Although (18)F-fluoride uptake mechanism corresponds to osteoblastic activity, it is also sensitive for detection of lytic and early marrow-based metastases, by identifying their accompanying reactive osteoblastic changes, even when minimal. The instant fusion of increased (18)F-fluoride uptake with morphological data of computed tomography (CT) using hybrid PET/CT systems improves the specificity of (18)F-fluoride PET in cancer patients by accurately differentiating between benign and malignant sites of uptake. The results of a few recent publications suggest that (18)F fluoride PET/CT is a valuable modality in the diagnosis of pathological osseous conditions in patients also referred for nononcologic indications. (18)F-fluoride PET and PET/CT are, however, not widely used in clinical practice. The limited availability of (18)F-fluoride and of PET and PET/CT systems is a major factor. At present, there are not enough data on the cost-effectiveness of (18)F-fluoride PET/CT. However, it has been stated by some experts that (18)F-fluoride PET/CT is expected to replace (99m)Tc-MDP bone scintigraphy in the future. PMID- 17920354 TI - 18F-Fluoroestradiol. AB - Estrogen receptor (ER) expression is an important determinant of breast cancer behavior and is critical for response to endocrine therapies such as tamoxifen and aromatase inhibitors. In current practice, ER expression is determined by assay of biopsy material. In more advanced disease, tissue assay may present practical difficulties and be associated with significant sampling error. This and other considerations motivated the development of ER imaging agents for positron emission tomography (PET), of which the most successful has been (18)F 16alpha-17beta-fluoroestradiol (FES). In this review, we highlight aspects of ER biology and the importance of the ER in breast cancer therapy; review the structure and synthesis of FES; describe its kinetics and safety/dosimetry data; and highlight validation studies. Also discussed are early results in patients using FES-PET to localize ER-expressing tumors and associated data pointing toward its accuracy as a predictive assay for breast cancer endocrine therapy. Finally, early data for tumors and sites other than breast cancer are mentioned. Preliminary data strongly point toward potential clinical utility for FES-PET, motivating further validation and future clinical trials with prospective endpoints tested under appropriate regulatory oversight. PMID- 17920355 TI - Comparison of outcomes using sirolimus-eluting stenting in diabetic versus nondiabetic patients with comparison of insulin versus non-insulin therapy in the diabetic patients. AB - The effect of insulin therapy on adverse cardiovascular outcomes in diabetic patients has been debated and a reduced benefit in clinical restenosis outcomes after sirolimus stenting has been reported among diabetic patients requiring insulin therapy. We analyzed 297 diabetic patients receiving sirolimus-eluting stents, including 115 (39%) on insulin therapy, and compared outcomes with 541 nondiabetic patients treated consecutively during the same interval. The rates of target lesion revascularization (9.5% vs 3.5%, p = 0.003) and cardiac death or myocardial infarction (MI, 7.1% vs 3.1%, p = 0.012) were significantly higher for diabetic patients. Insulin treatment was independently associated with increased risk for target lesion revascularization (odds ratio [OR] 2.48, 95% confidence interval [CI] 1.22 to 5.00) and cardiac death or MI (hazard ratio [HR] 2.85, 95% CI 1.41 to 5.77), whereas the adjusted risk for diabetic patients not treated with insulin was not significantly different from patients without diabetes for target lesion revascularization (OR 1.32, 95% CI 0.66 to 2.62) or cardiac death or MI (HR 1.04, 95% CI 0.50 to 2.17). In conclusion, diabetes mellitus is associated with increased risk for target lesion revascularization and cardiac death or MI after receiving sirolimus-eluting stenting, and is significantly exaggerated by the requirement for insulin therapy. PMID- 17920356 TI - Differences in utilization of drug-eluting stents by race and payer. AB - Numerous disparities in access to health care by race and gender have been identified in the literature. This study examines differences in the use of drug eluting stents (DES) versus bare-metal stents (BMS) by race, payer, and income level. Data from New York State's Percutaneous Coronary Intervention Reporting System from July 2003 to December 2004 were used to examine use of DES (20,165 patients) relative to BMS (4,547 patients) by race, payer, and annual income level, controlling for a variety of patient and hospital characteristics. African Americans were found to be less likely to receive DES than other races between July 2003 and March 2004 (adjusted odds ratio [OR] 0.56, 95% confidence interval [CI] 0.50 to 0.65) and between April 2004 and December 2004 (adjusted OR 0.74, 95% CI 0.61 to 0.90). These disparities were reduced (respective adjusted ORs 0.67, 95% CI 0.58 to 0.77 and 0.81, 95% CI 0.66 to 0.91) when controlling for admitting hospital and hospital volume, but were still significant. Medicaid/self pay patients, and patients living in zip codes with median annual incomes between $20,000 and $30,000 were also less likely to receive DES in the first time period (adjusted respective ORs 0.80, 95% CI 0.68 to 0.93) and 0.85, 95% CI 0.75 to 0.96). In conclusion, African-Americans and low income groups receive DES less frequently than their counterparts compared with BMS. This is related to the hospitals where they are admitted, but not entirely. PMID- 17920357 TI - Meta-analysis appraising high clopidogrel loading in patients undergoing percutaneous coronary intervention. AB - Combined antiplatelet treatment with aspirin and clopidogrel is pivotal to minimize periprocedural adverse events in patients who undergo percutaneous coronary intervention. However, there is debate on the best clopidogrel loading dose. The investigators performed a systematic review and meta-analysis of the optimal clopidogrel loading dose. Pertinent trials comparing high (>300 mg) and standard (300 mg) clopidogrel loading doses in patients scheduled for catheterization and/or percutaneous coronary intervention were systematically searched in BioMedCentral, CENTRAL, Google Scholar, and PubMed (December 2006). The primary end point was the 1-month rate of death or myocardial infarction. Secondary end points included other ischemic and bleeding adverse effects. Peto odds ratios were computed. A total of 10 studies (7 randomized, 3 nonrandomized) were included, enrolling 1,567 patients (712 loaded with 300 mg, 11 with 450 mg, 790 with 600 mg, and 54 with 900 mg). Overall, a high loading dose proved significantly superior to a standard loading dose in preventing cardiac death or nonfatal myocardial infarction (odds ratio 0.54, 95% confidence interval 0.32 to 0.90, p = 0.02), without any statistically significant increase in major or minor bleedings (p = 0.55 and p = 0.98, respectively). Sensitivity analysis restricted to randomized trials confirmed the superiority of a high loading dose regimen (p = 0.0031). Meta-regression disclosed a significant interaction between event rate and the benefits of high loading doses (p = 0.005), suggesting that the greater the underlying risk, the greater the favorable impact of a high loading dose. In conclusion, a high clopidogrel loading dose (>300 mg) significantly reduces early ischemic events in patients scheduled for percutaneous coronary intervention. PMID- 17920358 TI - High yellow color intensity by angioscopy with quantitative colorimetry to identify high-risk features in culprit lesions of patients with acute coronary syndromes. AB - High yellow color intensity (HYCI) regions of atherosclerotic plaque, determined by angioscopy with quantitative colorimetry, are associated with lipid cores underneath thin fibrous caps in ex vivo tissue samples. To determine whether HYCI regions of coronary plaque are associated with disruption or thrombus in living patients, quantitative colorimetry was applied to angioscopy, and the color of culprit lesions was measured in patients with acute coronary syndromes. In 46 patients with acute coronary syndromes (acute myocardial infarction, n = 14; unstable angina pectoris [UAP] with culprit thrombus, n = 16; and UAP without culprit thrombus, n = 16), the recorded angioscopic images of culprit lesions were analyzed using a quantitative colorimetric method based on the L*a*b* color space applied to angioscopy (positive b* = yellow color intensity). HYCI was defined as b* value >23. Plaque disruption was significantly more prevalent in 19 of 24 HYCI regions (79%) than in 9 of 22 non-HYCI regions (41%) (p = 0.007). Culprit HYCI regions were prevalent in patients with myocardial infarction (11 of 14 [79%]), followed by those with UAP with thrombus (9 of 16 [56%]) and UAP without thrombus (4 of 16 [25%]) (p = 0.01 for trend), and were significantly more prevalent in 66% of patients with myocardial infarction and UAP with thrombus compared with 25% of those with UAP without thrombus (p = 0.007). In conclusion, HYCI regions of coronary plaque may be indicative of high-risk lesions vulnerable to thrombosis. Coronary angioscopy with quantitative colorimetry could be used to study the association between high-risk coronary lesions and future cardiovascular events. PMID- 17920359 TI - Effect of intracoronary thrombectomy on 30-day mortality in patients with acute myocardial infarction. AB - Insertion of intracoronary thrombectomy (ICT) devices, as a precedent to percutaneous coronary intervention (PCI), theoretically could have a beneficial effect on the outcome in patients with acute myocardial infarction. To examine whether ICT was associated with a lower 30-day mortality rate in patients with acute myocardial infarction, we studied 3,913 patients who underwent PCI within 24 hours after onset. A total of 990 patients (25.3%) were treated with ICT before PCI. The 30-day mortality rate was lower in the patients receiving ICT than in those without (3.7% vs 6.2%, p = 0.004), but this beneficial effect disappeared after adjustment for baseline characteristics (hazard ratio [HR] 0.658, p = 0.166). We also divided the patients into tertiles according to the Thrombolysis In Myocardial Infarction (TIMI) risk score. After adjustment for baseline characteristics, ICT was associated with a lower 30-day mortality rate in patients from the highest TIMI risk score tertile (HR 0.407, p = 0.029), but not in patients from the lower 2 tertiles. ICT was also an independent predictor of a lower 30-day mortality risk in patients aged > or =70 years (HR 0.239, p = 0.007), patients with diabetes mellitus (HR 0.275, p = 0.039), and those with stent implantation (HR 0.437, p = 0.034). In conclusion, in selected patients with high TIMI risk scores, an age > or =70 years, diabetes mellitus, or stenting, ICT is associated with a lower 30-day mortality rate. PMID- 17920360 TI - Long-term outcomes of patients with mitral regurgitation undergoing percutaneous coronary intervention. AB - The most appropriate treatment for patients with ischemic mitral regurgitation (MR) remains controversial. MR has prognostic importance in patients after myocardial infarction and those undergoing coronary artery bypass surgery, but the long-term outcomes after percutaneous coronary intervention (PCI) are less well defined. We evaluated patients who underwent PCI and had assessment of MR by left ventriculography and/or echocardiography in the year 2000. We determined effects of MR on 30-day and 5-year survival. The cohort included 711 patients (67% men) with an average age of 64.5 +/- 12.4 years. MR severity was divided into 3 strata: none (n = 420, 59%), mild (n = 209, 29%), and moderate to severe (n = 82, 12%). Patients with more severe MR differed from patients with mild or no MR in that they were older (p <0.001), more frequently women (p <0.001), and more likely to have a coronary artery bypass graft (p <0.001), myocardial infarction (p <0.001), and lower ejection fraction (p <0.001). Decreased survival rates were associated with increasing MR severity (none vs mild vs moderate to severe) at 30 days (100%, 98.7%, and 96.6%, respectively; p <0.0025) and 5 years (97%, 83.3%, and 57.5%; p <0.0001). MR was an important independent predictor of survival (hazard ratio 1.57, p <0.0009). In conclusion, patients with ischemic MR undergoing PCI have significantly decreased survival rates at 5 years, and severity of MR is an independent predictor of survival. PMID- 17920361 TI - Comparison of age, body mass index, and frequency of systemic hypertension and diabetes mellitus in patients having coronary angioplasty in 1996 versus in 2006. AB - The data submitted to the New York State Department of Health Coronary Angioplasty Reporting System Database on coronary angioplasties performed at Westchester Medical Center/New York Medical College from 1996 to 2005 were analyzed. Compared with 1996, during 2005, more coronary angioplasties were performed (1,624 vs 1,122), and the patients were older (mean age 64.5 vs 61.0 years, p <0.001), weighed more (84.2 vs 82.0 kg, p <0.001), had a higher mean body mass index (28.8 vs 28.3 kg/m(2), p <0.001), and had a higher prevalence of diabetes mellitus (27% vs 17%, p <0.001). The prevalence of systemic hypertension was significantly higher in 2005 (76%) than in 1996 (54%) (p <0.001). In conclusion, in 2005 compared with 1996, patients who underwent coronary angioplasty at Westchester Medical Center/New York Medical College were older, had higher body mass indexes, and had higher prevalences of diabetes mellitus and systemic hypertension. PMID- 17920362 TI - Impact of delay in door-to-needle time on mortality in patients with ST-segment elevation myocardial infarction. AB - Fibrinolytic therapy is the most common reperfusion strategy for patients with ST segment elevation myocardial infarction (STEMI), particularly in smaller centers. Previous studies evaluated the relation between time to treatment and outcomes when few patients were treated within 30 minutes of hospital arrival and many did not receive modern adjunctive medications. To quantify the impact of a delay in door-to-needle time on mortality in a recent and representative cohort of patients with STEMI, a cohort of 62,470 patients with STEMI treated using fibrinolytic therapy at 973 hospitals that participated in the National Registry of Myocardial Infarction from 1999 to 2002 was analyzed. Hierarchical models were used to evaluate the independent effect of door-to-needle time on in-hospital mortality. In-hospital mortality was lower with shorter door-to-needle times (2.9% for < or =30 minutes, 4.1% for 31 to 45 minutes, and 6.2% for >45 minutes; p <0.001 for trend). Compared with those experiencing door-to-needle times < or =30 minutes, adjusted odd ratios (ORs) of dying were 1.17 (95% confidence interval [CI] 1.04 to 1.31) and 1.37 (95% CI 1.23 to 1.52; p for trend <0.001) for patients with door-to-needle times of 31 to 45 and >45 minutes, respectively. This relation was particularly pronounced in those presenting within 1 hour of symptom onset to presentation time (OR 1.25, 95% CI 1.01 to 1.54; OR 1.54, 95% CI 1.27 to 1.87, respectively; p for trend <0.001). In conclusion, timely administration of fibrinolytic therapy continues to significantly impact on mortality in the modern era, particularly in patients presenting early after symptom onset. PMID- 17920363 TI - Peri-stent reference segment plaque burden is associated with disease progression in saphenous vein grafts (a serial intravascular ultrasound assessment). AB - We are aware of no studies of peri-stent disease progression or luminal compromise in saphenous vein graft (SVG) lesions. We used serial intravascular ultrasound (IVUS) to assess disease progression in peri-stent saphenous vein bypass graft reference segments. We studied 37 peri-stent SVG reference segments in 21 patients; 16 were proximal and 21 were distal to the stent. The same anatomic image slice was analyzed after the intervention and at follow-up; this site was 3.68 +/- 2.22 mm from the stent edge. Graft age was 10.1 +/- 5.4 years, and mean follow-up duration was 13 months (range 3 to 61). Overall, change in SVG area, change in lumen area, and change in plaque burden correlated with postintervention plaque burden (r = 0.448, p = 0.005; r = -0.584, p <0.001; and r = 0.507, p = 0.001, respectively). For the proximal edge, change in lumen area correlated with change in plaque area (r = -0.951, p <0.001), but not with change in SVG area (r = -0.337, p = 0.201). For the distal edge, change in lumen area correlated more strongly with change in plaque area (r = -0.982, p <0.001) than with change in SVG area (r = -0.624, p = 0.003). When peri-stent reference segments were divided into 2 groups according to postintervention plaque burden (>50% [n = 20] vs <50% [n = 17]), there was a greater decrease in lumen area ( 1.12 +/- 0.81 vs -0.33 +/- 0.26 mm(2), p <0.001) and greater increases in SVG area (0.26 +/- 0.29 vs 0.09 +/- 0.09 mm(2), p = 0.027), plaque area (1.37 +/- 0.96 vs 0.42 +/- 0.30 mm(2), p <0.001), and plaque burden (8.2 +/- 5.6% vs. 2.8 +/- 1.6%, p <0.001) in segments with a plaque burden >50%. In conclusion, peri stent reference segment SVG disease progression and lumen loss were more significant in segments with a greater postintervention plaque burden after implantation of a bare metal stent or drug-eluting stent. PMID- 17920364 TI - Comparison between dobutamine echocardiography and single-photon emission computed tomography for interpretive reproducibility. AB - Interpretive variability of dobutamine stress echocardiography (DSE) and stress single-photon emission computed tomography (SPECT) has been previously investigated. However, no study has directly compared the variability of these techniques in the same patient population. We directly compared the interpretive reproducibility of DSE and stress SPECT in patients undergoing both types of pharmacologic stress imaging. Before discharge, simultaneous DSE and SPECT was performed in 56 patients early after a first acute myocardial infarction. Intra- and interobserver concordances were evaluated by exact agreement and kappa statistic. Intraclass coefficient of correlation was used to assess intra- and interobserver reproducibilities of segmental score analysis. Intraobserver agreement percentages in the identification of patients with ischemia were 98% for SPECT and 91% for DSE (p = NS) and kappa values were excellent (>0.80) for both techniques. Interobserver agreement was higher (p <0.01) for SPECT (96%) than for DSE (79%). Similarly, kappa value was excellent for SPECT (0.92) and only moderate for DSE (0.56). Finally, the intraclass coefficients of correlation for intra- and interobserver reproducibilities were higher for SPECT (0.98 and 0.97, respectively) than for DSE (0.80 and 0.71, respectively; p <0.001 for both). In conclusion, after uncomplicated acute myocardial infarction, stress SPECT imaging has a better interpretive reproducibility than DSE. PMID- 17920365 TI - Comparison of effectiveness of rosuvastatin versus atorvastatin on the achievement of combined C-reactive protein (<2 mg/L) and low-density lipoprotein cholesterol (< 70 mg/dl) targets in patients with type 2 diabetes mellitus (from the ANDROMEDA study). AB - Decreasing C-reactive protein (CRP) in addition to decreasing low-density lipoprotein (LDL) cholesterol may further decrease coronary heart disease risk. The effects of rosuvastatin compared with atorvastatin in achieving a combined target of LDL cholesterol <70 mg/dl and CRP <2 mg/L in 509 patients with type 2 diabetes mellitus was evaluated. CRP decreased significantly versus baseline in both treatment groups. Significantly more patients treated with rosuvastatin achieved the combined end point of LDL cholesterol <70 mg/dl and CRP <2 mg/L compared with atorvastatin by the end of the study period (58% vs 37%; p <0.001 vs atorvastatin). In conclusion, CRP was effectively decreased in patients with type 2 diabetes receiving rosuvastatin or atorvastatin, whereas rosuvastatin decreased LDL cholesterol significantly more than atorvastatin. PMID- 17920366 TI - Differential effects of intravenous magnesium on atrioventricular node conduction in supraventricular tachycardia. AB - Evidence from noninvasive studies suggests magnesium has a differential effect on atrioventricular nodal (AVN) pathways. To further explore the electrophysiologic effects of intravenous magnesium sulfate (MgSO(4)) on supraventricular tachycardia, with particular reference to AVN conduction pathways, we studied 23 patients with supraventricular tachycardia at the time of electrophysiologic study. Tachycardia cycle length; AH, HV, and VA intervals; anterograde and retrograde Wenckebach thresholds; slow and fast pathway effective refractory periods (ERPs); accessory pathway ERP; right atrial and ventricular ERPs; blood pressure; and serum magnesium were evaluated before and after administration of MgSO(4) during sustained tachycardia. AVN reentry was induced in 14 patients and atrioventricular reentry was induced in 9; 1 of the latter had dual AVN physiology with tachycardia using the slow pathway. Serum magnesium level increased from 0.88 +/- 0.11 to 1.79 +/- 0.14 mmol/L (p <0.0001). Magnesium increased tachycardia cycle length to a greater extent in those with dual AVN physiology than those without: 340 +/- 54 to 370 +/- 57 ms versus 347 +/- 29 to 350 +/- 30 ms (p = 0.01). This was associated with greater increase in AH interval in those with dual AVN physiology than in those without: 241 +/- 59 to 270 +/- 60 ms versus 144 +/- 16 to 140 +/- 20 ms (p = 0.003). Presence of dual AVN physiology was more frequently associated with reversion to sinus rhythm: 5 of 15 versus 0 of 8 (p = 0.06). MgSO(4) did not alter other measured parameters. In conclusion, magnesium increases tachycardia cycle length and AH interval in patients with dual AVN physiology through a dominant effect on the slow AVN pathway. PMID- 17920367 TI - A meta-analysis of 94,492 patients with hypertension treated with beta blockers to determine the risk of new-onset diabetes mellitus. AB - Beta blockers used for the treatment of hypertension may be associated with increased risk for new-onset diabetes mellitus (DM). A search of Medline, PubMed, and EMBASE was conducted for randomized controlled trials of patients taking beta blockers as first-line therapy for hypertension with data on new-onset DM and follow-up for > or =1 year. Twelve studies evaluating 94,492 patients fulfilled the inclusion criteria. Beta-blocker therapy resulted in a 22% increased risk for new-onset DM (relative risk 1.22, 95% confidence interval [CI] 1.12 to 1.33) compared with nondiuretic antihypertensive agents. A higher baseline fasting glucose level (odds ratio [OR] 1.01, 95% CI 1.00 to 1.02, p = 0.004) and greater systolic (OR 1.05, 95% CI 1.05 to 1.08, p = 0.001) and diastolic (OR 1.06, 95% CI 1.01 to 1.10, p = 0.011) blood pressure differences between the 2 treatment modalities were significant univariate predictors of new-onset DM. Multivariate meta-regression analysis showed that a higher baseline body mass index (OR 1.17, 95% CI 1.01 to 1.33, p = 0.034) was a significant predictor of new-onset DM. The risk for DM was greater with atenolol, in the elderly, and in studies in which beta blockers were less efficacious antihypertensive agents and increased exponentially with increased duration on beta blockers. For the secondary end points, beta blockers resulted in a 15% increased risk for stroke, with no benefit for the end point of death or myocardial infarction. In conclusion, beta blockers are associated with an increased risk for new-onset DM, with no benefit for the end point of death or myocardial infarction and with a 15% increased risk for stroke compared with other agents. This risk was greater in patients with higher baseline body mass indexes and higher baseline fasting glucose levels and in studies in which beta blockers were less efficacious antihypertensive agents compared with other treatments. PMID- 17920368 TI - Usefulness of tissue Doppler velocity and strain dyssynchrony for predicting left ventricular reverse remodeling response after cardiac resynchronization therapy. AB - The assessment of systolic dyssynchrony by echocardiography is useful in predicting a favorable response to cardiac resynchronization therapy (CRT). Tissue Doppler velocity and tissue Doppler longitudinal strain have been suggested for this purpose. This study compared parameters of systolic dyssynchrony derived from these 2 imaging modalities for their predictive values of CRT response. Two hundred fifty-six patients from 3 different centers who received CRT were followed for 6 +/- 3 months. Parameters of systolic dyssynchrony based on tissue Doppler velocity and strain imaging were assessed for the prediction of left ventricular (LV) reverse remodeling (reduction of LV end-systolic volume > or =15%). These included time to peak systolic velocity (or peak strain) of 12 LV segments to calculate the SD (Ts-SD or Tepsilon-SD), maximal difference in delay (Ts-Diff or Tepsilon-Diff), and opposite wall delay (Ts-OW or Tepsilon-OW). The septal-to-lateral delay (Ts-Sep-Lat or Tepsilon-Sep Lat) was also measured. LV reverse remodeling, defined as improvement in end systolic volume > or =15%, was observed in 141 patients (55%). All 4 tissue velocity parameters predicted LV reverse remodeling, and the areas under the receiver-operating characteristic curves were 0.86, 0.85, 0.84, and 0.79 for Ts SD, Ts-Diff, Ts-OW, and Ts-Sep-Lat, respectively (all p <0.001). The cut-off values derived from receiver-operating characteristic curve analysis were 33 ms for Ts-SD, 100 ms for Ts-Diff, 90 ms for Ts-OW, and 60 ms for Ts-Sep-Lat, and their sensitivities were 93%, 92%, 81%, and 70%, with specificities of 78%, 68%, 80%, and 76%, respectively. In contrast, none of the longitudinal strain parameters predicted LV reverse remodeling. The areas under the receiver operating characteristic curves ranged from 0.49 to 0.53 (all p = NS). The same conclusions were obtained in subgroup analyses of QRS duration (120 to 150 vs >150 ms) and ischemic or nonischemic cause of heart failure. In conclusion, parameters of tissue Doppler longitudinal velocity, but not longitudinal strain, predicted LV reverse remodeling after CRT. PMID- 17920369 TI - Comparison of Afro-Caribbean patients presenting in heart failure with normal versus poor left ventricular systolic function. AB - Data suggest that heart failure (HF) in Afro-Caribbean patients may be more often associated with preserved left ventricular (LV) systolic function, LV hypertrophy, and probable LV diastolic dysfunction than in other populations. Echocardiographic results on all patients referred for HF in a contemporary Afro Caribbean population were reviewed, comparing findings in patients with and without preserved LV systolic function with. Echocardiographic findings included left atrial dimension, LV systolic and diastolic dimensions, ventricular septal and posterior wall thicknesses, right ventricular dimension, valve abnormality, or pericardial effusion. LV shortening fraction and ejection fraction were calculated. Age, gender, and presence of atrial fibrillation were recorded. Results from patients with preserved LV systolic function (LV shortening fraction >0.27) were compared with those with poor LV systolic function. There were 505 patients with HF with adequate studies; mean age +/- SD was 64 +/- 15 years, 46% were men, 17% had atrial fibrillation, and 285 of 505 (57%) had preserved LV systolic function. Those with preserved LV systolic function were no different in age (64 +/- 15 vs 64 +/- 14 years, p = 0.98) but were less likely to be men (40% vs 54%, p <0.01). They were less likely to have a dilated left atrium (61% vs 81%, p <0.001) or increased LV diastolic dimension (8% vs 63%, p <0.001). They were more likely to have increased ventricular septal or posterior wall hypertrophy (84% vs 66%, p <0.001) or other abnormal findings, including an abnormal valve, right ventricular enlargement, increased septal to posterior wall thickness ratio, or pericardial effusion (25% vs 6%, p <0.001). The presence of atrial fibrillation was no different (14% vs 20%, p = 0.10). In conclusion, most Afro-Caribbean patients with HF have preserved LV systolic function with high rates of LV hypertrophy, septal hypertrophy, and other echocardiographic abnormalities. PMID- 17920370 TI - Prognostic value of exercise tolerance testing in asymptomatic chronic nonischemic mitral regurgitation. AB - In many heart diseases, exercise tolerance testing (ETT) has useful functional correlates and/or prognostic value. However, its predictive value in mitral regurgitation (MR) is undefined. To determine whether ETT descriptors predict death or indications for mitral valve surgery in patients with MR, we prospectively followed, for 7 +/- 3 end-point-free years, a cohort of 38 patients with chronic severe nonischemic MR who underwent modified Bruce ETT; all lacked surgical indications at study entry. Their baseline exercise descriptors were also compared with those from 46 patients with severe MR who, at entry, already had reached surgical indications. End points during follow-up in the cohort included sudden death (n = 1), heart failure symptoms (n = 2), atrial fibrillation (n = 4), left ventricular (LV) ejection fraction <60% (n = 2), LV systolic dimensions > or =45 mm (n = 12) and >40 mm (n = 11), LV ejection fraction <60% plus LV systolic dimensions > or =45 mm (n = 3), and heart failure plus LV systolic dimensions > or =45 mm plus LV ejection fraction <60% (n = 1). In univariate analysis, exercise duration (p = 0.004), chronotropic response (p = 0.007), percent predicted peak heart rate (p = 0.01), and heart rate recovery (p <0.02) predicted events; in multivariate analysis, only exercise duration was predictive (p <0.02). Average annual event risk was fivefold lower (4.62%) with an exercise duration > or =15 versus <15 minutes (average annual risk 23.48%, p = 0.004). Relative risks in patients with and without exercise-inducible ST-segment depression were comparable (< or =1.3, p = NS) whether defined at entry and/or during follow-up. Exercise duration, but not prevalence of exercise-inducible ST segment depression, was lower (p <0.001) in patients with surgical indications at entry versus initially end-point-free patients. In conclusion, in asymptomatic patients with chronic severe nonischemic MR and no objective criteria for operation, progression to surgical indications generally is rapid. However, those with excellent exercise tolerance have a relatively benign course. Exercise inducible ST-segment depression has no prognostic value in this population. PMID- 17920371 TI - Epidemiologic features of infective endocarditis in Taiwanese adults involving native valves. AB - Little is known about the incidence and clinical outcomes of infective endocarditis (IE) involving native valves in Asian countries. This nationwide study investigated epidemiologic features and in-hospital mortality associated with IE in adults (age > or =18 years) based on Taiwan's National Health Insurance database from 1997 through 2002. Of 7,240 enrolled patients with IE involving native valves, the mean age was 53 +/- 19 years and 70% were men. The mean annual crude incidence was 7.6 per 100,000 inhabitants. The incidence was significantly higher in men than in women (10.4 vs 4.6 per 100,000; p <0.001). The incidence of IE increased steadily with age, ranging from 3.8 per 100,000 persons in patients <30 years of age to 33 per 100,000 persons in patients > or =80 years of age (p <0.001). Staphylococcal (32%) and streptococcal species (61%) were the most common causative pathogens. The mean in-hospital mortality rate was 18%. Multivariate analysis showed that male gender, older age (> or =50 years), diabetes mellitus, heart failure, neurologic complications, renal insufficiency, respiratory failure, shock, and Staphylococcus species as the causative microorganism were independent predictors of in-hospital mortality. In conclusion, this Taiwanese study revealed a high incidence of IE in men and elderly subjects. The in-hospital mortality rate remained high. Patients with IE who also developed shock and respiratory failure were the most likely to have a poor outcome. PMID- 17920372 TI - Valve structure and survival in sexagenarians having aortic valve replacement for aortic stenosis (+/-aortic regurgitation) with versus without coronary artery bypass grafting at a single US medical center (1993 to 2005). AB - The purpose of this study was to determine the effect of simultaneous coronary artery bypass grafting (CABG) and the influence of valve structure on both early and late survival in sexagenarians having aortic valve replacement (AVR) for aortic stenosis (AS) (with or without aortic regurgitation). We analyzed survival and valve structure in 289 sexagenarians having AVR for AS from 1993 through 2005 at Baylor University Medical Center, including 147 (51%) with and 142 (49%) without simultaneous CABG. Of the 282 patients with information available, 13 (4.6%) died within 30 days of operation and 1 additional patient, from 31 to 60 days after operation (5.0% 60-day mortality). Sixty-day mortality was similar (6% and 4%) in the groups with and without simultaneous CABG. A total of 66 patients (23%) died from >60 days up to 13 years postoperatively. The unadjusted survival analysis showed that late survival was not affected by gender (male versus female), aortic valve structure (unicuspid, bicuspid, and quadricuspid versus tricuspid) or preoperative severity of the AS (transvalvular peak pressure gradient >50 mm Hg versus < or =50 mm Hg), or by performance of CABG. The aortic valve was congenitally unicuspid in 10 patients (3%), congenitally bicuspid in 170 (59%), 3-cuspid in 107 (37%), congenitally quadricuspid in 1 patient, and the valve structure was indeterminate in 1 patient. In conclusion, gender, valve structure, preoperative severity of the AS, or performance of simultaneous CABG did not effect unadjusted survival in sexagenarians undergoing AVR for AS. PMID- 17920374 TI - Acute effect of esmolol intravenously on coronary microcirculation in patients with idiopathic dilated cardiomyopathy. AB - Although coronary flow reserve (CFR) impairment was correlated with the prognosis of patients with idiopathic dilated cardiomyopathy (IDC) and microvascular ischemia was implicated in the progress of the disease, little is known about the effect of the established therapy with beta blockers on coronary microcirculation. The purpose of this study was to assess the effect of beta(1) blockade on coronary blood flow and CFR in patients with IDC. Fourteen patients with IDC and 10 control subjects underwent time-averaged peak coronary flow velocity (APCFV) measurements (centimeters per second) in the proximal left anterior descending coronary artery at baseline and at maximal hyperemia before and after beta(1) blockade with intravenous esmolol. CFR was defined as APCFV at maximal hyperemia/APCFV at baseline. Although there were no significant differences in APCFV at baseline between patients with IDC and controls, patients with IDC had significantly lower APCFV at maximal hyperemia than controls (54.2 +/- 12.0 vs 75.1 +/- 18.6, p <0.05) and decreased CFR (2.39 +/- 0.38 vs 3.50 +/- 0.54, respectively, p <0.05). After beta(1) blockade, a significant decrease in APCFV at baseline (19.5 +/- 3.7 vs 22.9 +/- 5.0, p <0.05) and enhancement of APCFV at maximal hyperemia (59.5 +/- 13.3 vs 54.2 +/- 12.0, p <0.05) were observed in patients with IDC, but not in control subjects, leading to significant improvement in CFR (3.06 +/- 0.40 vs 2.39 +/- 0.38, p <0.05). In conclusion, patients with IDC had alterations in coronary blood flow and decreased CFR that improved after beta(1) blockade. These alterations in microvascular function, which are partially reversed by beta blockade, may be 1 of the underlying mechanisms that contribute to the improved prognosis of patients with IDC under such therapy. PMID- 17920373 TI - Right ventricular involvement in hypertrophic cardiomyopathy. AB - The aim of this study was to assess, using cardiovascular magnetic resonance (CMR), whether morphologic right ventricular (RV) abnormalities are present in patients with hypertrophic cardiomyopathy (HC). Left ventricular hypertrophy has been considered the predominant phenotypic expression of HC. Whether structural abnormalities of the right ventricle are also present in HC is unknown. CMR provides complete coverage of both ventricles with high spatial resolution. CMR was applied to study RV morphology in HC. CMR was performed on 46 subjects with HC (mean age 39 +/- 16 years; 70% men) free of pulmonary hypertension and 22 healthy subjects (mean age 44 +/- 16 years; 50% men). Mass, wall thickness, chamber volume, the ejection fraction, and fibrosis were assessed for both ventricles. Maximum RV wall thickness was increased in patients with HC compared with referent controls (7 +/- 2 vs 5 +/- 1 mm, p <0.001), including 15 (33%) with maximum wall thicknesses > or =8 mm (> or =2 SDs higher than the mean for controls) and 4 (9%) with extreme hypertrophy (> or =10 mm). RV hypertrophy was predominantly a diffuse process involving the entire or a significant proportion of the RV wall in most patients (n = 8 [53%]). The RV wall mass index was also increased in patients with HC (28 +/- 9 vs 22 +/- 4 g, p <0.001). A significant correlation was found between maximum RV and left ventricular wall thickness (R(2) = 0.4, p <0.001) and between RV and left ventricular mass (R(2) = 0.4, p <0.001). Only 1 (2%) patient with HC had evidence of RV wall fibrosis. In conclusion, morphologic RV abnormalities are present in a substantial proportion of patients with HC. PMID- 17920375 TI - Ventricular systolic reserve in asymptomatic children previously treated with low doses of anthracyclines. AB - Doppler echocardiography has been used for the diagnosis of anthracycline-induced cardiotoxicity. However, few data are available that include asymptomatic children previously treated with a low cumulative dose of this drug and therefore have a low risk of cardiac dysfunction. The aim of this study was to evaluate after-exercise cardiac function in asymptomatic children previously treated with a low cumulative dose of anthracycline and no clinical or laboratory evidence of cardiotoxicity. Doppler echocardiography was performed before and immediately after physical exercise in 29 children aged 5 to 17 years (anthracycline [ADRIA] group). All had finished cancer treatment with anthracycline derivatives for > or =1 year (cumulative dose 100 mg/m(2)). Results were compared with those from age- and gender-matched healthy children (control group; n = 26) using the Mann Whitney rank test. Exercise-induced cardiac function changes within groups were analyzed using Wilcoxon's signed-rank test. Exercise induced significant increases in left ventricular systolic function indexes in both groups. However, the ADRIA group had significantly lower changes in left ventricular ejection fraction (ADRIA group 0.71 +/- 0.02 vs 0.80 +/- 0.04 and control group 0.71 +/- 0.02 vs 0.89 +/- 0.05, p = 0.0017) and end-systolic stress-volume index (ADRIA group 4.59 +/- 0.69 vs 6.4 +/- 2.0 g.cm(-2)/ml.m(-2) and control group 5.49 +/- 0.98 vs 11.54 +/- 2.86 g.cm(-2)/ml.m(-2); p <0.0001), indicating decreased functional systolic reserve. In conclusion, asymptomatic children previously treated with low cumulative doses of anthracycline had decreased functional systolic reserve evidenced by exercise, suggesting a nonclinically manifested cardiotoxicity. PMID- 17920376 TI - Prevalence of hepatitis C infection in adult patients who underwent congenital heart surgery prior to screening in 1992. AB - Before the routine screening of donor blood for hepatitis C virus (HCV) began in 1992, patients who received blood transfusions were at increased risk for HCV infection. Patients with congenital heart disease requiring surgery were at particular risk because of the large volume of blood products required. It was hypothesized that patients who had congenital heart surgery before 1992 would have a high prevalence of HCV infection and would benefit from routine screening as part of cardiologic follow-up. HCV screening is performed on all patients in the Adult Congenital Cardiac Program at the investigators' center. Of 198 patients who underwent heart surgery before 1992, 17 (8.6%) had positive HCV antibody results, and 8 (4.0%) had positive HCV ribonucleic acid results. Only 1 of these patients had elevated serum transaminase levels at the time of screening. In conclusion, the data from this study demonstrated an approximately fivefold increased prevalence of HCV infection in adults with congenital heart disease compared with the aged-matched general population. Serum transaminase levels are not a valid screening method. The long-term significance of HCV in this population is not known. Routine HCV screening for all patients who underwent cardiac surgery before 1992 is recommended. PMID- 17920377 TI - Utility of B-type natriuretic peptide in predicting medium-term mortality in patients undergoing major non-cardiac surgery. AB - This study was conducted to assess the ability of preoperative B-type natriuretic peptide levels to predict medium-term mortality in patients who undergo major noncardiac surgery. During a median of 654 days of follow-up, 33 patients from a total cohort of 204 patients (16%) died, 17 from cardiovascular causes. The optimal cutoff in this cohort, determined using a receiver-operating characteristic curve, was >35 pg . ml(-1). This was associated with a 3.5-fold increase in the hazard of death (p = 0.001) and a 6.9-fold increase in the hazard of cardiovascular mortality (p = 0.003). In conclusion, these findings extend recent work demonstrating that B-type natriuretic peptide levels obtained before major noncardiac surgery can be used to predict perioperative morbidity and indicate that they also forecast medium-term mortality, particularly cardiovascular death. PMID- 17920378 TI - Clinical and anatomical characteristics of subtle-discrete dissection of the ascending aorta. AB - Subtle or discrete (class 3 in the classification of the European Society of Cardiology) dissection is the most neglected variant of aortic dissection. This study was conducted to define the clinical manifestations, diagnostic findings, and outcomes of subtle or discrete dissection involving the ascending aorta. The clinical and surgical records, preoperative studies, and outcomes of 109 consecutive patients with ascending aortic dissection observed from 1995 to 2005 were reviewed. Eight patients (7.3%) had discrete dissection. Five patients presented with acute anterior chest pain, 2 with abdominal pain, and 4 with syncope. The mean diameter of the ascending aorta was 44 +/- 8.8 mm. The intimal tears were located in all patients on the posterior aspect of the ascending aorta 1 to 40 mm above the left coronary ostium; its length varied from 2.8 to 12.3 mm. Preoperative aortography, magnetic resonance imaging, and computed tomography could not identify the discrete intimal tears. Transesophageal echocardiography provided unique diagnostic information on (1) subtle intimal discontinuity, (2) circumscribed intramural hematoma, and (3) discrete pericardial fluid around the dissected aorta. Six patients underwent emergency surgery on the basis of echocardiographic findings, and they were all alive at follow-up. Compared with patients with classic aortic dissection, those with discrete dissection had lower operative mortality (0% vs 26%, p = 0.11), shorter hospital stay (7.2 +/- 2.8 vs 21 +/- 19 days, p = 0.01), and less frequent need for blood transfusions (0% vs 39%, p = 0.02). In conclusion, elevated clinical suspicion and detailed transesophageal echocardiographic examination are important for the early identification of discrete aortic dissection, leading to prompt surgery, shorter hospital stays, and better outcomes. PMID- 17920379 TI - A prescription for improving national health care. PMID- 17920380 TI - Morphologic validation of reperfused hemorrhagic myocardial infarction by cardiovascular magnetic resonance. AB - The purposes of this study were to assess the ex vivo cardiovascular magnetic resonance (CMR) signals of pathologically proved hemorrhagic myocardial infarction (MI) and to correlate these with in vivo CMR findings. Late gadolinium hypoenhancement within a hyperenhanced area in reperfused acute MI is ascribed to severe microvascular obstruction. The hearts of 2 patients, who died from cardiogenic shock after acute MIs and who had undergone coronary recanalization and in vivo CMR, were examined by T(2) and T(1) late enhancement sequences as well as by gross and histologic investigation. Four corresponding short-axis slices of each cardiac specimen from the base to the left ventricular apex were selected to assess the extent of MI and hemorrhage and were compared with the in vivo T(2) and late enhancement CMR scans. On pathologic examination, the extent of MI was 57 +/- 30% and 44 +/- 24%, and the extent of hemorrhage was 23 +/- 13% and 19 +/- 8% of the left ventricular area, respectively, showing progressive increases from the base to the apex. The low-signal intensity areas observed by ex vivo T(2) CMR strongly correlated with the hemorrhage quantified on histology (R = 0.93, p = 0.0007). Using ex vivo late gadolinium sequences, bright areas surrounded by thin dark rims, consistent with magnetic susceptibility effects, were detected, corresponding with hemorrhage. On in vivo CMR images, low-signal intensity and hyperintense areas with peripheral susceptibility artifacts were observed within the MI core on T(2) and late gadolinium sequences, respectively. In conclusion, in reperfused MI, CMR hypointense T(2) signal and susceptibility effects within the late gadolinium hypoenhanced areas are consistent with interstitial hemorrhage due to irreversible vascular injury, as proved by pathologic study. PMID- 17920381 TI - Aneurysms of sinus of Valsalva. PMID- 17920382 TI - Myopathy, statins, and vitamin D deficiency. PMID- 17920383 TI - Statins and vitamin D. PMID- 17920384 TI - Response to Tobis critique of computed tomographic angiography. PMID- 17920385 TI - Retinopathy as a useful guide for selecting an appropriate coronary revascularization treatment in patients with diabetes mellitus. PMID- 17920386 TI - Optical sensing systems for microfluidic devices: a review. AB - This review deals with the application of optical sensing systems for microfluidic devices. In the "off-chip approach" macro-scale optical infrastructure is coupled, while the "on-chip approach" comprises the integration of micro-optical functions into microfluidic devices. The current progress of the use of both optical sensing approaches in microfluidic devices, as well as its applications is described. In all cases, sensor size and shape profoundly affect the detection limits, due to analyte transport limitation, not to signal transduction limitation. The micro- or nanoscale sensors are limited to picomolar order detection for practical time scales. The review concludes with an assessment of future directions of optical sensing systems for integrated microfluidic devices. PMID- 17920387 TI - Using classification structure pharmacokinetic relationship (SCPR) method to predict drug bioavailability based on grid-search support vector machine. AB - The linear discriminant analysis (LDA) and the grid-search support vector machine (GS-SVM) were used to develop classification structure pharmacokinetic relationship models for predicting drug bioavailability. Bioavailability data for 167 compounds were taken from the literature, and the molecular descriptors were generated from the software CODESSA solely from molecular structures. Five descriptors selected by LDA were used to build the linear and nonlinear models. The obtained results confirmed the discriminative capacity of the calculated descriptors and the relationship with the drug bioavailability. The result of GS SVM (total accuracy of 85.6%) was better than that of LDA (total accuracy of 72.4%), which indicated that the GS-SVM model was more reliable in the recognition of the drug bioavailability. The proposed method was very useful for the selection of new drugs products, and can also be extended in other classification structure pharmacokinetic relationship (CSPR) and classification structure activity relationship (CSAR) investigation. PMID- 17920388 TI - Sensitive determination of iron(III) by gold electrode modified with 2 mercaptosuccinic acid self-assembled monolayer. AB - Preparation and application of gold 2-mercaptosuccinic acid self-assembled monolayer (Au-MSA SAM) electrode for determination of iron(III) in the presence of iron(II) is described by cyclic voltammetry, electrochemical impedance spectroscopy, and Osteryoung square wave voltammetry. The square wave voltammograms showed a sharp peak around positive potentials +0.250 V that was used for construction of the calibration curve. Parameters influencing the method were optimized. A linear range calibration curve from 1.0x10(-10) to 6.0x10(-9) M iron(III) with a detection limit of 3.0x10(-11) M and relative standard deviation (R.S.D.) of 6.5% for n=8 at 1.0x10(-9) M iron(III) was observed in the best conditions. Possible interferences from the coexisting ions were also investigated. The results demonstrated that sensor could be used for determination of iron(III) in the presence of various ions. The validity of the method and applicability of the sensor were successfully tested by determining of iron(III) in natural waters (tap and mineral waters) and in a pharmaceutical sample (Venofer ampoule) without interference from sample matrix. The experimental data are presented and discussed from which the new sensor is characterized. PMID- 17920389 TI - Use of organofunctionalized nanoporous silica gel to improve the lifetime of carbon paste electrode for determination of copper(II) ions. AB - A new functionalized nanoporous silica gel with dipyridyl group (DPNSG) was synthesized. Then, the potentiometric response of the copper(II) ion was investigated at a carbon paste electrode chemically modified with this newly designed functionalized nanoporous silica gel. The electrodes with DPNSG proportions of 15.0% (w/w) demonstrated very stable potentials. Calibration plots with Nernstian slopes for Cu2+ were observed, 28.4 (+/-1.0) mV decade(-1), over a wide linear concentration range (1.0x10(-7) to 1.0x10(-2) M). The electrode exhibited a detection limit of 8.0x10(-8) M. Moreover, the selectivity coefficients measured by the match potential method in acetate buffer, pH 5.5, were investigated. The electrode presented a response time of approximately 50s, high performance, high sensitivity in a wide range of cation activities and good long-term stability (more than 9 months). The method was satisfactory and was used to determine the copper ion concentration in waste waters, contaminated by this metal. PMID- 17920390 TI - Catalytic spectrophotometric determination of iodine in coal by pyrohydrolysis decomposition. AB - A method for the determination of iodine in coal using pyrohydrolysis for sample decomposition was proposed. A pyrohydrolysis apparatus system was constructed, and the procedure was designed to burn and hydrolyse coal steadily and completely. The parameters of pyrohydrolysis were optimized through the orthogonal experimental design. Iodine in the absorption solution was evaluated by the catalytic spectrophotometric method, and the absorbance at 420 nm was measured by a double-beam UV-visible spectrophotometer. The limit of detection and quantification of the proposed method were 0.09 microg g(-1) and 0.29 microg g(-1), respectively. After analysing some Chinese soil reference materials (SRMs), a reasonable agreement was found between the measured values and the certified values. The accuracy of this approach was confirmed by the analysis of eight coals spiked with SRMs with an indexed recovery from 94.97 to 109.56%, whose mean value was 102.58%. Six repeated tests were conducted for eight coal samples, including high sulfur coal and high fluorine coal. A good repeatability was obtained with a relative standard deviation value from 2.88 to 9.52%, averaging 5.87%. With such benefits as simplicity, precision, accuracy and economy, this approach can meet the requirements of the limits of detection and quantification for analysing iodine in coal, and hence it is highly suitable for routine analysis. PMID- 17920391 TI - Calibration transfer strategy to compensate for instrumental drift in portable quadrupole mass spectrometers. AB - We report the use of a calibration transfer strategy to correct for drift in the quantitative sensitivity of a portable quadrupole mass spectrometer (QMS) aimed at process monitoring applications. Gas mixtures of CH4/Ar/C2H6/CO2 were studied with calibration phase measurements made of the pure gases for a univariate analysis and of 40 multi-component mixtures for a multivariate approach. To evaluate calibrations, test set spectra of a CH4/Ar/C2H6/CO2 gas mixture were recorded bi-weekly over a period of 12 months. As part of the strategy a standard of pure argon was measured during both calibration and test phases so that correction factors could be calculated for each measurement day. It was shown that in the absence of a calibration transfer strategy quantifications of test set spectra could be inaccurate by more than an order of magnitude over 12 months. Furthermore, due to the effects of drift in the sensitivity over the 6 days required to record the training set in the calibration phase it was found that the multivariate analysis quantified test spectra less accurately than the univariate analysis. However, by applying the calibration transfer strategy across all measurements (both calibration and test phases) it was shown that the errors in prediction using the multivariate analysis previously seen after 2 weeks were not observed until approximately 12 months later. PMID- 17920393 TI - Fiber optic-linear array detection spectrophotometry in combination with cloud point extraction for simultaneous preconcentration and determination of cobalt and nickel. AB - A new combined method including fiber optic-linear array detection spectrophotometry (FO-LADS) and cloud point extraction (CPE) was developed using a cylindrical micro cell for simultaneous preconcentration and determination of different species. The CPE and FO-LADS methods have good matching conditions for combination because FO-LADS is suitable as a detection technique for the low volume of remained phase obtained after CPE. This combination was carried out using 50 microL cylindrical micro cell and then employed for simultaneous preconcentration and determination of cobalt and nickel. Cloud point extraction method was based on the chromogenic reaction of metal ions and 1-(2-pyridylazo)-2 naphthol (PAN) and then preconcentration of formed complexes using octylphenoxypolyethoxyethanol (Triton X-114). The remained phase after CPE was transferred into cylindrical micro cell and located at the cell holder of FO LADS. The spectra of cobalt and nickel complexes were collected by FO-LADS and processed for ordinary and first derivative spectrophotometry. Optimization of different parameters was evaluated. Under optimum conditions, calibration curves were linear in the range of 0.6-30.0 and 0.1-15.0 microg L(-1) with detection limits of 0.2 and 0.04 microg L(-1) for Co and Ni respectively. The relative standard deviations (R.S.D.s) were lower than 4%. The obtained enhancement factors were 198 and 199 for cobalt and nickel, respectively. The proposed method was compared with the other methods and applied to the analysis of several real and spiked samples. PMID- 17920392 TI - A novel flow-through fluorescence optosensor for the determination of thiabendazole. AB - This paper presents the development of a new flow-injection system combined with solid-surface fluorescence detection for the determination of the widely used fungicide thiabendazole. Nylon powder was probed as a novel solid support for building the optosensor. The method is based on the on-line immobilization of thiabendazole onto nylon in a continuous flow system, followed by the measurement of its native fluorescence. Aqueous samples are directly injected in a water carrier, resulting in a very simple and economical method. The analytical figures of merit obtained using 1500 microL of sample and 75% methanol (v/v) as eluting solution were: linear calibration range from 8 to 120 ng mL(-1) (the lowest value corresponds to the quantitation limit), relative standard deviation, 0.9% (n=5) at a level of 64 ng mL(-1), limit of detection calculated according to 1995 IUPAC recommendations is to 2.8 ng mL(-1), and sampling rate of 14 samples h(-1). The potential interference from other agrochemicals, metal ions and common anions, and the viability of determining thiabendazole in real water samples were also evaluated. PMID- 17920394 TI - A new high-performance liquid chromatography assay for the determination of sesquiterpene lactone 15-deoxygoyazensolide in rat plasma. AB - A simple, rapid and sensitive high-performance liquid chromatography method was developed for the analysis of the sesquiterpene lactone 15-deoxygoyazensolide (LAC15-D) in rat plasma samples. The chromatographic separation was achieved on a LiChrospher RP18 column using methanol:water (50:50, v/v) containing 0.6% acetic acid as mobile phase, at a flow rate of 0.7 mL min(-1). UV detection was carried out at 270 nm. Phenytoin was used as internal standard. Prior to the analysis, the rat plasma samples were submitted to liquid-liquid extraction with dichloromethane. The mean absolute recoveries were 73% with R.S.D. values lower than 3.5. The method was linear over the 6.0-2000 ng mL(-1) concentration range and the quantification limit was 6.0 ng mL(-1). Within-day and between-day assay precision and accuracy were studied at three concentration levels (15, 300 and 480 ng mL(-1)) and were lower than 15%. The validated method was used to measure the plasmatic concentration of LAC15-D in rats that received a single intraperitoneal dose of 30 mg kg(-1). PMID- 17920395 TI - Sorption characteristics and chromatographic separation of gold (I and III) from silver and base metal ions using polyurethane foams. AB - The influence of different parameters on the sorption profiles of trace and ultra traces of gold (I) species from the aqueous cyanide media onto the solid sorbents ion exchange polyurethane foams (IEPUFs) and commercial unloaded polyurethane foams (PUFs) based polyether type has been investigated. The retention of gold (I) species onto the investigated solid sorbents followed a first-order rate equation with an overall rate constant k in the range 2.2-2.8+/-0.2 s(-1). The sorption data of gold (I) followed Freundlich and Langmuir isotherm models. Thus, the a dual-mode of sorption mechanism involving absorption related to "weak base anion exchanger" and an added component for "surface adsorption" seems the most likely proposed dual mechanism for retention profile of gold (I) by the IEPUFs and PUFs solid sorbents. The capacity of the IEPUFs and PUFs towards gold (I) sorption calculated from the sorption isotherms was found to be 11.21+/-1.8 and 5.29+/-0.9 mg g(-1), respectively. The chromatographic separation of the spiked inorganic gold (I) from de ionized water at concentrations 5-15 microg mL(-1) onto the developed IEPUFs and PUFs packed columns at 10 mL min(-1) flow rate was successfully achieved. The retained gold (I) species were then recovered quantitatively from the IEPUFs (98.4+/-2.4%, n=5) and PUFs (95.4+/-3.4%, n=5) packed columns using perchloric acid (60 mL, 1.0 mol L(-1)) as a proper eluting agent. Thiourea (1.0 mol L(-1))-H2SO4 (0.1 mol L(-1)) system was also used as eluting agent for the recovery of gold (I) from IEPUFS (95.4+/-5.4%, n=3) and also PUFs (93.4+/-4.4%, n=3) packed columns. The performance of the IEPUFs and PUFs packed columns in terms of the height equivalent to the theoretical plates (HETP), number of plates (N), and critical and breakthrough capacities towards gold (I) species were evaluated. The developed IEPUFs packed column was applied successfully for complete retention and recovery (98.5+/-2.7) of gold (III) species spiked onto tap- and industrial wastewater samples at <10 microg Au mL( 1) after reduction to gold (I). The IEPUFs packed column was applied satisfactorily for complete retention and recovery (98.5+/-2.7) of total inorganic gold (I) and/or gold (III) species spiked to tap- and industrial wastewater samples at <10 microg mL(-1) gold. Chromatographic separation of gold (I) from silver (I) and base metal ions (Fe, Ni, Cu and Zn) using IEPUFS packed columns was satisfactorily achieved. The proposed method was applied successfully for the pre-concentration and separation from anodic slime and subsequent FAAS determination of analyte with satisfactory results (recoveries >95%, relative standard deviations <4.0%). PMID- 17920396 TI - Simultaneous screening analysis of multiple beta-blockers in urine by gas chromatography-mass spectrometry in selected ion monitoring mode. AB - A rapid screening procedure is described for the simultaneous determination of 13 beta-blockers in urine at the range of 0.010-1.0 microg mL(-1). The procedure involves N-ethoxycarbonyl (EOC) derivatization of beta-blockers in urine sample, followed by extraction and further conversion to trimethylsilyl (TMS) derivatives for the analysis by gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode (GC-SIM-MS). The characteristic fragment ions at m/z 260 and m/z 144, and [M-15]+ ions permitted sensitive and selective detection of most of the beta-blockers in the presence of co-extracted urinary amino alcohols at much higher levels. The whole procedure of EOC/TMS derivatization with subsequent GC-SIM-MS analysis was linear (r > or = 0.9988). The LODs were varied from 0.03 to 2.7 ng mL(-1). The ranges of precision (%relative standard deviation) and accuracy (%relative error) of the overall procedure at two different added amounts (0.02 and 0.5 microg mL(-1)) in urine matrix varied from 1.3 to 9.4 and from -9.6 to 9.7, respectively. The recoveries were measured to be ranged from 90.4 to 109.7%. PMID- 17920397 TI - Traceability of synthetic drugs by position-specific deuterium isotope ratio analysis: the case of Prozac and the fluoxetine generics. AB - Samples of fluoxetine of different origin were submitted to natural abundance 2H NMR spectroscopy. The deuterium content at the various sites of the molecule was found to depend on its synthetic history. Hints on the synthetic procedure can be obtained by comparison with standard compounds, whose synthesis is known. These preliminary results give an idea of the potential of site-specific isotope ratio analysis in the fight against patent infringement and drug counterfeiting. PMID- 17920402 TI - Substandard application of preimplantation genetic screening may interfere with its clinical success. AB - The intent of this study was to evaluate a recent randomized clinical trial evaluating the effect of preimplantation genetic screening (PGS) that reports a negative effect on pregnancy outcome. This article reviews appropriate PGS techniques and how they differ from the trial in question. A closer look at the clinical trial in question reveals significant lack of expertise in biopsy, cell fixation, genetic analysis, and patient selection. At most, this trial demonstrates that in inexperienced hands, PGS can be detrimental. No other conclusions concerning the effect of PGS on pregnancy results can be drawn from the trial. PMID- 17920403 TI - A comparison of letrozole to gonadotropins for ovulation induction, in subjects who failed to conceive with clomiphene citrate. AB - OBJECTIVE: To compare pregnancy rates (PR) for letrozole and gonadotropins in individuals who failed to conceive with clomiphene citrate (CC). DESIGN: Retrospective cohort study. SETTING: University reproductive center. PATIENT(S): Individuals treated with letrozole or gonadotropins who failed to conceive with CC. INTERVENTION(S): Controlled ovarian hyperstimulation (COH), transvaginal ultrasound, ovulation induction, IUI. MAIN OUTCOME MEASURE(S): Pregnancy rates per cycle. RESULT(S): Among patients who failed to conceive with at least three cycles of CC, gonadotropins had a higher PR per cycle than letrozole. Among individuals who failed to conceive with less than three cycles of CC and whose medications were changed because of thin uterine lining or intolerable side effects, average PR per cycle for letrozole and gonadotropin treatments were equivalent. All patients conceived within three stimulation cycles with either gonadotropins or letrozole. CONCLUSION(S): In patients who failed to conceive with CC, gonadotropins have higher PR for ovulation induction than letrozole. However, PR were high enough with letrozole to justify its use in this population of patients. Letrozole and gonadotropins should not be used for more than three cycles without a conception. PMID- 17920404 TI - Stage I ovarian carcinoma: different clinical pathologic patterns. AB - OBJECTIVE: To analyze clinicopathologic patterns of early ovarian carcinoma. DESIGN: Retrospective chart and histopathology review. SETTING: Mount Sinai School of Medicine, New York and the Centre Jean Perrin, Clermont Ferrand, France. PATIENT(S): Seventy-six consecutive cases of Federation Internationale de Gynecologie et d'Obstetrique stage I ovarian carcinoma. INTERVENTION(S): Surgical staging. MAIN OUTCOME MEASURE(S): Symptomatology, pathology, and histology analysis. RESULT(S): Twenty-two cases (29%) were serous papillary carcinomas and 54 were nonserous carcinomas (71%) (40 endometrioid, 10 clear cell, and 4 mixed endometrioid and clear cell carcinomas). Ninety-eight percent of ovarian endometriosis, 95% of endometrial carcinomas, and 83% of endometrial polyps and hyperplasias were associated with nonserous carcinomas. Most patients with serous papillary carcinoma presented with asymptomatic pelvic masses; patients with nonserous carcinomas presented with pelvic pain or abnormal vaginal bleeding with or without pelvic mass. CONCLUSION(S): Over two thirds of stage I ovarian carcinomas were nonserous, and were diagnosed because of associated symptoms: pelvic pain with endometriosis and/or adnexal masses, or vaginal bleeding from endometrial pathology. Serous papillary carcinomas were often asymptomatic and diagnosed during follow-up evaluations in breast cancer patients. Stage I ovarian carcinoma has different clinical and pathologic patterns than advanced ovarian carcinoma. The risk of ovarian and endometrial malignancy should be taken into consideration during evaluation of patients with endometriosis and breast cancer histories. PMID- 17920405 TI - Ethical issues surrounding the cryopreservation of human oocytes. PMID- 17920407 TI - An expert forum for the histology of endometriomas. PMID- 17920409 TI - Perspectives on oocyte research. PMID- 17920411 TI - Management of long-bone non-unions. PMID- 17920412 TI - Risk factors contributing to fracture non-unions. PMID- 17920413 TI - Definition and classification of fracture non-unions. AB - Classifications in general provide relevant information for clinical purposes to compose a suitable treatment strategy and for research purposes to be able to define comparable study groups. Two distinct types of non-unions are described in the established literature. In the first type the ends of the fragments are hypervascutar or hypertrophic and are capable of biologic reaction. In the second type the ends of the fragments are avascular or atrophic and are inert and incapable of biologic reaction. Hypervascular as well as avascular non-unions may be complicated by the presence of infection, poor soft-tissue quality, short peri articular fragments or significant deformity, demanding mu[ti-stage treatment strategies with concomitant worsened prognosis and subsequent increased frequency of amputation. PMID- 17920414 TI - Principles of management of septic non-union of fracture. PMID- 17920415 TI - Prevalence of long-bone non-unions. AB - Despite the enormous progress made during recent decades in the treatment of long bone fractures, fracture healing is still haunted by complications and above all non-unions. Non-unions represent a particular challenge, and the difficulties surrounding their management are frequently underestimated. Knowledge of the epidemiology of long-bone non-union can assist the treating surgeon in the application of the optimum fracture treatment. PMID- 17920416 TI - The value of laboratory and imaging studies in the evaluation of long-bone non unions. AB - Delayed union and non-union are common but definitive diagnosis is difficult. Few studies have evaluated imaging methods and laboratory parameters in the distinction between normal and prolonged heating specifically of long-bone fractures. The values of newer imaging methods have been analysed in some cases and should be the subject of further clinical trials. Biological markers may facilitate the earliest diagnosis of delayed union. Preliminary studies of these are limited and more trials are needed. To date it is still difficult to predict accurately a normal consolidation of bone. PMID- 17920417 TI - Humeral diaphyseal aseptic non-unions: an Algorithm of management. AB - Successful operative treatment of a humeral shaft non-union may be a challenge for the surgeon. Several treatment options have been reported over the years. A systematic review of the literature was performed. Twenty-three retrieved articles (level of evidence IV) fulfilled our inclusion criteria and dealt with aseptic diaphyseal humeral non-union managed by plating, intramedullary nailing and external fixation based on Ilizarov's principles. Despite an obvious superiority of plating in the treatment of humeral shaft non-unions, there is no doubt that intramedullary nailing as well as external fixation devices have a role. An algorithm of management of the humeral shaft non-unions following a rational approach is suggested. PMID- 17920418 TI - Femoral neck non-unions: how do I do it? AB - Femoral head necrosis and non-union are frequent complications after femoral neck fracture. The main reason for failure leading to non-union is an inadequate osteosynthesis and/or poor mechanical conditions, leading to instability. Criteria for optima reduction and fixation techniques, which can prevent non union in the majority of cases, are described. This knowledge is mandatory for each surgeon as in the non-expert situation up to 30% inadequacy of the "simple" procedure occurs! Although in the elderly endoprosthetic replacement is the treatment of first choice, in the younger and active patients the treatment should be directed towards salvage of the own hip. In non-complex cases a valgisation osteotomy according to Pauwels will lead to very good results. The technique of this secondary procedure is demonstrated by a case report. In case of combined pathology with (complete) a vascular necrosis of the femoral head, the age threshold for endoprosthetic replacement will be far lower nowadays, but even in those cases, especially below the age of 50, salvage procedures with free fibular grafting lead to a good outcome and form a useful alternative. PMID- 17920419 TI - Femoral diaphyseal aseptic non-unions: is there an ideal method of treatment? AB - Femoral non-unions represent a formidable challenge for the orthopaedic surgeon. Their successful treatment is frequently time consuming, and requires utilisation of numerous resources. Three main methods of treatment have been described: intramedullary nailing (IMN), plating and external fixation. A systematic review of the existing English literature of femoral diaphyseal non-union treatment methods has been conducted. The gold standard remains exchange nailing despite the fact that plating has reached near equivocal rates of success. In cases where exchange nailing fails, the use of plates and external fixators has been shown to provide useful adjuncts to the nail. Most surgeons have preserved bone grafting as an option at a secondary or tertiary stage, after the initial revision procedure has failed. PMID- 17920420 TI - Biological enhancement of tibial diaphyseal aseptic non-unions: the efficacy of autologous bone grafting, BMPs and reaming by-products. AB - The mandatory stimulus that can optimise the healing pathway can be electrical, mechanical, biological, or a combination of all these parameters. A variety of means has been utilised for biological enhancement, including extracorporeal shock wave, electrical, ultrasound stimulation, the reaming technique of IM nailing, bone graft substitutes, osteogenic cells and bioactive molecules produced by tissue engineering techniques. The aim of this study is to present a review of the existing evidence for the efficacy of reaming, autologous bone grafting and the commercially available growth factors (BMP-2 and BMP-7) for the treatment of aseptic tibial non-unions. The gold standard method of enhancing bone healing in cases of tibial non-union remains the autologous bone graft. Autogenous bone grafts possess osteoconductive, osteoinductive properties and also osteoprogenitor cells. However, their harvesting is associated with high morbidity and many complications reaching percentages of 30%. Intramedullary reamed nailing, either used as an alternative fixation method or as an exchange to a wider implant, offers the unique biomechanical advantages of an intramedullary device, together with the osteoinductive stimulus of the by products of reaming, and the aptitude for early weight-bearing and active rehabilitation. The safety of administration of the commercial distributed growth factors (BMP-2 and BMP-7), combined with the lack of the morbidity and the quantity restrictions that characterise autologous bone grafts, have given to this family of molecules a principal role between the other bone graft substitutes. On average the union rates reported in the 20 manuscripts that have been evaluated range from 58.3% to 100%, and the average time to union from 12.5 weeks to 48.4 weeks, indicating the significant discrepancies in the reported evidence and the multiplicity of different treatment strategies. PMID- 17920421 TI - The health economics of the treatment of long-bone non-unions. AB - A review of the existing evidence on economic costs of treatment of long-bone fracture non-unions has retrieved 9 papers. Mostly the tibial shaft non-unions have been utilised as models for these economic analyses. Novel treatment strategies like BMP-7 grafting, Ilizarov ring external fixation or supplementary use of therapeutic ultrasound devices have been compared with standard methods of treatment focusing on direct and indirect costs and expenses. A cost identification query was conducted and revealed costs of pound 15,566, pound 17,200 and pound 16,330 for humeral, femoral, and tibial non-unions respectively on a "best-case scenario". The existing scientific evidence can only imply the extent of the economic burden of long-bone non-unions. Further systematic studies are needed to assess the direct medical, direct non-medical, indirect, and monetised quality of life and psychosocial costs of non-unions. PMID- 17920423 TI - Unaltered development of the archi- and neocortex in prematurely born infants: genetic control dominates in proliferation, differentiation and maturation of cortical neurons. AB - The development of cerebral cortex includes highly organized, elaborate and long lasting series of events, which do not come to an end by the time of birth. Indeed, many developmental events continue after the 40th postconceptual week resulting in a long morphological, behavioral and cognitive development of children. Premature birth causes an untimely dramatic change in the environment of the human fetus and often results in serious threats for life. Cognitive abilities of prematurely born children vary, but a correlation between cognitive impairment and the time of birth is evident. In this study we review the morphological evidence of cortical maturation in preterm and full-term infants. Various aspects of postnatal cortical development including cell proliferation and maturation of neurons in the temporal archi- and neocortex are discussed and compared in preterm infants and age-matched full-term controls. Our results suggest that cell proliferation and maturation are not influenced by the preterm delivery. In contrast, the perinatal decrease of the number of Cajal-Retzius cells might be regulated by a mechanism that is affected by preterm birth. We demonstrate that cognitive deficiencies of the prematurely born infants cannot be explained with light microscopically observed alteration of proliferation and maturation of neurons. PMID- 17920424 TI - Subcortical regulation of cortical development: some effects of early, selective deprivations. AB - Selective deprivations, such as the lack of sensory input, of social contacts and of language during the critical (sensitive) period of brain development have profound consequences for the structure and function of the adult brain. The field is largely uncharted since only the consequences of the most severe forms of deprivation are known, and that too only in a few systems. It is similarly unknown if the opposite of deprivation, selective over-stimulation in development, which appears to enhance the acquisition of certain skills, for example musical skills, has collateral deprivation-like effects in other domains. In spite of these uncertainties, I propose that the common mechanism underlying the effects of deprivation may be the altered stabilization of neuronal morphologies, particularly connectivity, in the period when their exuberant development is down regulated. PMID- 17920425 TI - The mirror-neurons system: data and models. AB - In this chapter we discuss the mirror-neurons system, a cortical network of areas that enables individuals to understand the meaning of actions performed by others through the activation of internal representations, which motorically code for the observed actions. We review evidence indicating that this capability does not depend on the amount of visual stimulation relative to the observed action, or on the sensory modality specifically addressed (visual, acoustical). Any sensorial cue that can evoke the "idea" of a meaningful action activates the vocabulary of motor representations stored in the ventral premotor cortex and, in humans, especially in Broca's area. This is true also for phonoarticulatory actions, which determine speech production. We present also a model of the mirror-neurons system and its partial implementation in a set of two experiments. The results, according to our model, show that motor information plays a significant role in the interpretation of actions and that a mirror-like representation can be developed autonomously as a result of the interaction between the individual and the environment. PMID- 17920426 TI - Apraxia: a review. AB - Praxic functions are frequently altered following brain lesion, giving rise to apraxia - a complex pattern of impairments that is difficult to assess or interpret. In this chapter, we review the current taxonomies of apraxia and related cognitive and neuropsychological models. We also address the questions of the neuroanatomical correlates of apraxia, the relation between apraxia and aphasia and the analysis of apraxic errors. We provide a possible explanation for the difficulties encountered in investigating apraxia and also several approaches to overcome them, such as systematic investigation and modeling studies. Finally, we argue for a multidisciplinary approach. For example, apraxia should be studied in consideration with and could contribute to other fields such as normal motor control, neuroimaging and neurophysiology. PMID- 17920428 TI - Visual tracking and its relationship to cortical development. AB - Measurements of visual tracking in infants have been performed from 2 weeks of age. Although directed appropriately, the eye movements are saccadic at this age. Over the first 4 months of life, a rapid transition to successively smoother eye movements takes place. Timing develops first and at 7 weeks of age the smooth pursuit is well timed to a sinusoidal motion of 0.25 Hz. From this age, the gain of the smooth pursuit improves rapidly and from 4 months of age, smooth pursuit dominates visual tracking in combination with head movements. This development reflects massive cortical and cerebellar changes. The coordination between eyes head-body and the external events to be tracked presumes predictive control. One common type of model for explaining the acquisition of such control focuses on the maturation of the cerebellar circuits. A problem with such models, however, is that although Purkinje cells and climbing fibers are present in the newborn, the parallel and mossy fibers, essential for predictive control, grow and mature at 4-7 months postnatally. Therefore, an alternative model that also includes the prefrontal cerebral cortex might better explain the early development of predictive control. The prefrontal cortex functions by 3-4 months of age and provides a site for prediction of eye movements as a part of cerebro-cerebellar nets. PMID- 17920427 TI - Effects of early visual deprivation on perceptual and cognitive development. AB - During early infancy, visual capabilities are quite limited. Nevertheless, patterned visual input during this period is necessary for the later development of normal vision for some, but not all, aspects of visual perception. The evidence comes from studies of children who missed early visual input because it was blocked by dense, central cataracts in both eyes. In this article, we review the effects of bilateral congenital cataracts on two aspects of low-level vision- acuity and contrast sensitivity, and on three aspects of higher-level processing of faces. We end by discussing the implications for understanding the developmental mechanisms underlying normal perceptual and cognitive development. PMID- 17920429 TI - Visual and visuocognitive development in children born very prematurely. AB - Preterm birth is a risk factor for deficits of neurological and cognitive development. Four cohort studies are reported investigating the effects of very premature birth (<32 weeks gestation) on visual, visuocognitive and visuomotor function between birth and 6-7 years of age. The first study used two measures of early visual cortical function, orientation reversal visual event-related potentials (OR-VERP) and fixation shifts under competition. Both these functional measures of visual development correlated with the severity of brain abnormality observed on structural MRI at and before term, and were sensitive predictors of neurodevelopmental outcome at 2 years. The second study compared VERP measures for orientation-reversal and direction-reversal (DR) stimuli, from 2 to 5 months post-term age, in healthy very premature infants compared to infants born at term. The groups did not differ on the development of OR-VERP responses, but the development of the DR-VERP motion responses was delayed in the premature group despite the absence of any brain damage visible on ultrasound, consistent with the developmental vulnerability we have identified in the dorsal cortical stream. The third study used the Atkinson Battery of Child Development for Examining Functional Vision (ABCDEFV) to assess sensory, perceptual, cognitive and spatial visual functions, together with preschool tests of attention and executive function. The premature group showed delays on these tests in line with severity of observed perinatal brain damage on structural MRI at term age. Deficits on certain spatial tasks (e.g. block-construction copying) and executive function tests (e.g. the detour box task) were apparent even in children with minimal damage apparent on MRI. The fourth study tested a large cohort of 6- to 7-year old children born before 32 weeks gestation, across a wide range of cognitive domains, including new tests of spatial cognition and memory. The premature group as a whole showed significant deficits on both auditory and visual tests of attention and attentional control from the TEA-Ch battery, on tests of location memory, block construction and on many visuocognitive and visuomotor tests. Development was generally relatively normal on language tests and on WPPSI scores. Factor analysis showed that while general cognitive ability accounted for the largest part of the variance, significant deficits, and a relationship to MRI results, were primarily in spatial, motor, attention and executive function tests. A model is proposed suggesting that the cluster of deficits seen in children born prematurely may be related to networks involving the cortical dorsal stream and its connections to parietal, frontal and hippocampal areas. PMID- 17920430 TI - Development of brain mechanisms for visual global processing and object segmentation. AB - Objects have specific cognitive attributes, elicit particular visuo-motor responses, and require visual processes beyond primary visual cortex to combine information over extended regions as a basis for the segmentation and integration of visual objects. As well as segmentation, the assignment of region boundaries to differentiate figure from ground is a key process whose operation can be observed in infants during the early months. Global organization of both motion and pattern information plays a role in object segmentation and integration. These two types of global processing are associated with different brain systems, have different developmental courses, and are differentially vulnerable in developmental disorders. In infancy, specific visual attributes determine the selection of a manual response (reach-and-grasp vs. surface exploration) and also the detailed kinematic parameters of each class of response. These are taken to reflect the properties of distinct visuo-motor modules whose properties emerge between 4 and 12 months of age. While these modules are a component of the dorsal cortical stream, they must interact with ventral stream processing in development and in the mature system. PMID- 17920432 TI - The early development of visual attention and its implications for social and cognitive development. AB - Looking behavior plays a crucial role in the daily life of an infant and forms the basis for cognitive and social development. The infant's visual attentional systems undergo rapid development during the first few months of life. During the last decennia, the study of visual attentional development in infants has received increasing interest. Several reliable measures to investigate the early development of attentional processes have been developed, and currently a number of new methods are giving fresh impetus to the field. Research on overt and covert as well as exogenously and endogenously controlled attention shifts is presented. The development of gaze shifts to peripheral targets, covert attention, and visual scanning behavior is treated. Whereas most attentional mechanisms in very young infants are thought to be mediated mainly by subcortical structures, cortical mechanisms become increasingly more functional throughout the first months. Different accounts of the neurophysiological underpinnings of attentional processes and their developmental changes are discussed. Finally, a number of studies investigating the implications of attentional development for early cognitive and social development are presented. PMID- 17920431 TI - How face specialization emerges in the first months of life. AB - The present chapter deals with the topic of the ontogeny and development of face processing in the first months of life and is organized into two sections concerning face detection and face recognition. The first section focuses on the mechanisms underlying infants' visual preference for faces. Evidence is reviewed supporting the contention that newborns' face preferences is due to a set of non specific constraints that stem from the general characteristics of the human visuo-perceptual system, rather than to a representational bias for faces. It is shown that infants' response to faces becomes more and more tuned to the face category over the first 3 months of life, revealing a gradual progressive specialization of the face-processing system. The second section sought to determine the properties of face recognition at birth. In particular, a series of experiments are presented to examine whether the inner facial part is processed and encoded when newborns recognize a face, and what kind of information- featural or configural--newborns' face recognition rely on. Overall, results are consistent with the existence of general constraints present at birth that tune the system to become specialized for faces later during development. PMID- 17920433 TI - Visual constraints in the development of action. AB - The chapter's aim is to understand the role of visual information in the control of avoidance and interception behaviors in infancy from the ecological psychology approach to perception and action. We show that during infancy developmental change in action is associated with the use of different information sources and that this process of attunement promotes the perceived action possibilities (affordances). In the final section, we position these findings within Milner and Goodale's two-visual system model, which holds that perception and action are mediated by two functionally and neuron-anatomically separate visual (sub )systems. PMID- 17920434 TI - Object and event representation in toddlers. AB - Mental representation of absent objects and events is a major cognitive achievement. Research is presented that explores how toddlers (2- to 3-year-old children) search for hidden objects and understand out-of-sight events. Younger children fail to use visually obvious cues, such as a barrier that blocks a moving object's path. Spatiotemporal information provided by movement cues directly connected to the hidden object is more helpful. A key problem for toddlers appears to be difficulty in representing a spatial array involving events with multiple elements. PMID- 17920435 TI - Learning and development in infant locomotion. AB - The traditional study of infant locomotion focuses on what movements look like at various points in development, and how infants acquire sufficient strength and balance to move. We describe a new view of locomotor development that focuses on infants' ability to adapt their locomotor decisions to variations in the environment and changes in their bodily propensities. In the first section of the chapter, we argue that perception of affordances lies at the heart of adaptive locomotion. Perceiving affordances for balance and locomotion allows infants to select and modify their ongoing movements appropriately. In the second section, we describe alternative solutions that infants devise for coping with challenging locomotor situations, and various ways that new strategies enter their repertoire of behaviors. In the third section, we document the reciprocal developmental relationship between adaptive locomotion and cognition. Limits and advances in means-ends problem solving and cognitive capacity affect infants' ability to navigate a cluttered environment, while locomotor development offers infants new opportunities for learning. PMID- 17920436 TI - Core systems in human cognition. AB - Research on human infants, adult nonhuman primates, and children and adults in diverse cultures provides converging evidence for four systems at the foundations of human knowledge. These systems are domain specific and serve to represent both entities in the perceptible world (inanimate manipulable objects and animate agents) and entities that are more abstract (numbers and geometrical forms). Human cognition may be based, as well, on a fifth system for representing social partners and for categorizing the social world into groups. Research on infants and children may contribute both to understanding of these systems and to attempts to overcome misconceptions that they may foster. PMID- 17920437 TI - Taking an action perspective on infant's object representations. AB - At around 4 months of age, infants predict the reappearance of temporary occluded objects. Younger infants have not demonstrated such an ability, but they still benefit from experience; decreasing their reactive saccade latencies over successive passages from the earliest age tested (7 weeks of age). We argue that prediction is not an all or none process that infants either lack or possess. Instead, the ability to predict the reappearance of an occluded object is dependent on numerous simultaneous factors, including the occlusion duration, the manner in which the object disappears, and previous experiences with similar events. Furthermore, we claim that infants' understanding of how occluded objects move is based on prior experiences with similar events. Initially, infants extrapolate occluded object motion, because they have massive experience with such motion. But infants also have the ability to rapidly adjust to novel trajectories that violate their initial expectations. All of these findings support a constructivist view of infants object representations. PMID- 17920438 TI - Infants' perception and production of intentional actions. AB - Human beings act and interact with their social environment. Thus, it is important not only to understand other individuals' actions, but also to control one's own actions. To understand intentional actions one needs to detect goals in the perceived actions of others as well as to control one's own movements in order to achieve these goals through action production. After a short review of recent studies on the development of action understanding during the first years of life, the role of action effects for action understanding is discussed. In a series of experiments the exchange between action perception and action production is demonstrated, its implications for understanding intentional actions are highlighted. PMID- 17920439 TI - The role of behavioral cues in understanding goal-directed actions in infancy. AB - Infants show very early sensitivity to a variety of behavioral cues (such as self propulsion, equifinal movement, free variability, and situational adjustment of behavior) that can be exploited when identifying, predicting, and interpreting goal-directed actions of intentional agents. We compare and contrast recent alternative models concerning the role that different types of behavioral cues play in human infants' early understanding of animacy, agency, and intentional action. We present new experimental evidence from violation of expectation studies to evaluate these alternative models on the nature of early development of understanding goal-directedness by human infants. Our results support the view that, while infants initially do not restrict goal attribution to behaviors of agents exhibiting self-propelled motion, they quickly develop such expectations. PMID- 17920440 TI - Seeing the face through the eyes: a developmental perspective on face expertise. AB - Most people are experts in face recognition. We propose that the special status of this particular body part in telling individuals apart is the result of a developmental process that heavily biases human infants and children to attend towards the eyes of others. We review the evidence supporting this proposal, including neuroimaging results and studies in developmental disorders, like autism. We propose that the most likely explanation of infants' bias towards eyes is the fact that eye gaze serves important communicative functions in humans. PMID- 17920441 TI - Past and present challenges in theory of mind research in nonhuman primates. AB - This paper presents the trajectory of theory of mind research in nonhuman primates, with a special focus on chimpanzees as they have been the most intensely studied species. It analyzes the main developments in the field, the critiques that they raised, the responses that they have generated and the current challenges faced by the field. Currently, the most plausible working hypothesis is that at least chimpanzees know what others can and cannot see. Using tasks with a high ecological validity and mapping out key concepts such association and inference are postulated as fundamental steps to further advance our knowledge in this area. PMID- 17920442 TI - Infancy and autism: progress, prospects, and challenges. AB - We integrate converging evidence from a variety of research areas in typical and atypical development to motivate a developmental framework for understanding the emergence of autism in infancy and to propose future directions for a recent area of research focusing on infant siblings of children with autism. Explaining the cognitive profile in autism is best achieved through tracing the process through which associated symptoms emerge over development. Understanding this process would shed light on the underlying causes of this multifaceted condition through clarifying how and why a variety of risk factors, single or in combination, exert an impact on the resulting phenotype. We emphasize the importance of integrating theoretical models of typical development in understanding atypical development and argue for the need to develop continuous and individually valid measures for at-risk infants both for predictors and outcomes of autism symptoms. PMID- 17920443 TI - Children-robot interaction: a pilot study in autism therapy. AB - We present here a pilot study of child-robot interactions, in which we discuss developmental origins of human interpersonal communication. For the past few years, we have been observing 2- to 4-year-old children with autism interacting with Keepon, a creature-like robot that is only capable of expressing its attention (directing its gaze) and emotions (pleasure and excitement). While controlled by a remote experimenter, Keepon interacted with the children with its simple appearance and actions. With a sense of curiosity and security, the children spontaneously approached Keepon and engaged in dyadic interaction with it, which then extended to triadic interactions where they exchanged with adult caregivers pleasure and surprise they found in Keepon. Qualitative and quantitative analysis of these unfolding interactions suggests that autistic children possess the motivation to share mental states with others, which is contrary to the commonly held position that this motivation is impaired in autism. We assume Keepon's minimal expressiveness helped the children understand socially meaningful information, which then activated their intact motivation to share interests and feelings with others. We conclude that simple robots like Keepon would facilitate social interaction and its development in autistic children. PMID- 17920444 TI - Sensorimotor coordination in a "baby" robot: learning about objects through grasping. AB - This paper describes a developmental approach to the design of a humanoid robot. The robot, equipped with initial perceptual and motor competencies, explores the "shape" of its own body before devoting its attention to the external environment. The initial form of sensorimotor coordination consists of a set of explorative motor behaviors coupled to visual routines providing a bottom-up sensory-driven attention system. Subsequently, development leads the robot from the construction of a "body schema" to the exploration of the world of objects. The "body schema" allows controlling the arm and hand to reach and touch objects within the robot's workspace. Eventually, the interaction between the environment and the robot's body is exploited to acquire a visual model of the objects the robot encounters which can then be used to guide a top-down attention system. PMID- 17920445 TI - Emergence and development of embodied cognition: a constructivist approach using robots. AB - A constructivist approach to cognition assumes the minimal and the simplest set of initial principles or mechanisms, embeds them in realistic circumstances, and lets the entire system evolve under close observation. This paper presents a line of research along this approach trying to connect embodiment to social cognition. First, we show that a mere physical body, when driven toward some task goal, provides a clear information structure, for action execution and perception. As a mechanism of autonomous exploration of such structure, "embodiment as a coupled chaotic field" is proposed, with experiments showing emergent and adaptive behavior. Scaling up the principles, a simulation of the fetal/neonatal motor development is presented. The musculo-skeletal system, basic nervous system, and the uterus environment are simulated. The neural-body dynamics exhibit spontaneous exploration of a variety of motor patterns. Lastly, a robotic experiment is presented to show that visual-motor self-learning can lead to neonatal imitation. PMID- 17920447 TI - Self-awareness and the left inferior frontal gyrus: inner speech use during self related processing. AB - To test the hypothesis of a participation of inner speech in self-referential activity we reviewed 59 studies measuring brain activity during processing of self-information in the following self-domains: agency, self-recognition, emotions, personality traits, autobiographical memory, preference judgments, and REST. The left inferior frontal gyrus (LIFG) has been shown to sustain inner speech use. We calculated the percentage of studies reporting LIFG activity for each self-dimension. 55.9% of all studies reviewed identified LIFG (and presumably inner speech) activity during self-awareness tasks. Furthermore, the LIFG was more frequently recruited during conceptual tasks (e.g., emotions, traits) than during perceptual tasks (e.g., agency, self-recognition). This supports the view of a relative involvement of inner speech in self-reflective processes. PMID- 17920446 TI - Thymic selection stifles TCR reactivity with the main chain structure of MHC and forces interactions with the peptide side chains. PMID- 17920448 TI - Vesicular glutamate transporter 2 protein and mRNA containing neurons in the hypothalamic suprachiasmatic nucleus of the rat. AB - The hypothalamic suprachiasmatic nucleus is the key structure of the control of circadian rhythms and has a rich glutamatergic innervation. Besides the presence of glutamatergic afferents, several findings also suggest the existence of glutamatergic efferents from the suprachiasmatic nucleus to its target neurons in various prominent hypothalamic cell groups. However, there is no direct neuromorphological evidence for the presence of glutamatergic neurons in the suprachiasmatic nucleus. Therefore, the purpose of the present investigations was to try to clarify this question. Immunocytochemistry was used at the light and electron microscopy level to identify vesicular glutamate transporter type 2 (VGluT2) immunopositive (presumed glutamatergic) neurons in the rat suprachiasmatic nucleus. In addition VGluT2 mRNA expression in neurons of the nucleus was also addressed with radioisotopic in situ hybridization. Both at the light and electron microscopy level we detected VGluT2 positive neurons, which did not contain GABA, vasoactive intestinal polypeptide or vasopressin. Further, we demonstrated the expression of VGluT2 mRNA in a few cells within the suprachiasmatic nucleus; these glutamatergic cells were distinct from somatostatin mRNA expressing neurons. As VGluT2 is a selective marker of glutamatergic neuronal elements, the present observations provide direct neuromorphological evidence for the presence of glutamatergic neurons in the cell group. PMID- 17920449 TI - Manganese intoxication decreases the expression of manganoproteins in the rat basal ganglia: an immunohistochemical study. AB - Manganese (Mn) is a cofactor for some metalloprotein enzymes, including Mn superoxide dismutase (Mn-SOD), a mitochondrial enzyme predominantly localized in neurons, and glutamine synthetase (GS), which is selectively expressed in astroglial cells. The detoxifying effects of GS and Mn-SOD in the brain, involve catabolizing glutamate and scavenging superoxide anions, respectively. Mn intoxication is characterized by impaired function of the basal ganglia. However, it is unclear whether regional central nervous system expression of manganoproteins is also affected. Here, we use immunocytochemistry in the adult rat brain, to examine whether Mn overload selectively affects the expression of GS, Mn-SOD, Cu/Zn-SOD, another component of the SOD family, and glial fibrillary acid protein (GFAP), a specific marker of astrocytes. After chronic Mn overload in drinking water for 13 weeks, we found that the number and immunostaining intensity of GS- and Mn-SOD-positive cells was significantly decreased in the striatum and globus pallidus, but not in the cerebral frontal cortex. In addition, we found that GS enzymatic activity was decreased in the strio-pallidal regions but not in the cerebral cortex of Mn-treated animals. In contrast, Cu/Zn SOD- and GFAP-immunoreactivity was unchanged in both the cerebral cortex and basal ganglia of Mn-treated rats. Thus, we conclude that in response to chronic Mn overload, a down-regulation of some manganoproteins occurs in neurons and astrocytes of the striatum and globus pallidus, probably reflecting the vulnerability of these regions to Mn toxicity. PMID- 17920450 TI - Evidence for hemispheric specialization in the marmoset (Callithrix penicillata) based on lateralization of behavioral/neurochemical correlations. AB - A correlative study between behavioral, neurochemical and hormonal measures was conducted on male black tufted-ear marmoset monkeys (Callithrix penicillata). Behavioral analysis was performed in order to examine the effects of confrontation with a natural predator (taxidermized oncilla cat, Felis tigrina). The subjects were subjected to four trials without predator, six confrontation trials with predator present, and four trials with the predator removed. Handedness was analyzed by the frequency with which they performed scratching, grooming and hanging behaviors with the left or right hands. The animals' brains were subjected to ex vivo neurochemical analysis of several structures from both hemispheres. The content of monoamines, acetycholine and metabolites were analyzed by HPLC-ED. Plasma levels of cortisol and adrenocorticotrophic hormone (ACTH) were analyzed by chemoluminescence immunoassay. Testosterone plasma concentration was determined by radioimmunoassay. Higher levels of dopamine and acetylcholine were detected in the right caudate/putamen, in comparison to the left. For the remaining areas, similar levels were observed in both hemispheres. A hand preference between and within the behaviors scored was not detected. However, correlative analyses revealed complex interactions between the behavioral and neurochemical measures, particularly in the left hemisphere. Lateralized correlations were found in relation to brain site, type of behavior, neurochemical parameter and treatment condition, thus providing evidence for functional brain asymmetries in this species. Interhemispheric comparisons of neurochemical/behavioral correlations appear to be a promising approach towards delineating hemispheric specialization of functions in this, and perhaps, other species. PMID- 17920451 TI - Characterization of an immunodeficient mouse model of mucopolysaccharidosis type I suitable for preclinical testing of human stem cell and gene therapy. AB - Mucopolysaccharidosis type I (MPS-I or Hurler syndrome) is an inherited deficiency of the lysosomal glycosaminoglycan (GAG)-degrading enzyme alpha-l iduronidase (IDUA) in which GAG accumulation causes progressive multi-system dysfunction and death. Early allogeneic hematopoietic stem cell transplantation (HSCT) ameliorates clinical features and extends life but is not available to all patients, and inadequately corrects its most devastating features including mental retardation and skeletal deformities. To test novel therapies, we characterized an immunodeficient MPS-I mouse model less likely to develop immune reactions to transplanted human or gene-corrected cells or secreted IDUA. In the liver, spleen, heart, lung, kidney and brain of NOD/SCID/MPS-I mice IDUA was undetectable, and reduced to half in heterozygotes. MPS-I mice developed marked GAG accumulation (3-38-fold) in these organs. Neuropathological examination showed GM(3) ganglioside accumulation in the striatum, cerebral peduncles, cerebellum and ventral brainstem of MPS-I mice. Urinary GAG excretion (6.5-fold higher in MPS-I mice) provided a non-invasive and reliable method suitable for serially following the biochemical efficacy of therapeutic interventions. We identified and validated using rigorous biostatistical methods, a highly reproducible method for evaluating sensorimotor function and motor skills development. This Rotarod test revealed marked abnormalities in sensorimotor integration involving the cerebellum, striatum, proprioceptive pathways, motor cortex, and in acquisition of motor coordination. NOD/SCID/MPS-I mice exhibit many of the clinical, skeletal, pathological and behavioral abnormalities of human MPS-I, and provide an extremely suitable animal model for assessing the systemic and neurological effects of human stem cell transplantation and gene therapeutic approaches, using the above techniques to measure efficacy. PMID- 17920452 TI - Co-expression of myosin II regulatory light chain and the NMDAR1 subunit in neonatal and adult mouse brain. AB - Movement of glutamate receptors in neurons likely involves direct and indirect association of receptor subunits with microtubule- and actin-based motor proteins. We have previously shown that myosin II regulatory light chain (RLC) binds directly to subunits of the NMDA-type glutamate receptor (NR), suggesting that NMDA receptors are closely associated with a myosin II motor complex. Using a polyclonal antibody predicted to recognize all RLC isoforms previously described in rodent brain, we report the expression of RLC and the NR1 subunit in cortex, hippocampus and cerebellum of postnatal day 0 (P0) and adult mouse. Although myosin RLC was not exclusively localized with NR1 by immunohistochemistry, co-staining was striking in the neuronal soma of deep cortical neurons and Purkinje neurons of the cerebellum which showed a punctate, perinuclear pattern of immunoreactivity. These neuronal populations were identified using a monoclonal antibody directed against a nuclear-specific, transcriptional repressor, chicken ovalbumin upstream promoter-transcription factor (COUP-TF)-interacting protein 2 (CTIP2). Co-expression of NR1 and a myosin II motor was validated using an isoform specific anti-nonmuscle myosin II-B heavy chain (NMHC II-B) antibody. Our findings support the idea that there is regional heterogeneity in the molecular composition of the NMDA receptor-associated cytoskeleton, and suggest that NR subunits may be associated with an actin-based, myosin II-B motor within the endomembrane system of some neuronal populations. Differential staining patterns observed with light and heavy chain antibodies, however, suggest that there is also heterogeneity in the composition of myosin II complexes in brain. PMID- 17920454 TI - Evaluation of the ruminant complete blood cell count. AB - The complete blood cell count can provide valuable diagnostic and prognostic information when coupled with a thorough physical examination. This article addresses proper sample handling, storage, and interpretation of the erythron, serum iron indices, leukon, and acute-phase proteins for cattle, sheep, and goats. PMID- 17920455 TI - Evaluation of the ruminant serum chemistry profile. AB - In general, laboratory tests provide useful information about a patient and aid in making a diagnosis, determining a prognosis, formulating a treatment plan, and monitoring response. The serum chemistry profile, composed of a battery of tests, is readily available to clinicians through in-house testing or veterinary reference and diagnostic laboratories. A chemistry profile allows for evaluation of several body systems and assessment of metabolic, acid-base, or electrolyte disturbances. The first sections of this article introduce concepts important to all laboratory tests, such as sample collection and handling, reference intervals, and quality control. The remainder of the article discusses each of the analytes found on a typical clinical chemistry profile and the interpretation of abnormal results. PMID- 17920456 TI - Field chemistry analysis. AB - Typical manual and automated technologies used in field chemistry testing are reviewed, along with associated advantages and disadvantages. A brief overview of metabolic disease monitoring is included. Guidelines for evaluating and achieving success are provided, including criteria for system evaluation and expectations for comparative performance evaluations. The more common problems and limitations associated with field chemistry diagnostics and how to best prevent them are also discussed. PMID- 17920457 TI - Cytology in food animal practice. AB - Diagnostic cytology can greatly aid the clinician in determining a more refined diagnosis, prognosis, and treatment plan to serve the client and patient better. Sample collection is not difficult and can be done in the field as well as in a hospital setting. The collection and sample handling procedures described in this article can help the clinician to obtain diagnostically valuable samples. In many cases, preliminary cytologic evaluation can be performed by the general practitioner. Additional diagnostic evaluation and interpretation are readily available from trained pathologists at diagnostic laboratories. PMID- 17920458 TI - Molecular diagnostics for the food animal practitioner. AB - Molecular diagnostics are becoming widely used as routine diagnostic tests performed by food animal practitioners. This article discusses the variations of several commonly performed molecular assays with regard to their molecular basis and the appropriate interpretation of the results. Applications of these methods are discussed in the context of infectious disease testing and genetic testing of food animal species. PMID- 17920459 TI - Performing the field necropsy examination. AB - This article is designed to aid the practitioner by maximizing the effectiveness of field postmortem diagnostic investigations. Contents include an outline of the procedure for field necropsy of ruminants, recommended tools and supplies, and guidelines for sample collection and submission. PMID- 17920460 TI - Diagnostic considerations for evaluating nutritional problems in cattle. AB - The advances in testing methodology and in our understanding of the normal nutritional physiology of cattle have given veterinarians valuable tools to assess the nutritional status of cattle. By taking a representative number of appropriate samples, it is now relatively easy and inexpensive to determine the nutritional status of an individual or a group of animals. Also, the effectiveness of supplementation programs can be measured over time as part of an overall preventive medicine program. PMID- 17920461 TI - Diagnostic ultrasonography in ruminants. AB - The use of diagnostic ultrasound equipment is becoming widespread within various sectors of veterinary practice. Ultrasonographic examination has several advantages over other imaging modalities and it can be applied in hospital and ambulatory settings. It has the potential for widespread use in the diagnosis of disorders of several body systems in food animal species. This article describes its application to gastrointestinal, hepatobiliary, cardiothoracic, urogenital, and umbilical disorders. Normal and abnormal findings are portrayed and discussed. Accurate and timely use of this diagnostic modality requires a modest amount of training and practice, but it allows for relatively rapid, inexpensive, and noninvasive acquisition of clinically relevant data. PMID- 17920462 TI - Pulmonary arterial pressure testing for high mountain disease in cattle. AB - High mountain or brisket disease is an economically costly disease of cattle raised at elevations greater than 1500 m (5000 ft). It appears that no one breed is resistant to the effects of high-altitude hypoxia. Some breeds, and pedigrees within breeds, appear to be more naturally resistant to the effects of high altitude. Multiple factors contribute to the variance in pulmonary arterial pressure in cattle, including breed, gender, body condition, concurrent illness, environmental conditions, elevation, and genetics. Pulmonary arterial pressure testing is an effective diagnostic and management tool used to identify clinically affected and high-risk animals. The procedure can be performed in the field and is an economically valuable method for the selection and breeding management of beef cattle raised at high altitude. PMID- 17920463 TI - Arthroplasty in Asia-Pacific Region. Foreword. PMID- 17920464 TI - Anatomic hip range of motion after implantation during total hip arthroplasty as measured by a navigation system. AB - Simulation of prosthetic impingement is important for preventing complications after total hip arthroplasty (THA). Although the anatomical hip range of motion (ROM) in patients after THA is an essential parameter for these simulations, previous simulation studies substituted various clinical hip ROMs for the anatomical hip ROM. Using a navigation system, anatomical hip ROM was accurately assessed after implantation during primary THA in 30 patients. We found that the hip could be passively moved to 113 degrees of flexion, 34 degrees of extension, 46 degrees of abduction, 75 degrees of internal rotation, and 36 degrees of external rotation. Almost all reference hip ROMs used in previous simulations were smaller than these values. Therefore, wider hip ROM values should be used as parameters for such simulations. PMID- 17920465 TI - Comparison of limb and component alignment using computer-assisted navigation versus image intensifier-guided conventional total knee arthroplasty: a prospective, randomized, single-surgeon study of 467 knees. AB - Total knee arthroplasty was performed in 282 knees with image-free navigation (group A) and in 185 with optimized conventional technique (group B). Mean postoperative mechanical axis of the limb was 179.7 degrees in group A and 179.1 degrees in group B (P < .002). There was a higher percentage of knees in group A that had restoration of mechanical axis to +/-1 degrees, +/-2 degrees, and +/-3 degrees of neutral (P < .0001). There were 9.2% outliers (+/-3 degrees) in group A and 21.6% outliers in group B (P < .0001). For knees exceeding 20 degrees varus, there was no significant difference between the mean mechanical axes in the 2 groups. Both components were aligned within 3 degrees of neutral in 90.8% of the knees in group A and 76.2% of the knees in group B (P < .0001). PMID- 17920466 TI - Gene expression profile of macrophage-like U937 cells in response to polyethylene particles: a novel cell-particle culture system. AB - We investigated the gene expression profiles of U937 cells after contact with polyethylene particles. U937 cells were differentiated with phorbol 12-myristate 13-acetate, and cocultured with either retrieved polyethylene particles or commercially produced polyethylene particles (Ceridust 3615, Clariant Japan, Tokyo, Japan). To achieve consistent contact with the polyethylene particles, we used a rotating device. Phagocytosis of the polyethylene particles or retrieved polyethylene particles by differentiated U937 cells stimulated the release of cytokines including interleukin 1beta, interleukin 6, interleukin 8, and vascular endothelial growth factor. Microarray analysis revealed that the expression of IL8, CCL4, CXCR4, and some other genes was up-regulated after contact with retrieved polyethylene particles. This study first reports the gene expression profiles of U937 cells after contact with polyethylene particles. We believe that this experimental model is applicable to all other particulate materials. PMID- 17920467 TI - Patient sensitivity to polyethylene particles with cemented total hip arthroplasty. AB - To determine whether sensitivity to polyethylene particles varies among patients, we studied 25 patients who had undergone total hip arthroplasty. We used pelvic radiographs to measure annual polyethylene wear and the area of osteolysis. The ratio of the area of osteolysis to the volumetric polyethylene wear was defined as sensitivity index. Adherent cells from peripheral blood were cocultured with polyethylene particles, and the amount of bone-resorptive cytokines was measured. The amount of interleukin-6, but not of interleukin-1beta or tumor necrosis factor-alpha, released from adherent cells in the in vitro experiment correlated with the in vivo sensitivity indices. This technique appears capable of predicting the development of polyethylene-induced osteolysis, allowing surgeons to avoid using polyethylene as the bearing surface in patients at risk for osteolysis. PMID- 17920468 TI - Factors leading to decreased rates of deep vein thrombosis and pulmonary embolism after total knee arthroplasty. AB - We evaluated a total of 473 knees (264 patients) to determine the prevalence of deep vein thrombosis and pulmonary embolism (PE) in those who were not given thromboprophylaxis after total knee arthroplasty (TKA). Deep vein thrombosis was diagnosed by roentgenographic venography, and PE was diagnosed by perfusion lung scanning. In patients with bilateral TKA, 47 (11%) of 418 venograms showed positive findings for thrombi, whereas in patients with unilateral TKA, 11 (20%) of 55 venograms showed positive findings for thrombi (P = .758). No patient had symptoms of PE, and findings for the perfusion lung scans were negative in all patients. We neither treat our patients for deep vein thrombosis and PE prophylactically nor therapeutically unless patient has a symptomatic PE. PMID- 17920469 TI - Computer-aided custom-made hemipelvic prosthesis used in extensive pelvic lesions. AB - We review here our experience in the computer-aided design and manufacture and implantation of custom-made hemipelvic prostheses in 10 patients who underwent internal hemipelvectomy for extensive pelvic lesions. Computed tomography data and the rapid prototyping technique were used to make a precise model of each patient's pelvis for the simulated hemipelvectomy; the model was used to design and manufacture prostheses that were easy to implant because the location and orientation of the acetabulum was readily adjustable. Four patients died 6 to 10 months after surgery; the remaining 6 patients, monitored for 21 to 48 months, all had good hip function. There were early hip dislocations in 2 patients and wound-healing problems in 3, but all were treated successfully. PMID- 17920470 TI - Revision total hip arthroplasty for pelvic osteolysis with well-fixed cementless cup. AB - Treatment of pelvic osteolysis after total hip arthroplasty (THA) remains controversial. Clinical and radiographic outcomes of revision THA were evaluated in 62 hips with pelvic osteolysis and well-fixed cementless cups. The patients' mean age was 50.9 years, and the mean interval from primary to revision THA was 9.7 years. For revision, cementless cups were used in 51 hips, and cemented cups in 11 with acetabular reinforcement rings in 9. The mean duration of follow-up after revision THA was 5.9 years (range, 3.0-9.7 years). At final follow-up, the average Harris Hip Score was 92.4, and there was no radiographic complication except for 1 with change of inclination. Revision THA for pelvic osteolysis with well-fixed cementless cups showed favorable outcomes, and it can be preferentially used in young patients. PMID- 17920471 TI - Revision total hip arthroplasty using a fluted and tapered modular distal fixation stem with and without extended trochanteric osteotomy. AB - We reviewed 62 revision total hip arthroplasties performed using a fluted and tapered modular distal fixation stem after a mean follow-up of 4.2 years. An extended trochanteric osteotomy (ETO) was used in 32 of the 62 hips (52%), whereas no osteotomy was used in the remaining 30 hips. The mean postoperative Harris hip score among the patients was 87.3 points. The mean stem subsidence was 1.1 mm. With the exception of one reoperation for a deep infection, no femoral revision was performed because of mechanical failure. Complications included intraoperative diaphyseal split fractures (6%), cortical perforations (6%), and dislocations (5%). Postoperative Harris hip scores, femoral component stability, and overall complication rates did not differ between the group treated with an ETO and that treated without it. However, the rates of cortical perforation and marked stem subsidence (>5 mm) were significantly higher in the group treated without an ETO than those in the group treated with an ETO, but these were not significantly different when stratified by femoral bone defect. The potential advantages of this implant design could be highlighted in a clinical setting when inserted using an ETO. PMID- 17920472 TI - Cement-within-cement stem exchange using the collarless polished double-taper stem. AB - The clinical and radiographic outcomes of the cement-within-cement femoral stem exchange technique at revision hip arthroplasty were determined. Twenty-three revisions with a collarless polished double-taper stem design were prospectively monitored at up to 12 years. The most common indications for revision were recurrent dislocation and acetabular revision. Radiographic stem subsidence was measured by the Ein Bild Roentgen Analyse method. There was no stem re-revision and no radiographic loosening. Stem within cement subsidence, an intentional design feature of this stem, averaged 0.8 mm (range, 0-2 mm). The average subsidence was similar to that in primary hip arthroplasty. The excellent long term results of cemented collarless polished taper stems for primary and revision hip arthroplasty can be extended even further by cement-within-cement exchange, which preserves the femur. PMID- 17920473 TI - The early clinical outcome of minimally invasive quadriceps-sparing total knee arthroplasty: report of a 2-year follow-up. AB - The results of 32 minimally invasive quadriceps-sparing (Q-S) total knee arthroplasties were compared with those of a matched group of 35 standard total knee arthroplasties. The patients were prospectively followed for a mean of 24 months (range, 18-28 months). The Q-S group was significantly quicker in regaining quadriceps strength and knee flexion and had less pain during the first 2 postoperative weeks. The Knee Society scores showed no significant difference at 6 weeks, 1 year, and last visit. There were 9 outliers in the Q-S group, none in the standard group. The tourniquet time was significantly longer in the Q-S group. Patients in the Q-S group were 100% satisfied about the incision. The Q-S technique showed better and faster recovery, but there were more outliers and bone injuries during surgery, and this coupled with length of tourniquet time were the major disadvantages in our early experience. PMID- 17920474 TI - Progressive quadriceps incision during minimally invasive surgery for total knee arthroplasty: the effect on early postoperative ambulation. AB - A prospective series of 114 consecutive minimally invasive surgeries for total knee arthroplasty was performed using the quadriceps-sparing approach at the beginning. Intraoperatively, when the knee was in 45 degrees to 60 degrees of flexion, lateral patella subluxation was evaluated. A progressive quadriceps tendon incision with a 1-cm increment was applied if the patella could not be completely slid. The mean follow-up time was 24 months. There were 3 groups according to the length of quadriceps incision: group A (17 knees) had no or 1-cm quadriceps incision; group B (60 knees) had 2-cm incision; and group C (37 knees) had 3-cm incision. The average operative time, blood loss, pain score, preoperative range of motion and postoperative range of motion at 2 weeks, 6 weeks, 12 weeks, and 3 months were not significantly different among groups. Patient ability for early ambulation (sitting, knee straightening, standing, and walking) was indifferent between groups A and B; however, this was significantly delayed in group C. In conclusion, minimally invasive surgery for total knee arthroplasty with 2-cm quadriceps incision or strict quadriceps-sparing approach provided no difference on early ambulation. PMID- 17920475 TI - Comparison of hydroxyapatite-coated and non-hydroxyapatite-coated noncemented total hip arthroplasty in same patients. AB - The comparison between standard blasted and hydroxyapatite (HA)-coated cementless femoral stems (CLS stem) was made in the same patients who have bilateral late stage osteonecrosis. Clinical and radiographic evaluations were performed in 20 patients (among 29 patients) with the minimum follow-up of 10 years (range, 123 168 months). Endosteal bone formation was found in all patients of both stems, and no radiolucent line was observed. The mean of initial stem subsidence was 1.27 +/- 0.95 mm with the standard stem and 0.59 +/- 0.56 mm with the HA-coated stem. Other results, such as hip score, endosteal bone formation, cortical hypertrophy, osteolysis, and heterotopic ossification showed no statistical differences. These have suggested that the fixation obtained by the cementless stem with a corundum-blasted surface is stable enough to get excellent clinical and radiographic results without the HA coating. PMID- 17920476 TI - In vitro analysis of exeter stem torsional stability. AB - The effect of cyclic loading on the torsional stiffness of a polished double tapered femoral stem was investigated in vitro. Initial torsional stability was compared with torsional stability after cyclic loading. Stems were removed from the cement mantle and reinserted without the use of additional cement. Torsional stability was measured after reinsertion and after further cyclic loading. Subsidence of the stem was observed. No difference in torsional stiffness was observed during loading. No difference between the stiffness before extraction and after reinsertion was observed. Torsional stiffness of an Exeter stem does not decrease after axial subsidence under cyclic loading. Stability is retained after reinsertion into the original cement mantle. Debonding of the Exeter stem is not associated with rotational instability of the implant. PMID- 17920477 TI - Transverse subtrochanteric shortening osteotomy in primary total hip arthroplasty for patients with severe hip developmental dysplasia. AB - Twenty-four total hip arthroplasties were performed on patients with Crowe grade 3 or 4 hip dysplasia using subtrochanteric shortening osteotomy with 2 kinds of femoral stems. The average age of the patients was 44.8 years, and their average length of follow-up was 4.7 years. Acetabular reconstruction with structural autograft was used in 11 hips. Radiologically, hip centers were nearly normalized by a vertical height of 10.6-mm elevation and a horizontal length of 1.7 mm as compared with uninvolved sites. Three osteotomy nonunions required revisions with bone graft. One acetabular revision was performed for migration. One postoperative dislocation was managed successfully with closed reduction and an abduction brace. However, no neurologic complication was noticed. The Harris hip score improved from 35.6 to 81.7. A cementless modular distal fluted femoral stem is a useful device in these patients. PMID- 17920479 TI - How precise is the determination of rotational alignment of the femoral prosthesis in total knee arthroplasty: an in vivo study. AB - Correct rotation of the femoral component is critical in avoiding patellar instability in total knee arthroplasty. This study attempted to compare the precision of 4 commonly used methods (transepicondylar axis [TEA], 3 degrees external rotation [ER] from posterior condylar line [PCL], perpendicular cut to Leo Whiteside line [WSL], and balanced flexion gap [GAP]) in determining the rotational alignment of the femoral prosthesis in an in vivo setting. The ranges of error were as follows: (1) TEA, 28 degrees (17 degrees internal rotation [IR] to 11 degrees ER); (2) PCL, 27 degrees (13 degrees IR to 14 degrees ER); (3) WSL, 32 degrees (17 degrees IR to 15 degrees ER); (4) GAP, 26 degrees (14 degrees IR to 12 degrees ER). If an error of more than 5 degrees from neutral alignment is defined as an outlier, 56% in TEA, 72% in PCL, 60% in WSL, and 20% in GAP methods would fall in the outlier zone. PMID- 17920478 TI - Total hip arthroplasty for bilaterally ankylosed hips. AB - The purpose of this study was to evaluate the results of bilateral total hip arthroplasties (THAs) with special regard to angle of cup position in patients with bilaterally ankylosed hips. Twenty-four ankylosed hips were converted to THAs in 12 patients. Their mean age was 36 years and they were followed up for more than 3 years (average, 11 years). The mean Harris hip score increased from 55.4 to 82.3 points. Osteolysis and loosening were found in 3 and 2 hips, respectively. Eleven cups of 24 hips (46%) were outside the safe ranges of Lewinnek. Overall outcome after bilateral conversion of bilaterally ankylosed hips to THA was favorable, but the fused position of the contralateral limb had the tendency to define the direction of the malposition of the acetabular cup. PMID- 17920480 TI - An additional reference axis for determining rotational alignment of the femoral component in total knee arthroplasty. AB - No studies have examined the trochlear line connecting the most anterior projections of the lateral and medial femoral condyles in relation to the surgical epicondylar axis. To determine if the trochlear line is more consistent relative to the transepicondylar axis than the posterior condylar axis and the Whiteside's line, the angles between the surgical epicondylar axis and each of the 3 axes in 50 knees of cadavers were measured using computed tomography scans. The results showed that the variability in the trochlear line for referencing the transepicondylar axis was comparable to those of the Whiteside line and the posterior condylar axis. The trochlear line may be considered as an additional reference axis for determining the rotational alignment of the femoral component in total knee arthroplasty. PMID- 17920481 TI - Comparison of robotic-assisted and conventional manual implantation of a primary total knee arthroplasty. AB - This study was aimed to compare robotic-assisted implantation of a total knee arthroplasty with conventional manual implantation. We controlled, randomized, and reviewed 72 patients for total knee arthroplasty assigned to undergo either conventional manual implantation (excluding navigation-assisted implantation cases) of a Zimmer LPS prosthesis (Zimmer, Warsaw, Ind) (30 patients: group 1) or robotic-assisted implantation of such a prosthesis (32 patients: group 2). The femoral flexion angle (gamma angle) and tibial angle (delta angle) in the lateral x-ray of group 1 were 4.19 +/- 3.28 degrees and 89.7 +/- 1.7 degrees, and those of group 2 were 0.17 +/- 0.65 degrees and 85.5 +/- 0.92 degrees. The major complications were from improper small skin incision during a constraint attempt of minimally invasive surgery and during bulk fixation frame pins insertion. Robotic-assisted technology had definite advantages in terms of preoperative planning, accuracy of the intraoperative procedure, and postoperative follow-up, especially in the femoral flexion angle (gamma angle) and tibial flexion angle (delta angle) in the lateral x-ray, and in the femoral flexion angle (alpha angle) in the anteroposterior x-ray. But a disadvantage was the high complication rate in early stage. PMID- 17920482 TI - Antibiotic-loaded cement articulating spacer for 2-stage reimplantation in infected total knee arthroplasty: a simple and economic method. AB - We presented a simple and economic method of preparing articulating antibiotic loaded cement spacers for treatment of infection after total knee arthroplasty. From 1996 to 2004, 28 infected total knee arthroplasties were treated with 2 stage reimplantation. Static spacers were used in 7 knees, and articulating spacers were used in 21 knees. A minimum of 2 years' follow-up after final treatment was evaluated. In the static group, 1 (14%) knee had recurrence of infection. In the articulating group, 2 (9%) knees had recurrence of infection with the original organism. Patients receiving articulating spacer had better range of motion, better knee score, and less bone loss than patients with static spacer. PMID- 17920483 TI - Histologic findings at 14 and 18 years after cemented total hip arthroplasty with interface bioactive bone cement technique. AB - To augment cement-bone bonding in total hip arthroplasty, we interposed hydroxyapatite (HA) granules at the interface. We report the histologic findings of 2 cases 14 and 18 years after cemented total hip arthroplasty with this technique. We found that a thick bony layer was observed where rich layers of HA granules existed. In areas without HA granules, the trabeculae did not form a thick bony layer. These bony layers around HA granules were observed at the interface of non-weight-bearing and weight-bearing areas in 2 acetabula after long-term follow-up. PMID- 17920484 TI - Osteoarthritis knee with patellar fracture in the elderly: single-stage fixation and total knee arthroplasty. AB - Periarticular fractures that occur in a previously osteoarthritic knee, especially in an elderly patient, will be associated with other comorbidities and is a treatment challenge. Early mobilization is a key treatment principle in the geriatric population. We present a case report of an elderly diabetic patient with osteoarthritis of the knee joint who sustained a patellar fracture and was treated with single-stage primary total knee arthroplasty and fixation of the patellar fracture. PMID- 17920485 TI - Computed tomography-based navigation to determine the socket location in total hip arthroplasty of an osteoarthritis hip with a large leg length discrepancy due to severe acetabular dysplasia. AB - For osteoarthritis hips due to severe acetabular dysplasia such as Crowe type 3 or 4, placement of the socket is a difficult procedure in total hip arthroplasty. Because the acetabular bone stock is poor, suitable location for the socket is very limited with respect to achieving good coverage with the host bone. A 51 year-old woman who had an osteoarthritis hip with a large leg length discrepancy due to severe acetabular dysplasia required total hip arthroplasty. The purpose of the total hip arthroplasty was to improve the hip disorder as well as to reduce the leg length discrepancy to achieve good gait function. We present technical solutions to aid the surgeons in placing the acetabular socket at the proper location by using computed tomography-based navigation system. PMID- 17920487 TI - The long-awaited Cochrane review of 2-phase treatment. PMID- 17920488 TI - Why do nickel-titanium archwires fracture? PMID- 17920490 TI - Need for training sessions for orthodontists in the use of the American Board of Orthodontics objective grading system. AB - INTRODUCTION: In recent years, the importance of quantitative assessment of orthodontic treatment with measures such as the objective grading system (OGS) has been emphasized for evaluating clinical outcomes in universities and clinics. Furthermore, quantitative evaluation is essential for passing orthodontic boards and examinations. However, countries outside the United States, the methods and the necessity of training sessions in the use of the OGS have not been evaluated. The OGS has great potential to be the "gold standard" to evaluate finished patients all around the world. Students and faculty members of Okayama University in Japan examined the necessity of training and the efficacy of 2 training methods. METHODS: Twenty-six orthodontic faculty members and graduate students participated. Three scoring and 2 training sessions were held by using 3 sets of cast models distributed by the American Board of Orthodontics and 6 sets of cast models prepared for this study. Statistical comparisons were done before and after the training sessions by analysis of variance and independent t test. RESULTS: Before the training session, the average difference in the total OGS scores for the 3 cases was 15.8 +/- 0.2 points. Significant reductions in the difference in the OGS score (7.3 +/- 2.4) were observed after 1 training session. There was no significant difference after the first and second training sessions. However, there was a significant difference in the variability of the OGS scores between the first and second sessions. There was no significant difference between the 2 training methods. Among 8 scoring criteria, the greatest difference in OGS scores was observed in buccolingual inclination throughout the training sessions. Furthermore, cases with unacceptable occlusion (high OGS score) tended to be difficult to score compared with acceptable occlusion (low OGS score). CONCLUSIONS: The OGS was suggested as an ideal scoring index to quantitatively assess orthodontic treatment outcome without specific training with well-trained orthodontists. Among the 8 categories of the OGS, extra care might be needed when scoring the buccolingual inclination. However, 1 training session greatly improved the scoring reliability of orthodontic treatment outcomes with the OGS. PMID- 17920491 TI - Bone formation and tooth movement are synergistically enhanced by administration of EP4 agonist. AB - INTRODUCTION: Stimulation of the prostaglandin receptor EP4 induces bone anabolic responses to external loading. The aim of this study was to determine the possibility of synergistic enhancement of tooth movement along with bone formation in vivo through the stimulation of the prostaglandin receptor EP4. METHODS: The sample consisted of 25 male rats, 6 to 7 weeks old. Experimental tooth movement was induced in the maxillary right molars with a split-mouth design. During the 7-day experimental period, some rats were injected periodically with a specific EP4 agonist (ONO-AE1-329), a drug that binds to the EP4 receptor to mimic the actions of ligand binding; the injection site was on the tension side in the interproximal region between the first and second molars. Other rats received vehicle injection (saline solution only). For control, EP4 agonist or vehicle was injected into some rats, but tooth movement was not initiated. The amounts of tooth movement and bone volume of the paradental region were evaluated by using soft x-ray and micro-computed tomography. RESULTS: The injection of EP4 agonist alone did not induce notable changes in body weight, macro-structures, or bone volume compared with the vehicle-treated group or the untouched control side. EP4 agonist injection in conjunction with tooth movement significantly enhanced the amounts of tooth movement and regional bone volume in the tension side compared with the vehicle-treated group (P <.05). CONCLUSIONS: Our preliminary study suggests the possibilities of synergistic enhancement of the rates of tooth movement and bone formation during tooth movement through the local injection of a specific EP4 receptor agonist. However, more study on the synergistic effect on tooth movement and bone formation in response to an EP4 agonist is needed to determine its mechanisms. PMID- 17920492 TI - Cervical column morphology in patients with skeletal Class III malocclusion and mandibular overjet. AB - INTRODUCTION: The cervical column morphology of adults with skeletal Class III malocclusion and mandibular overjet was examined and compared with the cervical column morphology of a control group with neutral occlusion and normal craniofacial morphology. METHODS: The mandibular overjet group consisted of 57 patients (32 women, 25 men). The control group consisted of 21 subjects (15 women, 6 men). A visual assessment of the cervical column and measurements of the craniofacial dimensions were performed on a profile radiograph of each subject. RESULTS: In the mandibular overjet group, 61.4% had cervical column fusion, and 12.3% had posterior arch deficiency. Morphologic cervical column deviations occurred significantly more often in the mandibular overjet group than in the control group (P <.001). The most important craniofacial parameters for fusion of the cervical column were maxillary retrognathism (P <.05) and mandibular overjet (P <.05) (R2 = 0.22). The most important factor for posterior arch deficiency was mandibular inclination (P <.05) (R2 = 0.12). CONCLUSIONS: These results are considered an important first step in phenotypic differentiation of skeletal Class III malocclusions that can be associated with different genotypes. This is important for the diagnosis and the correct treatment of these patients. PMID- 17920493 TI - Factors affecting buccal bone changes of maxillary posterior teeth after rapid maxillary expansion. AB - INTRODUCTION: The purpose of this study was to use cone-beam computed tomography (CBCT) images to determine the factors that might affect buccal bone changes of maxillary posterior teeth after rapid maxillary expansion (RME). METHODS: Thirty consecutive patients (17 boys, 13 girls; mean age, 13.8 +/- 1.7 years) who required RME as part of their orthodontic treatment and had the pre-RME (T1) and post-RME (T2) CBCT images available were included in the study. The T1 and T2 measurements of interdental distance, interdental angle (IA), buccal bone thickness (BBT), and buccal marginal bone levels (BMBL) of the first premolar (P1), the second premolar (P2), and the first molar (M1) were compared with the Friedman and the Wilcoxon signed rank tests. To determine which variables were associated with the changes in IA, BBT, and BMBL, the Spearman rank correlation analysis was performed (alpha = .05). RESULTS AND CONCLUSIONS: The results suggest that buccal crown tipping, and reduction of BBT and BMBL of the maxillary posterior teeth are the expected immediate effects of RME. There were no significant differences in dental expansion among P1, P2, and M1 (P >.05). P2 had clinically more buccal crown tipping (P = .116) but statistically less reduction in BBT and BMBL (P <.0001 and P = .001) than P1 and M1. Buccal bone changes and dental tipping on P2 were not affected by any other variables. Factors that showed significant correlation to buccal bone changes and dental tipping on P1 and M1 were age, appliance expansion, initial buccal bone thickness, and differential expansion (P <.05), but rate of expansion and retention time had no significant association (P >.05). PMID- 17920494 TI - Influence of the quality of the finished occlusion on postretention occlusal relapse. AB - INTRODUCTION: In this study, we aimed to evaluate the influence of the quality of the finished occlusion on postretention occlusal stability. METHODS: The sample comprised 87 patients with Class I malocclusion, treated with extraction of the 4 first premolars and edgewise mechanics; they were divided into 2 groups, according to the quality of their finished occlusions. Group 1 included 44 subjects (23 boys, 21 girls) with posttreatment peer assessment rating (PAR) scores from 0 to 5. The mean pretreatment age was 13.74 years (SD 2.14). The mean treatment time was 1.92 years (SD 0.57), the mean retention time was 1.75 years (SD 0.96), and the mean time of posttreatment evaluation was 5.17 years (SD 1.82). Group 2 included 43 subjects (22 boys, 21 girls) with posttreatment PAR scores greater than 5. The mean initial age was 13.34 years (SD 1.35). The mean treatment time was 2.20 years (SD 0.66), the mean retention time was 1.77 years (SD 0.78), and the mean posttreatment evaluation was 5.47 years (SD 1.60). The PAR and the Little irregularity indexes were measured on the dental casts at pretreatment, posttreatment, and postretention. Intergroup comparisons were made with independent t tests, and the Pearson correlation coefficient was applied to the PAR score for the whole sample at the times evaluated. RESULTS: Well-finished patients had lower posttreatment and postretention PAR scores and greater changes during the treatment and posttreatment periods than did the poorly finished patients. For the Little irregularity index, the only difference between the groups was at the posttreatment stage; group 1 had a smaller irregularity score than group 2. The correlation coefficients showed that the greater the treatment changes, the smaller the posttreatment PAR score and the greater the relapse. But the higher the posttreatment PAR score, the higher the postretention PAR score. CONCLUSIONS: It was concluded that the greater the quality of the orthodontic finished occlusion, the greater are the treatment changes and the amount of relapse and the better is the occlusal status at the postretention stage in Class I malocclusion treated with 4 premolar extractions. PMID- 17920495 TI - Accuracy of cone-beam computed tomography imaging of the temporomandibular joint: comparisons with panoramic radiology and linear tomography. AB - INTRODUCTION: Cone-beam computed tomography (CBCT) is increasingly being used as an imaging modality, particularly in the assessment of the temporomandibular joint (TMJ). A blinded observational cross-sectional in-vitro study was conducted to compare the diagnostic accuracy of observers viewing images made with CBCT, panoramic radiography, and linear tomography. The task was to detect cortical erosions affecting the mandibular condylar head. METHODS: The sample consisted of 37 TMJ articulations from 30 skulls with either normal condylar morphology (n = 19) or erosion of the lateral pole (n = 18). The articulations were imaged by using corrected angle linear tomography (TOMO), normal (Pan-N) and TMJ-specific (Pan-TM) panoramic radiography, and CBCT. Digital images were obtained with photostimulable phosphor plates for all modalities except CBCT. The CBCT detector used an amorphous silicon flat-panel array combined with cesium iodide. Images and 10 rereads were presented to 10 observers on a flat-panel display at a pixel to-monitor ratio of 1:1. CBCT multi-planar images were presented both statically (CBCT-S) and interactively (CBCT-I). The observers were permitted to scroll through axial (0.4 mm) and para-sagittal (1 mm) sections and then independently rate their confidence about the presence or absence of cortical erosion. Intraobserver reliability was determined by weighted kappa and diagnostic accuracy by the fitted area under the ROC curve. Means were compared by using ANOVA (P < or =.05). RESULTS: Intraobserver reliability was moderate (0.57 +/- 0.22; range, 0.34-0.78). Pan-N (0.72 +/- 0.15), CBCT-I (0.65 +/- 0.21), and CBCT S (0.65 +/- 0.17) reliability was significantly greater than TOMO (0.44 +/- 0.25). The diagnostic accuracy of CBCT-I (0.95 +/- 0.05) and CBCT-S (0.77 +/- 0.17) was significantly greater than all other modalities (Pan-N [0.64 +/- 0.11], Pan-TM [0.55 +/- 0.11], TOMO [0.58 +/- 0.15]). CBCT-I was also more accurate than CBCT-S, and Pan-N was more accurate than Pan-TM and TOMO. CONCLUSIONS: CBCT images provide superior reliability and greater accuracy than TOMO and TMJ panoramic projections in the detection of condylar cortical erosion. PMID- 17920496 TI - Facial growth prediction: a comparison of methodologies. AB - INTRODUCTION: The purpose of this study was to compare 3 methodologies that predict facial growth relative to incremental and positional changes over short- and long-term time periods. The 3 prediction systems that were tested and compared were the Ricketts analysis, the Johnston grid analysis, and the Fishman maturational analysis. METHODS: Serial lateral cephalometric headfilms and hand wrist films of 50 untreated subjects from the Burlington Growth Center at the University of Toronto, Toronto, Ontario, Canada, were used. The sample was subdivided by skeletal classification and further subdivided by sex. The sample comprised 19 skeletal Class I subjects (10 female, 9 male), 20 Class II subjects (10 female, 10 male), and 11 Class III subjects (6 female, 5 male). Three serial headplates were traced on each subject, and the associated chronologic and maturational ages were noted. The subjects were divided into a young adolescent subgroup approximately 9 years of age (T1), a midadolescent subgroup approximately 13 years of age (T2), and a late adolescent-adult age group approximately 20 years of age (T3). Predictions were made between the time periods. To measure and compare the predicted amounts of downward and forward skeletal growth vs the actual growth values, anatomic landmarks were chosen in the cranial base and on the maxilla and the mandible (Point A and gnathion), and increases between the respective points were measured. A paired t test was applied to each test group for the statistical analysis. Graphic representations were also created in the form of "positional envelopes" that encapsulated the predictive data for each methodology. To evaluate both incremental and positional accuracy, these data envelopes were subsequently related to the true positions of Point A and gnathion. RESULTS AND CONCLUSIONS: Although not the case in every instance, it was demonstrated that the maturationally oriented Fishman analysis was superior to both the chronologically based Johnston grid and Ricketts analysis for short- and long-term predictions. PMID- 17920497 TI - Morphometric correlation between facial soft-tissue profile shape and skeletal pattern in children and adolescents. AB - INTRODUCTION: Prediction of soft-tissue outline shape from skeletal remains is useful in forensics and archaeology. The inverse problem, the assessment of underlying skeletal relationships from the external appearance, is pertinent in orthodontics. The purposes of this study were to assess the correlation between craniofacial shape and shape of the soft-tissue profile outline and to determine the extent to which it might be possible to predict the latter from the former. METHODS: Lateral cephalograms from 170 consecutive orthodontic patients were used, and 17 skeletal, 2 dental, and 22 soft-tissue landmarks were digitized and processed by using Procrustes superimposition and principal component analysis. The principal components of the skeletal and soft-tissue shapes were entered into the correlation analysis. RESULTS: Significant correlations were found between the skeletal and the soft-tissue components. The use of 7 anterior skeletal landmarks (not including the nasal bone) resulted in a predictive power (coefficient of determination) of 38% of the variability of soft-tissue shape. This increased to almost 50% by adding nasal and incisor points but showed only a slight further improvement by incorporating posterior skeletal landmarks. CONCLUSIONS: Anterior skeletal and dental landmarks can be used to predict soft tissue profile shape with a 50% power in children and adolescents. PMID- 17920498 TI - Use of conventional tomography to evaluate changes in the nasal cavity with rapid palatal expansion. AB - INTRODUCTION: The relationship between nasal airway resistance and the use of rapid palatal expansion appliances remains controversial. The purpose of this study was to use conventional tomography to determine the anatomical changes in the nasal cavity after maxillary expansion. METHODS: Nineteen patients (aged 8-15 years) were included in the study. Tomograms were taken before expansion (T1), immediately after expansion (T2), and 3 months after expansion (T3). Areas for the left and right anterior, middle, and posterior nasal cavity and total volume were calculated by using the computer software, AutoCAD LT 2005. Data were analyzed with paired t tests. RESULTS: Significant increases in area were found in the anterior nasal cavity from T1 to T2 (0.85 +/- 1.19 cm2, 11.7% increase), T2 to T3 (1.18 +/- 1.2 cm2, 22.2% increase), and T1 to T3 (2.6 +/- 1.7 cm2, 35.7% increase) (P <.05). Similar increases were found in the middle and posterior nasal cavity. Significant increases in volume were found from T1 to T2 (2.1 +/- 2.7 cm3, 10.7% increase), T2 to T3 (4.9 +/- 2.3 cm3, 22.6% increase), and T1 to T3 (6.99 +/- 2.45 cm3, 27.8% increase). No significant differences were found in the area or the volume of the left and right sides of the nasal cavity. Individual variations in response to maxillary expansion were large for most of the parameters tested. CONCLUSIONS: These data suggest that rapid palatal expansion is usually accompanied by increases in area and volume of the nasal cavity, and these changes remain stable 3 months after maxillary expansion. PMID- 17920499 TI - Facemask therapy with and without expansion. AB - INTRODUCTION: The aim of this study was to compare the effects of facemask treatment with and without expansion in patients with skeletal Class lll malocclusion. METHODS: The material for this study consisted of the pretreatment and posttreatment lateral cephalograms of 28 subjects with skeletal and dental Class lll malocclusions, divided into 2 groups of 14. In the facemask with expansion group (FMEXP; 8 girls, 6 boys; mean age, 11 years 1 month), each subject wore a Delaire-type facemask and a bonded rapid maxillary expansion appliance. In the facemask only group (FM; 8 girls, 6 boys; mean age, 11 years 6 months), each subject wore a Delaire-type facemask with a removable appliance. A third group of untreated children with Class lll malocclusion (7 girls, 7 boys; mean age, 10 years 2 months) was used as the control sample. RESULTS: The forward displacement of the maxilla in both treated groups was significantly greater than in the control group. The forward displacement of pogonion and increase in facial depth in the control group showed significant differences compared with the treatment groups. Extrusion of the maxillary molars was significantly greater in the FM group than in the control group. The increase in the mandibular plane angle and the decrease in the facial axis were significantly different between the FMEXP group and the control group. Molar relationship increased more in the FM group than in the FMEXP group; this was the only significant difference between the treatment groups. In both treatment groups, dental and skeletal treatment of subjects with Class lll malocclusion were achieved. CONCLUSIONS: If the treatment indications are appropriate, facemask therapy can be an effective method for Class III treatment. PMID- 17920500 TI - Photographic soft-tissue profile analysis in children at 6 years of age. AB - INTRODUCTION: Profile photographs can be a valuable, noninvasive tool for early orthodontic diagnosis and treatment planning. METHODS: Left-side profile photographs were obtained of 181 normal, healthy children at age 6 years. Standardized landmarks were digitized on the photographs, and several linear and angular measurements were computed. The children were divided according to dental class and sex. Comparisons were made by 2-way analyses of variance. RESULTS: Facial convexity (larger in boys than in girls), Sn-N-Sl, and nasolabial and interlabial angles differed significantly (P <.01) between the sexes. Girls had significantly less labial protrusion than boys. Facial height was significantly greater in children with dental Class II, without sex differences. All analyzed angles were significantly influenced by dental class. Facial convexity was smaller in children with dental Class II. Cutaneous class was larger, and lips were more prominent in children with dental Class II than in those with dental Class III. CONCLUSIONS: The significant relationship between dental and cutaneous classes has important implications for orthodontic diagnosis and treatment. Dental class can usefully represent facial esthetics, and orthodontic procedures that modify dental occlusion might cause important repercussions to facial soft tissues. PMID- 17920501 TI - Timing of Class II treatment: skeletal changes comparing 1-phase and 2-phase treatment. AB - INTRODUCTION: Previous studies reported small but significant skeletal changes as a result of early treatment of Class II malocclusion with headgear and functional appliances. In this study, we report on the skeletal changes for 1-phase and 2 phase treatment of Class II malocclusion. METHODS: This was a prospective randomized clinical trial conducted sy the Department of Orthodontics at the University of Florida between 1990 and 2000. A total of 261 subjects demonstrating at least a one half-cusp Class II molar relationship and meeting the inclusion criteria were enrolled in the study and had at least 1 follow-up visit. During phase 1, 86 subjects were treated with a bionator, 95 were treated with a headgear/biteplane, and 80 served as the observation group. For phase 2, all subjects were then treated with full orthodontics appliances. Skeletal changes were monitored with cephalograms taken at baseline, at the end of early Class II treatment or observation baseline, at the beginning of fixed appliances, and at end of orthodontic treatment. RESULTS: Overall skeletal changes at the end of phase 1 treatment were as follows: (1) SNA angle increased in the bionator (0.51) and the observation groups (0.67), whereas it decreased (-0.50) in the headgear/biteplane group; (2) SNB angle increased in the bionator (1.36) and the observation groups (0.84), whereas it remained unchanged (0.19) in the headgear/biteplane group; (3) ANB angle decreased in the bionator (-0.85) and the headgear/biteplane groups (-0.72), and was unchanged in the observation group; and (4) the mandibular plane angle increased (1.30) only in the headgear/biteplane group. By the end of full orthodontic treatment, the skeletal differences in all measurements for all 3 groups were within 1 degrees . Linear regression models showed that, during phase 1, baseline value and treatment group were significant. However, when the entire treatment period was considered, treatment group had no effect. CONCLUSIONS: There is temporary skeletal change as a result of phase I treatment with both appliances but no detectible skeletal difference between 1-phase and 2-phase treatment of Class II malocclusion by the end of full orthodontic treatment. PMID- 17920502 TI - Class II treatment efficiency in maxillary premolar extraction and nonextraction protocols. AB - INTRODUCTION: In this study, we compared the efficiency of nonextraction and 2 maxillary-premolar-extraction protocols in complete Class II malocclusion treatment. METHODS: A sample of 112 records from patients with complete Class II malocclusion was divided into 2 groups with the following characteristics: group 1, comprising 43 patients treated nonextraction with an initial mean age of 12.63 years; and group 2, comprising 69 patients treated with extraction of 2 maxillary premolars with an initial mean age of 13.91 years. To compare the efficiency of each treatment protocol, the initial and final occlusal statuses were evaluated on dental casts with the peer assessment rating (PAR) index, and the treatment time of each group was calculated from the clinical charts. Treatment efficiency was calculated by the ratio between the percentage of PAR reduction and the treatment time. PAR scores, treatment times, and treatment protocols' efficiencies of the groups were compared with the t test. RESULTS: The 2 maxillary-premolar-extraction protocol had a smaller final PAR score, a greater percentage of PAR reduction, and greater treatment efficiency than the nonextraction protocol of complete Class II malocclusion. CONCLUSION: The 2 maxillary-premolar-extraction protocol has greater treatment efficiency than the nonextraction protocol of complete Class II malocclusion. PMID- 17920503 TI - A long-term follow-up study of Class II malocclusion correction after treatment with Class II elastics or fixed functional appliances. AB - INTRODUCTION: The aim of this study was to compare dentofacial morphology and long-term follow-up changes in growing males with skeletal Class II malocclusions treated without extractions and with either Begg or Herbst appliances. METHODS: Lateral cephalograms were obtained at the start of treatment, after active treatment, and after long-term follow-up. RESULTS: Treatment effects differed between the groups, with usually more favorable effects in the Herbst group. However, during the follow-up period, many of the changes were reversed. During the total observation period, maxillary prognathism and lower anterior facial height increased more in the Begg group than in the Herbst group. CONCLUSIONS: Although there were initial marked differences in the treatment outcomes, most of these differences were not sustained over the longer term. PMID- 17920504 TI - Preoperative acetaminophen vs ibuprofen for control of pain after orthodontic separator placement. AB - INTRODUCTION: Pain control during orthodontics is an important aspect of patient compliance. The aim of this prospective, randomized, double-blind clinical trial was to compare the pain control effectiveness of acetaminophen (650 mg) with ibuprofen (400 mg) taken 1 hour before separator placement in adolescents. METHODS: The patients recorded their discomfort on a 100-mm visual analog scale during several activities (teeth not touching, chewing, and fitting back teeth together) and by selecting words adapted from the McGill Pain Questionnaire at 5 time intervals: immediately before separator placement, immediately after separator placement, 2 to 3 hours later, at bedtime, and on awakening the next morning. The patients (n = 33) were randomly assigned to the ibuprofen group or the acetaminophen group. A repeated-measures ANOVA was performed as a function of time and treatment group. RESULTS: Pain increased immediately after separator placement, lessened, and then increased to a peak the next morning. The most commonly selected words to describe pain were "annoying," "sore," and "tight." There was no significant difference in pain at any time after separator placement regardless of the medication taken. CONCLUSIONS: Acetaminophen and ibuprofen produced no significant differences in pain after separator placement. PMID- 17920505 TI - A randomized clinical trial comparing the efficacy of ibuprofen and paracetamol in the control of orthodontic pain. AB - INTRODUCTION: Previous research has shown that ibuprofen provides effective relief from orthodontic pain. The aim of this study was to ascertain whether paracetamol (also known as acetaminophen) provided pain relief of equivalent or greater magnitude. METHODS: A multicenter, noninferiority, randomized clinical trial was conducted in 3 orthodontic clinics; 159 patients aged 12 to 16 years attending for routine orthodontic treatment were randomly allocated to receive either 400 mg of oral ibuprofen or 1 g of oral paracetamol an hour before and again 6 hours after separator placement. Pain scores were recorded on 7 visual analog scales (10 cm) over a week. The margin of equivalence was defined as 10 mm. RESULTS: Mean orthodontic pain from 2 hours after separation to bedtime was 8.5 mm (90% CI: lower, 3.7; upper,13.2) higher in the paracetamol group. This confidence interval lies partly outside the margin of equivalence, suggesting that paracetamol is not equivalent, and excludes the value 0, suggesting that ibuprofen is superior. From day 1 onward, there was a trend for patients who had taken ibuprofen to experience less pain at most time intervals compared with the paracetamol group. Two doses of ibuprofen, taken on the day of separator placement, were insufficient to control orthodontic pain on day 1 after placement. CONCLUSIONS: A combination of preoperative and postoperative ibuprofen is more effective than paracetamol in the control of orthodontic pain. PMID- 17920506 TI - Light-emitting diode vs halogen light curing of orthodontic brackets: a 15-month clinical study of bond failures. AB - INTRODUCTION: Previous in-vitro investigations reported no significant differences in the bond strength of brackets cured with conventional halogen lamps and those cured with light-emitting diodes (LED), even though LED curing times are much shorter. However, it is not known how LED curing performs in the oral cavity. The purpose of this randomized clinical trial was to evaluate the clinical performance of brackets cured with 2 light-curing units: a conventional halogen unit and an LED. METHODS: Thirty patients treated with fixed appliances were included in this study. The bonding followed a contralateral quadrant pattern: in each patient, 2 quadrants were cured with the conventional halogen unit, and the other 2 quadrants were cured with the LED unit. The study had a single blind controlled design with a within-patient comparison of the 2 curing techniques, and the patients were allocated randomly. A total of 544 stainless steel brackets were examined for bracket failure. Location (tooth), cause, and date of failure were recorded over 15 months. Statistical analyses were performed with the Kaplan-Meier survival estimates and the log rank test. RESULTS: No statistically significant differences were found in total bond failure rate or in mean survival time between brackets cured with the halogen light and those cured with the LED. Neither were significant differences found between the 2 lights when the clinical performances of the maxillary and mandibular arches were compared, or when the posterior and anterior segments were compared. CONCLUSIONS: These results show that curing with an LED does not result in more bond failures or shorter time to failure when compared with conventional halogen light curing. Curing with an LED is an acceptable alternative to conventional halogen light curing. PMID- 17920507 TI - Flexural strengths of fiber-reinforced composites polymerized with conventional light-curing and additional postcuring. AB - INTRODUCTION: The purpose of this study was to evaluate the effect of hand light curing (Optilux 501; SDS Kerr, Danbury, Conn) and secondary oven polymerization (Liculite; Dentsply, Dreieich, Germany) on the mechanical properties of 2 sizes (diameters, 0.6 and 1.2 mm) of fiber-reinforced composites (FRCs) (Ever Stick; Stick Tech, Turku, Finland). METHODS: The FRC samples were divided into 8 groups. Each group consisted of 10 specimens. Each FRC sample was evaluated with a 3 point bending test with a universal testing machine. Groups 1, 3, 5, and 7 were hand light-cured with a halogen curing unit for 40 seconds. Groups 2, 4, 6, and 8 were light-cured with the same curing unit for 40 seconds, followed by polymerization for 15 minutes in a light-curing oven. Groups 1 through 4 were tested at 1-mm deflection, and groups 5 through 8 at 2-mm deflection. RESULTS: The results of ANOVA indicated significant differences among the various groups (P = .000). The post-hoc test showed that the 2-mm deflection groups had significantly higher flexural strengths than the 1-mm deflection groups (P = .000). Moreover, the 1.2-mm FRCs showed significantly higher flexural strength than the 0.6-mm FRCs (P = .000). No significant differences (P >.05) were found between the hand light-cured and the oven-polymerized groups. CONCLUSIONS: This investigation demonstrated that oven postcuring does not increase the flexural strength values of 0.6- and 1.2-mm FRCs compared with conventional hand light curing. Thus, hand light-curing of FRCs is recommended directly in the mouth for orthodontic purposes. PMID- 17920508 TI - Running loop in unusual molar extraction treatment. AB - Running loops have been used in mandibular second premolar extraction treatment instead of cherry or shoehorn closing loops in the Tweed technique. They can also be used when molar extraction patterns are unusual or if molars are missing or hopeless, or have extensive restorations. A running loop archwire is usually made of 0.018 x 0.025-in stainless steel, with a helical loop 5 mm mesial to the buccal tube. The effective tipback is usually about 20 degrees to 30 degrees to upright molars, and a slight toe-in is necessary to prevent mesiolingual rotation. As the extraction space closes, the distance between the running loop and the buccal tube is shortened. Then a helical loop can be moved mesially to maintain the distance and the force between them by simple archwire manipulation. Consequently, the extraction space can be closed with 1 running loop archwire after initial leveling. PMID- 17920509 TI - Dilemmas in treating a patient with severe bialveolar protrusion and a hyperdynamic lip. AB - A black woman with severe bialveolar protrusion and a hyperdynamic lip on smiling was treated with molar and premolar extractions and orthognathic surgery, including a maxillary 3-piece LeFort, a maxillary segmental osteotomy, a mandibular segmental osteotomy, and a midsymphyseal ostectomy of the mandible. PMID- 17920510 TI - Accuracy of linear measurements from imaging plate and lateral cephalometric images derived from cone-beam computed tomography. AB - INTRODUCTION: As orthodontic practice moves toward 3-dimensional cephalometric analyses, a solution is required to ensure sustained availability of well established projected treatment outcomes based on 2-dimensional analyses. This ex vivo study was conducted to compare the accuracy of linear measurements made on photostimulable phosphor cephalograms with 3 methods for simulating lateral cephalograms with cone-beam computed tomography (CBCT). METHODS: The linear distances between anatomical landmarks on dentate dry human skulls were measured by observers using digital calipers for S-N, Ba-N, M-N, ANS-N, ANS-PNS, Pog-Go, Go-M, Po-Or, and Go-Co. The skulls were imaged with CBCT with a single 360 degrees rotation, producing 306 basis images and achieving 0.4 mm isotrophic voxel resolution on volumetric reconstruction for making ray-sum reconstructed cephalograms. Two other cephalogram approaches were used with the CBCT system--a single transmission image generated as a scout image designed to check patient positioning before CBCT, and a single-frame lateral basis image. Conventional digital lateral cephalograms (LCs) were acquired with the photostimulable phosphor system. Images were imported into a cephalometric analysis program (Dolphin Imaging Cephalometric and Tracing Software, Chatsworth, Calif) to compute the included linear measurements. Analyses were repeated 3 times and statistically compared with measured anatomic truth with ANOVA (P < or =.05). The intraclass correlation coefficient was determined as an index of intra- and interobserver reliability. RESULTS: The intraclass correlation coefficient for the LCs was significantly less than for the measured anatomic truth and for all CBCT-derived images. CBCT images either produced with individual frames or reconstructed from the volumetric data set were accurate for all measurements except Pog-Go and Go-M. CBCT scout images had the second highest accuracy for all measurements except Pog-Go, Go-M, and Go-Co. Conventional LCs had the least accuracy; they were accurate only for Po-Or and ANS-N. CONCLUSIONS: CBCT-derived 2-dimensional LCs proved to be more accurate than LCs for most linear measurements calculated in the sagittal plane. No advantage was found over single frame basis images in using ray-sum generated cephalograms from the CBCT volumetric data set. PMID- 17920513 TI - Measuring the secretion of heat shock proteins from cells. AB - Heat shock proteins have been shown to be secreted from a number of cell types. Necrotic cells release heat shock proteins in a passive manner, whereas we, and others, have shown that viable cells secrete Hsp70 and Hsp60 through an active mechanism involving lysosomal vesicles and lipid rafts. This release of Hsp70 and Hsp60 is regulated, for example by being increased by elevated temperature. This article outlines procedures, using Hsp70 as the example, to: ensure the status of cells (viable, apoptotic or necrotic); identify the heat shock protein secreted; and quantify the secreted protein. Hsp70 has previously been quantified by ELISA, but newer methods are now being adopted, such as BIAcore and bead-based assays for use by FACS. These methods have the advantages of being more sensitive and requiring less sample than ELISA. The BIAcore has the potential to analyse Hsp70 ligands and provide affinity constants. The FACS bead assay system can be used to run multiplex assays. PMID- 17920514 TI - Engineering secretable forms of chaperones for immune modulation and vaccine development. AB - Heat shock proteins are present in almost all intracellular compartments and serve by folding newly synthesized proteins, disassembling unstable proteins, and assisting in the transportation of proteins within the cell. Under certain circumstances they are also present on the cell surface, and can be shed or secreted into the extracellular environment. Although they possess many functional roles, their ability to stimulate innate and antigen-specific immunity have made them attractive candidates for vaccine development. Here, we review some of the approaches that have been used to genetically engineer molecular chaperones for their secretion from tumor cells or targeting them to the plasma membrane of such cells in order to promote anti-tumor responses. Treatment of tumor cells engineered to secrete or display chaperones may be of benefit, particularly in the area of cell-based vaccine development. PMID- 17920512 TI - Mechanisms for Hsp70 secretion: crossing membranes without a leader. AB - Heat shock protein 70 (Hsp70) is released from cells of many types and plays a significant signaling role, particularly in the inflammatory and immune responses. However, Hsp70 does not contain a consensus secretory signal and thus cannot traverse the plasma membrane by conventional mechanisms. However, Hsp70 can be released from cells by active mechanism that are independent of de novo Hsp70 synthesis or cell death. This pathway is similar to one utilized by the leaderless protein interleukin 1beta. Hsp70 release involves transit through an endolysosomal compartment and is inhibited by lysosomotropic compounds. In addition, the rate of Hsp70 secretion correlates well with the appearance of the lysosomal marker LAMP1 on the cell surface, further suggesting the role for endolysosomes. The entry of Hsp70 into this secretory compartment appears to involve the ABC-family transporter proteins. While the cell signals involved in triggering Hsp70 release through this lysosomal pathway are largely unknown, recent data suggest a regulatory role for extracellular ATP. These mechanisms are also shared by interleukin 1beta secretion. Following release it has been shown that Hsp70 binds to adjacent cells, suggesting that the secreted protein participates in paracrine or autocrine interactions with adjacent cell surfaces. Thus an outline is beginning to of the mechanisms of Hsp70 secretion. Much further study will be required to fully elucidate mechanisms involved in targeting Hsp70 towards the non-canonical secretion pathways and its regulation. PMID- 17920515 TI - Hsp72 release: mechanisms and methodologies. AB - To date there are two mechanisms that are recognized by which heat shock proteins (HSP) are released from cells; a passive release mechanism, including necrotic cell death, severe blunt trauma, surgery and following infection with lytic viruses, and an active release mechanism which involves the non classical protein release pathway in which HSPs are released within highly immunologically potent exosomes and as free HSP. This chapter describes the experimental procedures that have been developed to study the mechanism by which stress induces the release of HSP72 into the circulation and addresses the biological significance of circulating HSP72 to host defense against disease. PMID- 17920516 TI - Cell surface receptors for molecular chaperones. AB - Heat shock proteins are intracellular molecular chaperones. However, extracellular heat shock proteins have recently been shown to mediate a range of powerful effects in inflammatory cells, neuronal cells and immune cells. These effects are transmitted by a number of cell surface receptors including LRP/CD91, CD40, Toll-like receptors, Scavenger receptors and c-type Lectins. However, although extracellular heat shock proteins are products of at least five different gene superfamilies, similar receptor types often trigger their effects. We have assessed heat shock protein binding to the different receptor types with particular regard to its role in tumor immunology. Heat shock protein 70 released from dying tumor cells or injected as part of a vaccine induces a remarkable range of immune effects. This molecular chaperone induces powerful pro inflammatory signaling cascades leading to the activation of antigen presenting cells. In addition, heat shock protein 70 is able to transport antigenic peptides as cargo from the tumor cell cytoplasm across the membranes of antigen presenting cells and deliver them to major histocompatability class I molecules, a process known as "cross-presentation". The resulting major histocompatability class I peptide complexes are then displayed on the cell surface by antigen presenting cells, leading to activation of cytotoxic T lymphocytes and tumor cell killing. Understanding how heat shock protein-receptor binding orchestrates individual components of tumor immunity will permit enhanced design of molecular chaperone based immunotherapy. PMID- 17920517 TI - Macrophage scavenger receptors and host-derived ligands. AB - The scavenger receptors are a large family of molecules that are structurally diverse and have been implicated in a range of functions. They are expressed by myeloid cells, selected endothelial cells and some epithelial cells and recognise many different ligands, including microbial pathogens as well as endogenous and modified host-derived molecules. This review will focus on the eight classes of scavenger receptors (class A-H) in terms of their structure, expression and recognition of host-derived ligands. Scavenger receptors have been implicated in a range of physiological and pathological processes, such as atherosclerosis and Alzheimer's disease, and function in adhesion and tissue maintenance. More recently, some of the scavenger receptors have been shown to mediate binding and endocytosis of chaperone proteins, such as the heat shock proteins, thereby playing an important role in antigen cross-presentation. PMID- 17920518 TI - Lectin-like oxidized LDL receptor-1 as extracellular chaperone receptor: its versatile functions and human diseases. AB - Well-known coronary risk factors such as hyperlipidemia, hypertension, smoking, and diabetes are reported to induce the oxidative stress. Under the oxidative stress, low-density lipoprotein (LDL) is oxidatively modified in the vasculature, and formed oxidized LDL induces endothelial dysfunction, expression of adhesion molecules and apoptosis of vascular smooth muscle cells. It has become evident that these cellular responses induced by oxidized LDL are mediated by lectin-like oxidized LDL receptor-1 (LOX-1). LOX-1 was originally identified from cultured aortic endothelial cells as a receptor for oxidized LDL; however, recent investigations revealed that LOX-1 has diverse roles in the host-defense system and inflammatory responses, and it is involved in the pathogenesis of various diseases such as atherosclerosis-based cardiovascular diseases and septic shock. Beside oxidized LDL, LOX-1 recognizes multiple ligands including apoptotic cells, platelets, advanced glycation end products, bacteria, and heat shock proteins (HSPs). The HSPs function as a chaperone to affect protein folding of newly synthesized or denatured proteins. There are accumulating evidences that the HSPs released into the extracellular space have potent biological activities and it may work as a kind of cytokines. It is demonstrated that LOX-1 works as a receptor for HSP70, since it has high affinity for HSP70. The interaction of LOX 1 with HSP70 is involved in the cross-presentation of antigen. Given the potent and wide variety of biological activities, more understanding their interaction provides potential therapeutic strategy for various human diseases. PMID- 17920519 TI - The search for the chaperonin 60 receptors. AB - Chaperonin (Cpn)60 proteins have the ability to activate human and murine myeloid cells. There is contradictory evidence that the receptor for this protein is either similar to that of lipopolysaccharide--CD14 and one or other toll-like receptor (e.g. TLR4) or is some other, undidentified, receptor. In an attempt to directly identify the receptor for Mycobacterium tuberculosis Cpn60.1 we have used two approaches. The first is to use Cpn60.1 as an affinity ligand to pull out the receptor from lysates of the murine monocyte cell line RAW 264.7. The second is to crosslink Cpn60.1 to its receptor on RAW cells and isolate the complex by immunoprecipitation. These methods have worked for other receptors. Using affinity chromatography, 2D SDS-PAGE and peptide mass fingerprinting with MALDI-TOF MS it was found that a number of proteins had the ability to bind to Cpn60.1 on an affinity matrix. We identified five proteins, three of which were likely to be on the cell surface. One of these proteins, the endoplasmic reticulum molecular chaperone, BiP did bind to Cpn60.1 with low affinity. Protein crosslinking studies proved inadequate as insufficient protein could be isolated for mass spectrometric identification. Thus, it appears that Cpn60.1, like Hsp70, may bind to a number of cell surface proteins. BiP appears to be one of these receptor proteins but more work is needed to identify those responsible for signalling. Of interest, CD14 and TLR4 were not identified in this study as a receptor for Cpn60.1. PMID- 17920520 TI - Heat shock protein 70 (Hsp70): membrane location, export and immunological relevance. AB - Stress or heat shock proteins (HSPs) are remarkably conserved in all living organisms. Their expression is induced in response to a variety of physiological and environmental insults. In the cytosol these proteins play an essential role as molecular chaperones by assisting the correct folding of nascent and stress accumulated misfolded proteins, preventing protein aggregation, transport of proteins, and supporting antigen processing and presentation. Following stress, intracellularly located HSPs fulfill protective functions and thus prevent lethal damage. In contrast, membrane-bound or extracellularly located HSPs act as danger signals and elicit immune responses mediated either by the adaptive or innate immune system. Here, HSPs act as carriers for immunogenic peptides, induce cytokine release or provide recognition sites for natural killer (NK) cells. This article will discuss methods for the detection of membrane-bound and extracellular HSPs and methods for determining their immunological functions. PMID- 17920522 TI - Cognitive neuroscience of ownership and agency. PMID- 17920521 TI - A non-receptor-mediated mechanism for internalization of molecular chaperones. AB - The evolving realization that stress proteins, which have for many years been considered to be exclusively intracellular molecules under normal conditions, can be released from viable cells via a number of potential routes/pathways has prompted interest into their extracellular biology and intercellular signaling properties. That the stress proteins Hsp60, Hsp70 and gp96 can elicit both pro- and anti-inflammatory effects suggests that these molecules play a key role in the maintenance of immunological homeostasis, and a better understanding of the immunobiology of extracellular stress proteins might reveal new and more effective approaches for controlling and managing infectious disease, inflammatory disease and cancer. A number of cell surface receptors for stress proteins have been identified, and the intracellular consequences of these cell surface receptor-ligand interactions have been characterized. To date, studies into the intercellular signaling properties of stress proteins and their interactions with antigen presenting cells have focused on specific receptor mediated uptake, and have not considered the fact that such cells can also take up proteins via non-specific endocytosis/pinocytosis. Herein we present a methodological approach for assessing receptor-mediated and non-receptor-mediated uptake of gp96 by rat bone marrow-derived dendritic cells. PMID- 17920523 TI - Is minimal self preserved in schizophrenia? A subcomponents view. PMID- 17920524 TI - Neurophenomenology and the study of self-consciousness. PMID- 17920525 TI - Pacemaker cell and atrial function: unravelling how calcium initiates and regulates the heart beat and how ionic dysfunction, 'channelopathies' and other membrane remodelling contribute to atrial fibrillation. PMID- 17920526 TI - Dietary supplementation with laminarin, a fermentable marine beta (1-3) glucan, protects against hepatotoxicity induced by LPS in rat by modulating immune response in the hepatic tissue. AB - We tested the hypothesis that laminarin (LAM), a beta (1-3) polysaccharide extracted from brown algae, can modulate the response to a systemic inflammation. Male Wistar rats (n=7 per group) were fed a standard diet (control) or a diet supplemented with LAM for 25 days (5% during 4 days followed by 10% during 21 days). Thereafter, Escherichia coli lipopolysaccharides (LPS; 10 mg/kg i.p.) were injected and the animals were sacrificed 24 h after LPS challenge. The hypothermia, hyperglycemia and hypertriglyceridemia occurring early after LPS administration were less pronounced in LAM-treated rats than in controls. The increase in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) activities - reflecting hepatic alterations - was lessened after LPS injection in LAM-treated rats compared to control rats. LAM treatment decreased serum monocytes number, nitrite (NO2) and tumor necrosis factor-alpha (TNF-alpha). LAM also modulated intra-hepatic immune cells: it lowered the occurrence of peroxidase-positive cells (corresponding to monocytes/neutrophils) and, in contrast, it increased the number of ED2-positive cells, corresponding to resident hepatic macrophages, i.e. Kupffer cells. In conclusion, the hepatoprotective effect of marine beta (1-3) glucan during endotoxic shock may be linked to its immunomodulatory properties. We propose that both lower recruitment of inflammatory cells inside the liver tissue and lower secretion of inflammatory mediators play a role in the tissue protective effect of LAM. These effects could be due to a direct effect of beta-glucan on immune cells, or to an indirect effect through their dietary fibre properties (fermentation in the gut). PMID- 17920527 TI - Inhibitory effects of Stewartia koreana on osteoclast differentiation and bone resorption. AB - Osteoclasts are responsible for bone lysis in several bone diseases such as osteoporosis and rheumatoid arthritis. Natural products from plants have been invaluable source in discovery of compounds for new therapies. In this study, we screened plant products for potential application to therapy for bone loss using a primary osteoclastogenesis culture system and found that extract of Stewartia koreana (SKE) had a strong inhibitory effect on osteoclast formation. To gain molecular insights, we examined the effect of SKE on signaling pathways and transcription factors stimulated by the osteoclast differentiation factor RANKL. SKE suppressed the induction of c-Fos and NFATc1 by RANKL. However, SKE did not inhibit NF-kappaB activation by RANKL. Among the MAPKs stimulated by RANKL, SKE significantly reduced the activation of ERK and p38. Therefore, the anti osteoclastogenic effect of SKE is likely to be elicited by interference with RANKL signaling to ERK and p38, which mediate the induction of c-Fos and subsequently that of NFATc1. Consistent with the in vitro effect on osteoclast differentiation, SKE showed a great inhibitory effect on in vivo bone loss in LPS challenged mice. Taken together, we demonstrated that SKE has inhibitory effects on osteoclast differentiation in vitro and confirmed its in vivo efficacy in prevention of inflammatory bone loss. PMID- 17920528 TI - Effects of lactose-beta-sitosterol and beta-sitosterol on ovalbumin-induced lung inflammation in actively sensitized mice. AB - Asthma is a disease marked by chronic lung inflammation and the number of patients suffering from asthma increases annually. Both beta-sitosterol (BS) and beta-sitosterol glucoside exist in a variety of plants and have anti-tumor, anti microbial, and immunomodulatory activities. However, the precise role of BS and beta-sitosterol glucoside in asthma has not been well understood. The aim of this study was to investigate the inhibitory effects of BS and lactose-BS (L-BS) on the pathophysiological process in ovalbumin-induced asthmatic mice. The total cells and eosinophils in the bronchoalveolar lavage (BAL) fluid markedly decreased (p<0.05) after L-BS or BS administration (1 mg/kg; i.p.), and the ROS production also decreased in comparison to the asthma control. Histopathological features were detected by performing histochemistry, including H&E and alcian blue & P.A.S staining. Both L-BS and BS mitigated the inflammation by eosinophil infiltration and mucus hypersecretion by goblet hyperplasia. These effects of L BS were superior to those of BS. L-BS and BS inhibited the increased mRNA and protein expression of IL-4 and IL-5 in the lung tissue and BAL fluid, respectively. The IgE concentration in the BAL fluid and serum was measured by performing ELISA and the ovalbumin-specific IgE in the BAL fluid was uniquely inhibited by L-BS (p<0.05). The splenocytes were isolated from the normal and asthmatic mice and incubated in the absence and presence of 100 microg/ml ovalbumin, respectively. L-BS blocked the survival rate of the splenocytes of the mice (p<0.01). This finding indicates the possibility of L-BS and BS as potential therapeutic molecules in asthma and may contribute to the need to improve current therapeutic drugs. PMID- 17920529 TI - Species selectivity of small-molecular antagonists for the CCR5 chemokine receptor. AB - The species selectivity of four structurally different compounds, SCH-351125, E 913, TAK-779 and UK-427857 has been examined using cloned human, rhesus, and mouse CCR5 receptors. SCH-351125 and E-913 potently inhibited the binding of [125I]-CCL3 to human CCR5, but their inhibitory activities against rhesus CCR5 were more than 10-fold weaker. In contrast, TAK-779 and UK-427857 inhibited binding to human and rhesus CCR5 with similar potency. The inhibitory activities of all four compounds against mice CCR5 receptors were weak. The inhibitory activities of the CCR5 antagonists in the [125I]-CCL3 binding assay agreed well with those induced by CCL3 in the intracellular calcium ([Ca(2+)]i) elevation assay. Mutational analysis of the human CCR5 receptor showed that its Ile198 component plays a critical role in the inhibitory activities of both SCH-351125 and E-913, but not that of TAK-779 or UK-427857. These results provide a structural basis for understanding how specific antagonists interact with CCR5, and will aid the process of creating new, improved CCR5 antagonists. PMID- 17920530 TI - Anti-brain cholinergic auto antibodies from primary Sjogren syndrome sera modify simultaneously cerebral nitric oxide and prostaglandin biosynthesis. AB - The presence of circulating antibodies from primary Sjogren Syndrome (pSS) patients enable to interact with rat cerebral frontal cortex by activating muscarinic acetylcholine receptors (mAChR). ELISA assay for PGE2 generation, nitric oxide synthase (NOS) activity was measured in cerebral frontal cortex slices by production of [U-14C]-citruline and mRNA isolation/quantitative PCR for COX-1 and COX-2 gene expression were carried out. By ELISA assay, it was shown that IgG from pSS patients reacted to cerebral frontal cortex cell surface and with human M1 and M3 mAChR. Beside pSS IgG displayed an agonistic-like activity stimulating NOS activity and PGE2 production associated with an increased COX-1 mRNA gene expression, without affecting COX-2 mRNA levels. Inhibition of phospholipase A2 (PLA2) and NOS prevented pSS IgG effects upon both PGE2 production and COX-1 mRNA levels. The results support the notion that serum IgG auto antibodies in pSS patients target cerebral mAChR may have pathogenic role in immune neuroinflammation and on cognitive dysfunction present in pSS patients. PMID- 17920531 TI - The serum amyloid A response to sterile silver nitrate in mice and its inhibition by dexamethasone and macrolide antibiotics. AB - Serum amyloid A protein (SAA) is an acute phase protein, known to be a sensitive indicator of inflammation. We have characterized the time course of the SAA response and inflammatory reaction to silver nitrate injection s.c. in mice and studied the effects of dexamethasone and macrolide antibiotics. 2% Sterile silver nitrate solution was injected s.c. into female BALB/c mice and blood collected by capillary action from the tail vein of each mouse at different time points. Hematological variables were determined, albumin by spectrophotometry and SAA and cytokines by ELISA. Animals were treated with either a single i.p. dose of dexamethasone (5-30 mg/kg) 1 h after or daily oral doses of macrolide antibiotics for 3 days. SAA concentrations after silver nitrate injection peaked at 24 h, preceded by increases in serum IL-1 beta and IL-6, associated with decreases in blood leukocytes and local tissue inflammation. Single dexamethasone treatment and daily dosing for 3 days with azithromycin, clarithromycin and roxithromycin (20-80 mg/kg p.o.), but not erythromycin (100-150 mg/kg p.o.), inhibited the increase in SAA but with varying time courses. SAA, measured continuously, is a useful marker of sterile inflammation in mice and is differentially inhibited by macrolide antibiotics. PMID- 17920532 TI - Sinomenine inhibits activation of rat retinal microglia induced by advanced glycation end products. AB - Diabetic retinopathy involves an inflammatory response in the retina characterized by an increase in inflammatory cytokines and activation of microglia. The degree of microglia activation may influence the extent of retina injury following an inflammatory stimulus. Cytokines, released by activated microglia, regulate the influx of inflammatory cells to the damaged area. Thus, therapeutic strategy to reduce cytokine expression in microglia would be neuroprotective. Sinomenine, an alkaloid isolated from the stem and root of Sinomenium acutum, has long been recognized as an anti-inflammatory drug for rheumatoid arthritis and also inhibits macrophage activation. In this study, we activated retinal microglia in culture with advanced glycation end products (AGEs) treatment and attempted to determine whether sinomenine could reduce the production of cytokines from the activated microglia at both gene and protein levels. Changes in inflammatory cytokines, TNF alpha, IL-1 beta and IL-6, were measured by semi-quantitative RT-PCR and enzyme-linked immunosorbent assay (ELISA) both in the presence and absence of AGEs. The effect of sinomenine on levels of reactive oxygen species (ROS) and the nuclear translocation of NF-kB p65 were studied with a laser confocal scanning microscope. AGEs treatment induced a significant release of TNF alpha, IL-1 beta, and IL-6 from retinal microglia. Sinomenine could inhibit release of these cytokines. Sinomenine attenuated ROS production in a dose-dependent fashion and reduced the nuclear translocation of NF-kB p65 in AGEs-activated retinal microglia in culture. PMID- 17920533 TI - Heme oxygenase-1 induction by (S)-enantiomer of YS-51 (YS-51S), a synthetic isoquinoline alkaloid, inhibits nitric oxide production and nuclear factor-kappaB translocation in ROS 17/2.8 cells activated with inflammatory stimulants. AB - Activation of the inducible nitric oxide synthase (iNOS) pathway contributes to inflammation-induced osteoporosis by suppressing bone formation and causing osteoblast apoptosis. We investigated the mechanism of action by which YS-51S, a synthetic isoquinoline alkaloid, inhibits iNOS expression and nitric oxide (NO) production in ROS 17/28 osteoblast cells activated with the mixture of TNF-alpha, IFN-gamma and LPS (MIX). YS-51S, concentration- and time-dependently, increased heme oxygenase (HO-1) expression. Treatment with YS-51S 1 h prior to MIX significantly reduced MIX-induced NO production and iNOS expression with the IC50 to NO production of 47+/-3.3 microM. Electrophoretic mobility shift assay (EMSA) and western blot analysis showed that YS-51S inhibited MIX-mediated activation and translocation of NF-kappaB to nucleus by suppressing the degradation of its inhibitory protein IkappaBalpha in cytoplasm. YS-51S also reduced NF-kappaB luciferase activity. In addition, an HO-1 inhibitor ZnPPIX, antagonized the inhibitory effect of YS-51S on iNOS expression and DNA strand break induced by MIX, indicating prevention of NO production by YS-51S is associated with HO-1 activity. Moreover, YS-51S inhibited the oxidation of cytochrome c(2+) by peroxynitrite (PN). Our results indicated that YS-51S may be beneficial in NO mediated inflammatory conditions such as rheumatoid arthritis by alleviating iNOS expression and NO-mediated cell death of osteoblast with 1) inducing HO-1 expression, 2) interfering the activation of NF-kappaB and 3) quenching of PN. PMID- 17920535 TI - Stimulatory effect of N-(1-deoxy-beta-d-fructopyranos-1-yl)-l-proline on antibody production in mice. AB - Synthesized N-(1-deoxy-beta-d-fructopyranos-1-yl)-l-proline (Fru-Pro), an Amadori rearrangement product, has been investigated for its immunomodulatory effects. We evaluated the effects of Fru-Pro on the in vivo and in vitro activity in the regulation of the mice immune response. Haemagglutination antibody titre, plaque forming cell assay, E-rosette forming assay and cytotoxic test were studied. It has been demonstrated that the compound shows immunostimulating properties. In conclusion, present investigation suggests that the Fru-Pro is biologically functional, improves the immune system and might be regarded as an immune response modulator. PMID- 17920534 TI - Bucillamine mechanism inhibiting IL-1beta-induced VEGF production from fibroblast like synoviocytes. AB - We investigated the bucillamine (Buc) mechanism inhibiting interleukin (IL)-1beta induced vascular endothelial growth factor (VEGF) production from human fibroblast-like synoviocytes (HFLS) which derived from the inflamed synovium of an RA patient using SA981, its active metabolite. HFLS did not produce IL-1beta, spontaneously. While SA981 partially inhibited IL-1beta-induced VEGF production at concentrations of 10 to 100 microM (10.1% and 14.2% inhibition of total VEGF production under IL-1beta coexistence condition, respectively), it failed to inhibit IL-1beta-induced IL-6 production at the same concentrations. IL-1beta induced phosphorylation of the mitogen-activated protein (MAP) kinases, IkappaBalpha, c-Jun and Akt. SA981 at a concentration of 100 microM partially inhibited IL-1beta-induced phosphorylation of p38MAPK and Akt (12.0% and 36.1% inhibition of each total amount of phosphoprotein under IL-1beta coexistence condition, respectively). The VEGF promoter includes four transcription factors: AP1, hypoxia-inducible factor (HIF), Sp1 and AP2 binding elements. HIF-1beta, Sp1 and AP1 increased under IL-1beta coexistence conditions. At a concentration of 100 microM, SA981 attenuated increases in HIF-1beta and Sp1 (10.1% and 19.8% inhibition of each total amount of transcription factor under IL-1beta coexistence condition, respectively), but not AP1. These results suggest that SA981 partially inhibits VEGF production via modifications on IL-1beta signaling. Attenuation of the expression of HIF-1beta and Sp1 (but not AP1) may be a key with respect to SA981's selective inhibition of VEGF production. PMID- 17920536 TI - Dietary supplementation with specific oligosaccharide mixtures decreases parameters of allergic asthma in mice. AB - Specific mixtures of prebiotic oligosaccharides showed immune modulatory effects in previous murine vaccination experiments, suggesting a shift towards T-helper 1 (Th1) immunity. These mixtures consisted of short-chain galacto-oligosaccharides (scGOS) and long-chain fructo-oligosaccharides (lcFOS) in a 9:1 ratio (Immunofortis), with or without pectin-derived acidic oligosaccharides (pAOS). To investigate whether these mixtures could suppress Th2-related responses, they were tested in an ovalbumin (OVA)-induced model for experimental allergic asthma in BALB/c mice. Supplementation with two mixtures of scGOS/lcFOS and scGOS/lcFOS/pAOS at approximately 1% (w/w% net oligosaccharides) in the diet, starting two weeks before OVA sensitization and lasting until the end of the experiment, decreased of several parameters of allergic asthma. The OVA-induced airway inflammation and hyperresponsiveness was significantly suppressed by both mixtures. Moreover, OVA-specific IgE titers were decreased by more than 25%, although this effect was not significant. The effects of the oligosaccharide mixture with pAOS appeared to be more pronounced than the effects of the scGOS/lcFOS mixture without pAOS, but a direct comparison between the mixtures was not made. Overall, the results further support the hypothesis that specific mixtures of oligosaccharides modulate the Th1/Th2 balance by enhancing Th1 related and suppressing Th2-related parameters. PMID- 17920537 TI - Neuroimmunotherapeutics comes of age. PMID- 17920538 TI - A rose by any other name? The potential consequences of microglial heterogeneity during CNS health and disease. AB - Microglial activation and macrophage infiltration into the CNS are common features of CNS autoimmune disease and of chronic neurodegenerative diseases. Because these cells largely express an overlapping set of common macrophage markers, it has been difficult to separate their respective contributions to disease onset and progression. This problem is further confounded by the many types of macrophages that have been termed microglia. Several approaches, ranging from molecular profiling of isolated cells to the generation of irradiation chimeric rodent models, are now beginning to generate rudimentary definitions distinguishing the various types of microglia and macrophages found within the CNS and the potential roles that these cells may play in health and disease. PMID- 17920539 TI - Targeting MMPs in acute and chronic neurological conditions. AB - The matrix metalloproteinases (MMPs) are important enzymes that regulate developmental processes, maintain normal physiology in adulthood and have reparative roles at specific stages after an insult to the nervous system. Conversely, the concordant presence and significant upregulation of several MMP members in virtually all neurological conditions result in pathology. Thus, the MMPs have diverse functions, capable of mediating repair and recovery on the one hand and being involved in producing injury on the other. Therefore, targeting MMPs in neurological conditions has become a complicated challenge. This article highlights the beneficial roles of MMPs in normal and reparative processes within the nervous system and discusses the detriments of MMPs encountered in pathology. We review the availability of MMP inhibitors for clinical use and propose that an important consideration for these inhibitors is timing and duration of their use. With acute injuries where a massive upregulation of several MMPs are observed in the early periods after the insult, early and short-term use of broad spectrum MMP inhibitors would seem logical. In chronic conditions where recurrent insults to the CNS are accompanied by prolonged upregulation of MMPs, thereby necessitating the chronic use of medications, the beneficial effects of MMPs in repair may be compromised by the long-term application of MMP inhibitors. In this review we have used spinal cord injury and multiple sclerosis as examples of acute and chronic neurological conditions, respectively, and we consider the use of MMP inhibitors in these states. PMID- 17920540 TI - Chemokines and chemokine receptors in neurological disease: raise, retain, or reduce? AB - Chemokines and chemokine receptors comprise a large number of molecules implicated in a wide range of physiological and pathological functions. Numerous studies have demonstrated the roles of chemokines and chemokine receptors: 1) during development, by regulating hematopoiesis, cardiogenesis, and vascular and cerebellar development; 2) during tumor biology, by controlling cell proliferation, angiogenesis, and metastasis; and 3), especially during leukocyte migration, by acting on firm adhesion, locomotion, diapedesis, and chemotaxis. This review focuses on chemokine and chemokine receptor involvement in diverse neurological diseases and their therapeutic potentials. Because of its induction or upregulation during CNS pathologies, members of the chemokine system can be used as biological markers. CXCR4 and CXCL12, by the correlation between their expression and the glioblastoma tumor progression, could be a marker to grade this type of CNS tumor. CCR1, by virtue of specific expression in Abeta plaques, may be a marker for Alzheimer pathology. Downregulation of CCL2 in cerebrospinal fluid may be a candidate to characterize multiple sclerosis (MS), but needs additional investigation. Moreover, chemokines and chemokine receptors represent interesting therapeutic targets. Using chemokine receptor antagonists, several studies provided exciting findings for potential neurological disease treatment. Chemokine receptor antagonists reduce disease severity in animal models of MS. In glioblastoma, a CXCR4 antagonist (AMD3100) showed an inhibition of tumor growth. Inhibition of chemokine receptor signaling is not the only therapeutic strategy: for example, CXCR4-CXCL12 has anti-inflammatory properties and CX3CL1-CX3CR1 controls neurotoxicity. Thus, chemokine biology suggests several approaches for treating neurological disease. PMID- 17920543 TI - Corticosteroids for multiple sclerosis: II. Application for disease-modifying effects. AB - Physicians who treat multiple sclerosis (MS) face the challenge of patients exhibiting ongoing disease activity, including exacerbations, loss of functional capabilities, intellectual decline, and radiologic progression, despite being on a disease-modifying agent (DMA). After searching for factors that might at least in part explain these changes--such as nonadherent drug-taking behavior, or the presence of interferon-neutralizing antibodies--some providers may ultimately decide to switch the patient to another DMA. In most circumstances, patients likely derive only partial effects from these agents, even in the absence of compromising factors. Thus, a number of factors must be considered in order to intensify the treatment regimen in response to disease progression. In the context of an inadequate treatment response to a DMA, some clinicians will convert the patient to an alternative therapy, and others will instead use a second agent in combination with the first (the so-called platform agent). In the first of this two-part series, we explored the use of anti-inflammatory CS and ACTH to treat MS exacerbations. Although we underscored the limited availability of evidence-based studies to support specific regimens for this purpose, there is an even greater paucity of data to support the routine use of these agents in order to achieve chronic disease-modifying effects in those who continue to deteriorate clinically, radiographically, or both. Without doubt, a number of factors influence the formulation of combination treatment plan for MS. Nevertheless, we will focus on the rationale and practical schemes that can be considered for using corticosteroids (CS) (and perhaps even ACTH) in an attempt to modify various domains of ongoing disease activity. PMID- 17920542 TI - Corticosteroids for multiple sclerosis: I. Application for treating exacerbations. AB - Multiple sclerosis (MS) is an inflammatory demyelinating disorder characterized by a multiphasic course of neurological exacerbations, periods of clinical remission, and, in most patients, ultimately progressive deterioration of functional capabilities. The relapsing-remitting phase of the disease involves acute interruption in neurological functioning relating to areas of inflammation in discrete central-tract systems. The treatment of MS exacerbations with anti inflammatory agents such as corticosteroids and adrenocorticotropic hormone has represented an established practice throughout the neurology community. Although there is scientific rationale supporting application of these agents for this purpose, the broad diversity of approaches to using these drugs in clinical practice is a derivative of expert opinion and anecdotal experience. Ultimately, the treatment of MS-related exacerbations is part science, but mostly art. This review discusses the pharmacology of these agents, to better understand how they may act to mitigate attacks and to provide some practical formulations for how to use them in the clinic for the benefit of patients. PMID- 17920544 TI - Interferon-beta treatment for multiple sclerosis. AB - Multiple sclerosis (MS) is the leading nontraumatic cause of neurologic disability in young adults. Interferon-beta, approved for use in 1993, was the first treatment to modify the course and prognosis of the disease and remains a mainstay of MS treatment. Numerous large-scale clinical trials in early, active patient populations have established the clinical efficacy of interferon-beta in reducing relapses and delaying disability progression. Although its mechanism of action remains incompletely understood, a reduction in active lesions seen on magnetic resonance imaging implies primary anti-inflammatory properties, a mechanism supported by basic immunologic research. Variation in individual patient responsiveness to interferon-beta may be due to disease variability or differential induction of interferon-stimulated genes. The magnitude of the therapeutic effect appears to be similar among products, but the optimal dose, route, and frequency of administration of the drug remain uncertain. PMID- 17920541 TI - MRI in multiple sclerosis: what's inside the toolbox? AB - Magnetic resonance imaging (MRI) has played a central role in the diagnosis and management of multiple sclerosis (MS). In addition, MRI metrics have become key supportive outcome measures to explore drug efficacy in clinical trials. Conventional MRI measures have contributed to the understanding of MS pathophysiology at the macroscopic level yet have failed to provide a complete picture of underlying MS pathology. They also show relatively weak relationships to clinical status such as predictive strength for clinical progression. Advanced quantitative MRI measures such as magnetization transfer, spectroscopy, diffusion imaging, and relaxometry techniques are somewhat more specific and sensitive for underlying pathology. These measures are particularly useful in revealing diffuse damage in cerebral white and gray matter and therefore may help resolve the dissociation between clinical and conventional MRI findings. In this article, we provide an overview of the array of tools available with brain and spinal cord MRI technology as it is applied to MS. We review the most recent data regarding the role of conventional and advanced MRI techniques in the assessment of MS. We focus on the most relevant pathologic and clinical correlation studies relevant to these measures. PMID- 17920546 TI - Immunosuppressive agents in multiple sclerosis. AB - Immunosuppressive agents have been used in multiple sclerosis (MS) for decades. The approval of several immunomodulatory agents against MS beginning in the 1990s, whose putative mechanisms of action appeared "more MS-specific," curtailed the importance of immunosuppressants, which made them treatment options of second choice. However, with the recent approval of mitoxantrone for treatment of patients with active forms of relapsing-remitting or secondary progressive MS and with a number of oral immunosuppressive agents being assessed in phase II and III clinical trials, a "renaissance" of this type of agents is currently occurring. This review provides an outline of the most important clinical studies and discusses relevant side effects of the major immunosuppressants (i.e., mitoxantrone, azathioprine, cyclophosphamide, methotrexate, mycophenolate mofetil, cladribine, and sirolimus/temsirolimus). The current knowledge of the putative mechanisms of action of these compounds is discussed. PMID- 17920545 TI - Mechanism of action of glatiramer acetate in treatment of multiple sclerosis. AB - Glatiramer acetate (GA) (Copolymer-1, Copaxone, Teva, Israel, YEAK) is a polypeptide-based therapy approved for the treatment of relapsing-remitting multiple sclerosis. Most investigations have attributed the immunomodulatory effect of GAs to its capability to alter T-cell differentiation. Specifically, GA treatment is believed to promote development of Th2-polarized GA-reactive CD4(+) T-cells, which may dampen neighboring inflammation within the central nervous system. Recent reports indicate that the deficiency in CD4(+)CD25(+)FoxP3(+) regulatory T-cells in multiple sclerosis is restored by GA treatment. GA also exerts immunomodulatory activity on antigen presenting cells, which participate in innate immune responses. These new findings represent a plausible explanation for GA-mediated T-cell immune modulation and may provide useful insight for the development of new and more effective treatment options for multiple sclerosis. PMID- 17920547 TI - Antigen-specific therapy of multiple sclerosis: the long-sought magic bullet. AB - The adaptive immune response in multiple sclerosis (MS) targets various myelin proteins and even some inducible heat shock proteins. A few attempts have been made to tolerize relapsing-remitting patients with MS to either full-length myelin basic protein or to a key peptide epitope between residues 83-99. These trials have demonstrated that this approach may potentially provide benefit to patients with relapsing- remitting MS. However, manipulation of responses to myelin proteins can have deleterious effects. The immune response to myelin components is positioned at a key tipping point in the pathophysiology of the disease. Clarification of the key target antigens in MS, and better understanding of practical methods to attain tolerance to a wide variety of myelin and neuronal molecules will provide the basis for the ultimately successful antigen specific therapy. PMID- 17920548 TI - Modulating co-stimulation. AB - The modulation of co-stimulatory pathways represents a novel therapeutic strategy to regulate autoimmune diseases. Auto-reactive CD4+ T cells play a critical role in initiating the immune response leading to inflammation and autoimmune diseases. Blocking co-stimulatory signals prevents T-cell activation, thus diminishing autoimmune responses and possibly preventing the progression of autoimmune disease. Blockade of several co-stimulatory pathways has been investigated in animal models and has led to clinical trials testing specific blocking agents in humans. In this review we will describe the role of co stimulatory pathways, primarily the CD28-B7 pathway, in autoimmune diseases, and we will present in vivo and in vitro studies supporting the efficacy of co stimulation blockade in animal models of autoimmune disease. Finally, we will discuss the clinical therapeutic efficacy of blocking monoclonal antibodies in preventing or reducing auto-antigen driven T-cell activation in humans with particular attention to the CD28/B7 pathway. Inhibiting co-stimulatory molecule interactions by using monoclonal antibodies seems to be an original approach to regulate autoimmune diseases in humans. PMID- 17920549 TI - Emerging therapies for multiple sclerosis. AB - This review examines the mode of action, safety profile and clinical efficacy of some of the most promising new therapeutic strategies for multiple sclerosis. Autologous hematopoietic stem cell transplantation can regenerate a new and tolerant immune system and is a potentially effective rescue therapy in a subset of patients with aggressive forms of MS refractory to approved immunomodulatory and immunosuppressive agents. High-dose cyclophosphamide without stem cell support is suggested to induce prolonged remissions through similar immunological mechanisms with less toxicity. Fingolimod (FTY720) is a novel oral immunomodulating agent that acts through preventing lymphocyte recirculation from lymphoid organs. Monoclonal antibody therapy has provided scientists and clinicians the opportunity to rationally direct the therapeutic intervention against specific molecules. Targeting molecules of the immune system such as CD52 (alemtuzumab), CD25 (daclizumab), VLA-4 (natalizumab) and CD20 (rituximab) have resulted in potent immunomodulatory effects through sometimes unpredicted mechanisms. The potential of immunoglobulins to induce remyelination in the CNS is being investigated in an attempt to develop therapies promoting tissue repair and functional recovery. The evidence supporting the potential of these emerging immunotherapies suggests that strong progress is being made in the development of effective cures for multiple sclerosis. PMID- 17920550 TI - Statins--treatment option for central nervous system autoimmune disease? AB - Statins, inhibitors of the enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase, are well-established agents to lower cholesterol levels and prevent cardiovascular morbidity. Independent of their lipid-lowering properties, statins have been shown to exert pleiotropic immunomodulatory effects in various animal models of human autoimmune disease. In experimental autoimmune encephalomyelitis, a murine model for multiple sclerosis, statins prevented disease onset and even reversed paralysis when treatment was initiated after experimental autoimmune encephalomyelitis was fully established. Furthermore, well-tolerated oral statins were recently shown to exert synergistic benefit in experimental autoimmune encephalomyelitis in combination with existing agents for multiple sclerosis therapy. Based primarily on these encouraging results, statins are now being tested in clinical trials as a monotherapy for multiple sclerosis, as well as in combination with approved disease-modifying therapies. PMID- 17920551 TI - Prospects for neural stem cell-based therapies for neurological diseases. AB - Neural stem and progenitor cells have great potential for the treatment of neurological disorders. However, many obstacles remain to translate this field to the patient's bedside, including rationales for using neural stem cells in individual neurological disorders; the challenges of neural stem cell biology; and the caveats of current strategies of isolation and culturing neural precursors. Addressing these challenges is critical for the translation of neural stem cell biology to the clinic. Recent work using neural stem cells has yielded novel biologic concepts such as the importance of the reciprocal interaction between neural stem cells and the neurodegenerative environment. The prospect of using transplants of neural stem cells and progenitors to treat neurological diseases requires a better understanding of the molecular mechanisms of both neural stem cell behavior in experimental models and the intrinsic repair capacity of the injured brain. PMID- 17920553 TI - Alkaline methanolysis of lipid extracts extends shotgun lipidomics analyses to the low-abundance regime of cellular sphingolipids. AB - Sphingolipids that contain a sphingoid base are composed of hundreds to thousands of distinct compounds, many of which serve as lipid regulators of biological functions. The global analysis of the large number of low-abundance sphingolipid molecular species has been hampered in many cases by the sphingolipid molecular species being overwhelmed by the quantity of other classes of lipid (e.g., glycerophospholipid) molecular species present, thereby imposing severe restrictions on the dynamic range of their measurement using shotgun lipidomics. Herein, we developed a facile approach in which the sphingolipids of cellular extracts were dramatically enriched by direct alkaline methanolysis of lipid extracts followed by extraction to remove the large majority of other endogenous lipid classes. Through direct infusion of the resultant enriched solution, we identified and quantitated a variety of very-low-abundance sphingolipid classes (e.g., sphingosine, psychosine, and lysosphingomyelin) and molecular species (e.g., sphingomyelin) using electrospray ionization mass spectrometry (i.e., shotgun sphingolipidomics). Accordingly, through utilization of these facile enrichment techniques, direct penetrance into the sphingolipidomes has been greatly extended, facilitating new insights into their metabolism and signaling functions in biological systems. PMID- 17920554 TI - A spectrophotometric assay for histone deacetylase 8. AB - Inhibitors for the classical protein deacetylase enzymes have been actively pursued to develop the next generation of cancer therapy. Developing a novel convenient assay platform for the classical enzyme-catalyzed reactions could thus facilitate the drug discovery process. Based on our previous studies demonstrating the functional mimicry of N(epsilon)-thioacetyl-lysine for N(epsilon)-acetyl-lysine in the reaction catalyzed by the classical enzyme histone deacetylase 8 (HDAC8) on a peptide template derived from the C terminus of the human p53 tumor suppressor protein, we have developed a spectrophotometric HDAC8 assay via quantifying thioacetate produced from the enzymatic dethioacetylation with Ellman's reagent 5,5'-dithiobis(2-nitrobenzoate). We further demonstrated that this novel assay was selective for HDAC8 versus HDAC1 and 2 and for other classical protein deacetylase enzymes present in the HeLa nuclear extracts, thus making it potentially suitable not only for screening HDAC8-selective inhibitors but also for selectively assessing HDAC8 activity under (patho)physiological conditions. PMID- 17920552 TI - Immunology of neurological gene therapy: how T cells modulate viral vector mediated therapeutic transgene expression through immunological synapses. AB - Gene therapy has been shown to be a powerful new approach to the treatment of brain diseases. Brain neurodegenerations, brain tumors, inherited brain diseases, and autoimmune disorders are currently recognized as proper targets for gene therapeutics. Advances in the development of viral vectors (especially improvements in their immune profiles), the capacity to regulate transgene expression, and identification of appropriate therapeutic constructs have made the transition into clinical trials for gene therapy possible. One particular remaining challenge is the immune response that could be raised against either the viral vectors themselves or any regulatory or therapeutic transgenes. Because of the structure of brain immune responses, viral gene transfer into the brain can, under certain circumstances, be invisible to the systemic immune response and thus not generate a deleterious immune attack. If, however, the systemic immune system is primed against any vector antigen, the systemic immune response eliminates transgene expression and thus curtails the therapeutic efficacy of gene therapy. Mechanistic studies of brain immune responses indicate that the adaptive arm of the immune system may indeed be able to kill transduced cells. To move neurological gene therapy into the clinic in an effective and safe manner, these are the developments needed: novel viral vectors that either display a reduced capacity to stimulate an adaptive immune response or become invisible to the immune system after the delivery of the vector genome to the nucleus of transduced cells, and ways either to steer the immune response away from cytotoxic responses or to induce tolerance to gene therapy products. PMID- 17920555 TI - Spread of recombinant Autographa californica nucleopolyhedrovirus in various tissues of silkworm Bombyx mori determined by real-time PCR. AB - A cassette harboring luciferase reporter driven by Bombyx mori A3 promoter was transferred to the bacmid AcDeltaEGT to generate the recombinant virus AcNPVA3Luc (where Ac represents Autographa californica, NPV represents nucleopolyhedrovirus, and A3Luc represents the firefly luciferase reporter cassette driven by the A3 promoter). Recombinant baculovirus was injected into the hemocoele of newly ecdysed fifth instar larvae of the silkworm. The infection of virus in various silkworm tissues was determined by real-time PCR. The profile of viral infection showed that the copy number of recombinant AcNPV (rAcNPV) increased the fastest in the hemocyte, followed by the fat body, Malpighian tubule, middle gut, and silk gland. Detecting in nonpermissive strain silkworm showed that there was no significant difference in the entry of rAcNPV into all tested tissues. The difference in viral infection reflected mainly the big difference in replication of rAcNPV in various tissues of silkworm larvae. Real-time quantitative RT-PCR showed that it was due to the different expression of genes involved in viral DNA replication. PMID- 17920556 TI - [Hypertensive emergencies]. AB - Hypertensive urgencies are common clinical occurrences in hypertensive patients. The definition of hypertensive urgencies (target blood pression) were not consistent, but involved often target end-organ damage in emergencies. Epidemiology and physiopathology are briefly described. Treatment practices of hypertensive crisis were difficult because of the lack of evidence supporting the use of one therapeutic agent over another and its posology. PMID- 17920557 TI - [A case of chest trauma-induced acute myocardial infarction]. AB - The occurrence of an acute myocardial infarction (MI) after chest trauma is a rare complication. We report a case of a 58-year-old man presenting with an acute anterolateral MI secondary to blunt chest trauma. Coronary angiography revealed a non significant lesion of a first diagonal branch without any atherosclerosis lesion on coronary artery. He was conservatively managed and resulted in a good prognosis. Based on this case, we discuss the path physiologic mechanism of MI following chest trauma. PMID- 17920558 TI - [Electrical storm]. AB - Electrical storm is defined as repeated occurrence of severe ventricular arrhythmias requiring multiple cardioversions, two or more or three or more following different studies. The clinical aspect can sometimes be made of multiple, self aggravating, life threatening accesses. There are three main clinical circumstances of occurrence: in patients equipped with intracardiac defibrillators, during the acute phase of myocardial infarction and in Brugada syndrome. 10 to 15% of patients with cardiac defibrillators are subject to electrical storms in a period of two years. The causative arrhythmia is most often ventricular tachycardia than ventricular fibrillation, especially in secondary prevention and if the initial arrhythmias justifying the device was a ventricular tachycardia. Precipitaing factors are present in one third of cases, mainly acute heart failure, ionic disorders and arrhythmogenic drugs. Predictive factors are age, left ventricular ejection fraction<35% and renal insufficiency. Arrhythmia reduction is obtained by electrical shock in 50% of cases, antitachycardi stimulation in 30% and in 20% by association of the two. Treatment, after elimination of inappropriate shocks, is mainly based on beta blockers and amiodarone, class I antiarrhythmics, lidocaine or bretylium in some cases, and sedation pushed to general anesthesia in some cases. Radio-frequency ablation and even heart transplantation have been proposed in extreme cases. Quinidine has been proved efficient in cases of Brugada syndrome. PMID- 17920559 TI - [Multidetector computed tomography coronarography: preliminary experience in real life settings]. AB - Multidetector computed tomography coronarography (MDCT) is a promising tool, offering non invasive anatomic evaluation of coronary arteries. We relate in this article our impressions after our first two years experience. METHODS: Retrospective study of the 328 patients who underwent MDCT examination between January 2005 and December 2006 at our community hospital. Per patient comparative analysis of the anatomical findings versus percutaneous angiography (PCA) in 61 cases. RESULTS: Out of 328 patients 61 (18.5%) underwent PCA. In these patients we found a sensibility, specificity, positive predictive value and negative predictive value of 100%, 54%, 59% and 100% for MDCT. The main factor responsible for inconclusive vessel analysis was excessive calcification. CONCLUSION: In routine use, MDCT enables one to rule out significant stenosis non invasively, with high sensitivity in an unselected patient population. PMID- 17920560 TI - [Acute myocardial infarction in octogenarians treated by primary coronary angioplasty: in hospital outcome, social evolution and long-term mortality. About a forty-seven consecutive patients study]. AB - This study evaluated the hospital outcome of octogenarian patients treated by primary angioplasty for acute myocardial infarction less than 12 hours. The long term mortality and social outcome were evaluated, social outcome was based on IADL and IADL-E scales corresponding to physical autonomy and level of dependence. We included from 1999 to 2005 47 old patients of 83.5+/-3.2 years. The reperfusion of the obstructed coronary artery was obtained in 45 patients (95.7%) with TIMI 3 flow. Hospital mortality was 25.5% and 4.2% without hemodynamic complication. A multivariate analysis identified a Killip score>1 as predictive factor of hospital mortality (OR=8.9 and p=0.05). The long-term survival without death with a follow-up of 26.3+/-18.3 months was 82.2% at 24 months and 70.6% at 48 months according to the Kaplan-Meier method. In an multivariate analysis, hospitalization exit without aspirine was associated with a higher long term mortality (OR=34.8 and p=0.04). According to the evolution of scores of IADL and IADL-E scales, patients had an excellent autonomy and good capacities before the infarction and they kept them on the long term. Thus octogenarians with acute myocardial infarction treated by primary angioplasty have a high in hospital and long term mortality but the survivors preserve an excellent daily autonomy. PMID- 17920561 TI - Cholesterol and Clioquinol modulation of A beta(1-42) interaction with phospholipid bilayers and metals. AB - The beta-sheet plaques that are the most obvious pathological feature of Alzheimer's disease are composed of amyloid-beta peptides and are highly enriched in the metal ions Zn, Fe and Cu. The interaction of the full-length amyloid peptide, A beta(1-42), with phospholipid lipid bilayers was studied in the presence of the metal-chelating drug, Clioquinol (CQ). The effect of cholesterol and metal ions was also determined using solid-state 31P and 2H NMR. CQ modulated the effect of metal ions on the integrity of the bilayer and although CQ perturbed the phospholipid membrane, the bilayer integrity was maintained. Model membranes enriched in cholesterol were studied under conditions of peptide association and incorporation. Solid-state NMR showed that the bilayer integrity was preserved in cholesterol-enriched membranes in comparison to phosphatidylcholine-phosphatidylserine bilayers. Changes in peptide structure, consistent with an increase in beta-sheet, were observed using specifically 13C labelled A beta(1-42) by magic angle spinning NMR. Results using aligned phosphatidylcholine bilayers and completely 15N-labelled peptide indicated that the peptide aggregated. The results are consistent with oligomeric beta-sheet structured peptides only partially penetrating the bilayer and cholesterol reducing the membrane disruption. PMID- 17920562 TI - Tissue-specific deletion of c-Jun in the pancreas has limited effects on pancreas formation. AB - It is well known that activating protein-1 (AP-1) is involved in a variety of cellular functions such as proliferation, differentiation, apoptosis, and oncogenesis. AP-1 is a dimer complex consisting of different subunits, and c-Jun is known to be one of its major components. In addition, it has been shown that mice lacking c-Jun are embryonic lethal and that c-Jun is essential for liver and heart development. However, the role of c-Jun in the pancreas is not well known. The aim of this study was to examine the possible role of c-Jun in the pancreas. First, c-Jun was strongly expressed in pancreatic duct-like structures at an embryonic stage, while a lower level of expression was observed in some part of the adult pancreas, implying that c-Jun might play a role during pancreas development. Second, to address this point, we generated pancreas-specific c-Jun knock-out mice (Ptf1a-Cre; c-Jun(flox/flox) mice) by crossing Ptf1a-Cre knock-in mice with c-Jun floxed mice. Ptf1a is a pancreatic transcription factor and its expression is confined to pancreatic stem/progenitor cells, which give rise to all three types of pancreatic tissue: endocrine, exocrine, and duct. Contrary to our expectation, however, there was no morphological difference in the pancreas between Ptf1a-Cre; c-Jun(flox/flox) and control mice. In addition, there was no difference in body weight, pancreas weight, and the expression of various pancreas-related factors (insulin, glucagon, cytokeratin, and amylase) between the two groups. Furthermore, there was no difference in glucose tolerance between Ptf1a-Cre; c-Jun(flox/flox) and control mice. Taken together, although we cannot exclude the possibility that c-Jun ablation is compensated by some unknown factors, c-Jun appears to be dispensable for pancreas development at least after ptf1a gene promoter is activated. PMID- 17920563 TI - Cinnamaldehyde suppresses toll-like receptor 4 activation mediated through the inhibition of receptor oligomerization. AB - Toll-like receptors (TLRs) play a critical role in induction of innate immune and inflammatory responses by recognizing invading pathogens or non-microbial endogenous molecules. TLRs have two major downstream signaling pathways, MyD88- and TRIF-dependent pathways leading to the activation of NFkappaB and IRF3 and the expression of inflammatory mediators. Deregulation of TLR activation is known to be closely linked to the increased risk of many chronic diseases. Cinnamaldehyde (3-phenyl-2-propenal) has been reported to inhibit NFkappaB activation induced by pro-inflammatory stimuli and to exert anti-inflammatory and anti-bacterial effects. However, the underlying mechanism has not been clearly identified. Our results showed that cinnamaldehyde suppressed the activation of NFkappaB and IRF3 induced by LPS, a TLR4 agonist, leading to the decreased expression of target genes such as COX-2 and IFNbeta in macrophages (RAW264.7). Cinnamaldehyde did not inhibit the activation of NFkappaB or IRF3 induced by MyD88-dependent (MyD88, IKKbeta) or TRIF-dependent (TRIF, TBK1) downstream signaling components. However, oligomerization of TLR4 induced by LPS was suppressed by cinnamaldehyde resulting in the downregulation of NFkappaB activation. Further, cinnamaldehyde inhibited ligand-independent NFkappaB activation induced by constitutively active TLR4 or wild-type TLR4. Our results demonstrated that the molecular target of cinnamaldehyde in TLR4 signaling is oligomerization process of receptor, but not downstream signaling molecules suggesting a novel mechanism for anti-inflammatory activity of cinnamaldehyde. PMID- 17920564 TI - Panic disorder, social anxiety disorder, and a possible medical syndrome previously linked to chromosome 13. AB - BACKGROUND: Several studies have identified increased medical problems among individuals with panic disorder (PD). We previously found that specific conditions--interstitial cystitis (IC), mitral valve prolapse (MVP), migraines, and thyroid disorders--aggregated non-randomly among panic families (we called this the "PD syndrome") and that families with and without the syndrome were genetically distinguishable on chromosome 13. We present data from a new case control study that replicates and extends the syndrome phenotype clinically. METHODS: Probands with a definite diagnosis and family history of PD (n=219), social anxiety disorder (SAD; n=199), or both (n=173) and 102 control subjects with no personal/family history of anxiety were interviewed with the SADS-LA diagnostic instrument. Medical history was obtained via medical checklist and the family history screen; IC symptoms were assessed with criteria developed by the National Institute for Diabetes and Digestive and Kidney Diseases. Subjects and interviewers were unaware of the syndrome hypothesis; final best-estimate diagnoses were blind to syndrome data. RESULTS: Probands with PD or SAD, as compared with control subjects, were five or more times as likely to report IC symptoms and twice as likely to report MVP and migraines (other genitourinary and cardiovascular problems were not elevated). First-degree relatives of probands with PD or SAD were also at increased risk for IC, MVP, thyroid problems, and headaches, regardless of whether the proband reported the same condition. CONCLUSIONS: These findings are consistent with previous data supporting a PD syndrome and further suggest that this syndrome might include other anxiety disorders well. PMID- 17920566 TI - Daily oscillation of phospholipase C beta4 in the mouse suprachiasmatic nucleus. AB - An endogenous biological clock located in the hypothalamic suprachiasmatic nucleus (SCN) regulates the timing of an organism's physiology and behavior. A variety of receptors are found on SCN pacemaker cells which permit the clock mechanism to respond to extra- and intra-SCN chemical messengers. A subset of these receptors is coupled to G-proteins, which when bound, lead to the activation of a variety of intracellular signaling cascades. One common signaling pathway employs the phosphotidylinositol-specific phospholipase C enzyme to increase intracellular calcium levels. A specific isoform of this enzyme, phospholipase C beta4, is of particular interest to circadian biologists because in its absence, mice display a circadian phenotype. Moreover, it has been shown to be associated with receptor types that are involved in clock resetting. Despite compelling data that this enzyme could be a critical component of an intracellular signaling pathway in the SCN, no study to date has investigated the possible oscillation of phospholipase C in any mammalian tissue. In the present study, we analyzed the temporal variation in the number of phospholipase C beta4 immunoreactive cells in the SCN. Herein, we show that PLCbeta4 levels oscillate in the SCN of mice housed in a light:dark photoperiod. Protein levels reached a significant peak during the early night and a trough during the day. The oscillation was considerably damped in the SCN of mice housed in constant dark conditions indicating the cycle is photoperiod-dependent. These data are critical to understanding the temporal regulation of a variety of inputs to the mammalian central circadian clock. PMID- 17920565 TI - Involvement of kappa-opioid and endocannabinoid system on Salvinorin A-induced reward. AB - BACKGROUND: The recreational drug, Salvinorin A, derived from the plant of Salvia divinorum, is a potent and selective kappa-opioid receptor agonist. The abuse of selective k-agonists is a novel phenomenon, the mechanism of which is not fully understood. METHODS: We investigated salvinorin A given SC on the conditioned place preference (.05-160 microg/kg) and intracerebroventricular (ICV) self administration (.01-1 microg/infusion) paradigms, in Wistar rats. RESULTS: The present results demonstrate the rewarding effects of Salvinorin A in a range of doses between .1 and 40 microg/kg SC for conditioned place preference test and .1 .5 microg/infusion for ICV self-administration. Highest doses (160 microg/kg for conditioned place preference test and 1 microg/infusion for ICV self administration) were aversive. The rewarding effect was antagonized by intraperitoneal (IP) pretreatment with the cannabinoid CB(1) receptor antagonist, rimonabant [N-piperidino-5-(4-chlorophenyl)1-(2,4-dichloro phenyl)-4 methyl pyrazole 3-carboxamide] (1 mg/kg), and the kappa-opioid receptor antagonist, nor binaltorphimine (nor-BNI) (10 mg/kg). In the shell of nucleus accumbens, dopamine extracellular levels were increased after administration of salvinorin A (40 microg/kg SC), reaching a maximum value of about 150%. CONCLUSIONS: These data provide the demonstration of the rewarding effects of Salvinorin A through an interaction between kappa-opioid and (endo)cannabinoid system in rats. PMID- 17920567 TI - Hearing development and spiral ganglion neurite growth in VASP deficient mice. AB - Vasodilator-stimulated phosphoprotein (VASP) has been found to be involved in intracellular signalling pathways and to play an important role in the actin associated organization and formation of the cytoskeleton. Since differential VASP expression was noted in inner ear tissues, the present study was performed to investigate the hearing development in VASP deficient mice. Hearing development in VASP-/- mice and wild type animals was investigated by auditory brain stem (ABR) measurements. In addition, inner ear tissues of wild type animals were tested for VASP expression using PCR, Western blot analysis, in situ hybridisation, and immunohistochemistry. To compare spiral ganglion (SG) neurite growth, SG explants from VASP-/- and wild type mice were analyzed under cell culture conditions. The electroacoustical results of the present study indicate that VASP deficient mice present with a later onset of hearing during postnatal development compared to wild type animals. Transient VASP expression was detected in neonatal SG of wild type mice. Tissue culture experiments with SG explants from VASP-/- animals revealed significant alterations in SG neurite extension compared to wild types. The present findings suggest a role for VASP during neonatal development of the mammalian cochlea and allow speculation on a possible delayed innervation of cochlear hair cells due to changes in SG neurite growth in VASP-deficient mice. Temporary VASP deficits in the neonatal inner ear may be compensated by related proteins like MENA leading to a delayed but complete development of hearing function in VASP-/- animals. PMID- 17920568 TI - Neurotransmitter systems of commissural interneurons in the lumbar spinal cord of neonatal rats. AB - The circuits that generate rhythmic locomotor activities are located in the ventromedial area of the lumbar spinal cord and comprise commissural interneurons necessary for left-right alternation during walking movements. In this study we injected biotinylated dextran amine (BDA) into the ventromedial gray matter of the lumbar spinal cord of neonatal rats to label commissural interneurons. Anterogradely labeled axons arose from the site of injection, crossed the midline in the anterior commissure and arborized extensively in the contralateral ventral horn of the spinal cord. The presence of neurotransmitter systems in labeled axon terminals of commissural interneurons was investigated by using antibodies raised against specific transmitter-related proteins. Boutons potentially containing inhibitory amino acids were identified by applying glutamic acid decarboxylase (GAD65/67) and glycine transporter 2 antibodies. Out of 1146 BDA-labeled axon terminals, 663 boutons were assumed on this basis to be inhibitory; 76% of these terminals were immunoreactive for glycine transporter, 53% were immunoreactive for GAD and about 30% of inhibitory boutons might contain both inhibitory amino acids. Boutons potentially containing putative excitatory neurotransmitter were revealed with antibodies raised against vesicular glutamate transporters 1 and 2. Out of 590 BDA-labeled boutons about one fourth (158) were immunoreactive for glutamate transporters. These mammalian commissural interneurons are compared to the glycinergic commissural interneurons in the swimming CPGs of lamprey and the Xenopus tadpole. Our results show that commissural interneurons in the mammalian spinal cord form a heterogeneous group including glutamatergic excitatory and GABAergic and glycinergic inhibitory neurons. PMID- 17920569 TI - Ebselen attenuates oxidative DNA damage and enhances its repair activity in the thalamus after focal cortical infarction in hypertensive rats. AB - Oxidative DNA damage has been proposed to be a major contributor to focal cerebral ischemic injury. However, little is known about the role of oxidative DNA damage in remote damage secondary to the primary infarction. In the present study, we investigated oxidative damage within the ventroposterior nucleus (VPN) after distal middle cerebral artery occlusion (MCAO) in hypertensive rats. We also examined the possible protective effect of ebselen, one glutathione peroxidase mimic, on delayed degeneration in the VPN after distal MCAO. Neuronal damage in the ipsilateral VPN was examined by Nissl staining. Oxidative DNA damage and base repair enzyme activity were assessed by analyzing immunoreactivity of 8-hydroxy-2'-deoxyguanosine (8-ohdG) and 8-oxoguanine DNA glycosylase (OGG1), respectively. The number of intact neurons in the ipsilateral VPN decreased by 52% compared to the contralateral side in ischemia group 2 weeks after distal cerebral cortical infarction. The immunoreactivity of 8-ohdG significantly increased while OGG1 immunoreactivity significantly decreased in the ipsilateral VPN 2 weeks after distal cortical infarction (all p<0.01). Compared with vehicle treatment, ebselen significantly attenuated the neuron loss, ameliorated ischemia-induced increase in 8-ohdG level as well as decrease in OGG1 level within the ipsilateral VPN (all p<0.01). OGG1 was further demonstrated to mainly express in neurons. These findings strongly suggest that oxidative DNA damage may be involved in the delayed neuronal death in the VPN region following distal MCAO. Furthermore, ebselen protects against the delayed damage in the VPN when given at 24 h following distal MCAO. PMID- 17920570 TI - Systemic transplantation of human adipose stem cells attenuated cerebral inflammation and degeneration in a hemorrhagic stroke model. AB - Adipose-derived stem cells (ASCs) are readily accessible multipotent mesenchymal stem cells and are known to secrete multiple growth factors, and thereby to have cytoprotective effects in various injury models. In the present study, the authors investigated the neuroprotective effect of ASCs in an intracerebral hemorrhage (ICH) model. ICH was induced via the stereotaxic infusion of collagenase, and human ASCs (three million cells per animal) isolated from human fresh fat tissue, were intravenously administered at 24 h post-ICH induction. Acute brain inflammation markers, namely, cell numbers positively stained for terminal transferase dUTP nick end labeling (TUNEL), myeloperoxidase (MPO), or OX 42, and brain water content were checked at 3 days post-ICH. In addition, the authors quantified brain degeneration by measuring hemispheric atrophy and perihematomal glial thickness at 6 weeks post-ICH, and determined modified limb placing behavioral scores weekly over 5 weeks post-ICH. The results showed that brain water content, TUNEL+, and MPO+ cell numbers were significantly reduced in the ASC-transplanted rats. ASC transplantation attenuated neurological deficits from 4 to 5 weeks post-ICH, and reduced both the brain atrophy and the glial proliferation at 6 weeks. Transplanted ASCs were found to densely populate perihematomal areas at 6 weeks, and to express endothelial markers (von Willebrand factor and endothelial barrier antigen), but not neuronal or glial markers. In summary, ASCs transplantation in the ICH model reduced both acute cerebral inflammation and chronic brain degeneration, and promoted long-term functional recovery. PMID- 17920571 TI - Interaural intensity difference and ear advantage in listening to dichotic consonant-vowel syllable pairs. AB - The right-ear advantage (REA) is typically observed in verbal dichotic listening, indicating a left hemisphere superiority for speech processing. The REA could be thought of as a bottom-up, stimulus-driven laterality effect, caused by the preponderance of the contralateral neural fibers from the right ear to the auditory/speech processing areas in the left temporal lobe. The REA can, however, be modified by explicitly requiring the listeners to focus their attention alternatively on the left or right-ear stimuli, thus either countering or enhancing the bottom-up processes through top-down attentional control. In the present study, we manipulated the strength of the bottom-up REA by inducing an intensity difference between the right-ear and left-ear speech inputs in order to make the REA either weaker (left-ear input>right-ear input) or stronger (left-ear input1000-fold less potent than ASMSP at activating gerbil NK(1) receptors. The selective NK(3) receptor agonist senktide evokes fear-related gerbil foot tapping, an effect which probably involves indirect enhancement of NK(1) receptor signalling. PMID- 17920584 TI - Preclinical pharmacology profile of CS-706, a novel cyclooxygenase-2 selective inhibitor, with potent antinociceptive and anti-inflammatory effects. AB - We report here the preclinical anti-inflammatory profile of CS-706 [2-(4 ethoxyphenyl)-4-methyl-1-(4-sulfamoylphenyl)-1H-pyrrole], a novel cyclooxygenase 2 (COX-2) selective inhibitor. CS-706 selectively inhibited COX-2 in a human whole blood assay with an IC(50) of 0.31 microM, compared with an IC(50) of 2.2 microM for COX-1. The selectivity ratio of CS-706 was higher than those of the conventional non-steroidal anti-inflammatory drugs naproxen, indomethacin, and Diclofenac-Na, whereas it was lower than those of rofecoxib, valdecoxib and etoricoxib. It was similar to that of celecoxib. The pharmacokinetic profile of CS-706 showed rapid absorption and dose-proportional exposure after oral administration to rats. CS-706 inhibited prostaglandin E(2) production in inflamed tissue induced by yeast-injection in rats with potency similar to that of indomethacin. However, it inhibited gastric mucosal prostaglandin E(2) production in normal rats weakly compared with indomethacin. CS-706 ameliorated both yeast-induced inflammatory acute pain (ED(50)=0.0090 mg/kg) and adjuvant induced chronic arthritic pain (ED(50)=0.30 mg/kg) in rats. CS-706 showed more potent antinociceptive activity than celecoxib and rofecoxib in these models. In an adjuvant-induced arthritic model in rats, CS-706 suppressed foot swelling prophylactically with an ID(50) of 0.10 mg/kg/day, and decreased foot swelling in the established arthritis therapeutically in a dose range of 0.040 to 1.0 mg/kg/day. Single administration of up to 100 mg/kg of CS-706 induced no significant gastric lesions in rats. In conclusion, CS-706 is a COX-2-selective inhibitor with a potent antinociceptive and anti-inflammatory activity and a gastric safety profile. PMID- 17920586 TI - Deficiency in monocyte chemoattractant protein-1 modifies lipid and glucose metabolism. AB - We describe the effect of MCP-1 deficiency in mice rendered hyperlipemic by the concomitant ablation of the LDL receptor. The MCP-1(-/-)LDLr(-/-) mice in comparison with LDLr(-/-) mice showed a decreased lipoprotein clearance, derangements in free fatty acids delivery and less glucose tolerance when fed a regular chow, and they showed a partial resistance to alterations in glucose and lipid metabolism induced by dietary fat and cholesterol. They also were less prone to the development of diet-induced obesity. Our results suggest that the role of MCP-1 in metabolism is relevant and that, although new hidden complexities are evident, the function of MCP-1/CCL2 extends far beyond the monocyte chemoattractant effect. Therefore, the regulatory mechanisms influenced by MCP-1 should be fully ascertained to understand the metabolic consequences of inflammation and before considering MCP-1 as a therapeutic target. PMID- 17920585 TI - Melatonin reduces formalin-induced nociception and tactile allodynia in diabetic rats. AB - The purpose of this study was to assess the antinociceptive and antiallodynic effect of melatonin as well as its possible mechanism of action in diabetic rats. Streptozotocin (50 mg/kg) injection caused hyperglycemia within 1 week. Formalin evoked flinching was increased in diabetic rats as compared to non-diabetic rats. Oral administration of melatonin (10-300 mg/kg) dose-dependently reduced flinching behavior in diabetic rats. In addition, K-185 (a melatonin MT(2) receptor antagonist, 0.2-2 mg/kg, s.c.) completely blocked the melatonin-induced antinociception in diabetic rats, whereas that naltrexone (a non-selective opioid receptor antagonist, 1 mg/kg, s.c.) and naltrindole (a selective delta opioid receptor antagonist, 0.5 mg/kg, s.c.), but not 5'-guanidinonaltrindole (a selective kappa opioid receptor antagonist, 1 mg/kg, s.c.), partially reduced the antinociceptive effect of melatonin. Given alone K-185, naltrexone, naltrindole or 5'-guanidinonaltrindole did not modify formalin-induced nociception in diabetic rats. Four to 8 weeks after diabetes induction, tactile allodynia was observed in the streptozotocin-injected rats. On this condition, oral administration of melatonin (75-300 mg/kg) dose-dependently reduced tactile allodynia in diabetic rats. Both antinociceptive and antiallodynic effects were not related to motor changes as melatonin did not modify number of falls in the rotarod test. Results indicate that melatonin is able to reduce formalin-induced nociception and tactile allodynia in streptozotocin-injected rats. In addition, data suggest that melatonin MT(2) and delta opioid receptors may play an important role in these effects. PMID- 17920587 TI - FERM protein EPB41L5 is a novel member of the mammalian CRB-MPP5 polarity complex. AB - Cell polarity is induced and maintained by separation of the apical and basolateral domains through specialized cell-cell junctions. The Crumbs protein and its binding partners are involved in formation and stabilization of adherens junctions. In this study, we describe a novel component of the mammalian Crumbs complex, the FERM domain protein EPB41L5, which associates with the intracellular domains of all three Crumbs homologs through its FERM domain. Surprisingly, the same FERM domain is involved in binding to the HOOK domain of MPP5/PALS1, a previously identified interactor of Crumbs. Co-expression and co-localization studies suggested that in several epithelial derived tissues Epb4.1l5 interacts with at least one Crumbs homolog, and with Mpp5. Although at early embryonic stages Epb4.1l5 is found at the basolateral membrane compartment, in adult tissues it co-localizes at the apical domain with Crumbs proteins and Mpp5. Overexpression of Epb4.1l5 in polarized MDCK cells affects tightness of cell junctions and results in disorganization of the tight junction markers ZO-1 and PATJ. Our results emphasize the importance of a conserved Crumbs-MPP5-EPB41L5 polarity complex in mammals. PMID- 17920588 TI - Systematic evaluation of ICG and trypan blue related effects on ARPE-19 cells in vitro. AB - The aim of our study was a systematic analysis of the impact of variable parameters on indocyanine green (ICG) and trypan blue (TB) related cytotoxicity on human RPE cells. ARPE-19 cells were incubated with ICG (5.0-0.025mg/ml), with ICG-free solutions of corresponding osmolarities or with TB (1.5-0.0375mg/ml). Incubation lasted 1-20min with or without endolight illumination for 1min or 5min. Cell viability and morphology were examined after 6h, 24h and 72h to detect acute and delayed effects. In the absence of endolight, ICG cytotoxicity depends on osmolarity and exposure time. In the presence of endolight, cytotoxic effects are influenced by dye concentration. TB cytotoxicity depends on dye concentration and exposure time, but not on illumination. All observed cytotoxic effects were mainly acute. Both ICG and TB can be cytotoxic depending on concentration and exposure time. ICG related cytotoxic effects are additionally determined by osmolarity and phototoxicity. However, concentrations (<1mg/ml) and incubation times (<5min) as used in clinical practice would appear to be well tolerated. PMID- 17920589 TI - Effects of thalamic stimulation frequency on intention and postural tremor. AB - Deep brain stimulation (DBS) of the ventral intermediate nucleus (VIM) of the thalamus improves essential tremor. Suppression of the amplitude of the postural tremor component with VIM DBS depends on stimulation frequency. The purpose of this study was to determine the effect of DBS frequency on the intention tremor component, that is, tremor that is enhanced by target-directed movement, and to compare it to the effect of DBS frequency on postural tremor in people with essential tremor. We measured tremor frequency and amplitude during trials of postural holding and voluntary reaching between two targets at 10 different stimulation frequency settings between 0 and 185 Hz. Tremor frequency did not change with changes in stimulation frequency. Amplitude suppression of both intention and postural tremor depended on stimulation frequency. Maximal tremor reduction occurred at approximately 130 Hz for both forms of tremor. However, at optimal frequencies, the percent reduction in tremor amplitude relative to the DBS OFF condition was greater for postural than for intention tremor. These results suggest that VIM DBS stimulation frequencies near 130 Hz may provide maximal control of intention and postural tremor. Identification of optimal stimulation settings should consider assessment of intention tremor, not just postural tremor, as intention tremor may not be as well controlled as postural tremor but may be a better gauge for functional benefit. PMID- 17920590 TI - Design of clinical trials of gene therapy in Parkinson disease. AB - No current therapy for Parkinson disease has been shown to slow or reverse the progressive course of the disease. As a departure from traditional treatments, gene therapy approaches provide a new hope for realizing this long-sought goal; but before they can be widely employed for use in patients, they must first be submitted to the rigorous safety and efficacy standards of the clinical trial. Some of the challenges of gene therapy clinical trial design are similar to those in studies of conventional pharmacological agents and include addressing the heterogeneity of the disease, the need for clinical and surrogate endpoints, and the issue of distinguishing "symptomatic" from "neuroprotective" effects. Gene therapy trials also raise the issues of the risks of viral therapy, issues of dose-response, the need for sham surgery, and the long duration of risks and benefits. We conclude that the most feasible designs are for those treatments that are expected to produce a rapid improvement in directly observable symptoms. Trials of agents which are expected to produce only a slowing of progression and not a reversal of the disease course are likely to take much longer and will require the development of methods to assess quality of life and other non-motor aspects of the disease. PMID- 17920591 TI - Plasmodium falciparum: functional mitochondrial ADP/ATP transporter in Escherichia coli plasmic membrane as a tool for selective drug screening. AB - Plasmodium falciparum mitochondrial ADP/ATP transporter or adenylate translocase (PfAdT) was previously characterised at the molecular level and intracellularly located by immuno-electromicroscopy. Inhibition of this transporter blocks parasite development in erythrocytes. In this study, PfAdT was expressed in C43 (DE3) Escherichia coli strain under isopropyl beta-d-thiogalacto-pyranoside (IPTG) induction to screen inhibitory molecules. PfAdT was integrated directly into the bacterial cytoplasmic membrane. Whereas IPTG-induced bacterial cells imported radioactively labelled ATP, non-induced cells did not. The transporter bound specifically ADP and ATP, but not AMP. IPTG-induced cells preloaded with labelled ATP exported ATP after exogenous addition of unlabelled ADP or ATP, indicating a counter exchange transport mechanism. Bongrekic acid and atractyloside, two well-known specific inhibitors of mitochondrial ADP/ATP transporter, were tested. This experimental model was evaluated using three Malagasy crude plants extracts which have shown antiplasmodial activity on in vitro parasite cultures. PMID- 17920592 TI - Spotlight on... Barry Halliwell. Interview by Daniela Ruffell. PMID- 17920593 TI - A case of hydrosalpinx associated with the menstrual cycle. AB - OBJECTIVE: To describe a case report of hydrosalpinx that changed dramatically in size during the menstrual cycle. DESIGN: Case report. SETTING: University teaching hospital reproductive endocrinology and infertility practice. PATIENT(S): A 32-year-old woman with a history of medical and surgical treatments of endometriosis who sought infertility treatment. INTERVENTION(S): Transvaginal ultrasonography performed sequentially during menstrual cycles. MAIN OUTCOME MEASURE(S): Size of hydrosalpinx-like image. RESULT(S): The size of the hydrosalpinx-like image in the left adnexal region varied; it peaked during the ovulatory period and then remarkably diminished in a cyclic manner. Laparoscopy revealed a dense adhesion between the left tubal fimbriated end and the posterior uterine wall, which led to terminal obstruction of the tube. CONCLUSION(S): Change in the volume of the hydrosalpinx in this case was speculated to reflect the normal tubal fluid production regulated by ovarian hormones. PMID- 17920594 TI - Surgical correction of rare Mullerian anomalies and spontaneous pregnancies: a case report. AB - OBJECTIVE: To surgically reconstruct rare Mullerian duct anomalies with varying degrees of bladder exstrophy and congenital absence of ampullary portion of fallopian tube. DESIGN: Case report. SETTING: Institute of Reproductive Medicine, Salt Lake, Kolkata, India. PATIENT(S): Two cases of anteriorly displaced vagina with stenotic introitus, divergent pubic rami, and varying degrees of bladder exstrophy (group A) and three cases of congenital absence of ampullary portion of the fallopian tube with intact but blind fimbria attached to mesoovarium (group B). INTERVENTION(S): Introitoplasty and tubal anastomosis were performed for groups A and B, respectively. MAIN OUTCOME MEASURE(S): Description and treatment of two types of patients with rare Mullerian anomalies. RESULT(S): Three spontaneous pregnancies and viable deliveries, two in group A and one in group B, were achieved after surgical reconstruction. CONCLUSION(S): Assisted reproductive technologies usually provide a solution to infertility in rare Mullerian anomalies; reconstructive surgical procedures specific to the type of anatomical defect may also be considered for spontaneous pregnancy with viable deliveries. PMID- 17920595 TI - Cryopreservation of oocytes in a young woman with severe and symptomatic endometriosis: a new indication for fertility preservation. AB - OBJECTIVE: To report a new indication for fertility preservation. DESIGN: Case report. SETTING: Academic teaching hospital. PATIENT(S): A 25-year-old nulliparous woman with severe and symptomatic endometriosis and low antral follicular count. INTERVENTION(S): Oocyte cryopreservation. MAIN OUTCOME MEASURE(S): Number of cryopreserved oocytes. RESULT(S): After three cycles of ovarian stimulation, we cryopreserved 21 oocytes. CONCLUSION(S): We recommend fertility preservation as part of preoperative counseling in young women with severe endometriosis. PMID- 17920596 TI - Effects of ovine CRF injections into the dorsomedial, dorsolateral and lateral columns of the periaqueductal gray: a functional role for the dorsomedial column. AB - Corticotropin-releasing factor (CRF) and its receptor subtypes have been implicated in the regulation of endocrine, behavioral and autonomic responses to stress, fear and anxiety. Ovine CRF (oCRF) is a nonspecific CRF receptor agonist that produces anxiogenic-like effects when injected locally into the dorsal aspects of the periaqueductal gray (PAG). This structure is subdivided into four distinct longitudinal columns but their exact functional role is not fully understood. The purpose of the present study was to characterize the effects of oCRF (0.25, 0.5 and 1 microg/0.2 microL) injections into the dorsomedial (dmPAG), dorsolateral (dlPAG) and lateral (lPAG) columns of the PAG using an analysis of the exploratory behavior of rats in the elevated plus-maze (EPM) test. The results showed that microinjections of oCRF intra-dmPAG reduced entries and time spent in the open arms and decreased end-arm exploration and head-dipping. In contrast, oCRF intra-dlPAG or lPAG did not affect the exploratory behavior of the animals in the EPM. These findings point to a columnar specificity for the oCRF effects in the PAG, that is, it increased spatial avoidance measures of the EPM test only in the dmPAG. The proaversive effects of oCRF in the dmPAG gain further relevance when combined with previous immunohistochemical studies showing that CRF-containing projections from the periventricular hypothalamic system arch dorsomedially to the PAG, which could function as an important relay station in the midbrain tectum for avoidance behaviors. PMID- 17920597 TI - Spontaneous egg or chick abandonment in energy-depleted king penguins: a role for corticosterone and prolactin? AB - Various exogenous or endogenous factors may induce an emergency response in birds, redirecting current activity towards survival. In fasting, breeding penguins, the achievement of a critical energy depletion was suggested to induce egg abandonment and departure to sea for re-feeding. How such a behavioral shift is hormonally controlled remains unknown. The possible involvement of corticosterone and prolactin was examined by characterizing the nutritional and hormonal states of king penguins at egg abandonment. Further, we tested if these states differ according to whether an egg or a chick is abandoned, and according to the timing of breeding. In every case of abandonment, birds were in phase III fasting characterized by accelerated protein catabolism. However, body condition at egg abandonment was lower in early than in late breeders, suggesting that king penguins are willing to tolerate a larger energy depletion when their potential breeding success is high. At egg and chick abandonment, plasma corticosterone levels were, respectively, increased by 2- and 4-fold, whereas plasma prolactin levels were, respectively, depressed by 3- and 1.4-fold. The increase in plasma corticosterone and the decrease in plasma prolactin could be involved in the control of abandonment by, respectively, stimulating the drive to re-feed and diminishing the drive to incubate or brood. The smaller decrease in prolactin levels and the greater increase in corticosterone levels observed at chick vs egg abandonment suggest that, in addition to nutritionally-related stimuli, tactile or audible stimuli from the egg or chick could intervene in the endocrine control of abandonment. PMID- 17920598 TI - Auditory stimulation of reproductive function in male Rufous-winged Sparrows, Aimophila carpalis. AB - Prolonged exposure to conspecific song stimulates gonadal function and reproductive hormone secretion in female birds but few studies have investigated the physiological effects of conspecific song exposure on males outside of short term, aggressive interactions. We exposed male Rufous-winged Sparrows, Aimophila carpalis, either to conspecific song (CS Song), to heterospecific song (Black throated Sparrow, Amphispiza bilineata; HS Song), or to no recorded song (No Song) for 59 consecutive days (two h per day). Birds were exposed to short days (8L:16D) for the first 21 days of treatment and were then transferred to long days (13L:11D) for the remaining 38 days. During long day exposure, CS Song birds experienced faster growth of testes than HS Song and No Song birds. HS Song birds also grew their testes faster than No Song birds. Plasma luteinizing hormone (LH) and testosterone did not differ between CS Song and No Song birds. However, plasma LH was higher in HS Song birds compared to other groups. There were no differences in hypothalamic immunocytochemical labeling for gonadotropin releasing hormone, its precursor proGnRH, or gonadotropin-inhibitory hormone, nor were there differences in two song control nuclei volumes (HVC and RA) between CS Song and No Song treatment groups. Furthermore, we found no effect of heterospecific song on free-living Rufous-winged Sparrow aggressive behaviors. These data indicate that long-term exposure to auditory stimuli, such as song, can influence the reproductive system of male songbirds and different types of auditory stimuli can have differential effects on reproductive function. PMID- 17920599 TI - A comparison of liquid-based cytology with conventional cytology. AB - OBJECTIVE: To evaluate the 2 methods of cytologic screening to detect abnormalities of the cervical epithelium. METHODS: This study with 3 groups of women was performed at Selcuk University Meram Medical School between January 2004 and March 2006. In one group (paired sample for specimen collection) women were screened with conventional cytology; in another group (paired sample for specimen collection) they were screened with liquid-based cytology; and in the third group (split sample for specimen collection) they were screened by both methods. RESULTS: The rate of unsatisfactory results was lower in the liquid based than in the conventional cytology group (6.1% vs. 2.6%; P<0.05). More smears containing atypical squamous cells of undetermined significance were detected by the liquid-based method, but the difference was not statistically significant. Also, no statistically significant differences between liquid-based and conventional cytology were observed in the detection of other epithelial abnormalities (P>0.05). CONCLUSION: The liquid-based and conventional cytology methods were found to be equivalent in the detection of cervical epithelial abnormalities. PMID- 17920600 TI - Maternal mortality surveillance in Jamaica. AB - OBJECTIVES: To assess factors associated with under-reporting of maternal deaths from 1998, when maternal deaths became a Class I notifiable event in Jamaica and continuous maternal mortality surveillance was introduced, through 2003. METHODS: The number of deaths notified was compared with the number of independently identified deaths for each period and region studied, and key informants reported on their experience of the surveillance process. RESULTS: By 2000, approximately 80% of maternal deaths were reported, and was more consistent in 2 of the 4 regions. In these 2 regions someone was responsible for active surveillance and there was an established maternal mortality committee to review cases. Factors associated with nonreporting were no postmortem examination, death in the first trimester of pregnancy, and time interval between pregnancy termination and death. The surveillance staff requested guidelines on monitoring interregional transfers and technical assistance in developing action plans. CONCLUSION: Active hospital surveillance must include all wards, including the emergency department. Community surveillance should include forensic pathologists. National leadership is needed to summarize trends, address policy, and provide technical assistance to the surveillance staff. PMID- 17920602 TI - Definitions of mediolateral episiotomy in Europe. PMID- 17920601 TI - Antenatal detection of arteriovenous anastomoses in monochorionic twin pregnancy. AB - OBJECTIVES: To revalidate the detection technique for arteriovenous anastomoses in an unselected group of monochorionic twins, and to make recommendations about its applicability for more widespread use. METHODS: Women with monochorionic diamniotic (MCDA) twins were recruited and underwent placental mapping by color Doppler ultrasound. Images of placental maps showing the location and type of anastomoses were saved as digital video clips. After delivery, dye injection study of all the placentas was performed to delineate the site and type of anastomoses. A digital photograph of each injection study was taken and saved. The antenatal ultrasound images and postnatal dye injection studies were compared. RESULTS: 18 sets MCDA twins were evaluated. In 3 cases there was evidence of twin-to-twin transfusion syndrome. Dye injection of 18 placentas revealed 21 arterio-arterial anastomoses (AAA), 21 arteriovenous anastomoses (AVA) and 4 veno-venous anastomoses (VVA). Of these, 10 (48%) AAAs and 5 (24%) AVAs were detected antenatally by color Doppler. In all cases, where an AVA was detected, the placenta was located anteriorly. CONCLUSION: Antenatal detection of AVA was feasible when the placenta was located anteriorly, but proved difficult in posteriorly situated placentas. PMID- 17920603 TI - Uterine rupture presenting as a vaginal prolapse of gangrenous small bowel. PMID- 17920604 TI - Digital and transvaginal ultrasound cervical assessment for prediction of successful labor induction. AB - OBJECTIVE: To compare the values of preinduction transvaginal cervical length measurements and Bishop score to predict successful labor induction. METHODS: A prospective, observational trial of nulliparous women undergoing labor induction. Inclusion criteria were gestational age between 36 and 42 weeks, singleton cephalic presentation of the fetus, and intact membranes. Preinduction cervical ripening was performed using 25 microg intravaginal misoprostol (PGE1), repeated every 4 h, up to a maximum of 3 doses. Induction was subsequently continued by oxytocin and amniotomy. RESULTS: A total of 43 women met the inclusion criteria. Mean preinduction cervical length for women with successful or failed labor induction was 26 mm (95% Confidence interval [CI], 27-32) and 34 mm (95% CI, 33 38), respectively (P=0.002). Mean Bishop scores for successful and failed induction groups were 5.4 (95% CI, 5.2-6.2) and 3.1 (95% CI, 2.8-3.5), respectively (P=0.003). CONCLUSION: Digital examination and transvaginal ultrasound of the cervix predict successful labor induction with reasonable accuracy. PMID- 17920606 TI - Two-dimensional hydrophilic interaction chromatography coupling anion-exchange and hydrophilic interaction columns for separation of 2-pyridylamino derivatives of neutral and sialylated N-glycans. AB - A novel chromatographic approach coupling anion-exchange (diethylaminoethylene) and hydrophilic-interaction (amide or zwitterionic type) columns was developed for the separating of 2-pyridylamino derivatives of N-glycans (PA-N-glycans). This is a kind of on-line, two-dimensional (2D) separation approach in hydrophilic-interaction chromatography (called the 2D-HILIC method), analogous to that of coupling cation- (or anion-, or mixed ion-) exchange and reversed-phase columns in hydrophobic interaction (reversed-phase) chromatography. The efficiency of the 2D-HILIC method was tested with biantennary neutral and sialylated PA-N-glycan standards by properly combining linear gradient elutions of water-acetonitrile and spiked-salt (ammonium acetate) elutions. The retention time RSDs of all the peaks in three sequential runs of a 100 min cycle are less than 0.52%, which indicates a reasonably good repeatability of the 2D-HILIC method. Then, the method was applied to a complex mixture of PA-N-glycans from human serum proteins. It was demonstrated that the neutral PA-N-glycans and mono , di-, tri-, and tetrasialylated PA-N-glycans are able to be eluted in turn according to the number of sialic acids in an automated (programmed) single run. PMID- 17920605 TI - Bayesian spatial analysis of a national urinary schistosomiasis questionnaire to assist geographic targeting of schistosomiasis control in Tanzania, East Africa. AB - Spatial modelling was applied to self-reported schistosomiasis data from over 2.5 million school students from 12,399 schools in all regions of mainland Tanzania. The aims were to derive statistically robust prevalence estimates in small geographical units (wards), to identify spatial clusters of high and low prevalence and to quantify uncertainty surrounding prevalence estimates. The objective was to permit informed decision-making for targeting of resources by the Tanzanian national schistosomiasis control programme. Bayesian logistic regression models were constructed to investigate the risk of schistosomiasis in each ward, based on the prevalence of self-reported schistosomiasis and blood in urine. Models contained covariates representing climatic and demographic effects and random effects for spatial clustering. Degree of urbanisation, median elevation of the ward and median normalised difference vegetation index (NDVI) were significantly and negatively associated with schistosomiasis prevalence. Most regions contained wards that had >95% certainty of schistosomiasis prevalence being >10%, the selected threshold for bi-annual mass chemotherapy of school-age children. Wards with >95% certainty of schistosomiasis prevalence being >30%, the selected threshold for annual mass chemotherapy of school-age children, were clustered in north-western, south-western and south-eastern regions. Large sample sizes in most wards meant raw prevalence estimates were robust. However, when uncertainties were investigated, intervention status was equivocal in 6.7-13.0% of wards depending on the criterion used. The resulting maps are being used to plan the distribution of praziquantel to participating districts; they will be applied to prioritising control in those wards where prevalence was unequivocally above thresholds for intervention and might direct decision-makers to obtain more information in wards where intervention status was uncertain. PMID- 17920607 TI - Protein aggregation kinetics during Protein A chromatography. Case study for an Fc fusion protein. AB - Protein A chromatography has come to be widely adopted for large-scale purification of monoclonal antibodies and Fc fusion proteins. The low pH conditions required for Protein A elution can often lead to aggregation issues for these products. A concerted study of the kinetics of aggregate formation and their relation to chromatography on Protein A media has been lacking. This paper provides a framework to describe aggregation kinetics for an Fc fusion protein that was highly susceptible to aggregate formation under low pH conditions. In contrast to what is usually expected to be a higher order reaction, first order aggregation kinetics were observed for this protein over a wide range of conditions. A comparison of the rate constants of aggregation forms an effective means of comparing various stabilizing additives to the elution buffer with one another. Inclusion of urea in the elution buffer at moderate concentrations (<2M) and low temperature operation of the Protein A column were both found to be effective solutions to the aggregation issue. Elution from the Protein A resin was found to increase the aggregation rate constants over and above what would be expected from exposure to low pH conditions in solution alone. This demonstrates that Protein A-Fc interactions can destabilize product structure and increase the tendency to aggregate. The results presented here are anticipated to assist the development of Protein A process conditions for products that are prone to form high molecular weight aggregates during column elution. PMID- 17920608 TI - Semiochemicals of the Scarabaeinae VIII. Identification of active constituents of the abdominal sex-attracting secretion of the male dung beetle, Kheper bonellii, using gas chromatography with flame ionization and electroantennographic detection in parallel. AB - Using gas chromatography with flame ionization detection and electroantennographic detection in parallel (GC-FID/EAD), the active constituents of the sex attractant of male dung beetles of Kheper bonellii were located in the gas chromatogram of an extract of the secretion. These constituents were identified as propanoic acid, butanoic acid, indole, 3-methylindole (skatole) and methyl cis-cascarillate (methyl cis-2-2'-hexylcyclopropylacetate) by, inter alia, GC-MS, (1)H and (13)C NMR analysis, and synthesis. These compounds elicited EAD responses in male as well as female antennae. Racemic methyl cis-cascarillate was synthesized for comparison with the natural methyl ester. Enantioselective GC FID/EAD using a capillary column coated with OV-1701-OH containing 10% heptakis(2,3-di-O-methyl-6-O-tert-butyldimethylsilyl)-beta-cyclodextrin showed that the natural compound co-eluted with the first-eluting enantiomer of the racemic methyl cis-cascarillate, which was the only enantiomer that elicited EAD responses in the antennae of male and female K. bonellii. The absolute configuration of this enantiomer was established by a stereoselective synthesis, which gave methyl (R,R)-cascarillate [methyl (1'R,2'R)-2-2' hexylcyclopropylacetate] in an enantiomeric excess of 69%. PMID- 17920609 TI - Improved cleanup technique for gas chromatographic-mass spectrometric determination of alkylphenols from biota extract. AB - A simple and economical cleanup technique was developed to determine alkylphenols by GC-MS from biological extracts containing relatively high lipids. The lipids were successfully removed from bivalve extracts through a two-step cleanup. The new method is a combination of Florisil adsorption chromatography and silyl derivatization technique. Low and high (non-polar and highly polar) molecular weight lipids were removed from the biota extract with deactivated Florisil column in the first step. And in the second step, middle molecular weight (middle polar) lipids were removed in an activated Florisil column after the alkylphenols were converted to corresponding silyl derivatives with bis(trimethylsilyl)trifluoroacetamide (BSTFA). On the basis of the above results, a simple cleanup kit was developed for convenience. The technique was optimized with reference to the activity of packing materials and polarity of eluting solvents. Only 3g of Florisil, 25 mL of hexane and 10 mL of dichloromethane were required for one sample. The recoveries of alkylphenols from spiked samples varied from 88 to 103% with a low relative standard deviation (mean value: 5.3%) and the recovery was similar or even higher than other methods currently in use. The technique was successfully applied to mussel samples from Masan Bay, South Korea. Simultaneous measurement of these compounds in water, sediment and biota; the resulting bio-concentration factor and their relationships confirm previously published works, validating the method applied. PMID- 17920610 TI - Determination of functionalised carboxylic acids in atmospheric particles and cloud water using capillary electrophoresis/mass spectrometry. AB - A capillary electrophoresis/electrospray ionisation mass spectrometry (CE/ESI-MS) method was developed for the determination of 38 organic acids in atmospheric particles and cloud water. The target analytes include many functionalised carboxylic acids, such as carboxylic acids with additional oxo-, hydroxy- or nitro-groups. These compounds are of large interest as their determination might give new insights into the atmospheric multiphase chemistry. OASIS HLB sorbent material (Waters) was used to extract and enrich polar carboxylic acids from aqueous solutions with recoveries greater than 80% for most analytes. Relative standard deviations in the range of 4-20% for peak areas (n=5), including the SPE step, and 0.2-0.5% (n=8) for migration times were found. The limits of detection (S/N=3) ranged from 0.005 to 0.6 micromol l(-1) for an ion-trap mass spectrometer and from 0.0004 to 0.08 micromol l(-1) for a time-of-flight mass spectrometer. These detection limits translate into atmospheric concentrations in the low pg m( 3) range based on the experimental conditions in this study. Severe matrix effects were observed for real samples, arising from complex co-extracted organic material. However, using the method of standard addition, most of the analytes could successfully be quantified in samples of ambient particles and cloud water with concentrations in the low ng m(-3) to high pg m(-3) range. These results demonstrate the suitability of the proposed method for the determination of a wide range of polar carboxylic acids at low concentrations in complex samples of different atmospheric phases. PMID- 17920611 TI - Determination of selected antioxidants in Melissae herba by isotachophoresis and capillary zone electrophoresis in the column-coupling configuration. AB - A method utilising isotachophoresis and capillary zone electrophoresis in the column coupling configuration with UV detection at 320 nm was developed for separation and determination of five phenolic acids (rosmarinic, p-coumaric, ferulic, caffeic and chlorogenic) and flavonoid quercitrin in a methanolic extract of Melissae herba. The proposed method has been validated with correlation coefficients from 0.9842 to 0.9988, RSD values between 0.39% and 0.83% for migration times and between 0.40% and 2.05% for peak areas. PMID- 17920612 TI - Use of in-tube sorptive extraction techniques for determination of benzene, toluene, ethylbenzene and xylenes in soft drinks. AB - A comparison is made between static headspace analysis and headspace solid-phase dynamic extraction (HS-SPDE) for the quantitative determination of trace level BTEX solvents (benzene, toluene, ethylbenzene and o-, m-, and p-xylene) in soft drinks. Two non-polar extraction phases were investigated for SPDE using an automated sampler with a gas-tight syringe equipped with a special needle coated on the inside with the extraction phase. Following adsorption onto the phase, the analytes were thermally desorbed directly into a GC-MS. The techniques were optimised and evaluated by analysis of spiked soft drink samples. The use of the SPDE device gave comparable results to the static headspace method, with lower detection limits for some compounds, and also offers advantages for applications where lower temperatures are preferred. PMID- 17920613 TI - Fast gas chromatography for pesticide residues analysis using analyte protectants. AB - Fast GC-MS with narrow-bore columns combined with effective sample preparation technique (QuEChERS method) was used for evaluation of various calibration approaches in pesticide residues analysis. In order to compare the performance of analyte protectants (APs) with matrix-matched standards calibration curves of selected pesticides were searched in terms of linearity of responses, repeatability of measurements and reached limit of quantifications utilizing the following calibration standards in the concentration range 1-500 ng mL(-1)(the equivalent sample concentration 1-500 microg kg(-1)): in neat solvent (acetonitrile) with/without addition of APs, matrix-matched standards with/without addition of APs. For APs results are in a good agreement with matrix matched standards. To evaluate errors of determination of concentration synthetic samples at concentration level of pesticides 50 ng mL(-1) (50 microg kg(-1)) were analyzed and quantified using the above given standards. For less troublesome pesticides very good estimation of concentration was obtained utilizing APs, while for more troublesome pesticides such as methidathion, malathion, phosalone and deltamethrin significant overestimation reaching up to 80% occurred. According to presented results APs can be advantegously used for "easy" pesticides determination. For "difficult" pesticides an alternative calibration approach is required for samples potentially violating MRLs. An example of real sample measurement is shown. In this paper also the use of internal standards (triphenylphosphate (TPP) and heptachlor (HEPT)) for peak areas normalization is discussed in terms of repeatability of measurements and quantitative data obtained. TPP normalization provided slightly better results than the use of absolute peak areas measurements on the contrary to HEPT. PMID- 17920614 TI - Determination of saccharides in atmospheric aerosol using anion-exchange high performance liquid chromatography and pulsed-amperometric detection. AB - An improved method is described for the quantification of primary sugars, sugar alcohols and anhydrosugars in atmospheric aerosols, making use of separation by high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD). Quartz fibre filters from high-volume samplers were extracted with water and the extract injected directly. Repeatability is typically 4% RSD, for e.g. levoglucosan at 50 ng m(-3) in air, better for winter levels around 700 ng m(-3). Limits of detection for individual sugars are in the range 0.02-0.05 microg mL(-1) in solution, corresponding to 2-5 ng m(-3) from a 20 m(3) air sample. The overlap of arabitol and levogluocosan is overcome by using a Dionex PA-1 column, with appropriate control of eluent composition, and peak deconvolution software, allowing quantification of both sugars in difficult summer samples containing low-levels of levoglucosan. Analysis of a set of ambient aerosol samples by both GC-flame ionization detection and HPAEC-PAD shows good agreement. The new method has the advantage of requiring no sample pretreatment or derivatization and is thus well suited to handling large numbers of samples. PMID- 17920615 TI - Developments in the use and fabrication of organic monolithic phases for use with high-performance liquid chromatography and capillary electrochromatography. AB - Capillary electrochromatography suffered in its development because of difficulty in producing stable columns with good permeability. Variability in frit characteristics gave rise to non-reproducible capillaries whose fabrication was extremely difficult and time consuming. Monolithic stationary phases gained popularity in the early 1990s due to the fact that they were easy to fabricate and required no retaining frits. They were also able to be manufactured in a wide variety of chemistries which made them very interesting to the analytical chemist who is constantly looking for materials with different selectivity to the popular silica-based stationary phases. PMID- 17920616 TI - Synthesis of composite particles through emulsion polymerization based on silica/fluoroacrylate-siloxane using anionic reactive and nonionic surfactants. AB - The composite particles with core/shell structure resulting from the combination of silica seed and hydrophobic copolymer (dodecafluoroheptyl methacrylate (DFMA), gamma-methacryloxypropyltriisopropoxidesilane (MAPTIPS), methyl methacrylate, butyl acrylate) were synthesized by emulsion polymerization. The amount of the silica seeds, concentration of reactive surfactant, as well as the addition of DFMA and MAPTIPS, have strong influences on the morphology of composite particles. It has been shown that it would be possible to produce stable organic/inorganic composite particles with inhomogeneous core/shell structure encapsulated by hydrophobic fluorinated acrylate even though using unmodified silica particles and admixture of anionic and nonionic surfactants. However, there was an obvious difference on the morphologies of core-shell structure whether the DFMA and MAPTIPS were added or not. It was concluded that two kinds of polymerization approaches might coexist in the presence of DFMA and MAPTIPS for raw silica. One clear advantage of this process is that there is only one silica bead for each composite particle. This kind of stable core-shell structural hybrid latex is useful for preparing high performance hydrophobic coating. PMID- 17920617 TI - How smart do you need to be to get it wrong? The role of cognitive capacity in the development of heuristic-based judgment. AB - We examined the relationship between cognitive capacity and heuristic responding on four types of reasoning and decision-making tasks. A total of 84 children, between 5 years 2 months and 11 years 7 months of age, participated in the study. There was a marked increase in heuristic responding with age that was related to increases in cognitive capacity. These findings are inconsistent with the predominant dual-process accounts of reasoning and decision making as applied to development. We offer an alternative explanation of the findings, considering them in the context of recent claims concerning the role of working memory in contextualized reasoning. PMID- 17920618 TI - There's something about obesity: culture, contagion, rationality, and children's responses to drinks "created" by obese children. AB - Theories of the development of obesity stereotypes cannot easily explain the stigma associated with being obese. Evidence that important similarities exist between the symptoms of obesity and contagious illnesses, young children have "theories" of illnesses, and obesity stereotypes are among the earliest that children develop led to the hypothesis that children would find beverages purportedly created by obese children less tasteful and more memorable than beverages created by average weight children. After assignment to two story conditions in which a child became ill after eating an unfamiliar food, Caucasian American and Chinese 7- and 10-year-olds sampled identically flavored "obese created" and "average-created" beverages. Taste ratings were lower, ratings of the chances of feeling sick were higher, and memory was superior for obese created drinks than for average-created drinks, particularly when the character in the story contracted a contagious illness and memory was scored for "gist." Finally, children often created the false memory that the story character was an obese beverage creator. The roles of contagion and magical beliefs are discussed, as are the rationality of children's responses and the relevance of the findings for theories of obesity stereotypes. PMID- 17920619 TI - Simultaneous analysis of multiple serum proteins adhering to the surface of medical grade polydimethylsiloxane elastomers. AB - Although polydimethylsiloxane (PDMS, silicone) elastomers are presumed to be chemically inert and of negligible toxicity, they induce a prompt acute inflammatory response with subsequent fibrotic reactions. Since local inflammatory and fibrotic side effects are associated with the proteinaceous film on the surface of silicone implants, the process of protein adherence to silicone is of practical medical relevance, and interesting from theoretical, clinical and biotechnological perspectives. It is hypothesized that the systemic side effects resembling rheumatoid and other connective tissue diseases may be triggered by local immunological changes, but this functional relationship has yet to be defined. Because the proteinaceous film on the surface of silicone has been identified as a key player in the activation of host defense mechanisms, we propose a test system based on a proteomics screen to simultaneously identify proteins adsorbed from serum to the surface of silicone. Herein, we describe protein adsorption kinetics on the surface of silicone implants, correlate the adhesion properties of serum proteins with the occurrence of adverse reactions to silicone, and successfully discriminate their signature on the silicone surface in a blinded study of patients suffering from fibrotic reactions (as determined by Baker scale) to silicone implants. PMID- 17920620 TI - A new simple multi-well plate-based assay for monocyte differentiation using human monocytic leukemia THP-1 cells. AB - A new simple multi-well plate-based assay to determine monocyte differentiation inducing activity was developed. It is based on the increase of macrophage adherence after the induction of monocyte differentiation. The assay was conducted using a human monocytic leukemia cell line, THP-1, treated with either 12-O-tetradecanoylphorbol 13-acetate or cytokines, such as tumor necrosis factor alpha and interferon-gamma. This method is sensitive and easy to perform, especially in case of analyzing a large number of samples. PMID- 17920621 TI - Laboratory trials to infect insects and nematodes by some acaropathogenic Hirsutella strains (Mycota: Clavicipitaceous anamorphs). AB - Laboratory assays have been carried out to artificially infect insect larvae of the birch bark-beetle (Scolytus ratzeburgi Jans.-Coleoptera, Scolytidae) and codling moth Cydia pomonella L. -Lepidoptera, Tortricidae) as well as the potato cyst nematode-Globodera rostochiensis Wollenweber, sugar beet nematode-Heterodera schachtii Schmidt and root-knot nematode-Meloidogyne hapla Chif (Nematoda, Heteroderidae), by the phialoconidia of some fungal species of the genus Hirsutella. From among four species tested on insects only H. nodulosa Petch infected about 20% of S. ratzeburgi larvae, whereas H. kirchneri (Rostrup) Minter, Brady et Hall, H. minnesotensis Chen, Liu et Chen, and H. rostrata Balazy et Wisniewski did not affect insect larvae. Only single eggs of the root-knot nematode were infected by H. minnesotensis in the laboratory trials, whereas its larvae remained unaffected. No infection cases of the potato cyst nematode (G. rostochiensis) and sugar beet nematode eggs were obtained. Comparisons of DNA-ITS region sequences of the investigated strains with GenBank data showed no differences between H. minnesotensis isolates from the nematodes Heterodera glycines Ichinohe and from tarsonemid mites (authors' isolate). A fragment of ITS 2 with the sequence characteristic only for H. minnesotensis was selected. Two cluster analyses indicated close similarity of this species to H. thompsonii as sister clades, but the latter appeared more heterogenous. Insect and mite pathogenic species H. nodulosa localizes close to specialized aphid pathogen H. aphidis, whereas the phytophagous mite pathogens H. kirchneri and H. gregis form a separate sister clade. Hirsutella rostrata does not show remarkable relations to the establishment of aforementioned groups. Interrelated considerations on the morphology, biology and DNA sequencing of investigated Hirsutella species state their identification more precisely and facilitate the establishment of systematic positions. PMID- 17920622 TI - Adenosine A(1) receptor mediates delayed cardioprotective effect of sildenafil in mouse. AB - Sildenafil induces powerful cardioprotection against ischemia/reperfusion (I/R) injury. Since adenosine is known to be a major trigger of ischemic preconditioning, we hypothesized that A(1) adenosine receptor (A(1)AR) activation plays a role in sildenafil-induced cardioprotective signaling. Adult male C57BL wild-type (WT) mice or their corresponding A(1)AR knockout (A(1)AR-KO) mice were treated intraperitoneally (i.p.) with either sildenafil (0.71 mg/kg, equivalent to 50 mg dose for a 70 kg patient) or volume-matched saline. The selective A(1)AR antagonist 8-cyclopentyl-1,3-dipropyxanthine (DPCPX; 0.1 mg/kg, i.p.) was administered 30 min before sildenafil. The hearts were isolated 24 h later and subjected to 30 min of global ischemia and 1 h of reperfusion in Langendorff mode. Post-ischemic myocardial infarct size (mean+/-SEM; % of risk area) was reduced in C57BL-WT mice treated with sildenafil (5.6+/-0.9) versus saline control group (27.3+/-2.1; p<0.05; n=6/each). However, sildenafil failed to protect the A(1)AR-KO hearts (31.6+/-1.9 vs. 32.3+/-1.5 with saline, p>0.05). Additionally, DPCPX treatment abolished the infarct limiting effect of sildenafil (27.3+/-3.2, p<0.05). DPCPX alone had no effect on infarct size as compared with the control group. No significant changes in post-ischemic recovery of left ventricular pressure and heart rate were observed in the sildenafil-treated group. We further examined the effect of sildenafil in protection against simulated ischemia and reoxygenation injury in adult cardiomyocytes derived from WT and A(1)AR-KO mice. WT myocytes treated with sildenafil (1 microM) demonstrated significantly lower trypan blue-positive necrotic cells. However, cardiomyocytes derived from A(1)AR-KO mice or DPCPX-treated WT cells failed to show protection against necrosis with sildenafil. These results suggest that A(1)AR activation following treatment with sildenafil plays an integral role in the signaling cascade responsible for delayed protection against global I/R injury. PMID- 17920623 TI - Homodimeric structure and double-stranded RNA cleavage activity of the C-terminal RNase III domain of human dicer. AB - Human Dicer contains two RNase III domains (RNase IIIa and RNase IIIb) that are responsible for the production of short interfering RNAs and microRNAs. These small RNAs induce gene silencing known as RNA interference. Here, we report the crystal structure of the C-terminal RNase III domain (RNase IIIb) of human Dicer at 2.0 A resolution. The structure revealed that the RNase IIIb domain can form a tightly associated homodimer, which is similar to the dimers of the bacterial RNase III domains and the two RNase III domains of Giardia Dicer. Biochemical analysis showed that the RNase IIIb homodimer can cleave double-stranded RNAs (dsRNAs), and generate short dsRNAs with 2 nt 3' overhang, which is characteristic of RNase III products. The RNase IIIb domain contained two magnesium ions per monomer around the active site. The distance between two Mg-1 ions is approximately 20.6 A, almost identical with those observed in bacterial RNase III enzymes and Giardia Dicer, while the locations of two Mg-2 ions were not conserved at all. We presume that Mg-1 ions act as catalysts for dsRNA cleavage, while Mg-2 ions are involved in RNA binding. PMID- 17920625 TI - Human cataract lens membrane at subnanometer resolution. AB - Human pathologies often originate from molecular disorders. Therefore, imaging technology as one of the bases for the identification and understanding of pathologies must provide views of single molecules at subnanometer resolution. Membrane proteins mediate many of life's most important processes, and their malfunction is often lethal or leads to severe disease. The membrane proteins aquaporin-0 (AQP0) and connexons form junctional microdomains between healthy lens core cells in which AQP0 form square arrays surrounded by connexons. Malfunction of both proteins results in the formation of cataract. We have used high-resolution atomic force microscopy (AFM) to image junctional microdomains in membranes from an individual human eye lens with senile cataract. Images at subnanometer resolution report individual helix-connecting loops of four amino acid residues on the AQP0 surface. We describe the supramolecular assembly and the conformational state of AQP0 in junctional microdomains, where a mixture of truncated junctional and full-length water channel AQP0 form square arrays. Imaging of microdomain borders revealed individual AQP0 tetramers and no associated connexon, indicating a lack of metabolite transport, waste accumulation, and enlarged regions of non-adhering membranes, causing cataract in this individual. This first high-resolution view of the membrane of this pathological human tissue provides insights into cataract pathology at the single membrane protein level, and indicates the power of the AFM as a future tool in medical imaging at subnanometer resolution. PMID- 17920624 TI - Electrostatic contributions to the stability of the GCN4 leucine zipper structure. AB - Ion pairs are ubiquitous in X-ray structures of coiled coils, and mutagenesis of charged residues can result in large stability losses. By contrast, pK(a) values determined by NMR in solution often predict only small contributions to stability from charge interactions. To help reconcile these results we used triple resonance NMR to determine pK(a) values for all groups that ionize between pH 1 and 13 in the 33 residue leucine zipper fragment, GCN4p. In addition to the native state we also determined comprehensive pK(a) values for two models of the GCN4p denatured state: the protein in 6 M urea, and unfolded peptide fragments of the protein in water. Only residues that form ion pairs in multiple X-ray structures of GCN4p gave large pK(a) differences between the native and denatured states. Moreover, electrostatic contributions to stability were not equivalent for oppositely charged partners in ion pairs, suggesting that the interactions between a charge and its environment are as important as those within the ion pair. The pH dependence of protein stability calculated from NMR-derived pK(a) values agreed with the stability profile measured from equilibrium urea-unfolding experiments as a function of pH. The stability profile was also reproduced with structure-based continuum electrostatic calculations, although contributions to stability were overestimated at the extremes of pH. We consider potential sources of errors in the calculations, and how pK(a) predictions could be improved. Our results show that although hydrophobic packing and hydrogen bonding have dominant roles, electrostatic interactions also make significant contributions to the stability of the coiled coil. PMID- 17920626 TI - An allosteric rheostat in HIV-1 gp120 reduces CCR5 stoichiometry required for membrane fusion and overcomes diverse entry limitations. AB - Binding of the human immunodeficiency virus (HIV-1) envelope glycoprotein gp120 to the CCR5 co-receptor reduces constraints on the metastable transmembrane subunit gp41, thereby enabling gp41 refolding, fusion of viral and cellular membranes, and infection. We previously isolated adapted HIV-1(JRCSF) variants that more efficiently use mutant CCR5s, including CCR5(Delta18) lacking the important tyrosine sulfate-containing amino terminus. Effects of mutant CCR5 concentrations on HIV-1 infectivities were highly cooperative, implying that several may be required. However, because wild-type CCR5 efficiently mediates infections at trace concentrations that were difficult to measure accurately, analyses of its cooperativity were not feasible. New HIV-1(JRCSF) variants efficiently use CCR5(HHMH), a chimera containing murine extracellular loop 2. The adapted virus induces large syncytia in cells containing either wild-type or mutant CCR5s and has multiple gp120 mutations that occurred independently in CCR5(Delta18)-adapted virus. Accordingly, these variants interchangeably use CCR5(HHMH) or CCR5(Delta18). Additional analyses strongly support a novel energetic model for allosteric proteins, implying that the adaptive mutations reduce quaternary constraints holding gp41, thus lowering the activation energy barrier for membrane fusion without affecting bonds to specific CCR5 sites. In accordance with this mechanism, highly adapted HIV-1s require only one associated CCR5(HHMH), whereas poorly adapted viruses require several. However, because they are allosteric ensembles, complexes with additional co-receptors fuse more rapidly and efficiently than minimal ones. Similarly, wild-type HIV-1(JRCSF) is highly adapted to wild-type CCR5 and minimally requires one. The adaptive mutations cause resistances to diverse entry inhibitors and cluster appropriately in the gp120 trimer interface overlying gp41. We conclude that membrane fusion complexes are allosteric machines with an ensemble of compositions, and that HIV 1 adapts to entry limitations by gp120 mutations that reduce its allosteric hold on gp41. These results provide an important foundation for understanding the mechanisms that control membrane fusion and HIV-1's facile adaptability. PMID- 17920627 TI - Promiscuous stimulation of ParF protein polymerization by heterogeneous centromere binding factors. AB - The segrosome is the nucleoprotein complex that mediates accurate segregation of bacterial plasmids. The segrosome of plasmid TP228 comprises ParF and ParG proteins that assemble on the parH centromere. ParF, which exemplifies one clade of the ubiquitous ParA superfamily of segregation proteins, polymerizes extensively in response to ATP binding. Polymerization is modulated by the ParG centromere binding factor (CBF). The segrosomes of plasmids pTAR, pVT745 and pB171 include ParA homologues of the ParF subgroup, as well as diverse homodimeric CBFs with no primary sequence similarity to ParG, or each other. Centromere binding by these analogues is largely specific. Here, we establish that the ParF homologues of pTAR and pB171 filament modestly with ATP, and that nucleotide hydrolysis is not required for this polymerization, which is more prodigious when the cognate CBF is also present. By contrast, the ParF homologue of plasmid pVT745 did not respond appreciably to ATP alone, but polymerized extensively in the presence of both its cognate CBF and ATP. The co-factors also stimulated nucleotide-independent polymerization of cognate ParF proteins. Moreover, apart from the CBF of pTAR, the disparate ParG analogues promoted polymerization of non-cognate ParF proteins suggesting that filamentation of the ParF proteins is enhanced by a common mechanism. Like ParG, the co-factors may be modular, possessing a centromere-specific interaction domain linked to a flexible region containing determinants that promiscuously stimulate ParF polymerization. The CBFs appear to function as bacterial analogues of formins, microtubule associated proteins or related ancillary factors that regulate eucaryotic cytoskeletal dynamics. PMID- 17920628 TI - Allosteric loss-of-function mutations in HIV-1 Nef from a long-term non progressor. AB - Activation of Src family kinases by human immunodeficiency virus type 1 (HIV-1) Nef may play an important role in the pathogenesis of HIV/AIDS. Here we investigated whether diverse Nef sequences universally activate Hck, a Src family member expressed in macrophages and other HIV-1 target cells. In general, we observed that Hck activation is a highly conserved Nef function. However, we identified an unusual Nef variant from an HIV-positive individual that did not develop AIDS which failed to activate Hck despite the presence of conserved residues linked to Hck SH3 domain binding and kinase activation. Amino acid sequence alignment with active Nef proteins revealed differences in regions not previously implicated in Hck activation, including a large internal flexible loop absent from available Nef structures. Substitution of these residues in active Nef compromised Hck activation without affecting SH3 domain binding. These findings show that residues at a distance from the SH3 domain binding site influence Nef interactions allosterically with a key effector protein linked to AIDS progression. PMID- 17920630 TI - Divergent genetic control of protein solubility and conformational quality in Escherichia coli. AB - In bacteria, protein overproduction results in the formation of inclusion bodies, sized protein aggregates showing amyloid-like properties such as seeding-driven formation, amyloid-tropic dye binding, intermolecular beta-sheet architecture and cytotoxicity on mammalian cells. During protein deposition, exposed hydrophobic patches force intermolecular clustering and aggregation but these aggregation determinants coexist with properly folded stretches, exhibiting native-like secondary structure. Several reports indicate that inclusion bodies formed by different enzymes or fluorescent proteins show detectable biological activity. By using an engineered green fluorescent protein as reporter we have examined how the cell quality control distributes such active but misfolded protein species between the soluble and insoluble cell fractions and how aggregation determinants act in cells deficient in quality control functions. Most of the tested genetic deficiencies in different cytosolic chaperones and proteases (affecting DnaK, GroEL, GroES, ClpB, ClpP and Lon at different extents) resulted in much less soluble but unexpectedly more fluorescent polypeptides. The enrichment of aggregates with fluorescent species results from a dramatic inhibition of ClpP and Lon-mediated, DnaK-surveyed green fluorescent protein degradation, and it does not perturb the amyloid-like architecture of inclusion bodies. Therefore, the Escherichia coli quality control system promotes protein solubility instead of conformational quality through an overcommitted proteolysis of aggregation prone polypeptides, irrespective of their global conformational status and biological properties. PMID- 17920629 TI - Bis-methionyl coordination in the crystal structure of the heme-binding domain of the streptococcal cell surface protein Shp. AB - Surface proteins Shr, Shp, and the ATP-binding cassette (ABC) transporter HtsABC are believed to make up the machinery for heme uptake in Streptococcus pyogenes. Shp transfers its heme to HtsA, the lipoprotein component of HtsABC, providing the only experimentally demonstrated example of direct heme transfer from a surface protein to an ABC transporter in Gram-positive bacteria. To understand the structural basis of heme transfer in this system, the heme-binding domain of Shp (Shp(180)) was crystallized, and its structure determined to a resolution of 2.1 A. Shp(180) exhibits an immunoglobulin-like beta-sandwich fold that has been recently found in other pathogenic bacterial cell surface heme-binding proteins, suggesting that the mechanisms of heme acquisition are conserved. Shp shows minimal amino acid sequence identity to these heme-binding proteins and the structure of Shp(180) reveals a unique heme-iron coordination with the axial ligands being two methionine residues from the same Shp molecule. A negative electrostatic surface of protein structure surrounding the heme pocket may serve as a docking interface for heme transfer from the more basic outer cell wall heme receptor protein Shr. The crystal structure of Shp(180) reveals two exogenous, weakly bound hemins, which form a large interface between the two Shp(180) molecules in the asymmetric unit. These "extra" hemins form a stacked pair with a structure similar to that observed previously for free hemin dimers in aqueous solution. The propionates of the protein-bound heme coordinate to the iron atoms of the exogenous hemin dimer, contributing to the stability of the protein interface. Gel filtration and analytical ultracentrifugation studies indicate that both full-length Shp and Shp(180) are monomeric in dilute aqueous solution. PMID- 17920631 TI - Human embryonic stem cells: an experimental and therapeutic resource for neurological disease. AB - There is a great and unmet need for meaningful therapies that will deliver restorative solutions to patients with neurological disorders such as multiple sclerosis (MS), Parkinson's disease and stroke. The emergence of human embryonic stem cells as an experimental and therapeutic resource represents a major opportunity for brain repair. Embryonic stem cells offer the potential to study human cells, model disease, accelerate drug discovery and of themselves act as a cell-based therapy. In contrast to other organs, a "one size fits all" approach is inappropriate for repair of the brain; rather therapies need to be "bespoke". The design and development of embryonic stem-cell based CNS reparative strategies pose many challenges, both conceptual and practical. Using multiple sclerosis as an example, this paper addresses the needs for the translation of embryonic stem cell biology to regenerative neurology. PMID- 17920632 TI - Origins of blood volume change due to glutamatergic synaptic activity at astrocytes abutting on arteriolar smooth muscle cells. AB - The cellular mechanisms that couple activity of glutamatergic synapses with changes in blood flow, measured by a variety of techniques including the BOLD signal, have not previously been modelled. Here we provide such a model, that successfully accounts for the main observed changes in blood flow in both visual cortex and somatosensory cortex following their stimulation by high-contrast drifting grating or by single whisker stimulation, respectively. Coupling from glutamatergic synapses to smooth muscle cells of arterioles is effected by astrocytes releasing epoxyeicosatrienoic acids (EETs) onto them, following glutamate stimulation of the astrocyte. Coupling of EETs to the smooth muscle of arterioles is by means of potassium channels in their membranes, leading to hyperpolarization, relaxation and hence an increase in blood flow. This model predicts a linear increase in blood flow with increasing numbers of activated astrocytes, but a non-linear increase with increasing glutamate release. PMID- 17920633 TI - Factors influencing route choice by avian migrants: a dynamic programming model of Pacific brant migration. AB - We used stochastic dynamic programming to investigate a spectacular migration strategy in the black brant Branta bernicla nigricans, a species of goose. Black brant migration is well suited for theoretical analysis since there are a number of existing strategies that easily can be compared. In early autumn, almost the entire population of the black brant gathers at Izembek Lagoon on the Alaska Peninsula to stage and refuel before the southward migration. There are at least three distinct strategies, with most geese making a spectacular direct migration more than 5000km across the Gulf of Alaska to their wintering grounds in southern Baja California or mainland Mexico. This is a potentially dangerous strategy since foraging is not possible during the overseas passage. Some individuals instead use shorter flights to make a detour along the coast, a longer route that all individuals use for northwards migration in spring. Since flight costs accelerate with increasing body mass, migration by short flights is energetically cheaper than long-distance flights. A small but increasing part of the population has recently begun to winter at Izembek. We investigated this migration under two different suppositions using a dynamic state variable model. First, if the geese are free to make a strategic choice, under what assumptions should they prefer direct migration and under what assumptions should they prefer detour migration/winter residency? Second, provided that the dominating direct migration strategy is optimal, what conditions will force the geese to go for detour migration/winter residency? In the second case the geese may try to follow an optimal direct migration strategy, but stochastic events may force them to choose a suboptimal policy. We also simulated possible effects of global warming. The model suggests that the fuel level at arrival in Izembek and fuel gain rates are key factors and that tail winds must have been reliable in the past, otherwise direct migration could not have evolved. It also suggests that a change to milder winters may promote an unexpectedly abrupt change from long-distance to short distance migration or winter residency. Finally, it produced a number of predictions that might be testable in the field. PMID- 17920634 TI - Temperature dependence and temperature compensation of kinetics of chemical oscillations; Belousov-Zhabotinskii reaction, glycolysis and circadian rhythms. AB - Based on the distribution of activation energies around the experimental mean and averaging of rate constants we propose a theoretical scheme to examine the temperature dependence and temperature compensation of time periods of chemical oscillations. The critical finite width of the distribution is characteristic of endogeneous oscillations for compensating kinetics as observed in circadian oscillations, while the vanishing width corresponds to Arrhenius temperature dependent kinetics of non-endogeneous chemical oscillation in Belousov Zhabotinskii reaction in a CSTR or glycolysis in cell-free yeast extracts. Our theoretical analysis is corroborated with experimental data. PMID- 17920635 TI - Minimizing invader impacts: striking the right balance between removal and restoration. AB - Invasive species can cause severe damage in their introduced range; this damage often persists even after removal of the invader. In order to efficiently allocate a limited budget between invader removal and restoration of habitat from which the invader has been removed, it is vital to quantify the impacts of the invasion within an economic context. Here we develop optimal management strategies for biological invasions, which minimize both the direct economic costs of removal and restoration, and the ecological costs of present and future damage caused by the invasion. We demonstrate how this can be formulated as a linear programming problem, enabling the fast and efficient computation of optimal solutions. Using a simple example, we outline some general principles for the optimal control of an invader that damages its environment. Notably, we show that the most effective strategies often switch the priority of removal and restoration over time, and outline the conditions under which restoration is prioritized over removal. The proportion of total funds allocated to restoration will depend on the annual budget, the persistence of damage, and the relative costs of damage, removal and restoration. PMID- 17920636 TI - Glucose-regulated insulin production from genetically engineered human non-beta cells. AB - In this report we describe development and characterization of four human cell lines that are able to secrete insulin and C-peptide in response to higher concentrations of glucose. These cell lines have been developed by stably and constitutively expressing human proinsulin with a furin-cleavable site, whereas expression of furin is regulated by glucose concentration. These cell lines have been cloned and, therefore, the transgene in each cell is located in an identical location of the genome leading to a uniform expression. Cloning has also allowed us to identify cell lines with more desirable properties such as higher basal insulin secretion and/or better glucose responsiveness. We have further shown that the insulin produced by these cells is biologically active and induces normoglycemia when injected in diabetic animals. Our objective in initiating these studies was to identify a cell line that could serve as a surrogate beta cell line for therapeutic intervention in type I diabetic patients. PMID- 17920637 TI - Increased susceptibility of small intestine to NSAID-provoked ulceration in rats with adjuvant-induced arthritis: involvement of enhanced expression of TLR4. AB - NSAIDs damage the small intestine as well as the stomach as adverse effects. We previously reported that the gastric ulcerogenic response to NSAIDs was markedly increased in arthritic rats. The present study was designed to examine the intestinal ulcerogenic property of indomethacin in adjuvant-induced arthritic rats in comparison with normal animals. Arthritis was induced in male Dark Agouti rats by injection of Freund's complete adjuvant into the right hindfoot. Two weeks later, indomethacin was given orally and the intestine was examined for lesions at several time points after indomethacin. Indomethacin produced intestinal lesions in both normal and arthritic rats, but in the latter, the ulcerogenic response occurred much earlier and the severity was markedly enhanced. Aminoguanidine, an inhibitor of iNOS, significantly suppressed the damage, yet the efficacy differed in normal and arthritic rats, depending on the dose schedule; the effect of post-administration (6 h after) was greater than that of pre-administration (0.5 h before) in normal rats, whereas that of post administration was less than that of pre-administration in arthritic rats. The expression of iNOS and TLR4 in the intestine was enhanced in arthritic rats as compared with normal rats. These results suggest that the intestinal ulcerogenic response to indomethacin is markedly aggravated in arthritic rats. Notably, the onset of the ulceration was much earlier in arthritic rats than normal rats. These phenomena may be accounted for by the upregulation of iNOS/NO through the increased expression of TLR4 in the small intestine of arthritic rats. PMID- 17920638 TI - Concentration-dependent vasoconstrictive effect of hyperoxia on hypercarbia dilated retinal arterioles. AB - BACKGROUND/AIMS: The relative effects of simultaneously administered oxygen and carbon dioxide on vascular resistance are unknown. The purpose of the study was to investigate the independent effect of oxygen partial pressure on hypercarbia induced vasodilation in the retinal arterioles. METHODS: Twelve young healthy volunteers participated in the study. End-tidal partial pressure of carbon dioxide was raised 23% from the baseline (i.e. air) at normoxia and then maintained constant while end-tidal partial pressure of oxygen (PETO(2)) was raised in a stepwise incremental fashion. Retinal vessel diameter and blood velocity were measured in the superior-temporal arteriole using the Canon Laser Blood Flowmeter. RESULTS: Hypercarbia resulted in a 16% increase in blood velocity and a 22% increase in blood flow (p<0.05). At maximal hyperoxia (group mean PETO(2) of 556 mm Hg) vessel diameter, blood velocity and flow were reduced by 9%, 22% and 36%, respectively, relative to baseline (p<0.001). CONCLUSION: The concentration-dependent vasoconstrictive effect of oxygen in retinal arterioles was quantified for the first time by implementing precise control of end-tidal concentrations of CO(2) and O(2). Oxygen-induced vasoconstriction is sufficiently potent to offset and reverse hypercarbia-induced vasodilation. PMID- 17920639 TI - Impaired local microvascular vasodilatory effects of insulin and reduced skin microvascular vasomotion in obese women. AB - Our study aim is to investigate whether obesity is characterized by an impairment of insulin-mediated vasodilatory effects and by a modification of basal vasomotion in the skin microvasculature. Forty healthy obese and forty healthy lean women were included. Microvascular effects of insulin as compared to a control substance were measured by cathodal iontophoresis combined with laser Doppler flowmetry. Vasomotion was examined by Fourier transform analyses of skin laser Doppler flow at rest. Locally administered insulin, as compared to the control substance, induced a microvascular vasodilatory response in lean (median (interquartile range): 31.6 (17.1-43.9) vs. 22.9 (16.4-36.7) perfusion units, P=0.04), but not in obese women (28.1 (14.4-47.1) vs. 27.5 (17.5-48.2) perfusion units, P=0.7). The relative insulin-induced increase in blood flow corrected for the control substance was higher in lean than obese women (ANOVA for repeated measures F=3.93, P=0.05). The contribution of the total frequency spectrum 0.01 1.6 Hz and of the frequency intervals 0.01-0.02 Hz and 0.02-0.06 Hz (representative of endothelial and neurogenic activity, respectively) to basal microvascular vasomotion was lower in obese than in lean women (P<0.05 for all). These findings show that obesity is characterized by an impaired direct microvascular vasodilatory effect of insulin and by decreased skin microvascular vasomotion in a way that is suggestive for alterations of endothelial and neurogenic activity. PMID- 17920640 TI - The buccal cytome and micronucleus frequency is substantially altered in Down's syndrome and normal ageing compared to young healthy controls. AB - The buccal micronucleus cytome assay was used to investigate biomarkers for DNA damage, cell death and basal cell frequency in buccal cells of healthy young, healthy old and young Down's syndrome cohorts. With normal ageing a significant increase in cells with micronuclei (P<0.05, average increase +366%), karyorrhectic cells (P<0.001, average increase +439%), condensed chromatin cells (P<0.01, average increase +45.8%) and basal cells (P<0.001, average increase +233%) is reported relative to young controls. In Down's syndrome we report a significant increase in cells with micronuclei (P<0.001, average increase +733%) and binucleated cells (P<0.001, average increase +84.5%) and a significant decrease in condensed chromatin cells (P<0.01, average decrease -52%), karyolytic cells (P<0.001, average decrease -51.8%) and pyknotic cells (P<0.001, average decrease -75.0%) relative to young controls. These changes show distinct differences between the cytome profile of normal ageing relative to that for a premature ageing syndrome, and highlight the diagnostic value of the cytome approach for measuring the profile of cells with DNA damage, cell death and proportion of cells with proliferative potential (i.e., basal cells). Significant correlations amongst cell death biomarkers observed in this study were used to propose a new model of the inter-relationship of cell types scored within the buccal micronucleus cytome assay. This study validates the use of a cytome approach to investigate DNA damage, cell death and cell proliferation in buccal cells with ageing. PMID- 17920641 TI - Robotic movement elicits visuomotor priming in children with autism. AB - The ability to understand another person's action and, if needed, to imitate that action, is a core component of human social behaviour. Imitation skills have attracted particular attention in the search for the underlying causes of the social difficulties that characterize autism. In recent years, it has been reported that people with autism can bypass some of their social deficits by interacting with robots. However, the robot preference in terms of imitation has yet to be proved. Here we provide empirical evidence that interaction with robots can trigger imitative behaviour in children with autism. We compared a group of high functioning children with autism with a group of typically developing children in a visuomotor priming experiment. Participants were requested to observe either a human or a robotic arm model performing a reach-to-grasp action towards a spherical object. Subsequently, the observers were asked to perform the same action towards the same object. Two 'control' conditions in which participants performed the movement in the presence of either the static human or robot model were also included. Kinematic analysis was conducted on the reach-to grasp action performed by the observer. Our results show that children with autism were facilitated - as revealed by a faster movement duration and an anticipated peak velocity - when primed by a robotic but not by a human arm movement. The opposite pattern was found for normal children. The present results show that interaction with robots has an effect on visuomotor priming processes. These findings suggest that in children with autism the neural mechanism underlying the coding of observed actions might be tailored to process socially simpler stimuli. PMID- 17920642 TI - Avoidance of emotionally arousing stimuli predicts social-perceptual impairment in Asperger's syndrome. AB - We combined eye-tracking technology with a test of facial affect recognition and a measure of self-reported social anxiety in order to explore the aetiology of social-perceptual deficits in Asperger's syndrome (AS). Compared to controls matched for age, IQ and visual-perceptual ability, we found a group of AS adults was impaired in their recognition of fearful and sad expressions and spent significantly less time fixating the eye region of all faces. For AS subjects, but not controls, the extent of the failure to fixate the eyes predicted the degree of impairment at recognising fearful expressions. In addition, poor fear recognition and reduced fixation of the eyes were independently associated with greater levels of social anxiety in AS individuals. These findings support the hypothesis that avoidance of emotionally arousing stimuli, such as eyes, contributes to social-perceptual impairment in AS. Furthermore, our findings are consistent with theories implicating amygdala-mediated over-arousal and anxiety in the development of these social-perceptual deficits. PMID- 17920643 TI - Hue-specific colour memory impairment in an individual with intact colour perception and colour naming. AB - Cases of hue-selective dyschomatopsias, together with the results of recent optical imaging studies [Xiao, Y., Casti, A. R. R., Xiao, J., & Kaplan, E. (2006). A spatially organized representation of colour in macaque primary visual cortex. Perception, 35, ECVP Abstract Supplement; Xiao, Y., Wang, Y., & Felleman, D. J. (2003). A spatially organized representation of colour in macaque cortical area V2. Nature, 421, 535-539], have provided support for the idea that different colours are processed in spatially distinct regions of extrastriate cortex. In the present report, we provide evidence suggesting that a similar, but distinct, map may exist for representations of colour in memory. This evidence comes from observations of a young woman (QP) who demonstrates an isolated deficit in colour memory secondary to a concussive episode. Despite having normal colour perception and colour naming skills, and above-average memory skills in other domains, QP's ability to recall visually encoded colour information over short retention intervals is dramatically impaired. Her long-term memory for colour and her colour imagery skills are also abnormal. Surprisingly, however, these impairments are not seen with all hues; specifically, her ability to remember or imagine blue shades is spared. This interesting case contributes to the literature suggesting that colour perception, naming, and memory can be clinically dissociated, and provides insights into the organization of colour information in memory. PMID- 17920644 TI - Effects of REM deprivation and an NMDA agonist on the extinction of conditioned fear. AB - Rapid eye movement sleep (REM) has been implicated in a number of learning and memory tasks. Previous research has demonstrated that REM deprivation impairs the development of extinction of conditioned fear responses. However, the neurobiological mechanisms of this effect remain unclear. The present study investigated the effects of systemic administration of d-cycloserine (DCS), an NMDA agonist, on the extinction of a conditioned fear response following 6 h of REM deprivation. In experiment 1, rats were administered DCS between fear training and REM deprivation. In experiment 2, rats were administered DCS prior to extinction training. The results of experiment 1 indicated that both DCS alone and REM deprivation alone impaired extinction learning. Administration of DCS to REM deprived animals partially, but not completely, reversed the deficit in extinction. The results of experiment 2 indicated that regardless of prior REM deprivation history, DCS facilitated extinction learning. The results provide further evidence for a role of REM in the extinction of cued fear learning and indicate that this effect appears to be partially mediated by NMDA-dependent mechanisms. PMID- 17920645 TI - Weakened [corrected] taste-LPS association during endotoxin tolerance. AB - In naive individuals, the administration of bacterial lipopolysaccharide (LPS) provokes a rapid systemic increase in pro-inflammatory cytokines such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6, inducing an acute phase response including sickness behavior. Strong associative learning occurs when relevant gustatory/olfactory stimuli precede the activation of the immune system, affecting long-term individual food selection and nutritional strategies. Repeated LPS administration results in the development of an endotoxin tolerance status, characterized by a drastic reduction in the LPS-induced cytokine response. Here we investigated how the postprandial categorization of a relevant taste (0.2% saccharin) changed after administration of a high dose of LPS (0.5 mg/kg i.p.) in LPS-tolerant animals. Determination of the consummatory fluid intake revealed that, in contrast to LPS-naive rats, taste-LPS association did not occur during endotoxin tolerance. Ninety minutes after the single association trial, the plasma responses of TNF-alpha, IL-1beta and IL-6 were completely blunted in LPS-tolerant animals, which also resulted in low LPS-adipsogenic and LPS-anorexic effects. These findings indicate that an identical immune challenge can result in completely different neuro-behavioral consequences depending on the immune history of the individual, thus revealing part of the complex interconnection between the immune and neuro-endocrine systems in regulating food selection and consumption during the infectious process. PMID- 17920646 TI - The effect of season and weather on physical activity: a systematic review. AB - OBJECTIVES: This study reviewed previous studies to explore the effect of season, and consequently weather, on levels of physical activity. STUDY DESIGN AND METHODS: Thirty-seven primary studies (published 1980-2006) representing a total of 291883 participants (140482 male and 152085 female) from eight different countries are described, and the effect of season on moderate levels of physical activity is considered. RESULTS: Upon review of the evidence, it appears that levels of physical activity vary with seasonality, and the ensuing effect of poor or extreme weather has been identified as a barrier to participation in physical activity among various populations. Therefore, previous studies that did not recognize the effect of weather and season on physical activity may, in fact, be poor representations of this behaviour. CONCLUSIONS: Future physical activity interventions should consider how weather promotes or hinders such behaviour. Providing indoor opportunities during the cold and wet months may foster regular physical activity behaviours year round. PMID- 17920647 TI - Plasma metabolite concentrations and hepatic enzyme activities in pregnant Romney ewes with restricted feeding. AB - Plasma metabolite concentrations and activities of enzymes related to energy metabolism in plasma, peripheral leukocytes and liver of pregnant Romney ewes with restricted feeding were measured to assess those metabolites and enzymes as indicators for evaluating metabolic conditions in the ewes. The body weights and plasma lactate concentrations of the low-feeding ewes (about 1.0 times maintenance) were significantly lower than those of the high-feeding ewes (about 1.5 times maintenance). There were no significant differences in plasma protein, glucose, triglyceride, cholesterol, pyruvate and immunoreactive insulin concentrations and plasma and leukocyte enzyme activities between both groups. Hepatic malate dehydrogenase activities were significantly higher and hexokinase activities were significantly lower in the low-feeding ewes than in the high feeding ewes. Restricted feeding could maintain pregnancy and some plasma metabolites and peripheral leukocyte enzymes may be useful indicators for evaluating metabolic changes in ewes. PMID- 17920648 TI - Taenia solium cysticercosis in Bali, Indonesia: serology and mtDNA analysis. AB - An active Taenia solium cysticercosis case in Bali, Indonesia, was followed-up by serology and computed tomography. Serology using semi-purified glycoprotein and recombinant antigens showed a drastic drop in titers after calcification of the cysts. Three paraffin-embedded cysts, prepared for histopathological examination, from three other patients were used for mtDNA analysis. The sequences of cox1 gene from T. solium cysticerci from Bali differed from those in Papua and other Asian countries. PMID- 17920649 TI - Characterization of Tityus scorpion venoms using synaptosome binding assays and reactivity towards Venezuelan and Brazilian antivenoms. AB - Venoms from Tityus species inhabiting five endemic regions of scorpionism in Venezuela (Andean, Perija range, north-central, northeastern, and Guayana) and also southeast Brazil (T. serrulatus and T. bahiensis) were characterized immunologically in ELISA experiments using mouse- and rabbit-derived antibodies to evaluate their cross-reactivity and also functionally, utilizing synaptosome binding assays. While Brazilian and Venezuelan antivenoms cross-reacted poorly, T. discrepans (north-central Venezuela) and T. zulianus (Andean) venoms shared a greater immunological relatedness than with T. perijanensis (Perija range). Anti T. breweri (Guayana) antibodies fully cross-reacted with T. discrepans. Native PAGE indicated species-specific fingerprints for all venoms and revealed differences between two populations (Anzoategui and Monagas States) of T. nororientalis (northeastern Venezuela). Components antigenically related to T. serrulatus beta-toxin TsVII were also detected in T. breweri, T. nororientalis (Anzoategui) and T. funestus (Andean). Antibodies against T. serrulatus anatoxin TsNTxP did not cross-react significantly with any Venezuelan venoms indicating lack of TsNTxP homologues. The results suggest that the extent of antigenic reactivity depends on the studied species rather than the geographical distance between their habitats. All venoms, with T. discrepans to a lesser extent, were able to significantly displace [(125)I]-TsVII from its binding site in rat brain synaptosomes. Our data indicate that beta-toxins functionally related to TsVII but differing significantly in their antigenic regions exist in Venezuelan venoms from different endemic regions. Identification of shared epitopes with TsVII, at least for some species, may lead to the design of antibodies based on common epitopes for treating scorpion envenoming in Venezuela and Brazil. PMID- 17920650 TI - Neutralizing monoclonal antibodies to human immunodeficiency virus type 1 do not inhibit viral transcytosis through mucosal epithelial cells. AB - HIV-1 transcytosis has been proposed as a potential mechanism allowing the virus to cross the epithelium during mucosal transmission. Epitopes of the HIV-1 envelope involved in this process have not been identified yet. Here, we assessed a large panel of HIV neutralizing antibodies recognizing well-characterized epitopes of the HIV-1 envelope for their ability to block HIV-1 transcytosis across a confluent epithelial monolayer. We found that all of the 13 HIV-1 specific monoclonal antibodies tested in the present study, including the three broadly neutralizing antibodies 2F5, 2G12 and IgG1b12, lacked the ability to inhibit transcytosis of cell-free and cell-associated R5- as X4-tropic HIV-1 across a tight and polarized monolayer of HEC-1 epithelial cells. In contrast, anti-gp160 polyclonal antibodies purified from serum or breast milk of HIV-1 infected individuals potently inhibited HIV-1 transcytosis. Furthermore, polymeric S-IgA exhibited similar ability to inhibit transcytosis compared to IgG despite their lower anti-gp160 specific activity. Together, these results demonstrate that the major neutralizing envelope epitopes of HIV-1 are not involved in HIV-1 transcytosis, and suggest that surface agglutination of virus particles may participate to the blocking effect observed with both polyclonal and polymeric anti-gp160 immunoglobulins. PMID- 17920651 TI - A transgenic mouse model for gene therapy of rhodopsin-linked Retinitis Pigmentosa. AB - Mutational heterogeneity in genes causative of dominantly inherited disorders represents a significant barrier for development of therapies directed towards correction of the primary genetic defect. To circumvent the mutational heterogeneity present in rhodopsin- (RHO-) linked autosomal dominant Retinitis Pigmentosa (adRP), a strategy involving suppression and replacement of RHO has been adopted. RNA interference- (RNAi-) mediated suppression of RHO has been explored as has the generation of an RNAi-resistant replacement gene using the degeneracy of the genetic code. Additionally, the functional equivalence of codon modified replacement genes has been demonstrated in a transgenic animal (RHO-M). Suppression and replacement, while exemplified by adRP, may also be relevant to many other dominantly inherited diseases with the hallmark of mutational heterogeneity. PMID- 17920652 TI - Effects of chemical amendments on aquatic floc structure, settling and strength. AB - Using a shear-cell/flow-cell combination integrated with an inverted microscope, the behaviour of Hamilton Harbour sediments was studied mixed with three different amendments: alum, chitosan (both coagulants) and a polyacrylamide (a flocculant). Samples from the shear cell were drawn into the flow cell, where floc structure and size were assessed throughout the floc formation and breakage stages using computer image analysis. Settling velocity, density and porosity were also assessed, with results suggesting that amendment addition may be an effective method for the management of high-turbidity environments, provided there are no toxicological effects. In an assessment of performance, it was found that the polyacrylamide flocculant showed the greatest promise in reducing turbidity levels as it produced the largest flocs with the highest settling velocity. Although more prone to break-up, these flocs still remained larger than those formed with alum or chitosan at the same shear. All flocs, regardless of amendment, broke up due to a fracture mechanism rather than by microscale erosion. By improving our understanding of how these amendments may influence floc properties and behaviours, more effective management tools may be developed for the remediation and control of high-turbidity aquatic environments. PMID- 17920653 TI - Catalytic oxidation of gaseous PCDD/Fs with ozone over iron oxide catalysts. AB - Catalytic oxidation of PCDD/Fs (polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans) with ozone (catalytic ozonation) over nano-sized iron oxides (denoted as FexOy) was carried out at temperature of 120-180 degrees C. The effects of operating temperature, ozone concentration, space velocity (SV) and water vapor contents on PCDD/F removal and destruction efficiencies via catalytic ozonation were investigated. High activity of the iron oxide catalyst towards PCDD/F decomposition was observed even at low temperatures with the aid of ozone. The PCDD/F removal and destruction efficiencies achieved with FexOy/O3 at 180 degrees C reach 94% and 91%, respectively. In the absence of ozone, the destruction efficiencies of all PCDD/F congeners are below 20% and decrease with increasing chlorination level of PCDD/F congener at lower temperature (120 degrees C). However, in the presence of ozone, the destruction efficiencies of all PCDD/F congeners are over 80% on FexOy/O3 at 180 degrees C. Higher temperature and ozone addition increase the activity of iron oxide for the decomposition of PCDD/Fs. Additionally, in the presence of 5% water vapor, the destruction efficiency of the PCDD/Fs is above 90% even at lower operating temperature (150 degrees C). It indicates that the presence of appropriate amount of water vapor enhances the catalytic activity for the decomposition of gas-phase PCDD/Fs. PMID- 17920654 TI - Unusual arsenic speciation in sea anemones. AB - Nine species of sea anemones (Anthopleura asiatica, Actinia equina, Actinodendron arboreum, Phymanthus loligo, Entacmaea actinostoloides, Stichodactyla gigantea, S. haddoni, S. mertensii and Metridium senile) contained arsenic in the range of 1.6-7.0microg As g(-1) (wet mass basis). Irrespective of the species, water soluble arsenic compounds accounted for more than 80% of the total arsenic. Analysis of water-soluble arsenic compounds by LC/ESI-MS revealed that four arsenicals, arsenobetaine (AB), trimethylarsoniopropionate, arsenocholine (AC) and tetramethylarsonium ion (TEMA), are contained in most species but arsenate, methylarsonic acid, dimethylarsinic acid and trimethylarsine oxide are absent in all species. Interestingly, compositional patterns of the four arsenicals greatly differed from species to species. Only three species (S. gigantea, S. haddoni and M. senile) contained AB at the highest proportions, similar to the majority of marine animals. However, the remaining six species showed unusual compositional patterns of arsenic compounds; AC was most predominant in A. arboreum and P. loligo and TEMA in A. asiatica, A. equina, E. actinostoloides and S. mertensii. On the whole, high proportions (24.6-87.1% of the water-soluble arsenic) of TEMA appear to be a peculiar characteristic of many species of sea anemones. Thus, sea anemones are an important animal group in the arsenic cycling, especially in that they may be donors of TEMA to predators. PMID- 17920655 TI - Anaerobic transformation of polybrominated biphenyls with the goal of identifying unknown hexabromobiphenyls in Baltic cod liver. AB - Polybrominated biphenyls (PBBs) have been introduced as flame retardants in 1970. Despite decreasing application rates since the mid-1970s, PBB residues are still reported in the environment. Furthermore, environmental PBB residues often do not match the PBB pattern in technical products. To get insights into the structures of environmentally-relevant PBBs, the congener patterns of technical hexabromobiphenyl (THBB), octabromobiphenyl (TOBB), synthesized PBB 209 were compared to PBB residues in a cod liver sample from the Baltic Sea. The most relevant PBB congeners in Baltic cod liver were not present in the technical products and therefore most likely metabolites. For this reason, TOBB and HPLC fractions obtained from this technical product were incubated with super-reduced cyanocobalamine. Reductive debromination was found to be the predominant transformation process. Bromine substituents in ortho-positions proved to be more recalcitrant, and several of the unknown PBBs were tri- and tetra-ortho substituted congeners. Furthermore, the key-PBBs determined in Baltic cod liver were formed during this process. The most important hexabromobiphenyl in Baltic cod liver was identified as PBB 155 by parallel synthesis. PBB 155 which was not detected in the technical PBB product analyzed was suggested as an indicator PBB congener suited to decide whether PBB residues originate from the previous use of THBB (low relative abundance of PBB 155) or TOBB/TDBB (high relative abundance of PBB 155). The latter scenario was found to be valid for the Baltic cod liver sample. Thus, PBBs in the Baltic Sea appeared to originate from partially weathered residues of PBB 209. PMID- 17920656 TI - Prediction of CO maximum ground level concentrations in the Bay of Algeciras, Spain using artificial neural networks. AB - The region of the Bay of Algeciras is a very industrialized area where very few air pollution studies have been carried out. The main objective of this work has been the use of artificial neural networks (ANNs) as a predictive tool of high levels of ambient carbon monoxide (CO). Two approaches have been used: multilayer perceptron models (MLPs) with backpropagation learning rule and k-Nearest Neighbours (k-nn) classifiers, in order to predict future peaks of carbon monoxide. A resampling strategy with twofold cross-validation allowed the statistical comparison of the different topologies and models considered in the study. The procedure of random resampling permits an adequate and robust multiple comparisons of the tested models and allow us to select a group of best models. PMID- 17920657 TI - Survival of Escherichia coli O157:H7 in waters from lakes, rivers, puddles and animal-drinking troughs. AB - Survival of Escherichia coli O157:H7 in surface waters may increase the potential for dissipation of the organism to facilitate cycles of livestock re-infection and lead to human infection. Although previous studies have monitored survival of the organism in a range of surface waters, there is limited information on the influence of physico-chemical characteristics on persistence. Microcosms of four different surface water types (n=31) from the UK were inoculated with E. coli O157:H7 and incubated at 10 degrees C. The water types studied were: lake, puddle, river, and animal-drinking trough waters. Numbers of E. coli O157:H7 declined over time in all waters, although cells were still detected in 45% of non-sterile samples after 2 months. Persistence of E. coli O157:H7 was enhanced by water aeration and by prior sterilisation; however there was no correlation between water chemistry and mean E. coli O157:H7 die-off times or rates in any water type. Survival of the pathogen was better in lake and puddle waters than in river or drinking trough waters. Further studies are needed to establish the key water quality factors that regulate pathogen survival. PMID- 17920658 TI - Investigating and modelling the development of septic sewage in filled sewers under static conditions: a lab-scale feasibility study. AB - This paper describes a lab-scale study of the physical and bio-chemical processes associated with the development of septic conditions in sewer pipes filled with static sewage. The study has concentrated on the uptake of oxygen (OUR) and the subsequent changes in chemical oxygen demand (COD), sulphate, sulphide and nitrate concentration and the formation of volatile fatty acids (VFA). OUR of raw sewage ranged from 2 to 13 mg L(-1) h(-1). Apparent nitrate uptake and sulphide generation rates in static sewage varied between 0.2-0.7 mgNO(3) L(-1) h(-1) and 0.02-0.05 mgH(2)S-S L(-1) h(-1), respectively. A logistic function was used to simulate the sulphide generation process in static sewage. It was found that total COD (COD(total)) influenced the apparent sulphide generation rate while nitrate concentrations greater than 4 mg L(-1) controlled the onset of sulphide production in experiments without added sediment phase. Introducing a sediment phase appeared to accelerate hydrolysis and fermentation processes as evidenced by 5-14 times greater dissolved COD generation rates in the bulk water phase. PMID- 17920659 TI - Ochratoxin A production in relation to ecophysiological factors by Aspergillus section Nigri strains isolated from different substrates in Argentina. AB - Contamination of foodstuff with mycotoxins such as ochratoxins is a major matter of concern for human and animal health. In Aspergillus species, ochratoxin synthesis depends on several environmental factors. The aims of this work were to evaluate the effect of water activity (0.995-0.85), temperature (15, 25 and 30 degrees C), incubation time (7, 14 and 21 days) and their interactions on OTA production on peanut, maize kernels, dried grapes and coffee beans meal extract agar medium by eight strains of Aspergillus section Nigri isolated from human food in Argentina. The optimum temperature for OTA production was 25 or 30 degrees C depending on the strains assayed, in most cases the highest OTA levels were achieved after 7 days of incubation, whereas this situation occurred at 15 degrees C after 14 days of incubation for only one strain. The maximum OTA level was obtained at earlier growth states when incubation temperature increased. In general, OTA concentration increased as water activity (a(W)) increased with no significant production at 0.85-0.91 a(W) under all temperature levels tested. Production occurred over a range of temperatures (15-30 degrees C) with optimum production at 30 degrees C depending on a(W) assayed. The knowledge of Aspergillus section Nigri ecophysiology is critical in the development and prediction of the risk models of raw material and final product contamination by these species under fluctuating and interacting environmental parameters. PMID- 17920660 TI - Experimental investigation on the performance, gaseous and particulate emissions of a methanol fumigated diesel engine. AB - Experiments were conducted on a 4-cylinder direct-injection diesel engine with fumigation methanol injected into the air intake of each cylinder. The fumigation methanol was injected to top up 10%, 20% and 30% of the power output under different engine operating conditions. The effects of fumigation methanol on engine performance, gaseous emissions and particulate emission were investigated. The experimental results show that there is a decrease in the brake thermal efficiency when fumigation methanol is applied, except at the highest load of 0.67 MPa. At low loads, the brake thermal efficiency decreases with increase in fumigation methanol; but at high loads, it increases with increase in fumigation methanol. The fumigation method results in a significant increase in hydrocarbon (HC), carbon monoxide (CO), and nitrogen dioxide (NO(2)) emissions. The concentration of nitrogen oxides (NOx) is significantly reduced except at close to full load condition. There is also a reduction in the smoke opacity and the particulate matter (PM) mass concentration. For the submicron particles, the total number of particles decreases at low and medium loads but increases at high loads. In all cases, there is a shift of the particles towards smaller geometrical mean diameter, especially at high loads. The increase in nano-sized particles and the increase in NO(2) emission could have serious impact on human health. PMID- 17920661 TI - Suppression of HUVEC tissue factor synthesis by antisense oligodeoxynucleotide. AB - Tissue factor (TF) is an important regulator and effector molecule of coagulation. It is primary known as a cofactor for factor VIIa-mediated triggering of blood coagulation, which proceeds in a cascade of extracellular reactions, ultimately resulting in thrombin formation. In sepsis, expression of TF by activated monocytes, macrophages and endothelial cells may lead to disseminated intravascular coagulation. Further studies have suggested that TF also plays non-haemostatic roles in blood vessel development, tumor angiogenesis, metastasis and inflammation. In the present study we examined the feasibility of inhibiting lipopolysaccharide (LPS)-induced TF expression in cultured human umbilical vein endothelial cells (HUVECs) using a modified phosphorothioate antisense oligodeoxynucleotide targeted to the TF mRNA. CD31 receptor-mediated endocytosis was used as a means of delivering TF antisense oligomer to HUVECs. This DNA carrier system consists of anti-CD31 antibody conjugated to the antisense. Co-exposure of HUVECs with TF antisense and LPS resulted in 54.6+/ 3.2% suppression of TF activity when compared with control LPS stimulated cells. The antisense also reduced the LPS-induced TF mRNA level. Control experiments with TF sense and mismatched antisense oligomers were performed to exclude non specific inhibitory effects. The cytotoxicity of the antisense oligomer conjugate was also evaluated. Results demonstrate that this TF antisense oligomer specifically suppressed the synthesis of biologically active endothelial TF and that antisense oligomers might represent a useful tool in the investigation of endothelial TF function/biology. PMID- 17920662 TI - Gene expression profiling for the identification of G-protein coupled receptors in human platelets. AB - INTRODUCTION AND MATERIALS AND METHODS: G-protein coupled receptors (GPCRs) play an important role in platelet aggregation. To identify new platelet GPCRs, a platelet gene expression profile was generated and validated using quantitative real-time PCR. RESULTS: In total, mRNA of 28 GPCR genes was detected in human platelets. The 12 most abundant platelet GPCR transcripts were: thrombin receptor PAR1 (1865+/-178%), ADP receptor P2Y(12) (459+/-88%), succinate receptor 1 (257+/ 48%), ADP receptor P2Y(1) (100%), orphan P2RY(10) (68.2+/-3.3%), lysophosphatidic acid receptors GPR23 (48.2+/-11%) and GPR92 (26.1+/-3.3%), adrenergic receptor alpha(2A) (18.4+/-4.4%), orphan EBI2 (6.31+/-0.42), adenosine receptors A(2A) (2.94+/-0.24%) and A(2B) (2.88+/-0.16%) and lysophosphatidic acid receptor LPA(1) (2.59+/-0.39%) (% relative to the chosen calibrator P2Y(1)). A surprising G protein coupled receptor redundancy was found: two ADP receptors (P2Y(1) and P2Y(12)), three adenosine receptors (A(2A), A(2B), and A(1)), four lysophosphatidic acid receptors (LPA(1), LPA(3), GPR23 and GPR92), two l glutamate receptors (mGlu(3) and mGlu(4)) and two serotonin receptors (5-HT(1F) and 5-HT(4)). The adenosine receptor A(2B) gene expression was validated with protein expression and functional studies. Western blot confirmed A(2B) receptor protein expression and platelet flow cytometry demonstrated inhibition of the effect of NECA by the adenosine A(2B)-antagonist MRS1754. CONCLUSIONS: We have detected several GPCRs not previously known to be expressed in platelets, including a functional adenosine A(2B) receptor. The findings could improve our understanding of platelet aggregation and provide new targets for drug development. PMID- 17920664 TI - A natural histone deacetylase inhibitor, Psammaplin A, induces cell cycle arrest and apoptosis in human endometrial cancer cells. AB - OBJECTIVE: Histone deacetylase (HDAC) inhibitors are promising new class of anticancer agents that act by inhibiting cell proliferation and inducing cell cycle arrest of various cancer cells. Psammaplin A (PsA) is a phenolic natural product that has been isolated from marine sponges, and has been suggested to be a promising novel HDAC inhibitor. However, the precise mechanism of PsA as a HDAC inhibitor is poorly understood. This study investigated the anti-tumor effect of PsA on endometrial human cancer cells. METHODS: The cell proliferation, cell cycle, and apoptosis were measured in Ishikawa endometrial cancer cells after PsA treatment. RESULTS: PsA significantly inhibited the proliferation of Ishikawa cells in a dose-dependent manner. PsA markedly induced the expression of acetylated H3 and H4 histone proteins. In addition, PsA markedly up-regulated the expression of cyclin-dependent kinase inhibitor, p21(WAF1), and down-regulated the expression of pRb, cyclins, and CDKs, which lead to induce cell cycle arrest. Cell cycle analysis indicated that PsA treatment increased the proportion of cells in the G0/G1 and G2/M phases, and decreased the ratio of cells in the S phase. CONCLUSION: The PsA treatment resulted in the significant induction of apoptosis, which was associated with p53 independent p21(WAF1) expression. These results suggest that PsA exhibits the antiproliferative effects on endometrial cancer cells through selective induction of genes related to cell cycle arrest and apoptosis. PMID- 17920663 TI - Correlation between adiponectin and reduction of cell adhesion molecules after pitavastatin treatment in hyperlipidemic patients with type 2 diabetes mellitus. AB - The aim of this study was to determine whether pitavastatin may prevent the progression of atherosclerotic changes in hyperlipidemic patients. Seventy-five hyperlipidemic patients with and without type 2 diabetes were enrolled to receive pitavastatin 2 mg daily. Cell adhesion molecules (sCD40L, sP-selectin, sE selectin, and sL-selectin), chemokines (MCP-1 and RANTES) and adiponectin were measured at baseline and after 3 and 6 months of pitavastatin treatment. Adiponectin levels prior to pitavastatin treatment in hyperlipidemic patients with and without diabetes were lower than levels in normolipidemic controls. Both total cholesterol and the LDL-cholesterol (LDL-C) decreased significantly after pitavastatin administration. Additionally, hyperlipidemic patients with type 2 diabetes exhibited a significant increase in adiponectin levels after pitavastatin treatment (before vs. 3 months, 6 months, 2.81+/-0.95 vs. 3.84+/ 0.84 microg/ml (p<0.01), 4.61+/-1.15 mug/ml (p<0.001)). Furthermore, hyperlipidemic diabetics exhibited significant decreases in sE-selectin and sL selectin levels after 6 months of pitavastatin treatment (sE-selectin, before vs. 6 months, 74+/-21 vs. 51+/-10 ng/ml, p<0.05; sL-selectin, before vs. 6 months, 896+/-141 vs. 814+/-129 ng/ml, p<0.05). In addition, adiponectin showed significant correlation with sE-selectin and sL-selectin in diabetic hyperlipidemia. However, MCP-1, RANTES and sCD40L did not exhibit any differences before or after pitavastatin administration. These results suggest that pitavastatin possesses an adiponectin-dependent anti-atherosclerotic effect in hyperlipidemic patients with type 2 diabetes in addition to its lowering effects on total cholesterol and LDL-C. PMID- 17920665 TI - 5-HT6 receptor antagonist reversal of emotional learning and prepulse inhibition deficits induced by apomorphine or scopolamine. AB - 5-HT6 receptors have been implicated in consolidation of visuospatial and reward based learning tasks. Since 5-HT6 receptors may be important in modulation of sensory gating which is often affected in schizophrenic patients, we tested whether Ro 4368554, a 5-HT6 selective antagonist at a dose of 10 mg/kg, could reverse the loss of prepulse inhibition from apomorphine or scopolamine. In addition, we also tested whether Ro 4368554 altered fear conditioning using fear potentiated startle, a model for emotional learning. Prepulse inhibition of startle was disrupted by apomorphine (0.5 mg/kg) when prepulse emissions were 5 dB above background but not above 15 dB, while scopolamine (0.5 mg/kg) caused disruption at both prepulse levels. Scopolamine-mediated disruption was not reversed by Ro 4368854 but apomorphine-mediated disruption was significantly ameliorated by 5-HT6 inhibition. For fear potentiated startle, scopolamine and/or Ro 4368554 were administered before two daily fear conditioning sessions; rats were tested on the following day. Rats that received scopolamine displayed no fear potentiated startle but Ro 4368554 reversed this scopolamine deficit. Additionally, we mapped Fos induction in rats treated with scopolamine and/or Ro 4368554; scopolamine increased Fos expression in the central nucleus of the amygdala and this was attenuated by Ro 4368554. In summary, we have demonstrated the efficacy of 5-HT6 antagonists in modulating sensory gating and fear conditioning, and thus may be of therapeutic use for schizophrenia-related disorders. PMID- 17920666 TI - Regulatory dendritic cells protect against allergic airway inflammation in a murine asthmatic model. AB - BACKGROUND: Dendritic cells (DCs) are crucial for the induction of immunity and tolerance. Despite an improved understanding of the DC-mediated control of T(H)1 biased immunity, little is known about how DCs regulate T(H)2-mediated immunity. OBJECTIVE: The effects of immunostimulatory mature DCs (maDCs) and regulatory DCs (DCregs) on T(H)2-driven allergic immunity involving IgE production were examined. METHODS: A murine model of airway hyperresponsiveness; the adoptive transfer of maDCs, DCregs, and T cells; and T-cell function were studied. RESULTS: Antigen-pulsed maDCs inhibited antigen-specific IgE production but enhanced the production of antigen-specific IgG1 and IgG2a. Analysis of Ifng-/- mice and Il21r-/- mice revealed that the inhibitory effect of antigen-pulsed maDCs on antigen-specific IgE production involved IL-21-producing T follicular helper cells but not IFN-gamma-producing T(H)1 cells. In contrast, antigen-pulsed DCregs impaired the production of antigen-specific IgE, IgG1, and IgG2a. In vivo blockade experiments showed that antigen-specific CD4+CD25+Foxp3+ regulatory T cells mainly mediated the suppressive effect of antigen-pulsed DCregs on the production of antigen-specific IgE. Antigen-pulsed maDCs promoted airway inflammation, whereas antigen-pulsed DCregs markedly suppressed the pathogenesis. CONCLUSION: DCregs abolish T(H)2-mediated IgE production and allergic inflammation based on antigen-specific dominant tolerance, whereas maDCs exacerbate the pathogenesis despite inhibiting the IgE response through the activation of diverse types of T(H) cell responses. PMID- 17920668 TI - Childhood Asthma Control Test in asthmatic children with dysfunctional breathing. PMID- 17920669 TI - CD4/CD8 T-cell ratio predicts HIV infection in infants: the National Heart, Lung, and Blood Institute P2C2 Study. AB - BACKGROUND: In resource-poor regions of the world, HIV virologic testing is not available. OBJECTIVE: We sought to evaluate the diagnostic usefulness of the CD4/CD8 T-cell ratio in predicting HIV infection in infants. METHODS: Data from the 3- and 9-month visits for non-breast-fed infants born to HIV-infected mothers enrolled (1990-1994) in the Pediatric Pulmonary and Cardiac Complications of Vertically Transmitted HIV Infection Study (mother-to-child transmission of HIV, 17%) were analyzed. Data from the 3-month visit for infants enrolled (1985-1996) in the Perinatal AIDS Collaborative Transmission Study (mother-to-child transmission of HIV, 18%) were used for validation. RESULTS: At 3 months of age, data were available on 79 HIV-infected and 409 uninfected non-breast-fed infants in the Pediatric Pulmonary and Cardiac Complications of Vertically Transmitted HIV Infection Study. The area under the curve (AUC) of the receiver operating characteristic curve at 3 months was higher for the CD4/CD8 ratio compared with the CD4(+) T-cell count (AUC, 0.83 and 0.75; P = .03). The mean CD4/CD8 ratio at the 3-month visit was 1.7 for HIV-infected infants and 3.0 for uninfected infants. A CD4/CD8 ratio of 2.4 at 3 months of age was almost 2.5 times more likely to occur in an HIV-infected infant compared with an uninfected infant (test sensitivity, 81%; posttest probability of HIV, 33%). Model performance in the Centers for Disease Control and Prevention Perinatal AIDS Collaborative Transmission Study validation test (224 HIV-infected and 1015 uninfected 3-month old infants) was equally good (AUC, 0.78 for CD4/CD8 ratio). CONCLUSION: The CD4/CD8 T-cell ratio is a more sensitive predictor of HIV infection in infants than the CD4(+) T-cell count. CLINICAL IMPLICATIONS: The CD4/CD8 T-cell ratio can be used with caution to predict HIV infection in children. PMID- 17920670 TI - The effect of corticosteroids on the disposal of long-acting beta2-agonists by airway smooth muscle cells. AB - BACKGROUND: Organic cation transporters (OCTs) have an important role in tissue distribution and elimination of cationic drugs. Carrier-mediated disposal of cationic bronchodilators in the airway tissue, however, is incompletely understood. OBJECTIVES: We sought to assess the uptake of long-acting beta(2) agonist bronchodilators by bronchial and vascular smooth muscle cells. METHODS: Human airway cells and tissues obtained from organ donors were evaluated for cationic drug transporter expression by means of quantitative RT-PCR and immunofluorescence. For in vitro functional studies, tritiated formoterol and tritiated salmeterol uptake by bronchial and vascular smooth muscle cells was measured. RESULTS: Quantitative RT-PCR analysis revealed high mRNA levels for the corticosteroid-sensitive OCT3 in bronchial and vascular smooth muscle cells. Immunofluorescence staining of airway sections confirmed OCT3 expression in these cells. In bronchial smooth muscle cells, uptake of the cationic formoterol was inhibited with OCT inhibitors. Corticosteroids also inhibited formoterol uptake through a rapid (within 15 minutes) nongenomic action, with the following rank order for inhibitory potency: corticosterone > budesonide > fluticasone. The corticosteroid-induced inhibition was significantly higher in vascular than bronchial smooth muscle cells. In comparison with formoterol, uptake of the noncharged lipophilic salmeterol was approximately 10-fold higher and insensitive to all OCT inhibitors and corticosteroids. CONCLUSIONS: Our findings suggest that corticosteroids, through OCT3 inhibition, rapidly interfere with the disposal of cationic drugs by smooth muscle cells in the airway. CLINICAL IMPLICATIONS: This novel immediate interaction between corticosteroids and cationic beta(2)-agonist bronchodilators supports the use of such combinations in the pharmacotherapy of asthma. PMID- 17920671 TI - Accessibility and connectivity in physical activity studies: the impact of missing pedestrian data. AB - OBJECTIVE: One important characteristic in physical activity research into the built environment is network connectivity, usually calculated using street networks. However, a true pedestrian network may have very different connectivity than a street network. This study, conducted in 2004, examines the difference in walkability analyses when street networks versus pedestrian networks are used for four metropolitan suburbs in Perth, Western Australia. METHODS: A street network of Perth was used to represent the current standard of data for walkability analyses. Aerial photography from 2003 was used to create a pedestrian network, which incorporated pedestrian footpaths into the street network. The street and pedestrian networks were compared using three measures of connectivity: Pedsheds, link node ratio and pedestrian route directness. RESULTS: A comparison of the results using street versus pedestrian networks showed very different outcomes for conventional neighbourhood designs. Connectivity measures for conventional neighbourhoods improved up to 120% with the addition of pedestrian networks, although traditional neighbourhoods still had slightly better connectivity values overall. CONCLUSION: The true pedestrian network increases the connectivity of a neighbourhood and may have significant impact on these measures, especially in neighbourhoods with conventional street designs. It is critical that future studies incorporate pedestrian networks into their analyses. PMID- 17920672 TI - Selection of developmentally competent buffalo oocytes by brilliant cresyl blue staining before IVM. AB - The brilliant cresyl blue (BCB) test determines the activity of glucose-6 phosphate dehydrogenase (G6PDH); the activity of this enzyme is greatest in growing oocytes, but it declines as oocytes mature. The objective was to develop and evaluate this test for assessing development of buffalo oocytes (to select developmentally competent oocytes for increased in vitro embryo production). Oocytes were exposed to BCB stain diluted in mDPBS (DPBS with 0.4% BSA) for 90 min at 38.5 degrees C in a humidified air atmosphere; those with or without blue coloration of the cytoplasm were designated as BCB+ and BCB-, respectively. In Experiment 1, oocytes were exposed to 13, 26, or 39 microM BCB. There were fewer BCB+ oocytes after exposure to 13 microM BCB (10%) than after exposure to 26 or 39 microM BCB (57.2 and 61.8%; P<0.05), but there was no significant difference among treatments for blastocyst production rate. In Experiment 2, the diameter of BCB+ oocytes (144.4+/-4.2 microm; mean+/-S.E.M.) was higher (P<0.05) than that of BCB- oocytes (136.8+/-4.6 microm). In Experiment 3, oocytes were allocated into three groups: control (immediately cultured); holding-control (kept in mDPBS for 90 min before cultured); and treatment-incubation (incubated with 26 microM BCB). After IVM, oocytes were fertilized in vitro and cultured on an oviductal monolayer. The nuclear maturation rate was higher (P<0.05) in BCB+ (86.2%), control (83.4%) and holding-control (82.6%) oocytes than BCB- (59.2%) oocytes. The BCB+ oocytes yielded more blastocysts than control or holding-control oocytes (33.4, 20.2, and 21.0%, P<0.05); blastocyst development was lowest in BCB- oocytes (5.2%). In conclusion, staining of buffalo oocytes with BCB before IVM may be used to select developmentally competent oocytes for increased in vitro embryo production. PMID- 17920674 TI - Polysaccharides from the green seaweeds Codium fragile and C. vermilara with controversial effects on hemostasis. AB - Codium fragile and Codium vermilara biosynthesize water-soluble sulfated arabinans and galactans (and/or sulfated arabinogalactans), alpha(1-->4)-D glucans and beta(1-->4)-D-mannans. The former polysaccharides are composed by 3 linked beta-D-galactopyranose and beta-L-arabinopyranose residues, they are highly sulfated and substituted with pyruvic acid ketals. For both seaweeds, they have the same main structural units, but in different percentages. All the room temperature water extracts from both seaweeds showed a dual haemostatic effect: they prevented coagulation, but they induced platelet aggregation. Anticoagulant activity and platelet aggregation were higher in the samples with polysaccharides richer in sulfate, mainly in those from C. vermilara, which have a higher degree of sulfation and arabinose content. PMID- 17920673 TI - A polymerase chain reaction for detection of Brucella canis in vaginal swabs of naturally infected bitches. AB - A PCR assay for the detection of Brucella canis in canine vaginal swab samples was evaluated, comparing its performance with that of bacterial isolation, serological tests, and a blood PCR assay. One hundred and forty-four female dogs were clinically examined to detect reproductive problems and they were tested by the rapid slide agglutination test, with and without 2-mercaptoethanol (2ME-RSAT and RSAT, respectively). In addition, microbiological culture and PCR were performed on blood and vaginal swab samples. The results of the vaginal swab PCR were compared to those of the other tests using the Kappa coefficient and McNemar test. Of the 144 females that were examined, 66 (45.8%) were RSAT positive, 23 (15.9%) were 2ME-RSAT positive, 49 (34.02%) were blood culture positive, 6 (4.1%) were vaginal swab culture positive, 54 (37.5%) were blood PCR positive, 52 (36.2%) were vaginal swab PCR positive, and 50.69% (73/144) were positive by the combined PCR. The PCR was able to detect as few as 3.8 fg of B. canis DNA experimentally diluted in 54 ng of canine DNA, extracted from vaginal swab samples of non-infected bitches. In addition, the PCR assay amplified B. canis genetic sequences from vaginal swab samples containing 1.0 x 10(0) cfu/mL. In conclusion, vaginal swab PCR was a good candidate as a confirmatory test for brucellosis diagnosis in bitches suspected to be infected, especially those negative on blood culture or blood PCR; these animals may be important reservoirs of infection and could complicate attempts to eradicate the disease in confined populations. PMID- 17920675 TI - The structure and functionality of contractile forisome protein aggregates. AB - Although they have been discovered decades ago, only in the last years forisome protein aggregates received broader attention due to their ability to convert chemical into mechanical energy. In contrast to most other motor proteins, these proteins from Fabaceae plants are independent of high-energy chemical compounds, like e.g. ATP, but undergo an anisotropic shape transition (longitudinally expanded to contracted) in response to ion concentration changes (Ca(2+), H(+), etc.), instead. We present morphological and functional data on forisomes obtained using atomic force microscopy (AFM). High-aspect ratio AFM tips allow the detailed elucidation of structural characteristics that are inaccessible with standard AFM tips. Microindentation measurements were employed to calculate the elasticity of the forisome material. Young's moduli were found to be approximately 32.7 kPa in the expanded state and approximately 2.748 kPa in the contracted state of the polymer. These results are compared to investigations where a tipless AFM cantilever was utilized to exert a load against the shape transition. In the latter experiments, an energy conversion of approximately 2.29 pJ per stroke was detected. Energetical considerations support the hypothesis that the switching process is accompanied by a change in cross-linking of the constituent subunits and allow estimating the extent of cooperativity during the pH-induced transition. Finally, useful parameters were identified and characterized that are crucial for the application of forisomes as functional elements in microfluidic chips. PMID- 17920676 TI - Human mesenchymal stem cell differentiation to NP-like cells in chitosan glycerophosphate hydrogels. AB - Intervertebral disc (IVD) degeneration is one of the major causes of low back pain. As current clinical treatments are aimed at restoring biomechanical function and providing symptomatic relief, interest in methods focused on biological repair has increased. Several tissue engineering approaches using different cell types and hydrogels/scaffolds have been proposed. Owing to the unsuitable nature of degenerate cells for tissue engineering attention has focused on the use of mesenchymal stem cells (MSCs). Additionally, while rigid scaffolds have been demonstrated to allow MSC differentiation to the chondrocyte like cells of the IVD, hydrogels are being increasingly studied as they allow minimally invasive implantation without extensive damage to the IVD. Here, we have studied the temperature-sensitive hydrogel chitosan-glycerophosphate (C/Gp), seeded with human MSCs and cultured for 4 weeks in standard medium. We have analysed the gene and protein expression profile of the MSCs and compared it to that of both nucleus pulposus (NP) cells and articular chondrocytes cultured in C/Gp. Gene expression analysis for chondrocytic-cell marker genes demonstrated differentiation of MSCs to a phenotype which showed similarities to both articular chondrocytes and NP cells. Conventional PCR demonstrated a lack of expression of osteogenic marker genes and the hypertrophic marker gene type X collagen. MSCs also secreted both proteoglycans and collagens in a ratio, which more closely resembled that of NP cells than articular chondrocytes. These results therefore suggest that MSC-seeded C/Gp gels could be used clinically for the regeneration of the degenerate human IVD. PMID- 17920677 TI - TRPC channels determine human keratinocyte differentiation: new insight into basal cell carcinoma. AB - Aberrant keratinocyte differentiation is considered to be a key mechanism in the onset of hyperproliferative dermatological diseases, including basal cell carcinoma (BCC). It is, therefore, vital to understand what drives keratinocytes to develop such pathological phenotypes. The role of calcium in keratinocyte differentiation is uncontested but the mechanisms controlling calcium-induced differentiation have yet to be completely elucidated. This study was designed to investigate the role of calcium-permeable TRPC channels in human keratinocyte differentiation and BCC, using a combination of molecular and cell biology approaches, involving electrophysiology and Ca(2+)-imaging, on the HaCaT cell line, primary cultures of normal human keratinocytes, and BCC cells. We demonstrated that TRPC1/TRPC4 channel expression was important for keratinocyte differentiation, as knocking out these channels (by siRNA strategy) prevented the induction of Ca(2+)-induced differentiation. TRPC1/TRPC4-mediated calcium entry and endoplasmic reticulum Ca(2+) content increased significantly in differentiated keratinocytes. However, the failure of BCC cells to differentiate was related to a lack of TRPC channel expression and calcium entry. In summary, our data demonstrate that TRPC1 and TRPC4 channels are key elements in keratinocyte Ca(2+) homeostasis and differentiation and may therefore be responsible for skin pathologies. PMID- 17920678 TI - Up-regulation of NKG2D ligands by AML cells to increase sensitivity to NK cells: the tumour might strike back. Comment on "differentiation-promoting drugs up regulate NKG2D ligand expression and enhance the susceptibility of acute myeloid leukemia cells to natural killer cell-mediated lysis" by Rohner et al. [Leuk Res 2007;31:1393-402]. PMID- 17920679 TI - Arsenic trioxide, thalidomide and retinoid acid combination therapy in higher risk myelodysplastic syndrome patients. AB - OBJECTIVE: To evaluate the clinical efficacy and safety of arsenic trioxide, retinoic acid and thalidomide combination therapy in higher risk MDS. METHODS: Twenty-one patients diagnosed with higher risk MDS were administered 10mg/day arsenic trioxide intravenously for 10 days, 40mg/day retinoic acid orally for 2 weeks and 100mg/day thalidomide orally for 4 weeks per cycle. RESULTS: After at least two treatment cycles, 10 patients showed hematologic responses. One achieved CR, one achieved PR, three patients achieved major hematological improvements. The efficacy rate was 24% (5/21), and the response rate was 48% (10/21). The schedule was tolerated well by all patients and toxicities were moderate and reversible. CONCLUSION: The combination of arsenic trioxide, retinoic acid and thalidomide could have therapeutic benefit in higher risk MDS with safety. PMID- 17920680 TI - Low dose rapamycin does not modulate p-glycoprotein function in acute myeloid leukaemia. PMID- 17920681 TI - Antioxidative role of selenium on some tissues of (Cd2+), Cr3+)-induced rainbow trout. AB - Chemical toxic pollutants (especially heavy metals) are important sources of reactive oxygen species (ROS) in biological systems. Membrane phospholipids of aerobic organisms are continually subjected to oxidant challenges from endogenous and exogenous sources, while peroxidized membranes and lipid peroxidation products represent constant threats to aerobic cells. The primary antioxidant protection against free radical and ROS is provided by the enzymes glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT), respectively. The trace element selenium has been implicated in chemo-prevention and drug resistance through reduction of oxidative stress. Selenium could prevent damage to the unsaturated fatty acid of subcellular membranes by lipid peroxidation induced by free radicals. The results reported here show that sodium selenite has an important contribution to antioxidative defense for the spleen and heart of rainbow trout. The ability of sodium selenite to prevent the oxidative stress induced by heavy metals (Cd(2+), Cr(3+)) in fish was rationalized. PMID- 17920682 TI - Evaluation of biochemical responses in Palearctic and Lake Baikal endemic amphipod species exposed to CdCl2. AB - This study evaluated small heat shock proteins (sHSP) (related to alpha crystallin) and antioxidant enzymes (POD, peroxidase and CAT, catalase) as possible biomarkers for use in toxicological studies. Biochemical responses to cadmium chloride in two Lake Baikal endemic amphipods (Eulimnogammarus verrucosus, Eulimnogammarus cyaneus) and Palearctic species (Gammarus lacustris) were compared. Our findings showed that cadmium chloride toxicity directly influenced POD activity and sHSP synthesis in all amphipod species. The Baikalean endemic and the Palearctic amphipod species responded by decreasing activity of POD and they exhibited a dose-dependent activation of sHSP synthesis. All measured parameters differed among species and depended on the species' ability to resist cadmium chloride toxicity. CAT activity in the Palearctic species responded significantly to cadmium chloride exposure; however, responses were negligible for both Baikalean species. We suggest that synthesis of sHSP, together with changes in POD activity, could be used as biomarkers for further studies of amphipod species including endemics from Lake Baikal. PMID- 17920683 TI - Identification of somatic hypermutations in the TP53 gene in B-cell chronic lymphocytic leukemia. AB - Abnormalities of the TP53 gene are associated with a particularly severe prognosis in patients with B-cell chronic lymphocytic leukemia (B-CLL). This tumor-suppressor is mostly inactivated by the deletion of one and point mutation of the other allele and has not been previously shown to be hypermutated in B CLL. We identified two patients whose lymphocytes showed repeatedly an extensive proportion of TP53 mutated cells by FASAY analysis (the yeast functional assay) and harbored various TP53 mutations, mostly single-base substitutions, in individual cells. The mutation targeting exhibited characteristic traits of the somatic hypermutation process. In the first patient (harboring the unmutated IgVH locus) a significant bias to point mutations at CG pairs (21/25; P=0.009), their remarkable preference for the RGYW/WRCY motives (28%) and the highest expression of the activation-induced cytidine deaminase (AID) mRNA among the 34 tested B-CLL samples. In the second patient no CG bias was observed but the targeting of point mutations into the RGYW/WRCY motives was even more prominent here (7/16; 44%). Moreover, six out of eight point mutations affecting AT pairs were localized in the WA/TW motives, which are also characteristic for the somatic hypermutations. This patient, who was IgVH-mutated, already did not express any significant amount of the AID transcript. Our findings add a new aspect to the mosaic of the p53 mutability in B-CLL. PMID- 17920684 TI - SOCS3 promotes TLR4 response in macrophages by feedback inhibiting TGF beta1/Smad3 signaling. AB - Endogenous transforming growth factor-beta1 (TGF-beta1) plays an important role in the negative regulation of toll-like receptor (TLR) signaling in a feedback manner. Suppressors of cytokine signaling 3 (SOCS3) has been shown to be induced by TGF-beta1 in osteoclast/macrophage, while the reports on the role of SOCS3 in regulating TLR4 signaling were controversial. The functional relationship between SOCS3 and TGF-beta1/Smad3 pathway in TLR4 response also remains unclear. In this study, we demonstrate that LPS-induced endogenous TGF-beta1 contributes to the inducible SOCS3 expression in macrophages. SOCS3 silencing could markedly decrease the LPS-induced production of TNF-alpha and IL-6 in macrophages. Interestingly, less decrease of LPS-induced TNF-alpha, IL-6 by SOCS3 silencing was observed in Smad3 null macrophages. Furthermore, we found SOCS3 could interact with Smad3, and inhibit Smad3 nuclear translocation and transcriptional activity. Therefore, our data demonstrate that SOCS3 is a positive regulator of TLR4 response by feedback inhibiting endogenous TGF-beta1/Smad3 signaling, thus outlining a new feedback regulatory manner for TLR4 response in macrophages. PMID- 17920685 TI - DNA interaction of some polymer-copper(II) complexes containing 2,2'-bipyridyl ligand and their antimicrobial activities. AB - The water soluble polymer-copper(II) complex samples, [Cu(bpy)(2)(BPEI)]Cl(2).4H(2)O (bpy=2,2'-bipyridine, BPEI=branched polyethyleneimine), with varying degrees of copper(II) chelates content in the polymer chain, were prepared by ligand substitution method in water-ethanol medium and characterized by Infra-red, UV-visible, EPR spectral and elemental analysis methods. The interaction of these polymer-copper(II)-bipyridyl complex samples with calf thymus DNA has been explored by using electronic absorption spectroscopy, emission spectroscopy and gel electrophoresis techniques. The observed changes in the physico-chemical features of the polymer-copper(II) complex on binding to DNA suggest that the complex binds to DNA with electrostatic interaction mode. A sample of polymer-copper(II) complex was tested for its antibacterial and antifungal activity and it was found to have good antibacterial and antifungal activities. PMID- 17920686 TI - Mortality associated with central nervous system tuberculosis. AB - BACKGROUND: Tuberculosis (TB) of the central nervous system (CNSTB) is associated with higher mortality rates than other forms of TB. Epidemiologic associations with and prognostic indicators of CNSTB have not been assessed in a large US population-based study. METHODS: Between 1995 and 2004 and using a population based active surveillance study, we compared patients with CNSTB to patients with TB affecting sites other than CNS (non-CNSTB) with respect to sociodemographic, clinical and Mycobacterium tuberculosis genotype variables. Risk factors associated with mortality at 180 days were compared between the 2 groups. RESULTS: We enrolled 92 patients with CNSTB and 3570 with non-CNSTB. HIV co infection was present in 31 (33.7%) of the CNSTB cases. In a Cox proportion hazard model, we found that CNSTB patients who died within 180 days were more likely to be older (HR 1.06, 95% CI 1.02-1.10), have a positive MTB culture from a CNS source (HR 5.11, 95% CI 1.06-24.62) and have hydrocephalus (HR 10.62, 95% CI 3.28-34.36) than patients who survived CNSTB. HIV co-infection association with mortality was not statistically significant (HR 1.74, 95% CI 0.35-8.62). CONCLUSIONS: In our cohort, hydrocephalus was the most important predictor of mortality post-CNSTB diagnosis. PMID- 17920687 TI - Mycobacterium other than tuberculosis (MOTT) infection: an emerging disease in infliximab-treated patients. AB - OBJECTIVES: Infliximab has revolutionized treatment of rheumatologic diseases and inflammatory bowel disease. However, it increases the risk of tuberculosis. Less is known about the development of Mycobacterium other than tuberculosis (MOTT) infection. We review the literature on non-tuberculous mycobacterial infections in infliximab-treated patients and report the first case of disseminated Mycobacterium avium complex in an infliximab-treated patient complicated by immune reconstitution inflammatory syndrome. METHODS AND RESULTS: MEDLINE search with the keywords mycobacteria and infliximab revealed four cases of MOTT in patients treated with infliximab: fatal Mycobacterium peregrinum pneumonia in a patient with polymyositis and dermatomyositis; a patient with rheumatoid arthritis with skin and soft tissue infection with Mycobacterium abscessus; Mycobacterium fortuitum in a patient with rheumatoid arthritis; and a case of pulmonary MAC without dissemination. Review of US data from 1998 to 2002 published by Wallis et al. revealed that out of more than 233,000 patients treated with infliximab, 30 developed unspecified mycobacterial species infection. No further data was available regarding these cases. CONCLUSION: MOTT infection is a rare but emerging complication of infliximab therapy. MOTT cases tend to progress rapidly in infliximab-treated patients and withdrawal of infliximab therapy can result in immune reconstitution. PMID- 17920688 TI - Changes in neuroactive steroid content during social isolation stress modulate GABAA receptor plasticity and function. AB - Rats deprived of social contact with other rats at a young age experience a form of prolonged stress that leads to long-lasting alteration in their behavior profile. This chronic stress paradigm is thus thought to be anxiogenic for these normally gregarious animals and their abnormal reactivity to environmental stimuli, when reared under this condition, is thought to be a product of prolonged stress. Neurochemical, molecular, and electrophysiological evidences demonstrate that social isolation is associated with alteration in the structure and function of GABA(A) receptors and suggest that endogenous content of the progesterone metabolite 3alpha,5alpha-TH PROG may be an important determinant in regulating brain excitability and sensitivity to stimuli and point out its possible role in psychiatric and neurological disorder. PMID- 17920689 TI - Organisation of the spinal central pattern generators for locomotion in the salamander: biology and modelling. AB - Among living tetrapods, salamanders are regarded as most closely resembling the first terrestrial vertebrates, and are therefore an interesting group in which the evolutionary changes in the locomotor behaviour from aquatic to terrestrial habitats can be inferred. Salamanders exhibit two locomotor modes: swimming and terrestrial stepping. The swimming is anguilliform and resembles closely that of the lamprey. On the ground, the salamander switches to a stepping gait with axial undulations that is also observed in many reptiles. The salamander is therefore ideally suited for examining the neural mechanisms for the generation of these two locomotor modes, as well as the neural mechanisms of gait transition. In the present paper, we describe the kinematics and patterns of activation of axial and limb muscles during stepping and swimming in adult salamanders. We then review the current neurobiological data about the organisation of the spinal networks underlying swimming and stepping, and the mechanisms of gait transition. Finally we report modelling studies aimed at understanding the organisation and operation of the salamander locomotor circuits. Altogether, the neurobiological and the modelling data support the hypothesis of a phylogenetic conservatism from agnathians to amphibians of the spinal locomotor networks generating axial motor patterns. PMID- 17920690 TI - Common pathological processes in Alzheimer disease and type 2 diabetes: a review. AB - Alzheimer disease (AD) and type 2 diabetes mellitus (T2DM) are conditions that affect a large number of people in the industrialized countries. Both conditions are on the increase, and finding novel treatments to cure or prevent them are a major aim in research. Somewhat surprisingly, AD and T2DM share several molecular processes that underlie the respective degenerative developments. This review describes and discusses several of these shared biochemical and physiological pathways. Disturbances in insulin signalling appears to be the main common impairment that affects cell growth and differentiation, cellular repair mechanisms, energy metabolism, and glucose utilization. Insulin not only regulates blood sugar levels but also acts as a growth factor on all cells including neurons in the CNS. Impairment of insulin signalling therefore not only affects blood glucose levels but also causes numerous degenerative processes. Other growth factor signalling systems such as insulin growth factors (IGFs) and transforming growth factors (TGFs) also are affected in both conditions. Also, the misfolding of proteins plays an important role in both diseases, as does the aggregation of amyloid peptides and of hyperphosphorylated proteins. Furthermore, more general physiological processes such as angiopathic and cytotoxic developments, the induction of apoptosis, or of non-apoptotic cell death via production of free radicals greatly influence the progression of AD and T2DM. The increase of detailed knowledge of these common physiological processes open up the opportunities for treatments that can prevent or reduce the onset of AD as well as T2DM. PMID- 17920691 TI - High-performance liquid chromatographic determination of unreacted monomers and other residues contained in dental composites. AB - HPLC method was developed for determination of bisphenol A diglycidyl methacrylate (bis-GMA), bisphenol A diglycidyl acrylate (bis-GA), bisphenol A dimethacrylate (bis-DMA), glycidylmethacrylate (GMA) and triethylenglycol dimethacrylate (TEGDMA). Separation was carried out on a reversed phase Omnisphere 5 C18 column with a gradient mobile phase of CH3CN/H2O. UV detection was set at 205 nm and 275 nm parallel. The limits of quantification were found. The method has been applied for quantification of unreacted monomers trapped in polymer network of fillings. PMID- 17920692 TI - Predicting recovery from episodes of major depression. AB - BACKGROUND: This study examined psychosocial functioning as a predictor of recovery from episodes of unipolar major depression. METHODS: 231 subjects diagnosed with major depressive disorder according to Research Diagnostic Criteria were prospectively followed for up to 20 years as part of the NIMH Collaborative Depression Study. The association between psychosocial functioning and recovery from episodes of unipolar major depression was analyzed with a mixed effects logistic regression model which controlled for cumulative morbidity, defined as the amount of time ill with major depression during prospective follow up. Recovery was defined as at least eight consecutive weeks with either no symptoms of major depression, or only one or two symptoms at a mild level of severity. RESULTS: In the mixed-effects model, a one standard deviation increase in psychosocial impairment was significantly associated with a 22% decrease in the likelihood of subsequent recovery from an episode of major depression (OR=0.78, 95% CI: 0.74-0.82, Z=-3.17, p<0.002). Also, a one standard deviation increase in cumulative morbidity was significantly associated with a 61% decrease in the probability of recovery (OR=0.3899, 95% CI: 0.3894-0.3903, Z=-7.21, p<0.001). LIMITATIONS: The generalizability of the study is limited in so far as subjects were recruited as they sought treatment at academic medical centers. The analyses examined the relationship between psychosocial functioning and recovery from major depression, and did not include episodes of minor depression. Furthermore, this was an observational study and the investigators did not control treatment. CONCLUSIONS: Assessment of psychosocial impairment may help identify patients less likely to recover from an episode of major depression. PMID- 17920693 TI - Whole blood BDNF levels in healthy twins discordant for affective disorder: association to life events and neuroticism. AB - BACKGROUND: Depression has been associated with decreased blood BDNF concentrations; but it is unclear if low blood BDNF levels are a state or a trait marker of depression. METHODS: We investigated blood BDNF concentrations in a twin population including both subjects highly predisposed and protected against affective disorder. Whole blood assessed for BDNF concentrations and correlated to risk status, neuroticism, and number of stressful life events. RESULTS: Between the groups, we found no significant difference in whole blood BDNF levels. Women at high-risk for depression who had experienced three or more recent stressful events (n=26) had decreased whole blood BDNF levels compared to high-risk women with two or less recent stressful events (n=35), 21.6+/-7.0 vs. 18.5+/-4.1 ng/ml, respectively, (p<0.05). No such association was found in low risk women or in men. In men, however, low neuroticism scores and two or less recent stressful events were associated with decreased whole blood BDNF levels (n=50, p<0.05). LIMITATIONS: The cross-sectional design limits the possibility of drawing firm conclusions on causatility of the findings. CONCLUSION: The genetic risk of developing depression does not translate directly into whole blood BDNF levels. In females who are genetically disposed for depression and subjected to recent stressful life events whole blood BDNF levels are lower. PMID- 17920694 TI - Antibody treatment of human tumor xenografts elicits active anti-tumor immunity in nude mice. AB - Athymic nude mice bearing subcutaneous tumor xenografts of the human anti colorectal cancer cell line SW480 were used as a preclinical model to explore anti-tumor immunotherapies. Intratumor or systemic treatment of the mice with murine anti-SW480 serum, recombinant anti-SW480 polyclonal antibodies, or the anti-colorectal cancer monoclonal antibody CO17-1A, caused retardation or regression of SW480 tumor xenografts. Interestingly, when mice that had regressed their tumors were re-challenged with SW480 cells, these mice regressed the new tumors without further antibody treatment. Adoptive transfer of spleen cells from mice that had regressed their tumors conferred anti-tumor immunity to naive nude mice. Pilot experiments suggest that the transferred anti-tumor immunity is mediated by T cells of both gammadelta and alphabeta lineages. These results demonstrate that passive anti-tumor immunotherapy can elicit active immunity and support a role for extra-thymic gammadelta and alphabeta T cells in tumor rejection. Implications for potential immunotherapies include injection of tumor nodules in cancer patients with anti-tumor antibodies to induce anti-tumor T cell immunity. PMID- 17920695 TI - Effect of endogenous catecholamines on apoptosis of Con A-activated lymphocytes of rats. AB - Our previous studies show that lymphocytes express tyrosine hydroxylase (TH) and synthesize catecholamines (CAs) including dopamine, epinephrine and norepinephrine, and that the lymphocytes-derived endogenous CAs affect function of lymphocytes via autocrine/paracrine pathways. Over recent years, induction of apoptosis has been suggested to be a possible mechanism underlying the endogenous CAs-mediated lymphocyte proliferation, differentiation and activation. However, direct effect of the lymphocytes-synthesized CAs on lymphocyte apoptosis is less known. In the present study, TH inhibitor alpha-methyl-p-tyrosine (alpha-MT) and monoamine oxydase inhibitor pargyline were employed to block the synthesis and degradation of CAs in lymphocytes activated by concanavalin A (Con A). Apoptotic cells and apoptosis-related genes and proteins, Bax, Bcl-2, Fas, Fas-Ligand (FasL) and caspase-3, were examined in the lymphocytes treated with alpha-MT or pargyline by means of Annexin V/propidium iodide (PI) staining, real-time PCR and Western blot analyses, respectively. The treatment with alpha-MT of 10(-6) M and 10(-5) M (not 10(-7) M) notably reduced intracellular and supernatant DA, E and NE of the Con A-activated lymphocytes in a dose-dependent manner, and correspondingly, the treatment induced a remarkable decrease of apoptotic lymphocytes but not necrotic cells. The expression of Bax, Fas, FasL and caspase 3 mRNAs and proteins was significantly inhibited in the Con A-activated lymphocytes after the cells were treated with alpha-MT of 10(-6) M and 10(-5) M; but the expression of Bcl-2 mRNA and protein was dramatically increased by the alpha-MT treatment. Contrarily, the treatment with pargyline of 10(-6) M and 10( 5) M (not 10(-7) M) evidently increased the intracellular and supernatant DA, E and NE contents of the Con A-activated lymphocytes in a dose-dependent manner, and meanwhile, it caused a striking increase of apoptotic lymphocytes but not necrotic cells. The expression of Bax, Fas, FasL and caspase-3 mRNAs and proteins in the Con A-stimulated lymphocytes was remarkably enhanced by the treatment with pargyline of 10(-6) M and 10(-5) M, but the expression of Bcl-2 mRNA and protein was notably attenuated by the pargyline treatment. These results imply that endogenous CAs synthesized and secreted by lymphocytes accelerate lymphocyte apoptosis by altering fine balance between the expression of antiapoptotic and proapoptotic markers at transcriptional and translational levels, and suggest that both the death receptor pathway and the mitochondrial pathway are involved in the endogenous CAs-induced apoptosis. PMID- 17920696 TI - CD8+ T cells in inflammatory demyelinating disease. AB - We review the contribution made by CD8+ T cells to inflammation in the central nervous system (CNS) in Multiple Sclerosis (MS), and discuss their role in the animal model Experimental Autoimmune Encephalomyelitis (EAE). We show that the inflammatory cytokines interferon-gamma and interleukin-17 are differentially regulated in CNS-infiltrating CD4+ and CD8+ T cells in EAE, and that CD8+ T cells regulate disease. In MS, CD8+ T cells appear to play a role in promotion of disease, so cytokine regulation is likely different in CD8+ T cells in MS and EAE. PMID- 17920697 TI - Genetic analysis of the exon 1 position 49 CD152 dimorphism in multiple sclerosis. AB - Several studies have examined whether a dimorphism in the CD152 costimulatory molecule may influence the development of multiple sclerosis (MS). A sample of 108 patients with a diagnosis of relapsing remitting (RRMS), 28 with secondary progressive (SPMS), 23 with primary progressive (PPMS) and 63 people with no prior history of neurological conditions were selected from the MS clinic at the University of Texas Southwestern Medical Center at Dallas. Peripheral blood was separated with gradient extraction for leukocytes and genomic DNA extracted for CD152 A/G dimorphism analysis. A 163 bp PCR product in exon 1 including the position 49 A/G dimorphism was examined via single strand conformation polymophism (SSCP). Patient haplotype frequencies were compared between cases and controls and Pearson Chi-Square test performed to demonstrate statistical differences between MS groups and controls. Our results, similar to several recent studies, suggest that there is no statistical association with the risk of developing MS and no increased frequency in A or G at position 49 of exon 1 of CD152. Demonstration of prolonged proliferation in patient samples containing the GG genotypes and altered CD152 surface expression was also not demonstrated suggesting that the CD152 exon 1 position 49 A/G dimorphism does not contribute significantly to the development of MS in this patient population. PMID- 17920699 TI - Setting up hearing screening in meningitis children in Luanda, Angola. PMID- 17920700 TI - Hearing benefits of second-side cochlear implantation in two groups of children. AB - OBJECTIVE: To investigate the additional bilateral benefits of a second cochlear implant (CI) in a group of young children (<6 years of age) and a group of older children (>6 years of age). METHOD: This is a Belgian tertiary multi-centre study in which 33 CI-children with a second implant between the age of 2 and 12 participated. Assessments took place pre-second implant and at several time intervals post-fitting on pure tone audiometry and speech recognition in quiet and noise (+10 dBSNR). Testing was done with the first and second implant alone and bilaterally. Results were analysed separately for children younger and older than 6 years at the time of implantation of the 2nd CI. RESULTS: After 18 months of bilateral implant use all children obtained significantly higher hearing thresholds in the bilateral condition in comparison to both the unilateral conditions (p(CI1)=0.035/p(CI2)=0.042 for the younger children and p(CI1)=0.021/p(CI2)=0.007 for the older children). The speech recognition scores in quiet were for all children superior in the bilateral condition (p(CI2)=0.011 for the younger children and p(CI1)=0.016/p(CI2)=0.003 for the older children). In the noisy condition only significant bilateral better results were obtained in the group of younger children (p(CI1)=0.028/p(CI2)=0.034). CONCLUSIONS: Bilateral cochlear implantation offers advantages to all children. Even for the children who received a second implant after the age of 6 a progress is determined after 18 months. However, the data appear to show a beneficial performance for those children who received their second implant before the age of 6, especially in the more challenging conditions. PMID- 17920701 TI - A one-step reverse transcriptase loop-mediated isothermal amplification assay for simple and rapid detection of swine vesicular disease virus. AB - This report describes the development of a one-step reverse transcriptase loop mediated isothermal amplification (RT-LAMP) assay for the detection of swine vesicular disease virus (SVDV). The assay detects the virus rapidly, within 30-60 min and the result is visualised either by gel-electrophoresis or by the naked eye through the addition of SybrGreen. A collection of 28 SVDV isolates were tested positive, while heterologous viruses such as foot-and-mouth disease virus and vesicular stomatitis virus remained negative. The performance of the RT-LAMP was compared directly with real-time PCR using RNA from clinical samples including nasal swabs, serum and faeces. For nasal swabs and serum the sensitivity of the RT-LAMP was shown to be at least equivalent to real-time PCR. Interestingly, for faecal samples the RT-LAMP assay was shown to be even more sensitive than real-time PCR, possibly because it is less sensitive to inhibitory substances. This RT-LAMP assay provides a number of benefits for the diagnosis of SVD, since the assay is sensitive and rapid, and the isothermal amplification strategy used is not reliant upon expensive equipment it is particularly suited for "front line" diagnosis of SVD in modestly equipped laboratories, in field stations or in mobile diagnostic units. PMID- 17920698 TI - High cell surface expression of CD4 allows distinction of CD4(+)CD25(+) antigen specific effector T cells from CD4(+)CD25(+) regulatory T cells in murine experimental autoimmune encephalomyelitis. AB - Analysis of T regulatory cells (Treg) and T effector cells (Teff) in experimental autoimmune encephalomyelitis is complicated by the fact that both cell types express CD4 and CD25. We demonstrate that encephalitogenic T cells, following antigen recognition, up-regulate cell surface expression of CD4. The CD4(high) sub-population contains all of the antigen response as shown by proliferation and cytokine secretion, and only these cells are capable of transferring EAE to naive animals. On the other hand, a FACS separable CD25(+) sub-population of cells displayed consistent levels of CD4 prior to and after antigen stimulation. These cells displayed characteristics of Treg, such as expressing high levels of the Foxp3 gene and the ability to suppress mitogenic T cell responses. PMID- 17920702 TI - Quantitative PCR technique for detecting lymphocytic choriomeningitis virus in vivo. AB - Quantitative PCR (QPCR, or real time PCR (rtPCR)) has emerged as a powerful virologic technique for measuring viral replication and viral loads in humans and animal models. We have developed a QPCR assay to accurately quantify lymphocytic choriomeningitis virus (LCMV) in infected mice. We first validated this assay using plasmid DNA and LCMV viral stocks. We then demonstrated that the LCMV QPCR assay can detect LCMV in serum and tissues of chronically infected mice (LCMV clone 13), with greater sensitivity than conventional plaque assay. Subsequently, we demonstrated that the QPCR assay can detect LCMV in tissues of CD40L(-/-) mice during a low grade chronic infection with LCMV Armstrong. Finally, we improved the assay further such that it was approximate 1000-fold more sensitive than plaque assay for detection of the presence of LCMV in tissue. PMID- 17920703 TI - Oligonucleotide microarray-based detection and genotyping of Plum pox virus. AB - Plum pox virus (PPV) is the most damaging viral pathogen of stone fruits. The detection and identification of its strains are therefore of critical importance to plant quarantine and certification programs. Existing methods to screen strains of PPV suffer from significant limitations such as the simultaneous detection and genotyping of several strains of PPV in samples infected with different isolates of the virus. A genomic strategy for PPV screening based on the viral nucleotide sequence was developed to enable the detection and genotyping of the virus from infected plant tissue or biological samples. The basis of this approach is a long 70-mer oligonucleotide DNA microarray capable of simultaneously detecting and genotyping PPV strains. Several 70-mer oligonucleotide probes were specific for the detection and genotyping of individual PPV isolates to their strains. Other probes were specific for the detection and identification of two or three PPV strains. One probe (universal), derived from the genome highly conserved 3' non-translated region, detected all individual strains of PPV. This universal PPV probe, combined with probes specific for each known strain, could be used for new PPV strain discovery. Finally, indirect fluorescent labeling of cDNA with cyanine after cDNA synthesis enhanced the sensitivity of the virus detection without the use of the PCR amplification step. The PPV microarray detected and identified efficiently the PPV strains in PPV-infected peach, apricot and Nicotiana benthamiana leaves. This PPV detection method is versatile, and enables the simultaneous detection of plant pathogens. PMID- 17920704 TI - Lesions of the basal forebrain impair reversal learning but not shifting of attentional set in rats. AB - The cholinergic neurons of the basal forebrain, which project to cortex, the thalamic reticular nucleus and the amygdala, are implicated in many aspects of attentional function, while the intrinsic neurons of the basal forebrain are implicated in learning and memory. This study compared the effects of lesions of the basal forebrain made with either the immunotoxin 192-IgG-saporin (which selectively destroys cholinergic neurons), or the non-selective excitotoxin, ibotenic acid (which destroys both cholinergic and non-cholinergic neurons) on a task which measure the acquisition and shifting of attentional set as well as the ability to learn reversals of specific stimulus-reward pairings. Rats learned to obtain food reward by digging in small bowls containing distinctive digging media that were differentially scented with distinct odours. They performed a series of two-choice discriminations, with the bait associated with either the odour or the digging medium. Rats with 192-IgG-saporin lesions of the basal forebrain were not impaired relative to control rats at any stage of the task. Rats with ibotenic acid lesions of the basal forebrain were impaired the first time stimulus-reward contingencies were reversed. They were not impaired in acquisition of new discriminations, even when an attentional-shift was required. These data are consistent with data from marmosets and so highlight the functional similarity of monkey and rodent basal forebrain. They also confirm the likely involvement of non-cholinergic neurons of the basal forebrain in reversal learning. PMID- 17920705 TI - The role of PYY in feeding regulation. AB - Peptide YY (PYY), a 36-amino-acid peptide, is secreted primarily from L-cells residing in the intestinal mucosa of the ileum and large intestine. PYY, which belongs to a family of peptides including neuropeptide Y (NPY) and pancreatic polypeptide, is released into the circulation as PYY(1-36) and PYY(3-36); the latter is the major form of PYY in gut mucosal endocrine cells and throughout the circulation. Plasma PYY levels begin to rise within 15 min after starting to eat and plateau within approximately 90 min, remaining elevated for up to 6 h. Exogenous administration of PYY(3-36) reduces energy intake and body weight in both humans and animals. Via Y2 receptors, the satiety signal mediated by PYY inhibits NPY neurons and activates pro-opiomelanocortin neurons within the hypothalamic arcuate nucleus. Peripheral PYY(3-36) binds Y2 receptors on vagal afferent terminals to transmit the satiety signal to the brain. PYY(3-36) may have therapeutic potential in human obesity. PMID- 17920706 TI - Natriuretic peptides cause relaxation of human esophageal mucosal muscle. AB - Natriuretic peptides have been demonstrated to cause relaxation of the human gallbladder muscle through interaction with natriuretic peptide receptor-B (NPR B/NPR2). Effects of natriuretic peptides in the human esophageal muscle were unknown. To investigate the effects of natriuretic peptides in the human esophagus, we measured relaxation of muscularis mucosae strips isolated from the human esophagus caused by C-type natriuretic peptide (CNP), brain natriuretic peptide (BNP), atrial natriuretic peptide (ANP) and des[Gln(18), Ser(19), Gly(20), Leu(21), Gly(22)]ANP(4-23) amide (cANP(4-23)), a selective natriuretic peptide receptor-C (NPR-C) agonist. In endothelin-1 or carbachol-contracted mucosal muscle strips, CNP caused moderate, sustained and concentration-dependent relaxation. BNP caused a very mild relaxation whereas ANP and cANP(4-23) did not cause any relaxation. CNP was much more potent than BNP and ANP in causing relaxation. These suggest the existence of NPR-B mediating relaxation. The CNP induced relaxation was not affected by tetrodotoxin or atropine in endothelin-1 contracted esophageal strips and not by tetrodotoxin in carbachol-contracted strips, indicating a direct effect of CNP on the human esophageal muscularis mucosae. Taken together, these results demonstrate that natriuretic peptides cause relaxation of the muscularis mucosae of the human esophagus and suggest that the relaxation is through interaction with NPR-B. Natriuretic peptides may play an important role in the control of human esophageal motility. PMID- 17920707 TI - Effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on prolactin and somatolactin release from the goldfish pituitary in vitro. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) plays a role in mediating growth hormone and gonadotropin release in the teleost pituitary. In the present study, we examined the immunohistochemical relationship between PACAP nerve fibers and prolactin (PRL)- and somatolactin (SL)-producing cells in the goldfish pituitary. Nerve fibers with PACAP-like immunoreactivity (PACAP-LI) were identified in the neurohypophysis in close proximity to cells containing PRL-LI or SL-LI. Several cells with PRL-LI or SL-LI showed PACAP receptor (PAC(1)R)-LI. The cell immunoblot assay method was used to examine the effect of PACAP on PRL and SL release from dispersed goldfish pituitary cells. Treatment with PACAP increased the immunoblot area for PRL- and SL-LI from individual pituitary cells in a dose-dependent manner. The effect of PACAP on the expression of mRNAs for PRL and SL in cultured pituitary cells was also tested. Semiquantitative analysis revealed that the expression of SL mRNA, but not PRL mRNA, was increased significantly by the treatment with PACAP. The effect of PACAP on intracellular calcium mobilization in isolated pituitary cells was also investigated using confocal laser-scanning microscopy. The amplitude of Ca(2+) mobilization in individual cells showing PRL- or SL-LI was increased significantly following exposure of cells to PACAP. These results indicate that PACAP can potentially function as a hypophysiotropic factor mediating PRL and SL release in the goldfish pituitary. PMID- 17920708 TI - IGF-1 controls GLUT3 expression in muscle via the transcriptional factor Sp1. AB - Glucose transporter 3 (GLUT3), while first found in human fetal muscle, is predominantly expressed in brain and neural tissue. By several independent techniques we have previously shown that GLUT3 is expressed in human skeletal muscle cells. The structure of the human GLUT3 gene has not been previously reported nor has there been any evaluation of the 5'-untranslated region (UTR). To this end, we have cloned and sequenced the human GLUT3 gene. Insulin-like growth factor-1 (IGF-1) increased endogenous Glut3 protein in cultured L6 myotubes, and similarly stimulated luciferase activity in a construct of the human GLUT3 5'-UTR linked to a luciferase reporter gene. Actinomycin D, an inhibitor of mRNA synthesis, prevented IGF-1 stimulation of Glut3 protein. Transfection of L6 cells with Sp1 increased Glut3 and augmented IGF-1 stimulation of Glut3 expression. Knockdown of Glut3 expression in cultured L6 muscle cells using small interference RNA (siRNA) specific for Glut3 significantly reduced myocyte glucose uptake. DNAse footprinting and gel shift assays showed Sp1 specifically bound to the human GLUT3 5'-UTR. Substitution mutants of the human GLUT3 5'-UTR luciferase construct indicated that only one of three Sp1 site clusters was involved in IGF-1 action. These data, using both a human GLUT3 5' UTR construct and L6 cells' endogenous promoter, suggest that IGF-1 plays a role in maintaining muscle GLUT3 expression and basal glucose uptake via the transcriptional factor Sp1. PMID- 17920710 TI - Serum carboxy-terminal propeptide of type I procollagen (PIP) is a marker of diastolic dysfunction in patients with early type 2 diabetes mellitus. AB - This study was designed to investigate whether the serum concentration of the carboxy-terminal propeptide of procollagen type I (PIP), a marker of myocardial fibrosis, is related to changes of the ventricular filling dynamics in patients with early type 2 diabetes mellitus (T2DM). The T2DM group had lower mitral and tricuspid E/A ratios than the control group. Serum PIP was higher in patients with T2DM than in controls (131.1+/-45.6 vs. 109.3+/-32.5 ng/mL, p=0.039). A-Ar, an estimate of passive diastolic function, was inversely related to serum PIP levels in T2DM (r=-0.42, p=0.03). These results show a relation between LV diastolic function and serum PIP levels in early T2DM. PMID- 17920709 TI - Myocardial fibrosis in a patient with apical hypertrophic cardiomyopathy detected by delayed-enhanced magnetic resonance imaging. AB - We report a case of apical hypertrophic cardiomyopathy (HCM) presenting with non sustained ventricular tachycardia. Whole-heart magnetic resonance imaging (MRI) showed normal coronary arteries and confirmed the diagnosis of apical HCM. First pass gadolinium delayed-enhanced MRI showed local delayed enhancement in the endomyocardium in the hypertrophic lesion. The presence of perfusion defect and delayed enhancement may be a marker of a high-risk patient with apical HCM. PMID- 17920711 TI - Apical ballooning syndrome during treatment with a vascular endothelial growth factor receptor antagonist. AB - Apical ballooning syndrome (ABS) describes acute regional myocardial dysfunction and has a strong association with emotional stress and female sex. Whilst catecholamine excess has been described in this condition, the precise etiology remains elusive. We report the atypical case of a 61 year old male who developed ABS in the absence of a clear precipitant. His concurrent treatment with a vascular endothelial growth factor (VEGF) receptor antagonist may provide an insight into the pathogenesis of this enigmatic condition. We present a biologically plausible explanation as to why VEGF antagonism may have an important role through its modulation of nitric oxide and catecholamine effects. This hypothesis may also provide an important insight into the cardiovascular toxicities associated with this class of drug. In addition, we report the success of treatment with a beta blocker in ABS complicated by a severe left ventricular outflow tract obstruction. PMID- 17920712 TI - Transient myocardial bridging of the left anterior descending coronary artery in acute inferior myocardial infarction. AB - We observed transient myocardial bridging of left anterior descending coronary artery (LAD) in 18.75% (12 of the total 64) of the patients during acute inferior myocardial infarction (MI). Myocardial bridging occurred only in the acute phase of inferior MI and not in the chronic phase. In the acute phase of inferior MI, compensatory hypercontraction of the anterior wall is assumed to occur in response to the decrease in the movement of the infarct-related walls. In the chronic phase, disappearance of the myocardial bridging observed due to the resolution of compensatory anterior wall hypercontraction, as a result of the reperfusion of infarct-related coronary artery. Most of the myocardial bridges seen in autopsy series are not seen angiographically. Variation at angiography may in part be attributable to small and thin bridges causing little compression. Adrenergic stimulation or afterload reduction by nitroglycerin facilitates diagnosis of myocardial bridging by increasing coronary compression. Both of these conditions are almost always present in acute MI. We concluded that transient myocardial bridging of LAD can be observed in some patients with acute inferior MI during acute stage. PMID- 17920713 TI - Pseudo aneurysm due to dehiscence of a Bentall conduit resulting in systolic aortic compression. PMID- 17920714 TI - How much uncompensated care do doctors provide? AB - The magnitude of provider uncompensated care has become an important public policy issue. Yet existing measures of uncompensated care are flawed because they compare uninsured payments to list prices, not to the prices actually paid by the insured. We address this issue using a novel source of data from a vendor that processes financial data for almost 4000 physicians. We measure uncompensated care as the net amount that physicians lose by lower payments from the uninsured than from the insured. Our best estimate is that physicians provide negative uncompensated care to the uninsured, earning more on uninsured patients than on insured patients with comparable treatments. Even our most conservative estimates suggest that uncompensated care amounts to only 0.8% of revenues, or at most $3.2 billion nationally. These results highlight the important distinction between charges and payments, and point to the need for a re-definition of uncompensated care in the health sector going forward. PMID- 17920715 TI - Five year results of a randomized trial comparing hyperfractionated to conventional radiotherapy over four weeks in locally advanced head and neck cancer. AB - BACKGROUND AND PURPOSE: Fractionation strategies delivered over 4 weeks are of clinical and radiobiological interest because treatment is completed before radiotherapy (RT) induced clonogen proliferation commences in earnest approximately 3 to 4 weeks into a course of RT. We wished to test the clinical hypothesis that an increased total dose delivered over 4 weeks with smaller than standard doses per fraction in locally advanced squamous cell carcinoma (SCC) may result in relative protection of late responding tissues and an increased tumor control compared to a conventional daily course in the same overall time. MATERIALS AND METHODS: Between 1988 and 1995 a randomized controlled trial employing RT alone was undertaken at the Princess Margaret Hospital that included 331 eligible patients with T3 or T4 N0 or any N-positive oropharynx, hypopharynx, or larynx primary SCC. RT was randomly assigned to one of two 4 week schedules, either 51 Gy in 20 equal daily fractions, termed conventional fractionation (CF), or 58 Gy in 40 equal fractions given twice per day as a hyperfractionated (HF) experimental arm. RESULTS: The 5-year local relapse rate was reduced in the HF (41%) compared to the CF arm (49%). This difference was marginally not significant (p=0.082) when the effect was not adjusted. When the effect of the treatment was adjusted by Cox model for clinical factors that included N category, ECOG performance status, site of disease, T-category, age, hemoglobin, and gender the HF achieved a significant effect (p=0.02). Survival (40% vs. 30%) was also improved with HF compared to CF arm. This difference was only marginally not significant (p=0.069) but again achieved statistical significance when the model was adjusted for clinical factors (p=0.01). Similar results were observed for disease free survival. Although reversible acute toxicity was increased with HF, the overall 5-year rate of grade 3 and 4 late toxicity for the CF was 10.5% compared to 7.7% in the higher dose HF arm. CONCLUSIONS: HF delivered in 4 weeks permits enhanced RT doses achieving improved tumor control, without increased late toxicity, compared to daily fractionated radiotherapy in the same overall time. PMID- 17920716 TI - Different response between production of free radicals induced by central and peripheral administration of interleukin-1beta in conscious rats. AB - We examined whether central or peripheral administration of interleukin-1beta (IL 1beta) might change levels of nitric oxide (NO) and hydroxyl radical (*OH) in the medial prefrontal cortex (mPFC). Extracellular levels of NO metabolites (NOx(-)) and 2,3-dihydroxybenzoic acid (2,3-DHBA), as a marker of *OH production, were determined with an in vivo microdialysis technique in conscious rats. In the mPFC, central administration of IL-1beta into the mPFC resulted in dose-dependent increases in levels of both NOx(-) and 2,3-DHBA. In contrast, peripheral administration of IL-1beta significantly increased NOx(-) levels but not 2,3-DHBA levels. Perfusion of Mn(III) tetrakis (4-benzoic acid) porphyrin chloride, a superoxide (O(2)(-)) dismutase mimic, into the mPFC reduced the increases in levels of 2,3-DHBA induced by centrally administered IL-1beta, but enhanced the increases in levels of NOx(-) induced by centrally administered IL-1beta. The present results show a different response in free radical productions in the mPFC between central and peripheral administration of IL-1beta. This finding should be useful for our understanding of the response of NO and free radicals such as *OH and O(2)(-) in the mPFC after central and peripheral administration of IL-1beta. PMID- 17920717 TI - Validation of the (GTG)(5)-rep-PCR fingerprinting technique for rapid classification and identification of acetic acid bacteria, with a focus on isolates from Ghanaian fermented cocoa beans. AB - Amplification of repetitive bacterial DNA elements through the polymerase chain reaction (rep-PCR fingerprinting) using the (GTG)(5) primer, referred to as (GTG)(5)-PCR fingerprinting, was found a promising genotypic tool for rapid and reliable speciation of acetic acid bacteria (AAB). The method was evaluated with 64 AAB reference strains, including 31 type strains, and 132 isolates from Ghanaian, fermented cocoa beans, and was validated with DNA:DNA hybridization data. Most reference strains, except for example all Acetobacter indonesiensis strains and Gluconacetobacter liquefaciens LMG 1509, grouped according to their species designation, indicating the usefulness of this technique for identification to the species level. Moreover, exclusive patterns were obtained for most strains, suggesting that the technique can also be used for characterization below species level or typing of AAB strains. The (GTG)(5)-PCR fingerprinting allowed us to differentiate four major clusters among the fermented cocoa bean isolates, namely A. pasteurianus (cluster I, 100 isolates), A. syzygii- or A. lovaniensis-like (cluster II, 23 isolates), and A. tropicalis like (clusters III and IV containing 4 and 5 isolates, respectively). A. syzygii like and A. tropicalis-like strains from cocoa bean fermentations were reported for the first time. Validation of the method and indications for reclassifications of AAB species and existence of new Acetobacter species were obtained through 16S rRNA sequencing analyses and DNA:DNA hybridizations. Reclassifications refer to A. aceti LMG 1531, Ga. xylinus LMG 1518, and Ga. xylinus subsp. sucrofermentans LMG 18788(T). PMID- 17920718 TI - Response to interferon-alpha treatment correlates with recovery of blood plasmacytoid dendritic cells in children with chronic hepatitis B. AB - BACKGROUND/AIMS: To investigate whether dendritic cell changes are associated with the efficacy of interferon-alpha treatment we longitudinally analyzed circulating dendritic cells in children chronically infected with hepatitis B virus (HBV) undergoing interferon-alpha treatment. METHODS: Thirty-one children with chronic hepatitis B (CHB) received interferon-alpha antiviral treatment for 52 weeks. Myeloid and plasmacytoid dendritic cell (pDCs) frequency and function were analyzed at weeks 0, 2, 12, 24, 36 and 52 in 22 CHB patients. RESULTS: All patients exhibited an initially rapid decrease of circulating pDC numbers and CpG induced endogenous interferon-alpha production within 2 weeks of interferon-alpha treatment. Subsequently, all responders displayed a continuous increase of both pDC numbers and function peaking around week 12. These responses were consequently accompanied by viral clearance, hepatitis B e antigen seroconversion, and the improvement of circulating myeloid dendritic cells and type 1T helper cytokine levels. However, non-responders lacked these sequential responses compared with responders. CONCLUSIONS: pDCs may actively correlate with interferon-alpha therapy-induced viral clearance in pediatric patients with CHB. The recovery of blood pDC number and function may represent a prognostic marker for favourable response to interferon-alpha treatment in chronic hepatitis B. PMID- 17920719 TI - Scanning probe microscopy in the field of drug delivery. AB - The scanning probe microscopes (SPMs) are a group of powerful surface sensitive instruments which when used complimentarily with traditional analytical techniques can provide invaluable, definitive information aiding our understanding and development of drug delivery systems. In this review, the main use of the SPMs (particularly the atomic force microscopy (AFM)) and their successes in forwarding drug delivery are highlighted and categorised into two interlinked sections namely, preformulation and formulation. SPM in preformulation concentrates on applications in pharmaceutical processes including, crystal morphology and modification, discriminating polymorphs, drug dissolution and release, solid state stability and interaction. The ability of the AFM to detect forces between different surfaces and at the same time to operate in liquids or controlled humidity and defined temperatures has also been particularly useful in the study of drug delivery. In formulation, the use of SPMs in different drug delivery systems is discussed in light of different host entry routes. PMID- 17920720 TI - Disulfide bridge based PEGylation of proteins. AB - PEGylation is a clinically proven strategy for increasing the therapeutic efficacy of protein-based medicines. Our approach to site-specific PEGylation exploits the thiol selective chemistry of the two cysteine sulfur atoms from an accessible disulfide. It involves two key steps: (1) disulfide reduction to release the two cystine thiols, and (2) bis-alkylation to give a three-carbon bridge to which PEG is covalently attached. During this process, irreversible denaturation of the protein does not occur. Mechanistically, the conjugation is conducted by a sequential, interactive bis-alkylation using alpha,beta unsaturated-beta'-mono-sulfone functionalized PEG reagents. The combination of: - (a) maintaining the protein's tertiary structure after reduction of a disulfide, (b) bis-thiol selectivity of the PEG reagent, and (c) PEG associated steric shielding ensure that only one PEG molecule is conjugated at each disulfide. Our studies have shown that peptides, proteins, enzymes and antibody fragments can be site-specifically PEGylated using a native and accessible disulfide without destroying the molecules' tertiary structure or abolishing its biological activity. As the stoichiometric efficiency of our approach also enables recycling of any unreacted protein, it offers the potential to make PEGylated biopharmaceuticals as cost-effective medicines. PMID- 17920721 TI - Apoptosis in pyrogallol-treated Calu-6 cells is correlated with the changes of intracellular GSH levels rather than ROS levels. AB - We investigated the involvement of glutathione (GSH) and reactive oxygen species (ROS) such as H2O2 and O2-* in the deaths of pyrogallol-treated Calu-6 cells. Pyrogallol inhibited the growth of Calu-6 cells with an IC50 of approximately 50 microM. Levels of intracellular H2O2 were not altered or were decreased in pyrogallol-treated Calu-6 cells at 72 h. However, levels of O2*- were increased. Treatment with pyrogallol also reduced the intracellular GSH content. The activity of SOD was down-regulated, but the activity of catalase was up-regulated by pyrogallol at 72 h. ROS scavengers, including Tempol, Tiron, Trimetazidine, and N-acetylcysteine (NAC), did not reduce the levels of the intracellular O2*-. Tempol showing the recovery of GSH depletion in pyrogallol-treated cells significantly prevented apoptosis, while Tiron prevented the loss of mitochondrial transmembrane potential (DeltaPsi(m)). In contrast, treatment with NAC showing an increased effect on O2*- levels and depletion of GSH intensified pyrogallol-induced apoptosis. In addition, treatment with SOD and catalase significantly prevented the loss of mitochondrial transmembrane potential (DeltaPsi(m)) in pyrogallol-treated Calu-6 cells. However, only catalase showing a decreased effect on O2*- levels and depletion of GSH prevented pyrogallol induced apoptosis. Taken together, apoptosis in pyrogallol-treated Calu-6 cells is correlated with the changes of intracellular GSH levels rather than ROS levels. PMID- 17920722 TI - ALDH1A1 and ALDH3A1 expression in lung cancers: correlation with histologic type and potential precursors. AB - We hypothesize that aldehyde dehydrogenase (ALDH) isozymes may be upregulated in lung tissue as a result of exposure to carcinogenic aldehydes found in cigarette smoke. To investigate this hypothesis, we studied the expression of two ALDH isozymes in lung cancer from patient samples and its relationship to the history of cigarette smoking. Immunohistochemical staining for ALDH1A1 and ALDH3A1 was performed on archival specimens from control patients without lung cancer, and patients with one of the primary lung cancers: squamous cell cancer (SCCA), adenocarcinoma (AdenoCA), and small cell lung cancer (SCLC). An overall score was obtained for each sample based upon multiplying the staining intensity (0-3) and the extensiveness (0-100%). Mean+/-S.E.M. for each experimental group was calculated and compared. Our results indicate a significantly higher level of expression of ALDH1A1 and ALDH3A1 in SCCA (155+/-19 and 162+/-17, respectively) and AdenoCA (116+/-12 and 107+/-10) than SCLC (39+/-11 and 42+/-12) (P<0.01). Atypical pneumocytes demonstrated significantly higher levels of expression of ALDH1A1 and ALDH3A1 than normal pneumocytes (a normal counterpart of AdenoCA), which is suggestive of up regulation during malignant transformation to AdenoCA. A subset analysis of all samples studied revealed increased expression of ALDH1A1 (P=0.055) and ALDH3A1 (P=0.0093) in normal pneumocytes of smokers (n=32) in comparison to those of non-smokers (n=17). Non-small cell lung cancer (NSCLC) express very high levels of ALDH1A1 and ALDH3A1 in comparison with SCLC, elevated expression of both enzymes may be associated with malignant transformation to AdenoCA, and cigarette smoking seems to result in increased expression of these enzymes in normal pneumocytes. PMID- 17920723 TI - Phase II trial of thalidomide with chemotherapy and as maintenance therapy for patients with poor prognosis small-cell lung cancer. AB - BACKGROUND: Despite the high response rates achieved following standard chemotherapy for small-cell lung cancer (SCLC), the majority of patients will subsequently die from disease progression. MATERIALS AND METHODS: We examined the efficacy and toxicity of thalidomide, an anti-angiogenic agent, in combination with carboplatin and etoposide and as maintenance therapy in patients with untreated SCLC. Twenty-five chemotherapy-naive patients with extensive disease (ED) or limited disease (LD) SCLC were enrolled in a single-arm phase II study. Carboplatin and etoposide were given every 3 weeks for 6 cycles with concurrent thalidomide 100mg orally daily. The treatment with thalidomide was continued as maintenance for up to 2 years. RESULTS: Median progression free and overall survival were 8.3 months and 10.1 months, respectively. One-year survival was 40% and the 1-year progression-free survival was 36%. The overall response rate was 68% (95% CI 46-85%) with four complete remissions (20%) and 13 partial remissions (48%). We observed no increase in chemotherapy related toxicity. Thalidomide was well-tolerated and median time on thalidomide treatment was 7.6 months. CONCLUSION: Concurrent thalidomide with chemotherapy followed by maintenance thalidomide appears to be well tolerated. The results on tumour response rate and survival led us to initiate a randomised phase III trial in the United Kingdom. PMID- 17920724 TI - Lymph node staging by means of positron emission tomography is less accurate in non-small cell lung cancer patients with enlarged lymph nodes: analysis of 1,145 lymph nodes. AB - BACKGROUND: Despite documented superiority of integrated positron emission tomography-computerized tomography (PET-CT) over computerized tomography (CT) in lymph node staging in non-small cell lung cancer, little is known about the sensitivity, specificity and accuracy of integrated PET-CT among enlarged lymph nodes. We sought to assess if PET-CT is uniformly accurate among enlarged and non enlarged lymph nodes. METHODS: A retrospective review of 206 consecutive patients with histologically proven non-small cell lung cancer who underwent resection and/or mediastinoscopy in our centre over 30 months period was carried out. All these patients had pre-operative staging with integrated PET-CT as an adjunct to chest CT prior to resection and/or mediastinoscopy. Diabetic patients (BM>or=8.0 mmol/l) and those who received neo-adjuvant chemotherapy were excluded. The pathological results of all these cases were reviewed and correlated with those on CT and integrated PET-CT. RESULTS: The sensitivity, specificity, accuracy, positive and negative predictive values were higher in integrated PET-CT than CT alone in all lymph nodes, whether N1 or N2. When lymph nodes were stratified by size, the sensitivity of PET-CT was significantly higher among enlarged (>1cm) than non-enlarged (1cm) should be with caution as the specificity of PET-CT is lower and its ability to detect truly negative nodes become reduced. NSCLC patients with enlarged nodes by CT criteria who are PET-CT negative may require cervical mediastinoscopy to rule out metastatic spread to these nodes. Prospective studies are warranted. PMID- 17920726 TI - Characterization of subsoil heterogeneity, estimation of grain size distribution and hydraulic conductivity at the Krauthausen test site using Cone Penetration Test. AB - A Cone Penetration Test (CPT) survey with a high spatial resolution was performed in order to investigate the stratigraphy as well as the spatial variability of various soil properties of the Krauthausen test site. Analyses of the CPT measurements showed the subsurface to be dominated by a planar layered structure. Variogram analysis of the various CPT parameters disclosed that within each layer the soil properties have an anisotropic spatial correlation structure. A correlation analysis of the measured CPT data and co-located grain size distributions from soil samples was performed. Since the correlation coefficients were greater equal to 0.7, a reliable empirical relationship between the data sets could be developed. Based on this empirical relationship grain size distributions were estimated at CPT locations. The statistical processing of estimated and measured grain size distributions with respect to their spatial correlation structure disclosed good agreement between the data sets. The estimated grain size distributions from CPT data were used to estimate the hydraulic conductivity in the aquifer. The results provide detailed information of the spatial heterogeneity of the hydraulic conductivity at Krauthausen test site. The validation of these results, using a prior investigation of hydraulic conductivity statistics, suggests the CPT a fast and inexpensive tool for the estimation of three dimensional hydraulic conductivity fields with sufficient accuracy. PMID- 17920725 TI - Gene methylation in pleural mesothelioma: correlations with clinico-pathological features and patient's follow-up. AB - Methylation of tumor suppressor genes is among the most frequent alterations in patients with malignant pleural mesothelioma (MPM). The aim of this study was to analyze the promoter methylation status of four tumor suppressor genes, p15(INK4B), p16(INK4A), RASSF1A and NORE1A in MPM. Samples of 79 MPM patients were analyzed using a methylation-specific PCR method. Associations between methylation status, clinico-pathological parameters (including proliferation index) and overall survival (OS) were examined. The analysis documented methylation in 30 cases (38%). The methylation frequency for individual genes was 19% for p15(INK4B) (n=15), 11.4% for p16(INK4A) (n=9), 20.2% for RASSF1A (n=16) and 5.1% for Nore1A (n=4). In the whole series methylation was associated to an increased proliferation index (P=0.05). In patients treated with extrapleural pneumonectomy, methylated MPM showed a trend to a poorer OS in comparison to unmethylated cases (median OS 16 months vs. 35 months, P=0.06, HR=2.01, 95% CI 0.95-4.30). In the overall population, methylation did not correlate to patient outcome but a trend to an improved survival was detectable in ummethylated MPM treated with extrapleural pneumonectomy. This result suggests the need to select homogeneously treated and staged patients with MPM to address whether their methylation profile may impact on patient's survival. PMID- 17920727 TI - Proline and glycinebetaine enhance antioxidant defense and methylglyoxal detoxification systems and reduce NaCl-induced damage in cultured tobacco cells. AB - Salt stress impairs reactive oxygen species (ROS) and methylglyoxal (MG) detoxification systems, and causes oxidative damage to plants. Up-regulation of the antioxidant and glyoxalase systems provides protection against NaCl-induced oxidative damage in plants. Thiol-disulfide contents, glutathione content and its associated enzyme activities involved in the antioxidant defense and glyoxalase systems, and protein carbonylation in tobacco Bright Yellow-2 cells grown in suspension culture were investigated to assess the protection offered by proline and glycinebetaine against salt stress. Salt stress increased protein carbonylation, contents of thiol, disulfide, reduced (GSH) and oxidized (GSSG) forms of glutathione, and the activity of glutathione-S-transferase and glyoxalase II enzymes, but decreased redox state of both thiol-disulfide and glutathione, and the activity of glutathione peroxidase and glyoxalase I enzymes involved in the ROS and MG detoxification systems. Exogenous application of proline or glycinebetaine resulted in a reduction of protein carbonylation, and in an increase in glutathione redox state and activity of glutathione peroxidase, glutathione-S-transferase and glyoxalase I under salt stress. Neither proline nor glycinebetaine, however, had any direct protective effect on NaCl-induced GSH associated enzyme activities. The present study, therefore, suggests that both proline and glycinebetaine provide a protective action against NaCl-induced oxidative damage by reducing protein carbonylation, and enhancing antioxidant defense and MG detoxification systems. PMID- 17920728 TI - Combining repeated taste exposure and escape prevention: an intervention for the treatment of extreme food selectivity. AB - Repeated taste exposure has been used to introduce novel foods in several settings, but none of these efforts have targeted clinical populations. This study describes an intervention that combines repeated taste exposure and escape prevention in the treatment of extreme food selectivity in two children with autism. Future applications of repeated taste exposure are discussed. PMID- 17920730 TI - Reduction of foot-and-mouth disease (FMD) virus load in nasal excretions, saliva and exhaled air of vaccinated pigs following direct contact challenge. AB - In future, a policy of "vaccinate-to-live" may be included in the repertoire of foot-and-mouth disease (FMD) control measures and in support of this approach, we have investigated the hypothesis that vaccine-induced reduction in virus replication and excretion from pigs can be correlated to the severity of clinical signs of FMD by measuring excretion of virus in natural secretions and aerosols. The other aims of this study were to verify the existence of sub-clinical infection in vaccinated pigs, to evaluate the correlation between this and seroconversion to foot-and-mouth disease virus (FMDV) non-structural protein antibodies and to re-examine the occurrence of FMDV persistence in the oro pharynx of pigs. Therefore, pigs were vaccinated (O1 Manisa) and challenged (O1 UKG) in a manner calculated to produce a broad range of clinical outcomes and were monitored for a minimum of another 33 days post-challenge. Eighty-one percent of the early (10 days vaccinated) challenged pigs and 25% of the late (29 days vaccinated) challenged pigs were clinically infected and all other vaccinated pigs were sub-clinically infected. Although vaccination could not provide complete clinical or virological protection, it reduced the severity of the disease, virus excretion and production of non-structural FMDV antibodies in vaccinated and subsequently infected pigs. As hypothesised, vaccine-induced reduction of virus replication and excretion was found to be correlated to the severity of clinical disease. RNA copies, but no live virus was detected from the pharyngeal and soft palate tissues of a minority of vaccinated and infected pigs beyond the acute stage of the infection. PMID- 17920732 TI - Hepatitis B seroprevalence and anamnestic response amongst Taiwanese young adults with full vaccination in infancy, 20 years subsequent to national hepatitis B vaccination. AB - The long-term protective effect of hepatitis B virus (HB) vaccination against HB infection and the necessity for routine booster vaccination in young-adult age subsequent to full HB immunization at birth remain issues of some debate currently. This study is aimed at evaluating the seroprevalence of HB infection and the response to HB booster vaccination amongst young-adult university students who had previously undergone full vaccination during their infancy. Eight hundred and forty-three subjects (mean age 18.7+/-0.4 years), 492 males and 351 females, with a complete HB vaccination during infancy were enrolled into this study. The prevalence of natural HB infection, chronic HB-carrier status, and HB-naive group was, respectively, 4.1%, 1.4%, and 62.3%. Amongst 316 study subjects who were naive to HB infection and had received one HB booster at time of university entrance health examination, 49.6%, 91.4%, and 97.5% of the participants with a serum anti-HBs level <0.1, 0.1 to <1.0 and 1.0 to <10.0mIU/mL prior to the booster vaccination, respectively, developed an anamnestic response (i.e., >/=10mIU/mL) to a booster dose of HB vaccine. Full implementation of national-wide HB vaccination program in 1986 has significantly reduced the incidence of HB infection and associated carrier rate in Taiwan. Approximately three-quarter of the subjects who were naive to HB infection and had received one HB booster demonstrated an anamnestic response to a booster HB vaccine. The higher the anti-HBs titers remained for an individual subsequent to primary vaccination, the greater the anamnestic response observed. Additional long-term follow-up studies are needed for young adults initially vaccinated for HB in their infancy. PMID- 17920731 TI - Induction of HIV-specific functional immune responses by a multiclade HIV-1 DNA vaccine candidate in healthy Ugandans. AB - A phase I randomized, double blind, placebo-controlled trial to assess the immunogenicity of a multiclade HIV-1 DNA plasmid vaccine was conducted in 31 HIV 1-negative Ugandans. Following immunization with DNA at 0, 1, and 2 months, the frequency of HIV-specific immune responses was assessed up to 10 months using a standard chromium release assay (CRA), lymphoproliferative assay (LPA), and antibody dependent cell-mediated cytotoxicity assay (ADCC). Seven of 15 (47%) vaccinees demonstrated CTL activity using the CRA to HIV-1 Env B with responses observed 1 month following the second vaccination and as late as 7 months following complete immunization. Additionally, lymphoproliferative reponses were observed in 14/15 vaccinees against p24. No CTL or LPA responses were observed at baseline or in the placebo group. ADCC activity was minimally induced by DNA vaccination. This study demonstrates that immunization with DNA alone induces CTL and lymphoproliferative responses in a population that will participate in a phase IIb study evaluating HIV-1 DNA priming followed by boosting with a replication-defective recombinant adenovirus vector. PMID- 17920729 TI - Postural sway reduction in aging men and women: relation to brain structure, cognitive status, and stabilizing factors. AB - Postural stability becomes compromised with advancing age, but the neural mechanisms contributing to instability have not been fully explicated. Accordingly, this quantitative physiological and MRI study of sex differences across the adult age range examined the association between components of postural control and the integrity of brain structure and function under different conditions of sensory input and stance stabilization manipulation. The groups comprised 28 healthy men (age 30-73 years) and 38 healthy women (age 34-74 years), who completed balance platform testing, cognitive assessment, and structural MRI. The results supported the hypothesis that excessive postural sway would be greater in older than younger healthy individuals when standing without sensory or stance aids, and that introduction of such aids would reduce sway in both principal directions (anterior-posterior and medial-lateral) and in both the open-loop and closed-loop components of postural control even in older individuals. Sway reduction with stance stabilization, that is, standing with feet apart, was greater in men than women, probably because older men were less stable than women when standing with their feet together. Greater sway was related to evidence for greater brain structural involutional changes, indexed as ventricular and sulcal enlargement and white matter hyperintensity burden. In women, poorer cognitive test performance related to less sway reduction with the use of sensory aids. Thus, aging men and women were shown to have diminished postural control, associated with cognitive and brain structural involution, in unstable stance conditions and with diminished sensory input. PMID- 17920734 TI - Transfer of (40)K, (238)U, (210)Pb, and (210)Po from soil to plant in various locations in south of Syria. AB - Transfer factors of (40)K, (238)U, (210)Pb, and (210)Po from soil to some agriculture crops in various locations in south of Syria (Dara'a and Assuwaydaa districts) have been determined. Soil and vegetable crops (green pepper, cucumber, tomato, and eggplant), legumes crops (lentil, chickpea, and broad bean), fruit trees (apple, grape, and olives) and cereals (barley and wheat) were collected and analyzed for (238)U, (210)Pb, and (210)Po. The results have shown that higher transfer factors (calculated as Bqkg(-1) dry wt. plant material per Bqkg(-1) dry wt. soil) for (210)Po, (210)Pb and (238)U were observed in vegetable leaves than fruits and cereals leaves; the highest values of transfer factor (TF) for (238)U were found to be 0.1 for straw of chickpea. Transfer factors for (210)Po varied between 2.8x10(-2) and 2 in fruits of eggplant and grain of barley, respectively. In addition, several parameters affecting transfer factors of the radionuclides were evaluated. The results can be considered as base values for TF of natural radionuclides in the region. PMID- 17920733 TI - Caprine herpesvirus 1 vaccine with the LTK63 mutant as a mucosal adjuvant induces strong protection against genital infection in goats. AB - Caprine herpesvirus 1 provides a unique virus-animal model to investigate potential tools applicable for the therapy and prophylaxis of genital herpesvirus infections of humans. In order to evaluate the efficacy of mucosal immunization in the goat model, an inactivated CpHV-1 vaccine was adjuvated with the enzymatically inactive mutant of the heat-labile enterotoxin of Escherichia coli, LTK63, and used to immunize goats by the vaginal route, by administering two doses at a 3-week interval. The mucosal vaccine was safe, as neither local nor systemic reactions were associated with the vaccine administration. The vaccinated animals displayed high levels of secretory IgA and were significantly protected after challenge with the virulent CpHV-1 strain, with marked decrease in virus shedding, while the unvaccinated goats were not. These findings suggest that mucosal immunization is potentially exploitable in the control of genital infection by herpesviruses. PMID- 17920735 TI - Radionuclides and metals in freshwater mussels of the upper South Alligator River, Australia. AB - During an inspection of the old Rockhole Mine area in Kakadu National Park in 1999, it was found that a small amount of tailings from the former South Alligator uranium (U) mill had been uncovered by wet season rain and road works. Samples of sediment, water and freshwater mussels, Velesunio angasi, were collected from the South Alligator River, near and at the confluence of Rockhole Mine Creek, and adjacent to the exposed tailings. The 228Ra/226Ra activity ratios in sediments and mussel tissue indicate a small influence from the tailings and from Rockhole Mine adit water on 226Ra concentrations. The uptake of 226Ra in mussels does not correlate with other alkaline-earth metals. Mussel U concentrations are higher immediately downstream of Rockhole Mine Creek, but there is no noticeable increase in the immediate vicinity of the tailings area. A hypothetical ingestion of 2 kg of mussels from the sites was used to estimate the committed effective dose for a 10-year-old child resulting in a figure of 0.23 mSv per annum, of this total dose, 69% is attributed to 210Po. Only 0.03 mSv per annum can be directly linked to impacts of the tailings. PMID- 17920736 TI - Arthrocentesis in the treatment of loose bodies of the temporomandibular joint associated with synovial chondromatosis. AB - Synovial chondromatosis is a benign disorder of joints of unknown aetiology, characterised by the presence of loose bodies in the articular space. We present a case that affected the temporomandibular joint (TMJ) and was treated with arthrocentesis, which is an efficient, conservative, and inexpensive treatment. PMID- 17920737 TI - Factors predicting the initiation of prenatal care in Mexican women. AB - OBJECTIVE: to describe factors related to the initiation of prenatal care (PNC) among pregnant Mexican women. DESIGN: descriptive correlational. SETTING: public prenatal clinic in Monterrey, Mexico. PARTICIPANTS: 253 pregnant Mexican women aged 13-46 years. MEASUREMENTS AND FINDINGS: structured interviews were conducted to obtain information concerning demographics, reproductive history, current pregnancy, perceptions of benefits and barriers of PNC, negative attitudes towards pregnancy and social support. Late initiation of PNC was reported by 47% of participants. Factors related to PNC initiation were education, perceived benefits or barriers, and negative attitudes towards pregnancy. Women who initiated PNC at an early stage were more likely to live with a partner, had a higher educational level and anticipated desirable personal benefits of PNC. Perceptions of barriers to PNC impeded early initiation of care. Social support did not influence PNC initiation. Negative attitudes towards pregnancy increased the likelihood of late PNC initiation. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: screening for negative attitudes towards pregnancy, partner status, educational level and perceived health-care barriers among pregnant Mexican women is important. Health-care providers may then address these psychosocial risk factors through PNC interventions promoting early initiation of care. PMID- 17920739 TI - Bioavailability assessment of contaminants in soils via respiration and nitrification tests. AB - For the assessment of contaminated soils ecotoxicological tests are used to estimate the bioavailability of contaminants in soil samples. Terrestrial tests reveal the habitat function of soils, and parameters applied in tests involving microorganisms include respiration activity and potential ammonium oxidation. For such tests, the threshold values needed to assess the results have already been established in guidelines ISO 17155 and ISO 15685. In this paper, we discuss about the respiration activity and potential ammonium oxidation results obtained from a wide variety of soils with different physico-chemical properties and levels of contamination. These results show that microbial respiration and potential ammonium oxidation have different sensitivities to various classes of contaminants. We demonstrated that both organic and inorganic contaminants influence potential ammonium oxidation, whereas microbial respiration is predominantly affected by biodegradable organic contaminants. These differences might be useful for more detailed assessments of soil contamination, leading to different recommended actions depending on which parameter is affected. PMID- 17920738 TI - The effect of functional knee brace design and hinge misalignment on lower limb joint mechanics. AB - BACKGROUND: Knee bracing has been shown to alter lower limb joint mechanics, which may protect the anterior cruciate ligament. The effect of brace alignment and brace type, however, remains largely unknown. This study was conducted to determine whether the use of a functional knee brace, the type of brace used or its alignment relative to the knee causes biomechanical alterations to gait. METHODS: Ten healthy participants took part in two walking conditions (aligned brace and misaligned brace) for two different types of brace (sleeve brace with bilateral hinges and hinge-post-shell). A non-braced condition was included as a baseline measure. Three-dimensional kinematics and force platform data were used to calculate the joint intersegmental forces and net joint moments of the ankle, knee and hip. FINDINGS: In comparison to non-braced walking, the shell brace in its aligned position significantly reduced the peak ankle plantarflexor moment. There was a decreased peak knee flexion angle with both the aligned shell and sleeve braces. The shell brace in its aligned position significantly increased peak knee adduction and reduced peak knee internal rotation. INTERPRETATION: In this sample of healthy participants, functional knee bracing failed to alter lower limb mechanics in such a way that would reduce the force transmitted to the anterior cruciate ligament. In addition, although there were brace induced changes in lower limb kinematics with 2cm of distal hinge misalignment, it is unlikely that hinge misalignment of this magnitude is detrimental to an uninjured knee joint during walking. PMID- 17920740 TI - Implementing community interventions for HIV prevention: insights from project ethnography. AB - Global policy on HIV prevention among marginalised populations recommends a community-based approach with participation and mobilisation as central features. The overall aim is to empower individuals and groups to reduce their vulnerability to HIV. Community empowerment initiatives have had mixed results, however, in spite of utilising very similar peer-education based intervention strategies. This is particularly true of community-based interventions in sex work settings. Drawing upon an ethnographic study conducted in the early years of a well-known sex worker initiative-the Sonagachi STD/HIV Intervention Project (SHIP) in Kolkata, India-this paper argues that ethnographic research can illuminate the complex and inter-dependent dynamics of context, practice, agency and power that are specific to a project and shape the course of intervention implementation in ways that may be 'hidden' in conventional techniques of project reporting. Two detailed excerpts of what we refer to as 'private contexts of practice' are presented-focusing upon the complex processes underlying community mobilisation and peer education. We show that the gathering of ethnographic forms of evidence in conjunction with more conventional evaluation measures has two distinct benefits. First, an ethnographic approach is able to capture the play of power through observation of real-time events that involve multiple actors with widely different perspectives, as compared with retrospective accounts from carefully selected project representatives (the usual practice in project evaluations). Second, observation of actual intervention practices can reveal insights that may be hard for project staff to articulate or difficult to pinpoint, and can highlight important points of divergence and convergence from intervention theory or planning documents. PMID- 17920741 TI - Low perceived benefits and self-efficacy are associated with hepatitis C virus (HCV) infection-related risk among injection drug users. AB - Hepatitis C prevention counselling and education are intended to increase knowledge of disease, clarify perceptions about vulnerability to infection, and increase personal capacity for undertaking safer behaviours. This study examined the association of drug equipment sharing with psychosocial constructs of the AIDS Risk Reduction Model, specifically, knowledge and perceptions related to hepatitis C virus (HCV) among injection drug users (IDUs). Active IDUs were recruited between April 2004 and January 2005 from syringe exchange and methadone maintenance treatment programs in Montreal, Canada. A structured, interviewer administered questionnaire elicited information on drug preparation and injection practices, self-reported hepatitis C testing and infection status, and AIDS Risk Reduction Model constructs. Separate logistic regression models were developed to examine variables in relation to: (1) the sharing of syringes, and (2) the sharing of drug preparation equipment (drug containers, filters, and water). Among the 321 participants, the mean age was 33 years, 70% were male, 80% were single, and 91% self-identified as Caucasian. In the multivariable analyses, psychosocial factors linked to syringe sharing were lower perceived benefits of safer injecting and greater difficulty to inject safely. As with syringe sharing, the sharing of drug preparation equipment was associated with lower perceived benefits of safer injecting but also with low self-efficacy to convince others to inject more safely. Interventions should aim to heighten awareness of the benefits of risk reduction and provide IDUs with the skills necessary to negotiate safer injecting with their peers. PMID- 17920742 TI - Understanding feticide: an analytic review. AB - The medical procedure of 'feticide' has been used in clinical practice since the early 1990s in the UK. The procedure constitutes a sensitive aspect of late termination of pregnancy (TOP), an issue that is in itself contentious. The procedure has attracted attention from academic and policy commentators, but recently the medical profession has expressed some uncertainty with respect to the legal position of live birth following TOP, and professional discretion in providing feticide. To understand the meaning of these comments better, we argue that it is helpful to acknowledge the rhetoric that shapes the academic discourse on feticide. In this paper, we review how feticide has been conceptualised within academic discourse, demonstrating that the concept has multiple meanings, some of which could be considered politically charged. We then consider some examples of the comments made about the legal uncertainties of feticide, highlighting assumptions made about the problematic nature of professional discretion. Ultimately, we suggest that a better understanding of the context of feticide is needed to ensure that future research in this area of health care engages adequately with issues of professional discretion. PMID- 17920743 TI - The social epidemiology of tuberculosis in South Africa: a multilevel analysis. AB - Increased risk of tuberculosis is widely recognized to be associated with increased poverty, yet there have been few analyses of the social determinants of tuberculosis, particularly in high-burden settings. We conducted a multilevel analysis of self-reported tuberculosis disease in a nationally representative sample of South Africans based on the 1998 Demographic and Health Survey (DHS). Individual and household-level demographic, behavioral and socioeconomic risk factors were taken from the DHS; data on community-level socioeconomic status (including measures of absolute wealth and income inequality) were derived from the 1996 national census. Of the 13,043 DHS respondents, 0.5% reported having been diagnosed with tuberculosis disease in the past 12 months and 2.8% reported having been diagnosed with tuberculosis disease in their lifetime. In a multivariate model adjusting for demographic and behavioral risk factors, tuberculosis diagnosis was associated with cigarette smoking, alcohol consumption and low body mass index, as well as a lower level of personal education, unemployment and lower household wealth. In a model including individual- and household-level risk factors, high levels of community income inequality were independently associated with increased prevalence of tuberculosis (adjusted odds ratio for lifetime tuberculosis comparing the most unequal quintile to the middle quintile of inequality: 2.37, 95% confidence interval: 1.59-3.53). These results provide novel insights into the socioeconomic determinants of tuberculosis in developing country settings, although the mechanisms through which income inequality may affect tuberculosis disease require further investigation. PMID- 17920744 TI - Treatment of clozapine-associated tardive dyskinesia. PMID- 17920746 TI - Changes in the hepatic gene expression profile in a rat model of chronic ethanol treatment. AB - The purpose of this study was to perform a comprehensive analysis of hepatic gene expression in a standard model of an alcohol-induced fatty liver using the cDNA microarray analysis. Male Sprague-Dawley rats were randomly divided into two groups and were given either an ethanol diet (ED), or a control diet (CD) for eight weeks. The ED rats showed significantly elevated levels of plasma total and HDL cholesterol as well as hepatic cholesterol and triglyceride compared to the pair-fed control rats. Among the 5185 genes on the rat cDNA microarray used in the current study, 74 genes were up-regulated and 108 genes were down-regulated greater than 2.0-fold in the liver of ED rats compared with those in the CD rats. The microarray results were verified by conducting real-time RT-PCR on the fourteen selected genes with varied expression ratios. After clustering the regulated genes based on their biological function, it was found that chronic ethanol consumption regulated mainly the genes implicated in the processes of signal transduction, transcription, immune response, and protein/amino acid metabolism. The microarray results obtained in this study revealed, for the first time, that several genes, including beta-glucuronidase, UDP-glycosyltransferase 1, UDP-glucose dehydrogenase, apoC-III, and gonadotropin-releasing hormone receptor, were regulated by chronic ethanol exposure in the rat liver. PMID- 17920745 TI - Genotoxicity study with special reference to DNA damage by comet assay in fission yeast, Schizosaccharomyces pombe exposed to drinking water. AB - The objective of this study was to investigate genotoxicity, especially DNA damage, in drinking water samples collected from tap by using fission yeast Schizosaccharomyces pombe as a model organism. Generally raw water potabolization is done by treatment with polymeric coagulant, alum, chlorine, etc. In the comet test, highly significant (P<0.001) effects of DNA damage were detected in treated water (tap water) when compared to negative control (raw water) as well as laboratory control (distilled water) samples for both 1 h and 2 h exposure. In the water treatment plant, raw water treatment is done by the process of prechlorination, alum and polymeric coagulant (CatflocT) dosing, postchlorination, filtration and final discharge for consumption. In conclusion it can be stated from the results that chlorinated disinfectant, alum and polymeric coagulant (CatflocT) mixture used in drinking water has a potent cumulative genotoxic effect in the eukaryotic cells and may pose potential genotoxic risk for human health following long-term consumption. PMID- 17920747 TI - Urinary biomarkers of aflatoxin exposure in young children from Egypt and Guinea. AB - Aflatoxins are a major risk factor for hepatocellular carcinoma (HCC), and thus understanding the pattern of aflatoxin exposure in different regions is important in order to develop targeted intervention strategies. Given the early onset of HCC in many countries early life exposures may be important. This study investigated aflatoxin exposure in Egyptian children (n=50, aged 1-2.5 years) by assessing urinary aflatoxin metabolite (AFM(1), AFB(1), AFB(2), AFG(1), AFG(2)) levels. Samples from Guinean children (n=50, aged 2-4 years) were analyzed in parallel providing a comparison to a region of established frequent aflatoxin exposure. Aflatoxins were isolated from urine using C18-cartridges followed by immunoaffinity clean-up, and quantified by HPLC with fluorescence detection. Overall aflatoxins were less frequently present in Egyptian (38%) than Guinean urine samples (86%) (p<0.001), which was particularly related to differences in detection rates of AFM(1) (8% compared to 64%, respectively, (p<0.001)). For AFM(1) the geometric mean level in Guinea (16.3 pg/ml; 95% CI: 10.1, 26.6 pg/ml) was 6-fold higher (p<0.001) than in Egypt (2.7 pg/ml; 95% CI: 2.5, 2.8 pg/ml). Urinary aflatoxins from healthy children in these two regions have not previously been reported, and exposure appears modest in Egypt compared to Guinea. These data suggest that measures to reduce aflatoxin exposure in both regions are important, though particularly in Guinea. PMID- 17920748 TI - Comparative studies on the increase of uterine weight and related mechanisms of cadmium and p-nonylphenol. AB - The research was designed to compare the effect of cadmium and p-nonylphenol on the increase of uterine weight and to study the related mechanisms. It provided basic evidence for us to understand the possible different mechanisms among different EEDs. In this study, both ovaries of 60 Wistar rats (28 days age) were ectomized, and after 21 days recovery, the rats were randomly assigned into six groups and exposed to cadmium (0.12, 1.20mg/kg), NP (100, 200mg/kg), control (sterile PBS), and positive control (17beta-estradiol) per day for 3 days, respectively, then related indexes were detected. The results showed that the increase of uterine weight induced by cadmium was accompanied by the increase of the thickness of luminal epithelium cell and endometrium but the decrease of nuclear/cytoplasm of luminal epithelium cell and endometrium, while the increase of uterine weight induced by p-nonylphenol was accompanied by the increase of the thickness of luminal epithelium cell, endometrium, and myometrium but the decrease of nuclear/cytoplasm of luminal epithelium cell and endometrium. Cadmium could inhibit the positive expression of PCNA while p-nonylphenol prompted it. Exposure to cadmium and NP both could also stimulate phosphorylation of ERK mitogen-activated protein kinases, implying that this signal pathway had an effect on the increase of the uterine weight induced by cadmium and p nonylphenol. The results indicate that cadmium may induce the increase of uterine weight, which is accompanied with toxic effect on endometrium, while NP's effect of the increase of uterine weight is due to cell proliferation of endometrium, the mechanisms of which are the same as estrogen, but they may both activate ERK signal pathway. PMID- 17920749 TI - Emerging roles of ADAM and ADAMTS metalloproteinases in cancer. AB - A disintegrin and metalloproteinases (ADAMs) are a recently discovered family of proteins that share the metalloproteinase domain with matrix metalloproteinases (MMPs). Among this family, structural features distinguish the membrane-anchored ADAMs and the secreted ADAMs with thrombospondin motifs referred to as ADAMTSs. By acting on a large panel of membrane-associated and extracellular substrates, they control several cell functions such as adhesion, fusion, migration and proliferation. The current review addresses the contribution of these proteinases in the positive and negative regulation of cancer progression as mainly mediated by the regulation of growth factor activities and integrin functions. PMID- 17920750 TI - Automatic identification of large collections of protein-coding or rRNA sequences. AB - The number of available genomic sequences is growing very fast, due to the development of massive sequencing techniques. Sequence identification is needed and contributes to the assessment of gene and species evolutionary relationships. Automated bioinformatics tools are thus necessary to carry out these identification operations in an accurate and fast way. We developed HoSeqI (Homologous Sequence Identification), a software environment allowing this kind of automated sequence identification using homologous gene family databases. HoSeqI is accessible through a Web interface (http://pbil.univ lyon1.fr/software/HoSeqI/) allowing to identify one or several sequences and to visualize resulting alignments and phylogenetic trees. We also implemented another application, MultiHoSeqI, to quickly add a large set of sequences to a family database in order to identify them, to update the database, or to help automatic genome annotation. Lately, we developed an application, ChiSeqI (Chimeric Sequence Identification), to automate the processes of identification of bacterial 16S ribosomal RNA sequences and of detection of chimeric sequences. PMID- 17920751 TI - Electrophysiological correlates of category induction: PSW amplitude as an index of identifying shared attributes. AB - Event-related potentials (ERPs) were recorded from 15 healthy adults while they performed a category induction task. Two geometric figures characterized by three dimensions (color, shape, and stripe orientation) were presented simultaneously to subjects who were asked to identify their shared attributes. Pairs of figures sharing common attributes composed the category induction condition and pairs of identical figures composed the non-induction condition. It was hypothesized that non-induction only involves abstracting shared attributes from one stimulus, but category induction involves identifying shared attributes by comparing the two stimuli. The ERP waves elicited by induction and non-induction did not differ on N1, P2 and P3b components, but larger frontal N2 and smaller central-parietal PSW components were elicited by category induction. These results suggest that the processing of category induction is reflected in the PSW 400-650 ms post stimulus. PMID- 17920752 TI - Previable rupture of membranes: effect of amniotic fluid on pregnancy outcome. AB - OBJECTIVE: To evaluate pregnancy outcome and the role of the amount of amniotic fluid (AF) in the prognosis of extremely preterm (<24 weeks) premature rupture of membranes (EPPROM). STUDY DESIGN: Women with EPPROM and on-going pregnancy after 1 week of expectant management were included. EXCLUSION CRITERIA: fetal anomalies, termination of pregnancy and spontaneous recovery of AF within the first week. The effect of the large vertical pocket (LVP) on pregnancy outcome was assessed by a Cox regression model which included three covariates: LVP measurements from rupture to 24 weeks, gestational age at rupture of membranes and sealing procedure. RESULTS: Thirty-seven women were included in the study. The overall survival rate after the neonatal period was 40.5% (15/37) which increased to 62.5% (15/24) in neonates born alive after 24 weeks of gestation. Mean and S.D. of gestational age at rupture of membranes were 19.0 (+/-3.8) weeks. From rupture to 24 weeks, the pooled mean and standard deviation of LVP were 20.5 (+/-15.4)mm. Multivariate analysis showed that the likelihood for neonate survival increased by 2.7 (95% CI 1.45-4.65) for each 5mm of LVP during the follow-up from rupture to 24 weeks. After controlling for AF amount, neither the gestational age at rupture nor the sealing procedure showed any significant effect on pregnancy outcome. CONCLUSION: Although the prognosis of EPPROM is poor overall, survival improves as the amount of AF before 24 weeks increases. PMID- 17920753 TI - Thymus repopulation after allogeneic reconstitution in hematological malignancies. AB - OBJECTIVE: Active vaccination in the allogeneically reconstituted tumor-bearing host essentially requires donor T-cell tolerance. To create a basis for vaccination in the allogeneically reconstituted, lymphoma-bearing host, we elaborate a reconstitution protocol that supports thymus repopulation and tolerance induction. METHODS: Myeloreductively conditioned, lymphoma-bearing mice were vaccinated after reconstitution with hematopoietic progenitor cells. Readout systems included recovery of donor-derived T cells, graft vs host disease (GVHD), anti-host and anti-lymphoma cytotoxicity, as well as tumor growth rate and tumor rejection. RESULTS: In tumor-free mice, myeloreductive conditioning, together with natural killer cell depletion of the host and transfer of T cell-depleted bone marrow cells, allows reconstitution without severe GVHD. However, in hematological malignancies, donor-derived T-progenitor cells hardly immigrated into the thymus. As a consequence, the frequency of severe GVHD was significantly increased, which prohibited active vaccination. Thymus repopulation became improved by strengthening myeloreductive conditioning; by supporting thymocyte expansion via interleukin-7; and, most strongly, by a small dose of donor-derived CD4(+)CD8(+) thymocytes, which preferentially homed into the thymus. Active vaccination, in combination with this reconstitution protocol, did not strengthen GVHD, but significantly improved survival time and survival rate of lymphoma bearing mice. CONCLUSION: The negative impact of hematological malignancies on thymus repopulation and central tolerance induction can, at least in part, be corrected by application of a small number of donor-derived T-progenitor cells. PMID- 17920754 TI - The association of JAK2V617F mutation and leukocytosis with thrombotic events in essential thrombocythemia. AB - OBJECTIVES: The Janus kinase 2 mutation, JAK2 (V617F), and megakaryocytic mutations, MPL (W515L/K), have been identified and correlated with a subtype of essential thrombocythemia (ET) patients. We investigated the frequency of mutations in ET patients and analyzed the relationship with their clinical features. METHODS: Fifty-three ET patients were enrolled in the study. The amplification refractory mutation system was applied for the mutation survey of the JAK2V617F, while the polymerase chain reaction with sequencing was used for the mutation survey of MPLW515L/K. RESULTS: Thirty-five (66%) patients harboring the JAK2 (V617F) mutation, including 3 homozygous and 32 heterozygous changes, but no MPLW515L/K mutation, were found. During follow-up, 17 (32.1%) patients suffered from documented thrombotic events, with 15 having JAK2V617F mutations. Statistical analysis showed that patients with the JAK2 mutation had significantly higher leukocytes, hemoglobin level, and thrombotic event (p = 0.043, p = 0.001, and p = 0.029, respectively). Thrombotic events were also significantly correlated with leukocytosis and older age. CONCLUSIONS: The JAK2V617F mutation was noted in a certain population of ET patients and correlated with leukocytosis, high hemoglobin level, and thrombosis. Therefore, detection of the JAK2V617F mutation can affect not only the diagnosis, but also the management of ET patients. PMID- 17920756 TI - MOZ and MOZ-CBP cooperate with NF-kappaB to activate transcription from NF-kappaB dependent promoters. AB - OBJECTIVE: Monocytic zinc finger (MOZ) maintains hematopoietic stem cells and, upon fusion to the coactivator CREB-binding protein (CBP), induces acute myeloid leukemia (AML). Leukemic stem cells in AML often exhibit excessive signal dependent activity of the transcription factor nuclear factor (NF)-kappaB. Because aberrant interaction between NF-kappaB and coactivators represents an alternative mechanism for enhancing NF-kappaB activity, we evaluated whether MOZ and MOZ-CBP cooperate with NF-kappaB to activate transcription from NF-kappaB dependent promoters. METHODS: The ability of MOZ, MOZ mutants, and MOZ-CBP to enhance expression of NF-kappaB-dependent promoters was tested in reporter studies. The interaction between MOZ and NF-kappaB was evaluated by both coimmunoprecipitation and glutathione S-transferase pulldown assays. RESULTS: MOZ activates transcription from the NF-kappaB-dependent interleukin-8 promoter; interestingly, this effect is markedly enhanced by CBP. Although MOZ has less potent transcriptional activity than MOZ-CBP, both proteins cooperate with steroid receptor coactivator-1 to activate transcription. MOZ also induces multiple NF-kappaB-dependent viral promoters. Importantly, MOZ associates in a protein complex with the p65 subunit of NF-kappaB and interacts directly with p65 in vitro. Transcriptional activity of MOZ requires its C-terminal domain, which is absent from MOZ-CBP, indicating that the transcriptional activity of MOZ-CBP derives from its CBP sequence. CONCLUSIONS: MOZ interacts with the p65 subunit of NF-kappaB and enhances expression of NF-kappaB-dependent promoters. The more potent transcriptional activity of MOZ-CBP derives from its CBP sequence. Thus, interaction between NF-kappaB and MOZ-CBP may play an important role in the pathogenesis of certain acute myeloid leukemias. PMID- 17920755 TI - Phenotypic variations and new mutations in JAK2 V617F-negative polycythemia vera, erythrocytosis, and idiopathic myelofibrosis. AB - OBJECTIVE: The chronic myeloproliferative disorders (MPD), polycythemia vera (PV), essential thrombocytosis, and idiopathic myelofibrosis (IMF), are characterized by a spectrum of clinical features and linked by common genetic lesions in JAK2 and MPL. However, the clinical phenotypes in genetically undefined MPD patients are similar to those patients with JAK2 and MPL lesions. We, therefore, sought to determine whether there were JAK2 or MPL lesions in a well-defined, JAK2 V617F-negative MPD cohort, and to determine if clinical associations could be identified based on variations identified in these genes. METHODS: We examined the JAK2 and MPL genes in JAK2 V617F-negative PV, IMF, and idiopathic erythrocytosis patients for sequence variations. RESULTS: We identified two previously unrecognized JAK2 mutations and three previously unrecognized MPL mutations in JAK2 V617F-negative PV, erythrocytosis, and IMF patients. We identified JAK2 exon 12 lesions in 30% of JAK2 V617F-negative PV patients, and either JAK2 V617F or JAK2 exon 12 lesions in 9% of erythrocytosis patients. In IMF, in addition to the MPL gene mutation, W515K, we identified three additional mutations: 204P and two intervening sequence transitions, IVS 11/12 and 10/11. CONCLUSIONS: While the clinical phenotype of JAK2 exon 12 lesions in the MPD was predominantly erythroid, there was significant disease spectrum overlap between JAK2 V617F and JAK2 exon 12 mutations. By contrast, MPL gene mutations were not associated with erythrocytosis, but segregated primarily with the phenotypes of thrombocytosis, extramedullary disease, myelofibrosis, and osteosclerosis. PMID- 17920757 TI - Unmanipulated HLA 2-3 antigen-mismatched (haploidentical) bone marrow transplantation using only pharmacological GVHD prophylaxis. AB - OBJECTIVE: The incidence of severe graft-vs-host disease (GVHD) in unmanipulated human leukocyte antigen (HLA) 2-3 antigen-mismatched bone marrow transplantation (BMT) using cyclosporine and methotrexate as GVHD prophylaxis is 80% to 90%. We investigated whether pharmacological GVHD prophylaxis consisting of four drugs, including a steroid, effectively suppressed GVHD in this transplantation setting. MATERIALS AND METHODS: Thirty patients who had hematologic malignancies at an advanced stage or with poor prognosis underwent allogeneic BMT using a myeloablative preconditioning regimen consisting of cyclophosphamide (60 mg/kg x 2), total body irradiation (8-10 Gy), and fludarabine (30 mg/m(2) x 4) with or without cytosine arabinoside (2 g/m(2) x 4), and GVHD prophylaxis consisting of a combination of tacrolimus, methotrexate, mycophenolate mofetil, and methylprednisone (2 mg/kg). Early therapeutic intervention for GVH reaction or grade I GVHD was performed, and steroid was slowly tapered. RESULTS: All patients achieved donor-type engraftment. Neutrophil (>0.5 x 10(9)/L) and platelet (>20 x 10(9)/L) engraftment was achieved on day 13 and on day 30, respectively. Seventeen patients (56.7%) had no GVHD. Eleven patients (36.7%) developed grade II-III acute GVHD. Seven patients (23.3%) died of transplant-related toxicity, including fungal or viral infections and thrombotic microangiopathy, and four patients died of disease progression. Estimated relapse rate at 3 years was only 20.9%. The probability of survival at 3 years was 49.9%. CONCLUSIONS: These data suggest that, in unmanipulated HLA-haploidentical allogeneic BMT, this GVHD prophylactic regimen, which includes methylprednisolone 2 mg/kg, and early therapeutic intervention for GVH reaction suppress the incidence of severe GVHD to an acceptable level, while preserving the graft-vs-leukemia effect. PMID- 17920758 TI - Vectors selected from adeno-associated viral display peptide libraries for leukemia cell-targeted cytotoxic gene therapy. AB - OBJECTIVE: For acute myeloid leukemia (AML), gene therapy may be used to treat patients refractory to conventional chemotherapy. However, availability of vectors sufficiently and specifically transducing this cell type is very limited. METHOD: Here we report the selection of capsid-modified adeno-associated viral (AAV) vectors targeting Kasumi-1 AML cells by screening random AAV displayed peptide libraries. RESULTS: The peptide inserts of the enriched capsid mutants share a common sequence motif. The same motif was selected in an independent library screening on HL-60 AML cells. Recombinant targeted vectors displaying the selected peptides transduced the target leukemia cells they have been selected on up to 500-fold more efficiently compared to AAV vectors with control peptide inserts. One of the selected clones (NQVGSWS) also efficiently transduced all members of a panel of four other AML cell lines. Binding and blocking experiments showed that NQVGSWS binding to leukemia cells is independent of the wild-type AAV 2 receptor heparin sulfate proteoglycan. Transduction assays on a panel of hematopoietic and nonhematopoietic cell lines showed that the NQVGSWS capsid was able to overcome resistance to AAV transduction, especially in hematopoietic cancer cells, whereas normal peripheral blood mononuclear cells and CD34(+) hematopoietic progenitor cells were not transduced. CONCLUSIONS: Consequently, recombinant targeted NQVGSWS AAV vectors harboring a suicide gene conferred selective killing to Kasumi-1 cells, but not to control cells. This suggests that the AAV mutant selected here may be used as a tool to target therapeutic genes to AML cells. PMID- 17920759 TI - Constitutive association of MyD88 to IRAK in HTLV-I-transformed T cells. AB - OBJECTIVE: Constitutive activation of nuclear factor (NF)-kappaB is a common feature of human T-cell leukemia virus type I (HTLV-I)-transformed T cells. Inhibition of NF-kappaB activity reduces cell growth and induces apoptosis of HTLV-I-transformed T cells, suggesting a central role of NF-kappaB in their proliferation and survival. In this study, we investigated whether MyD88, an adaptor protein of Toll-like receptor (TLR) signaling, contributes to constitutive NF-kappaB activation in HTLV-I-transformed T cells. MATERIALS AND METHODS: Activation status of MyD88 and interleukin (IL)-1R-associated kinase 1 (IRAK1) in HTLV-I-transformed human T cells, MT2, MT4, and HUT102 was examined by using Western blot and immunoprecipitation. TLR expression was evaluated with reverse transcription polymerase chain reaction. An expression vector encoding a dominant negative MyD88 with a deletion of its death domain (MyD88dn) was transfected into MT2 cells to evaluate roles of MyD88 in spontaneous activation of cytokine gene promoters and transcription factors, proliferation, and apoptosis in HTLV-I-transformed T cells. RESULTS: Constitutive association of MyD88 with IRAK1 was observed in all three of HTLV-I-transformed T cells, but not in HTLV-I-negative T cells, such as Jurkat, HUT78, and MOLT4. MT2 cells showed expression of TLR-1, -6, and -10 mRNAs. Constitutive activation of NF-kappaB and NF-IL-6 and cytokine gene promoters, such as IL-1alpha, interferon-gamma, and tumor necrosis factor-alpha in MT2 cells was inhibited by MyD88dn expression. MyD88dn reduced proliferation and induced apoptosis of MT2 cells. HTLV-I Tax enhanced TLR expression and synergistically activated NF-kappaB with wild-type MyD88. CONCLUSION: Our results show a novel pathway in NF-kappaB activation in HTLV-I-transformed T cells and further demonstrate a critical role of MyD88 in their dysregulated gene activation, survival, and proliferation. PMID- 17920760 TI - Detection of cryptic chromosomal lesions including acquired segmental uniparental disomy in advanced and low-risk myelodysplastic syndromes. AB - OBJECTIVES: Using metaphase cytogenetics (MC), chromosomal defects can be detected in 40% to 60% of patients with myelodysplastic syndromes (MDS); cytogenetic results have a major impact on prognosis. We hypothesize that more precise methods of chromosomal analysis will detect new/additional cryptic lesions in a higher proportion of MDS patients. METHODS: We have applied single nucleotide polymorphism microarrays (SNP-A) to perform high-resolution karyotyping in MDS to determine gene copy number and detect loss of heterozygosity (LOH). RESULTS: Using this method, chromosomal defects were found in 82% of MDS patients vs 50% as measured by MC; lesions were present in 68% of patients with normal MC, while in 81% of those with abnormal MC, new aberrations were found. In addition to gains or losses of chromosomal material, areas of LOH due to segmental uniparental disomy were found in 33% of patients. CONCLUSION: SNP-A findings demonstrate that chromosomal lesions are present in a much higher proportion of patients than predicted by traditional cytogenetics. These lesions may reflect an underlying generalized chromosomal instability in MDS. Additional previously cryptic defects may explain the clinical variability of MDS. New lesions may have important prognostic implications, suggesting that, in the future, SNP-A-based karyotyping may complement MC in laboratory evaluation of MDS. PMID- 17920761 TI - A new deal or a continuing dialogue of the deaf? PMID- 17920762 TI - In vitro and in vivo antitumor effects of doxorubicin loaded with bacterial magnetosomes (DBMs) on H22 cells: the magnetic bio-nanoparticles as drug carriers. AB - Hepatocellular carcinoma (HCC) is the most common form of cancer although effective therapeutic strategy especially targeted therapy is lacking. We recently employed bacterial magnetosomes (BMs) as the magnetic-targeted drug carrier and found an antitumor effect of doxorubicin (DOX)-loaded BMs (DBMs) in EMT-6 and HL60 cell lines. The aim of this study was to evaluate the in vitro and in vivo anti-neoplastic effects of DBMs on hepatic cancer. DBMs, DOX and BMs displayed tumor suppression rates of 86.8%, 78.6% and 4.3%, respectively, in H22 cell-bearing mice. The mortality rates following administration of DBMs, DOX and BMs were 20%, 80% and 0%, respectively. Pathological examination of hearts and tumors revealed that both DBMs and DOX effectively inhibited tumor growth although DBMs displayed a much lower cardiac toxicity compared with DOX. The DBMs were cytotoxic to H22 cells manifested as inhibition of cell proliferation and c myc expression, consistent with DOX. The IC(50) of DOX, DBMs and BMs in target cells were 5.309 +/- 0.010, 4.652 +/- 0.256 and 22.106 +/- 3.330 microg/ml, respectively. Our data revealed both in vitro and in vivo antitumor property of DBMs similar to that of DOX. More importantly, the adverse cardiac toxicity was significantly reduced in DBMs compared with DOX. Collectively, our study suggests the therapeutic potential of DBMs in target-therapy against liver cancer. PMID- 17920763 TI - Inhibition of Stat3 increases doxorubicin sensitivity in a human metastatic breast cancer cell line. AB - Metastatic breast cancer is an incurable disease, often characterized by poor response to standard chemotherapy, which is mainly based on anthracyclines and taxanes. Thus, increasing tumor cell sensitivity to these agents is an attractive goal towards improving the clinical management of this disease. The present study investigates the effects of signal transducer and activator of transcription 3 (Stat3) inhibition on the response of the highly metastatic MDA-MB-231 human breast adenocarcinoma cell line to doxorubicin (DOX). Stat3 is a transcription factor often constitutively activated in breast tumors and cancer cell lines, and is thought to contribute to malignant transformation and progression by transactivation of a host of target genes involved in cell proliferation and survival, angiogenesis and invasiveness. Our results indicate that (a) untreated MDA-MB-231 cells express higher baseline levels of (activated) pTyr(705)Stat3, that are further upregulated following exposure to DOX, than the non-metastatic MCF-7 cell line; (b) inhibiting the Stat3 signaling pathway, by exposure to the tyrphostin AG490 (an inhibitor of the upstream activating Janus kinases), by transfection with a dominant-negative form of Stat3 or by treatment with satraplatin (a tetravalent platinum derivative that inhibits Stat3 activation), increases breast cancer cell response to the proapoptotic effect of DOX (to different extents). In addition, the latter two approaches have been shown to interfere with expression of one or more antiapoptotic proteins. Overall, these observations suggest that suppression of Stat3 signaling may provide a potential therapeutic approach to overcoming DOX resistance in metastatic breast cancer cells. PMID- 17920765 TI - Decolorization of methylene blue in aqueous suspensions of titanium peroxide. AB - The pretreatment of TiO(2)-photocatalysts in solutions of H(2)O(2) was studied by examining the decolorization of methylene blue in the dark. Incubation of TiO(2) particles in H(2)O(2) solutions increased the oxidizing capacity of TiO(2). Methylene blue (0.3 mM) was degraded in the presence of pretreated TiO(2), and a decolorizing ratio of 47% was obtained after a 48-h incubation period in the presence of 5.0 g/L pretreated TiO(2). Titanium peroxide as a stable oxidant, which can be synthesized with the reaction of titanium sulfate and H(2)O(2), was studied in the decolorizing process of methylene blue. Concentrations of methylene blue were significantly reduced in the presence of titanium peroxide, and a greater extent of decolorization was obtained with larger amounts of titanium peroxide. A 63% decrease in methylene blue concentration was achieved in 5h incubation in the presence of 4.0 g/L titanium peroxide. H(2)O(2) accelerated the decolorizing process in the presence of titanium peroxide. The addition of 100 mM H(2)O(2) to a methylene blue solution containing 2.0 g/L titanium peroxide increased the decolorizing ratio to 85% after 5 h incubation. The addition of a hydroxyl radical scavenger, dimethyl sulfoxide, significantly decreased the decolorizing ratio, indicating the role of hydroxyl radicals in the oxidation process. PMID- 17920764 TI - Gambogic acid inhibits angiogenesis through suppressing vascular endothelial growth factor-induced tyrosine phosphorylation of KDR/Flk-1. AB - Previous studies revealed that gambogic acid (GA), the major active ingredient of gamboge, a brownish to orange resin exuded from Garcinia hanburryi tree in Southeast Asia, possessed significant anticancer activity both in vitro and in vivo. In this study, we explored the high antiangiogenic activities of GA for the first time. GA inhibits the VEGF-stimulated proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs) as well as microvessel sprouting from rat aortic rings in vitro. Moreover, GA inhibits vessel growth in matrigel plugs and CAM in vivo and transplanted tumor in mice. The results also indicated that GA decreases VEGF production of cultured tumor cells and inhibits VEGF-induced tyrosine phosphorylation of KDR/Flk-1. This inhibition of receptor phosphorylation is correlated with a significant decrease in VEGF-triggered phosphorylated forms of ERK, AKT and p38. Taken together, these findings strongly suggest that GA might be a structurally novel angiogenesis inhibitor. PMID- 17920767 TI - Continuous treatment process of mercury removal from aqueous solution by growing recombinant E. coli cells and modeling study. AB - A continuous treatment process was developed to investigate the capability of genetically engineered E. coli to simultaneously accumulate mercuric ions and reproduce itself in a continuous stirred tank reactor (CSTR) system. The influence of dilution rate and initial Hg(2+) concentration on continuous process was evaluated. Results indicated that the recombinant E. coli could effectively accumulate Hg(2+) from aqueous solution with Hg(2+) removal ratio up to about 90%, and propagate its cells at the same time in the continuous treatment system under suitable operational conditions. A kinetic model based on mass balance of Hg(2+) was proposed to simulate the continuous process. The modeling results were in good agreement with the experimental data. PMID- 17920766 TI - Experimental and mechanism research of SO(2) removal by cast iron scraps in a magnetically fixed bed. AB - Sulfur dioxide can be effectively removed by cast iron scraps corrosion process in a DC magnetically fixed bed, and iron sulfate compounds are gained as an available byproduct. At approximate 50 degrees C, when magnetic field intensity was at 15 mT and height of scraps was about 25 mm, the SO(2) removal efficiency can be kept above 80%. As the deposited rusts accumulated, the corrosion rate and desulfurization efficiency gradually decreased. The results show SO(2) removal efficiency depends on corrosion rate, and it can be obviously enhanced by DC magnetic field. With the XRD and SEM research, it can be found that DC magnetic field cannot change the crystal structure of rusts, but can make the surface morphologies on the surface of scraps looser which means easily to be removed. Consequently, the corrosion resistance can be lessened and SO(2) removal efficiency is improved significantly. PMID- 17920768 TI - The -181 A/C polymorphism in the excitatory amino acid transporter-2 gene promoter affects the personality trait of reward dependence in healthy subjects. AB - There have been some animal and human data suggesting that excitatory amino acid transporter (EAAT)-2, the major subtype of EAAT, is involved in human mental function and behavior. Recently, it has been shown that the -181 A/C polymorphism in the EAAT2 gene promoter affects plasma glutamate concentrations in humans. In the present study, the association of this genetic polymorphism with personality traits was examined in 575 Japanese healthy volunteers. Personality traits were assessed by the Temperament and Character Inventory, and the EAAT2 polymorphism was detected by a PCR-RFLP method. The scores of reward dependence were significantly (p=0.017) lower in the group with the A allele (A/A and A/C) than in that without this allele (C/C). When males and females were analyzed separately, the significant difference between the two genotype groups was observed in females (p=0.021) but not in males. The present study thus suggests that the -181 A/C polymorphism in the EAAT2 gene promoter affects the personality trait of reward dependence in healthy subjects. PMID- 17920769 TI - Bimodal effect of amphetamine on motor behaviors in C57BL/6 mice. AB - Amphetamine-induced motor behaviors, i.e., locomotor and stereotypic activities, were simultaneously characterized in C57BL/6 mice, a strain commonly used for genetic studies. Our findings show relatively high levels of focused activities in drug-naive C57BL/6 mice, confirming the lively nature of this mouse strain. Acute amphetamine induced a dose-dependent, bimodal response: locomotion predominated at lower doses of amphetamine and was gradually displaced by stereotypic behavior as dose and time increased. The sum total of both behavioral activities increased with amphetamine dose, supporting the notion that amphetamine-induced locomotion and stereotypy form a continuum. These data provide a basis for using C57BL/6 mice as a strain to study the molecular and cellular mechanisms underlying psychostimulant effects, drug addiction and psychotic disorders. PMID- 17920770 TI - Pharmacologic management of neuropathic pain: evidence-based recommendations. AB - Patients with neuropathic pain (NP) are challenging to manage and evidence-based clinical recommendations for pharmacologic management are needed. Systematic literature reviews, randomized clinical trials, and existing guidelines were evaluated at a consensus meeting. Medications were considered for recommendation if their efficacy was supported by at least one methodologically-sound, randomized clinical trial (RCT) demonstrating superiority to placebo or a relevant comparison treatment. Recommendations were based on the amount and consistency of evidence, degree of efficacy, safety, and clinical experience of the authors. Available RCTs typically evaluated chronic NP of moderate to severe intensity. Recommended first-line treatments include certain antidepressants (i.e., tricyclic antidepressants and dual reuptake inhibitors of both serotonin and norepinephrine), calcium channel alpha2-delta ligands (i.e., gabapentin and pregabalin), and topical lidocaine. Opioid analgesics and tramadol are recommended as generally second-line treatments that can be considered for first line use in select clinical circumstances. Other medications that would generally be used as third-line treatments but that could also be used as second-line treatments in some circumstances include certain antiepileptic and antidepressant medications, mexiletine, N-methyl-D-aspartate receptor antagonists, and topical capsaicin. Medication selection should be individualized, considering side effects, potential beneficial or deleterious effects on comorbidities, and whether prompt onset of pain relief is necessary. To date, no medications have demonstrated efficacy in lumbosacral radiculopathy, which is probably the most common type of NP. Long-term studies, head-to-head comparisons between medications, studies involving combinations of medications, and RCTs examining treatment of central NP are lacking and should be a priority for future research. PMID- 17920771 TI - Pain in patients with cancer: still a long way to go. PMID- 17920772 TI - Endocytosis of pulchellin and its recombinant B-chain into K-562 cells: binding and uptake studies. AB - Most of the type 2 ribosome-inactivating proteins (RIPs) are toxins formed by an RNA-N-glycosidase A-chain polypeptide linked to a lectin B-chain by a single disulfide bond. Members of this protein class vary greatly in cytotoxity, correlating more with B-chain diversity rather than to A-chain differences. Pulchellin is a type 2 ribosome-inactivating protein toxin found in the seeds of Abrus pulchellus tenuiflorus. Recombinant pulchellin B-Chain (rPBC) has been previously produced as inclusion bodies in Escherichia coli and successfully refolded recovering biological activity. New approaches for using this kind of protein as a biotechnological tool require a better understanding of cell targeting, binding, uptake, intracellular routing and delivery. In this work, cell adhesion experiments were used to determine the interaction of rPBC with mammalian cells. Fluorescence and confocal microscopy revealed the intracellular localization and trafficking. Subcellular sorting of the native pulchellin could also be determined. The results support that the endosomal internalization pathway and the retrograde transport through the Golgi apparatus might be used by both native protein and rPBC. PMID- 17920773 TI - Antibody-based targeting of the tumor vasculature. AB - Conventional cytotoxic therapies of cancer often suffer from a lack of specificity, leading to a poor therapeutic index and considerable toxicities to normal organs. An elegant way to overcome the disadvantages of conventional tumor therapy is the selective delivery of therapeutics to the tumor site by their conjugation to a carrier molecule specific for a tumor-associated molecular marker. Markers expressed on the tumor's vasculature represent particularly attractive targets for a site-specific pharmacodelivery due to their inherent accessibility for blood-borne agents and the various therapeutic options that they allow, ranging from intraluminal blood coagulation to the recruitment of immune cells. In this review, we will outline advances in the preclinical and clinical evaluation of antibody-based vascular targeting agents, describe technologies for the discovery of novel vascular targets and discuss future prospects for vascular targeting applications. PMID- 17920774 TI - Assessment of burn area: most objective method. PMID- 17920775 TI - Loss of capsaicin-induced meningeal neurogenic sensory vasodilatation in diabetic rats. AB - Neuropathic alterations of sensory nerves involved in the mediation of neurogenic inflammation of the meninges may contribute to the increased incidence of headaches in diabetics. In the rat, activation of capsaicin-sensitive nociceptors, which express the transient receptor potential vanilloid type 1 (TRPV1) receptor, induces meningeal vasodilatation, a significant component of neurogenic inflammation, through the release of calcitonin gene-related peptide (CGRP). This study examines the effects of streptozotocin-induced diabetes on TRPV1 receptor-mediated neurogenic sensory vasodilatation, CGRP release and nerve fiber density in the rat dura mater. In a cranial window preparation, epidural application of capsaicin (10(-7) M) produced distinct vasodilatory responses in control animals as measured by laser Doppler flowmetry. In diabetic rats, capsaicin-induced vasodilatation was reduced or even abolished 6, but not 2 or 4 weeks after diabetes induction. In contrast, vasoconstriction, a non-neurogenic response to capsaicin at a higher concentration (10(-5) M), was not altered in diabetic rats. The vasodilatory effects of histamine (10(-5) M), acetylcholine (10(-4) M) and CGRP (10(-5) M) were similar in control, diabetic and insulin treated diabetic animals. In diabetic rats, a significant decrease in the capsaicin-evoked release of CGRP and reduction in the density of TRPV1 immunoreactive (IR) nerves were demonstrated. Treatment of the diabetic rats with insulin restored both the vasodilatory response and the capsaicin-induced CGRP release toward control values. In conclusion, this study revealed a marked impairment of meningeal TRPV1-IR nerves in streptozotocin diabetic rats by showing reduced neurogenic sensory vasodilatation, decreased capsaicin-evoked CGRP release and reduction in the number of TRPV1-IR nerve fibers of the dura mater. The findings suggest that capsaicin-sensitive afferents may play an important role in meningeal nociceptor function and their dysfunction, e.g. due to a limited removal of inflammatory mediators and/or tissue metabolites from the meningeal tissue, may contribute to the enhanced incidence of headaches in diabetics. PMID- 17920777 TI - The burden of untreated ADHD among adults: the role of stimulant medication. AB - The present study investigated to what degree medication with stimulants for children with ADHD will reduce comorbid complications in adulthood. Two groups (N=17 and N=74) were defined retrospectively using data from archives at the Expert team for ADHD for Middle and Northern Norway. The groups were compared on the "Index of Burden", an index constructed on the basis of data from five variables; Alcohol abuse, substance abuse, criminality, the Global Severity Index of SCL-90 and the "Functioning and quality of life" questionnaire. Results showed that treatment with stimulants in childhood/youth contribute to increased social and psychological functioning in adult age. PMID- 17920776 TI - Education modulates cortisol reactivity to the Trier Social Stress Test in middle aged adults. AB - The present study assessed the modulating effect of education level on cortisol reactivity to the Trier Social Stress Test (TSST) in a sample of 101 middle-aged adults (22 males, 79 females) between the ages of 50 and 65. The TSST involves a public speech and mental arithmetic task in front of an audience. No previous studies have assessed whether education level can have an impact on cortisol reactivity to this psychosocial stressor. It is plausible that greater exposure to academia may impact how one perceives and responds to the demands of the speech and arithmetic task. Should education have an impact on cortisol reactivity to the TSST, future studies will be required to control for this factor in order to reduce both statistical error and false interpretations. In addition to completing the TSST, participants were administered a battery of neurocognitive tests and personality questionnaires, including a report on education level (i.e. number of years total and degree: High School, Junior College, Technical, University). Results showed that adults with post-secondary education above Junior College tended to secrete higher cortisol levels overall, as measured by total area under the curve. However, it was the group with lower educational attainment who showed a greater stress response specific to the TSST, as measured by percentage increase in cortisol from pre- to post-TSST. Analyses also found that higher educated adults performed better than their less educated peers on verbal fluency. Considering that the TSST is an oral task, it is suggested that middle-aged individuals with a lower level of education may find the TSST more stressful due to lower verbal capacity, which may lead to an increased cortisol response to the TSST when compared to individuals with a higher level of education. PMID- 17920778 TI - Random number generation and creativity. AB - A previous paper suggested that humans can generate genuinely random numbers. I tested this hypothesis by repeating the experiment with a larger number of highly numerate subjects, asking them to call out a sequence of digits selected from 0 through 9. The resulting sequences were substantially non-random, with an excess of sequential pairs of numbers and a deficit of repeats of the same number, in line with previous literature. However, the previous literature suggests that humans generate random numbers with substantial conscious effort, and distractions which reduce that effort reduce the randomness of the numbers. I reduced my subjects' concentration by asking them to call out in another language, and with alcohol - neither affected the randomness of their responses. This suggests that the ability to generate random numbers is a 'basic' function of the human mind, even if those numbers are not mathematically 'random'. I hypothesise that there is a 'creativity' mechanism, while not truly random, provides novelty as part of the mind's defence against closed programming loops, and that testing for the effects seen here in people more or less familiar with numbers or with spontaneous creativity could identify more features of this process. It is possible that training to perform better at simple random generation tasks could help to increase creativity, through training people to reduce the conscious mind's suppression of the 'spontaneous', creative response to new questions. PMID- 17920779 TI - 2-deoxy-D-glucose as a potential drug against fusogenic viruses including HIV. AB - Most enveloped viruses fuse with host cells and catalyze fusion among host cells by expression of specific patterns of N-glycosylation on their envelope proteins. In the 1970s, it was observed that 2-deoxy-D-glucose (2DOG) and 2-fluoro-2-deoxy D-mannose (2F2DOM) inhibited N-glycosylation of asparagine (Asn) sites on the external domain of viral envelope proteins. This effect led to the virus particles being non-fusogenic with greatly reduced infectivity and reduced ability to pass from cell to cell by catalyzing cell--cell fusion. At that time, this observation was not particularly important because viral diseases were readily prevented by vaccines and there was no known link between fusogenic viruses and cancer. Today, we are faced with a chronic and lethal viral disease (AIDS) caused by a virus (HIV) that mutates so quickly that we have not been able to produce a vaccine. Moreover, it is spreading among millions of people unable to afford more than basic medications. In addition, cell--cell fusion has been identified as an important, if not essential, step in the progression of abnormal cell clones to clinically significant cancer and fusogenic viruses have been shown to cause progression of some tumors. Here, we reiterate the hypothesize (first made in 1986 by Blough et al. [Blough HA, Pauwels R, De Clercq E, Cogniaux J, Sprecher-Goldberger S, Thiry L. Glycosylation inhibitors block the expression of LAV/HTLV-III (HIV) glycoproteins. Biochem Biophys Res Commun 1986; 141:33-8]) that 2DOG, 2F2DOM and related compounds, which interfere with normal N glycosylation of virus envelope proteins, are attractive candidates for anti fusogenic drugs that can be used against chronic virus diseases and cancers. This analysis also supports the concept of blocking N-acetylglucosaminyl-transferases with chloroquine or other drugs (proposed by Savarino et al. [Savarino A, Lucia MB, Rastrelli E et al. Anti-HIV effects of chloroquine: inhibition of viral particle glycosylation and synergism with protease inhibitors. J Acquir Immune Defic Syndr 2004; 35:223-32]) as an anti-viral approach. These drugs may have broader utility and lower cost than drugs designed specifically to target the gp41 protein of HIV, which have become popular as viral-entry inhibitors for treatment of HIV/AIDS. PMID- 17920780 TI - A novel hypothesis of atherosclerosis: EPCs-mediated repair-to-injury. AB - Recent findings demonstrate the vital role of endothelial progenitor cells in the homeostasis of the vessel wall and the development of atherosclerosis. Endothelial progenitor cells (EPCs) play important roles in repair-to-injury of arteries. Many evidences have shown Cardiovascular risk factors closely correlated with EPCs numbers and function. Levels of circulating EPCs represented a better predictor of endothelial function than conventional risk factors. Depletion of bone marrow and Cardiovascular risk factors are the two prerequisits of atherosclerosis. All conditions of manifest atherosclerotic disease are accompanied by reduced EPC numbers and migratory capacity. Therefore, based on response-to-injury hypothesis and these findings, we build up EPCs-mediated repair-to-injury hypothesis, which may have important therapeutic implications in the prevention and therapy of atherosclerosis. The use of EPCs for vascular repair may be important therapy strategies with a maximized benefit for the patient in the future. PMID- 17920781 TI - Segmental arterial mediolysis: unrecognized cases culled from cases of ruptured aneurysm of abdominal visceral arteries reported in the Japanese literature. AB - Segmental arterial mediolysis (SAM) is a rare nonatherosclerotic and noninflammatory arteriopathy that was proposed by Slavin et al. [Segmental mediolytic arteritis. A clinical pathologic study, Lab. Invest. 35 (1976) 23-29]. It mainly involves abdominal visceral arteries and is characterized by lytic degeneration of the media, resulting in intraabdominal bleeding. We collected 27 unrecognized cases of SAM by reviewing microscopic slides of cases of ruptured aneurysms of visceral arteries, except splenic and hepatic aneurysms, reported in the Japanese literature. This paper describes the pathological and clinical features of these cases. The symptom at onset was abdominal pain associated with intraabdominal bleeding in all cases. The most involved artery was the middle colic artery, accounting for 14 (50%), followed by gastric and gastroepiploic arteries, (6 and 5, respectively). Seventy-eight percent of aneurysms were of dissecting type and the rest of pseudoaneurysm type, except for one. Multiple aneurysms were found in 9 cases (33.3%). Pathological lesions were acute in all. The outcome of those who had surgery was good, even in those who had surgery for 1 ruptured aneurysm, leaving the others unmanaged. The relationship of SAM to fibromuscular dysplasia is discussed. Secondary changes in the wall of the accompanying vein to the affected artery are briefly described. It is emphasized that the majority of aneurysms of abdominal visceral arteries are gathered together as SAM as a definite clinical and pathological entity. PMID- 17920782 TI - An approach for practical multiobjective IMRT treatment planning. AB - PURPOSE: To introduce and demonstrate a practical multiobjective treatment planning procedure for intensity-modulated radiation therapy (IMRT) planning. METHODS AND MATERIALS: The creation of a database of Pareto optimal treatment plans proceeds in two steps. The first step solves an optimization problem that finds a single treatment plan which is close to a set of clinical aspirations. This plan provides an example of what is feasible, and is then used to determine mutually satisfiable hard constraints for the subsequent generation of the plan database. All optimizations are done using linear programming. RESULTS: The two step procedure is applied to a brain, a prostate, and a lung case. The plan databases created allow for the selection of a final treatment plan based on the observed tradeoffs between the various organs involved. CONCLUSIONS: The proposed method reduces the human iteration time common in IMRT treatment planning. Additionally, the database of plans, when properly viewed, allows the decision maker to make an informed final plan selection. PMID- 17920783 TI - Quantification of mediastinal and hilar lymph node movement using four dimensional computed tomography scan: implications for radiation treatment planning. AB - PURPOSE: To quantitatively describe mediastinal and hilar lymph node movement in patients with lymph node-positive lung cancer. METHODS AND MATERIALS: Twenty-four patients with lung cancer who underwent four-dimensional computed tomography scanning at Massachusetts General Hospital were included in the study. The maximum extent of superior motion of the superior border was measured, as well as the maximum inferior movement of the inferior border. The average of these two values is defined as the peak-to-peak movement. This process was repeated for mediolateral (ML) and anterior-posterior (AP) movement. Linear regression was used to determine lymph node characteristics associated with peak-to-peak movement. Various uniform expansions were investigated to determine the expansion margins necessary to ensure complete internal target volume (ITV) coverage. RESULTS: The mean peak-to-peak displacements of paratracheal lymph nodes were 4 mm (craniocaudal [CC]), 2 mm (ML), and 2 mm (AP). For subcarinal lymph nodes, the mean peak-to-peak movements were 6 mm (CC), 4 mm (ML), and 2 mm (AP). The mean peak-to-peak displacements of hilar lymph nodes were 7 mm (CC), 1 mm (ML), and 4 mm (AP). On multivariate analysis, lymph node station and lymph node size were significantly related to peak-to-peak movement. Expansions of 8 mm for paratracheal nodes and 13 mm for subcarinal and hilar nodes would have been necessary to cover the ITV of 95% of these nodal masses. CONCLUSIONS: Subcarinal and hilar lymph nodes may move substantially throughout the respiratory cycle. In the absence of patient-specific information on nodal motion, expansions of at least 8 mm, 13 mm, and 13 mm should be considered to cover the ITV of paratracheal, subcarinal, and hilar lymph nodes, respectively. PMID- 17920785 TI - [Compliance to the antibiotic committee guidelines in Tourcoing Hospital]. AB - AIMS OF THE STUDY: We evaluated the compliance to the antibiotic committee guidelines of Tourcoing Hospital. METHODS: A national nosocomial infections prevalence study was conducted in June 2006. We collected additional data on the name and indication of prescribed antibiotics compared to recommended drugs in our therapeutic guidelines. Endpoints were antibiotic indication, compliance to local guidelines, unjustified combination therapy and deescalation therapy if possible. Situations non included in the guidelines were evaluated on a case to case basis after discussion with the prescribing physician. Pediatric (N=5) or prophylaxis (N=4) prescriptions were not analysed. RESULTS: Antibiotics were used in 97/669 (14.5%) patients including 32% in acute care, 11% in rehab and 0,9% in long term care. Drugs recommended in the guidelines were used in 63 cases (60.5%) including 56.3% first line and 4.2% justified second line therapy. When including situations not included in the guidelines and judged as correct, compliance reached 64.9%. The 41 variations from guidelines observed in 34 patients concerned: molecule choice (N=12), lack of antibiotic indication (N=12), unjustified combination therapy (N=12), drug choice in combination therapy (N=5), lack of deescalation (N=1). Lower respiratory tract (N=12) and urinary tract (N=7) infections as well as fluoroquinolones (N=12) were the most frequent deviations from guidelines. CONCLUSION: Compliance rate is encouraging. This study pinpoints specific targets for future interventions. PMID- 17920784 TI - Radiation effects on the cytoskeleton of endothelial cells and endothelial monolayer permeability. AB - PURPOSE: To investigate the effects of radiation on the endothelial cytoskeleton and endothelial monolayer permeability and to evaluate associated signaling pathways, which could reveal potential mechanisms of known vascular effects of radiation. METHODS AND MATERIALS: Cultured endothelial cells were X-ray irradiated, and actin filaments, microtubules, intermediate filaments, and vascular endothelial (VE)-cadherin junctions were examined by immunofluorescence. Permeability was determined by the passage of fluorescent dextran through cell monolayers. Signal transduction pathways were analyzed using RhoA, Rho kinase, and stress-activated protein kinase-p38 (SAPK2/p38) inhibitors by guanosine triphosphate-RhoA activation assay and transfection with RhoAT19N. The levels of junction protein expression and phosphorylation of myosin light chain and SAPK2/p38 were assessed by Western blotting. The radiation effects on cell death were verified by clonogenic assays. RESULTS: Radiation induced rapid and persistent actin stress fiber formation and redistribution of VE-cadherin junctions in microvascular, but not umbilical vein endothelial cells, and microtubules and intermediate filaments remained unaffected. Radiation also caused a rapid and persistent increase in microvascular permeability. RhoA guanosine triphosphatase and Rho kinase were activated by radiation and caused phosphorylation of downstream myosin light chain and the observed cytoskeletal and permeability changes. SAPK2/p38 was activated by radiation but did not influence either the cytoskeleton or permeability. CONCLUSION: This study is the first to show rapid activation of the RhoA/Rho kinase by radiation in endothelial cells and has demonstrated a link between this pathway and cytoskeletal remodeling and permeability. The results also suggest that the RhoA pathway might be a useful target for modulating the permeability and other effects of radiation for therapeutic gain. PMID- 17920786 TI - [Genotyping diagnosis of acyclovir resistant herpes simplex virus]. AB - Herpes simplex virus resistant to acyclovir (ACV) is a major concern among immunocompromised patients. ACV resistance might be due to mutations located in one of the two genes involved in ACV mechanism of action, the thymidine kinase gene (TK, involved in 95% of the cases) and the DNA polymerase gene. TK gene mutations consist, in half of the cases, in nucleotide insertion or deletion, occurring most of the time in G or C homopolymers considered as hot spots. Half of the other cases involves nucleotide substitutions leading to amino acids substitutions. Studies of sensitive strains revealed a high degree of TK polymorphism, many mutations being not implied in ACV resistance. At the present time, resistance detection can be performed by phenotypic tests that require virus culture and results cannot be given to the physician before 7 to 10 days. Genotyping diagnosis performed directly from clinical samples would allow to detect resistance more rapidly, in order to switch quickly to an appropriate treatment by foscarnet or cidofovir. PMID- 17920788 TI - Isolation of the three grape sub-lineages of B-class MADS-box TM6, PISTILLATA and APETALA3 genes which are differentially expressed during flower and fruit development. AB - The B class of MADS-box floral homeotic genes specifies petal and stamen identity in angiosperms. While this group is one of the most studied in herbaceous plant species, it has remained largely uncharacterized in woody species such as grapevine. Although the B class PI/GLO and AP3/DEF clades have been extensively characterized in model species, the role of the TM6 subgroup within the AP3 clade is not completely understood, since it is absent in Arabidopsis thaliana. In this study, the coding regions of VvTM6 and VvAP3 and the genomic sequence of VvPI, were cloned. VvPI and AtPI were confirmed to be functional homologues by means of complementation of the pi Arabidopsis mutant. Expression analysis revealed that VvPI and VvAP3 transcripts are restricted almost exclusively to inflorescences, although VvPI was detected at low levels in leaves and roots. VvTM6 expresses throughout the plant, with higher levels in flowers and berries. A detailed chronological study of grape flower progression by light microscopy and temporal expression analysis throughout early and late developmental stages, revealed that VvPI expression increases during pollen maturation and decreases between the events of pollination and fertilization, before the cap fall. On the other hand, VvTM6 is expressed in the last stage of anther development. Specific expression of VvAP3 and VvPI was detected in petals and stamens within the flower, while VvTM6 was also expressed in carpels. Moreover, this work provides the first evidence for expression of a TM6-like gene throughout fruit growth and ripening. Even if these genes belong to the same genetic class they could act in different periods and/or tissues during reproductive organ development. PMID- 17920789 TI - Substitution of synthetic chimpanzee androgen receptor for human androgen receptor in competitive binding and transcriptional activation assays for EDC screening. AB - The potential effect of receptor-mediated endocrine modulators across species is of increasing concern. In attempts to address these concerns, we are developing androgen and estrogen receptor binding assays using recombinant hormone receptors from a number of species across different vertebrate classes. The United States Environmental Protection Agency (USEPA) Office of Science Coordination and Policy (OSCP) requested that we develop a nonhuman mammalian receptor-binding assay for possible use in their Endocrine Disruptor Screening Program (EDSP). Since the chimpanzee androgen receptor is very similar to that of humans and thus possesses properties which could be exploited in future endocrine studies, we synthesized and expressed this gene in eukaryotic expression plasmids, baculovirus expression vectors and replication deficient adenovirus. In all ligand-binding and transcriptional activation assays tested, the chimpanzee receptor performed essentially identically to the human receptor. This suggests that the chimpanzee gene could substitute for the human gene in endocrine screening assays. PMID- 17920787 TI - Ketamine exposure in adult mice leads to increased cell death in C3H, DBA2 and FVB inbred mouse strains. AB - BACKGROUND: Drug abuse is common among adolescents and young adults. Although the consequences of intoxication are known, sequelae of drugs emerging on campuses and in clubs nationwide are not. We previously demonstrated that ketamine exposure results in lasting physiological abnormalities in mice. However, the extent to which these deficits reflect neuropathologic changes is not known. METHODS: The current study examines neuropathologic changes following sub anesthetic ketamine administration (5mg/kg i.p. x 5) to three inbred mouse strains. Stereologic quantification of silver stained nuclear and linear profiles as well as activated caspase-3 labeling was used to address: (1) whether or not ketamine increases excitotoxic and apoptotic cell death in hippocampal CA3 and (2) whether or not ketamine-induced cell death varies by genetic background. RESULTS: Ketamine increased cell death in hippocampal CA3 of adult C3H, DBA2 and FVB mice. Neither silver staining nor activated caspase-3 labeling varied by strain, nor was there an interaction between ketamine-induced cell death and strain. CONCLUSIONS: Ketamine exposure among young adults, even in limited amounts, may lead to irreversible changes in both brain function and structure. Loss of CA3 hippocampal cells may underlie persistent ERP changes previously shown in mice and possibly contribute to lasting cognitive deficits among ketamine abusers. PMID- 17920791 TI - A new approach to the toxicity testing of carbon-based nanomaterials--the clonogenic assay. AB - The cellular toxicity of three types of carbon nanoparticles, namely HiPco single walled carbon nanotubes (SWCNT), arc discharge SWCNT and Printex 90 carbon black nanoparticles, was studied on three different cell models including the human alveolar carcinoma epithelial cell line (A549), the normal human bronchial epithelial cell line (BEAS-2B) and the human keratinocyte cell line (HaCaT) using the clonogenic assay. Carbon nanomaterials are known to interact with colorimetric indicator dyes frequently used in cytotoxicity assays. By employing the clonogenic assay, any such interactions could be avoided, allowing a more reliable method for the in vitro toxicity assessment of carbon-based nanoparticles. It could be shown that the toxicity of as produced SWCNT samples differs between cell lines and the SWCNT production method used, with HiPco SWCNT samples being more reactive compared to arc discharge produced SWCNT samples, both eliciting a stronger cytotoxic response than carbon black. Furthermore, it was possible to distinguish between effects on cell viability and cell proliferation by including colony size as an additional endpoint in the clonogenic assay. All three particle types were highly effective in inhibiting cell proliferation in all three cell lines, whereas only HaCaT and BEAS-2B cells also showed decreased cell viability. PMID- 17920790 TI - Rapid effects of diesel exhaust particulate extracts on intracellular signaling in human endothelial cells. AB - Inhalation of ultrafine particulate matter (PM) in air pollution increases cardiovascular mortality by passing into systemic circulation and possibly affecting endothelial cell (EC) function. This study identified the chemical constituents, including polycyclic aromatic hydrocarbons (PAHs), in diesel exhaust particulate extracts (DEPEs) prepared from a truck run at different speeds and engine loads. The short-term effects of DEPEs alone or in combination with estradiol (E(2)) on MAPK (ERK1/2), AKT, and eNOS activation and nitric oxide (NO) production in human umbilical vein EC (HUVEC) were evaluated. Notably, DEPE from a truck run under increasing loads (L) stimulated phosphorylation of MAPK, AKT, and eNOS whereas DEPE from the truck run at increasing speeds (S) did not affect MAPK alone, but inhibited E(2)-induced MAPK and eNOS phosphorylation. Higher PAH concentrations in the DEPE L versus DEPE S samples correlate with the observed differences in cellular activities. Like E(2), DEPEs rapidly increased NO with the DEPE L sample acting additively with E(2) and then inhibiting E(2) induced NO with longer treatment time. Like E(2), DEPEs increased trans endothelial electrical resistance (TEER) across a monolayer of HUVEC. These data are the first characterization of rapid effects of DEPE in human EC and may indicate mechanisms for diesel exhaust in vascular function. PMID- 17920792 TI - Turkish women's level of knowledge on and attitude toward sexual health. AB - OBJECTIVE: Sexuality is a basic human function that can affect general well-being and overall life quality. The aim of this study was to assess the level of knowledge on and attitude toward sexuality of a subgroup of Turkish women. METHODS: A cross-sectional survey including 1007 women was carried out concurrently in gynecology and urology outpatient clinics of Gazi University Faculty of Medicine, Turkey, between January 2003 and December 2003. A self created questionnaire for data collection was administered to the participants, which addressed questions concerning their sexual life, sexual problems and attitudes towards sexuality. Statistical package for the social sciences (SPSS) was used for the data analysis of the study (SPSS ver. 11.0 Inc. Chicago, IL, USA). RESULTS: The data included 1007 patients aged 14-77 (mean: 35.6 +/- 11.1). Among the women assessed, 90.2% were married, 79.8% were premenopausal and 77.1% were currently sexually active. 68.7% entered sexual intercourse 1-2 times per week, while 28.5% had an average of 3-4. 77.8% were aware of experiencing an orgasm and 74.7% had at least one orgasm. Only 21.4% of all women had an orgasm during each sexual intercourse. 2.2% of those who had never experienced an orgasm concurrently applied to a medical center. The average age of first intercourse was 21.3 +/- 4.1 years. CONCLUSIONS: Because there is no national data on sexual health in Turkey, analysis and evaluation of the severity of the problem, and alternatives for international comparisons are not possible. In our opinion, this study presents a valuable perception into the current sexual behavior of Turkish women. PMID- 17920793 TI - Self-assembled drug delivery systems 2. Cholesteryl derivatives of antiviral nucleoside analogues: synthesis, properties and the vesicle formation. AB - Self-assembled drug delivery systems (SADDS) are defined as the self-aggregates of amphiphilic prodrugs. Prodrug, molecular self-assembly and nanotechnology are involved in SADDS manufacturing. But the knowledge of the self-assembly of amphiphilic prodrugs and the formation rules of SADDS is very limited. In this paper, five cholesteryl derivatives of antiviral nucleoside analogues were synthesized, involving antiviral acyclovir, didanosine and zidovudine, and the different acyl linkers, succinyl, adipoyl and phosphoryl. The derivatives are typical amphiphiles with nucleosides as polar heads and long-chained lipids as hydrophobic tails. The derivatives showed the similar soluble behavior, and the solubility highly depended on the types of solvents. Two forces, hydrogen bonding and hydrophobic interaction in alcohol solutions could improve the derivatives dissolving. However, the molecular self-assembly of derivatives could prefer to happen in the noncompetitive solvents including chloroform and tetrahydrofuran (THF) based on the intermolecular hydrogen bonding between nucleobase moieties, which could greatly increase their solubility. The derivatives formed nanosized vesicles based on hydrophobic interaction after injecting their THF solutions into water. The volume ratios of polar heads and hydrophobic tails of amphiphiles could determine the vesicle size, and the amphiphiles with large ratios would prefer to form small vesicles. The self-assembled vesicles would likely become SADDS. PMID- 17920794 TI - Preparation and characterization of intravenously injectable nimodipine nanosuspension. AB - The purpose of this study was to develop an alternative, improved and better tolerated injectable nimodipine nanosuspension compared with commercially available ethanol solution. In this study, nimodipine nanosuspension was prepared by high-pressure homogenization (HPH). The effects of the production parameters such as pressure, cycle numbers and crushing principles on the mean particle size, 99% diameter and polydispersity of the nanosuspension were investigated. Characterization of the product was performed by scanning electron microscope (SEM) and differential scanning calorimeter (DSC). The safety of the nimodipine nanosuspension was discussed with special attention to contamination by microparticles and the increase in saturation solubility C(s). Irritability study in rabbits showed that this formulation provided less local irritation and phlebitis risks than the commercial ethanol product, which represented a promising new drug formulation for intravenous therapy of subarachnoid hemorrhage (SAH)-related vasospasm. PMID- 17920795 TI - A novel ketoconazole bioadhesive effervescent tablet for vaginal delivery: design, in vitro and 'in vivo' evaluation. AB - Bioadhesive tablet formulations of ketoconazole for vaginal delivery were studied. Carbomer (Carbopol 974P, Carbopol 934P), hydroxypropylmethyl cellulose (HPMC) and hydroxypropyl cellulose (HPC) were used as candidate bioadhesive polymers. Effervescent was incorporated into the formulations as a disintegration agent. The swelling behavior and bioadhesive strength of the drug-free tablets were investigated. Carbopol 934P was selected as biopolymer in combination with HPMC or HPC at different ratios to develop five drug-loaded formulations. The swellings, tackiness and in vitro release were studied on the tablets. A good sustained effect and a moderate bioadhesion were obtained with the tablets. The formulation containing 100mg of effervescent, with the Carbopol 934P:HPC ratio of 1:9, seemed to be the optimum one for the tablet. In vivo drug residence tests were carried out by administering the preferred formulation to female rats. The results showed that the drug remaining followed a one-order model. Even after 24h of administration in vagina of rats, 17% of the original employed drug was retained on the vaginal tissue. Our study may provide a potential vaginal tablet formulation of ketoconazole against Candida albicans. PMID- 17920796 TI - Physicochemical properties of water and its effect on drug delivery. A commentary. AB - The structure and properties of water are integral to the existence and evolution of life on any number of levels. Consistent with this overarching statement, the unique physiochemical properties of water affect the pharmacological actions and delivery of drugs to the body whether they are administered orally, topically or by injection. This last topic is explored in the current review. While water is a group VIA hydride, it is distinct from other members of the class based on density, dielectric constant, surface tension as well as melting and boiling point. These differences are attributed to the ability of water to hydrogen bond to itself and other substrates resulting in the formation of strongly cohesive systems which molecularly resemble highly dynamic polymeric networks. As a consequence of these properties, hydrophobic compounds tend to aggregate in solution sometimes at the nanoscale. The practical consequence of this aggregation may be observed as spurious results associated with receptor-based high throughput screening assays as well as anomalies in phase-solubility analysis encountered in the study of hydrophobic materials with cyclodextrins. Other insights provided by a knowledge of the structure of water include the actions of excipients. Thus, materials that contribute to the hydrogen-bonding aqueous network (i.e., kosmotropes) will tend to salt more non-polar materials out of solution while material that destabilize the network structures (i.e., chaotropes) will tend to preferentially bind to solutes, reducing unfavorable interactions with water, resulting in solubilization. At membranes, the unique properties of water can affect drug absorption based on resistance in the unstirred water layer (UWL) which resides directly adjacent to the barrier. Depending on the nature of the membrane and the drug, the UWL can effectively reduce drug uptake and penetration. Furthermore, excipients that affect water structure can either contribute to or detract from the ability of a compound to pass the UWL and consequently the membrane. The increasing realization that water influences the actions and interactions drugs and excipients opens a variety of new avenues with regard to the rationale design of useful dosage forms. PMID- 17920797 TI - Strategies to regenerate hair cells: identification of progenitors and critical genes. AB - Deafness commonly results from a lesion of the sensory cells and/or of the neurons of the auditory part of the inner ear. There are currently no treatments designed to halt or reverse the progression of hearing loss. A key goal in developing therapy for sensorineural deafness is the identification of strategies to replace lost hair cells. In amphibians and birds, a spontaneous post-injury regeneration of all inner ear sensory hair cells occurs. In contrast, in the mammalian cochlea, hair cells are only produced during embryogenesis. Many studies have been carried out in order to demonstrate the persistence of endogenous progenitors. The present review is first focused on the occurrence of spontaneous supernumerary hair cells and on nestin positive precursors found in the organ of Corti. A second approach to regenerating hair cells would be to find genes essential for their differentiation. This review will also focus on critical genes for embryonic hair cell formation such as the cell cycle related proteins, the Atoh1 gene and the Notch signaling pathway. Understanding mechanisms that underlie hair cell production is an essential prerequisite to defining therapeutic strategies to regenerate hair cells in the mature inner ear. PMID- 17920798 TI - A place principle for vertigo. AB - OBJECTIVE: To provide a road map of the vestibular labyrinth and its innervation leading to a place principle for different forms of vertigo. METHOD: The literature describing the anatomy and physiology of the vestibular system was reviewed. RESULTS: Different forms of vertigo may be determined by the type of sense organ, type of ganglion cell and location in the vestibular nerve. CONCLUSION: Partial lesions (viral) of the vestibular ganglion are manifested as various forms of vertigo. PMID- 17920799 TI - Characterization of language and reading skills in familial polymicrogyria. AB - Polymicrogyria (PMG) is a malformation of cortical development characterized by an excessive number of small gyri and abnormal cortical lamination, giving the cortical surface an irregular and gross appearance. The severity of clinical manifestations correlates with the extent of cortical involvement. The objective of the present study was to describe three families with linguistic features of developmental language disorder and reading impairment, and to establish a neuroanatomic correlation through neuroimaging. Subjects have been submitted to a comprehensive protocol including psychological assessment, language evaluation, neurological examination, and neuroimaging investigation. In our families, children usually had the diagnosis of developmental language disorder while adults had the diagnosis of reading impairment. MRI showed perisylvian polymicrogyria in several subjects of each family. Our data support the idea that there is a co-occurrence of developmental language disorder and reading impairment and both conditions may be associated with polymicrogyria. PMID- 17920800 TI - Limited value of shape, margin and CT density in the discrimination between benign and malignant screen detected solid pulmonary nodules of the NELSON trial. AB - PURPOSE: To evaluate prospectively the value of size, shape, margin and density in discriminating between benign and malignant CT screen detected solid non calcified pulmonary nodules. MATERIAL AND METHODS: This study was institutional review board approved. For this study 405 participants of the NELSON lung cancer screening trial with 469 indeterminate or potentially malignant solid pulmonary nodules (>50mm3) were selected. The nodules were classified based on size, shape (round, polygonal, irregular) and margin (smooth, lobulated, spiculated). Mean nodule density and nodule volume were automatically generated by software. Analyses were performed by univariate and multivariate logistic regression. Results were presented as likelihood ratios (LR) with 95% confidence intervals (CI). Receiver operating characteristic analysis was performed for mean density as predictor for lung cancer. RESULTS: Of the 469 nodules, 387 (83%) were between 50 and 500mm3, 82 (17%) >500mm3, 59 (13%) malignant, 410 (87%) benign. The median size of the nodules was 103mm3 (range 50-5486mm3). In multivariate analysis lobulated nodules had LR of 11 compared to smooth; spiculated nodules a LR of 7 compared to smooth; irregular nodules a LR of 6 compared to round and polygonal; volume a LR of 3. The mean nodule CT density did not predict the presence of lung cancer (AUC 0.37, 95% CI 0.32-0.43). CONCLUSION: In solid non-calcified nodules larger than 50mm3, size and to a lesser extent a lobulated or spiculated margin and irregular shape increased the likelihood that a nodule was malignant. Nodule density had no discriminative power. PMID- 17920801 TI - Analysis and stability of the constituents of St. John's wort oils prepared with different methods. AB - St. John's wort is a medicinal plant with a long history of use in traditional medicine all over Europe. Traditional preparations and in particular the infused oil from SJW flowers remains one of the most popular and curative topical remedy against ulcerations and burns. The presence of the characteristic polyprenylated acylphloroglucinol derivatives, namely hyperforin and analogs are instead related to the oil's therapeutic activity. Indeed, it is well known that hyperforin has a potent antibacterial activity. In this study we tried to rationalize the production system of the oily preparation in order to obtain the highest concentration and stability of phloroglucinols. Five different samples of SJW oils were evaluated by HPLC-DAD-MS analysis to verify the variability and stability of the constituents according to the following factors: different harvesting time, fresh or dried plant material, use of sunlight or heating systems during extraction. The stability of these oils during 1 year was also tested. PMID- 17920802 TI - Uptake of ciprofloxacin and ofloxacin by 2 Brucella strains and their fluoroquinolone-resistant variants under different conditions. An in vitro study. AB - The existence of an efflux-mediated mechanism involved in the drug accumulation process of the 2 resistant Brucella strains was investigated; 2 susceptible Brucella strains, Brucella melitensis biovar abortus 2308 (S2308) and AFbortus 3196 (S3196) strains, were the reference strains cultured on trypticase soy agar (TSA) medium and incubated to gradient concentrations of ciprofloxacin and ofloxacin antibiotics. The uptake level of 2 antibacterial agents in 2 susceptible and their fluoroquinolones-resistant variants of Brucella strains obtained in vitro at neutral (pH 7) and acidic (pH 4.5) media, in the presence of carbonyl cyanide m-chlorophenylhydrazone (CCCP), was analyzed spectrophotometrically. A decreased accumulation was found in all strains at the acidic medium. Accumulation of both ciprofloxacin and ofloxacin was found to be lower in resistant strains than in the susceptible ones. Preincubation with the CCCP had an obvious effect in increased intracellular fluoroquinolone fluorescence. The decrease in drug accumulation seen in 2 resistant Brucella strains could be attributed to an energy-depended efflux mechanism that mediated the resistance to fluoroquinolones, which was often associated with porin alterations (resulting to decreased outer membrane permeability). PMID- 17920803 TI - Upstream processes in antibody production: evaluation of critical parameters. AB - The demand for monoclonal antibody for therapeutic and diagnostic applications is rising constantly which puts up a need to bring down the cost of its production. In this context it becomes a prerequisite to improve the efficiency of the existing processes used for monoclonal antibody production. This review describes various upstream processes used for monoclonal antibody production and evaluates critical parameters and efforts which are being made to enhance the efficiency of the process. The upstream technology has tremendously been upgraded from host cells used for manufacturing to bioreactors type and capacity. The host cells used range from microbial, mammalian to plant cells with mammalian cells dominating the scenario. Disposable bioreactors are being promoted for small scale production due to easy adaptation to process validation and flexibility, though they are limited by the scale of production. In this respect Wave bioreactors for suspension culture have been introduced recently. A novel bioreactor for immobilized cells is described which permits an economical and easy alternative to hollow fiber bioreactor at lab scale production. Modification of the cellular machinery to alter their metabolic characteristics has further added to robustness of cells and perks up cell specific productivity. The process parameters including feeding strategies and environmental parameters are being improved and efforts to validate them to get reproducible results are becoming a trend. Online monitoring of the process and product characterization is increasingly gaining importance. In total the advancement of upstream processes have led to the increase in volumetric productivity by 100-fold over last decade and make the monoclonal antibody production more economical and realistic option for therapeutic applications. PMID- 17920804 TI - Luteinizing hormone-dependent Cushing's syndrome in a pet ferret (Mustela putorius furo). AB - Hyperadrenocorticism in ferrets is associated with increased circulating concentrations of adrenal androgens, whereas plasma concentrations of cortisol and ACTH are usually not affected. Here, we report on a 5-year-old castrated male pet ferret (Mustela putorius furo) in which the major presenting signs were polyuria and polyphagia. Routine biochemistry values were within their reference ranges. The urinary corticoid:creatinine ratio (UCCR) was increased and the plasma ACTH concentration was suppressed. Abdominal ultrasonography revealed an enlarged right adrenal gland and atrophy of the left adrenal gland. Administration of hCG resulted in an increase of plasma cortisol and androstenedione concentrations. Based on these findings LH/hCG-dependent hypercortisolism and hyperandrogenism were suspected and treatment was started with a depot GnRH-agonist implant containing 9.4mg deslorelin. Within 3 weeks after placement of the implant all clinical signs had disappeared. Three months later the endocrine parameters had normalized, while abdominal ultrasonography revealed that the right adrenal gland had diminished in size and the left adrenal gland was considered of normal size. No recurrences of clinical signs were seen within 2 years after placement of the deslorelin implant. At that time urinary corticoid and plasma hormone concentrations were within their reference ranges, and no further change in the size of the adrenal glands was seen. In conclusion, this is the first confirmed case of LH-dependent hypercortisolism in a ferret that was treated successfully with a depot GnRH-agonist. PMID- 17920805 TI - [Hepatic encephalopathy in cirrhotic patients admitted in intense care unit]. PMID- 17920806 TI - Titin expression in human articular cartilage and cultured chondrocytes: a novel component in articular cartilage biomechanical sensing? AB - In striated muscle tissues, the giant protein titin acts as a biomechanically active filament system, coupling stress/strain to gene expression. The objective of the study is to show the existence of titin fragments in human articular cartilage, as in diarthodial joints, chondrocytes are also known to sense and respond to stretching. We have surveyed human cultured cartilage collected from adults with osteoarthritis (OA), without OA and from infants with a set of titin antibodies and primer pairs. Three different antibodies were used for immunolabelling, reacting with titin's N-terminal Z1-Z2 domains, its Novex III exon, and with its PEVK region. An antibody directed to a titin ligand was included, since in cardiac muscle, this has been shown to participate in the transmission of stretch dependent titin-based signals. Our results indicate that although at low levels, titin is expressed in cartilage. Primer pairs detected titin transcripts in cartilage, and consistent with this, antibodies directed to titin's Z-disc region and to its elastic region stained cartilage. Moreover, we also could detect transcription of the titin ligand CARP. Components of the stretch dependent signal machinery in muscle are also expressed in cartilage. Further studies are warranted to address if common stress/strain dependent signalling are conserved in muscle and cartilage tissues. PMID- 17920807 TI - The relationship of intelligence to executive function and non-executive function measures in a sample of average, above average, and gifted youth. AB - This study explores the relationship of intelligence to the Wisconsin Card Sorting Test, Stroop Color-Word Test, Oral Word Fluency Test, Design Fluency Test, Trail Making Test, contrasted with Rey Complex Figure Test, Rey Auditory Verbal Learning Test, Wide Range Achievement Test, and Underlining Test in average, above average and gifted children. Full-Scale IQ was significantly related to Wisconsin Card Sort Perseverative and Non-Perseverative Errors, Stroop Color-Word Test, Color-Word condition, Controlled Oral Word Fluency, Design Fluency, Rey Complex Figure, and Underlining conditions but not Trails or Rey Auditory Verbal Learning Test. MANCOVA's show gifted children outperformed other children on the executive but not the non-executive tests. Finally, the nature of the neuropsychological/IQ relationship was explored by further analyses. PMID- 17920808 TI - Behavioral avoidance and self-reported fainting symptoms in blood/injury fearful individuals: an experimental test of disgust domain specificity. AB - This study examined the specificity of disgust in predicting avoidance in blood/injury (BI) phobia. Participants high (n=38) and low (n=46) in BI fear completed measures of disgust across multiple domains and severity of BI-related fear. They then completed three randomly presented behavioral avoidance tasks (BATs) that consisted of exposure to a 15'' severed deer leg (BI task), a live spider (spider task), and a 'contaminated' cookie (cookie task). Fainting symptoms associated with each BAT were recorded as well. When controlling for gender and BI fear group membership, mutilation disgust contributed unique variance to avoidance on the BI task and animal disgust contributed unique variance to avoidance on the spider task. None of the disgust domains contributed unique variance to avoidance on the cookie task. For the high BI fear group, self reported fainting symptoms were more pronounced during the BI and spider BAT than during the cookie BAT. Although mutilation disgust was significantly associated with self-reported fainting symptoms on the BI task among the high BI fear group, this relationship became nonsignificant when controlling for BI-related fear severity. Implications of the domain specificity of disgust and its relevance for understanding fainting responses in BI phobia are discussed. PMID- 17920809 TI - Mutations in the helix termination motif of mouse type I IRS keratin genes impair the assembly of keratin intermediate filament. AB - Two classical mouse hair coat mutations, Rex (Re) and Rex wavy coat (Re(wc)), are linked to the type I inner root sheath (IRS) keratin genes of chromosome 11. An N ethyl-N-nitrosourea-induced mutation, M100573, also maps close to the type I IRS keratin genes. In this study, we demonstrate that Re and M100573 mice bear mutations in the type I IRS gene Krt25; Re(wc) mice bear an additional mutation in the type I IRS gene Krt27. These three mutations are located in the helix termination motif of the 2B alpha-helical rod domain of a type I IRS keratin protein. Immunohistological analysis revealed abnormal foam-like immunoreactivity with an antibody raised to type II IRS keratin K71 in the IRS of Re/+ mice. These results suggest that the helix termination motif is essential for the proper assembly of types I and II IRS keratin protein complexes and the formation of keratin intermediate filaments. PMID- 17920810 TI - Quantitative determination of gene strand bias in prokaryotic genomes. AB - Comparative genometrics of microorganisms is a relatively new area, in which genome properties are translated into numerical indexes. Such indexes can be used for a comprehensive and comparative analysis of microbial genomes, contributing to the understanding of their evolution. This work presents a new method for quantitative determination of gene strand bias in prokaryotic chromosomes, in which data transformation of gene position skew leads to a numerical index that can be applied to quantitative comparisons of genome organization. It was applied in the comparative analysis of 49 completely sequenced Firmicutes genomes, allowing the distinction of groups defined according to their patterns of gene strand preference. The resulting groups revealed that, regarding gene strand bias, reduced genomes are, in general, the more disordered among Firmicutes, while genomes of extremophile organisms comprehend those with the highest degree of genome organization in this phylum. PMID- 17920812 TI - The apical dendrite theory of consciousness. AB - The neural basis of consciousness is theorized here to be the elevated activity of the apical dendrite within a thalamocortical circuit. Both the anatomical and functional properties of these two brain structures are examined within the general context of the cortical minicolumn, which is regarded as the functional unit of the cerebral cortex. Two main circuits of the minicolumn are described: the axis circuit, which sustains activity for extended durations and produces our sensory impressions, and the shell circuit, which performs input-output processing and produces identifications, categorizations, and ideas. The apical dendrite operates within the axis circuit to stabilize neural activity, which enables conscious impressions to be steady and to be sustained over long periods of time. In an attempt to understand how the conscious aspect of subjective impressions may be related to apical dendrite activity, we examine the characteristics of the electric and magnetic fields during the movement of charges along the apical dendrite. The physical correlate of consciousness is regarded here as the relatively intense electromagnetic field that is located along the inside and the outside close to the surface of the active apical dendrite. PMID- 17920811 TI - Glial toll-like receptor signaling in central nervous system infection and autoimmunity. AB - Innate immunity in the CNS depends primarily on the functions of glial cells, astrocytes and microglia, which are important for the early control of pathogen replication and direct the recruitment and activation of cells of the adaptive immune system required for pathogen clearance. Efficient immune responses are required for clearance of an invading pathogen, but dysregulation of a pro inflammatory response in the CNS could lead to the development of autoimmunity. This review summarizes the activation of toll-like receptors (TLRs) expressed on glial cells and the functional outcome of these interactions for CNS health and disease which depends on a delicate balance of the protective and toxic effects of molecules induced in the CNS following TLR ligation. PMID- 17920813 TI - Classification of hyperspectral medical tongue images for tongue diagnosis. AB - Human tongue is one of the important organs of the body, which carries abound of information of the health status. The images of the human tongue that are used in computerized tongue diagnosis of traditional Chinese medicine (TCM) are all RGB color images captured with color CCD cameras currently. However, this conversional method impedes the accurate analysis on the subjects of tongue surface because of the influence of illumination and tongue pose. To address this problem, this paper presents a novel approach to analyze the tongue surface information based on hyperspectral medical tongue images with support vector machines. The experimental results based on chronic Cholecystitis patients and healthy volunteers illustrate its effectiveness. PMID- 17920814 TI - Glenoid and humerus bone analysis using CT transverse sections to automate gleno humeral joint diagnoses and surgery managements. AB - This paper describes an image analysis method that evaluates the glenoid and humerus bone morphology to automate the gleno-humeral (GH) joint diagnoses and surgical managements. This method uses radial B-spline curves to approximate ellipse-like shoulder structures including the humeral stem, tubercle and contact joint as well as the glenoid on every CT transverse section. Radius changes from structure centers to bone boundaries are recognized as convex, concave, separate and hole features that are then identified as pathological spurs, fractures and tumors. The centers and radii of these structures from the transverse sections are integrated to determine the properties of the humeral stem and contact joint with the glenoid, including the stem axis as well as the contact joint and glenoid centers, radii and attitudes. Based on the geometric evaluations of these structures and features, the GH joint surgery including tumor dissect and bone graft, open reduction using screws and plate or nails, and arthroplasty are automatically managed to achieve the normal GH joint functions including dissection of tumors, reduction of fractures or dislocations, and free GH joint motions. This prototype system can be used as a qualitative and quantitative tool for the GH joint diseases diagnoses and surgery managements. A series of examples and case studies illustrate the effectiveness of this automated method. PMID- 17920815 TI - Newspaper reporting on schizophrenia: a content analysis of five national newspapers at two time points. AB - AIM: This study aimed to assess change in the quality of reporting of schizophrenia in UK national daily newspapers, comparing 1996 with 2005. METHODS: Five newspapers were searched using the PROQUEST database for articles published in 1996 or 2005 which contained the term 'schizo...'. 1196 articles were identified and rated against indicators of poor quality reporting. Derived from guidelines for media reporting on mental health the indicators were: metaphoric use of schizophrenia terms; use of stigmatising descriptors; use of equating descriptors; not including information putting the risk of violence into perspective in articles about violence; and use of the term 'release(d)' in articles about discharge from psychiatric hospital. RESULTS: More recent year of publication was associated with a decrease in the odds of an article using a schizophrenia term metaphorically by a factor of 0.103 (95% CI 0.014-0.776), but this masked a pattern of decreasing use in broadsheets and increasing use in tabloids. The use of equating descriptors was significantly lower in 2005 in the univariate analysis (p<0.001), but this was no longer significant (p=0.558) when confounding variables were controlled for. There was no significant change in the use of stigmatising descriptors; the non-inclusion of information putting risk of violence into perspective, nor in the use of the term 'release(d)'. CONCLUSIONS: There is little evidence that the quality of reporting of schizophrenia has changed over time. This suggests a need for the implementation of effective measures to bring newspaper reporting in line with current guidelines. PMID- 17920816 TI - Photoaging and DNA repair. AB - The incidence of sunlight-induced skin changes (photoaged skin, skin carcinogenesis) increases with increasing age and it is thought to be associated with an accumulation of mutations in skin cells. These mutations are mainly caused by UV exposure. The reactive oxygen species produced in UV-exposed skin can cause various kinds of DNA damages e.g., 8-oxoguanine, which are primarily repaired by the base excision repair (BER) system. In addition, UV can directly cause DNA damages; cyclobutane pyrimidine dimers (CPD) and pyrimidine-pyrimidone (6-4) photoproducts (6-4PP), both of which can be repaired by the nucleotide excision repair (NER) system. There have been several reports showing an age related reduction in the DNA repair capacity in the NER, BER, and other repair systems, which contributes to the phenotypes of aging. To clarify the mechanism of skin aging, we examined the NER of skin fibroblasts from healthy donors of different ages. In a host cell reactivation assay, the cells from elderly donors exhibited a significant decline in the ability to restore transfected reporter DNA damaged by UV. In contrast, the ability to remove CPD and 6-4PP declined little with age, as assessed by an enzyme-linked immunosorbent assay. The mRNA expression of DNA repair synthesis-related genes was markedly decreased in the cells from elderly subjects as compared with those from young subjects. These results imply that the age-sensitive step took place after the damage excision in the NER, and that there is an impairment of the latter step of the NER in aging. Based on our data, as well as other reports, the reduced post-UV DNA repair capacity in aging resulting in an accumulation of UV-induced DNA damage is thus considered to be associated with the phenotypes of photoaged skin. PMID- 17920817 TI - Assessment of insertion/deletion polymorphism of the angiotensin converting enzyme gene in psoriasis. PMID- 17920818 TI - Serum levels of IgE anti-BP180 and anti-BP230 autoantibodies in patients with bullous pemphigoid. AB - BACKGROUND: Bullous pemphigoid (BP) is a subepidermal blistering disease characterized by autoantibodies against the hemidesmosomal proteins, BP180 and BP230. NC16A, a non-collagenous stretch of the BP180 ectodomain is the primary target of pathogenic IgG antibodies. Whereas IgG anti-BP180 autoantibodies play a primary role in the pathogenesis, there is a growing number of data regarding the potential pathogenic roles of IgE class autoantibodies in BP. OBJECTIVES: To examine the levels of IgG and IgE autoantibodies against BP180 and BP230, and to investigate mutual association and clinical relevance. METHODS: Sera obtained from 67BP patients and 36 healthy donors were subjected to ELISA assays to measure serum IgG and IgE levels of anti-BP180 and anti-BP230 antibodies. RESULTS: IgG anti-BP180 antibodies were positive in 63 (94%) of 67BP patients. IgG anti-BP230, IgE anti-BP180, and IgE anti-BP230 antibodies were found in 48 (72%), 20 (30%) and 45 (67%), respectively. IgG anti-BP180 levels were correlated with the affected areas. IgG anti-BP230 antibodies tended to increase in proportion to elongation of disease duration. IgE anti-BP230 levels showed a strong association with local eosinophil accumulation, while the levels were reversely related with the affected areas in BP. CONCLUSIONS: IgE autoantibodies to BP180 and BP230 are detected at high frequencies in BP. IgE anti-BP230 antibodies may have a role in attracting eosinophils to the skin lesions. PMID- 17920819 TI - Usefulness of aetiological tests for guiding antibiotic therapy in community acquired pneumonia. AB - The goal with antibiotic therapy in community-acquired pneumonia (CAP) is to cure the patient, ideally without causing side effects and without contributing to the further development of antibiotic resistance. Although patients with severe CAP should be treated with broad-spectrum antibiotics, patients with non-severe CAP should preferably receive pathogen-directed therapy. Rapid aetiological tests, such as sputum Gram stain and urinary antigen tests, are useful for targeting initial pathogen-directed therapy. Non-rapid tests, such as cultures, can subsequently support a switch from initial broad-spectrum therapy to narrow spectrum therapy and direct therapy changes in case of treatment failure. As conventional diagnostic methods often fail to identify the aetiology of CAP, PCR (polymerase chain reaction) tests for respiratory pathogens have become useful and should be further developed. Based on the test specificities, aetiological tests may provide diagnoses with varying reliability, i.e. definite aetiologies (e.g., blood culture and Legionella urinary antigen test), probable aetiologies (e.g., sputum culture and PCR for Mycoplasma pneumoniae), or possible aetiologies (e.g., culture of nasopharyngeal secretions and PCR for Streptococcus pneumoniae). A definite or probable aetiology can often be used to target antibiotic therapy. PMID- 17920820 TI - Trypanocidal activity of piperazine-linked bisbenzamidines and bisbenzamidoxime, an orally active prodrug. AB - A series of 32 piperazine-linked bisbenzamidines (and related analogues) were analysed for their in vitro and in vivo trypanocidal activity against a drug sensitive strain of Trypanosoma brucei brucei and a drug-resistant strain of Trypanosoma brucei rhodesiense. The compounds showed similar potencies against both strains. The most potent compounds were bisbenzamidines substituted at the amidinium nitrogens with a linear pentyl group (8, inhibitory concentration for 50% (IC(50))=1.7-3.0 nM) or cyclic octyl group (17, IC(50)=2.3-4.6 nM). Replacement of the diamidine groups with diamidoxime groups resulted in a prodrug (22) that was effective orally against T. b. brucei-infected mice. Three compounds (7, 11 and 15) provided 100% cure when administered parenterally. The results indicate that the nature of the substituents at the amidinium nitrogens of bisbenzamidines strongly influence their trypanocidal activity. PMID- 17920821 TI - Neuronal correlates of reward and loss in Cluster B personality disorders: a functional magnetic resonance imaging study. AB - Decision making is guided by the likely consequences of behavioural choices. Neuronal correlates of financial reward have been described in a number of functional imaging studies in humans. Areas implicated in reward include ventral striatum, dopaminergic midbrain, amygdala and orbitofrontal cortex. Response to loss has not been as extensively studied but may involve prefrontal and medial temporal cortices. It has been proposed that increased sensitivity to reward and reduced sensitivity to punishment underlie some of the psychopathology in impulsive personality disordered individuals. However, few imaging studies using reinforcement tasks have been conducted in this group. In this fMRI study, we investigate the effects of positive (monetary reward) and negative (monetary loss) outcomes on BOLD responses in two target selection tasks. The experimental group comprised eight people with Cluster B (antisocial and borderline) personality disorder, whilst the control group contained fourteen healthy participants. A key finding was the absence of prefrontal responses and reduced BOLD signal in the subcortical reward system in the PD group during positive reinforcement. Impulsivity scores correlated negatively with prefrontal responses in the PD but not the control group during both, reward and loss. Our results suggest dysfunctional responses to rewarding and aversive stimuli in Cluster B personality disordered individuals but do not support the notion of hypersensitivity to reward and hyposensitivity to loss. PMID- 17920822 TI - Enhanced bioavailability of metoclopramide HCl by intranasal administration of a mucoadhesive in situ gel with modulated rheological and mucociliary transport properties. AB - The prolonged residence of drug formulation in the nasal cavity is of utmost importance for intranasal drug delivery. The objective of the present investigation was to develop a mucoadhesive in situ gel with reduced nasal mucociliary clearance in order to improve the bioavailability of the antiemetic drug, metoclopramide hydrochloride (MCP HCl). The in situ gelation upon contact with nasal mucosa was conferred via the use of the thermogelling poloxamer 407 whereas mucoadhesion and drug release enhancement were modulated via the use of mucoadhesive and polyethylene glycol (PEG) polymers respectively. The results revealed that the different mucoadhesives augmented the gel viscosity but reduced its sol-gel transition temperatures (T(sol-gel)) and the drug release. The inclusion of PEG counteracted the effect of the mucoadhesive polymers whereby it decreased the gel consistency and increased the T(sol-gel) as well as the in vitro drug release. The formulations with favorable sol-gel transition temperatures (25-32 degrees C) and high in vitro drug release (100% release in 60 min) were also rheologically stable upon storage. The mucoadhesiveness test was performed in vivo in rats, results showed that the carbopol-containing in situ gel prolonged the mucociliary transport time from 10 min (control solution) to 52 min (mucoadhesive gel) and maintained nasal mucosal integrity after 14-days application. The bioavailability study in rabbits revealed that the absolute bioavailability of MCP HCl was significantly increased from 51.7% in case of the oral drug solution to 69.1% in case of the nasal in situ gel. The study point to the potential of mucoadhesive nasal in situ gel in terms of ease of administration, accuracy of dosing, prolonged nasal residence and improved drug bioavailability. PMID- 17920823 TI - [A multidisciplinary consultation for children with brain tumors]. AB - School achievement of children with brain tumors is hampered by progressive neurologic and cognitive sequelae. To help the children and their family, we have created in 1997 a multidisciplinary consultation together with Necker's hospital. MATERIAL AND METHODS: The study describes the organization of the consultation and analyses the files of 69 children seen between September 2001 and June 2002. RESULTS AND CONCLUSION: The authors conclude that this consultation is an irreplaceable mean to coordinate the complex rehabilitation process of a child treated for a brain tumor. PMID- 17920824 TI - [MRI finding in myxopapillary ependymoma of the filum terminal: a case report]. PMID- 17920825 TI - Intervention by phone calls raises domiciliary activity and exercise capacity in patients with severe COPD. AB - BACKGROUND: Patients with severe COPD suffer from impairments of exercise capacity which affects daily activity. Conversely, activity might exert effects on the functional state. We studied whether a short-term intervention by regular phone calls caused an increase in activity at home and whether this resulted in a gain in exercise capacity. METHODS: Over a 2-week period (P1) normal daily activity was assessed in 21 patients with stable severe COPD (GOLD III/IV). After this, the individual setting was explored in a short home visit. The subsequent 2 week period (P2) involved phone calls every other day to raise home-based activity (target: 3x15 min daily at 75% of maximum dyspnea). During the study, patients wore an actograph plus pedometer and kept a diary. Before P1 and after P2, 6-min walking distance (6MWD), lung function, the Borg score and quality of life (SF-36, SGRQ) were determined. RESULTS: Compared to P1, actograph counts (p<0.05) were higher in P2. There was also an increase in 6MWD (p<0.05) and quality of life scores (SF-36, p<0.05) between initial and final visit, whereby improvements in 6MWD correlated with changes in activity (p<0.01). Conversely, four patients who experienced an exacerbation in P2 showed no increase in activity or 6MWD. CONCLUSIONS: In patients with stable severe COPD, it was possible to increase activity by regular phone calls without performing previous rehabilitation. Increased activity resulted in increased exercise capacity and quality of life within 2 weeks, underlining the effectiveness of continued motivational support in patients with severe COPD. PMID- 17920827 TI - Experimental evaluation of fog warning system. AB - Highway safety is a major concern to the public and to transportation professionals, so the number of crashes caused by poor visibility due to fog form an alarming statistic. Drivers respond to poor visibility conditions in different ways: some slow down; others do not. Many drivers simply follow the taillights of the vehicle ahead. Accordingly, hazardous conditions are created in which speeds are both too high for the prevailing conditions and highly variable. Findings are presented from a study of traffic crashes due to fog in the southern region of Saudi Arabia. The primary objective was to assess the effectiveness of fog detection and warning system on driver behavior regarding speed and headway. This warning system includes visibility sensors that automatically activate a variable message sign that posts an advisory speed when hazardous conditions due to fog occur. The system was installed on a 2 km section of a two-lane, rural highway. A data set of 36,013 observations from both experimental and control sections at two study sites was collected and analyzed. The data included vehicle speed, volume, and classification; time headway, time of day, and visibility distance. Although the warning system was ineffective in reducing speed variability, mean speed throughout the experimental sections was reduced by about 6.5 kph. This reduction indicates that the warning system appeared to have a positive effect on driver behavior in fog even though the observed mean speeds were still higher than the posted advisory speed. From relationships found in the literature between mean driving speed and number of crashes, a speed reduction of only 5 kph would yield a 15% decrease in the number of crashes. PMID- 17920826 TI - The epidemiology of GHB and ketamine use in an Australian household survey. AB - BACKGROUND: There have been apparent increases in recent years in the illicit use of ketamine and gamma-hydroxybutyrate (GHB), but to date there has been no examination of the epidemiology of use in the general population. This paper provides the first such Australian data on the patterns and correlates of GHB and ketamine use. METHOD: Data were analysed from the 2004 National Drug Strategy Household Survey, a multistage probability sample of Australians aged 14 years or older. Associations between GHB and ketamine use, and core demographic and other drug use variables, were examined. RESULTS: 0.5% of Australians aged 14 years or older reported ever using GHB, and 0.1% reported recent use, with the prevalence of use being highest amongst those aged 20-29 years. Lifetime use of ketamine was reported by 1% of Australians aged 14 years or older, with 0.3% reporting recent use. Again, prevalence of ketamine use was highest amongst those aged 20-29 years. Those who reported ever using these drugs described a pattern of occasional use, with the large majority not using these drugs in the past year. Multiple regression analyses suggested that compared to non-users, GHB and ketamine users were more likely to report the recent use of a wide range of other drugs. CONCLUSIONS: The prevalence of GHB and ketamine use in Australia appears to be quite low. The present study found high rates of polydrug use, as have been documented in convenience samples of GHB and ketamine users in previous work. As for other illegal drugs used by small proportions of the population, detailed data on patterns of use and associated risks of use are probably best derived from targeted samples of users; household survey data allow comparisons of the relative prevalence of use compared to other illicit drugs and future work will provide the opportunity to consider changes in the extent of use in the general population. PMID- 17920828 TI - Crash risk of older drivers after attending a mature driver education program. AB - PURPOSE: The primary aim of this study was to determine if the crash rate of aging drivers can be mitigated by post-license driver education. DESIGN AND METHODS: This study of 884 older drivers who attended the 55 Alive/Mature Driving program was conducted in three phases. Phase 1, which examined self-selection bias of seniors attending the driver education program, and Phase 2, which examined changes in crash rate after attending the program, were carried out through analysis of driving records before and after attending the course. In Phase 3, the use of selection, optimization, and compensation strategies by older male drivers who attended 55 Alive/Mature Driving was addressed through focus group interviews. RESULTS: Findings showed a self-selection bias among older drivers who attended 55 Alive/Mature Driving. Results also showed attendance at the program was associated with an increased number of crashes for men aged 75 years and older, but no effect on subsequent crashes of younger men and women of all ages. Focus group sessions suggested older men who attended the program used fewer strategies to cope with their declining skills. IMPLICATIONS: Recognizing and understanding characteristics and behaviors of older drivers who attend remedial driver education is essential to the design and delivery of successful driver safety programs. PMID- 17920829 TI - Prevalence of impaired driving behaviors in a diverse, rural, southern middle school. AB - Little research has been reported on the risky behaviors of pre-driving adolescents revolving around alcohol use, in particular impaired driving behaviors, which in general have been shown to be higher in rural areas. This study investigated the prevalence of drinking while driving and riding with a drinking driver among 290 middle school students in a Mississippi Delta area middle school. Just under half (45.3%) responded that at least once in the past 30 days they had ridden with a drinking driver and 17% indicated they had driven an automobile after drinking alcohol. The prevalence of underage drinking, driving under the influence of alcohol, and riding with a drinking driver among our sample of middle school students is alarming. This study shows alcohol related driving behaviors are not solely performed by those who are legally licensed to drive, but simply by those who have access to vehicles. The authors recommend that prevention programs focusing on reducing the incidence of impaired driving should start in early adolescence. PMID- 17920830 TI - Roles of safety climate and shift work on perceived injury risk: a multi-level analysis. AB - This study evaluated the relationship between employees' work shift (i.e., day shift versus night shift) and perceptions of injury risk, and how the relationship is affected by company level safety climate and injury frequency. The results showed that night shift workers perceived a higher level of injury risk compared to day shift workers. Both company level safety climate and injury frequency played critical roles in predicting individual perceived work injury risk. Perception of injury risk of night shift workers was significantly lower when they perceived high-level rather than low-level safety climate. However, this pattern was not noticeable for day shift workers. These findings highlighted the importance of considering company level factors when attempting to understand the differences between day shift and night shift work on an individual's perception of injury risk. PMID- 17920831 TI - Attitudes of commercial motor vehicle drivers towards safety belts. AB - Despite the fact that Hawaii has one of the highest seat belt use rates for passenger vehicles in the United States, and has had a mandatory seat belt use law since the 1980s, studies have shown that commercial motor vehicles (CMV) seat belt use rates are low. To better understand this phenomenon, a comprehensive survey of commercial vehicle drivers was conducted in Hawaii to ascertain attitudes and self-reported behaviors regarding seat belt use. A total of 791 drivers responded to a written questionnaire implemented at weigh stations and distributed to various trucking firms and transport centers. Approximately 67% reported that they use seat belts "always" when driving a CMV (commercial motor vehicle), yet when asked how often do other CMV drivers use seat belts, only 31% responded "always." Interestingly, 86% of these same drivers reported that they use seat belts "always" when driving a personal vehicle. The major reason cited for non-use of belts was "frequent stops/inconvenience" (29%), and "not safety conscious" (23%). Notably, the self-reported use of safety belts is highest among operators of vans (88% said "always"), followed by buses (87% said "always") and lowest among truck drivers (only 60% said "always"). In this paper, some of the differences between self-reported users and non-users are explored and a multivariate logit model was developed to predict the odds of belt use as a function of various factors. PMID- 17920832 TI - Contributory factors to traffic crashes at signalized intersections in Hong Kong. AB - Efficient geometric design and signal timing not only improve operational performance at signalized intersections by expanding capacity and reducing traffic delays, but also result in an appreciable reduction in traffic conflicts, and thus better road safety. Information on the incidence of crashes, traffic flow, geometric design, road environment, and traffic control at 262 signalized intersections in Hong Kong during 2002 and 2003 are incorporated into a crash prediction model. Poisson regression and negative binomial regression are used to quantify the influence of possible contributory factors on the incidence of killed and severe injury (KSI) crashes and slight injury crashes, respectively, while possible interventions by traffic flow are controlled. The results for the incidence of slight injury crashes reveal that the road environment, degree of curvature, and presence of tram stops are significant factors, and that traffic volume has a diminishing effect on the crash risk. The presence of tram stops, number of pedestrian streams, road environment, proportion of commercial vehicles, average lane width, and degree of curvature increase the risk of KSI crashes, but the effect of traffic volume is negligible. PMID- 17920833 TI - Mild neuritic changes are increased in the brains of fatally injured older motor vehicle drivers. AB - Given the expected increase in the older population and driving in this age group, concerns have been raised about the safety of older drivers. People over 65 years are over-represented in motor vehicle fatalities when calculated by distance driven. They are also at risk of neurodegenerative diseases, such as Alzheimer's disease, that affect cognitive function. We have examined the brains of older drivers (15M:12F) who died as a result of a motor vehicle accident (MVA) to determine the extent of Alzheimer's disease-related neurofibrillary changes (neuritic plaques and neurofibrillary tangles), Lewy body pathology and cerebrovascular disease and compared them to a control group of older licenced drivers (23M:5F) who died of other causes. The prevalence of moderate or severe neuritic plaque pathology was less than expected for the general population of this age and there was no difference between the groups. However, mild neuritic plaque pathology was increased for MVA deaths compared to controls. There was no evidence of vascular dementia or dementia with Lewy bodies. The current mandatory age-related re-licencing procedures in NSW may contribute to the low percentage of drivers with severe pathology. Further research into the role of mild pathology in cognitive impairment and older drivers is warranted. PMID- 17920834 TI - The credibility of speed limits on 80 km/h rural roads: The effects of road and person(ality) characteristics. AB - The present study aimed to operationalise the concept of credible speed limits and to make a first step to elaborate it in such a way that road authorities can put the concept into practice. The study focused on the credibility of an 80 km/h limit for different rural roads and assessed the effects of characteristics of the road and its environment as well as the effects of person and personality characteristics. Almost 600 Dutch car drivers were asked to judge 27 photographs of (different) rural roads with a posted speed limit of 80 km/h. To determine the degree of credibility, for each road scene the subjects filled in the preferred speed and the speed limit they considered to be safe. The results show large differences in both preferred speed and the safe speed limit between the road scenes, both below and above the limit of 80 km/h. These differences were related to a number of characteristics of the road and the road environment, such as the presence or absence of a curve and characteristics concerning the field of view (sight distance, clarity of situation). There were also large differences between subjects and these were related to age, the degree of sensation seeking, the number of speeding tickets in the last 3 years and the part of the country they lived in. Subjects were influenced by more or less the same road features. PMID- 17920835 TI - Do states upgrading to primary enforcement of safety belt laws experience increased daytime and nighttime belt use? AB - As of 1 January 2007, 26 states and the District of Columbia have enacted primary enforcement of their safety belt laws, which allows law enforcement to stop motorists and cite them solely when they observe a vehicle occupant who is not wearing a safety belt. Interrupted time series analyses were used to determine whether six states which upgraded to primary enforcement laws experienced changes in nighttime (9:00 p.m. to 4:59 a.m.) and daytime (5:00 a.m. to 8:59 p.m.) safety belt use based on proxy estimates from fatal crash-involved vehicle occupants. Nighttime and daytime safety belt use increased in five of the six states after the primary enforcement laws were enacted. Because the methods used in these analyses reduced the likelihood that these increases resulted from preexisting secular trends towards increased belt use, the results provide strong support that upgrading from secondary to primary enforcement increases occupant safety belt use during both daytime and nighttime periods. PMID- 17920836 TI - The sleep of commercial vehicle drivers under the 2003 hours-of-service regulations. AB - Previous research has found that commercial drivers get an average of 5.18 h of sleep per night. The revised hours-of-service (HOS) regulations (in the United States) are in place to provide drivers with more opportunities to get sleep. However, are drivers really getting more sleep under these new regulations? Also, is there a relationship between sleep quantity and involvement in critical incident (crashes, near-crashes, or crash-relevant conflicts)? Data from 73 truck drivers, collected during a naturalistic driving study after the implementation of the 2003 HOS regulations, were analyzed to determine overall sleep quantity (using actigraphy), along with sleep quantity prior to being involved in a critical incident. Sixty-two drivers had at least seven consecutive days (Monday through Sunday) of reliable actigraphy data; mean sleep quantity per 24-h period (midnight centered using the Cole-Kripke algorithm) for these drivers was 6.28 h (S.D.=1.42 h). Fifty-eight critical incidents were recorded in the 10th and 11th driving hours. Analysis results indicated that drivers received significantly less sleep in the period prior to a critical incident as compared to their mean overall sleep quantity. The results of this study indicate drivers may be getting more sleep under the revised 2003 HOS regulations as compared to the old regulations. In addition, significantly less sleep in the 24-h period prior to involvement in a critical incident suggests driver fatigue may have been a potential contributing factor in these critical incidents. PMID- 17920837 TI - Impact of BAC limit reduction on different population segments: a Poisson fixed effect analysis. AB - Over the past few decades, several countries enacted the reduction of the legal blood alcohol concentration (BAC) limit, often alongside the administrative license revocation or suspension, to battle drinking-and-driving behavior. Several researchers investigated the effectiveness of these policies by applying different analysis procedures, while assuming population homogeneity in responding to these laws. The present analysis focuses on the evaluation of the impact of BAC limit reduction on different population segments. Poisson regression models, adapted to account for possible observation dependence over time and state specific effects, are estimated to measure the reduction of the number of alcohol-related accidents and fatalities for single-vehicle accidents in 22 U.S. jurisdictions over a period of 15 years starting in 1990. Model estimates demonstrate that, for alcohol-related single-vehicle crashes, (i) BAC laws are more effective in terms of reduction of number of casualties rather than number of accidents, (ii) women and elderly population exhibit higher law compliance with respect to men and to young adult and adult population, respectively, and (iii) the presence of passengers in the vehicle enhances the sense of responsibility of the driver. PMID- 17920838 TI - Explicit and implicit self-enhancement biases in drivers and their relationship to driving violations and crash-risk optimism. AB - Previous research has found that drivers tend to consider themselves superior to their peers on both driving ability and driving caution, as well as judging themselves as at less risk of a crash (crash-risk optimism). These studies have relied on explicit measures by getting drivers to respond to written items. The current study measured 158 New Zealand drivers' explicit and implicit attitudes towards their own driving attributes in comparison with others. Implicit attitudes were measured using a computer-based reaction time task, the Implicit Association Test (IAT). Both explicit and implicit self-enhancement biases were found in driver ability and driver caution. Implicit biases were considerably stronger than explicit biases and men demonstrated stronger self-enhancement biases in driving ability than women. Explicit and implicit ratings of driving ability and explicit ratings of driver caution predicted crash-risk optimism. Explicit and implicit ratings of driving caution predicted a measure of driving violations. The implications for safety interventions and research on drivers' mental processes are discussed particularly in regard to the ability of implicit measures to bypass social desirability effects. PMID- 17920839 TI - Total costs of bicycle injuries in Norway: correcting injury figures and indicating data needs. AB - Bicycle injuries and fatalities are reported by the police to Statistics Norway. Fatality records from the police are then corrected with Vital Statistics records. However, there is no complete hospital recording that could provide more correct data for bicycle injuries. Bicycle injuries are underreported in official data. There is a nearly complete omission of single bicycle accidents. This disguises societal accident costs and curtails the identification of black spots and effective infrastructure improvements. This paper provides an estimate of total bicycle injuries in Norway and the total costs of these injuries. Application of case study hospital data from Norwegian towns enabled an estimation of the relationship between these data and the official data, including the distribution of injuries by severity. Costs were then assessed by applying official monetary values for given levels of injury severity. Total annual bicycle injury costs are huge, but these costs must be balanced against the benefits of bicycling, related to health and environment. Accident reporting and data should be enhanced to enable a reduction of bicycle injuries. PMID- 17920840 TI - The contribution of passengers versus mobile phone use to motor vehicle crashes resulting in hospital attendance by the driver. AB - There is evidence that mobile phone use while driving (including hands-free) is associated with motor vehicle crashes. However, whether the effects of mobile phone use differ from that of passengers in the vehicle remains unclear. The aim of this research was to estimate the risk of crash associated with passenger carriage and compare that with mobile phone use. A case-control study ('passenger study') was performed in Perth, Western Australia in 2003 and 2004. Cases were 274 drivers who attended hospital following a motor vehicle crash and controls were 1096 drivers (1:4 matching) recruited at service stations matched to the location and time and day of week of the crash. The results were compared with those of a case-crossover study ('mobile phone study') undertaken concurrently (n=456); 152 cases were common to both studies. Passenger carriage increased the likelihood of a crash (adjusted odds ratio (adj. OR), 95% confidence interval (95% CI), 1.6, 1.1-2.2). Drivers carrying two or more passengers were twice as likely to crash as unaccompanied drivers (adj. OR 2.2, 95% CI 1.3-3.8). By comparison, driver's use of a mobile phone within 5 min before a crash was associated with a fourfold increased likelihood of crashing (OR 4.1, 95% CI 2.2 7.7). Passenger carriage and increasing numbers of passengers are associated with an increased likelihood of crash, though not to the same extent as mobile phone use. Further research is needed to investigate the factors underlying the increased risks. PMID- 17920841 TI - Using fuzzy signal detection theory to determine why experienced and trained drivers respond faster than novices in a hazard perception test. AB - Drivers' hazard perception ability, as measured in video-based simulations, correlates with crash involvement, improves with experience and can be trained. We propose two alternative signal detection models that could describe individual differences in this skill. The first model states that novice drivers are poorer at discriminating more hazardous from less hazardous situations than experienced drivers. The second model proposes that novice drivers require a higher threshold of danger to be present before they notice a situation is hazardous or before they are willing to classify a situation as hazardous. We applied a technique involving fuzzy signal detection analysis to differentiate between these two models when comparing novice and experienced drivers, and trained and untrained drivers, in various video-based hazard perception measures. The data favored the second model. PMID- 17920842 TI - Storage and utilization patterns of cleaning products in the home: toxicity implications. AB - BACKGROUND AND AIMS: Cleaning products are used to achieve cleaner homes. However, they have been responsible for a considerable number of poisoning cases. The aims of this study were to investigate the storage and utilization patterns of cleaning products in the home, as well as the risk of adverse and toxic effects produced by them. METHODS: This study has been performed using a questionnaire that was developed by the Poison Control and Drug Information Center (PCDIC). The questionnaire was distributed randomly in northern Palestine. The questionnaire included questions regarding storage, utilization habits, and the adverse effects experienced by respondents upon handling the cleaning products. RESULTS: All respondents utilized and stored cleaning products in their homes. Chlorine bleach and acidic cleaning products were the most common. Respondents stored cleaning products at different places in their homes, but most of those storage places were suboptimal and were within the reach of children. The daily utilization rate of cleaning products was 1.6+/-0.8 with chlorine bleach being the most commonly used. Of the respondents, 27% reported experiencing a wide range of acute adverse and toxic effects resulting from cleaning product use and that exposure occurred mainly via inhalation. Caustic substances, bleach, and kerosene were the agents mainly involved in producing these adverse and toxic effects. About half of those who experienced adverse and toxic effects sought medical help, and 22% of them were children younger than 6 years of age. Finally, most respondents reported mixing and discarding cleaning product leftovers and their containers improperly. CONCLUSION AND RECOMMENDATIONS: Correct utilization and safer storage of cleaning products is encouraged. Several preventive strategies should be implemented in order to decrease the incidence of accidental harmful exposure that is due to cleaning agents. The role of the PCDIC is very important in the education, prevention, and management of cleaning product-induced adverse effects. PMID- 17920843 TI - Bayesian multiple testing procedures for hotspot identification. AB - Ranking a group of candidate sites and selecting from it the high-risk locations or hotspots for detailed engineering study and countermeasure evaluation is the first step in a transport safety improvement program. Past studies have however mainly focused on the task of applying appropriate methods for ranking locations, with few focusing on the issue of how to define selection methods or threshold rules for hotspot identification. The primary goal of this paper is to introduce a multiple testing-based approach to the problem of selecting hotspots. Following the recent developments in the literature, two testing procedures are studied under a Bayesian framework: Bayesian test with weights (BTW) and a Bayesian test controlling for the posterior false discovery rate (FDR) or false negative rate (FNR). The hypotheses tests are implemented on the basis of two random effect or Bayesian models, namely, the hierarchical Poisson/Gamma or Negative Binomial model and the hierarchical Poisson/Lognormal model. A dataset of highway-railway grade crossings is used as an application example to illustrate the proposed procedures incorporating both the posterior distribution of accident frequency and the posterior distribution of ranks. Results on the effects of various decision parameters used in hotspot identification procedures are discussed. PMID- 17920844 TI - An evaluation of a new instrument to measure organisational safety culture values and practices. AB - The main aim of this research is to evaluate a safety culture measuring instrument centred upon relevant organisational values and practices related to the safety management system. Seven dimensions that reflect underlying safety meanings are proposed. A second objective is to explore the four cultural orientations in the field of safety arising from the competing values framework. The study sample consisted of 299 participants from five companies in different sectors. The results show six dimensions of organisational values and practices and different company profiles in the organisations studied. The four cultural orientations proposed by the competing values framework are not confirmed. Nevertheless, a coexistence of diverse cultural orientations or paradoxes in the companies is observed. PMID- 17920845 TI - The impact of red light cameras on safety in Arizona. AB - Red light cameras (RLCs) have been used in a number of US cities to yield a demonstrable reduction in red light violations; however, evaluating their impact on safety (crashes) has been relatively more difficult. Accurately estimating the safety impacts of RLCs is challenging for several reasons. First, many safety related factors are uncontrolled and/or confounded during the periods of observation. Second, "spillover" effects caused by drivers reacting to non-RLC equipped intersections and approaches can make the selection of comparison sites difficult. Third, sites selected for RLC installation may not be selected randomly, and as a result may suffer from the regression to the mean bias. Finally, crash severity and resulting costs need to be considered in order to fully understand the safety impacts of RLCs. Recognizing these challenges, a study was conducted to estimate the safety impacts of RLCs on traffic crashes at signalized intersections in the cities of Phoenix and Scottsdale, Arizona. Twenty four RLC equipped intersections in both cities are examined in detail and conclusions are drawn. Four different evaluation methodologies were employed to cope with the technical challenges described in this paper and to assess the sensitivity of results based on analytical assumptions. The evaluation results indicated that both Phoenix and Scottsdale are operating cost-effective installations of RLCs: however, the variability in RLC effectiveness within jurisdictions is larger in Phoenix. Consistent with findings in other regions, angle and left-turn crashes are reduced in general, while rear-end crashes tend to increase as a result of RLCs. PMID- 17920846 TI - Road environment, crash type and hospitalisation of bicyclists and motorcyclists presented to emergency departments in Western Australia. AB - Emergency presentations by bicyclists and motorcyclists are often the result from a fall or non-motor vehicle collision that occurred in off-road locations. Consequently, they are unlikely to be captured by police records. If the injury is not severe enough to warrant hospitalisation, they will also not be captured by the hospital admission system. To ascertain the nature and type of these crash events, a 6-month prospective study was undertaken of bicyclists and motorcyclists who presented to emergency departments in Perth, Western Australia, due to involvement in a crash or non-motor vehicle collision. Of the 330 eligible presentations, 151 bicyclists and 104 motorcyclists agreed to participate in a structured interview, representing a response rate of 77.3%. Among them, 120 (79%) bicyclists and 71 (68%) motorcyclists had a fall or non-motor vehicle collision, and many of the crashes (88 (58%) and 47 (45%), respectively) occurred off-road. Moreover, 26.5% of bicyclist and 62.5% of motorcyclist presentations led to hospital admissions. Multivariate logistic regression analysis further showed that the crash location and road type affected hospitalisation for both groups. Although, crashes occurring in rural areas contributed 14% of the events overall, their adjusted risk of hospitalisation increased five times when compared to metropolitan locations. Crash preventive measures targeting rural areas should be considered to further improve the safety of bicyclists and motorcyclists. PMID- 17920847 TI - Measuring accident risk exposure for pedestrians in different micro-environments. AB - Pedestrians are mainly exposed to the risk of road accident when crossing a road in urban areas. Traditionally in the road safety field, the risk of accident for pedestrian is estimated as a rate of accident involvement per unit of time spent on the road network. The objective of this research is to develop an approach of accident risk based on the concept of risk exposure used in environmental epidemiology, such as in the case of exposure to pollutants. This type of indicator would be useful for comparing the effects of urban transportation policy scenarios on pedestrian safety. The first step is to create an indicator of pedestrians' exposure, which is based on motorised vehicles' "concentration" by lane and also takes account of traffic speed and time spent to cross. This is applied to two specific micro-environments: junctions and mid-block locations. A model of pedestrians' crossing behaviour along a trip is then developed, based on a hierarchical choice between junctions and mid-block locations and taking account of origin and destination, traffic characteristics and pedestrian facilities. Finally, a complete framework is produced for modelling pedestrians' exposure in the light of their crossing behaviour. The feasibility of this approach is demonstrated on an artificial network and a first set of results is obtained from the validation of the models in observational studies. PMID- 17920848 TI - Seasonal patterns of fatal and nonfatal falls among older adults in the U.S. AB - INTRODUCTION: While many believe that older adults fall more often during the winter months, research on this is inconclusive. This study used nationally representative data from 2001 to 2002 to examine unintentional fatal fall rates among older men and women by season and climate, and nonfatal fall rates by season. METHODS: We studied fatal and nonfatal unintentional falls among U.S. adults aged > or =65 during December 2001-November 2002 by season. Fatal fall data were obtained from National Center for Health Statistics' annual mortality tapes; nonfatal fall data for injuries treated in emergency departments were obtained from the National Electronic Injury Surveillance System All Injury Program. Fatal falls were also analyzed by climate based on each state's average January 1, 2001 temperature (colder climates < or =32 degrees F (0 degrees C) and warmer climates >32 degrees F (0 degrees C)). RESULTS: From December 2001 through November 2002, neither fatal nor nonfatal fall injury rates showed any seasonal patterns. For fatal falls, the average rate was 9.1 percent higher in colder climates, regardless of season. CONCLUSION: Among older adults, fatal fall rates appear to be influenced more by climate than by season. Additional research is needed to clarify the mechanisms underlying these observations. PMID- 17920849 TI - The guessability of traffic signs: effects of prospective-user factors and sign design features. AB - This experiment investigated the relationships between the characteristics of prospective-users of traffic signs (people who will use the signs in the future) and the guessability of traffic signs, and also examined the effects of sign design features on the guessability of traffic signs. Forty-one Hong Kong Chinese subjects guessed the meanings and rated the sign features of 120 Mainland Chinese signs. Contrary to expectation, cycling experience and previous experience with sign information had no effect on sign guessing. Males and females with similar education level had similar guessing performance. Previous experience of visiting Mainland China was a significant predictor of guessing performance. Family ownership of a vehicle was associated with guessing performance for subjects who intended to become a driver and for those with car game experience. Subjects who claimed to pay attention to traffic signs in daily life performed better at sign guessing than those who did not. Traffic incident experience did not seem to enhance awareness of, or knowledge about, traffic signs. Guessability of a sign varied with the five design features of; familiarity, concreteness, simplicity, meaningfulness, and semantic closeness of the sign. Semantic closeness was the best predictor of guessability score, followed by familiarity, meaningfulness, concreteness, and simplicity. In order to design more user-friendly traffic signs and effective ways of using them, it is suggested that designers develop and evaluate signs according to the relative importance of the five sign features. PMID- 17920850 TI - Costs of occupational injuries in construction in the United States. AB - This paper presents costs of fatal and nonfatal injuries for the construction industry using 2002 national incidence data from the Bureau of Labor Statistics and a comprehensive cost model that includes direct medical costs, indirect losses in wage and household productivity, as well as an estimate of the quality of life costs due to injury. Costs are presented at the three-digit industry level, by worker characteristics, and by detailed source and event of injury. The total costs of fatal and nonfatal injuries in the construction industry were estimated at $11.5 billion in 2002, 15% of the costs for all private industry. The average cost per case of fatal or nonfatal injury is $27,000 in construction, almost double the per-case cost of $15,000 for all industry in 2002. Five industries accounted for over half the industry's total fatal and nonfatal injury costs. They were miscellaneous special trade contractors (SIC 179), followed by plumbing, heating and air-conditioning (SIC 171), electrical work (SIC 173), heavy construction except highway (SIC 162), and residential building construction (SIC 152), each with over $1 billion in costs. PMID- 17920851 TI - Diagnostic analysis of the logistic model for pedestrian injury severity in traffic crashes. AB - This study attempts to evaluate the injury risk of pedestrian casualties in traffic crashes and to explore the factors that contribute to mortality and severe injury, using the comprehensive historical crash record that is maintained by the Hong Kong Transport Department. The injury, demographic, crash, environmental, geometric, and traffic characteristics of 73,746 pedestrian casualties that were involved in traffic crashes from 1991 to 2004 are considered. Binary logistic regression is used to determine the associations between the probability of fatality and severe injury and all contributory factors. A consideration of the influence of implicit attributes on the trend of pedestrian injury risk, temporal confounding, and interaction effects is progressively incorporated into the predictive model. To verify the goodness-of fit of the proposed model, the Hosmer-Lemeshow test and logistic regression diagnostics are conducted. It is revealed that there is a decreasing trend in pedestrian injury risk, controlling for the influences of demographic, road environment, and other risk factors. In addition, the influences of pedestrian behavior, traffic congestion, and junction type on pedestrian injury risk are subject to temporal variation. PMID- 17920852 TI - Home and leisure injuries among the French electricity and gas company active employees: circumstances and short-term consequences. AB - OBJECTIVE: To characterize home and leisure injuries and their immediate consequences among adults aged 20-60 years and to look for homogeneous profiles of injury circumstances to assess the possibility of setting up prevention programs. DESIGN: Cross-sectional survey by questionnaire completed on the occasion of a medical visit after a first episode of absence for home or leisure injury between 1 January and 31 December 1997 among Electricite de France Gaz de France (EDF GDF) workforce of three geographical areas (47,681 employees). Incidence and relative risks according to sex, age and work grade and a multidimensional classification of injury circumstances. RESULTS: Eight hundred and fifty four injuries were studied. Risk was estimated at 18.4 injury victims per 1000 employees, of which 13.4 per 1000 employees were home injury victims and 3.7 leisure injury victims. The risk of all injuries was higher among men than women (RR=1.3) and decreased as work grade rose: for men, it was five times higher among operating employees than managers. Multidimensional analysis of injury circumstances ended by distributing into four main classes which may be useful for prevention: gardening and do-it-yourself injuries outdoors (19.9%), coming and going on the streets (6.9%), falling while coming and going in the home on the stairs (13.4%), and do-it-yourself inside the home (13.0%). Injuries induced essentially four types of lesions: sprains (34%), fractures (31.8%), contusions (24.5%) and wounds. The mean sick-leave lasted 32.1 days but half the subjects returned to work in less than 17 days. Hospitalization was necessary in 19.2% of cases. CONCLUSIONS: This study completed by an analysis of the behavioural factors of injuries led us to propose programs aimed at changing the risk behaviours related to do-it-yourself, stairs falls and gardening. PMID- 17920853 TI - Effort-reward imbalance at work and driving anger in an Australian community sample: is there a link between work stress and road rage? AB - Both workplace stress and road rage are reported to be on the increase. This study examined the effort-reward imbalance (ERI) model of work stress and its relationship with general anger and driving anger in a community sample of 130 Australian workers. It also examined international differences in driving anger, with Australian motorists reporting lower levels of driving anger than American motorists but higher levels than British motorists. Hierarchical multiple regressions confirmed ERI increased driving anger via the mediating variables of general anger and overcommitment; individuals suffering ERI may develop increased general anger or overcommitment, in turn increasing propensity to experience driving anger. Regressions also showed that overcommitment (but not general anger) moderated the effect of ERI on driving anger; ERI has a greater influence on increasing driving anger in individuals with high overcommitment at work. The results have considerable implications for the safety and emotional health of individuals who perceive an imbalance between their efforts and rewards at work, and overcommitted individuals may be at greater risk. The wider implications of the relationship between work stress, emotional well-being and driving anger in employees, along with the potential of driver education interventions, are discussed as public health issues. PMID- 17920854 TI - Success on a practical driver's license test with and without the presence of another testee. AB - The present study examined whether performance in practical driving tests differed when individuals were tested with and without the presence of another testee. In addition, this study examined whether such differences were dependent on the sex of the tested person, on that of the observer testee, and on the order of tests. It was found that for both males and females, a greater proportion of drivers who were tested alone passed the test as compared to drivers who were tested in pairs. Additionally, proportionately more males than females passed the driving test. Only for males was performance also dependent on the gender of the other tested individual. A greater proportion of males who were tested with an accompanying female passed the test than males who were tested with an accompanying male. These results are discussed in relation to social facilitation, gender differences in driving performance, and differences in testing procedures. PMID- 17920855 TI - Characterization of food waste and bulking agents for composting. AB - The characterization of food waste (FW) and locally available bulking agents (BA) are a prerequisite to optimizing compost recipes. This study measured the variation in FW characteristics (pH, dry matter (DM), carbon (C), wet bulk density and Total Kjeldahl Nitrogen (TKN)) produced by a restaurant and a community kitchen in downtown Montreal, Canada from May to August 2004. The project also measured the mass of FW produced by another restaurant and a group of 20-48 households, from June to August 2004. Locally available BA (hay, straw, pine wood shavings, cardboard, left over cattle feed and wheat residue pellets) were also characterized to formulate composting recipes based on the FW characteristics observed during a period representative of winter and summer conditions. Residential and restaurant FW characteristics varied significantly over the summer months, although the mass produced remained constant at 0.61 and 0.56 kg capita(-1)day(-1), respectively. In addition, the number of customers served by the restaurant increased by nearly 50% from June to August. The BA with the highest moisture adsorption capacity was found to be the wheat residue pellets, followed by chopped straw. Wheat residue pellets, chopped hay and left over cattle feed all presented a balanced C/N ratio. Wheat residue pellets and wheat straw, chopped hay and cardboard demonstrated neutral pH values. Based on the variable FW characteristics and monthly production rates, the formulation of recipes indicates that compost facilities must be flexible enough to handle seasonal variations of as much as 50% by volume. PMID- 17920856 TI - Source separation of household waste: a case study in China. AB - A pilot program concerning source separation of household waste was launched in Hangzhou, capital city of Zhejiang province, China. Detailed investigations on the composition and properties of household waste in the experimental communities revealed that high water content and high percentage of food waste are the main limiting factors in the recovery of recyclables, especially paper from household waste, and the main contributors to the high cost and low efficiency of waste disposal. On the basis of the investigation, a novel source separation method, according to which household waste was classified as food waste, dry waste and harmful waste, was proposed and performed in four selected communities. In addition, a corresponding household waste management system that involves all stakeholders, a recovery system and a mechanical dehydration system for food waste were constituted to promote source separation activity. Performances and the questionnaire survey results showed that the active support and investment of a real estate company and a community residential committee play important roles in enhancing public participation and awareness of the importance of waste source separation. In comparison with the conventional mixed collection and transportation system of household waste, the established source separation and management system is cost-effective. It could be extended to the entire city and used by other cities in China as a source of reference. PMID- 17920857 TI - Methods for household waste composition studies. AB - The question is how to conduct household waste composition studies. The review is divided into three parts: overview of known methods, sampling theory, and the waste components. Twenty methods are listed and commented on. There is no adopted, working international standard. In accordance with Pierre Gy's Theory of Sampling, the seven types of sampling errors, when collecting and splitting solid samples, are described and commented on in relation to sampling of household solid waste. It is concluded that the most crucial choices in household waste composition studies are: to divide the investigation into relevant number and types of strata; to decide the required sample size and number of samples; to choose the sampling location, i.e., sampling at household level or sampling from loads of waste collection vehicles; and to choose the type and number of waste component categories to be investigated. Various classifications of household waste components used in composition studies are listed and discussed. Difficulties and weaknesses of the reviewed methods are discussed and concluded in suggested questions for further research. PMID- 17920858 TI - Epigenetics of antigen-receptor gene assembly. AB - The antigen receptor genes are organized into distinct DNA elements that encode the variable (V), diversity (D) and joining (J) regions. It is now well established that the rearrangement of antigen receptor genes is regulated by developmental-specific modulation of chromatin structure. Further studies involving statistical mechanics should provide physical insight into the physical mechanisms that underlie the association of antigen receptor gene segments. PMID- 17920859 TI - Stress responses of bacteria. AB - Bacteria, irrespective of natural habitat, are exposed to constant fluctuations in their growth conditions. Consequently they have developed sophisticated responses, modulated by the re-modelling of protein complexes and by phosphorylation-dependent signal transduction systems, to adapt to and to survive a variety of insults. Ultimately these signalling systems affect transcriptional regulons either by activating an alternative sigma factor subunit of RNA polymerase, for example, sigma E (sigma(E)) of Escherichia coli and sigma B (sigma(B)) and sigma F (sigma(F)) in Bacillus subtilis or by activating DNA binding two-component response regulators. Recent structure determinations, and systems biology analysis of key regulators in well-characterised stress responsive pathways, illustrate conserved and novel mechanisms in these representative model bacteria. PMID- 17920860 TI - Imaging in the investigation of paraneoplastic syndromes. AB - Paraneoplastic syndromes are a heterogeneous group of disease presentations caused by underlying tumours. As they are non-metastatic in nature an intensive diagnostic evaluation is warranted to identify potentially curable lesions. The selection of the appropriate method of imaging is important in these cases, especially when history and physical examination are unrevealing. In this review the important paraneoplastic syndromes and underlying malignancies are discussed along with relevant imaging strategies. PMID- 17920862 TI - MammoGrid--a prototype distributed mammographic database for Europe. AB - This paper describes the prototype for a Europe-wide distributed database of mammograms entitled MammoGrid, which was developed as part of an EU-funded project. The MammoGrid database appears to the user to be a single database, but the mammograms that comprise it are in fact retained and curated in the centres that generated them. Linked to each image is a potentially large and expandable set of patient information, known as metadata. Transmission of mammograms and metadata is secure, and a data acquisition system has been developed to upload and download mammograms from the distributed database, and then annotate them, rewriting the annotations to the database. The user can be anywhere in the world, but access rights can be applied. The paper aims to raise awareness among radiologists of the potential of emerging "grid" technology ("the second generation Internet"). PMID- 17920861 TI - MRI of overuse injury in elite athletes. AB - Overuse injuries are a common finding in elite athletes. Magnetic resonance imaging (MRI) is the optimal method for the diagnosis of overuse injury in athletes of all levels. We present a review of common and important overuse injuries occurring in elite athletes. A systematic approach based on the functional anatomic units - tendons, bones and joints - may assist in diagnosis of these injuries. PMID- 17920863 TI - A comparison of some anthropometric parameters between an Italian and a UK population: "proof of principle" of a European project using MammoGrid. AB - AIM: To demonstrate the use of grid technology to produce a database of mammograms and supporting patient data, specifically using breast density as a biomarker of risk for breast cancer, for epidemiological purposes. METHOD: The cohort comprised 1737 women from the UK and Italy, aged 28-87 years, mean 54.7 years, who underwent mammography after giving consent to the use of their data in the project. Information regarding height, weight, and exposure data (mAs and kV) was recorded. The computer program Generate-SMF was applied to all films in the database to measure breast volume, dense breast volume, and thereby percentage density. Visual readings of density using a six-category classification system were also available for 596 women. RESULTS: The UK and Italian participants were similar in height, but the UK women were significantly heavier with a slightly higher body mass index (BMI), despite being younger. Both absolute and percentage breast density were significantly higher in the Udine cohort. Images from the medio-lateral projection (MLO) give a significantly lower percentage density than cranio-caudal (CC) images (p<0.0001). Total breast volume is negatively associated with percentage density, as are BMI and age (p<0.0001 for all), although 80% of the variability in percentage density remains unexplained. CONCLUSION: The study offers proof of principle that confederated databases generated using Grid technology provide a useful and adaptable environment for large quantities of image, numerical, and qualitative data suitable for epidemiological research using the example of mammographic density as a biomarker of risk for breast cancer. PMID- 17920864 TI - Fluoroscopically guided percutaneous jejunostomy: outcomes in 25 consecutive patients. AB - AIM: To assess the feasibility and safety of fluoroscopically guided percutaneous jejunostomy. MATERIAL AND METHODS: Between May 1999 and August 2006 percutaneous jejunostomy was attempted in 25 patients. A 5 F vascular catheter (n=20) or a 7.5 F multifunctional coil catheter (n=5) was used to insufflate the jejunum. The distended jejunum was punctured using a 17 G needle (n=19) or a 21 G Chiba needle (n=6) with the inserted catheter as a target. A 12 or 14 F loop feeding tube was inserted after serial dilations. The technical success, complications, 30-day mortality, and in-dwelling period of the feeding tube placement were evaluated. RESULTS: The technical success rate was 92% (23/25). Technical failures (n=2) resulted from the inability to insufflate the jejunum secondary to failure to pass the catheter through a malignant stricture at the oesophagojejunostomy site and thus subsequent puncture of the undistended jejunum failed, or failure to introduce the Neff catheter into the jejunum. Pericatheter leakage with pneumoperitoneum was a complication in three patients (12%) and was treated conservatively. The 30-day mortality was 13% (3/23); however, there was no evidence that these deaths were attributed to the procedure. Except for four patients who were lost to follow-up and two failed cases, 15 of the 19 jejunostomy catheters were removed because of patient death (n=12) or completion of treatment (n=3), with a mean and median in-dwelling period of 231 and 87 days, respectively. CONCLUSIONS: Fluoroscopically guided percutaneous jejunostomy is a feasible procedure with a high technical success and a low complication rate. In addition to a 17 G needle, a 21 G needle can safely be used to puncture the jejunum. PMID- 17920866 TI - Visual search behaviour in skeletal radiographs: a cross-specialty study. AB - AIM: To determine whether experience improves the consistency of visual search behaviour in fracture identification in plain radiographs, and the effect of specialization. MATERIAL AND METHODS: Twenty-five observers consisting of consultant radiologists, consultant orthopaedic surgeons, orthopaedic specialist registrars, orthopaedic senior house officers, and accident and emergency senior house officers examined 33 skeletal radiographs (shoulder, hand, and knee). Eye movement data were collected using a Tobii 1750 eye tracker with levels of diagnostic confidence collected simultaneously. Kullback-Leibler (KL) divergence and Gaussian mixture model fitting of fixation distance-to-fracture were used to calculate the consistency and the relationship between discovery and reflective visual search phases among different observer groups. RESULTS: Total time spent studying the radiograph was not significantly different between the groups. However, the expert groups had a higher number of true positives (p<0.001) with less dwell time on the fracture site (p<0.001) and smaller KL distance (r=0.062, p<0.001) between trials. The Gaussian mixture model revealed smaller mean squared error in the expert groups in hand radiographs (r=0.162, p=0.07); however, the reverse was true in shoulder radiographs (r=-0.287, p<0.001). The relative duration of the reflective phase decreases as the confidence level increased (r=0.266, p=0.074). CONCLUSIONS: Expert search behaviour exhibited higher accuracy and consistency whilst using less time fixating on fracture sites. This strategy conforms to the discovery and reflective phases of the global-focal model, where the reflective search may be implicated in the cross-referencing and conspicuity of the target, as well as the level of decision-making process involved. The effect of specialization appears to change the search strategy more than the effect of the length of training. PMID- 17920867 TI - Magnetic resonance imaging of facial muscles. AB - Facial and tongue muscles are commonly involved in patients with neuromuscular disorders. However, these muscles are not as easily accessible for biopsy and pathological examination as limb muscles. We have previously investigated myasthenia gravis patients with MuSK antibodies for facial and tongue muscle atrophy using different magnetic resonance imaging sequences, including ultrashort echo time techniques and image analysis tools that allowed us to obtain quantitative assessments of facial muscles. This imaging study had shown that facial muscle measurement is possible and that useful information can be obtained using a quantitative approach. In this paper we aim to review in detail the methods that we applied to our study, to enable clinicians to study these muscles within the domain of neuromuscular disease, oncological or head and neck specialties. Quantitative assessment of the facial musculature may be of value in improving the understanding of pathological processes occurring within facial muscles in certain neuromuscular disorders. PMID- 17920868 TI - The value of hyoscine butylbromide in pelvic MRI. AB - AIM: To evaluate the effect of hyoscine butylbromide (HBB) on image quality and lesion and organ visualization in pelvic magnetic resonance imaging (MRI) MATERIALS AND METHODS: A prospective, ethically approved study was undertaken of 47 patients attending for pelvic MRI at a cancer centre. T2-weighted transverse and sagittal sequences were performed before and after intravenous injection of 20 mg HBB. Three radiologists independently scored anonymized image series for overall image quality, visualization of pelvic lesions and visualization of individual pelvic organs. Statistical analysis was performed to assess improvements in radiologists' scores post-HBB administration. Radiologists also assessed pre-HBB administration T1-weighted images for degree of bowel peristalsis to determine whether this could predict improvement in post-HBB T2 weighted image scores. Side effects of HBB were recorded using a patient questionnaire. RESULTS: Radiologists' scores for image quality and lesion visualization were significantly higher on the post-HBB administration T2 weighted series (p<0.0005). Scores for the visualization of the bladder, rectum, pelvic bowel, prostate, and seminal vesicles (all p<0.0005), cervix (p=0.019) and vagina (p=0.0001) were also significantly higher post-HBB administration. Scores for the degree of peristalsis on T1-weighted images were not related to improvement in image quality or lesion visualization on T2-weighted images post HBB administration. Side effects of HBB were mild and self-limiting. CONCLUSION: Intravenous HBB administration improves image quality and lesion visualization in oncological pelvic MRI and is recommended for routine use. PMID- 17920869 TI - CT-guided sacroplasty for the treatment of sacral insufficiency fractures. AB - AIM: To describe the clinical presentation, procedure and outcome in patients treated with computed tomography (CT)-guided sacroplasty as a treatment for sacral insufficiency fractures. MATERIALS AND METHODS: Three patients (mean age 80 years, range 75-87 years) were treated with CT-guided sacroplasty. The mean pre-procedure visual analogue score (VAS) for pain was 8 (range 7-9) with a mean symptom duration of 8 months (range 2.5-18). The procedure was performed under CT guidance with needles being placed along the fracture lines from a posterior approach. Polymethylmethacrylate (PMMA) cement was introduced in 0.2 ml aliquots after cement temperature reduction. Cement injection was monitored by four section block axial acquisition to assess potential cement migration. RESULTS: All three procedures were performed without significant complication. One patient developed a tiny asymptomatic cement leak into the S1 foramen. The mean volume of cement injected into a unilateral sacral fracture was 4 ml. All patients tolerated the procedure well under intravenous sedation. The mean VAS score post procedure was 2. Continued symptomatic relief was seen at 6 weeks and 3 months. CONCLUSION: CT-guided sacroplasty represents an alternative treatment for sacral insufficiency fractures that are resistant to conservative treatment. The symptomatic relief the procedure produces seems to be excellent both in this small series and in described cases in the literature. PMID- 17920871 TI - Pseudoaneurysm of the renal artery following partial nephrectomy: imaging findings and coil embolization. AB - AIM: To present the imaging findings of five patients with renal artery pseudoaneurysm (RAP) after partial nephrectomy. METHODS: Five patients (four men and one woman) with RAP as a complication of partial nephrectomy were studied. The diagnosis of RAP was established using contrast-enhanced computed tomography (CT) in three patients and renal angiography in two patients. In two cases, the diagnosis was evident on ultrasound with colour Doppler. RESULTS: The indication for partial nephrectomy (open approach in four patients and laparoscopic in one patient) was a space-occupying lesion, which proved to be a renal cell carcinoma. All patients presented with macroscopic haematuria, 1-21 days (mean 12.2 days) after surgery. In three of patients the definitive diagnostic imaging method was contrast-enhanced CT. The arterial phase of CT showed a well-circumscribed dense collection of contrast material located within the renal parenchyma. In two other patients the initial and conclusive diagnostic imaging method was renal angiography. All patients underwent selective renal angiography with therapeutic coil embolization. The procedure failed in one patient, which necessitated nephrectomy. CONCLUSIONS: Pseudoaneurysm of the renal artery should be considered in patients presenting with macrohaematuria after nephron-sparing surgery. The diagnosis can be established using contrast-enhanced CT, ultrasound with colour Doppler, or angiography. Renal angiography with selective embolization is a safe and efficacious technique for managing the condition. PMID- 17920873 TI - The "two-tone" testis: spectrum of ultrasound appearances. AB - AIM: To determine the prevalence and variation in appearance of the "two-tone" testes artefact. METHODS AND MATERIALS: Patients attending for a testicular ultrasound, without an intra-testicular lesion, were included. A single operator performed examinations using a 15L8w linear array transducer. When an artefact resembling the "two-tone" testes appearance was seen, the images were recorded prospectively. Two independent observers reviewed the images and identified patterns of the two-tone testis as: partial posterior shadowing from an incompletely identified trans-testicular artery; partial posterior shadowing from a completely identified trans-testicular artery; total posterior shadowing from a completely identified trans-testicular artery. RESULTS: Over a 44-month period 1038 patients were examined. The median age of patients was 32.5 years (range 19 46 years). A total of 17 two-tone testes artefacts of varying appearances were identified in 16 patients, occurring in 1.6% of the selected population. There were six partial posterior shadowing from an incompletely identified trans testicular artery (0.6%); five partial posterior shadowing from a completely identified trans-testicular artery (0.5%); and five total posterior shadowing from a completely identified trans-testicular artery (0.5%) artefacts present. There was no relationship between the two-tone artefact and clinical symptoms or other ultrasound findings. CONCLUSION: The two-tone testis artefact is a rare finding and has a variable appearance according to the extent of acoustic shadowing produced. Careful examination of the testis and operator experience is required when the artefact is identified in order that underlying pathological disease is not overlooked. PMID- 17920872 TI - Classification of temporal bone pneumatization based on sigmoid sinus using computed tomography. AB - AIM: To analyse several reference structures using axial computed tomography (CT) imaging of the temporal bone, which may reflect pneumatization of the entire temporal bone by statistical correlation to the actual volume of the temporal bone measured using three-dimensional reconstruction. MATERIALS AND METHODS: One hundred and sixteen temporal bones were studied, comprising 48 with normal findings and 68 sides showing chronic otitis media or temporal bone fracture. After measuring the volume of temporal bone air cells by the volume rendering technique using three-dimensional reconstruction images, classification of temporal bone pneumatization was performed using various reference structures on axial images to determine whether significant differences in the volume of temporal bone air cells could be found between the groups. RESULTS: When the sigmoid sinus at the level of the malleoincudal complex was used in the classification, there were statistically significant differences between the groups that correlated with the entire volume of the temporal bone. Grouping based on the labyrinth and the ascending carotid artery showed insignificant differences in volume. Furthermore, there was no significant correlation between the cross-sectional area of the antrum and the entire volume of the temporal bone. CONCLUSION: The degree of pneumatization of temporal bone can be estimated easily by the evaluation of the air cells around the sigmoid sinus on axial CT images. PMID- 17920874 TI - Fibrosing mediastinitis associated with Behcet's disease: CT findings. PMID- 17920876 TI - Enhancing the radiology product: the value of voice-recognition technology. PMID- 17920875 TI - Is eLD an effective way of revising for FRCR 2A? PMID- 17920877 TI - Analysis of trend in radiology subspecialties in the UK. PMID- 17920878 TI - Wheat bran biodegradation by Pleurotus ostreatus: a solid-state carbon-13 NMR study. AB - Solid-state (13)C nuclear magnetic resonance (NMR) and elemental analysis techniques were used to monitor the degradation of wheat bran by the white-rot fungus Pleurotus ostreatus during a 62-day cultivation period. The weight loss and in vitro organic matter digestibility of the substrate were also evaluated after fungal treatment. The (13)C NMR spectra of degraded wheat bran samples showed a lower content in carbohydrates and a higher content in aliphatic and carboxylic groups than the untreated control sample. In parallel, changes in the wheat bran elemental composition evidenced a decrease in carbon content and a concomitant increase in nitrogen and oxygen content during mycelium growth. These results clearly indicate the occurrence of progressive changes in the composition of wheat bran during fungal treatment and are interpreted in terms of preferential degradation of amorphous vs. crystalline polysaccharides by the fungal mycelium and accumulation of proteins in the substrate. At the end of the cultivation period, the treated samples experienced an average weight loss of 20% and an increase in organic matter digestibility of 17%. PMID- 17920879 TI - Comparison of in vitro Cr(VI) reduction by CFEs of chromate resistant bacteria isolated from chromate contaminated soil. AB - Chromate resistant and reducing strains were isolated from chromium contaminated soil and identified as Bacillus sp. (KCH2 and KCH3), Leucobacter sp. (KCH4) and Exiguobacterium sp. (KCH5). KCH3 and KCH4 showed higher Cr(VI) tolerance (2 mM) and Cr(VI) reduction (1.5 mM) than KCH5 (1.5 mM and 0.75 mM, respectively). Cr(VI) reduction by CFEs of KCH3 and KCH4 showed NAD(P)H dependence, optimum activity at pH 5.5, low K(m) (45-55 microM) and substrate inhibition by Cr(VI) (>75 microM), whereas that of KCH5 showed NADH dependence, pH optimum at 6.0, high K(m) (200 microM) and no inhibition by Cr(VI). Cr(VI) reduction was optimum at 35 degrees C for CFEs of KCH3 and KCH5 and 30 degrees C for that of KCH3. Cr(VI) reduction by CFEs of all the strains were inhibited by Hg(2+) and enhanced by Cu(2+). Activity enhancement by Cu(2+) was more predominant (290%) for KCH4. The characterization of Cr(VI) reduction by CFEs of chromate resistant isolates of different genera is useful for development of Cr(VI) bioremediation. PMID- 17920880 TI - Studies on the antioxidant activity of the essential oil and methanol extract of Marrubium globosum subsp. globosum (lamiaceae) by three different chemical assays. AB - This study is designed to examine the chemical composition and in vitro antioxidant activity of the essential oil and sub-fractions of the methanol extract of Marrubium globosum subsp. globosum. The GC and GC-MS analysis of the essential oil were resulted in the determination of 84 components representing 88.2% of the oil. The major constituents of the oil were spathulenol (15.8%), beta-caryophyllene (9.0%), caryophyllene oxide (7.9%), germacrene D (6.5%), and bicyclogermacrene (3.1%). Antioxidant activities of the samples were determined by three different test systems namely DPPH, beta-carotene/linoleic acid and reducing power assay. In DPPH system, the weakest radical scavenging activity was exhibited by the essential oil (1203.38+/-7.18 microg ml(-1)). Antioxidant activity of the polar sub-fraction of methanol extract was superior to the all samples tested with an EC(50) value of 157.26+/-1.12 microg ml(-1). In the second case, the inhibition capacity (%) of the polar sub-fraction of methanol extract (97.39%+/-0.84) was found the strongest one, which is almost equal to the inhibition capacity of positive control BHT (97.44%+/-0.74). In the case of reducing power assay, a similar activity pattern was observed as given in the first two systems. Polar sub-fraction was the strongest radical reducer when compared with the non-polar one, with an EC(50) value of 625.63+/-1.02 microg ml( 1). The amount of the total phenolics was highest in polar sub-fraction (25.60+/ 0.74 microg/mg). A positive correlation was observed between the antioxidant activity potential and total phenolic level of the extracts. On the other hand, total flavonoid content was found equal for the both sub-fractions. PMID- 17920881 TI - Kinetic characteristics of n-butyl alcohol and iso-butyl alcohol in a composite bead air biofilter. AB - Kinetic characteristics of n-butyl alcohol and iso-butyl alcohol in a composite bead biofilter were investigated. The microbial growth rate of n-butyl alcohol was greater than that of iso-butyl alcohol in the average inlet concentration range of 50-300 ppm. The microbial growth rate was inhibited at higher inlet concentration, and the inhibitive effect in the concentration range of 50-150 ppm was more pronounced than that in the concentration range of 150-300 ppm. The degree of inhibitive effect for n-butyl alcohol was more sensitive than that for iso-butyl alcohol in the concentration range of 50-150 ppm. The zero-order kinetic with the diffusion rate limitation could be regarded as the most adequate biochemical reaction model. The biodegradation rate of n-butyl alcohol was greater than that of iso-butyl alcohol in the average inlet concentration range of 50-300 ppm. The biochemical reaction rate was also inhibited at higher inlet concentration, and the inhibitive effect for iso-butyl alcohol was more pronounced than that for n-butyl alcohol. The factor of the chemical structure of compound was more predominant in the microbial growth and biochemical reaction processes. The maximum elimination capacity of n-butyl alcohol and iso-butyl alcohol were 55.7 and 34.8 g C h(-1)m(-3) bed volume, respectively. The compound with no side group in the main chain would be easier biodegraded by the microbial. PMID- 17920882 TI - Kinetics and thermodynamics of basic dye sorption on phosphoric acid esterifying soybean hull with solid phase preparation technique. AB - In this paper, the solid phase preparation method of a cationic sorbent, which bears hydroxyl groups of phosphoric acid derived from esterified soybean hull (ESH), was reported. The sorption kinetics and thermodynamics of two basic dyes, acridine orange (AO) and malachite green (MG), from aqueous solution onto ESH were investigated with a batch system. The isothermal data of dye sorptions followed the Langmuir model better than the Freundlich model. The maximum sorption capacity (Q(m)) of ESH for AO and MG was 238.1 mg/g and 178.57 mg/g, respectively. The dye sorption processes could be described by the pseudo-second order kinetic model. The thermodynamic study indicated that the dye sorptions were spontaneous and exothermic. Lower temperatures were favorable for the sorption processes. PMID- 17920883 TI - Optimization of methane fermentation from effluent of bio-hydrogen fermentation process using response surface methodology. AB - The individual effects and interactive effects of substrate concentration, ratio of inoculum to substrate, Ca(2+) concentration on the methane yield from the effluent of bio-hydrogen fermentation of food waste were investigated in this study. A central composite design (CCD) and response methodology (RSM) were employed in designing the experiments, in order to determine the optimum conditions for methane fermentation. The experiment results showed that the effects of substrate concentration, ratio of inoculum to substrate, Ca(2+) concentration were statistically significant at 5% level. The interactive effect of substrate concentration and ratio of inoculum to substrate was significant, however the interactive effect of substrate concentration and Ca(2+) concentration, ratio of inoculum to substrate and Ca(2+) concentration were found to be insignificant at 5% level. A maximum yield of 565.76 ml CH(4)/g VS(added) was estimated under the optimum conditions for substrate concentration 7.77 g of VS/l, inoculum to substrate ratio of 2.81 and calcium concentration of 380.82 mg/l. Verification experiment of the estimated optimum conditions confirmed that the RSM was useful for optimizing the methane yield from effluent of bio-hydrogen fermentation of food waste. PMID- 17920884 TI - New 'chemical probes' to examine the role of the hFPRL1 (or ALXR) receptor in inflammation. AB - We report the development of the novel N-substituted benzimidazole 11 as a potent and selective human formyl peptide receptor-like 1 (hFPRL1) agonist. This compound and its less active enantiomer 12 were identified as useful tools for studying receptor function in vitro. PMID- 17920885 TI - Synthesis of novel phytosphingosine derivatives and their preliminary biological evaluation for enhancing radiation therapy. AB - Eight d-ribo-phytosphingosine derivatives were synthesized from d-ribo phytosphingosine and diverse acyl chlorides with N,N-diisopropylethylamine in tetrahydrofuran for 1h at room temperature. Effect of these compounds on IR induced cell death was evaluated on blood cancer cells (Jurkat). Among these, 3d showed the highest enhancement of radiosensitizing effect. PMID- 17920886 TI - Kinetic mechanisms to alter walking speed. AB - A mechanism to modulate speed during human walking has not yet been proposed in the literature, even though changing walking speed is likely a necessary attribute of everyday ambulation. To understand how joint kinetics modulate walking speed 12 normal adults walked Fast (1.4m/s), Slow (1.0m/s), Accel (1.0 1.4m/s) and Decel (1.4-1.0m/s) trials while full body 3D kinematics and kinetics were collected. Reduced sagittal ankle plantarflexor moments were observed in Accel trials during early single limb stance (p<0.001) and increased sagittal plantarflexor moments were seen in Decel trials during early single limb stance (p<0.001) compared to steady speed walking. Modulating the sagittal ankle moment altered the center of pressure location and either attenuated (Accel) or accentuated (Decel) the early stance braking impulse to accelerate or decelerate the center of mass. The onset of walking speed changes occurred at approximately 15% of the gait cycle and did not support the concept of "controlled falling". Sagittal ankle push-off power appears a consequence of increased walking speed, but not the causative factor to increase walking speed. PMID- 17920887 TI - Introduction of a new mobile-bearing total knee prosthesis: minimum three year follow-up of an RCT comparing it with a fixed-bearing device. AB - As part of the step-wise validation of a new prosthesis (TMK), we previously published the 1 year results of a randomised controlled trial in patients undergoing bilateral knee replacement [Price A., Rees J., Beard D., Juszczak E. et al. A mobile-bearing total knee prosthesis compared with a fixed-bearing prosthesis. JBJS B 2003;85-B-1:62-7.]. Forty patients had the new mobile-bearing prosthesis implanted in one knee and an established fixed-bearing device in the other (AGC). We now report the 3 year status of these patients and, in addition, review a separate multi-centre cohort of 172 patients who had undergone unilateral arthroplasty with the TMK. No significant differences were found in outcome (American Knee Society Score and Oxford Knee Score) between the two prostheses. The greater incidence of "clicking" in the mobile-bearing knee, reported in the previous review, persisted (TMK=48%, AGC=30%). The presence of this mechanical noise was found to have no relationship with outcome in either of the prostheses. The unilateral cohort study showed an acceptable complication rate for the new prosthesis, although some patients reported subjective instability. The method of controlled introduction of the TMK, of which this constitutes a further step, has allowed us to assess the significance of a reported problem (clicking) and to provide scientific data from which other surgeons can decide about use of the implant. PMID- 17920888 TI - Pyrene-perylene as a FRET pair coupled to the N2'-functionality of 2'-amino-LNA. AB - Detection of nucleic acid hybridization via fluorescence resonance energy transfer (FRET) using pyren-1-ylmethyl and perylen-3-ylmethyl N2'-functionalized 2'-amino-LNA nucleosides incorporated into oligonucleotides exhibited a clear distance dependence of the FRET efficiency, ranging from below 10% when the fluorophores were approximately 40A apart to approximately 90% when the fluorophores were in close proximity. PMID- 17920890 TI - Practical aspects of delivering hereditary cancer risk counseling. AB - The primary element involved in the practical delivery of hereditary cancer risk counseling is the development of a cancer genetics service. This service should involve a multidisciplinary core of specialists, aim to identify at-risk individuals and their families, and provide comprehensive genetic counseling with the option of genetic testing. This article examines the development of a cancer genetics service beginning with the fundamentals of patient ascertainment, location, staffing, and billing. It also reviews the elements involved in the provision of genetic counseling services, including formalized risk assessment and risk communication. Finally, we review novel modes of delivery of genetic counseling including the use of telephone counseling and telemedicine as a means of expanding the scope of this service and extending its reach to individuals who might otherwise not have access to a risk assessment clinic. Overall, this presentation can serve as a resource for identifying the practical aspects involved in the delivery of hereditary cancer risk counseling and their subsequent integration into the workings of a proficient cancer genetics service able to identify and serve at-risk patients and their family members. PMID- 17920891 TI - Clinically relevant biology of hereditary breast cancer. AB - The identification of the two major breast cancer susceptibility genes BRCA1 and BRCA2 has led to an explosion of work covering the biology and epidemiology of these genes. Clinically, much is known about the risks associated with mutations in these genes, but less is certain with regards to their impact on treatment. This review will touch upon several important aspects of the biology of BRCA1 and BRCA2, but comments regarding these areas will be restricted to those most relevant to the clinician. The three main sections are molecular pathology, DNA repair, and finally other functions of the BRCA1 and BRCA2 proteins. PMID- 17920893 TI - Screening for hereditary breast cancer. AB - Most women at risk for hereditary breast cancer opt for intensive breast screening rather than risk-reducing mastectomy. For this to be a rational choice, the vast majority of tumors must be detected either while still in situ or at a very early stage of invasion. Annual screening mammography has low sensitivity in this population, in part due to the greater breast density of younger women, resulting in cancers being detected at a suboptimal stage. In six prospective comparative studies, the addition of annual contrast-enhanced magnetic resonance imaging (MRI) of the breast to mammography demonstrated greater than 90% sensitivity, more than twice that of mammography alone. In those studies that included ultrasound and clinical breast examination, additional cancers were rarely detected by these modalities. False positive rates were higher with the addition of MRI, but specificity improved on successive rounds of screening. Although long-term survival data are still lacking, there is mounting evidence that the addition of screening MRI to mammography detects hereditary breast cancers at an earlier stage and is thus estimated to be cost-effective, at least for women with BRCA mutations. This review will examine the literature and current screening recommendations. PMID- 17920892 TI - Treatment of hereditary breast cancer. AB - Mutations in BRCA1 and BRCA2 are well-established causes of hereditary breast cancer. As genetic testing becomes more widespread, increasing numbers of women are known to have mutations at or shortly after their breast cancer diagnosis. Current evidence is insufficient to mandate different local or systemic treatment based upon the presence of a germline mutation. The well-documented increased risk of contralateral second primary breast cancer and possibly of late ipsilateral second primary breast cancers may influence patient decision-making with regard to breast-conserving treatment. PMID- 17920894 TI - The prevention of hereditary breast cancer. AB - In the past 10 years our knowledge of hereditary breast cancer, and specifically the breast cancer susceptibility genes BRCA1 and BRCA2, has dramatically increased, as has our understanding of prevention options for high-risk women. Effective strategies to reduce the risk of breast cancer include prophylactic mastectomy, prophylactic oophorectomy, and chemoprevention. In this article we review the current evidence regarding the efficacy of each of these management strategies. PMID- 17920895 TI - Screening and prevention of hereditary gynecologic cancers. AB - Endometrial and ovarian cancer are the fourth and fifth most common malignancies in women, with approximately 40,000 new endometrial and 25,000 new ovarian cancers expected to be diagnosed in the Unites States this year. While the majority of these cancers will occur in the absence of a family history, approximately 5% of endometrial cancers and 10% of ovarian cancers will be the result of inherited defects in high-penetrance cancer susceptibility genes. With the identification and subsequent availability of clinical genetic testing for mutations in the genes associated with hereditary breast-ovarian cancer and the Lynch/hereditary nonpolyposis colorectal cancer (HNPCC) syndrome, targeted risk reduction using intensive screening, chemoprevention, and prophylactic surgery has become possible for women at inherited risk of gynecologic malignancies. We review the options for gynecologic cancer risk reduction in women with an inherited mutation in BRCA1, BRCA2, or one of the mismatch repair (MMR) genes associated with Lynch/HNPCC syndrome. Additionally, we outline ongoing questions and areas for future research. PMID- 17920896 TI - Strategies for identifying hereditary nonpolyposis colon cancer. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is the term given to a predisposition syndrome caused by inherited mutations in one of at least five DNA mismatch repair (MMR) genes. Dominant mutations in these genes predispose individuals to a range of cancers in addition to the most frequent, colorectal cancer. Endometrial cancer is the most notable additional malignancy, followed by ovarian, gastric, upper urethelial, and biliary cancers, and gliomas. Recognition of HNPCC is important so that targeted screening can be effected that will reduce the incidence of the main cancers. While such clinical criteria as Amsterdam and modified Amsterdam are reasonably specific, they lack sensitivity. Thus, tumor related features have been used to improve sensitivity for identifying patients who can be selected for the relatively expensive direct mutation analysis of the various genes. Microsatellite instability (MSI) and loss of antibody staining for the proteins have been widely vaunted but have their own drawbacks. No one approach has received universal acceptance, and therefore adoption of one of perhaps three strategies, including clinical- and laboratory-based approaches, is still appropriate until an easier, quicker, and cheaper approach can be developed. PMID- 17920898 TI - Familial colorectal cancer syndrome X. AB - In recent decades there have been significant advances in our understanding of the genes that underlie hereditary susceptibility to colorectal cancer (CRC). In 2007 it is well established that mutations in DNA repair genes (MLH1, MSH2, MSH6, MYH) and Wnt pathway signal transduction inhibitors (APC) underlie a significant percentage of hereditary CRC susceptibility. However, it also is clear that the known CRC susceptibility genes do not explain fully the inherited risk seen even in families meeting the revised Bethesda guidelines. Furthermore, the optimal medical management of these syndromes is still being defined. What underlies CRC susceptibility in these highly unusual families that do not have identifiable mutations in the known genes, often referred to as syndrome X? This review addresses this important question that is relevant to our current understanding of the management of individuals with hereditary predisposition to CRC. PMID- 17920897 TI - Recently identified colon cancer predispositions: MYH and MSH6 mutations. AB - Single-gene germline mutations conferring a high lifetime risk of colorectal cancer (CRC) account for up to 6% of all CRC cases. The most widely studied monogenic colorectal cancer syndromes include familial adenomatous polyposis (FAP) and Lynch syndrome. However, additional syndromes continue to be defined and new predisposition genes are continuing to be identified. Most recently, MYH associated polyposis (MAP) and an "atypical Lynch syndrome" related to the presence of MSH6 mutations have been linked to an increased risk of CRC. In this review, we summarize basic information related to these newly recognized gene mutations, including the accumulating data on the prevalence and penetrance of deleterious mutations, as well as the management options for identified carriers and their families. Recognizing these heritable syndromes is essential and predictive genetic testing will continue to transform the field of cancer risk assessment by offering the opportunity to focus on more precise risk management and cancer prevention. PMID- 17920899 TI - Cowden syndrome. AB - Cowden syndrome (CS), due to germline mutations of the PTEN tumor-suppressor gene, is an often overlooked cancer predisposition syndrome associated with an increased risk of breast, thyroid, and endometrial cancers, as well as benign manifestations. Germline PTEN mutations also are associated with syndromes that have not been historically connected to an increase in risk for malignancy. These disorders include Bannayan-Riley-Ruvalcaba syndrome (BRRS), Proteus syndrome (PS), and Proteus-like syndrome (PSL). These syndromes can be described under the umbrella of PTEN hamartoma tumor syndrome (PHTS). As one would expect in allelic disorders, there is broad phenotypic overlap in the PHTS; however, the syndromes are clinically distinct. As additional information is discovered about new syndromes of cancer predisposition and their concordant genes, oncologists and allied healthcare providers must maintain vigilance to appropriately identify, and screen, individuals at an increased risk. Although CS is the only PHTS with a clearly documented predisposition to malignancies, pending further data, for precautionary reasons all individuals with a germline PTEN mutation are recommended to follow the cancer surveillance recommendations for CS. PMID- 17920900 TI - Ethical and legal aspects of cancer genetic testing. AB - As a result of the increasing effectiveness of cancer screening and preventive interventions, ethical issues, as well as legal liabilities, are increasingly associated with cancer genetic testing. These issues include the possible "duty to warn" relatives of inherited cancer risk, the appropriateness of testing of children and embryos, equity of access to genetics services, and potential harms of testing including the risk of genetic discrimination. An approach to these and other ethical challenges will be presented, drawing not only on recent case law but also on a broader bioethical framework. PMID- 17920901 TI - Influence of cell preparation and target location on the behavioral recovery after striatal transplantation of fetal dopaminergic neurons in a primate model of Parkinson's disease. AB - Surgeries involving transplantation of fetal dopamine (DA) neurons into the caudate-putamen of patients with Parkinson's disease (PD) have been performed in various clinical trials to examine a potential restoration of motor function. The absence of studies in non-human primates to define the best transplantation protocols have lead to the use of a broad variety of techniques that potentially could have a major impact on the clinical outcome. The effects of using different cell and tissue preparation, and surgical targets, remain unknown. For this purpose, 20 St. Kitts African Green Monkeys (AFG) rendered parkinsonian by i.m. injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) were balanced into 4 groups and unilaterally grafted in the (a) caudate or (b) putamen with fetal ventral mesencephalic (VM) tissue as (c) solid pieces or as a (d) cell suspension. By 9 months post-transplantation all animals showed significant and similar behavioral improvement as determined by a UPDRS based PD scale. Postmortem analyses showed that VM transplants survived in all animals. They were located in both surgical target sites, producing a broad DA reinnervation of the targeted nuclei that could also extend to the non-grafted nucleus on the ipsilateral side. Although no differences between groups were found in survival of DA neurons or degree of DA reinnervation, there was a significant correlation between striatal reinnervation and behavioral recovery only in animals transplanted in the putamen surgical target. Additionally, there was in general a stronger glial reaction to solid grafts than to cell suspensions. These studies provide data for the optimal time course, cell preparation and surgical targets for systematic examinations of both potential benefits and side effects of dopamine neuron cell transplantation in primate models of PD. PMID- 17920903 TI - Adding harm reduction to tobacco control. PMID- 17920904 TI - Medicines for children: safety as an afterthought. PMID- 17920902 TI - The high affinity peripheral benzodiazepine receptor ligand DAA1106 binds to activated and infected brain macrophages in areas of synaptic degeneration: implications for PET imaging of neuroinflammation in lentiviral encephalitis. AB - HIV encephalitis (HIVE) is characterized by neurodegeneration mediated by toxins derived from infected and activated brain macrophages. Since the peripheral benzodiazepine receptor (PBR) is abundant on brain macrophages, we hypothesized that [(3)H]DAA1106, a new PBR ligand, can label infected and activated brain macrophages in HIVE. Using cell culture and postmortem brain tissues from HIVE and a macaque model of HIVE, we show that [(3)H]DAA1106 binds with high affinity to activated and infected macrophages in regions of synaptic damage. Further, binding affinity reflected by lower K(D) (dissociation constant) values and the B(max) (total number of binding sites) to K(D) ratios reflective of ligand binding potential was significantly higher with [(3)H]DAA1106 compared to the extensively characterized PBR ligand [(3)H](R)-PK11195. These data suggest that DAA1106 binds with high affinity to activated and infected brain macrophages and possesses binding characteristics beneficial for in vivo use in the detection and clinical monitoring of HIVE using positron emission tomography. PMID- 17920905 TI - Financing the fight against AIDS, tuberculosis, and malaria. PMID- 17920906 TI - A new standard of care for elderly patients with myeloma. PMID- 17920907 TI - Day or night blood pressures to predict cardiovascular events? PMID- 17920908 TI - Effect of zinc supplementation on child mortality. PMID- 17920910 TI - Andrew Dillon. PMID- 17920911 TI - Swedish snus for tobacco harm reduction. PMID- 17920913 TI - Swedish snus for tobacco harm reduction. PMID- 17920914 TI - Snus and the risk of cancer of the mouth, lung, and pancreas. PMID- 17920916 TI - Melphalan and prednisone plus thalidomide versus melphalan and prednisone alone or reduced-intensity autologous stem cell transplantation in elderly patients with multiple myeloma (IFM 99-06): a randomised trial. AB - BACKGROUND: In multiple myeloma, combination chemotherapy with melphalan plus prednisone is still regarded as the standard of care in elderly patients. We assessed whether the addition of thalidomide to this combination, or reduced intensity stem cell transplantation, would improve survival. METHODS: Between May 22, 2000, and Aug 8, 2005, 447 previously untreated patients with multiple myeloma, who were aged between 65 and 75 years, were randomly assigned to receive either melphalan and prednisone (MP; n=196), melphalan and prednisone plus thalidomide (MPT; n=125), or reduced-intensity stem cell transplantation using melphalan 100 mg/m2 (MEL100; n=126). The primary endpoint was overall survival. Analysis was by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00367185. FINDINGS: After a median follow-up of 51.5 months (IQR 34.4-63.2), median overall survival times were 33.2 months (13.8 54.8) for MP, 51.6 months (26.6-not reached) for MPT, and 38.3 months (13.0-61.6) for MEL100. The MPT regimen was associated with a significantly better overall survival than was the MP regimen (hazard ratio 0.59, 95% CI 0.46-0.81, p=0.0006) or MEL100 regimen (0.69, 0.49-0.96, p=0.027). No difference was seen for MEL100 versus MP (0.86, 0.65-1.15, p=0.32). INTERPRETATION: The results of our trial provide strong evidence to indicate that the use of thalidomide in combination with melphalan and prednisone should, at present, be the reference treatment for previously untreated elderly patients with multiple myeloma. PMID- 17920917 TI - Prognostic accuracy of day versus night ambulatory blood pressure: a cohort study. AB - BACKGROUND: Few studies have formally compared the predictive value of the blood pressure at night over and beyond the daytime value. We investigated the prognostic significance of the ambulatory blood pressure during night and day and of the night-to-day blood pressure ratio. METHODS: We did 24-h blood pressure monitoring in 7458 people (mean age 56.8 years [SD 13.9]) enrolled in prospective population studies in Denmark, Belgium, Japan, Sweden, Uruguay, and China. We calculated multivariate-adjusted hazard ratios for daytime and night-time blood pressure and the systolic night-to-day ratio, while adjusting for cohort and cardiovascular risk factors. FINDINGS: Median follow-up was 9.6 years (5th to 95th percentile 2.5-13.7). Adjusted for daytime blood pressure, night-time blood pressure predicted total (n=983; p<0.0001), cardiovascular (n=387; p<0.01), and non-cardiovascular (n=560; p<0.001) mortality. Conversely, adjusted for night time blood pressure, daytime blood pressure predicted only non-cardiovascular mortality (p<0.05), with lower blood pressure levels being associated with increased risk. Both daytime and night-time blood pressure consistently predicted all cardiovascular events (n=943; p<0.05) and stroke (n=420; p<0.01). Adjusted for night-time blood pressure, daytime blood pressure lost prognostic significance only for cardiac events (n=525; p> or =0.07). Adjusted for the 24-h blood pressure, night-to-day ratio predicted mortality, but not fatal combined with non-fatal events. Antihypertensive drug treatment removed the significant association between cardiovascular events and the daytime blood pressure. Participants with systolic night-to-day ratio value of 1 or more were older, at higher risk of death, and died at an older age than those whose night-to-day ratio was normal (> or =0.80 to <0.90). INTERPRETATION: In contrast to commonly held views, daytime blood pressure adjusted for night-time blood pressure predicts fatal combined with non-fatal cardiovascular events, except in treated patients, in whom antihypertensive drugs might reduce blood pressure during the day, but not at night. The increased mortality in patients with higher night-time than daytime blood pressure probably indicates reverse causality. Our findings support recording the ambulatory blood pressure during the whole day. PMID- 17920919 TI - How shingles can be beached. PMID- 17920918 TI - Effect of daily zinc supplementation on child mortality in southern Nepal: a community-based, cluster randomised, placebo-controlled trial. AB - BACKGROUND: Zinc supplementation can reduce subsequent morbidity in children recovering from diarrhoea and respiratory illness in developing countries. However, whether routine supplementation would decrease morbidity and mortality in populations with zinc deficiency is unclear. We assessed the effect of daily zinc supplementation on children in southern Nepal. METHODS: We did a community based, cluster-randomised, double-masked, placebo-controlled, 2x2 factorial trial in children aged 1-35 months. Treatment groups were placebo, iron and folic acid, zinc, and iron and folic acid with zinc, with daily doses of 12.5 mg iron, 50 microg folic acid, and 10 mg zinc. Study staff gave children tablets on 2 days each week and left tablets with caregivers for other days. All children received vitamin A supplementation twice per year. Results of the iron arm of the trial have been reported previously. Between October, 2001, and January, 2006, 41,276 children were enrolled into the placebo (n=20,308) or zinc (n=20,968) groups and were followed-up for 60,636.3 person-years. The primary outcome was child mortality, and analyses were by intention to treat. Daily reports of signs and symptoms of common morbidities in stratified random subsamples of children were assessed every week for 12 months. This study is registered at ClinicalTrials.gov, number NCT00109551. FINDINGS: 2505 children refused to continue the trial and 3219 children were lost to follow-up. There was no significant difference in mortality between the zinc and placebo groups (316 vs 333 deaths; hazard ratio 0.92, 95% CI 0.75-1.12). Zinc had no effect on mortality in children younger than 12 months (181 vs 168 deaths; 1.04, 0.83-1.31); mortality was lower, but not statistically significantly so, in older children receiving zinc (135 vs 165; 0.80, 0.60-1.06). The frequency and duration of diarrhoea, persistent diarrhoea, dysentery, and acute lower respiratory infections did not differ between the groups. INTERPRETATION: Total mortality of children receiving zinc supplementation was not significantly different from that of children receiving placebo. Further data are needed from other populations with endemic zinc deficiency to confirm the potential age-specific effects reported in this study. PMID- 17920920 TI - Seizure and prominent abdominal veins. PMID- 17920921 TI - Comparison of Charlson comorbidity index and Kaplan-Feinstein index in patients with stage I lung cancer after surgical resection. AB - OBJECTIVE: We sought to determine whether Charlson comorbidity index (CCI) or Kaplan-Feinstein index (KFI) is a better predictor of prognosis in patients with stage I NSCLC after surgical resection. METHODS: A retrospective study of medical records of 426 patients with stage I lung cancer having complete surgical resection from 1995 to 2000 was performed. Data collected included age, gender, smoking history, resection type, pleural invasion status, and tumor type and size. Comorbidity score was determined using Charlson comorbidity index and Kaplan-Feinstein index. Both univariate and multivariate analyses were used to evaluate prognostic factors. RESULTS: Three hundred and twenty-eight male (76.99%) and 98 female (23.01%) patients had a mean age of 67.07 years (range 19 88 years). Median duration of follow-up was 60.32 months. Total follow-up rate was 95.1%. Distribution of CCI score was: 0, 236 (55.40%); 1, 112 (26.29%); >or=2, 78 (18.31%). Overall KFI score was: none, 247 (57.98%); mild, 126 (29.58%); moderate, 43 (10.09%); and severe, 10 (2.35%). In univariate analyses, patients aged>or=65 years, male, smokers, CCI score>or=2, extensive resection and pathological stage IB cancer had poorer 5-year survival. In multivariate logistic regression analysis, age>or=65 years, pneumonectomy, CCI score>or=2, and stage IB cancer were independent prognostic factors for poorer 5-year survival. CONCLUSIONS: Patients with CCI>or=2 had higher perioperative mortality and death from non-cancer causes after surgery compared to patients with CCI<2. However, KFI score had no impact on operative mortality and non-cancer death during follow up. PMID- 17920922 TI - Translocation of dilated pulmonary artery for relief of bronchial compression associated with ventricular septal defect. AB - Bronchial compression due to a dilated pulmonary artery is a troublesome problem in the surgical management of infants or children with congenital heart disease. We experienced an infantile case of ventricular septal defect and prolonged respiratory insufficiency caused by right bronchial compression and left pulmonary hypoplasia. Anterior translocation of the dilated right pulmonary artery and intracardiac repair succeeded in relieving the bronchial compression and improving left pulmonary function. We advocate that this procedure is useful for bronchial compression with congenital heart disease and maldevelopment of the lung. PMID- 17920924 TI - The surgical treatment of end-stage heart failure. AB - Cardiac failure remains the leading cause of death in the Western World today. After myocardial insult, as the heart remodels and dilates, an increase in wall tension occurs secondary to increased radius of curvature, leading to increased myocardial oxygen consumption, decreased subendocardial blood flow, impaired energetics, and increased arrhythmias. Poor prognosis directly correlates with the degree of remodeling. Despite improvements in left ventricular function and long-term outcomes seen with pharmacologic therapy, the results remain far from perfect and the mortality continues to be high. The surgical armamentarium for treating end-stage heart failure is broad and new types of surgical treatments continue to emerge as alternatives to cardiac transplantation for the treatment of end-stage heart failure. Furthermore, surgical therapies that were once contraindicated for use in the failing heart are now being used to halt or reverse ventricular remodeling and improve cardiac function. Therefore, an aggressive approach to surgical revascularization, correction of mitral insufficiency, surgical reversal of left ventricular remodeling, and long-term use of mechanical ventricular assistance should be considered in any heart failure patient who has exhausted pharmacologic therapy. PMID- 17920925 TI - Anti-inflammatory effects of low-level laser therapy (LLLT) with two different red wavelengths (660 nm and 684 nm) in carrageenan-induced rat paw edema. AB - It has been suggested that low-level laser therapy (LLLT) can modulate inflammatory processes. The aim of this experiment was to investigate what effects red laser irradiation with two different wavelengths (660 nm and 684 nm) on carrageenan-induced rat paw edema and histology. Thirty two male Wistar rats were randomly divided into four groups. One group received a sterile saline injection, while inflammation was induced by a sub-plantar injection of carrageenan (1 mg/paw) in the three other groups. After 1 h, LLLT was administered to the paw in two of the carrageenan-injected groups. Continuous wave 660 nm and 684 nm red lasers respectively with mean optical outputs of 30 mW and doses of 7.5 J/cm(2) were used. The 660 nm and 684 nm laser groups developed significantly (p<0.01) less edema (0.58 ml [SE+/-0.17] ml and 0.76 ml [SE+/-0.10] respectively) than the control group (1.67 ml [SE+/-0.19]) at 4h after injections. Similarly, both laser groups showed a significantly lower number of inflammatory cells in the muscular and conjunctive sub-plantar tissues than the control group. We conclude that both 660 nm and 684 nm red wavelengths of LLLT are effective in reducing edema formation and inflammatory cell migration when a dose of 7.5 J/cm(2) is used. PMID- 17920926 TI - Head-to-head comparison of BNP and IL-6 as markers of clinical and experimental heart failure: Superiority of BNP. AB - Activation of BNP and IL-6 are hallmarks of left ventricular (LV) dysfunction and congestive heart failure (CHF). To assess the relative activation of BNP and IL-6 in clinical and experimental heart failure, we performed a human study in which plasma N-terminal proBNP (NT-proBNP) and IL-6 were measured in a large group of patients in the chronic phase after myocardial infarction (MI) and an animal study in which LV gene expression of BNP and IL-6 was assessed in rapid ventricular pacing-induced heart failure. In the human study, NT-proBNP and IL-6 were measured by non-extracted, enzyme-linked immunoassay in 845 subjects (n=468 outpatients after MI, MONICA MI register Augsburg; and 377 siblings without MI, control). NT-proBNP (295+/-23pg/mL vs. CTRL 84+/-8, P<0.05) and IL-6 (2.7+/ 0.1pg/mL vs. CTRL 2.1+/-0.1, P<0.05) were both elevated in subjects with MI. These increases were particularly pronounced in the presence of concomitant CHF (both P<0.01 vs. CTRL) and LV dysfunction (EF<45%, both P<0.05 vs. CTRL). However, NT-proBNP was significantly correlated with several cardiac structural and functional parameters (EF, LVMI, history of MI, CHF symptoms; all P<0.05) upon regression analysis whereas IL-6 was only correlated with history of MI (P<0.001). Accordingly, MI subjects with symptomatic LV dysfunction were detected by NT-proBNP with a greater sensitivity, specificity, and ROC-area (85%, 88%, and 0.87, respectively) as compared to IL-6 (69%, 53%, and 0.67, respectively). In the animal study, IL-6 and BNP expression were both significantly elevated in CHF (both P<0.05) but with a much greater absolute activation of BNP. In addition, BNP mRNA expression displayed a stronger inverse correlation with LV function (r= 0.74; P<0.001) than IL-6 (r=-0.53; P=0.001) and was a markedly more sensitive and specific molecular marker of LV dysfunction (sensitivity 91%, specificity 100%, ROC-area 0.94) than IL-6 (sensitivity 74%, specificity 83%, ROC-area 0.87). Our animal study provides evidence that IL-6 expression is activated in heart failure but to a significantly lesser degree than that of BNP. Both the stronger expression of BNP and the better correlation with LV function provide the molecular basis for a diagnostic superiority of NT-proBNP in clinical LV dysfunction and heart failure. PMID- 17920927 TI - Genotoxicity and carcinogenicity studies of benzodiazepines. AB - This survey is a compendium of genotoxicity and carcinogenicity information of benzodiazepines and benzodiazepine analogues. Data from 51 drugs were collected; 41 of them are still in the market. Of the 51 drugs, 12 (23.5%) do not have retrievable genotoxicity or carcinogenicity data. The remaining 39 (76.5%) have at least one genotoxicity or carcinogenicity test result. Of these 39, 12 (30.8%) have at least one positive finding: 9 tested positive in at least one genotoxicity assay, 8 in at least one carcinogenicity assay, and 5 gave a positive result in both at least one genotoxicity assay and at least one carcinogenicity assay. In terms of correlation between results of the various genotoxicity assays and absence of carcinogenic activity in mice and/or rats or in other species, none of the 11 non-carcinogenic drugs tested positive in one of the various genotoxicity assay systems. Concerning the predictivity of genetic toxicology findings for the result(s) of long-term carcinogenesis assays, 18 drugs had both genotoxicity and carcinogenicity data; of these 11 (61.1%) were neither genotoxic nor carcinogenic, 2 (11.1%) were carcinogenic in at least one sex of mice or rats but tested negative in genotoxicity assays, and 5 (27.8%) gave a positive response in at least one genotoxicity assay and in at least one carcinogenicity assay. Only 8 (19.5%) of the 41 marketed benzodiazepines and benzodiazepine analogues had all data required by current guidelines for testing of pharmaceuticals. PMID- 17920928 TI - Bimolecular and unimolecular contributions to the disparate self-chemical ionizations of alpha-pinene and camphene isomers. AB - The contributions of molecular and fragment ions toward the disparate self chemical ionization (SCI) of alpha-pinene and camphene isomers were investigated. A kinetic model was constructed to predict the SCI outcomes for these two C(10)H(16) isomers. A major portion of the camphene molecular ions (isolated 500 ms after the 10 ms EI event at 24 eV) unimolecularly dissociated within 200 s of the ionization event. Conversely, under similar experimental conditions, the alpha-pinene molecular ions as well as the major fragment ions of alpha-pinene and camphene showed no unimolecular dissociation. The alpha-pinene and camphene molecular ions yielded product ions through two different reaction mechanisms (direct charge-transfer {CT} and indirect proton transfer {PT}). The isolated terpene fragment ions at m/z 93 and 121 reacted with their respective neutrals to produce [M + H](+). Proton affinity (PA) bracketing experiments, PA additivity schemes, and alkene PA versus adiabatic ionization energy (IE) linear correlation indicated that the PAs of camphene and alpha-pinene were comparable ( approximately 210 +/- 2 kcal x mol(-1)). The observed [M + H](+) SCI terpene ions were mainly the products of various fragment ion reactions. PMID- 17920929 TI - Stability engineering, biophysical, and biological characterization of the myeloid activating receptor immunoglobulin-like transcript 1 (ILT1/LIR-7/LILRA2). AB - Immunoglobulin-like transcript 1 (ILT1/LIR-7/LILRA2/CD85h) is one of the activating receptors in the ILT family whose members have been reported to regulate a broad range of cells involved in the immune response. Although inhibitory ILT receptors have been extensively studied, however, functions and structures of ILT activating receptors have yet to be elucidated. Obtaining of sufficient amount of recombinant proteins is a requisite for the functional and structural studies of a given protein. As a technical bottleneck of the study, extracellular domains of the ILT1 form aggregation during recombinant production in the past efforts. Here, we report the large-scale stable production of ILT1 D1D2 domains through engineering of site-directed mutagenesis (R142C) that introduces a cysteine at amino acid position 142 to form a disulfide bond with the spare cys132 without topological influences of the native protein based on the known structures of the homologous ILT 2/4/11. The recombinant ILT1 D1D2 domains behave as an equilibrium of both stable dimer and monomer in solution and yield ideal crystals for structural determination. The availability of quantities of soluble ILT1 D1D2 domains provides useful reagent for further studies of its detailed structure and functions. PMID- 17920930 TI - Phenoloxidase activity in the hemolymph of the spiny lobster Panulirus argus. AB - The prophenoloxidase activating system plays a major role in the defense mechanism of arthropods. In the present study, the phenoloxidase activity and its location in the hemolymph of the spiny lobster Panulirus argus is presented. Phenoloxidase activity was observed in the hemocyte lysate supernatant (HLS) and plasma after their incubation with trypsin. Higher amounts of trypsin were required to activate the HLS prophenoloxidase, due to the presence of a trypsin inhibitor in this fraction. Activation of prophenoloxidase was found when HLS was incubated with calcium, with an optimal pH between 7.5 and 8. This spontaneous activity is due to the prophenoloxidase activating enzyme, a serine proteinase that activates the prophenoloxidase once calcium ions were available. SDS was able to induce phenoloxidase activity in plasma and hemocyte fractions. Prophenoloxidase from HLS occurs as an aggregate of 300kDa. Electrophoretic studies combining SDS-PAGE and native PAGE indicate that different proteins produced the phenoloxidase activity found in HLS and plasma. Thus, as in most crustaceans, Panulirus argus contains a prophenoloxidase activating system in its hemocyte, comprising at least the prophenoloxidase activating enzyme and the prophenoloxidase. Finally, it is suggested that phenoloxidase activity found in plasma is produced by hemocyanin. PMID- 17920931 TI - Simulation-based evaluation of OSEM iterative reconstruction methods in dynamic brain PET studies. AB - The reconstruction of dynamic PET data is usually performed using filtered backprojection algorithms (FBP). This method is fast, robust, linear and yields reliable quantitative results. However, the use of FBP for low count data, such as dynamic PET data, generally results in poor visual image quality, exhibiting high noise, disturbing streak artifacts and low contrast. These signal-to-noise ratio and contrast in the reconstructed images may alter the quantification of physiological indexes, such as the regional Binding Potential (BP) obtained from kinetic modeling. Iterative reconstruction methods are often presented as viable alternatives to FBP reconstruction. In this study, we investigated the characteristics of the UW-OSEM and the ANW-OSEM iterative reconstruction methods in the context of ligand-receptor PET studies with low counts. The assessment was conducted using replicates of simulated [18F]MPPF acquisitions. The quantitative accuracy obtained with the iterative and analytical methods was compared. The results show that analytical methods are more robust to the low count data than iterative methods, and therefore enable a better estimate of the regional activity values and binding potential. The positivity constraint in MLEM-based algorithms leads to overestimations of the activity in regions with low activity concentration, typically the cerebellum. This overestimation results in significant bias in BP estimates with iterative reconstruction methods. The bias is confirmed from the reconstruction of real PET data. PMID- 17920932 TI - Motivation and semantic context affect brain error-monitoring activity: an event related brain potentials study. AB - During speech production, we continuously monitor what we say. In situations in which speech errors potentially have more severe consequences, e.g. during a public presentation, our verbal self-monitoring system may pay special attention to prevent errors than in situations in which speech errors are more acceptable, such as a casual conversation. In an event-related potential study, we investigated whether or not motivation affected participants' performance using a picture naming task in a semantic blocking paradigm. Semantic context of to-be named pictures was manipulated; blocks were semantically related (e.g., cat, dog, horse, etc.) or semantically unrelated (e.g., cat, table, flute, etc.). Motivation was manipulated independently by monetary reward. The motivation manipulation did not affect error rate during picture naming. However, the high motivation condition yielded increased amplitude and latency values of the error related negativity (ERN) compared to the low-motivation condition, presumably indicating higher monitoring activity. Furthermore, participants showed semantic interference effects in reaction times and error rates. The ERN amplitude was also larger during semantically related than unrelated blocks, presumably indicating that semantic relatedness induces more conflict between possible verbal responses. PMID- 17920933 TI - Spatiotemporal dynamics of audiovisual speech processing. AB - The cortical processing of auditory-alone, visual-alone, and audiovisual speech information is temporally and spatially distributed, and functional magnetic resonance imaging (fMRI) cannot adequately resolve its temporal dynamics. In order to investigate a hypothesized spatiotemporal organization for audiovisual speech processing circuits, event-related potentials (ERPs) were recorded using electroencephalography (EEG). Stimuli were congruent audiovisual/ba/, incongruent auditory/ba/synchronized with visual/ga/, auditory-only/ba/, and visual only/ba/and/ga/. Current density reconstructions (CDRs) of the ERP data were computed across the latency interval of 50-250 ms. The CDRs demonstrated complex spatiotemporal activation patterns that differed across stimulus conditions. The hypothesized circuit that was investigated here comprised initial integration of audiovisual speech by the middle superior temporal sulcus (STS), followed by recruitment of the intraparietal sulcus (IPS), followed by activation of Broca's area [Miller, L.M., d'Esposito, M., 2005. Perceptual fusion and stimulus coincidence in the cross-modal integration of speech. Journal of Neuroscience 25, 5884-5893]. The importance of spatiotemporally sensitive measures in evaluating processing pathways was demonstrated. Results showed, strikingly, early (<100 ms) and simultaneous activations in areas of the supramarginal and angular gyrus (SMG/AG), the IPS, the inferior frontal gyrus, and the dorsolateral prefrontal cortex. Also, emergent left hemisphere SMG/AG activation, not predicted based on the unisensory stimulus conditions was observed at approximately 160 to 220 ms. The STS was neither the earliest nor most prominent activation site, although it is frequently considered the sine qua non of audiovisual speech integration. As discussed here, the relatively late activity of the SMG/AG solely under audiovisual conditions is a possible candidate audiovisual speech integration response. PMID- 17920935 TI - Modelling vascular reactivity to investigate the basis of the relationship between cerebral blood volume and flow under CO2 manipulation. AB - Changes in cerebral blood flow (f) and vascular volume (v) are of major interest in mapping cerebral activity and metabolism, but the relation between them currently lacks a sufficient theoretical basis. To address this we considered three models: a uniform reactive tube model (M1); an extension of M1 that includes passive arterial inflow and venous volume (M2); and a more anatomically plausible model (M3) consisting of 19 compartments representing the whole range of vascular sizes and respective CO2 reactivities, derived from literature data. We find that M2 cannot be described as the simple scaling of a tube law, but any divergence from a linear approximation is negligible within the narrow physiological range encountered experimentally. In order to represent correctly the empirically observed slope of the overall v-f relationship, the reactive bed should constitute about half of the total vascular volume, thus including a significant fraction of capillaries and/or veins. Model M3 demonstrates systematic variation of the slope of the v-f relationship between 0.16 and 1.0, depending on the vascular compartment under consideration. This is further complicated when other experimental approaches such as flow velocity are used as substitute measurements. The effect is particularly large in microvascular compartments, but when averaged with larger vessels the variations in slope are contained within 0.25 to 0.55 under conditions typical for imaging methods. We conclude that the v-f relationship is not a fixed function but that both the shape and slope depend on the composition of the reactive volume and the experimental methods used. PMID- 17920936 TI - Cryptic speciation and paraphyly in the cosmopolitan bryozoan Electra pilosa- impact of the Tethys closing on species evolution. AB - Cosmopolitan nature of the marine bryozoan Electra pilosa was studied to clarify geographic structure and to outline evolution and phylogeography of the species. Several local populations from the Northeast Atlantic (North Sea and Baltic Sea), Arctic (Barents Sea and White Sea) and Indo-West Pacific (New Zealand) were compared. In addition, we examined the closely related species E. posidoniae from the Mediterranean Sea. Phylogenetic analysis based on both 16S and 18S rDNA indicate that the Indo-West Pacific E. pilosa is a sister species to the Atlantic Mediterranean clade, with the latter including the species E. posidoniae and the Atlantic population of E. pilosa. The topology of the phylogenetic tree leads us to conclude that E. pilosa is a paraphyletic species group relative to E. posidoniae, and a molecular dating of its divergence is consistent to geologic events associated with the closure of the Tethys Sea. PMID- 17920937 TI - Molecular phylogenetic analysis of class Colpodea (phylum Ciliophora) using broad taxon sampling. AB - The ciliate class Colpodea provides a powerful case in which a molecular genealogy can be compared to a detailed morphological taxonomy of a microbial group. Previous analyses of the class using the small-subunit rDNA are based on sparse taxon sampling, and are therefore of limited use in comparisons with morphologically-based classifications. Taxon sampling is increased here to include all orders within the class, and more species within previously sampled orders and in the species rich genus Colpoda. Results indicate that the Colpodea may be paraphyletic, although there is no support for deep nodes. The orders Bursariomorphida, Grossglockneriida, and Sorogenida are monophyletic. The orders Bryometopida, Colpodida, and Cyrtolophosidida, and the genus Colpoda, are not monophyletic. Although congruent in many aspects, the conflict between some nodes on this single gene genealogy and morphology-based taxonomy suggests the need for additional markers as well as a reassessment of the Colpodea taxonomy. PMID- 17920934 TI - Persistent neural activity during the maintenance of spatial position in working memory. AB - The mechanism for the short-term maintenance of information involves persistent neural activity during the retention interval, which forms a bridge between the cued memoranda and its later contingent response. Here, we used event-related functional magnetic resonance imaging to identify cortical areas with activity that persists throughout working memory delays with the goal of testing if such activity represents visuospatial attention or prospective saccade goals. We did so by comparing two spatial working memory tasks. During a memory-guided saccade (MGS) task, a location was maintained during a delay after which a saccade was generated to the remembered location. During a spatial item recognition (SIR) task identical to MGS until after the delay, a button press indicated whether a newly cued location matched the remembered location. Activity in frontal and parietal areas persisted above baseline and was greater in the hemisphere contralateral to the cued visual field. However, delay-period activity did not differ between the tasks. Notably, in the putative frontal eye field (FEF), delay period activity did not differ despite that the precise metrics of the memory guided saccade were known during the MGS delay and saccades were never made in SIR. Persistent FEF activity may therefore represent a prioritized attentional map of space, rather than the metrics for saccades. PMID- 17920938 TI - Validation of respiratory safety pharmacology models: conscious and anesthetized beagle dogs. AB - INTRODUCTION: Installation, operation and performance qualifications were performed on a test system for respiratory monitoring. METHODS: For performance qualification, conscious dogs received saline (0.2 mL/kg, iv, n=12), albuterol (100 microg/kg, inhalation, n=5), methacholine (2.0 and 8.0 microg/kg, iv, n=8) and remifentanil (4.0 microg/kg, iv, n=7). Following anesthesia with propofol infusion, dogs received saline (iv, n=15), albuterol (100 microg/kg, inhalation, n=8), methacholine (8.0 microg/kg, iv, n=8), remifentanil (4.0 microg/kg, iv, n=7), and cholecystokinine tetrapeptide (CCK-4) (10 microg/kg, iv, n=7) and were exposed to hypoxic gas mixture (10% oxygen) (n=12). RESULTS: Saline had no significant respiratory effect. Albuterol increased tidal volume (TV) (+28%, p<0.05) and minute ventilation (MV) (+96%, p<0.01) in conscious dogs. In anesthetized dogs, MV was significantly increased (+23%, p<0.05) but the difference was not statistically significant for TV and respiratory rate (RR). Methacholine at 2.0 microg/kg increased MV (+45%, p<0.01) in conscious animals while 8.0 microg/kg increased RR (+66%, p<0.01), TV (+24%, p<0.05) and MV (+88%, p<0.05). In anesthetized dogs, methacholine increased RR (+51%, p<0.05), MV (+34%, p<0.05), lung elastance (+36.9%, p<0.01), and resistance (+45.8%, p<0.01). Remifentanil decreased MV in conscious dogs (-68%, p<0.01) while transient apnea was observed in all anesthetized dogs. CCK-4 increased RR (+328%, p<0.01) and MV (+127%, p<0.05) and decreased TV (-58%, p<0.01). Exposure to hypoxic gas mixture increased MV and RR (p<0.01). Baseline MV was lower (p<0.05) in anesthetized than in conscious dogs. DISCUSSION: Arterial blood gas values, particularly SaO(2), presented a limited sensitivity to detect any ventilation disturbance, but allowed confirmation of both ventilatory compensatory phenomenon (when present) and initial pharmacologic drug effect. These results also highlight the greater sensitivity of the conscious model when compared to anesthetized dogs. PMID- 17920939 TI - In vitro expansion affects the response of chondrocytes to mechanical stimulation. AB - OBJECTIVE: Expansion of autologous chondrocytes is a common step in procedures for cartilage defect repair. Subsequent dedifferentiation can alter cellular response to mechanical loading, having major consequences for the cell's behavior in vivo after reimplantation. Therefore, we examined the response of primary and expanded human articular chondrocytes to mechanical loading. METHOD: Primary and expanded chondrocytes were stretched at either 0.5% or 3.0% at 0.5Hz, 2h per day, for 3 days. Gene expression levels of matrix components (aggrecan (AGC1), lubricin (PRG4), collagen type I (COL1), type II (COL2) and type X (COL10)) as well as matrix enzymes (matrix metalloproteinase 1 (MMP1), MMP3, MMP13) and SOX9 were compared to unstretched controls. To evaluate the effect of a chondrogenic environment on cellular response to stretch, redifferentiation medium was used on expanded cells. RESULTS: In primary chondrocytes, stretch led to mild decreases in AGC1, COL1 and COL10 gene expression (maximum of 3.8-fold) and an up regulation of PRG4 (2.0-fold). In expanded chondrocytes, expression was down regulated for AGC1 (up to 21-fold), PRG4 (up to 5.0-fold), COL1 (10-fold) and COL2 (2.9-fold). Also, expression was up-regulated for MMP1 (20-fold) and MMP3 (up to 4-fold), while MMP13 was down-regulated (2.8-fold). A chondrogenic environment appeared to temper effects of stretch. DISCUSSION: Our results show that expansion alters the response of human chondrocytes to stretch. Expanded chondrocytes greatly decrease gene expression of matrix constituents and increase expression of MMPs, whereas primary chondrocytes hardly respond. Our data could be a reference for optimization of cell sources or expansion protocols for reimplanted chondrocytes. PMID- 17920940 TI - The validity of in vivo ultrasonographic grading of osteoarthritic femoral condylar cartilage: a comparison with in vitro ultrasonographic and histologic gradings. AB - OBJECTIVES: To establish an ultrasonographic (US) grading for semiquantitative evaluation of the femoral condylar cartilage of knee osteoarthritis (OA), in vivo, and compare the in vivo US grading with the in vitro US and histologic gradings. DESIGN: Ninety-five patients going to receive total knee arthroplasty because of OA of the knee were recruited. US examination was performed in vivo in the day before operation using a grading system including parameters of margin sharpness, clarity and thickness. Specimens of the medial and lateral distal femoral condyles taken during the operation were graded with in vitro US and histologic evaluation. The correlation between the in vivo US and in vitro US as well as between the in vivo US and histologic gradings was analyzed. RESULTS: In 172 femoral condyles (including medial and lateral ones), the distribution of grading ranged from Grade 1 to 6 in in vivo US and from Grade 1 to 4 in histologic examination. The in vivo US grading was significantly correlated to in vitro US grading over anterior and middle areas (p<0.001, Rho=0.35 and 0.45, respectively) and histologic grading over these two areas (p<0.001, Rho=0.40 and 0.36, respectively). When the cases with maximal angle of knee flexion less than 120 degree were excluded, the correlation was better. CONCLUSIONS: The significant correlation between in vivo US and histologic gradings might permit semi-quantitative in vivo US assessment of osteoarthritic femoral condylar cartilage. PMID- 17920941 TI - Immunological aspects of human amniotic fluid cells: implication for normal pregnancy. AB - Amniotic fluid cells (AFCs) are routinely obtained and expanded in vitro for prenatal diagnosis; nevertheless current knowledge about their properties is limited. The detailed mechanisms underlying normal pregnancies are yet to be discovered. The goal of this study was to identify the immunological aspects of AFCs including cytokine production and human leukocyte antigen (HLA) expression, and to discuss its implication for pregnancy. Eighty-six samples of AFCs were determined for HLA expression before and after culture. Cytokine production was measured with flow cytometry in AFC culture supernatants. Treatment of interferon (IFN)-gamma on induction of HLA-DR expression in cultured AFCs was also investigated. Data indicated that both fresh and cultured AFCs express HLA-I, HLA G, but not HLA-DR, and the cultured AFCs predominately produce the cytokine interleukin (IL)-6. Importantly, we observed that IFN-gamma could induce HLA-DR expression on cultured AFCs in a dose-dependent manner. Taken together, our results indicated that AFCs are functionally active cells and are significant in pregnancy. PMID- 17920942 TI - Effect of nicotine and polyaromtic hydrocarbons on cerebral endothelial cells. AB - The present study was designed to investigate the effect of nicotine and polyaromatic hydrocarbon compounds on cerebral endothelial cells (CECs). Nicotine treatments from 15 min to 5h did not cause any changes in the expression and localization of principal junctional proteins. One day of treatment with a relatively high concentration of nicotine induced a decrease in the expression of the tight junction protein ZO-1, occludin, and the adherens junction protein, cadherin. Treatment with 3 x 10(-5)M phenanthrene for 24h caused a redistribution of occludin from the Triton X-100 insoluble to the Triton X-100 soluble fraction. Transendothelial electrical resistance was not significantly affected by 24h treatments with nicotine, methylanthracene or phenanthrene. However, 24h nicotine treatment increased transendothelial permeability in CECs exposed to oxidative stress. Both nicotine and phenanthrene were able to regulate the expression of a large number of proteins as revealed by 2D electrophoresis. Our experiments suggest that tobacco smoking may affect the junctional complex of CECs, and that this effect is enhanced by oxidative stress. PMID- 17920943 TI - Dihydrorhodamine 123 is superior to 2,7-dichlorodihydrofluorescein diacetate and dihydrorhodamine 6G in detecting intracellular hydrogen peroxide in tumor cells. AB - Dihydrorhodamine 123 (DHR 123), 2,7-dichlorodihydrofluorescein diacetate (DCFH DA), and dihydrorhodamine 6G (DHR 6G) were evaluated as probes for detecting cellular hydrogen peroxide levels in SPC-A-1 lung adenocarcinoma cells. Imaging techniques and fluorescence-activated cell scan were used in the study of the probe responses. Obvious green fluorescence was established after a 25-min exposure. After staining with MitoTracker Orange CM-H2TMRos (a probe for mitochondria) and the abovementioned probes simultaneously, only the DHR 123 and DHR 6G groups exhibited legible green fluorescence in the mitochondrial regions. Furthermore, the DHR 6G group exhibited weaker fluorescence intensity. When 100 microM H2O2 was added to SPC-A-1 cells loaded with these probes, the intracellular fluorescence increased rapidly and significantly. Our results suggest that DHR 123 is superior for the instantaneous detection of cellular hydrogen peroxide in SPC-A-1 cells. PMID- 17920944 TI - Bak Foong protects dopaminergic neurons against MPTP-induced neurotoxicity by its anti-apoptotic activity. AB - Bak Foong pill (BFP) is a well-known traditional Chinese medicine used for treatment of various gynaecological disorders. In addition, it exerts beneficial effects on other functional systems including the central nervous system. In the present study, we have investigated the possible neuroprotective action of BFP upon the nigrostriatal dopaminergic system by examining its effect on the expression patterns of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the 1-methyl-4-phenyl-1,2,3,6-tetrahyrdropyridine (MPTP)-induced Parkinson's disease (PD) mouse model. MPTP significantly decreased TH and DAT mRNA levels in the striatum and midbrain of both female and male C57BL/6 mice. However, with BFP pre-treatment mice showed a reduced neurotoxicity, with TH and DAT mRNA levels either not affected by MPTP or affected to a lesser extent in the midbrain and striatum when compared to vehicle treated animals. Possible anti-apoptotic activity of BFP was further studied in a dopamine-secreting neuroendocrine cell line, PC12. In this assay, MPTP elevated the expression of a pro-apoptotic gene, Bax, while this expression was reduced by BFP pre-treatment. Flow cytometry results also revealed that the effect of MPTP-induced apoptosis in PC12 cell lines was significantly reduced by BFP. The present results suggest that BFP is able to protect dopaminergic neurons from neurotoxin-induced neuronal injury with anti-apoptotic activity being one of the possible mechanisms. PMID- 17920945 TI - Subcellular distribution of spermidine/spermine N1-acetyltransferase. AB - The subcellular distribution of the polyamine catabolic enzyme spermidine/spermine N(1)-acetyltransferase (SSAT) was studied in L56Br-C1 cells treated with 10 microM N(1),N(11)-diethylnorspermine (DENSPM) for 24 h. Cells were fractioned into three subcellular fractions. A particulate fraction containing the mitochondria was denoted as the mitochondrial fraction. After DENSPM treatment, an increase in SSAT activity was mainly found in the mitochondrial fraction. Western blot analysis showed an increased level of the SSAT protein in the mitochondrial fraction compared to the cytosolic fraction. Immunofluorescence microscopy and immunogold labeling transmission electron microscopy also showed a mitochondrial association of SSAT. Transmission electron microscopy revealed that the endoplasmic reticulum was devoid of ribosomes in DENSPM-treated cells, in contrast to control cells that contained ample ribosomes. An increased SSAT activity in connection with the mitochondria may be part of the mechanism of DENSPM-induced apoptosis. PMID- 17920946 TI - Novel anti-apoptotic effect of Bcl-2: prevention of polyamine depletion-induced cell death. AB - The spermine analogue N(1),N(11)-diethylnorspermine (DENSPM) efficiently depletes the polyamine pools in the breast cancer cell line L56Br-C1 and induces apoptotic cell death via the mitochondrial pathway. In this study, we have over-expressed the anti-apoptotic protein Bcl-2 in L56Br-C1 cells and investigated the effect of DENSPM treatment. DENSPM-induced cell death was significantly reduced in Bcl-2 over-expressing cells. Bcl-2 over-expression reduced DENSPM-induced release of the pro-apoptotic proteins AIF, cytochrome c, and Smac/DIABLO from the mitochondria. Bcl-2 over-expression reduced the DENSPM-induced activation of caspase-3. Bcl-2 over-expression also prevented DENSPM-induced Bax cleavage and reduction of Bcl-X(L) and survivin levels. The DENSPM-induced activation of the polyamine catabolic enzyme spermidine/spermine N(1)-acetyltransferase was reduced by Bcl-2 over-expression, partly preventing polyamine depletion. Thus, Bcl-2 over expression prevented a number of DENSPM-induced apoptotic effects. PMID- 17920950 TI - Biology of autograft and allograft healing in anterior cruciate ligament reconstruction. AB - The graft-bone attachment site is the weak link in anterior cruciate ligament (ACL) reconstruction surgery because healing is slow and often incomplete. Each of the many graft options for ACL reconstruction surgery has different healing characteristics and potential. Autografts that allow bone-to-bone healing offer the best healing potential. An understanding of the biology of graft healing should give the surgeon context in graft selection and rehabilitation for ACL reconstruction. PMID- 17920947 TI - Phenotypically and functionally distinct subsets of natural killer cells in human PBMCs. AB - Human natural killer (NK) cells are one major component of lymphocytes that mediate early protection against viruses and tumor cells, and play an important role in immune regulatory functions. In this study, we demonstrated that human NK cells could be divided into four subsets, CD56hi CD16(-), CD56lo CD16(-), CD56+CD16+ and CD56(-)CD16+, based on the expression of cell surface CD56 and CD16 molecules. Phenotypic analysis of NK cell subsets indicated that the expression of activation markers, adhesion molecules, memory cell markers, inhibitory and activating receptors, and intracellular proteins (granzyme B and perforin) were heterogeneous. Following interleukin (IL)-2 stimulation, interferon-gamma was preferentially produced by CD56+CD16(-) NK cells and this subset showed more proliferative capacity. The cytolytic activity of both CD56+CD16(-) and CD56+/-CD16+ subsets could be augmented in response to IL-2. The data provided a new definition for NK cell subsets demonstrating their phenotypic and functional diversity and possible stage of NK cell differentiation in peripheral blood. PMID- 17920951 TI - Bone-patella tendon-bone autograft anterior cruciate ligament reconstruction. AB - The anterior cruciate ligament (ACL) serves an important stabilizing and biomechanical function for the knee. Reconstruction of the ACL remains one of the most commonly performed procedures in the field of sports medicine. Reconstruction of the ACL with bone-patella tendon-bone (BPTB) autograft secured with interference screw fixation has been the historical reference standard and remains the benchmark against which other methods are gauged. This article reviews the reconstruction of the ACL with BPTB autograft including the surgical technique, rationale for BTPB use, and outcomes. PMID- 17920952 TI - Primary anterior cruciate ligament reconstruction using contralateral patellar tendon autograft. AB - Primary ACL reconstruction using a contralateral patellar tendon autograft is an effective means of achieving symmetrical range of motion and strength after surgery. When the graft is harvested from the ipsilateral knee, the rehabilitation for the ACL graft and for the graft-donor site are different and have opposing goals. Rehabilitation for the ACL graft involves obtaining full range of motion, reducing swelling, and providing the appropriate stress to achieve graft maturation. Rehabilitation for the graft-donor site involves performing high-repetition strengthening exercises to regain size and strength, best achieved when begun immediately after surgery. PMID- 17920953 TI - Principles for using hamstring tendons for anterior cruciate ligament reconstruction. AB - The use of autogenous hamstring tendon as a graft source for anterior cruciate ligament (ACL) reconstruction continues to gain in popularity. The low harvest morbidity and excellent biomechanical graft properties coupled with improved fixation of soft tissue grafts are all reasons for excellent clinical outcomes of ACL reconstruction using hamstring tendons. In addition, surgeon awareness of the complications associated with poor tunnel placement and more exacting tunnel placement techniques help prevent roof and posterior cruciate ligament impingement and contribute to the successful outcomes of hamstring ACL constructs. PMID- 17920954 TI - Quadriceps tendon--a reliable alternative for reconstruction of the anterior cruciate ligament. AB - Anterior cruciate ligament (ACL) reconstruction surgery with the central third quadriceps tendon can yield a stable, high-functioning knee with little associated morbidity. Both the quadriceps tendon-patellar bone graft and the free tendon graft are reported to produce good to excellent outcomes at more than 2 years of follow-up. The decreased donor-site morbidity and absence of anterior knee pain suggest that the quadriceps free tendon autograft offers a reliable, pain-free, low-morbidity autograft alternative in ACL reconstruction. Recent data suggest that this graft may be the least morbid of the currently used ACL autograft reconstruction alternatives. PMID- 17920955 TI - Allograft safety in anterior cruciate ligament reconstruction. AB - Allograft tissue seems to provide an excellent option for reconstruction of the ACL in the primary and revision setting. Although in general the risks of using allograft tissue in ACL reconstruction are low, the consequences of complications associated with disease or infection transmission or of recurrent instability secondary to graft failure are large. Surgeons should provide patients with the information available regarding allograft risks and should have thorough knowledge of the source and preparation of the grafts by their tissue bank before implantation for ACL reconstruction. PMID- 17920956 TI - Fresh-frozen allograft anterior cruciate ligament reconstruction. AB - Reconstruction of the anterior cruciate ligament provides consistently good to excellent results allowing return to work and sport. Allograft tissue is an alternative to autografts when appropriate donor tissue is not available or its use is not advisable for other reasons. The technique and results for allograft use are similar to those for autograft, making its use appropriate in a variety of clinical scenarios. This article reviews the indications for allograft ACL reconstruction, graft options, and technique for allograft use. PMID- 17920957 TI - Freeze-dried allografts for anterior cruciate ligament reconstruction. AB - Freeze-dried allografts represent a viable and functional alternative to fresh frozen allograft and autograft constructs. Compared with fresh-frozen allograft constructs, freeze-dried soft tissue allograft constructs have many advantages including limited immunogenicity, ease of graft storage, comparable mechanical properties of soft tissue constructs, and the potential for improved biologic incorporation. This article reviews the fundamental processing of freeze-dried allografts and summarizes the clinical and basic science studies supporting the safe and effective use of freeze-dried allograft constructs for anterior cruciate ligament reconstruction. It also discusses potential directions of future research on tissue-engineered anterior cruciate ligament constructs using freeze dried tendon constructs. PMID- 17920959 TI - Clinical outcomes of allograft versus autograft in anterior cruciate ligament reconstruction. AB - Anterior cruciate ligament (ACL) injuries are the most common complete ligamentous injury to the knee. The optimal graft should be able to reproduce the anatomy and biomechanics of the ACL, be incorporated rapidly with strong initial fixation, and cause low graft-site morbidity. This article reviews the literature comparing the clinical outcomes following allograft and autograft ACL reconstruction and examines current issues regarding graft choice. PMID- 17920958 TI - Soft tissue allograft and double-bundle reconstruction. AB - The anterior cruciate ligament (ACL) is composed of two functional bundles, the anteromedial and posterolateral. Multiple biomechanical and clinical studies have demonstrated that the posterolateral bundle plays a critical role in rotatory stability of the knee. Anatomic double-bundle reconstruction of the ACL best restores knee function and kinematics when the ACL is ruptured. For double-bundle ACL reconstruction, the use of allograft is safe, minimizes graft harvest morbidity, expedites recovery, and is associated with successful clinical results in short-term follow-up. PMID- 17920960 TI - Peripheral versus aperture fixation for anterior cruciate ligament reconstruction. AB - Because both the young and aging population are showing increasing interest in sports participation, the number of sports related injuries and in particular anterior cruciate ligament (ACL) injuries have been increasing. Because of these injuries much time and energy has been focused on ACL reconstruction in order to return these individuals to their optimal level of participation in their sport. This article explores and reviews the concepts of ACL fixation location and how this affects the ultimate outcome of this reconstructive procedure. PMID- 17920961 TI - Biomechanics of intratunnel anterior cruciate ligament graft fixation. AB - Interference screw fixation of bone-patellar tendon-bone grafts now is considered the standard against which all ACL graft-fixation techniques are compared, but mechanical fixation of the ACL graft in the bone tunnels is the weak link in the early postoperative period. This article discusses some of the limitations of in vitro biomechanical studies and reviews variables that influence the tensile properties of intratunnel fixation methods for bone-tendon-bone and soft tissue grafts. PMID- 17920962 TI - Immune regulatory networks in the post-transplant setting. PMID- 17920964 TI - Cytokine mRNA responses in bovine epithelia during foot-and-mouth disease virus infection. AB - Foot-and-mouth disease (FMD) remains the single most important constraint to international trade in live animals and animal products. The factors which regulate the pathogenesis and persistence of foot-and-mouth disease virus (FMDV) are poorly understood. mRNA levels of the inflammatory cytokines interleukin (IL) 1alpha, tumour necrosis factor (TNF)-alpha and the antiviral cytokines interferon (IFN)-alpha, beta and gamma in microdissected epithelium from cattle acutely infected with FMDV O UKG 34/2001 were quantified using laser microdissection in combination with a quantitative reverse transcription polymerase chain reaction assay. Cytokine mRNA responses in microdissected epithelia from the bovine tongue, coronary band and dorsal soft palate during the acute stage of FMDV infection were different. Significantly increased expression of cytokine mRNA was found in microdissected epithelia from the coronary band and tongue of FMDV infected cattle and the extent of cytokine mRNA induction correlated with viral RNA loads. The results suggest that epithelia from different sites of an infected animal may mount different defences following infection and this may contribute to differences in their relative capacities to clear the virus. PMID- 17920965 TI - Transvenous electrical cardioversion of atrial fibrillation in six horses using custom made cardioversion catheters. AB - Pharmacological conversion of atrial fibrillation (AF) to sinus rhythm in horses can be difficult. The objective of this study was to investigate the feasibility of transvenous electrical cardioversion with custom made catheters in eight horses, of which three had failed cardioversion using quinidine sulfate. Two cardioversion catheters and one pacing/sensing electrode were inserted via the right jugular vein and placed using ultrasound guidance into the left pulmonary artery, the right atrium and the right ventricle, respectively. Because immediate recurrence of AF was encountered in the second horse treated, pre-treatment with amiodarone was given to each of the remaining six horses. Induction of general anaesthesia was associated with dislocation of the cardioversion catheter in three horses, requiring a second catheterisation procedure. During general anaesthesia, biphasic R wave synchronised shocks of up to 360 J were delivered between both cardioversion electrodes. In six horses (75%), including two which had failed quinidine sulfate treatment, sinus rhythm was restored with a mean energy level of 295+/-62 J. No side effects were observed. Blood analysis 3 h after cardioversion revealed normal parameters, including cardiac troponin I values. Transvenous electrical cardioversion of atrial fibrillation with custom made cardioversion catheters can be considered as a treatment option for atrial fibrillation in horses, especially when conventional drugs fail. PMID- 17920963 TI - Maintaining transparency: a review of the developmental physiology and pathophysiology of two avascular tissues. AB - The lens and cornea are transparent and usually avascular. Controlling nutrient supply while maintaining transparency is a physiological challenge for both tissues. During sleep and with contact lens wear the endothelial layer of the cornea may become hypoxic, compromising its ability to maintain corneal transparency. The mechanism responsible for establishing the avascular nature of the corneal stroma is unknown. In several pathological conditions, the stroma can be invaded by abnormal, leaky vessels, leading to opacification. Several molecules that are likely to help maintain the avascular nature of the corneal stroma have been identified, although their relative contributions remain to be demonstrated. The mammalian lens is surrounded by capillaries early in life. After the fetal vasculature regresses, the lens resides in a hypoxic environment. Hypoxia is likely to be required to maintain lens transparency. The vitreous body may help to maintain the low oxygen level around the lens. The hypothesis is presented that many aspects of the aging of the lens, including increased hardening, loss of accommodation (presbyopia), and opacification of the lens nucleus, are caused by exposure to oxygen. Testing this hypothesis may lead to prevention for nuclear cataract and insight into the mechanisms of lens aging. Although they are both transparent, corneal pathology is associated with an insufficient supply of oxygen, while lens pathology may involve excessive exposure to oxygen. PMID- 17920966 TI - Tissue factor activated thromboelastography correlates to clinical signs of bleeding in dogs. AB - The ability of a laboratory assay to correlate to clinical phenotype is crucial for the accurate diagnosis and monitoring of haemostasis and is therefore challenging with currently used routine haemostasis assays. Thromboelastography (TEG) is increasingly used to evaluate haemostasis in humans and may well be of value in the workup of dogs suspected of having a haemostatic disorder. This study was undertaken to evaluate prospectively how tissue factor (TF) activated TEG correlated to clinical signs of bleeding in dogs, compared to a routine coagulation profile. A prospective case-control study was performed over a 2 year period from 2004-2006. Eligible dogs were those where the primary clinician requested a coagulation profile to evaluate haemostasis. The dogs were simultaneously evaluated with a TF-activated TEG assay. Twenty-seven dogs, characterised as hypo-coagulable based on the TEG parameter G (<3.2 Kdyn/cm(2)), were included in the study as cases. Size matched control groups of TEG normo- (G=3.2K-7.2 Kdyn/cm(2)) and hyper-coagulable (G>7.2 Kdyn/cm(2)) dogs were selected retrospectively from the eligible dogs. For all dogs, clinical signs of bleeding were noted at time of analysis. There were statistically significant differences between all TEG values of hypo- and normo- and hyper-coagulable dogs. Thromboelastography correctly identified dogs with clinical signs of bleeding with a positive predictive value (PPV) of 89% and a negative predictive value (NPV) of 98% based on G alone. In comparison, the coagulation profile had a PPV between 50-81% and a NPV between 92-93% for detection of bleeding, depending on the observer. In conclusion, a TF-activated TEG G value<3.2K dyn/cm(2) correctly identified dogs with clinical signs of bleeding with very high PPV and NPV, irrespective of observer. The findings strongly suggest that TF- activated TEG may be of value in the workup of dogs suspected of having a haemostatic disorder. PMID- 17920967 TI - Molecular evaluation of the incidence of Ehrlichia canis, Anaplasma platys and Babesia spp. in dogs from Ribeirao Preto, Brazil. AB - Canine monocytic ehrlichiosis caused by Ehrlichiacanis is endemic in many regions of Brazil. Since thrombocytopenia is a common finding in infected dogs, many clinicians tend to use it as an indication for antibiotic treatment. Polymerase chain reaction (PCR) and nested PCR were used to study the presence of E. canis, Anaplasma platys and Babesia spp. in thrombocytopenic and non-thrombocytopenic dogs from Ribeirao Preto, Brazil. Despite the high prevalence of E. canis infection among thrombocytopenic dogs, 46.7% of the thrombocytopenic dogs studied were either infected with Babesia spp. or A.platys or not infected with any of the three pathogens. There was a high incidence (25.4%) of E. canis infection in non-thrombocytopenic dogs. Although infection with E. canis should be considered in thrombocytopenic dogs, the final diagnosis needs to be confirmed by complementary tests such as blood smears and PCR to avoid the unnecessary use of antibiotics. PMID- 17920969 TI - Triage flowchart to rule out acute coronary syndrome. AB - AIM: The aim of the study was to establish a triage flowchart to rule out acute coronary syndrome (ACS) among patients with chest pain (CP) arriving on an Emergency Department (ED). PATIENTS AND METHOD: This prospective observational study included 1000 consecutive patients with CP arriving on an ED CP unit. Demographic and clinical characteristics along with vital signs were recorded as independent variables. After CP unit protocol completion and 1-month follow-up, patients were classified as (dependent variable) (1) true non-ACS (all noncoronary patients at the first visit that kept this condition when called 1 month later) or (2) true ACS (all the remaining patients). Relationship among variables was assessed by multiple logistic regression analysis. A triage flowchart was obtained from significant variables and applied to patients with CP who were then grouped in "triage non-ACS" and "triage ACS." Validity indexes to exclude ACS for triage flowchart were measured. RESULTS: Variables significantly associated with non-ACS and included in the triage flowchart were age <40 years (odds ratio 3.61, 95% CI 1.63-7.99), absence of diabetes (2.74, 1.53-4.88), no previously known coronary artery disease (5.46, 3.42-8.71), nonoppressive pain (10.63, 6.04-18.70), and nonretrosternal pain (5.16, 2.82-9.42). For the triage flowchart, both specificity and positive predictive value to rule out ACS were 100%. CONCLUSIONS: The triage flowchart is able to accurately identify patients with CP not having an ACS. It may help triage nurses make quick decisions on who should be immediately seen and who could safely wait when delays in medical attention are unavoidable. Prospective validation is needed. PMID- 17920968 TI - Changes in the lipid class and proximate compositions of coho salmon (Oncorhynchus kisutch) smolts infected with the nematode parasite Philonema agubernaculum. AB - Coho salmon (Oncorhynchus kisutch) smolts infected with the nematode Philonema agubernaculum had 36% lower mean lipid content (1.4%) than nonparasitized coho salmon (2.2%) harvested simultaneously from the same outmigration. Lengths, weights, and condition factors, as well as protein and moisture content, did not differ significantly between the two groups. Lipid class compositions differed significantly between the parasitized and nonparasitized fish. None of the nematode-infected fish contained detectable triacylglycerols (TAG) or monoacylglycerols (MAG). In contrast, mean TAG and MAG contents of the nonparasitized fish totaled 5.5% of the extracted lipid. Infected smolts had lower cholesterol contents than did uninfected coho (17% for infected, 33% for uninfected). Parasitized fish had significantly higher levels of free fatty acids (mean of 57% for parasitized vs. 35% for nonparasitized) as well as the phospholipids phosphatidylethanolamine (PE) and phosphatidylcholine (PC). However, the PC/PE ratios for infected and noninfected coho did not differ significantly (2.2 for infected vs. 2.0 for uninfected). These differences suggest that the parasitic nematodes are either harvesting storage energy directly from the coho or are placing additional energetic demands on the fish to cope with the infection. PMID- 17920970 TI - Ability of myoglobin to predict mortality in patients admitted for exclusion of myocardial infarction. AB - BACKGROUND: Myoglobin can be used as an early marker to diagnose myocardial infarction (MI); and although nonspecific for myocardial necrosis, it seems to be a strong mortality predictor. Because myoglobin elevations are often present in patients with renal insufficiency, it is possible that the predictive value of myoglobin is secondary to identifying patients with renal insufficiency. METHODS: Consecutive patients admitted for MI exclusion without ST elevation on the initial electrocardiogram underwent serial assessment of cardiac markers (creatine kinase [CK], CK-myocardial band [MB], and troponin I [TnI]). Myoglobin was assessed at the time of admission and/or 3 hours later. Renal insufficiency was defined as a creatinine clearance <60 mL/min. Multivariate analysis was performed to identify predictors of 30-day and 1-year all-cause mortality. RESULTS: A total of 3461 patients were included in the analysis. Overall 30-day and 1-year mortality was 2.4% and 9.7%. Myoglobin was elevated in 675 (20%), CK MB in 421 (12%), and TnI in 517 (15%). Among the 993 patients with renal insufficiency, myoglobin was elevated in 43%, CK-MB in 17%, and TnI in 21%. Independent predictors of 30-day and 1-year mortality were similar and included age > or =65 years, prior MI, and an ischemic electrocardiogram, whereas myoglobin was the strongest multivariate predictor (odds ratio [OR] 2.8, 95% confidence interval [CI] 2.1-3.7), including those with renal insufficiency (OR 2.3, 95% CI 1.6-3.4). Troponin I had borderline predictive value (P = .08, OR 1.4, 95% CI 0.96-2.0), whereas CK-MB was not predictive in either group. CONCLUSIONS: Despite the absence of cardiac specificity, an elevated myoglobin strongly predicts mortality, even in patients with renal insufficiency. PMID- 17920971 TI - Risk factors associated with methicillin-resistant Staphylococcus aureus infection in patients admitted to the ED. AB - OBJECTIVES: The objective of our study was to define the characteristics of patients admitted to the emergency department (ED) presenting with a methicillin resistant Staphylococcus aureus (MRSA) infection. PATIENTS AND METHODS: The study included all patients admitted to the ED between January 2003 and December 2004 in whom a staphylococcal infection was documented. The risk factors associated with carriage of MRSA, the diagnosis made in the ED, and the treatment administered were established from the patients' medical files. The sites from which the bacteria were isolated, the spectrum of resistance of the staphylococci to different antibiotics, and the presence or absence of the gene coding for Panton-Valentin leukocidin for certain S aureus isolates were determined from the reports issued by the bacteriologic department. Two groups of patients were compared: those with an infection caused by MRSA and those with an infection due to methicillin-susceptible S aureus (MSSA). RESULTS: A total of 238 patients were included, 93 presenting with an infection caused by MRSA and 145 an infection due to MSSA. The patients harboring MRSA had a higher median age than those carrying MSSA (74 vs 61 years, P = .0001), experienced a greater loss of autonomy (according to the Knauss index), and had more comorbidity factors. Nine patients, younger than 40 years, presented with an infection due to MRSA in the absence of any comorbidity factor or any factor associated with carriage of these bacteria. Seven patients in the MRSA group were tested for Panton-Valentine leukocidin genes, and a positive result was obtained in 2 of them. Regardless of whether the infection was caused by MRSA or by MSSA, the bacteria were most frequently isolated from a cutaneous site, in 40% and 65% of the patients, respectively. Irrespective of the group, 28% of the patients presented with bacteremia. The spectrum of resistance of these MRSA strains suggested a hospital rather than community origin. The initial antibiotic therapy was rarely appropriate in the case of an infection due to MRSA. CONCLUSION: Infections caused by MRSA accounted for a high proportion of the staphylococcal infections diagnosed in the ED, necessitating a rational approach to the prescription of antibiotics for infections of this type. PMID- 17920972 TI - Prehospital analgesia with acupressure at the Baihui and Hegu points in patients with radial fractures: a prospective, randomized, double-blind trial. AB - BACKGROUND: Pain during transportation is a common phenomenon in emergency medicine. As acupressure has been deemed effective for pain management by the National Institutes of Health, we conducted a study to evaluate its effectiveness in prehospital patients with isolated distal radial fracture. METHODS: This was a prospective, randomized, double-blind study. Thirty-two patients were enrolled. Acupressure was performed either at "true" points or at "sham" points. Vital signs and pain and anxiety scores were recorded before and after the acupressure treatment. Normally distributed values were compared using the Student t test. RESULTS: Pretreatment scores for pain and anxiety were similar in the 2 groups (47.6 +/- 8.9 vs 51.2 +/- 8.7 visual analog scale [VAS] score for pain, 52.4 +/- 6.0 vs 47.5 +/- 9.3 VAS score for anxiety). At the hospital, patients in the true points group had significantly lower pain (36.6 +/- 11.0 vs 56.0 +/- 13.3 VAS score, P < .001) and anxiety scores (34.9 +/- 22.2 vs 53.4 +/- 19.7 VAS score, P = .022). CONCLUSION: Acupressure in the prehospital setting effectively reduces pain and anxiety in patients with distal radial trauma. PMID- 17920973 TI - Directed bedside transthoracic echocardiography: preferred cardiac window for left ventricular ejection fraction estimation in critically ill patients. AB - BACKGROUND: Bedside transthoracic echocardiography (TTE) performed by emergency physicians (EPs) is valuable in the rapid assessment and treatment of critically ill patients. We sought to determine the preferred cardiac window for left ventricular ejection fraction (LVEF) estimation by EP sonographers in a critically ill patient population. METHODS: Prospective investigator-blinded study of focused bedside TTE in a convenience sample of surgical intensive care patients. Investigators were faculty, fellows, or residents from an academic emergency medicine department. Five standard cardiac views were performed: parasternal long axis (PSLA), parasternal short axis (PSSA), subxiphoid 4 chamber, subxiphoid short axis, and apical 4-chamber (AFC). LVEF was determined using at least 1 cardiac view. Investigators rated their preference for each cardiac view on a 5-point Likert scale. RESULTS: A total of 70 studies were performed on 70 patients during a 6-month period. Users rated the PSLA as the most useful view for estimation of LVEF (mean 4.23; 95% confidence interval, 3.95 4.51). Pairwise comparisons of cardiac ultrasound views revealed PSLA was preferred over all other views (P < .05) except PSSA (P = .23). Complete 5 view examinations were not achieved in all patients (PSLA in 98%, PSSA in 96%, apical 4-chamber in 74%, subxiphoid 4-chamber in 35%, and subxiphoid short axis in 18%). Interobserver correlation of LVEF estimation was good (r = 0.86, r2 = 0.74, P < .0001). CONCLUSION: Parasternal long axis and PSSA are the preferred echocardiographic windows for EP estimation of LVEF using focused bedside TTE in critical care patients. This may be an important consideration in patients who often have physical barriers to optimal echocardiographic evaluation, are relatively immobile, and have unstable conditions requiring rapid assessment and intervention. PMID- 17920975 TI - Concordance of historical questions used in risk-stratifying patients with headache. AB - STUDY OBJECTIVE: We sought to determine whether the manner and order in which historical questions used to risk stratify for subarachnoid hemorrhage are asked significantly alter the response/concordance. METHODS: Adult patients complaining of headache in the emergency department were presented with 1 of 2 questionnaires each containing 2 variations of the pertinent question and differing only in their order. Data were primarily analyzed using the kappa statistic to determine whether rates of concordance are greater than would be expected by chance alone. And, as a secondary outcome, a sample of 120 was predetermined to be adequate to achieve 80% power in detecting a difference of 20% to 25% between questionnaires comparing the influence of order on concordance. RESULTS: The agreement corrected for chance for version 2, kappa = 0.51, is higher than the agreement corrected for chance for version 1, kappa = 0.28, a difference of 0.23 with a 95% confidence interval (-0.03 to 0.49; P = .08; SE, 0.13). The percentages of patients who answered the questions concordantly were 60% and 75%, respectively, for versions 1 and 2. The difference is 15% with a 95% confidence interval of ( 2% to 32%, P = .08). CONCLUSION: Although not statistically significant, our study indicates that up 38% answer these 2 very similar questions discordantly. Also, there appears to be a higher degree of concordance (15%) when patients are first asked, "When was the last time you had a headache this bad?" PMID- 17920974 TI - Women are less likely than men to receive prehospital analgesia for isolated extremity injuries. AB - OBJECTIVES: The goal of this study was to examine the effect of socioeconomic factors, such as ethnicity, income, age, and sex, on the administration of analgesia for isolated extremity injuries in the prehospital setting. METHODS: For this retrospective study, the electronic medical record of a large ground based emergency medical services agency was reviewed and all isolated extremity injuries occurring during the year 2005 were extracted. A total of 1009 cases met the inclusion criteria. Of these cases, 56 were excluded because of incomplete records, leaving 953 cases for analysis. Basic univariate analysis as well as logistic regression analysis were used to examine the relationship between analgesia administration and patient age, ethnicity, sex, income, subjective pain severity, and time under prehospital care. RESULTS: A total of 279 patients (29%) received morphine. Both univariate and logistic regression analysis revealed significant differences in analgesia administration based on sex (proportion of men receiving analgesia, 32.8%; women, 26.7%), initial pain severity, and time under prehospital care. Although no category of income was itself significant, a significant trend emerged in which increasing income was associated with increasing likelihood of receiving analgesia. There was no significant difference in analgesia based on patient age or ethnicity. CONCLUSION: This study suggests that women are less likely than men to receive prehospital analgesia for isolated extremity injuries. Patients with higher pain severity and longer duration of prehospital care are more likely to receive prehospital analgesia. Increasing levels of income were associated with increased rates of analgesia. The overall rate of prehospital analgesia administration for isolated extremity injuries in this population is higher than has been reported for other emergency medical services systems (29% vs 2%-18% in other recent studies), but there remains considerable room for improvement in the provision of prehospital analgesia. Further inquiry is needed to determine why certain populations such as women receive disproportionately less analgesia. PMID- 17920976 TI - A randomized controlled trial of intranasal fentanyl vs intravenous morphine for analgesia in the prehospital setting. AB - STUDY OBJECTIVE: The objective of the study was to compare intranasal fentanyl (INF) with intravenous morphine (IVM) for prehospital analgesia. METHODS: This was a randomized, controlled, open-label trial. Consecutive adult patients (n = 258) requiring analgesia (Verbal Rating Score [VRS] >2/10 noncardiac or >5/10 cardiac) were recruited. Patients received INF 180 mug +/- 2 doses of 60 mug at > or =5-minute intervals or IVM 2.5 to 5 mg +/- 2 doses of 2.5 to 5 mg at > or =5 minute intervals. The end point was the difference in baseline/destination VRS. RESULTS: Groups were equivalent (P = not significant) for baseline VRS [mean (SD): INF 8.3 (1.7), IVM 8.1 (1.6)] and minutes to destination [mean (SD): INF 27.2 (15.5), IVM 30.6 (19.1)]. Patients had a mean (95% confidence interval) VRS reduction as follows: INF 4.22 (3.74-4.71), IVM 3.57 (3.10-4.03); P = .08. Higher baseline VRS (P < .001), no methoxyflurane use (P < .01), and back pain (P = .02) predicted VRS reduction. Safety and acceptability were comparable. CONCLUSIONS: There was no significant difference in the effectiveness of INF and IVM for prehospital analgesia. PMID- 17920977 TI - An assessment of the association of bispectral index with 2 clinical sedation scales for monitoring depth of procedural sedation. AB - OBJECTIVE: We conducted a study to assess the correlation of bispectral index (BIS) to 2 clinical sedation scales. METHODS: This was a prospective, observational study. The BIS number was recorded at baseline and every 30 seconds. One investigator separately monitored the patients for depth of sedation using the Observer's Assessment of Alertness/Sedation and the Continuum of Depth of Sedation scales. RESULTS: During the 6-month period, 75 patients were enrolled. The Spearman correlation between the BIS and the Observer's Assessment of Alertness/Sedation was 0.59 (95% confidence interval [CI], 0.44-0.74). The Spearman correlation between the BIS and the Continuum of Depth of Sedation was 0.53 (95% CI, 0.36-0.70). The mean minimum BIS for patients without a complication was 70 (SD, 15.9) compared with 68 (SD, 12.9) for patients with a complication (difference = 2; 95% CI, -7-11). CONCLUSIONS: Our study demonstrated moderate correlation between BIS and the 2 clinical sedation scales. The correlation is not strong enough to be used reliably in a clinical setting. The mean minimum BIS scores were not significantly different for those with sedation complications vs those without complications. PMID- 17920978 TI - Emergency physicians do not use more resources to evaluate obese patients with acute abdominal pain. AB - OBJECTIVE: We hypothesized that emergency physicians would use more resources to evaluate acute abdominal pain in obese patients as compared with that in nonobese patients. METHODS: We conducted a secondary analysis of a prospective cohort of adults with acute abdominal pain. Collected data included self-reported height and weight, demographics, medical history, laboratory and x-ray results, and final diagnosis. We followed the patients until they obtained their final diagnosis or for up to 21 days. Patients were grouped according to their body mass index (BMI): nonobese (BMI < 30 kg/m2), obese (BMI = 30-40 kg/m2), and morbidly obese (BMI > 40 mg/m2). The main outcome measure was laboratory and radiographic testing. chi2 Tests and analysis of variance were used as appropriate. RESULTS: Of the 971 patients (mean age, 41 years; 62% black; 65% female), 665 (68%) were nonobese, 246 (25%) were obese, and 60 (6%) were morbidly obese. In comparing nonobese patients with obese patients, we found no difference in laboratory or radiographic testing (3.20 vs 3.21 tests; mean difference, 0.004; 95% confidence interval [CI], -0.26 to 0.27), physicians' pre-computed tomographic scan confidence level in their diagnosis (6.17 vs 6.04, mean difference, -0.13; 95% CI, -0.76 to 0.49), and emergency department (ED) length of stay (LOS; 7.40 vs 7.57 hours; mean difference, -0.17; 95% CI, -0.49 to 0.83). In comparing all 3 groups, we found no difference in diagnostic testing, ED LOS, surgical intervention (10% vs 5% vs 9%, P = .2), disposition, and final diagnosis (P > .05). CONCLUSIONS: Physicians do not use more resources to identify the etiology of acute abdominal pain in obese patients as compared with that in nonobese patients. Furthermore, ED LOS, likelihood of surgical intervention, physicians' confidence level in their preimaging diagnosis, and final diagnosis do not appear to be influenced by BMI. PMID- 17920979 TI - Bee swarmings in children. AB - Africanized honeybees (Apis mellifera scutellata) are now found in the southern and southwestern United States. Swarmings can result in hundreds to thousands of stings delivering a venom load capable of producing multisystem organ failure and death. The literature on mass envenomations is scarce, being limited to case reports and case series. There are no prospective studies on mass envenomations in children. METHODS: All patients were admitted to our toxicology service, and all stingers were counted. Laboratory data and clinical assessments were obtained at baseline, 8, and 16 hours after presentation. RESULTS: Nineteen patients with a median age of 3.6 years and a median of 2.64 stings per kilogram (range, 1-4.5) were enrolled. Fifteen children had vomiting. Only a mild increase in creatine kinase was seen. None developed coagulopathy or renal insufficiency. CONCLUSION: Envenomations of up to 4.5 stings per kilogram resulted in only mild systemic illness. Vomiting does not portend involvement of other organ systems. PMID- 17920980 TI - Can clear ascitic fluid appearance rule out spontaneous bacterial peritonitis? AB - OBJECTIVE: The objective of the study was to determine the sensitivity and specificity of abnormal ascitic fluid appearance in the detection of spontaneous bacterial peritonitis (SBP), as a sensitivity approaching 100% could exclude SBP if the fluid is clear. METHODS: We performed a retrospective review of all ascitic fluid samples sent for cell count over a 4-year period. Spontaneous bacterial peritonitis was defined as an absolute neutrophil count of more than 250 cells/mm3 or a positive culture. Abnormal fluid appearance was defined as hazy, cloudy, or bloody. RESULTS: There were 916 ascitic fluid samples analyzed during the study period. Eighteen percent of samples had a clear appearance, whereas 82% were abnormal. Abnormal ascitic fluid appearance had a sensitivity of 98.1% (95% confidence interval, 95.3%-99.5%) and a specificity of 22.7% (95% confidence interval, 19.4%-26.3%) in the detection of SBP. CONCLUSIONS: Abnormal fluid appearance has a high sensitivity for the detection of SBP. Thus, clear fluid might safely exclude SBP. PMID- 17920981 TI - Utility of Stewart's strong ion difference as a predictor of major injury after trauma in the ED. AB - INTRODUCTION: Base deficit (BD) is a validated surrogate for lactate in injured patients and correlates with trauma severity. Stewart proposed a more comprehensive measure of acidosis based on the strong ion difference (SID) (SID = Na + K + Mg + Ca - CL - lactate [mEq/L]). We compared operating characteristics of BD, anion gap (AG), and SID in identifying major injury in emergency department (ED) trauma patients. METHODS: This was a retrospective review. Major injury was defined as Injury Severity Score > or =15, blood transfusions, or significant drop in hematocrit. Receiver operating characteristic curves compared BD, AG, and SID in differentiating major from minor injuries. RESULTS: The study included 1181 patients. Both BD and SID were significantly (P = .0001) different after major vs minor injury (mean difference, 3.40; 95% confidence interval, 2.70 4.00 and mean difference, 2.50; 95% confidence interval, 1.90-3.10, respectively). Receiver operating characteristic curves were minimally different from one another (P = .0035). CONCLUSION: Stewart's SID can identify major injury in the ED. PMID- 17920982 TI - Impact of the 12-lead electrocardiogram on ED evaluation and management. AB - STUDY OBJECTIVE: This study was conducted to assess the impact (diagnostic, therapeutic, and disposition) of the 12-lead electrocardiogram (ECG) on emergency department (ED) patient evaluation and management. METHODS: This project was a prospective study of a convenience sample of 304 ED patients undergoing ECG analysis during their evaluation in the ED of a level 1 trauma center. The data collection for this study was divided into 4 parts. In part I, the treating physicians determined the specific reasons for ECG analysis; categories include complaint-based (eg, chest pain), syndrome-based (eg, overdose/poisoning), and system-based (eg, "requested by consult"). In part II, all treating physicians were surveyed before ECG interpretation regarding the future diagnostic, therapeutic, and disposition plans for the patient based only on history and physical examination. Their comments were recorded on a standardized data sheet. In part III, the physicians were surveyed after their interpretation of the ECG as to whether the results could suggest any further diagnostic information (eg, normal vs abnormal), or provide enough information for the patient care plan to be altered. In part IV of the study period, alterations to the original diagnostic, therapeutic, and disposition plans made by information provided by the ECG were obtained from the treating physician. RESULTS: A total of 304 patients underwent ECG examination and were used for data analysis. The average age of patients, of which 48% were men, was 60 years. The most common complaints that prompted electrocardiographic evaluation were chest pain and dyspnea. The most common reason an ECG was ordered was nursing staff protocol. Physicians determined that they were able to make a diagnosis based primarily on ECG in 33 (10.9%) cases. The total number of ECGs that were determined to be normal was 95 (31.3%), 7 (2.3%) of which allowed a rule-out diagnosis; 209 (68.7%) of total ECGs were determined to be abnormal, 72 (23.6%) of which were considered "of diagnostic significance." In 96 (31.6%) cases of electrocardiographic interpretation, alterations were made to the diagnostic, therapeutic, or disposition plans because of the information provided by the ECG. CONCLUSIONS: The ECG provides clinical information that frequently alters the management plan. PMID- 17920983 TI - Diagnosis and management of labile blood pressure during acute cerebrovascular accidents and other hypertensive crises. AB - It is estimated that with more than 40 million adults in the United States having uncontrolled hypertension, the risk of developing ischemic or hemorrhagic stroke in this population is significant. In addition, roughly 1 of 100 patients with essential hypertension will experience a hypertensive crisis during their lifetime, and these accelerated hypertensive emergencies and urgencies complicate more than 27% of all acute medical problems in patients presenting to emergency departments (EDs) in the United States. Arterial hypertension, a prominent feature of acute stroke syndrome, usually declines spontaneously within a few days, but its presence at hospital admission or its acute development during hospitalization is often associated with worsening stroke outcome and early mortality. Control of hypertension in patients with subarachnoid and intracerebral hemorrhage, both forms of acute stroke, is directed at maintaining adequate cerebral blood flow to minimize ischemic damage and control intracerebral pressure while reducing the risk of rebleeding and developing cerebrovasospasm. Inappropriate lowering of the blood pressure in acute stroke may increase neurologic damage. However, adequate blood flow around the central area of the acute ischemic stroke or penumbra may result in ischemic cells being salvaged. Clinicians must be mindful that accelerated hypertension is associated with other types of patients seen in the ED, such as perioperative patients and patients with traumatic head injuries. PMID- 17920984 TI - Ketamine for prehospital use: new look at an old drug. AB - INTRODUCTION: Ketamine has been used extensively for analgesia and anesthesia in many situations, including disaster surgery where extra personnel and advanced monitoring are not available. There are many features of ketamine that seem to make it an ideal drug for prehospital use. The reported use of ketamine in the prehospital environment is limited, however. The purpose of this study is to review the experience in the use of ketamine in a regional air ambulance service and suggest indications for its use in the prehospital setting. METHODS: This was a retrospective study of all patients transported by a regional aeromedical program. Patients were included in this study if the crew had used ketamine at any time during the flight. Data regarding the transport collected included patient age, type of transport, indications for ketamine use, and adverse reactions. RESULTS: During the period studied, ketamine was used in 40 patients. The age range was 2 months to 75 years. The indications and situations requiring use were varied and included both trauma and medical patients. Hypotension with need for analgesia, agitation or combativeness and intact airway, or pain unresponsive to narcotic medications were the most common indications for use. Ketamine was used both intravenous and intramuscular, even without intravenous access. There were no adverse reactions. CONCLUSIONS: Ketamine is an ideal drug for use in many prehospital situations. Our experience suggests that it is safe, effective, and may be more appropriate than drugs currently used by prehospital providers. PMID- 17920985 TI - Hyperthyroid cardiomyopathy is an infrequent but not exceptional cardiovascular manifestation of uncontrolled hyperthyroidism. PMID- 17920986 TI - Significance of comet tail artifacts at lung ultrasound. PMID- 17920987 TI - Isolated headache as the sole manifestation of dural sinus thrombosis. PMID- 17920988 TI - Ischemic stroke in a man with naphazoline abuse history. PMID- 17920989 TI - Transthoracic echocardiograph visualizing the efficiency of anticoagulant therapy for right atrium mobile thrombus in the elderly. PMID- 17920990 TI - Aneurysmal rebleeding episode after lumbar puncture. PMID- 17920991 TI - Isolated adrenal hemorrhage after minor blunt trauma. PMID- 17920992 TI - Anaphylactic reaction after a laboratory rat bite. PMID- 17920993 TI - Massive stroke in a previously healthy 7-year-old. PMID- 17920994 TI - Metabolism study in an 88-year-old woman with severe hypothermia during rewarming procedures. PMID- 17920995 TI - Use of the diffusion-weighted magnetic resonance imaging for early diagnosis of herpes simplex encephalitis in the ED: a case report. PMID- 17920996 TI - Aspiration pneumonitis and Newton's law of gravitation. PMID- 17920997 TI - Neurologic disease and vitamin B12 deficiency. PMID- 17920998 TI - Angioembolization as an effective alternative for hemostasis in intractable life threatening maxillofacial trauma hemorrhage: case study. PMID- 17921000 TI - [INTS: formation and political quality]. PMID- 17920999 TI - Post-transplant infections: single center experience from the developing world. AB - OBJECTIVE: To describe our experience of post-transplant infections in allogeneic stem cell transplants at the Armed Forces Bone Marrow Transplant Centre, Rawalpindi, Pakistan. METHODS: From July 2001 to September 2006, patients with malignant and non-malignant hematological disorders having human leukocyte antigen (HLA)-matched sibling donors were selected for transplant. Pre-transplant infection surveillance was carried out, and strict prophylaxis against infection was observed. After admission to the hospital, patients were kept in protective isolation rooms, equipped with a HEPA filter positive-pressure laminar airflow ventilation system. Bone marrow and/or peripheral blood stem cells were used as the stem cell source. Cyclosporin and prednisolone were used as prophylaxis against graft-versus-host disease (GVHD). The engraftment was monitored with cytogenetic/molecular analysis and change of blood group. Survival was calculated from the date of transplant to death or last follow-up. RESULTS: One hundred and fifty-four patients received allogeneic stem cell transplants from HLA-matched siblings for various hematological disorders at the Armed Forces Bone Marrow Transplant Centre, Rawalpindi, Pakistan between July 2001 and September 2006. Indications for transplant included aplastic anemia (n=66), beta-thalassemia major (n=40), chronic myeloid leukemia (n=33), acute leukemia (n=8), and miscellaneous disorders (n=7). One hundred and twenty patients were male and 34 were female. The median age of the patient cohort was 14 years (range 1 1/4-54 years). One hundred and thirty-six patients and 135 donors were cytomegalovirus (CMV) IgG-positive. One hundred and forty patients (90.9%) developed febrile episodes in different phases of post-transplant recovery. Infective organisms were isolated in 150 microbiological culture specimens out of 651 specimens from different sites of infections (23.0% culture positivity). Post-transplant infections were confirmed in 120 patients (77.9%) on the basis of clinical assessment and microbiological, virological, and histopathological examination. Mortality related to infections was 13.0%. Fatal infections included CMV disease (100% mortality, 6/6), disseminated aspergillosis (66.7% mortality, 4/6), pseudomonas septicemia (42.9% mortality, 9/21), and tuberculosis (25% mortality, 1/4). CONCLUSIONS: More than 90% of our patients developed febrile episodes with relatively low culture yield. The majority of infections were treated effectively, however CMV, aspergillosis, and pseudomonas infections remained problematic with high mortality. PMID- 17921004 TI - Management of pregnancy in a Jehovah's Witness. AB - In the successful management of a pregnant Jehovah's Witness, many issues must be addressed beyond those normally required for routine prenatal care. The clinician who undertakes such care should be well versed in the potential complications related to blood refusal, the antepartum management of anemia, and the intrapartum management of obstetric hemorrhage. Furthermore, these patients should be delivered in a tertiary care center because this increases their options for obtaining alternative management of hemorrhage. A woman who is well informed about her options can then decide exactly what she wants done in the event of a life-threatening obstetrical hemorrhage. PMID- 17921001 TI - [Evaluation of continuous education in transfusion for professionals in hospitals and clinics]. AB - Professional health workers need continuous update of knowledge in blood transfusion practice. To fulfill this expectation, the "Etablissement francais du sang (E.F.S) Auvergne-Loire", in cooperation with the Department of Regional Affairs in Health Services in the Auvergne region, has set up a proximity-based and a dedicated continuous education course, for more than three years. The content of this update course was based upon regulatory recommendations. The course is one day long; an evaluation of the course by the teachers as well as an evaluation of the teachers' level after the course is carried out. Each course attendant is given a CD-ROM which comprises all of the presentations of the day, along with relevant law texts. We now report on our experience in teaching 127 individual professionals in eight sessions: 95% gave full appreciation. This experience reached - up to now - 53% of the public and private hospitals with transfusion services in the Auvergne region and the Loire district and 90% of structures having remote blood banks. This experience can easily be extended to other regional E.F.S. settings. PMID- 17921005 TI - Intimate partner violence. AB - Intimate partner violence (IPV) has a lifetime prevalence of approximately 60% and is a leading cause of morbidity and mortality for women of all reproductive ages, especially among younger women and during pregnancy. Providers should recognize that every woman who has ever been partnered is at risk for IPV and should screen appropriately. When a woman screens positive for IPV, it important to consider the stages of change, to frame the response appropriately, to perform a risk assessment, to discuss interventions, and to document in the medical record accordingly. Screening has yet to translate into reduced rates of abuse, indicating that IPV is not simply a medical problem, but involves complex psychological, financial, familial, cultural, and legal issues. PMID- 17921006 TI - Approach to the acute abdomen in pregnancy. AB - Numerous physiologic changes in pregnancy may affect the presentation of abdominal pain in pregnancy. A high index of suspicion must be used when evaluating a pregnant patient with abdominal pain. General anesthesia is considered safe in pregnancy. Intraoperative monitoring and tocolytics should be individualized. Laparoscopic surgery should be performed in the second trimester when possible and appears as safe as laparotomy. If indicated, diagnostic imaging should not be withheld from the pregnant patient. Appendectomy and cholecystectomy appear to be safe in pregnancy. The reported incidence of adnexal masses and fibroids in pregnancy may increase with increasing use of first trimester ultrasound. Conservative management, with surgical management postpartum, appears reasonable in most cases. PMID- 17921007 TI - Current management of ectopic pregnancy. AB - Ectopic pregnancy continues to be one of the most common gynecologic emergencies and is the leading cause of pregnancy-related first-trimester death in the United States. The rate of ectopic pregnancy continues to rise because of increases in the incidences of its risk factors. However, improved modalities of early diagnosis and treatment have reduced both mortality and morbidity of this condition. In this article, the authors present an evidence-based review of the risk factors and presentation of ectopic pregnancy, including the utility of various diagnostic techniques, and compare the appropriateness and effectiveness of different therapeutic approaches. PMID- 17921008 TI - Postpartum hemorrhage. AB - Postpartum hemorrhage (PPH) is the leading cause of death related to pregnancy worldwide. Most deaths resulting from PPH are preventable. Physicians, nurses, midwives, and other birth attendants should be aware of the risk factors for PPH and be trained adequately in the preventive measures and management strategies for this pregnancy complication. Newer, less invasive technologies such as embolization may improve outcomes with PPH. Reducing the incidence of PPH and the mortality resulting from the condition should be a key goal of obstetrics services worldwide. This article focuses on the etiology, prediction, prevention, and management of PPH. PMID- 17921009 TI - Blood component therapy in obstetrics. AB - Hemorrhage is the leading cause of intensive care unit admission and one of the leading causes of death in the obstetric population. This emphasizes the importance of a working knowledge of the indications for and complications associated with blood product replacement in obstetric practice. This article provides current information regarding preparation for and administration of blood products, discusses alternatives to banked blood in the obstetric population, and introduces pharmacological strategies for treatment of hemorrhage. PMID- 17921010 TI - Early goal directed therapy for sepsis during pregnancy. AB - Sepsis is a leading cause of death in pregnancy and results in significant perinatal mortality. These deaths occur despite the younger age of pregnant patients, the low rate of comorbid conditions and the potential for effective interventions that should result in rapid resolution of illness. To date, no "evidence-based" recommendations are specific to the pregnant patient who is critically ill or septic. Optimal care for the septic patient requires a multidisciplinary team with expertise in obstetrics, maternal-fetal medicine, critical care, infectious disease, anesthesia, and pharmacy. Coordination of care and good communication amongst team members is essential. Incorporation of early goal directed therapy for suspected sepsis into obstetric practice is needed to optimize maternal and neonatal outcomes. PMID- 17921011 TI - Thromboembolism in pregnancy. AB - Venous thromboembolism in pregnancy is a clinical emergency that has been associated with significant risk for maternal and fetal morbidity and mortality. The adaptation of the maternal hemostatic system to pregnancy predisposes women to an increased risk of thromboembolism. A timely diagnosis of deep venous thrombosis is crucial because up to 24% of patients with untreated deep venous thrombosis develop a pulmonary embolism. Recent clinical guidelines identify compression venous ultrasound as the best way to diagnose deep venous thrombosis in pregnancy and CT pulmonary angiography as the best way to diagnose pulmonary embolism in pregnancy. Therapy involves supportive care and anticoagulation with unfractionated or low molecular weight heparin, depending on the clinical scenario. PMID- 17921012 TI - Shoulder dystocia: an update. AB - Shoulder dystocia has no consensus definition or management algorithm. Its incidence ranges from 0.2% to 3% and its occurrence is unpredictable. Risk factors for shoulder dystocia may include macrosomia, maternal diabetes, operative vaginal delivery, history of macrosomic infant or shoulder dystocia, labor abnormalities, post-term pregnancy, maternal obesity, advanced maternal age, fetal anthropometric variations, and male fetal gender. Once identified, multiple maneuvers can be applied in a stepwise fashion in an attempt to alleviate the dystocia. While training clinicians to manage shoulder dystocia is difficult because of its rare occurrence and lack of standardized management, all clinicians must be able to manage shoulder dystocia at any time. PMID- 17921013 TI - Diabetic ketoacidosis in pregnancy. AB - Episodes of diabetic ketoacidosis (DKA) can represent a life-threatening emergency for mother and fetus. The cornerstones of treatment of DKA are aggressive fluid replacement and insulin administration while ascertaining which precipitating factors brought about the current episode of DKA, and then treating accordingly to mitigate those factors. The incidence of DKA and factors unique to pregnancy are discussed in this article, along with the effects of the disease process on pregnancy. Clinical presentation, diagnosis, and treatment modalities are covered in detail to offer ideas to improve maternal and fetal outcome. PMID- 17921014 TI - Amniotic fluid embolism. AB - Amniotic fluid embolism is a catastrophic syndrome occurring during labor and delivery or immediately postpartum. Although presenting symptoms may vary, common clinical features include shortness of breath, altered mental status followed by sudden cardiovascular collapse, disseminated intravascular coagulation, and maternal death. It was first recognized as a syndrome in 1941, when two investigators described fetal mucin and squamous cells during postmortem examination of the pulmonary vasculature in women who had unexplained obstetric deaths. Since then, many studies, case reports, and series have been published in an attempt to elucidate the etiology, risk factors, and pathogenesis of this mysterious obstetric complication. PMID- 17921015 TI - Trauma in pregnancy. AB - Trauma is the leading nonobstetric cause of maternal mortality. The basic tenets of trauma evaluation and resuscitation should be applied in maternal trauma. Aggressive resuscitation of the mother is the best management for the fetus. Care must be taken to keep the patient in the left lateral decubitus position to avoid compression of the inferior vena cava and resultant hypotension. Radiographic studies should be used with care. Noninvasive diagnostics should be used when available. Cardiotocographic monitoring of a viable gestation should be initiated as soon as possible in the emergency department to evaluate fetal well-being. Urgent cesarean section should be considered if fetal distress is present or if the presence of the fetus is contributing to maternal instability. PMID- 17921016 TI - Cardiopulmonary resuscitation in pregnancy. AB - Although pregnancy and delivery in the United States are usually safe for mother and her newborn child, serious maternal complications, including cardiac arrest, can occur in the prenatal, intrapartum and postpartum periods. The clinical obstetrician can expect to encounter this complication in his or her career. The obstetrician must be aware of the special circumstances of resuscitation of the gravid woman to assist emergency medicine and critical care physicians in reviving the patient. Understanding the decision process leading to the performance of a perimortem cesarean and the actual performance of the cesarean delivery clearly are the responsibility of the obstetrician. PMID- 17921017 TI - Angiographic and interventional options in obstetric and gynecologic emergencies. AB - This article describes the role of angiographic and interventional techniques in the management of obstetrical and gynecologic hemorrhage. The complementary role of endovascular therapy is discussed and a review of management options for both peripartum hemorrhage and gynecologic hemorrhage is presented. The article describes special management options involving angiographic techniques for placentation abnormalities and arteriovenous malformations, and discusses a limited role for embolization in the management of ectopic pregnancies. The authors also present the outcomes of embolotherapy, associated complications, and implications for future fertility. This article describes the role of embolotherapy as it has evolved over the last three decades and clarifies the endovascular management options available to patients. PMID- 17921018 TI - Liability in high-risk obstetrics. AB - This article outlines the major causes of malpractice suits, focusing on those in obstetrical practice and reviewing the prime areas in antepartum and intrapartum care. Understanding the basic elements of medical malpractice allows a provider to better understand the nature of a suit for medical negligence. While the threat of a medical malpractice is ever present in obstetrics, practicing contemporary, evidence-based medicine with compassion and excellent communication is the best way to avoid alleged negligence. PMID- 17921019 TI - Dietary iron intake and serum ferritin in relation to 7.5 years structure and function of large arteries in the SUVIMAX cohort. AB - AIM: Few studies have investigated the relationship between iron stores and measures of atherosclerosis. Most of these studies were cross-sectional and yielded conflicting results. We aimed to assess the relationship between serum ferritin concentrations and dietary iron intake measured at baseline and 7.5 year pulse wave velocity (PWV), intima-media thickness (IMT) and plaques in a group of 824 men and women without known CVD, cancer or hemochromatosis. METHODS: The SUVIMAX study is a randomized double-blind, placebo-controlled primary prevention trial designed to test the effect of antioxidant supplementation in reducing ischemic cardiovascular diseases and cancer. RESULTS: In multivariate analyses, no association was found between baseline serum ferritin levels and IMT 7 years later (beta (95% CI)=0.003 (-0.005;0.011) in men; -0.005 (-0.013;0.004) and 0.001 (-0.011;0.009) in women, before and after menopause, respectively), plaques (OR (95% CI)=1.09 (0.88;1.34) in men; 0.93 (0.66;1.31) and 0.95 (0.70;1.29) in women, before and after menopause, respectively) or PWV (beta (95% CI)=0.078 ( 0.154;0.310) in men; -0.018 (-0266;0.231) in women before and after menopause). Results for dietary iron intake were similar. CONCLUSION: Our results do not support the hypothesis that dietary iron intake and body iron stores are deleterious to the structure and function of large arteries in subjects free of CVD, cancer or hemochromatosis. PMID- 17921020 TI - Multiple vertebral osteonecrosis. AB - Vertebral osteonecrosis classically presents with an intravertebral vacuum cleft phenomenon or a fluid-filled cleft on MR images. These clefts are usually found in older patients presenting with more severe fractures, more significant collapse and instability. Therefore, although considered for a long time as pathognomonic for vertebral osteonecrosis, vertebral clefts are now considered to represent fracture non-union. The double-line sign is classically described for osteonecrosis of long bones, but has been reported in one case of concurrent spinal cord and vertebral bone marrow radionecrosis. We present a case of a histologically confirmed multilevel vertebral osteonecrosis manifesting as a double-line sign in the absence of an associated vertebral collapse and unrelated to radiotherapy. PMID- 17921021 TI - Effectiveness of anti-IL1 in Schnitzler's syndrome. PMID- 17921023 TI - After bench to bedside: the impact of disease progression, commercial opportunities and available instrumentation on clinical outcome. PMID- 17921024 TI - Barriers to achieving commercial success for diagnostic and therapeutic radiopharmaceuticals. AB - The concluding session of this workshop focused on barriers that might be impeding the successful transition of radiopharmaceuticals from research tools to diagnostics and therapeutics that reach commercial success. Some lessons can be learned by reviewing the technology adoption life cycle as described by Geoffrey A. Moore (Moore, GA. Crossing the chasm. New York: HarperCollins, 2002). Additionally, a review of highly successful radiopharmaceuticals suggests that achieving commercial success may require advocacy by other interested groups. PMID- 17921022 TI - Medicoeconomic evaluation of hyaluronic acid for knee osteoarthritis in everyday practice: the MESSAGE study. AB - INTRODUCTION: Medicoeconomic data on treatments for osteoarthritis are scant. We investigated the impact of hyaluronic acid therapy on the cost of management of knee osteoarthritis. Our primary objective was to compare medical costs (admissions, outpatient visits, investigations, and treatments) and non-medical costs (sick leaves and transportation) from the perspective of the national health insurance system during the 3 months before and the 6 months after three intraarticular injections of hyaluronic acid. Our secondary objective was to evaluate treatment benefits in terms of pain, function, and quality of life. METHODS: Observational, multicenter, longitudinal, before-after study of the medical and economic effects of hyaluronic acid therapy for symptomatic knee osteoarthritis. RESULTS: Of the 296 assessable patients (mean age, 69 years; 30% with obesity; 65% women), only 5% of patients were withdrawn prematurely from the study. Significant improvements in the Lequesne index were found 3 and 6 months after treatment; the improvement was greater than 50% in over half the patients. Pain and quality-of-life scores improved significantly. Total cost of the disease decreased from 334 euros for the 3 pretreatment months to 295 euros and 233 euros for posttreatment months 1-3 and 4-6, respectively. CONCLUSION: The costs of knee osteoarthritis decreased during the 6 months after Suplasyn therapy, indicating that the cost of the medication was more than offset by the decreased need for other treatments. Concomitantly, clinical benefits were obtained. Under the conditions of everyday practice, hyaluronic acid may provide medical benefits at an acceptable cost. PMID- 17921025 TI - The future of SPECT in a time of PET. AB - As positron emission tomography (PET) imaging is becoming more prevalent in clinical practice, it is reasonable to ask if there will be a role for single photon emission computed tomography (SPECT) in the future. This article considers that question, focusing on areas where SPECT can differentiate itself from PET for fundamental reasons: breadth of available radionuclides, simultaneous imaging of multiple agents, cost-effectiveness and adaptability to specific imaging situations. The conclusion is that SPECT will continue to evolve and exist alongside PET and will grow the field of molecular imaging with improved efficiency and patient workflow. PMID- 17921026 TI - Clinical use of FDG PET. AB - INTRODUCTION: 2-[18F]-Fluoro-2-deoxy-D-glucose (FDG) has a wide range of possible clinical imaging applications in neurology, oncology and cardiology; however, there is also a large difference in the volume of actual clinical use between these applications. Three clinical questions are proposed and applied to the various clinical applications of FDG to understand this difference: (a) Are there competing tests? (b) Is there a cost-effective role in the clinical management? (c) Is the interpretation of the imaging study easy? METHODS: Each clinical application using FDG was reviewed with the three clinical questions proposed and the technical factor that may have contributed to addressing that question. The review of the application was based on the current practice of neurology, oncology and cardiology, and the contributing factors were based on the properties of FDG and the capabilities of the technology of current positron emission tomography scanners. RESULTS: A favorable answer was obtained for all three questions in oncology imaging with FDG. In cardiology and neurology, one of the three questions had an unfavorable outcome. CONCLUSION: Three clinical questions are proposed and applied to clinical FDG imaging in neurology, cardiology and oncology with the intent of better understanding the variation of clinical use. PMID- 17921027 TI - Simplifications in analyzing positron emission tomography data: effects on outcome measures. AB - Initial validation studies of new radiotracers generally involve kinetic models that require a measured arterial input function. This allows for the separation of tissue binding from delivery and blood flow effects. However, when using a tracer in a clinical setting, it is necessary to eliminate arterial blood sampling due to its invasiveness and the extra burden of counting and analyzing the blood samples for metabolites. In some cases, it may also be necessary to replace dynamic scanning with a shortened scanning period some time after tracer injection, as is done with FDG (F-18 fluorodeoxyglucose). These approximations represent loss of information. In this work, we considered several questions related to this: (1) Do differences in experimental conditions (drug treatments) or populations affect the input function, and what effect, if any, does this have on the final outcome measure? (2) How do errors in metabolite measurements enter into results? (3) What errors are incurred if the uptake ratio is used in place of the distribution volume ratio? (4) Is one- or two-point blood sampling any better for FDG data than the standardized uptake value? and (5) If blood sampling is necessary, what alternatives are there to arterial blood sampling? The first three questions were considered in terms of data from human dynamic positron emission tomography (PET) studies under conditions of baseline and drug pretreatment. Data from [11C]raclopride studies and those from the norepinephrine transporter tracer (S,S)-[11C]O-methyl reboxetine were used. Calculation of a metabolic rate for FDG using the operational equation requires a measured input function. We tested a procedure based on two blood samples to estimate the plasma integral and convolution that occur in the operational equation. There are some tracers for which blood sampling is necessary. Strategies for brain studies involve using the internal carotids in estimating the radioactivity after correcting for partial volume and spillover in order to eliminate arterial sampling. Some venous blood samples are still required for metabolite measurements. The ultimate solution to the problem of arterial sampling may be a wrist scanner, which acts as a small PET camera for imaging the arteries in the wrist. This is currently under development. PMID- 17921029 TI - Targeted alpha-particle radiotherapy with 211At-labeled monoclonal antibodies. AB - An attractive feature of targeted radionuclide therapy is the ability to select radionuclides and targeting vehicles with characteristics that are best suited for a particular clinical application. One combination that has been receiving increasing attention is the use of monoclonal antibodies (mAbs) specifically reactive to receptors and antigens that are expressed in tumor cells to selectively deliver the alpha-particle-emitting radiohalogen astatine-211 (211At) to malignant cell populations. Promising results have been obtained in preclinical models with multiple 211At-labeled mAbs; however, translation of the concept to the clinic has been slow. Impediments to this process include limited radionuclide availability, the need for suitable radiochemistry methods operant at high activity levels and lack of data concerning the toxicity of alpha particle emitters in humans. Nonetheless, two clinical trials have been initiated to date with 211At-labeled mAbs, and others are planned for the near future. PMID- 17921030 TI - Clinical acceptance of a molecular imaging agent: a long march with [99mTc]TRODAT. AB - In the past 10 years, significant progress has been made in using a technetium 99m dopamine transporter imaging agent, [99mTc]TRODAT, for routine clinical studies. Developing a molecular imaging agent from bench to the bedside is more than a simple scientific venture. Currently, Taiwan is the only place where [99mTc]TRODAT is approved for routine clinical use in the diagnosis of Parkinson's disease. The trials and tribulations of developing [99mTc]TRODAT for routine clinical use in Taiwan provide an interesting case study in how to (critics may say, how not to) develop a molecular imaging agent. PMID- 17921028 TI - Development of radioimmunotherapeutic and diagnostic antibodies: an inside-out view. AB - Only a handful of radiolabeled antibodies (Abs) have gained US Food and Drug Administration (FDA) approval for use in clinical oncology, including four immunodiagnostic agents and two targeted radioimmunotherapeutic agents. Despite the advent of nonimmunogenic Abs and the availability of a diverse library of radionuclides, progress beyond early Phase II radioimmunotherapy (RIT) studies in solid tumors has been marginal. Furthermore, [18F]fluorodeoxyglucose continues to dominate the molecular imaging domain, underscored by a decade-long absence of any newly approved Ab-based imaging agent (none since 1996). Why has the development of clinically successful Abs for RIT been limited to lymphoma? What obstacles must be overcome to allow the FDA approval of immuno-positron emission tomography (immuno-PET) imaging agents? How can we address the unique challenges that have thus far prevented the introduction of Ab-based imaging agents and therapeutics for solid tumors? Many poor decisions have been made regarding radiolabeled Abs, but useful insight can be gained from these mistakes. The following review addresses the physical, chemical, biological, clinical, regulatory and financial limitations that impede the progress of this increasingly important class of drugs. PMID- 17921031 TI - Human reporter genes: potential use in clinical studies. AB - The clinical application of positron-emission-tomography-based reporter gene imaging will expand over the next several years. The translation of reporter gene imaging technology into clinical applications is the focus of this review, with emphasis on the development and use of human reporter genes. Human reporter genes will play an increasingly more important role in this development, and it is likely that one or more reporter systems (human gene and complimentary radiopharmaceutical) will take leading roles. Three classes of human reporter genes are discussed and compared: receptors, transporters and enzymes. Examples of highly expressed cell membrane receptors include specific membrane somatostatin receptors (hSSTrs). The transporter group includes the sodium iodide symporter (hNIS) and the norepinephrine transporter (hNET). The endogenous enzyme classification includes human mitochondrial thymidine kinase 2 (hTK2). In addition, we also discuss the nonhuman dopamine 2 receptor and two viral reporter genes, the wild-type herpes simplex virus 1 thymidine kinase (HSV1-tk) gene and the HSV1-tk mutant (HSV1-sr39tk). Initial applications of reporter gene imaging in patients will be developed within two different clinical disciplines: (a) gene therapy and (b) adoptive cell-based therapies. These studies will benefit from the availability of efficient human reporter systems that can provide critical monitoring information for adenoviral-based, retroviral-based and lenteviral based gene therapies, oncolytic bacterial and viral therapies, and adoptive cell based therapies. Translational applications of noninvasive in vivo reporter gene imaging are likely to include: (a) quantitative monitoring of gene therapy vectors for targeting and transduction efficacy in clinical protocols by imaging the location, extent and duration of transgene expression; (b) monitoring of cell trafficking, targeting, replication and activation in adoptive T-cell and stem/progenitor cell therapies; (c) and assessments of endogenous molecular events using different inducible reporter gene imaging systems. PMID- 17921033 TI - Imaging multidrug resistance with 4-[18F]fluoropaclitaxel. AB - Multidrug resistance (MDR) is a cause of treatment failure in many cancer patients. MDR refers to a phenotype whereby a tumor is resistant to a large number of natural chemotherapeutic drugs. Having prior knowledge of the presence of such resistance would decrease morbidity from unsuccessful therapy and allow for the selection of individuals who may benefit from the coadministration of MDR inhibiting drugs. The Tc-99m-labeled single-photon-emitting radiotracers sestamibi and tetrofosmin have shown some predictive value. However, positron emitting radiotracers, which allow for dynamic quantitative imaging, hold promise for a more accurate and specific identification of MDRtumors.MDR-expressing tumors are resistant to paclitaxel, which is commonly used as a chemotherapeutic agent. 4-[18F]Fluoropaclitaxel (FPAC) is a PET-radiolabeled analogue of paclitaxel. Preclinical studies have shown the uptake of FPAC to be inversely proportional to tumor MDR expression. FPAC PET imaging in normal volunteers shows biodistribution to be similar to that in nonhuman primates. Imaging in a breast cancer patient showed FPAC localization in a primary tumor that responded to chemotherapy, while failure to localize in mediastinal disease corresponded with only partial response.FPAC PET imaging shows promise for the noninvasive pretreatment identification of MDR-expressing tumors. While much additional work is needed, this work represents a step toward image-guided personalized medicine. PMID- 17921035 TI - Sentinel lymph node mapping of breast cancer via intradermal administration of Lymphoseek. AB - Lymphoseek is a molecular imaging agent specifically designed for sentinel lymph node (SLN) mapping. We conducted a Phase I trial which measured the injection site clearance and SLN accumulation after a single intra dermal injection of Lymphoseek or [99mTc]sulfur colloid protocol. Ten patients with breast cancer participated in this study. Five patients received an intradermal administration of 1.0 nmol of 99mTc-labeled Lymphoseek and five patients received an intradermal administration of filtered [99mTc]sulfur colloid (fTcSC). Lymphoseek exhibited a significantly (P<.001) faster injection site clearance than fTcSC. The mean Lymphoseek clearance half-time was 2.61+/-0.72 h compared to 24.1+/-17.7 h for fTcSC. The mean SLN uptake of Lymphoseek (1.1+/-.5%) and fTcSC (2.5+/-4.9%) was statistically equivalent (P=.28). When an intra dermal injection was employed, Lymphoseek demonstrated faster injection site clearance than fTcSC. PMID- 17921034 TI - Targeting the treatment of drug abuse with molecular imaging. AB - Although imaging studies in and of themselves have significant contributions to the study of human behavior, imaging in drug abuse has a much broader agenda. Drugs of abuse bind to molecules in specific parts of the brain in order to produce their effects. Positron emission tomography (PET) provides a unique opportunity to track this process, capturing the kinetics with which an abused compound is transported to its site of action. The specific examples discussed here were chosen to illustrate how PET can be used to map the regional distribution and kinetics of compounds that may or may not have abuse liability. We also discussed some morphological and functional changes associated with drug abuse and different stages of recovery following abstinence. PET measurements of functional changes in the brain have also led to the development of several treatment strategies, one of which is discussed in detail here. Information such as this becomes more than a matter of academic interest. Such knowledge can provide the bases for anticipating which compounds may be abused and which may not. It can also be used to identify biological markers or changes in brain function that are associated with progression from drug use to drug abuse and also to stage the recovery process. This new knowledge can guide legislative initiatives on the optimal duration of mandatory treatment stays, promoting long lasting abstinence and greatly reducing the societal burden of drug abuse. Imaging can also give some insights into potential pharmacotherapeutic targets to manage the reinforcing effects of addictive compounds, as well as into protective strategies to minimize their toxic consequences. PMID- 17921032 TI - Impact of amyloid imaging on drug development in Alzheimer's disease. AB - Imaging agents capable of assessing amyloid-beta (Abeta) content in vivo in the brains of Alzheimer's disease (AD) subjects likely will be important as diagnostic agents to detect Abeta plaques in the brain as well as to help test the amyloid cascade hypothesis of AD and as an aid to assess the efficacy of anti amyloid therapeutics currently under development and in clinical trials. Positron emission tomography (PET) imaging studies of amyloid deposition in human subjects with several Abeta imaging agents are currently underway. We reported the first PET studies of the carbon 11-labeled thioflavin-T derivative Pittsburgh Compound B in 2004, and this work has subsequently been extended to include a variety of subject groups, including AD patients, mild cognitive impairment patients and healthy controls. The ability to quantify regional Abeta plaque load in the brains of living human subjects has provided a means to begin to apply this technology as a diagnostic agent to detect regional concentrations of Abeta plaques and as a surrogate marker of therapeutic efficacy in anti-amyloid drug trials. PMID- 17921036 TI - Platelet binding and biodistribution of [99mTc]rBitistatin in animal species and humans. AB - INTRODUCTION: 99mTc recombinant bitistatin (rBitistatin) is a radioligand for alphaIIbbeta3 (glycoproteins IIb/IIIa) receptor on platelets and is being developed as a diagnostic radiopharmaceutical for in vivo imaging of acute thrombi and emboli. Prior to the first administration of [99mTc]rBitistatin to human subjects, its biodistribution and effects on platelets were evaluated in animals. This paper reports findings in animal studies in comparison with initial findings in normal human subjects. METHODS: [99mTc]rBitistatin was administered to mice, guinea pigs and dogs to assess time-dependent organ distribution, urinary excretion and blood disappearance rates. Blood samples were analyzed to determine radioligand binding to circulating platelets and the extent of plasma protein binding. The effect of [99mTc]rBitistatin on circulating platelet count was determined. These factors were also determined in normal human subjects who received [99mTc]rBitistatin as part of a Phase I clinical trial. RESULTS: The main organs that accumulated [99mTc]rBitistatin were kidneys, liver and spleen in all animal species and humans. The main organs seen on human images were the kidneys and spleen. Liver uptake was fainter, and soft-tissue background was low. [99mTc]rBitistatin bound to circulating platelets in blood, with a higher percentage of binding to platelets in guinea pigs and dogs compared to that in humans. Plasma protein binding was low and of little consequence in view of platelet binding. The main route of excretion was through the urine. [99mTc]rBitistatin did not affect platelet counts in humans or dogs. CONCLUSIONS: [99mTc]rBitistatin, when administered at low doses for imaging, has no adverse effects on platelets and has the qualitative biodistribution predicted by animal studies. [99mTc]rBitistatin was found to bind to circulating platelets in humans, suggesting that it will be able to bind to activated platelets in vivo in patients with acute thrombi. PMID- 17921037 TI - Serotonin-1A receptor imaging in recurrent depression: replication and literature review. AB - INTRODUCTION: Serotonin-1A receptor (5-HT1AR) function appears to be decreased in major depressive disorder (MDD) based on physiological responses to 5-HT1AR agonists in vivo and to 5-HT1AR binding in brain tissues postmortem or antemortem. We have previously assessed 5-HT1AR binding potential (BP) in depression using positron emission tomography (PET) and [carbonyl-(11)C]WAY 100635, and we have demonstrated reduced 5-HT1AR BP in the mesiotemporal cortex (MTC) and raphe in depressives with primary recurrent familial mood disorders (n=12) versus controls (n=8) [Drevets WC, Frank E, Price JC, Kupfer DJ, Holt D, Greer PJ, Huang Y, Gautier C, Mathis C. PET imaging of serotonin 1A receptor binding in depression. Biol Psychiatry 1999;46(10):1375-87]. These findings were replicated by some, but not other, studies performed in depressed samples that were more generally selected using criteria for MDD. In the current study, we attempted to replicate our previous findings in an independent sample of subjects selected according to the criteria for primary recurrent depression applied in our prior study. METHODS: Using PET and [carbonyl-(11)C]WAY-100635, 5-HT1AR BP was assessed in 16 depressed subjects and 8 healthy controls. RESULTS: Mean 5 HT1AR BP was reduced by 26% in the MTC (P<.005) and by 43% in the raphe (P<.001) in depressives versus controls. CONCLUSIONS: These data replicate our original findings, which showed that BP was reduced by 27% in the MTC (P<.025) and by 42% in the raphe (P<.02) in depression. The magnitudes of these reductions in 5-HT1AR binding were similar to those found postmortem in 5-HT1AR mRNA concentrations in the hippocampus in MDD [Lopez JF, Chalmers DT, Little KY, Watson SJ. Regulation of serotonin 1A, glucocorticoid, and mineralocorticoid receptor in rat and human hippocampus: implications for neurobiology of depression. Biol Psychiatry 1998;43:547-73] and in 5-HT1AR-binding capacity in the raphe in depressed suicide victims [Arango V, Underwood MD, Boldrini M, Tamir H, Kassir SA, Hsiung S, Chen JJ, Mann JJ. Serotonin 1A receptors, serotonin transporter binding and serotonin transporter mRNA expression in the brainstem of depressed suicide victims. Neuropsychopharmacology 2001;25(6):892-903]. There exists disagreement within the literature, however, regarding the presence and direction of 5-HT1AR-binding abnormalities in depression, which may be explained in some cases by differences in anatomical location (e.g., [Stockmeier CA, Shapiro LA, Dilley GE, Kolli TN, Friedman L, Rajkowska G. Increase in serotonin-1A autoreceptors in the midbrain of suicide victims with major depression--postmortem evidence for decreased serotonin activity. J Neurosci 1998;18(18):7394-401]) and in other cases by pathophysiological heterogeneity within MDD (e.g., some depressives hypersecrete cortisol, which would be expected to down-regulate 5-HT1AR expression [Lopez JF, Chalmers DT, Little KY, Watson SJ. Regulation of serotonin 1A, glucocorticoid, and mineralocorticoid receptor in rat and human hippocampus: implications for neurobiology of depression. Biol Psychiatry 1998;43:547-73]). Antidepressant drug treatment does not alter these abnormalities in 5-HT1AR binding [Sargent PA, Kjaer KH, Bench CJ, Rabiner EA, Messa C, Meyer J, Gunn RN, Grasby PM, Cowen PJ. Brain serotonin1A receptor binding measured by positron emission tomography with [11C]WAY-100635: effects of depression and antidepressant treatment. Arch Gen Psychiatry 2000;57(2):174-80; Moses-Kolko EL, Price JC, Thase ME, Meltzer CC, Kupfer DJ, Mathis CA, Bogers WD, Berman SR, Houck PR, Schneider TN, Drevets WC. Measurement of 5-HT1A receptor binding in depressed adults before and after antidepressant drug treatment using positron emission tomography and [11C]WAY 100635. Synapse 2007;61(7):523-30] but may compensate for blunted 5-HT1AR function by increasing post-synaptic 5-HT1AR transmission [Chaput Y, de Montigny C, Blier P. Presynaptic and postsynaptic modifications of the serotonin system by long-term administration of antidepressant treatments. An in vivo electrophysiologic study in the rat. Neuropsychopharmacology 1991;5(4):219-29]. PMID- 17921038 TI - Challenges in clinical studies with multiple imaging probes. AB - This article addresses two related issues: (a) When a new imaging agent is proposed, how does the imager integrate it with other biomarkers, either sampled or imaged? (b) When we have multiple imaging agents, is the information additive or duplicative and how is this objectively determined? Molecular biology is leading to new treatment options with reduced normal tissue toxicity, and imaging should have a role in objectively evaluating new treatments. There are two roles for molecular characterization of disease. Molecular imaging measurements before therapy help predict the aggressiveness of disease and identify therapeutic targets and, therefore, help choose the optimal therapy for an individual. Measurements of specific biochemical processes made during or after therapy should be sensitive measures of tumor response. The rules of evidence are not fully developed for the prognostic role of imaging biomarkers, but the potential of molecular imaging provides compelling motivation to push forward with convincing validation studies. New imaging procedures need to be characterized for their effectiveness under realistic clinical conditions to improve the management of patients and achieve a better outcome. The purpose of this article is to promote a critical discussion within the molecular imaging community because our future value to the overall biomedical community will be in supporting better treatment outcomes rather than in detection. PMID- 17921039 TI - [A voluminous sacrococcygeal teratoma: prenatal diagnosis, in-utero treatment and obstetric management]. AB - Sacrococcygeal teratoma is the most common and benign fetal tumor. Fetuses with sacrococcygeal tumors that are predominantly solid and highly vascularized have a high risk of fatal issue. Hydrops and tumor hemorrhage are associated with a highest risk of fetal death. Management of these tumors includes ultrasound scan with Doppler and magnetic resonance imaging (MRI) is usually used for evaluation of its intrapelvic extension and relationship to the other structures. New in utero treatments as vascular coagulation have been applied in fetuses with highly vascular teratomas but these techniques are still experimental and need more investigations. The management of delivery depends on associated anomalies, tumor vascularity and size. PMID- 17921040 TI - Effects of ultrasound on apoptosis induced by anti-CD20 antibody in CD20-positive B lymphoma cells. AB - AIM: The present study was conducted to examine the thermal and non-thermal effects of ultrasound on apoptosis induced by anti-CD20 monoclonal antibody (rituximab). MATERIALS AND METHODS: SU-DHL-4 cells, a CD20-positive cell line derived from B cell lymphomas with a BCL2 gene rearrangement, were exposed to continuous 1 MHz ultrasound for therapeutic use under an air- or CO(2)-saturated condition to control cavitation. Early apoptosis (EA) and secondary necrosis (SN) were examined by flow cytometry. Cavitation was determined by detecting the hydroxyl radicals derived from pyrolysis of water molecules using electron paramagnetic resonance-spin trapping. To assess thermal effects, cells were treated in a temperature-controlled water bath. RESULTS: There was a significant additive increase in EA and EA+SN observed in cells treated with rituximab combined with heat at 42 degrees C or non-thermal ultrasound at 0.5 W/cm(2) under an air-saturated condition, where heat or ultrasound induced some cell death. A significant synergistic increase in EA and EA+SN was observed in cells treated with rituximab and ultrasound at 2.5 W/cm(2) under CO(2)-saturated conditions, where inertial cavitations were completely suppressed. No enhancement was observed at a temperature less than 40 degrees C or ultrasound at 0.5 W/cm(2) under CO(2)-saturated conditions. CONCLUSION: These results suggest that the immuno-therapeutic application of ultrasound at relatively high-intensities combined with rituximab thus produces synergistic effects under conditions where the non-thermal and non-cavitational effects are predominant. PMID- 17921042 TI - Learning multiple layers of representation. AB - To achieve its impressive performance in tasks such as speech perception or object recognition, the brain extracts multiple levels of representation from the sensory input. Backpropagation was the first computationally efficient model of how neural networks could learn multiple layers of representation, but it required labeled training data and it did not work well in deep networks. The limitations of backpropagation learning can now be overcome by using multilayer neural networks that contain top-down connections and training them to generate sensory data rather than to classify it. Learning multilayer generative models might seem difficult, but a recent discovery makes it easy to learn nonlinear distributed representations one layer at a time. PMID- 17921041 TI - Iron homeostasis and toxicity in retinal degeneration. AB - Iron is essential for many metabolic processes but can also cause damage. As a potent generator of hydroxyl radical, the most reactive of the free radicals, iron can cause considerable oxidative stress. Since iron is absorbed through diet but not excreted except through menstruation, total body iron levels buildup with age. Macular iron levels increase with age, in both men and women. This iron has the potential to contribute to retinal degeneration. Here we present an overview of the evidence suggesting that iron may contribute to retinal degenerations. Intraocular iron foreign bodies cause retinal degeneration. Retinal iron buildup resulting from hereditary iron homeostasis disorders aceruloplasminemia, Friedreich's ataxia, and panthothenate kinase-associated neurodegeneration cause retinal degeneration. Mice with targeted mutation of the iron exporter ceruloplasmin have age-dependent retinal iron overload and a resulting retinal degeneration with features of age-related macular degeneration (AMD). Post mortem retinas from patients with AMD have more iron and the iron carrier transferrin than age-matched controls. Over the past 10 years much has been learned about the intricate network of proteins involved in iron handling. Many of these, including transferrin, transferrin receptor, divalent metal transporter-1, ferritin, ferroportin, ceruloplasmin, hephaestin, iron-regulatory protein, and histocompatibility leukocyte antigen class I-like protein involved in iron homeostasis (HFE) have been found in the retina. Some of these proteins have been found in the cornea and lens as well. Levels of the iron carrier transferrin are high in the aqueous and vitreous humors. The functions of these proteins in other tissues, combined with studies on cultured ocular tissues, genetically engineered mice, and eye exams on patients with hereditary iron diseases provide clues regarding their ocular functions. Iron may play a role in a broad range of ocular diseases, including glaucoma, cataract, AMD, and conditions causing intraocular hemorrhage. While iron deficiency must be prevented, the therapeutic potential of limiting iron-induced ocular oxidative damage is high. Systemic, local, or topical iron chelation with an expanding repertoire of drugs has clinical potential. PMID- 17921045 TI - Endings in the middle: current knowledge of interstitial telomeric sequences. AB - Interstitial telomeric sequences (ITSs) consist of tandem repeats of the canonical telomeric repeat and are common in mammals. They are localized at intrachromosomal sites, including those repeats located close to the centromeres and those found at interstitial sites, i.e., between the centromeres and the telomeres. ITSs might originate from ancestral intrachromosomal rearrangements (inversions and fusions), from differential crossing-over or from the repair of double-strand break during evolution. Three classes of ITSs have been described in the human genome, namely, short ITSs, long subtelomeric ITSs and fusion ITSs. The fourth class of ITSs, pericentromeric ITSs, has been found in other species. The function of ITSs can be inferred from the association of heritable diseases with ITS polymorphic variants, both in copy number and sequence. This is one of the most attractive aspects of ITS studies because it leads to new and useful markers for genetic linkage studies, forensic applications, and detection of genetic instability in tumors. Some ITSs also might be hotspots of chromosome breakage, rearrangement and amplification sites, based on the type of clastogens and the nature of ITSs. This study will contribute new knowledge with respect to ITSs' biology and mechanism, prevalence of diseases, risk evaluation and prevention of related diseases, thus facilitates the design of early detection markers for diseases caused by genomic instability. PMID- 17921044 TI - Antibiotic resistant Staphylococcus aureus: a paradigm of adaptive power. AB - Nothing documents better the spectacular adaptive capacity of Staphylococcus aureus than the response of this important human and animal pathogen to the introduction of antimicrobial agents into the clinical environment. The effectiveness of penicillin introduced in the early 1940s was virtually annulled within a decade because of the plasmid epidemics that spread the ss-lactamase gene through the entire species of S. aureus. In 1960 within one to two years of the introduction of penicillinase resistant ss-lactams (methicillin), methicillin resistant S. aureus (MRSA) strains were identified in clinical specimens. By the 1980s, epidemic clones of MRSA acquired multidrug resistant traits and spread worldwide to become one of the most important causative agents of hospital acquired infections. In the early 2000s, MRSA strains carrying the Tn1546 transposon-based enterococcal vancomycin resistant mechanism were identified in clinical specimens, bringing the specter of a totally resistant bacterial pathogen closer to reality. Then, in the late 1990s, just as effective hygienic and antibiotic use policies managed to bring down the frequency of MRSA in hospitals of several countries, MRSA strains began to show up in the community. PMID- 17921043 TI - BH3 mimetics to improve cancer therapy; mechanisms and examples. AB - Tumor cell survival is highly dependent on the expression of certain pro-survival Bcl-2 family proteins. An attractive therapeutic approach is to inhibit these proteins using agents that mimic the Bcl-2 homology 3 (BH3) domains of the proapoptotic Bcl-2 family members, which neutralize these proteins by binding to their surface hydrophobic grooves. A number of BH3 mimetic peptides and small molecules have been described, a few of which have advanced into clinical trials. Recent studies have highlighted ABT-737, a bona fide BH3 mimetic and potent inhibitor of antiapoptotic Bcl-2 family members, as a promising anticancer agent. This review summarizes recent advances in understanding the mechanisms of action of BH3 domains and several classes of BH3 mimetics, as well as the prospects of using these agents to improve cancer therapy. PMID- 17921046 TI - Nuclear architecture, chromosome domains and genetic damage. AB - The interphase nucleus structure is radially organised into three-dimensional discrete chromosome territories or domains (CTs) surrounded by a channel network named the interchromatin compartment (IC) which harbours factors involved in DNA replication or repair as well as RNA transcription and processing. Gene-rich chromosomes are centrally located whereas gene-poor ones are bound to the nuclear outskirts. Chromatin dynamics also reflect nuclear compartment organisation. Replication timing and topology as well as active or inactive chromatin residence are highly regulated in eukaryotic cells. Early replicating euchromatin, high transcription levels and histone H4 hyperacetylation (H4+a) characterise the nuclear interior while late replicating heterochromatin, poor transcription rates and underacetylated histone H4 distinguish the nuclear periphery. Active chromatin loops mostly map to the surface of CTs and protrude into the IC whereas inactive loops mainly reside in the CTs core. Response of nuclear compartments to clastogen insult in terms of chromosomal aberrations is not uniform. The euchromatic, H4+a nuclear interior seems more sensitive to ionising radiation, nucleases and chemical agents. Topological changes of CTs occur after induced radiation damage. Chromatin remodeling associated to DNA synthesis, CTs relative positioning, loci spatial proximity, intermingling of chromatin loops and transcriptional activity could be critical to determine chromosome damage localisation, genomic instability and cancer-prone translocation frequencies. PMID- 17921047 TI - Transmission of Dientamoeba fragilis: evaluation of the role of Enterobius vermicularis. AB - The role of Enterobius vermicularis in the transmission of Dientamoeba fragilis has been evaluated in two groups of patients admitted to the Parasitology Laboratory of Celal Bayar University: one group with E. vermicularis infection (n=187, Pinworm Group), and the other with D. fragilis infection (n=126, Dientamoeba Group). The presence of the other parasite, pinworm or Dientamoeba, was investigated with the microscopic examination of cellophane tape and stool samples for three consecutive days. In the Pinworm Group, 9.6% of the patients were found to be coinfected with D. fragilis, while 25.4% of the patients in the Dientamoeba Group were found to be coinfected with pinworms. The coincidence rates of D. fragilis and E. vermicularis, higher than the prevalence of each parasite in similar populations, suggest a common relation between these two parasites, possibly in entering the human body. E. vermicularis infection was found to be significantly more common in younger children (p<0.001), indicating that younger children may also be at higher risk for D. fragilis infection. These findings also raise the question of whether the unrelated symptoms of the pinworm infected patients such as abdominal pain and diarrhea may actually be due to overlooked Dientamoeba infections. PMID- 17921048 TI - A quantitative approach of using genetic algorithm in designing a probability scoring system of an adverse drug reaction assessment system. AB - BACKGROUND: The detrimental effects of adverse drug reactions (ADRs) are well established. Hence, precise and accurate assessment of ADRs' causality which can differentiate signal from noise is crucial in screening, management and minimisation of ADRs. OBJECTIVE: The current study reported our attempt to improve the scoring system of a previously published algorithm of ADR assessment by our group using a genetic algorithm approach so that the final score can measure the probability of ADR causality. DESIGN: Using ADR cases obtained from the Centre for Drug Administration, the national centre for pharmacovigilance in Singapore, with known causality probability values as reference points, rules were developed to define possible combinations of criteria for 'Definite' ADR cases and 'Probable' ADR cases. A new scoring system was developed using these parameters with the help of genetic algorithm, and tested on 37 'Definite' and 431 'Not Definite' ADR cases. In addition, sensitivity and specificity analysis were performed to allow a comparison of performance between our algorithm and that used by the Adverse Drug Reaction Advisory Committee in Australia (ADRAC). RESULTS: The new scoring system is able to provide a probability of the causality of an ADR by a suspected drug. When applied to the 'Definite' and 'Not Definite' ADR reports, the new algorithm gave a sensitivity of 83.8% and specificity of 71.0%. CONCLUSIONS: Using a quantitative method of assessing causality in the new algorithm allows rare and new ADRs to be more readily identified since a quantitative score can give a more precise degree of ADR causality. This scoring system that provides a probability score would help to make this algorithm more informative and assistive for clinicians, regulatory agencies or pharmaceutical companies to generate ADR alerts. The higher sensitivity value displayed by our algorithm also shows that it would be a good ADR screening tool. PMID- 17921049 TI - Long-term beta-adrenergic stimulation leads to downregulation of protein phosphatase inhibitor-1 in the heart. AB - Desensitization of the beta-adrenoceptor/cAMP/PKA pathway is a hallmark of heart failure. Inhibitor-1 (I-1) acts as a conditional amplifier of beta-adrenergic signalling downstream of PKA by inhibiting type-1 phosphatases in the PKA phosphorylated form. I-1 is downregulated in failing hearts and thus presumably contributes to beta-adrenergic desensitization. To test whether I-1 downregulation is a consequence of excessive adrenergic drive in heart failure, rats were treated via minipumps with isoprenaline 2.4 mg/kg/day (ISO) or 0.9% NaCl for 4 days. As expected, chronic ISO increased heart-to-body weight ratio by approximately 40% and abolished the inotropic response to acute ISO in papillary muscles by approximately 50%. In the ISO-treated hearts I-1 mRNA and protein levels were decreased by 30% and 54%, respectively. This was accompanied by decreased phospholamban phosphorylation (-40%), a downstream target of I-1, and a reduction in 45Ca2+ uptake (-54%) in membrane vesicles. Notably, phospholamban phosphorylation correlated significantly with I-1 protein levels indicating a causal relationship. We conclude that I-1 downregulation in heart failure is likely a consequence of the increased sympathetic adrenergic drive and participates in desensitization of the beta-adrenergic signalling cascade. PMID- 17921050 TI - Tobacco smoke-induced left ventricular remodelling is not associated with metalloproteinase-2 or -9 activation. AB - AIM: To investigate the role of MMP-2 and MMP-9 in cardiac remodelling induced by tobacco smoke exposure in rats. METHODS: Rats were allocated into two groups: C (n=9): control animals; ETS (n=9): exposed to tobacco smoke. After 4months, the animals underwent echocardiography, morphometric study and determination of MMP-2 and MMP-9 activity. RESULTS: ETS rats had larger diastolic (C=15.6 +/-1.2 mm/kg, ETS=18.0+/-0.9 mm/kg; p < 0.001) and systolic (C=7.3+/-1.2 mm/kg, ETS=9.2+/-0.9 mm/kg; p=0.001) ventricular diameters adjusted for body weight. Fractional shortening (C=53+/-4.8%, ETS=48+/- 3.3%; p=0.031) and ejection fraction (C=0.89+/ 0.03, ETS=0.86+/-0.02; p=0.030) were smaller in the ETS group. Myocyte cross sectional area (C=245+/-8 microm2, ETS=253+/-8 microm2; p=0.028) was higher in ETS rats. There were no differences in MMP-2 (C=50+/-14%; ETS=43+/-11%, p=0.228) or MMP-9 (C=0.36+/-0.3%; ETS=0.62+/-0.3%, p=0.630) activity between the groups. CONCLUSION: MMP-2 and MMP-9 did not participate in the remodelling process induced by tobacco smoke exposure. PMID- 17921051 TI - Effect of tolterodine on sleep structure modulated by CYP2D6 genotype. AB - OBJECTIVE: Tolterodine, a drug for the treatment of overactive bladder symptoms, has a limited entry into the brain, which makes cognitive side effects seldom. However, some case reports have described central-nervous side effects such as sleepiness. The aim of this retrospective analysis was to investigate whether tolterodine-related effects on sleep stage parameters could be explained by different CYP2D6 metabolizer characteristics of subjects. METHODS: Data were taken from two randomized, double-blind, placebo-controlled studies conducted in a cross-over design. Forty-eight volunteers underwent 4 two-night attended polysomnographic studies. Subjective quality of sleep and cognitive function were assessed. A single dose of 4 mg tolterodine or placebo was administered before sleep. Forty-four volunteers gave informed consent for genotyping. We found 19 extensive metabolizers (EM), 20 intermediate metabolizers (IM), 4 poor metabolizers (PM) and 1 ultrarapid metabolizer. There were no significant differences between the groups regarding demographic data. RESULTS: Rapid eye movement (REM) sleep duration as a percentage of total sleep time showed significant reduction (p=0.019) in the group carrying one or more deficient alleles (IM+PM). No significant difference was found with two active alleles of CYP2D6 in the EM group. REM latencies under tolterodine displayed a tendency towards prolongation, which was irrespective of the metabolizer status. Subjective sleep parameters did not show statistically significant changes after tolterodine. Cognitive skills were not affected. CONCLUSION: Our retrospective analysis reveals that a decrease of REM sleep under tolterodine is found only in individuals carrying one or two deficient CYP2D6 alleles. PMID- 17921052 TI - Decrease in myocardial 123I-MIBG radioactivity in REM sleep behavior disorder: two patients with different clinical progression. AB - Although decrease in myocardial iodine-123 metaiodobenzylguanidine ((123)I-MIBG) radioactivity has been reported in patients with rapid eye movement (REM) sleep behavior disorder (RBD), its pathophysiology has not been thoroughly disclosed. We report two RBD patients with differing clinical progression, in whom myocardial (123)I-MIBG scintigraphy was performed. One 69-year-old patient had more than a 20-year history of idiopathic RBD and showed a decrease in myocardial (123)I-MIBG radioactivity. The other 69-year-old patient started to manifest nocturnal behaviors at age 62, then mild parkinsonism at age 68, and showed a similar decrease in myocardial (123)I-MIBG radioactivity both before and after the onset of parkinsonism. These cases suggest that RBD could develop in diverse patterns of clinical progression even if signs of underlying Lewy body pathology are uniformly indicated. PMID- 17921053 TI - Confirmatory factor analysis of the Epworth Sleepiness Scale (ESS) in patients with obstructive sleep apnoea. AB - BACKGROUND: The Epworth Sleepiness Scale (ESS [Johns MW. A new method for measuring daytime sleepiness: the Epworth sleepiness scale. Sleep 1991;14(6):540 5]) has been used frequently to assess daytime sleepiness, particularly in the context of clinical sleep disorders. Its psychometric properties are still unclear, particularly when used to evaluate sleep propensity in patients with obstructive sleep apnoea. METHODS: The present study used confirmatory factor analysis (CFA) to investigate a potential single-factor structure of the ESS in a sample of 759 Australian patients with a diagnosis of obstructive sleep apnoea by the treating physicians. RESULTS: CFA results from showed that the original single-factor structure proposed by Johns [Johns MW. Reliability and factor analysis of the Epworth Sleepiness Scale. Sleep 1992;15(4):376-81] did not adequately fit the data. A re-specified single-factor solution provided a good fit for data, and this improved fit was confirmed on a second CFA. CONCLUSIONS: The findings suggest that standard scoring of the ESS should be interpreted cautiously for patients with obstructive sleep apnoea. PMID- 17921054 TI - Residual subjective daytime sleepiness under CPAP treatment in initially somnolent apnea patients: a pilot study using data mining methods. AB - BACKGROUND AND PURPOSE: Despite correct treatment with positive airway pressure (PAP), obstructive sleep apnea (OSA) patients sometimes remain subjectively somnolent. The reliability of the Epworth Sleepiness Scale (ESS) has been established for healthy subjects and patients under stable conditions; the ESS may eventually vary among treated OSA patients, biasing the results of a cross sectional analysis of persisting sleepiness. The objective of this study was to depict the evolution of subjective vigilance under treatment using an index of ESS variability (DeltaESS). METHODS: In 80 OSA patients (apnea-hypopnea index [AHI]=54+/-26/h), initially somnolent (ESS=15+/-3) and treated with auto titrating PAP (APAP) (oxyhaemoglobin desaturation index 3% [ODIapap]=3.4+/-2.2/h; daily APAP use=5.3+/-1.5 h) during 434+/-73 days, ESS scores were regularly collected four times every 109+/-36 days. DESS was calculated and data mining methods (Segmentation and Decision Tree) were used to determine homogeneous groups according to the evolution of ESS scores. RESULTS: When assessed cross sectionally, 14-25% of the subjects were recognized as somnolent, depending on the moment when ESS was administered. Using data mining methods, three groups were clearly identifiable: two without residual somnolence - group 1, n=38 (47%), with high DeltaESS=-2.9+/-0.8, baseline ESS=16.3+/-3.3, AHI=58.5+/-26.1/h, mean ESSapap=5.1+/-2.4 and group 2, n=31 (39%), with low DeltaESS=-1.1+/-0.5, baseline ESS=13.2+/-1.4, AHI=53+/-27.3/h, mean ESSapap=8.8+/-1.9; and one with persisting sleepiness; group 3, n=11 (14%), with low DeltaESS=-0.3+/-0.8, baseline ESS=16.3+/-3, AHI=38.7+/-10.8/h, mean ESSapap=14.1+/-1.9. Compliance to PAP was high and comparable in the three groups. Age and body mass index (BMI) did not differ. CONCLUSION: Data mining methods helped to identify 14% of subjects with persisting sleepiness. Validation needs to be done on a larger population in order to determine predictive rules. PMID- 17921055 TI - Relationships between headache and sleep in a non-clinical population of children and adolescents. AB - BACKGROUND: Headache and sleep are related in different ways and alterations of chronobiological mechanisms are involved in headache. We investigated the relationships between headache and sleep quality in a large non-clinical population of children and adolescents and evaluated the relationship between headache and circadian typologies. METHODS: A total of 1073 children and adolescents (50.9% males; mean age=10.56; range=8-15 years) were recruited from four schools in Rome. They filled out the questionnaires individually in classrooms, after brief group instruction about answer formats. The questionnaires included (a) a self-report headache questionnaire to collect information on different aspects of headache attacks based on the International Classification of Headache Disorders-2nd edition (ICHD-2); (b) the School Sleep Habits Survey that incorporated questions about sleep habits, the Sleep-Wake Problems Behaviour Scale (SWPBS), the Sleepiness Scale (SLS) and the Morningness/Eveningness Questionnaire (MEQ). RESULTS: According to ICHD-2 criteria, we classified 70 (6.5%) children as Migraine Group (MG), 135 (12.7%) as Non-Migraine Headache Group (NMG), and the remaining 868 (80.8%) were classified as Headache-Free Group (HFG). No clear differences have been found between MG and NMG regarding the frequency of the attacks, although MG showed a significantly increased frequency of long-lasting attacks. The modality of onset of pain and the location of pain was similar in both groups. The most frequent triggering factor for headache in MG and NMG was "a bad sleep" (32.2%) followed by emotional distress (27.8%). No differences have been found between MG, NMG and HFG in sleep schedule or sleep duration. MG and NMG showed significantly higher scores on the SWPBS vs. HFG, while MG presented higher scores on the SLS compared to NMG and HFG. MG presented lower MEQ scores, indicating a more pronounced eveningness. CONCLUSIONS: The relationships between headache and sleep problems are evident even in a non-clinical population of children and adolescents, with MG showing poorer sleep quality, sleepiness and a tendency toward eveningness. PMID- 17921056 TI - Perceived severity of restless legs syndrome across the female life cycle. AB - BACKGROUND: As with migraine, female sex hormones may explain the high prevalence of restless legs syndrome (RLS) in women and therefore influence RLS severity across the female life cycle. OBJECTIVE AND METHODS: To test this hypothesis, we performed a questionnaire-based transversal survey in female members of the French Association of patients with RLS. Five hundred thirty-six women fulfilled the RLS criteria and completed the International RLS Severity Scale (IRLSSS) and questionnaire about reproductive behaviour, RLS history and perception of RLS symptom severity during pregnancy, menses and menopause. RESULTS: Patients with at least one child showed a significantly higher mean IRLSSS score than women without children and 23% of the patients declared having perceived worsening of symptoms during pregnancy. Perceived RLS severity was increased during menses in 29% of non-menopaused patients and 69% of the patients reported worsening of symptoms following menopause. In these patients, a tendency towards higher IRLSSS scores was noted. Regression analysis revealed a correlation between higher IRLSSS scores and an early age at onset of RLS. CONCLUSIONS: Female hormonal changes do not account for the variation in perceived severity in women with RLS during their hormonal milestones and their role in the pathophysiology of RLS is unlikely. PMID- 17921057 TI - Unilateral periodic leg movements during wakefulness and sleep after a parietal hemorrhage. PMID- 17921060 TI - Half of patients with obstructive sleep apnea have a higher NREM AHI than REM AHI. PMID- 17921061 TI - Prevalence of recurrent otitis media in habitually snoring school-aged children. AB - INTRODUCTION: The pathophysiology of obstructive sleep apnea (OSA) and recurrent otitis media (ROM) is intimately associated with the presence of adenotonsillar hypertrophy in children. However, it remains unclear whether habitually snoring children have a higher prevalence of ROM and whether they require tympanostomy tube placement more frequently. METHODS: Questionnaires collected from parental surveys of 5- to 7-year-old children attending the public schools in Louisville, KY were retrospectively reviewed for the presence of habitual snoring (HS), ROM, and the need for tympanostomy tube insertion. RESULTS: There were 16,321 surveys with complete datasets (51.2% boys; 18.6% African American (AA) with a mean age of 6.2+/-0.7 years). Of these children, 1844 (11.3%) were HS (53% boys; 25.9% AA); and, of these, 827 HS had also a positive history of ROM (44.8%) with a slight predominance in males (55%). In addition, 636 of these children underwent placement of tympanostomy tubes (i.e., 34.4% of all HS and 76.9% of ROM). Among the 14,477 non-snoring children (NS), ROM was reported in 4247 NS children (29.3%; p<0.000001; odds ratio [OR]: 1.95; confidence interval [CI]: 1.77-2.16) of which 57.6% were boys, and 1969 NS with ROM underwent tympanostomy tube placement (i.e., 46.3% of those with ROM and 13.6% of all non-snoring children). Thus, the risk for tympanostomy tube placement was also greater among HS compared to NS children (p<0.00001; OR: 2.19; CI: 1.98-2.43). CONCLUSIONS: Habitual snoring is associated with a significant increase in the prevalence of recurrent otitis media and the need for tympanostomy tube placement. Further studies aiming to assess the prevalence of obstructive sleep apnea among children with ROM are needed. PMID- 17921062 TI - Association between sleep duration and hemoglobin A1c level. AB - OBJECTIVE: The association between sleep habits and hemoglobin A1c (HbA1c) level has not been sufficiently examined. In the present study of residents in a local community, the associations between sleep duration and HbA1c level were examined. METHODS: Self-administered questionnaires were mailed to 1062 residents in a rural community in Japan, and completed questionnaires were collected. At the time of collection, the fasting plasma glucose and Hb(1c levels were measured using peripheral blood samples. For the analyses, values that were considered to represent high levels were a fasting plasma glucose level of 126mg/dl and a HbA1c level of 6.5%. Logistic regression analyses were performed to examine the associations between sleep duration and high fasting plasma glucose or high HbA1c levels. RESULTS: The prevalence of high fasting plasma glucose and high HbA1c levels was significantly high (p<0.01) in subjects with a short or a long sleep duration. Logistic regression analyses demonstrated a significant association between high HbA1c level and sleep duration. The adjusted odds ratios for a high HbA1c level showed high values with regard to both short and long sleep durations. CONCLUSIONS: HbA1c level showed a U-shaped association with sleep duration. These results suggest that there may be an appropriate range of sleep duration in individuals with glucose tolerance disorders. It is expected that the present findings will contribute to the treatment and prevention of diabetes mellitus. PMID- 17921063 TI - Central apneas in a case of Crisponi syndrome. PMID- 17921064 TI - Titration procedures for nasal CPAP: automatic CPAP or prediction formula? AB - BACKGROUND: The best method for titration of continuous positive airway pressure (CPAP) therapy in obstructive sleep apnea (OSA) syndrome has not yet been established. The 90th or 95th percentiles of the pressure titrated over time by automatic CPAP (A-CPAP) have been recommended as reference for prescribing therapeutic fixed CPAP (F-CPAP). We compared A-CPAP to F-CPAP, which was determined by a common prediction formula. METHODS: Forty-five patients who were habituated to F-CPAP underwent titration polysomnography. In a double-blind, randomized order, each patient used an A-CPAP device in the autotitration and in the fixed pressure mode during one half of the night. Apnea-hypopnea index (AHI) and pressure profiles were primary outcomes. Bias and precision were additionally assessed for both CPAP modes. RESULTS: No significant differences in various sleep parameters or in subjective sleep quality evaluation were found. The AHI was effectively lowered in both CPAP modes (A-CPAP 7.7 [10.8]events/h versus F CPAP 5.4 [9.0]events/h, p=0.061). Comparison of group means showed that F-CPAP closely paralleled mean (Pmean) and median (P50), but not the 95th percentile (P95) pressure, of A-CPAP. While bias was lowest for Pmean and P50, there was a lack of precision in all A-CPAP pressure categories. CONCLUSIONS: We confirm that F-CPAP set by prediction formula is not worse in terms of AHI control than A CPAP. On average, F-CPAP parallels Pmean and P50 but not P95. However, due to imprecise matching, individual F-CPAP values cannot be derived from Pmean or P50. PMID- 17921065 TI - Augmentation in restless legs syndrome is associated with low ferritin. AB - BACKGROUND AND PURPOSE: Augmentation is a major problem with dopaminergic therapy for restless legs syndrome (RLS), and predictors of augmentation have not yet been identified. We aimed to analyze the relationship between baseline ferritin level and occurrence of augmentation in a retrospective analysis of a prospective double-blind trial of cabergoline versus levodopa on augmentation in RLS. PATIENTS AND METHODS: Patients who experienced augmentation were compared to patients who did not experience augmentation. RESULTS: Augmentation symptoms causing premature discontinuation from the study or which were tolerated (n=36, ferritin: 85+59 ng/ml) were associated with lower levels of serum ferritin compared to patients without augmentation (n=302, ferritin: 118+108 ng/ml, p=0.0062). CONCLUSIONS: Ferritin as a marker of iron storage may play an important role in the pathophysiology of RLS and may prove to be a biomarker for the development of augmentation under dopaminergic therapy. PMID- 17921066 TI - Partial sleep deprivation: impact on the architecture and quality of sleep. AB - OBJECTIVE: The study was aimed at the examination of the acute and cumulative impact of partial sleep deprivation (PSD) on architecture and quality of sleep, on circadian rhythm and on daytime fatigue. METHODS: Time in bed for 16 healthy male volunteers was reduced from 8 to 5h during four consecutive nights, followed by two recovery nights. This scheme matches the terms of service and sleep rhythms of rescue helicopter pilots in Germany. Polysomnography was recorded during each night, and motor activity and body core temperature were recorded continuously. Subjective sleep quality and alertness were acquired by questionnaires. RESULTS: In the course of PSD, electroencephalography (EEG) showed a reduction of S1 and S2 (p=0.039), whereas S3, S4 (0.024) and rapid eye movement (REM; p=0.030) sleep increased. Subjective sleep quality improved, while sleep need (p<0.001) and fatigue (p<0.001) deepened. These effects declined rapidly after one recovery night. CONCLUSIONS: PSD alters sleep and daytime alertness. The results indicate that not only slow wave sleep (SWS) but also REM is important for sleep. Sleep became more effective when wake periods and sleep latencies decreased, thereby improving subjective sleep quality. Sleepiness, sleep need and fatigue increased immediately and accumulated throughout the restricting procedure. PMID- 17921067 TI - Increased plasma levels of inflammatory markers and upper airway resistance during sleep in pre-eclampsia. AB - OBJECTIVES: To evaluate pregnancy-associated sleep disorders, pregnancy outcomes and inflammatory markers in pre-eclampsia and normal pregnancy (control). PATIENTS AND METHODS: We studied 15 consecutive pre-eclamptic women and 14 controls. Pre-eclamptic women underwent overnight pulse oximetry and nasal pressure measurements at a university teaching hospital, and the sleep study for the controls was performed at home. Mean gestation was 31 weeks. Nasal airflow was carefully analyzed visually, and the time with flow limitation was calculated as a percentage of total recording time. At the time of the sleep study, the subjects were clinically evaluated, they answered sleep questionnaires, and fasting blood samples were drawn for tumor necrosis factor alpha TNF-alpha, interleukin 6 (IL-6) and sensitive C-reactive protein. Pregnancy outcomes were collected after delivery. RESULTS: Pre-eclampsia patients spent significantly more time with flow limitation (mean+/-SD: 21+/-18% vs. 4+/-9%), had higher plasma levels of TNF-alpha (6.2+/-2.3 ng/l vs. 4.1+/-ng/l) and IL-6 (4.4+/-ng/l vs. 1.2+/-0.4 ng/l), had more generalized edema, had increased fatigue and snoring, and had poorer pregnancy outcomes than did controls. Age, gestational age, mean SpO2 and body mass index did not differ between the groups. CONCLUSIONS: Pregnant women with pre-eclampsia showed significantly more nasal flow limitation during the night, higher fasting IL-6 and TNF-alpha plasma levels, more edema and worse pregnancy outcomes than did healthy pregnant women. PMID- 17921068 TI - Familial concordance in cancer survival: a Swedish population-based study. AB - BACKGROUND: Nowadays, the fact that cancers can aggregate in families is generally accepted. The aim of this study was to complete a comprehensive analysis of cancer-survival concordance in parents and their children diagnosed with the same cancer. METHODS: We used a population-based Swedish family database, that included about three million families and data for more than a million individuals with cancer. We analysed survival in children in relation to parental survival by use of the Kaplan-Meier method. We then modelled the risk in children in relation to parental survival by use of two multivariate proportional hazard (Cox) models adjusting for possible confounders of survival. FINDINGS: In our univariate Kaplan-Meier analysis, children with the same cancer as their parent and whose parent had died within 10 years of diagnosis showed significantly worse survival for breast (log rank p=0.01), colorectal (p=0.04), and prostate cancer (p=0.05) than those whose parents were alive at 10 years from diagnosis. By use of Cox modelling, we noted an increased hazard ratio for death from cancer in children with poor parental survival compared with those with good parental survival for colorectal cancer (hazard ratio [HR] 1.44 [95% CI 1.01 2.01]), lung cancer (1.39 [1.00-1.94]), breast cancer (1.75 [1.13-2.71]), ovarian cancer (2.23 [0.78-6.34]), and prostate cancer (2.07 [1.13-3.79]). All hazard ratio estimates, except for ovarian cancer, were significant, with significant trends of increasing risk of death in children by degree of worsening survival outcome in parents defined in quartiles of survival (ie, good [best quartile], expected [middle two quartiles], or poor [worst quartile]). INTERPRETATION: Our findings suggest that cancer-specific survival in parents predicts survival from the same cancer in their children. Consequently, data on survival in a parent might have the potential to guide treatment decisions and genetic counselling. Finally, molecular studies to highlight the genetic determinants of cancer survival are now warranted. PMID- 17921070 TI - Early detection of right ventricular systolic dysfunction by using myocardial acceleration during isovolumic contraction in patients with mitral stenosis. AB - AIM: The aim of the study was to determine if the tissue Doppler imaging (TDI) derived myocardial acceleration during isovolumic contraction (IVA) of tricuspid lateral annulus could be used in early detection of RV systolic dysfunction in patients with mitral stenosis (MS), before the clinical signs of systemic venous congestion occur. METHODS: One hundred and twelve patients with rheumatic MS without relevant regurgitation and 60 control subjects were enrolled in the study. Conventional echocardiographic parameters (mitral valve area, transmitral diastolic gradients, pulmonary artery pressure, RV fractional shortening, pulmonary flow acceleration time, tricuspid annular plane systolic excursion) and TDI-derived systolic velocities of tricuspid annulus (isovolumic myocardial acceleration: IVA, peak myocardial velocity during isovolumic contraction: IVV, peak systolic velocity during ejection period: Sa and RV Tei index) were recorded from all patients. RESULTS: TDI-derived IVA, IVV, Sa and Tei index were found to be significantly decreased in patients with MS. IVA was the only parameter which had a significant negative correlation with the traditional echocardiographic parameters and RV Tei index in patients with MS. Additionally, in subgroup analyses, IVA was significantly lower in patients with severe degree of MS. CONCLUSION: TDI-derived right ventricular IVA may be used as an adjunctive, reliable, noninvasive parameter for the early detection of right ventricular systolic dysfunction in patients with MS but without signs of systemic venous congestion. PMID- 17921069 TI - Gene expression profiles of bronchoalveolar cells in pulmonary TB. AB - The host response to Mycobacterium tuberculosis includes macrophage activation, inflammation with increased immune effector cells, tissue necrosis, and cavity formation, and fibrosis, distortion, and bronchiectasis. To evaluate the molecular basis of the immune response in the lungs of patients with active pulmonary tuberculosis (TB), we used bronchoalveolar lavage to obtain cells at the site of infection. Affymetrix GeneChip microarrays and cDNA nylon filter microarrays interrogated gene expression in bronchoalveolar lavage (BAL) cells from 11 healthy controls and 17 patients with active pulmonary TB. We found altered gene expression for 69 genes in TB versus normal controls that included cell surface markers, cytokines, chemokines, receptors, transcription factors, and complement components. In addition, TB BAL cell gene expression patterns segregated into 2 groups: one suggestive of a T helper type 1 (Th1) cellular immune response with increased signal transducer and activator of transcription-4 (STAT-4), interferon-gamma (IFN-gamma receptor), and monokine induced by IFN gamma (MIG) expression with increased IFN-gamma protein levels in BAL fluid; the other group displayed characteristics of Th2 immunity with increased STAT-6, CD81, and IL-10 receptor expression. We were able to demonstrate that a Th2 presentation could change to a Th1 pattern after anti-tuberculous treatment in 1 TB patient studied serially. These gene expression data support the conclusion that pulmonary TB produces a global change in the BAL cell transcriptome with manifestations of either Th1 or Th2 immunity. PMID- 17921071 TI - A new ELISA for the three-spined stickleback (Gasterosteus aculeatus L.) spiggin, using antibodies against synthetic peptide. AB - The aim of this study was to develop an enzyme-linked immunosorbent (ELISA) assay to quantify spiggin in the three-spined stickleback. Spiggin is a glue protein produced in the kidney of male three-spined stickleback under the control of androgens during the breeding period. Disturbances of spiggin production in male fish and abnormal induction of spiggin in female fish are considered as valuable biomarkers of exposure to (anti-)androgenic chemicals. Polyclonal antibodies against a peptide sequence of spiggin (HRD-16) were used and the specificity of the antibodies was verified by Western blotting and direct ELISA experiments. By using HRD-16 antibodies and spiggin standard preparation, a competitive ELISA was set-up and validated. This assay appears sensitive, with a detection limit of 0.5 U/mL, and specific, as shown by the competition curves, obtained by serial dilution of male and female kidney homogenates, that were parallel to the spiggin standard curves. The ability of the spiggin ELISA to quantify spiggin induction was achieved by exposing male and female three-spined sticklebacks to 0.1 and 1 microg/L of methyltestosterone. The results show a significant dose-dependent induction of spiggin in methyltestosterone-exposed female fish compared to controls. PMID- 17921072 TI - Taxonomy-based partitioning of the Gene Ontology. AB - The Gene Ontology (GO) project is a collaborative effort to construct ontologies which facilitate biologically meaningful annotation of gene products. In some situations, only a generic or a species-specific subset of all GO terms is required to annotate and analyze the results of a particular biomedical experiment. We show that by defining explicit links between terms in the GO and terms in the Taxonomy of Species (TS) it is possible to automatically create partitions of the GO according to various taxonomic criteria. Our framework is based on three logically defined relations--validity, specificity, and relevance- used to link terms in the Gene Ontology with terms in the Taxonomy. The major advantages of this approach, as compared to the traditional GO slims methodology, are: unambiguous semantics of GO-TS annotations, significant reduction of the effort needed to manually select GO terms appropriate for a particular taxonomic context, ability to generate views of the GO even for taxa for which no explicit links with GO terms exist, logical consistency of such views, and automated updates of TS-dependent GO subsets. Incorporation of the proposed framework into the GO may improve the usability of the ontology for those scientists who focus their research on a particular species or a specific class of organisms. PMID- 17921073 TI - Correlating viral phenotypes with phylogeny: accounting for phylogenetic uncertainty. AB - Many recent studies have sought to quantify the degree to which viral phenotypic characters (such as epidemiological risk group, geographic location, cell tropism, drug resistance state, etc.) are correlated with shared ancestry, as represented by a viral phylogenetic tree. Here, we present a new Bayesian Markov Chain Monte Carlo approach to the investigation of such phylogeny-trait correlations. This method accounts for uncertainty arising from phylogenetic error and provides a statistical significance test of the null hypothesis that traits are associated randomly with phylogeny tips. We perform extensive simulations to explore and compare the behaviour of three statistics of phylogeny trait correlation. Finally, we re-analyse two existing published data sets as case studies. Our framework aims to provide an improvement over existing methods for this problem. PMID- 17921074 TI - Determination of meloxicam in human plasma using a HPLC method with UV detection and its application to a pharmacokinetic study. AB - A simple and sensitive high performance liquid chromatography method using UV detection (HPLC-UV) for the determination of meloxicam in human plasma was developed and validated. After extraction with diethyl ether, the chromatographic separation of meloxicam was carried out using a reverse phase Sunfire C18 column (150 mm x 4.6mm, 5 microm) with a mobile phase of acetonitrile-20mM potassium hydrogen phosphate (40:60, v/v, pH 3.5) and UV detection at a wavelength of 355 nm. The flow rate of mobile phase was 1.2 ml/min and the retention time of meloxicam and internal standard, piroxicam, was found to be 11.6 and 6.3 min, respectively. The calibration curve was linear within the concentration range, 10 2400 ng/ml (r2>0.9999). The lower limit of quantification was 10 ng/ml. This method improved the sensitivity for the quantification of meloxicam in plasma using a HPLC-UV. The mean accuracy was 98-114%. The coefficient of variation (precision) in the intra- and inter-day validation was 1.6-4.3 and 2.4-7.3%, respectively. The pharmacokinetics of meloxicam was evaluated after administering an oral dose of 15 mg to 11 healthy Korean subjects. The AUCinf, Cmax, tmax and t1/2 were 42.4+/-13.2 microg h/ml, 1445.7+/-305.5 ng/ml, 4.1+/-0.3h and 22.0+/ 4.9h, respectively. PMID- 17921075 TI - Out-of-pocket health care expenditures due to excess of body weight in Portugal. AB - The prevalence for excessive weight has also been increasing dramatically in Portugal over the last decades. The consequences for families as well as for the publicly funded Portuguese health care system are a matter of policy interest. This paper uses an econometric model to compute the fraction of the national out of-pocket health care expenditures attributable to overweight and obesity among Portuguese adults. Given that public health care system pays for a substantial share of the national health care expenditures, the estimated the out-of-pocket expenditures is only a share of the total expenditures. Per-capita expenditures and the burden that obesity and overweight impose on families are also estimated. Two waves of the Portuguese National Health Survey (NHS), namely; 1995/1996 and 1998/1999 are considered. The results suggest that out-of-pocket expenditures due to excess weight have increased sharply during these 3 years. The two-part model estimates suggest that the obese and overweight are more likely to incur out-of pocket health care expenditures but, in the restricted sample of those that incur expenditures, there is weak or no evidence that the obese or overweight spend, on average, more than those of normal weight. Overall, it is estimated that in 1995/1996, more than 1.8% out-of-pocket health care expenditures were attributable to obesity and 2% to overweight (although not statistically significant). The estimated percentages are over 2.9% for obesity and 4% for overweight in 1998/1999. Combined, the estimated attributable percentage of national out-of-pocket expenditures due to excess weight was 3.8% in 1995/1996 and 6.9% in 1998/1999. Per-capita expenditures due to obesity or overweight are small, on average, in absolute terms, but they can be a significant cost for low income families. With respect to public policy concerns, the results suggest that measures which only slightly increase the out-of-pocket health care expenditures of being obese (overweight) are likely to be inefficient. PMID- 17921076 TI - Improvement of mechanical and biological properties of porous CaSiO3 scaffolds by poly(D,L-lactic acid) modification. AB - Porous calcium silicates (CaSiO3, WT) are regarded as a potential bioactive material for bone tissue engineering. However, their insufficient mechanical strength and high dissolution (degradation) limit their biological applications. The aim of this study is to surface modify WT scaffolds with poly(d,l-lactic acid) (PDLLA) to improve their mechanical and biological properties. The phase composition, microstructure, porosity and interconnectivity of WT and PDLLA modified WT (WTPL) scaffolds were analyzed by X-ray diffraction, scanning electron microscopy and micro-computerized tomography. The WTPL scaffolds maintained a more uniform and continuous inner network, compared to that of the WT scaffolds, while maintaining the pore size, porosity and interconnectivity of the original materials. The compressive strength, compressive modulus and percentage strain of the WT and WTPL scaffolds were assessed in air and phosphate buffered saline. PDLLA modification significantly improved the compressive strength and decreased the brittleness of the WT scaffolds. The weight loss and apatite-forming ability of the two scaffolds were evaluated by soaking them in simulated body fluid (SBF) for 1, 3, 7, 14 and 28days. PDLLA modification decreased the dissolution of the WT scaffolds while maintaining their apatite forming ability in SBF. In addition, PDLLA modification improved the spreading and viability of human bone-derived cells. Our results indicate that PDLLA modified CaSiO3 scaffolds possess improved mechanical and biological properties, suggesting their potential application for bone tissue regeneration. PMID- 17921077 TI - Effects of incorporation of hydroxyapatite and fluoroapatite nanobioceramics into conventional glass ionomer cements (GIC). AB - Hydroxyapatite (HA) has excellent biological behavior, and its composition and crystal structure are similar to the apatite in the human dental structure and skeletal system; a number of researchers have attempted to evaluate the effect of the addition of HA powders to restorative dental materials. In this study, nanohydroxy and fluoroapatite were synthesized using an ethanol based sol-gel technique. The synthesized nanoceramic particles were incorporated into commercial glass ionomer powder (Fuji II GC) and were characterized using Fourier transform infrared and Raman spectroscopy, X-ray diffraction and scanning electron microscopy. Compressive, diametral tensile and biaxial flexural strengths of the modified glass ionomer cements were evaluated. The effect of nanohydroxyapatite and fluoroapatite on the bond strength of glass ionomer cement to dentin was also investigated. Results showed that after 1 and 7 days of setting, the nanohydroxyapatite/fluoroapatite added cements exhibited higher compressive strength (177-179MPa), higher diametral tensile strength (19-20MPa) and higher biaxial flexural strength (26-28MPa) as compared with the control group (160MPa in CS, 14MPa in DTS and 18MPa in biaxial flexural strength). The experimental cements also exhibited higher bond strength to dentin after 7 and 30 days of storage in distilled water. It was concluded that glass ionomer cements containing nanobioceramics are promising restorative dental materials with both improved mechanical properties and improved bond strength to dentin. PMID- 17921078 TI - Callus mineralization and maturation are delayed during fracture healing in interleukin-6 knockout mice. AB - IL-6 is a pleiotropic cytokine involved in cell signaling in the musculoskeletal system, but its role in bone healing remains uncertain. The purpose of this study was to examine the role of IL-6 in fracture healing. Eight-week-old male C57BL/6 and IL-6 -/- mice were subjected to transverse, mid-diaphyseal osteotomies on the right femora. Sacrifice time points were 1, 2, 4, or 6 weeks post-fracture (N=14 per group). Callus tissue properties was analyzed by microcomputed tomography (micro-CT) and Fourier transform infrared imaging spectroscopy (FT-IRIS). Cartilage and collagen content, and osteoclast density were measured histologically. In intact unfractured bone, IL-6 -/- mice had reduced crystallinity, mineral/matrix ratio, tissue mineral density (TMD), and bone volume fraction (BVF) compared to wildtype mice. This suggests that there was an underlying deficit in baseline bone quality in IL-6 -/- mice. At 2 weeks post fracture, the callus of IL-6 -/- mice had reduced crystallinity and mineral/matrix ratio. These changes were less evident at 4 weeks. At 2 weeks, the callus of the IL-6 -/- mice had an increased tissue mineral density (TMD), an increased cartilage and collagen content, and reduced osteoclast density compared to these parameters in wildtype mice. By 4 and 6 weeks, these parameters were no longer different between the two strains of mice. In conclusion, IL-6 -/- mice had delayed callus maturity, mineralization, and remodeling compared with the callus of the wildtype mice. These effects were transient indicating that the role of IL-6 appears to be most important in the early stages of fracture healing. PMID- 17921079 TI - Regional variations in mineralization and strain distributions in the cortex of the human mandibular condyle. AB - The strain (i.e. deformation) history influences the degree of mineralization of cortical bone (DMB) as well as its osteonal microstructure. This study aimed to examine the relationships of stress and strain distributions with the variations in DMB and the osteonal orientations in the cortical bone of the human mandibular condyle. It was hypothesized that strains are inversely proportional to local DMB and that the principal strains are oriented parallel to the osteons. To test this, ten human mandibular condyles were scanned in a microCT system. Finite element models were created in order to simulate static clenching. Within each condyle, 18 volumes of interest were selected to analyze regional differences in DMB, stress and strains. Subchondral bone showed a lower equivalent strain (2652+/-612 muepsilon) as compared to the anterior (p=0.030) and posterior cortex (p=0.007) and was less mineralized. Contrary to our hypothesis, the results show that strains correlated positively with regional variations in DMB (r=0.750, p<0.001). In the anterior and the posterior cortex, the first principal strain was parallel to the cortical surface and oriented supero-inferiorly with a fan like shape. In subchondral bone, the first and the second principal strain were parallel to the surface and oriented antero-posteriorly and medio-laterally, respectively. It was concluded that the strain distributions, by themselves, cannot explain the regional differences found in DMB. In agreement with our second hypothesis, the orientation of the osteonal network of the mandibular condyle was closely related to the strain orientations. The results of this study suggest that the subchondral and the cortical bone are structured to ensure an optimal load distribution within the mandibular condyle and have a different mechanical behaviour. Subchondral bone plays a major role in the transmission of the strains to the anterior and posterior cortex, while these ensure an optimal transmission of the strains within the condylar neck and, eventually, to the mandibular ramus. PMID- 17921081 TI - Contrast agents for hepatic MRI. AB - Liver specific contrast media (LSCM) can be subdivided according to different modalities of hepatic distribution: exclusive distribution to the hepatocellular compartment can be obtained using CM which accumulate within the hepatocytes after slow infusion; other CM demonstrate combined perfusion and hepatocyte selective properties, with an initial distribution to the vascular-interstitial compartment (in an analogous manner to that of the conventional extracellular CM), thereafter, a fraction of the injected dose is taken up into the hepatocytes causing an increase in the signal intensity of the hepatic tissue. The use of the superparamagnetic effect of iron oxide particles is based on distribution in the reticuloendothelial system (RES), usually well represented in the normal parenchyma as well as in benign hepatocellular lesions, and absent in most malignant lesions. It is necessary to have an in-depth knowledge of either the biological and histological characteristics of focal liver lesions (FLL) or the enhancement mechanism of LSCM to gain significant accuracy in the differential diagnosis of FLL. Dynamic contrast-enhanced MRI is an important tool in the identification and characterization of FLL. With LSCM it is possible to differentiate benign from malignant lesions and hepatocellular lesions from non hepatocellular lesions with high accuracy. To understand the contrast behaviour after injection of LSCM it is necessary to correlate the contrast enhancement with both the biological and histological findings of FLL. PMID- 17921082 TI - Tumour volume measurement in head and neck cancer. AB - Tumour volume is a significant prognostic factor in the treatment of malignant head and neck tumours. Studies of laryngeal and pharyngeal tumours have shown tumour volume to be an important predictor for tumour recurrence. Some studies (for instance nasopharyngeal carcinoma) have shown through multivariate modelling that tumour volume is a dominant covariate that overwhelms T stage, N stage and stage group. The results of these studies have prompted several investigators to suggest the inclusion of tumour volume as an additional prognostic factor in future revisions of the TNM staging system. This paper briefly reviews the TNM system as a staging tool, the measurement of tumour volume and how tumour volume could possibly be incorporated in the system or used as an additional prognostic factor. PMID- 17921080 TI - Distinguishing benign from malignant liver tumours. AB - Liver masses are very common and most are benign. It is therefore important to avoid unnecessary interventions for benign lesions, while at the same time ensuring accurate diagnosis of hepatic malignancies. Many cancer patients, like the general population, have incidental benign liver lesions. In planning treatment for cancer patients, it is critical to avoid inappropriate treatment decisions based on misdiagnosis of a benign lesion as a metastasis or primary liver malignancy. This article describes the salient imaging features of the common benign liver masses and outlines a general approach to distinguishing between benign and malignant hepatic lesions. PMID- 17921083 TI - Staging of oesophageal cancer. AB - Diagnosis of oesophageal cancer is usually made at endoscopic biopsy. Imaging is used to stage the tumour, assess response to therapy, and detect complications of therapy and recurrence. For therapy planning, differentiation of resectable (T1 T3, N0, localised N1) versus irresectable disease (T4, extensive N1, M1) is important. Endoscopic ultrasound (EUS) is the method of choice for diagnosing T1 T3 stages, and N0 versus N1, including endoscopic ultrasound (EUS)-guided fine needle aspiration. Computed tomography (CT) or magnetic resonance imaging (MRI) are used to demonstrate infiltration of adjacent structures, distant lymphadenopathy and distant metastases, however, positron emission tomography (PET) and PET-CT are superior in this respect. If imaging suggests irresectable disease, histologic confirmation may be required in order not to prevent curative resection in false positive findings. PMID- 17921084 TI - Whole body MRI and PET/CT in haematological malignancies. AB - The usefulness of whole body magnetic resonance imaging (MRI) and positron emission tomography (PET)/computed tomography (CT) in haematological malignancies is reviewed. PET/CT combining functional and anatomical information is currently a valuable tool in the management of patients with lymphoma, especially in the assessment of early treatment response. MRI is advantageous in evaluating bone marrow involvement and therefore plays an important role in clinical decision making for patients with myeloma. The development of whole body functional MR studies is underway and can potentially complement the PET/CT for better patient care. PMID- 17921086 TI - Defining the target for radiotherapy of head and neck cancer. AB - The greatest challenge for radiation therapy is to attain the highest probability of cure with the least morbidity. Implementation of conformal radiotherapy techniques offers the possibility to target irregularly shaped volumes while optimally sparing the normal tissues. This implies however an accurate knowledge of the exact tumour extension. In order to perfectly delineate the primary tumour and to optimise the radiation dose administered to normal tissues, it is necessary for patients to undergo imaging studies. Both anatomical and functional imaging studies are currently being evaluated for treatment planning of head and neck cancer. PMID- 17921087 TI - Pancreatic adenocarcinoma: diagnosis and staging using multidetector-row computed tomography (MDCT) and magnetic resonance imaging (MRI). AB - Pancreatic adenocarcinoma continues to be a leading cause of cancer death in the Western world and is amongst the leading gastrointestinal cancers. The incidence of pancreatic cancer has been stable or slowly rising in the past few decades. Overall the prognosis is poor with 5-year survival rates still under 5%. Therefore early detection and accurate staging of these tumors is crucial for optimal treatment. PMID- 17921085 TI - PET/CT in oncology: for which tumours is it the reference standard? AB - Positron emission tomography (PET)/computed tomography (CT) has a growing role in the imaging of many cancers. As our experience has grown over the past number of years so has our understanding for which cancers it is particularly useful. The value of PET/CT at each stage of the cancer journey is different for each cancer. This review attempts to tease out the role of PET/CT in the common cancers with particular emphasis on where it is the imaging investigation of choice. PMID- 17921088 TI - Colorectal cancer: imaging surveillance following resection of primary tumour. AB - Most patients with colorectal cancer undergo treatment with curative intent and subsequently enter a surveillance programme. The primary aim of surveillance is to identify patients with disease relapse at a resectable stage. However, the identification of local recurrence and metachronous carcinoma are also important aspects of follow up. Patients under observation may be referred for imaging either because regular imaging forms part of the surveillance strategy, or because tumour relapse is suggested by the development of new symptoms or a rise in tumour markers. This paper reviews the use of new and existing imaging techniques during surveillance following resection of primary colorectal cancer. The use of imaging for this surveillance is an application of cancer imaging that is supported by evidence-based clinical guidelines. Computed tomography provides the mainstay modality on grounds of good overall diagnostic performance combined with high availability and low cost. Improvements in survival with more aggressive follow up and treatment are likely to demand more accurate imaging techniques in the future. PMID- 17921089 TI - PET/CT imaging: what radiologists need to know. AB - Positron emission tomography (PET)/computed tomography (CT) imaging is frequently requested in Oncology. Radiologists and nuclear medicine physicians are often asked to perform a panel of imaging examinations as part of the initial staging or follow-up of cancer patients. Medical imaging must therefore integrate polyvalent skills enabling imaging specialists to understand and interpret all types of images. In this context, PET imaging combined with non-enhanced CT, and diagnostic quality contrast-enhanced CT scan and optimisation of CT settings, is part of this multidisciplinary approach requiring the specific skills of a radiologist and a nuclear medicine physician. This approach must therefore be conducted in both directions: radiologists and nuclear medicine physicians should both know how to correlate PET and CT images, while preserving the specificities of each discipline. Radiologists need to be aware of several aspects of PET imaging: PET technology, the examination procedure and injection of iodinated contrast agent for high quality diagnostic CT, ideally followed by double interpretation of CT images, PET images and fused images. Radiologists should be familiar with PET imaging, as this procedure may be associated with several pitfalls and artefacts that need interpretation by a trained specialist. The authors analyse the examination technique of PET combined with non-enhanced and/or contrast-enhanced CT and the proposals for optimal interpretation of normal or pathological PET/CT fusion images. PMID- 17921090 TI - Multimodality imaging of head and neck cancer. AB - Imaging has an important role in staging, planning treatment and post-treatment follow up of patients with head and neck cancer. The most commonly utilized imaging modalities are discussed in relation to their relative indications, strengths and weaknesses. This is followed by a brief update on the advances in functional magnetic resonance imaging. PMID- 17921091 TI - Imaging of anterior mediastinal masses. AB - Multiple different types of anterior mediastinal masses may be encountered on computed tomography (CT) imaging, and many of these lesions are neoplastic in etiology. These include masses arising from the thymus, thyroid and parathyroid glands, as well as lymph nodes, pericardium, and vessels and nerves. Often, the CT attenuation of the mass can be helpful in narrowing down the differential diagnosis, and attenuation values suggesting fat, water or calcium may suggest certain diagnoses; significant enhancement of the mass with intravenous contrast may also be a helpful feature. Lesions with fatty attenuation include teratomas, thymolipomas and Morgagni hernias. Lesions that may manifest the attenuation of water include pericardial and thymic cysts, abscesses, and lymphangiomas, as well as neurogenic and germ cell tumors. Multiple types of lesions may contain calcium, including thyroid goiters and cancers, thymomas, thymic carcinomas and carcinoids, treated lymphoma, germ cell tumors, parathyroid adenomas, and lymph nodes involved with silicosis, sarcoid, tuberculosis, fungal diseases and pneumocystis. Contrast enhancement may be seen in lesions of vascular origin and in vascular neoplasms, such as parathyroid adenomas and Castleman's disease. In addition to CT attenuation values, the exact location and morphology of the mass in question, in conjunction with clinical features such as patient age, gender, signs, symptoms, and laboratory values, can usually lead to a short list of possible etiologies, thereby directing appropriate additional diagnostic procedures or therapeutic approaches. PMID- 17921092 TI - PET/CT and cross sectional imaging of gynecologic malignancy. AB - Gynecologic cancers are a common cause of morbidity and mortality in women of all ages. While many gynecologic cancers are staged clinically using the International Federation of Gynecology and Obstetrics (FIGO) staging system, imaging can be a useful adjunct to clinical staging. Cross sectional imaging techniques such as ultrasound (US), computed tomography (CT) and magnetic resonance imaging (MRI) have been used to detect and follow patients with gynecologic cancer. These imaging modalities can show anatomic detail and morphologic changes in the female genitourinary tract to good advantage. Positron emission tomography (PET) differs in that it shows functional information that is not easily obtained by the other cross sectional imaging techniques. The fusion of PET with CT allows anatomic localization of functional abnormalities in the female genital tract and thereby allows the detection of gross disease in many malignant conditions both within and outside the confines of the female pelvis. The utility and limitations of imaging common gynecologic tumors such as cervical, ovarian and endometrial cancer are discussed with particular emphasis on PET/CT imaging. PMID- 17921093 TI - Biliary tract neoplasms: diagnosis and staging. AB - Most biliary tract neoplasms are malignant and have been traditionally divided into cancers of the gallbladder, the extrahepatic bile ducts, and ampulla of Vater. Although infrequent, bile duct carcinomas and cancer of the gallbladder are not rare. In the United States, an estimated 6000-7000 new cases of carcinoma of the gallbladder and 3000-4000 new cases of carcinoma of the bile ducts are diagnosed annually. Familiarity with the imaging characteristics of gallbladder and bile duct neoplasms is important to expedite the diagnosis and appropriate treatment of patients who often present with non-specific symptoms of right upper quadrant pain, jaundice, and weight loss. PMID- 17921094 TI - The indeterminate adrenal mass in patients with cancer. AB - With the increasing use of abdominal cross-sectional imaging, incidental adrenal masses are frequently detected. The commonest clinical question is whether these are benign adenomas or malignant primary or secondary masses. The nature of incidentally detected adrenal masses can be determined with a high degree of accuracy using computed tomography (CT) and magnetic resonance imaging (MRI) as benign adrenal masses such as myelolipomas, lipid-rich adenomas, adrenal cysts and adrenal haemorrhage which have pathognomonic imaging findings. However, there remains a significant overlap between the imaging features of some lipid-poor adenomas and malignant lesions. We review the recent advances in CT, MRI and positron emission tomography (PET) which can be used to distinguish between benign adenomas and malignant lesions of the adrenal gland. PMID- 17921095 TI - Musculoskeletal primary tumours: treatment evaluation and detection of recurrences. AB - The role of imaging in treatment evaluation and detection of recurrences of musculoskeletal primary tumours is discussed. PMID- 17921096 TI - Oesophageal cancer: assessment of response and follow up. AB - The prognosis for oesophageal cancer is poor with a median survival of 3-5 months and recurrences are frequent. The best chance of cure is successful surgery and pre-operative chemoradiotherapy is used to try and improve outcomes. However, patients may either not respond or may progress during therapy and it is important to differentiate the responders from non-responders. Clinical parameters such as weight gain and improvement in swallowing can be assessed but imaging is used in an attempt to improve outcomes. PMID- 17921098 TI - Post-treatment imaging of liver tumours. AB - In the past few years, great improvements have been made to achieve local tumour control of primary liver malignancies and liver metastases. For hepatocellular carcinoma (HCC), transarterial chemoembolisation (TACE) and tumour ablation techniques, including percutaneous ethanol injection (PEI), radiofrequency ablation (RF), and laser-induced interstitial thermotherapy (LITT) have been developed. For colorectal liver metastases, surgery is still the standard technique in localised disease, although percutaneous RF ablation has gained considerable acceptance. In patients with widespread disease, chemotherapy with new drugs offers improved survival. Contrast-enhanced computed tomography (CT) and magnetic resonance imaging (MRI) are the modalities of choice to evaluate treatment response. The present review demonstrates imaging findings of complete and incomplete tumour control after intervention as well as the imaging spectrum of complications. Imaging guidelines according to the World Health Organization and Response Evaluation Criteria In Solid Tumors (RECIST) for assessment of chemotherapy response are presented. PMID- 17921099 TI - An update into the pathophysiological role of HER2 in cancer: therapeutic implications. PMID- 17921100 TI - The arsenal of biological therapies. PMID- 17921097 TI - MR imaging in ovarian cancer. AB - Magnetic resonance (MR) imaging is increasingly being used in patients with gynaecological disorders due to its high contrast resolution compared to computed tomography (CT) and ultrasound. In women presenting with an adnexal mass, ultrasound remains the primary imaging modality in the detection and characterisation of such lesions. However, in recent years overwhelming evidence has accumulated for the use of MR imaging in patients with indeterminate adnexal masses particularly in younger women and where disease markers are unhelpful. In staging ovarian cancer and for evaluating therapeutic response MR imaging is as accurate as CT but CT remains the imaging modality of choice because it is more widely available and quicker. This article reviews that evidence and outlines a place for the use of MR imaging in ovarian cancer. PMID- 17921101 TI - Molecular markers in colorectal cancer: genetic bases for a customised treatment. AB - Colorectal cancer (CRC) is the second leading cause of cancer death in Western countries. CRC treatment is based on the employment of three chemotherapeutic drugs, including 5-fluorouracil, oxaliplatin and irinotecan, and the use of recently incorporated targeted agents directed to vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR). The approval of these biologicals and of others to come holds great promise for the improvement of patient outcome. The molecular bases for this lethal disease have been extensively investigated, laying the foundations for a rational and customised treatment approach, expanding the therapeutic index of current drugs and easing the incorporation of new molecules. Individual markers have been mainly investigated based on drug targets and metabolism. Also, the increasing availability of highthroughput technologies has prompted the opportunity for blind studies capable of screening new markers and of identifying the specific oncogenic pathways responsible for drug resistance in a given patient. An updated review of the field is presented in this article. PMID- 17921104 TI - Effect of surgical resection of metastatic disease on immune tolerance to cancer. How a systemic disease could be controlled by a local therapy. AB - Surgery represents the only chance of cure for patients with colorectal liver metastases. The results of expanded indications for surgical treatment revealed that even advanced disease can be cured in a significant percentage of cases. What is the explanation for this systemic impact of a local treatment such as surgery? What is different in those patients who can be cured by resection? In this review we analyse the available evidence of the complex relationship between the growing tumour and the immune system. Special attention is directed to the role of T regulatory cells (Tregs) recruited by the tumour to construct a tolerogenic microenvironment in which to grow. Based on the published data we developed the hypothesis that surgery breaks the tumour immune tolerance status because it not only removes the tumour, but also the protective shield of T regulatory cells. PMID- 17921102 TI - Apoptosis regulators as targets for cancer therapy. AB - Apoptosis serves to remove excess or damaged cells and its dysregulation may lead to a number of pathological disorders including cancer. Studies during the last 20 years have unravelled much of the molecular mechanisms that control apoptosis. Whether a cell dies in response to diverse apoptotic stimuli, including DNA damaging agents, is determined largely by interactions between proteins of the Bcl-2 family. A death signal is transmitted through the BH3-only proteins to Bax and Bak which in turn permeabilise the outer mitochondrial membrane allowing the release of apoptogenic factors, which triggers activation of cell-deathpromoting caspases. These proteolytic enzymes are tightly controlled by members of the inhibitor of apoptosis (IAP) family. Activation of the caspase cascade via cell death receptors also represents a key apoptotic pathway in both normal and tumour cells. Basic knowledge of these apoptosis regulators provides the basis for novel therapeutic strategies aimed at promoting tumour cell death or enhancing susceptibility to apoptotic inducers. This review focuses on these strategies. PMID- 17921105 TI - Docetaxel-induced interstitial pneumonitis following non-small-cell lung cancer treatment. AB - Interstitial pneumonitis has been described infrequently following administration of docetaxel, used alone or in combination with other chemotherapeutic agents or concurrent irradiation, for non-small-cell lung cancer (NSCLC). This toxicity is of special relevance in NSCLC, as clinical severity and differential diagnosis may be especially challenging. It seems to be due to type I and type IV hypersensitivity reactions to the drug. Clinical and radiographic features are nonspecific and diagnosis is made by exclusion. The rate of grade III-IV docetaxel-induced pneumonitis, ranging from 7 to 47%, depends on several factors, including total dose, chemotherapy schedule and especially concomitant docetaxel treatment with gemcitabine and radiotherapy. Although the usual outcome is cure, it sometimes eventually progresses to pulmonary fibrosis despite steroid treatment. This toxicity must be taken into account when planning treatment strategies for NSCLC in order to reduce its rate and to achieve prompt diagnosis and treatment. PMID- 17921103 TI - Peptidomimetics in cancer chemotherapy. AB - A summary of the current status of the application of peptidomimetics in cancer therapeutics as an alternative to peptide drugs is provided. Only compounds that are used in therapy or at least under clinical trials are discussed, using inhibitors of farnesyltransferase, proteasome and matrix metalloproteinases as examples. PMID- 17921106 TI - Recommendation of the scientific societies on the treatment of anaemia in cancer patients. AB - At present, anaemia in the patients with cancer remains a problem of the first magnitude and of increasing interest due to the high incidence, the major knowledge of its physiopathology, the negative impact on the quality of life of the patient, the influence on the evolution of the disease and its treatments and, finally to the progressive development of new alternatives of treatment, especially the erythropoietic agents. For all this, it becomes necessary to consider the treatment of the anemia of the patients with cancer as a basic part of their support treatment. The erythropoietic agents have demonstrated in the last years that constitute a therapeutic alternative to obtain an increase of the levels of hemoglobin in the patients with anticancer treatments, considering specially that the correction of the anemia not only represents the improvement of an analytical value but also has a significant impact on the quality of life of the patients and diminishes the transfusion requirements. Erythropoietic proteins available for the treatment of the anemia of the patients with cancer are Epoetin-alpha, Epoetin-beta and Darbepoetin-alpha. The existence of different drugs, different doses and intervals of administration, clinical different situations and heterogeneous studies, made necessary the development of documents of consensus and guides of clinical recommendations which provide information on the scientific evidence that supports the use of these agents in medical care. This paper summarizes the main recommendations from panels of experts and scientific societies published so far. PMID- 17921107 TI - Incidence of radiation-induced leukoencephalopathy after whole brain radiotherapy in patients with brain metastases. AB - INTRODUCTION: Whole brain radiation therapy (WBRT) remains a recommended treatment for patients with brain metastases in terms of symptom palliation, especially when extracranial systemic disease is present. The aim of the study was to determine the clinical correlation between pre-existing leukoaraiosis and posterior leukoencephalopathy secondary to WBRT. METHODS AND MATERIALS: We retrospectively reviewed the results of WBRT treatment in 44 patients with melanoma brain metastases. The neuroimaging abnormalities of the white matter (T2 weighted MRI) were graded over time. RESULTS: From the 37 evaluable patients the mean age was 53 years old, 23 male and 14 female. Vascular risk factors were present in 22 patients (59.5%). The WBRT total dose was 20 Gy/5fr (n=21) and 30 Gy/10fr (n=16). Leukoaraiosis pre-WBRT was observed in 9/37 patients (24.3%) and leukoencephalopathy post-WBRT in 2/37 (5.4%). Univariate analysis of prognostic factors (sex, age and vascular risk factors) for leukoaraiosis was conducted observing statistically significant differences for patients with age>or=65 years old (p=0.003). Nineteen patients survived more than 3 months. Twelve patients (63.2%) suffered from vascular risk factors. Univariate analysis demonstrated previous leukoaraiosis as a prognostic factor for developing further leukoencephalopathy after WBRT (p=0.015). CONCLUSIONS: Radiation-induced leukoencephalopathy is greater in patients with pre-existing leukoaraiosis. Because of the potential of long-term survival in a small subset of patients with brain metastases and the risk of radiation-induced dementia, neurotoxicity reduction in patients with leukoaraiosis is an important goal of treatment. PMID- 17921109 TI - Pancreatic metastases from ductal and lobular carcinomas of the breast. AB - The pancreas is an uncommon target for metastases from other primary tumours. We discuss clinical, diagnostic and therapeutic aspects of pancreatic metastases through presentation of two surgically treated cases of metastases originating from breast cancer. PMID- 17921110 TI - Perianal metastases from lobular breast carcinoma. AB - Breast cancer gastrointestinal and soft tissue metastases are extremely rare. We present the case of a woman with perianal metastases from a primary lobular breast carcinoma 11 years after mastectomy and local radiotherapy. PMID- 17921108 TI - Predictive factors for radiation-induced pulmonary toxicity after three dimensional conformal chemoradiation in locally advanced non-small-cell lung cancer. AB - BACKGROUND AND PURPOSE: Radiation pneumonitis (RP) is a restricting complication of non-small-cell lung cancer irradiation. Three-dimensional conformal radiotherapy (3D-CRT) represents an advance because exposure of normal tissues is minimised. This study tries to identify prognostic factors associated with severe RP. MATERIALS AND METHODS: Eighty patients with stage IIIA (20%) and IIIB (80%) NSCLC treated with cisplatin- based induction chemotherapy followed by concurrent chemotherapy and hyperfractionated 3D-CRT (median dose: 72.4 Gy, range: 54.1 85.9) were retrospectively evaluated. Acute and late RP were scored using RTOG glossary. Potential predictive factors evaluated included clinical, therapeutic and dosimetric factors. The lungs were defined as a whole organ. Univariate and multivariate analyses were performed. RESULTS: Early and late RP grade>or=3 were observed in two patients (2%) and 10 patients (12%), respectively. Five patients (6%) died of pulmonary toxicity, 3 of whom had pre-existing chronic obstructive pulmonary disease (COPD). Median time to occurrence of late RP was 4.5 months (range: 3-8). Multivariate analysis showed that COPD (OR=10.1, p=0.01) and NTCPkwa>30% (OR=10.5, p=0.007) were independently associated with late grade>or=3 RP. Incidence of RP>or=3 grade for patients with COPD and/or NTCPkwa>30% was 25% vs. 4% for patients without COPD and NTCPkwa<30% (p=0.01). Risk of severe RP was higher for patients with COPD and/or NTCPkwa>30% (OR=7.3; CI 95%=1.4-37.3, p=0.016). CONCLUSIONS: COPD and NTCP are predictive of severe RP. Careful medical evaluation and meticulous treatment planning are of paramount importance to decrease the incidence of severe RP. PMID- 17921111 TI - Phagosome maturation in unicellular eukaryote Paramecium: the presence of RILP, Rab7 and LAMP-2 homologues. AB - Phagosome maturation is a complex process enabling degradation of internalised particles. Our data obtained at the gene, protein and cellular level indicate that the set of components involved in this process and known up to now in mammalian cells is functioning in unicellular eukaryote. Rab7-interacting partners: homologues of its effector RILP (Rab-interacting lysosomal protein) and LAMP-2 (lysosomal membrane protein 2) as well as alpha7 subunit of the 26S proteasome were revealed in Paramecium phagolysosomal compartment. We identified the gene/transcript fragments encoding RILP-related proteins (RILP1 and RILP2) in Paramecium by PCR/RT-PCR and sequencing. The deduced amino acid sequences of RILP1 and RILP2 show 60.5% and 58.3% similarity, respectively, to the region involved in regulating of lysosomal morphology and dynein-dynactin recruitment of human RILP. RILP colocalised with Rab7 in Paramecium lysosomes and at phagolysosomal membrane during phagocytosis of both the latex beads and bacteria. In the same compartment LAMP-2 was present and its expression during latex internalisation was 2.5-fold higher than in the control when P2 protein fractions (100,000 x g) of equal load were quantified by immunoblotting. LAMP-2 cross reacting polypeptide of approximately106 kDa was glycosylated as shown by fluorescent and Western analysis of the same blot preceded by PNGase F treatment. The alpha7 subunit of 26S proteasome was detected close to the phagosomal membrane in the small vesicles, in some of which it colocalised with Rab7. Immunoblotting confirmed presence of RILP-related polypeptide and a7 subunit of 26S proteasome in Paramecium protein fractions. These results suggest that Rab7, RILP and LAMP-2 may be involved in phagosome maturation in Paramecium. PMID- 17921112 TI - Relationship between growth hormone concentrations in bovine oocytes and follicular fluid and oocyte developmental competence. AB - In the last few years, several works suggest that Growth Hormone (GH) is involved in follicular development and oocyte maturation. These actions may reflect endocrine roles of pituitary GH and also account for local autocrine or paracrine activities of GH produced in reproductive tissue. This study was aimed to verify whether the developmental competence of bovine female gametes might be related to ovarian GH. We evaluated the localisation and distribution of GH in the cumulus oocytes complexes (COCs) and the concentration of GH in the oocytes and in the follicular fluids (FF) from ovaries classified on the basis of the follicles number. Oocytes retrieved from ovaries with more than 10 follicles of 2 to 5 mm in diameter (High ovaries, Hi) show higher rate of maturation and blastocyst formation than those retrieved from ovaries with less than 10 follicles (Low ovaries, Lo). At the same time we measured Estrogen (E2) and Progesterone (P4) concentrations in FF, to relate oocytes quality, GH concentration and follicle health. GH localization in COCs and oocytes was performed by indirect immunofluorescence and its concentration within the ooplasm was evaluated by microspectrophotometer analysis. GH, E2 and P4 concentrations in FF were measured by an Enzyme Linked ImmunoSorbent assay (ELISA). We observed a positive, diffuse signal at cytoplasmic level in most of the cumulus cells, with no differences between COCs collected from Hi and Lo ovaries. On the contrary, GH level was significantly higher in the oocytes collected from Lo ovaries than in those recovered from Hi ovaries. Finally we found that also GH level in the FF was inversely related to the oocytes developmental capability. We suggest that the increase of GH in the oocytes and in the FF derived from Lo ovaries might be interpreted as attempt of the follicular environment to improve ovarian activity and in turn oocytes developmental competence in a autocrine-paracrine manner. Moreover, E2, and P4 levels in FF suggest that, in our model, atresia processes are also involved in oocyte developmental capability and that the highest level of GH may represent a local reaction to these phenomena. PMID- 17921113 TI - N-cadherin is essential for retinoic acid-mediated cardiomyogenic differentiation in mouse embryonic stem cells. AB - Contraction forces developed by cardiomyocytes are transmitted across the plasma membrane through end-to-end connections between the myocytes, called intercalated disks, which enable the coordinated contraction of heart muscle. A component of the intercalated disk, the adherens junction, consists of the cell adhesion molecule, N-cadherin. Embryos lacking N-cadherin die at mid-gestation from cardiovascular abnormalities. We have evaluated the role of N-cadherin in cardiomyogenesis using N-cadherin-null mouse embryonic stem (ES) cells grown as embryoid bodies (EBs) in vitro. Myofibrillogenesis, the spatial orientation of myofibers, and intercellular contacts including desmosomes were normal in N cadherin-null ES cell-derived cardiomyocytes. The effect of retinoic acid (RA), a stage and dose-dependent cardiogenic factor, was assessed in differentiating ES cells. all-trans (at) RA increased the number of ES cell-derived cardiomyocytes by approximately 3-fold (at 3 x 10(-9) M) in wt EBs. However, this effect was lost in N-cadherin-null EBs. In the presence of supplemented at-RA, the emergence of spontaneously beating cardiomyocytes appeared to be delayed and slightly less efficient in N-cadherin-null compared with wt and heterozygous EBs (frequencies of EBs with beating activity at 5 days: 54+/-18% vs. 96+/-0.5%, and 93+/-7%, respectively; peak frequencies of EBs with beating activity: 83+/-8% vs. 96+/ 0.5% and 100%, respectively). In conclusion, cardiomyoyctes differentiating from N-cadherin-null ES cells in vitro show normal myofibrillogenesis and intercellular contacts, but impaired responses to early cardiogenic effects mediated by at-RA. These results suggest that N-cadherin may be essential for RA induced cardiomyogenesis in mouse ES cells in vitro. PMID- 17921114 TI - Expression of hepatocyte growth factor in Hashimoto's thyroiditis with nodular lesions. AB - Hashimoto's thyroiditis (HT) is an autoimmune thyroid disease frequently associated with hyperplastic nodules (HN)s. Hepatocyte growth factor (HGF) is expressed in benign thyroid nodules and over-expressed in malignant thyroid nodules, particularly in papillary thyroid carcinomas. To elucidate the role of HGF in the development of HNs in association with HT we evaluated, by immunohistochemistry, the expression of HGF in both nodular and extranodular tissues, obtained from 30 HTs and 15 goiter samples. Six normal thyroid glands were used as controls. All normal control tissue samples exhibited no evidence of HGF immunoreaction. HNs showed weak to moderate HGF immunoreaction, which was located exclusively in the cytoplasm of stromal cells (fibroblasts and endothelial cells). However, the percentage of positive cases was higher in HNs arisen in the context of HT, compared to HNs not associated with HT (30/30 or 100% vs 4/15 or 40%; p<0.001). HGF immunoreactivity was also detected in all extranodular tissues from HT specimens (30/30 or 100%), but we found some significant differences. In fact, while in HNs observed in the context of HT lesions HGF was expressed only in stromal cells, in the extranodular tissues from the same thyroid gland affected by HT it was also detected in the cytoplasm of the epithelial follicular cells. Furthermore, HTs showed a much higher HGF staining grade in the extranodular tissue compared to HNs. Finally, a clear positive correlation was observed in HT between the proportion of HGF expressing follicular cells and the grade of lymphoid aggregates of the thyroid gland. In conclusion, HGF is much more frequently and highly expressed in thyroid tissue with HT, compared to goiter. In HT glands HGF can be detected in both follicular thyroid cells and stromal cells, while in HNs, either from goiters or associated with HT, its expression is restricted only to the stromal cells. These data indicate that HGF may play a role in cell proliferation processes occurring in thyroid glands affected by HT, probably under the regulation of the lymphoid infiltrate. PMID- 17921115 TI - The expression of innate immunity genes in Italian Crohn disease patients. AB - Crohn' disease (CD) is a chronic idiopathic inflammatory bowel disease characterized by the interaction of both hereditary and environmental factors. Intestinal flora and pathogens such as bacteria, viruses and fungi, are thought to be the first step leading to an inflammatory status, which is subsequently amplified in genetically susceptible patients thus triggering the disease. Since the innate immune system is believed to be very important in regulating the flora of the gastrointestinal tract, we decided to study the influence of two important molecules of the innate immune system in CD. Frozen intestinal biopsies from 49 Crohn patients and 10 healthy individuals were collected at the gastroenterology unit of Children's Hospital Burlo Garofolo in Trieste and innate immunity gene expression was evaluated by using both in situ RT-PCR and quantitative PCR. We have analyzed the expression and localization of both MBL2 and DEFB1 genes in intestinal biopsies of Italian Crohn patients by in situ RT-PCR and quantitative PCR. DEFB1 is expressed equally in all subjects. Importantly, MBL2 transcripts were upregulated in CD patients compared to healthy controls. MBL2 expression in controls is normally extremely low, detectable only by quantitative PCR with a Taqman probe. We demonstrated the MBL2 and DEFB1 expression in intestinal biopsies of patients suffering from CD. Our results showed that the MBL2 gene is expressed by cells in the basal lamina, whilst DEFB1 is expressed by epithelial cells. PMID- 17921116 TI - In vitro exposure of human chondrocytes to pulsed electromagnetic fields. AB - The effect of pulsed electromagnetic fields (PEMFs) on the proliferation and survival of matrix-induced autologous chondrocyte implantation (MACI)-derived cells was studied to ascertain the healing potential of PEMFs. MACI-derived cells were taken from cartilage biopsies 6 months after surgery and cultured. No dedifferentiation towards the fibro- blastic phenotype occurred, indicating the success of the surgical implantation. The MACI-derived cultured chondrocytes were exposed to 12 h/day (short term) or 4 h/day (long term) PEMFs exposure (magnetic field intensity, 2 mT; frequency, 75 Hz) and proliferation rate determined by flow cytometric analysis. The PEMFs exposure elicited a significant increase of cell number in the SG2M cell cycle phase. Moreover, cells isolated from MACI scaffolds showed the presence of collagen type II, a typical marker of chondrocyte functionality. The results show that MACI membranes represent an optimal bioengineering device to support chondrocyte growth and proliferation in surgical implants. The surgical implant of MACI combined with physiotherapy is suggested as a promising approach for a faster and safer treatment of cartilage traumatic lesions. PMID- 17921117 TI - Localization of MTT formazan in lipid droplets. An alternative hypothesis about the nature of formazan granules and aggregates. AB - MTT (3-(4, 5-dimethyl-2-thiazolyl)-2, 5-dihphenyltetrazolium bromide) assay is a widely used method to assess cell viability and proliferation. MTT is readily taken up by cells and enzymatically reduced to formazan, a dark compound which accumulates in cytoplasmic granules. Formazan is later eliminated by the cell by a mechanisms often indicated as exocytosis, that produces characteristic needle like aggregates on the cell surface. The shape of formazan aggregates and the rate of exocytosis change in the presence of bioactive amyloid beta peptides (Abeta) and cholesterol. Though the cellular mechanisms involved in MTT reduction have been extensively investigated, the exact nature of formazan granules and the process of exocytosis are still obscure. Using Nile Red, which stains differentially neutral and polar lipids, and a fluorescent analog of cholesterol (NBD-cholesterol), we found that formazan localized in lipid droplets, consistent with the lipophilic nature of formazan. However, formazan granules and aggregates were also found to form after killing cells with paraformaldehyde fixation. Moreover, formazan aggregates were also obtained in cell-free media, using ascorbic acid to reduce MTT. The density and shape of formazan aggregates obtained in cell-free media was sensitive to cholesterol and Abeta. In cells, electron microscopy failed to detect the presence of secretory vesicles, but revealed unusual fibers of 50 nm of diameter extending throughout the cytoplasm. Taken together, these findings suggest that formazan efflux is driven by physico chemical interactions at molecular level without involving higher cytological mechanisms. PMID- 17921118 TI - Immunolocalization of BRCA1 protein in tumor breast tissue: prescreening of BRCA1 mutation in Tunisian patients with hereditary breast cancer? AB - BRCA1 is a tumor suppressor gene which is inactivated by mutation in familial breast and ovarian cancers. Over 300 different disease causing germ-line mutations have been described; 60% are unique to an individual family. This diversity and the large size of the gene lead us to search for a prescreening method for BRCA1 mutations. Since BRCA1 is a nuclear protein in normal cells, but reported by some authors to be cytoplasmic in breast tumor cells of patients with BRCA1 mutation, we evaluated immunohistochemistry as a prescreening technique to identify BRCA1 mutations in patients with familial presentation of breast cancer. Using a monoclonal antibody against the carboxy-terminal region of BRCA1, we performed immunohistochemistry on 18 tumor samples from patients with hereditary breast cancer. Cytoplasmic staining of BRCA1 was observed in 10 cases. Of the 18 tumors, 12 (66%) showed either BRCA mutation or BRCA1 accumulation or both, indicating that BRCA1 function might be lost in breast tumor cells not only through mutation, but also via abnormal cytoplasmic location. The immunohistochemical test used in this study would not be efficient as a pre screening method of deleterious mutations, but it appeared useful to investigate tumor physiology. PMID- 17921119 TI - [The role of continuous glucose monitoring in the therapy of diabetes mellitus]. AB - INTRODUCTION: Continuous glucose monitoring system is a wide spreading method in the control of the therapy of diabetes, nowadays recording the fluctuation of the glucose level between two measurements during the classical glucose measurements. PATIENTS AND METHOD: 79 measurements of 53 diabetic patients were analysed. 48/53 (90.5%) (30 women, 18 men, mean age: 26.3 +/- 16.8 years) were patients with type 1 diabetes, 5/53 (9.5%) (4 women, 1 man, mean age: 58.4 +/- 6.2 years) were patients with type 2 diabetes. After delineating the technical details of this technique, results of the measurements recorded in the authors' patients population are presented reviewing the advantages and disadvantages of the method. During the average measurements of 3-4 days MiniMed apparatus were used. RESULTS: The method proved to be useful in detecting the down-phenomenon and the asymptomatic hypoglycemic events mainly, however, long hyperglycemic periods were seen several times. The method led to change of therapy in 64.5% of all measurements, but the authors present unsuccessful measurements due to the error of the sensor and cases when the results did not help the therapeutic decision. CONCLUSIONS: Authors describe a wide range of indications of the usage of this method based on their own and other authors' experience presenting its applicability and limits. It is highlighted that the method is not effective in patients with type 2 diabetes. Presenting their results the authors emphasize that this method provides extra information compared to the classical measurements leading to a better glycemic control and life expectancy in type 1 diabetes patients. PMID- 17921120 TI - [Neurological and psychiatric aspects of some endocrine diseases. The role of neurosteroids and neuroactive steroids]. AB - Regardless of their origin, neuroactive steroids are capable of modifying neural activities by modulating different types of membrane receptors. Neurosteroids are synthesized de novo in neurones and glia. Steroidogenic enzymes are found in the central nervous system. Classical steroid receptors are localized in the cytoplasm, they exert regulatory actions on the genome, and their activation causes medium- and long-term effects. Non-classical receptors are located within the membrane and act as mediators of short-term effects. Other important players are co-repressors and co-activators that can interfere with or enhance the activity of steroid receptors. Beyond their function in stress, corticosteroids play a very important role in fear, anxiety, and memory functions. Patients with Cushing's syndrome frequently develop mood disorder, reversible brain atrophy with transient memory loss, rarely delirium or psychosis. Well-known peripheral symptom is steroidal myopathy. In patients with Addison's disease the main signs are weakness of muscles, lack of energy, decreased mental functions and reduced quality of life. Estrogen and progesterone have their own respective hormone receptors, whereas allopregnanolone acts via the GABA receptors. These hormones have significant role in the development of brain, the architecture of neural circuits and dendrites, density of axonal connections, and the number of neurons. They influence maturation, neuroprotection, seizures, cognitive functions, mood, anxiety, pain, and restitution of peripheral nerves. Androgens also affect cognitive functions, pain, anxiety, mood, and additionally aggression. PMID- 17921121 TI - [The role of Orvosi Hetilap in the development of Hungarian gastroenterology. Part 1: 1857-1904]. AB - The role of the Hungarian weekly medical journal Orvosi Hetilap - first published in 1857 - in the development of special medical fields has not been studied. AIM: Analysis of the papers dealing with diseases of the gastrointestinal tract published in Orvosi Hetilap. METHOD: The author manually reviewed the journal volumes between 1857 and 1904. The original articles, journal and book reviews were identified and classified according to their subject and origin. RESULTS: In the period studied, 455 original articles were published about gastrointestinal diseases, constituting 8.2% of the total publications. Diseases of the oesophagus were studied in 2.4% of the articles, gastric disturbances in 14.7%, intestinal diseases in 14.2%, and hepatobiliary diseases in 24.2%. 592 journal articles were reviewed from 113 different publications, 52.5% being German, 15.9% French and 10.0% English; 42.7% of the articles were published in 10 core journals (8 German and 2 British). Many of the 116 books presented became classics of the medical literature. Basic knowledge in endoscopy was mediated by French and German authors. After pioneering laryngoscopic and urologic examinations, the first oesophagoscopies with rigid instruments were performed in 1902. The most studied field was hepatology: the clinical and pathologic features of acute liver failure, chronic hepatitis and cirrhosis were then described. CONCLUSIONS: Most of the contemporary aspects of gastrointestinal diseases were covered by the original articles published in Orvosi Hetilap . German and French and, to a lesser extent, British sources of scientific information prevailed. Diseases of the liver and biliary tract were studied in most detail. PMID- 17921122 TI - [Maternal age as a risk factor in triplet pregnancy]. AB - Upon the diagnosis of triplet conception, pregnant women should be counseled about the risk of triplet pregnancy and the chances of survival of triplet newborns. This information can help women with triplet gestations to make well informed decisions whether to continue their pregnancy or to undergo multifetal pregnancy reduction. AIM: To assess the maternal age as a risk factor in a large representative sample of the Hungarian triplet population with evaluation of maternal complications and perinatal results. METHODS: In a retrospective cohort study we analyzed 114 triplet pregnancies which ended in delivery from July 1st, 1990 until June 30th, 2006. RESULTS: In the study population there were 15 patients aged 35 years or more at delivery (13.2%) and 99 women were younger than 35 years (86.8%). The rate of maternal complications was similar in the two groups, and there was no significant difference in gestational age at delivery (33.0 +/- 2.9 vs. 32.2 +/- 3.3 weeks), very early (< 28 weeks) premature deliveries (6.7% vs. 9.1%) and early (< 32 weeks) premature deliveries (32.9% vs. 43.4%), respectively, although all of these variables were more favorable over 35 years of age. The mean birth weight of live born infants (1796 +/- 492 vs. 1664 +/- 506 g, p = 0.064 g) was higher, the prevalence of very-very-low-birth-weight (< 1000 g) infants (6.8% vs. 10.7%) and very-low-birth-weight (< 1500 g) infants (34.1% vs. 38.6%) was lower in the groups of older patients. The 1- and 5-minute Apgar scores of newborns in the > 35-year group were significantly larger (8.4 +/ 0.5 vs. 8.0 +/- 1.0, p = 0.016 and 9.5 +/- 0.7 vs. 9.2 +/- 0.8, p = 0.006, respectively). There was no difference in perinatal results or puerperal complications between the groups. The incidence of sepsis/pneumonia and need for ventilation/oxygen therapy was lower among the infants of the patients over 35 years (6.9% vs. 28.6%, p = 0,011 and 31.0% vs. 58.2%, p = 0,011, respectively). The rate of other neonatal complications was similar in the two groups. CONCLUSIONS: Although the incidence of some maternal complications during pregnancy and in the puerperal period is higher at older age, the favorable perinatal results and neonatal morbidity rates make older maternal age a risk reducing (protective) factor in triplet pregnancies. PMID- 17921123 TI - [Metabolic bone disease in premature infants and genetic polymorphisms]. AB - Metabolic bone disease is an important complication among infants very-low-birth weight (< 1500 g). In adults, osteoporosis has been shown to be associated with polymorphisms of vitamin D receptor, estrogen receptor, and collagen Ialpha1 receptor genes. AIM: The primary goal of the study was to investigate the possible association between metabolic bone disease and the allelic polymorphisms of these three genes. METHOD: 104 infants very-low-birth-weight were enrolled to the study. Bone formation (serum alkaline phosphatase, osteocalcin) and bone resorption (urinary excretion of calcium and pyridinium crosslink) markers were determined and x-rays of the chest and wrist (together with the distal portions of associated long bones) were obtained. RESULTS: Thirty infants (28,8%) were diagnosed with metabolic bone disease based on high activity of bone formation, bone resorption markers, and positive radiologic signs. Statistically significant correlation between thymine-adenine repeat [(TA) n ] allelic variant of estrogen receptor gene and bone disease was observed. Infants with metabolic bone disease more often carried low number of repeats [(TA) n < 19] [odds ratio (OR): 5.82, 95% confidence interval (CI): 2.26-14.98]. Significantly higher number of repeats [(TA)n > 18] was found more frequently in the control group (OR: 0.20, 95% CI: 0.05-0.82). Furthermore significant interaction between vitamin D receptor and collagen Ialpha1 receptor genotypes ( p = 0.023) was observed. In a forward stepwise logistic regression model, bone disorder of preterms correlated with male gender ( p = 0.001), duration of hospitalization ( p = 0.007), homozygous allelic variants of high number of (TA) n repeats ( p = 0.025) and interaction between vitamin D receptor (Tt) and estrogen receptor (homozygous allelic variants of low number of repeats) genotype ( p = 0.037). CONCLUSION: The results suggest that the development of metabolic bone disease in infants very-low-birth weight may be associated with genetic polymorphisms. PMID- 17921126 TI - Endovascular treatment of thoracic aortic pathologies in patients with aortoiliac occlusive disease. AB - Today, with the increasing experience and advances in graft technology, endovascular grafting is applied nearly to all levels of the aorta for the treatment of various aortic pathologies. The major route of the stent graft deployment is from the femoral region through the iliac arteries. Since both aneurysms and arterial occlusive diseases share atherosclerosis as the common ancestor of etiology, some patients may poses both aneurysms and stenosis together. These stenotic changes occurring in the iliac and femoral arteries may complicate the passage of the stent graft system. In this report, we sought to evaluate an alternative novel route of graft system application for endovascular treatment of thoracic aortic aneurysms. PMID- 17921127 TI - Minimally invasive cardiac surgery using a flexible aortic clamp. AB - Minimally invasive surgery for mitral valve disease has been performed using a variety of technologies, some of which are complex, have a steep learning curve, and are expensive. We have adopted a simple cost-effective approach over the last 7 years to perform a variety of minimally invasive procedures with excellent outcomes. There have been no strokes, no mortalities, and no episodes of limb ischemia in our series. No patient has required reoperation. PMID- 17921128 TI - Multidetector computed tomography findings of arrhythmogenic right ventricular dysplasia: a case report. AB - Arrhythmogenic right ventricular dysplasia (ARVD) is a heart muscle disorder characterized pathologically by fatty or fibrofatty replacement and electrical instability of the right ventricular myocardium. This cardiac entity leads to sudden cardiac death, syncope, recurrent ventricular tachycardia, and in some cases, heart failure in a younger population. Contrast angiography, echocardiography, radionuclide angiography, ultrafast computed tomography (CT), and cardiovascular magnetic resonance imaging are techniques used to diagnose functional and morphologic characteristics of the disease. CT is sensitive in detecting intramyocardial fat because of its low attenuation. Recently the advances in multislice CT (MDCT) have improved temporal resolution, which has increased effectiveness in providing morphologic and functional information. We present a case with ARVD evaluated through 16-row MDCT. Fatty infiltration was clearly demonstrated by 16-slice CT; thus, multislice CT may have a significant role in the assessment and follow-up of patients with ARVD. PMID- 17921129 TI - Can leg pain be the sole presenting symptom of infective endocarditis? A case report. AB - Infective endocarditis is a serious septic disease that can be life threatening unless effective therapy is instituted following the correct diagnosis. The complication of septic embolism and mycotic aneurysm in patients with infective endocarditis may increase morbidity and mortality. We present a case of peroneal artery aneurysm with coincident double native heart valve endocarditis in a patient. PMID- 17921130 TI - Anomalous right subclavian artery with ascending aortic aneurysm: a case report. AB - We report a rare case of the ascending aortic aneurysm with an anomalous origin of the right subclavian artery. The right subclavian artery branched from the aorta as the fourth major vessel and ran behind the esophagus. Moreover, the left and right coronary arteries arose ectopically from the posterior and the left aortic sinus, respectively. PMID- 17921131 TI - Genetic polymorphisms contribute to acute kidney injury after coronary artery bypass grafting. AB - BACKGROUND: Acute kidney injury is one of the most serious complications after cardiac surgery. Genetic polymorphisms are reported to be associated with postoperative renal impairment. The aim of this study was to investigate the relationship between selected gene polymorphisms and acute kidney injury after cardiac surgery. METHODS: Two hundred forty-eight elective coronary artery bypass grafting procedure patients were enrolled in the study. Angiotensin-converting enzyme (ACE) II, ID, and DD, apolipoprotein E (APO E), and angiotensin II type 1 receptor (AGTR1) A1166C genotypes were detected by polymerase chain reaction. Plasma levels of ACE were analyzed by enzyme-linked immunosorbent assay. Acute kidney injury after cardiac surgery was graded according to the RIFLE (risk, injury, failure, loss, and end-stage kidney disease) classification. RESULTS: In our study, 21.8% of patients had acute renal impairment after cardiac surgery. Among the 54 patients with acute kidney injury, ACE D allele frequency was 0.620. The plasma levels of ACE were significantly higher in the D allele carriers (P = .018). Three of the 54 patients with acute kidney injury were APO E epsilon 4 allele carriers (P = .002). AGTR1 C allele carriers constituted 46% of all patients with postoperative acute kidney injury. There was no statistically significant difference between A allele homozygotes and C allele carriers with respect to postoperative renal dysfunction (P > .05). CONCLUSIONS: The present findings support the hypothesis that ACE I/D and APO E gene polymorphisms may play a role in the development of acute kidney injury after cardiac surgery. However, AGTR1 does not have a unique association with postoperative renal impairment. PMID- 17921132 TI - A novel approach for off-pump atrial septostomy applications. AB - Atrial septostomy or septectomy are required to enable atrial mixture in various congenital cardiac lesions. The aim of this article was to introduce a technique where atrial septostomy application could be employed off pump with the aid of a new device. To our knowledge this is the first report for this technique in the literature. We report the results of 7 patients for whom we employed our technique successfully using a new combined device as an alternative to traditional methods. The major advantage of our approach was avoiding detrimental effects of cardiopulmonary bypass applications. PMID- 17921133 TI - A survey of assisted reproductive technology births and imprinting disorders. AB - BACKGROUND: Genomic imprinting is an epigenetic process in which allele-specific gene expression is dependent on the parental inheritance. Although only a minority of human genes are imprinted, those that have been identified to date have been preferentially implicated in prenatal growth and neurodevelopment. Mutations or epimutations in imprinted genes or imprinting control centres are associated with imprinting disorders such as Angelman syndrome (AS) and Beckwith Wiedemann syndrome (BWS). Recently, an increased frequency of assisted reproductive technology (ART) conceptions has been reported in children with BWS and AS. However, the risk of imprinting disorders in ART children is unknown. METHODS: We undertook a survey of 2492 children born after ART in the Republic of Ireland and Central England with the aim of detecting cases (both clinically diagnosed and previously unrecognized) of BWS and AS in this cohort. The response rate to an initial questionnaire was 61%, corresponding to data for 1524 children. After evaluation of the questionnaire, 70 children were invited for a detailed clinical assessment, and 47 accepted (response rate of 67%). RESULTS: In this entire cohort, we detected one case of BWS and no cases of AS. We did not find evidence that there exists a significant group of ART children with unrecognized milder forms of AS or BWS. CONCLUSIONS: Although previous studies have suggested an increased relative risk of BWS and AS after ART, our findings suggest that the absolute risk of imprinting disorders in children conceived by ART is small (<1%). Precise risk estimates of risk are difficult to define because of the rarity of the conditions and incomplete response rates to the questionnaire and clinical examination invitations. Hence further investigations are indicated to (i) refine the absolute and relative risks of imprinting disorders after ART and (ii) ensure that changes in ART protocols are not associated with increased frequencies of epigenetic changes and imprinting disorders in children born after ART. PMID- 17921134 TI - Implantation is apparently unaffected by the dopamine agonist Cabergoline when administered to prevent ovarian hyperstimulation syndrome in women undergoing assisted reproduction treatment: a pilot study. AB - BACKGROUND: Ovarian hyperstimulation syndrome (OHSS) is a result of ovarian overexpression of vascular endothelial growth factor (VEGF) and its receptor 2 (VEGFR2). VEGF/VEGFR2 binding disrupts cellular junctions and increases vascular permeability (VP), a characteristic of OHSS, but enhances angiogenesis, which is a fundamental step in implantation. In animals, the dopamine agonist Cabergoline (Cb2) prevents VP without affecting angiogenesis. In humans, Cb2 averts OHSS, but a possible detrimental effect on angiogenesis and implantation has not been explored. A pilot study was designed to analyze whether or not Cb2 administration, as a procedure for preventing OHSS, affects the outcome of assisted reproduction treatment (ART). METHODS: A retrospective study with endpoints of implantation and ongoing/term pregnancy rates. Women (n = 35) at risk of OHSS (20-30 follicles developed and >20 oocytes collected) took a daily oral dose of 0.5 mg Cb2 for 8 days, beginning on the day of hCG. They were matched with controls treated during the same period and who were similar with respect to age, number and quality of the embryos replaced, embryonic stage at transfer and sperm quality. RESULTS: No difference was detected between the groups in fertilization, implantation or pregnancy rates. A total of 14 ongoing (beyond 32 weeks) or full term pregnancies were registered in each group. No major problem was detected during pregnancy or after delivery in any of these babies. CONCLUSIONS: Administration of Cb2 in order to prevent OHSS is safe and does not appear to affect ART outcome. PMID- 17921135 TI - Menstrual-like changes in mice are provoked through the pharmacologic withdrawal of progesterone using mifepristone following induction of decidualization. AB - BACKGROUND: Cyclic shedding of the endometrium is unique to menstruating species, and mouse menstruation models by physiologic progesterone withdrawal have been previously reported. Since progesterone action ablated pharmacologically may provide more insight into the mechanism of action, a mouse menstruation model using mifepristone was established. METHODS: Mifepristone was administered following oil-induced decidualization in ovarectomized mice primed with hormones. Morphology, hormone levels, leukocytes and apoptosis were evaluated over a period of 48 h after treatment. Vaginal smears were used to monitor bleedings. RESULTS: Mifepristone induced menstrual-like changes. Tissue breakdown was drastic by 16 h, and the decidual zone was shed by 24 h while the mice bled. The endometrium regenerated from 24 h onwards and became completely restored by 48 h. These results are consistent with previous reports. However, although progesterone levels remained constant, estradiol levels increased after the treatment. The CD45(+) cells showed two peaks of increase at 16 h (breakdown phase) and 32 h (regeneration phase) (Leukocyte levels also increased in the unstimulated horns, but no breakdown changes occurred there). Moreover, apoptosis drastically increased by 16 h concurrent with tissue destruction. These results differed from those of the physiologic withdrawal models. CONCLUSIONS: The pharmacologic withdrawal of progesterone by mifepristone successfully provoked a menstrual-like process in mice after artificial decidualization. PMID- 17921136 TI - The role of the fibronectin IGD motif in stimulating fibroblast migration. AB - The motogenic activity of migration-stimulating factor, a truncated isoform of fibronectin (FN), has been attributed to the IGD motifs present in its FN type 1 modules. The structure-function relationship of various recombinant IGD containing FN fragments is now investigated. Their structure is assessed by solution state NMR and their motogenic ability tested on fibroblasts. Even conservative mutations in the IGD motif are inactive or have severely reduced potency, while the structure remains essentially the same. A fragment with two IGD motifs is 100 times more active than a fragment with one and up to 10(6) times more than synthetic tetrapeptides. The wide range of potency in different contexts is discussed in terms of cryptic FN sites and cooperativity. These results give new insight into the stimulation of fibroblast migration by IGD motifs in FN. PMID- 17921137 TI - Pivotal involvement of Fcgamma receptor IIA in the neutralization of lipopolysaccharide signaling via a potent novel anti-TLR4 monoclonal antibody 15C1. AB - The mammalian Toll-like receptor (TLR) family has evolved to sense pathogens in the environment and protect the host against infection. TLR4 recognizes lipopolysaccharide (LPS) from Gram-negative bacteria and induces a signaling cascade that, when exaggerated, has been associated with severe sepsis. We have generated a TLR4-specific monoclonal antibody, 15C1, which neutralizes LPS induced TLR4 activation in a dose-dependent manner. 15C1 potently blocks the effects of LPS on a panel of primary cells and cell lines in vitro. The binding of 15C1 was mapped to an epitope in the second portion of the extracellular region of TLR4, which has been shown previously to be functionally important in the recognition of LPS. Furthermore, we demonstrate a novel mechanism of inhibition, as the effects of 15C1 are partially Fc-dependent, involving the regulatory Fcgamma receptor IIA (CD32A). In addition to introducing 15C1 as a potent clinical candidate for use in the treatment of LPS-mediated indications, our work demonstrates a newly discovered pathway whose manipulation is pivotal in achieving optimal neutralizing benefit. PMID- 17921138 TI - Regulation of peroxiredoxins by nitric oxide in immunostimulated macrophages. AB - Reactive oxygen species and nitric oxide (NO) are capable of both mediating redox sensitive signal transduction and eliciting cell injury. The interplay between these messengers is quite complex, and intersection of their signaling pathways as well as regulation of their fluxes requires tight control. In this regard, peroxiredoxins (Prxs), a recently identified family of six thiol peroxidases, are central because they reduce H2O2, organic peroxides, and peroxynitrite. Here we provide evidence that endogenously produced NO participates in protection of murine primary macrophages against oxidative and nitrosative stress by inducing Prx I and VI expression at mRNA and protein levels. We also show that NO prevented the sulfinylation-dependent inactivation of 2-Cys Prxs, a reversible overoxidation that controls H2O2 signaling. In addition, studies using macrophages from sulfiredoxin (Srx)-deficient mice indicated that regeneration of 2-Cys Prxs to the active form was dependent on Srx. Last, we show that NO increased Srx expression and hastened Srx-dependent recovery of 2-Cys Prxs. We therefore propose that modulation by NO of Prx expression and redox state, as well as up-regulation of Srx expression, constitutes a novel pathway that contributes to antioxidant response and control of H2O2-mediated signal transduction in mammals. PMID- 17921140 TI - Profiling the enzymatic properties and inhibition of human complement factor B. AB - Human complement factor B is the crucial catalytic component of the C3 convertase enzyme that activates the alternative pathway of complement-mediated immunity. Although a serine protease in its own right, factor B circulates in human serum as an inactive zymogen and there is a crystal structure only for the inactive state of factor B and various fragments. To provide greater insight to the catalytic function and properties of factor B, we have used short para nitroanilide derivatives of 4- to 15-residue peptides as substrates to profile the catalytic properties of factor B. Among factors found to influence catalytic activity of factor B was an unusual dependence on pH. Non-physiological alkaline conditions strongly promoted substrate cleavage by factor B, consistent with a pH accessible conformation of the enzyme that may be critical for catalytic function. Small N-terminal extensions to conventional hexapeptide para nitroanilide substrates significantly increased catalytic activity of factor B, which was more selective for its cleavage site than trypsin. The new chromogenic assay enabled optimization of catalysis conditions, the profiling of different substrate sequences, and the development of the first reversible and competitive substrate-based inhibitor of factor B. The inhibitor was also shown to prevent in vitro formation of C3a from C3 by factor B, by synthetic and by natural C3 convertase of the alternative complement activation pathway, and to block formation of membrane attack complex. The availability of a reversible substrate based inhibitor that could stabilize the active conformation of factor B, in conjunction with a pH-promoted higher processing activity, may offer a new avenue to obtain crystal structures of factor B and C3 convertase in an active conformation. PMID- 17921139 TI - Evidence for physical interaction between the immunoglobulin heavy chain variable region and the 3' regulatory region. AB - B cell-specific expression of immunoglobulin heavy chain (IgH) genes utilizes two cis regulatory regions, the intronic enhancer (Emicro), located in the J(H) Cmicro intron, and a complex regulatory region that lies 3' to the IgH gene cluster, 3' RR. We hypothesized that the 3' RR is involved in IgH gene transcription in plasma cells via physical interaction between distal 3' RR enhancers and target V(H) sequences, with loop formation by intervening DNA. In support of this hypothesis we report sequence data at DNA recombination breakpoints as evidence for loop formation preceding DNA inversion in a plasma cell line. In addition, using the chromosome conformation capture technique, physical interactions between V(H) and 3' RR were analyzed directly and detected in MPC11 plasma cells and variants and normal splenic B cells but not detected in splenic T cells or in non-B cells. V(H)-3' RR interactions were present in the absence of Emicro, but when the hs1,2 enhancer was replaced by a Neo(R) gene in a variant cell line lacking Emicro, H chain expression was lost, and interactions between V(H) and 3' RR and among the 3' RR regulators themselves were severely disrupted. In addition, the chromosome conformation capture technique detected interactions between the myc promoter and 3' RR elements in MPC11, which like other plasmacytomas contains a reciprocal translocation between the c-myc and the IgH locus. In sum, our data support a hypothesis that cis V(H)-3' RR and myc-3' RR interactions involve physical interactions between these DNA elements. PMID- 17921141 TI - Selection of genetically encoded fluorescent single domain antibodies engineered for efficient expression in Escherichia coli. AB - Genetically encoded fluorescent antibodies are desirable for many applications in biotechnology and proteomics. However, the efficient production of single chain antibodies fused to fluorescent proteins like green fluorescent protein is still a major challenge. This is due to the opposite redox requirements of recombinant antibodies and fluorescent proteins for proper folding. To overcome this obstacle, we designed a novel strategy to directly select functional fluorescent antibodies (fluobodies) engineered for efficient cytosolic expression in Escherichia coli, starting from a llama heavy chain antibody (VHH) library. We used this strategy to produce several VHH fluobodies directed against microtubules. After biochemical characterization in vitro, we used a selected fluobody as a convenient biomarker of the microtubule cytoskeleton in eukaryotic cells and engineered its properties. This work provides a reliable approach for the production of fluobodies in wild-type E. coli and furthermore suggests that fusions of recombinant VHH with other cytosolic proteins are now possible, creating new opportunities for biotechnology and biomedical research. PMID- 17921142 TI - Spectroscopic and kinetic studies of Y114F and W116F mutants of Me2SO reductase from Rhodobacter capsulatus. AB - Mutants of the active site residues Trp-116 and Tyr-114 of the molybdenum containing Me(2)SO reductase from Rhodobacter capsulatus have been examined spectroscopically and kinetically. The Y114F mutant has an increased rate constant for oxygen atom transfer from Me(2)SO to reduced enzyme, the result of lower stability of the E(red).Me(2)SO complex. The absorption spectrum of this species (but not that of either oxidized or reduced enzyme) is significantly perturbed in the mutant relative to wild-type enzyme, consistent with Tyr-114 interacting with bound Me(2)SO. The as-isolated W116F mutant is only five coordinate, with one of the two equivalents of the pyranopterin cofactor found in the enzyme dissociated from the molybdenum and replaced by a second Mo=O group. Reduction of the mutant with sodium dithionite and reoxidation with Me(2)SO, however, regenerates the long-wavelength absorbance of functional enzyme, although the wavelength maximum is shifted to 670 nm from the 720 nm of wild-type enzyme. This "redox-cycled" mutant exhibits a Me(2)SO reducing activity and overall reaction mechanism similar to that of wild-type enzyme but rapidly reverts to the inactive five-coordinate form in the course of turnover. PMID- 17921143 TI - Protein-tyrosine phosphatase H1 controls growth hormone receptor signaling and systemic growth. AB - Several protein-tyrosine phosphatases (PTPs) have been implicated in the control of growth hormone receptor (GHR) signaling, but none have been shown to affect growth in vivo. We have applied a battery of molecular and cellular approaches to test a family-wide panel of PTPs for interference with GHR signaling. Among the subset of PTPs that showed activity in multiple readouts, we selected PTP H1/PTPN3 for further in vivo studies and found that mice lacking the PTP-H1 catalytic domain show significantly enhanced growth over their wild type littermates. In addition, PTP-H1 mutant animals had enhanced plasma and liver mRNA expression of insulin-like growth factor 1, as well as increased bone density and mineral content. These observations point to a controlling role for PTP-H1 in modulating GHR signaling and systemic growth through insulin-like growth factor 1 secretion. PMID- 17921144 TI - Microtubule motors regulate ISOC activation necessary to increase endothelial cell permeability. AB - Calcium store depletion activates multiple ion channels, including calcium selective and nonselective channels. Endothelial cells express TRPC1 and TRPC4 proteins that contribute to a calcium-selective store-operated current, I(SOC). Whereas thapsigargin activates the I(SOC) in pulmonary artery endothelial cells (PAECs), it does not activate I(SOC) in pulmonary microvascular endothelial cells (PMVECs), despite inducing a significant rise in global cytosolic calcium. Endoplasmic reticulum exhibits retrograde distribution in PMVECs when compared with PAECs. We therefore sought to determine whether endoplasmic reticulum-to plasma membrane coupling represents an important determinant of I(SOC) activation in PAECs and PMVECs. Endoplasmic reticulum organization is controlled by microtubules, because nocodozole induced microtubule disassembly and caused retrograde endoplasmic reticulum collapse in PMVECs. In PMVECs, rolipram treatment produced anterograde endoplasmic reticulum distribution and revealed a thapsigargin-activated I(SOC) that was abolished by nocodozole and taxol. Microtubule motors control organelle distribution along microtubule tracks, with the dynein motor causing retrograde movement and the kinesin motor causing anterograde movement. Dynamitin expression reduces dynein motor function inducing anterograde endoplasmic reticulum transport, which allows for direct activation of I(SOC) by thapsigargin in PMVECs. In contrast, expression of dominant negative kinesin light chain reduces kinesin motor function and induces retrograde endoplasmic reticulum transport; dominant negative kinesin light chain expression prevented the direct activation of I(SOC) by thapsigargin in PAECs. I(SOC) activation is an important step leading to disruption of cell-cell adhesion and increased macromolecular permeability. Thus, microtubule motor function plays an essential role in activating cytosolic calcium transitions through the membrane I(SOC) channel leading to endothelial barrier disruption. PMID- 17921145 TI - Transcription factor E is a part of transcription elongation complexes. AB - A homologue of the N-terminal domain of the alpha subunit of the general eukaryotic transcription factor TFE is encoded in the genomes of all sequenced archaea, but the position of archaeal TFE in transcription complexes has not yet been defined. We show here that TFE binds nonspecifically to single-stranded DNA, and photochemical cross-linking revealed TFE binding to a preformed open transcription bubble. In preinitiation complexes, the N-terminal part of TFE containing a winged helix-turn-helix motif is cross-linked specifically to DNA of the nontemplate DNA strand at positions -9 and -11. In complexes stalled at +20, TFE cross-linked specifically to positions +9, +11, and +16 of the non-template strand. Analyses of transcription complexes stalled at position +20 revealed a TFE-dependent increase of the resumption efficiency of stalled RNA polymerase and a TFE-induced enhanced permanganate sensitivity of thymine residues in the transcription bubble. These results demonstrate the presence of TFE in early elongation complexes and suggest a role of TFE in stabilization of the transcription bubble during elongation. PMID- 17921146 TI - The serine protease domain of hepatitis C viral NS3 activates RNA helicase activity by promoting the binding of RNA substrate. AB - Nonstructural (NS) protein 3 is a DEXH/D-box motor protein that is an essential component of the hepatitis C viral (HCV) replicative complex. The full-length NS3 protein contains two functional modules, both of which are essential in the life cycle of HCV: a serine protease domain at the N terminus and an ATPase/helicase domain (NS3hel) at the C terminus. Truncated NS3hel constructs have been studied extensively; the ATPase, nucleic acid binding, and helicase activities have been examined and NS3hel has been used as a target in the development of antivirals. However, a comprehensive comparison of NS3 and NS3hel activities has not been performed, so it remains unclear whether the protease domain plays a vital role in NS3 helicase function. Given that many DEXH/D-box proteins are activated upon interaction with cofactor proteins, it is important to establish if the protease domain acts as the cofactor for stimulating NS3 helicase function. Here we show that the protease domain greatly enhances both the direct and functional binding of RNA to NS3. Whereas electrostatics plays an important role in this process, there is a specific allosteric contribution from the interaction interface between NS3hel and the protease domain. Most importantly, we establish that the protease domain is required for RNA unwinding by NS3. Our results suggest that, in addition to its role in cleavage of host and viral proteins, the NS3 protease domain is essential for the process of viral RNA replication and, given its electrostatic contribution to RNA binding, it may also assist in packaging of the viral RNA. PMID- 17921147 TI - Aciniform spidroin, a constituent of egg case sacs and wrapping silk fibers from the black widow spider Latrodectus hesperus. AB - Spiders produce high performance fibers with diverse mechanical properties and biological functions. Molecular and biochemical studies of spider egg case silk have revealed that the main constituent of the large diameter fiber contains the fibroin TuSp1. Here we demonstrate by SDS-PAGE and protein silver staining the presence of a distinct approximately 300-kDa polypeptide that is found in solubilized egg case sacs. Combining matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry and reverse genetics, we have isolated a novel gene called AcSp1-like and demonstrate that its protein product is assembled into the small diameter fibers of egg case sacs and wrapping silks from the black widow spider, Latrodectus hesperus. BLAST searches of the NCBInr protein data base using the amino acid sequence of AcSp1-like revealed similarity to AcSp1, an inferred protein proposed to be a component of wrapping silk. However, the AcSp1-like protein was found to display more nonuniformity in its internal iterated repeat modules than the putative AcSp1 fibroin. Real time quantitative PCR analysis demonstrates that the AcSp1-like gene displays an aciniform gland-restricted pattern of expression. The amino acid composition of the fibroins extracted from the luminal contents of the aciniform glands was remarkably similar to the predicted amino acid composition of the AcSp1-like protein, which supports the assertion that AcSp1-like protein represents the major constituent stored within the aciniform gland. Collectively, our findings provide the first direct molecular evidence for the involvement of the aciniform gland in the production of a common fibroin that is assembled into the small diameter threads of egg case and wrapping silk of cob weavers. PMID- 17921148 TI - Human DNA mismatch repair: coupling of mismatch recognition to strand-specific excision. AB - Eukaryotic mismatch-repair (MMR) proteins MutSalpha and MutLalpha couple recognition of base mismatches to strand-specific excision, initiated in vivo at growing 3' ends and 5' Okazaki-fragment ends or, in human nuclear extracts, at nicks in exogenous circular substrates. We addressed five biochemical questions relevant to coupling models. Excision remained fully efficient at DNA:MutSalpha ratios of nearly 1 to 1 at various mismatch-nick distances, suggesting a requirement for only one MutSalpha molecule per substrate. As the mismatch-nick DNA contour distance D in exogenous substrates increased from 0.26 to 0.98 kbp, initiation of excision in extracts decreased as D(-0.43) rather than the D(-1) to D(-2) predicted by some translocation or diffusion models. Virtually all excision was along the shorter (3'-5') nick-mismatch, even when the other (5'-3') path was less than twice as long. These observations argue against stochastically directed translocating/diffusing recognition complexes. The failure of mismatched DNA in trans to provoke excision of separate nicked homoduplexes argues against one stage (concerted) triggering of excision initiation by recognition complexes acting through space. However, proteins associated with gapped DNA did appear to compete in trans with those in cis to mismatch-associated proteins. Thus, as in Escherichia coli, eukaryotic MMR may involve distinct initial-activation and excision-path-commitment stages. PMID- 17921149 TI - Binding of chara Myosin globular tail domain to phospholipid vesicles. AB - Binding of Chara myosin globular tail domain to phospholipid vesicles was investigated quantitatively. It was found that the globular tail domain binds to vesicles made from acidic phospholipids but not to those made from neutral phospholipids. This binding was weakened at high KCl concentration, suggesting that the binding is electrostatic by nature. The dissociation constant for the binding of the globular tail domain to 20% phosphatidylserine vesicles (similar to endoplasmic reticulum in acidic phospholipid contents) at 150 mM KCl was 273 nM. The free energy change due to this binding calculated from the dissociation constant was -37.3 kJ mol(-1). Thus the bond between the globular tail domain and membrane phospholipids would not be broken when the motor domain of Chara myosin moves along the actin filament using the energy of ATP hydrolysis (DeltaG degrees ' = -30.5 kJ mol(-1)). Our results suggested that direct binding of Chara myosin to the endoplasmic reticulum membrane through the globular tail domain could work satisfactorily in Chara cytoplasmic streaming. We also suggest a possible regulatory mechanism of cytoplasmic streaming including phosphorylation-dependent dissociation of the globular tail domain from the endoplasmic reticulum membrane. PMID- 17921150 TI - 2-Hydroxyisoflavanone dehydratase is a critical determinant of isoflavone productivity in hairy root cultures of Lotus japonicus. AB - Hairy root cultures of a model legume, Lotus japonicus, were established to characterize two heterologous cDNAs encoding enzymes involved in isoflavone biosynthesis, i.e. licorice 2-hydroxyisoflavanone synthase (IFS) and soybean 2 hydroxyisoflavanone dehydratase (HID) catalyzing sequential reactions to yield isoflavones. While the control and the IFS overexpressor did not accumulate detectable isoflavones, the HID overexpressors did accumulate daidzein and genistein, showing that HID is a critical determinant of isoflavone productivity. Production of coumestrol in all the genotypes and isoliquiritigenin/liquiritigenin in IFS + HID-overexpressing lines was also noted. These results provide insight into the regulatory mechanism that controls isoflavonoid biosynthesis. PMID- 17921152 TI - The long and winding road: influences of intracellular metabolite diffusion on cellular organization and metabolism in skeletal muscle. AB - A fundamental principle of physiology is that cells are small in order to minimize diffusion distances for O(2) and intracellular metabolites. In skeletal muscle, it has long been recognized that aerobic fibers that are used for steady state locomotion tend to be smaller than anaerobic fibers that are used for burst movements. This tendency reflects the interaction between diffusion distances and aerobic ATP turnover rates, since maximal intracellular diffusion distances are ultimately limited by fiber size. The effect of diffusion distance on O(2) flux in muscle has been the subject of quantitative analyses for a century, but the influence of ATP diffusion from mitochondria to cellular ATPases on aerobic metabolism has received much less attention. The application of reaction diffusion mathematical models to experimental measurements of aerobic metabolic processes has revealed that the extreme diffusion distances between mitochondria found in some muscle fibers do not necessarily limit the rates of aerobic processes per se, as long as the metabolic process is sufficiently slow. However, skeletal muscle fibers from a variety of animals appear to have intracellular diffusion distances and/or fiber sizes that put them on the brink of diffusion limitation. Thus, intracellular metabolite diffusion likely influences the evolution of muscle design and places limits on muscle function. PMID- 17921151 TI - Drug shop regulation and malaria treatment in Tanzania--why do shops break the rules, and does it matter? AB - Regulatory infringements are extremely common in low-income countries, especially with respect to retail pharmaceutical sales. There have been few practical suggestions on public policy responses other than stricter regulatory enforcement, which governments are often unable, or unwilling, to do. This paper explores the challenges of regulating retail drug sellers, and potential solutions, through a case study of malaria treatment in rural Tanzania where small drug shops are a common source of medicine. Infringement of health-related regulation was extremely common. Most stores lacked valid permits, and illegal stocking of prescription-only medicines and unpackaged tablets was the norm. Most stocked unregistered drugs, and no serving staff met the qualification requirements. Infringements are likely to have reflected infrequent regulatory inspections, a failure of regulatory authorities to implement sanctions, successful concealment of regulatory violations, and the tacit permission of local regulatory staff. Eliminating regulatory infringements is unlikely to be feasible, and could be undesirable if access to essential medicines is reduced. Alternatives include bringing official drug regulation closer into line with locally legitimate practices; greater use of positive incentives for providers; and consumer involvement. Such a change in approach has the potential to provide a firmer platform for public-private collaboration to improve shop-based treatment. PMID- 17921153 TI - Reappraisal of the comparative cost of human locomotion using gait-specific allometric analyses. AB - The alleged high net energy cost of running and low net energy cost of walking in humans have played an important role in the interpretation of the evolution of human bipedalism and the biomechanical determinants of the metabolic cost of locomotion. This study re-explores how the net metabolic energy cost of running and walking (J kg(-1) m(-1)) in humans compares to that of animals of similar mass using new allometric analyses of previously published data. Firstly, this study shows that the use of the slope of the regression between the rate of energy expenditure and speed to calculate the net energy cost of locomotion overestimates the net cost of human running. Also, the net energy cost of human running is only 17% higher than that predicted based on their mass. This value is not exceptional given that over a quarter of the previously examined mammals and birds have a net energy cost of running that is 17% or more above their allometrically predicted value. Using a new allometric equation for the net energy cost of walking, this study also shows that human walking is 20% less expensive than predicted for their mass. Of the animals used to generate this equation, 25% have a relatively lower net cost of walking compared with their allometrically predicted value. This new walking allometric analysis also indicates that the scaling of the net energy cost of locomotion with body mass is gait dependent. In conclusion, the net costs of running and walking in humans are moderately different from those predicted from allometry and are not remarkable for an animal of its size. PMID- 17921155 TI - The acoustic mechanics of stick slip friction in the California spiny lobster (Panulirus interruptus). AB - The dynamic interplay between static and sliding friction is fundamental to many animal movements. One interesting example of stick-slip friction is found in the sound-producing apparatus of many spiny lobster species (Palinuridae). The acoustic movements of the spiny lobster's plectrum over the file are generated by stick-slip friction between the two surfaces. We examined the microscopic anatomy, kinematics, acoustics and frictional properties of the California spiny lobster (Panulirus interruptus) toward the goal of quantitatively characterizing the frictional and acoustic mechanics of this system. Using synchronous high speed video and sound recordings, we tested whether plectrum kinematics are correlated with acoustic signal features and found that plectrum velocity is positively correlated with acoustic amplitude. To characterize the frictional mechanics of the system, we measured frictional forces during sound production using excised plectrums and files. Similar to rubber materials sliding against hard surfaces, the static coefficient of friction in this system was on average 1.7. The change in the coefficient of friction across each stick-slip cycle varied substantially with an average change of 1.1. Although driven at a constant speed, the plectrum slipped at velocities that were positively correlated with the normal force between the two surfaces. Studies of friction in biological systems have focused primarily on adhesion and movement, while studies of stick slip acoustics have remained under the purview of musical acoustics and engineering design. The present study offers an integrative analysis of an unusual bioacoustic mechanism and contrasts its physical parameters with other biological and engineered systems. PMID- 17921154 TI - Search for hepatopancreatic ecdysteroid-responsive genes during the crayfish molt cycle: from a single gene to multigenicity. AB - The expression of the vitellogenin gene of the red-claw crayfish Cherax quadricarinatus (CqVg) was previously demonstrated in male crayfish during an endocrinologically induced molt cycle. The hypothesis that this expression is under the direct control of ecdysteroids was tested in this study both in vivo and in vitro. Unlike vitellogenin of insects, CqVg was not found to be ecdysteroid-responsive. Thus, a multigenic approach was employed for the identification of other hepatopancreatic ecdysteroid-responsive genes by a cDNA microarray. For the purposes of this study, a multi-parametric molt-staging technique, based on X-ray detection of gastrolith growth, was developed. To identify ecdysteroid-responsive genes during premolt, the molt cycle was induced by two manipulations, 20-hydroxyecdysone administration and X-organ-sinus gland complex removal; both resulted in significant elevation of ecdysteroids. Two clusters of affected genes (129 and 122 genes, respectively) were revealed by the microarray. It is suggested that only genes belonging to similarly responsive (up or downregulated) gene clusters in both manipulations (102 genes) could be considered putative ecdysteroid-responsive genes. Some of these ecdysteroid responsive genes showed homology to genes controlling chitin metabolism, proteases and other cellular activities, while 56.8% were unknown. The majority of the genes were downregulated, presumably by an energetic shift of the hepatopancreas prior to ecdysis. The effect of 20-hydroxyecdysone on representative genes from this group was confirmed in vitro using a hepatopancreas tissue culture. This approach for ecdysteroid-responsive gene identification could also be implemented in other tissues for the elucidation of ecdysteroid-specific signaling pathways during the crustacean molt cycle. PMID- 17921156 TI - Natural variation in food acquisition mediated via a Drosophila cGMP-dependent protein kinase. AB - In natural environments where food abundance and quality can change drastically over time, animals must continuously alter their food acquisition strategies. Although genetic variation contributes to this plasticity, the specific genes involved and their interactions with the environment are poorly understood. Here we report that natural variation in the Drosophila gene, foraging (for), which encodes a cGMP-dependent protein kinase (PKG), affects larval food acquisition in an environmentally dependent fashion. When food is plentiful, the wild-type rover (for(R)) allele confers lower food intake and higher glucose absorption than both the wild-type sitter (for(s)) allele and the mutant for(s2) allele. When food is scarce, for(R), for(s) and for(s2) larvae increase food intake to a common maximal level, but for(R) larvae retain their increased absorption efficiency. Changes in for expression can induce corrective behavioral modifications in response to food deprivation. When reared in environments with low food levels, for(R) larvae have higher survivorship and faster development than for(s) and for(s2) larvae. Together, these results show that natural variation in for has far reaching implications affecting a suite of phenotypes involved in the regulation of food acquisition. PMID- 17921157 TI - Circadian timed episodic-like memory - a bee knows what to do when, and also where. AB - This study investigates how the colour, shape and location of patterns could be memorized within a time frame. Bees were trained to visit two Y-mazes, one of which presented yellow vertical (rewarded) versus horizontal (non-rewarded) gratings at one site in the morning, while another presented blue horizontal (rewarded) versus vertical (non-rewarded) gratings at another site in the afternoon. The bees could perform well in the learning tests and various transfer tests, in which (i) all contextual cues from the learning test were present; (ii) the colour cues of the visual patterns were removed, but the location cue, the orientation of the visual patterns and the temporal cue still existed; (iii) the location cue was removed, but other contextual cues, i.e. the colour and orientation of the visual patterns and the temporal cue still existed; (iv) the location cue and the orientation cue of the visual patterns were removed, but the colour cue and temporal cue still existed; (v) the location cue, and the colour cue of the visual patterns were removed, but the orientation cue and the temporal cue still existed. The results reveal that the honeybee can recall the memory of the correct visual patterns by using spatial and/or temporal information. The relative importance of different contextual cues is compared and discussed. The bees' ability to integrate elements of circadian time, place and visual stimuli is akin to episodic-like memory; we have therefore named this kind of memory circadian timed episodic-like memory. PMID- 17921158 TI - Environmental stress affects the expression of a carotenoid-based sexual trait in male zebra finches. AB - Abiotic factors including thermal stress are suggested to exert constrains on sexual ornaments through trade-offs between sexual displays and physiological functions related to self-maintenance. Given the health properties of carotenoid pigments, carotenoid-based ornaments offer a relevant context in which to investigate the effect of environmental stress, such as ambient temperature, on the production and maintenance of secondary sexual traits and, also, to explore the proximate mechanisms shaping their expression. In this study, we exposed male zebra finches (Taeniopygia guttata) to environmental stress by exposing them to two temperature regimes (6 and 26 degrees C) over a 4 week period. Simultaneously, half of the males in each temperature group were supplemented with carotenoids, whereas the other half were not. The expression of a carotenoid based sexual trait (bill colour) and the amount of circulating carotenoids were assessed before and at the end of the experiment. Carotenoid-supplemented males developed a redder bill, but the effect of supplementation was reduced under cold exposure. However, we found evidence that birds facing a cold stress were carotenoid limited, since supplemented males developed redder bills than the non supplemented ones. Interestingly, while cold-exposed and non-supplemented males developed duller bills, they circulated a higher amount of carotenoids at the end of the experiment compared to the pre-experimental values. Together, these results suggest that ambient temperature might contribute to the modulation of the expression of carotenoid-based ornaments. Our findings suggest that carotenoids are a limiting resource under cold exposure and that they might be prioritized for self-maintenance at the expense of the ornament. The physiological functions related to self-maintenance that might have benefited from carotenoid saving are discussed. PMID- 17921159 TI - Kinematics of jumping in leafhopper insects (Hemiptera, Auchenorrhyncha, Cicadellidae). AB - The jumping movements and performance of leafhopper insects (Hemiptera, Auchenorrhyncha, Cicadellidae) were analysed from high-speed sequences of images captured at rates up to 5000 frames s(-1). The propulsion for a jump was delivered by rapid and synchronous movements of the hind legs that are twice the length of the other legs, almost as long as the body, and represent 3.8% of the body mass. The wings were not moved before take-off, but the jump frequently launched a flight. The front and middle legs set the attitude of the body in preparation for a jump but were usually raised from the ground before take-off. The movements of the hind legs occurred in three distinct phases. First, a levation phase of 15-30 ms, in which both hind legs were moved forward and medially so that they were positioned directly beneath the body with their tibio tarsal joints pressed against each other. Second, a holding phase lasting 10-200 ms, in which the hind legs remained stationary in the fully levated position. Third, a rapid jump phase, in which both hind legs were simultaneously depressed about their coxo-trochanteral joints and extended at their femoro-tibial joints. This phase lasted 5-6 ms on average, with the fastest movements accomplished in 2.75 ms and involving rotations of the coxo-trochanteral joints of 44,000 deg. s( 1). In the best jumps by Aphrodes, a peak take-off velocity of 2.9 m s(-1) was achieved by an acceleration of 1055 m s(-2), equivalent to 108 times gravity. This jumping performance required an energy output of 77 microJ, a power output of 28 mW and exerted a force of 19 mN, or 100 times its body mass. PMID- 17921160 TI - Anatomy of the hind legs and actions of their muscles during jumping in leafhopper insects. AB - The rapid and simultaneous depression of the trochantera about the coxae of both hind legs of leafhoppers are the key joint movements powering a jump. The present study analyses the structure of these joints and the actions of the muscles that move them. The hind coxae are huge and are linked to each other at the midline by a protrusion from one coxa that inserts in a socket of the other and acts like a press-stud (popper) fastener. This asymmetry is not reflected in any left- or right-handed preference either within one species or between species. The movements of the joints in a jump are monitored by a number of possible proprioceptors that should be activated when a hind leg is fully levated in preparation for a jump: a hair row and two hair plates on the coxa, a hair plate on a trochanteral pivot with a coxa, and femoral spines at the femoro-tibial joint. The depressor and levator muscles that move the trochanter are of similar size and together occupy the greater part of the metathorax. Their lever arms are similar when the leg is fully levated, but the lever arm of the depressor increases with initial depression of the coxo-trochanteral joint while that of the levator declines. A jump is preceded by activity in the trochanteral depressor and levator muscles, which results in a forward movement of the coxa and metathorax with the trochanter fully levated. This period of co-contraction could result in storage of energy in skeletal structures in the thorax. Just before the rapid depression of the trochanter in the jump movement the frequency of depressor spikes increases while that in the levator declines, releasing any force stored by the preceding muscle contractions. These bursts of depressor spikes occur at the same time in the left and right muscles but none of the individual motor spikes appeared to be synchronous on the two sides. PMID- 17921161 TI - Male accessory glands of Drosophila melanogaster make a secreted angiotensin I converting enzyme (ANCE), suggesting a role for the peptide-processing enzyme in seminal fluid. AB - Angiotensin I-converting enzyme (ACE) expressed on the surface of endothelial cells is responsible for the last step in the synthesis of circulating angiotensin II and the inactivation of bradykinin. Mammalian ACE is also expressed in the prostate with other components of the renin-angiotensin system, and in developing spermatids, where the peptidase activity is known to be critical for normal sperm function. The importance of an ACE gene to male fertility has also been demonstrated in Drosophila melanogaster, where Ance is expressed in spermatids, and hypomorphic alleles of Ance cause a defect in spermiogenesis. Here we show that ANCE, which shares many enzymatic properties with mammalian ACE, is also a product of the male accessory gland of D. melanogaster. It is expressed in the secondary cells and is associated with the electron dense granule within the large vesicles of these cells. ACE proteolytic activity is lost from the accessory glands during mating, consistent with transfer to the mated female in the seminal fluid. The accessory gland ACE-like activity might have an evolutionarily conserved function processing biologically active peptides with a role in male fertility. PMID- 17921162 TI - The sensory basis of roost finding in a forest bat, Nyctalus noctula. AB - Tree cavities are a critical resource for most forest-dwelling bats. Yet, it is not known how bats search for new sites and, in particular, find entrances to cavities. Here, we evaluated the importance of different sensory channels for the detection of tree roosts by the noctule bat Nyctalus noctula. Specifically, we tested the role of three non-social cues (echo information, visual information and temperature-related cues) and two social sensory cues (conspecific echolocation calls and the presence of bat olfactory cues). We set up an experiment in a flight room that mimicked natural conditions. In the flight room, we trained wild-caught bats kept in captivity for a short while to find the entrance to an artificial tree cavity. We measured the bats' hole-finding performance based on echolocation cues alone and then presented the bat with one of four additional sensory cues. Our data show that conspecific echolocation calls clearly improved the bats' performance in finding tree holes, both from flying (long-range detection) and when they were crawling on the trunk (short range detection). The other cues we presented had no, or only weak, effects on performance, implying that detection of new cavities from a distance is difficult for noctules if no additional social cues, in particular calls from conspecifics, are present. We conclude that sensory constraints strongly limit the effectiveness of finding new cavities and may in turn promote sociality and acoustic information transfer among bats. As acoustic cues clearly increased the bats' detection performance, we suggest that eavesdropping is an important mechanism for reducing the costs of finding suitable roosts. PMID- 17921163 TI - Visually guided obstacle avoidance in the box jellyfish Tripedalia cystophora and Chiropsella bronzie. AB - Box jellyfish, cubomedusae, possess an impressive total of 24 eyes of four morphologically different types. Two of these eye types, called the upper and lower lens eyes, are camera-type eyes with spherical fish-like lenses. Compared with other cnidarians, cubomedusae also have an elaborate behavioral repertoire, which seems to be predominantly visually guided. Still, positive phototaxis is the only behavior described so far that is likely to be correlated with the eyes. We have explored the obstacle avoidance response of the Caribbean species Tripedalia cystophora and the Australian species Chiropsella bronzie in a flow chamber. Our results show that obstacle avoidance is visually guided. Avoidance behavior is triggered when the obstacle takes up a certain angle in the visual field. The results do not allow conclusions on whether color vision is involved but the strength of the response had a tendency to follow the intensity contrast between the obstacle and the surroundings (chamber walls). In the flow chamber Tripedalia cystophora displayed a stronger obstacle avoidance response than Chiropsella bronzie since they had less contact with the obstacles. This seems to follow differences in their habitats. PMID- 17921164 TI - Spectral and spatial properties of polarized light reflections from the arms of squid (Loligo pealeii) and cuttlefish (Sepia officinalis L.). AB - On every arm of cuttlefish and squid there is a stripe of high-reflectance iridophores that reflects highly polarized light. Since cephalopods possess polarization vision, it has been hypothesized that these polarized stripes could serve an intraspecific communication function. We determined how polarization changes when these boneless arms move. By measuring the spectral and polarizing properties of the reflected light from samples at various angles of tilt and rotation, we found that the actual posture of the arm has little or no effect on partial polarization or the e-vector angle of the reflected light. However, when the illumination angle changed, the partial polarization of the reflected light also changed. The spectral reflections of the signals were also affected by the angle of illumination but not by the orientation of the sample. Electron microscope samples showed that these stripes are composed of several groups of multilayer platelets within the iridophores. The surface normal to each group is oriented at a different angle, which produces essentially constant reflection of polarized light over a range of viewing angles. These results demonstrate that cuttlefish and squid could send out reliable polarization signals to a receiver regardless of arm orientation. PMID- 17921165 TI - A chymotrypsin-like serine protease interacts with the chitin synthase from the midgut of the tobacco hornworm. AB - The chitin portion of the peritrophic matrix in the midgut of the tobacco hornworm, Manduca sexta, is produced by chitin synthase 2 (CHS2), a transmembrane family II glycosyltransferase, located at the apical tips of brush border microvilli. To look for proteins that potentially interact with CHS2, we performed yeast two-hybrid screening, identifying a novel chymotrypsin-like protease (CTLP1) that binds to the extracellular carboxyterminal domain of CHS2. The occurrence of this interaction in vivo is supported by co-localization and co immunoprecipitation data. Based on our findings we propose that chitin synthesis is controlled by an intestinal proteolytic signalling cascade linking chitin synthase activity to the nutritional state of the larvae. PMID- 17921166 TI - Sex and flow: the consequences of fluid shear for sperm-egg interactions. AB - Fertilization is a complex interaction among biological traits of gametes and physical properties of the fluid environment. At the scale of fertilization (0.01 1 mm), sperm encounter eggs while being transported within a laminar (or viscous) shear flow. Varying laminar-shear in a Taylor-Couette flow tank, our experiments simulated important aspects of small-scale turbulence within the natural habitats of red abalone (Haliotis rufescens), a large marine mollusk and external fertilizer. Behavioral interactions between individual cells, sperm-egg encounter rates, and fertilization success were quantified, simultaneously, using a custom built infrared laser and computer-assisted video imaging system. Relative to still water, sperm swam faster and moved towards an egg surface, but only in comparatively slow flows. Encounter rate, swim speed and orientation, and fertilization success each peaked at the lowest shear tested (0.1 s(-1)), and then decayed as shear increased beyond 1.0 s(-1). The decay did not result, however, from damage to either sperm or eggs. Analytical and numerical models were used to estimate the propulsive force generated by sperm swimming (F(swim)) and the shear force produced by fluid motion within the vicinity of a rotating egg (F(shear)). To first order, male gametes were modeled as prolate spheroids. The ratio F(swim)/F(shear) was useful in explaining sperm-egg interactions. At low shears where F(swim)/F(shear)>1, sperm swam towards eggs, encounter rates were pronounced, and fertilization success was very high; behavior overpowered fluid motion. In contrast, sperm swimming, encounter rate and fertilization success all decayed rapidly when F(swim)/F(shear)<1; fluid motion dominated behavior. The shears maximizing fertilization success in the lab typically characterized natural flow microenvironments of spawning red abalone. Gamete behavior thus emerges as a critical determinant of sexual reproduction in the turbulent sea. PMID- 17921167 TI - Phonotactic walking paths of field crickets in closed-loop conditions and their simulation using a stochastic model. AB - Field cricket females localize one of many singing males in the field in closed loop multi-source conditions. To understand this behaviour, field cricket phonotaxis was investigated in a closed-loop walking phonotaxis paradigm, in response to two simultaneously active speakers playing aphasic calling songs. Female phonotactic paths were oriented towards the louder sound sources, but showed great inter-individual variability. Decisions made in the initial phases were correlated with the overall directions of the paths. Interestingly, the sound pressure levels of stimuli did not greatly influence several features of phonotactic paths such as sinuosity, walking bout lengths and durations. In order to ascertain the extent of our understanding of walking phonotaxis, a stochastic model was used to simulate the behaviour observed in the experiment. The model incorporated data from the experiment and our current understanding of field cricket auditory physiology. This model, based on stochastic turning towards the louder side, successfully recaptured several qualitative and quantitative features of the observed phonotactic paths. The simulation also reproduced the paths observed in a separate outdoor field experiment. Virtual crickets that were unilaterally deafened or had poor ear directionality exhibited walking paths similar to those observed in previous experiments. PMID- 17921168 TI - Linkage mechanics and power amplification of the mantis shrimp's strike. AB - Mantis shrimp (Stomatopoda) generate extremely rapid and forceful predatory strikes through a suite of structural modifications of their raptorial appendages. Here we examine the key morphological and kinematic components of the raptorial strike that amplify the power output of the underlying muscle contractions. Morphological analyses of joint mechanics are integrated with CT scans of mineralization patterns and kinematic analyses toward the goal of understanding the mechanical basis of linkage dynamics and strike performance. We test whether a four-bar linkage mechanism amplifies rotation in this system and find that the rotational amplification is approximately two times the input rotation, thereby amplifying the velocity and acceleration of the strike. The four-bar model is generally supported, although the observed kinematic transmission is lower than predicted by the four-bar model. The results of the morphological, kinematic and mechanical analyses suggest a multi-faceted mechanical system that integrates latches, linkages and lever arms and is powered by multiple sites of cuticular energy storage. Through reorganization of joint architecture and asymmetric distribution of mineralized cuticle, the mantis shrimp's raptorial appendage offers a remarkable example of how structural and mechanical modifications can yield power amplification sufficient to produce speeds and forces at the outer known limits of biological systems. PMID- 17921169 TI - Cross-modal effects on learning: a seismic stimulus improves color discrimination learning in a jumping spider. AB - The production of multimodal signals during animal displays is extremely common, and the function of such complex signaling has received much attention. Currently, the most frequently explored hypotheses regarding the evolution and function of complex signaling focus on the signal and/or signaler, or the signaling environment, while much less attention has been placed on the receivers. However, recent studies using vertebrates suggest that receiver psychology (e.g. learning and memory) may play a large role in the evolution of complex signaling. To date, the influence of multimodal cues on receiver learning and/or memory has not been studied in invertebrates. Here, we test the hypothesis that the presence of a seismic (vibratory) stimulus improves color discrimination learning in the jumping spider Habronattus dossenus. Using a heat-aversion learning experiment, we found evidence for a cross-modal effect on color learning. Over a series of training trials, individuals exposed to a seismic stimulus jumped onto the heated color less frequently and remained there for less time than did individuals not exposed to a seismic stimulus. In addition, in a final no-heat test trial, individuals from the seismic-present treatment were more likely to avoid the previously heated color than were individuals from the seismic-absent treatment. This is the first study to demonstrate a cross-modal influence on learning in an invertebrate. PMID- 17921170 TI - XtalPred: a web server for prediction of protein crystallizability. AB - XtalPred is a web server for prediction of protein crystallizability. The prediction is made by comparing several features of the protein with distributions of these features in TargetDB and combining the results into an overall probability of crystallization. XtalPred provides: (1) a detailed comparison of the protein's features to the corresponding distribution from TargetDB; (2) a summary of protein features and predictions that indicate problems that are likely to be encountered during protein crystallization; (3) prediction of ligands; and (4) (optional) lists of close homologs from complete microbial genomes that are more likely to crystallize. AVAILABILITY: The XtalPred web server is freely available for academic users on http://ffas.burnham.org/XtalPred PMID- 17921171 TI - Annotation of selection strengths in viral genomes. AB - MOTIVATION: Viral genomes tend to code in overlapping reading frames to maximize informational content. This may result in atypical codon bias and particular evolutionary constraints. Due to the fast mutation rate of viruses, there is additional strong evidence for varying selection between intra- and intergenomic regions. The presence of multiple coding regions complicates the concept of K(a)/K(s) ratio, and thus begs for an alternative approach when investigating selection strengths. Building on the paper by McCauley and Hein, we develop a method for annotating a viral genome coding in overlapping reading frames. We introduce an evolutionary model capable of accounting for varying levels of selection along the genome, and incorporate it into our prior single sequence HMM methodology, extending it now to a phylogenetic HMM. Given an alignment of several homologous viruses to a reference sequence, we may thus achieve an annotation both of coding regions as well as selection strengths, allowing us to investigate different selection patterns and hypotheses. RESULTS: We illustrate our method by applying it to a multiple alignment of four HIV2 sequences, as well as of three Hepatitis B sequences. We obtain an annotation of the coding regions, as well as a posterior probability for each site of the strength of selection acting on it. From this we may deduce the average posterior selection acting on the different genes. Whilst we are encouraged to see in HIV2, that the known to be conserved genes gag and pol are indeed annotated as such, we also discover several sites of less stringent negative selection within the env gene. To the best of our knowledge, we are the first to subsequently provide a full selection annotation of the Hepatitis B genome by explicitly modelling the evolution within overlapping reading frames, and not relying on simple K(a)/K(s) ratios. PMID- 17921172 TI - I/NI-calls for the exclusion of non-informative genes: a highly effective filtering tool for microarray data. AB - MOTIVATION: DNA microarray technology typically generates many measurements of which only a relatively small subset is informative for the interpretation of the experiment. To avoid false positive results, it is therefore critical to select the informative genes from the large noisy data before the actual analysis. Most currently available filtering techniques are supervised and therefore suffer from a potential risk of overfitting. The unsupervised filtering techniques, on the other hand, are either not very efficient or too stringent as they may mix up signal with noise. We propose to use the multiple probes measuring the same target mRNA as repeated measures to quantify the signal-to-noise ratio of that specific probe set. A Bayesian factor analysis with specifically chosen prior settings, which models this probe level information, is providing an objective feature filtering technique, named informative/non-informative calls (I/NI calls). RESULTS: Based on 30 real-life data sets (including various human, rat, mice and Arabidopsis studies) and a spiked-in data set, it is shown that I/NI calls is highly effective, with exclusion rates ranging from 70% to 99%. Consequently, it offers a critical solution to the curse of high-dimensionality in the analysis of microarray data. AVAILABILITY: This filtering approach is publicly available as a function implemented in the R package FARMS (www.bioinf.jku.at/software/farms/farms.html). PMID- 17921173 TI - Identification and visualization of cage-shaped proteins. AB - Cage-shaped protein, with its special structure, may have potential applications in biomedicine and nanotechnology. We developed a program CSPro (Cage-Shaped Protein) for efficient identification of cage-shaped proteins based on quaternary structure. CSPro is capable of revealing the corresponding cage-shaped feature more clearly and quickly than traditional visualization tools. Using CSPro, we have searched the full set of PDB (protein data bank) and three types of proteins are retrieved with notably large central cavities inside. CSPro can be used to validate if the quaternary structure of a protein is cage shaped in molecular simulation. AVAILABILITY: http://www.cad.zju.edu.cn/home/humin PMID- 17921174 TI - OSCAR: one-class SVM for accurate recognition of cis-elements. AB - MOTIVATION: Traditional methods to identify potential binding sites of known transcription factors still suffer from large number of false predictions. They mostly use sequence information in a position-specific manner and neglect other types of information hidden in the proximal promoter regions. Recent biological and computational researches, however, suggest that there exist not only locational preferences of binding, but also correlations between transcription factors. RESULTS: In this article, we propose a novel approach, OSCAR, which utilizes one-class SVM algorithms, and incorporates multiple factors to aid the recognition of transcription factor binding sites. Using both synthetic and real data, we find that our method outperforms existing algorithms, especially in the high sensitivity region. The performance of our method can be further improved by taking into account locational preference of binding events. By testing on experimentally-verified binding sites of GATA and HNF transcription factor families, we show that our algorithm can infer the true co-occurring motif pairs accurately, and by considering the co-occurrences of correlated motifs, we not only filter out false predictions, but also increase the sensitivity. AVAILABILITY: An online server based on OSCAR is available at http://bioinfo.au.tsinghua.edu.cn/oscar. PMID- 17921175 TI - Guidelines for the diagnosis and treatment of non-ST segment elevation acute coronary syndromes. PMID- 17921176 TI - Structural analysis of multiprotein complexes by cross-linking, mass spectrometry, and database searching. AB - Most protein complexes are inaccessible to high resolution structural analysis. We report the results of a combined approach of cross-linking, mass spectrometry, and bioinformatics to two human complexes containing large coiled-coil segments, the NDEL1 homodimer and the NDC80 heterotetramer. An important limitation of the cross-linking approach, so far, was the identification of cross-linked peptides from fragmentation spectra. Our novel approach overcomes the data analysis bottleneck of cross-linking and mass spectrometry. We constructed a purpose-built database to match spectra with cross-linked peptides, define a score that expresses the quality of our identification, and estimate false positive rates. We show that our analysis sheds light on critical structural parameters such as the directionality of the homodimeric coiled coil of NDEL1, the register of the heterodimeric coiled coils of the NDC80 complex, and the organization of a tetramerization region in the NDC80 complex. Our approach is especially useful to address complexes that are difficult in addressing by standard structural methods. PMID- 17921177 TI - Proteomics analysis of cytokine-induced dysfunction and death in insulin producing INS-1E cells: new insights into the pathways involved. AB - Cytokines released by islet-infiltrating immune cells play a crucial role in beta cell dysfunction and apoptotic cell death in the pathogenesis of type 1 diabetes and after islet transplantation. RNA studies revealed complex pathways of genes being activated or suppressed during this beta-cell attack. The aim of the present study was to analyze protein changes in insulin-producing INS-1E cells exposed to inflammatory cytokines in vitro using two-dimensional DIGE. Within two different pH ranges we observed 2214 +/- 164 (pH 4-7) and 1641 +/- 73 (pH 6-9) spots. Analysis at three different time points (1, 4, and 24 h of cytokine exposure) revealed that the major changes were taking place only after 24 h. At this time point 158 proteins were altered in expression (4.1%, n = 4, p < or = 0.01) by a combination of interleukin-1beta and interferon-gamma, whereas only 42 and 23 proteins were altered by either of the cytokines alone, giving rise to 199 distinct differentially expressed spots. Identification of 141 of these by MALDI TOF/TOF revealed proteins playing a role in insulin secretion, cytoskeleton organization, and protein and RNA metabolism as well as proteins associated with endoplasmic reticulum and oxidative stress/defense. We investigated the interactions of these proteins and discovered a significant interaction network (p < 1.27e-05) containing 42 of the identified proteins. This network analysis suggests that proteins of different pathways act coordinately in a beta-cell dysfunction/apoptotic beta-cell death interactome. In addition the data suggest a central role for chaperones and proteins playing a role in RNA metabolism. As many of these identified proteins are regulated at the protein level or undergo post-translational modifications, a proteomics approach, as performed in this study, is required to provide adequate insight into the mechanisms leading to beta-cell dysfunction and apoptosis. The present findings may open new avenues for the understanding and prevention of beta-cell loss in type 1 diabetes. PMID- 17921178 TI - High purity amphotericin B. AB - OBJECTIVES: Amphotericin B (AmB) is a drug of choice for treatment of disseminated fungal infections, but its use is often associated with severe adverse effects. Our observation that generic formulations of AmB contain multiple polyene components led us to propose that removal of other polyenes would yield a high purity AmB (AmBHP) with an improved therapeutic index. METHODS: To test that premise, AmBHP was first isolated from generic AmB by semi preparative reverse phase high-pressure liquid chromatography and then its effects were compared in vitro and in vivo with those of commercial AmB formulations. RESULTS: AmBHP proved to be as active as generic AmB against Candida albicans in vitro and as efficacious as both generic and lipid-complexed AmB in a Candida-infected mouse model. AmBHP appeared to be less toxic to human THP-1 monocytic cells than was generic AmB at low concentrations (<2 microM), as indicated by exclusion of Trypan Blue and incorporation of [(3)H]thymidine. At higher concentrations, effects of AmBHP and generic AmB (Pharma-Tek AmB, PTAmB) on thymidine incorporation and cytosolic calcium concentration were similar. General toxicity to AmBHP in vivo, as indicated by its apparent LD(50) and survival of Candida-infected mice, was roughly twofold less than that to generic or lipid-complexed AmB. Likewise, AmBHP decreased mean glomerular filtration rate about half as much as did a 10-fold lower dose of PTAmB. CONCLUSIONS: Taken together, these data indicate that AmBHP may represent a refinement of currently marketed AmB formulations, offering equal, if not better, efficacy with less toxicity. PMID- 17921179 TI - Recessive Twinkle mutations in early onset encephalopathy with mtDNA depletion. AB - Twinkle is a mitochondrial replicative helicase, the mutations of which have been associated with autosomal dominant progressive external ophthalmoplegia (adPEO), and recessively inherited infantile onset spinocerebellar ataxia (IOSCA). We report here a new phenotype in two siblings with compound heterozygous Twinkle mutations (A318T and Y508C), characterized by severe early onset encephalopathy and signs of liver involvement. The clinical manifestations included hypotonia, athetosis, sensory neuropathy, ataxia, hearing deficit, ophthalmoplegia, intractable epilepsy and elevation of serum transaminases. The liver showed mtDNA depletion, whereas the muscle mtDNA was only slightly affected. Alpers Huttenlocher syndrome has previously been associated with mutations of polymerase gamma, a replicative polymerase of mtDNA. We show here that recessive mutations of the close functional partner of the polymerase, the Twinkle helicase, can also manifest as early encephalopathy with liver involvement, a phenotype reminiscent of Alpers syndrome, and are a new genetic cause underlying tissue-specific mtDNA depletion. PMID- 17921180 TI - A new familial disease of saccadic oscillations and limb tremor provides clues to mechanisms of common tremor disorders. AB - Tremor disorders pose fundamental questions about disease mechanisms, and challenges to successful neurotherapeutics: What causes motor circuits to oscillate in disorders in which the central nervous system otherwise seems normal? How does inheritance 'determine' the clinical phenotype in familial tremor disorders? Here, we address these questions. Analogies between the neural circuits controlling rapid eye movements (saccades) and those controlling limb movements allow us to translate the interpretations from the saccadic systems to the limb movement system. Moreover, the relatively well understood neurophysiology of the ocular motor system offers a unique opportunity to test specific hypotheses about normal and abnormal motor control of both eye and limb movements. We describe a new familial disorder--'micro-saccadic oscillations and limb tremor (microSOLT)'--in a mother and daughter who had tiny saccadic oscillations of the eyes and tremor of the hands. This unique oscillatory movement disorder resembles other common tremor disorders (such as essential tremor) that occur in patients who have an otherwise normally functioning central nervous system. We hypothesize that microSOLT is caused by an inherited abnormality that results in abnormal membrane properties causing reduced external inhibition in the premotor neurons that generate the high-frequency discharge (burst) for saccades and for ballistic limb movements. To test this hypothesis, we recorded hand tremor and eye movements in two patients with microSOLT and particularly during natural circumstances when inhibition of the premotor saccadic burst neurons is removed (e.g. eye closure). We then simulated a conductance-based model for the premotor commands which included excitatory and reciprocally inhibitory burst neurons. The structure of this physiologically realistic model was based upon known cell types and anatomical connections in the brainstem (for saccades) and the thalamus (for limb movements). The physiological phenomenon of post-inhibitory rebound in premotor burst neurons makes the circuit inherently unstable and prone to oscillate unless prevented by external inhibition. Indeed, with simulated reduction of external inhibition (in this case glycinergic), saccadic oscillations and limb tremor were reproduced. Our results suggest that a single-inherited deficit can alter membrane properties, which impairs inhibition in an inherently unstable neural circuit causing the eye and limb oscillations in microSOLT. This concept has broad implications for understanding the mechanism and designing rationale pharmacotherapy for abnormal oscillations and may be applicable to other common disorders in which there are no structural abnormalities in the brain such as essential tremor. PMID- 17921181 TI - Auditory processing disorder in children with reading disabilities: effect of audiovisual training. AB - Reading disability is associated with phonological problems which might originate in auditory processing disorders. The aim of the present study was 2-fold: first, the perceptual skills of average-reading children and children with dyslexia were compared in a categorical perception task assessing the processing of a phonemic contrast based on voice onset time (VOT). The medial olivocochlear (MOC) system, an inhibitory pathway functioning under central control, was also explored. Secondly, we investigated whether audiovisual training focusing on voicing contrast could modify VOT sensitivity and, in parallel, induce MOC system plasticity. The results showed an altered voicing sensitivity in some children with dyslexia, and that the most severely impaired children presented the most severe reading difficulties. These deficits in VOT perception were sometimes accompanied by MOC function abnormalities, in particular a reduction in or even absence of the asymmetry in favour of the right ear found in average-reading children. Audiovisual training significantly improved reading and shifted the categorical perception curve of certain children with dyslexia towards the average-reading children's pattern of voicing sensitivity. Likewise, in certain children MOC functioning showed increased asymmetry in favour of the right ear following audiovisual training. The training-related improvements in reading score were greatest in children presenting the greatest changes in MOC lateralization. Taken together, these results confirm the notion that some auditory system processing mechanisms are impaired in children with dyslexia and that audiovisual training can diminish these deficits. PMID- 17921182 TI - Inflammasomes and rheumatic diseases: evolving concepts. AB - The realisation that the production of inflammatory cytokines in inflammatory rheumatic diseases may be induced by non-infectious endogenous signals has encouraged researchers to explore mechanisms of innate immunity and their contribution to the pathogenesis of these diseases. The nucleotide-binding and oligomerisation domain (NOD)-like receptors (NLRs) sense pathogens, products of damaged cells or endogenous metabolites and could potentially be involved in the initiation, amplification and progression of the inflammatory response in rheumatic diseases. NLRs are involved in the regulation of innate immune responses with some of them promoting the activation of inflammatory caspases within multiprotein complexes, called inflammasomes. A typical inflammasome consists of a sensor, an NLR protein, an adaptor protein such as ASC (for apoptosis-associated speck-like protein containing a caspase recruitment domain (CARD)) and an effector protein that is a caspase that activates pro-inflammatory cytokines such as interleukin (IL)1beta and IL18. Recent data suggest a role of the inflammasome in the pathogenesis of autoinflammatory as well as inflammatory rheumatic diseases such as juvenile chronic arthritis, adult onset Still disease, rheumatoid arthritis and gout. Modulation of these pathways may be a potential therapeutic target for inflammatory rheumatic diseases. PMID- 17921183 TI - Recommendations for the content and conduct of European League Against Rheumatism (EULAR) musculoskeletal ultrasound courses. AB - OBJECTIVE: To develop education guidelines for the conduct of future European League Against Rheumatism (EULAR) musculoskeletal ultrasound (MSUS) courses. METHODS: We undertook a consensus-based, iterative process using two consecutive questionnaires sent to 29 senior ultrasonographer rheumatologists who comprised the faculty of the 14th EULAR ultrasound course (June 2007). The first questionnaire encompassed the following issues: type of MSUS educational model; course timing; course curriculum; course duration; number of participants per teacher in practical sessions; time spent on hands-on sessions; and the requirements and/or restrictions for attendance at the courses. The second questionnaire consisted of questions related to areas where consensus had not been achieved in the first questionnaire, and to the topics and pathologies to be assigned to different educational levels. RESULTS: The response rate was 82.7% from the first questionnaire and 87.5% from the second questionnaire. The respondents were from 11 European countries. The group consensus on guidelines and curriculum was for a three-level education model (basic, intermediate and advanced) with timing and location related to the annual EULAR Congresses. The topics and pathologies to be included in each course were agreed. The course duration will be 20 h. There will be a maximum of six participants per teacher and 50-60% of total time will be spent on practical sessions. There was also agreement on prerequisite experience before attending the intermediate and advanced courses. CONCLUSION: We have developed European agreed guidelines for the content and conduct of EULAR ultrasound courses, which may also be recommended to national and local MSUS training programmes. PMID- 17921184 TI - Functional, molecular and proteomic characterisation of bone marrow mesenchymal stem cells in rheumatoid arthritis. AB - OBJECTIVE: Bone marrow (BM) mesenchymal stem cells (MSCs) are being considered as potential therapeutic agents in various inflammatory autoimmune diseases for their tissue-repair and anti-inflammatory tissue-protective properties. This study investigates the reserves and function, the molecular and proteomic profile and the differentiation potential of BM MSCs in patients with active rheumatoid arthritis (RA). METHODS: We evaluated the frequency of MSCs in the BM mononuclear cell fraction using a limiting dilution assay, the proliferative/clonogenic potential and the capacity of cells to differentiate towards the osteogenic/chondrogenic/adipogenic lineages using appropriate culture conditions. We also assessed the molecular and proteomic characteristics in terms of inflammatory cytokine gene and protein expression, the relative telomere length and the survival characteristics of BM MSCs. RESULTS: MSCs from patients with RA (n = 26) and age- and sex-matched healthy individuals (n = 21) were similar in frequency, differentiation potential, survival, immunophenotypic characteristics, and protein profile. Patient MSCs, however, had impaired clonogenic and proliferative potential in association with premature telomere length loss. Transcriptome analysis revealed differential expression of genes related to cell adhesion processes and cell cycle progression beyond the G1 phase. Previous treatment with methotrexate, corticosteroids, anti-cytokine and biological agents or other disease-modifying anti-inflammatory drugs did not correlate with the clonogenic and proliferative impairment of BM MSCs. CONCLUSION: In spite of some restrictions related to the impaired clonogenic and proliferative potential, our findings support the use of autologous BM MSCs in RA and may have important implications for the ongoing efforts to repair tissue injury commonly seen in the course of the disease. PMID- 17921185 TI - Efficacy and safety of the selective co-stimulation modulator abatacept following 2 years of treatment in patients with rheumatoid arthritis and an inadequate response to anti-tumour necrosis factor therapy. AB - OBJECTIVE: To evaluate the safety and efficacy of abatacept during 2 years of the ATTAIN (Abatacept Trial in Treatment of Anti-TNF INadequate responders) trial in patients with rheumatoid arthritis. METHODS: Patients completing the 6-month, double-blind period were eligible to enter the long-term extension; patients received abatacept approximately 10 mg/kg, plus disease-modifying antirheumatic drugs. Safety and efficacy (American College of Rheumatology (ACR) criteria responses, DAS28 (C-reactive protein), HAQ-DI, SF-36, Medical Outcomes Study Sleep Problems Index, fatigue VAS) were assessed through 2 years. RESULTS: 317 patients (218 from the abatacept and 99 from the placebo group) entered and 222 (70%) completed 18 months of long-term extension treatment. The incidence and type of adverse events were consistent between the double-blind and cumulative (double-blind plus long-term extension) periods. Rates of serious adverse events were 25.6 and 23.4 per 100 patient-years in the double-blind versus cumulative period. At 6 months and 2 years, using non-responder analyses, ACR responses in abatacept-treated patients were: ACR 20, 59.4% and 56.2%; ACR 50, 23.5% and 33.2%; ACR 70, 11.5% and 16.1%; HAQ-DI responses were 54.4% and 47.9%. At 6 months and 2 years, using post-hoc as-observed analyses, the percentage of patients (95% confidence interval) achieving DAS28 (C-reactive protein) low disease activity score (< or = 3.2) and DAS28 (C-reactive protein)-defined remission (< 2.6) increased from 18.3% (13.0, 23.5) to 32.0% (24.6, 39.4) and 11.1% (6.8, 15.3) to 20.3% (13.9, 26.6). Clinically meaningful improvements in SF 36, pain, fatigue and sleep problems were also maintained throughout the 2 years of abatacept treatment. CONCLUSION: No unique safety observations were reported during open-label exposure. Improvements in the signs and symptoms of rheumatoid arthritis, physical function and health-related quality of life observed after 6 months, were maintained throughout the 2 years in this population with difficult to-treat disease. TRIAL REGISTRATION NUMBER: NCT00124982. PMID- 17921186 TI - Distinct mechanisms of ethanol potentiation of local and paracapsular GABAergic synapses in the rat basolateral amygdala. AB - Converging lines of behavioral and pharmacological evidence suggest that GABAergic synapses in the basolateral amygdala (BLA) may play an integral role in mediating the anxiolytic effects of ethanol (EtOH). Since anxiety is thought to play an important role in the development of, and relapse to, alcoholism, elucidating the mechanisms through which EtOH modulates GABAergic synaptic transmission in the BLA may be fundamental in understanding the etiology of this disease. A recent study in mice has shown that principal cells within the BLA receive inhibitory input from two distinct types of GABAergic interneurons: a loosely distributed population of local interneurons and a dense network of paracapsular (pcs) GABAergic cells clustered along the external capsule border. Here, we sought to confirm the presence of these two populations of GABAergic synapses in the rat BLA and evaluate their ethanol sensitivity. Our results suggest that rat BLA pyramidal cells receive distinct inhibitory input from local and pcs interneurons and that EtOH potentiates both populations of synapses, albeit via distinct mechanisms. EtOH enhancement of local inhibitory postsynaptic currents (IPSCs) was associated with a significant decrease in paired-pulse ratio (PPR) and was significantly potentiated by the GABA(B) receptor antagonist SCH 50911 [(+)-(S)-5,5-dimethylmorpholinyl-2-acetic acid], consistent with a facilitation of GABA release from presynaptic terminals. Conversely, EtOH enhancement of pcs IPSCs did not alter PPR and was not enhanced by SCH 50911 but was inhibited by blockade of noradrenergic receptors. Collectively, these data reveal that EtOH can potentiate GABAergic inhibitory synaptic transmission in the rat BLA through at least two distinct pathways. PMID- 17921188 TI - Intrathecal administration of proteinase-activated receptor-2 agonists produces hyperalgesia by exciting the cell bodies of primary sensory neurons. AB - Proteinase-activated receptors (PARs) are a family of G-protein-coupled receptors that are activated by endogenous serine proteinases that cleave the N-terminal domain of the receptor unmasking a "tethered ligand" sequence. Trypsin and other agonists at PAR(2) act on peripheral nerves to augment the transfer of nociceptive information. We tested whether PAR(2) agonists also exert a spinal pronociceptive effect by i.t. administering the selective ligand, Ser-Leu-Ile-Gly Arg-Leu-NH(2) (SLI-GRL). This produced thermal and mechanical hyperalgesia in rats and mice and augmented mechanical and thermal hyperalgesia seen in the formalin inflammatory pain test. Effects of SLIGRL were abrogated in PAR(2) deficient mice and were not seen with the inactive control peptide, Leu-Arg-Gly Ile-Leu-Ser-NH(2). Surprisingly, electrophysiological studies, using whole-cell recording from rat substantia gelatinosa neurons, failed to demonstrate an increase in excitatory transmission or neuronal excitability following treatment with SLIGRL or trypsin. In fact, the actions of trypsin were consistent with a decrease in dorsal horn excitability. SLIGRL and trypsin did, however, depolarize and increase the excitability of large, medium and small primary afferent, dorsal root ganglion neurons. The effects were associated with an increase in conductance at hyperpolarized potentials and a decrease in conductance at depolarized potentials. PAR(2)-like immunoreactivity was found in DRG but not in spinal dorsal horn. These results suggest that activation of DRG neuron cell bodies may account for the pronociceptive actions of i.t. applied PAR(2) agonists. They also imply that pathophysiological release of PAR(2)-activating proteases in the vicinity of DRG neurons may produce profound effects on nociceptive processing in vivo. PMID- 17921187 TI - Glucuronidation of monohydroxylated warfarin metabolites by human liver microsomes and human recombinant UDP-glucuronosyltransferases. AB - Our understanding of human phase II metabolic pathways which facilitate detoxification and excretion of warfarin (Coumadin) is limited. The goal of this study was to test the hypothesis that there are specific human hepatic and extrahepatic UDP-glucuronosyltransferase (UGT) isozymes, which are responsible for conjugating warfarin and hydroxylated metabolites of warfarin. Glucuronidation activity of human liver microsomes (HLMs) and eight human recombinant UGTs toward (R)- and (S)-warfarin, racemic warfarin, and major cytochrome P450 metabolites of warfarin (4'-, 6-, 7-, 8-, and 10-hydroxywarfarin) has been assessed. HLMs, UGT1A1, 1A8, 1A9, and 1A10 showed glucuronidation activity toward 4'-, 6-, 7-, and/or 8-hydroxywarfarin with K(m) values ranging from 59 to 480 microM and V(max) values ranging from 0.03 to 0.78 microM/min/mg protein. Tandem mass spectrometry studies and structure comparisons suggested glucuronidation was occurring at the C4'-, C6-, C7-, and C8-positions. Of the hepatic UGT isozymes tested, UGT1A9 exclusively metabolized 8-hydroxywarfarin, whereas UGT1A1 metabolized 6-, 7-, and 8-hydroxywarfarin. Studies with extrahepatic UGT isoforms showed that UGT1A8 metabolized 7- and 8-hydroxywarfarin and that UGT1A10 glucuronidated 4'-, 6-, 7-, and 8-hydroxywarfarin. UGT1A4, 1A6, 1A7, and 2B7 did not have activity with any substrate, and none of the UGT isozymes evaluated catalyzed reactions with (R)- and (S)-warfarin, racemic warfarin, or 10-hydroxywarfarin. This is the first study identifying and characterizing specific human UGT isozymes, which glucuronidate major cytochrome P450 metabolites of warfarin with similar metabolic rates known to be associated with warfarin metabolism. Continued characterization of these pathways may enhance our ability to reduce life-threatening and costly complications associated with warfarin therapy. PMID- 17921189 TI - Different mitogen-activated protein kinase-dependent cytokine responses in cells of the monocyte lineage. AB - Macrophages release cytokines that may contribute to the chronic inflammation observed in pulmonary conditions such as asthma and chronic obstructive pulmonary disease. Thus, inhibition of macrophage cytokine production may have a therapeutic benefit. Human lung macrophages are a rich source of the proinflammatory cytokines, tumor necrosis factor (TNF)-alpha, granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-6, and IL-8, that are elevated in the bronchoalveolar lavage and sputum of subjects with respiratory diseases. Cytokine production from both monocytes and macrophages is mediated by mitogen-activated protein kinase (MAPK) pathways. This study compared the effects of a novel p38 MAPK inhibitor, N-cyano-N'-(2-{[8-(2,6-difluorophenyl) 4-(4-fluoro-2-methylphenyl)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2 yl]amino}ethyl)-guanidine (PCG), and an extracellular signal-regulated kinase (ERK) pathway inhibitor, 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one (PD098059), on cytokine release from lipopolysaccharide (LPS)-stimulated human monocytes, monocyte-derived macrophages (MDM), and lung macrophages. Lung macrophages, MDM, and monocytes were stimulated with LPS, and cytokine release was measured by enzyme-linked immunosorbent assay. Immunoblots were performed to confirm p38 and ERK1/2 MAPK expression and activity. PCG inhibited TNF-alpha release more effectively from monocytes compared with MDM or macrophages (maximal inhibition was 99.3 +/- 1.4, 62.7 +/- 4.3, and 58.6 +/- 6.6%, respectively; n = 7 9). PD098059 was less effective at suppressing TNF-alpha release from monocytes compared with MDM and lung macrophages (maximal inhibition was 37.4 +/- 2.8, 70.1 +/- 4.5, and 68.7 +/- 5.1%, respectively; n = 7-9). The pattern of GM-CSF, IL-6, and IL-8 release was comparable with that of TNF-alpha. These data suggest a differential involvement for each of these MAPK pathways in macrophage cytokine production compared with monocytes. PMID- 17921190 TI - Demonstration of proof of mechanism and pharmacokinetics and pharmacodynamic relationship with 4'-cyano-biphenyl-4-sulfonic acid (6-amino-pyridin-2-yl)-amide (PF-915275), an inhibitor of 11 -hydroxysteroid dehydrogenase type 1, in cynomolgus monkeys. AB - Glucocorticoids, through activation of the glucocorticoid receptor (GR), regulate hepatic gluconeogenesis. Elevated hepatic expression and activity of 11beta hydroxysteroid dehydrogenase type 1 (11betaHSD1) play a key role in ligand induced activation of the GR through the production of cortisol. Evidence from genetically modified mice suggests that inhibition of 11betaHSD1 might be a therapeutic approach to treat the metabolic syndrome. We have identified a potent 11betaHSD1 inhibitor, 4'-cyano-biphenyl-4-sulfonic acid (6-amino-pyridin-2-yl) amide (PF-915275), that is selective for the primate and human enzymes. The objective of this study was to demonstrate target inhibition with PF-915275 and to quantify the relationship between target inhibition and drug exposure in monkeys. We characterized the ability of PF-915275 to inhibit the conversion of prednisone, a synthetic cortisone analog that can be distinguished from the endogenous substrate cortisone, enabling a direct measure of substrate to product conversion without the complication of feedback. Adult cynomolgus monkeys were administered either vehicle or various doses of PF-915275 followed by a 10-mg/kg dose of prednisone. Prednisone conversion to prednisolone and the concentrations of PF-915275 were measured by liquid chromatography/tandem mass spectrometry. PF 915275 dose-dependently inhibited 11betaHSD1-mediated conversion of prednisone to prednisolone, with a maximum of 87% inhibition at a 3-mg/kg dose. An exposure response relationship was demonstrated, with an estimated EC(50) of 391 nM (total) and 17 nM (free). Insulin levels were also reduced in a dose-related manner. These results should enable the development of a biomarker for evaluating target modulation in humans that will aid in identifying 11betaHSD1 inhibitors to treat diabetes and other related metabolic diseases. PMID- 17921192 TI - Commentary: Genetic association studies see light at the end of the tunnel. PMID- 17921191 TI - Carrier-mediated transport of the quaternary ammonium neuronal nicotinic receptor antagonist n,n'-dodecylbispicolinium dibromide at the blood-brain barrier. AB - The quaternary ammonium compound N,N'-dodecyl-bispicolinium dibromide (bPiDDB) potently and selectively inhibits nicotinic receptors (nAChRs) mediating nicotine evoked [(3)H]dopamine release and decreases nicotine self-administration, suggesting that this polar, charged molecule penetrates the blood-brain barrier (BBB). This report focuses on 1) BBB penetration of bPiDDB; 2) the mechanism of permeation; and 3) comparison of bPiDDB to the cations choline and N octylnicotinium iodide (NONI), both of which are polar, charged molecules that undergo facilitated BBB transport. The BBB permeation of [(3)H]choline, [(3)H]NONI, and [(14)C]bPiDDB was evaluated using in situ rat brain perfusion methods. Cerebrovascular permeability surface-area product (PS) values for [(3)H]choline, [(3)H]NONI, and [(14)C]bPiDDB were comparable (1.33 +/- 0.1, 1.64 +/- 0.15, and 1.3 +/- 0.3 ml/s/g, respectively). To ascertain whether penetration was saturable, unlabeled substrate was added to the perfusion fluid. Unlabeled choline (500 microM) reduced the PS of [(3)H]choline to 0.15 +/- 0.06 microl/s/g (p < 0.01). Likewise, unlabeled bPiDDB (500 microM) reduced the PS of [(14)C]bPiDDB to 0.046 +/- 0.005 microl/s/g (p < 0.01), whereas unlabeled NONI reduced the PS for [(3)H]NONI by approximately 50% to 0.73 +/- 0.31 microl/s/g. The PS of [(14)C]bPiDDB was reduced (p < 0.05) in the presence of 500 microM choline, indicating that the BBB choline transporter may be responsible for the transport of bPiDDB into brain. Saturable kinetic parameters for [(14)C]bPiDDB were similar to those for [(3)H]choline. The current results suggest that bPiDDB uses the BBB choline transporter for approximately 90% of its permeation into brain, and they demonstrate the carrier-mediated BBB penetration of a novel bisquaternary ammonium nAChR antagonist. PMID- 17921193 TI - Commentary: preventing colorectal cancer with aspirin what next? PMID- 17921194 TI - Commentary: aspirin and cancer prevention. PMID- 17921195 TI - Colorectal cancer risk, chronic illnesses, operations and medications: case control results from the Melbourne Colorectal Cancer Study. 1988. AB - The associations between colorectal cancer risk and several chronic illnesses, operations and various medications were examined in 715 colorectal cancer cases and 727 age- and sex-matched controls in data derived from a large, comprehensive population-based study of this cancer conducted in Melbourne, Australia. There was a statistically significant deficit among cases of hypertension, heart disease, stroke, chronic chest disease and chronic arthritis and a statistically significant excess of 'haemorrhoids' among cases, and all of these differences were consistent for both colon and rectal cancers and for both males and females. Although no statistically significant differences were found for other cancers, there were twice as many breast cancers among cases (16) than among controls (8) and also there were 9 uterine cancers among cases and only 2 among controls. There was a statistically significant deficit among cases in the use of aspirin containing medication and vitamin supplements and this was consistent for both colon and rectal cancers and for both males and females. There was a statistically significant excess of large bowel polypectomy among cases. The modelling of these significant associations simultaneously in a logistic regression equation indicated that hypertension, heart disease, chronic arthritis and aspirin use were each independent effects and consistent for both colon and rectal cancers for both males and females and also that these effects were independent of dietary risk factors previously described in the Melbourne study. The possible relevance of these findings towards an understanding of colorectal cancer risk and aetiology is discussed. PMID- 17921196 TI - Similarities, overlap and differences between burnout and prolonged fatigue in the working population. AB - BACKGROUND: Burnout and prolonged fatigue are related but distinct concepts that have seldom been empirically compared. AIM: To examine similarities, overlap and differences between burnout and prolonged fatigue. DESIGN: Observational study. METHODS: We analysed baseline data from the Maastricht Cohort Study on Fatigue at Work (n = 12 140). The discriminative abilities of the Checklist Individual Strength (CIS) and the Maslach Burnout Inventory-General Survey (MBI-GS) were evaluated using principal component analysis. Overlap, similarities and differences regarding health, work and demographic factors between subgroups were assessed. RESULTS: The discriminative abilities of the CIS and MBI-GS appeared to be moderate. Prolonged fatigue and burnout cases overlapped considerably. The subgroup consisting of cases with concurrent fatigue and burnout tended to have poorer outcomes in terms of health and work factors than the subgroups with either prolonged fatigue or burnout. Similar patterns were found for subjective fatigue and exhaustion. DISCUSSION: There appear to be some relevant differences between burnout and prolonged fatigue, with respect to work and health factors. Burnout and prolonged fatigue can occur both separately and simultaneously. Having both conditions simultaneously seems to be associated with worse outcomes than having either alone. PMID- 17921197 TI - Anaemia is common and predicts mortality in diabetic nephropathy. AB - BACKGROUND: Diabetes is the single largest cause of chronic renal failure, accounting for 18% of patients on renal replacement therapy in the UK. AIM: To investigate the chronic kidney disease stage at which patients with diabetic nephropathy are referred to renal services, determine the prevalence of anaemia in patients with diabetic nephropathy, examine patient outcome and identity prognostic factors. DESIGN: Retrospective review. METHODS: Patients with diabetic nephropathy referred to our renal services between 1989 and 2004 were identified from electronic records. Estimated glomerular filtration rate (calculated using the MDRD formula) and haemoglobin at referral were collected. Times to renal replacement therapy and death were noted. RESULTS: We identified 508 patients. At referral, mean eGFR was 34 ml/min/1.73 m(2) and 48% of patients were at CKD stages 4 and 5. Mean haemoglobin was 11.7 g/dl; 21% had a haemoglobin <10 g/dl at referral. Median survival was 37.9 months (95%CI 33.2-42.6); median survival independent of renal replacement therapy (RRT) was 21 months (95%CI 17.8-24.6). Of patients starting RRT, 38% did so within 1 year of referral. Older age (RR 1.02, 95%CI 1.01-1.04) and lower haemoglobin (RR 0.9, 95%CI 0.85-0.99) at referral predicted death on multivariate analysis. DISCUSSION: At referral to renal services, almost 50% of patients with diabetic nephropathy were at CKD stages 4 and 5. Anaemia was common and predicted mortality. All diabetic patients from CKD stage 3 should be screened for anaemia. We believe that patients with diabetic nephropathy should be discussed with renal services when they reach CKD stage 3 with evidence of progression of renal disease. PMID- 17921198 TI - Is there a role for low carbohydrate diets in the management of type 2 diabetes? PMID- 17921200 TI - Molecular mechanics of filamin's rod domain. AB - Rearrangement of the actin cytoskeleton is integral to cell shape and function. Actin-binding proteins, e.g., filamin, can naturally contribute to the mechanics and function of the actin cytoskeleton. The molecular mechanical bases for filamin's function in actin cytoskeletal reorganization are examined here using molecular dynamics simulations. Simulations are performed by applying forces ranging from 25 pN to 125 pN for 2.5 ns to the rod domain of filamin. Applying small loads ( approximately 25 pN) to filamin's rod domain supplies sufficient energy to alter the conformation of the N-terminal regions of the rod. These forces break local hydrogen bond coordination often enough to allow side chains to find new coordination partners, in turn leading to drastic changes in the conformation of filamin, for example, increasing the hydrophobic character of the N-terminal rod region and, alternatively, activating the C-terminal region to become increasingly stiff. These changes in conformation can lead to changes in the affinity of filamin for its binding partners. Therefore, filamin can function to transduce mechanical signals as well as preserve topology of the actin cytoskeleton throughout the rod domain. PMID- 17921201 TI - A quantitative model for the all-or-none permeabilization of phospholipid vesicles by the antimicrobial peptide cecropin A. AB - The mechanism of the all-or-none release of the contents of phospholipid vesicles induced by the antimicrobial peptide cecropin A was investigated. A detailed experimental study of the kinetics of dye release showed that the rate of release increases with the ratio of peptide bound per vesicle and, at constant concentration, with the fraction of the anionic lipid phosphatidylglycerol in neutral, phosphatidylcholine membranes. Direct measurement of the kinetics of peptide binding and dissociation from vesicles revealed that the on-rate is almost independent of vesicle composition, whereas the off-rate decreases by orders of magnitude with increasing content of anionic lipid. A simple, exact model fits all experimental kinetic data quantitatively. This is the first time that an exact kinetic model is implemented for a peptide with an all-or-none mechanism. In this model, cecropin A binds reversibly to vesicles, which at a certain point enter an unstable state. In this state, a pore probably opens and all vesicle contents are released, consistent with the all-or-none mechanism. This pore state is a state of the whole vesicle, but does not necessarily involve all peptides on that vesicle. No peptide oligomerization on the vesicle is involved; alternative models that assume oligomerization are inconsistent with the experimental data. Thus, formation of well-defined, peptide-lined pores is unlikely. PMID- 17921202 TI - Dynamics of the acetylcholinesterase tetramer. AB - Acetylcholinesterase rapidly hydrolyzes the neurotransmitter acetylcholine in cholinergic synapses, including the neuromuscular junction. The tetramer is the most important functional form of the enzyme. Two low-resolution crystal structures have been solved. One is compact with two of its four peripheral anionic sites (PAS) sterically blocked by complementary subunits. The other is a loose tetramer with all four subunits accessible to solvent. These structures lacked the C-terminal amphipathic t-peptide (WAT domain) that interacts with the proline-rich attachment domain (PRAD). A complete tetramer model (AChEt) was built based on the structure of the PRAD/WAT complex and the compact tetramer. Normal mode analysis suggested that AChEt could exist in several conformations with subunits fluctuating relative to one another. Here, a multiscale simulation involving all-atom molecular dynamics and C alpha-based coarse-grained Brownian dynamics simulations was carried out to investigate the large-scale intersubunit dynamics in AChEt. We sampled the ns-mus timescale motions and found that the tetramer indeed constitutes a dynamic assembly of monomers. The intersubunit fluctuation is correlated with the occlusion of the PAS. Such motions of the subunits "gate" ligand-protein association. The gates are open more than 80% of the time on average, which suggests a small reduction in ligand-protein binding. Despite the limitations in the starting model and approximations inherent in coarse graining, these results are consistent with experiments which suggest that binding of a substrate to the PAS is only somewhat hindered by the association of the subunits. PMID- 17921203 TI - An oxygen scavenging system for improvement of dye stability in single-molecule fluorescence experiments. AB - The application of single-molecule fluorescence techniques to complex biological systems places demands on the performance of single fluorophores. We present an enzymatic oxygen scavenging system for improved dye stability in single-molecule experiments. We compared the previously described protocatechuic acid/protocatechuate-3,4-dioxygenase system to the currently employed glucose oxidase/catalase system. Under standardized conditions, we observed lower dissolved oxygen concentrations with the protocatechuic acid/protocatechuate-3,4 dioxygenase system. Furthermore, we observed increased initial lifetimes of single Cy3, Cy5, and Alexa488 fluorophores. We further tested the effects of chemical additives in this system. We found that biological reducing agents increase both the frequency and duration of blinking events of Cy5, an effect that scales with reducing potential. We observed increased stability of Cy3 and Alexa488 in the presence of the antioxidants ascorbic acid and n-propyl gallate. This new O(2)-scavenging system should have wide application for single-molecule fluorescence experiments. PMID- 17921204 TI - Subcellular imaging of dynamic protein interactions by bioluminescence resonance energy transfer. AB - Despite the fact that numerous studies suggest the existence of receptor multiprotein complexes, visualization and monitoring of the dynamics of such protein assemblies remain a challenge. In this study, we established appropriate conditions to consider spatiotemporally resolved images of such protein assemblies using bioluminescence resonance energy transfer (BRET) in mammalian living cells. Using covalently linked Renilla luciferase and yellow fluorescent proteins, we depicted the time course of dynamic changes in the interaction between the V2-vasopressin receptor and beta-arrestin induced by a receptor agonist. The protein-protein interactions were resolved at the level of subcellular compartments (nucleus, plasma membrane, or endocytic vesicules) and in real time within tens-of-seconds to tens-of-minutes time frame. These studies provide a proof of principle as well as experimental parameters and controls required for high-resolution dynamic studies using BRET imaging in single cells. PMID- 17921205 TI - Electric field perturbations of spiral waves attached to millimeter-size obstacles. AB - Reentrant spiral waves can become pinned to small anatomical obstacles in the heart and lead to monomorphic ventricular tachycardia that can degenerate into polymorphic tachycardia and ventricular fibrillation. Electric field-induced secondary source stimulation can excite directly at the obstacle, and may provide a means to terminate the pinned wave or inhibit the transition to more complex arrhythmia. We used confluent monolayers of neonatal rat ventricular myocytes to investigate the use of low intensity electric field stimulation to perturb the spiral wave. A hole 2-4 mm in diameter was created in the center to pin the spiral wave. Monolayers were stained with voltage-sensitive dye di-4-ANEPPS and mapped at 253 sites. Spiral waves were initiated that attached to the hole (n = 10 monolayers). Electric field pulses 1-s in duration were delivered with increasing strength (0.5-5 V/cm) until the wave terminated after detaching from the hole. At subdetachment intensities, cycle length increased with field strength, was sustained for the duration of the pulse, and returned to its original value after termination of the pulse. Mechanistically, conduction velocity near the wave tip decreased with field strength in the region of depolarization at the obstacle. In summary, electric fields cause strength dependent slowing or detachment of pinned spiral waves. Our results suggest a means to decelerate tachycardia that may help to prevent wave degeneration. PMID- 17921206 TI - Hindered diffusion through an aqueous pore describes invariant dye selectivity of Cx43 junctions. AB - The permselectivity (permeance/conductance) of Cx43-comprised gap junctions is a variable parameter of junctional function. To ascertain whether this variability in junctional permselectivity is explained by heterogeneous charge or size selectivity of the comprising channels, the permeance of individual Cx43 gap junctions to combinations of two dyes differing in either size or charge was determined in four cell types: Rin43, NRKe, HeLa43, and cardiac myocytes. The results show that Cx43 junctions are size- but not charge-selective and that both selectivities are constant parameters of junctional function. The consistency of dye selectivities indicates that the large continuum of measured junctional permselectivities cannot be ascribed to an equivalent continuum of individual channel selectivities. Further, the relative dye permeance sequence of NBD-M-TMA approximately Alexa 350 > Lucifer yellow > Alexa 488 >> Alexa 594 (Stokes radii of 4.3 A, 4.4 A, 4.9 A, 5.8 A, and 7.4 A, respectively) and the conductance sequence of KCl > TEACl approximately Kglutamate are well described by hindered diffusion through an aqueous pore with radius approximately 10 A and length 160 A. The permselectivity and dye selectivity data suggest the variable presence in Cx43-comprised junctions of conductive channels that are either dye-impermeable or dye-permeable. PMID- 17921207 TI - Coarse-grained molecular dynamics simulations of phase transitions in mixed lipid systems containing LPA, DOPA, and DOPE lipids. AB - The mechanisms that mediate biomembrane shape transformations are of considerable interest in cell biology. Recent in vitro experiments show that the chemical transformation of minor membrane lipids can induce dramatic shape changes in biomembranes. Specifically, it was observed that the addition of DOPA to DOPE has no effect on the stability of the bilayer structure of the membrane. In contrast, the addition of LPA to DOPE stabilizes the bilayer phase of DOPE, increasing the temperature of a phase transition from the bilayer to the inverted hexagonal phase. This result suggests that the chemical conversion of DOPA to LPA is sufficient for triggering a dramatic change in the shape of biomembranes. The LPA/DOPA/DOPE mixture of lipids provides a simple model system for understanding the molecular events driving the shape change. In this work, we used coarse grained molecular dynamics simulations to study the phase transitions of this lipid mixture. We show that despite the simplicity of the coarse-grained model, it reproduces the experimentally observed phase changes of: 1), pure LPA and DOPA with respect to changes in the concentration of cations; and 2), LPA/DOPE and DOPA/DOPE mixtures with respect to temperature. The good agreement between the model and experiments suggests that the computationally inexpensive coarse grained approach can be used to infer macroscopic membrane properties. Furthermore, analysis of the shape of the lipid molecules demonstrates that the phase behavior of single-lipid systems is consistent with molecular packing theory. However, the phase stability of mixed lipid systems exhibits significant deviations from this theory, which suggests that the elastic energy of the lipids, neglected in the packing theory, plays an important role. PMID- 17921208 TI - The physical basis for the head-to-tail rule that excludes most fullerene cages from self-assembly. AB - In the companion article, we proposed that fullerene cages with head-to-tail dihedral angle discrepancies do not self-assemble. Here we show why. If an edge abuts a pentagon at one end and a hexagon at the other, the dihedral angle about the edge increases, producing a dihedral angle discrepancy (DAD) vector. The DADs about all five/six edges of a central pentagonal/hexagonal face are determined by the identities-pentagon or hexagon-of its five/six surrounding faces. Each "Ring" central face plus specific surrounding faces-may have zero, two, or four edges with DAD. In most Rings, the nonplanarity induced by DADs is shared among surrounding faces. However, in a Ring that has DADs arranged head of one to tail of another, the nonplanarity cannot be shared, so some surrounding faces would be especially nonplanar. Because the head-to-tail exclusion rule is an implicit geometric constraint, the rule may operate either by imposing a kinetic barrier that prevents assembly of certain Rings or by imposing an energy cost that makes those Rings unlikely to last in an equilibrium circumstance. Since Rings with head-to-tail DADs would be unlikely to self-assemble or last, fullerene cages with those Rings would be unlikely to self-assemble. PMID- 17921209 TI - A geometric principle may guide self-assembly of fullerene cages from clathrin triskelia and from carbon atoms. AB - Clathrin triskelia and carbon atoms alike self-assemble into a limited selection of fullerene cages (with n three connected vertices, 3n/2 edges, 12 pentagonal faces, and (n-20)/2 hexagonal faces). We show that a geometric constraint exclusion of head-to-tail dihedral angle discrepancies (DADs)-explains this limited selection as well as successful assembly into such closed cages in the first place. An edge running from a pentagon to a hexagon has a DAD, since the dihedral angles about the edge broaden from its pentagon (tail) end to its hexagon (head) end. Of the 21 configurations of a central face and surrounding faces, six have such DAD vectors arranged head-to-tail. Of the 5770 mathematically possible fullerene cages for n 50 Hz) with a slope of approximately -2.5. This type of frequency scaling cannot be accounted for by traditional models, as either single-compartment models or models based on reconstructed cell morphologies display a frequency scaling with a slope close to -4. This slope is due to the fact that the membrane resistance is short-circuited by the capacitance for high frequencies, a situation which may not be realistic. Here, we integrate nonideal capacitors in cable equations to reflect the fact that the capacitance cannot be charged instantaneously. We show that the resulting nonideal cable model can be solved analytically using Fourier transforms. Numerical simulations using a ball-and-stick model yield membrane potential activity with similar frequency scaling as in the experiments. We also discuss the consequences of using nonideal capacitors on other cellular properties such as the transmission of high frequencies, which is boosted in nonideal cables, or voltage attenuation in dendrites. These results suggest that cable equations based on nonideal capacitors should be used to capture the behavior of neuronal membranes at high frequencies. PMID- 17921221 TI - Decoding of calcium oscillations by phosphorylation cycles: analytic results. AB - Experimental studies have demonstrated that Ca(2+)-regulated proteins are sensitive to the frequency of Ca(2+) oscillations, and several mathematical models for specific proteins have provided insight into the mechanisms involved. Because of the large number of Ca(2+)-regulated proteins in signal transduction, metabolism and gene expression, it is desirable to establish in general terms which molecular properties shape the response to oscillatory Ca(2+) signals. Here we address this question by analyzing in detail a model of a prototypical Ca(2+) decoding module, consisting of a target protein whose activity is controlled by a Ca(2+)-activated kinase and the counteracting phosphatase. We show that this module can decode the frequency of Ca(2+) oscillations, at constant average Ca(2+) signal, provided that the Ca(2+) spikes are narrow and the oscillation frequency is sufficiently low--of the order of the phosphatase rate constant or below. Moreover, Ca(2+) oscillations activate the target more efficiently than a constant signal when Ca(2+) is bound cooperatively and with low affinity. Thus, the rate constants and the Ca(2+) affinities of the target-modifying enzymes can be tuned in such a way that the module responds optimally to Ca(2+) spikes of a certain amplitude and frequency. Frequency sensitivity is further enhanced when the limited duration of the external stimulus driving Ca(2+) signaling is accounted for. Thus, our study identifies molecular parameters that may be involved in establishing the specificity of cellular responses downstream of Ca(2+) oscillations. PMID- 17921222 TI - Characterization of synchronization in interacting groups of oscillators: application to seizures. AB - We investigate the emergence of synchronization in two groups of oscillators; one group acts as a synchronization source, and the other as the target. Based on phase model simulations, we construct a synchrony index (SI): a combination of intra- and intergroup synchronies. The SI characterizes the extent of induced synchrony in the population. We demonstrate the usefulness of the measure in a test case of mesial temporal lobe epilepsy: the SI can be readily calculated from standard electroencephalographic measurements. We show that the synchrony index has a statistically significant increased value for the ictal periods and that the epileptic focus can be located by identifying the most synchronous pairs of electrodes during the initial part of ictal period of the seizure. We also show that it is possible in this pilot case to differentiate clinical and subclinical seizures based on the dynamical features of the synchronization. The synchronization index was found to be a useful quantity for the characterization of "pathological hypersynchronization" within a well-characterized patient with mesial temporal lobe epilepsy and thus has potential medical value in seizure detection, localizing ability, and association with later surgical outcome. PMID- 17921223 TI - Analysis of FRET signals in the presence of free donors and acceptors. AB - A method for spectral analysis of Forster resonance energy transfer (FRET) signals is presented, taking into consideration both the contributions of unpaired donor and acceptor fluorophores and the influence of incomplete labeling of the interacting partners. It is shown that spectral analysis of intermolecular FRET cannot yield accurate values of the Forster energy transfer efficiency E, unless one of the interactors is in large excess and perfectly labeled. Instead, analysis of donor quenching yields a product of the form Ef(d)p(a), where f(d) is the fraction of donor-type molecules participating in donor-acceptor complexes and p(a) is the labeling probability of the acceptor. Similarly, analysis of sensitized emission yields a product involving Ef(a). The analysis of intramolecular FRET (e.g., of tandem constructs) yields the product Ep(a). We use our method to determine these values for a tandem construct of cyan fluorescent protein and yellow fluorescent protein and compare them with those obtained by standard acceptor photobleaching and fluorescence lifetime measurements. We call the method lux-FRET, since it relies on linear unmixing of spectral components. PMID- 17921224 TI - Munc-18-1 regulates the initial release rate of exocytosis. AB - Carbon fiber amperometry is a popular method for measuring single exocytotic events; however, the functional interpretation of the data can prove hazardous. For example, changes to vesicle transmitter levels can appear to cause changes in the timing and rate of the fusion process itself. Use of an analytical technique based on differentiation revealed that an increase in dense-core granule catecholamine content by exogenous application of l-DOPA did not affect initial release rates. Changes to the timing and amplitude of amperometric spikes from l DOPA-treated cells are, then, likely a reflection of the increased quantal size rather than any direct effect on exocytosis itself. Applying this new analysis to individual fusion events from cells expressing Munc-18-1 with various specific point mutations demonstrated that Munc-18-1 functions at a late stage involved in the determination of the initial rate of fusion. Furthermore, a mutation of the protein that inhibits its biochemical interaction with the t-SNARE syntaxin-1 in a closed conformation caused premature termination of the fusion event. Through these two late-stage functions, Munc-18-1 could act as a key protein involved in the presynaptic control of signaling strength and duration. PMID- 17921225 TI - Amplitude variability and extracellular low-pass filtering of neuronal spikes. AB - The influence of neural morphology and passive electrical parameters on the width and amplitude of extracellular spikes is investigated by combined analytical and numerical investigations of idealized and anatomically reconstructed pyramidal and stellate neuron models. The main results are: 1), All models yield a low-pass filtering effect, that is, a spike-width increase with increasing distance from soma. 2), A neuron's extracellular spike amplitude is seen to be approximately proportional to the sum of the dendritic cross-sectional areas of all dendritic branches connected to the soma. Thus, neurons with many, thick dendrites connected to soma will produce large amplitude spikes, and therefore have the largest radius of visibility. 3), The spike shape and amplitude are found to be dependent on the membrane capacitance and axial resistivity, but not on the membrane resistivity. 4), The spike-amplitude decay with distance r is found to depend on dendritic morphology, and is decaying as 1/r(n) with 1 or= 2 far away. PMID- 17921226 TI - Whole-brain N-acetylaspartate as a surrogate marker of neuronal damage in diffuse neurologic disorders. AB - Proton MR spectroscopy (1H-MR spectroscopy) is a quantitative MR imaging technique often used to complement the sensitivity of conventional MR imaging with specific metabolic information. A key metabolite is the amino acid derivative N-acetylaspartate (NAA), which is almost exclusive to neurons and their processes and is, therefore, an accepted marker of their health and attenuation. Unfortunately, most 1H-MR spectroscopy studies only account for small 1- to 200-cm volumes of interest (VOI), representing less than 20% of the total brain volume. These VOIs have at least 5 additional restrictions: 1) To avoid contamination from subcutaneous and bone marrow lipids, they must be placed away from the skull, thereby missing most of the cortex. 2) They must be image guided onto MR imaging-visible pathology, subjecting them to the implicit assumption that metabolic changes occur only there. 3) They encounter misregistration errors in serial studies. 4) The time needed to accumulate sufficient signal-intensity quality is often restrictive, and 5) they incur (unknown) T1- and T2-weighting. All these issues are avoided (at the cost of specific localization) by measuring the nonlocalized average NAA concentration over the entire brain. Indeed, whole-brain NAA quantification has been applied to several diffuse neurodegenerative diseases (where specific localization is less important than the total load of the pathology), and the results are presented in this review. PMID- 17921227 TI - Diffusion tensor imaging in patients with essential tremor. AB - BACKGROUND AND PURPOSE: The traditional paradigm has regarded essential tremor (ET) as a benign disorder. However, recent clinical, neuroimaging, and neuropathologic studies suggest that ET may be a progressive neurologic disorder. Based on clinicopathologic findings that cerebellum and its outflow are the key structures in ET and degeneration of gray matter in cerebellum is followed by consequent wallerian degeneration of white matter (WM) fibers, the aim of the present study was to investigate changes in anisotropy in patients with ET. MATERIALS AND METHODS: Fractional anisotropy (FA) images were generated from DTI data acquired at 1.5T in 10 patients with ET compared with 8 control subjects by using statistical parametric mapping to make voxel-by-voxel comparisons. RESULTS: Compared with the control subjects, the patients with ET exhibited significantly reduced FA (P(uncorrected) < .005) in the anterolateral portion of the right pons and decreased FA in the bilateral cerebellum, left retrorubral area of the midbrain, and bilateral deep WM, including the orbitofrontal, lateral frontal, parietal, and temporal WM. CONCLUSION: This study demonstrates that structural changes in the WM are extensive in patients with ET, supporting the findings of previous functional neuroimaging and pathologic studies. PMID- 17921228 TI - Differentiation of benign and malignant pathology in the head and neck using 3T apparent diffusion coefficient values: early experience. AB - BACKGROUND AND PURPOSE: The purpose of this work was to study differences in apparent diffusion coefficient (ADC) values between benign and malignant head and neck lesions at 3T field strength imaging. MATERIALS AND METHODS: Our study population in this retrospective study was derived from the patient population who had undergone routine neck 3T MR imaging (for clinical indications) from December 2005 to December 2006. There were 33 patients identified: 17 with benign and 16 with malignant pathologies. In all of the subjects, conventional MR imaging sequences were performed apart from diffusion-weighted sequences. The mean ADC values in the benign and malignant groups were compared using an unpaired t test with unequal variance with a P < 0.05 considered statistically significant. RESULTS: There was a statistically significant difference (P = .004) between the mean ADC values (in 10(-3) mm(2)/s) in the benign and malignant lesions (1.505 +/- 0.487; 95% confidence interval, 1.305-1.706, and 1.071 +/- 0.293; 95% confidence interval, 0.864-1.277, respectively). There were 2 malignant lesions with ADC values higher than 1.3 x 10(-3) mm(2)/s and 5 benign lesions with ADC values less than 1.3 x 10(-3) mm(2)/s. The lack of overlap of ADC values within 95% confidence limits suggests that a 3T ADC value of 1.3 x 10( 3) mm(2)/s may be the threshold value for differentiation between benign and malignant head and neck lesions. CONCLUSION: ADC values of benign and malignant neck pathologies are significantly different at 3T imaging, though larger studies are required to establish threshold ADC values that can applied in daily clinical practice. PMID- 17921229 TI - Endovascular treatment of intracranial aneurysms with matrix detachable coils: midterm anatomic follow-up from a prospective multicenter registry. AB - BACKGROUND AND PURPOSE: Polyglycolic/polylactic acid-covered platinum coils have been proposed to reduce the rate of aneurysm recanalization after endovascular treatment. A prospective and multicenter registry was conducted in France to evaluate the safety and short-term and midterm efficacy of Matrix coils. This analysis focused on anatomic midterm results. MATERIALS AND METHODS: Two hundred thirty-six patients harboring 244 ruptured or unruptured aneurysms treated via endovascular approach by using Matrix coils were included in this registry. Treatment was totally or partially performed by using Matrix coils. Anatomic results were evaluated on postoperative and last-follow-up digital subtraction angiography (DSA) by using the Raymond scale. "Recanalization" was defined as worsening, and "progressive thrombosis" was defined as improvement on the Raymond scale. RESULTS: Anatomic midterm follow-up was obtained in 165 of 236 patients (70%) harboring 171 aneurysms (range, 6-27 months; mean, 14 +/- 4 months). At midterm follow-up angiography, 79 aneurysms were completely occluded (46.2%), 43 had a neck remnant (25.1%), and 49 had an aneurysm remnant (28.7%). Of 171 aneurysms, recanalization was observed in 44 patients (25.7%), including major recanalization in 18 patients (10.5%). Recanalization was more frequent if the embolized volume of aneurysm was or =8-point National Institutes of Health Stroke Scale improvement at 24 hours occurred. Follow-up NCCT ASPECTS and 90-day modified Rankin score (mRS) were radiologic and clinical reference standards. Receiver operating characteristic curves derived optimal thresholds for outcome. RESULTS: Cerebral blood volume and NCCT ASPECTS had similar radiologic correlations (0.6 and 0.5, respectively) and best predicted infarct size in the absence of major neurologic improvement. A NCCT ASPECT threshold of 7 and a cerebral blood volume threshold of 8 discriminated patients with poor follow-up scans (P < .0002 and P = .0001) and mRS < or =2 (P = .001 and P < .001). Only cerebral blood volume predicted major neurologic improvement (P = .02). Interobserver agreement was substantial (intraclass correlation coefficient, 0.69). Cerebral blood volume ASPECTS sensitivity, specificity, positive predictive value, and negative predictive value for clinical outcome were 60%, 100%, 100%, and 45%, respectively. No patients with cerebral blood volume ASPECTS <8 achieved good clinical outcome. CONCLUSION: Cerebral blood volume ASPECTS is equivalent to NCCT for predicting radiologic outcome but may have an additional benefit in predicting patients with major neurologic improvement. PMID- 17921239 TI - subchronic inhalation toxicity study of 1,3-dichloro-2-propanol in rats. AB - The subchronic toxicity of 1,3-dichloro-2-propanol (1,3-DCP) was investigated in Fischer 344 rats after 13 weeks of repeated, whole-body inhalation exposure. Groups of 10 rats of each sex were exposed to 1,3-DCP vapor by whole-body inhalation exposure at concentrations of 0, 5, 20 or 80 ppm for 6 h/day, 5 days/week for 13 weeks. All of the rats were sacrificed at the end of the treatment period. During the test period, clinical signs, mortality, body weights, food consumption, ophthalmoscopy, urinalysis, hematology, serum biochemistry, gross findings, organ weights and histopathology were assessed. At 80 ppm, a decrease in the body weight gain, an increase in the urine protein and leukocyte counts and an increase in the liver and kidney weights were observed in both genders. Hematological and serum biochemical investigations revealed decreases in hemoglobin (HB), hematocrit (HCT), mean corpuscular volume (MCV) and mean corpuscular HB, as well as increases in the platelet (PLT) count, serum aspartate aminotransferase and alanine aminotransferase. The number of white blood cells was significantly lower in males than in controls, but this was not the case in females. Histopathological alterations included an increase in the incidence of multifocal necrosis, inflammation, pigmentation, biliary hyperplasia and the foci of cellular alteration of the liver and chronic nephropathy and protein cast of the kidney. At 20 ppm, decreases in HCT and MCV and increases in the liver and kidney weights were observed in both genders. A decrease in the HB of females and an increase in the PLT count of females were also observed. Histopathological alterations included slight increases in the incidences of hepatic necrosis, hepatic inflammation and chronic nephropathy. At 5 ppm, we found decreases in the MCV of males and the HB of females, as well as an increase in the liver weight of both genders. In the present experimental conditions, the target organs were determined to be the liver, kidney and blood cells in rats. The no-observed-adverse-effect level was considered to be <5 ppm/6 h/day and the low-observed-adverse-effect level was believed to be 5 ppm/6 h/day in rats. PMID- 17921240 TI - Investigations on permeation of mitomycin C through double layers of natural rubber gloves. AB - Treating peritoneal carcinomatosis by the aggressive cytoreductive surgery with the hyperthermic intraoperative intraperitoneal chemotherapy (HIPEC) surgeons expose their gloved hands for up to 90 min to a peritoneal dialysis solution (PDS) containing mitomycin C (MMC). We investigated the permeation of MMC through the material of three different natural rubber gloves under conditions similar to the in-use during HIPEC as well as under worst-case exposure scenario. Two different methods, a two-chamber diffusion cell and a single-chamber glass chamber method, were used to demonstrate the permeation capability. The permeation of MMC dissolved in 0.9% NaCl solution and PDS through double natural rubber glove material was tested over 2 h using four concentrations (c = 0.004, 0.008, 0.016 and 0.4 mg ml(-1)) and three receptor fluids (0.9% NaCl solution, PDS and a novel artificial sweat). In none of four glass chamber experiments and in only one of 40 diffusion cell experiments was permeation through glove material detected. The permeation occurred between 15 and 30 min under worst-case exposure scenario at a approximately 100-fold higher MMC concentration than under in-use conditions during HIPEC. The double-layer natural rubber gloves tested were effective to prevent a permeation of MMC in vitro under HIPEC-similar exposure. Our results support the glove wearing procedure in our university hospital. However, occupational exposure to antineoplastic drugs should be minimized, since there is insufficient knowledge regarding harmful effects from a long-term exposure to low doses. PMID- 17921243 TI - Family functioning and adolescents' emotional and behavioral problems: when a parent has cancer. AB - BACKGROUND: This article focuses on possible relationships between functioning of adolescents with a parent diagnosed with cancer 1-5 years earlier and family environment. PATIENTS AND METHODS: In all, 138 patients, 114 spouses and 221 adolescents completed the Family Environment Scale. Additionally, adolescents filled in the Impact of Event Scale and Youth Self-report and parents reported on the adolescents' functioning using the Child Behavior Checklist. RESULTS: Patients and spouses reported that their families differ from the norm; they are more expressive and social, better organized, less controlling and have less conflict. Adolescents reported the same and additionally find their family. Family environment was weakly to moderately strongly negatively related to the adolescents' functioning; family relationships related more strongly to the adolescents' functioning than family structure did. No significant relationship was found between family environment and the adolescents' cancer-related distress. Discrepancy in reports of family environment between parents and between parents and adolescents, in general, did not relate to the adolescents' functioning or distress. Parent-adolescent discrepancy only correlated with adolescent self-reports of their functioning. CONCLUSION: Families with parental cancer functioned positively. Despite this, family functioning seems to be a risk factor for behavioral and emotional problems in adolescents. PMID- 17921242 TI - Emerging therapeutic options for breast cancer chemotherapy during pregnancy. AB - BACKGROUND: Breast cancer is the commonest solid tumor observed during pregnancy. Anthracycline-based chemotherapy is feasible during the 2nd and 3rd trimesters of pregnancy, but few data are available on recent and highly active drugs taxanes, vinorelbine and anti-HER-2 agents in this setting. PATIENTS AND METHODS: We carried out a comprehensive review of reports documenting the use of taxanes, vinorelbine, trastuzumab and lapatinib during pregnancy in the English literature, in order to evaluate their safety profile in pregnant patients. RESULTS: Twenty-four pregnancies are described, in which no grade 3-4 maternal toxicity nor malformation in the offspring was reported. Whereas only one report studied the pharmacokinetics of paclitaxel (Taxol) during pregnancy, several preclinical reports indicate that the placental P-glycoprotein could prevent the transplacental transfer of taxanes and vinorelbine. The use of trastuzumab was associated with the occurrence of anhydramnios in three of six cases. CONCLUSION: The administration of recent drugs taxanes and vinorelbine seems feasible during the 2nd and 3rd trimesters of pregnancy, with a favorable toxicity profile. In contrast, anti-HER-2 agents may obscure the normal development of the fetal kidney, and should be avoided during pregnancy. PMID- 17921244 TI - ESMO clinical recommendations: a practical guide for medical oncologists. PMID- 17921245 TI - Application of mobile phone technology for managing chemotherapy-associated side effects. AB - BACKGROUND: Novel mobile phone technology linked to a server that communicates patients' symptoms to healthcare professionals has been adapted to register the side- effects of chemotherapy and provide advice on management of toxicity. We report a feasibility study to examine the utility of home monitoring of patients' symptoms via a mobile phone. METHODS: Six colon cancer patients receiving adjuvant chemotherapy, entered symptom data onto user friendly screens on a mobile phone twice daily. This 'real time' self assessment of nausea, vomiting, mucositis, diarrhoea and hand-foot syndrome and measurement of temperature was sent via a secured connection to a remote computer. In the event of moderate or severe symptoms (generating amber and red alerts respectively), the nurse was immediately alerted by the computer, via a pager. The nurse then contacted the patient to reinforce the automatic advice sent to the patient on their phone and to assess the patient using clinical algorithms. RESULTS: The patient used the mobile phones during the first two cycles of chemotherapy. The data were successfully analysed by the server software and alerts were generated alerting the study nurses to patients' symptoms at the appropriate time. There were 91 alerts-54 red and 37 amber; 54% (29/54) of the red alerts were data delay and transmission problems which were swiftly rectified. The remaining red alerts were managed appropriately by the study nurses. Both patients and staff felt confident in this approach to symptom management. CONCLUSIONS: This study demonstrates that the technology for monitoring patients' symptoms worked well. The patients felt secure in the knowledge that their symptoms were being closely monitored and that they were participating effectively in their own care management. PMID- 17921246 TI - Declining p53 function in the aging process: a possible mechanism for the increased tumor incidence in older populations. AB - Cancer is a disease of aging. The accumulation of mutations in individual cells over a lifetime is thought to be the reason. In this work, we explored an additional hypothesis: could p53 function decline with age, which would contribute to an enhanced mutation frequency and tumorigenesis in the aging process? The efficiency of the p53 response to gamma-irradiation was found to decline significantly in various tissues of aging mice from several inbred strains, including lower p53 transcriptional activity and p53-dependent apoptosis. This decline resulted from a decreased stabilization of the p53 protein after stress. The function of the Ataxia-telangiectasia mutated (ATM) kinase declined significantly with age, which may then be responsible for the decline of the p53 response to radiation. Declining p53 responses to other stresses were also observed in the cultured splenocytes from aging mice. Interestingly, the time of onset of this decreased p53 response correlated with the life span of mice; mice that live longer delay their onset of decreased p53 activity with time. These results suggest an enhanced fixation of mutations in older individuals because of the declining fidelity of p53-mediated apoptosis or senescence in response to stress, and they suggest a plausible explanation for the correlation between tumorigenesis and the aging process. PMID- 17921247 TI - A glycosyltransferase with a length-controlling activity as a mechanism to regulate the size of polysaccharides. AB - Cyclic beta-1,2-glucans (CbetaG) are osmolyte homopolysaccharides with a cyclic beta-1,2-backbone of 17-25 glucose residues present in the periplasmic space of several bacteria. Initiation, elongation, and cyclization, the three distinctive reactions required for building the cyclic structure, are catalyzed by the same protein, the CbetaG synthase. The initiation activity catalyzes the transference of the first glucose from UDP-glucose to a yet-unidentified amino acid residue in the same protein. Elongation proceeds by the successive addition of glucose residues from UDP-glucose to the nonreducing end of the protein-linked beta-1,2 oligosaccharide intermediate. Finally, the protein-linked intermediate is cyclized, and the cyclic glucan is released from the protein. These reactions do not explain, however, the mechanism by which the number of glucose residues in the cyclic structure is controlled. We now report that control of the degree of polymerization (DP) is carried out by a beta-1,2-glucan phosphorylase present at the CbetaG synthase C-terminal domain. This last activity catalyzes the phosphorolysis of the beta-1,2-glucosidic bond at the nonreducing end of the linear protein-linked intermediate, releasing glucose 1-phosphate. The DP is thus regulated by this "length-controlling" phosphorylase activity. To our knowledge, this is the first description of a control of the DP of homopolysaccharides. PMID- 17921248 TI - A mouse model of conditional lipodystrophy. AB - Lipodystrophies are syndromes of adipose tissue degeneration associated with severe defects in lipid and glucose homeostasis. We report here the generation and analysis of Pparg(ldi), a targeted allele that confers conditional dominant lipodystrophy in mice. The Pparg(ldi) allele was generated by insertion of the Tet activator (tTA) and a tTA-regulated Flag-Pparg1 transgene into the Pparg gene. Unexpectedly, tTA elicits mild lipodystrophy, insulin resistance, and dyslipidemia, and the Flag-PPARgamma1 transgene surprisingly exacerbates these traits. Doxycycline can both completely prevent and reverse these phenotypes, providing a mouse model of inducible lipodystrophy. Embryonic fibroblasts from either Pparg(ldi/+) or the phenotypically similar aP2-nSrebp1c (Sr) transgenic mice undergo robust adipogenesis, suggesting that neither strain develops lipodystrophy because of defective adipocyte differentiation. In addition, Pparg(ldi/+) adipose tissue shares extensive gene expression aberrations with that of Sr mice, authenticating the phenotype at the molecular level and revealing a common expression signature of lipodystrophic fat. Thus, the Pparg(ldi/+) mouse provides a conditional animal model for studying lipodystrophy and its associated physiology and gene expression. PMID- 17921249 TI - CRF-CRF1 system activation mediates withdrawal-induced increases in nicotine self administration in nicotine-dependent rats. AB - Nicotine, the main psychoactive ingredient of tobacco, induces negative emotional symptoms during abstinence that contribute to a profound craving for nicotine. However, the neurobiological mechanisms underlying how nicotine produces dependence remains poorly understood. We demonstrate one mechanism for both the anxiety-like symptoms of withdrawal and excessive nicotine intake observed after abstinence, through recruitment of the extrahypothalamic stress peptide corticotropin-releasing factor (CRF) system and activation of CRF(1) receptors. Overactivation of the CRF-CRF(1) system may contribute to nicotine dependence and may represent a prominent target for investigating the vulnerability to tobacco addiction. PMID- 17921250 TI - Kinetic evidence for a ligand-binding-induced conformational transition in the T cell receptor. AB - Thermodynamics and kinetics of the interaction between T cell receptor specific for cytomegalovirus peptide (TCR(CMV)) and its specific ligand, pp65-HLA-A*0201 complex, were studied by surface plasmon resonance and stopped-flow methods. In the latter measurements, fluorescence resonance energy transfer (FRET) between fluorescently labeled reactants was used. Thermodynamic data derived from surface plasmon resonance measurements suggest that the complex formation is driven by both favorable enthalpy and entropy. Two reaction phases were resolved by the stopped-flow measurements. The rate constant of the first step was calculated to be close to the diffusion-controlled limit rate (3x10(5) to 10(6) M(-1) s(-1)), whereas the second step's reaction rate was found to be concentration independent and relatively slow (2-4 s(-1) at 25 degrees C). These findings strongly suggest that the interactions between the TCR and its ligand, the peptide-MHC complex, proceed by a two-step mechanism, in which the second step is an induced-fit process, rate determining for antigen recognition by TCR. PMID- 17921251 TI - Impaired glutathione synthesis in schizophrenia: convergent genetic and functional evidence. AB - Schizophrenia is a complex multifactorial brain disorder with a genetic component. Convergent evidence has implicated oxidative stress and glutathione (GSH) deficits in the pathogenesis of this disease. The aim of the present study was to test whether schizophrenia is associated with a deficit of GSH synthesis. Cultured skin fibroblasts from schizophrenia patients and control subjects were challenged with oxidative stress, and parameters of the rate-limiting enzyme for the GSH synthesis, the glutamate cysteine ligase (GCL), were measured. Stressed cells of patients had a 26% (P = 0.002) decreased GCL activity as compared with controls. This reduction correlated with a 29% (P < 0.001) decreased protein expression of the catalytic GCL subunit (GCLC). Genetic analysis of a trinucleotide repeat (TNR) polymorphism in the GCLC gene showed a significant association with schizophrenia in two independent case-control studies. The most common TNR genotype 7/7 was more frequent in controls [odds ratio (OR) = 0.6, P = 0.003], whereas the rarest TNR genotype 8/8 was three times more frequent in patients (OR = 3.0, P = 0.007). Moreover, subjects with disease-associated genotypes had lower GCLC protein expression (P = 0.017), GCL activity (P = 0.037), and GSH contents (P = 0.004) than subjects with genotypes that were more frequent in controls. Taken together, the study provides genetic and functional evidence that an impaired capacity to synthesize GSH under conditions of oxidative stress is a vulnerability factor for schizophrenia. PMID- 17921252 TI - A multifunctional desaturase involved in the biosynthesis of the processionary moth sex pheromone. AB - The sex pheromone of the female processionary moth, Thaumetopoea pityocampa, is a unique C16 enyne acetate that is biosynthesized from palmitic acid. Three consecutive desaturation reactions transform this saturated precursor into the triunsaturated fatty acyl intermediate: formation of (Z)-11-hexadecenoic acid, acetylenation to 11-hexadecynoic acid, and final Delta(13) desaturation to (Z)-13 hexadecen-11-ynoic acid. By using degenerate primers common to all reported insect desaturases, a single cDNA sequence was isolated from total RNA of T. pityocampa female pheromone glands. The full-length transcript of this putative desaturase was expressed in elo1Delta/ole1Delta yeast mutants (both elongase 1 and Delta(9) desaturase-deficient) for functional assays. The construct fully rescued the Deltaole1 yeast phenotype, confirming its desaturase activity. Analysis of the unsaturated products from transformed yeast extracts demonstrated that the cloned enzyme showed Delta(11) desaturase, Delta(11) acetylenase, and Delta(13) desaturase activities. Therefore, this single desaturase may account for the three desaturation steps involved in the sex pheromone biosynthetic pathway of the processionary moth. PMID- 17921253 TI - Complement factor H deficiency in aged mice causes retinal abnormalities and visual dysfunction. AB - Age-related macular degeneration is the most common form of legal blindness in westernized societies, and polymorphisms in the gene encoding complement factor H (CFH) are associated with susceptibility to age-related macular degeneration in more than half of affected individuals. To investigate the relationship between complement factor H (CFH) and retinal disease, we performed functional and anatomical analysis in 2-year-old CFH-deficient (cfh(-/-)) mice. cfh(-/-) animals exhibited significantly reduced visual acuity and rod response amplitudes on electroretinography compared with age-matched controls. Retinal imaging by confocal scanning laser ophthalmoscopy revealed an increase in autofluorescent subretinal deposits in the cfh(-/-) mice, whereas the fundus and vasculature appeared normal. Examination of tissue sections showed an accumulation of complement C3 in the neural retina of the cfh(-/-) mice, together with a decrease in electron-dense material, thinning of Bruch's membrane, changes in the cellular distribution of retinal pigment epithelial cell organelles, and disorganization of rod photoreceptor outer segments. Collectively, these data show that, in the absence of any specific exogenous challenge to the innate immune system, CFH is critically required for the long-term functional health of the retina. PMID- 17921254 TI - Quantitative genetic correlation between trait and preference supports a sexually selected sperm process. AB - Sperm show patterns of rapid and divergent evolution that are characteristic of sexual selection. Sperm competition has been proposed as an important selective agent in the evolution of sperm morphology. However, several comparative analyses have revealed evolutionary associations between sperm length and female reproductive tract morphology that suggest patterns of male-female coevolution. In the dung beetle Onthophagus taurus, males with short sperm have a fertilization advantage that depends on the size of the female's sperm storage organ, the spermatheca; large spermathecae select for short sperm. Sperm length is heritable and is genetically correlated with male condition. Here we report significant additive genetic variation and heritability for spermatheca size and genetic covariance between spermatheca size and sperm length predicted by both the "good-sperm" and "sexy-sperm" models of postcopulatory female preference. Our data thus provide quantitative genetic support for the role of a sexually selected sperm process in the evolutionary divergence of sperm morphology, in much the same manner as precopulatory female preferences drive the evolutionary divergence of male secondary sexual traits. PMID- 17921255 TI - Structural basis for antiactivation in bacterial quorum sensing. AB - Bacteria can communicate via diffusible signal molecules they generate and release to coordinate their behavior in response to the environment. Signal molecule concentration is often proportional to bacterial population density, and when this reaches a critical concentration, reflecting a bacterial quorum, specific behaviors including virulence, symbiosis, and horizontal gene transfer are activated. Quorum-sensing regulation in many Gram-negative bacteria involves acylated homoserine lactone signals that are perceived through binding to LuxR type, acylated-homoserine-lactone-responsive transcription factors. Bacteria of the rhizobial group employ the LuxR-type transcriptional activator TraR in quorum sensing, and its activity is further regulated through interactions with the TraM antiactivator. In this study, we have crystallographically determined the 3D structure of the TraR-TraM antiactivation complex from Rhizobium sp. strain NGR234. Unexpectedly, the antiactivator TraM binds to TraR at a site distinct from its DNA-binding motif and induces an allosteric conformational change in the protein, thereby preventing DNA binding. Structural analysis reveals a highly conserved TraR-TraM interface and suggests a mechanism for antiactivation complex formation. This structure may inform alternative strategies to control quorum sensing-regulated microbial activity including amelioration of infectious disease and antibiotic resistance. In addition, the structural basis of antiactivation presents a regulatory interaction that provides general insights relevant to the field of transcription regulation and signal transduction. PMID- 17921258 TI - Effects of 7 hemoglobin variants on the measurement of glycohemoglobin by 14 analytical methods. AB - BACKGROUND: Hemoglobin variants (Hb(VAR)) are not uncommon in the Korean population, with Hb G-Coushatta and Hb Queens being the 2 most common Hb(VAR). Hb G-Coushatta is also the most common Hb(VAR) in Chinese people from the Silk Road region, as well as in some North American Indian tribes. However, data are scarce on the effect of these Hb(VAR) on the different methods used for analyzing HbA(1c). METHODS: Specimens from 24 individuals with 7 Hb(VAR) (Hb G-Coushatta, Hb Queens, Hb G-Hsi-Tsou, Hb Ube-4, Hb G-Waimanalo, Hb Inglewood, and Hb Bologna St.Orsola) were collected and tested using the International Federation of Clinical Chemistry primary reference method as well as 14 routine HbA(1c) assay methods. RESULTS: Hb G-Coushatta showed a clinically significant effect on the measured HbA(1c), particularly when analysis was performed with ion-exchange HPLC methods with short elution times. This interference could be resolved by measuring the HbA(1c) using other methods such as HPLC with a long elution time, immunoassay, boronate affinity chromatography, and enzymatic assay. Hb Queens showed a clinically significant difference, defined as a >10% deviation from regression lines, in results from the 2 HPLC methods but not in the other methods. The remaining 5 rare Hb(VAR) showed different HbA(1c) results in the different assays. CONCLUSION: Hb G-Coushatta, Hb Queens, and other rare Hb(VAR) can interfere with glycohemoglobin assays, including ion-exchange HPLC methods with short elution times, but the interference can be resolved using other unaffected methods. It is important to identify these Hb(VAR) through a careful inspection of the chromatograms and apply other noninterfering methods for accurate measurements of the HbA(1c). PMID- 17921256 TI - Variant estrogen receptor-c-Src molecular interdependence and c-Src structural requirements for endothelial NO synthase activation. AB - Little is known about the tyrosine kinase c-Src's function in the systemic circulation, in particular its role in arterial responses to hormonal stimuli. In human aortic and venous endothelial cells, c-Src is indispensable for 17beta estradiol (E2)-stimulated phosphatidylinositol 3-kinase/Akt/endothelial NO synthase (eNOS) pathway activation, a possible mechanism in E2-mediated vascular protection. Here we show that c-Src supports basal and E2-stimulated NO production and is required for E2-induced vasorelaxation in murine aortas. Only membrane c-Src is structurally and functionally involved in E2-induced eNOS activation. Independent of c-Src kinase activity, c-Src is associated with an N terminally truncated estrogen receptor alpha variant (ER46) and eNOS in the plasma membrane through its "open" (substrate-accessible) conformation. In the presence of E2, c-Src kinase is activated by membrane ER46 and in turn phosphorylates ER46 for subsequent ER46 and c-Src membrane recruitment, the assembly of an eNOS-centered membrane macrocomplex, and membrane-initiated eNOS activation. Overall, these results provide insights into a critical role for the tyrosine kinase c-Src in estrogen-stimulated arterial responses, and in membrane initiated rapid signal transduction, for which obligate complex assembly and localization require the c-Src substrate-accessible structure. PMID- 17921259 TI - Modified form of the fibrinogen Bbeta chain (des-Gln Bbeta), a potential long lived marker of pancreatitis. AB - BACKGROUND: During an investigation of genetic variants of fibrinogen, we observed a novel form of the Bbeta chain, with a mass decrease of approximately 128 Da, in one of the controls. The plasma sample originated from an individual who had experienced acute pancreatitis a week earlier but whose serum amylase activity had returned to normal. We investigated the structure of the modified fibrinogen and explored its relationship to pancreatic disease. METHOD: Fibrinogen was isolated from the plasma of 9 individuals with increased pancreatic amylase activity (114-1826 U/L) and presumed pancreatitis and from 6 control individuals with amylase activities <56 U/L. Fibrinogen (or fibrin) Bbeta chains were isolated by reversed-phase HPLC and analyzed directly by electrospray ionization mass spectrometry. Tryptic and CNBr peptide mapping and thrombin treatment pinpointed the location of the 128-Da loss in mass. RESULTS: The acquired fibrinogen Bbeta chain modification was attributable to the loss of its C-terminal glutamine residue. Incubating purified fibrinogen with pancreatic carboxypeptidase A (CpA) produced an identical modification. The des-Gln Bbeta fibrinogen accounted for >80% of the Bbeta chains in 3 of the individuals with increased amylase but only approximately 5% of the Bbeta chains in control samples. CONCLUSION: Pancreatic CpA activity is used as an index of acute pancreatic disease, but given that the circulatory half-lives of fibrinogen and CpA are approximately 4 days and only 2.5 h, respectively, measuring des-Gln Bbeta fibrinogen, the in vivo product of CpA activity, could provide clinicians with retrospective evidence of disease. PMID- 17921257 TI - Comparing active and repressed expression states of genes controlled by the Polycomb/Trithorax group proteins. AB - Drosophila Polycomb group (PcG) and Trithorax group (TrxG) proteins are responsible for the maintenance of stable transcription patterns of many developmental regulators, such as the homeotic genes. We have used ChIP-on-chip to compare the distribution of several PcG/TrxG proteins, as well as histone modifications in active and repressed genes across the two homeotic complexes ANT C and BX-C. Our data indicate the colocalization of the Polycomb repressive complex 1 (PRC1) with Trx and the DNA binding protein Pleiohomeotic (Pho) at discrete sequence elements as well as significant chromatin assembly differences in active and inactive regions. Trx binds to the promoters of active genes and noncoding transcripts. Most strikingly, in the active state, Pho covers extended chromatin domains over many kilobases. This feature of Pho, observed on many polytene chromosome puffs, reflects a previously undescribed function. At the hsp70 gene, we demonstrate in mutants that Pho is required for transcriptional recovery after heat shock. Besides its presumptive function in recruiting PcG complexes to their site of action, our results now uncover that Pho plays an additional role in the repression of already induced genes. PMID- 17921260 TI - Buprenorphine and norbuprenorphine in hair of pregnant women and their infants after controlled buprenorphine administration. AB - BACKGROUND: Buprenorphine is under investigation as a pharmacotherapeutic agent for treating opioid dependence in pregnant women. We hypothesized that there would be a relationship between the cumulative maternal dose of buprenorphine during pregnancy and the concentration of buprenorphine and norbuprenorphine in maternal and infant hair. METHODS: This study examined buprenorphine and norbuprenorphine concentrations in hair obtained from 9 buprenorphine-maintained pregnant women and 4 of their infants. Specimens were analyzed by liquid chromatography-tandem mass spectrometry with limits of quantification of 3.0 pg/mg. All maternal hair specimens were washed with methylene chloride before analysis, and when sufficient amounts of maternal hair were available, specimens also were analyzed without washing. Infant hair specimens were not washed. RESULTS: Buprenorphine concentrations were significantly greater in unwashed hair than washed hair (P = 0.031). Norbuprenorphine concentrations were significantly greater than buprenorphine concentrations in both maternal (P = 0.0097) and infant hair (P = 0.0033). There were statistically significant associations between the cumulative maternal dose of buprenorphine administered and the concentrations of buprenorphine (washed, P <0.0001; unwashed, P = 0.0004), norbuprenorphine (washed, P <0.0001; unwashed, P = 0.0005), and buprenorphine plus norbuprenorphine (washed, P <0.0001; unwashed, P = 0.0005) for both washed and unwashed maternal hair specimens. There was a significant positive association between concentrations of buprenorphine and norbuprenorphine in maternal hair (washed, P <0.0001; unwashed, P = 0.0003), a trend for this association in infant hair (P = 0.08), and an association between buprenorphine concentrations in maternal unwashed hair and infant hair (P = 0.0002). The buprenorphine:norbuprenorphine ratio increased in distal segments. CONCLUSION: Buprenorphine treatment during gestation provides an opportunity for monitoring drug disposition in maternal and fetal tissues under controlled conditions. PMID- 17921261 TI - Detailed technical analysis of urine RNA-based tumor diagnostics reveals ETS2/urokinase plasminogen activator to be a novel marker for bladder cancer. AB - BACKGROUND: The noninvasive detection of RNA tumor markers in body fluids represents an attractive diagnostic option, but diagnostic performance of tissue derived markers is often poorer when measured in body fluids rather than in tumors. We aimed to develop a procedure for measurement of tumor RNA in urine that would minimize donor-dependent influences on the results. METHODS: RNA isolated from urinary cell pellet, cell-depleted fraction, and whole urine was quantified by reverse transcription quantitative-PCR. The donor-dependent influence of urine background on individual steps of the standardized procedure was analyzed using an external RNA standard. Using a test set of samples from 61 patients with bladder cancer and 37 healthy donors, we compared 4 putative RNA tumor markers identified in whole urine with 5 established, tissue-derived RNA tumor markers for the detection of bladder cancer. RESULTS: Of the markers analyzed by this system, the RNA ratio of v-ets erythroblastosis virus E26 oncogene homolog 2 (avian; ETS2) to urokinase plasminogen activator (uPA) enabled the most specific (100%) and sensitive (75.4%) detection of bladder cancer from whole urine, with an area under the curve of 0.929 (95% CI 0.882-0.976). CONCLUSIONS: The described methodology for RNA marker detection in urine appears to be clinically applicable. The ratio of ETS2 mRNA to uPA mRNA in urine is a potential marker for bladder cancer. PMID- 17921262 TI - The Listeria monocytogenes homolog of the Escherichia coli era gene is involved in adhesion to inert surfaces. AB - Two transposon-insertional mutants of Listeria monocytogenes showing smaller viable surface-attached cell populations after disinfection with N,N-didecyl-N,N dimethylammonium chloride were identified. In both mutants, transposon Tn917-lac was found to be inserted into the same gene, lmo1462, which is homologous to the essential Escherichia coli era gene. Both L. monocytogenes lmo1462-disrupted mutants displayed lower growth rates, as was also shown for several E. coli era mutants, and the lmo1462 gene was able to complement the growth defect of an E. coli era mutant. We showed that the disruption of lmo1462 decreased the ability of L. monocytogenes cells to adhere to stainless steel. Our results suggest that this era-like gene is involved in adhesion and contributes to the presence of L. monocytogenes on surfaces. PMID- 17921263 TI - Improvement of NADPH-dependent bioconversion by transcriptome-based molecular breeding. AB - Transcriptome data for a xylitol-producing recombinant Escherichia coli were obtained and used to tune up its productivity. Structural genes of NADPH dependent D-xylose reductase and D-xylose permease were inserted into an Escherichia coli chromosome to construct a recombinant strain producing xylitol from D-xylose for use as a model system for NADPH-dependent bioconversion. Transcriptome analysis of xylitol-producing and nonproducing conditions for the recombinant revealed that xylitol production down-regulated 56 genes. These genes were then selected as candidate factors for suppression of the NADPH supply and were disrupted to validate their functions. Of the gene disruptants, that resulting from the deletion of yhbC showed the best bioconversion rate. Also, the deletion accelerated cell growth during log phase. The features of the mutant could be maintained in jar fermenter-scale production of xylitol. Thus, our novel molecular host strain breeding method using transcriptome analysis was fully effective and could be applied to improving various industrial strains. PMID- 17921264 TI - DNA fingerprinting of lactic acid bacteria in sauerkraut fermentations. AB - Previous studies using traditional biochemical identification methods to study the ecology of commercial sauerkraut fermentations revealed that four species of lactic acid bacteria, Leuconostoc mesenteroides, Lactobacillus plantarum, Pediococcus pentosaceus, and Lactobacillus brevis, were the primary microorganisms in these fermentations. In this study, 686 isolates were collected from four commercial fermentations and analyzed by DNA fingerprinting. The results indicate that the species of lactic acid bacteria present in sauerkraut fermentations are more diverse than previously reported and include Leuconostoc citreum, Leuconostoc argentinum, Lactobacillus paraplantarum, Lactobacillus coryniformis, and Weissella sp. The newly identified species Leuconostoc fallax was also found. Unexpectedly, only two isolates of P. pentosaceus and 15 isolates of L. brevis were recovered during this study. A better understanding of the microbiota may aid in the development of low-salt fermentations, which may have altered microflora and altered sensory characteristics. PMID- 17921265 TI - Detection of bacteriophage-infected cells of Lactococcus lactis by using flow cytometry. AB - Bacteriophage infection in dairy fermentation constitutes a serious problem worldwide. We have studied bacteriophage infection in Lactococcus lactis by using the flow cytometer. The first effect of the infection of the bacterium is a change from cells in chains toward single cells. We interpret this change as a consequence of a cease in cell growth, while the ongoing cell divisions leave the cells as single cells. Late in the infection cycle, cells with low-density cell walls appear, and these cells can be detected on cytograms of light scatter versus, for instance, fluorescence of stained DNA. We describe a new method for detection of phage infection in Lactococcus lactis dairy cultures. The method is based on flow cytometric detection of cells with low-density cell walls. The method allows fast and early detection of phage-infected bacteria, independently of which phage has infected the culture. The method can be performed in real time and therefore increases the chance of successful intervention in the fermentation process. PMID- 17921266 TI - Growth characteristics of Brevibacterium, Corynebacterium, Microbacterium, and Staphylococcus spp. isolated from surface-ripened cheese. AB - The growth characteristics of five bacteria, Brevibacterium aurantiacum 1-16-58, Corynebacterium casei DPC 5298(T), Corynebacterium variabile DPC 5310, Microbacterium gubbeenense DPC 5286(T), and Staphylococcus saprophyticus 4E61, all of which were isolated from the surface of smear cheese, were studied in complex and chemically defined media. All of the coryneforms, except M. gubbeenense, grew in 12% salt, while B. aurantiacum and S. saprophyticus grew in 15% salt. All five bacteria assimilated lactate in a semisynthetic medium, and none of the coryneform bacteria assimilated lactose. Glucose assimilation was poor, except by S. saprophyticus and C. casei. Five to seven amino acids were assimilated by the coryneforms and 12 by S. saprophyticus. Glutamate, phenylalanine, and proline were utilized by all five bacteria, whereas utilization of serine, threonine, aspartate, histidine, alanine, arginine, leucine, isoleucine, and glycine depended on the organism. Growth of C. casei restarted after addition of glutamate, proline, serine, and lactate at the end of the exponential phase, indicating that these amino acids and lactate can be used as energy sources. Pantothenic acid was essential for the growth of C. casei and M. gubbeenense. Omission of biotin reduced the growth of B. aurantiacum, C. casei, and M. gubbeenense. All of the bacteria contained lactate dehydrogenase activity (with both pyruvate and lactate as substrates) and glutamate pyruvate transaminase activity but not urease activity. PMID- 17921267 TI - Novel Cryptosporidium genotype in wild Australian mice (Mus domesticus). AB - A total of 250 mouse fecal specimens collected from crop farms in Queensland, Australia, were screened for the presence of Cryptosporidium spp. using PCR. Of these, 19 positives were detected and characterized at a number of loci, including the 18S rRNA gene, the acetyl coenzyme A gene, and the actin gene. Sequence and phylogenetic analyses identified two genotypes: mouse genotype I and a novel genotype (mouse genotype II), which is likely to be a valid species. Cryptosporidium parvum, which is zoonotic, was not detected. The results of the study indicate that wild Australian mice that are not in close contact with livestock are probably not an important reservoir of Cryptosporidium infection for humans and other animals. PMID- 17921268 TI - Genomic and biochemical studies demonstrating the absence of an alkane-producing phenotype in Vibrio furnissii M1. AB - Vibrio furnissii M1 was recently reported to biosynthesize n-alkanes when grown on biopolymers, sugars, or organic acids (M. O. Park, J. Bacteriol. 187:1426 1429, 2005). In the present study, V. furnissii M1 was subjected to genomic analysis and studied biochemically. The sequence of the 16S rRNA gene and repetitive PCR showed that V. furnissii M1 was not identical to other V. furnissii strains tested, but the level of relatedness was consistent with its assignment as a V. furnissii strain. Pulsed-field gel electrophoresis showed chromosomal bands at approximately 3.2 and 1.8 Mb, similar to other Vibrio strains. Complete genomic DNA from V. furnissii M1 was sequenced with 21-fold coverage. Alkane biosynthetic and degradation genes could not be identified. Moreover, V. furnissii M1 did not produce demonstrable levels of n-alkanes in vivo or in vitro. In vivo experiments were conducted by growing V. furnissii M1 under different conditions, extracting with solvent, and analyzing extracts by gas chromatography-mass spectrometry. A highly sensitive assay was used for in vitro experiments with cell extracts and [(14)C]hexadecanol. The data are consistent with the present strain being a V. furnissii with properties similar to those previously described but lacking the alkane-producing phenotype. V. furnissii ATCC 35016, also reported to biosynthesize alkanes, was found in the present study not to produce alkanes. PMID- 17921269 TI - Genome-wide screen of Salmonella genes expressed during infection in pigs, using in vivo expression technology. AB - Pigs are a food-producing species that readily carry Salmonella but, in the great majority of cases, do not show clinical signs of disease. Little is known about the functions required by Salmonella to be maintained in pigs. We have devised a recombinase-based promoter-trapping strategy to identify genes with elevated expression during pig infection with Salmonella enterica serovar Typhimurium. A total of 55 clones with in vivo-induced promoters were selected from a genomic library of approximately 10,000 random Salmonella DNA fragments fused to the recombinase cre, and the cloned DNA fragments were analyzed by sequencing. Thirty one genes encoding proteins involved in bacterial adhesion and colonization (including bcfA, hscA, rffG, and yciR), virulence (metL), heat shock (hscA), and a sensor of a two-component regulator (hydH) were identified. Among the 55 clones, 19 were isolated from both the tonsils and the intestine, while 23 were identified only in the intestine and 13 only in tonsils. High temperature and increased osmolarity were identified as environmental signals that induced in vivo-expressed genes, suggesting possible signals for expression. PMID- 17921270 TI - Variability of total and pathogenic Vibrio parahaemolyticus densities in northern Gulf of Mexico water and oysters. AB - Vibrio parahaemolyticus is indigenous to coastal environments and a frequent cause of seafood-borne gastroenteritis in the United States, primarily due to raw oyster consumption. Previous seasonal-cycle studies of V. parahaemolyticus have identified water temperature as the strongest environmental predictor. Salinity has also been identified, although it is evident that its effect on annual variation is not as pronounced. The effects of other environmental factors, both with respect to the seasonal cycle and intraseasonal variation, are uncertain. This study investigated intraseasonal variations of densities of total and pathogenic V. parahaemolyticus organisms in oysters and overlying waters during the summer of 2004 at two sites in the northern Gulf of Mexico. Regression analyses indicated significant associations (P < 0.001) between total V. parahaemolyticus densities and salinity, as well as turbidity in water and in oysters at the Mississippi site but not at the Alabama site. Pathogenic V. parahaemolyticus organisms in Mississippi oyster and water samples were detected in 56% (9 out of 16) and 78% (43 out of 55) of samples, respectively. In contrast, 44% (7 out of 16) of oyster samples and 30% (14 out of 47) of water samples from Alabama were positive. At both sites, there was greater sample-to sample variability in pathogenic V. parahaemolyticus densities than in total V. parahaemolyticus densities. These data suggest that, although total V. parahaemolyticus densities may be very informative, there is greater uncertainty when total V. parahaemolyticus densities are used to predict the risk of infection by pathogenic V. parahaemolyticus than previously recognized. PMID- 17921271 TI - Distribution and replication of the pathogenicity plasmid pPATH in diverse populations of the gall-forming bacterium Pantoea agglomerans. AB - Pantoea agglomerans has been transformed from a commensal bacterium into two related gall-forming pathovars by acquisition of pPATH plasmids containing a pathogenicity island (PAI). This PAI harbors an hrp/hrc gene cluster, type III effectors, and phytohormone biosynthetic genes. DNA typing by pulsed-field gel electrophoresis revealed two major groups of P. agglomerans pv. gypsophilae and one group of P. agglomerans pv. betae. The pPATH plasmids of the different groups had nearly identical replicons (98% identity), and the RepA protein showed the highest level of similarity with IncN plasmid proteins. A series of plasmids, designated pRAs, in which the whole replicon region (2,170 bp) or deleted derivatives of it were ligated with nptI were generated for replicon analysis. A basic 929-bp replicon (pRA6) was sufficient for replication in Escherichia coli and in nonpathogenic P. agglomerans. However, the whole replicon region (pRA1) was necessary for expulsion of the pPATH plasmid, which resulted in the loss of pathogenicity. The presence of direct repeats in the replicon region suggests that the pPATH plasmid is an iteron plasmid and that the repeats may regulate its replication. The pPATH plasmids are nonconjugative but exhibit a broad host range, as shown by replication of pRA1 in Erwinia, Pseudomonas, and Xanthomonas. Restriction fragment length polymorphism analyses indicated that the PAIs in the two groups of P. agglomerans pv. gypsophilae are similar but different from those in P. agglomerans pv. betae. The results could indicate that the pPATH plasmids evolved from a common ancestral mobilizable plasmid that was transferred into different strains of P. agglomerans. PMID- 17921272 TI - Molecular analysis of the diversity of sulfate-reducing and sulfur-oxidizing prokaryotes in the environment, using aprA as functional marker gene. AB - The dissimilatory adenosine-5'-phosphosulfate reductase is a key enzyme of the microbial sulfate reduction and sulfur oxidation processes. Because the alpha- and beta-subunit-encoding genes, aprBA, are highly conserved among sulfate reducing and sulfur-oxidizing prokaryotes, they are most suitable for molecular profiling of the microbial community structure of the sulfur cycle in environment. In this study, a new aprA gene-targeting assay using a combination of PCR and denaturing gradient gel electrophoresis is presented. The screening of sulfate-reducing and sulfur-oxidizing reference strains as well as the analyses of environmental DNA from diverse habitats (e.g., microbial mats, invertebrate tissue, marine and estuarine sediments, and filtered hydrothermal water) by the new primer pair revealed an improved microbial diversity coverage and less pronounced template-to-PCR product bias in direct comparison to those of the previously published primer set (B. Deplancke, K. R. Hristova, H. A. Oakley, V. J. McCracken, R. Aminov, R. I. Mackie, and H. R. Gaskins, Appl. Environ. Microbiol. 66:2166-2174, 2000). The concomitant molecular detection of sulfate reducing and sulfur-oxidizing prokaryotes was confirmed. The new assay was applied in comparison with the 16S rRNA gene-based analysis to investigate the microbial diversity of the sulfur cycle in sediment, seawater, and manganese crust samples from four study sites in the area of the Lesser Antilles volcanic arc, Caribbean Sea (Caribflux project). The aprA gene-based approach revealed putative sulfur-oxidizing Alphaproteobacteria of chemolithoheterotrophic lifestyle to have been abundant in the nonhydrothermal sediment and water column. In contrast, the sulfur-based microbial community that inhabited the surface of the volcanic manganese crust was more complex, consisting predominantly of putative chemolithoautotrophic sulfur oxidizers of the Betaproteobacteria and Gammaproteobacteria. PMID- 17921273 TI - Culture-dependent and -independent characterization of microbial communities associated with a shallow submarine hydrothermal system occurring within a coral reef off Taketomi Island, Japan. AB - Microbial communities in a shallow submarine hydrothermal system near Taketomi Island, Japan, were investigated using cultivation-based and molecular techniques. The main hydrothermal activity occurred in a craterlike basin (depth, approximately 23 m) on the coral reef seafloor. The vent fluid (maximum temperature, >52 degrees C) contained 175 microM H2S and gas bubbles mainly composed of CH4 (69%) and N2 (29%). A liquid serial dilution cultivation technique targeting a variety of metabolism types quantified each population in the vent fluid and in a white microbial mat located near the vent. The most abundant microorganisms cultivated from both the fluid and the mat were autotrophic sulfur oxidizers, including mesophilic Thiomicrospira spp. and thermophilic Sulfurivirga caldicuralii. Methane oxidizers were the second most abundant organisms in the fluid; one novel type I methanotroph exhibited optimum growth at 37 degrees C, and another novel type I methanotroph exhibited optimum growth at 45 degrees C. The number of hydrogen oxidizers cultivated only from the mat was less than the number of sulfur and methane oxidizers, although a novel mesophilic hydrogen-oxidizing member of the Epsilonproteobacteria was isolated. Various mesophilic to hyperthermophilic heterotrophs, including sulfate-reducing Desulfovibrio spp., iron-reducing Deferribacter sp., and sulfur-reducing Thermococcus spp., were also cultivated. Culture-independent 16S rRNA gene clone analysis of the vent fluid and mat revealed highly diverse archaeal communities. In the bacterial community, S. caldicuralii was identified as the predominant phylotype in the fluid (clonal frequency, 25%). Both bacterial clone libraries indicated that there were bacterial communities involved in sulfur, hydrogen, and methane oxidation and sulfate reduction. Our results indicate that there are unique microbial communities that are sustained by active chemosynthetic primary production rather than by photosynthetic production in a shallow hydrothermal system where sunlight is abundant. PMID- 17921275 TI - Chromosomal and symbiotic relationships of rhizobia nodulating Medicago truncatula and M. laciniata. AB - Multilocus sequence typing (MLST) is a sequence-based method used to characterize bacterial genomes. This method was used to examine the genetic structure of Medicago-nodulating rhizobia at the Amra site, which is located in an arid region of Tunisia. Here the annual medics Medicago laciniata and M. truncatula are part of the natural flora. The goal of this study was to identify whether distinct chromosomal groups of rhizobia nodulate M. laciniata because of its restricted requirement for specific rhizobia. The MLST analysis involved determination of sequence variation in 10 chromosomal loci of 74 isolates each of M. laciniata and M. truncatula. M. truncatula was used as a control trap host, because unlike M. laciniata, it has relatively unrestricted rhizobial requirements. Allelic diversity among the plasmid nodC alleles in the isolates was also determined. The 148 isolates were placed into 26 chromosomal sequence types (STs), only 3 of which had been identified previously. The rhizobia of M. laciniata were shown to be part of the general Medicago-nodulating population in the soil because 99.95% of the isolates had chromosomal genotypes similar to those recovered from M. truncatula. However, the isolates recovered from M. laciniata were less diverse than those recovered from M. truncatula, and they also harbored an unusual nodC allele. This could perhaps be best explained by horizontal transfer of the different nodC alleles among members of the Medicago-nodulating rhizobial population at the field site. Evidence indicating a history of lateral transfer of rhizobial symbiotic genes across distinct chromosomal backgrounds is provided. PMID- 17921276 TI - Analysis of nitrogen-fixing members of the epsilon subclass of Proteobacteria in salt marsh sediments. AB - Based on phylogenetic analysis of clones retrieved from two nifH gene clone libraries that were created using cDNA from suboxic sediment samples obtained from areas densely vegetated with the high-salt marsh plant Spartina patens, a primer set was designed to target nitrogen-fixing bacteria with sequence similarities to members of the epsilon subclass of Proteobacteria. Nested PCR, denaturing gel electrophoresis, and subsequent sequence analysis of reamplified fragments confirmed the specificity of the primer set by retrieving nifH sequences of only putative members of the epsilon subclass of Proteobacteria, all of which were characterized by a highly divergent 27- or 36-bp insertion in both DNA and cDNA. PMID- 17921277 TI - Sequence characterization and comparative analysis of three plasmids isolated from environmental Vibrio spp. AB - The horizontal transfer of genes by mobile genetic elements such as plasmids and phages can accelerate genome diversification of Vibrio spp., affecting their physiology, pathogenicity, and ecological character. In this study, sequence analysis of three plasmids from Vibrio spp. previously isolated from salt marsh sediment revealed the remarkable diversity of these elements. Plasmids p0908 (81.4 kb), p23023 (52.5 kb), and p09022 (31.0 kb) had a predicted 99, 64, and 32 protein-coding sequences and G+C contents of 49.2%, 44.7%, and 42.4%, respectively. A phylogenetic tree based on concatenation of the host 16S rRNA and rpoA nucleotide sequences indicated p23023 and p09022 were isolated from strains most closely related to V. mediterranei and V. campbellii, respectively, while the host of p0908 forms a clade with V. fluvialis and V. furnissii. Many predicted proteins had amino acid identities to proteins of previously characterized phages and plasmids (24 to 94%). Predicted proteins with similarity to chromosomally encoded proteins included RecA, a nucleoid-associated protein (NdpA), a type IV helicase (UvrD), and multiple hypothetical proteins. Plasmid p0908 had striking similarity to enterobacteria phage P1, sharing genetic organization and amino acid identity for 23 predicted proteins. This study provides evidence of genetic exchange between Vibrio plasmids, phages, and chromosomes among diverse Vibrio spp. PMID- 17921278 TI - Expanding the Helicobacter pylori genetic toolbox: modification of an endogenous plasmid for use as a transcriptional reporter and complementation vector. AB - Helicobacter pylori is an important human pathogen. However, the study of this organism is often limited by a relative shortage of genetic tools. In an effort to expand the methods available for genetic study, an endogenous H. pylori plasmid was modified for use as a transcriptional reporter and as a complementation vector. This was accomplished by addition of an Escherichia coli origin of replication, a kanamycin resistance cassette, a promoterless gfpmut3 gene, and a functional multiple cloning site to form pTM117. The promoters of amiE and pfr, two well-characterized Fur-regulated promoters, were fused to the promoterless gfpmut3, and green fluorescent protein (GFP) expression of the fusions in wild-type and delta fur strains was analyzed by flow cytometry under iron-replete and iron-depleted conditions. GFP expression was altered as expected based on current knowledge of Fur regulation of these promoters. RNase protection assays were used to determine the ability of this plasmid to serve as a complementation vector by analyzing amiE, pfr, and fur expression in wild-type and delta fur strains carrying a wild-type copy of fur on the plasmid. Proper regulation of these genes was restored in the delta fur background under high- and low-iron conditions, signifying complementation of both iron-bound and apo Fur regulation. These studies show the potential of pTM117 as a molecular tool for genetic analysis of H. pylori. PMID- 17921279 TI - Effect of CO2 on colony development by Bifidobacterium species. AB - This report investigates the requirement for CO2 for colony formation by Bifidobacterium species in both anoxic and oxic environments. All tested Bifidobacterium species exhibited difficulty in developing colonies in an atmosphere of 100% N2 but developed well when 1% CO2 was present. In the presence of CO2, the oxygen tolerance of the tested species was not improved. In the absence of CO2, only B. boum, a microaerophilic species, could develop colonies under an N2-based 5% O2 atmosphere, indicating that while CO2 is not an essential factor for colony development, both CO2 and O2 have stimulatory effects on B. boum colony development. PMID- 17921280 TI - Transient marker system for iterative gene targeting of a prototrophic fungus. AB - Auxotrophic microorganisms are often used for genetic engineering, because their biosynthetic deficiency can be complemented by the transforming DNA and allows selection for transformants that have become prototrophic. However, when complementation is obtained by ectopic expression this may lead to unpredictable side effects on the phenotype and, consequently, misinterpretation of experimental data. There are various ways to overcome the problem of auxotrophy, but the most reliable is to restore the function of the defective biosynthetic gene at the native genomic locus. This can be done by either sexual crossing or further genetic engineering. For fungal species lacking a perfect state or situations in which gene targeting is generally cumbersome we have developed a concept that allows transient disruption of pyrG. When the gene is in the disrupted state, multiple rounds of gene targeting can be performed with the strain. Once the desired genome engineering is completed, pyrG function can be rapidly returned to wild type by a simple selection scheme. PMID- 17921274 TI - Metagenomic characterization of Chesapeake Bay virioplankton. AB - Viruses are ubiquitous and abundant throughout the biosphere. In marine systems, virus-mediated processes can have significant impacts on microbial diversity and on global biogeocehmical cycling. However, viral genetic diversity remains poorly characterized. To address this shortcoming, a metagenomic library was constructed from Chesapeake Bay virioplankton. The resulting sequences constitute the largest collection of long-read double-stranded DNA (dsDNA) viral metagenome data reported to date. BLAST homology comparisons showed that Chesapeake Bay virioplankton contained a high proportion of unknown (homologous only to environmental sequences) and novel (no significant homolog) sequences. This analysis suggests that dsDNA viruses are likely one of the largest reservoirs of unknown genetic diversity in the biosphere. The taxonomic origin of BLAST homologs to viral library sequences agreed well with reported abundances of cooccurring bacterial subphyla within the estuary and indicated that cyanophages were abundant. However, the low proportion of Siphophage homologs contradicts a previous assertion that this family comprises most bacteriophage diversity. Identification and analyses of cyanobacterial homologs of the psbA gene illustrated the value of metagenomic studies of virioplankton. The phylogeny of inferred PsbA protein sequences suggested that Chesapeake Bay cyanophage strains are endemic in that environment. The ratio of psbA homologous sequences to total cyanophage sequences in the metagenome indicated that the psbA gene may be nearly universal in Chesapeake Bay cyanophage genomes. Furthermore, the low frequency of psbD homologs in the library supports the prediction that Chesapeake Bay cyanophage populations are dominated by Podoviridae. PMID- 17921281 TI - Epidemiology, relative invasive ability, molecular characterization, and competitive performance of Campylobacter jejuni strains in the chicken gut. AB - One hundred forty-one Campylobacter jejuni isolates from humans with diarrhea and 100 isolates from retailed poultry meat were differentiated by flaA typing. The bacteria were isolated in a specific geographical area (Dunedin) in New Zealand over a common time period. Twenty nine flaA types were detected, one of which (flaA restriction fragment length polymorphism type 15 [flaA-15]) predominated among isolates from humans ( approximately 30% of isolates). This strain was of low prevalence (5% of isolates) among poultry isolates. flaA-15 strains were five to six times more invasive of HEp2 cells in an in vitro assay than a flaA type (flaA-3) that was commonly encountered on poultry meat (23% of isolates) but was seldom associated with human illness (5%). Competitive-exclusion experiments with chickens, utilizing real-time quantitative PCR to measure the population sizes of specific strains representing flaA-15 (T1016) and flaA-3 (Pstau) in digesta, were carried out. These experiments showed that T1016 always outcompeted Pstau in the chicken intestine. Genomic comparisons of T1016 and Pstau were made using DNA microarrays representing the genome of C. jejuni NCTC 11168. These comparisons revealed differences between the strains in the gene content of the Cj1417c-to Cj1442c region of the genome, which is associated with the formation of capsular polysaccharide. The strains differed in Penner type (T1016, O42; Pstau, O53). It was concluded that poultry meat was at least one source of human infection with C. jejuni, that some Campylobacter strains detected in poultry meat are of higher virulence for humans than others, and that bacterial attributes affecting strain virulence and commensal colonization ability may be linked. PMID- 17921282 TI - The cyclic AMP receptor protein modulates colonial morphology in Vibrio cholerae. AB - Inactivation of the quorum-sensing regulator HapR causes Vibrio cholerae El Tor biotype strain C7258 to adopt a rugose colonial morphology that correlates with enhanced biofilm formation. V. cholerae mutants lacking the cyclic AMP (cAMP) receptor protein (CRP) produce very little HapR, which results in elevated expression of Vibrio exopolysaccharide (vps) genes and biofilm compared to the wild type. However, Deltacrp mutants still exhibited smooth colonial morphology and expressed reduced levels of vps genes compared to isogenic hapR mutants. In this study we demonstrate that deletion of crp and cya (adenylate cyclase) converts a rugose DeltahapR mutant to a smooth one. The smooth DeltahapR Deltacrp and DeltahapR Deltacya double mutants could be converted back to rugose by complementation with crp and cya, respectively. CRP was found to enhance the expression of VpsR, a strong activator of vps expression, but to diminish transcription of VpsT. Ectopic expression of VpsR in smooth DeltahapR Deltacrp and DeltahapR Deltacya double mutants restored rugose colonial morphology. Lowering intracellular cAMP levels in a DeltahapR mutant by the addition of glucose diminished VpsR expression and colonial rugosity. On the basis of our results, we propose a model for the regulatory input of CRP on exopolysaccharide biosynthesis. PMID- 17921283 TI - N-(3-hydroxyhexanoyl)-l-homoserine lactone is the biologically relevant quormone that regulates the phz operon of Pseudomonas chlororaphis strain 30-84. AB - Phenazine production by Pseudomonas fluorescens 2-79 and P. chlororaphis isolates 30-84 and PCL1391 is regulated by quorum sensing through the activator PhzR and acyl-homoserine lactones (acyl-HSLs) synthesized by PhzI. PhzI from P. fluorescens 2-79 produces five acyl-HSLs that include four 3-hydroxy species. Of these, N-(3-hydroxyhexanoyl)-HSL is the biologically relevant ligand for PhzR. The quorum-sensing systems of P. chlororaphis strains 30-84 and PCL1391 have been reported to produce and respond to N-(hexanoyl)-HSL. These differences were of interest since PhzI and PhzR of strain 2-79 share almost 90% sequence identity with orthologs from strains 30-84 and PCL1391. In this study, as assessed by thin layer chromatography, the three strains produce almost identical complements of acyl-HSLs. The major species produced by P. chlororaphis 30-84 were identified by mass spectrometry as 3-OH-acyl-HSLs with chain lengths of 6, 8, and 10 carbons. Heterologous bacteria expressing cloned phzI from strain 30-84 produced the four 3-OH acyl-HSLs in amounts similar to those seen for the wild type. Strain 30-84, but not strain 2-79, also produced N-(butanoyl)-HSL. A second acyl-HSL synthase of strain 30-84, CsaI, is responsible for the synthesis of this short-chain signal. Strain 30-84 accumulated N-(3-OH-hexanoyl)-HSL to the highest levels, more than 100-fold greater than that of N-(hexanoyl)-HSL. In titration assays, PhzR(30-84) responded to both N-(3-OH-hexanoyl)- and N-(hexanoyl)-HSL with equal sensitivities. However, only the 3-OH-hexanoyl signal is produced by strain 30-84 at levels high enough to activate PhzR. We conclude that strains 2-79, 30-84, and PCL1391 use N-(3-OH-hexanoyl)-HSL to activate PhzR. PMID- 17921284 TI - Comparison of cyanopeptolin genes in Planktothrix, Microcystis, and Anabaena strains: evidence for independent evolution within each genus. AB - The major cyclic peptide cyanopeptolin 1138, produced by Planktothrix strain NIVA CYA 116, was characterized and shown to be structurally very close to the earlier characterized oscillapeptin E. A cyanopeptolin gene cluster likely to encode the corresponding peptide synthetase was sequenced from the same strain. The 30-kb oci gene cluster contains two novel domains previously not detected in nonribosomal peptide synthetase gene clusters (a putative glyceric acid activating domain and a sulfotransferase domain), in addition to seven nonribosomal peptide synthetase modules. Unlike in two previously described cyanopeptolin gene clusters from Anabaena and Microcystis, a halogenase gene is not present. The three cyanopeptolin gene clusters show similar gene and domain arrangements, while the binding pocket signatures deduced from the adenylation domain sequences and the additional tailoring domains vary. This suggests loss and gain of tailoring domains within each genus, after the diversification of the three clades, as major events leading to the present diversity. The ABC transporter genes associated with the cyanopeptolin gene clusters form a monophyletic clade and accordingly are likely to have evolved as part of the functional unit. Phylogenetic analyses of adenylation and condensation domains, including domains from cyanopeptolins and microcystins, show a closer similarity between the Planktothrix and Microcystis cyanopeptolin domains than between these and the Anabaena domain. No clear evidence of recombination between cyanopeptolins and microcystins could be detected. There were no strong indications of horizontal gene transfer of cyanopeptolin gene sequences across the three genera, supporting independent evolution within each genus. PMID- 17921285 TI - Quantitative distributions of Epsilonproteobacteria and a Sulfurimonas subgroup in pelagic redoxclines of the central Baltic Sea. AB - Members of the class Epsilonproteobacteria are known to be of major importance in biogeochemical processes at oxic-anoxic interfaces. In pelagic redoxclines of the central Baltic Sea, an uncultured epsilonproteobacterium related to Sulfurimonas denitrificans was proposed to play a key role in chemolithotrophic denitrification (I. Brettar, M. Labrenz, S. Flavier, J. Botel, H. Kuosa, R. Christen, and M. G. Hofle, Appl. Environ. Microbiol. 72:1364-1372, 2006). In order to determine the abundance, activity, and vertical distribution of this bacterium in high-resolution profiles, 16S rRNA cloning and catalyzed reporter deposition and fluorescence in situ hybridization (CARD-FISH) and quantitative PCR measurements were carried out. The results showed that 21% of the derived clone sequences, which in the present study were grouped together under the name GD17, had >99% similarity to the uncultured epsilonproteobacterium. A specific gene probe against GD17 (S-*-Sul-0090-a-A-18) was developed and used for enumeration by CARD-FISH. In different pelagic redoxclines sampled during August 2003, May 2005, and February 2006, GD17 cells were always detected from the lower oxic area to the sulfidic area. Maximal abundance was detected around the chemocline, where sulfide and nitrate concentrations were close to the detection limit. The highest GD17 numbers (2 x 10(5) cells ml(-1)), representing up to 15% of the total bacteria, were comparable to those reported for Epsilonproteobacteria in pelagic redoxclines of the Black Sea and the Cariaco Trench (X. Lin, S. G. Wakeham, I. F. Putnam, Y. M. Astor, M. I. Scranton, A. Y. Chistoserdov, and G. T. Taylor, Appl. Environ. Microbiol. 72:2679-2690, 2006). However, in the Baltic Sea redoxclines, Epsilonproteobacteria consisted nearly entirely of cells belonging to the distinct GD17 group. This suggested that GD17 was the best-adapted epsilonproteobacterium within this ecological niche. PMID- 17921286 TI - Biosurfactant production and surface translocation are regulated by PlcR in Bacillus cereus ATCC 14579 under low-nutrient conditions. AB - Bacillus cereus ATCC 14579 can respond to nutrient changes by adopting different forms of surface translocation. The B. cereus ATCC 14579 DeltaplcR mutant, but not the wild type, formed dendritic (branched) patterns on EPS [a low-nutrient medium that contains 7.0 g K(2)HPO(4), 3.0 g KH(2)PO(4), 0.1 g MgSO(4).7H(2)O, 0.1 g (NH(4))(2)SO(4), 0.01 g CaCl(2), 0.001 g FeSO(4), 0.1 g NaCl, 1.0 g glucose, and 125 mg yeast extract per liter] containing 0.7% agar. The dendritic patterns formed by sliding translocation of nonflagellated cells are enhanced under low-nutrient conditions and require sufficient production of a biosurfactant, which appears to be repressed by PlcR. The wild-type and complemented strains failed to slide on the surface of EPS agar because of the production of low levels of biosurfactant. Precoating EPS agar surfaces with surfactin (a biosurfactant produced by Bacillus subtilis) or biosurfactant purified from the DeltaplcR mutant rescued the ability of the wild-type and complemented strains to slide. When grown on a nutrient-rich medium like Luria Bertani agar, both the wild-type and DeltaplcR mutant strains produced flagella. The wild type was hyperflagellated and elongated and exhibited swarming behavior, while the DeltaplcR mutant was multiflagellated and the cells often formed long chains but did not swarm. Thin-layer chromatography and mass spectrometry analyses suggested that the biosurfactant purified from the DeltaplcR mutant was a lipopeptide and had a mass of 1,278.1722 (m/z). This biosurfactant has hemolytic activity and inhibited the growth of several gram-positive bacteria. PMID- 17921287 TI - Identification and characterization of the dps promoter of Mycobacterium smegmatis: promoter recognition by stress-specific extracytoplasmic function sigma factors sigmaH and sigmaF. AB - The survival of a bacterium with a depleted oxygen or nutrient supply is important for its long-term persistence inside the host under stressful conditions. We studied a gene, dps, from Mycobacterium smegmatis, encoding a protein, Dps (for DNA binding protein from starved cells), which is overexpressed under oxidative and nutritional stresses and provides bimodal protection to the bacterial DNA. Characterization of the dps promoter in vivo is therefore important. We cloned a 1-kb putative promoter region of the dps gene of M. smegmatis in an Escherichia coli-Mycobacterium shuttle vector, pSD5B, immediately upstream of the lacZ gene. Promoter activities were assayed in vivo both in solid medium and in liquid cultures by quantitative beta-galactosidase activity measurements. To characterize the minimal promoter region, a 200-bp fragment from the whole 1-kb sequence was further cloned in the same vector, and in a similar way, beta-galactosidase activity was quantitated. Primer extension analysis was performed to determine the +1 transcription start site of the gene. Point mutations were inserted in the putative promoter sequences in the -10 and -20 regions, and the promoter sequence was confirmed. The promoter was not recognized by purified M. smegmatis core RNA polymerase reconstituted with purified Mycobacterium tuberculosis sigmaA or sigmaB during multiple- and single-round in vitro transcription assays. Promoter-specific in vivo pull-down assays with an immobilized 1-kb DNA fragment containing the dps promoter established that extracellular function sigma factors were associated with this starvation inducible promoter. Single-round transcription at the dps promoter further supported the idea that only core RNA polymerase reconstituted with sigmaF or sigmaH can generate proper transcripts. PMID- 17921288 TI - Response of Desulfovibrio vulgaris to alkaline stress. AB - The response of exponentially growing Desulfovibrio vulgaris Hildenborough to pH 10 stress was studied using oligonucleotide microarrays and a study set of mutants with genes suggested by microarray data to be involved in the alkaline stress response deleted. The data showed that the response of D. vulgaris to increased pH is generally similar to that of Escherichia coli but is apparently controlled by unique regulatory circuits since the alternative sigma factors (sigma S and sigma E) contributing to this stress response in E. coli appear to be absent in D. vulgaris. Genes previously reported to be up-regulated in E. coli were up-regulated in D. vulgaris; these genes included three ATPase genes and a tryptophan synthase gene. Transcription of chaperone and protease genes (encoding ATP-dependent Clp and La proteases and DnaK) was also elevated in D. vulgaris. As in E. coli, genes involved in flagellum synthesis were down-regulated. The transcriptional data also identified regulators, distinct from sigma S and sigma E, that are likely part of a D. vulgaris Hildenborough-specific stress response system. Characterization of a study set of mutants with genes implicated in alkaline stress response deleted confirmed that there was protective involvement of the sodium/proton antiporter NhaC-2, tryptophanase A, and two putative regulators/histidine kinases (DVU0331 and DVU2580). PMID- 17921289 TI - Evolution and functional characterization of the RH50 gene from the ammonia oxidizing bacterium Nitrosomonas europaea. AB - The family of ammonia and ammonium channel proteins comprises the Amt proteins, which are present in all three domains of life with the notable exception of vertebrates, and the homologous Rh proteins (Rh50 and Rh30) that have been described thus far only in eukaryotes. The existence of an RH50 gene in bacteria was first revealed by the genome sequencing of the ammonia-oxidizing bacterium Nitrosomonas europaea. Here we have used a phylogenetic approach to study the evolution of the N. europaea RH50 gene, and we show that this gene, probably as a component of an integron cassette, has been transferred to the N. europaea genome by horizontal gene transfer. In addition, by functionally characterizing the Rh50(Ne) protein and the corresponding knockout mutant, we determined that NeRh50 can mediate ammonium uptake. The RH50(Ne) gene may thus have replaced functionally the AMT gene, which is missing in the genome of N. europaea and may be regarded as a case of nonorthologous gene displacement. PMID- 17921290 TI - Characterization of the Mycobacterium tuberculosis 4-diphosphocytidyl-2-C-methyl D-erythritol synthase: potential for drug development. AB - Mycobacterium tuberculosis utilizes the methylerythritol phosphate (MEP) pathway for biosynthesis of isopentenyl diphosphate and its isomer, dimethylallyl diphosphate, precursors of all isoprenoid compounds. This pathway is of interest as a source of new drug targets, as it is absent from humans and disruption of the responsible genes has shown a lethal phenotype for Escherichia coli. In the MEP pathway, 4-diphosphocytidyl-2-C-methyl-D-erythritol is formed from 2-C-methyl D-erythritol 4-phosphate (MEP) and CTP in a reaction catalyzed by a 4 diphosphocytidyl-2-C-methyl-D-erythritol synthase (IspD). In the present work, we demonstrate that Rv3582c is essential for M. tuberculosis: Rv3582c has been cloned and expressed, and the encoded protein has been purified. The purified M. tuberculosis IspD protein was capable of catalyzing the formation of 4 diphosphocytidyl-2-C-methyl-D-erythritol in the presence of MEP and CTP. The enzyme was active over a broad pH range (pH 6.0 to 9.0), with peak activity at pH 8.0. The activity was absolutely dependent upon divalent cations, with 20 mM Mg2+ being optimal, and replacement of CTP with other nucleotide 5'-triphosphates did not support activity. Under the conditions tested, M. tuberculosis IspD had Km values of 58.5 microM for MEP and 53.2 microM for CTP. Calculated kcat and kcat/Km values were 0.72 min(-1) and 12.3 mM(-1) min(-1) for MEP and 1.0 min(-1) and 18.8 mM(-1) min(-1) for CTP, respectively. PMID- 17921291 TI - Orotate phosphoribosyltransferase from Corynebacterium ammoniagenes lacking a conserved lysine. AB - The pyrE gene, encoding orotate phosphoribosyltransferase (OPRTase), was cloned by nested PCR and colony blotting from Corynebacterium ammoniagenes ATCC 6872, which is widely used in nucleotide production. Sequence analysis shows that there is a lack of an important conserved lysine (Lys 73 in Salmonella enterica serovar Typhimurium OPRTase) in the C. ammoniagenes OPRTase. This lysine has been considered to contribute to the initiation of catalysis. The enzyme was overexpressed and purified from a recombinant Escherichia coli strain. The molecular mass of the purified OPRTase was determined to be 45.4 +/- 1.5 kDa by gel filtration. Since the molecular mass for the subunit of the enzyme was 21.3 +/- 0.6 kDa, the native enzyme exists as a dimer. Divalent magnesium was necessary for the activity of the enzyme and can be substituted for by Mn2+ and Co2+. The optimal pH for the forward (phosphoribosyl transfer) reaction is 10.5 to 11.5, which is higher than that of other reported OPRTases, and the optimal pH for the reverse (pyrophosphorolysis) reaction is 5.5 to 6.5. The Km values for the four substrates were determined to be 33 microM for orotate, 64 microM for 5 phosphoribosyl-1-pyrophosphate (PRPP), 45 microM for orotidine-5-phosphate (OMP), and 36 microM for pyrophosphate. The Km value for OMP is much larger than those of other organisms. These differences may be due to the absence of Lys 73, which is present in the active sites of other OPRTases and is known to interact with OMP and PRPP. PMID- 17921292 TI - Functional analysis of the M.HpyAIV DNA methyltransferase of Helicobacter pylori. AB - A large number of genes encoding restriction-modification (R-M) systems are found in the genome of the human pathogen Helicobacter pylori. R-M genes comprise approximately 10% of the strain-specific genes, but the relevance of having such an abundance of these genes is not clear. The type II methyltransferase (MTase) M.HpyAIV, which recognizes GANTC sites, was present in 60% of the H. pylori strains analyzed, whereof 69% were resistant to restriction enzyme digestion, which indicated the presence of an active MTase. H. pylori strains with an inactive M.HpyAIV phenotype contained deletions in regions of homopolymers within the gene, which resulted in premature translational stops, suggesting that M.HpyAIV may be subjected to phase variation by a slipped-strand mechanism. An M.HpyAIV gene mutant was constructed by insertional mutagenesis, and this mutant showed the same viability and ability to induce interleukin-8 in epithelial cells as the wild type in vitro but had, as expected, lost the ability to protect its self-DNA from digestion by a cognate restriction enzyme. The M.HpyAIV from H. pylori strain 26695 was overexpressed in Escherichia coli, and the protein was purified and was able to bind to DNA and protect GANTC sites from digestion in vitro. A bioinformatic analysis of the number of GANTC sites located in predicted regulatory regions of H. pylori strains 26695 and J99 resulted in a number of candidate genes. katA, a selected candidate gene, was further analyzed by quantitative real-time reverse transcription-PCR and shown to be significantly down-regulated in the M.HpyAIV gene mutant compared to the wild-type strain. This demonstrates the influence of M.HpyAIV methylation in gene expression. PMID- 17921293 TI - Control of the transcription of a short gene encoding a cyclic peptide in Streptococcus thermophilus: a new quorum-sensing system? AB - Gram-positive bacteria secrete a variety of peptides that are often subjected to posttranslational modifications and that are either antimicrobials or pheromones involved in bacterial communication. Our objective was to identify peptides secreted by Streptococcus thermophilus, a nonpathogenic bacterium widely used in dairy technology in association with other bacteria, and to understand their potential roles in cell-cell communication. Using reverse-phase liquid chromatography, mass spectrometry, and Edman sequencing, we analyzed the culture supernatants of three S. thermophilus strains (CNRZ1066, LMG18311, and LMD-9) grown in a medium containing no peptides. We identified several peptides in the culture supernatants, some of them found with the three strains while others were specific to the LMD-9 strain. We focused our study on a new modified peptide secreted by S. thermophilus LMD-9 and designated Pep1357C. This peptide contains 9 amino acids and lost 2 Da in a posttranslational modification, most probably a dehydrogenation, leading to a linkage between the Lys2 and Trp6 residues. Production of Pep1357C and transcription of its encoding gene depend on both the medium composition and the growth phase. Furthermore, we demonstrated that transcription of the gene coding for Pep1357C is drastically decreased in mutants inactivated for the synthesis of a short hydrophobic peptide, a transcriptional regulator, or the oligopeptide transport system. Taken together, our results led us to deduce that the transcription of the Pep1357C-encoding gene is controlled by a new quorum-sensing system. PMID- 17921294 TI - Multiple superoxide dismutases in Agrobacterium tumefaciens: functional analysis, gene regulation, and influence on tumorigenesis. AB - Agrobacterium tumefaciens possesses three iron-containing superoxide dismutases (FeSods) encoded by distinct genes with differential expression patterns. SodBI and SodBII are cytoplasmic isozymes, while SodBIII is a periplasmic isozyme. sodBI is expressed at a high levels throughout all growth phases. sodBII expression is highly induced upon exposure to superoxide anions in a SoxR dependent manner. sodBIII is expressed only during stationary phase. Analysis of the physiological function of sods reveals that the inactivation of sodBI markedly reduced levels of resistance to a superoxide generator, menadione. A mutant lacking all three Sod enzymes is the most sensitive to menadione treatment, indicating that all sods contribute at various levels towards the overall menadione resistance level. Sods also have important roles in A. tumefaciens virulence toward a host plant. A sodBI but not a sodBII or sodBIII mutant showed marked reduction in its ability to induce tumors on tobacco leaf discs, while the triple sod null mutant is avirulent. PMID- 17921295 TI - Full activation of Enterococcus faecalis gelatinase by a C-terminal proteolytic cleavage. AB - Enterococci account for nearly 10% of all nosocomial infections and constitute a significant treatment challenge due to their multidrug resistance properties. One of the well-studied virulence factors of Enterococcus faecalis is a secreted bacterial protease, termed gelatinase, which has been shown to contribute to the process of biofilm formation. Gelatinase belongs to the M4 family of bacterial zinc metalloendopeptidases, typified by thermolysin. Gelatinase is synthesized as a preproenzyme consisting of a signal sequence, a putative propeptide, and then the mature enzyme. We determined that the molecular mass of the mature protein isolated from culture supernatant was 33,030 Da, which differed from the predicted molecular mass, 34,570 Da, by over 1,500 Da. Using N-terminal sequencing, we confirmed that the mature protein begins at the previously identified sequence VGSEV, thus suggesting that the 1,500-Da molecular mass difference resulted from a C-terminal processing event. By using mutants with site-directed mutations within a predicted C-terminal processing site and mutants with C-terminal deletions fused to a hexahistidine tag, we determined that the processing site is likely to be between residues D304 and I305 and that it requires the Q306 residue. The results suggest that the E. faecalis gelatinase requires C-terminal processing for full activation of protease activity, making it a unique enzyme among the members of the M4 family of proteases of gram positive bacteria. PMID- 17921296 TI - Mapping critical interactive sites within the periplasmic domain of the Vibrio cholerae type II secretion protein EpsM. AB - The type II secretion (T2S) system is present in many gram-negative species, both pathogenic and nonpathogenic, where it supports the delivery of a variety of toxins, proteases, and lipases into the extracellular environment. In Vibrio cholerae, the T2S apparatus is composed of 12 Eps proteins that assemble into a multiprotein complex that spans the entire cell envelope. Two of these proteins, EpsM and EpsL, are key components of the secretion machinery present in the inner membrane. In addition to likely forming homodimers, EpsL and EpsM have been shown to form a stable complex in the inner membrane and to protect each other from proteolytic degradation. To identify and map the specific regions of EpsM involved in protein-protein interactions with both another molecule of EpsM and EpsL, we tested the interactions of deletion constructs of EpsM with full-length EpsM and EpsL by functional characterization and copurification as well as coimmunoprecipitation. Analysis of the truncated EpsM mutants revealed that the region of EpsM from amino acids 100 to 135 is necessary for EpsM to form homo oligomers, while residues 84 to 99 appear to be critical for a stable interaction with EpsL. PMID- 17921297 TI - Transposon Tn7 directs transposition into the genome of filamentous bacteriophage M13 using the element-encoded TnsE protein. AB - The bacterial transposon Tn7 has a pathway of transposition that preferentially targets conjugal plasmids. We propose that this same transposition pathway recognizes a structure or complex found during filamentous bacteriophage replication, likely by targeting negative-strand synthesis. The ability to insert into both plasmid and bacteriophage DNAs that are capable of cell-to-cell transfer would help explain the wide distribution of Tn7 relatives. PMID- 17921298 TI - Influence of the poly-3-hydroxybutyrate (PHB) granule-associated proteins (PhaP1 and PhaP2) on PHB accumulation and symbiotic nitrogen fixation in Sinorhizobium meliloti Rm1021. AB - Sinorhizobium meliloti cells store excess carbon as intracellular poly-3 hydroxybutyrate (PHB) granules that assist survival under fluctuating nutritional conditions. PHB granule-associated proteins (phasins) are proposed to regulate PHB synthesis and granule formation. Although the enzymology and genetics of PHB metabolism in S. meliloti have been well characterized, phasins have not yet been described for this organism. Comparison of the protein profiles of the wild type and a PHB synthesis mutant revealed two major proteins absent from the mutant. These were identified by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) as being encoded by the SMc00777 (phaP1) and SMc02111 (phaP2) genes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins associated with PHB granules followed by MALDI-TOF confirmed that PhaP1 and PhaP2 were the two major phasins. Double mutants were defective in PHB production, while single mutants still produced PHB, and unlike PHB synthesis mutants that have reduced exopolysaccharide, the double mutants had higher exopolysaccharide levels. Medicago truncatula plants inoculated with the double mutant exhibited reduced shoot dry weight (SDW), although there was no corresponding reduction in nitrogen fixation activity. Whether the phasins are involved in a metabolic regulatory response or whether the reduced SDW is due to a reduction in assimilation of fixed nitrogen rather than a reduction in nitrogen fixation activity remains to be established. PMID- 17921299 TI - Identification of a fourth formate dehydrogenase in Methylobacterium extorquens AM1 and confirmation of the essential role of formate oxidation in methylotrophy. AB - A mutant of Methylobacterium extorquens AM1 with lesions in genes for three formate dehydrogenase (FDH) enzymes was previously described by us (L. Chistoserdova, M. Laukel, J.-C. Portais, J. A. Vorholt, and M. E. Lidstrom, J. Bacteriol. 186:22-28, 2004). This mutant had lost its ability to grow on formate but still maintained the ability to grow on methanol. In this work, we further investigated the phenotype of this mutant. Nuclear magnetic resonance experiments with [13C]formate, as well as 14C-labeling experiments, demonstrated production of labeled CO2 in the mutant, pointing to the presence of an additional enzyme or a pathway for formate oxidation. The tungsten-sensitive phenotype of the mutant suggested the involvement of a molybdenum-dependent enzyme. Whole-genome array experiments were conducted to test for genes overexpressed in the triple-FDH mutant compared to the wild type, and a gene (fdh4A) was identified whose translated product carried similarity to an uncharacterized putative molybdopterin-binding oxidoreductase-like protein sharing relatively low similarity with known formate dehydrogenase alpha subunits. Mutation of this gene in the triple-FDH mutant background resulted in a methanol-negative phenotype. When the gene was deleted in the wild-type background, the mutant revealed diminished growth on methanol with accumulation of high levels of formate in the medium, pointing to an important role of FDH4 in methanol metabolism. The identity of FDH4 as a novel FDH was also confirmed by labeling experiments that revealed strongly reduced CO2 formation in growing cultures. Mutation of a small open reading frame (fdh4B) downstream of fdh4A resulted in mutant phenotypes similar to the phenotypes of fdh4A mutants, suggesting that fdh4B is also involved in formate oxidation. PMID- 17921300 TI - A conserved glycine residue of trimeric autotransporter domains plays a key role in Yersinia adhesin A autotransport. AB - The Yersinia adhesin A (YadA) is a trimeric autotransporter adhesin of enteric yersiniae. It consists of three major domains: a head mediating adherence to host cells, a stalk involved in serum resistance, and an anchor that forms a membrane pore and is responsible for the autotransport function. The anchor contains a glycine residue, nearly invariant throughout trimeric autotransporter adhesins, that faces the pore lumen. To address the role of this glycine, we replaced it with polar amino acids of increasing side chain size and expressed wild-type and mutant YadA in Escherichia coli. The mutations did not impair the YadA-mediated adhesion to collagen and to host cells or the host cell cytokine production, but they decreased the expression levels and stability of YadA trimers with increasing side chain size. Likewise, autoagglutination and resistance to serum were decreased in these mutants. We found that the periplasmic protease DegP is involved in the degradation of YadA and that in an E. coli degP deletion strain, mutant versions of YadA were expressed almost to wild-type levels. We conclude that the conserved glycine residue affects both the export and the stability of YadA and consequently some of its putative functions in pathogenesis. PMID- 17921301 TI - The yydFGHIJ operon of Bacillus subtilis encodes a peptide that induces the LiaRS two-component system. AB - The Bacillus subtilis LiaRS two-component system (TCS) responds to perturbations of the cell envelope induced by lipid II-interacting antibiotics, such as vancomycin, ramoplanin, nisin, and bacitracin. Here, we characterize Tn7 generated mutations that induce the liaRS TCS. In addition to insertions in liaF, a known negative regulator of the LiaRS TCS, we identified two disruptions in the last two genes of the yydFGHIJ operon. This operon is predicted to encode a 49 amino-acid peptide (YydF), a modification enzyme (YydG), a membrane-embedded protease (YydH), and an ATP-binding cassette (ABC) transporter (YydIJ). Genome sequence comparisons suggest that the yydFGHIJ operon may have been acquired by horizontal transfer. Inactivation of the YydIJ transporter resulted in increased expression from the LiaR-dependent P(liaI) promoter only in the presence of the yydFGH genes. Cells harboring the complete yydFGHIJ operon induced LiaR activity in cocultured cells lacking either this transporter or the complete operon. These results suggest that this operon is involved in the synthesis and export of a modified peptide (YydF*) that elicits cell envelope stress sensed by the LiaRS TCS. PMID- 17921302 TI - Identification and characterization of the flavin:NADH reductase (PrnF) involved in a novel two-component arylamine oxygenase. AB - Two-component oxygenases catalyze a wide variety of important oxidation reactions. Recently we characterized a novel arylamine N-oxygenase (PrnD), a new member of the two-component oxygenase family (J. Lee et al., J. Biol. Chem. 280:36719-36728, 2005). Although arylamine N-oxygenases are widespread in nature, aminopyrrolnitrin N-oxygenase (PrnD) represents the only biochemically and mechanistically characterized arylamine N-oxygenase to date. Here we report the use of bioinformatic and biochemical tools to identify and characterize the reductase component (PrnF) involved in the PrnD-catalyzed unusual arylamine oxidation. The prnF gene was identified via sequence analysis of the whole genome of Pseudomonas fluorescens Pf-5 and subsequently cloned and overexpressed in Escherichia coli. The purified PrnF protein catalyzes reduction of flavin adenine dinucleotide (FAD) by NADH with a k(cat) of 65 s(-1) (K(m) = 3.2 muM for FAD and 43.1 muM for NADH) and supplies reduced FAD to the PrnD oxygenase component. Unlike other known reductases in two-component oxygenase systems, PrnF strictly requires NADH as an electron donor to reduce FAD and requires unusual protein protein interaction with the PrnD component for the efficient transfer of reduced FAD. This PrnF enzyme represents the first cloned and characterized flavin reductase component in a novel two-component arylamine oxygenase system. PMID- 17921303 TI - Spatial organization of Myxococcus xanthus during fruiting body formation. AB - Microcinematography was used to examine fruiting body development of Myxococcus xanthus. Wild-type cells progress through three distinct phases: a quiescent phase with some motility but little aggregation (0 to 8 h), a period of vigorous motility leading to raised fruiting bodies (8 to 16 h), and a period of maturation during which sporulation is initiated (16 to 48 h). Fruiting bodies are extended vertically in a series of tiers, each involving the addition of a cell monolayer on top of the uppermost layer. A pilA (MXAN_5783) mutant produced less extracellular matrix material and thus allowed closer examination of tiered aggregate formation. A csgA (MXAN_1294) mutant exhibited no quiescent phase, aberrant aggregation in phase 2, and disintegration of the fruiting bodies in the third phase. PMID- 17921304 TI - Substitution of a highly conserved histidine in the Escherichia coli heat shock transcription factor, sigma32, affects promoter utilization in vitro and leads to overexpression of the biofilm-associated flu protein in vivo. AB - The heat shock sigma factor (sigma(32) in Escherichia coli) directs the bacterial RNA polymerase to promoters of a specific sequence to form a stable complex, competent to initiate transcription of genes whose products mitigate the effects of exposure of the cell to high temperatures. The histidine at position 107 of sigma(32) is at the homologous position of a tryptophan residue at position 433 of the main sigma factor of E. coli, sigma(70). This tryptophan is essential for the strand separation step leading to the formation of the initiation-competent RNA polymerase-promoter complex. The heat shock sigma factors of all gammaproteobacteria sequenced have a histidine at this position, while in the alpha- and deltaproteobacteria, it is a tryptophan. In vitro the alanine-for histidine substitution at position 107 (H107A) destabilizes complexes between the GroE promoter and RNA polymerase containing sigma(32), implying that H107 plays a role in formation or maintenance of the strand-separated complex. In vivo, the H107A substitution in sigma(32) impedes recovery from heat shock (exposure to 42 degrees C), and it also leads to overexpression at lower temperatures (30 degrees C) of the Flu protein, which is associated with biofilm formation. PMID- 17921305 TI - Expression of the sigmaF-directed csfB locus prevents premature appearance of sigmaG activity during sporulation of Bacillus subtilis. AB - During sporulation, sigma(G) becomes active in the prespore upon the completion of engulfment. We show that the inactivation of the sigma(F)-directed csfB locus resulted in premature activation of sigma(G). CsfB exerted control distinct from but overlapping with that exerted by LonA to prevent inappropriate sigma(G) activation. The artificial induction of csfB severely compromised spore formation. PMID- 17921306 TI - Comparative genomic hybridization detects secondary chromosomal deletions in Escherichia coli K-12 MG1655 mutants and highlights instability in the flhDC region. AB - The use of whole-genome microarrays for monitoring mutagenized or otherwise engineered genetic derivatives is a potentially powerful tool for checking genomic integrity. Using comparative genomic hybridization of a number of unrelated, directed deletion mutants in Escherichia coli K-12 MG1655, we identified unintended secondary genomic deletions in the flhDC region in delta fnr, delta crp, and delta creB mutants. These deletions were confirmed by PCR and phenotypic tests. Our findings show that nonmotile progeny are found in some MG1655 directed deletion mutants, and studies on the effects of gene knockouts should be viewed with caution when the mutants have not been screened for the presence of secondary deletions or confirmed by other methods. PMID- 17921307 TI - Effects of oxygen on virulence traits of Streptococcus mutans. AB - Oxygen profoundly affects the composition of oral biofilms. Recently, we showed that exposure of Streptococcus mutans to oxygen strongly inhibits biofilm formation and alters cell surface biogenesis. To begin to dissect the underlying mechanisms by which oxygen affects known virulence traits of S. mutans, transcription profiling was used to show that roughly 5% of the genes of this organism are differentially expressed in response to aeration. Among the most profoundly upregulated genes were autolysis-related genes and those that encode bacteriocins, the ClpB protease chaperone subunit, pyruvate dehydrogenase, the tricarboxylic acid cycle enzymes, NADH oxidase enzymes, and certain carbohydrate transporters and catabolic pathways. Consistent with our observation that the ability of S. mutans to form biofilms was severely impaired by oxygen exposure, transcription of the gtfB gene, which encodes one of the primary enzymes involved in the production of water-insoluble, adhesive glucan exopolysaccharides, was down-regulated in cells growing aerobically. Further investigation revealed that transcription of gtfB, but not gtfC, was responsive to oxygen and that aeration causes major changes in the amount and degree of cell association of the Gtf enzymes. Moreover, inactivation of the VicK sensor kinase affected the expression and localization the GtfB and GtfC enzymes. This study provides novel insights into the complex transcriptional and posttranscriptional regulatory networks used by S. mutans to modulate virulence gene expression and exopolysaccharide production in response to changes in oxygen availability. PMID- 17921309 TI - TrwC-mediated site-specific recombination is controlled by host factors altering local DNA topology. AB - R388 conjugative relaxase TrwC acts as a site-specific recombinase, promoting recombination between two cognate oriTs on double-stranded DNA substrates. The relaxosome component TrwA is also required for efficient recombination. In this work we present data on the in vivo control of this reaction by host proteins that affect local DNA topology. In the absence of TrwA, binding of integration host factor (IHF) to the oriT keeps the recombination levels low, probably by keeping the relaxosome complex, formed at recombination locus 1, in a "closed" conformation. In an IHF-deficient (IHF-) background, the formation of a transcript elongation complex at this locus still hampers recombination. A mutation abating the promoter sequence at locus 1, or repression of transcription by exposure to rifampin, lifts the inhibition imposed on recombination in an IHF- background. We also observe an increase in conjugation efficiency under these conditions. Relieving the inhibition imposed by these host factors allows efficient levels of recombination between short oriT loci in the absence of TrwA. The presence of TrwA counteracts these inhibitory effects. TrwA would then activate both recombination and conjugation by switching the conformation of the relaxosome to an "open" form that exposes single-stranded DNA at the nic site, promoting the initial TrwC nicking reaction. PMID- 17921308 TI - Unusual starch degradation pathway via cyclodextrins in the hyperthermophilic sulfate-reducing archaeon Archaeoglobus fulgidus strain 7324. AB - The hyperthermophilic archaeon Archaeoglobus fulgidus strain 7324 has been shown to grow on starch and sulfate and thus represents the first sulfate reducer able to degrade polymeric sugars. The enzymes involved in starch degradation to glucose 6-phosphate were studied. In extracts of starch-grown cells the activities of the classical starch degradation enzymes, alpha-amylase and amylopullulanase, could not be detected. Instead, evidence is presented here that A. fulgidus utilizes an unusual pathway of starch degradation involving cyclodextrins as intermediates. The pathway comprises the combined action of an extracellular cyclodextrin glucanotransferase (CGTase) converting starch to cyclodextrins and the intracellular conversion of cyclodextrins to glucose 6 phosphate via cyclodextrinase (CDase), maltodextrin phosphorylase (Mal-P), and phosphoglucomutase (PGM). These enzymes, which are all induced after growth on starch, were characterized. CGTase catalyzed the conversion of starch to mainly beta-cyclodextrin. The gene encoding CGTase was cloned and sequenced and showed highest similarity to a glucanotransferase from Thermococcus litoralis. After transport of the cyclodextrins into the cell by a transport system to be defined, these molecules are linearized via a CDase, catalyzing exclusively the ring opening of the cyclodextrins to the respective maltooligodextrins. These are degraded by a Mal-P to glucose 1-phosphate. Finally, PGM catalyzes the conversion of glucose 1-phosphate to glucose 6-phosphate, which is further degraded to pyruvate via the modified Embden-Meyerhof pathway. PMID- 17921310 TI - Rhodobacter capsulatus OlsA is a bifunctional enzyme active in both ornithine lipid and phosphatidic acid biosynthesis. AB - The Rhodobacter capsulatus genome contains three genes (olsA [plsC138], plsC316, and plsC3498) that are annotated as lysophosphatidic acid (1-acyl-sn-glycerol-3 phosphate) acyltransferase (AGPAT). Of these genes, olsA was previously shown to be an O-acyltransferase in the second step of ornithine lipid biosynthesis, which is important for optimal steady-state levels of c-type cytochromes (S. Aygun Sunar, S. Mandaci, H.-G. Koch, I. V. J. Murray, H. Goldfine, and F. Daldal. Mol. Microbiol. 61:418-435, 2006). The roles of the remaining plsC316 and plsC3498 genes remained unknown. In this work, these genes were cloned, and chromosomal insertion-deletion mutations inactivating them were obtained to define their function. Characterization of these mutants indicated that, unlike the Escherichia coli plsC, neither plsC316 nor plsC3498 was essential in R. capsulatus. In contrast, no plsC316 olsA double mutant could be isolated, indicating that an intact copy of either olsA or plsC316 was required for R. capsulatus growth under the conditions tested. Compared to OlsA null mutants, PlsC316 null mutants contained ornithine lipid and had no c-type cytochrome related phenotype. However, they exhibited slight growth impairment and highly altered total fatty acid and phospholipid profiles. Heterologous expression in an E. coli plsC(Ts) mutant of either R. capsulatus plsC316 or olsA gene products supported growth at a nonpermissive temperature, exhibited AGPAT activity in vitro, and restored phosphatidic acid biosynthesis. The more vigorous AGPAT activity displayed by PlsC316 suggested that plsC316 encodes the main AGPAT required for glycerophospholipid synthesis in R. capsulatus, while olsA acts as an alternative AGPAT that is specific for ornithine lipid synthesis. This study therefore revealed for the first time that some OlsA enzymes, like the enzyme of R. capsulatus, are bifunctional and involved in both membrane ornithine lipid and glycerophospholipid biosynthesis. PMID- 17921311 TI - Structure, function, and regulation of the aldouronate utilization gene cluster from Paenibacillus sp. strain JDR-2. AB - Direct bacterial conversion of the hemicellulose fraction of hardwoods and crop residues to biobased products depends upon extracellular depolymerization of methylglucuronoxylan (MeGAX(n)), followed by assimilation and intracellular conversion of aldouronates and xylooligosaccharides to fermentable xylose. Paenibacillus sp. strain JDR-2, an aggressively xylanolytic bacterium, secretes a multimodular cell-associated GH10 endoxylanase (XynA1) that catalyzes depolymerization of MeGAX(n) and rapidly assimilates the principal products, beta 1,4-xylobiose, beta-1,4-xylotriose, and MeGAX(3), the aldotetrauronate 4-O methylglucuronosyl-alpha-1,2-xylotriose. Genomic libraries derived from this bacterium have now allowed cloning and sequencing of a unique aldouronate utilization gene cluster comprised of genes encoding signal transduction regulatory proteins, ABC transporter proteins, and the enzymes AguA (GH67 alpha glucuronidase), XynA2 (GH10 endoxylanase), and XynB (GH43 beta-xylosidase/alpha arabinofuranosidase). Expression of these genes, as well as xynA1 encoding the secreted GH10 endoxylanase, is induced by growth on MeGAX(n) and repressed by glucose. Sequences in the yesN, lplA, and xynA2 genes within the cluster and in the distal xynA1 gene show significant similarity to catabolite responsive element (cre) defined in Bacillus subtilis for recognition of the catabolite control protein (CcpA) and consequential repression of catabolic regulons. The aldouronate utilization gene cluster in Paenibacillus sp. strain JDR-2 operates as a regulon, coregulated with the expression of xynA1, conferring the ability for efficient assimilation and catabolism of the aldouronate product generated by a multimodular cell surface-anchored GH10 endoxylanase. This cluster offers a desirable metabolic potential for bacterial conversion of hemicellulose fractions of hardwood and crop residues to biobased products. PMID- 17921312 TI - H2O2 is required for optimal establishment of the Medicago sativa/Sinorhizobium meliloti symbiosis. AB - The symbiotic interaction between Medicago sativa and Sinorhizobium meliloti RmkatB(++) overexpressing the housekeeping catalase katB is delayed, and this delay is combined with an enlargement of infection threads. This result provides evidence that H(2)O(2) is required for optimal progression of infection threads through the root hairs and plant cell layers. PMID- 17921313 TI - Differential requirements for RAD51 in Physcomitrella patens and Arabidopsis thaliana development and DNA damage repair. AB - RAD51, the eukaryotic homolog of the bacterial RecA recombinase, plays a central role in homologous recombination (HR) in yeast and animals. Loss of RAD51 function causes lethality in vertebrates but not in other animals or in the flowering plant Arabidopsis thaliana, suggesting that RAD51 is vital for highly developed organisms but not for others. Here, we found that loss of RAD51 function in the moss Physcomitrella patens, a plant of less complexity, caused a significant vegetative phenotype, indicating an important function for RAD51 in this organism. Moreover, loss of RAD51 caused marked hypersensitivity to the double-strand break-inducing agent bleomycin in P. patens but not in Arabidopsis. Therefore, HR is used for somatic DNA damage repair in P. patens but not in Arabidopsis. These data imply fundamental differences in the use of recombination pathways between plants. Moreover, these data demonstrate that the importance of RAD51 for viability is independent of taxonomic position or complexity of an organism. The involvement of HR in DNA damage repair in the slowly evolving species P. patens but not in fast-evolving Arabidopsis suggests that the choice of the recombination pathway is related to the speed of evolution in plants. PMID- 17921314 TI - Polar growth in the infectious hyphae of the phytopathogen ustilago maydis depends on a virulence-specific cyclin. AB - The maize smut fungus Ustilago maydis switches from yeast to hyphal growth to infect maize (Zea mays) plants. This switching is promoted by mating of compatible cells and seems to be required for plant penetration. Although many genes distinctively expressed during this dimorphic switch have been identified and shown to be essential for the infection process, none seems to be explicitly required for polar growth control. Here, we report the characterization of pcl12, encoding a cyclin that interacts specifically with Cdk5, an essential cyclin dependent kinase with regulatory roles in morphogenesis in U. maydis. Pcl12 fulfills the requirements to be a virulence-specific regulator of polar growth in U. maydis. First, pcl12 expression is induced during the pathogenic development. Secondly, Pcl12 is sufficient to induce hyperpolarized growth in U. maydis cells, as haploid cells overexpressing pcl12 in axenic conditions produce filaments that were morphologically indistinguishable from those produced during the infection process. Finally, cells defective in pcl12 showed impaired polar growth during the formation of the b-dependent filament, the induction of the conjugation tubes, or the formation of a promycelium in spore germination. However, in spite of this pivotal role during morphogenesis, pcl12 mutants were virulent. We discuss the implications of these results for the role of polar growth during the infection process. PMID- 17921315 TI - Arabidopsis relatives of the human lysine-specific Demethylase1 repress the expression of FWA and FLOWERING LOCUS C and thus promote the floral transition. AB - The timing of the developmental transition to flowering is critical to reproductive success in plants. Here, we show that Arabidopsis thaliana homologs of human Lysine-Specific Demethylase1 (LSD1; a histone H3-Lys 4 demethylase) reduce the levels of histone H3-Lys 4 methylation in chromatin of the floral repressor FLOWERING LOCUS C (FLC) and the sporophytically silenced floral repressor FWA. Two of the homologs, LSD1-LIKE1 (LDL1) and LSD1-LIKE2 (LDL2), act in partial redundancy with FLOWERING LOCUS D (FLD; an additional homolog of LSD1) to repress FLC expression. However, LDL1 and LDL2 appear to act independently of FLD in the silencing of FWA, indicating that there is target gene specialization within this histone demethylase family. Loss of function of LDL1 and LDL2 affects DNA methylation on FWA, whereas FLC repression does not appear to involve DNA methylation; thus, members of the LDL family can participate in a range of silencing mechanisms. PMID- 17921316 TI - Arabidopsis cotyledon-specific chloroplast biogenesis factor CYO1 is a protein disulfide isomerase. AB - Chloroplast development in cotyledons differs in a number of ways from that in true leaves, but the cotyledon-specific program of chloroplast biogenesis has not been clarified. The cyo1 mutant in Arabidopsis thaliana has albino cotyledons but normal green true leaves. Chloroplasts develop abnormally in cyo1 mutant plants grown in the light, but etioplasts are normal in mutants grown in the dark. We isolated CYO1 by T-DNA tagging and verified that the mutant allele was responsible for the albino cotyledon phenotype by complementation. CYO1 has a C(4)-type zinc finger domain similar to that of Escherichia coli DnaJ. CYO1 is expressed mainly in young plants under light conditions, and the CYO1 protein localizes to the thylakoid membrane in chloroplasts. Transcription of nuclear photosynthetic genes is generally unaffected by the cyo1 mutation, but the level of photosynthetic proteins is decreased in cyo1 mutants. Recombinant CYO1 accelerates disulfide bond reduction in the model substrate insulin and renatures RNase A, indicating that CYO1 has protein disulfide isomerase activity. These results suggest that CYO1 has a chaperone-like activity required for thylakoid biogenesis in cotyledons. PMID- 17921317 TI - Two calcium-dependent protein kinases, CPK4 and CPK11, regulate abscisic acid signal transduction in Arabidopsis. AB - Many biochemical approaches show functions of calcium-dependent protein kinases (CDPKs) in abscisic acid (ABA) signal transduction, but molecular genetic evidence linking defined CDPK genes with ABA-regulated biological functions at the whole-plant level has been lacking. Here, we report that ABA stimulated two homologous CDPKs in Arabidopsis thaliana, CPK4 and CPK11. Loss-of-function mutations of CPK4 and CPK11 resulted in pleiotropic ABA-insensitive phenotypes in seed germination, seedling growth, and stomatal movement and led to salt insensitivity in seed germination and decreased tolerance of seedlings to salt stress. Double mutants of the two CDPK genes had stronger ABA- and salt responsive phenotypes than the single mutants. CPK4- or CPK11-overexpressing plants generally showed inverse ABA-related phenotypes relative to those of the loss-of-function mutants. Expression levels of many ABA-responsive genes were altered in the loss-of-function mutants and overexpression lines. The CPK4 and CPK11 kinases both phosphorylated two ABA-responsive transcription factors, ABF1 and ABF4, in vitro, suggesting that the two kinases may regulate ABA signaling through these transcription factors. These data provide in planta genetic evidence for the involvement of CDPK/calcium in ABA signaling at the whole-plant level and show that CPK4 and CPK11 are two important positive regulators in CDPK/calcium-mediated ABA signaling pathways. PMID- 17921318 TI - Human ribosomal protein L13a is dispensable for canonical ribosome function but indispensable for efficient rRNA methylation. AB - Previously, we demonstrated that treatment of monocytic cells with IFN-gamma causes release of ribosomal protein L13a from the 60S ribosome and subsequent translational silencing of Ceruloplasmin (Cp) mRNA. Here, evidence using cultured cells demonstrates that Cp mRNA silencing is dependent on L13a and that L13a deficient ribosomes are competent for global translational activity. Human monocytic U937 cells were stably transfected with two different shRNA sequences for L13a and clonally selected for more than 98% abrogation of total L13a expression. Metabolic labeling of these cells showed rescue of Cp translation from the IFN-gamma mediated translational silencing activity. Depletion of L13a caused significant reduction of methylation of ribosomal RNA and of cap independent translation mediated by Internal Ribosome Entry Site (IRES) elements derived from p27, p53, and SNAT2 mRNAs. However, no significant differences in the ribosomal RNA processing, polysome formation, global translational activity, translational fidelity, and cell proliferation were observed between L13a deficient and wild-type control cells. These results support the notion that ribosome can serve as a depot for releasable translation-regulatory factors unrelated to its basal polypeptide synthetic function. Unlike mammalian cells, the L13a homolog in yeast is indispensable for growth. Thus, L13a may have evolved from an essential ribosomal protein in lower eukaryotes to having a role as a dispensable extra-ribosomal function in higher eukaryotes. PMID- 17921320 TI - On substrate selection for ATP synthesis in the failing human myocardium. PMID- 17921319 TI - Impaired activation of ATP-sensitive K+ channels in endocardial myocytes from left ventricular hypertrophy. AB - ATP-sensitive K(+) (K(ATP)) channels are essential for maintaining the cellular homeostasis against metabolic stress. Myocardial remodeling in various pathologies may alter this adaptive response to such stress. It was reported that transmural electrophysiological heterogeneity exists in ventricular myocardium. Therefore, we hypothesized that the K(ATP) channel properties might be altered in hypertrophied myocytes from endocardium. To test this hypothesis, we determined the K(ATP) channel currents using the perforated patch-clamp technique, open cell attached patches, and excised inside-out patches in both endocardial and epicardial myocytes isolated from hypertrophied [spontaneous hypertensive rats (SHR)] vs. normal [Wistar-Kyoto rats (WKY)] left ventricle. In endocardial cells, K(ATP) channel currents (I(K,ATP)), produced by 2 mM CN(-) and no glucose at 0 mV, were significantly smaller (P < 0.01), and time required to reach peak currents after onset of K(ATP) channel opening (Time(onset to peak)) was significantly longer (319 +/- 46 vs. 177 +/- 37 s, P = 0.01) in the SHR group (n = 9) than the WKY group (n = 13). However, in epicardial cells, there were no differences in I(K,ATP) and Time(onset to peak) between the groups (SHR, n = 12; WKY, n = 12). The concentration-open probability-response curves obtained during the exposure of open cells and excised patches to exogenous ATP revealed the impaired K(ATP) channel activation in endocardial myocytes from SHR. In conclusion, K(ATP) channel activation under metabolic stress was impaired in endocardial cells from rat hypertrophied left ventricle. The deficit of endocardial K(ATP) channels to decreased intracellular ATP might contribute to the maladaptive response of hypertrophied hearts to ischemia. PMID- 17921321 TI - Nucleoside and nucleobase transporters of primary human cardiac microvascular endothelial cells: characterization of a novel nucleobase transporter. AB - Levels of cardiovascular active metabolites, like adenosine, are regulated by nucleoside transporters of endothelial cells. We characterized the nucleoside and nucleobase transport capabilities of primary human cardiac microvascular endothelial cells (hMVECs). hMVECs accumulated 2-[3H]chloroadenosine via the nitrobenzylmercaptopurine riboside-sensitive equilibrative nucleoside transporter 1 (ENT1) at a V(max) of 3.4 +/- 1 pmol.microl(-1).s(-1), with no contribution from the nitrobenzylmercaptopurine riboside-insensitive ENT2. Inhibition of 2 chloroadenosine uptake by ENT1 blockers produced monophasic inhibition curves, which are also compatible with minimal ENT2 expression. The nucleobase [3H]hypoxanthine was accumulated within hMVECs (K(m) = 96 +/- 37 microM; V(max) = 1.6 +/- 0.3 pmol.microl(-1).s(-1)) despite the lack of a known nucleobase transport system. This novel transporter was dipyridamole-insensitive but could be inhibited by adenine (K(i) = 19 +/- 7 microM) and other purine nucleobases, including chemotherapeutic analogs. A variety of other cell types also expressed the nucleobase transporter, including the nucleoside transporter-deficient PK(15) cell line (PK15NTD). Further characterization of [3H]hypoxanthine uptake in the PK15NTD cells showed no dependence on Na(+) or H(+). PK15NTD cells expressing human ENT2 accumulated 4.5-fold more [3H]hypoxanthine in the presence of the ENT2 inhibitor dipyridamole than did PK15NTD cells or hMVECs, suggesting trapping of ENT2-permeable metabolites. Understanding the nucleoside and nucleobase transporter profiles in the vasculature will allow for further study into their roles in pathophysiological conditions such as hypoxia or ischemia. PMID- 17921323 TI - Acetaminophen-mediated cardioprotection via inhibition of the mitochondrial permeability transition pore-induced apoptotic pathway. AB - Our laboratory has previously reported that acetaminophen confers functional cardioprotection following cardiac insult, including ischemia/reperfusion, hypoxia/reoxygenation, and exogenous peroxynitrite administration. In the present study, we further examined the mechanism of acetaminophen-mediated cardioprotection following ischemia/reperfusion injury. Langendorff-perfused guinea pig hearts were exposed to acute treatment with acetaminophen (0.35 mM) or vehicle beginning at 15 min of a 30-min baseline stabilization period. Low-flow global myocardial ischemia was subsequently induced for 30 min followed by 60 min of reperfusion. At the completion of reperfusion, hearts were homogenized and separated into cytosolic and mitochondrial fractions. Mitochondrial swelling and mitochondrial cytochromec release were assessed and found to be significantly and completely reduced in acetaminophen- vs. vehicle-treated hearts following reperfusion. In a separate group of hearts, ventricular myocytes were isolated and subjected to fluorescence-activated cell sorting. Acetaminophen-treated hearts showed a significant decrease in late stage apoptotic myocytes compared with vehicle-treated hearts following injury (58 +/- 1 vs. 81 +/- 5%, respectively). These data, together with electron micrograph analysis, suggest that acetaminophen mediates cardioprotection, in part, via inhibition of the mitochondrial permeability transition pore and subsequent apoptotic pathway. PMID- 17921322 TI - Interactions between oxidative stress and inflammation in salt-sensitive hypertension. AB - The goal of this study was to test the hypothesis that increases in oxidative stress in Dahl S rats on a high-salt diet help to stimulate renal nuclear factor kappaB (NF-kappaB), renal proinflammatory cytokines, and chemokines, thus contributing to hypertension, renal damage, and dysfunction. We specifically studied whether antioxidant treatment of Dahl S rats on high Na intake would decrease renal inflammation and thus attenuate the hypertensive and adverse renal responses. Sixty-four 7- to 8-wk-old Dahl S or R/Rapp strain rats were maintained for 5 wk on high Na (8%) or high Na + vitamins C (1 g/l in drinking water) and E (5,000 IU/kg in food). Arterial and venous catheters were implanted at day 21. By day 35 in the high-Na S rats, antioxidant treatment significantly increased the renal reduced-to-oxidized glutathione ratio and decreased renal cortical H(2)O(2) and O(2)(*-) release and renal NF-kappaB. Antioxidant treatment with vitamins C and E in high-Na S rats also decreased renal monocytes/macrophages in the glomeruli, cortex, and medulla, decreased tumor necrosis factor-alpha by 39%, and decreased monocyte chemoattractant protein-1 by 38%. Vitamin-treated, high-Na S rats also experienced decreases in arterial pressure, urinary protein excretion, renal tubulointerstitial damage, and glomerular necrosis and increases in glomerular filtration rate and renal plasma flow. In conclusion, antioxidant treatment of high-Na Dahl S rats decreased renal inflammatory cytokines and chemokines, renal immune cells, NF-kappaB, and arterial pressure and improved renal function and damage. PMID- 17921324 TI - IL-17 stimulates MMP-1 expression in primary human cardiac fibroblasts via p38 MAPK- and ERK1/2-dependent C/EBP-beta , NF-kappaB, and AP-1 activation. AB - Matrix metalloproteinases (MMPs) degrade collagen and mediate tissue remodeling. The novel cytokine IL-17 is expressed during various inflammatory conditions and modulates MMP expression. We investigated the effect of IL-17 on MMP-1 expression in primary human cardiac fibroblasts (HCF) and delineated the signaling pathways involved. HCF were treated with recombinant human IL-17. MMP-1 expression was analyzed by Northern blotting, RT-quantitative PCR, Western blotting, and ELISA; transcriptional induction and transcription factor binding by EMSA, ELISA, and reporter assay; and p38 MAPK and ERK1/2 activation by protein kinase assays and Western blotting. Signal transduction pathways were investigated using pharmacological inhibitors, small interfering RNA (siRNA), and adenoviral dominant-negative expression vectors. IL-17 stimulated MMP-1 gene transcription, net mRNA levels, protein, and promoter-reporter activity in HCF. This response was blocked by IL-17 receptor-Fc chimera and IL-17 receptor antibodies, but not by IL-6, TNF-alpha, or IL-1beta antibodies. IL-17-stimulated type I collagenase activity was inhibited by the MMP inhibitor GM-6001 and by siRNA-mediated MMP-1 knockdown. IL-17 stimulated activator protein-1 [AP-1 (c-Fos, c-Jun, and Fra-1)], NF-kappaB (p50 and p65), and CCAAT enhancer-binding protein (C/EBP)-beta DNA binding and reporter gene activities, effects attenuated by antisense oligonucleotides, siRNA-mediated knockdown, or expression of dominant-negative signaling proteins. Inhibition of AP-1, NF-kappaB, or C/EBP activation attenuated IL-17-stimulated MMP-1 expression. IL-17 induced p38 MAPK and ERK1/2 activation, and inhibition by SB-203580 and PD-98059 blunted IL-17-mediated transcription factor activation and MMP-1 expression. Our data indicate that IL-17 induces MMP 1 in human cardiac fibroblasts directly via p38 MAPK- and ERK-dependent AP-1, NF kappaB, and C/EBP-beta activation and suggest that IL-17 may play a critical role in myocardial remodeling. PMID- 17921326 TI - Exercise-mediated regulation of Hsp70 expression following aerobic exercise training. AB - An issue central to understanding the biological benefits associated with regular exercise training is to elucidate the intracellular mechanisms governing exercise conferred cardioprotection. Heat shock proteins (HSPs), most notably the inducible 70-kDa HSP family member Hsp70, are believed to participate in the protection of the myocardium during cardiovascular stress. Following acute exercise, activation of PKA mediates the suppression of an intermediary protein kinase, ERK1/2, which phosphorylates and suppresses the activation of the heat shock transcription factor 1 (HSF1). However, following exercise training, ERK1/2 has been reported to regulate the transcriptional activation of several genes involved in cell growth and proliferation and has been shown to be associated with training-mediated myocardial hypertrophy. The present project examined the transcriptional activation of hsp70 gene expression in acutely exercised (60 min at 30 m/min) naive sedentary and aerobically trained (8 wk, low intensity) male Sprague-Dawley rats. Following acute exercise stress, no significant differences were demonstrated in the expression of myocardial Hsp70 mRNA and activation of PKA between sedentary and trained animals. However, trained animals elicited expression of the hsp70 gene (P < 0.05) in the presence of elevated ERK1/2 activation. Given the association of ERK1/2 and the suppression of hsp70 gene expression following acute exercise in naive sedentary rats, these results suggest that training results in adaptations that allow for the simultaneous initiation of both proliferative and protective responses. While it is unclear what factors are associated with this training-related shift, increases in HSF1 DNA binding affinity (P < 0.05) and posttranscriptional modifications of the Hsp70 transcript are suggested. PMID- 17921325 TI - Impaired myocardial metabolic reserve and substrate selection flexibility during stress in patients with idiopathic dilated cardiomyopathy. AB - Under resting conditions, the failing heart shifts fuel use toward greater glucose and lower free fatty acid (FFA) oxidation. We hypothesized that chronic metabolic abnormalities in patients with dilated cardiomyopathy (DCM) are associated with the absence of the normal increase in myocardial glucose uptake and maintenance of cardiac mechanical efficiency in response to pacing stress. In 10 DCM patients and 6 control subjects, we measured coronary flow by intravascular ultrasonometry and sampled arterial and coronary sinus blood. Myocardial metabolism was determined at baseline, during atrial pacing at 130 beats/min, and at 15 min of recovery by infusion of [(3)H]oleate and [(13)C]lactate and measurement of transmyocardial arteriovenous differences of oxygen and metabolites. At baseline, DCM patients showed depressed coronary flow, reduced uptake and oxidation of FFA, and preferential utilization of carbohydrates. During pacing, glucose uptake increased by 106% in control subjects but did not change from baseline in DCM patients. Lactate release increased by 122% in DCM patients but not in control subjects. Cardiac mechanical efficiency in DCM patients was not different compared with control subjects at baseline but was 34% lower during stress. Fatty acid uptake and oxidation did not change with pacing in either group. Our results show that in DCM there is preferential utilization of carbohydrates, which is associated with reduced flow and oxygen consumption at rest and an impaired ability to increase glucose uptake during stress. These metabolic abnormalities might contribute to progressive cardiac deterioration and represent a target for therapeutic strategies aimed at modulating cardiac substrate utilization. PMID- 17921327 TI - Changes in forearm muscle temperature alter renal vascular responses to isometric handgrip. AB - The purpose of the present study was to examine the effect of heating and cooling the forearm muscles on renal vascular responses to ischemic isometric handgrip (IHG). It was hypothesized that heating and cooling the forearm would augment and attenuate, respectively, renal vascular responses to IHG. Renal vascular responses to IHG were studied during forearm heating at 39 degrees C (n = 15, 26 +/- 1 yr) and cooling at 26 degrees C (n = 12, 26 +/- 1 yr). For a control trial, subjects performed the experimental protocol while the forearm was normothermic (approximately 34 degrees C). Muscle temperature (measured by intramuscular probe) was controlled by changing the temperature of water cycling through a water-perfused sleeve. The experimental protocol was as follows: 3 min at baseline, 1 min of ischemia, ischemic IHG to fatigue, and 2 min of postexercise muscle ischemia. At rest, renal artery blood velocity (RBV; Doppler ultrasound) and renal vascular conductance (RVC = RBV/mean arterial blood pressure) were not different between normothermia and the two thermal conditions. During ischemic IHG, there were greater decreases in RBV and RVC in the heating trial. However, RBV and RVC were similar during postexercise muscle ischemia during heating and normothermia. RVC decreased less during cooling than in normothermia while the subjects performed the ischemic IHG protocol. During postexercise muscle ischemia, RVC was greater during cooling than in normothermia. These results indicate that heating augments mechanoreceptor-mediated renal vasoconstriction whereas cooling blunts metaboreceptor-mediated renal vasoconstriction. PMID- 17921328 TI - Protective roles of adenosine A1, A2A, and A3 receptors in skeletal muscle ischemia and reperfusion injury. AB - Although adenosine exerts cardio-and vasculoprotective effects, the roles and signaling mechanisms of different adenosine receptors in mediating skeletal muscle protection are not well understood. We used a mouse hindlimb ischemia reperfusion model to delineate the function of three adenosine receptor subtypes. Adenosine A(3) receptor-selective agonist 2-chloro-N(6)-(3-iodobenzyl)adenosine 5'-N-methyluronamide (Cl-IBMECA; 0.07 mg/kg ip) reduced skeletal muscle injury with a significant decrease in both Evans blue dye staining (5.4 +/- 2.6%, n = 8 mice vs. vehicle-treated 28 +/- 6%, n = 7 mice, P < 0.05) and serum creatine kinase level (1,840 +/- 910 U/l, n = 13 vs. vehicle-treated 12,600 +/- 3,300 U/l, n = 14, P < 0.05), an effect that was selectively blocked by an A(3) receptor antagonist 3-ethyl-5-benzyl-2-methyl-6-phenyl-4-phenylethynyl-1,4-(+/-) dihydropyridine-3,5-dicarboxylate (MRS-1191; 0.05 mg/kg). The adenosine A(1) receptor agonist 2-chloro-N(6)-cyclopentyladenosine (CCPA; 0.05 mg/kg) also exerted a cytoprotective effect, which was selectively blocked by the A(1) antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX; 0.2 mg/kg). The adenosine A(2A) receptor agonist 2-p-(2-carboxyethyl)phenethylamino-5'-N ethylcarboxamidoadenosine (CGS-21680; 0.07 mg/kg)-induced decrease in skeletal muscle injury was selectively blocked by the A(2A) antagonist 2-(2-furanyl)-7-[3 (4-methoxyphenyl)propyl]-7H-pyrazolo[4,3-e] [1,2,4]triazolo[1,5-C]pyrimidin-5 amine (SCH-442416; 0.017 mg/kg). The protection induced by the A(3) receptor was abrogated in phospholipase C-beta2/beta3 null mice, but the protection mediated by the A(1) or A(2A) receptor remained unaffected in these animals. The adenosine A(3) receptor is a novel cytoprotective receptor that signals selectively via phospholipase C-beta and represents a new target for ameliorating skeletal muscle injury. PMID- 17921329 TI - Involvement of COX-1 in A3 adenosine receptor-mediated contraction through endothelium in mice aorta. AB - We investigated whether A(3) adenosine receptor (A(3)AR) is involved in endothelium-mediated contraction through cyclooxygenases (COXs) with the use of wild-type (WT) and A(3) knockout (A(3)KO) mice aorta. A(3)AR-selective agonist, Cl-IBMECA, produced a concentration-dependent contraction (EC(50): 2.9 +/- 0.2 x 10(-9) M) in WT mouse aorta with intact endothelium (+E) and negligible effects in A(3)KO +E aorta. At 10(-7) M, contractions produced by Cl-IBMECA were 29% in WT +E, while being insignificant in A(3)KO +E aorta. Cl-IBMECA-induced responses were abolished in endothelium-denuded tissues (-E), in both WT and A(3)KO aorta. A(3)AR gene and protein expression were reduced by 74 and 72% (P < 0.05), respectively, in WT -E compared with WT +E aorta, while being undetected in A(3)KO +E/-E aorta. Indomethacin (nonspecific COXs blocker, 10(-5) M), SC-560 (specific COX-1 blocker, 10(-8) M), SQ 29549 (thromboxane prostanoid receptor antagonist, 10(-6) M), and furegrelate (thromboxane synthase inhibitor, 10(-5) M) inhibited Cl-IBMECA-induced contraction significantly. Cl-IBMECA-induced thromboxane B(2) production was also attenuated significantly by indomethacin, SC 560, and furegrelate in WT +E aorta, while having negligible effects in A(3)KO +E aorta. NS-398 (specific COX-2 blocker) produced negligible inhibition of Cl IBMECA-induced contraction in both WT +E and A(3)KO +E aorta. Cl-IBMECA-induced increase in COX-1 and thromboxane prostanoid receptor expression were significantly inhibited by MRS1523, a specific A(3)AR antagonist in WT +E aorta. Expression of both A(3)AR and COX-1 was located mostly on endothelium of WT and A(3)KO +E aorta. These results demonstrate for the first time the involvement of COX-1 pathway in A(3)AR-mediated contraction via endothelium. PMID- 17921330 TI - Increased metaboreflex activity is related to exercise intolerance in heart transplant patients. AB - Heart transplantation does not normalize exercise capacity or the ventilatory response to exercise. We hypothesized that excessive muscle reflex activity, as assessed by the muscle sympathetic nerve activity (MSNA) response to handgrip exercise, persists after cardiac transplantation and that this mechanism is related to exercise hyperpnea in heart transplant recipients (HTRs). We determined the MSNA, ventilatory, and cardiovascular responses to isometric and dynamic handgrips in 11 HTRs and 10 matched control subjects. Handgrips were followed by a post-handgrip ischemia to isolate the metaboreflex contribution to exercise responses. HTRs and control subjects also underwent recordings during isocapnic hypoxia and a maximal, symptom-limited, cycle ergometer exercise test. HTRs had higher resting MSNA (P < 0.01) and heart rate (P < 0.01) than the control subjects. Isometric handgrip increased MSNA in HTRs more than in the controls (P = 0.003). Dynamic handgrip increased MSNA only in HTRs. During post handgrip ischemia, MSNA and ventilation remained more elevated in HTRs (P < 0.05). The MSNA and ventilatory responses to hypoxia were also higher in HTRs (both P < 0.04). In HTRs, metaboreflex overactivity was related to the ventilatory response to exercise, characterized by the regression slope relating ventilation to CO(2) output (r = +0.8; P < 0.05) and a lower peak ventilation (r = +0.81; P < 0.05) during cycle ergometer exercise tests. However, increased chemoreflex sensitivity (r = +0.91; P < 0.005), but not metaboreflex activity, accounted for the lower peak ventilation during exercise in a stepwise regression analysis. In conclusion, heart transplantation does not normalize muscle metaboreceptor activity; both increased metaboreflex and chemoreflex control are related to exercise intolerance in HTRs. PMID- 17921331 TI - Effects of cyclooxygenase inhibition on canine coronary artery blood flow and thrombosis. AB - This study was designed to determine the effect of inhibitors of cyclooxygenase (COX)-1, COX-2, and the nonselective COX inhibitor naproxen on coronary vasoactivity and thrombogenicity under baseline and lipopolysaccharide (LPS) induced inflammatory conditions. We hypothesize that endothelial COX-1 is the primary COX isoform in the canine normal coronary artery, which mediates arachidonic acid (AA)-induced vasodilatation. However, COX-2 can be induced and overexpressed by inflammatory mediators and becomes the major local COX isoform responsible for the production of antithrombotic prostaglandins during systemic inflammation. The interventions included the selective COX-1 inhibitor SC-560 (0.3 mg/kg iv), the selective COX-2 inhibitor nimesulide (5 mg/kg iv), or the nonselective COX inhibitor naproxen (3 mg/kg iv). The selective prostacyclin (IP) receptor antagonist RO-3244794 (RO) was used as an investigational tool to delineate the role of prostacyclin (PGI(2)) in modulating vascular reactivity. AA induced vasodilatation of the left circumflex coronary artery was suppressed to a similar extent by each of the COX inhibitors and RO. The data suggest that AA induced vasodilatation in the normal coronary artery is mediated by a single COX isoform, the constitutive endothelial COX-1, which is reported to be susceptible to COX-2 inhibitors. The effect of the COX inhibitors on thrombus formation was evaluated in a model of carotid artery thrombosis secondary to electrolytic induced vessel wall injury. Pretreatment with LPS (0.5 mg/kg iv) induced a systemic inflammatory response and prolonged the time-to-occlusive thrombus formation, which was reduced in the LPS-treated animals by the administration of nimesulide. In contrast, neither SC-560 nor naproxen influenced the time to thrombosis in the animals pretreated with LPS. The data are of significance in view of reported adverse cardiovascular events observed in clinical trials involving the use of selective COX-2 inhibitors, thereby suggesting that the endothelial constitutive COX-1 and the inducible vascular COX-2 serve important functions in maintaining vascular homeostasis. PMID- 17921333 TI - Interaction of BMP10 with Tcap may modulate the course of hypertensive cardiac hypertrophy. AB - Elevated wall stress by hypertension induces an adaptive myocardial hypertrophy via releasing prohypertrophic hormones such as angiotensin II. In this study, we investigated the involvement of bone morphogenetic protein-10 (BMP10) in hypertension-induced cardiac hypertrophy. Expression of BMP10 was increased in the hypertrophied ventricles from hypertensive rats. BMP10 localized on cell surface and at stretch-sensing Z disc of cardiomyocytes, where BMP10 interacted with a protein called titin-cap (Tcap). A rare variant of the human BMP10 gene, Thr326Ile, was found to be associated with hypertensive dilated cardiomyopathy. The variant BMP10 demonstrated decreased binding to Tcap and increased extracellular secretion. Conditioned medium from cells transfected with wild-type or variant BMP10 induced hypertrophy in rat neonatal cardiomyocytes, except that medium from variant BMP10-carrying cells showed an enhanced effect reflecting the increased secretion. These observations suggested that hypertension induced expression of prohypertrophic BMP10, and the hypertrophic effect of BMP10 was modulated, at least in part, by its binding to Tcap at the Z disc. PMID- 17921332 TI - Inducible and myocyte-specific inhibition of PKCalpha enhances cardiac contractility and protects against infarction-induced heart failure. AB - Mice null for the gene encoding protein kinase Calpha (Prkca), or mice treated with pharmacologic inhibitors of the PKCalpha/beta/gamma isoforms, show an augmentation in cardiac contractility that appears to be cardioprotective. However, it remains uncertain if PKCalpha itself functions in a myocyte autonomous manner to affect cardioprotection in vivo. Here we generated cardiac myocyte-specific transgenic mice using a tetracycline-inducible system to permit controlled expression of dominant negative PKCalpha in the heart. Consistent with the proposed function of PKCalpha, induction of dominant negative PKCalpha expression in the adult heart enhanced baseline cardiac contractility. This increase in cardiac contractility was associated with a partial protection from long-term decompensation and secondary dilated cardiomyopathy after myocardial infarction injury. Similarly, Prkca null mice were also partially protected from infarction-induced heart failure, although the area of infarction injury was identical to controls. Thus, myocyte autonomous inhibition of PKCalpha protects the adult heart from decompensation and dilated cardiomyopathy after infarction injury in association with a primary enhancement in contractility. PMID- 17921334 TI - Isometric contraction induces rapid myocyte remodeling in cultured rat right ventricular papillary muscles. AB - The hypothesis that elevated systolic stress induces myocyte thickening has been difficult to test directly. We tested this hypothesis in working rat right ventricular papillary muscles using a recently developed technique for long-term muscle culture. Muscles were cultured for 36 h either isometrically at different levels of systolic stress or at physiological amounts and rates of shortening. Isometric contraction induced rapid increases in myocyte diameter regardless of the level of systolic stress, whereas control myocyte dimensions were maintained if physiological amounts and rates of systolic shortening were imposed. Myocyte thickening was accompanied by a significant decrease in cell length and number of sarcomeres in series along the long axis of the myocyte, suggesting that thickening may have occurred in part by rearrangement of existing sarcomeres. We conclude that the pattern of systolic shortening and/or diastolic lengthening regulates myocyte shape in working rat right ventricular papillary muscles, whereas systolic stress plays little or no role. PMID- 17921335 TI - Mechanisms of acrolein-induced myocardial dysfunction: implications for environmental and endogenous aldehyde exposure. AB - Aldehydes are ubiquitous pollutants generated during the combustion of organic materials and are present in air, water, and food. Several aldehydes are also endogenous products of lipid peroxidation and by-products of drug metabolism. Despite well-documented high reactivity of unsaturated aldehydes, little is known regarding their cardiovascular effects and their role in cardiac pathology. Accordingly, we examined the myocardial effects of the model unsaturated aldehyde acrolein. In closed-chest mice, intravenous acrolein (0.5 mg/kg) induced rapid but reversible left ventricular dilatation and dysfunction. In mouse myocytes, micromolar acrolein acutely depressed myofilament Ca(2+) responsiveness without altering catecholamine sensitivity, similar to the phenotype of stunned myocardium. Immunoblotting revealed increased acrolein-protein adducts and protein-carbonyls in both acrolein-exposed myocardium (1.8-fold increase, P < 0.002) and myocytes (6.4-fold increase, P < 0.02). Both the contractile dysfunction and adduct formation were markedly attenuated by pretreatment with the thiol donor N-acetylcysteine (5 mM). Two-dimensional gel electrophoresis and mass-assisted laser desorption/ionization time-of-flight mass spectrometry analysis revealed two groups of adducted proteins, sarcomeric/cytoskeletal proteins (cardiac alpha-actin, desmin, myosin light polypeptide 3) and energy metabolism proteins (mitochondrial creatine kinase-2, ATP synthase), indicating site-specific protein modification that was confirmed by immunohistochemical colocalization. We conclude that direct exposure to acrolein induces selective myofilament impairment, which may be, in part, related to the modification of proteins involved in myocardial contraction and energy metabolism. Myocardial dysfunction induced by acrolein and related aldehydes may be symptomatic of toxicological states associated with ambient or occupational exposures or drug toxicity. Moreover, aldehydes such as acrolein may mediate cardiac dysfunction in pathologies characterized by high-oxidative stress. PMID- 17921336 TI - Genome-wide linkage scan for submaximal exercise heart rate in the HERITAGE family study. AB - The purpose of this study was to identify regions of the human genome linked to submaximal exercise heart rates in the sedentary state and in response to a standardized 20-wk endurance training program in blacks and whites of the HERITAGE Family Study. A total of 701 polymorphic markers covering the 22 autosomes were used in the genome-wide linkage scan, with 328 sibling pairs from 99 white nuclear families and 102 pairs from 115 black family units. Steady-state heart rates were measured at the relative intensity of 60% maximal oxygen uptake (HR60) and at the absolute intensity of 50 W (HR50). Baseline phenotypes were adjusted for age, sex, and baseline body mass index (BMI) and training responses (posttraining minus baseline, Delta) were adjusted for age, sex, baseline BMI, and baseline value of the phenotype. Two analytic strategies were used, a multipoint variance components and a regression-based multipoint linkage analysis. In whites, promising linkages (LOD > 1.75) were identified on 18q21-q22 for baseline HR50 (LOD = 2.64; P = 0.0002) and DeltaHR60 (LOD = 2.10; P = 0.0009) and on chromosome 2q33.3 for DeltaHR50 (LOD = 2.13; P = 0.0009). In blacks, evidence of promising linkage for baseline HR50 was detected with several markers within the chromosomal region 10q24-q25.3 (peak LOD = 2.43, P = 0.0004 with D10S597). The most promising regions for fine mapping in the HERITAGE Family Study were found on 2q33 for HR50 training response in whites, on 10q25-26 for baseline HR60 in blacks, and on 18q21-22 for both baseline HR50 and DeltaHR60 in whites. PMID- 17921337 TI - Stable recombinase-mediated cassette exchange in Arabidopsis using Agrobacterium tumefaciens. AB - Site-specific integration is an attractive method for the improvement of current transformation technologies aimed at the production of stable transgenic plants. Here, we present a Cre-based targeting strategy in Arabidopsis (Arabidopsis thaliana) using recombinase-mediated cassette exchange (RMCE) of transferred DNA (T-DNA) delivered by Agrobacterium tumefaciens. The rationale for effective RMCE is the precise exchange of a genomic and a replacement cassette both flanked by two heterospecific lox sites that are incompatible with each other to prevent unwanted cassette deletion. We designed a strategy in which the coding region of a loxP/lox5171-flanked bialaphos resistance (bar) gene is exchanged for a loxP/lox5171-flanked T-DNA replacement cassette containing the neomycin phosphotransferase (nptII) coding region via loxP/loxP and lox5171/lox5171 directed recombination. The bar gene is driven by the strong 35S promoter, which is located outside the target cassette. This placement ensures preferential selection of RMCE events and not random integration events by expression of nptII from this same promoter. Using root transformation, during which Cre was provided on a cotransformed T-DNA, 50 kanamycin-resistant calli were selected. Forty-four percent contained a correctly exchanged cassette based on PCR analysis, indicating the stringency of the selection system. This was confirmed for the offspring of five analyzed events by Southern-blot analysis. In four of the five analyzed RMCE events, there were no additional T-DNA insertions or they easily segregated, resulting in high-efficiency single-copy RMCE events. Our approach enables simple and efficient selection of targeting events using the advantages of Agrobacterium-mediated transformation. PMID- 17921338 TI - Role of the PsbI protein in photosystem II assembly and repair in the cyanobacterium Synechocystis sp. PCC 6803. AB - The involvement of the PsbI protein in the assembly and repair of the photosystem II (PSII) complex has been studied in the cyanobacterium Synechocystis sp. PCC 6803. Analysis of PSII complexes in the wild-type strain showed that the PsbI protein was present in dimeric and monomeric core complexes, core complexes lacking CP43, and in reaction center complexes containing D1, D2, and cytochrome b-559. In addition, immunoprecipitation experiments and the use of a histidine tagged derivative of PsbI have revealed the presence in the thylakoid membrane of assembly complexes containing PsbI and either the precursor or mature forms of D1. Analysis of PSII assembly in the psbI deletion mutant and in strains lacking PsbI together with other PSII subunits showed that PsbI was not required for formation of PSII reaction center complexes or core complexes, although levels of unassembled D1 were reduced in its absence. However, loss of PsbI led to a dramatic destabilization of CP43 binding within monomeric and dimeric PSII core complexes. Despite the close structural relationship between D1 and PsbI in the PSII complex, PsbI turned over much slower than D1, whereas high light-induced turnover of D1 was accelerated in the absence of PsbI. Overall, our results suggest that PsbI is an early assembly partner for D1 and that it plays a functional role in stabilizing the binding of CP43 in the PSII holoenzyme. PMID- 17921339 TI - Digalactosyldiacylglycerol is required for stabilization of the oxygen-evolving complex in photosystem II. AB - The galactolipid digalactosyldiacylglycerol (DGDG) is present in the thylakoid membranes of oxygenic photosynthetic organisms such as higher plants and cyanobacteria. Recent x-ray crystallographic analysis of protein-cofactor supercomplexes in thylakoid membranes revealed that DGDG molecules are present in the photosystem II (PSII) complex (four molecules per monomer), suggesting that DGDG molecules play important roles in folding and assembly of subunits in the PSII complex. However, the specific role of DGDG in PSII has not been fully clarified. In this study, we identified the dgdA gene (slr1508, a ycf82 homolog) of Synechocystis sp. PCC6803 that presumably encodes a DGDG synthase involved in the biosynthesis of DGDG by comparison of genomic sequence data. Disruption of the dgdA gene resulted in a mutant defective in DGDG synthesis. Despite the lack of DGDG, the mutant cells grew as rapidly as the wild-type cells, indicating that DGDG is not essential for growth in Synechocystis. However, we found that oxygen evolving activity of PSII was significantly decreased in the mutant. Analyses of the PSII complex purified from the mutant cells indicated that the extrinsic proteins PsbU, PsbV, and PsbO, which stabilize the oxygen-evolving complex, were substantially dissociated from the PSII complex. In addition, we found that heat susceptibility but not dark-induced inactivation of oxygen-evolving activity was notably increased in the mutant cells in comparison to the wild-type cells, suggesting that the PsbU subunit is dissociated from the PSII complex even in vivo. These results demonstrate that DGDG plays important roles in PSII through the binding of extrinsic proteins required for stabilization of the oxygen evolving complex. PMID- 17921340 TI - Silencing of the mitochondrial ascorbate synthesizing enzyme L-galactono-1,4 lactone dehydrogenase affects plant and fruit development in tomato. AB - L-Galactono-1,4-lactone dehydrogenase (EC 1.3.2.3) catalyzes the last step in the main pathway of vitamin C (L-ascorbic acid) biosynthesis in higher plants. In this study, we first characterized the spatial and temporal expression of SlGalLDH in several organs of tomato (Solanum lycopersicum) plants in parallel with the ascorbate content. P(35S):Slgalldh(RNAi) silenced transgenic tomato lines were then generated using an RNAi strategy to evaluate the effect of any resulting modification of the ascorbate pool on plant and fruit development. In all P(35S):Slgalldh(RNAi) plants with reduced SlGalLDH transcript and activity, plant growth rate was decreased. Plants displaying the most severe effects (dwarf plants with no fruit) were excluded from further analysis. The most affected lines studied exhibited up to an 80% reduction in SlGalLDH activity and showed a strong reduction in leaf and fruit size, mainly as a consequence of reduced cell expansion. This was accompanied by significant changes in mitochondrial function and altered ascorbate redox state despite the fact that the total ascorbate content remained unchanged. By using a combination of transcriptomic and metabolomic approaches, we further demonstrated that several primary, like the tricarboxylic acid cycle, as well as secondary metabolic pathways related to stress response were modified in leaves and fruit of P(35S):Slgalldh(RNAi) plants. When taken together, this work confirms the complexity of ascorbate regulation and its link with plant metabolism. Moreover, it strongly suggests that, in addition to ascorbate synthesis, GalLDH could play an important role in the regulation of cell growth-related processes in plants. PMID- 17921341 TI - A BELL1-like gene of potato is light activated and wound inducible. AB - BELL1-like transcription factors interact with their protein partners from the KNOTTED1 family to bind to target genes and regulate numerous developmental and metabolic processes. In potato (Solanum tuberosum), the BELL1 transcription factor StBEL5 and its protein partner POTH1 regulate tuber formation by affecting hormone levels. Overexpression of StBEL5 in transgenic lines produces plants that consistently exhibit enhanced tuber formation, and the mRNA of this gene moves through phloem cells in a long-distance signaling pathway regulated by photoperiod. Whereas photoperiod mediates the movement of StBEL5 RNA, activation of transcription of the StBEL5 gene in leaves is regulated by white light, regardless of photoperiod or light intensity. Illumination with either red or blue light induces the StBEL5 promoter, whereas far-red light had no effect. As expected, the StBEL5 promoter harbors numerous conventional light-responsive cis acting elements like GT1, GATA, and AT1 motifs. Deletion constructs were analyzed to determine what sequences are involved in light activation. Transcriptional activity was also mediated by wounding on stems, insect predation on leaves, and photoperiod in stolons. These results demonstrate that StBEL5 gene activity in the leaf is correlated with wavelengths optimal for photosynthesis. The number of factors that affect the StBEL5 promoter supports the premise that the BELL1-like genes play a role in a wide range of functions. PMID- 17921342 TI - Development and application of novel constructs to score C:G-to-T:A transitions and homologous recombination in Arabidopsis. AB - We report on the development of five missense mutants and one recombination substrate of the beta-glucuronidase (GUS)-encoding gene of Escherichia coli and their use for detecting mutation and recombination events in transgenic Arabidopsis (Arabidopsis thaliana) plants by reactivation of GUS activity in clonal sectors. The missense mutants were designed to find C:G-to-T:A transitions in a symmetrical sequence context and are in that respect complementary to previously published GUS point mutants. Small peptide tags (hemagglutinin tag and Strep tag II) and green fluorescent protein were translationally fused to GUS, which offers possibilities to check for mutant GUS production levels. We show that spontaneous mutation and recombination events took place. Mutagenic treatment of the plants with ethyl methanesulfonate and ultraviolet-C increased the number of mutations, validating the use of these constructs to measure mutation and recombination frequencies in plants exposed to biotic or abiotic stress conditions, or in response to different genetic backgrounds. Plants were also subjected to heavy metals, methyl jasmonate, salicylic acid, and heat stress, for which no effect could be seen. Together with an ethyl methanesulfonate mutation induction level much higher than previously described, the need is illustrated for many available scoring systems in parallel. Because all GUS missense mutants were cloned in a bacterial expression vector, they can also be used to score mutation events in E. coli. PMID- 17921343 TI - A trafficking pathway for anthocyanins overlaps with the endoplasmic reticulum-to vacuole protein-sorting route in Arabidopsis and contributes to the formation of vacuolar inclusions. AB - Plants produce a very large number of specialized compounds that must be transported from their site of synthesis to the sites of storage or disposal. Anthocyanin accumulation has provided a powerful system to elucidate the molecular and cellular mechanisms associated with the intracellular trafficking of phytochemicals. Benefiting from the unique fluorescent properties of anthocyanins, we show here that in Arabidopsis (Arabidopsis thaliana), one route for anthocyanin transport to the vacuole involves vesicle-like structures shared with components of the secretory pathway. By colocalizing the red fluorescence of the anthocyanins with green fluorescent protein markers of the endomembrane system in Arabidopsis seedlings, we show that anthocyanins are also sequestered to the endoplasmic reticulum and to endoplasmic reticulum-derived vesicle-like structures targeted directly to the protein storage vacuole in a Golgi independent manner. Moreover, our results indicate that vacuolar accumulation of anthocyanins does not depend solely on glutathione S-transferase activity or ATP dependent transport mechanisms. Indeed, we observed a dramatic increase of anthocyanin-filled subvacuolar structures, without a significant effect on total anthocyanin levels, when we inhibited glutathione S-transferase activity, or the ATP-dependent transporters with vanadate, a general ATPase inhibitor. Taken together, these results provide evidence for an alternative novel mechanism of vesicular transport and vacuolar sequestration of anthocyanins in Arabidopsis. PMID- 17921344 TI - Ethylene and not embolism is required for wound-induced tylose development in stems of grapevines. AB - The pruning of actively growing grapevines (Vitis vinifera) resulted in xylem vessel embolisms and a stimulation of tylose formation in the vessels below the pruning wound. Pruning was also followed by a 10-fold increase in the concentration of ethylene at the cut surface. When the pruning cut was made under water and maintained in water, embolisms were prevented, but there was no reduction in the formation of tyloses or the accumulation of ethylene. Treatment of the stems with inhibitors of ethylene biosynthesis (aminoethoxyvinylglycine) and/or action (silver thiosulfate) delayed and greatly reduced the formation of tyloses in xylem tissue and the size and number of those that formed in individual vessels. Our data are consistent with the hypotheses that wound ethylene production is the cause of tylose formation and that embolisms in vessels are not directly required for wound-induced tylosis in pruned grapevines. The possible role of ethylene in the formation of tyloses in response to other stresses and during development, maturation, and senescence is discussed. PMID- 17921345 TI - Enhanced nitric oxide-mediated chemoreceptor inhibition and altered cyclic GMP signaling in rat carotid body following chronic hypoxia. AB - Multiple studies have shown that chronic hypoxia (CH) elicits a time-dependent upregulation of carotid body chemoreceptor sensitivity in mammals. In the present study, we demonstrate that enhanced excitation is accompanied by a parallel increase of nitric oxide (NO)-dependent inhibition, which acts via a CH-induced modification of the normal mechanism in O(2)-sensitive type I cells. The NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), elicits a progressively larger increase in carotid sinus nerve (CSN) chemoreceptor activity following incremental increases in CH exposure lasting 1-16 days. The inhibitory effect of the NO donor, S-nitroso-N-acetyl-penicillamine (SNAP), on CSN activity is enhanced following CH. However, the activation of soluble guanylate cyclase (sGC) by SNAP, assessed via production of cGMP, is impaired, along with decreased expression of sGC mRNA transcript. Inhibition of hypoxia-evoked Ca(2+) responses by SNAP is mediated via a cGMP/protein kinase G (PKG)-dependent mechanism in normal type I cells that is sensitive to the PKG inhibitor KT-5823, but following CH, inhibitory responses are minimally sensitive to PKG inhibition. The data are consistent with the hypothesis that CH hampers cGMP-mediated inhibition of type I cells in favor of an alternative mechanism. PMID- 17921346 TI - IL-21 mediated Foxp3 suppression leads to enhanced generation of antigen-specific CD8+ cytotoxic T lymphocytes. AB - Efforts to reproducibly isolate tumor antigen-specific T cells from patients would be facilitated by removing immunoregulatory barriers. Using a human model for eliciting T-cell responses to tumor-associated antigens, we develop a novel strategy that eliminates nearly all Foxp3-expressing cells through the combination of CD25 depletion and IL-21 treatment resulting in a more than 150 fold decrease in Foxp3(+) cells to virtually undetectable levels and a more than 200-fold increase in antigen-specific cytotoxic T lymphocytes (CTLs). The extent of Foxp3 elimination and degree of expansion of antigen-specific CTLs shown in this study have not previously been achievable and are unique to IL-21. We demonstrate for the first time a possible mechanism for IL-21-mediated expansion of antigen-specific CTLs that involves suppression of Foxp3-expressing cells and reversal of inhibition to tumor-associated antigen-specific CTL generation in vitro. Taken together, the combination of CD25 depletion and IL-21 exposure, by releasing regulatory constraints, leads to markedly enhanced CTL induction and represents a robust strategy for the ex vivo generation of antigen-specific T cells for adoptive cellular therapy. PMID- 17921347 TI - Locus control region elements HS1 and HS4 enhance the therapeutic efficacy of globin gene transfer in beta-thalassemic mice. AB - Globin gene transfer in autologous hematopoietic stem cells is a promising therapeutic option for subjects with beta-thalassemia major. In this approach, high level, erythroid-specific globin transgene expression should correct ineffective erythropoiesis and hemolytic anemia following the delivery of only 1 to 2 vector copies per cell. The generation of vectors that provide high-level globin expression and require low vector copy (VC) integration is therefore essential for both safety and efficacy. We show here the major roles played by 2 lesser-known locus control region elements, termed HS1 and HS4. Partial deletions within HS4 markedly reduce in vivo globin expression requiring multiple VC per cell to correct the anemia. Most strikingly, addition of HS1 to HS2-3-4 increases globin expression by 52%, yielding 9 g Hb/VC in beta-thalassemic mice. Thus, while vectors encoding HS2-3-4 provide curative levels of hemoglobin at 1 to 2 copies per cell, adding HS1 is a promising alternative strategy if upcoming clinical trials prove higher levels of expression to be necessary. PMID- 17921348 TI - Unexpected link between metal ion deficiency and autophagy in Aspergillus fumigatus. AB - Autophagy is the major cellular pathway for bulk degradation of cytosolic material and is required to maintain viability under starvation conditions. To determine the contribution of autophagy to starvation stress responses in the filamentous fungus Aspergillus fumigatus, we disrupted the A. fumigatus atg1 gene, encoding a serine/threonine kinase required for autophagy. The DeltaAfatg1 mutant showed abnormal conidiophore development and reduced conidiation, but the defect could be bypassed by increasing the nitrogen content of the medium. When transferred to starvation medium, wild-type hyphae were able to undergo a limited amount of growth, resulting in radial expansion of the colony. In contrast, the DeltaAfatg1 mutant was unable to grow under these conditions. However, supplementation of the medium with metal ions rescued the ability of the DeltaAfatg1 mutant to grow in the absence of a carbon or nitrogen source. Depleting the medium of cations by using EDTA was sufficient to induce autophagy in wild-type A. fumigatus, even in the presence of abundant carbon and nitrogen, and the DeltaAfatg1 mutant was severely growth impaired under these conditions. These findings establish a role for autophagy in the recycling of internal nitrogen sources to support conidiophore development and suggest that autophagy also contributes to the recycling of essential metal ions to sustain hyphal growth when exogenous nutrients are scarce. PMID- 17921349 TI - The Aspergillus fumigatus transcriptional regulator AfYap1 represents the major regulator for defense against reactive oxygen intermediates but is dispensable for pathogenicity in an intranasal mouse infection model. AB - Macrophages and neutrophils kill the airborne fungal pathogen Aspergillus fumigatus. The dependency of this killing process on reactive oxygen intermediates (ROI) has been strongly suggested. Therefore, we investigated the enzymatic ROI detoxifying system by proteome analysis of A. fumigatus challenged by H(2)O(2). Since many of the identified proteins and genes are apparently regulated by a putative Saccharomyces cerevisiae Yap1 homolog, the corresponding gene of A. fumigatus was identified and designated Afyap1. Nuclear localization of a functional AfYap1-eGFP fusion was stress dependent. Deletion of the Afyap1 gene led to drastically increased sensitivity of the deletion mutant against H(2)O(2) and menadione, but not against diamide and NO radicals. Proteome analysis of the DeltaAfyap1 mutant strain challenged with 2 mM H(2)O(2) indicated that 29 proteins are controlled directly or indirectly by AfYap1, including catalase 2. Despite its importance for defense against reactive agents, the Afyap1 deletion mutant did not show attenuated virulence in a murine model of Aspergillus infection. These data challenge the hypothesis that ROI such as superoxide anions and peroxides play a direct role in killing of A. fumigatus in an immunocompromised host. This conclusion was further supported by the finding that killing of A. fumigatus wild-type and DeltaAfyap1 mutant germlings by human neutrophilic granulocytes worked equally well irrespective of whether the ROI scavenger glutathione or an NADPH-oxidase inhibitor was added to the cells. PMID- 17921350 TI - Expression and characterization of the flocculin Flo11/Muc1, a Saccharomyces cerevisiae mannoprotein with homotypic properties of adhesion. AB - The Flo11/Muc1 flocculin has diverse phenotypic effects. Saccharomyces cerevisiae cells of strain background Sigma1278b require Flo11p to form pseudohyphae, invade agar, adhere to plastic, and develop biofilms, but they do not flocculate. We show that S. cerevisiae var. diastaticus strains, on the other hand, exhibit Flo11-dependent flocculation and biofilm formation but do not invade agar or form pseudohyphae. In order to study the nature of the Flo11p proteins produced by these two types of strains, we examined secreted Flo11p, encoded by a plasmid borne gene, in which the glycosylphosphatidylinositol anchor sequences had been replaced by a histidine tag. A protein of approximately 196 kDa was secreted from both strains, which upon purification and concentration, aggregated into a form with a very high molecular mass. When secreted Flo11p was covalently attached to microscopic beads, it conferred the ability to specifically bind to S. cerevisiae var. diastaticus cells, which flocculate, but not to Sigma1278b cells, which do not flocculate. This was true for the 196-kDa form as well as the high-molecular weight form of Flo11p, regardless of the strain source. The coated beads bound to S. cerevisiae var. diastaticus cells expressing FLO11 and failed to bind to cells with a deletion of FLO11, demonstrating a homotypic adhesive mechanism. Flo11p was shown to be a mannoprotein. Bead-to-cell adhesion was inhibited by mannose, which also inhibits Flo11-dependent flocculation in vivo, further suggesting that this in vitro system is a useful model for the study of fungal adhesion. PMID- 17921351 TI - Endocytosis in the shiitake mushroom Lentinula edodes and involvement of GTPase LeRAB7. AB - Endocytosis is the process by which substrates enter a cell without passing through the plasma membrane but rather invaginate the cell membrane and form intracellular vesicles. Rab7 regulates endocytic trafficking between early and late endosomes and between late endosomes and lysosomes. LeRab7 in Lentinula edodes is strongly homologous to Rab7 in Homo sapiens. Receptors for activated C kinase-1 (LeRACK1) and Rab5 GTPase (LeRAB5) were isolated as interacting partners of LeRab7, and the interactions were confirmed by in vivo and in vitro protein interaction assays. The three genes showed differential expression in the various developmental stages of the mushroom. In situ hybridization showed that the three transcripts were localized in regions of active growth, such as the outer region of trama cells, and the subhymenium of the hymenophore of mature fruiting bodies and the prehymenophore of young fruiting bodies. The existence of endocytosis in the mycelium and hymenophores was confirmed by the internalization of FM4-64. LeRAB7 was partially colocalized with the AM4-64 and was located in the late endocytic pathway. This is the first report of the presence of endocytosis in homobasidiomycetes. LeRAB7, LeRAB5, and LeRACK1 may contribute to the growth of L. edodes and cell differentiation in hymenophores. PMID- 17921352 TI - Trypanosoma brucei RNA binding proteins p34 and p37 mediate NOPP44/46 cellular localization via the exportin 1 nuclear export pathway. AB - We have previously identified and characterized two novel nuclear RNA binding proteins, p34 and p37, which have been shown to interact with a family of nucleolar phosphoproteins, NOPP44/46, in Trypanosoma brucei. These proteins are nearly identical, the major difference being an 18-amino-acid insert in the N terminus of p37. In order to characterize the interaction between p34 and p37 and NOPP44/46, we have utilized an RNA interference (RNAi) cell line that specifically targets p34 and p37. Within these RNAi cells, we detected a disruption of a higher-molecular-weight complex containing NOPP44/46, as well as a dramatic increase in nuclear NOPP44/46 protein levels. We demonstrated that no change occurred in NOPP44/46 mRNA steady-state levels or stability, nor was there a change in cellular protein levels. These results led us to investigate whether p34 and p37 regulate NOPP44/46 cellular localization. Examination of the p34 and p37 amino acid sequences revealed a leucine-rich nuclear export signal, which interacts with the nuclear export factor exportin 1. Immune capture experiments demonstrated that p34, p37, and NOPP44/46 associate with exportin 1. When these experiments were performed with p34/p37 RNAi cells, NOPP44/46 no longer associated with exportin 1. Sequential immune capture experiments demonstrated that p34, p37, NOPP44/46, and exportin 1 exist in a common complex. Inhibiting exportin 1-mediated nuclear export led to an increase in nuclear NOPP44/46 proteins, indicating that they are exported from the nucleus via this pathway. Together, our results demonstrate that p34 and p37 regulate NOPP44/46 cellular localization by facilitating their association with exportin 1. PMID- 17921353 TI - An Arabidopsis gene network based on the graphical Gaussian model. AB - We describe a gene network for the Arabidopsis thaliana transcriptome based on a modified graphical Gaussian model (GGM). Through partial correlation (pcor), GGM infers coregulation patterns between gene pairs conditional on the behavior of other genes. Regularized GGM calculated pcor between gene pairs among approximately 2000 input genes at a time. Regularized GGM coupled with iterative random samplings of genes was expanded into a network that covered the Arabidopsis genome (22,266 genes). This resulted in a network of 18,625 interactions (edges) among 6760 genes (nodes) with high confidence and connections representing approximately 0.01% of all possible edges. When queried for selected genes, locally coherent subnetworks mainly related to metabolic functions, and stress responses emerged. Examples of networks for biochemical pathways, cell wall metabolism, and cold responses are presented. GGM displayed known coregulation pathways as subnetworks and added novel components to known edges. Finally, the network reconciled individual subnetworks in a topology joined at the whole-genome level and provided a general framework that can instruct future studies on plant metabolism and stress responses. The network model is included. PMID- 17921354 TI - PennCNV: an integrated hidden Markov model designed for high-resolution copy number variation detection in whole-genome SNP genotyping data. AB - Comprehensive identification and cataloging of copy number variations (CNVs) is required to provide a complete view of human genetic variation. The resolution of CNV detection in previous experimental designs has been limited to tens or hundreds of kilobases. Here we present PennCNV, a hidden Markov model (HMM) based approach, for kilobase-resolution detection of CNVs from Illumina high-density SNP genotyping data. This algorithm incorporates multiple sources of information, including total signal intensity and allelic intensity ratio at each SNP marker, the distance between neighboring SNPs, the allele frequency of SNPs, and the pedigree information where available. We applied PennCNV to genotyping data generated for 112 HapMap individuals; on average, we detected approximately 27 CNVs for each individual with a median size of approximately 12 kb. Excluding common rearrangements in lymphoblastoid cell lines, the fraction of CNVs in offspring not detected in parents (CNV-NDPs) was 3.3%. Our results demonstrate the feasibility of whole-genome fine-mapping of CNVs via high-density SNP genotyping. PMID- 17921355 TI - Comparative analysis of chicken chromosome 28 provides new clues to the evolutionary fragility of gene-rich vertebrate regions. AB - The chicken genome draft sequence has provided a valuable resource for studies of an important agricultural and experimental model species and an important data set for comparative analysis. However, some of the most gene-rich segments are missing from chicken genome draft assemblies, limiting the analysis of a substantial number of genes and preventing a closer look at regions that are especially prone to syntenic rearrangements. To facilitate the functional and evolutionary analysis of one especially gene-rich, rearrangement-prone genomic region, we analyzed sequence from BAC clones spanning chicken microchromosome GGA28; as a complement we also analyzed a gene-sparse, stable region from GGA11. In these two regions we documented the conservation and lineage-specific gain and loss of protein-coding genes and precisely mapped the locations of 31 major human chicken syntenic breakpoints. Altogether, we identified 72 lineage-specific genes, many of which are found at or near syntenic breaks, implicating evolutionary breakpoint regions as major sites of genetic innovation and change. Twenty-two of the 31 breakpoint regions have been reused repeatedly as rearrangement breakpoints in vertebrate evolution. Compared with stable GC matched regions, GGA28 is highly enriched in CpG islands, as are break-prone intervals identified elsewhere in the chicken genome; evolutionary breakpoints are further enriched in GC content and CpG islands, highlighting a potential role for these features in genome instability. These data support the hypothesis that chromosome rearrangements have not occurred randomly over the course of vertebrate evolution but are focused preferentially within "fragile" regions with unusual DNA sequence characteristics. PMID- 17921356 TI - Determinants of whole-body protein metabolism in subjects with and without type 2 diabetes. AB - OBJECTIVE: Whole-body protein metabolism is abnormal in suboptimally controlled type 2 diabetes and obesity. We hypothesized that glycemia, insulin resistance, and waist circumference modulate these alterations in type 2 diabetes and, to a lesser extent, in individuals without type 2 diabetes. RESEARCH DESIGN AND METHODS: In 88 lean and obese subjects without and 40 with type 2 diabetes on an inpatient protein-controlled isoenergetic diet for 7 days, whole-body protein turnover was measured using the fed-fasted 60-h oral (15)N-glycine method. Nitrogen flux was determined from urinary (15)N urea and protein synthesis, breakdown and net balance calculated. Indexes of diabetes control, resting energy expenditure (REE), and body composition were assessed. RESULTS: Higher protein turnover in obese subjects was further increased, and net balance was lower in type 2 diabetes. Waist-to-hip ratio and ln homeostasis model assessment of insulin resistance (HOMA-IR) explained 40% of the variance in flux in type 2 diabetes; fat-free mass and lnHOMA-IR explained 62% in subjects without type 2 diabetes. Overall, fasting glucose explained 16% of the variance in net balance. In type 2 diabetes, net balance correlated negatively with fasting glucose in men and positively with hip circumference in women. CONCLUSIONS: Kinetics of whole body protein metabolism are elevated, and net balance is diminished in type 2 diabetes, independently of obesity. Elevated flux is associated with greater visceral adiposity, REE, and insulin resistance of glucose. In type 2 diabetic men, these alterations worsened with magnitude of hyperglycemia. In type 2 diabetic women, larger hip circumferences may protect against such alterations. Our findings suggest that dietary protein requirements may be greater in type 2 diabetes to offset a reduced net balance, aggravated as glycemia increases, especially in men. PMID- 17921357 TI - Prevalence of diabetes and impaired fasting glucose in the adult population of Iran: National Survey of Risk Factors for Non-Communicable Diseases of Iran. AB - OBJECTIVE: Despite concerns regarding a diabetes epidemic in the Middle East, internationally published data on national estimates of prevalent type 2 diabetes in Iran do not exist. With this article, we document a dramatically high prevalence of diabetes in Iran. RESEARCH DESIGN AND METHODS: Our data are based on the results of the first Survey of Risk Factors of Non-Communicable Diseases of Iran, 2005. In this national cross-sectional survey, 70,981 Iranian citizens aged 25-64 years were recruited. RESULTS: We found that 7.7% of adults aged 25-64 years, or 2 million adults, have diabetes, among whom one-half are undiagnosed. An additional 16.8%, or 4.4 million, of Iranian adults have impaired fasting glucose. CONCLUSIONS: The high prevalence of diabetes in working-age adults is an ominous sign for this developing nation. As the relatively young Iranian population ages in the future and urbanization continues or accelerates, the prevalence of diabetes will likely escalate. PMID- 17921358 TI - Conditional deletion of Pten causes bronchiolar hyperplasia. AB - Tumor suppressor phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a lipid phosphatase that regulates multiple cellular processes including cell polarity, migration, proliferation, and carcinogenesis. In this work, we demonstrate that conditional deletion of Pten (Pten(Delta/Delta)) in the respiratory epithelial cells of the developing mouse lung caused epithelial cell proliferation and hyperplasia as early as 4 to 6 weeks of age. While bronchiolar cell differentiation was normal, as indicated by beta-tubulin and FOXJ1 expression in ciliated cells and by CCSP expression in nonciliated cells, cell proliferation (detected by expression of Ki-67, phospho-histone-H3, and cyclin D1) was increased and associated with activation of the AKT/mTOR survival pathway. Deletion of Pten caused papillary epithelial hyperplasia characterized by a hypercellular epithelium lining papillae with fibrovascular cores that protruded into the airway lumens. Cell polarity, as assessed by subcellular localization of cadherin, beta-catenin, and zonula occludens-1, was unaltered. PTEN is required for regulation of epithelial cell proliferation in the lung and for the maintenance of the normal simple columnar epithelium characteristics of bronchi and bronchioles. PMID- 17921359 TI - A functional and regulatory map of asthma. AB - The prevalence and morbidity of asthma, a chronic inflammatory airway disease, is increasing. Animal models provide a meaningful but limited view of the mechanisms of asthma in humans. A systems-level view of asthma that integrates multiple levels of molecular and functional information is needed. For this, we compiled a gene expression compendium from five publicly available mouse microarray datasets and a gene knowledge base of 4,305 gene annotation sets. Using this collection we generated a high-level map of the functional themes that characterize animal models of asthma, dominated by innate and adaptive immune response. We used Module Networks analysis to identify co-regulated gene modules. The resulting modules reflect four distinct responses to treatment, including early response, general induction, repression, and IL-13-dependent response. One module with a persistent induction in response to treatment is mainly composed of genes with suggested roles in asthma, suggesting a similar role for other module members. Analysis of IL-13-dependent response using protein interaction networks highlights a role for TGF-beta1 as a key regulator of asthma. Our analysis demonstrates the discovery potential of systems-level approaches and provides a framework for applying such approaches to asthma. PMID- 17921360 TI - Cyclic stretch of human lung cells induces an acidification and promotes bacterial growth. AB - The reasons for bacterial proliferation in the lungs of mechanically ventilated patients are poorly understood. We hypothesized that prolonged cyclic stretch of lung cells influenced bacterial growth. Human alveolar type II-like A549 cells were submitted in vitro to prolonged cyclic stretch. Bacteria were cultured in conditioned supernatants from cells submitted to stretch and from control static cells. Escherichia coli had a marked growth advantage in conditioned supernatants from stretched A549 cells, but also from stretched human bronchial BEAS-2B cells, human MRC-5 fibroblasts, and murine RAW 264.7 macrophages. Stretched cells compared with control static cells acidified the milieu by producing increased amounts of lactic acid. Alkalinization of supernatants from stretched cells blocked E. coli growth. In contrast, acidification of supernatants from control cells stimulated bacterial growth. The effect of various pharmacological inhibitors of metabolic pathways was tested in this system. Treatment of A549 cells and murine RAW 264.7 macrophages with the Na(+)/K(+)-ATPase pump inhibitor ouabain during cyclic stretch blocked both the acidification of the milieu and bacterial growth. Several pathogenic bacteria originating from lungs of patients with ventilator-associated pneumonia (VAP) also grow better in vitro at slightly acidic pH (pH 6-7.2), pH similar to those measured in the airways from ventilated patients. This novel metabolic pathway stimulated by cyclic stretch may represent an important pathogenic mechanism of VAP. Alkalinization of the airways may represent a promising preventive strategy in ventilated critically ill patients. PMID- 17921361 TI - How bad is fructose? PMID- 17921362 TI - Folic acid fortification: is masking of vitamin B-12 deficiency what we should really worry about? PMID- 17921363 TI - Potential role of sugar (fructose) in the epidemic of hypertension, obesity and the metabolic syndrome, diabetes, kidney disease, and cardiovascular disease. AB - Currently, we are experiencing an epidemic of cardiorenal disease characterized by increasing rates of obesity, hypertension, the metabolic syndrome, type 2 diabetes, and kidney disease. Whereas excessive caloric intake and physical inactivity are likely important factors driving the obesity epidemic, it is important to consider additional mechanisms. We revisit an old hypothesis that sugar, particularly excessive fructose intake, has a critical role in the epidemic of cardiorenal disease. We also present evidence that the unique ability of fructose to induce an increase in uric acid may be a major mechanism by which fructose can cause cardiorenal disease. Finally, we suggest that high intakes of fructose in African Americans may explain their greater predisposition to develop cardiorenal disease, and we provide a list of testable predictions to evaluate this hypothesis. PMID- 17921364 TI - Birth weight; postnatal, infant, and childhood growth; and obesity in young adulthood: evidence from the Barry Caerphilly Growth Study. AB - BACKGROUND: Birth weight has been shown to be positively associated with adult obesity, but relatively few studies have examined the associations with growth in specific periods of early childhood. OBJECTIVE: The objective was to assess the association of measures of growth between birth and 5 y of age with adult measures of adiposity. DESIGN: We conducted a longitudinal study of young adults from Barry and Caerphilly, United Kingdom, who had previously taken part between 1972 and 1974 in a randomized controlled trial of milk supplementation. We reexamined 679 men and women (72% of the target population) to measure body mass index (BMI; in kg/m(2)), waist-to-hip ratio, sagittal abdominal diameter, and waist circumference. RESULTS: An increase in weight velocity from 1 y and 9 mo to 5 y of age was the most important predictor of BMI, waist circumference, and sagittal abdominal diameter. A z-score increase in weight gain in this period was associated with an increase in BMI of 1.13 (95% CI: 0.69, 1.57; P < 0.001). Infant weight gain from 5 mo to 1 y and 9 mo was the strongest predictor of waist to-hip ratio (0.51; 95% CI: 0.00, 1.02; P = 0.05). CONCLUSIONS: Birth weight does not predict adiposity on the basis of weight gain in childhood. The association between adult adiposity and weight gain in different periods is variable and depends on the measure of adiposity that is used. It remains unclear whether early childhood is the optimum period in the life course for the primary prevention of adult adiposity. PMID- 17921366 TI - Effects of dietary composition on postprandial endothelial function and adiponectin concentrations in healthy humans: a crossover controlled study. AB - BACKGROUND: Abnormalities during the postprandial state contribute to the development of atherosclerosis. Reportedly, postprandial hyperglycemia, hypertriglyceridemia, and hyperlipacidemia independently cause postprandial cytokine activation. However, it is not clear which dietary composition preferentially affects postprandial endothelial function in healthy subjects. OBJECTIVE: We aimed to examine the associations of dietary composition and postprandial endothelial function in healthy subjects. DESIGN: The effects of a single ingestion of a high-carbohydrate meal (300 kcal, 100% carbohydrate), a high-fat meal (30 g fat/m(2), 35% fat), or a standard test meal (478 kcal; 16.4% protein, 32.7% fat, 50.4% carbohydrate) on postprandial plasma concentrations of adiponectin and forearm blood flow (FBF) during reactive hyperemia were studied in healthy subjects. RESULTS: The peak FBF response and the total reactive hyperemic flow (flow debt repayment; FDR), indexes of resistance artery endothelial function, were unchanged after ingestion of a high-carbohydrate and standard test meal but decreased 120 and 240 min after a high-fat meal. After a high-fat meal, decreases in peak FBF and FDR were well correlated with an increase in plasma free fatty acid (FFA) concentrations but not with the other biochemical variables, including triacylglycerol, insulin, glucose, total cholesterol, HDL cholesterol, and adiponectin. CONCLUSIONS: Postprandial endothelial function was impaired only after the high-fat diet and not after the high-carbohydrate or standard test meal in healthy subjects. Because such endothelial dysfunction after a high-fat meal was closely correlated with FFA concentrations, postprandial state could be hazardous, mostly through acute hyperlipacidemia in healthy subjects. PMID- 17921365 TI - Price and maternal obesity influence purchasing of low- and high-energy-dense foods. AB - BACKGROUND: Price can influence food purchases, which can influence consumption. Limited laboratory research has assessed the effect of price changes on food purchases, and no research on individual differences that may interact with price to influence purchases exists. OBJECTIVE: We aimed to assess the influence of price changes of low-energy-density (LED) and high-energy-density (HED) foods on mother's food purchases in a laboratory food-purchasing analogue. DESIGN: Mothers were randomly assigned to price conditions in which the price of either LED or HED foods was manipulated from 75% to 125% of the reference purchase price, whereas the price of the alternative foods was kept at the reference value. Mothers completed purchases for 2 income levels ($15 or $30 per family member). RESULTS: Purchases were reduced when prices of LED (P < 0.01) and HED (P < 0.001) foods were increased. Maternal BMI interacted with price to influence purchases of HED foods when the price of HED foods increased (P = 0.016) and interacted with price to influence purchases of LED foods when the price of HED foods increased (P = 0.008). CONCLUSION: These results show the relevance of considering price change as a way to influence food purchases of LED compared with HED foods and the possibility that individual differences may influence the own-price elasticity of HED foods and substitution of LED for HED foods. PMID- 17921367 TI - Plasma and erythrocyte biomarkers of dairy fat intake and risk of ischemic heart disease. AB - BACKGROUND: The relation between dairy product intake and the risk of ischemic heart disease (IHD) remains controversial. OBJECTIVE: We aimed to explore biomarkers of dairy fat intake in plasma and erythrocytes and to assess the hypothesis that higher concentrations of these biomarkers are associated with a greater risk of IHD in US women. DESIGN: Among 32,826 participants in the Nurses' Health Study who provided blood samples in 1989-1990, 166 incident cases of IHD were ascertained between baseline and 1996. These cases were matched with 327 controls for age, smoking, fasting status, and date of blood drawing. RESULTS: Among controls, correlation coefficients between average dairy fat intake in 1986 1990 and 15:0 and trans 16:1n-7 content were 0.36 and 0.30 for plasma and 0.30 and 0.32 for erythrocytes, respectively. In multivariate analyses, with control for age, smoking, and other risk factors of IHD, women with higher plasma concentrations of 15:0 had a significantly higher risk of IHD. The multivariate adjusted relative risks (95% CI) from the lowest to highest tertile of 15:0 concentrations in plasma were 1.0 (reference), 2.18 (1.20, 3.98), and 2.36 (1.16, 4.78) (P for trend = 0.03). Associations for other biomarkers were not significant. CONCLUSIONS: Plasma and erythrocyte contents of 15:0 and trans 16:1n 7 can be used as biomarkers of dairy fat intake. These data suggest that a high intake of dairy fat is associated with a greater risk of IHD. PMID- 17921368 TI - Isoflavone intake in persons at high risk of cardiovascular events: implications for vascular endothelial function and the carotid atherosclerotic burden. AB - BACKGROUND: Epidemiologic studies have suggested that a high phytoestrogen intake is inversely associated with cardiovascular disease risk factors and the incidence of cardiovascular events. However, the relation between the intake of isoflavone, a major component of phytoestrogen, and vascular endothelial function and the atherosclerotic burden remains unclear. OBJECTIVE: We aimed to investigate the effects of various dietary soy isoflavone intakes on brachial artery flow-mediated dilation and mean maximum carotid intima-media thickness. DESIGN: We studied 126 consecutive patients (x +/- SD age: 66.5 +/- 11.1 y; 69% male) at high risk of cardiovascular events (94% had documented coronary artery disease or stroke; 44% had diabetes). A validated food-frequency questionnaire was used to estimate their dietary intake after they achieved stable dietary patterns for 3 mo. RESULTS: The median intakes of isoflavone and soy protein were 5.5 (range: 2.2-13.3) mg/d and 1.2 (range: 0.4-2.8) g/d, respectively. Persons in the 4th quartile of daily isoflavone intake had significantly (P < 0.05) greater flow-mediated dilation-but not mean maximum carotid intima-media thickness-than did those in the lower quartiles. After adjustment for potential confounders, a higher isoflavone intake in the 4th than in the 1st quartile (>13.3 mg/d), but not a higher intake of soy protein, predicted an absolute 2.71% increase in flow mediated dilation (relative increase: 103%; P = 0.02) and a 0.17-mm decrease in mean maximum carotid intima-media thickness (relative decrease: 14.5%; P = 0.04). CONCLUSION: In persons at high risk of cardiovascular events, a greater isoflavone intake is associated with better vascular endothelial function and lower carotid atherosclerotic burden. PMID- 17921369 TI - Effects of moderate variations in the macronutrient content of the diet on cardiovascular disease risk factors in obese patients with the metabolic syndrome. AB - BACKGROUND: The metabolic syndrome is a cluster of abnormalities that is accompanied by a 2-fold increase in the risk of cardiovascular disease. Even if there is full agreement that lifestyle changes to induce weight loss are the first-line approach, the ideal diet for the treatment of the metabolic syndrome remains uncertain. OBJECTIVE: The objective was to compare the effects of 2 diets on cardiovascular disease risk factors in obese patients with the metabolic syndrome. DESIGN: The study was carried out in 100 patients randomly assigned to either a diet relatively rich in carbohydrate [65% of energy as carbohydrate, 13% as protein, and 22% as fat (17% as unsaturated fat)] or a diet that was low in carbohydrate and high in protein and in monounsaturated fat [48% of energy as carbohydrate, 19% as protein, and 33% as fat (24% as unsaturated fat)]. RESULTS: All 100 patients completed the 5-mo study. At the end of the study, all the components of the metabolic syndrome (except HDL, which did not change) decreased significantly in both groups. With the high-carbohydrate diet, a significant decrease in LDL-cholesterol concentrations was also observed. Although the extent of the resolution of the metabolic syndrome was not different between groups, the low-carbohydrate diet was associated with a greater decrease in the prevalence of hypertension (P < 0.05) and of hypertriacylglycerolemia (P < 0.001). CONCLUSION: Tailoring diet interventions to the specific presentation of the metabolic syndrome may be the best way of reducing the risk factors for cardiovascular disease. PMID- 17921370 TI - Estimation of thigh muscle cross-sectional area by dual-energy X-ray absorptiometry in frail elderly patients. AB - BACKGROUND: Thigh muscle mass and cross-sectional area (CSA) are useful indexes of sarcopenia and the response to treatment in older patients. Current criterion methods are computed tomography (CT) and magnetic resonance imaging. OBJECTIVE: The objective was to compare thigh muscle mass estimated by dual-energy X-ray absorptiometry (DXA), a less expensive and more accessible method, with thigh muscle CSA determined by CT in a group of elderly patients recovering from hip fracture. DESIGN: Midthigh muscle CSA (in cm(2)) was assessed from a 1-mm CT slice and midthigh muscle mass (g) from a 1.3-cm DXA slice in 30 patients (24 women) aged 81 +/- 8 y during 12 mo of follow-up. Fat-to-lean soft tissue ratios were calculated with each technique to permit direct comparison of a variable in the same units. RESULTS: Baseline midthigh muscle CSA was highly correlated with midthigh muscle mass (r = 0.86, P < 0.001) such that DXA predicted CT-determined CSA with an SEE of 10 cm(2) (an error of approximately 12% of the mean CSA value). CT- and DXA-determined ratios of midthigh fat to lean mass were similarly related (intraclass correlation coefficient = 0.87, P < 0.001). When data were expressed as the changes from baseline to follow-up, CT and DXA changes were weakly correlated (intraclass correlation coefficient = 0.51, P = 0.019). CONCLUSIONS: Assessment of sarcopenia by DXA midthigh slice is a potential low radiation, accessible alternative to CT scanning of older patients. The errors inherent in this technique indicate, however, that it should be applied to groups of patients rather than to individuals or to evaluate the response to interventions. PMID- 17921371 TI - The effects of seasonal variation of 25-hydroxyvitamin D and fat mass on a diagnosis of vitamin D sufficiency. AB - BACKGROUND: The effect of season on vitamin D status is often overlooked in studies of optimal vitamin D concentrations and in clinical practice. OBJECTIVES: We aimed to determine the effects of seasonal variation of 25-hydroxyvitamin D [25(OH)D] on a previously selected minimum concentration for vitamin D sufficiency (50 nmol/L) and to determine whether fat mass modifies these effects. DESIGN: A cross-sectional study evaluated 1606 healthy postmenopausal women and 378 older men from Auckland, New Zealand, who were undergoing single measurements of 25(OH)D. RESULTS: Concentrations of <50 nmol 25(OH)D/L were seen in 49% (range: 23%-74%) of women and 9% (range: 0%-26%) of men when measured, but 73% of women and 39% of men were predicted to have trough 25(OH)D concentrations < 50 nmol/L, according to the demonstrated seasonal variation. The predicted duration of 25(OH)D concentrations < 50 nmol/L was 250 d/y in women and 165 d/y in men. CONCLUSION: Seasonal variation significantly affects the diagnosis of vitamin D sufficiency, which requires seasonally adjusted thresholds individualized for different locations. Clinicians should consider the month of sampling and the amount of body fat when interpreting 25(OH)D measurements. PMID- 17921372 TI - Effects of overfeeding on the neuronal response to visual food cues. AB - BACKGROUND: The regulation of energy intake is a complex process involving the integration of homeostatic signals and both internal and external sensory inputs. Environmental visual cues are one of the first and primary inputs signaling the potential availability of food. OBJECTIVE: We examined the effects of short-term overfeeding on the neuronal responses to food-related visual stimuli in thin individuals. DESIGN: Twenty-five thin individuals (13 women, 12 men) were studied. Functional magnetic resonance imaging (fMRI) was performed after 2 days of eucaloric energy intake and after 2 days of 30% overfeeding in a counterbalanced design. fMRI was performed while the subjects were presented with visual stimuli in 3 different categories: neutral control objects, foods of neutral hedonic value, and foods of high hedonic value. Measures of appetite were obtained by using visual analogue scales before and after meals. RESULTS: In the eucaloric state, pictures of foods of high hedonic value elicited greater activation of neuronal regions than did neutrally rated foods, which is consistent with visual processing and attention (inferior temporal visual cortex, posterior parietal cortex, premotor cortex, and hippocampus) and with activation of the hypothalamus. Two days of overfeeding led to significant attenuation of these responses. Overfeeding also resulted in reduced hunger ratings and increased satiety ratings. CONCLUSION: These findings emphasize the important role of external food-related visual cues and suggest that there are interactions between external visual sensory inputs, energy balance status, and brain regions important in the homeostatic regulation of energy intake. PMID- 17921373 TI - Insoluble cereal fiber reduces appetite and short-term food intake and glycemic response to food consumed 75 min later by healthy men. AB - BACKGROUND: Insoluble fiber consumption is associated with reduced risk of obesity and diabetes, but its mechanisms of action are unknown. OBJECTIVE: The objective was to describe the effect of insoluble fiber on appetite, short-term food intake, and blood glucose (BG) before and after a meal 75 min later in healthy men. DESIGN: In a crossover design, high-fiber (HF; 33 g insoluble fiber) cereal, low-fiber (LF) cereal, white bread (WB), and water control were administered to young men after an overnight fast. Caloric treatments had similar energy, macronutrient content, volume, and weight. In the first experiment, subjective appetite and BG were measured at 15-min intervals before and after an ad libitum meal at 75 min. In the second experiment, a preset pizza meal (850 kcal) was consumed at 75 min. Appetite and blood glucose were measured for 150 min at fasting and at 15-min intervals before and after the fixed meal. RESULTS: In experiment 1, ad libitum food intake was lower after the HF cereal and WB than after the LF cereal and water (937 +/- 86, 970 +/- 65, 1109 +/- 90, 1224 +/- 89 kcal, respectively; P < 0.001). Appetite was lower (P < 0.05) after the HF cereal than after the WB but not different from the LF cereal. The BG area under the curve (AUC) did not differ among the HF cereal, WB, and LF cereal from 0 to 75 min, but the postmeal BG increased after the WB and LF cereal but not after the HF cereal. In experiment 2, the HF cereal, but not the LF cereal or WB, increased fullness before and prevented an increase in the BG AUC after the preset meal (P < 0.05). CONCLUSION: A serving of 33 g insoluble fiber reduced appetite, lowered food intake, and reduced glycemic response to a meal consumed 75 min later. PMID- 17921374 TI - Breakfast glycemic index affects subsequent daily energy intake in free-living healthy children. AB - BACKGROUND: Experimental studies have reported that the effect of a meal's glycemic index (GI) on subsequent energy intake depends on the timing of the subsequent meal. OBJECTIVE: We examined whether the timing of the next meal after breakfast modifies the effect of the breakfast GI (GI(br)) on subsequent daytime energy intake of healthy free-living children. DESIGN: Analyses included 381 participants of the DOrtmund Nutritional and Anthropometrical Longitudinally Designed (DONALD) Study who had provided weighed dietary records at ages 2, 4-5, and 7 y. RESULTS: At all ages, among children who consumed their next meal in the early postprandial phase (after 3-4 h), children with a lower GI(br) consumed more calories throughout the remainder of the day than did children with a higher GI(br), independent of major dietary confounders. For the age groups 2, 4-5, and 7 y, energy intakes in tertiles 1 and 3 were 785 kcal (95% CI: 743-830 kcal) and 717 kcal (678-758 kcal), P for trend = 0.2; 993 kcal (941-1047 kcal) and 949 kcal (900-1000 kcal), P for trend = 0.05; 1255 (1171-1344) and 1166 (1090-1247 kcal), P for trend = 0.03, respectively. Conversely, among children consuming their next meal in the late postprandial phase (>3-4 h), subsequent daytime energy intake was not associated with GI(br). CONCLUSION: This study confirms differential early and late postprandial effects of the GI(br) on subsequent daytime energy intake for free-living children at different ages. Interestingly, the apparent short-term satiating effect of a higher GI(br), in particular, persisted throughout the day, if a second breakfast was consumed midmorning. PMID- 17921375 TI - Dietary glycemic index and glycemic load are associated with high-density lipoprotein cholesterol at baseline but not with increased risk of diabetes in the Whitehall II study. AB - BACKGROUND: Findings of the effect of dietary glycemic index (GI) and glycemic load (GL) on the risk of incident diabetes are inconsistent. OBJECTIVE: We examined the associations of dietary GI and GL with clinical variables at baseline and the incidence of diabetes. DESIGN: The 7321 white Whitehall II participants (71% men) attending screening in 1991-1993, free of diabetes at baseline, and with food-frequency questionnaire data were followed for 13 y. RESULTS: At baseline, dietary GI and GL were associated inversely with HDL cholesterol, and GI was associated directly with triacylglycerols. Dietary GI and GL were related inversely to fasting glucose and directly to 2-h postload glucose, but only the association between GI and 2-h postload glucose was robust to statistical adjustments for employment grade, physical activity, smoking status, and intakes of alcohol, fiber, and carbohydrates. High-dietary GI was not associated with increased risk of incident diabetes. Hazard ratios (HRs) across sex-specific tertiles of dietary GI were 1.00, 0.95 (95% CI: 0.73, 1.24), and 0.94 (95% CI: 0.72, 1.22) (adjusted for sex, age, and energy misreporting; P for trend = 0.64). Corresponding HRs across tertiles of dietary GL were 1.00, 0.92 (95% CI: 0.71, 1.19), and 0.70 (95% CI: 0.54, 0.92) (P for trend = 0.01). The protective effect on diabetes risk remained significant after adjustment for employment grade, smoking, and alcohol intake but not after further adjustment for carbohydrate and fiber intakes. CONCLUSION: The proposed protective effect of low-dietary GI and GL diets on diabetes risk could not be confirmed in this study. PMID- 17921376 TI - Reevaluation of the protein requirement in young men with the indicator amino acid oxidation technique. AB - BACKGROUND: The current estimated protein requirements are based on the nitrogen balance method, which has many limitations. An alternate approach is needed to permit a reevaluation of protein requirements. OBJECTIVE: The objective was to determine protein requirements in men by using the indicator amino acid oxidation technique. DESIGN: Eight healthy men randomly received graded protein intakes (0.10, 0.30, 0.60, 0.90, 1.2, 1.5 and 1.8 g kg(-1) d(-1)) as a crystalline amino acid mixture along with L-[1-(13)C]phenylalanine. The mean protein requirement was determined by applying a biphase linear regression crossover analysis on F(13)CO(2) data, which identified a breakpoint at the minimal rate of appearance of (13)CO(2) to graded protein intakes. RESULTS: The mean and population-safe (recommended dietary allowance; RDA) protein requirements were found to be 0.93 and 1.2 g kg(-1) d(-1), respectively. These requirements are comparable with those estimated by the application of a biphase linear regression model to the data from nitrogen balance studies (0.91 and 1.0 g kg(-1) d(-1), respectively). These requirements are 41% and 50% higher than the current recommendations for the estimated average requirement (EAR) of 0.66 g kg(-1) d(-1) and the RDA of 0.80 g kg(-1) d(-1), as determined by applying a linear regression model where it intersects the zero balance line. CONCLUSION: The indicator amino acid oxidation technique defined a protein requirement that is comparable with that estimated by the application of a biphase linear regression model to nitrogen balance data in the literature. Our data and the reanalysis of the preexisting nitrogen balance data suggest that the current recommended protein requirements are too low and require reassessment. PMID- 17921377 TI - Albumin synthesis in premature neonates is stimulated by parenterally administered amino acids during the first days of life. AB - BACKGROUND: We recently showed that parenteral administration of amino acids to premature infants immediately after birth is safe and results in a positive nitrogen balance and increased whole-body protein synthesis. However, we did not determine organ-specific effects; albumin, produced by the liver, is an important protein, but its concentration is often low in premature neonates during the first few days after birth. OBJECTIVE: The objective of the study was to test the hypothesis that the fractional and absolute albumin synthesis rates would increase with the administration of amino acids after birth, even at low nonprotein energy intake. DESIGN: Premature infants (<1500 g birth weight), who were on ventilation, received from birth onward either glucose only (control group, n = 7) or glucose and 2.4 g amino acid kg(-1) d(-1) (intervention group, n = 8). On postnatal day 2, all infants received a primed continuous infusion of [1 (13)C]leucine, and mass spectrometry techniques were used to determine the incorporation of the leucine into albumin. Results are expressed as medians and 25th and 75th percentiles. RESULTS: Albumin fractional synthesis rates in the intervention group were significantly higher than those in the control group [22.9% (17.6-28.0%)/d and 12.6% (11.0-19.4%)/d, respectively; P = 0.029]. Likewise, the albumin absolute synthesis rates in the intervention group were significantly higher than those in the control group [228 (187-289) mg kg(-1) d( 1) and 168 (118-203) mg kg(-1) d(-1), respectively; P = 0.030]. CONCLUSION: Amino acid administration increases albumin synthesis rates in premature neonates even at a low energy intake. PMID- 17921378 TI - Short-term overfeeding increases resting energy expenditure in patients with HIV lipodystrophy. AB - BACKGROUND: HIV lipodystrophy and other lipodystrophy syndromes are characterized by extensive loss of subcutaneous adipose tissue. Lipodystrophy syndromes are also associated with increased resting energy expenditure (REE). This hypermetabolism may be an adaptive response to an inability to store triacylglycerol fuel in a normal manner. OBJECTIVE: This study was done to determine whether REE increases significantly after short-term overfeeding in patients with HIV lipodystrophy. DESIGN: REE was measured in HIV-infected patients with lipodystrophy (n = 9) and in HIV-infected (n = 10) and healthy (n = 9) controls after 3 d on a eucaloric diet and again after 3 d on a diet of similar composition but increased in calories by 50%. RESULTS: After 3 d of eucaloric feeding, REE was significantly higher in patients with HIV lipodystrophy [33.2 +/- 0.27 kcal/kg lean body mass (LBM)] than for both HIV infected and healthy controls (29.9 +/- 0.26 and 29.6 +/- 0.27 kcal/kg LBM, respectively; P < 0.01). Furthermore, after 3 d of overfeeding, REE increased significantly in patients with HIV lipodystrophy but not in the control groups (33.2 +/- 0.27 vs 34.7 +/- 0.27 kcal/kg LBM; P < 0.01). Finally, postprandial thermogenesis did not differ among the groups after a "normal" test meal but tended to be higher in patients with HIV lipodystrophy than in healthy controls after a large test meal. CONCLUSIONS: Adaptive thermogenesis in the resting component of total daily energy expenditure and in the postprandial period may be a feature of the HIV lipodystrophy syndrome and may be due to an inability to store triacylglycerol fuel in a normal manner. PMID- 17921379 TI - Diurnal variation in glutathione and cysteine redox states in human plasma. AB - BACKGROUND: Plasma glutathione/glutathione disulfide (GSH/GSSG) and cysteine/cystine (Cys/CySS) couples are oxidized in humans in association with oxidative stress and cardiovascular disease risk. Animal studies show that both pools undergo diurnal variations associated with dietary intake of sulfur amino acids. OBJECTIVE: The objective of this study was to determine whether the redox state of GSH, Cys, GSH/GSSG, or Cys/CySS undergoes diurnal variation in healthy adults. DESIGN: Plasma samples were collected every hour for 24 h from 63 persons aged 18-86 y who were consuming normal food (protein, 0.8 g kg(-1) d(-1); sulfur amino acids, 20 mg kg(-1) d(-1)) at standardized mealtimes. Measurements of Cys, CySS, GSH, and GSSG were used with the Nernst equation to calculate the redox states. RESULTS: Plasma Cys and GSH concentrations varied with the time of day. The highest values for plasma Cys occurred approximately 3 h after meals. Glutathione was maximal 6 h after peak plasma Cys. The calculated redox states of the GSH/GSSG and Cys/CySS couples varied in association with the concentrations of the thiol forms. Maximal reduction and oxidation of the Cys/CySS couple occurred at 2130 and 0630, whereas the respective values for the GSH/GSSG couple occurred at 0330 and 1330. The mean diurnal variation for Cys/CySS redox in persons aged >or=60 y was 1.8-fold that in persons aged <40 y. CONCLUSIONS: Cys/CySS and GSH/GSSG redox states in human plasma undergo diurnal variation with an increased magnitude of variation in Cys/CySS redox state in older persons. This variation could alter sensitivity to oxidative stress over a course of hours. PMID- 17921380 TI - Arginine flux and intravascular nitric oxide synthesis in severe childhood undernutrition. AB - BACKGROUND: Although nutritionally dispensable amino acids are not essential in the diet, adequate synthesis is necessary for maintenance of good health. Whereas children with edematous severe childhood undernutrition (SCU) can maintain production rates of glycine and serine despite a slower body protein breakdown rate, it is unknown whether the same is true for the semidispensable amino acid arginine. OBJECTIVE: We aimed to measure arginine flux and intravascular nitric oxide synthesis in children with SCU. DESIGN: Arginine flux and the fractional and absolute synthesis rates of plasma nitrite plus nitrate were measured postabsorptively by using a 6-h infusion of [(15)N(2)]-arginine in 2 groups of children with edematous (n = 14) or nonedematous (n = 7) SCU when they were infected and malnourished (postadmission day approximately 3; clinical phase 1), when they were no longer infected (postadmission day approximately 15; clinical phase 2), and when they were recovered (postadmission day approximately 55; clinical phase 3). RESULTS: Arginine flux was slower (P < 0.01) and plasma arginine concentrations were lower in the edematous group than in the nonedematous group at clinical phase 1. At clinical phase 2, flux doubled to a value that was not significantly different from the value at clinical phase 3. There were no significant differences in the plasma concentration or fractional or absolute synthesis rate of plasma nitrite plus nitrate between the groups at any clinical phase and among clinical phases within each group. CONCLUSION: Whereas children with nonedematous SCU can maintain arginine flux at the same rate as when recovered, children with edematous SCU cannot. The slower arginine flux was not, however, associated with slower nitric oxide synthesis. PMID- 17921381 TI - Effect of vitamin A supplementation with BCG vaccine at birth on vitamin A status at 6 wk and 4 mo of age. AB - BACKGROUND: The effect of vitamin A supplementation (VAS) at birth on subsequent vitamin A status has not been studied. OBJECTIVE: The objective was to study the effect of 50,000 IU vitamin A administered with BCG vaccine at birth on vitamin A status in both sexes. DESIGN: Within a randomized placebo-controlled trial of VAS, we obtained blood from 614 children at 6 wk of age and from 369 mother infant pairs at 4 mo of age. We assessed vitamin A status on the basis of serum retinol-binding protein (RBP) and measured serum C-reactive protein to monitor for concurrent infections. RESULTS: RBP concentrations indicated vitamin A deficiency in 32% of the children at age 6 wk and in 16% at age 4 mo. VAS was not associated with higher RBP concentrations overall or in either sex. However, the effect of VAS varied with maternal education (P for interaction = 0.004): At age 6 wk, VAS was associated with higher (9%; 95% CI: 2, 17%) RBP concentrations in children of noneducated mothers but not in children of educated mothers. Overall, RBP concentrations increased between 6 wk and 4 mo of age. The increase correlated inversely with the number of diphtheria-tetanus-pertussis (DTP) vaccines received in the interval (P = 0.009), particularly in girls (P for interaction = 0.01) and in vitamin A recipients (P = 0.01). CONCLUSIONS: Overall, VAS at birth had no effect on vitamin A status. However VAS may temporarily improve vitamin A status in the subgroup of children of noneducated mothers. In vitamin A recipients, subsequent DTP vaccines affected vitamin A status negatively. The main trial was registered at clinicaltrials.gov as NCT00168597. PMID- 17921382 TI - Vitamin A supplementation in iodine-deficient African children decreases thyrotropin stimulation of the thyroid and reduces the goiter rate. AB - BACKGROUND: Vitamin A (VA) deficiency (VAD) and iodine deficiency (ID) often coexist in children in Africa. VAD may affect thyroid function and the response to iodine prophylaxis. OBJECTIVE: The aim was to investigate the effects of supplementation with iodine or VA alone, and in combination, in children with concurrent VAD and ID. DESIGN: A 6-mo randomized, double-blind, 2 x 2 intervention trial was conducted in 5-14 y-old South African children (n = 404), who, on average, had mild-to-moderate VAD and ID. At baseline and after 3 mo, children received 1) iodine (191 mg I as oral iodized oil) + placebo (IS group), 2) VA (200000 IU VA as retinyl palmitate) + placebo (VAS group), 3) both iodine and VA (IS+VAS group), or 4) placebo. At baseline, 3 mo, and 6 mo, urinary iodine (UI), thyroid volume, thyrotropin (thyroid-stimulating hormone; TSH), total thyroxine (TT(4)), thyroglobulin, serum retinol (SR), and retinol-binding protein (RBP) were measured. RESULTS: SR and RBP increased significantly with VA supplementation (P < 0.05). For UI, SR, and RBP, there were no significant treatment interactions between iodine and vitamin A. The 3-factor and all three 2 factor interactions were significant for thyroid volume, TSH, and thyroglobulin (P < 0.001), whereas none of these interactions were significant for TT(4). There was a clear effect of VAS without IS on TSH, thyroglobulin, and thyroid volume; all 3 variables decreased significantly (P < 0.05). CONCLUSIONS: Iodine prophylaxis is effective in controlling ID in areas of poor vitamin A status. VA supplements are effective in treating VAD in areas of mild ID and have an additional benefit-through suppression of the pituitary TSHbeta gene, VAS can decrease excess TSH stimulation of the thyroid and thereby reduce the risk of goiter and its sequelae. PMID- 17921383 TI - One-time graded doses of vitamin A to weanling piglets enhance hepatic retinol but do not always prevent vitamin A deficiency. AB - BACKGROUND: Vitamin A supplements are administered to infants in developing countries at immunization contacts; doses of 50000 IU vitamin A are recommended. Doses of 100000 IU are given to children aged 0.5-1 y. The efficacy of these doses has not been adequately determined. OBJECTIVE: We aimed to quantify liver vitamin A after the administration of vitamin A doses to piglets. Piglets are a good model for infants because of their similar size, gastrointestinal anatomy, and vitamin A requirements. DESIGN: Castrated male piglets born to sows fed a vitamin A-depleted diet throughout 1 (parity A) or 3 (parity B) pregnancy and lactation cycles were randomly assigned to receive 1 of 4 oral vitamin A doses (ie, 0, 25000, 50000, or 100000 IU) at weaning (days 9-14). A vitamin A-depleted diet was fed until the piglets were killed on day 10. Serum retinol was measured on days 1, 2, 4, 7, and 10. The modified relative dose response was measured before supplementation and at the time of killing, and liver vitamin A concentration was measured. RESULTS: In both parities, 25000 IU did not result in a mean liver retinol reserve > 0.07 micromol/g liver (the deficiency cutoff). The 50000-IU dose increased mean reserves above 0.07 micromol/g only in parity A. Liver vitamin A reserves with the 100000-IU treatment were only 5% above those with the 50000-IU treatment. The modified relative dose-response test reflected differences in liver vitamin A stores in parity B, and the 0-IU group differed significantly from the 100000-IU group (P = 0.011). CONCLUSION: This piglet model suggests that, for supplementation to infants <6 mo old, a 50000-IU dose is likely to be more efficacious in mitigating deficiency than is a 25000-IU dose. PMID- 17921384 TI - Calcium requirements: new estimations for men and women by cross-sectional statistical analyses of calcium balance data from metabolic studies. AB - BACKGROUND: Low intakes of calcium are associated with an increased risk of both osteoporosis and cardiovascular disease. OBJECTIVE: To provide new estimates of the average calcium requirement for men and women, we determined the dietary calcium intake required to maintain neutral calcium balance. DESIGN: Calcium balance data [calcium intake -(fecal calcium + urinary calcium)] were collected from 155 subjects [women: n = 73; weight: 77.1 +/- 18.5 kg; age: 47.0 +/- 18.5 y (range: 20-75 y); men: n = 82; weight: 76.6 +/- 12.5 kg; age: 28.2 +/- 7.7 y (range: 19-64 y)] who participated in 19 feeding studies conducted in a metabolic unit. Balance data from the final 6-12 d of each dietary period (minimum length:18 d) of each study (1-9 observations per subject) were analyzed. Data were excluded if individual intakes of magnesium, copper, iron, phosphorus, or zinc fell below the estimated average requirements or exceeded the 99 th percentile of usual intakes from the 1994 Continuing Survey of Food Intakes by Individuals (for iron, above the upper limit). Daily intakes of calcium ranged between 415 and 1740 mg. The relation between intake and output was examined by fitting random coefficient models. Coefficients were included to test for sex and age differences. RESULTS: The models predicted a neutral calcium balance [defined as calcium output (Y) equal to calcium intake (C)] at intakes of 741 mg/d [95% prediction interval (PI): 507, 1035; Y = 148.29 + 0.80C], 9.4 mg kg body wt(-1) d(-1) [95% PI: 6.4, 12.9; Y = 1.44 + 0.85C], or 0.28 mg kcal(-1) d(-1) [95% PI: 0.19, 0.38; Y = 0.051 + 0.816C]. Neither age nor sex affected the estimates when calcium intakes were expressed as mg/d or as mg kg body wt(-1) d(-1). CONCLUSION: The findings suggest that the calcium requirement for men and women is lower than previously estimated. PMID- 17921385 TI - Methylation of estrogen receptor alpha and mutL homolog 1 in normal colonic mucosa: association with folate and vitamin B-12 status in subjects with and without colorectal neoplasia. AB - BACKGROUND: Greater promoter methylation in some tumor-suppressor genes underlies most sporadic colorectal cancers and increases with age in the colon. OBJECTIVE: We tested the hypothesis that biomarkers of folate and vitamin B-12 status are associated with estrogen receptor alpha (ERalpha) and mutL homolog 1 (MLH1) promoter methylation in subjects with and without neoplasia. DESIGN: Biopsies of normal-appearing colorectal mucosa from 156 subjects with and without colorectal neoplasia (disease free, n = 76; cancer, n = 28; adenoma, n = 35; hyperplastic polyps, n = 17) were obtained at colonoscopy and used to evaluate methylation in 7 CpG sites in the ERalpha promoter and 13 CpG sites in the MLH1 promoter. Blood samples were obtained for the measurement of serum and red cell folate, serum vitamin B-12, and plasma homocysteine concentrations. Methylation indexes were generated to reflect an average methylation value across all CpG dinucleotides in both ERalpha and MLH1. RESULTS: The methylation indexes for ERalpha and MLH1 generally were significantly (P < 0.05) higher in subjects with neoplasia than in disease-free subjects. The ERalpha methylation index correlated negatively with serum vitamin B-12 (r = -0.239, P = 0.003) and positively with plasma homocysteine (r = 0.188, P = 0.021). Disease status (P < 0.005), age (P < 0.001), and serum vitamin B-12 concentrations (P = 0.006) were independent determinants of ERalpha promoter methylation. Serum and red cell folate concentrations had no influence on ERalpha promoter methylation. CONCLUSION: Serum vitamin B-12 but not folate status may be associated with ERalpha promoter methylation in normal appearing colorectal mucosa. PMID- 17921387 TI - Effect of a 12-mo micronutrient intervention on learning and memory in well nourished and marginally nourished school-aged children: 2 parallel, randomized, placebo-controlled studies in Australia and Indonesia. AB - BACKGROUND: Little is known about the combined effect of micronutrients and essential fatty acids on cognitive function in school-aged children. OBJECTIVE: We assessed the effect of micronutrients, long-chain n-3 fatty acids, or both on indicators of cognitive performance in well-nourished and marginally nourished school-aged children. DESIGN: Two 2-by-2 factorial randomized controlled double blind trials were performed home-based in Adelaide, South Australia, and at 6 primary schools in Jakarta, Indonesia. A total of 396 children (aged 6-10 y) in Australia and 384 children in Indonesia were randomly allocated to receive a drink with a micronutrient mix (iron, zinc, folate, and vitamins A, B-6, B-12, and C), with docosahexanoic acid (DHA, 88 mg/d) and eicosapentaenoic acid (EPA, 22 mg/d), or with both or placebo 6 d/wk for 12 mo. Biochemical indicators were determined at baseline and 12 mo. Cognitive performance was measured at baseline, 6 mo, and 12 mo. RESULTS: The micronutrient treatment significantly improved plasma micronutrient concentrations in Australian and Indonesian children. DHA+EPA treatment increased plasma DHA and total plasma n-3 fatty acids in both countries. The micronutrient treatment resulted in significant increases in scores on tests representing verbal learning and memory in Australia (estimated effect size: 0.23; 95% CI: 0.01, 0.46). A similar effect was observed among Indonesian girls (estimated effect size: 0.32; 95% CI: -0.01, 0.64). No effects were found on tests measuring general intelligence or attention. No effects of DHA+EPA on the factors of cognitive tests were observed. CONCLUSION: In well nourished school-aged children, fortification with multiple micronutrients can result in improvements in verbal learning and memory. PMID- 17921386 TI - The association between betaine and choline intakes and the plasma concentrations of homocysteine in women. AB - BACKGROUND: Elevated total homocysteine (tHcy), a risk factor for many chronic diseases, can be remethylated to methionine by folate. Alternatively, tHcy can be metabolized by other 1-carbon nutrients, ie, betaine and its precursor, choline. OBJECTIVE: We aimed to assess the association between the dietary intakes of betaine and choline and the concentration of tHcy. DESIGN: We conducted a cross sectional analysis in 1477 women by using linear regression models to predict mean fasting tHcy by intakes of of betaine and choline. RESULTS: tHcy was 8% lower in the highest quintile of total betaine + choline intake than in the lowest quintile, even after control for folate intake (P for trend = 0.07). Neither choline nor betaine intake individually was significantly associated with tHcy. Choline from 2 choline-containing compounds, glycerophosphocholine and phosphocholine, was inversely associated with tHcy. These inverse associations were more pronounced in women with folate intake < 400 mug/d than in those with intakes >or=400 microg/d (P for interaction = 0.03 for phosphocholine) and in moderate alcohol drinkers (>or=15 g/d) than in nondrinkers or light drinkers (<15 g/d) (P for interaction = 0.02 for glycerophosphocholine and 0.04 for phosphocholine). The strongest dose response was seen in women with a low-methyl diet (high alcohol and low folate intake) (P for interaction = 0.002 for glycerophosphocholine and 0.001 for phosphocholine). CONCLUSIONS: Total choline + betaine intake was inversely associated with tHcy, as was choline from 2 water soluble choline-containing compounds. Remethylation of tHcy may be more dependent on the betaine pathway when methyl sources are low as a result of either inadequate folate intake or heavier alcohol consumption. PMID- 17921388 TI - Fortified complementary foods with or without alpha-amylase treatment increase hemoglobin but do not reduce breast milk intake of 9-mo-old Zambian infants. AB - BACKGROUND: Malnutrition in late infancy in developing countries may result from poor-quality complementary foods that displace breast milk. OBJECTIVE: The objective of the study was to assess the effects of fortified complementary blends of different energy densities on growth, hemoglobin concentrations, and breast milk intake of 9-mo-old Zambian infants. DESIGN: Infants were randomly assigned at 6 mo of age to receive for 3 mo a fortified blend of maize, beans, bambaranuts, and groundnuts [Chilenje Baby Mix (CBM); energy density: 68 kcal/100 g; n = 37] or a similar blend with alpha-amylase (CBMA; energy density: 106 kcal/100 g; n = 44). Cross-sectional data were obtained at 9 mo for a control group of infants (n = 69) not given the diets. Breast milk intake was measured by using the dose-to-the-mother deuterium dilution technique. RESULTS: No differences in weight or length z scores, all of which were within normal ranges, were seen between groups at 9 mo. Percentage fat mass was significantly (P = 0.01) greater in the infants in both the CBM (23.2 +/- 2.7%) and CBMA (23.4 +/- 2.5%) groups than in the control group (21.6 +/- 2.6%). Hemoglobin concentrations were significantly (P = 0.03) greater in both intervention groups (CBM group: 104 +/- 12 g/L: CBMA group: 103 +/- 12 g/L) than in the control group (98 +/- 14 g/L). Breast milk intake was not significantly (P = 0.87) different between groups (CBM group: 614 +/- 271 g/d; CBMA group: 635 +/- 193 g/d; control group: 653 +/- 221 g/d). CONCLUSIONS: The study foods improved hemoglobin concentrations without reducing breast milk intake and may be used to improve the nutritional status of infants in developing countries. PMID- 17921389 TI - Milk consumption during pregnancy is associated with increased infant size at birth: prospective cohort study. AB - BACKGROUND: Cow milk contains many potentially growth-promoting factors. OBJECTIVE: The objective was to examine whether milk consumption during pregnancy is associated with greater infant size at birth. DESIGN: During 1996-2002, the Danish National Birth Cohort collected data on midpregnancy diet through questionnaires and on covariates through telephone interviews and ascertained birth outcomes through registry linkages. Findings were adjusted for mother's parity, age, height, prepregnant BMI, gestational weight gain, smoking status, and total energy intake; father's height; and family's socioeconomic status The analyses included data from 50,117 mother-infant pairs. RESULTS: Mean (+/-SD) consumption of milk was 3.1 +/- 2.0 glasses/d. Milk consumption was inversely associated with the risk of small-for gestational age (SGA) birth and directly with both large-for-gestational age (LGA) birth and mean birth weight (P for trend < 0.001). In a comparison of women drinking >or=6 glasses/d with those drinking 0 glasses/d, the odds ratio for SGA was 0.51 (95% CI: 0.39, 0.65) and for LGA was 1.59 (1.16, 2.16); the increment in mean birth weight was 108 g (74, 143 g). We also found graded relations (P < 0.001) for abdominal circumference (0.52 cm; 0.35, 0.69 cm), placental weight (26 g; 15, 38 g), birth length (increment: 0.31 cm; 0.15, 0.46 cm), and head circumference (0.13 cm; 0.04, 0.25 cm). Birth weight was related to intake of protein, but not of fat, derived from milk. CONCLUSION: Milk intake in pregnancy was associated with higher birth weight for gestational age, lower risk of SGA, and higher risk of LGA. PMID- 17921390 TI - Longitudinal measures of circulating leptin and ghrelin concentrations are associated with the growth of young Peruvian children but are not affected by zinc supplementation. AB - BACKGROUND: Leptin, ghrelin, and insulin are hormonal regulators of energy balance and, therefore, may be related to growth during infancy. Zinc is essential for growth, and its growth effects may be mediated through these hormones. OBJECTIVE: We examined the effects of supplemental zinc on plasma leptin, ghrelin, and insulin concentrations among young children at risk of zinc deficiency and examined the relations between these hormones and physical growth. DESIGN: Children (n = 142) aged 6-8 mo were randomly assigned to receive 3 mg Zn/d as a supplement, in a fortified food, or as a placebo for 6 mo. Relations between hormones and anthropometric z scores, body composition, and growth rates were examined at baseline and 3 and 6 mo after the start of the intervention. RESULTS: No treatment group-related differences were found in plasma leptin, ghrelin, or glucose concentrations or in anthropometric z scores at 3 or 6 mo after the start of the zinc intervention. Neither plasma leptin nor ghrelin concentrations at baseline or 3 mo were predictive of subsequent changes in growth. However, changes in weight-for-age z scores over the two 3-mo time intervals were positively associated with subsequent leptin concentrations and inversely associated with subsequent plasma ghrelin concentrations. CONCLUSIONS: Supplemental zinc did not affect the children's growth, anthropometric indexes, or plasma hormone concentrations in this study population. Our results suggest that plasma leptin and ghrelin concentrations in later infancy are a consequence of previous weight changes rather than predictors of short-term growth. PMID- 17921392 TI - Effect of early infant feeding practices on infection-specific neonatal mortality: an investigation of the causal links with observational data from rural Ghana. AB - BACKGROUND: Strong associations between delayed initiation of breastfeeding and increased neonatal mortality (2-28 d) were recently reported in rural Ghana. Investigation into the biological plausibility of this relation and potential causal pathways is needed. OBJECTIVE: The objective was to assess the effect of early infant feeding practices (delayed initiation, prelacteal feeding, established neonatal breastfeeding) on infection-specific neonatal mortality in breastfed neonates aged 2-28 d. DESIGN: This prospective observational cohort study was based on 10 942 breastfed singleton neonates born between 1 July 2003 and 30 June 2004, who survived to day 2, and whose mothers were visited in the neonatal period. Verbal autopsies were used to ascertain the cause of death. RESULTS: One hundred forty neonates died from day 2 to day 28; 93 died of infection and 47 of noninfectious causes. The risk of death as a result of infection increased with increasing delay in initiation of breastfeeding from 1 h to day 7; overall late initiation (after day 1) was associated with a 2.6-fold risk [adjusted odds ratio (adj OR): 2.61; 95% CI: 1.68, 4.04]. Partial breastfeeding was associated with a 5.7-fold adjusted risk of death as a result of infectious disease (adj OR: 5.73; 95% CI: 2.75, 11.91). No obvious associations were observed between these feeding practices and noninfection specific mortality. Prelacteal feeding was not associated with infection (adj OR: 1.11; 95% CI: 0.66, 1.86) or noninfection-specific (adj OR: 1.33; 95% CI: 0.55, 3.22) mortality. CONCLUSIONS: This study provides the first epidemiologic evidence of a causal association between early breastfeeding and reduced infection-specific neonatal mortality in young human infants. PMID- 17921391 TI - Cysteine: a conditionally essential amino acid in low-birth-weight preterm infants? AB - BACKGROUND: Cyst(e)ine can be synthesized de novo from methionine and serine and is, therefore, a nonessential amino acid in human adults. Several studies have suggested that cyst(e)ine might be a conditionally essential amino acid in preterm infants because of biochemical immaturity. No data are available on cyst(e)ine requirements in low-birth-weight (LBW) preterm infants. OBJECTIVE: The aim was to determine cyst(e)ine requirements in LBW infants with gestational ages from 32 to 34 wk, measured 1 mo after birth with the use of the indicator amino acid oxidation technique. DESIGN: LBW infants were randomly assigned to 1 or 2 of the 5 formulas containing graded cystine concentrations (11, 22, 32, 43, or 65 mg cyst(e)ine/100 mL) and generous amounts of methionine. After 24-h adaptation, cyst(e)ine requirement was determined by (13)CO(2) release from [1 (13)C]phenylalanine in expired breath. (13)CO(2) enrichment was measured by isotopic ratio mass spectrometry. RESULTS: Cyst(e)ine requirement was determined in 25 LBW infants with a mean (+/-SD) gestational age of 33 +/- 1 wk and birth weight of 1.78 +/- 0.32 kg. Fractional oxidation of [1-(13)C]phenylalanine did not differ between the 5 groups. CONCLUSIONS: There is no evidence for limited endogenous cyst(e)ine synthesis in 4-wk-old LBW preterm infants born at gestational ages from 32 to 34 wk. It is safe to conclude that the cyst(e)ine requirement is <18 mg kg(-1) d(-1) providing generous amounts of methionine and that cyst(e)ine is probably not a conditionally essential amino acid in fully enterally fed LBW preterm infants born at 32-34 wk. PMID- 17921393 TI - The gut takes nearly all: threonine kinetics in infants. AB - BACKGROUND: Threonine is an essential amino acid that is abundantly present in intestinally produced glycoproteins. Animal studies show that intestinal first pass threonine metabolism is high, particularly during a restricted enteral protein intake. OBJECTIVE: The objective of the study was to quantify intestinal first-pass threonine metabolism in preterm infants during full enteral feeding and during restricted enteral intake. DESIGN: Eight preterm infants (x +/- SD birth weight: 1.1 +/- 0.1 kg; gestational age: 29 +/- 2 wk) were studied during 2 periods. During period A, 40% of total intake was administered enterally and 60% was administered parenterally. Total threonine intake was 58 +/- 6 micromol kg( 1) h(-1). During period B, the infants received full enteral feeding, and the total threonine intake was 63 +/- 6 micromol kg(-1) h(-1). Dual stable-isotope tracer techniques were used to assess splanchnic and whole-body threonine kinetics. RESULTS: The fractional first-pass threonine uptake by the intestine was remarkably high in both periods: 82 +/- 6% during partial enteral feeding and 70 +/- 6% during full enteral feeding. Net threonine retention was not affected by the route of feeding. CONCLUSION: In preterm infants, the splanchnic tissues extract a very large amount of the dietary threonine intake, which indicates a high obligatory visceral need for threonine, presumably for the purposes of synthesis. PMID- 17921395 TI - Excretion of starch and esterified short-chain fatty acids by ileostomy subjects after the ingestion of acylated starches. AB - BACKGROUND: Short-chain fatty acids (SCFAs) have a role in maintaining bowel health and can assist in the prevention and treatment of colonic disease. The ability of acylated starches to deliver SCFAs to the large bowel has been shown in animal studies but has not been established in humans. OBJECTIVE: The aim was to determine whether cooked, highly acylated starches were resistant to small intestinal digestion in ileostomy volunteers. DESIGN: Volunteers consumed single doses of custards containing 20 g cooked acetylated, propionylated, or butyrylated high-amylose maize starches (HAMSA, HAMSP, and HAMSB, respectively) on each collection day. The amounts of starch and of esterified SCFAs ingested and subsequently excreted in the stoma effluent were measured. Custards containing unacylated high-amylose maize starch (Hylon VII, HAMS) and low-amylose maize starch (3401C, LAMS) were consumed as controls. RESULTS: Between 73% and 76% of the esterified SCFAs survived small intestinal digestion, which showed the potential of acylated starches to deliver specific SCFAs to the large bowel. The resistance of starches to small intestinal digestion as measured by ileal excretion was significantly greater for HAMSA, HAMSP, HAMSB, and HAMS than for LAMS (P < 0.001). The concentration of acetate in stoma digesta was higher than expected in all groups; this additional acid may have been derived from endogenous sources. CONCLUSIONS: Acylated starches are a potentially effective method of delivering significant quantities of specific SCFAs to the colon in humans. These products have potential application in the treatment and prevention of bowel disorders amenable to modulation by SCFAs. PMID- 17921394 TI - Relation between prenatal lipid-soluble micronutrient status, environmental pollutant exposure, and birth outcomes. AB - BACKGROUND: Adverse postnatal health effects have been associated with compromised fetal growth, which makes it essential to understand its determinants. Significant effects of environmental pollutants on birth outcomes have been observed in our study population, and nutritional status may be an additional factor influencing fetal development and effects of environmental toxins. OBJECTIVE: The objective of the study was to examine the relations between birth outcomes and lipid-soluble plasma micronutrient concentrations and to explore interactions between micronutrients and environmental pollutant exposure in newborns in Krakow, Poland. DESIGN: In this prospective cohort study, retinol, alpha-tocopherol, and carotenoids were measured in maternal and cord blood samples obtained at delivery (251 maternal-newborn pairs), and birth weight, birth length, head circumference (HC), and gestational age were evaluated. Linear regression analysis was used to estimate the effects of micronutrients while covariates were controlled for. Interaction terms assessed whether the effects of polycyclic aromatic hydrocarbons (PAHs), common environmental pollutants, varied by nutrient status. RESULTS: Infants whose mothers had low plasma alpha-tocopherol concentrations (below the median) weighed 92.9 g less and had 0.41-cm smaller HCs than did infants whose mothers had high alpha-tocopherol concentrations. Infants with low plasma retinol (below the median) weighed 125.9 g less and had 0.31-cm smaller HCs. There was no evidence of an interaction between PAHs and micronutrients, although power was limited. CONCLUSION: Maternal alpha-tocopherol and cord retinol concentrations were significantly and positively associated with BW and HC. These micronutrients may have direct effects or may be markers for other underlying determinants of these pregnancy outcomes. PMID- 17921396 TI - Effect of high-dose vitamin A supplementation on the immune response to Bacille Calmette-Guerin vaccine. AB - BACKGROUND: Vitamin A supplementation (VAS) at birth has been associated with decreased mortality in Asia. Bacille Calmette-Guerin (BCG) vaccine is given at birth in tuberculosis-endemic countries. Previous studies suggest that VAS may influence the immune response to vaccines. OBJECTIVE: Our objective was to examine whether VAS influences the immune response to simultaneously administered BCG vaccine. DESIGN: Within a randomized trial of 50,000 IU vitamin A or placebo given with BCG vaccine at birth in Guinea-Bissau, 2710 infants were examined for BCG scar formation and delayed-type hypersensitivity (DTH) to purified protein derivative of Mycobacterium tuberculosis (PPD) at 2 and 6 mo of age. The ex vivo cytokine response to PPD was measured in 607 infants. RESULTS: At 2 mo of age, 39% (43% of the boys and 34% of the girls) responded to PPD. The prevalence ratio of a measurable PPD reaction for VAS compared with placebo recipients was 0.90 (95% CI: 0.80, 1.02) for all infants, 0.81 (95% CI: 0.69, 0.95) for boys, and 1.04 (95% CI: 0.86, 1.26) for girls. At 6 mo of age, 42% of the infants responded to PPD. No difference was observed between VAS and placebo recipients. The prevalence of BCG scar was not affected by VAS. The ex vivo interferon-gamma response to PPD was increased by VAS (means ratio: 1.40; 95% CI: 1.03, 1.91). CONCLUSIONS: VAS with BCG vaccination does not appear to interfere with the long term immune response to BCG. However, VAS temporarily altered the DTH reaction to PPD in boys at 2 mo of age, suggesting sex differences in the immunologic response to VAS given with BCG. This trial was registered at www.clinicaltrials.gov as #NCT00168597. PMID- 17921397 TI - Dietary glycemic index, glycemic load, and the risk of breast cancer in an Italian prospective cohort study. AB - BACKGROUND: Interest in the roles of glycemic index (GI) and glycemic load (GL) in breast cancer etiology has been stimulated by indications that disease risk is linked to insulinemia, sex hormone bioavailability, and insulin-like growth factor 1. OBJECTIVE: We aimed to determine whether GI and GL were associated with the risk of breast cancer in a cohort of Italian women volunteers from Northern Italy, who enrolled between 1987-1992 in the Hormones and Diet in the Etiology of Breast Tumors Study (ORDET Study). DESIGN: Volunteers completed a semiquantitative food-frequency questionnaire, and anthropometric and lifestyle data were collected. Dietary GI and GL in relation to breast cancer risk were examined in 8926 cohort women, including 289 with breast cancer identified after a mean follow-up of 11.5 y. RESULTS: The relative risk (RR) of breast cancer in the highest (versus lowest) quintiles of GI and GL was 1.57 (95% CI: 1.04, 2.36; P for trend = 0.040) and 2.53 (95% CI: 1.54, 4.16; P for trend = 0.001), respectively. Total carbohydrate intake was not associated with greater breast cancer risk, but high carbohydrate from high-GI foods was. When women were categorized by baseline menopausal status and body mass index (BMI; in kg/m(2)), the increased risk of dietary GL was confined to those who were premenopausal (RR = 3.89; 95% CI: 1.81, 8.34) and who had normal BMI (ie, <25) (RR = 5.79; 95% CI: 2.60, 12.90) (P for trend = 0.001 for both). CONCLUSIONS: A high-GL diet may increase the risk of breast cancer in Italian women. The effect is particularly evident in premenopausal women and those with BMI < 25. PMID- 17921399 TI - Fructose intake is a predictor of LDL particle size in overweight schoolchildren. AB - BACKGROUND: High amounts of dietary fructose may contribute to dyslipidemia in adults, but there are few data in children. Childhood adiposity is associated with smaller LDL particle size, but the dietary predictors of LDL size in overweight children have not been studied. OBJECTIVES: We aimed to determine whether LDL particle size is associated with dietary factors and specifically with fructose intake in normal-weight and overweight children. DESIGN: In a cross sectional study of normal-weight and overweight 6-14 y-old Swiss children (n = 74), dietary intakes were assessed by using two 24-h-recalls and a 1-d dietary record. Body mass index (BMI) and waist-hip ratio (WHR) were measured, and plasma lipid profile and LDL particle size were determined. RESULTS: Compared with the normal-weight group, overweight children had significantly higher plasma triacylglycerol concentrations, lower HDL-cholesterol concentrations, and smaller LDL particle size (P < 0.05). LDL particle size was inversely correlated to BMI SD scores and WHR (P = 0.007). Although there were no significant differences in total fructose intake, the overweight children consumed a significantly (P < 0.05) higher percentage of fructose from sweets and sweetened drinks than did the normal-weight children. After control for adiposity, the only dietary factor that was a significant predictor of LDL particle size was total fructose intake (P = 0.024). CONCLUSIONS: In school-age children, greater total and central adiposity are associated with smaller LDL particle size and lower HDL cholesterol. Overweight children consume more fructose from sweets and sweetened drinks than do normal-weight children, and higher fructose intake predicts smaller LDL particle size. PMID- 17921400 TI - Genomic methylation of peripheral blood leukocyte DNA: influences of arsenic and folate in Bangladeshi adults. AB - BACKGROUND: Studies in cell culture and animal models indicate that arsenic exposure induces modifications in DNA methylation, including genome-wide DNA hypomethylation. It is not known whether arsenic exposure influences genomic DNA methylation in human populations chronically exposed to arsenic-contaminated drinking water. OBJECTIVE: The objective of this study was to determine whether arsenic is associated with genomic hypomethylation of peripheral blood leukocyte (PBL) DNA in Bangladeshi adults who are chronically exposed to arsenic. We also investigated whether arsenic-induced alterations in DNA methylation may be influenced by folate nutritional status. DESIGN: PBL DNA methylation and concentrations of plasma folate, plasma arsenic, and urinary arsenic were assessed in 294 adults in Araihazar, Bangladesh. Genomic PBL DNA methylation was measured by using a [(3)H]-methyl incorporation assay. RESULTS: Urinary arsenic, plasma arsenic, and plasma folate were positively associated with the methylation of PBL DNA (P = 0.009, 0.03, and 0.03, respectively). Stratification of participants by folate nutritional status [<9 nmol/L (n = 190) or >or=9 nmol/L (n = 104)] showed that the associations between arsenic exposure and methylation of PBL DNA were restricted to persons with folate concentrations >or= 9 nmol/L. CONCLUSIONS: Contrary to our a priori hypothesis, arsenic exposure was positively associated with genomic PBL DNA methylation in a dose-dependent manner. This effect is modified by folate, which suggests that arsenic-induced increases in DNA methylation cannot occur in the absence of adequate folate. The underlying mechanisms and physiologic implications of increased genomic DNA methylation are unclear, and they warrant further study. PMID- 17921398 TI - Serum zinc and pneumonia in nursing home elderly. AB - BACKGROUND: Zinc plays an important role in immune function. The association between serum zinc and pneumonia in the elderly has not been studied. OBJECTIVE: The objective was to determine whether serum zinc concentrations in nursing home elderly are associated with the incidence and duration of pneumonia, total and duration of antibiotic use, and pneumonia-associated and all-cause mortality. DESIGN: This observational study was conducted in residents from 33 nursing homes in Boston, MA, who participated in a 1-y randomized, double-blind, and placebo controlled vitamin E supplementation trial; all were given daily doses of 50% of the recommended dietary allowance of essential vitamins and minerals, including zinc. Participants with baseline (n = 578) or final (n = 420) serum zinc concentrations were categorized as having low (<70 microg/dL) or normal (>or=70 microg/dL) serum zinc concentrations. Outcome measures included the incidence and number of days with pneumonia, number of new antibiotic prescriptions, days of antibiotic use, death due to pneumonia, and all-cause mortality. RESULTS: Compared with subjects with low zinc concentrations, subjects with normal final serum zinc concentrations had a lower incidence of pneumonia, fewer (by almost 50%) new antibiotic prescriptions, a shorter duration of pneumonia, and fewer days of antibiotic use (3.9 d compared with 2.6 d) (P or=19 y with low serum vitamin B-12 (<258 pmol/L) and mean corpuscular volume (MCV) measured between 1995 and 2004 were identified from medical records. The proportion and odds ratios of individuals with low serum vitamin B-12 without macrocytosis by sex, race, and age according to prefortification (n = 86), perifortification (n = 138), and postfortification (n = 409) periods were determined. RESULTS: MCV was significantly lower in the postfortification period (88.6 fL) than in the prefortification (94.4 fL; P < 0.001) and perifortification (90.6 fL; P = 0.007) periods. The proportion of subjects with low serum vitamin B 12 without macrocytosis was significantly higher in the postfortification (approximately 87%) and perifortification (approximately 85%) periods than in the prefortification period (approximately 70%; P < 0.001). In a sex-, race-, and age adjusted analysis, the odds ratio for having low serum vitamin B-12 without macrocytosis was 3.0 (95% CI: 1.7, 5.2) in the postfortification period relative to the prefortification period. CONCLUSIONS: Subjects with low serum vitamin B-12 were likely to be without macrocytosis in the postfortification period. MCV should not be used as a marker for vitamin B-12 insufficiency. It is possible that folic acid fortification may have led to a correction of macrocytosis associated with vitamin B-12 insufficiency. PMID- 17921402 TI - The 2005 Dietary Guidelines for Americans and risk of the metabolic syndrome. AB - BACKGROUND: The 2005 Dietary Guidelines for Americans Index (DGAI) was created to assess adherence to the dietary recommendations of the 2005 Dietary Guidelines for Americans (DGA) in relation to chronic disease risk. OBJECTIVE: The objective was to assess the relation between dietary patterns consistent with the 2005 DGA as measured by the DGAI and both the prevalence of the metabolic syndrome (MetS) and individual MetS risk factors. DESIGN: DGAI scores and metabolic risk factors for MetS were assessed in a cross-sectional study of 3177 participants from the Framingham Heart Study Offspring Cohort. MetS was defined on the basis of the National Cholesterol Education Program Adult Treatment Panel III criteria. RESULTS: After adjustment for potential confounders, the DGAI score was inversely related to waist circumference (P for trend < 0.001), triacylglycerol concentration (P for trend = 0.005), both diastolic (P for trend = 0.002) and systolic (P for trend = 0.01) blood pressure, the prevalence of abdominal adiposity (P for trend < 0.001), and hyperglycemia (P for trend = 0.03). The prevalence of MetS was significantly lower in individuals in the highest DGAI quintile category than in those in the lowest category (odds ratio: 0.64; 95% CI: 0.47, 0.88; P for trend = 0.005) when those being treated for any of the risk factors were excluded. There was a significant interaction between DGAI score and age; the association between the DGAI score and MetS was confined largely to adults younger than 55 y (odds ratio: 0.57; 95% CI: 0.36, 0.92; P for trend < 0.01). CONCLUSIONS: A dietary pattern consistent with the 2005 DGA was associated with a lower prevalence of MetS-a potential risk factor for CVD. PMID- 17921403 TI - Folic acid supplementation lowers blood arsenic. AB - BACKGROUND: Chronic arsenic exposure currently affects >100 million persons worldwide. Methylation of ingested inorganic arsenic (InAs) to monomethylarsonic (MMAs) and dimethylarsinic (DMAs) acids relies on folate-dependent one-carbon metabolism and facilitates urinary arsenic elimination. OBJECTIVE: We hypothesized that folic acid supplementation to arsenic-exposed Bangladeshi adults would increase arsenic methylation and thereby lower total blood arsenic. DESIGN: In this randomized, double-blind, placebo-controlled trial, we evaluated blood concentrations of total arsenic, InAs, MMAs, and DMAs in 130 participants with low plasma folate (<9 nmol/L) before and after 12 wk of supplementation with folic acid (400 microg/d) or placebo. RESULTS: MMAs in blood was reduced by a mean +/- SE of 22.24 +/- 2.86% in the folic acid supplementation group and by 1.24 +/- 3.59% in the placebe group (P < 0.0001). There was no change in DMAs in blood; DMAs is rapidly excreted in urine as evidenced by an increase in urinary DMAs (P = 0.0099). Total blood arsenic was reduced by 13.62% in the folic acid supplementation group and by 2.49% in the placebo group (P = 0.0199). CONCLUSIONS: Folic acid supplementation to participants with low plasma concentrations of folate lowered blood arsenic concentrations, primarily by decreasing blood MMAs and increasing urinary DMAs. Therapeutic strategies to facilitate arsenic methylation, particularly in populations with folate deficiency or hyperhomocysteinemia or both, may lower blood arsenic concentrations and thereby contribute to the prevention of arsenic-induced illnesses. PMID- 17921404 TI - Dietary carbohydrate and the progression of age-related macular degeneration: a prospective study from the Age-Related Eye Disease Study. AB - BACKGROUND: Cross-sectional studies indicate that diets that provide a higher dietary glycemic index (dGI) are associated with a greater risk of age-related macular degeneration (AMD). No prospective studies have addressed this issue. OBJECTIVE: The objective was to prospectively evaluate the effect of baseline dGI on the progression of AMD. DESIGN: dGI was calculated as the weighted average of GIs from foods and was evaluated as being above or below the sex median (women: 77.9; men: 79.3) for 3977 participants aged 55-80 y (58% women) in the Age Related Eye Disease Study. The 7232 eligible eyes without advanced AMD were classified into 1 of 3 AMD categories: group 1 (nonextensive small drusen), group 2 (intermediate drusen, extensive small drusen, or pigmentary abnormalities), or group 3 (large drusen or extensive intermediate drusen). With the use of multifailure Cox proportional-hazards regression, we modeled the time to the maximal progression to evaluate the relation between dGI and the risk of AMD. RESULTS: Overall, the multivariate-adjusted risk of progression over 8 y of follow-up (x: 5.4 y) was significantly higher (risk ratio: 1.10; 95% CI: 1.00, 1.20; P = 0.047) in the high-dGI group than in the low-dGI group. The risk of progression for groups 1, 2, and 3 eyes was 5%, 8%, and 17% greater, respectively (P for trend < 0.001). The latter gives an estimate that 7.8% of new advanced AMD cases would be prevented in 5 y if people consumed the low-dGI diet. CONCLUSION: Persons at risk of AMD progression, especially those at high risk of advanced AMD, may benefit from consuming a smaller amount of refined carbohydrates. PMID- 17921405 TI - The metabolic syndrome in adults prenatally exposed to the Dutch famine. AB - BACKGROUND: Epidemiologic studies have shown that the metabolic syndrome may originate in utero. OBJECTIVE: We aimed to determine whether exposure to prenatal famine is associated with a greater prevalence of the metabolic syndrome. DESIGN: We assessed the prevalence of the metabolic syndrome according to the National Cholesterol Education Program definition in 783 members of the Dutch Famine Birth Cohort. Participants were born as term singletons around the time of the 1944 1945 Dutch famine. RESULTS: Exposure to famine during gestation was not significantly associated with the metabolic syndrome (odds ratio: 1.2; 95% CI: 0.9, 1.7). Birth weight also was not significantly associated with the metabolic syndrome (odds ratio: 1.3/1-kg decrease in birth weight; 95% CI: 0.9, 1.8/1-kg decrease in birth weight). Exposure to famine during gestation was associated with significantly higher triacylglycerol concentrations (0.1 g/L; 0.0, 0.2 g/L). Men exposed to famine in early gestation had significantly lower HDL-cholesterol concentrations (-0.08 mmol/L; -0.14, 0.00 mmol/L) than did unexposed men. CONCLUSIONS: Prenatal exposure to famine or reduced birth weight is not associated with a significantly greater prevalence of the metabolic syndrome. Our findings suggest that, although elements of the metabolic syndrome may be programmed by fetal undernutrition, the origin of the syndrome as a whole is not likely to be found in poor nutrition during gestation. PMID- 17921406 TI - Dietary nutrient intakes and skin-aging appearance among middle-aged American women. AB - BACKGROUND: Nutritional factors play a key role in normal dermatologic functioning. However, little is known about the effects of diet on skin-aging appearance. OBJECTIVE: We evaluated the associations between nutrient intakes and skin-aging appearance. DESIGN: Using data from the first National Health and Nutrition Examination Survey, we examined associations between nutrient intakes and skin aging in 4025 women (40-74 y). Nutrients were estimated from a 24-h recall. Clinical examinations of the skin were conducted by dermatologists. Skin aging appearance was defined as having a wrinkled appearance, senile dryness, and skin atrophy. RESULTS: Higher vitamin C intakes were associated with a lower likelihood of a wrinkled appearance [odds ratio (OR) 0.89; 95% CI: 0.82, 0.96] and senile dryness (OR: 0.93; 95% CI: 0.87, 0.99). Higher linoleic acid intakes were associated with a lower likelihood of senile dryness (OR: 0.75; 95% CI: 0.64, 0.88) and skin atrophy (OR: 0.78; 95% CI 0.65, 0.95). A 17-g increase in fat and a 50-g increase in carbohydrate intakes increased the likelihood of a wrinkled appearance (OR: 1.28 and 1.36, respectively) and skin atrophy (OR: 1.37 and 1.33, respectively). These associations were independent of age, race, education, sunlight exposure, income, menopausal status, body mass index, supplement use, physical activity, and energy intake. CONCLUSIONS: Higher intakes of vitamin C and linoleic acid and lower intakes of fats and carbohydrates are associated with better skin-aging appearance. Promoting healthy dietary behaviors may have additional benefit for skin appearance in addition to other health outcomes in the population. PMID- 17921407 TI - Effect of the interaction between the fatty acid binding protein 2 gene Ala54Thr polymorphism and dietary fatty acids on peripheral insulin sensitivity: a cross sectional study. AB - BACKGROUND: The intestinal fatty acid-binding protein (FABP2) is involved in the intracellular transport and metabolism of fatty acids and may affect insulin sensitivity and glucose metabolism. OBJECTIVE: The objective was to study the effect of interaction between the Ala54Thr polymorphism of the FABP2 gene (FABP2) and the type of dietary cooking oil used on peripheral insulin sensitivity in a population from southern Spain. DESIGN: The study was cross-sectional. Anthropometric measurements were obtained for 1226 persons aged 18-65 y selected randomly from the municipal census of Pizarra, Spain. An oral-glucose-tolerance test was given to 1020 of these persons. Insulin resistance was measured by homeostasis model assessment. Samples of the cooking oil being used were taken from the kitchens of a random subset of 538 persons. RESULTS: Persons who consumed sunflower oil and who also had the Thr54 variant had higher insulin resistance than did those who consumed olive oil (P = 0.01). We detected an interaction between the Ala54Thr polymorphism and the type of oil consumed that accounted for the variance in insulin resistance (P = 0.02). CONCLUSIONS: The effect of dietary fatty acids on the populational pattern of insulin resistance is not independent of the Ala54Thr polymorphism of FABP2. An interaction existed between this polymorphism and the intake of dietary fats in a population with a high intake of monounsaturated fatty acids. PMID- 17921408 TI - Ala12 variant of the peroxisome proliferator-activated receptor-gamma gene (PPARG) is associated with higher polyunsaturated fat in adipose tissue and attenuates the protective effect of polyunsaturated fat intake on the risk of myocardial infarction. AB - BACKGROUND: Intake of polyunsaturated fat is protective against the development of coronary heart disease. Less is known about the genetic variation modulating this association. The Ala12 allele of the peroxisome proliferator-activated receptor-gamma gene (PPARG) decreases the lipolysis of triacylglycerols in adipose tissue, which results in the accumulation of fatty acids in adipocytes. OBJECTIVE: We aimed to determine whether the Pro12Ala polymorphism interacts with polyunsaturated fat intake to affect the risk of myocardial infarction (MI). DESIGN: Cases (n = 1805) with a first nonfatal acute MI and population-based controls matched by age, sex, and area of residence (n = 1805) living in Costa Rica were genotyped for the PPARG Pro12Ala genetic polymorphism. Polyunsaturated fat intake was determined by use of a validated food-frequency questionnaire and by gas chromatography analysis of adipose tissue. Odds ratios and 95% CIs for MI were estimated by use of logistic regression. RESULTS: The relative allele frequencies of the Ala12 allele were 10% in controls and 11% in cases. Odds ratios (95% CI) for MI per each 5% increase in energy from polyunsaturated fat were 0.66 (0.53, 0.82) in Pro12/Pro12 subjects and 0.93 (0.61, 1.42) in carriers of the Ala12 allele (P for interaction = 0.03). Increments (95% CI) of polyunsaturated fat in adipose tissue per 5% increment in dietary intake were 5.4% (4.9%, 5.9%) in Pro12/Pro12 homozygotes, 6.9% (6.0%, 7.9%) in Pro12/Ala12 heterozygotes, and 7.7% (3.2%, 12.2%) in Ala12/Ala12 homozygotes (P for interaction = 0.016). CONCLUSIONS: The protective effect of polyunsaturated fat intake on MI is attenuated in carriers of the Ala12 allele of PPARG. PMID- 17921409 TI - Tea drinking is associated with benefits on bone density in older women. AB - BACKGROUND: Impaired hip structure assessed by dual-energy X-ray absorptiometry (DXA) areal bone mineral density (aBMD) is an independent predictor for osteoporotic hip fracture. Some studies suggest that tea intake may protect against bone loss. OBJECTIVE: Using both cross-sectional and longitudinal study designs, we examined the relation of tea consumption with hip structure. DESIGN: Randomly selected women (n = 1500) aged 70-85 y participated in a 5-y prospective trial to evaluate whether oral calcium supplements prevent osteoporotic fractures. aBMD at the hip was measured at years 1 and 5 with DXA. A cross sectional analysis of 1027 of these women at 5 y assessed the relation of usual tea intake, measured by using a questionnaire, with aBMD. A prospective analysis of 164 women assessed the relation of tea intake at baseline, measured by using a 24-h dietary recall, with change in aBMD from years 1 to 5. RESULTS: In the cross sectional analysis, total hip aBMD was 2.8% greater in tea drinkers (x: 806; 95% CI: 797, 815 mg/cm(2)) than in non-tea drinkers (784; 764, 803 mg/cm(2)) (P < 0.05). In the prospective analysis over 4 y, tea drinkers lost an average of 1.6% of their total hip aBMD (-32; -45, -19 mg/cm(2)), but non-tea drinkers lost 4.0% (-13; -20, -5 mg/cm(2)) (P < 0.05). Adjustment for covariates did not influence the interpretation of results. CONCLUSION: Tea drinking is associated with preservation of hip structure in elderly women. This finding provides further evidence of the beneficial effects of tea consumption on the skeleton. PMID- 17921410 TI - Coffee intake and the incidence of hypertension. PMID- 17921411 TI - Effects of glycemic load on weight loss in overweight adults. PMID- 17921412 TI - Validation of energy intake during calorie restriction with the doubly labeled water method and changes in body composition. PMID- 17921413 TI - The effect of soybean-based foods on plasma lipid and lipoprotein concentrations. PMID- 17921414 TI - A controlled trial of reduced meal frequency without caloric restriction in healthy, normal-weight, middle-aged adults. PMID- 17921415 TI - Is there any monitoring of the quality of vitamin A capsules used in supplementation programs? PMID- 17921416 TI - Neuromuscular and lower limb biomechanical differences exist between male and female elite adolescent soccer players during an unanticipated side-cut maneuver. AB - BACKGROUND: Female athletes are 2 to 8 times more likely than male athletes to injure the anterior cruciate ligament during a non-contact athletic maneuver. Identifying anterior cruciate ligament injury risk factors in female athletes may help with the development of preventive training programs aimed at reducing injury rates. HYPOTHESIS: Differences between genders in lower limb kinematics, kinetics, and neuromuscular patterns will be identified in an adolescent soccer population during an unanticipated side-cut maneuver. STUDY DESIGN: Controlled laboratory study. METHODS: Forty-two elite adolescent soccer players (21 male and 21 female) performed an unanticipated side-cut maneuver, with the 3-dimensional kinematic, kinetic, and electromyographic lower limb data being analyzed using principal component analysis. RESULTS: The female athletes had higher gastrocnemius activity, normalized to maximal voluntary isometric contractions, and a mediolateral gastrocnemius activation imbalance that was not present in the male athletes during early stance to midstance of the side-cut. Female athletes demonstrated greater rectus femoris muscle activity throughout stance, and the only hamstring difference identified was a mediolateral activation imbalance in male athletes only. Female athletes performed the side-cut with less hip flexion and more hip external rotation and also generated a smaller hip flexion moment compared with the male athletes. CONCLUSION: This is the first study to identify gender-related differences in gastrocnemius muscle activity during an unanticipated cutting maneuver. CLINICAL RELEVANCE: The increased and imbalanced gastrocnemius muscle activity, combined with increased rectus femoris muscle activity and reduced hip flexion angles and moments in female subjects, may all have important contributing roles in the higher noncontact ACL injury rates observed in female athletes. PMID- 17921417 TI - Plasma glucose and the action of calorie restriction on aging. AB - We tested the hypothesis that retardation of aging by caloric restriction is due in part to decreased levels of plasma glucose over the life span. Male C57BL/6 mice expressing a human GLUT4 minigene (transgenic [TG] mice) and their nontransgenic littermates (NTG mice) were maintained under specific pathogen-free conditions. Mice were fed ad libitum (A mice) or 40% less than ad libitum (R mice) from age 6 weeks. Over the life span there were three different levels of plasma glucose, with NTGA mice having the highest daily levels, TGR mice the lowest daily values, and TGA and NTGR mice having similar levels intermediate between these values. Despite differences in plasma glucose, the differences measured in longevity (50% and 10% survival), physiology and tissue pathology were associated with diet rather than with levels of plasma glucose. We conclude that decreased plasma glucose over the life span is not an important factor in the action of calorie restriction on aging processes. PMID- 17921418 TI - Optimization of dietary restriction protocols in Drosophila. AB - Dietary restriction (DR) extends life span in many organisms, through unknown mechanisms that may or may not be evolutionarily conserved. Because different laboratories use different diets and techniques for implementing DR, the outcomes may not be strictly comparable. This complicates intra- and interspecific comparisons of the mechanisms of DR and is therefore central to the use of model organisms to research this topic. Drosophila melanogaster is an important model for the study of DR, but the nutritional content of its diet is typically poorly defined. We have compared fly diets composed of different yeasts for their effect on life span and fecundity. We found that only one diet was appropriate for DR experiments, indicating that much of the published work on fly "DR" may have included adverse effects of food composition. We propose procedures to ensure that diets are suitable for the study of DR in Drosophila. PMID- 17921419 TI - Lifelong caloric restriction and interleukin-6 secretion from adipose tissue: effects on physical performance decline in aged rats. AB - We investigated whether caloric restriction (CR) improves physical performance in a rodent model of aging, and whether this effect is accompanied with a decrease in visceral adipose tissue production of proinflammatory cytokines. Body composition, standardized physical performance measures, as well as in vitro visceral adipose tissue cytokine secretion and circulating levels of an inflammatory marker were cross-sectionally assessed in ad libitum (AL)-fed and lifelong CR Fischer 344 x Brown Norway male rats aged 18, 24, and 29 months. Fat to lean mass ratio increased and physical performance declined with age in the AL rats. Compared to AL rats, CR rats had lower fat mass, fat to lean ratio, adipose tissue secretion of interleukin-6, and circulating levels of C-reactive protein, and higher physical performance scores. Therefore, CR may be an effective intervention for improving functional status into advanced age and is perhaps mediated via a reduction in adipose tissue-generated proinflammatory cytokine production. PMID- 17921420 TI - Gene expression, fiber type, and strength are similar between left and right legs in older adults. AB - Methodological issues relevant to studies using microarrays and reverse transcription-polymerase chain reaction (RT-PCR) in human aging have rarely been evaluated. Because aging may accentuate biological differences between muscles, we compared transcriptome expression patterns, targeted messenger RNA (mRNA) abundance, strength, and muscle fiber type in the right and left legs of older adults. Muscle biopsies were taken from each Vastus lateralis in eight older (71 +/- 2 years) men, and isometric strength was determined. Samples were analyzed using an Affymetrix gene array, ATPase histochemistry, and RT-PCR for mRNA species involved in metabolism, apoptosis, vascular growth, and antioxidant status. Microarray analysis found that 31 of 5499 genes (0.6%) were significantly different between legs (negative log of the p value [NLOGP] >/= 2.0, but fold < 1.5), with only one gene, jumonji domain containing 1C (JMJD1C), being significantly different by >/= 1.50-fold. None of the mRNA species, or muscle fiber type, size, or strength, was different between legs. These findings are important for the design and analysis of studies using muscle data in older adults. PMID- 17921421 TI - A method for identifying biomarkers of aging and constructing an index of biological age in humans. AB - This study was conducted to identify biomarkers of aging and to construct an index of biological age in humans. Healthy adult men (n = 86) who had received an annual health examination from 1992 through 1998 were studied. From 29 physiological variables, five variables (forced expiratory volume in 1 second, systolic blood pressure, hematocrit, albumin, blood urea nitrogen) were selected as candidate biomarkers of aging. Five candidate biomarkers expressed substantial covariance along one principal component. The first principal component obtained from a principal component analysis was used to calculate biological age scores (BAS). Individual BAS showed high longitudinal stability of age-related changes. Age-related changes of BAS are characterized by three components: age, peak functional capacity, and aging rate. A logistic regression analysis suggested that aging rate was influenced by environmental factors, but peak functional capacity was almost independent of environmental factors. PMID- 17921423 TI - Driving and the promotion of safe mobility in older populations. PMID- 17921424 TI - A randomized trial of an education program to enhance older driver performance. AB - BACKGROUND: This study was designed to determine whether an education program consisting of classroom and on-road training could enhance driving performance. METHODS: This randomized controlled trial with blinded endpoint assessment enrolled 126 community-living drivers 70 years old or older who were recruited from clinic and community sources. Treatment assignment was concealed until eligibility was established. Participants randomized to intervention underwent two 4-hour classroom and two 1-hour on-road sessions focused on common problem areas of older drivers. Controls received modules directed at vehicle, home, and environmental safety. A knowledge test and driving performance were assessed at baseline and 8 weeks. On-road driving performance was assessed by an experienced evaluator in a dual-brake-equipped vehicle in urban, residential, and highway traffic. Driving performance was rated on a 36-item scale with potential scores from 0 to 72 (higher score better). The knowledge test included 20 road knowledge and eight road sign questions, scored from 0 to 28 correct. RESULTS: The least squares mean change in road test score relative to baseline was 2.87 points higher in the intervention than in the control group (p =.001). The least squares mean change in knowledge test scores relative to baseline was 3.45 points higher in the intervention than in the control group (p <.001). CONCLUSIONS: An education program consisting of classroom and on-road training targeted to common errors of older drivers enhanced performance on knowledge and on-road tests. Such interventions offer older drivers the potential to continue driving safely longer and to maintain their out-of-home mobility. PMID- 17921425 TI - Low plasma N-3 fatty acids and dementia in older persons: the InCHIANTI study. AB - BACKGROUND: N-3 fatty acids (FA) have an important role in brain development and function. However, there is conflicting evidence concerning the relationship between n-3 FA and dementia in older persons. METHODS: In the Invecchiare in Chianti (InCHIANTI) study, we measured plasma FA by gas chromatography in 935 community-dwelling older persons randomly extracted from the population of two towns near Florence, Italy. Cognitive impairment was measured using the Mini Mental Status Examination. Participants who scored /= 65 years old admitted to one Canadian health region hospital between July 1999 and September 2000. Participants completed the Modified Barthel Functional Index (MBI) in hospital and again via telephone interviews 6 months postoperatively. Data were also collected on surgery and rehabilitation timing, length of hospital stay (LOS). and discharge destination. Relative change from prefracture function adjusting for known prognostic factors, and the proportion of participants returning to prefracture function were compared between the LTC and community-dwelling residents. RESULTS: LTC residents (n = 115) were older, with lower function prefracture, more comorbidities, and increased dementia than community-dwelling residents (n = 336). Six months postfracture, 17 (22%) LTC and 180 (71%) community-dwelling residents had regained prefracture function (p <.001). LTC residents had 33% lower (-40.6, -27.2) and community-dwelling residents 11.6% lower (-14.8, -8.4) 6-month MBI scores relative to prefracture scores after risk adjustment. Residential status was significantly associated with risk-adjusted functional recovery (p <.001). Median LOS was 4 days less for LTC than for community-dwelling residents (p <.001). Twelve (10%) LTC and 266 (79%) community dwelling residents were discharged to inpatient rehabilitation (p <.001). CONCLUSION: Following hip fracture, most LTC residents do not regain prefracture function irrespective of known prognostic factors. Further investigation is needed as to the extent to which personal and environmental characteristics contribute to outcome after hip fracture. PMID- 17921427 TI - Executive function, more than global cognition, predicts functional decline and mortality in elderly women. AB - BACKGROUND: Functional impairment in community-dwelling older adults is common and is associated with poor outcomes. Our goal was to compare the contribution of impairment in executive function or global cognitive function to predicting functional decline and mortality. METHODS: We studied 7717 elderly women enrolled in a prospective study (mean age 73.3 years) and identified women with poor baseline executive function (score > 1 standard deviation [SD] below the mean on the Trail Making Test B (Trails B; n = 957, 12.4%), poor global cognitive function (score > 1 SD below the mean on a modified Mini-Mental State Examination [mMMSE], n = 387, 5.0%), impairment in both (n = 249, 3.2%), or no impairment (n = 6124, 79.4%). We compared level of functional difficulty (Activities of Daily Living [ADLs] and Instrumental ADLs [IADLs]) at baseline and at 6-year follow-up and survival at follow-up. We also determined if the association was independent of age, education, depression, medical comorbidities, and baseline functional ability. RESULTS: At baseline, women with Trails B impairment only or impairment on both tests reported the highest proportion of ADL and IADL dependence compared to the other groups. At the 6-year follow-up after adjusting for age, education, medical comorbidities, depression, and baseline ADL or IADL, women with only Trails B impairment were 1.3 times more likely to develop an incident ADL dependence (adjusted odds ratio [OR] = 1.34; 95% confidence interval [CI], 1.07 1.69) and 1.5 times more likely to develop a worsening of ADL dependence (adjusted OR = 1.48; 95% CI, 1.16-1.89) when compared to women with no impairment on either test. In addition, women with only Trails B impairment had a 1.5-fold increased risk of mortality (adjusted hazard ratio [HR] = 1.48; 95% CI, 1.21 1.81). In contrast, women with impairment on only mMMSE were not at increased risk to develop incident ADL or IADL dependence, a worsening of ADL or IADL dependence, or mortality. CONCLUSION: Compared to women with no impairment, women with executive function impairment had significantly worse ADL and IADL function cross-sectionally and over 6 years. Individuals with executive dysfunction also had increased risk of mortality. These results suggest that screening of executive function can help to identify women who are at risk for functional decline and decreased survival. PMID- 17921428 TI - Self-efficacy mediates walking performance in older adults with knee osteoarthritis. AB - BACKGROUND: Self-efficacy is a determinant of walking performance in older adults with knee osteoarthritis. We examined whether self-efficacy mediated the effect of age, psychosocial, impairment, and mechanical factors on walking performance. METHODS: Fifty-four participants with knee osteoarthritis completed the Six Minute Walk test and Arthritis Self-Efficacy Scale. Independent variables reflected age, psychosocial (depressive symptoms), impairment (pain, stiffness), and mechanical (strength, obesity) factors. RESULTS: Self-efficacy fully mediated the effect of age and impairments on walking. The effects of strength were only partially mediated by self-efficacy. Depressive symptoms and obesity were not mediated by self-efficacy. CONCLUSIONS: These findings are consistent with Social Cognitive Theory, according to which age may alter outcome expectations, and impairments like pain and stiffness provide negative physiological feedback to influence performance. Mechanical factors like strength and obesity may better represent a person's capabilities and interact with other variables to influence physical performance in older adults with knee osteoarthritis. PMID- 17921430 TI - Risk factors for methicillin-resistant Staphylococcus aureus colonization in a geriatric rehabilitation hospital. AB - BACKGROUND: Knowledge of the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) colonization, and the frequency of transmission is vital for the implementation of MRSA infection control measures in hospitals. We assessed risk factors for and rates of colonization of MRSA upon admission to a geriatric rehabilitation hospital, and studied the contribution of the colonization on hospital mortality. METHODS: This was a prospective study conducted over a 6 month period. All patients were screened at admission, using nasal and throat swabs. Whenever necessary, blood, sputum, urine, or wound cultures were obtained. Data collected on admission included age, sex, functional status, reasons for admission to the rehabilitation unit, previous hospitalizations, known carriage or infection with MRSA in previous hospitalizations, underlying diseases, prescribed antibiotics in previous hospitalization, serum albumin, creatinine clearance, and patient management data. Risk factors for MRSA carriage on admission were estimated by using multivariate analysis. RESULTS: Of the 337 patients admitted during the study period, 24 (7.1%) had a MRSA isolate, and 87.4% of these were detected through screening specimens only. Of the 24 positive admissions with MRSA, 23 (95.8%) were newly identified MRSA carriers. In the multivariate analysis, the following variables were highly associated with MRSA carriage: presence of tracheotomy (p =.0001), hospitalization for deconditioning (p =.007), renal failure (p =.039), and quinolone use prior to hospitalization (p =.037). The morbidity associated with MRSA was very low, and length of stay was not influenced by carriage. CONCLUSION: The prevalence of MRSA carriage on admission to geriatric rehabilitation hospitals is high. Screening on admission is probably useful, as it detects almost all MRSA carriers. However, given the low morbidity associated with MRSA observed in this study, eradication of the MRSA carrier state is questionable. Further studies are needed to determine the usefulness and cost/benefit ratio of screening. PMID- 17921429 TI - Reporting peak expiratory flow in older persons. AB - BACKGROUND: Peak expiratory flow ("peak flow") predicts important outcomes in older persons. Nevertheless, its clinical application is uncertain because prior strategies for reporting peak flow may not be valid. We thus determined the frequency distribution of peak flow by the conventional strategy of percent predicted (%predicted) and by an alternative method termed standardized residual (SR) percentile, and evaluated how these two metrics relate to health status in older persons. METHODS: Participants included 754 community-dwelling persons aged >/= 70 years. Data included chronic conditions, frailty indicators, and peak flow. RESULTS: Mean age was 78.4 years, with 63.7% reporting a smoking history, 17.4% chronic lung disease, and 77.1% having one or more frailty indicators. Peak flow >/= 80 %predicted was recorded in 67.5% of participants, whereas peak flow >/= 80th SR percentile was only noted in 15.9%. A graded relationship was observed between peak flow and health status, but %predicted yielded health risk at peak flows currently considered normal (80-100 %predicted), whereas SR percentile conferred health risk only at severely reduced peak flows (< 50th SR percentile). CONCLUSIONS: Peak flow expressed as SR percentile attains a frequency distribution more consistent with the characteristics of our elderly cohort, and establishes health risk at more appropriate levels of reduced peak flow. These findings establish the need for longitudinal studies based on SR percentile to further evaluate the use of peak flow as a risk assessment tool in older persons and to determine if pulmonary function, in general, is better reported in older persons as SR percentile, rather than as %predicted. PMID- 17921431 TI - Association between functional polymorphisms of renin-angiotensin system, left ventricular mass, and geometry over 4 years in a healthy Chinese population aged 60 years and older. AB - BACKGROUND: Cross-sectional studies investigated the impact of renin-angiotensin system (RAS) gene polymorphism on left ventricular mass index (LVMI) with conflicting results. We conducted a longitudinal study to investigate the influence of the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) and angiotensinogen (AGT) M235T and angiotensin II type 1 receptor (AT1R) A1166C gene polymorphisms on the LVMI and geometry. METHODS: Of 1500 people screened, 110 nondiabetic normotensive elderly Chinese persons were recruited and received echocardiography at baseline and at the 2nd and 4th year follow-up. No participants had a history of organic heart disease or chronic medication. The gene polymorphisms were analyzed by using polymerase chain reaction. RESULTS: Participant age was 71.9 +/- 3.9 years (range 60-81 years). The prevalence of concentric remodeling, eccentric hypertrophy, and concentric hypertrophy was significantly increased as well as LVMI after 4 years (all p <.05). These changes and the magnitude of LVMI increase were significantly higher in participants carrying the ACE D allele than non-D-allele carriers (all p <.05). This association was still significant in multivariate analyses (p /=60 years. METHODS: We searched English articles in Medline (1996-2004) indexed under "falls" or "accidental falls" and "pharmaceutical preparations" or specific groups of drugs. We excluded studies not meeting the age criterion, not controlled with nonusers of target medicines or nonfallers, or with no clear definition of target medication. RESULTS: Twenty eight observational studies and one randomized controlled trial met the inclusion criteria. The number of participants ranged from 70 to 132,873. The outcome measure was a fall in 22 studies and a fracture in 7 studies. The main group of drugs associated with an increased risk of falling was psychotropics: benzodiazepines, antidepressants, and antipsychotics. Antiepileptics and drugs that lower blood pressure were weakly associated with falls. CONCLUSIONS: Central nervous system drugs, especially psychotropics, seem to be associated with an increased risk of falls. The quality of observational studies needs to be improved, for many appear to lack even a clear definition of a fall, target medicines, or prospective follow-up. Many drugs commonly used by older persons are not systematically studied as risk factors for falls. PMID- 17921435 TI - Remodeling of apolipoprotein E-containing spherical reconstituted high density lipoproteins by phospholipid transfer protein. AB - Phospholipid transfer protein (PLTP) transfers phospholipids between HDL and other lipoproteins in plasma. It also remodels spherical, apolipoprotein A-I (apoA-I)-containing HDL into large and small particles in a process involving the dissociation of lipid-free/lipid-poor apoA-I. ApoE is another apolipoprotein that is mostly associated with large, spherical HDL that do not contain apoA-I. Three isoforms of apoE have been identified in human plasma: apoE2, apoE3, and apoE4. This study investigates the remodeling of spherical apoE-containing HDL by PLTP and the ability of PLTP to transfer phospholipids between apoE-containing HDL and phospholipid vesicles. Spherical reconstituted high density lipoproteins (rHDL) containing apoA-I [(A-I)rHDL], apoE2 [(E2)rHDL], apoE3 [(E3)rHDL], or apoE4 [(E4)rHDL] as the sole apolipoprotein were prepared by incubating discoidal rHDL with low density lipoproteins and lecithin:cholesterol acyltransferase. PLTP remodeled the spherical, apoE-containing rHDL into large and small particles without the dissociation of apoE. The PLTP-mediated remodeling of apoE-containing rHDL was more extensive than that of (A-I)rHDL. PLTP transferred phospholipids from small unilamellar vesicles to apoE-containing rHDL in an isoform-dependent manner, but at a rate slower than that for spherical (A-I)rHDL. It is concluded that apoE enhances the capacity of PLTP to remodel HDL but reduces the ability of HDL to participate in PLTP-mediated phospholipid transfers. PMID- 17921436 TI - Sterol-dependent regulation of proprotein convertase subtilisin/kexin type 9 expression by sterol-regulatory element binding protein-2. AB - Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a member of the subtilases that promotes the internalization and degradation of LDL receptor in liver and thereby controls the level of LDL cholesterol in plasma. Here, we show that the expression of PCSK9 in HepG2 cells is completely dependent on the absence or presence of sterols. The minimal promoter region of the PCSK9 gene contains a sterol-regulatory element (SRE), which makes the transcription of PCSK9 dependent on sterols. Expression of nuclear forms of sterol-regulatory element binding protein-1 (SREBP-1) and SREBP-2 dramatically increased the promoter activity of PCSK9. In vitro-translated nuclear forms of SREBPs showed interactions with SRE, whereas mutations in SRE abolished their binding. In vivo studies in mice showed that Pcsk9 protein and mRNA were decreased significantly by fasting and increased by refeeding. However, supplementation with 2% cholesterol in the diet prevented the increase in Pcsk9. The amounts of Pcsk9 mRNA in livers of refed mice showed correlated regulation by the changes in the nuclear form of Srebp-2. In summary, it is suggested that the expression of PCSK9 is regulated by sterol at the transcriptional level in HepG2 cells and that both SREBP-1 and SREBP-2 can transcriptionally activate PCSK9 via SRE in its proximal promoter region in vitro. However, in vivo, it is suggested that the sterol dependent regulation of PCSK9 is mediated predominantly by SREBP-2. PMID- 17921437 TI - Mammary glands of adipophilin-null mice produce an amino-terminally truncated form of adipophilin that mediates milk lipid droplet formation and secretion. AB - Adipophilin (ADPH), a member of the perilipin family of lipid droplet-associated proteins, is hypothesized to mediate milk lipid formation and secretion. Unexpectedly, the fat content of milk from ADPH-null mice was only modestly lower than that of wild-type controls, and neither TIP47 nor perilipin appeared to fully compensate for ADPH loss. This prompted us to investigate the possibility that the mutated ADPH gene was not a genuine null mutation. ADPH transcripts were detected in ADPH-null mammary tissue by quantitative real-time PCR, and C terminal-specific, but not N-terminal-specific, ADPH antibodies detected a single lower molecular weight product and immunostained cytoplasmic lipid droplets (CLDs) and secreted milk fat globules in ADPH-null mammary tissue. Furthermore, stable cell lines expressing cDNA constructs corresponding to the ADPH-null mutation produced a product comparable in size to the one detected in ADPH-null mammary glands and localized to CLDs. Based on these data, we conclude that ADPH null mice express an N-terminally truncated form of ADPH that retains the ability to promote the formation and secretion of milk lipids. PMID- 17921438 TI - Survey of the prevalence of Salmonella species on commercial laying farms in the United Kingdom. AB - A survey of salmonella infection on 454 commercial layer flock holdings in the uk was carried out between October 2004 and September 2005. Fifty-four (11.7 per cent, 95 per cent confidence interval 9.3 to 14.0 per cent) were salmonella positive. The most common serovar identified was Salmonella Enteritidis at a prevalence of 5.8 per cent, and 70 per cent of these isolates were phage types 4, 6, 7 and 35. Salmonella Typhimurium was the second most prevalent serovar, found in 1.8 per cent of the farms. Of the three other serovars given top priority by the eu because of their public health significance, Salmonella Virchow and Salmonella Infantis were each isolated from one holding, but Salmonella Hadar was not isolated from any of the holdings. Analysis of antimicrobial resistance patterns revealed that over 76 per cent of the isolates were sensitive to all of the 16 drugs tested, and all the isolates were sensitive to ciprofloxacin, gentamicin, ceftazidime, apramycin, amikacin, amoxicillin/clavulanic acid, neomycin and cefotaxime. PMID- 17921439 TI - Survey of the seroprevalence of brucellosis in ruminants in Tajikistan. AB - A cross-sectional serological survey of the prevalence of brucellosis in ruminants in the Region of Republican Subordination and Khatlon oblasts (provinces) in Tajikistan was conducted in May 2003. Sera from 13,625 ruminants involving 3513 households in 172 kishlaks (villages) were collected and screened by the rose bengal test. Doubtful and positive results were further tested with competitive and indirect elisas. The overall serological prevalences (95 per cent confidence intervals [cis]) were 5.8 per cent (5.2 to 6.4 per cent) for sheep, 5.5 per cent (5.0 to 6.0 per cent) for goats and 2.1 per cent (1.0 to 3.2 per cent) for cattle. The results show that brucellosis was a common disease of ruminants that was widely but unevenly distributed throughout the two oblasts. Seropositive animals were found in 119 of the 172 kishlaks (69.2 per cent [95 per cent ci 61.9 to 75.6 per cent]) and 14.4 per cent (95 per cent ci 13.3 to 15.6 per cent) of the 3513 households. Evidence of infection was also found in cattle kept for milk production in urban kishlaks in two major cities and in state-owned dairy farms. PMID- 17921440 TI - Spirorchiidiosis (Digenea: Spirorchiidae) and lesions associated with parasites in Caribbean green turtles (Chelonia mydas). AB - Forty-seven nesting green turtles (Chelonia mydas) were examined for parasitic pathogens. Four species of cardiovascular flukes (Digenea: Spirorchiidae), Learedius learedi, Hapalotrema postorchis, Monticellius indicum and Amphiorchis solus were collected from 39 of 40 of the turtles, and Neospirorchis species were identified in seven of the 47 by histological examination. The pathological changes associated with the spirorchiids and their eggs included aneurysms, arteritis of great vessels with dark nodular thickenings, endocarditis, haemorrhagic lesions, thrombi, vasculitis, and granulomatous reactions in all the turtles. Ozobranchus branchiatus (Hirudinea: Ozobranchidae) leeches were found on the skin of 27 of the 47 turtles and were associated with traumatic purulent and ulcerative dermatitis. Oesophagitis associated with Rameshwarotrema uterocrescens (Digenea: Pronocephalidae) was recorded in 11 of 43 of the turtles. Petechial haemorrhages and unspecific crater-like ulcerated lesions on the gastric mucosa associated with Charaxicephaloides species and Charaxicephalus robustus (Digenea: Pronocephalidae) were observed in four of 40 of the turtles. Cholycystitis and ectasia of mucosal glands associated with Rhytidodoides similis (Digenea: Rhytidodidae) were observed in one of the 47 turtles. Fibropapillomas were observed on the skin of one turtle and fibromas on the skin of two of them. PMID- 17921441 TI - Re-emergence of bluetongue in northern Europe in 2007. PMID- 17921442 TI - Prevalence of antibodies to Bartonella henselae in dogs in Italy. PMID- 17921443 TI - Effect of intravenous calcium and oral sodium phosphate on electrolytes in cows with parturient paresis. PMID- 17921444 TI - Malignant catarrhal fever in cattle in the UK. PMID- 17921445 TI - TB policy and the ISG's findings. PMID- 17921446 TI - Postweaning multisystemic wasting syndrome in Chile. PMID- 17921447 TI - Staphylococcus schleiferi subspecies coagulans in dogs. PMID- 17921448 TI - Opposite regulation of endothelial NO synthase by HSP90 and caveolin in liver and lungs of rats with hepatopulmonary syndrome. AB - The hepatopulmonary syndrome is a complication of cirrhosis that associates an overproduction of nitric oxide (NO) in lungs and a NO defect in the liver. Because endothelial NO synthase (eNOS) is regulated by caveolin that decreases and heat shock protein 90 (HSP90) that increases NO production, we hypothesized that an opposite regulation of eNOS by caveolin and HSP90 might explain the opposite NO production in both organs. Cirrhosis was induced by a chronic bile duct ligation (CBDL) performed 15, 30, and 60 days before sample collection and pharmacological tests. eNOS, caveolin, and HSP90 expression were measured in hepatic and lung tissues. Pharmacological tests to assess NO released by shear stress and by acetylcholine were performed in livers (n = 28) and lungs (n = 28) isolated from normal and CBDL rats. In lungs from CBDL rats, indirect evidence of high NO production induced by shear stress was associated with a high binding of HSP90 and a low binding of caveolin to eNOS. Opposite results were observed in livers from CBDL rats. Our study shows an opposite posttranslational regulation of eNOS by HSP90 and caveolin in lungs and liver from rats with CBDL. Such opposite posttranslational regulation of eNOS by regulatory proteins may explain in part the pulmonary overproduction of NO and the hepatic NO defect in rats with hepatopulmonary syndrome. PMID- 17921449 TI - Shared decision making, decision aids, and risk communication. PMID- 17921450 TI - Incorporating decision analysis in decision aids. PMID- 17921451 TI - Aiding medical decision making: a communication perspective. PMID- 17921452 TI - Contingent or universal approaches to patient deficiencies in health numeracy. PMID- 17921453 TI - Removal of FKBP12.6 does not alter the conductance and activation of the cardiac ryanodine receptor or the susceptibility to stress-induced ventricular arrhythmias. AB - The 12.6-kDa FK506-binding protein (FKBP12.6) is considered to be a key regulator of the cardiac ryanodine receptor (RyR2), but its precise role in RyR2 function is complex and controversial. In the present study we investigated the impact of FKBP12.6 removal on the properties of the RyR2 channel and the propensity for spontaneous Ca(2+) release and the occurrence of ventricular arrhythmias. Single channel recordings in lipid bilayers showed that FK506 treatment of recombinant RyR2 co-expressed with or without FKBP12.6 or native canine RyR2 did not induce long-lived subconductance states. [(3)H]Ryanodine binding studies revealed that coexpression with or without FKBP12.6 or treatment with or without FK506 did not alter the sensitivity of RyR2 to activation by Ca(2+) or caffeine. Furthermore, single cell Ca(2+) imaging analyses demonstrated that HEK293 cells co-expressing RyR2 and FKBP12.6 or expressing RyR2 alone displayed the same propensity for spontaneous Ca(2+) release or store overload-induced Ca(2+) release (SOICR). FK506 increased the amplitude and decreased the frequency of SOICR in HEK293 cells expressing RyR2 with or without FKBP12.6, indicating that the action of FK506 on SOICR is independent of FKBP12.6. As with recombinant RyR2, the conductance and ligand-gating properties of single RyR2 channels from FKBP12.6 null mice were indistinguishable from those of single wild type channels. Moreover, FKBP12.6-null mice did not exhibit enhanced susceptibility to stress induced ventricular arrhythmias, in contrast to previous reports. Collectively, our results demonstrate that the loss of FKBP12.6 has no significant effect on the conduction and activation of RyR2 or the propensity for spontaneous Ca(2+) release and stress-induced ventricular arrhythmias. PMID- 17921454 TI - Spatiotemporal expression of ameloblastin isoforms during murine tooth development. AB - Ameloblasts synthesize and secrete the enamel matrix proteins (amelogenin, ameloblastin, and enamelin). This investigation examined the profiles of ameloblastin in the ameloblasts and in the enamel matrix during different postnatal (PN) days (days 0-9) of development of mouse molar, using an antibody specific for C-terminal sequence of ameloblastin (Ct; GNKVHQPQVHNAWRF). Ameloblastin is found in three different molecular sizes (37, 55, and 66 kDa) in both ameloblasts and enamel matrix during PN development. In the ameloblasts, the sequence of expression of these fractions varied. The 37-kDa fraction was observed (even before the appearances of mRNA of the proteases, enamelysin and kallikrein-4) on days 0 and 1, persisted until day 3, and was not found thereafter. Other isoforms (55 and 66 kDa) distinctly appeared in ameloblasts after day 1, reached a peak on day 5, and remained thereafter. The Ct-positive granules appeared beaded in the ameloblasts on day 3. In the extracellular matrix, a 37-kDa (but not 66- or 55-kDa) fraction was detected on days 0 and 1 and remained in the matrix throughout the PN days. The larger isoforms (55 and 66 kDa) appeared in the enamel matrix from day 3 onward. On days 0-3, but not later, the 37-kDa isoform co-localizes with amelogenin in Tomes' process and formative enamel, as revealed by laser scan confocal microscopy. Autoradiography confirmed accumulation of 3H-labeled amelogenin trityrosyl motif peptide in the region of Tomes' process and formative enamel from day 0 to 3. These observations suggest that the 37-kDa isoform interacts with amelogenin during early tooth development. PMID- 17921455 TI - Deep sulcal landmarks provide an organizing framework for human cortical folding. AB - The folding pattern of the cerebral cortex and its relation to functional areas is notoriously variable and there is a need to identify more consistent 3 dimensional (3D) topographical cortical features. We analyzed magnetic resonance brain images of 96 normal adult human volunteers using automated 3D image analysis methods. We examined the deeper parts of the sulci because they generally show less interindividual variability than more superficial parts, especially in monozygotic twins, and deepest parts of primary sulci are the first to develop embryologically and change least as the cortex expands. Along the length of each sulcus we found that there is generally one well-defined zone where depth is maximal, which we term the sulcal pit. Long sulci may have 2 or 3 pits. The spatial arrangement of pits is strikingly regular, forming alternating chains of deeper and shallower pits. We hypothesize that the pits are encoded in the protomap described in Rakic (1988. Specification of cerebral cortical areas. Science. 241:170-176) and are under closer genetic control than the rest of the cortex and are likely to have a more consistent relationship to functional areas. PMID- 17921457 TI - A discrete area within the left dorsolateral prefrontal cortex involved in visual verbal incongruence judgment. AB - The role of the frontal lobe in cross-modal visual-auditory processing has been documented in experiments using incongruent/congruent paradigms. In this study, 4 patients with left frontal World Health Organization Grade II glioma were assessed during pre-, intra-, and postoperative sessions with picture-naming and verbal-visual task requiring judgment of congruence between pictures and words. During awake brain surgery, the naming and cross-modal tasks were coupled with electrical stimulation inactivating restricted specific regions. For all patients, focal brain stimulation in the dorsolateral prefrontal cortex elicited picture-word matching disturbances but no naming impairment, and the elicited errors exclusively appeared in incongruent and not congruent conditions. The dissociation observed between correct picture naming and disturbed cross-modal judgment shows that electrical stimulation of a discrete cortical area within the left dorsolateral prefrontal cortex can inhibit the simultaneous processing of visual-verbal information without disturbing larger networks involved in the naming process. PMID- 17921458 TI - Plasticity of representational maps in somatosensory cortex observed by in vivo voltage-sensitive dye imaging. AB - We investigated the effect of selective whisker trimming on the development of the cortical representation of a whisker deflection in layer 2/3 of rat somatosensory cortex using in vivo voltage-sensitive dye (vsd) imaging. Responses to deflection of D-row whiskers were recorded after trimming of A-row, B-row, and C-row whiskers, referred to as DE pairing, during postnatal development. Animals DE paired from postnatal day (p) 7 to p17 had a significant bias in the spread of the vsd signal, favoring spread toward the concomitantly nondeprived E-row columns. This resulted primarily from a strong decrease in signal spreading into the deprived C-row columns. In contrast, signal spread in control littermates was approximately symmetrical. DE pairing failed to elicit significant changes when begun after p14, thus defining a critical period for this phenomenon. The results suggest that sensory deprivation in this model results in lower connectivity being established between nondeprived columns and adjacent deprived ones. PMID- 17921460 TI - Hydration and health promotion. Proceedings of the International Life Sciences Institute North America Conference on Hydration and Health Promotion. November 29 30, 2006. Washington, DC, USA. PMID- 17921459 TI - Role in anxiety behavior of the endocannabinoid system in the prefrontal cortex. AB - In the present study we explored with a multidisciplinary approach, the role of anandamide (AEA) in the modulation of anxiety behavior at the level of the prefrontal cortex (PFC). Low doses of the metabolically stable AEA analog, methanandamide, microinjected into the PFC, produced an anxiolytic-like response in rats, whereas higher doses induced anxiety-like behaviors. Pretreatment with the selective antagonist of CB1 or TRPV1 receptors (AM251 and capsazepine, respectively) suggested that the anxiolytic effect evoked by AEA might be due to the interaction with the CB1 cannabinoid receptor, whereas vanilloid receptors seem to be involved in AEA anxiogenic action. When AEA contents in the PFC were increased by microinjecting the selective inhibitor of fatty acid amide hydrolase (FAAH), URB597, we observed an anxiolytic response only at low doses of the compound and no effect or even an anxiogenic profile at higher doses. In line with this, a marked decrease of AEA levels in the PFC, achieved by lentivirus mediated local overexpression of FAAH, produced an anxiogenic response. These findings support an anxiolytic role for physiological increases in AEA in the PFC, whereas more marked increases or decreases of this endocannabinoid might lead to an anxiogenic response due to TRPV1 stimulation or the lack of CB1 activation, respectively. PMID- 17921456 TI - Cortical activity time locked to the shift and maintenance of spatial attention. AB - Attention increases the gain of visual neurons, which improves visual performance. How attention is controlled, however, remains unknown. Clear correlations between attention and saccade planning indicate that the control of attention is mediated through mechanisms housed in the oculomotor network. Here, we used event-related functional magnetic resonance imaging to compare overt and covert attention shifts. Subjects covertly or overtly shifted attention based on an endogenous cue and maintained attention throughout a long and variable delay. To insure continued attention, subjects counted when the attended target dimmed at near-threshold contrast levels. Overt and covert tasks used identical stimuli and required identical motor responses. Additionally, a staircase procedure that adjusted the target-dimming contrast separately for covert and overt trials equated the difficulty between conditions and across subjects. We found that the same regions along the precentral and intraparietal sulci were active during shifts of covert and overt attention. We also found sustained activation in the hemisphere contralateral to the attended visual field. We conclude that maps of prioritized locations are represented in areas classically associated with oculomotor control. The readout of these spatial maps by posterior visual areas directs spatial attention just as the readout by downstream saccade generators directs saccades. PMID- 17921461 TI - Hydration and health promotion: a brief introduction. PMID- 17921462 TI - Hydration and disease. AB - Many diseases have multifactorial origins. There is increasing evidence that mild dehydration plays a role in the development of various morbidities. In this review, effects of hydration status on acute and chronic diseases are depicted (excluding the acute effects of mild dehydration on exercise performance, wellness, cognitive function, and mental performance) and categorized according to four categories of evidence (I-IV). Avoidance of a high fluid intake as a precautionary measure may be indicated in patients with cardiovascular disorders, pronounced chronic renal failure (III), hypoalbuminemia, endocrinopathies, or in tumor patients with cisplatin therapy (IIb) and menace of water intoxication. Acute systemic mild hypohydration or dehydration may be a pathogenic factor in oligohydramnios (IIa), prolonged labor (IIa), cystic fibrosis (III), hypertonic dehydration (III), and renal toxicity of xenobiotica (Ib). Maintaining good hydration status has been shown to positively affect urolithiasis (Ib) and may be beneficial in treating urinary tract infection (IIb), constipation (III), hypertension (III), venous thromboembolism (III), fatal coronary heart disease (III), stroke (III), dental disease (IV), hyperosmolar hyperglycemic diabetic ketoacidosis (IIb), gallstone disease (III), mitral valve prolapse (IIb), and glaucoma (III). Local mild hypohydration or dehydration may play a critical role in the pathogenesis of several broncho-pulmonary disorders like exercise asthma (IIb) or cystic fibrosis (Ib). In bladder and colon cancers, the evidence on hydration status' effects is inconsistent. PMID- 17921463 TI - Hydration and physical performance. AB - There is a rich scientific literature regarding hydration status and physical function that began in the late 1800s, although the relationship was likely apparent centuries before that. A decrease in body water from normal levels (often referred to as dehydration or hypohydration) provokes changes in cardiovascular, thermoregulatory, metabolic, and central nervous function that become increasingly greater as dehydration worsens. Similarly, performance impairment often reported with modest dehydration (e.g., -2% body mass) is also exacerbated by greater fluid loss. Dehydration during physical activity in the heat provokes greater performance decrements than similar activity in cooler conditions, a difference thought to be due, at least in part, to greater cardiovascular and thermoregulatory strain associated with heat exposure. There is little doubt that performance during prolonged, continuous exercise in the heat is impaired by levels of dehydration >or= -2% body mass, and there is some evidence that lower levels of dehydration can also impair performance even during relatively short-duration, intermittent exercise. Although additional research is needed to more fully understand low-level dehydration's effects on physical performance, one can generalize that when performance is at stake, it is better to be well-hydrated than dehydrated. This generalization holds true in the occupational, military, and sports settings. PMID- 17921464 TI - Dehydration and cognitive performance. AB - Human neuropsychology investigates brain-behavior relationships, using objective tools (neurological tests) to tie the biological and behavior aspects together. The use of neuropsychological assessment tools in assessing potential effects of dehydration is a natural progression of the scientific pursuit to understand the physical and mental ramifications of dehydration. It has long been known that dehydration negatively affects physical performance. Examining the effects of hydration status on cognitive function is a relatively new area of research, resulting in part from our increased understanding of hydration's impact on physical performance and advances in the discipline of cognitive neuropsychology. The available research in this area, albeit sparse, indicates that decrements in physical, visuomotor, psychomotor, and cognitive performance can occur when 2% or more of body weight is lost due to water restriction, heat, and/or physical exertion. Additional research is needed, especially studies designed to reduce, if not remove, the limitations of studies conducted to date. PMID- 17921465 TI - Hydration and cognition: a critical review and recommendations for future research. AB - The limited literature on the effects of dehydration on human cognitive function is contradictory and inconsistent. Although it has been suggested that decrements in cognitive performance are present in the range of a 2 to 3% reduction in body weight, several dose-response studies indicate dehydration levels of 1% may adversely affect cognitive performance. When a 2% or more reduction in body weight is induced by heat and exercise exposure, decrements in visual-motor tracking, short-term memory and attention are reported, but not all studies find behavioral effects in this range. Future research should be conducted using dose response designs and state-of-the-art behavioral methods to determine the lowest levels of dehydration that produce substantive effects on cognitive performance and mood. Confounding factors, such as caffeine intake and the methods used to produce dehydration, need to be considered in the design and conduct of such studies. Inclusion of a positive control condition, such as alcohol intake, a hypnotic drug, or other treatments known to produce adverse changes in cognitive performance should be included in such studies. To the extent possible, efforts to blind both volunteers and investigators should be an important consideration in study design. PMID- 17921466 TI - Hydration in children. AB - Water supply is a basic public problem. In modern science, three periods with different approaches to define recommended water intake in adults can be distinguished. Pediatricians agree that hydration in children may be optimal only in breastfed infants. More data are required on the health effects of different hydration states and varying water intakes in particular age and gender groups to define optimal ranges of water intake. The fetus grows in an exceptionally well hydrated environment. Water metabolism shows several peculiarities in preterm and term infants. Infant diarrhea remains a major topic of basic and clinical research. Water intoxication in infants, toddlers, and children is rare and can only be found in exceptional circumstances. Hydration status characterized by hyponatremia may play a role in the pathogenesis of febrile convulsions in toddlers. There is increasing indirect evidence that spontaneous drinking behavior of a population may be fixed and anchored in the age range of toddlers. Sex differences in hydration status are common, but not obligatory. What causes theses differences? What is behind the various circadian rhythms of urine osmolality in children? At what age and in what quantities can alcohol and caffeine consumption be tolerated? How can individual susceptibility be defined? Reflecting on the modern epidemic of obesity in children and adolescents, a public consensus concerning use and misuse of sweetened drinks seems mandatory. Dietary reference intakes of water refer to 24-hour intake. In nutritional counselling, food and meal-based dietary advice is primarily given. Young parents are confronted with a flood of advice of varying quality. Recommendations on fluid consumption should be collated and revised. PMID- 17921467 TI - How primary care providers might review evidence on hydration. AB - Primary care providers (PCPs) are increasing their use of evidence-based medicine (EBM) in the care they give patients. They evaluate the available evidence to determine if it applies to their patients and seek to complement their clinical experience with EBM to improve patient outcomes. In evidence-based practices, patient oriented data are valued more highly than disease oriented evidence. More than 8 million biomedical articles are published annually, but only an estimated 2% of those are relevant to improved patient outcomes (POEMs - patient oriented evidence that matters). This paper describes some of the tools used by PCPs to search for evidence and the decision-making process used to determine if they will change their practice. Understanding how PCPs evaluate research findings and other evidence can help hydration researchers frame their research questions and study reports. PMID- 17921468 TI - Assessing hydration status: the elusive gold standard. AB - Acknowledging that total body water (TBW) turnover is complex, and that no measurement is valid for all situations, this review evaluates 13 hydration assessment techniques. Although validated laboratory methods exist for TBW and extracellular volume, no evidence incontrovertibly demonstrates that any concentration measurement, including plasma osmolality (P(osm)), accurately represents TBW gain and loss during daily activities. Further, one blood or urine sample cannot validly represent fluctuating TBW and fluid compartments. Future research should (a) evaluate novel techniques that assess hydration in real time and are precise, accurate, reliable, non-invasive, portable, inexpensive, safe, and simple; and (b) clarify the relationship between P(osm) and TBW oscillations in various scenarios. PMID- 17921469 TI - Hydration needs throughout the lifespan. AB - In 2004, the Institute of Medicine (IOM) issued a report presenting reference intake information about water, sodium, potassium, and other electrolytes for Americans and Canadians. The report established, for the first time, adequate intakes (AI) for total water (from drinking water, beverages, and food) for healthy people. An AI is the recommended average daily intake level based on observed or experimentally-determined approximations or estimates of nutrient intake by a group or groups of apparently health people that are assumed to be adequate. The AI for water is based on US survey data. The recommendations about water intake are for healthy populations--they do not apply to at-risk groups. PMID- 17921470 TI - Limitations of data on fluid intake. AB - BACKGROUND: The Dietary Reference Intakes for Water, Potassium, Sodium, Chloride, and Sulfate, provided extensive tables of data on total water intake, water intake from beverages and water intake from foods; however, very little information was provided about the specific details of these analyses. Reported total water intake varied with gender and somewhat with age and suggested that further sources of variation in total water intake were due to differences in body size, physical activity, and climatic exposure. OBJECTIVE AND METHODS: In an attempt to better understand the variation in reported water intake, we examined water intake from foods and beverages and plain water intake in the National Health and Nutrition Examination Surveys (NHANES), 1999-2002. RESULTS: Non Hispanic black children (4 to 18 years) had less water from food and beverages (approximately 0.15 L/day) than non-Hispanic whites and Hispanics. Non-Hispanic black children had less (approximately 0.15 L/day) plain water than other ethnicities. Adult non-Hispanic whites had more water intake from food and beverages (approximately 0.4 L/day) and non-Hispanic blacks had less (approximately 0.2 L/day) than Hispanics and Other ethnic group. Non-Hispanic black adults had less (approximately 0.2 L/day) plain water than non-Hispanic whites and the Other ethnic group but had similar plain water consumption as Hispanics. CONCLUSION: These data indicate further investigation into the sources of variation of water intake should include consideration of ethnic differences. PMID- 17921471 TI - Role of whole foods in promoting hydration after exercise in humans. AB - Various reports indicate that humans receive 20-25% of their daily water intake from food. Fruits, vegetables and other high-moisture foods, therefore, make an important contribution to total fluid intake. In addition, co-ingestion of other nutrients and ingredients can impact drinking behavior, absorption, distribution and retention of water, all of which contribute to the person's hydration state. Therefore, a food's hydration value derives from the interaction between its water content and the presence of these co-nutrients and ingredients. Research is reviewed in this paper showing increased voluntary fluid intake of young boys during exercise when the beverage is flavored and contains sodium chloride and carbohydrate. Additional research on rehydration after exercise and heat exposure showed improved recovery of plasma volume and fluid status when food was ingested before consuming water in the two hours after exercise. Collectively, these findings point to an interaction between fluid intake and co-ingested nutrients in regulating human hydration during and after exercise. PMID- 17921472 TI - Hydration at the work site. AB - When performing physical work, sweat output often exceeds water intake, producing a body water deficit or dehydration. Specific to the work place, dehydration can adversely affect worker productivity, safety, and morale. Legislative bodies in North America such as the Occupational Safety and Health Administration (OSHA) and the American Conference of Governmental Industrial Hygienists (ACGIH) recommend replacing fluids frequently when exposed to heat stress, such as one cup (250 ml) every 20 minutes when working in warm environments. However, the majority of legislative guidelines provide vague guidance and none take into account the effects of work intensity, specific environments, or protective clothing. Improved occupational guidelines for fluid and electrolyte replacement during hot weather occupational activities should be developed to include recommendations for fluid consumption before, during, and after work. PMID- 17921473 TI - Exercise, heat, hydration and the brain. AB - The performance of both physical and mental tasks can be adversely affected by heat and by dehydration. There are well-recognized effects of heat and hydration status on the cardiovascular and thermoregulatory systems that can account for the decreased performance and increased sensation of effort that are experienced in the heat. Provision of fluids of appropriate composition in appropriate amounts can prevent dehydration and can greatly reduce the adverse effects of heat stress. There is growing evidence that the effects of high ambient temperature and dehydration on exercise performance may be mediated by effects on the central nervous system. This seems to involve serotonergic and dopaminergic functions. Recent evidence suggests that the integrity of the blood brain barrier may be compromised by combined heat stress and dehydration, and this may play a role in limiting performance in the heat. PMID- 17921474 TI - Mechanisms and significance of cell volume regulation. AB - Survival of human and animal cells requires avoidance of excessive alterations of cell volume. The osmolarity amassed by cellular accumulation of organic substances must be compensated by lowering cytosolic ion concentrations. The Na+/K+ ATPase extrudes Na+ in exchange for K+, which can permeate the cell membrane through K+ channels. K+ exit generates a cell-negative potential difference across the cell membrane, driving the exit of anions such as Cl-. The low cytosolic Cl- concentrations counterbalance the excess cellular osmolarity by organic substances. Cell volume regulation following cell swelling involves releasing ions through activation of K+ channels and/or anion channels, KCl cotransport, or parallel activation of K+/H+ exchange and Cl-/HCO3- exchange. Cell volume regulation following cell shrinkage involves accumulation of ions through activation of Na+,K+,2Cl- cotransport, Na+/H+ exchange in parallel to Cl /HCO3- exchange, or Na+ channels. The Na+ taken up is extruded by the Na+/K+ ATPase in exchange for K+. Shrunken cells further accumulate organic osmolytes such as sorbitol and glycerophosphorylcholine, and monomeric amino acids by altered metabolism and myoinositol (inositol), betaine, taurine, and amino acids by Na+ coupled transport. They release osmolytes during cell swelling. Challenges of cell volume homeostasis include transport, hormones, transmitters, and drugs. Moreover, alterations of cell volume participate in the machinery regulating cell proliferation and apoptotic cell death. Deranged cell volume regulation significantly contributes to the pathophysiology of several disorders such as liver insufficiency, diabetic ketoacidosis, hypercatabolism, fibrosing disease, sickle cell anemia, and infection. PMID- 17921475 TI - The molecular analysis of the shade avoidance syndrome in the grasses has begun. AB - The shade avoidance syndrome (SAS) is a morphological and physiological response initiated by a decrease in light quantity and a change in light quality. Recent work in Arabidopsis thaliana has begun to define the molecular components of the SAS in a model dicot species, but little is known of these networks in agronomically important grasses. The focus of this review is to present a current view of the SAS in the grasses based largely on the characterization of mutants in the phytochrome signal transduction pathway and on the effects of far-red light treatments on plant growth. In cereal grasses, intense selection by plant breeders has acted to attenuate some but not all shade avoidance responses within modern crop varieties. Traditionally, breeding efforts have been focused on optimizing grain yield. However, with the recent interest in lignocellulosic based biofuels, a new breeding paradigm may emerge to optimize biomass at the expense of grain yield. Some of the opportunities and challenges for engineering plant architecture to maximize resource use efficiency and yield by targeting the SAS in grasses are discussed. PMID- 17921476 TI - Proteomic analysis of tomato (Lycopersicon esculentum) pollen. AB - In flowering plants, pollen grains are produced in the anther and released to the external environment with the primary function of delivering sperm cells to the female gametophyte. This study was conducted to identify proteins in tomato pollen and to analyse their roles in relation to pollen function. Tomato is an important crop which is grown worldwide and is an excellent experimental system. Proteins were extracted from pollen, separated by two-dimensional gel electrophoresis (2-DE), and identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and peptide mass fingerprinting. Of the 960 spots observed on Colloidal Coomassie Blue (CCB) stained 2-DE gels, 190 were selected for analysis. Of these, 158 spots, representing 133 distinct proteins, were identified by searching the NCBInr and Expressed Sequence Tag databases. The identified proteins were classified based on designated functions and the majority included those involved in defence mechanisms, energy conversions, protein synthesis and processing, cytoskeleton formation, Ca(2+) signalling, and as allergens. A number of proteins in tomato pollen were similar to those reported in the pollen of other species; however, several additional proteins with roles in defence mechanisms, metabolic processes, and hormone signalling were identified. The potential roles of the identified proteins in the survival strategy of the small, independent, two celled pollen grain of tomato, and subsequently in pollen germination and tube growth are discussed. PMID- 17921477 TI - Risperidone versus olanzapine for treatment of schizophrenia. PMID- 17921478 TI - Cross-platform gene expression signature of human spermatogenic failure reveals inflammatory-like response. AB - BACKGROUND: The molecular basis of human testicular dysfunction is largely unknown. Global gene expression profiling of testicular biopsies might reveal an expression signature of spermatogenic failure in azoospermic men. METHODS: Sixty nine individual testicular biopsy samples were analysed on two microarray platforms; selected genes were validated by quantitative real-time PCR and immunohistochemistry. RESULTS: A minimum of 188 transcripts were significantly increased on both platforms. Their levels increased with the severity of spermatogenic damage and reached maximum levels in samples with Sertoli-cell-only appearance, pointing to genes expressed in somatic testicular cells. Over represented functional annotation terms were steroid metabolism, innate defence and immune response, focal adhesion, antigen processing and presentation and mitogen-activated protein kinase K signalling pathway. For a considerable proportion of genes included in the expression signature, individual transcript levels were in keeping with the individual mast cell numbers of the biopsies. When tested on three disparate microarray data sets, the gene expression signature was able to clearly distinguish normal from defective spermatogenesis. More than 90% of biopsy samples clustered correctly into the corresponding category, emphasizing the robustness of our data. CONCLUSIONS: A gene expression signature of human spermatogenic failure was revealed which comprised well studied examples of inflammation-related genes also increased in other pathologies, including autoimmune diseases. PMID- 17921479 TI - Estrogen metabolizing enzymes in endometrium and endometriosis. AB - BACKGROUND: Estradiol (E(2)) is an important promoter of the growth of both eutopic and ectopic endometrium. The findings with regard to the expression and activity of steroidogenic enzymes in endometrium of controls, in endometrium of endometriosis patients and in endometriotic lesions are not consistent. METHODS: In this study, we have looked at the mRNA expression and protein levels of a range of steroidogenic enzymes [aromatase, 17beta-hydroxysteroid dehydrogenases (17beta-HSD) type 1, 2 and 4, estrogen sulfotransferase (EST) and steroid sulfatase (STS)] in eutopic and ectopic endometrium of patients (n = 14) with deep-infiltrative endometriosis as well as in disease-free endometrium (n = 48) using real-time PCR and immunocytochemistry. In addition, we evaluated their menstrual cycle-related expression patterns, and investigated their steroid responsiveness in explant cultures. RESULTS: Aromatase and 17beta-HSD type 1 mRNA levels were extremely low in normal human endometrium, while mRNAs for types 2 and 4 17beta-HSD, EST and STS were readily detectable. Only 17beta-HSD type 2 and EST genes showed sensitivity to progesterone in normal endometrium. Types 1 and 2 17beta-HSD and STS protein was detected in normal endometrium using new polyclonal antibodies. CONCLUSIONS: In endometriosis lesions, the balance is tilted in favor of enzymes producing E(2). This is due to a suppression of types 2 and 4 17beta-HSD, and an increased expression of aromatase and type 1 17beta HSD in ectopic endometrium. PMID- 17921480 TI - Endometrial vessel maturation in women exposed to levonorgestrel-releasing intrauterine system for a short or prolonged period of time. AB - BACKGROUND: Levonorgestrel-releasing intrauterine system (LNG-IUS), although inserted to reduce heavy menstruation, causes irregular early transient bleeding. The objective of the study was to document quantitative changes in endometrial vessels of short- (< or =3 months) and long-term (> or =12 months) LNG users. The area, density and maturation of endometrial vessels were quantified in 19 endometrial biopsies of women with LNG-IUS and in 10 normally ovulating patients during mid-luteal phase. METHODS: Vessel maturation was evaluated by double immunostaining using anti-von Willebrand factor (endothelial cell marker) and anti-alpha Smooth Muscle Actin (vascular smooth muscle cells) antibodies. Vessel area, number and density were quantified with a novel computer-assisted image analysis system. RESULTS: Endometrium exposed to LNG-IUS for 1-3 months displayed a 11.5-fold increase in small naked vessel number. The partially mature vessel (alphaSMA partially positive) number increased six times. After long-term LNG-IUS treatment, the immature and partially mature vessel number remained four times higher than in the control group. Vessel area and density also increased dramatically in a time-dependent pattern with LNG-IUS use. CONCLUSIONS: Levonorgestrel affects blood vessel number, area, density and maturation in a time-dependent pattern that may explain the early transient increase in breakthrough bleeding with the LNG-IUS. PMID- 17921481 TI - Endometrial cells from women with endometriosis have increased adhesion and proliferative capacity in response to extracellular matrix components: towards a mechanistic model for endometriosis progression. AB - BACKGROUND: Endometriosis, classified as the presence of endometrial cells in ectopic sites, is a debilitating disease causing pain and infertility in approximately 10% of women of reproductive age. It is associated with the aberrant expression of extracellular matrix (ECM) components and their receptors, integrins. METHODS: We analysed the expression of integrins in stromal cells derived from peritoneal, ovarian and deeply infiltrating endometriotic lesions and from endometrium from women with and without endometriosis in vitro, using quantitative immunocytochemistry. The adhesive and proliferative capacity of each of the cell types in response to ECM components was assessed by in vitro assays of cell attachment and DNA synthesis. RESULTS: We demonstrate that eutopic and ectopic endometrial stromal cells from women with endometriosis exhibit an aberrant integrin profile in vitro compared with stromal cells derived from healthy controls. In addition, the former display increased adhesion and proliferative capacity in response to specific ECM components. CONCLUSIONS: We propose that the increased adhesive and proliferative potential of cells from endometriotic lesions may be a key feature in the pathogenesis of endometriosis. Furthermore, the elevated responsiveness of eutopic cells from women with endometriosis may contribute to the predisposition of some women to the disease. PMID- 17921482 TI - Analysis of rare amino acid replacements supports the Coelomata clade. AB - The recent analysis of a novel class of rare genomic changes, RGC_CAMs (after conserved amino acids-multiple substitutions), supported the Coelomata clade of animals as opposed to the Ecdysozoa clade (Rogozin et al. 2007). A subsequent reanalysis, with the sequences from the sea anemone Nematostella vectensis included in the set of outgroup species, suggested that this result was an artifact caused by reverse amino replacements and claimed support for Ecdysozoa (Irimia et al. 2007). We show that the internal branch connecting the sea anemone to the bilaterian animals is extremely short, resulting in a weak statistical support for the Coelomata clade. Direct estimation of the level of homoplasy, combined with taxon sampling with different sets of outgroup species, reinforces the support for Coelomata, whereas the effect of reversals is shown to be relatively minor. PMID- 17921484 TI - Relaxed constraint and evolutionary rate variation between basic helix-loop-helix floral anthocyanin regulators in Ipomoea. AB - Regulatory genes are believed to play a large role in morphological diversification and are often characterized by elevated rates of evolution. Whether this rapid evolution is primarily due to adaptive differentiation or relaxed selective constraint remains an open question. We attempted to distinguish between these alternative outcomes in 2 transcription factors known to regulate the expression of anthocyanin pigmentation genes in flowers. We cloned the full-length coding region from 2 basic helix-loop-helix transcription factors from several species of Ipomoea with diverse flower colors and determined the selective forces operating on them. In both genes, rapidly evolving sites and indel mutations are clustered in nonbinding domains, but the extent of rate acceleration in these domains is reduced relative to most previously characterized plant transcription factors. Moreover, codon models of substitution rates and models evaluating the magnitude of change to physical amino acid properties demonstrate little evidence for adaptive evolution and suggest that elevated nonsynonymous substitution rates in these domains represent relaxed selective constraint. Although both genes show qualitatively similar patterns, their rates of evolution differ significantly due to an increased rate of nonsynonymous substitutions in the nonbinding domains in one copy, suggesting substantial differences in functional constraint on each gene. In general, these results provide additional evidence demonstrating that decreased constraint as opposed to positive selection is largely responsible for the frequently observed pattern of rapid evolution in particular domains of plant transcription factors. More specifically, they suggest that most of the amino acid substitutions are neutral and do not implicate a role for natural selection on these regulatory genes in the diversification of flower color in Ipomoea. PMID- 17921483 TI - Protist homologs of the meiotic Spo11 gene and topoisomerase VI reveal an evolutionary history of gene duplication and lineage-specific loss. AB - Spo11 is a meiotic protein of fundamental importance as it is a conserved meiosis specific transesterase required for meiotic recombination initiation in fungi, animals, and plants. Spo11 is homologous to the archaebacterial topoisomerase VIA (Top6A) gene, and its homologs are broadly distributed among eukaryotes, with some eukaryotes having more than one homolog. However, the evolutionary relationships among these genes are unclear, with some debate as to whether eukaryotic homologs originated by lateral gene transfer. We have identified and characterized protist Spo11 homologs by degenerate polymerase chain reaction (PCR) and sequencing and by analyses of sequences from public databases. Our phylogenetic analyses show that Spo11 homologs evolved by two ancient eukaryotic gene duplication events prior to the last common ancestor of extant eukaryotes, resulting in three eukaryotic paralogs: Spo11-1, Spo11-2, and Spo11-3. Spo11-1 orthologs encode meiosis-specific proteins and are distributed broadly among eukaryotic lineages, though Spo11-1 is absent from some protists. This absence coincides with the presence of Spo11-2 orthologs, which are meiosis-specific in Arabidopsis and are found in plants, red algae, and some protists but absent in animals and fungi. Spo11-3 encodes a Top6A subunit that interacts with topoisomerase VIB (Top6B) subunits, which together play a role in vegetative growth in Arabidopsis. We identified Spo11-3 (Top6A) and Top6B homologs in plants, red algae, and a few protists, establishing a broader distribution of these genes among eukaryotes, indicating their likely vertical descent followed by lineage-specific loss. PMID- 17921485 TI - The repertoire of minimal mobile elements in the Neisseria species and evidence that these are involved in horizontal gene transfer in other bacteria. AB - In the Neisseria spp., natural competence for transformation and homologous recombination generate antigenic variants through creation of mosaic genes (such as opas) and through recombination with silent cassettes (such as pilE/pilS) and gene-complement diversity through the horizontal exchange of whole genes or groups of genes, in minimal mobile elements (MMEs). An MME is a region encompassing 2 conserved genes between which different whole-gene cassettes are found in different strains, which are chromosomally incorporated solely through the action of homologous recombination. Comparative analyses of the neisserial genome sequences identified 39 potential MME sites, the contents of which were investigated in 11 neisserial strains. One hundred and eight different MME regions were identified, 20 of which contain novel sequences and these contain 12 newly identified neisserial coding sequences. Neisserial uptake signal sequences are associated with 38 of the 40 MMEs studied. In some sites, divergent dinucleotide signatures of the sequences between the flanking genes suggest relatively recent horizontal acquisition of some cassettes. The neisserial MMEs were used to interrogate all of the other available bacterial genome sequences, revealing frequent conservation of the flanking genes combined with the presence of different gene cassettes between them. In some cases, these sites can definitively be classified as MMEs in these other genera. These findings provide additional evidence for the MME model, indicate that MME-directed investigations are a good basis for the identification of novel strain-specific genes and differences within bacterial populations and demonstrate that these elements are probably ubiquitously involved in genetic exchange, particularly in naturally competent bacteria. PMID- 17921486 TI - Spiralian phylogenomics supports the resurrection of Bryozoa comprising Ectoprocta and Entoprocta. AB - Phylogenetic analyses based on 79 ribosomal proteins of 38 metazoans, partly derived from 6 new expressed sequence tag projects for Ectoprocta, Entoprocta, Sipuncula, Annelida, and Acanthocephala, indicate the monophyly of Bryozoa comprising Ectoprocta and Entoprocta, 2 taxa that have been separated for more than a century based on seemingly profound morphological differences. Our results also show that bryozoans are more closely related to Neotrochozoa, including molluscs and annelids, than to Syndermata, the latter comprising Rotifera and Acanthocephala. Furthermore, we find evidence for the position of Sipuncula within Annelida. These findings suggest that classical developmental and morphological key characters such as cleavage pattern, coelomic cavities, gut architecture, and body segmentation are subject to greater evolutionary plasticity than traditionally assumed. PMID- 17921487 TI - A study of the coevolutionary patterns operating within the env gene of the HIV-1 group M subtypes. AB - The env gene of human immunodeficiency virus (HIV) is a functionally important gene responsible for the production of protein products (gp120 and gp41) involved in host cell recognition, binding, and entry. This occurs through a complex and, as yet, not fully understood process of protein-protein interaction and within and between protein functional communication. Exposure on the surface of active HIV virions means the gp120-gp41 complexes are subjected to intense immune system pressure and have, therefore, evolved mechanisms to avoid neutralization. Using protein-coding sequences representing all the HIV type-1 (HIV-1) group M subtypes, we have identified amino acids within the env gene whose evolution is inextricably linked over the entire HIV-1 group M epidemic. We identified 848 pairs of coevolving residues (involving 263 out of 764 amino acid sites), which represent 0.29% of all possible pairs. Of the coevolving pairs, 68% were significantly correlated by hydrophobicity, molecular weight, or both hydrophobicity and molecular weight. Subsequent grouping of coevolving pairs resulted in the identification of 290 groups of amino acid residues, with the size of these groups ranging from 2 to 10 amino acid residues. Many of these dependencies are correlated by function including CD4 binding, coreceptor binding, glycosylation, and protein-protein interaction. This analysis provides important information regarding the functional dependencies observed within all the HIV-1 group M subtypes and may assist in the identification of functional protein domains and therapeutic targets within the HIV-1 env gene. PMID- 17921488 TI - Multiple origins and rapid evolution of duplicated mitochondrial genes in parthenogenetic geckos (Heteronotia binoei; Squamata, Gekkonidae). AB - Accumulating evidence for alternative gene orders demonstrates that vertebrate mitochondrial genomes are more evolutionarily dynamic than previously thought. Several lineages of parthenogenetic lizards contain large, tandem duplications that include rRNA, tRNA, and protein-coding genes, as well as the control region. Such duplications are hypothesized as intermediate stages in gene rearrangement, but the early stages of their evolution have not been previously studied. To better understand the evolutionary dynamics of duplicated segments of mitochondrial DNA, we sequenced 10 mitochondrial genomes from recently formed ( approximately 300,000 years ago) hybrid parthenogenetic geckos of the Heteronotia binoei complex and 1 from a sexual form. These genomes included some with an arrangement typical of vertebrates and others with tandem duplications varying in size from 5.7 to 9.4 kb, each with different gene contents and duplication endpoints. These results, together with phylogenetic analyses, indicate independent and frequent origins of the duplications. Small, direct repeats at the duplication endpoints imply slipped-strand error as a mechanism generating the duplications as opposed to a false initiation/termination of DNA replication mechanism that has been invoked to explain duplications in other lizard mitochondrial systems. Despite their recent origin, there is evidence for nonfunctionalization of genes due primarily to deletions, and the observed pattern of gene disruption supports the duplication-deletion model for rearrangement of mtDNA gene order. Conversely, the accumulation of mutations between these recent duplicates provides no evidence for gene conversion, as has been reported in some other systems. These results demonstrate that, despite their long-term stasis in gene content and arrangement in some lineages, vertebrate mitochondrial genomes can be evolutionary dynamic even at short timescales. PMID- 17921489 TI - Silica in plants: biological, biochemical and chemical studies. AB - BACKGROUND: The incorporation of silica within the plant cell wall has been well documented by botanists and materials scientists; however, the means by which plants are able to transport silicon and control its polymerization, together with the roles of silica in situ, are not fully understood. RECENT PROGRESS: Recent studies into the mechanisms by which silicification proceeds have identified the following: an energy-dependent Si transporter; Si as a biologically active element triggering natural defence mechanisms; and the means by which abiotic toxicities are alleviated by silica. A full understanding of silica formation in vivo still requires an elucidation of the role played by the environment in which silica formation occurs. Results from in-vitro studies of the effects of cell-wall components associated with polymerized silica on mineral formation illustrate the interactions occurring between the biomolecules and silica, and the effects their presence has on the mineralized structures so formed. SCOPE: This Botanical Briefing describes the uptake, storage and function of Si, and discusses the role biomolecules play when incorporated into model systems of silica polymerization as well as future directions for research in this field. PMID- 17921490 TI - Using an ecophysiological analysis to dissect genetic variability and to propose an ideotype for nitrogen nutrition in pea. AB - BACKGROUNDS AND AIMS: Nitrogen nutrition of legumes, which relies both on atmospheric N2 and soil mineral N, remains a major limiting factor of growth. A decade ago, breeders tried to increase N uptake through hypernodulation. Despite their high nodule biomass, hypernodulating mutants were never shown to accumulate more nitrogen than wild types; they even generally displayed depressed shoot growth. The aim of this study was to dissect genetic variability associated with N nutrition in relation to C nutrition, using an ecophysiological framework and to propose an ideotype for N nutrition in pea. METHODS: Five pea genotypes (Pisum sativum) characterized by contrasting root and nodule biomasses were grown in the field. Variability among genotypes in dry matter and N accumulation was analysed, considering both the structures involved in N acquisition in terms of root and nodule biomass and their efficiency, in terms of N accumulated through mineral N absorption or symbiotic N2 fixation per amount of root or nodule biomass, respectively. KEY RESULTS: Nodule efficiency of hypernodulating mutants was negatively correlated to nodule biomass, presumably due to the high carbon costs induced by their excessive nodule formation. Root efficiency was only negatively correlated to root biomass before the beginning of the seed-filling stage, suggesting competition for carbon between root formation and functioning during the early stages of growth. This was no longer the case after the beginning of the seed-filling stage and nitrate absorption was then positively correlated to root biomass. CONCLUSIONS: Due to the high C costs induced by nodule formation and its detrimental effect on shoot and root growth, selecting traits for the improvement of N acquisition by legumes must be engineered (a) considering inter relationships between C and N metabolisms and (b) in terms of temporal complementarities between N2 fixation and nitrate absorption rather than through direct increase of nodule and/or root biomass. PMID- 17921491 TI - Improved BLAST searches using longer words for protein seeding. AB - MOTIVATION: The blastp and tblastn modules of BLAST are widely used methods for searching protein queries against protein and nucleotide databases, respectively. One heuristic used in BLAST is to consider only database sequences that contain a high-scoring match of length at most 5 to the query. We implemented the capability to use words of length 6 or 7. We demonstrate an improved trade-off between running time and retrieval accuracy, controlled by the score threshold used for short word matches. For example, the running time can be reduced by 20 30% while achieving ROC (receiver operator characteristic) scores similar to those obtained with current default parameters. AVAILABILITY: The option to use long words is in the NCBI C and C++ toolkit code for BLAST, starting with version 2.2.16 of blastall. A Linux executable used to produce the results herein is available at: ftp://ftp.ncbi.nlm.nih.gov/pub/agarwala/protein_longwords PMID- 17921492 TI - HMM-Kalign: a tool for generating sub-optimal HMM alignments. AB - Recent development of strategies using multiple sequence alignments (MSA) or profiles to detect remote homologies between proteins has led to a significant increase in the number of proteins whose structures can be generated by comparative modeling methods. However, prediction of the optimal alignment between these highly divergent homologous proteins remains a difficult issue. We present a tool based on a generalized Viterbi algorithm that generates optimal and sub-optimal alignments between a sequence and a Hidden Markov Model. The tool is implemented as a new function within the HMMER package called hmmkalign. PMID- 17921493 TI - BiasViz: visualization of amino acid biased regions in protein alignments. AB - About a third of all protein sequences have at least one composition biased region (CBR). Such regions might act as linkers between protein domains but often confer specific binding to various molecules; therefore, their characterization in terms of their boundaries and over-represented residues is important. Analysis of CBRs in a particular sequence can be time consuming if several types of biases have to be explored and their position visualized. Assessment of the significance of the detected CBRs can be approached by comparison to homologous protein sequences. To assist this procedure, we have developed BiasViz, a tool that allows to graphically studying local amino acid composition in protein sequences of a multiple sequence alignment. PMID- 17921494 TI - Biclustering as a method for RNA local multiple sequence alignment. AB - MOTIVATIONS: Biclustering is a clustering method that simultaneously clusters both the domain and range of a relation. A challenge in multiple sequence alignment (MSA) is that the alignment of sequences is often intended to reveal groups of conserved functional subsequences. Simultaneously, the grouping of the sequences can impact the alignment; precisely the kind of dual situation biclustering is intended to address. RESULTS: We define a representation of the MSA problem enabling the application of biclustering algorithms. We develop a computer program for local MSA, BlockMSA, that combines biclustering with divide and-conquer. BlockMSA simultaneously finds groups of similar sequences and locally aligns subsequences within them. Further alignment is accomplished by dividing both the set of sequences and their contents. The net result is both a multiple sequence alignment and a hierarchical clustering of the sequences. BlockMSA was tested on the subsets of the BRAliBase 2.1 benchmark suite that display high variability and on an extension to that suite to larger problem sizes. Also, alignments were evaluated of two large datasets of current biological interest, T box sequences and Group IC1 Introns. The results were compared with alignments computed by ClustalW, MAFFT, MUCLE and PROBCONS alignment programs using Sum of Pairs (SPS) and Consensus Count. Results for the benchmark suite are sensitive to problem size. On problems of 15 or greater sequences, BlockMSA is consistently the best. On none of the problems in the test suite are there appreciable differences in scores among BlockMSA, MAFFT and PROBCONS. On the T box sequences, BlockMSA does the most faithful job of reproducing known annotations. MAFFT and PROBCONS do not. On the Intron sequences, BlockMSA, MAFFT and MUSCLE are comparable at identifying conserved regions. AVAILABILITY: BlockMSA is implemented in Java. Source code and supplementary datasets are available at http://aug.csres.utexas.edu/msa/ PMID- 17921496 TI - Extracting three-way gene interactions from microarray data. AB - MOTIVATION: It is an important and difficult task to extract gene network information from high-throughput genomic data. A common approach is to cluster genes using pairwise correlation as a distance metric. However, pairwise correlation is clearly too simplistic to describe the complex relationships among real genes since co-expression relationships are often restricted to a specific set of biological conditions/processes. In this study, we described a three-way gene interaction model that captures the dynamic nature of co-expression relationship between a gene pair through the introduction of a controller gene. RESULTS: We surveyed 0.4 billion possible three-way interactions among 1000 genes in a microarray dataset containing 678 human cancer samples. To test the reproducibility and statistical significance of our results, we randomly split the samples into a training set and a testing set. We found that the gene triplets with the strongest interactions (i.e. with the smallest P-values from appropriate statistical tests) in the training set also had the strongest interactions in the testing set. A distinctive pattern of three-way interaction emerged from these gene triplets: depending on the third gene being expressed or not, the remaining two genes can be either co-expressed or mutually exclusive (i.e. expression of either one of them would repress the other). Such three-way interactions can exist without apparent pairwise correlations. The identified three-way interactions may constitute candidates for further experimentation using techniques such as RNA interference, so that novel gene network or pathways could be identified. PMID- 17921495 TI - Evaluation and integration of 49 genome-wide experiments and the prediction of previously unknown obesity-related genes. AB - MOTIVATION: Genome-wide experiments only rarely show resounding success in yielding genes associated with complex polygenic disorders. We evaluate 49 obesity-related genome-wide experiments with publicly available findings including microarray, genetics, proteomics and gene knock-down from human, mouse, rat and worm, in terms of their ability to rediscover a comprehensive set of genes previously found to be causally associated or having variants associated with obesity. RESULTS: Individual experiments show poor predictive ability for rediscovering known obesity-associated genes. We show that intersecting the results of experiments significantly improves the sensitivity, specificity and precision of the prediction of obesity-associated genes. We create an integrative model that statistically significantly outperforms all 49 individual genome-wide experiments. We find that genes known to be associated with obesity are significantly implicated in more obesity-related experiments and use this to provide a list of genes that we predict to have the highest likelihood of association for obesity. The approach described here can include any number and type of genome-wide experiments and might be useful for other complex polygenic disorders as well. PMID- 17921497 TI - Four-body scoring function for mutagenesis. AB - MOTIVATION: There is a need for an efficient and accurate computational method to identify the effects of single- and multiple-residue mutations on the stability and reactivity of proteins. Such a method should ideally be consistent and yet applicable in a widespread manner, i.e. it should be applied to various proteins under the same parameter settings, and have good predictive power for all of them. RESULTS: We develop a Delaunay tessellation-based four-body scoring function to predict the effects of single- and multiple-residue mutations on the stability and reactivity of proteins. We test our scoring function on sets of single-point mutations used by several previous studies. We also assemble a new, diverse set of 237 single- and multiple-residue mutations, from over 24 different publications. The four-body scoring function correctly predicted the changes to the stability of 169 out of 210 mutants (80.5%), and the changes to the reactivity of 17 out of 27 mutants (63%). For the mutants that had the changes in stability/reactivity quantified (using reaction rates, temperatures, etc.), an average Spearman rank correlation coefficient of 0.67 was achieved with the four body scores. We also develop an efficient method for screening huge numbers of mutants of a protein, called combinatorial mutagenesis. In one study, 64 million mutants of a cold-shock nucleus binding domain protein 1CSQ, with six of its residues being changed to all possible (20) amino acids, were screened within a few hours on a PC, and all five stabilizing mutants reported were correctly identified as stabilizing by combinatorial mutagenesis. PMID- 17921498 TI - coliSNP database server mapping nsSNPs on protein structures. AB - We have developed coliSNP, a database server (http://yayoi.kansai.jaea.go.jp/colisnp) that maps non-synonymous single nucleotide polymorphisms (nsSNPs) on the three-dimensional (3D) structure of proteins. Once a week, the SNP data from the dbSNP database and the protein structure data from the Protein Data Bank (PDB) are downloaded, and the correspondence of the two data sets is automatically tabulated in the coliSNP database. Given an amino acid sequence, protein name or PDB ID, the server will immediately provide known nsSNP information, including the amino acid mutation caused by the nsSNP, the solvent accessibility, the secondary structure and the flanking residues of the mutated residue in a single page. The position of the nsSNP within the amino acid sequence and on the 3D structure of the protein can also be observed. The database provides key information with which to judge whether an observed nsSNP critically affects protein function and/or stability. As far as we know, this is the only web-based nsSNP database that automatically compiles SNP and protein information in a concise manner. PMID- 17921499 TI - RNA FRABASE version 1.0: an engine with a database to search for the three dimensional fragments within RNA structures. AB - The RNA FRABASE is a web-accessible engine with a relational database, which allows for the automatic search of user-defined, 3D RNA fragments within a set of RNA structures. This is a new tool to search and analyse RNA structures, directed at the 3D structure modelling. The user needs to input either RNA sequence(s) and/or secondary structure(s) given in a 'dot-bracket' notation. The algorithm searching for the requested 3D RNA fragments is very efficient. As of August 2007, the database contains: (i) RNA sequences and secondary structures, in the 'dot-bracket' notation, derived from 1065 protein data bank (PDB)-deposited RNA structures and their complexes, (ii) a collection of atom coordinates of unmodified and modified nucleotide residues occurring in RNA structures, (iii) calculated RNA torsion angles and sugar pucker parameters and (iv) information about base pairs. Advanced query involves filters sensitive to: modified residue contents, experimental method used and limits of conformational parameters. The output list of query-matching RNA fragments gives access to their coordinates in the PDB-format files, ready for direct download and visualization, conformational parameters and information about base pairs. The RNA FRABASE is automatically, monthly updated and is freely accessible at http://rnafrabase.ibch.poznan.pl (mirror at http://cerber.cs.put.poznan.pl/rnadb). PMID- 17921500 TI - The Sulfolobus solfataricus radA paralogue sso0777 is DNA damage inducible and positively regulated by the Sta1 protein. AB - Little is known about the regulation of the DNA damage-mediated gene expression in archaea. Here we report that the addition of actinomycin D to Sulfolobus solfataricus cultures triggers the expression of the radA paralogue sso0777. Furthermore, a specific retarded band is observed when electrophoretic mobility shift assays (EMSAs) with crude S. solfataricus cell extracts and the sso0777 promoter were carried out. The protein that binds to this promoter was isolated and identified as Sta1. Footprinting experiments have shown that the Sta1 DNA binding site is included in the ATTTTTTATTTTCACATGTAAGATGTTTATT sequence, which is located upstream the putative TTG translation starting codon of the sso0777 gene. Additionally, gel electrophoretic mobility retardation experiments using mutant sso0777 promoter derivatives show the presence of three essential motifs (TTATT, CANGNA and TTATT) that are absolutely required for Sta1 DNA binding. Finally, in vitro transcription experiments confirm that Sta1 functions as an activator for sso0777 gene expression being the first identified archaeal regulatory protein associated with the DNA damage-mediated induction of gene expression. PMID- 17921501 TI - Targeted inhibition of the hepatitis C internal ribosomal entry site genomic RNA with oligonucleotide conjugates. AB - Hepatitis C is a major public health concern, with an estimated 170 million people infected worldwide and an urgent need for new drug development. An attractive therapeutic approach is to prevent the 'cap-independent' translation initiation of the viral proteins by interfering with both the structure and function of the hepatitis C viral internal ribosomal entry site (HCV IRES). Towards this goal, we report the design, synthesis and purification of novel bi functional molecules containing DNA or RNA antisenses attached to functional groups performing RNA hydrolysis. These 5' or 3'-coupled conjugates bind the HCV IRES with affinity and specificity and elicit targeted hydrolysis of the viral genomic RNA after short (1 h) incubation at low (500 nM) concentration at 37 degrees C in vitro. Additional secondary cleavage sites are induced and their mapping within the RNA structure indicates that functional domains IIIb-e are excised from the IRES that, based on cryo-EM studies, becomes incapable of binding the small ribosomal subunit and initiation factor 3 (eIF3). All these molecules inhibit, in a dose-dependent manner, the 'IRES-dependent' translation in vitro. The 5'-coupled imidazole conjugate reduces viral protein synthesis by half at a 300 nM concentration (IC50), corresponding to a 4-fold increase of activity when compared to the naked oligonucleotide. These new conjugates are now being tested for activity on infected hepatic cell lines. PMID- 17921502 TI - TOPDB: topology data bank of transmembrane proteins. AB - The Topology Data Bank of Transmembrane Proteins (TOPDB) is the most complete and comprehensive collection of transmembrane protein datasets containing experimentally derived topology information currently available. It contains information gathered from the literature and from public databases available on the internet for more than a thousand transmembrane proteins. TOPDB collects details of various experiments that were carried out to learn about the topology of particular transmembrane proteins. In addition to experimental data from the literature, an extensive collection of structural data was also compiled from PDB and from PDBTM. Because topology information is often incomplete, for each protein in the database the most probable topology that is consistent with the collected experimental constraints was also calculated using the HMMTOP transmembrane topology prediction algorithm. Each record in TOPDB also contains information on the given protein sequence, name, organism and cross references to various other databases. The web interface of TOPDB includes tools for searching, relational querying and data browsing as well as for visualization. TOPDB is designed to bridge the gap between the number of transmembrane proteins available in sequence databases and the publicly accessible topology information of experimentally or computationally studied transmembrane proteins. TOPDB is available at http://topdb.enzim.hu. PMID- 17921504 TI - Complications of open abdominal aortic surgery: the endovascular solution. AB - OBJECTIVES: Aorto-enteric fistulas (AEFs) and para-anastomotic aneurysms (PAAs) are uncommon complications of open aortic surgery (0.5-2.5%) and (0.2-15%), respectively. AEF if untreated is often fatal and surgical management is associated with mortality up to 90%. The risk of PAA rupture carries a mortality of 58%. We present our experience in ten patients with the endovascular treatment of these complications to define the role of endovascular repair in this high risk group. METHODS: This is a retrospective review of patients presenting acutely with complications of open aortic surgery. From January 2003 to March 2006, ten patients, all males with a mean age of 73 years presented through the Accident and Emergency department and were assessed with contrast enhanced CT. Five patients presented with secondary AEFs. Three patients with PAAs. Another patient presented with a secondary mycotic aneurysm of the thoracic aorta following open repair of abdominal aortic aneurysm and finally a patient with a femoral pseudoaneurysm. The mean time from the original procedure to presentation was 50 months. All patients were offered endovascular management after stabilisation as they were deemed as high-risk surgical patients. RESULTS: No intra-procedural complications were recorded. The in-hospital 30-day mortality was 1 (10%) patient due to multiple organ failure. One patient died six months later due to an unrelated event. The average in-hospital stay was 5.4 days; median follow-up period was 28 months. All patients were repeatedly admitted after discharge due to septic episodes for which they received IV antibiotics. Repeated cultures for all patients were only positive on four occasions. CONCLUSION: Endovascular stent-graft repair of AEF and PAAs is a viable alternative to open surgery. It is likely to be associated with less mortality and morbidity and in-hospital stay. It should be considered as an alternative in high-risk patients. Persistence of the infection remains a problem, however, in our experience; it can be well controlled through long-term antibiotics. PMID- 17921503 TI - Discovery of Fur binding site clusters in Escherichia coli by information theory models. AB - Fur is a DNA binding protein that represses bacterial iron uptake systems. Eleven footprinted Escherichia coli Fur binding sites were used to create an initial information theory model of Fur binding, which was then refined by adding 13 experimentally confirmed sites. When the refined model was scanned across all available footprinted sequences, sequence walkers, which are visual depictions of predicted binding sites, frequently appeared in clusters that fit the footprints ( approximately 83% coverage). This indicated that the model can accurately predict Fur binding. Within the clusters, individual walkers were separated from their neighbors by exactly 3 or 6 bases, consistent with models in which Fur dimers bind on different faces of the DNA helix. When the E. coli genome was scanned, we found 363 unique clusters, which includes all known Fur-repressed genes that are involved in iron metabolism. In contrast, only a few of the known Fur-activated genes have predicted Fur binding sites at their promoters. These observations suggest that Fur is either a direct repressor or an indirect activator. The Pseudomonas aeruginosa and Bacillus subtilis Fur models are highly similar to the E. coli Fur model, suggesting that the Fur-DNA recognition mechanism may be conserved for even distantly related bacteria. PMID- 17921505 TI - Thermal coronary angiography in pediatric coronary artery bypass grafting. AB - A 2-year-old boy was referred to our institution for treatment of coronary artery obstruction after an arterial switch operation (ASO). We performed a pediatric coronary artery bypass graft (CABG) with skeletonized left internal thoracic artery (LITA). The patient was 88 cm tall and weighed 11 kg. The left anterior descending (LAD) coronary artery was also identified about 1 mm in diameter. Anastomotic failure of imposed stricture was detected with an infrared camera (IRIS-V, VERITAS, Tokyo, Japan). In the situation of CABG, thermal coronary angiography (TCA) is very useful to reveal satisfactory blood flow into the left coronary system during this procedure. PMID- 17921506 TI - An antisense transcript spanning the CGG repeat region of FMR1 is upregulated in premutation carriers but silenced in full mutation individuals. AB - Expansion of the polymorphic CGG repeats within the 5'-UTR of the FMR1 gene is associated with variable transcriptional regulation of FMR1. Here we report a novel gene, ASFMR1, overlapping the CGG repeat region of FMR1 and transcribed in the antisense orientation. The ASFMR1 transcript is spliced, polyadenylated and exported to the cytoplasm. Similar to FMR1, ASFMR1 is upregulated in individuals with premutation alleles and is not expressed from full mutation alleles. Moreover, it exhibits premutation-specific alternative splicing. Taken together, these observations suggest that in addition to FMR1, ASFMR1 may contribute to the variable phenotypes associated with the CGG repeat expansion. PMID- 17921507 TI - The grainyhead like 2 gene (GRHL2), alias TFCP2L3, is associated with age-related hearing impairment. AB - Age-related hearing impairment (ARHI) is the most prevalent sensory impairment in the elderly. ARHI is a complex disease caused by an interaction between environmental and genetic factors. The contribution of various environmental factors has been relatively extensively studied. In contrast, investigations to identify the genetic risk factors have only recently been initiated. In this paper we describe the results of an association study performed on 2418 ARHI samples derived from nine centers from seven European countries. In 70 candidate genes, a total of 768 tag single nucleotide polymorphisms (SNPs) were selected based on HAPMAP data. These genes were chosen among the monogenic hearing loss genes identified in mice and men in addition to several strong functional candidates. After genotyping and data polishing, statistical analysis of all samples combined resulted in a P-value that survived correction for multiple testing for one SNP in the GRHL2 gene. Other SNPs in this gene were also associated, albeit to a lesser degree. Subsequently, an analysis of the most significant GRHL2 SNP was performed separately for each center. The direction of the association was identical in all nine centers. Two centers showed significant associations and a third center showed a trend towards significance. Subsequent fine mapping of this locus demonstrated that the majority of the associated SNPs reside in intron 1. We hypothesize that the causative variant may change the expression levels of a GRHL2 isoform. PMID- 17921508 TI - The computation of ICRP dose coefficients for intakes of radionuclides with PLEIADES: biokinetic aspects. AB - The ICRP has published dose coefficients for the ingestion or inhalation of radionuclides in a series of reports covering intakes by workers and members of the public including children and pregnant or lactating women. The calculation of these coefficients conveniently divides into two distinct parts--the biokinetic and dosimetric. This paper gives a brief summary of the methods used to solve the biokinetic problem in the generation of dose coefficients on behalf of the ICRP, as implemented in the Health Protection Agency's internal dosimetry code PLEIADES. PMID- 17921509 TI - Automated detection of irradiated food with the comet assay. AB - Food irradiation is the process of exposing food to ionising radiation in order to disinfect, sanitise, sterilise and preserve food or to provide insect disinfestation. Irradiated food should be adequately labelled according to international and national guidelines. In many countries, there are furthermore restrictions to the product-specific maximal dose that can be administered. Therefore, there is a need for methods that allow detection of irradiated food, as well as for methods that provide a reliable dose estimate. In recent years, the comet assay was proposed as a simple, rapid and inexpensive method to fulfil these goals, but further research is required to explore the full potential of this method. In this paper we describe the use of an automated image analysing system to measure DNA comets which allow the discrimination between irradiated and non-irradiated food as well as the set-up of standard dose-response curves, and hence a sufficiently accurate dose estimation. PMID- 17921510 TI - Determination of natural radioactivity in drinking water in private dug wells in Akure, Southwestern Nigeria. AB - Gamma spectrometry of well water was done to measure the activity concentrations of gamma-emitters 226Ra and 228Ra from 238U and 232Th series, respectively, and 40K in their waters. Measurements were done using high-purity (HPGe) co-axial detectors (Canberra, GC 2018-7500 ) coupled to Canberra Multichannel Analyser system. Activity concentrations ranged from 0.57 to 26.86, 0.20 to 60.06 and 0.35 to 29.01 Bq l(-1) for 226Ra, 228Ra and 40K, respectively. Activity concentrations were compared with data from other parts of the world and used to estimate annual effective dose for three age groups. Total annual effective doses from the intake of these radionuclides in the waters ranged from 0.02 to 76.84, 0.02 to 38.80 and 0.05 to 481.60 mSv y(-1) for age group <1, 2-7 and > or =17 y, respectively. These values are considerably higher than both the World Health Organisation and the International Commission on Radiological Protection limits. PMID- 17921511 TI - Radionuclide content and associated radiation hazards of building materials and by-products in Baoji, West China. AB - Seven types of common building materials and by-products of coal-fired power plants collected from Baoji, West China, were analysed for the natural radioactivity of 226Ra, 232Th and 40K using gamma ray spectrometry with an NaI(Tl) detector. The average activity concentrations vary from 23.0 to 112.2, 20.2 to 147.5 and 113.2 to 890.8 Bq kg(-1) for 226Ra, 232Th and 40K, respectively. The measured activity concentrations for these radionuclides were compared with the data reported from other countries and with the worldwide average activity of soil. As a measure of radiation hazard to the people, the radium equivalent activities, total annual effective dose and activity concentration index were estimated. The radium equivalent activities of the studied samples are below the internationally accepted values. The calculated total annual effective dose and the activity concentration index of seven types of common building materials are <1 mSv y(-1) and 1, respectively. But fly ash and bottom ash exhibit the higher values that exceed and be close to the acceptable values, respectively. This study shows that the measured building materials do not pose any significant source of radiation hazard and are safe for use in the construction of dwellings. Nevertheless, when fly ash and bottom ash are used in dwelling construction, it is important to assess their radiation potential. PMID- 17921512 TI - Patient dosimetry in interventional cardiology at the University Hospital of Osijek. AB - The interventional cardiology was recently implemented at the University Hospital of Osijek. Patients' absorbed doses during coronary angiography (CA) and the percutaneous transluminal coronary angioplasty (PTCA) procedures were measured and compared with published data and international standards. All patients undergoing CA or PTCA procedures during a 1-month period were included in the study. Patients' doses are expressed in terms of dose area product (DAP) per procedure. The patients' DAPs ranged from 2.6 to 210 Gy cm2 (average of 59 Gy cm2) during CAs, and from 61 to 220 Gy cm2 (average of 120 Gy cm2) during PTCAs. Patients' doses during CAs and PTCAs at the University Hospital of Osijek are in good agreement with the published ones. In complex cases, the radiochromic dosimetry films were used to show possible dose distributions across patient's skin. The film dosimetry showed a limitation of using only DAP values for the estimation of skin injuries risk. PMID- 17921513 TI - Can calcimimetics inhibit nodular hyperplasia of parathyroid glands? PMID- 17921514 TI - Comment on EBPG guideline on dialysis strategies. PMID- 17921515 TI - A memorable experience. AB - The electrocardiographic phenomenon of cardiac memory results from aberrant ventricular activation and causes transient ECG features that can mimic ischaemia. We present the case of a patient with ischaemic cardiomyopathy who exhibited evidence of T-wave memory in the setting of multiple episodes of non sustained ventricular tachycardia that were triggered by an insect bite. PMID- 17921516 TI - Giant left ventricular pseudoaneurysm: a rare complication following left ventricular rupture caused by myocardial infarction. PMID- 17921518 TI - Cortical Aerenchyma formation in hypocotyl and adventitious roots of Luffa cylindrica subjected to soil flooding. AB - BACKGROUND AND AIMS: Aerenchyma formation is thought to be one of the important morphological adaptations to hypoxic stress. Although sponge gourd is an annual vegetable upland crop, in response to flooding the hypocotyl and newly formed adventitious roots create aerenchyma that is neither schizogenous nor lysigenous, but is produced by radial elongation of cortical cells. The aim of this study is to characterize the morphological changes in flooded tissues and the pattern of cortical aerenchyma formation, and to analyse the relative amount of aerenchyma formed. METHODS: Plants were harvested at 16 d after the flooding treatment was initiated. The root system was observed, and sections of fresh materials (hypocotyl, tap root and adventitious root) were viewed with a light or fluorescence microscope. Distributions of porosity along adventitious roots were estimated by a pycnometer method. KEY RESULTS: Under flooded conditions, a considerable part of the root system consisted of new adventitious roots which soon emerged and grew quickly over the soil surface. The outer cortical cells of these roots and those of the hypocotyl elongated radially and contributed to the development of large intercellular spaces. The elongated cortical cells of adventitious roots were clearly T-shaped, and occurred regularly in mesh-like lacunate structures. In these positions, slits were formed in the epidermis. In the roots, the enlargement of the gas space system began close to the apex in the cortical cell layers immediately beneath the epidermis. The porosity along these roots was 11-45 %. In non-flooded plants, adventitious roots were not formed and no aerenchyma developed in the hypocotyl or tap root. CONCLUSIONS: Sponge gourd aerenchyma is produced by the unique radial elongation of cells that make the expansigeny. These morphological changes seem to enhance flooding tolerance by promoting tissue gas exchange, and sponge gourd might thereby adapt to flooding stress. PMID- 17921517 TI - Proteomics approach to identify dehydration responsive nuclear proteins from chickpea (Cicer arietinum L.). AB - Dehydration or water-deficit is one of the most important environmental stress factors that greatly influences plant growth and development and limits crop productivity. Plants respond and adapt to such stress by altering their cellular metabolism and activating various defense machineries. Mechanisms that operate signal perception, transduction, and downstream regulatory events provide valuable information about the underlying pathways involved in environmental stress responses. The nuclear proteins constitute a highly organized, complex network that plays diverse roles during cellular development and other physiological processes. To gain a better understanding of dehydration response in plants, we have developed a comparative nuclear proteome in a food legume, chickpea (Cicer arietinum L.). Three-week-old chickpea seedlings were subjected to progressive dehydration by withdrawing water and the changes in the nuclear proteome were examined using two-dimensional gel electrophoresis. Approximately 205 protein spots were found to be differentially regulated under dehydration. Mass spectrometry analysis allowed the identification of 147 differentially expressed proteins, presumably involved in a variety of functions including gene transcription and replication, molecular chaperones, cell signaling, and chromatin remodeling. The dehydration responsive nuclear proteome of chickpea revealed a coordinated response, which involves both the regulatory as well as the functional proteins. This study, for the first time, provides an insight into the complex metabolic network operating in the nucleus during dehydration. PMID- 17921519 TI - Association of the gastric alcohol dehydrogenase gene ADH7 with variation in alcohol metabolism. AB - Seven alcohol-metabolizing enzymes are encoded by the human alcohol dehydrogenase (ADH) gene cluster on chromosome 4q22-23. One of these genes, ADH7, is uniquely expressed in the stomach mucosa and can influence metabolism of alcohol before its absorption into the blood. However, the contribution of ADH7 to the overall genetic variation in alcohol oxidation in vivo is unknown. Data on in vivo alcohol metabolism were obtained for 206 Australian twin pairs of Caucasian ancestry, following ingestion of a standard dose (0.75 g kg(-1) body weight) of alcohol. Twenty-five single nucleotide polymorphisms that cover the ADH7 encoding region were genotyped. The patterns of linkage disequilibrium among these SNPs identified a recombinational hotspot within intron 7 of the ADH7 gene. A model for the absorption and elimination of alcohol from the body led to the identification of haplotypes associated with inter-individual variation in the early stages of alcohol metabolism. These are within a 35 kb DNA tract contained in the region 5' of intron 7 in the ADH7 gene. The region accounts for 18% of the linkage for alcohol concentration associated with the ADH region, or approximately 11% of the genetic variance. PMID- 17921520 TI - A rational mechanism for combination treatment of Huntington's disease using lithium and rapamycin. AB - Huntington's disease (HD) is caused by a polyglutamine expansion mutation in the huntingtin protein that confers a toxic gain-of-function and causes the protein to become aggregate-prone. Aggregate-prone proteins are cleared by macroautophagy, and upregulating this process by rapamycin, which inhibits the mammalian target of rapamycin (mTOR), attenuates their toxicity in various HD models. Recently, we demonstrated that lithium induces mTOR-independent autophagy by inhibiting inositol monophosphatase (IMPase) and reducing inositol and IP3 levels. Here we show that glycogen synthase kinase-3beta (GSK-3beta), another enzyme inhibited by lithium, has opposite effects. In contrast to IMPase inhibition that enhances autophagy, GSK3beta inhibition attenuates autophagy and mutant huntingtin clearance by activating mTOR. In order to counteract the autophagy inhibitory effects of mTOR activation resulting from lithium treatment, we have used the mTOR inhibitor rapamycin in combination with lithium. This combination enhances macroautophagy by mTOR-independent (IMPase inhibition by lithium) and mTOR-dependent (mTOR inhibition by rapamycin) pathways. We provide proof-of-principle for this rational combination treatment approach in vivo by showing greater protection against neurodegeneration in an HD fly model with TOR inhibition and lithium, or in HD flies treated with rapamycin and lithium, compared with either pathway alone. PMID- 17921521 TI - Neurofurans, novel indices of oxidant stress derived from docosahexaenoic acid. AB - Isoeicosanoids are free radical-catalyzed isomers of the enzymatic products of arachidonic acid. They are formed in situ in cell membranes, are cleaved, circulate, and are excreted in urine. Isomers of prostaglandin F(2alpha), the F(2)-isoprostanes, have emerged as sensitive indices of lipid peroxidation in vivo. Analogous compounds formed from docosahexaenoic acid (DHA) are termed neuroprostanes and are more abundant than isoprostanes (iPs) in brain. Isofurans are another class of isoeicosanoids characterized by a substituted tetrahydrofuran ring. They are preferentially formed, relative to iPs, under conditions of elevated oxygen tension. Here, we report the discovery of neurofurans (nFs), the analogous family of compounds formed from DHA. Formation of nFs is characterized by mass spectrometry and confirmed by oxidation of DHA in vitro and following CCl(4) administration in liver in vivo. It is demonstrated that the levels of nFs are elevated in the brain cortex of a mouse model of Alzheimer disease and are depressed in mouse brain cortex by deletion of p47(phox), an essential component of the phagocyte NADPH oxidase. Measurement of the nFs may ultimately prove useful in diagnosis, timing, and selection of dose in the treatment and chemoprevention of neurodegenerative disease. PMID- 17921523 TI - Isolation and characterization of arbuscules from roots of an increased-arbuscule forming mutant of Lotus japonicus. AB - BACKGROUND AND AIMS: Previous methods for isolation of arbuscules from mycorrhizal roots are time-consuming, complex and expensive. Therefore, a simple, rapid and inexpensive method for the isolation of metabolically active arbuscules from plant root of an increased-arbuscule-forming mutant of Lotus japonicus (Ljsym78-2) is described. METHODS: Roots of the L. japonicus mutant plants Ljsym78-2 colonized by Glomus sp. were separated from soil, washed with water, immersed in CaSO(4) before being cut into 5-mm pieces and homogenized with a Waring blender at 6000 rpm for 30 s. The arbuscules were purified by separation from plant tissues with a 50-mum nylon mesh, finally collecting on a 30-mum nylon mesh. Enzyme histochemical staining showed that the collected arbuscules had succinate dehydrogenase, alkaline phosphatase and acid phosphatase activities. KEY RESULTS AND CONCLUSIONS: The enzymic activity of the arbuscules was not affected after the isolation process. The establishment of this simple, rapid and inexpensive method for the isolation of metabolically active arbuscules will be useful to clarify the biochemical processes occurring in nutrient exchange at the arbuscular interface. PMID- 17921522 TI - A genome-wide survey and systematic RNAi-based characterization of helicase-like genes in Caenorhabditis elegans. AB - Helicase-like proteins play a crucial role in nucleic acid- and chromatin mediated reactions. In this study, we identified 134 helicase-like proteins in the nematode Caenorhabditis elegans and classified the proteins into 10 known subfamilies and a group of orphan genes on the basis of sequence similarity. We characterized loss-of-function phenotypes in RNA interference (RNAi)-treated animals for helicase family members, using the RNAi feeding method, and found several previously unreported phenotypes. Fifty-one (39.5%) of 129 genes tested showed development- or growth-defect phenotypes, and many of these genes were putative nematode homologs of essential genes in a unicellular eukaryote, budding yeast, suggesting conservation of these essential proteins in both species. Comparative analyses between these species identified evolutionarily diverged nematode proteins as well as conserved family members. Chromosome mapping of the nematode genes revealed 10 pairs of putative duplicated genes and clusters of C. elegans-specific SNF2-like genes and Helitrons. Analyses of transcriptional profile data revealed a predominantly oogenesis- and germline-enriched expression of many helicase-like genes. Finally, we identified the D2005.5(drh-3) gene in an RNAi-based screen for genes involved in resistance to X-ray irradiation. Analysis of DRH-3 will clarify the potentially novel mechanism by which it protects against X-ray-induced damage in C. elegans. PMID- 17921524 TI - A system for modelling cell-cell interactions during plant morphogenesis. AB - BACKGROUND AND AIMS: During the development of multicellular organisms, cells are capable of interacting with each other through a range of biological and physical mechanisms. A description of these networks of cell-cell interactions is essential for an understanding of how cellular activity is co-ordinated in regionalized functional entities such as tissues or organs. The difficulty of experimenting on living tissues has been a major limitation to describing such systems, and computer modelling appears particularly helpful to characterize the behaviour of multicellular systems. The experimental difficulties inherent to the multitude of parallel interactions that underlie cellular morphogenesis have led to the need for computer models. METHODS: A new generic model of plant cellular morphogenesis is described that expresses interactions amongst cellular entities explicitly: the plant is described as a multi-scale structure, and interactions between distinct entities is established through a topological neighbourhood. Tissues are represented as 2D biphasic systems where the cell wall responds to turgor pressure through a viscous yielding of the cell wall. KEY RESULTS: This principle was used in the development of the CellModeller software, a generic tool dedicated to the analysis and modelling of plant morphogenesis. The system was applied to three contrasting study cases illustrating genetic, hormonal and mechanical factors involved in plant morphogenesis. CONCLUSIONS: Plant morphogenesis is fundamentally a cellular process and the CellModeller software, through its underlying generic model, provides an advanced research tool to analyse coupled physical and biological morphogenetic mechanisms. PMID- 17921525 TI - Parameter optimization and field validation of the functional-structural model GREENLAB for maize at different population densities. AB - BACKGROUND AND AIMS: Plant population density (PPD) influences plant growth greatly. Functional-structural plant models such as GREENLAB can be used to simulate plant development and growth and PPD effects on plant functioning and architectural behaviour can be investigated. This study aims to evaluate the ability of GREENLAB to predict maize growth and development at different PPDs. METHODS: Two field experiments were conducted on irrigated fields in the North China Plain with a block design of four replications. Each experiment included three PPDs: 2.8, 5.6 and 11.1 plants m(-2). Detailed observations were made on the dimensions and fresh biomass of above-ground plant organs for each phytomer throughout the seasons. Growth stage-specific target files (a description of plant organ weight and dimension according to plant topological structure) were established from the measured data required for GREENLAB parameterization. Parameter optimization was conducted using a generalized least square method for the entire growth cycles for all PPDs and years. Data from in situ plant digitization were used to establish geometrical symbol files for organs that were then applied to translate model output directly into 3-D representation for each time step of the model execution. KEY RESULTS: The analysis indicated that the parameter values of organ sink variation function, and the values of most of the relative sink strength parameters varied little among years and PPDs, but the biomass production parameter, computed plant projection surface and internode relative sink strength varied with PPD. Simulations of maize plant growth based on the fitted parameters were reasonably good as indicated by the linearity and slopes similar to unity for the comparison of simulated and observed values. Based on the parameter values fitted from different PPDs, shoot (including vegetative and reproductive parts of the plant) and cob fresh biomass for other PPDs were simulated. Three-dimensional representation of individual plant and plant stand from the model output with two contrasting PPDs were presented with which the PPD effect on plant growth can be easily recognized. CONCLUSIONS: This study showed that GREENLAB model has the ability to capture plant plasticity induced by PPD. The relatively stable parameter values strengthened the hypothesis that one set of equations can govern dynamic organ growth. With further validation, this model can be used for agronomic applications such as yield optimization. PMID- 17921526 TI - Genome size variation and species relationships in Hieracium sub-genus Pilosella (Asteraceae) as inferred by flow cytometry. AB - BACKGROUND AND AIMS: Hieracium sub-genus Pilosella (hawkweeds) is a taxonomically complicated group of vascular plants, the structure of which is substantially influenced by frequent interspecific hybridization and polyploidization. Two kinds of species, 'basic' and 'intermediate' (i.e. hybridogenous), are usually recognized. In this study, genome size variation was investigated in a representative set of Central European hawkweeds in order to assess the value of such a data set for species delineation and inference of evolutionary relationships. METHODS: Holoploid and monoploid genome sizes (C- and Cx-values) were determined using propidium iodide flow cytometry for 376 homogeneously cultivated individuals of Hieracium sub-genus Pilosella, including 24 species (271 individuals), five recent natural hybrids (seven individuals) and experimental F(1) hybrids from four parental combinations (98 individuals). Chromosome counts were available for more than half of the plant accessions. Base composition (proportion of AT/GC bases) was cytometrically estimated in 73 individuals. KEY RESULTS: Seven different ploidy levels (2x-8x) were detected, with intraspecific ploidy polymorphism (up to four different cytotypes) occurring in 11 wild species. Mean 2C-values varied approx. 4.3-fold from 3.53 pg in diploid H. hoppeanum to 15.30 pg in octoploid H. brachiatum. 1Cx-values ranged from 1.72 pg in H. pilosella to 2.16 pg in H. echioides (1.26-fold). The DNA content of (high) polyploids was usually proportional to the DNA values of their diploid/low polyploid counterparts, indicating lack of processes altering genome size (i.e. genome down-sizing). Most species showed constant nuclear DNA amounts, exceptions being three hybridogenous taxa, in which introgressive hybridization was suggested as a presumable trigger for genome size variation. Monoploid genome sizes of hybridogenous species were always between the corresponding values of their putative parents. In addition, there was a good congruency between actual DNA estimates and theoretical values inferred from putative parental combinations and between DNA values of experimental F(1) hybrids and corresponding established hybridogenous taxa. CONCLUSIONS: Significant differences in genome size between hawkweed species from hybridogenous lineages involving the small-genome H. pilosella document the usefulness of nuclear DNA content as a supportive marker for reliable delineation of several of the most problematic taxa in Hieracium sub genus Pilosella (including classification of borderline morphotypes). In addition, genome size data were shown to have a good predictive value for inferring evolutionary relationships and genome constitution (i.e. putative parental combinations) in hybridogenous species. PMID- 17921527 TI - Distribution and Translocation of 141Ce (III) in Horseradish. AB - BACKGROUND AND AIMS: Rare earth elements (REEs) are used in agriculture and a large amount of them contaminate the environment and enter foods. The distribution and translocation of (141)Ce (III) in horseradish was investigated in order to help understand the biochemical behaviour and toxic mechanism of REEs in plants. METHODS: The distribution and translocation of (141)Ce (III) in horseradish were investigated using autoradiography, liquid scintillation counting (LSC) and electron microscopic autoradiography (EMARG) techniques. The contents of (141)Ce (III) and nutrient elements were analysed using an inductively coupled plasma-atomic emission spectrometer (ICP-AES). RESULTS: The results from autoradiography and LSC indicated that (141)Ce (III) could be absorbed by horseradish and transferred from the leaf to the leaf-stalk and then to the root. The content of (141)Ce (III) in different parts of horseradish was as follows: root > leaf-stalk > leaf. The uptake rates of (141)Ce (III) in horseradish changed with the different organs and time. The content of (141)Ce (III) in developing leaves was greater than that in mature leaves. The results from EMARG indicated that (141)Ce (III) could penetrate through the cell membrane and enter the mesophyll cells, being present in both extra- and intra-cellular deposits. The contents of macronutrients in horseradish were decreased by (141)Ce (III) treatment. CONCLUSIONS: (141)Ce (III) can be absorbed and transferred between organs of horseradish with time, and the distribution was found to be different at different growth stages. (141)Ce (III) can enter the mesophyll cells via apoplast and symplast channels or via plasmodesmata. (141)Ce (III) can disturb the metabolism of macronutrients in horseradish. PMID- 17921528 TI - Isoprene emission from plants: why and how. AB - BACKGROUND: Some, but not all, plants emit isoprene. Emission of the related monoterpenes is more universal among plants, but the amount of isoprene emitted from plants dominates the biosphere-atmosphere hydrocarbon exchange. SCOPE: The emission of isoprene from plants affects atmospheric chemistry. Isoprene reacts very rapidly with hydroxyl radicals in the atmosphere making hydroperoxides that can enhance ozone formation. Aerosol formation in the atmosphere may also be influenced by biogenic isoprene. Plants that emit isoprene are better able to tolerate sunlight-induced rapid heating of leaves (heat flecks). They also tolerate ozone and other reactive oxygen species better than non-emitting plants. Expression of the isoprene synthase gene can account for control of isoprene emission capacity as leaves expand. The emission capacity of fully expanded leaves varies through the season but the biochemical control of capacity of mature leaves appears to be at several different points in isoprene metabolism. CONCLUSIONS: The capacity for isoprene emission evolved many times in plants, probably as a mechanism for coping with heat flecks. It also confers tolerance of reactive oxygen species. It is an example of isoprenoids enhancing membrane function, although the mechanism is likely to be different from that of sterols. Understanding the regulation of isoprene emission is advancing rapidly now that the pathway that provides the substrate is known. PMID- 17921530 TI - Exercise training and baroreflex sensitivity in patients with neurally mediated syncope. PMID- 17921529 TI - A pathophysiologic study of tako-tsubo cardiomyopathy with F-18 fluorodeoxyglucose positron emission tomography. AB - AIMS; Our study aims to investigate the pathophysiologic mechanism underlying tako-tsubo cardiomyopathy using F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET). METHODS AND RESULTS: Fifteen patients with tako-tsubo cardiomyopathy were enrolled in this study. Plasma catecholamines, cardiac troponin T (cTnT), and D-dimer were serially evaluated in all patients. Thallium 201 ((201)Tl) single-photon emission computed tomography (SPECT) and F-18 FDG PET were performed in 10 and eight patients, respectively. Emotional or physical stress occurred in 12 (80.0%) patients. ST-T segment abnormalities existed in all patients. Thirteen patients exhibited mildly elevated cTnT, although coronary angiography did not reveal significant stenosis in any patient. Endomyocardial biopsy specimens (n = 9) demonstrated contraction-band necrosis (n = 4) and mononuclear cell infiltration (n = 3). The levels of norepinephrine and epinephrine peaked on admission (744 +/- 452 and 140 +/- 166 pg/mL, respectively). There was severely reduced uptake at the apex on F-18 FDG PET image, despite slightly reduced uptake of (201)Tl. Elevation of D-dimer was observed in nine patients. CONCLUSION: The extent of metabolic defect involving apical akinetic area was more severe than perfusion abnormality. Our data suggest that sudden emotional or physical stress may cause a catecholamine-induced metabolic disorder in the myocardium, which is probably central to this syndrome. PMID- 17921531 TI - Randomized trial of rapamycin- and paclitaxel-eluting stents with identical biodegradable polymeric coating and design. AB - AIMS: This prospective, randomized study sought to directly compare the performance of paclitaxel and rapamycin on an otherwise identical, polymer-coated drug-eluting stent (DES) platform. METHODS AND RESULTS: Stents with identical design and biodegradable polymeric coating that elute either rapamycin or paclitaxel over a 2 months time period were utilized. In this pilot trial that included 91 patients, both stent platforms proved safe with no case of death, Q wave myocardial infarction or stent thrombosis within a 9 months follow-up period. Late-lumen loss was markedly greater in the paclitaxel-eluting stent group compared with the rapamycin-eluting stent group (0.96 +/- 0.75 vs. 0.33 +/- 0.46 mm, P < 0.0001). Likewise, the rate of angiographic restenosis was higher in the paclitaxel-eluting stent group compared with the rapamycin-eluting stent group [39.0 vs. 12.2%; relative risk (RR) 3.20 (95% confidence interval, 1.29 7.92), P = 0.005]. Concomitantly, the need for target lesion revascularization was higher in the paclitaxel-eluting stent group compared with the rapamycin eluting stent group [26.7 vs. 8.7%; RR 3.07 (1.07-8.80), P = 0.02]. CONCLUSION: The results of this clinical trial that is the first to directly compare the performance of paclitaxel and rapamycin on a DES platform otherwise identical in design and polymeric coating imply that rapamycin is more effective for the prevention of coronary restenosis on a DES platform with mid-term drug release and less dependent on release kinetics than paclitaxel. Thus, to ensure efficacy, drug release from a paclitaxel-coating stent platform must be prolonged and well controlled to achieve results that are comparable with the FDA-approved paclitaxel-eluting stent platform. PMID- 17921532 TI - The repertoire of desaturases and elongases reveals fatty acid variations in 56 eukaryotic genomes. AB - The repertoire of biosynthetic enzymes found in an organism is an important clue for elucidating the chemical structural variations of various compounds. In the case of fatty acids, it is essential to examine key enzymes that are desaturases and elongases, whose combination determine the range of fatty acid structures. We systematically investigated 56 eukaryotic genomes to obtain 275 desaturase and 265 elongase homologs. Phylogenetic and motif analysis indicated that the desaturases consisted of four functionally distinct subfamilies and the elongases consisted of two subfamilies. From the combination of the subfamilies, we then predicted the ability to synthesize six types of fatty acids. Consequently, we found that the ranges of synthesizable fatty acids were often different even between closely related organisms. The reason is that, as well as diverging into subfamilies, the enzymes have functionally diverged within the individual subfamilies. Finally, we discuss how the adaptation to individual environments and the ability to synthesize specific metabolites provides some explanation for the diversity of enzyme functions. This study provides an example of a potent strategy to bridge the gap from genomic knowledge to chemical knowledge. PMID- 17921534 TI - [Comparison between bare metal stents and drug eluting stents for the treatment of chronic total occlusion]. AB - Patients with chronic total occlusions (CTO) are under high risk of restenosis, repeat recanalization procedures and mortality after stent. In our study we evaluated the clinical and angiographic efficacy of bare metal stents (BMS) and drug eluting stents (DES) implantation for chronic total occlusion.107 patients were divided into two groups: who were successfully treated with BMS (n=60) 56,1% and those who had CTO lesions with Cypher implantation(n=47) 43,9%. Six-month angiographic restenosis rates and major adverse cardiac events (MACE) were compared between the groups. At 6-month angiographic follow up, the restenosis rate was significantly higher in the BMS group (21,8% vs. 4,7%; p<0,001), but there was no significant statistic difference in the rate of MACE (11,6% vs.13,3%; p>0,05). Therefore, the implantation of DES in the treatment of CTO lesions showed more favourable results regarding restenosis compared with BMS implantation, but there was not a trend towards lower MACE rates. PMID- 17921533 TI - [Relieving intra-truncal suture of tendon anastomosis]. AB - To improve the functional capacity of regenerated tendon new type of intra truncal tendon anastomosis was worked out, which was termed as "Relieving intra truncal suture". The method reduced to a minimum tissues cutting caused by the suture in the first and second phases of healing, which gave an opportunity to start active and passive movements early and with minimal risk. Clinical results showed that "the best" indices in the control group was distinctly lower (39%), than in main group (58%), which was to "Relieving intra-truncal suture" favor. PMID- 17921535 TI - [The fibroblast growth factor as a predictor of symptomatic change under combined pharmacological treatment of benign prostatic hyperplasia]. AB - The results of our study showed, that long-term combined medical treatment of benign prostatic hyperplasia (Finasteride 5 mg/day + Tamsulosin 0.4 mg/day) significantly decreases values of IPSS and residual urine volume (RUV) in patients, which basic level of bFGF in serum does not exceed 5.9 pg/ml. On the other hand pharmacological treatment was insufficient in patients with bFGF basic level equal or more than 5.9 pg/ml; values of IPSS and RUV did not decrease. PMID- 17921536 TI - Suppressive therapy after surgical treatment of thyroid gland cancer in patients with mastopathy. AB - State of lactation system during oncological disorders of thyroid gland is one of the poorly studied topics of modern medicine. Controversy in clinical and experimental data creates the need of performing studies in this direction. The goal of this study is to investigate determining pathogenic mechanisms of thyroid and lactation systems. Realization of the study results will enable development of guidelines for prophylaxis, diagnosis and treatment of mammary gland cancer in patients with thyroid gland malignancy. For this purpose we investigated patients with different types of malignant cancer of thyroid gland (after surgery), who had diffuse fibrocystic mastopathy as a concomitant disorder. Patients underwent conservative therapy only with thyroid agents. We observed patients for 6 months since the beginning of the treatment. During this period patients' condition was evaluated several times using examinations involved in the study. Summarization of study results demonstrated that suppressive therapy might prevent precancerous diseases of mammary gland. PMID- 17921537 TI - Thyroid function status and plasma lipids among cardiology patients in Georgia. AB - Thyroid dysfunction as an important cardiovascular risk factor, is not well characterized among cardiology patients of Georgia. Further, a consensus has not been reached about the relationships between thyroid function markers and plasma lipids. We investigated these risk factors among 250 cardiology patients admitted to the Emergency Cardiology Center. A cross sectional study was conducted using in-person interviews, medical records, physical exams and laboratory studies. Thyroid stimulating hormone, free thyroxine 3, free thyroxine 4 and plasma lipids were measured using standardized assays. Overall, thyroid dysfunction was detected among 28.6% of the study population (19.5% males and 39.6% females). Overt hypo- and hyperthyroidism were present among 12.4% and 6.0% of patients, while, subclinical hypo- and hyperthyroidism were present among 2.8% and 6.4% of patients respectively. Both clinical and subclinical hypothyroidism were associated with elevated total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) concentrations (p-values for trend <0.005). Further, TC and LDL-C were highly correlated with thyroid function markers (all p-values <0.000). Triglycerides and high-density lipoprotein cholesterol concentrations were not associated with thyroid function status. Hyperthyroidism was not associated with plasma lipid variation. IN CONCLUSION: thyroid dysfunction was prevalent among cardiology patients in Georgia. Hypothyroidism was associated with elevated TC and LDL-C concentrations. Future studies that examine the clinical relevance of observed differences in lipid profiles among this population are needed. PMID- 17921538 TI - [Side effects of the combined antiviral therapy in patients with chronic hepatitis C]. AB - The aim of the study is to reveal and treat the side effects of the combined antiviral therapy - Peginterferon and Ribavirin in patients with chronic Hepatitis C. During 2004-2006 years, 40 patients in the ambulatory department of the Infectious Diseases Clinic of the State Medical University were investigated. All the patients were administered pegilated interferon and ribavirin after special procedures. The following side effects on the antiviral treatment were grouped: flu-like, cardiovascular, gastrointestinal, neuro-psychiatric, dermatological and hematological. Frequency, severity and duration of all unfavorable symptoms were independent of Hepatitis C genotype, though the adherence to the medications was better in younger age groups and the dose correction where required after the age 45. PMID- 17921539 TI - [Tiotropium bromide (Spiriva) in stablechronic obstructive pulmonary disease]. AB - The present review of literature, dedicated to the usage of anticholinergic drug Tiotropium Bromide (Spiriva) in the treatment of stable chronic obstructive pulmonary disease (COPD), demonstrates its supremacy over Ipratropium Bromide (Atrovent) and sympathomimetics in relation to broncholytic effect as well as in the capacity to improve the clinical symptoms of the disease, the quality of life for the patients, prevent the exacerbations, reduce the frequency of hospitalization of the diseased in the stationer and raise the physical capacity of work. It is also important that due to its antycholinergic activity during 48 hours and of high 24-hour continuous action it is possible to prescribe Spiriva once in 24 hours. Such qualities of a drug greatly simplify regime of the dosage and improve the compliance of the patients towards the therapy. The present review gives us the possibility to discuss the effectiveness of the Tiotropium Bromide (Spiriva) as well as of its safety that puts this medicine above all other drugs used in the treatment of COPD. PMID- 17921540 TI - [Effectiveness of PCR analysis in the diagnostics of tuberculosis]. AB - To discover DNA of mycobacterium and evaluate chemotherapy efficiency of pulmonary tuberculosis Polimerize Chain Reaction (PCR) analysis was performed. Different pathological materials of patients with pulmonary tuberculosis and those with nonspecific lung diseases had been examined. Phlegm sample had been carried out regularly by means of PCR analysis and traditional microbiological methods. PCR analysis demonstrated 94% sensibility, specificity 100%; investigation of different pathological materials demonstrated 54% sensibility of molecular - genetic method. Discontinuance of bacterium secretion at PCR analysis positively correlated with clinical rontgenologic dynamics of disappearance of tubercular process. PMID- 17921541 TI - Vertigo syndromes associated with earthquake in Georgia. AB - Georgia experienced strong earthquake, after this event the amount of patients with vestibular symptoms increased. Study evaluates 60 outpatients (51 females, 9 males) aged from 18 to 85 years old who applied at outpatient clinics with vestibular complaints in the initial weeks following the earthquake. BPPV and PPV are the most common causes of vertigo. "Idiopathic" BPPV is the cause of BPPV in 50-70% of cases. Head trauma, vestibular neuritis, Meniere disease, migraine are the most common causes of "secondary" BPPV. (PPV) syndrome characterized by dizziness, subjective disturbance of balance and by perception of illusory body perturbations, usually triggered by perceptual stimulus. PPV frequently associated with anxiety symptoms in patients with obsessive-compulsive type personality. The study revealed "Idiopathic" BPPV in 49 cases and "secondary" types BPPV in 11 cases. 37 previously documented histories of BPPV patients had features typical for posterior semisercircular canal BPPV and 23 patients experienced subjective disturbance symptoms like BPPV, but there were no abnormal responses in their vestibular testing. It is assumed that earthquake could manifest psychogenic vertigo with panic attacks, anxiety, agoraphobia, PPV and could provoke a transition from organic vertigo to PPV. It is concluded that psychological stress play an important role in occurring "secondary" BPPV, earthquake may trigger exacerbation of "secondary" BPPV and could be provoked factor to developing psychogenic vertigo, mostly PPV. PMID- 17921542 TI - Effectiveness of canalith repositioning manoeuvers (CRM) in patients with benign paroxysmal positional vertigo (BPPV). AB - BPPV is the most common disorder of the vestibular system affected up to 21% of vertigo patients, characterised by short-lasting episodes of vertigo in association with rapid changes in head position. We have detected, treated and followed up 204 patients with BPPV during two years. 204 patients (163 women 41men) were enrolled in this study, at the time of evolution the duration of symptoms varied from several days to several months. We evaluated idiophatic forms of BPPV in 126 cases and secondary types BPPV in 78 cases. All patients with typical history of BPPV underwent neurological examination, including Dix Hallpike (to detect posterior and anterior canal BPPV) and Roll Test (to detect horizontal canal BPPV). We treat patients with CRM and followed up them in 48 hours and 7 days, one month, 6 month and one year after initial management. Posterior semicircular canal was involved in 170 patients, remain 34 patients suffered from canalolithiasis of horisontal (31 patients) and anterior (3 patients) semicircular canal. After a single treatment session the symptoms disappeared completely in 139 patients, in 40 patients after twice, 16 patients after 3 times and 9 patients after 4 times maneuver sessions. No effectiveness was found in 5 patients; during follow-up 7 successfully treated patients experienced a recurrence between 1 and 6 months, in two cases after one year; they were retreated and all achieved a positive result. It is concluded that diagnosis of BPPV is simple and save, do not need expensive neuroradiological tests; CRM provides effective and long term control of symptoms in patients with BPPV. PMID- 17921543 TI - [Functional state of serotoninergic system on a background of different pharmacokinetic of Midazolam]. AB - A comprehensive investigation of the high-performance liquid chromatographic separation of Midazolam and his ozidized metabolits, their bases and other low molecular-weight UV-absorbing compounds that might be found in serum is reported. The drug was administrated (2,0mg/kg) to 40 white mouses and the blood samples were collecetd for further analysis followind 30, 60, 90, 120 and 180 min. after the injection. The serotoninergic systems has been analyzed. Quantitative data were calculated by inner standart method. Oxidazed forms of Midazolam were qualitatively and quantitatively identified. Our data proved the ability of Midazolam to adsorb on the albumin and globulin fraction of plasma. Metabolism of serotonin and triptopchan is closely dependent on the concentration of psychotropic drug. Concentration of aminoacids (triptophan, tirosyn) in the blood is synshronized with the course of pharmacokinetic of the psychotropic preparation and functional state of serotoninergic system. Alterations may be due, on the one hand, to exhaustion of the stores, and on the other hand, to deterioration of the monoamines' synthesis process. The investigation of pharmacokinetics of invariable form of Midazolam and its oxidized metabolits is very important for practical medicine. Effective action of pshychotropic drugs is impossible without extensive knowlege of their phramacokinetics and pharmacodynamics. They are concerned with the need to dermine concentration in body fluids. PMID- 17921544 TI - Alterations of some macro- and microelements concentration in the rat hippocampus following destruction of its inputs during learning and memory. AB - Alterations of Na, K, Ca, and Mg concentration in the rat hippocampus were investigated with atomic-absorption method, in the process of learning and memory, following destruction of the medial septum and entorhinal cortex. These data were compared to those of the heavy metals, obtained in similar conditions. It was found that in learning and memory processes, after ablation of the main inputs of the hippocampus, sodium and potassium concentration remains relatively unaltered, while Mg and Ca undergo significant changes. In our opinion this fact is due to both increased compensatory synthesis processes induced by ablation of the main inputs of the hippocampus, and to participation of these elements in synthesis of specific proteins in a course of learning and memory and formation and retention of an engram. Increased concentration of zinc and copper certifies for important part of these metals during the above processes. PMID- 17921545 TI - [Variability of radiation-induced adaptive response in old age individuals and their correction by Peptide bioregulator -Livagen]. AB - Effect of aging on adaptive response of cellular systems to low (stimulated) dozes of gamma-rays (0, 2 and 0, 5 Gy) and to disturbing dozes of radiation (1 and 2 Gy) has been investigated. PHA-stimulated cells were from 72-86 year-old individuals; control - 30-40 year-old individuals. The potentialities of induction of adaptive response in cells exposed to previously irradiated by stimulating dozes of gamma-rays with subsequent damaging effect of copper chloride (10(-3)M) has been investigated. The correcting activity of the peptide bioregulator Livagen was tested. The investigation showed that cells from aged individuals maintained radiation adaptive feature. Preliminary exposure to radiation caused stimulation of adaptive response in copper-treated cells. Corrective activity of Livagen was observed. PMID- 17921546 TI - Study of rat's emotional state in "conflict" situations. AB - The trials performed in the rats revealed that the creation of conflict situations, in the presence of two different motivations simultaneously, results in increase in anxiety state in animals. In terms of evolving and stabilizing the two active avoidance responses in animals, the three cohorts of different learning abilities (good, average and poor) were identified in parental, as well as in filial F1 and F2 generations. While influencing with stress-factors the percentage of rats having a good learning ability decreased, while the number of rats having a poor learning ability increased, respectively. The rising of emotional background was observed through the generations, which confirms that the creation of conflict situations, which affects only one third of the parental individuals as a stress factor, becomes a stress-agent for more and more rats in the further generations. Development of pathologic conditions in animals develops from the anxiety disorder, which further intensifies through the next generations. PMID- 17921547 TI - [Basic hemodynamic and vegetative regulatory indices in adolescents-healthy and sick with arterial hypertension]. AB - Basic hemodynamic indices in aged 15-16 practically healthy adolescents do not differ essentially, but in patients with juvenile arterial hypertension is noticed their authentic rise in proper age groups. The correlation of body mass is marked with the basic hemodynamic parameters and likewise the indices of double production depend on systolic pressure and heart rate. In adolescents troubled with juvenile arterial hypertension is noticed intensification of sympathetic link of regulatory mechanisms of vegetative nervous system and unsatisfactory hypersympathetic reaction on orthostatic test which shows the total disbalance of the organism. The latter can be considered as one of the pathologic aspects of development of juvenile arterial hypertension; on the other hand non-invasitive investigation of functional state of vegetative nervous system maybe recommended for inclusion in the project of preventive medical practice to early revealing juvenile arterial hypertension or its predisposition to it. PMID- 17921548 TI - [Content and metabilism of nitric oxide in substrates of oral cavity and their role in pathogenesis of periodontite disease]. AB - Content and metabolism of free NO in saliva, fluids of periodontal pockets and gingival tissue has been investigated by EPR (electron paramagnetic resonance) method with the use of nitric oxide spin-trap in patients suffering from periodontitis. It was found that increased generation of nitric oxide at early stages of periodontitis is of protective nature. However, at serious disorders of mitochondrial respiration and exaggerated formation of generators of reactive oxygen, nitric oxide converts into citotoxic peroxinitrite leading to destruction of parodontal tissue; part of free nitric oxide produces FeSNO, which in turn, supports and decreases content of free NO in gingival tissue. As a result, decreases local antimicrobial protection, hemocirculation, tissue nutrition, progresses inflammation. Periodontitis decreases local immunity due to NO deficiency in gingival tissue. Reduced content of NO in gingival tissue decreases regeneration ability of cells, which in turn, leads to deterioration on gingival tissue. PMID- 17921549 TI - [Influense of stimulation of Substancia Nigra and Nucleus Caudatus on postsynaptic responses of cortical nociceptive neurons]. AB - The aim of the research was to investigate the influence of electric stimulation of Substancia Nigra and Nucleus Caudatus on postsynaptic processes, evoked in the nociceptive neuron of somatosensory cortex of cats. Intracellular recordings of electrical activity of the somatosensory cortex neurons in response to stimulation of the tooth pulp and of ventro-postero-medial (VPM) nucleus of thalamus, revealed the EPSP-peak-IPSP complex. Conditioning stimulation of Substancia Nigra or Nucleus Caudatus, which preceded the test-stimulation of the tooth pulp or VPM, with intervals of 100 to 700 ms, induced 40-50% decrease of IPSP only, in both nociceptive and convergent neurons. The maximal effect in both cases was achieved at an interval of 400-700 ms between conditioning- and test stimuli. Probably these processes are realized through either pre- or postsynaptic mechanisms. PMID- 17921550 TI - Biochemical structure of Calendula officinalis. AB - Calendula officinalis is a well known medicinal herb. It is common knowledge that its medicinal properties are conditioned on biologically active complex substances of Carotin (Provitamin A), Stearin, Triterpiniod, Plavonoid, Kumarin, macro and micro compound elements. Because of constant need in raw material of Calendula officinalis, features of its ontogenetic development agro-biological qualities in various eco regions of Georgia were investigated. The data of biologically active compounds, biochemical structure and the maintenance both in flowers and in others parts of plant is presented; the pharmacological activity and importance in medicine was reviewed. PMID- 17921551 TI - [Use of Plaferon LB for cardiac preconditioning during experimental ischemia/reperfusion injury in rabbits]. AB - The main goal of research was to study potential of Plaferon LB for cardiac preconditioning during experimental ischemia/reperfusion injury in rabbits. 30 rabbits (2.5-3.0 kg) were used in experiment. They were divided in 3 groups and 6 subgroups (n=5). In I group experimental design of m/i was performed by proximal ligation of left coronary artery (LCA) (2-6 hours). In II group on the 2 and 6 hour ligature was removed - reperfusion during 1 hour. In III group before ligation of LCA animals was administered Plaferon LB (0.2 mg/kg). The animals were under electrocardiographic monitoring. Troponin I was measured in blood. In II group after 1 hour of reperfusion Troponin I concentration was higher than in I group after 2 and 6 hours. In II group electrocardiographic data was worsened (rhythm and heart rate). In III group these changes were less marked. Obtained data confirm enhancement of myocardial injury during the reperfusion. Cardiac preconditioning by Plaferon LB significantly decreased pathologic indices. PMID- 17921552 TI - Prevention of thalassemia in India. PMID- 17921553 TI - Role of dexamethasone and oral glycerol in reducing hearing and neurological sequelae in children with bacterial meningitis. AB - OBJECTIVE: To investigate the efficacy of dexamethasone and oral glycerol in reducing hearing and neurological sequelae in children with acute bacterial meningitis (ABM). DESIGN: Prospective double blind, placebo controlled randomized study. SETTING: Pediatric services of a tertiary care teaching and referral hospital. SUBJECTS: Children 2 months to 12 years with a diagnosis of acute bacterial meningitis admitted between June 2002 to September 2003. INTERVENTION: Subjects were assigned randomly to receive dexamethasone, glycerol, dexamethasone+glycerol or placebo. Neurological and hearing impairment was assessed at discharge and after 1 month. RESULTS: 58 children (48 boys, 10 girls), mean age 50.2 +/- 41.0 months, were studied. Twelve patients received dexamethasone, 13 glycerol, 20 dexamethasone + glycerol and 13 placebo. Bacterial etiology was ascertained in 24 patients: Streptococcus pneumoniae-10, H influenzae b-7, Staph. aureus-5 and others-2. Three (5.2%) children died during hospital stay and 55 survived. Seven (12%) patients had neurological sequelae (3 in glycerol, 3 in dexamethasone+glycerol, 1 in placebo group, P = 0.29), and 10 patients (17%) had hearing sequelae (2 in glycerol, 3 in dexamethasone, 2 dexamethasone + glycerol and 3 in placebo group, P = 0.68). CONCLUSION: No significant difference was seen in neurological or hearing outcome with use of either glycerol or dexamethasone in children with acute bacterial meningitis. PMID- 17921554 TI - Metabolic syndrome in childhood obesity. AB - OBJECTIVES: We determined the frequency of metabolic risk factors and the prevalence of Metabolic Syndrome in childhood obesity. SUBJECTS: 186 obese children (97 females and 89 males), aged 11.2 +/- 2.8 (6-16) years and 98 healthy children (46 females and 52 males), aged 10.9 +/- 3.2 (6-16) years were recruited for the study, as study and control groups, respectively. METHODS: Subjects were evaluated for anthropometry, blood pressure (BP) and biochemical cardiovascular risk factors. Metabolic syndrome was defined in presence of > or = 3 of the following: (i) fasting triglyceride > or = 100 mg/dL; (ii) high density lipoprotein-cholesterol < 50 mg/dL, except in boys aged 15 to 19 years, in whom the cut-off point was 45 mg/dL; (iii) fasting glucose > or = 110 mg/dL; (iv) waist circumference > 75th percentile for age and gender and (v) systolic BP > 90th percentile. RESULTS: We found that 144 (77.4%) children in the obese group had one, two or more cardiovascular risk factors. Using a pediatric definition, the prevalence of metabolic syndrome was 2.1%. In the control group, the clustering of one, two and three risk factors was very rare. CONCLUSION: Childhood obesity is associated with increased frequency of cardiovascular risk factors and metabolic syndrome. PMID- 17921555 TI - Sytematic review and meta-analysis of prevalence of hepatitis B in India. AB - OBJECTIVE: To evaluate the point prevalence of Hepatitis B in India. DESIGN: Meta analysis of data on point prevalence from different parts of the country. DATA SOURCES: Searches were made in Medline, Cochrane Library and Best bets and previous reviews. A limited hand search of cross references was also done. Finally a consultation with experts was held to enlarge the references base. REVIEW METHODS: Studies reporting prevalence of HBsAg were selected. Data from high risk groups were excluded. MAIN RESULTS: 54 papers reporting data on 61 populations were identified. The true prevalence for each study was calculated from the reported prevalence using the specificity and sensitivity of the test employed. The true prevalence in non-tribal populations is 2.4% (95% CI: 2.2% 2.7%). True prevalence among tribal populations is 15.9% (CI: 11.4%-20.4%). CONCLUSION: These figures may be useful in estimation of the burden of the disease in the country and for projecting the cost-benefits of immunization. PMID- 17921557 TI - Validation of the PELOD score for multiple organ dysfunction in children. AB - We conducted a prospective study to identify the children having multiple organ dysfunction at admission using the PELOD score, and its impact on the mortality in a pediatric intensive care unit of a tertiary care hospital in north India over a 13 month period. Data were collected in a predesigned collection sheet and the PELOD score was calculated. 209 patients were admitted. In 37.2% primary indication of admission was severe sepsis/ septic shock. Ninety-one percentage of children admitted had multiple organ dysfunction. The area under the curve for predicting death using PELOD score equation was 0.80. PMID- 17921556 TI - Impact of training on observer variation in chest radiographs of children with severe pneumonia. AB - BACKGROUND: Pneumonia diagnosed using chest radiographs is often used as a study end point in trials and epidemiological studies. We studied whether training of the end-users in 172 standardized chest radiographic features will decrease variability in the interpretation. METHODS: Inter-observer variation of 3 observers in recognizing standardized radiographic features for pneumonia was studied in 172 chest radiographs of children with clinical severe pneumonia. (as per WHO definition). The observers were then trained using a software with a repository of normal and abnormal films showing a spectrum of radiological changes in pneumonia. The interobserver variation in recognizing the same standardized radiographic features was recorded after this training. For each radiographic feature, Cohen's kappa statistics to assess the between-observer agreement and Fleiss's multiple rater kappa statistics to assess agreement among all three clinicians was used. RESULTS: The 'uniterpretable' films reduced from 16.6% (95% CI 0%-34.1%) before training to 8.1% (95% CI 0%-17.7%) after training. The 'adequate' films increased from 54.2% (95% CI 12.5%-95.9%) before training to 70% (95% CI 46.5%-93.4%) after training. For all features, agreement between observers 1 with 2 and 1 with 3, the Cohen's kappa improved from poor to moderate agreement. The Fleiss's kappa values before training were 0.1 to 0.2 and after training ranged from 0.37 to 0.52 indicating moderate to good agreement after training. CONCLUSIONS: Training of the doctors using standardized features with the help of a software improves agreement substantially in identifying radiological pneumonia. PMID- 17921558 TI - Role of leptin in growth and adiposity in early infancy: impact of nutritional pattern. AB - This study was planned to investigate leptin levels under different models of nutrition and their relation with anthropometric parameters in early infancy. Sixty four infants (28 exclusively breast-fed, 15 exclusively formula-fed and 21 mixed-fed), aged 4 months were included and their leptin, insulin-like growth factor-I (IGF-I), and IGF binding protein-3 (IGFBP-3) levels were measured. Leptin, IGF-I and IGFBP-3 levels of the three groups were not statistically different (P > 0.05). The results of our study questions the role of leptin as a link between nutritional pattern and adiposity-growth in early infancy. PMID- 17921559 TI - PALS update 2005. AB - Many of the changes in BLS recommended in 2005 Guidelines are designed to simplify CPR recommendations, increase the number and quality of chest compressions delivered, and increase the number of uninterrupted chest compressions. The recommendations for compressions have been summarized as, "Push harder, push faster, allow the chest to fully recoil, and stop only to use a bag mask to ventilate the patient, analyze the rhythm, deliver a shock or intubate. When such an interruption to compressions occurs, keep the length of that interruption to an absolute minimum. For lay rescuers, a single compression ventilation ratio (30:2) for all age groups greatly simplifies the instructions for performing CPR. Recommendation of 1 Shock plus Immediate CPR for Attempted Defibrillation for cardiac arrest associated with VF or pulseless VT. Rescuers should not interrupt chest compressions to check circulation until about 5 cycles or approximately 2 minutes of CPR have been provided after the shock. The changes are designed to minimize interruptions in chest compressions. For Neonatal resuscitation, additional evidence was available about the use of oxygen versus room air for resuscitation, the need for clearing the airway of meconium, methods of assisting ventilation, techniques for confirming endotracheal tube placement, and use of the laryngeal mask airway (LMA). PMID- 17921560 TI - Gianotti Crosti syndrome. PMID- 17921562 TI - Incessant atrial flutter after device closure of atrial septal defect: successful radio frequency ablation. AB - A four-month-old baby presented with failure to thrive and congestive cardiac failure precipitated by a lower respiratory tract infection. He was found to have a large ostium secundum atrial septal defect measuring 18 mm. This was successfully closed percutaneously by a device (Blockaid). A month after the device deployment the child developed typical atrial flutter. Despite rate control drugs the ventricular rate remained 140/min over the next several months. In view of the incessant atrial flutter with fast ventricular response, the child underwent radiofrequency ablation at the age of 2 years. An isthmus block was created which successfully terminated the tachycardia. PMID- 17921561 TI - Epididymoorchitis and pancytopenia caused by brucellosis. AB - A 15 year old boy presented with fever and acute painful scrotal swelling. Complete blood count showed pancytopenia. Serum brucella antibodies were positive. Pancytopenia and epididymoorchitis are rare complications of brucellosis and clinicians must consider this entity in the differential diagnosis of adolescents with epididiymoorchitis associated with pancytopenia. PMID- 17921563 TI - Rabies encephalitis. AB - A 12-year-old boy presented with fever and sore throat of 6 days duration followed by vomiting and altered sensorium. He had received 4 doses of antirabies vaccine following a dog bite 4 weeks back. Rabies immunoglobulin was not given. History of hydrophobia and aerophobia were strikingly absent. The possibilities of rabies encephalitis and vaccine induced acute disseminated encephalomyelitis (ADEM) were considered. MRI brain showed exclusive grey matter changes characteristic of rabies. The diagnosis was further confirmed by serological tests. PMID- 17921564 TI - National immunisation coverage: need for surveillance. PMID- 17921565 TI - Leptin resistance in obese Indian girls? PMID- 17921566 TI - Media responsibility and child health. PMID- 17921568 TI - IPV--doubts persist. PMID- 17921570 TI - First in-beam PET measurement of beta+ radioactivity induced by hard photon beams. AB - In this note, we present the first experimental results of in-beam PET measurements during high energy photon phantom irradiation. An inhomogeneous phantom was irradiated with pulsed 34 MV bremsstrahlung. The measurements have been conducted with a dedicated double head positron camera. A high material contrast could be achieved and furthermore production rates of (11)C and (15)O were derived from the time-dependent activity. PMID- 17921571 TI - Acoustical properties of selected tissue phantom materials for ultrasound imaging. AB - This note summarizes the characterization of the acoustic properties of four materials intended for the development of tissue, and especially breast tissue, phantoms for the use in photoacoustic and ultrasound imaging. The materials are agar, silicone, polyvinyl alcohol gel (PVA) and polyacrylamide gel (PAA). The acoustical properties, i.e., the speed of sound, impedance and acoustic attenuation, are determined by transmission measurements of sound waves at room temperature under controlled conditions. Although the materials are tested for application such as photoacoustic phantoms, we focus here on the acoustic properties, while the optical properties will be discussed elsewhere. To obtain the acoustic attenuation in a frequency range from 4 MHz to 14 MHz, two ultrasound sources of 5 MHz and 10 MHz core frequencies are used. For preparation, each sample is cast into blocks of three different thicknesses. Agar, PVA and PAA show similar acoustic properties as water. Within silicone polymer, a significantly lower speed of sound and higher acoustical attenuation than in water and human tissue were found. All materials can be cast into arbitrary shapes and are suitable for tissue-mimicking phantoms. Due to its lower speed of sound, silicone is generally less suitable than the other presented materials. PMID- 17921572 TI - Megavoltage x-ray skin dose variation with an angle using grid carbon fibre couch tops. AB - It is well known that a skin dose from high-energy x-ray radiation varies with the angle of beam incidence or the presence of a radiotherapy linear accelerator couch top material. This note investigates changes produced to the skin dose from a Varian carbon fibre grid couch top at differing angles of incidence for 6 MV x rays as is often the case clinically. Results have shown that the skin dose can easily be measured using an EBT Gafchromic film whereby the delivered skin dose can be quantified to a high level of spatial resolution, not easily achieved with other skin dose detectors. Results have shown a significant increase in the skin dose specifically at the point of a cross-sectional carbon fibre grid. Values in % of the skin dose increased from approximately 27% (an open area within a 10 cm x 10 cm field) up to 55% (same field size) at the centre of the carbon fibre mesh strip (0 degrees incidence). This is compared to 19% of the skin dose for an open field of a 10 cm x 10 cm beam without the couch material present. At larger angles similar effects occur with values changing from 52% to 75% (60 degrees , 10 cm x 10 cm) in the open area and under the grid, respectively. This produces a wave effect for the skin dose. The average skin dose magnitude increases with the angle of incidence of the beam, ranging from 37.5% to 66% from 0 degrees to 60 degrees (10 x 10 cm), respectively. The symmetric wave nature of the skin dose profile skews to deliver an increased dose on the posterior side of the carbon fibre grid as the angle of incidence increases. Simulated fractional dose delivery on a phantom has shown that over 30 fractions the wave nature of the delivered skin dose is minimized due to the random nature of most patient positioning on the treatment couch. However, some variations are still present as the ratio of the open to grid area is approximately 4:1 and the dose spread is not necessarily completely averaged during a typical fractionated radiotherapy treatment. As such, if the treatment type results in a more rigorously positioned patient on the treatment couch, the wave nature of skin dose delivery may need to be taken into account. PMID- 17921573 TI - Comparison of intensity modulated x-ray therapy and intensity modulated proton therapy for selective subvolume boosting: a phantom study. AB - Selective subvolume boosting can theoretically improve tumour control probability while maintaining normal tissue complication probabilities similar to those of uniform dose distributions. In this work the abilities of intensity-modulated x ray therapy (IMXT) and intensity-modulated proton therapy (IMPT) to deliver boosts to multiple subvolumes of varying size and proximities are compared in a thorough phantom study. IMXT plans were created using the step-and-shoot (IMXT SAS) and helical tomotherapy (IMXT-HT) methods. IMPT plans were created with the spot scanning (IMPT-SS) and distal gradient tracking (IMPT-DGT) methods. IMPT-DGT is a generalization of the distal edge tracking method designed to reduce the number of proton beam spots required to deliver non-uniform dose distributions relative to IMPT-SS. The IMPT methods were delivered over both 180 degrees and 360 degrees arcs. The IMXT-SAS and IMPT-SS methods optimally satisfied the non uniform dose prescriptions the least and the most, respectively. The IMPT delivery methods reduced the normal tissue integral dose by a factor of about 2 relative to the IMXT delivery methods, regardless of the delivery arc. The IMPT DGT method reduced the number of proton beam spots by a factor of about 3 relative to the IMPT-SS method. PMID- 17921574 TI - A large-scale study of the ultrawideband microwave dielectric properties of normal, benign and malignant breast tissues obtained from cancer surgeries. AB - The development of microwave breast cancer detection and treatment techniques has been driven by reports of substantial contrast in the dielectric properties of malignant and normal breast tissues. However, definitive knowledge of the dielectric properties of normal and diseased breast tissues at microwave frequencies has been limited by gaps and discrepancies across previously published studies. To address these issues, we conducted a large-scale study to experimentally determine the ultrawideband microwave dielectric properties of a variety of normal, malignant and benign breast tissues, measured from 0.5 to 20 GHz using a precision open-ended coaxial probe. Previously, we reported the dielectric properties of normal breast tissue samples obtained from reduction surgeries. Here, we report the dielectric properties of normal (adipose, glandular and fibroconnective), malignant (invasive and non-invasive ductal and lobular carcinomas) and benign (fibroadenomas and cysts) breast tissue samples obtained from cancer surgeries. We fit a one-pole Cole-Cole model to the complex permittivity data set of each characterized sample. Our analyses show that the contrast in the microwave-frequency dielectric properties between malignant and normal adipose-dominated tissues in the breast is considerable, as large as 10:1, while the contrast in the microwave-frequency dielectric properties between malignant and normal glandular/fibroconnective tissues in the breast is no more than about 10%. PMID- 17921575 TI - Neutron stimulated emission computed tomography: a Monte Carlo simulation approach. AB - A Monte Carlo simulation has been developed for neutron stimulated emission computed tomography (NSECT) using the GEANT4 toolkit. NSECT is a new approach to biomedical imaging that allows spectral analysis of the elements present within the sample. In NSECT, a beam of high-energy neutrons interrogates a sample and the nuclei in the sample are stimulated to an excited state by inelastic scattering of the neutrons. The characteristic gammas emitted by the excited nuclei are captured in a spectrometer to form multi-energy spectra. Currently, a tomographic image is formed using a collimated neutron beam to define the line integral paths for the tomographic projections. These projection data are reconstructed to form a representation of the distribution of individual elements in the sample. To facilitate the development of this technique, a Monte Carlo simulation model has been constructed from the GEANT4 toolkit. This simulation includes modeling of the neutron beam source and collimation, the samples, the neutron interactions within the samples, the emission of characteristic gammas, and the detection of these gammas in a Germanium crystal. In addition, the model allows the absorbed radiation dose to be calculated for internal components of the sample. NSECT presents challenges not typically addressed in Monte Carlo modeling of high-energy physics applications. In order to address issues critical to the clinical development of NSECT, this paper will describe the GEANT4 simulation environment and three separate simulations performed to accomplish three specific aims. First, comparison of a simulation to a tomographic experiment will verify the accuracy of both the gamma energy spectra produced and the positioning of the beam relative to the sample. Second, parametric analysis of simulations performed with different user-defined variables will determine the best way to effectively model low energy neutrons in tissue, which is a concern with the high hydrogen content in biological tissue. Third, determination of the energy absorbed in tissue during neutron interrogation in order to estimate the dose. Results from these three simulation experiments demonstrate that GEANT4 is an effective simulation platform that can be used to facilitate the future development and optimization of NSECT. PMID- 17921577 TI - Correlation of energy dispersive diffraction signatures and microCT of small breast tissue samples with pathological analysis. AB - Identification of specific tissue types in conventional mammographic examinations is extremely limited. However, the use of x-ray diffraction effects during imaging has the potential to characterize the tissue types present due to the fact that each tissue type produces its own unique diffraction signature. Nevertheless, the analysis and categorization of these diffraction signatures by tissue type can be hampered by the inhomogeneous nature of breast tissue, leading to categorization errors where several types are present. This work aims to reduce sample categorization errors by combining spectral diffraction signature collection with sample imaging, giving more detailed data on the composition of each sample. Diffraction microCT was carried out on 19 unfixed breast tissue samples using an energy resolving translate-rotate CT system. High-resolution transmission microCT images were also recorded for comparison and sample composition analysis. Following imaging, the samples were subjected to histopathological analysis. Reconstructing on various momentum transfer regions allows different tissue types to be identified in the diffraction images. Results show a correlation between measured x-ray diffraction images and stained histopathological tissue sections. X-ray diffraction signatures generated from the measured data were categorized and analysed, with a t-test indicating that they have the potential for use in tissue type identification. PMID- 17921576 TI - Statistical image reconstruction from correlated data with applications to PET. AB - Most statistical reconstruction methods for emission tomography are designed for data modeled as conditionally independent Poisson variates. In reality, due to scanner detectors, electronics and data processing, correlations are introduced into the data resulting in dependent variates. In general, these correlations are ignored because they are difficult to measure and lead to computationally challenging statistical reconstruction algorithms. This work addresses the second concern, seeking to simplify the reconstruction of correlated data and provide a more precise image estimate than the conventional independent methods. In general, correlated variates have a large non-diagonal covariance matrix that is computationally challenging to use as a weighting term in a reconstruction algorithm. This work proposes two methods to simplify the use of a non-diagonal covariance matrix as the weighting term by (a) limiting the number of dimensions in which the correlations are modeled and (b) adopting flexible, yet computationally tractable, models for correlation structure. We apply and test these methods with simple simulated PET data and data processed with the Fourier rebinning algorithm which include the one-dimensional correlations in the axial direction and the two-dimensional correlations in the transaxial directions. The methods are incorporated into a penalized weighted least-squares 2D reconstruction and compared with a conventional maximum a posteriori approach. PMID- 17921578 TI - A Fourier reconstruction algorithm with constant attenuation compensation using 180 degrees acquisition data for SPECT. AB - In this paper, we develop an approximate analytical reconstruction algorithm that compensates for uniform attenuation in 2D parallel-beam SPECT with a 180-degree acquisition. This new algorithm is in the form of a direct Fourier reconstruction. The complex variable central slice theorem is used to derive this algorithm. The image is reconstructed with the following steps: first, the attenuated projection data acquired over 180 degrees are extended to 360 degrees and the value for the uniform attenuator is changed to a negative value. The Fourier transform (FT) of the image in polar coordinates is obtained from the Fourier transform of an analytic function interpolated from an extension of the projection data according to the complex central slice theorem. Finally, the image is obtained by performing a 2D inverse Fourier transform. Computer simulations and comparison studies with a 360-degree full-scan algorithm are provided. PMID- 17921579 TI - Dynamic contrast-enhanced CT of head and neck tumors: perfusion measurements using a distributed-parameter tracer kinetic model. Initial results and comparison with deconvolution-based analysis. AB - The objective of this work was to evaluate the feasibility of a two-compartment distributed-parameter (DP) tracer kinetic model to generate functional images of several physiologic parameters from dynamic contrast-enhanced CT data obtained of patients with extracranial head and neck tumors and to compare the DP functional images to those obtained by deconvolution-based DCE-CT data analysis. We performed post-processing of DCE-CT studies, obtained from 15 patients with benign and malignant head and neck cancer. We introduced a DP model of the impulse residue function for a capillary-tissue exchange unit, which accounts for the processes of convective transport and capillary-tissue exchange. The calculated parametric maps represented blood flow (F), intravascular blood volume (v(1)), extravascular extracellular blood volume (v(2)), vascular transit time (t(1)), permeability-surface area product (PS), transfer ratios k(12) and k(21), and the fraction of extracted tracer (E). Based on the same regions of interest (ROI) analysis, we calculated the tumor blood flow (BF), blood volume (BV) and mean transit time (MTT) by using a modified deconvolution-based analysis taking into account the extravasation of the contrast agent for PS imaging. We compared the corresponding values by using Bland-Altman plot analysis. We outlined 73 ROIs including tumor sites, lymph nodes and normal tissue. The Bland-Altman plot analysis revealed that the two methods showed an accepted degree of agreement for blood flow, and, thus, can be used interchangeably for measuring this parameter. Slightly worse agreement was observed between v(1) in the DP model and BV but even here the two tracer kinetic analyses can be used interchangeably. Under consideration of whether both techniques may be used interchangeably was the case of t(1) and MTT, as well as for measurements of the PS values. The application of the proposed DP model is feasible in the clinical routine and it can be used interchangeably for measuring blood flow and vascular volume with the commercially available reference standard of the deconvolution-based approach. The lack of substantial agreement between the measurements of vascular transit time and permeability-surface area product may be attributed to the different tracer kinetic principles employed by both models and the detailed capillary tissue exchange physiological modeling of the DP technique. PMID- 17921580 TI - Dosimetric characteristics of electron beams produced by a mobile accelerator for IORT. AB - Energy and angular distributions of electron beams with different energies were simulated by Monte Carlo calculations. These beams were generated by the NOVAC7 system (Hitesys, Italy), a mobile electron accelerator specifically dedicated to intra-operative radiation therapy (IORT). The electron beam simulations were verified by comparing the measured dose distributions with the corresponding calculated distributions. As expected, a considerable difference was observed in the energy and angular distributions between the IORT beams studied in the present work and the electron beams produced by conventional accelerators for non IORT applications. It was also found that significant differences exist between the IORT beams used in this work and other IORT beams with different collimation systems. For example, the contribution from the scattered electrons to the total dose was found to be up to 15% higher in the NOVAC7 beams. The water-to-air stopping power ratios of the IORT beams used in this work were calculated on the basis of the beam energy distributions obtained by the Monte Carlo simulations. These calculated stopping power ratios, s(w,air), were compared with the corresponding s(w,air) values recommended by the TRS-381 and TRS-398 IAEA dosimetry protocols in order to estimate the deviations between a dosimetry based on generic parameters and a dosimetry based on parameters specifically obtained for the actual IORT beams. The deviations in the s(w,air) values were found to be as large as up to about 1%. Therefore, we recommend that a preliminary analysis should always be made when dealing with IORT beams in order to assess to what extent the possible differences in the s(w,air) values have to be accounted for or may be neglected on the basis of the specific accuracy needed in clinical dosimetry. PMID- 17921581 TI - Comparison of Newtonian and non-Newtonian flows in a two-dimensional carotid artery model using the lattice Boltzmann method. AB - Numerical modelling is a powerful tool for the investigation of human blood flow and arterial diseases such as atherosclerosis. It is known that near wall shear is important in the pathogenesis and progression of atherosclerosis. When modelling arterial blood flow it is generally assumed that blood is Newtonian. In this paper, blood flow is modelled in a realistic two-dimensional carotid artery geometry in order to investigate this assumption and its effect on the measurement of near wall shear. The assumption is tested in stenosed and unstenosed geometries and the non-Newtonian blood is modelled using the Carreau Yasuda model. It is found that the velocity and shear fields, particularly near the walls of the geometries, exhibit small differences in general (<5%) between Newtonian and non-Newtonian models, even in the stenosed geometry with peak differences of 13.6%. Thus, when using numerical modelling to study the haemodynamic influences on atherosclerotic progression, we can safely neglect the non-Newtonian nature of blood. PMID- 17921582 TI - Multiple template-based fluoroscopic tracking of lung tumor mass without implanted fiducial markers. AB - Precise lung tumor localization in real time is particularly important for some motion management techniques, such as respiratory gating or beam tracking with a dynamic multi-leaf collimator, due to the reduced clinical tumor volume (CTV) to planning target volume (PTV) margin and/or the escalated dose. There might be large uncertainties in deriving tumor position from external respiratory surrogates. While tracking implanted fiducial markers has sufficient accuracy, this procedure may not be widely accepted due to the risk of pneumothorax. Previously, we have developed a technique to generate gating signals from fluoroscopic images without implanted fiducial markers using a template matching method (Berbeco et al 2005 Phys. Med. Biol. 50 4481-90, Cui et al 2007 Phys. Med. Biol. 52 741-55). In this paper, we present an extension of this method to multiple-template matching for directly tracking the lung tumor mass in fluoroscopy video. The basic idea is as follows: (i) during the patient setup session, a pair of orthogonal fluoroscopic image sequences are taken and processed off-line to generate a set of reference templates that correspond to different breathing phases and tumor positions; (ii) during treatment delivery, fluoroscopic images are continuously acquired and processed; (iii) the similarity between each reference template and the processed incoming image is calculated; (iv) the tumor position in the incoming image is then estimated by combining the tumor centroid coordinates in reference templates with proper weights based on the measured similarities. With different handling of image processing and similarity calculation, two such multiple-template tracking techniques have been developed: one based on motion-enhanced templates and Pearson's correlation score while the other based on eigen templates and mean-squared error. The developed techniques have been tested on six sequences of fluoroscopic images from six lung cancer patients against the reference tumor positions manually determined by a radiation oncologist. The tumor centroid coordinates automatically detected using both methods agree well with the manually marked reference locations. The eigenspace tracking method performs slightly better than the motion-enhanced method, with average localization errors less than 2 pixels (1 mm) and the error at a 95% confidence level of about 2-4 pixels (1-2 mm). This work demonstrates the feasibility of direct tracking of a lung tumor mass in fluoroscopic images without implanted fiducial markers using multiple reference templates. PMID- 17921583 TI - Calorimetric determination of kQ factors for NE 2561 and NE 2571 ionization chambers in 5 cm x 5 cm and 10 cm x 10 cm radiotherapy beams of 8 MV and 16 MV photons. AB - The relative uncertainty of the ionometric determination of the absorbed dose to water, D(w), in the reference dosimetry of high-energy photon beams is in the order of 1.5% and is dominated by the uncertainty of the calculated chamber- and energy-dependent correction factors k(Q). In the present investigation, k(Q) values were determined experimentally in 5 cm x 5 cm and 10 cm x 10 cm radiotherapy beams of 8 MV and 16 MV bremsstrahlung by means of a water calorimeter operated at 4 degrees C. Ionization chambers of the types NE 2561 and NE 2571 were calibrated directly in the water phantom of the calorimeter. The measurements were carried out at the linear accelerator of the Physikalisch Technische Bundesanstalt. It is shown that the k(Q) factor of a single ionization chamber can be measured with a standard uncertainty of less than 0.3%. No significant variations of k(Q) were found for the different lateral sizes of the radiation fields used in this investigation. PMID- 17921584 TI - Regulation of cerebral endothelial cell morphology by extracellular calcium. AB - Cerebral endothelial cells interconnected by tight and adherens junctions constitute the structural basis of the blood-brain barrier. Extracellular calcium ions have been reported to play an important role in the formation and maintenance of the junctional complex. However, little is known about the action of calcium depletion on the structural characteristics of cerebral endothelial cells. Using atomic force microscopy we analyzed the effect of calcium depletion and readdition on the shape and size of living brain endothelial cells. It was found that the removal of extracellular calcium from confluent cell cultures induced the dissociation of the cells from each other accompanied by an increase in their height. After readdition of calcium a gradual recovery was observed until total confluency was regained. We have also demonstrated that Rho-kinase plays an important role in the calcium-depletion-induced disassembly of endothelial tight and adherens junctions. The Rho-kinase inhibitor Y27632 could prevent the morphological changes induced by a lack of calcium as well. Our results suggest that calcium depletion induces Rho-kinase-dependent cytoskeletal changes that may be partly responsible for the disassembly of the junctional complex. PMID- 17921585 TI - Efficiency of antiscatter grids for flat-detector CT. AB - Flat-panel detector CT (FD-CT) scanners offer large volume coverage, but as a consequence are more susceptible to scatter artifacts than standard clinical CT scanners with smaller cone angles. FD-CT scanners can employ antiscatter grids as a scatter rejection technique. We evaluated three standard fluoroscopic antiscatter grids for two different field sizes with respect to scatter suppression efficiency and image quality improvement. The evaluations included simulations and measurements. Regarding the simulation a hybrid model combining deterministic and Monte Carlo (MC) calculations was used combined with an analytical calculation of grid transmission. The scatter-to-primary ratio (SPR) was measured using an adapted collimator technique in order to validate our simulations. The SPR obtained by simulations and measurements with and without antiscatter grids were in agreement typically within 10%. The employment of a grid does not generally provide a significant improvement of the signal-to-noise ratio (SNR). Antiscatter grids led to a significant reduction of cupping artifacts in all cases. There is a trade-off between the SNR and the reduction of the scatter intensity described by the signal-to-noise improvement factor (SNR(if)). For low- or medium-scatter conditions the increase in noise caused by the reduced primary transmission through the grid has to be compensated by a higher exposure. For high scatter conditions SNR(if) is significantly greater than 1; i.e. a decrease of dose of up to 50% can be reached. PMID- 17921586 TI - Variation of haemoglobin extinction coefficients can cause errors in the determination of haemoglobin concentration measured by near-infrared spectroscopy. AB - Near-infrared spectroscopy or imaging has been extensively applied to various biomedical applications since it can detect the concentrations of oxyhaemoglobin (HbO(2)), deoxyhaemoglobin (Hb) and total haemoglobin (Hb(total)) from deep tissues. To quantify concentrations of these haemoglobin derivatives, the extinction coefficient values of HbO(2) and Hb have to be employed. However, it was not well recognized among researchers that small differences in extinction coefficients could cause significant errors in quantifying the concentrations of haemoglobin derivatives. In this study, we derived equations to estimate errors of haemoglobin derivatives caused by the variation of haemoglobin extinction coefficients. To prove our error analysis, we performed experiments using liquid tissue phantoms containing 1% Intralipid in a phosphate-buffered saline solution. The gas intervention of pure oxygen was given in the solution to examine the oxygenation changes in the phantom, and 3 mL of human blood was added twice to show the changes in [Hb(total)]. The error calculation has shown that even a small variation (0.01 cm(-1) mM(-1)) in extinction coefficients can produce appreciable relative errors in quantification of Delta[HbO(2)], Delta[Hb] and Delta[Hb(total)]. We have also observed that the error of Delta[Hb(total)] is not always larger than those of Delta[HbO(2)] and Delta[Hb]. This study concludes that we need to be aware of any variation in haemoglobin extinction coefficients, which could result from changes in temperature, and to utilize corresponding animal's haemoglobin extinction coefficients for the animal experiments, in order to obtain more accurate values of Delta[HbO(2)], Delta[Hb] and Delta[Hb(total)] from in vivo tissue measurements. PMID- 17921587 TI - A stochastic approach for automatic registration and fusion of left atrial electroanatomic maps with 3D CT anatomical images. AB - The integration of electroanatomic maps with highly resolved computed tomography cardiac images plays an important role in the successful planning of the ablation procedure of arrhythmias. In this paper, we present and validate a fully automated strategy for the registration and fusion of sparse, atrial endocardial electroanatomic maps (CARTO maps) with detailed left atrial (LA) anatomical reconstructions segmented from a pre-procedural MDCT scan. Registration is accomplished by a parameterized geometric transformation of the CARTO points and by a stochastic search of the best parameter set which minimizes the misalignment between transformed CARTO points and the LA surface. The subsequent fusion of electrophysiological information on the registered CT atrium is obtained through radial basis function interpolation. The algorithm is validated by simulation and by real data from 14 patients referred to CT imaging prior to the ablation procedure. Results are presented, which show the validity of the algorithmic scheme as well as the accuracy and reproducibility of the integration process. The obtained results encourage the application of the integration method in post intervention ablation assessment and basic AF research and suggest the development for real-time applications in catheter guiding during ablation intervention. PMID- 17921588 TI - A novel approach to multi-criteria inverse planning for IMRT. AB - Treatment plan optimization is a multi-criteria process. Optimizing solely on one objective or on a sum of a priori weighted objectives may result in inferior treatment plans. Manually adjusting weights or constraints in a trial and error procedure is time consuming. In this paper we introduce a novel multi-criteria optimization approach to automatically optimize treatment constraints (dose volume and maximum-dose). The algorithm tries to meet these constraints as well as possible, but in the case of conflicts it relaxes lower priority constraints so that higher priority constraints can be met. Afterwards, all constraints are tightened, starting with the highest priority constraints. Applied constraint priority lists can be used as class solutions for patients with similar tumour types. The presented algorithm does iteratively apply an underlying algorithm for beam profile optimization, based on a quadratic objective function with voxel dependent importance factors. These voxel-dependent importance factors are automatically adjusted to reduce dose-volume and maximum-dose constraint violations. PMID- 17921589 TI - Biologically-equivalent dose and long-term survival time in radiation treatments. AB - Within the linear-quadratic model the biologically-effective dose (BED)-taken to represent treatments with an equal tumor control probability (TCP)-is commonly (and plausibly) calculated according to BED(D) = -log[S(D)]/alpha. We ask whether in the presence of cellular proliferation this claim is justified and examine, as a related question, the extent to which BED approximates an isoeffective dose (IED) defined, more sensibly, in terms of an equal long-term survival probability, rather than TCP. We derive, under the assumption that cellular birth and death rates are time homogeneous, exact equations for the isoeffective dose, IED. As well, we give a rigorous definition of effective long-term survival time, T(eff). By using several sets of radiobiological parameters, we illustrate potential differences between BED and IED on the one hand and, on the other, between T(eff) calculated as suggested here or by an earlier recipe. In summary: (a) the equations currently in use for calculating the effective treatment time may underestimate the isoeffective dose and should be avoided. The same is the case for the tumor control probability (TCP), only more so; (b) for permanent implants BED may be a poor substitute for IED; (c) for a fractionated treatment schedule, interpreting the observed probability of cure in terms of a TCP formalism that refers to the end of the treatment (rather than T(eff)) may result in a miscalculation (underestimation) of the initial number of clonogens. PMID- 17921590 TI - 40 years of JCO interviews. PMID- 17921592 TI - Effects of missed appointments and bracket failures on treatment efficiency and office productivity. PMID- 17921591 TI - Fact or friction: the clinical relevance of in vitro steady-state friction studies. PMID- 17921593 TI - The readers' corner. Efficient tooth movement methods and cosmetic finishing. PMID- 17921595 TI - The rationale for maxillary second premolar extractions in adult Class II treatment. PMID- 17921596 TI - Rapid tooth movement with a low-force, low-friction bracket system. PMID- 17921598 TI - Dr. Eugene L. Gottlieb on 40 years of JCO. Interview by Robert G. Keim. PMID- 17921597 TI - Digital hand-wrist radiographs for evaluating skeletal maturation. PMID- 17921599 TI - The future of orthodontics: part 1 diagnosis and treatment. PMID- 17921600 TI - Complex orthodontic treatment using a new protocol for the Invisalign appliance. PMID- 17921601 TI - Orthodontics in the year 2047: genetically driven treatment plans. PMID- 17921602 TI - The evolution of digital study models. PMID- 17921603 TI - Integrating space closure and esthetic dentistry in patients with missing maxillary lateral incisors. PMID- 17921604 TI - Pigmented contact dermatitis. PMID- 17921606 TI - Art and science of patch testing. PMID- 17921607 TI - Thin-layer rapid-use epicutaneous test (TRUE test). PMID- 17921608 TI - Parthenium: a wide angle view. PMID- 17921609 TI - Relationship between nickel allergy and diet. AB - Nickel is a ubiquitous trace element and it occurs in soil, water, air and of the biosphere. It is mostly used to manufacture stainless steel. Nickel is the commonest cause of metal allergy. Nickel allergy is a chronic and recurring skin problem; females are affected more commonly than males. Nickel allergy may develop at any age. Once developed, it tends to persist life-long. Nickel is present in most of the dietary items and food is considered to be a major source of nickel exposure for the general population. Nickel content in food may vary considerably from place to place due to the difference in nickel content of the soil. However, certain foods are routinely high in nickel content. Nickel in the diet of a nickel-sensitive person can provoke dermatitis. Careful selection of food with relatively low nickel concentration can bring a reduction in the total dietary intake of nickel per day. This can influence the outcome of the disease and can benefit the nickel sensitive patient. PMID- 17921610 TI - Patch testing experience with 1000 patients. AB - BACKGROUND: Patch testing is a definitive tool for diagnosing allergic contact dermatitis (ACD). It reveals the prevalence and trends of contact sensitization in the community, thereby paving the way for better standard series. There is paucity of large series of patch-tested patients from India. AIM: To report the 9 year patch-test data from a single general dermatology centre in North India. METHODS: Consecutive patients presenting with signs/symptoms of suspected ACD were patch tested from May 1997 to April 2006. The Indian Standard Series was used. Parthenium was tested only in selected patients and cetrimide and chloroxylenol were added to the series. RESULTS: In total, records of 1000 patients (566 male, 434 female) were analyzed, yielding 1155 positive reactions in 590 (59%) patients. Footwear dermatitis was the commonest suspected diagnosis, followed by ACD to medicaments, cosmetic dermatitis and plant dermatitis. Out of the allergens that were tested in all the patients, positivity to nickel was the commonest (12.9%), followed by potassium dichromate (11.1%) neomycin (7%), mercaptobenzthiazole (6.6%), nitrofurazone (6%), colophony (5.7%), fragrance mix (5.5%) and cobalt chloride (5.4%). However, parthenium was the commonest allergen based on the proportion of patients tested with it (14.5%). In men, potassium dichromate (30%) was the commonest sensitizer and in women, nickel (43%) was the commonest to show patch-test positivity. CONCLUSION: Our study revealed higher prevalence of footwear and medicament dermatitis in comparison to existing data. Allergy to antiseptics is significant in our patients. Further collaborative studies involving patients from other parts of India are required to have an overall view of ACD in India. PMID- 17921611 TI - Epidemio-allergological study in 155 cases of footwear dermatitis. AB - BACKGROUND: Footwear dermatitis represents a distinct and common group among all types of contact dermatitis cases seen in India. This ailment, however, often remains undiagnosed, misdiagnosed or empirically diagnosed without pinpointing the contributory allergens. AIMS: This study was undertaken to detect the epidemio-allergological pattern of footwear dermatitis in India. METHODS: A total number of 155 cases with footwear dermatitis were evaluated from July 2005 to June 2006, by detailed history and clinical examination. They were patch tested using Indian Standard Battery (ISB) approved by the Contact and Occupational Dermatoses Forum of India (CODFI) with pre- and post patch-test counseling. RESULTS: The proportion of footwear dermatitis was 24.22% (n=155) among a total of 640 patients patch tested during that period. Females [61.93% (n=96)] were commonly affected than males [38.06% (n=59)]. The ages ranged from 8 to 75 years. The age group that predominantly involved was the fifth decade [24.52% (n=38)]. Occupationwise housewives were most commonly involved [47.48 (n=66)]. Contributory allergens in order of frequency were: potassium dichromate, 45.8% (n=71); cobalt chloride, 38.06% (n=59); paraphenylenediamine, 32.25% (n=50); epoxy resin, 20% (n=31); black rubber mix, 20% (n=31); nickel sulfate, 14.83% (n=23); mercaptobenzothiazole, 12.9% (n=20); colophony, 11.6% (n=18); thiuram mix, 10.32 % (n=16); p-tert-butyl-formaldehyde resin, 9.67% (n=15); and formaldehyde, 4.5% (n=7). Among the different categories of footwear allergens, the highest positivity was shown by leather and leather-related chemicals in 61.9% cases (n=96). CONCLUSION: Footwear dermatitis, a common dermatosis, is mostly caused by leather processing chemicals, metal buckles, black dyes of shoes and socks, adhesives, plastic, rubber shoes and polishing agents in order of frequency. PMID- 17921612 TI - Periorbital melanosis is an extension of pigmentary demarcation line-F on face. AB - BACKGROUND: To date, periorbital melanosis is an ill-defined entity. The condition has been stated to be darkening of the skin around the eyes, dark circles, infraorbital darkening and so on. AIMS: This study was aimed at exploring the nature of pigmentation in periorbital melanosis. METHODS: One hundred consecutive patients of periorbital melanosis were examined and investigated to define periorbital melanosis. Extent of periorbital melanosis was determined by clinical examination. Wood's lamp examination was performed in all the patients to determine the depth of pigmentation. A 2-mm punch biopsy was carried out in 17 of 100 patients. RESULTS: In 92 (92%) patients periorbital melanosis was an extension of pigmentary demarcation line over the face (PDL-F). CONCLUSION: Periorbital melanosis and pigmentary demarcation line of the face are not two different conditions; rather they are two different manifestations of the same disease. PMID- 17921613 TI - Adapalene pretreatment increases follicular penetration of clindamycin: in vitro and in vivo studies. AB - BACKGROUND: Topical retinoids normalize desquamation, reduce comedogenesis and may enhance the penetration of other topicals providing more effective treatment of acne. AIM: We evaluated the effect of adapalene on skin penetration of clindamycin phosphate when it is applied concomitantly or after various time durations following adapalene application. METHODS: The in vitro studies were carried out using excised rat skin, whereas the in vivo studies were conducted on healthy human volunteers. Radioactive clindamycin phosphate (1%) gel was applied to rat skin sections and to the hands of human volunteers concomitantly and after the pretreatment of the skin for 3, 5 and 10 min with 10 mg of adapalene (0.1%) gel. Quantification of clindamycin phosphate was performed by liquid scintillation. RESULTS: In vitro skin penetration and distribution of clindamycin phosphate was affected by the pretreatment time. Significantly higher skin concentration of clindamycin phosphate (15.5%) with largest proportion in viable skin layer (9.4% of applied dose) was found when clindamycin phosphate gel was applied after the pretreatment of the skin with adapalene gel for 5 min. Further increase in pretreatment time has no additive influence on the penetration of clindamycin phosphate. In vivo results were in corroboration with the in vitro results and demonstrate significantly higher concentration of clindamycin phosphate (19%) in the skin following pretreatment with adapalene gel for 5 min. Adapalene acts as a penetration enhancer and increases the penetration of topical clindamycin phosphate. CONCLUSION: Application of clindamycin phosphate gel after the pretreatment of skin with adapalene gel for 5 min may contribute significantly to the increased efficacy of therapy. PMID- 17921614 TI - Cutaneous reactions simulating erythema multiforme and Stevens Johnson syndrome due to occupational exposure to a plant-growth regulator. AB - BACKGROUND: In India, hydrogen cyanamide (Dormex) is a plant growth regulator used mainly for the bud-breaking of grapevines. The use of this chemical may result in severe cutaneous reactions simulating erythema multiforme (EM), Stevens Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN). METHODS: Studies were conducted on four seasonal grapevine workers who developed severe cutaneous reactions following the unprotected use of Dormex (hydrogen cyanamide). RESULTS: Two of the patients had EM-like skin lesions and the other two developed SJS-TEN like skin lesions. A latent period of 5-7 days existed between the contact with the chemical and the development of the skin lesions. The histopathological picture was suggestive of EM. All the patients responded to systemic steroids and antihistamines. CONCLUSIONS: Hydrogen cyanamide may act as a hapten, initiating cytotoxic immunological attack on keratinocytes, resulting in EM- and SJS-TEN like clinical picture. Awareness regarding such severe cutaneous reactions due to the inappropriate handling of Dormex is required. The use of personal protection equipment while handling agricultural chemicals is essential. PMID- 17921615 TI - Clinicoepidemiological profile of 590 cases of beetle dermatitis in western Orissa. AB - BACKGROUND: Beetle dermatitis is a very common condition in western Orissa. It is often misdiagnosed and causes significant morbidity among the rural population. AIM: This study was conducted to determine the epidemiological and clinical profile of beetle dermatitis in western Orissa. METHODS: All clinically diagnosed cases of beetle dermatitis were included in the study. Detailed history was taken and thorough clinical examination was conducted in all the cases. One urban and three rural localities were visited regularly to detect the epidemiological trends of the disorder. RESULTS: A total of 590 cases were studied: 486 males and 104 females. The age of the patients ranged from 2 to 65 years. Forty-four percent of the patients belonged to the pediatric age group. Majority of the cases (85%) presented during the months of March to July, indicating a distinct seasonal trend. The disorder was prevalent in the localities nearer to paddy and sugarcane fields and grasslands with stagnant water. The clinical lesions included papules, erosions, crusted lesions, urticarial plaques and vesiculobullous lesions. Distribution was mainly linear, but kissing lesions were also observed. Head, neck and upper extremities were the most commonly involved sites. Lymphadenopathy and systemic features such as fever and malaise were observed in 24% and 15% of the cases, respectively. CONCLUSION: Beetle dermatitis should be included in the differential diagnosis of the acute onset of vesiculobullous lesions in the endemic areas. PMID- 17921617 TI - Congenital erythropoietic porphyria in three siblings. AB - Congenital erythropoietic porphyria is a rare autosomal recessive disorder that usually presents with marked skin photosensitivity, hypertrichosis, blistering, scarring, milia formation and dyspigmentation of the photo-exposed areas. Three adult siblings (two sisters and one brother) are presented here with variable degree of skin manifestations. During early childhood, all the siblings started showing signs of photosensitivity with darkening of urine color followed by skin blistering over the face and hands. The oldest showed severe sclerodermiform mutilation and the youngest exhibited an initial involvement with hypertrichosis. None of them had any history of convulsions, acute abdominal pain or joint pain. Woods lamp examination and laboratory investigations confirmed the diagnosis. PMID- 17921616 TI - 32P-patch contact brachyradiotherapy in the management of recalcitrant keloids and hypertrophic scars. AB - Keloids are the result of excessive fibroblast proliferation and then over abundant collagen deposition. There is no method able to guarantee absolute success in the therapeutic approach to keloids. Our case report involves a female patient with six lesions treated with a 32P-patch brachyradiotherapy. Pre treatment and adjuvant treatment of the lesions were performed with thiomucase, 5 fluoruracil, procaine and triamcinolone. Taking into account the activity contained in each of the patches and the total radiation dose to be administered according to clinical practice, dosimetric calculations were done for each lesion. Separate silicone patches with chromic [32P]phosphate were designed for each lesion based on these calculations. Total remission was achieved in three treated lesions. The other lesions did not achieve total remission yet, but their sizes are diminishing. The differences observed in treatment outcome may be related with lesion features, adjuvant treatments and/or treatment schedule. PMID- 17921618 TI - Disseminated cutaneous rhinosporidiomas in an immunocompetent male. AB - Rhinosporidiosis is a chronic recurrent infective granulomatous disease of man and animals. It is endemic in India and Sri Lanka. Rhinosporidiosis is a chronic disease commonly involving the nose and nasopharynx. Cutaneous lesions, although rare, can occur due to autoinoculation or due to hematogenous spread. However, disseminated cutaneous lesions presenting as tumor-like swellings are rare. We report here a 48-year-old immunocompetent patient who had disseminated painless cutaneous tumor-like swellings over both the upper limbs, abdomen, left buttock and calf since 10 months, gradually increasing in size. On inquiring, the patient gave history of excisions and electrocauterization of subglottic and nasal polyps. Histopathological examination of these lesions was suggestive of rhinosporidiosis. The general and systemic examinations of the patient did not reveal any abnormality. PMID- 17921619 TI - Angiolymphoid hyperplasia with follicular mucinosis. AB - Follicular mucinosis occurring along with angiolymphoid hyperplasia with eosinophils (ALHE) has been described in a 54-year-old female. The patient presented with pruritic erythematous papules on the left frontoparietal scalp. Histopathological examination showed prominent blood vessels in the dermis lined by plump histiocytoid endothelial cells that were surrounded by a dense lymphoid infiltrate with numerous eosinophils; these findings are typical of angiolymphoid hyperplasia with eosinophils. Features of follicular mucinosis were observed in the same section with several hyperplastic follicular infundibula containing pools of mucin in the infundibular epithelium. The concurrent occurrence of these two distinct histopathological patterns in the same biopsy specimen has been described in only three cases to date. PMID- 17921620 TI - Mycophenolate mofetil as adjuvant in pemphigus vulgaris. AB - Pemphigus vulgaris (PV) is a life threatening autoimmune blistering disease of skin and mucous membranes. Advent of systemic steroids has greatly reduced the mortality rate. However, steroids and adjuvant immunosuppressive therapy are nowadays frequent contributory agents of morbidity and mortality of PV. Mycophenolate mofetil (MMF) has been reported to be an effective adjuvant to systemic steroids. It helps in increasing the immunosuppressive effect and minimizing the toxicities by steroid sparing effect. However, its efficacy in refractory cases of PV is not well documented. The lowest possible dose with satisfactory therapeutic efficacy and least side effects is known. We used MMF 1 g/day and systemic steroids in 3 Indian patients with pemphigus vulgaris who were resistant to systemic steroid monotherapy or combination treatment with azathioprine. In our experience, MMF offers an effective adjuvant with minimal side-effects in the treatment of resistant PV. PMID- 17921621 TI - Determination of minimum erythema dose for narrow band UVB therapy and skin typing. PMID- 17921622 TI - Persistent hiccups: a rare prodromal manifestation of herpes zoster. PMID- 17921623 TI - An adult with a cord round the neck: benign transient lymphangiectasis of the penis. PMID- 17921624 TI - Human immunodeficiency virus and pemphigus vulgaris: an interesting association. PMID- 17921625 TI - Hereditary leukonychia totalis. PMID- 17921626 TI - Subcutaneous fat necrosis of the newborn mimicking generalized lymphadenopathy. PMID- 17921627 TI - Clear cell hidradenoma in a young boy. PMID- 17921628 TI - Acquired crateriform hyperkeratotic papules of the feet: an unusual variant of focal acral hyperkeratosis. PMID- 17921629 TI - Localized cutaneous leishmaniasis in South India: successful treatment with ketoconazole. PMID- 17921630 TI - Darier's sign. PMID- 17921631 TI - Eflornithine. PMID- 17921633 TI - Crouzons syndrome: a case report. AB - Human skull is made up of many bone joints connected by sutures. The sutures fuse in later life after the complete growth of the brain. If any of these sutures closes early, it may interfere with the normal growth of the brain. The developing brain may exert pressure on the skull and may grow in the direction of the other open sutures. Premature sutural fusion may occur alone or together with other anomalies, making up various syndromes. Crouzons syndrome is an example of such a syndrome that is associated with premature synostosis of the sutures of the skull. Presented in this article is a case of Crouzons syndrome seen in a boy aged 9 years. PMID- 17921632 TI - Multiple asymptomatic nodules in a middle-aged patient. Tophaceous gout. PMID- 17921634 TI - Management of traumatically intruded permanent incisors. AB - Intrusive luxation of permanent teeth has great psychologic impact on both parents and children and presents clinical challenge for the dentist, all the more due to severe complications it is difficult to manage this group of dental injuries. This paper describes the management of traumatically intruded permanent maxillary-central incisor in an 11-year-old girl. After 12-month follow-up, it was seen that teeth were fully re-erupted and are in functional alignment with other teeth. PMID- 17921635 TI - Delayed replantation of avulsed teeth. AB - Dental injuries are very common and their extent has been classified by Ellis. Avulsion of tooth is a grievous injury and ranges from 1-16% among the traumatic injuries, of which maxillary anterior are commonest. Reimplantation of avulsed teeth is a standard procedure. However, it has certain limitations. Most often their management is very challenging. In this case report we are presenting the management of maxillary incisors by replantation after 36 hrs in a 12 year old girl. PMID- 17921636 TI - Restoring biological width in crown-root fracture: a periodontal concern. AB - A 10-year-old male child reported to J.S.S. Dental College and Hospital presenting with a vertical crown-root fracture of maxillary central incisor with a history of fall 10 days back. A new treatment protocol was attempted in contrary to various schools of thoughts of coronal fragment removal wherein fractured fragments were approximated and internally reinforced with latest technologies in adhesive dentistry, the goal was to save the tooth, restore its function and esthetics. This case presents the above mentioned philosophy. PMID- 17921637 TI - Management of a rare combination of dental trauma: a case report. AB - The occurrence of combined injury of intrusion, avulsion and lateral luxation is rare and the mechanism responsible for this is intriguing. This case report describes such a combined injury and its management. The rationale behind the treatment modalities is discussed. PMID- 17921638 TI - Garre's sclerosing osteomyelitis. AB - Garre's sclerosing osteomyelitis is a specific type of chronic osteomyelitis that mainly affects children and young adults. This disease entity is well-described in dental literature and is commonly associated with an odontogenic infection resulting from dental caries. This paper describes a case of Garre's osteomyelitis in a 10-year-old boy, in whom the condition arose following pulpoperiapical infection in relation to permanent mandibular right first molar. Clinically the patient presented with bony hard, non-tender swelling and the occlusal radiograph revealed pathognomic feature of "onion skin" appearance. The elimination of periapical infection was achieved by endodontic therapy and the complete bone remodeling was seen radiographically after three months follow-up. PMID- 17921639 TI - Treatment of severely mutilated incisors: a challenge to the pedodontist. AB - Dental caries is the single most common chronic childhood disease. In early childhood caries, there is early carious involvement and gross destruction of the maxillary anterior teeth. This leads to difficulty in speech, decreased masticatory efficiency, development of abnormal tongue habits and subsequent malocclusion and psychological problems if esthetics are compromised. The restoration of severely decayed primary incisors is often a difficult procedure that presents a special challenge to pediatric dentists. This case report documents the restoration of severely mutilated lateral incisors in a patient with early childhood caries. PMID- 17921641 TI - Syndromes... the clinician's nightmare? PMID- 17921640 TI - Glanzmann's thrombasthenia associated with HBsAg-positive child: a case report. AB - Glanzmann's thrombasthenia is a rare hemorrhagic disorder characterized by prolonged bleeding time and diminished clot retraction. The disease is marked by frequent mucocutaneous hemorrhage which is mainly due to qualitative defects of platelets. A case of a 14-year-old HBsAg-positive adolescent male with Glanzmann's thrombasthenia has been presented. PMID- 17921642 TI - Unusual neonatal tooth in maxillary 1st molar region: a case report. AB - Teeth erupting within the first month after birth are known as neonatal teeth. Incidence of neonatal teeth is very low. Neonatal teeth erupt in various regions of the maxillary and mandibular arch. Incidence of neonatal teeth in molar region is only 1%. A case of an unusual neonatal tooth in the maxillary molar region has been presented. PMID- 17921644 TI - Bird-headed dwarf of Seckel. AB - Seckel syndrome is an extremely rare inherited disorder characterized by growth delays prior to birth resulting in low birth weight. Growth delays continue after birth resulting in short stature (dwarfism). This syndrome is associated with an abnormally small head, varying degrees of mental retardation and unusual "beak like" protrusion of nose. Other facial features may include abnormally large eyes, a narrow face, malformed ears and an unusually small jaw. This syndrome has an autosomal recessive pattern of inheritance. A case of the Seckel syndrome is presented. PMID- 17921643 TI - Ellis Van Creveld syndrome. AB - Ellis Van Creveld syndrome is a rare disorder and is a form of short-limbed dwarfism. It is an autosomal recessive disorder characterized by tetrad of disproportionate dwarfism, post-axial polydactyly, ectodermal dysplasia and heart defects. This case report presents a classical case of a seven-year-old boy with Ellis Van Creveld syndrome presented with discrete clinical findings. PMID- 17921645 TI - Central nervous system mycoses: the challenges. PMID- 17921646 TI - Fungi: too evolved to be condemned. PMID- 17921647 TI - Epidemiology of central nervous system mycoses. AB - Fungal infections of the central nervous system (CNS) were considered rare until the 1970s. This is no longer true in recent years due to widespread use of corticosteroids, cytotoxic drugs and antibiotics. Immunocompromised patients with underlying malignancy organ transplantations and acquired immune deficiency syndrome are all candidates for acquiring fungal infections either in meninges or brain. A considerable number of cases of CNS fungal infections even in immunocompetent hosts have been reported. A vast array of fungi may cause infection in the CNS, but barring a few, most of them are anecdotal case reports. Cryptococcus neoformans, Candida albicans, Coccidioides immitis. Histoplasma capsulatum are common causes of fungal meningitis; Aspergillus spp, Candida spp, Zygomycetes and some of the melanized fungi are known to cause mass lesions in brain. Few fungi like C. neoformans, Cladophialophora bantiana, Exophiala dermatitidis, Ramichloridium mackenzie, Ochroconis gallopava are considered as true neurotropic fungi. Most of the fungi causing CNS infection are saprobes with worldwide distribution; a few are geographically restricted like Coccidioides immitis. The infections reach the CNS either by the hematogenous route or by direct extension from colonized sinuses or ear canal or by direct inoculation during neurosurgical procedures. PMID- 17921648 TI - Pathobiology of fungal infections of the central nervous system with special reference to the Indian scenario. AB - Ubiquitously present fungi in the environment find a nidus in the human body and adopt its metabolic machinery to be in symbiosis or become pathogenic. Immunocompromised states like human immunodeficiency virus (HIV) / acquired immunodeficiency syndrome (AIDS), systemic neoplasia and organ transplantation have enhanced the frequency of fungal infections. High-risk behavior, IV drug abuse and air travel have led to the emergence of new fungal infections hitherto geographically localized. The pathology in the central nervous system (CNS) is dictated largely by the size of the fungus - the yeast forms, by virtue of their small size enter the microcirculation to cause meningitis and microabscesses, while hyphal forms invade the vasculature to manifest as large pale or hemorrhagic infarcts. The growth kinetics of fungi, the antigenic character of the capsule. the proteases secreted by the mycelial forms and the biochemical milieu in the host also determine clinical manifestations. A hospital-based analysis of the available information from India suggests that in the non-HIV patient population, hyphal forms like Aspergillosis and Zygomycosis are the most common pathogens, while yeast forms like Cryptococcus and Candida are the prime pathogens in cases of HIV/AIDS, the altered macrophage function acting in synergy with suppressed cell-mediated immunity. In Northeastern states, systemic infection by Penicillium marneffei is reported in association with HIV though CNS involvement is not recorded. Although fungal infections of the CNS are reported from various hospitals in India, studies are limited by non-availability of relevant microbiological studies and the reported prevalence data is biased by the surgical practices, availability of postmortem and microbiology and laboratory support. Detailed clinical and mycological investigations related to the interaction between the fungus and host environment is a fertile area of research to understand the basic pathogenetic mechanisms. PMID- 17921649 TI - Immunopathogenesis of central nervous system fungal infections. AB - Fungal infections of the central nervous system (CNS) evoke humoral and cellular immune responses with the scope to enable the host to eliminate the pathogen. Immunopathogenesis of CNS fungal infections remains incompletely understood, with most of our understanding coming from studies on experimentally infected animals. However, activation of brain resident cells combined with relative expression of immunoenhancing and immunosuppressing cytokines and chemokines may play a determinant role and partially explain immunopathogenesis of CNS fungal infections. PMID- 17921650 TI - Fungal infections of the central nervous system: the clinical syndromes. AB - Fungal infections of the central nervous system (CNS) are being increasingly diagnosed both in immunocompromised and immunocompetent individuals. Sinocranial aspergillosis is more frequently described from countries with temperate climates, more often in otherwise immunocompetent individuals. The clinical syndromes with which fungal infections of the CNS can present are protean and can involve most part of the neuroaxis. Certain clinical syndromes are specific for certain fungal infections. The rhinocerebral form is the most common presenting syndrome with zygomycosis and skull-base syndromes are often the presenting clinical syndromes in patients with sinocranial aspergillosis. Subacute and chronic meningitis in patients with HIV infection is more likely to be due to cryptococcal infection. Early recognition of the clinical syndromes in an appropriate clinical setting is the first step towards achieving total cure in some of these infections. PMID- 17921651 TI - Cryptococcal meningitis: clinical, diagnostic and therapeutic overviews. AB - Cryptococcal meningitis has emerged as a leading cause of infectious morbidity and mortality in patients with AIDS. Among the human immunodeficiency virus (HIV) seropositive subjects, cryptococcal meningitis is the second most common cause of opportunistic neuro-infection. Current trends are changing due to the marked improvement of quality and length of life produced by highly active antiretroviral therapy (HAART). The introduction of generic HAART in India has resulted in an increase in the number of individuals getting treatment for HIV infection, as the cost of highly active antiretroviral therapy (HAART) has decreased 20- fold. Cryptococcal meningitis occurs in non-HIV patients who are immunodeficient due to diabetes, cancer, solid organ transplants, chemotherapeutic drugs, hematological malignancies etc and rarely in healthy individuals with no obvious predisposing factors. Diagnosis of cryptococcal meningitis is fairly straightforward once the diagnosis is considered in the differential diagnosis of chronic meningitis. Treatment of a patient with cryptococcal infection is a challenge for both the physician and the patient, but rewarding, as many would recover with timely and adequate antifungal therapy. PMID- 17921652 TI - Laboratory investigation of fungal infections of the central nervous system. AB - While fungal infections of the central nervous system (CNS) are relatively rare, fungal pathogens are increasingly being recognized as an important etiology of CNS infections, particularly amongst the growing immunocompromized population. In this paper we aim to provide a practical approach to the diagnosis of fungal infections of the CNS, review some of the diagnostic methods currently available and discuss diagnosis of certain pathogens of particular interest to the practicing neurologist. PMID- 17921654 TI - Fungal infections of the central nervous system: A review of fungal pathogens and treatment. AB - Multiple factors influence the outcome of fungal infection of the central nervous system (CNS). The host and the pathogen in concert with drug delivery across the blood-brain barrier and drug activity are key factors in outcome. Drug costs can be prohibitively expensive. Drug toxicity with standard antifungal agents such as amphotericin B (infusion rate toxicity) can be reduced using simple techniques such as slower infusion and appropriate saline loading. Continuous infusion can allow relatively large doses of amphotericin B (up to 2 mg/kg/day, remaining below 0.08 mg/kg/hour) to be given with toxicity profiles comparable to expensive lipid formulations of amphotericin B. Dedicated peripherally inserted central catheters can remain in situ for weeks to months and are safe and relatively inexpensive. Correction of metabolic pathology in the case of mucormycosis and resolution of neutropenia are essential to effective treatment of filamentous fungal infections such as Mucor, Aspergillus and Scedosporium. The pharmacology and pharmacokinetics of the current major antifungal agents used to treat fungal infections of the CNS are reviewed. Tables that provide information about achievable CNS drug levels, antifungal susceptibilities and the likelihood of intrinsic drug resistance of significant fungal pathogens have been included to help the clinician with therapy. Treatment recommendations for Cryptococcal and Candida meningitis and for rhinocerebral infection with Mucor and Aspergillus have been included. PMID- 17921653 TI - Imaging features of central nervous system fungal infections. AB - Fungal infections of the central nervous system (CNS) are rare in the general population and are invariably secondary to primary focus elsewhere, usually in the lung or intestine. Except for people with longstanding diabetes, they are most frequently encountered in immunocompromised patients such as those with acquired immunodeficiency syndrome or after organ transplantation. Due to the lack of inflammatory response, neuroradiological findings are often nonspecific and are frequently mistaken for tuberculous meningitis, pyogenic abscess or brain tumor. Intracranial fungal infections are being identified more frequently due to the increased incidence of AIDS patients, better radiological investigations, more sensitive microbiological techniques and better critical care of moribund patients. Although almost any fungus may cause encephalitis, cryptococcal meningoencephalitis is most frequently seen, followed by aspergillosis and candidiasis. The biology, epidemiology and imaging features of the common fungal infections of the CNS will be reviewed. The radiographic appearance alone is often not specific, but the combination of the appropriate clinical setting along with computed tomography or magnetic resonance may help to suggest the correct diagnosis. PMID- 17921655 TI - Immunotherapy for fungal infections with special emphasis on central nervous system infections. AB - Opportunistic fungal infections are major causes of morbidity and mortality in the immunocompromized. Fungi have evolved complex and coordinated mechanisms to survive in the environment and the mammalian host. Fungi must adapt to "stressors" in the host, including nutrient scarcity, pH and reactive oxygen and nitrogen intermediates, in addition to evading host immunity. Knowledge of the immunopathogenesis of fungal infections has paved the way to promising strategies for immunotherapy. These include strategies that increase phagocyte number, activate innate host defense pathways in phagocytes and dendritic cells and stimulate antigen-specific immunity (e.g, vaccines). Immunotherapy must be tailored to specific immunocompromized states. Our review focuses on cryptococcosis and coccidioidomycosis because of the propensity of these diseases to involve the central nervous system (CNS). The CNS has long been considered "immunologically privileged" in the sense of being isolated from normal immune surveillance. This notion is only partially accurate. Immune-based therapies for fungal CNS disease are at an exploratory level and merit further evaluation in clinical trials. PMID- 17921656 TI - Surgical management of intracranial fungal masses. AB - BACKGROUND: Intracranial fungal masses (IFMs, granulomas and abscesses) are uncommon lesions, infrequently encountered by neurosurgeons. There is no conclusive evidence on the ideal surgical management of these lesions. AIMS: To summarize the recent literature on the prevalence, presentation, surgical management and outcome of patients with IFMs. MATERIALS AND METHODS: The recent published literature was searched using standard search engines (PubMed and Google) for articles reporting on the databases and surgical management of IFMs. A special effort was made to include publications from Indian centers. RESULTS: Intracranial fungal masses were rarely seen even in major neurosurgical centers in India with a prevalence of around one to two per year. While most patients with IFM have immunosuppressed states, nearly 50% of patients with IFMs (especially in India) have no obvious predisposing causes and are apparently immunocompetent. The clinical presentation could be categorized into three groups: 1. Involvement of the cranial nerves 1 to 6 with orbital and nasal symptoms. 2. Focal neurological deficits due to involvement of any part of the neuraxis; and 3. "Stroke-like" presentation with sudden onset of hemiparesis. Based on the presence or absence of radiological evidence of paranasal sinus disease, IFMs were classified into two types: 1. Rhinocerebral type; 2. Purely intracranial type that was further divided into a. intracerebral or b. extracerebral forms. Aspergillus species was the commonest fungal organism causing IFMs but a number of other fungi have been reported to cause IFMs. Surgery for IFMs can be of different types, namely 1. Stereotactic procedures; 2. Craniotomy; 3. Shunt surgery; and 4. Treatment of fungal aneurysms. Generally, radical surgery is advocated for IFMs but there is no unanimity regarding the radicality of the excision especially for the rhinocerebral form of the disease. Surgery should always be followed by antifungal therapy for prolonged periods. Mortality and morbidity in patients with IFMs is very high and ranges from 40 92%. Immunosuppressed patients with IFMs and those in whom the diagnosis is delayed have the highest mortality rates, with immunocompetent patients with the rhinocerebral form of the disease having the best outcome. CONCLUSIONS: There should be a high index of suspicion for IFMs not only in patients with known risk factors for the development of fungal infections but also in immunocompetent patients in India. Intraoperative pathological diagnosis should be obtained in any patient suspected to have an IFM and tissue should be processed for fungal cultures. Prompt diagnosis, radical and safe surgery and aggressive and prolonged treatment with anti-fungal agents may lead to a better outcome especially in immunocompetent patients. PMID- 17921657 TI - Craniocerebral aspergillosis in immunocompetent hosts: surgical perspective. AB - Craniocerebral aspergillosis is a rare but dangerous variety of central nervous system infections. Surgery is being widely recognized as the cornerstone of management. Due to the rarity of the disease, difficulty and delay in diagnosis and poor outcome, there is very little in the literature regarding the various surgical strategies that may be adopted in these patients. Early aggressive surgery followed by chemotherapy offers the best chances. Surgical planning would depend upon the type and location of the disease process as well as the condition of the patient. Perioperative care holds immense importance and knowledge of possible complications is essential. Aspergillosis of the central nervous system is difficult to diagnose and equally difficult to treat. Surgery remains the cornerstone of management followed by systemic antifungal medications. Results are better in immunocompetent patients as compared to those who are immunocompromised. PMID- 17921658 TI - Central nervous system cladosporiosis: an account of ten culture-proven cases. AB - BACKGROUND: Central nervous system (CNS) cladosporiosis is a rare infection caused by Cladophialophora bantiana. It has varied presentation and poor outcome. Most of the available data in the literature are reviews of individual case reports. OBJECTIVE: To describe the clinical, radiological and mycological features of 10 cases of C. bantiana managed at a single tertiary center. To analyze the various treatment options, factors associated with outcome and to review the relevant literature. MATERIALS AND METHODS: This is a retrospective study of 10 patients with CNS cladosporiosis managed at National Institute of Mental Health and Neurosciences from 1979 to 2006. It is a descriptive study. The case records were reviewed for clinical presentation, radiological features, management and outcome. Only those patients in whom the fungus could be isolated on culture were included in the study. RESULTS: The age of the patients ranged from three to 42 years. Nine patients presented with features of space-occupying lesion and one patient with chronic meningitis. There were no specific clinical or radiological features. None of patients had impaired immune status. This infection presented as two pathomorphological forms - diffuse meningoencephalitis and focal abscesses. Burr hole tapping and excision are the surgical options. Both patients with burr hole tapping required excision of abscess subsequently. Two out of seven patients with abscess expired compared to all three patients with diffuse meningoencephalitis who expired. Recurrences occurred in four of the five patients following excision of the abscess. Combination antifungal treatment had better result than monotherapy. The outcome was poor with survival of only 50%. CONCLUSIONS: Thorough microbiological examination is required to diagnose CNS infection caused by C. bantiana. The outcome is better in patients with abscess. Excision of the abscess followed by combination antifungal therapy results in better outcome. Close follow-up is required due to high risk of recurrence. PMID- 17921659 TI - Isolated cerebral Aspergillus granuloma with no obvious source of infection. AB - BACKGROUND: Intracranial fungal granulomas occur by extension from contiguous structures or by hematogenous dissemination from lungs. Isolated granulomas without any obvious source of infection are extremely uncommon. OBJECTIVE: To describe isolated intracerebral Aspergillus spp. granuloma without any obvious source of infection. MATERIALS AND METHODS: We analyzed clinical, radiological and pathological features of isolated intracerebral aspergillus granulomas diagnosed in our institution between 1986 and 2006. The chest X-ray and paranasal sinus (PNS) X-rays were reviewed. Fungal stainings were done on histological sections. RESULTS: We identified eight patients with Aspergillus spp. intracerebral granulomas (six males, two females). There were no predisposing risk factors. The chest and PNS X-rays were normal. On computerized tomography all were heterogeneously enhancing lesions with perilesional edema. Pre or perioperative diagnosis was never made. Histological studies revealed granulomas with minimal fibrosis and giant cells and septate hyphae of Aspergillus spp. on fungal stains. Two patients died of postoperative complications and two patients relapsed. CONCLUSION: Isolated intracerebral aspergillus granulomas are rare and pose a diagnostic challenge. Fungal granulomas should be considered in the differential diagnosis of intracerebral inflammatory pathologies. PMID- 17921660 TI - Predictors of mortality in rhinocerebral mycosis. AB - INTRODUCTION: Rhinocerebral mycosis is a rapidly progressive fatal opportunistic infection, predominantly affecting people in an immunocompromised state. Aggressive surgical therapy, with repeated debridement in combination with intravenous amphotericin B can lead to a high rate of cure. AIM: To determine the predictors of mortality in rhinocerebral mycosis. MATERIALS AND METHODS: The demographic data, clinical features, radiological (MRI/CT) findings, treatment details of patients with a diagnosis of rhinocerebral mycosis confirmed on histopathology were analyzed retrospectively. The outcome was assessed as alive and dead. Univariate analysis with odds ratio (OR) was employed in data analysis. Chi-square test was used for P value. RESULTS: There were 38 patients. The age range was 7-82 (mean 48.68) years; 30 (79%) were males. Craniofacial pain was the most common initial presenting symptom, noted in 29 (76.3%). Rhino-orbital involvement was noted in 24 (63.2%) and 12 (31.6%) had associated focal neurological deficits. Immunocompromised state was noted in 24 (63.2%). Eighteen (47.4%) patients died. The predictors for mortality: odds ratio (95% CI) were 2.45 (1.01-3.89) for elderly age, 5.67 (4.13-7.21) for intracranial extension, 2.6 (1.26-3.94) for immunocompromised state, 2.62 (1.25-3.99) for infection with zygomycosis and 2.33 (1.01-3.65) for anemia. CONCLUSION: Rhinocerebral mycosis is associated with high mortality in spite of aggressive therapy. Intracranial extension with focal neurological deficits is a major predictor of mortality in rhinocerebral mycosis. PMID- 17921661 TI - Cerebral Aspergillus arteritis with bland infarcts: a report of two patients with poor outcome. AB - Two patients with cerebrovascular aspergillosis, in the form of arteritis, thrombosis and bland infarcts are reported. One patient had systemic lupus erythematosus with disseminated aspergillosis in lungs, kidneys and brain. The other patient was immunocompetent and had sphenoid sinusitis. Both the patients were diagnosed at autopsy only, despite extensive imaging and laboratory studies. High index of clinical suspicion and early aggressive antifungal therapy are required since definite diagnostic modalities are not available. PMID- 17921662 TI - Response of central nervous system aspergillosis to voriconazole. AB - Fungal infections of the central nervous system (CNS) usually present as subacute meningitis. Other manifestations include mass effect and focal neurological deficits. Cerebrospinal fluid (CSF) examination and biopsy of the lesion are helpful in disclosing the organism involved. Aspergillosis presents as brain abscess or granuloma with predominant neutrophils in CSF. Voriconazole is a broad spectrum triazole antifungal agent. It can be given orally and has lesser adverse effects We report a 69-years-old diabetic male, with aspergilloma of para-nasal sinus invading the CNS, who responded well to voriconazole treatment. He discontinued the medication by himself as it was costly. Within a month of stopping the medication, he developed features of subacute meningitis. However he showed clinical improvement after the medication was restarted. The case is reported for the clinical evidence of antifungal activity of voriconazole against aspergillosis. PMID- 17921663 TI - Sepsis and meningoencephalitis due to Rhodotorula glutinis in a patient with systemic lupus erythematosus, diagnosed at autopsy. AB - Rhodotorula species have been reported as a causative agent of opportunistic mycoses in immunocompromised hosts. We report a case of sepsis and meningoencephalitis caused by Rhodotorula glutinis in a 20-year-old female patient with systemic lupus erythematosus (SLE), which was diagnosed at autopsy. The patient presented with longstanding fever. She was diagnosed with SLE after admission to the hospital and died on day 5 of the hospital stay. Autopsy was performed to confirm the presence of infection. Sepsis and meningoencephalitis due to Rhodotorula glutinis was confirmed by postmortem blood cultures and histopathological examination of biopsies taken from the brain at autopsy. Infection by Rhodotorula spp. is rare but can be fatal in immunocompromised hosts. Infections by such uncommon yeasts may often be difficult to diagnose, especially in the setting of febrile neutropenia. This report also emphasizes the value of autopsy as a powerful educational tool. PMID- 17921664 TI - Disseminated aspergillosis causing spinal cord compression in a child. PMID- 17921666 TI - An isolated non-dural-based cerebellar aspergilloma in an immunocompetent patient. PMID- 17921665 TI - Invasive aspergillosis of the brain: improvement with lyposomal amphoterecin B and itraconazole. PMID- 17921667 TI - Fungal cerebellar tonsillar abscess as a cause of quadriparesis. PMID- 17921668 TI - Cryptococcal meningitis unmasking idiopathic CD4 lymphocytopenia. PMID- 17921670 TI - Endothelial function and circulating endothelial progenitor cells in patients with sleep apnea syndrome. AB - BACKGROUND: Endothelial dysfunction and cardiovascular diseases are frequent in patients with obstructive sleep apnea (OSA). Circulating endothelial progenitor cells (EPCs) contribute to repair dysfunctional endothelium and have been related to increased cardiovascular risk. OBJECTIVES: We tested the hypothesis that the number of circulating EPCs may be altered in OSA patients. METHODS: EPCs (CD34+ VEGF-R2+) were isolated and quantified from peripheral blood samples of OSA patients (n = 13) and healthy controls (n = 13) matched for age and sex. All subjects were free of any other known cardiovascular risk factors. The plasma levels of vascular endothelial growth factor (VEGF) were also determined, and the endothelium-dependent and endothelium-independent vascular function was assessed in all subjects. RESULTS: Patients with OSA had lower levels of EPCs (p < 0.05) and higher plasma levels of VEGF (p < 0.05) than controls. Endothelial function was not different between OSA and controls. CONCLUSIONS: Patients with OSA free of any other known cardiovascular risk factor show a reduced number of circulating EPCs and an increase in plasma VEGF levels. These alterations may contribute to future endothelial dysfunction in these patients. PMID- 17921671 TI - Deep caries lesions after incomplete dentine caries removal: 40-month follow-up study. AB - Radiographic changes after indirect pulp capping (32 teeth in 27 patients) were studied for up to 36-45 months. Radiolucent zone (RZ) depth and tertiary dentine formation were assessed qualitatively and changes in radiographic density (by image subtraction) in RZ and control areas (CA) were estimated. During follow-up there were 1 pulp necrosis, 1 pulp exposure, 3 fractures and 3 withdrawals. Twelve cases showed decreased RZ depth and 4 displayed tertiary dentine. No changes with time in density of CA or RZ, or in the difference between them, were observed. It is concluded that indirect pulp capping arrests lesion progression, suggesting that complete dentine caries removal is not essential for caries control. PMID- 17921672 TI - Acute stress-related gastrointestinal ischemia. AB - We report a case of acute gastrointestinal ischemia during a very stressful event in whom the diagnosis was made by 24-hour tonometry. This case report unequivocally links a stressful event with increased catecholamine release and subsequent severe symptomatic gastrointestinal ischemia. The role of ischemia as potential pathophysiological mechanism has never been studied in detail. The clinical significance of finding such an association is underscored by this case report, where a vasoactive drug normally used for hypertension treatment resulted in greatly improved abdominal symptoms. PMID- 17921673 TI - Possible pitfalls in the interpretation of microcirculatory measurements. A comparative study using intravital microscopy, spectroscopy and polarographic pO2 measurements. AB - BACKGROUND: Microcirculation and tissue oxygenation play key roles in many diseases and have been studied in various settings. Comparison and interpretation of measurements at the capillary level, however, is difficult when different techniques are employed and when data on systemic parameters are missing. AIM: To investigate (a) how changes in systemic parameters influence microcirculation and tissue oxygenation; (b) if these changes are detectable at the capillary level, and (c) which systemic parameters must be monitored for reliable interpretation of microcirculatory parameters. METHODS: Assessment of capillary blood flow (CBF) and mucosal oxygen supply (Hb(Sat)O(2), p(muc)O(2)) of the colon in rats by (i) intravital microscopy (IVM), (ii) micro-light guide spectroscopy (EMPHO), and (iii) polarographic micro-catheter probe (LICOX) under (a) physiological conditions, (b) hypovolaemia, and (c) hypoxia. RESULTS: CBF (IVM), Hb(Sat)O(2) (EMPHO) and p(muc)O(2) (LICOX) changed significantly under hypovolaemia, but with different extents. Under hypoxia, CBF did not change, whereas Hb(Sat)O(2) and p(muc)O(2) decreased significantly to 52 and 67% of baseline. Correlation of IVM/EMPHO and IVM/LICOX was poor (r = 0.12; r = 0.03). CONCLUSION: Changes of systemic parameters have significant effects on peripheral tissue oxygenation but may not always be detected at the capillary level. MAP, blood gases and haematocrit must be analysed to correctly interpret microcirculatory parameters. PMID- 17921674 TI - Influence of temporary abdominal wall repair on the intestinal integrity: an experimental study in the rat. AB - BACKGROUND AND AIMS: The aim of this study was to analyze intestinal integrity after temporary abdominal wall repair with absorbable mesh. METHODS: Rats underwent abdominal wall repair with absorbable mesh or sham operation. Myeloperoxidase-positive cells in the intestinal muscularis were histochemically quantified. Intestinal transit was visualized 48 h after surgery. Local and systemic inflammatory response was measured with TNF-alpha and IL-6 ELISA as well as malondialdehyde (MDA) expression in serum and peritoneal fluid. RESULTS: Neutrophil count of the intestinal muscularis revealed that infiltration in the mesh-implanted and in the mesh-free animals 48 h postoperatively was similar. Gastrointestinal transit was similarly unaffected 48 h after surgery, with or without mesh implantation. TNF-alpha, IL-6 and MDA concentration in serum and peritoneal fluid showed no significant differences between the two groups. CONCLUSION: Intestinal contractility and local and systemic inflammatory response remained unaffected. Therefore, absorbable mesh augmentation is a safe and reliable method for temporary repair of the abdominal wall without affecting the intestinal integrity. PMID- 17921675 TI - Impact of male takeover on intra-unit sexual interactions and subsequent interbirth interval in wild Rhinopithecus roxellana. AB - Data were collected on sexual interactions before and after a male takeover of a one-male unit (OMU) of Sichuan snub-nosed monkeys (Rhinopithecus roxellana) in the Qinling Mountains, China. The original unit consisted of an adult male, 2 adult and 2 subadult females, 2 female juveniles and a single infant. Following the takeover, the new resident male copulated with 1 adult female, which was not lactating. Subsequent to the disappearance of her infant, the second (lactating female) entered breeding condition and began to solicit copulation with the new resident male. The subadult females also engaged in matings with the new male. The new resident male was observed mating, on 3 occasions, with females in 2 other OMUs. These are the first observations of sexual behaviour in free-ranging Sichuan snub-nosed monkeys after an OMU takeover. Sexual interactions play an important role in establishing relationships between the new male and resident females in the OMU. PMID- 17921676 TI - Progressive derangement of the T cell compartment in a case of Evans syndrome. AB - BACKGROUND: Evans syndrome (ES) is a rare disorder characterized by combined autoimmune thrombocytopenia and autoimmune hemolytic anemia. Several studies have documented a number of B cell defects, whereas only limited information is currently available about the T cell subset. METHODS: A wide panel of immunological analyses aiming specifically at a quantitative and qualitative evaluation of the T cell compartment was performed in an unusual case of ES. The peripheral distribution of the T cell subsets, the diversity of the T cell receptor (TCR) repertoires, the cytokine profile and the T cell apoptosis have been longitudinally evaluated. RESULTS: On first investigation, flow-cytometric immunophenotyping showed a remarkable alteration of T cell homeostasis with deeply reduced CD4+ naive T cells and recent thymic emigrants. This was seen in association with increased levels of T cell activation and apoptosis. Consistently with these data the cytokine profile was characterized by high interferon-gamma and low interleukin-2 levels. Staining for CD4 and CD25 molecules showed decreased percentages of circulating regulatory T cells according to the autoimmune nature of ES. Finally, restricted TCR repertoires were demonstrated by a skewed TCR beta chain variable (TCRBV) gene usage as well as oligoclonal third complementarity-determining region (CDR3) profiles. A deterioration of the above-mentioned parameters and a worsening of the clinical condition were observed during the follow-up requiring more intensive treatments. CONCLUSION: The demonstration of multiple T cell defects, in addition to providing pathogenetic information, is likely to alter both acute treatment and outcome of ES. PMID- 17921677 TI - Omalizumab treatment associated with Churg-Strauss vasculitis. PMID- 17921678 TI - Spontaneous regression of a herniated cervical disc. PMID- 17921679 TI - Recurrent hemoptysis from diffuse tracheal nodularity due to tracheobronchopathia osteochondroplastica. PMID- 17921680 TI - A 37-year-old married woman with 3-month history of galactorrhoea and irregular menstrual cycle. Diagnosis: multiple endocrine neoplasia type 1 (MEN 1), also known as Wermer's syndrome. PMID- 17921681 TI - Bilateral benign endobronchial schwannomas. PMID- 17921682 TI - Comparison of the erythrocyte sedimentation rate measured by the Micro Test 1 sedimentation analyzer and the conventional Westergren method. AB - BACKGROUND: The erythrocyte sedimentation rate (ESR) remains the most widely used laboratory test for monitoring infections, inflammatory diseases and some types of cancer. Several test methods have been developed recently, and as a result, the safety and reliability of ESR testing procedures have improved. The purpose of this study was the comparison of two methods, the traditional manual Westergren method (reference method of the International Committee on Standardization in Hematology) and a new semiautomated technique, the Micro Test 1 for determining the ESR. SUBJECTS AND METHODS: Blood samples were collected after a night's fasting from 200 hospitalized and ambulatory patients. Undiluted blood samples anticoagulated with K3 EDTA that had Micro Test 1 values ranging from 2-82 mm/h were used for comparison with the Westergren method. RESULTS: Linear regression analysis comparing the Micro Test 1 and the reference method yielded satisfactory correlations and regression for samples (r=0.910; P=0.0001; y=4.91+0.86 x; Sy/x=6.85). A Bland-Altman analysis showed no evidence of systematic bias between the Micro Test 1 and the reference method. CONCLUSION: The Micro Test 1 system was easy to use, had a satisfactory operative practicability, required minimal maintenance, and reduced contact with potential biohazards. PMID- 17921683 TI - Age- and body mass index-dependent relationship between correction of iron deficiency anemia and insulin resistance in non-diabetic premenopausal women. AB - BACKGROUND: No prospective studies have evaluated the effects of correction of iron deficiency anemia on insulin resistance in non-diabetic premenopausal women. We investigated this relationship in 54 non-diabetic premenopausal women with iron deficiency anemia. SUBJECTS AND METHODS: All patients were treated with oral iron preparations. Insulin resistance was calculated with the Homeostasis Model Assessment formula. All patients were dichotomized by the median for age and BMI to assess how the relationship between iron deficiency anemia and insulin resistance was affected by age and BMI. RESULTS: Although the fasting glucose levels did not change meaningfully, statistically significant decreases were found in fasting insulin levels following anemia treatment both in the younger age (<40 years) (P=0.040) women and in the low BMI (<27 kg/m2) (P=0.022) subgroups but not in the older age (>or=40 years) and the high BMI (>or=27 kg/m2) subgroups. Post-treatment fasting insulin levels were positively correlated both with BMI (r=0.386, P=0.004) and post-treatment hemoglobin levels (r=0.285, P=0.036). Regression analysis revealed that the factors affecting post-treatment insulin levels were BMI (P=0.001) and post-treatment hemoglobin levels (P=0.030). CONCLUSION: Our results show that following the correction of iron deficiency anemia, insulin levels and HOMA scores decrease in younger and lean non-diabetic premenopausal women. PMID- 17921684 TI - Impact of hyperglycemia on ischemic stroke mortality in diabetic and non-diabetic patients. AB - BACKGROUND: Previous studies suggest that infarct expansion may be responsible for increased mortality after stroke onset in patients with prolonged stress hyperglycemia. Therefore, we evaluated the influence of prolonged stress hyperglycemia on stroke mortality in patients with and without diabetes. PATIENTS AND METHODS: For 630 stroke patients admitted to the neurological intensive care department within 24 hours of stroke onset, we correlated mean blood glucose levels (MBGL) at admission and 72 hours after admission in diabetic and non diabetic patients with final outcome. Blood glucose levels higher then 6.1 mmol/L (121 mg/dL) were treated as hyperglycemia. RESULTS: Of 630 patients (mean age 71 A+/- 6), 410 were non-diabetic (mortality, 25%) and 220 patients were diabetic (mortality, 20%). All patients who died within 28 days of hospitalization had prolonged hyperglycemia (at admission and after 72 hours, despite insulin therapy). The unadjusted relative risk of in-hospital mortality within 28 days for all stroke patients was 0.68 (95% CI, 0.14-1.9) for non-diabetic patients and 0.39 (95% CI, 0.27- 1.56) for diabetic patients. The unadjusted relative risk of in-hospital mortality within 28 days in ischemic stroke in patients with MBGL > 6.1-8.0 mmol/L (121-144 mg/dL) at admission and after 72 hours was 1.83 (95% CI, 0.41-5.5) for non-diabetic patients and 1.13 (95% CI, 0.78-4.5) for diabetic patients. Non-diabetic patients with hyperglycemia had a 1.7 times higher relative risk of in-hospital 28-day mortality than patients with diabetes. CONCLUSION: Prolonged stress hyperglycemia in ischemic stroke patients increases the risk of in-hospital 28- day mortality, especially in non-diabetic patients. PMID- 17921685 TI - Smoking habits among Saudi female university students: prevalence, influencing factors and risk awareness. PMID- 17921686 TI - The utility of fasting plasma glucose in predicting glycosylated hemoglobin in type 2 diabetes. AB - BACKGROUND: Access to glycosylated hemoglobin (HbA1c) assays in clinical practice remains limited. We investigated the relationship of fasting plasma glucose and HbA1c to determine optimal glucose levels for predicting HbA1c. PATIENTS AND METHODS: We retrospectively analyzed data on 2888 patients with type 2 diabetes mellitus aged >or=20 years using a linear regression of HbA1c against fasting plasma glucose. A receiver-operating characteristic analysis was used to determine optimal cut-points for fasting glucose in relation to HbA1c, area under the curve, sensitivity and specificity, and 95% confidence intervals (CI) for each cut-point. RESULTS: The mean (standard deviation) for the age of patients was 52+/-11.6 years. The average HbA1c was 8.9+/-2.46% and mean fasting plasma glucose was 10.1+/-3.62 mmol/L. The prevalence of HbA1c >or=7.0% and >6.5% was 76% and 82%, respectively. Overall, fasting plasma glucose and HbA1c were linearly correlated (r=0.62, P=0.001). A fasting plasma glucose of >9.0 mmol/L predicted HbA1c >or=7.0% with an area under the curve = 0.807 (95% CI, 0. 0.794 to 0.821), while fasting plasma glucose >8.2 mmol/L predicted HbA1c >6.5%, with an area under the curve = 0.805 (95% CI, 0.791 to 0.818). The sensitivity of both cut-points was 64.5% and 70.7%, the specificity was 82.7% and 76.4%, the positive likelihood ratio was 3.73 and 2.99, and the positive predictive value was 92.2% and 93.2%, respectively. CONCLUSION: When HbA1c determination is not available, fasting plasma glucose levels may be used to identify patients with uncontrolled type 2 diabetes and initiate timely intensification of therapy to avoid long- term complications of diabetes. PMID- 17921687 TI - Painful opthalmoplegia: the Tolosa-Hunt syndrome. PMID- 17921689 TI - Cryptosporidiosis in a renal transplant patient treated with paromomycin. PMID- 17921690 TI - Predicting glucose intolerance with normal fasting plasma glucose by the components of the metabolic syndrome. AB - BACKGROUND: Surprisingly, it is estimated that about half of type 2 diabetics remain undetected. The possible causes may be partly attributable to people with normal fasting plasma glucose (FPG) but abnormal postprandial hyperglycemia. We attempted to develop an effective predictive model by using the metabolic syndrome (MeS) components as parameters to identify such persons. SUBJECTS AND METHODS: All participants received a standard 75-g oral glucose tolerance test, which showed that 106 had normal glucose tolerance, 61 had impaired glucose tolerance, and 6 had diabetes-on-isolated postchallenge hyperglycemia. We tested five models, which included various MeS components. Model 0: FPG; Model 1 (clinical history model): family history (FH), FPG, age and sex; Model 2 (MeS model): Model 1 plus triglycerides, high-density lipoprotein cholesterol, body mass index, systolic blood pressure and diastolic blood pressure; Model 3: Model 2 plus fasting plasma insulin (FPI); Model 4: Model 3 plus homeostasis model assessment of insulin resistance. A receiver-operating characteristic (ROC) curve was used to determine the predictive discrimination of these models. RESULTS: The area under the ROC curve of the Model 0 was significantly larger than the area under the diagonal reference line. All the other 4 models had a larger area under the ROC curve than Model 0. Considering the simplicity and lower cost of Model 2, it would be the best model to use. Nevertheless, Model 3 had the largest area under the ROC curve. CONCLUSION: We demonstrated that Model 2 and 3 have a significantly better predictive discrimination to identify persons with normal FPG at high risk for glucose intolerance. PMID- 17921688 TI - Cryptosporidiosis in Saudi Arabia and neighboring countries. AB - Cryptosporidium is a coccidian protozoan parasite of the intestinal tract that causes severe and sometimes fatal watery diarrhea in immunocompromised patients, and self-limiting but prolonged diarrheal disease in immunocompetent individuals. It exists naturally in animals and can be zoonotic. Although cryptosporidiosis is a significant cause of diarrheal diseases in both developing and developed countries, it is more prevalent in developing countries and in tropical environments. We examined the epidemiology and disease burden of Cryptosporidium in Saudi Arabia and neighboring countries by reviewing 23 published studies of Cryptosporidium and the etiology of diarrhea between 1986 and 2006. The prevalence of Cryptosporidium infection in humans ranged from 1% to 37% with a median of 4%, while in animals it was different for different species of animals and geographic locations of the studies. Most cases of cryptosporidiosis occurred among children less than 7 years of age, and particularly in the first two years of life. The seasonality of Cryptosporidium varied depending on the geographic locations of the studies, but it was generally most prevalent in the rainy season. The most commonly identified species was Cryptosporidium parvum while C. hominis was detected in only one study from Kuwait. The cumulative experience from Saudi Arabia and four neighboring countries (Kuwait, Oman, Jordan and Iraq) suggest that Cryptosporidium is an important cause of diarrhea in humans and cattle. However, the findings of this review also demonstrate the limitations of the available data regarding Cryptosporidium species and strains in circulation in these countries. PMID- 17921691 TI - Experience with 122 consecutive liver transplant procedures at King Faisal Specialist Hospital and Research Center. AB - BACKGROUND: Saudi Arabia is a leading country in the Middle East in the field of deceased-donor liver transplantation (DDLT) and living-donor liver transplantation (LDLT). We present out experience with DDLT and LDLT at King Faisal Specialist Hospital and Research Center (KFSHRC) for the period from April 2001 to January 2007. PATIENTS AND METHODS: We performed 122 LT procedures (77 DDLTs and 45 LDLTs) in 118 patients (4 re-transplants) during this period of time. RESULTS: The number of adult and pediatric procedures was 107 and 11, respectively. The overall male/female ratio was 66/52 and the median age of patients was 43 years (range, 2-63 years). In the DDLT group, the median operating time was 8 hours (range, 4-19), the median blood transfusion was 6 units (range, 0-40), and the median hospital stay was 13 days (range, 6-183). In the DDLT group, after a mean follow-up period of 760 days (range, 2-2085), the overall patient and graft survival rate was 86%. In the LDLT group, the median operating time was 11 hours (range, 7-17), the median blood transfusion was 4 units (range, 0-65), and the median hospital stay was 15 days (range, 7-127). In the LDLT group, and after a mean follow-up period of 685 days (range, 26- 1540), the overall patient and graft survival rates were 90% and 80%, respectively with no significant difference in patient and graft survivals between groups. Biliary complications were significantly higher in LDLT compared to DDLT (P<0.05). Vascular complications were also significantly higher in LDLT compared DDLT (P<0.05). CONCLUSIONS: Both DDLT and LDLT are being successfully performed at KFSHRC with early experience indicating a higher rate of biliary and vascular complications in the LDLT group. PMID- 17921692 TI - Lung cancer in Bahrain (1952-2004). PMID- 17921693 TI - Ibn al-Nafis: discoverer of the pulmonary circulation. PMID- 17921694 TI - Autophagic-lysosomal dysfunction and neurodegeneration in Niemann-Pick Type C mice: lipid starvation or indigestion? AB - Increasing evidence shows that autophagy, particularly macroautophagy, plays a Dr. Jekyll and Mr. Hyde role in determining cell fate; autophagic activity can be protective under certain conditions, whereas it may lead to cell death under others. Niemann-Pick Type C (NPC) disease is an early onset autosomal recessive disorder characterized by accumulation of cholesterol and other lipids in late endosomes/lysosomes. About 95% of the cases are caused by mutations in the NPC1 gene, whereas the remaining 5% are due to mutations in the NPC2 gene. Severe neurodegeneration that accompanies NPC is likely the fatal cause in this disease, although the underlying mechanism remains unclear. Our study shows that autophagic activity is enhanced in Npc1-/- mice, as evidenced by increased levels of LC3-II and the number of autophagic vacuole-like structures. Interestingly, LC3 immunoreactivity co-localizes with filipin-labeled cholesterol clusters inside Purkinje cells. Furthermore, increases in autophagic activity are closely associated with alteration in lysosomal function and protein ubiquitination. In this article, these results are further discussed in the context of autophagic lysosomal function and neuronal survival and degeneration. PMID- 17921695 TI - Genome-wide association scanning highlights two autophagy genes, ATG16L1 and IRGM, as being significantly associated with Crohn's disease. AB - The era of genome-wide association (GWA) scanning has shed new light on the genetic basis of common disease and nowhere is this better illustrated than Crohn's disease (CD). CD is a chronic debilitating inflammatory bowel disease characterized by stricturing and fistula formation. Mainstays of current therapy are immune suppression and surgery. The pathogenesis of CD is poorly understood, but it has long been recognized that both genetic susceptibility and bacterial antigens play important roles. A variety of intracellular bacteria have been postulated to trigger CD, but the evidence for any one organism is equivocal. The current consensus is that commensal gut bacteria provide the drive for CD-related inflammation. Three GWA scans undertaken in the last 6 months have identified 10 new loci demonstrating highly significant and replicated association with CD. Two of the strongest hits implicate genes IRGM and ATG16L1, which encode proteins thought to be critical to the autophagy pathway. The critical next step is functional characterization of the CD-associated genetic variants in IRGM and ATG16L. It seems highly plausible that variation in these genes holds the key to understanding exactly which bacteria drive the intestinal inflammation of CD and the mechanism by which they do this. PMID- 17921696 TI - The non-canonical role of Atg family members as suppressors of innate antiviral immune signaling. AB - Recent research on autophagy clearly demonstrates that the autophagosome-lysosome pathway plays essential roles in elimination of certain pathogens such as group A Streptococcus, Mycobacterium tuberculosis, Listeria monocytogenes, and herpesvirus. (1-4) We have recently found that a key regulator of the autophagic process, the Atg12-Atg5 conjugate, associates with the signaling molecules retinoic acid-inducible gene I (RIG-I) and interferon-beta promoter stimulator 1 (IPS-1), which are essential for recognition of RNA virus infection and which transmit signals to upregulate type I interferons (IFNs). Interestingly, the Atg12-Atg5 conjugate seemed to negatively regulate the type I IFN modulating pathway through direct interaction with caspase recruitment domains (CARDs) presented by RIG-1 and IPS-1.(5) Thus, in contrast to the bactericidal properties of autophagic processes, the autophagy regulator (the Atg12-Atg5 conjugate) appeared to promote RNA virus replication by inhibiting innate anti-virus immune responses. In this addendum, we discuss the non-canonical role of the Atg12-Atg5 conjugate as a suppressor of innate immune responses. PMID- 17921697 TI - Pex14 is the sole component of the peroxisomal translocon that is required for pexophagy. AB - Pex14 was initially identified as a peroxisomal membrane protein that is involved in docking of the soluble receptor proteins Pex5 and Pex7, which are required for import of PTS1- or PTS2-containing peroxisomal matrix proteins. However, Hansenula polymorpha Pex14 is also required for selective degradation of peroxisomes (pexophagy). Previously we showed that Pex1, Pex4, Pex6 and Pex8 are not required for this process. Here we show that also in the absence of various other peroxins, namely Pex2, Pex10, Pex12, Pex13 and Pex17, pexophagy can normally occur. These peroxins are, like Pex14, components of the peroxisomal translocon. Our data confirm that Pex14 is the sole peroxin that has a unique dual function in two apparent opposite processes, namely peroxisome formation and selective degradation. PMID- 17921698 TI - Activation of polyamine catabolism by N1, N11-diethylnorspermine alters the cellular localization of mTOR and downregulates mTOR protein level in glioblastoma cells. AB - N(1), N(11)-Diethylnorspermine (DENSPM) is a spermine analog and prototype anti cancer drug that depletes cellular polyamine, increases cellular oxidative stress through the generation of H(2)O(2) and induces the death of multiple types of cancer cells. However, the survival pathways perturbed by DENSPM are uncertain. To identify these pathways, we examined a series of proteins in the phosphoinositide 3-kinase /AKT/mammalian target of rapamycin (PI3K/AKT/mTOR) pathways in glioblastoma cell lines before and after treatment with DENSPM. We found that DENSPM did not change the protein levels of PI3K but did reduce the levels of AKT, phosphorylated AKT, mTOR, phosphorylated mTOR, p70(S6K), phosphorylated p70(S6K), 4E-BP1, phosphorylated 4E-BP1 and eIF-4B proteins. From this it appears that DENSPM directly targets the mTOR protein level in these glioblastoma cells by inhibiting mTOR-mediated protein synthesis. Immunofluorescence analysis of mTOR showed that DENSPM sequestered mTOR in the perinuclear region of the cells. We also detected a marked collapse of microtubules in U87 cells and a detachment of cells in a process resembling anoikis. We further showed that the levels of many proteins regulating cell growth and cell adhesion were downregulated, suggesting a broad effect of DENSPM on mTOR-mediated protein synthesis. We conclude that the activation of polyamine catabolism alters the cellular location of mTOR, thus negatively affecting mTOR mediated protein synthesis and leading to the death of glioblastoma cells. PMID- 17921699 TI - Novel splice isoforms of STRADalpha differentially affect LKB1 activity, complex assembly and subcellular localization. AB - STRADalpha is a pseudokinase that forms a heterotrimeric complex with the scaffolding protein MO25 and the tumor suppressor serine threonine protein kinase LKB1. Mutations in the LKB1 gene are responsible for the Peutz-Jeghers Syndrome (PJS) characterized by a predisposition to hamartomatous polyps and hyperpigmentation of the buccal mucosa. Mutations in LKB1 have also been observed in some sporadic tumours unrelated to PJS. The LKB1/STRAD/MO25 complex is involved in the regulation of numerous signaling pathways including metabolism, proliferation and cellular polarity of human intestinal epithelial cells. Cell polarization, together with tissue-restricted transcription, represents the main feature of enterocyte differentiation. Since a full-length STRADalpha transcript has not been identified thus far in these cells, we evaluated the expression of endogenous STRADalpha in five colorectal cancer cell lines characterized by their diverse ability to differentiate in vitro. We report herein the discovery of several novel splice isoforms of STRADalpha that differentially affect the kinase activity, complex assembly, subcellular localization of LKB1 and the activation of the LKB1-dependent AMPK pathway. PMID- 17921701 TI - Following the tracks of AKT1 gene. AB - The AKT or PKB family of protein kinases is one of the best characterized targets of phosphoinositide 3-kinases or PI3Ks. The AKT/PKB signal transduction pathway regulates many growth and survival mechanisms including transcription, cell cycle progression, metabolism, apoptosis, invasion and metastasis. Recently, Carpten et al. (Nature 2007;448:439-44) reported a very interesting study on the isoform AKT1 (PKBalpha). They described a novel point mutation (E17K) in the pleckstrin homology domain (PHD) of the AKT1 gene in human breast, colorectal and ovarian cancers, and demonstrated that it induces leukemia in mice. Here we give a critical appraisal of this finding and underline its clinical impact. We also discuss possible interventions for therapeutic development of AKT inhibitors to treat human cancer. PMID- 17921700 TI - Pre-clinical evaluation of 1-nitroacridine derived chemotherapeutic agent that has preferential cytotoxic activity towards prostate cancer. AB - Chemotherapy in prostate cancer (CaP) even as an adjunct has not been a success. In this communication, we report the pre-clinical efficacy of a nitroacridine derivative, C-1748 (9[2'-hydroxyethylamino]-4-methyl-1-nitroacridine) in CaP cell culture and human xenograft animal models. C-1748, a DNA intercalating agent has been derived from its precursor C-857 that was a potent anti-cancer drug, but failed clinical development due to "high" systemic toxicities. Chemical modifications such as the introduction of a "methyl" group imparted novel properties, the most interesting of which is the difference in the IC(50) values between LnCaP (22.5 nM), a CaP cell line and HL-60, a leukemia cell line (>100 nM). Using gammaH2AX as an intervention marker of DNA double strand breaks, we concluded that C-1748 is more efficacious in CaP cells than in HL-60 cells. In hormone dependent cells, the androgen receptor (AR) was identified as an additional target of C-1748. In xenograft studies, administration of C-1748 intra peritoneally inhibited tumor growth by 80-90% with minimal toxicity. These studies identify C-1748 as a novel acridine drug that has a high therapeutic index and low cytotoxicity on myelocytic cells with potential for clinical development. PMID- 17921702 TI - An octavalent lyme disease vaccine induces antibodies that recognize all incorporated OspC type-specific sequences. AB - Lyme disease is the most common vector-borne disease in North America and Europe and, if untreated, has significant arthritic, cardiac, dermatological and neurological sequelae. There is no currently available human Lyme disease vaccine. Outer surface protein C, because of its antigenicity, protective ability, and expression characteristics has emerged as a promising second generation vaccine candidate; however, significant sequence heterogeneity has impeded its development. Analyses of OspC sequences have revealed the existence of stable phylogenetic clusters or types, and that the type-defining sequence variation occurs within defined domains of the protein. Recent data indicating that immunodominant, and potentially protective OspC epitopes are located in these hypervariable regions has allowed development of a tetravalent, epitope based, chimeric vaccine. In this report, we have extended that previously described tetravalent construct to include four additional OspC types. We demonstrate that the construct is highly immunogenic, and elicits type-specific antibodies that recognize each of the eight incorporated OspC type-specific epitopes. Antibody raised to the octavalent construct readily binds to the surface of strains expressing each component OspC type, indicating that the incorporated epitopes are presented on the surface of intact cells. In addition, the construct elicits antibody isotypes associated with complement-dependent bactericidal activity. These results represent an important step forward in the design of a broadly protective polyvalent OspC-based Lyme disease vaccine. PMID- 17921703 TI - Characterization of the key antigenic components and pre-clinical immune responses to a meningococcal disease vaccine based on Neisseria lactamica outer membrane vesicles. AB - Serogroup B strains are now responsible for over 80% of meningococcal disease in the UK and no suitable vaccine is available that confers universal protection against all serogroup B strains. Neisseria lactamica shares many antigens with the meningococcus, except capsule and the surface protein PorA. Many of these antigens are thought to be responsible for providing cross-protective immunity to meningococcal disease. We have developed an N. lactamica vaccine using methods developed for meningococcal outer membrane vesicle (OMV) vaccines. The major antigenic components were identified by excision of 11 major protein bands from an SDS-PAGE gel, followed by mass spectrometric identification. These bands contained at least 22 proteins identified from an unassembled N. lactamica genome, 15 of which having orthologues in published pathogenic Neisseria genomes. Western blotting revealed that most of these bands were immunogenic, and antibodies to these proteins generally cross-reacted with N. meningitidis proteins. Sera from mice and rabbits immunized with either N. lactamica or N. meningitidis OMVs produced comparable cross-reactive ELISA titres against OMVs prepared from a panel of diverse meningococcal strains. Mice immunized with either N. meningitidis or N. lactamica OMVs showed no detectable serum bactericidal activity against the panel of target strains except N. meningitidis OMV sera against the homologous strain. Similarly, rabbit antisera to N. lactamica OMVs elicited little or no bactericidal antibodies against the panel of serogroup B meningococcal strains. However, such antisera did mediate opsonophagocytosis, suggestingthat this may did mediate opsonophagocytosis, suggesting that this may be a mechanism by which this vaccine protects in a mouse model of meningococcal bacteraemia. PMID- 17921704 TI - National surgical patterns of care: regional lymphadenectomy of breast sarcomas. AB - PURPOSE: Soft tissue sarcomas (STS) of the breast are rare tumors with few large experiences available regarding their optimal management. Although assessment of the draining lymphatics is not a component of standard surgical treatment of soft tissue sarcomas, we hypothesized that the management of breast STS may present a unique situation: given their experience with the more common epithelial lesions of the breast, surgeons may be more prone to sample lymph nodes (LNs) in these patients than would otherwise be indicated based on histologic subtype. We reviewed national surgical patterns of care for regional lymphadenectomy for breast STS using the Surveillance Epidemiology & End Results (SEER) registry. MATERIALS/METHODS: SEER data for LN evaluation are available from 1988. The public-access SEER registry was queried for patients presenting between 1988 and 2002 with breast STS. Data were collated by number of LNs examined and further analyzed by histology, grade, and size of the primary lesion where available. For 5-year relative survival calculations, 210 patients with STS from 1988 to 1998 were analyzed. Relative survival in SEER is defined as the observed survival of a cohort divided by the observed survival of an age-, race-, and gender-matched cohort without disease; it thus serves as a surrogate for disease-free survival. RESULTS: A total of 333 cases of STS were identified for the 15-year period; 39% of patients underwent some degree of regional lymphadenectomy, and the median number of LNs examined in these patients was 10 (range, 1-34). Of all patients, 17.4% underwent lymphadenectomy of 10 LNs or more. More axillary procedures were performed as grade increased (30.8% grade I/II, 53.8% grade III/IV ([chi]2 = 10.8; P < 0.001). The percentage of patients undergoing radical or modified radical mastectomy did not change significantly over the 15-year period (chi2 = 1.25; P < 1.0), although procedures sampling the axilla were significantly more frequent when lesions were larger than 5 cm (chi2 = 4.14; P < 0.05). None of 6 patients with positive LNs survived 5 years. Examination of nodes did not impact survival: 117 patients without LNs sampled had 5-year related survival of 68.7% (SE = 5.5%); 89 patients with LN sampling had 64.3% 5-year related survival (SE = 5.8%). CONCLUSION: Lymphadenectomy is frequently performed despite nodal metastasis being an infrequent occurrence in breast STS. LN metastases were rarely identified and lymphadenectomy did not confer a survival benefit. PMID- 17921705 TI - Local recurrence after mastectomy in patients with T3pN0 breast carcinoma treated without postoperative radiation therapy. AB - OBJECTIVE: The need for comprehensive adjuvant radiotherapy in patients with T3pN0 breast cancer is controversial. This retrospective analysis was performed to assess the frequency of local and distant recurrence in patients treated with mastectomy without postoperative radiation. MATERIALS AND METHODS: A single institution database of 2362 patients with breast carcinoma treated from 1974 to 1994 yielded 101 patients who had T3pN0 disease and did not receive chest wall or nodal irradiation. The median follow-up time was 93 months (range, 10-256 months). Sites of first failure were categorized as isolated chest wall (CWF), regional lymph nodes (RNF, which in this case were considered to be either axillary or supraclavicular), or distant sites (DF). CWF and/or RNF were considered local recurrences. Patients with simultaneous CWF and DF or RNF and DF were scored as DF. A comparison was made to 286 T2pN0 patients, also treated between 1974 and 1994. RESULTS: Twenty-two T3N0 patients developed recurrent disease. Site of first recurrence was isolated local recurrence in 11 patients and distant in 11 patients. Four patients had simultaneous local and distant recurrences. Site of isolated local recurrence was CWF in 5 patients and RNF in 6 patients. Median tumor size was 6 cm (range, 5-10.5 cm). There was no difference in local recurrence for tumor sizes < or =7 cm versus >7 cm (P = 0.07). The crude recurrence rate for T3pN0 patients treated by mastectomy was similar to T2pN0 patients treated in similar fashion (P = 0.3). CONCLUSION: The risk of isolated local recurrence in patients with T3pN0 breast cancer and negative margins is moderately low and similar to T2pN0 patients. These results suggest that routine use of postoperative chest wall and nodal irradiation in all T3pN0 patients may not be required. PMID- 17921706 TI - Predicting the risk of local recurrence in patients with breast cancer: an approach to a new computer-based predictive tool. AB - OBJECTIVE: To develop a new web-based tool, designated IBTR!, which integrates prognostic factors for local recurrence (LR) into a model to predict the 10-year risk of LR after breast conserving surgery (BCS) with or without radiation therapy (RT) with the goal of assisting with patient counseling and medical decision-making. METHODS: All available randomized trials of BCS alone versus BCS plus RT, meta-analyses, and institutional reports were reviewed to identify the principal prognostic factors for LR after breast-conserving therapy. Patient age, margin status, lymphovascular invasion (LVI), tumor size, tumor grade, use of chemotherapy, and use of hormonal therapy were found to consistently and significantly impact LR across multiple studies. Based upon a composite analysis of the relevant published randomized and nonrandomized studies, relative risk (RR) ratios were estimated and assigned to each prognostic category. These RR ratios were entered into a mathematical model with the 10-year baseline rates of recurrence with and without RT, 7% and 24%, respectively, to predict patient specific LR risk. RESULTS: Individual data entered into this computer model with regards to patient age, margin status, LVI, tumor size, tumor grade, use of chemotherapy, and use of hormonal therapy will generate patient-specific predicted 10-year LR risk with and without RT. A graphic representation of the relative risk reduction with RT will also be displayed alongside the numerical display. CONCLUSION: IBTR! is a first attempt at a computer model incorporating LR prognostic factors in an evidence-based fashion to predict individual LR risk and the potential additional benefit from RT. PMID- 17921707 TI - Dose escalating study of biweekly gemcitabine and carboplatin in patients with advanced cancer. AB - Combination chemotherapy with gemcitabine and carboplatin administered on a 3 week cycle is used commonly in the treatment of cancer. The purpose of our study was to establish a safe dose of combined gemcitabine and carboplatin when administered on a biweekly schedule to patients with advanced solid tumors. Gemcitabine was given intravenously over 30 minutes followed by carboplatin also given intravenously over 30 minutes once every 2 weeks (one cycle). Five dose levels were examined, ranging from gemcitabine at 1250 mg/m2 to 2000 mg/m2 and carboplatin at an area under the curve of 2.5 to 3.0. Twenty-six patients were studied (18 male and 8 female) with a median age of 57 years (range, 41-83 years); ECOG performance status was 0 or 1 in 22 patients (85%); median number of prior chemotherapy regimens was 2 (range, 0-4); median number of cycles administered per patient was 3 (range, 1-9) with a total of 89 cycles. Two dose limiting toxicities were observed. Delay in treatment was seen in a total of 8 cycles with 6 of the delays due to myelosuppression. Grade 3 or 4 hematologic toxicity rates were as follows: anemia in one cycle (1%), neutropenia in 13 cycles (15%), and thrombocytopenia in one cycle of chemotherapy (1%). There were no hospitalizations for neutropenic fever. Mild fatigue was the most common nonhematologic toxicity. The median time to progression was 40 days (mean, 49 days; range, 4-133 days). Of the 21 evaluable patients, partial response or stable disease was observed in 11 (42%). The maximum tested dose of gemcitabine at 2000 mg/m2 and carboplatin at area under the curve of 3.0 was well tolerated on a biweekly schedule. Our findings indicate that further investigation of this biweekly regimen is warranted in patients with advanced cancer. PMID- 17921708 TI - Phase I study of biweekly paclitaxel and carboplatin for frail patients with advanced non-small cell lung cancer. AB - OBJECTIVE: The aim of this phase I study was to determine the maximum tolerated dose (MTD) and to evaluate the toxicity of the biweekly paclitaxel and carboplatin combination chemotherapy in frail patients with advanced non-small cell lung cancer (NSCLC). METHOD: Seventeen unresectable and previously untreated NSCLC patients participated in this study. Frail patients (>80 years old with PS 0-1 and adequate organ functions, or <80 years old with PS 2 or <80 with PS 0-1 and one inadequate organ function of bone marrow, liver, or kidney) of NSCLC were enrolled in this study. Both paclitaxel and carboplatin were administered on day 1 and repeated biweekly. The starting dose of paclitaxel 120 mg/m2 and carboplatin was AUC 3 /d on day 1 every 2 weeks for 4 to 8 cycles. RESULTS: Seventeen frail patients received a total of 1 to 10 cycles. The major hematologic toxicity was neutropenia. Nonhematologic toxicities, including numbness and constipation, were generally mild and reversible. The MTD of paclitaxel were determined as 140 mg/m2. The recommended phase II study dose of paclitaxel was determined as 120 mg/m2 with area under the curve 3 biweekly for 1 day. An objective response was observed in 8 of 17 patients (47%) with frail patients. CONCLUSION: The combination chemotherapy of biweekly paclitaxel and carboplatin appears to be tolerable and sufficiently effective in frail patients with NSCLC. A phase II trial is underway. PMID- 17921709 TI - Prognostic significance of blood transfusions in patients with esophageal cancer treated with combined chemoradiotherapy. AB - OBJECTIVE: Anemia occurs commonly in patients with esophageal cancer. This study evaluates the effect of blood transfusion on survival outcomes in patients with esophageal cancer treated with combined chemoradiotherapy (CRT). PATIENTS AND METHODS: Fifty-six consecutive patients with unresectable esophageal cancer received 50 Gy in 25 fractions over 5 weeks concurrent with cycles 2 and 3 of cisplatin and 5-fluorouracil chemotherapy. Data on hemoglobin before and during radiation therapy (RT) and blood transfusion use were abstracted by chart review. Each patient had a blood count before every chemotherapy cycle, and the test was repeated if the blood count was low. Five-year Kaplan-Meier overall survival (OS) and relapse-free survival (RFS) estimates were compared according to pre-RT hemoglobin levels and transfusion use. Multivariate analysis using Cox regression modeling was performed to determine the prognostic significance of pre-RT hemoglobin and transfusion use on survival outcomes. RESULTS: The 5-year OS and RFS rates were 30% and 37%, respectively. Seventeen patients (30%) received transfusions during CRT. Among 18 patients (32%) with a hemoglobin of < or =12 g/dL at the start of RT, 9 received transfusions. Pre-RT hemoglobin levels of < or =12 g/dL were strongly associated with the use of blood transfusions (P = 0.03). Five-year Kaplan-Meier OS was 65% versus 21% in patients treated with, versus without, a transfusion (P = 0.006). On multivariate analysis, the use of blood transfusion was associated with improved OS (hazard ratio, 0.26; 95% confidence interval, 0.09-0.75, P = 0.01). CONCLUSIONS: The use of blood transfusion is a significant treatment-related factor associated with improved survival in patients undergoing CRT for esophageal cancer. PMID- 17921710 TI - A phase I dose escalation study of biweekly gemcitabine and carboplatin in completely resected stage IB-IIIA nonsmall cell lung cancer. AB - OBJECTIVE: We conducted a phase I dose escalation study to determine the maximum tolerated dose, recommended dose, and safety profile of a biweekly gemcitabine and carboplatin combination regimen in the treatment of patients with completely resected nonsmall cell lung cancer (NSCLC). PATIENTS AND METHODS: Patients with completely resected pathologically documented stage IB, II, or IIIA NSCLC, performance status (ECOG) 0-1, with adequate bone marrow, renal, liver, and cardiac functions, were treated with gemcitabine and carboplatin. The starting dose was gemcitabine 800 mg/m2 on days 1 and 15 and carboplatin area under the time-concentration curve (AUC) 4 mg/mL/min on day 1. Gemcitabine was increased to 1000 mg/m2 (level 3). Carboplatin was increased to AUC 5 (level 2, 3). The regimen was performed every 4 weeks. The dose-limiting toxicity of the regimen was assessed during the first chemotherapy cycle. RESULTS: Nine patients were enrolled in this study. All patients were assessed for safety. Grade 3 leukopenia occurred in 1 patient (11%) and grade 3/4 neutropenia occurred in 3 patients (33%). No other grade 3/4 toxicity was observed. No dose-limiting toxicity was experienced at dose levels 1, 2, and 3 of this schedule. CONCLUSION: Maximum tolerated dose was not reached in this study. Considering treatment continuation, the recommended dose for a phase II study is gemcitabine 1000 mg/m2 on days 1 and 15 and carboplatin AUC 5 on day 1, every 4 weeks. Biweekly administration of gemcitabine and carboplatin is a feasible and well-tolerated regimen for the treatment of patients with completely resected NSCLC as adjuvant chemotherapy. PMID- 17921711 TI - The use of radiolabeled somatostatin analog scintigraphy in the staging of small cell lung cancer patients. AB - OBJECTIVES: The prognosis of patients with small cell lung cancer (SCLC) is dismal with a median survival not exceeding 18 months. For the fact that the tumor stage remains the most significant prognostic factor, efforts have been made to improve its accuracy. We evaluated the role of somatostatin receptor scintigraphy (SRS) in the diagnosis and initial staging of SCLC in comparison with the conventional staging procedure. METHODS: We administered radiolabeled somatostatin analog Indium 111 (111In)-diethylenetriamine pentaacetic acid (also known as Indium In-111 pentetreotide or OctreoScan) in 32 newly diagnosed patients with SCLC, 19 of which of had limited disease and 13 of which had extensive disease. All patients had been previously examined with other imaging modalities, specifically CT and/or MRI. RESULTS: Staging with 111In-OctreoScan successfully located the primary tumor site with a sensitivity of 92%. Although detection of mediastinal lymph node dissemination was also relatively high (83%), the SRS failed to detect most of the metastatic lesions outside the thorax (9 of 36, 25%), while its sensitivity for the detection of malignant lesions in the liver, adrenals, and bones, was 56%, 33% and 17%, respectively. CONCLUSIONS: Although 111In-OctreoScan may be used in addition to current SCLC staging methods, there are insufficient data for maintaining that SRS may replace conventional staging. PMID- 17921712 TI - Paclitaxel, carboplatin, 5-fluorouracil, and radiation for locally advanced esophageal cancer: phase II results of preliminary pharmacologic and molecular efforts to mitigate toxicity and predict outcomes: North Central Cancer Treatment Group (N0044). AB - PURPOSE: Locally advanced esophageal cancer is challenging to treat. This study tested a 3-drug, multimodality approach. Chemotherapy dose reductions, the addition of amifostine, and pharmacogenetics were tested in an effort to mitigate toxicity and predict outcomes. PATIENTS AND METHODS: This phase II trial tested chemotherapy (carboplatin area under the curve = 4 [lower dose than that from Meluch et al] on day 1 and 22, 5-fluorouracil 225 mg/m2 per day continuous infusion on days 1 to 42, and paclitaxel 200 mg/m2 on days 1 and 22) with concomitant radiation 4500 cG for locally advanced esophageal cancer. Esophagectomy was scheduled after radiation. Amifostine 500 mg subcutaneously before radiation was given to the first 19 patients. RESULTS: Among 54 eligible patients, the pathologic complete response rate was 35% (95% confidence interval [CI], 23%-49%). Median survival was 21.2 months (95% CI, 13.6-37.6 months), and median time to cancer recurrence/progression 19 months (95% CI, 11.4-44.6 months). Nearly all patients (94%) suffered at least one grade 3 or worse adverse event, including 3 treatment-related deaths. Amifostine was discontinued because of one of these deaths. There was no statistically significant difference in the rates of severe adverse events among patients who received amifostine and those who did not. Genotyping for polymorphisms of dihydropyrimidine dehydrogenase, cytochrome P3A4, and glutathione-S-transferase did not predict tumor response or severe adverse events. CONCLUSIONS: This 3-drug, multimodality approach yielded a pathologic complete response rate of 35%, but the severe adverse event rate was high. Utilizing amifostine to reduce toxicity or employing molecular approaches to predict outcomes did not show promise. PMID- 17921713 TI - Treatment outcomes of radiotherapy for patients with stage I esophageal cancer: a single institute experience. AB - OBJECTIVES: To evaluate the effectiveness of radiotherapy for patients with stage I esophageal cancer. METHODS: From 1995 to 2005, 34 patients with superficial thoracic esophageal squamous cell carcinoma without lymph node metastasis were treated with radiotherapy at our institute. Patient characteristics were as follows: median age, 68 years; range, 47 to 84 years; male:female, 33:1; performance status, 0/1/2/3 = 23/9/1/1; T1a/T1b = 4/30. Median fraction and total doses of external irradiation given were 2.0 Gy and 60 Gy, respectively. Twenty three patients had local irradiation to the primary lesion only; the remaining 11 patients received regional field irradiation, including one or 2 regional lymph node areas. Only one patient received adjuvant intracavitary radiotherapy after radiotherapy. Thirty of the patients received concurrent chemotherapy, most of the regimens of which included cisplatin and 5-fluorouracil. Adjuvant chemotherapy was performed in 5 patients. Fifteen patients (44%) had coexisting malignancies. The median follow-up was 38 months (range, 8-107 months). RESULTS: The 5-year overall, cause-specific, and local-progression-free survival rates were 50.4%, 91.2%, and 81.1%, respectively. Of the 34 patients, 6 had in-field local recurrence, and one had recurrence in the esophagus outside of the irradiated field. Two patients experienced regional lymph node metastasis outside of the irradiated field, and one had distant metastasis. Two patients received salvage surgery, and 4 patients received salvage intracavitary radiotherapy after local recurrence. A late toxicity greater than grade 2 was not observed in any patient. CONCLUSIONS: Radiotherapy is an effective treatment modality for patients with stage I esophageal cancer. PMID- 17921714 TI - Prostate high dose rate brachytherapy in a free-standing cancer center setting. AB - OBJECTIVES: To determine the safety and acute tolerance of performing prostate high dose rate (HDR) brachytherapy in a free-standing cancer center setting, and to report our approach in delivering such treatment. METHODS: Patients treated with prostate HDR from the inception of our program through December 2004 were included. Disease- and treatment-related information was gathered from chart review. Acute side effects were obtained from both chart review and questioning the patient at follow-up visits. RESULTS: A total of 234 patients were treated with 459 HDR needle insertions. Nearly all patients were treated with external beam irradiation, and the majority also received adjuvant hormonal manipulation. There were no adverse events related to transport to the free-standing cancer center. Treatment was well tolerated, with only 3.7% of procedures developing retention requiring catheter placement; 20% of patients reported significant postimplant dysuria, mean duration of any dysuria being 7.5 days. Postimplant retention and length of any dysuria were not found to be related to any of the analyzed factors. Severity of dysuria was found to be related only to timing of external beam. Postimplant retention was more frequent for some physicians, but this finding was not statistically significant. CONCLUSIONS: Prostate HDR brachytherapy can be delivered safely and effectively in an ambulatory free standing cancer center setting when done by a urology and radiation oncology team working in close conjunction. Treatment combining HDR brachytherapy as described with external beam radiation, with or without androgen manipulation, is well tolerated by the patient. PMID- 17921715 TI - COX-2 and NF-KB overexpression is common in pancreatic cancer but does not predict for COX-2 inhibitors activity in combination with gemcitabine and oxaliplatin. AB - OBJECTIVE: The attempt to improve therapeutic results in pancreatic carcinoma has recently focused on the emerging role of molecular biology. We investigated the role of COX-2 and NF-KB expression in relation to the use of a COX-2 inhibitor (celecoxib) associated to gemcitabine and oxaliplatin in pancreatic cancer. METHODS: Forty-four patients with histologically or cytologically verified, locally advanced unresectable and/or metastatic pancreatic carcinoma were eligible for the study. RESULTS: Thirty-three patients (75%) assumed celecoxib for all their treatment period. Treatment was repeated every 2 weeks, until there was evidence of disease progression, patient refusal, or unacceptable toxicity. Efficacy was assessed according to tumor response, clinical benefit, and time related parameters. Five patients had a partial response, 24 had a stable disease, and 15 had a disease progression, for an overall response rate of 11%. Biochemical response rate based on CA 19.9 levels showed 2 complete and 10 partial responses, whereas 31 patients presented no changes of CA 19.9 levels. COX-2 protein expression was found in 30 tumors, while a moderate or weak/absent expression was present in 10 patients. Sixteen tumors showed a strong expression for NF-KB, 4 a moderate expression, and 5 a weak/absent expression. CONCLUSION: The use of a COX-2 inhibitor does not add any valuable activity to a gemcitabine/oxaliplatin combination, even in patients with COX-2 and NF-KB overexpressing tumors. PMID- 17921716 TI - Second primary tumors following tobacco dependence treatments among head and neck cancer patients. AB - OBJECTIVES: To estimate the cumulative percentage of second primary tumors (SPTs) in head and neck (H&N) cancer patients and primary cancers in general Nicotine Dependence Center (NDC) population controls following tobacco dependence consultation seen between 1988 and 2001. METHODS: A 1:1 matched pair design and a stratified Cox proportional hazard model were used. General NDC population controls were matched on age, gender, and NDC consult (type and date) to the H&N cancer patients. The study population included 101 H&N cancer patients (66 male, 35 female) with mean (SD) age of 58.7 (10.1) years. RESULTS: Baseline demographics and length of follow-up were similar between groups. However, H&N cancer patients smoked more cigarettes per day than controls (P < 0.003). For H&N cancer patients, the median time from initial H&N cancer diagnosis to NDC consult was 7 months (range, 0-292 months). Following the NDC consult, 27 H&N cancer patients developed 32 SPTs; whereas among the controls, 12 patients developed 12 other cancers (P = 0.013). There was no difference in the development of non tobacco-related cancers (P = 0.273). However, H&N cancer patients were more likely to develop tobacco-related cancers (P = 0.01). Furthermore, there was a trend where the H&N cancer patients who continued to use tobacco were more likely to develop tobacco-related cancers than those who remained abstinent (P = 0.10). CONCLUSIONS: These findings confirm that H&N cancer patients are more prone to the development of tobacco-related cancers. Further, these findings suggest that H&N cancer patients who stop using tobacco are able to decrease the development of tobacco-related SPTs. PMID- 17921718 TI - The influence of mentorship on research productivity in oncology. AB - BACKGROUND: This study evaluates the impact of mentors in research productivity in oncology. METHODS: Two electronic surveys were sent out to 1009 oncologists who attended educational workshops between 1996 and 2004. RESULTS: Response rate was 41.4% (339 of 818). Respondents with mentors are more currently engaged in academic research than those without mentors. Mentorship status did not influence on self-reported publication record or on becoming principal investigators, even when adjusted for other factors. CONCLUSIONS: Mentorship is valuable to oncologists in enhancing their research experiences. In this selected group, mentorship has effects on current involvement in academic research but not on self-reported publication. PMID- 17921717 TI - Chemotherapy and survival for patients with multiple myeloma: findings from a large nationwide and population-based cohort. AB - OBJECTIVE: To assess the patterns of chemotherapy use for patients with multiple myeloma and to determine if chemotherapy is effective in prolonging survival outside the clinical trial settings. METHODS: We studied a nationwide and population-based retrospective cohort of 4902 patients > or =65 years of age with stage II or III multiple myeloma from 1992 to 1999, identified from the Surveillance, Epidemiology, and End-Results-Medicare data. Multivariate logistic regression was used to estimate the odds ratio of receiving chemotherapy and Cox proportional hazard model was used to estimate the hazard ratio of mortality associated with chemotherapy. RESULTS: Of 4902 patients with stage II or III multiple myeloma, 52.0% received chemotherapy during the course of the disease. The receipt of chemotherapy decreased significantly with age from 65.7% in the 65 to 69-year age group to 34.3% in those > or =80 years. Blacks (47.6%) were less likely to receive chemotherapy than whites (52.8%). Use of chemotherapy decreased significantly with comorbidity scores and increased over time. Risk of all-cause mortality was significantly reduced in patients who received chemotherapy compared with those who did not (adjusted hazard ratio = 0.65; 95% confidence interval = 0.61-0.69). A similar pattern as observed for myeloma-specific mortality (0.61; 0.56-0.67). Survival benefit increased with increasing cycles of chemotherapy (P < 0.001 for trend) and was significant across different age groups, gender, ethnic groups, and comorbidity scores. CONCLUSION: Chemotherapy was significantly associated with increased survival in patients with multiple myeloma outside the clinical trial settings. This survival benefit was significant across different groups by age, gender, race, and comorbidity. A substantial number of patients with multiple myeloma did not receive chemotherapy. PMID- 17921719 TI - The many faces of surgical margins. AB - Most variables associated with survival after cancer surgery are fixed when we see the patient. One variable over which we have control is the surgical margin. We begin by reviewing the definitions of a positive surgical margin, and then explore potential inaccuracies in obtaining a biopsy and reasons for recurrence after obtaining a free surgical margin. Research in improving the diagnostic accuracy of surgical margins is discussed. Finally, the prognostic significance of surgical margins is reviewed. PMID- 17921720 TI - Inverted papilloma of the nasal cavity and paranasal sinuses. AB - PURPOSE: To discuss the treatment and outcomes of inverted papilloma of the nasal cavity and paranasal sinuses. METHODS: Review of the pertinent literature. RESULTS: Inverted papilloma is a benign, locally aggressive neoplasm that arises in the nasal cavity and is associated with squamous cell carcinoma in approximately 5% of patients. Squamous cell carcinoma may be present with inverted papilloma at the initial diagnosis or it may occur metachronously after prior treatment. Surgery is the primary treatment of inverted papilloma. The likelihood of local recurrence varies from less than 5% to over 50%, depending on the extent of resection. There is likely no significant difference in the risk of local recurrence after open compared with endoscopic resection. The probability of local recurrence and/or death from tumor is increased if inverted papilloma is associated with squamous cell carcinoma. The likelihood of cure is approximately 50% when malignancy is present and postoperative radiotherapy should be considered for the majority of patients. A small subset of patients with inverted papilloma present with incompletely resectable disease. Definitive radiotherapy using doses between 65 and 70 Gy will locally control gross disease in the majority of patients. CONCLUSION: The preferred treatment of inverted papilloma is surgery; postoperative radiotherapy is added if it is associated with squamous cell carcinoma. The likelihood of local recurrence after surgery for inverted papilloma may be substantial and varies with the extent of resection. Definitive radiotherapy may be used to successfully treat patients with incompletely resectable inverted papilloma. PMID- 17921721 TI - Idiopathic granulomatous lobular mastitis masquerading as a breast abscess and breast carcinoma. PMID- 17921722 TI - Metastatic adenocarcinoma in the cerebellopontine angle mimicking facial nerve Schwannoma. PMID- 17921723 TI - Cancer stem cells: implications for cancer treatment and prevention. PMID- 17921724 TI - An update on treatment advances for the first-line therapy of metastatic colorectal cancer. AB - The treatment of metastatic colorectal cancer (mCRC) has evolved significantly over the past 10 years. For nearly 40 years, the fluoropyrimidine 5-fluorouracil (5-FU) was the only agent to be used for advanced metastatic disease. However, since the mid-1990s, the chemotherapy treatment options for patients with mCRC have been greatly facilitated with the introduction of several new cytotoxic and biologic agents. In particular, combination regimens that incorporate infusional schedules of 5-FU in combination with oxaliplatin (FOLFOX) and/or irinotecan (FOLFIRI) have significantly improved clinical efficacy as related to overall response rates, time to tumor progression, and median overall survival. Capecitabine, an oral fluoropyrimidine, has now been shown in several phase III studies to be as effective as infusional 5-FU when combined with oxaliplatin. During this same time frame, intense efforts have focused on integrating novel biologic agents with cytotoxic chemotherapy regimens. These biologic agents target critical signaling pathways such as the vascular endothelial growth factor (VEGF) and the epidermal growth factor receptor (EGFR). Bevacizumab is a monoclonal antibody targeting VEGF-A, and when combined with fluoropyrimidine based chemotherapy, which includes oxaliplatin or irinotecan, significantly improves clinical efficacy in the management of patients with mCRC. Cetuximab and panitumumab are monoclonal antibodies targeting EGFR, and each of these agents is approved to treat mCRC patients who have progressed on previous chemotherapy treatments. Recent studies have shown that cetuximab, when combined with cytotoxic chemotherapy, significantly enhances the management of patients with mCRC in the first-, second-, and disease-refractory settings. With these advances in treatment options, much attention is now focused on identifying the critical molecular biomarkers that can predict response and/or toxicity to facilitate the evolution of empiric chemotherapy to individually tailored treatments for patients with mCRC. PMID- 17921725 TI - Renal cell cancer. AB - Metastatic renal cell cancer has traditionally been treated with interferon and interleukin-2. An improved understanding of the biology of renal cancer has engendered novel targeted therapeutic agents that have altered the natural history of this disease. The vascular endothelial growth factor and its related receptor and the mammalian target of rapamycin signal transduction pathway in particular have been utilized as therapeutic targets. Sunitinib malate, sorafenib tosylate, temsirolimus, and bevacizumab/interferon alfa have improved clinical outcomes in randomized trials. Other antiangiogenic agents have also demonstrated activity in early studies. Given the availability of multiple treatment options, several questions emerge as to how to integrate these new therapies into the management of metastatic renal cell cancer. Recently reported and planned clinical trials will help clarify the role of these agents. The future of therapy for renal cancer appears promising owing to the efficacy of these novel agents. PMID- 17921727 TI - Active surveillance with selective radical treatment for localized prostate cancer. AB - The challenge of managing localized prostate cancer is to distinguish patients with clinically relevant cancers, who may benefit from radical treatment, from the remainder who do not need any intervention. Active surveillance with selective radial intervention is a management strategy that offers patients the hope of avoiding "unnecessary" treatment without detriment to their long-term survival. Here we discuss the rationale for active surveillance, and the early results. There is no consensus on the optimum active surveillance protocol, with uncertainty regarding the interpretation of PSA kinetics, repeat biopsy results and prostate imaging. In the future, it is likely that molecular markers will revolutionize our ability to select patients who will benefit from definitive treatment. In the meantime, active surveillance provides an attractive way of reducing over treatment. PMID- 17921728 TI - High-dose external beam radiation for localized prostate cancer: current status and future challenges. AB - Since the 1960s, external beam radiation has been one of the major curative treatment options for patients with clinically localized prostate cancer. Efforts to improve the efficacy of this modality have focused on delivering a higher dose, and several recent randomized trials have confirmed that this higher dose results in improved oncological outcomes, particularly for patients with intermediate-risk disease. Technological advancements over the past 2 decades have allowed highly conformal treatments that spare more normal tissue and reduce early and long-term treatment side effects. In a complementary fashion, methods have been developed for better real-time localization of the prostate such that radiation fields can be shifted before each treatment to match the daily shifts in the position of the target, leading to greater accuracy and allowing for smaller treatment margins that in turn will overlap with less normal tissue. With newer and more expensive technologies such as intensity-modulated radiation therapy and protons being used with increasing frequency for the treatment of prostate cancer, it becomes imperative to study the risks and benefits of each new modality so that informed cost-benefit decisions can be made. Similarly, there has been a growing interest in hypofractionation as a means of exploiting the supposed low alpha/beta ratio of prostate cancer to shorten overall treatment time and thereby improve convenience and lower costs. However, as with any new technology, it is necessary to proceed with caution in the arena of hypofractionation while we await the results of trials that will help us to determine the long-term risks and benefits of hypofractionation and whether biological assumptions about the underlying alpha/beta ratio can translate into a true clinical advantage. PMID- 17921729 TI - Brachytherapy for the treatment of prostate cancer. AB - Low-dose rate brachytherapy has become a mainstream treatment option for men diagnosed with prostate cancer because of excellent long-term treatment outcomes in low-, intermediate-, and high-risk patients. Largely due to patient lead advocacy for minimally invasive treatment options, high-quality prostate implants have become widely available in the US, Europe, and Japan. The reason that brachytherapy results are reproducible in several different practice settings is because numerous implant quality factors have been defined over the last 20 years, which can be applied objectively to judge the success of the intervention both during and after the procedure. In addition, recent long-term follow-up studies have clarified that the secondary cancer incidence of brachytherapy is not clinically meaningful. In terms of future directions, the study of radiation repair genetics may allow for the counseling physician to better estimate any given patients risk for side effects, thereby substantially reducing the therapeutic uncertainties faced by patients choosing a prostate cancer intervention. PMID- 17921730 TI - The current use and future trends of focal surgical therapy in the management of localized prostate cancer. AB - The current treatment algorithms for management of localized prostate cancer are mainly extirpative in nature. Treatment varies from expectant management to radical prostatectomy or radiation therapy. However, the ever-increasing emphasis on achieving the best survival benefit while better preserving quality of life, coupled with the introduction of new, safer, and efficacious minimally invasive ablative technologies, has led to the increased popularity of minimally invasive treatment (MIT). MIT refers to the use of a wide range of techniques for local target ablation of the prostate gland with minimal damage to the surrounding tissue. Currently these include cryotherapy and high-intensity focused ultrasound. However, other experimental technologies such as photodynamic therapy, interstitial prostate brachytherapy, and microwave and radiofrequency interstitial tumor ablation are also currently under investigation in early clinical trials. To date, the overall interim results for these relatively new modalities of treatment appear comparable to those for surgical and radiation therapies. However, randomized, controlled studies are needed to support use of these modalities as an alternative to surgery and radiation. In this review, we will address the current rationale for and knowledge of MIT with regard to its safety and efficacy in the treatment of localized prostate cancer. In addition, we will discuss future promising tools in MIT such as photodynamic therapy and the target focal therapy approach as a new trend for the treatment of organ confined low-volume disease. PMID- 17921731 TI - Quality of life after therapy for localized prostate cancer. AB - Given the lack of unequivocal survival data clearly favoring one treatment over another for localized prostate cancer, patients strongly consider quality-of-life effects when choosing treatment for this common malignancy. In the past 15 years, a sizeable body of literature assessing health-related quality-of-life (HRQOL) outcomes in localized prostate cancer has emerged. The goal of this article is to review the quality-of-life experience after treatment for localized prostate cancer. Specifically, I will briefly discuss how quality of life is measured and then review the quality of life effects of each of the commonly used treatment strategies in localized prostate cancer. Finally, I attempt to directly compare the quality-of-life effects of the various treatments to assist clinicians in advising patients with newly diagnosed localized prostate cancer. PMID- 17921735 TI - Aortic valve-sparing operations for aortic root and ascending aortic aneurysms. AB - PURPOSE OF REVIEW: Techniques of aortic root replacement have been developed that preserve the native aortic valve. These techniques avoid anticoagulation in patients who would otherwise receive a composite valve graft with a mechanical valve. RECENT FINDINGS: Longer-term data on the longevity of the main two techniques of aortic valve-sparing operations, the root remodeling and the valve reimplantation technique, are now becoming available. Root remodeling appears to have acceptable outcomes in patients without annuloaortic ectasia or Marfan syndrome. In such patients, the aortic valve reimplantation technique provides superior longevity because it provides external support for the aortic annulus and prevents ongoing dilatation. SUMMARY: Although aortic valve-sparing operations are being performed more frequently, particularly in younger patient populations, these procedures have not been adopted in the current guidelines. The excellent long-term outcomes of such procedures should facilitate their widespread adoption. PMID- 17921733 TI - Mechanisms operative in the antitumor activity of temozolomide in glioblastoma multiforme. AB - PURPOSE: Glioblastoma multiforme (GBM) is the most frequent and incurable brain tumor in adults. Although temozolomide (TMZ) does not cure GBM, it has demonstrated anti-GBM activity and has improved survival (8-14 months) and quality of life. We studied the mechanisms by which TMZ affects 2 human GBM cell lines; U251-MG and U87-MG, aiming to unravel the drug-activated cascades to enable the development of combination therapies that will improve the efficacy of TMZ. MATERIALS AND METHODS: The 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium assay was used to assess cell viability. Modulation of gene expression by TMZ therapy was assayed by gene profiling and verified by quantitative real-time polymerase chain reaction. Protein levels influenced by the treatment were studied by Western blots and immunocytochemistry. RESULTS: Increasing concentrations of TMZ decreased cell viability in a concentration dependent manner. The expression of 1,886 genes was altered >2-fold after TMZ treatment. We focused on the 81 genes similarly altered by TMZ treatment in both cell lines to neutralize tissue-specific characteristics. Fourteen target genes of hypoxia-inducible factor (HIF-1), were found to be up-regulated after TMZ treatment including vascular endothelial growth factor (VEGF). HIF-1alpha expression was constant at the mRNA level; however, its post-treatment protein levels increased compared with those of untreated control cells. DISCUSSION: The genetic analyses suggest that treatment with TMZ activates stress mechanisms in GBM cells that include the angiogenesis-inducing proteins HIF-1alpha and VEGF. We propose that treatment with TMZ be supplemented with either an antibody to VEGF or down-regulators of HIF-1alpha to improve clinical results of TMZ in the treatment of GBM. PMID- 17921736 TI - Intramural hematoma and penetrating aortic ulcer. AB - PURPOSE OF REVIEW: With advances in imaging technology, increased attention has turned to the 'variant forms' of aortic dissection: intramural hematoma and penetrating aortic ulcer. At the same time, the advent of endovascular therapies such as stent-grafting has broadened the base of practitioners capable of intervening in these pathologies from the cardiovascular surgeon to now include vascular surgeons, interventional radiologists, and invasive cardiologists. Accordingly, a reassessment of these conditions is of general interest. RECENT FINDINGS: The natural history of both entities is being elucidated with increasing precision as relatively large single-center and multicenter studies are being reported. Recent attention has focused on application of endovascular technologies to penetrating aortic ulcer, while the principal controversy over intramural hematoma concerns its management when the ascending aorta is involved. SUMMARY: Despite continuing controversy over the outcome of penetrating aortic ulcer managed by medical or open surgical means, thoracic endograft technology is being applied to this entity with high procedural success and low perioperative morbidity by experienced teams internationally. The benefit to patients as the use of this technology expands, however, will depend critically upon selection criteria. Evidence in favor of surgical management of type A intramural hematoma continues to mount, although it is also clear that, in specific circumstances, a nonoperative approach may suffice. PMID- 17921732 TI - CEOP-21 versus CEOP-14 chemotherapy with or without rituximab for the first-line treatment of patients with aggressive lymphomas: results of the HE22A99 trial of the Hellenic Cooperative Oncology Group. AB - BACKGROUND: In this study we investigated whether administering CEOP (cyclophosphamide, epirubicin, vincristine [Oncovin], and prednisone) every 2 weeks (CEOP-14) instead of every 3 weeks (the standard CEOP-21 regimen) improves outcomes in patients with previously untreated aggressive lymphomas. In a secondary analysis we evaluated the impact of adding rituximab to CEOP-14/CEOP-21 chemotherapy. STUDY DESIGN: The trial opened in March 1999, and patients were randomly assigned to either CEOP-14 or CEOP-21. All patients enrolled from May 2002 onward received rituximab with each chemotherapy cycle, and those attaining a complete response received rituximab consolidation. RESULTS: Complete and overall response rates in the CEOP-21 +/- rituximab (N = 114) and CEOP-14 +/- rituximab (N = 103) arms were similar, as were the overall survival (P = 0.769) and time to progression distributions (P = 0.969). Rituximab was shown to have a beneficial effect both on the overall survival and on the time to progression. CONCLUSIONS: Thus far, no significant improvement in outcome has been demonstrated with CEOP-14 +/- rituximab versus CEOP-21 +/- rituximab. However, with addition of rituximab to CEOP-21/CEOP-14, significant improvements in time to progression and overall survival were achieved. PMID- 17921737 TI - Stent graft treatment for transverse arch and descending thoracic aorta aneurysms. AB - PURPOSE OF REVIEW: The aim of this article is to review the recent relevant literature on endovascular repair of thoracic aortic aneurismal disease. RECENT FINDINGS: The introduction of endovascular stent graft technology has ushered in a new era in therapy for diseases of the aortic arch and descending thoracic aorta. The technical challenges of stent graft deployment in the descending thoracic aorta, such as proximity to the great vessels and arch tortuosity, have been and remain a device engineering focus. More recently, repair of aortic arch aneurysms has been accomplished using both 'hybrid' (open and endovascular) and totally endovascular techniques. SUMMARY: Endovascular stent grafting of aneurismal disease processes of the thoracic aorta is feasible and relatively safe. Exquisite judgment is essential for good results. These results generally rest on a broad knowledge base of thoracic aortic disease processes and experience in both open and endovascular surgery. Careful attention to patient anatomy and device specifications must be maintained. The key to the successful implementation of this technology lies in careful preoperative planning, intraoperative execution with safe device delivery, and prevention of central nervous system injury. Routine follow-up imaging is imperative to better understand the long-term results and indications for these new procedures. PMID- 17921738 TI - Coronary artery bypass graft vs. percutaneous coronary angioplasty: CABG on the rebound? AB - PURPOSE OF REVIEW: To examine the benefits of coronary artery bypass grafting (CABG) against percutaneous coronary intervention (PCI) and particularly the use of drug-eluting stents (DES) in situations where CABG has traditionally been considered the most effective therapy on clinical and economic grounds. RECENT FINDINGS: Current studies reconfirm that CABG is still the best therapy in terms of improved survival and freedom from reintervention for most patients with proximal left anterior descending, multivessel and left main-stem coronary artery disease (CAD) and that these benefits are even greater in diabetic patients. Health economic analyses also confirm the cost-effectiveness of medical therapy and CABG but not PCI. Furthermore, several meta-analyses have shown that DES do not improve survival or freedom from myocardial infarction compared with bare metal stents, but increase the risk of stent thrombosis, with associated medical and financial implications for prolonged dual antiplatelet medication. SUMMARY: In view of the evidence in favour of CABG, this article questions the justifiability of some trials of PCI vs. CABG, especially in diabetic patients and those with left main-stem CAD, and exhorts the need for a multidisciplinary team approach to the management of CAD as a minimum standard of care. PMID- 17921739 TI - Clinical trials versus registries in coronary revascularization: which are more relevant? AB - PURPOSE OF REVIEW: Clinical decision-making in coronary artery disease relies heavily on evidence-based medicine. Data from randomized controlled trials constitute the highest order of evidence and remain the standard for comparisons between therapies. While comprehensive, observational databases lack the scientific rigor of randomized controlled trials, they represent a more accurate accounting of everyday clinical care. Which data are more relevant to clinical practice?. RECENT FINDINGS: At least 11 randomized controlled trials and three meta-analyses comparing coronary artery bypass grafting and percutaneous coronary intervention exist, which all largely show no difference in death or myocardial infarction between the two treatments but more repeat revascularization with percutaneous coronary intervention. All these studies, however, are subject to the biases of trial design, which impact the external validity of the results. Analyses of four observational databases show a survival advantage in multivessel disease with coronary artery bypass grafting. Although these are reflective of real world clinical practice, they are subject to 'treatment bias', some of which can be corrected by risk adjustment. SUMMARY: Information from both randomized controlled trials and outcomes databases is necessary to determine appropriate strategy for individual patients. Reliance on data solely from either source is insufficient. It is incumbent on the treating physician to know not only the results of published studies, but also the limitations of that information. PMID- 17921740 TI - Multiple extensive coronary artery stenting: does it compromise future surgical revascularization? AB - PURPOSE OF REVIEW: In the era of percutaneous coronary intervention, surgeons are confronted with performing coronary artery bypass graft surgery on patients with previous balloon dilatation or stenting. This review evaluates the impact of previous percutaneous coronary intervention on patient survival and choice of optimal myocardial revascularization technique. RECENT FINDINGS: Aggressive atherosclerosis has been remarked in patients complicated with intrastent stenosis. Moreover, bypass grafting with venous grafts has shown an extremely high incidence of graft failure in the restenosis population due to limited nitric oxide (a natural vasodilator) production of venous grafts. The challenge is to achieve complete revascularization in an unfavourable setting (greater co morbidities, complex coronary lesions) with a greater risk of graft occlusion. SUMMARY: The internal thoracic artery is the optimal graft for myocardial revascularization in patients with and without previous in-stent restenosis. Coronary artery reconstruction by exclusive internal thoracic artery grafting gives superior patency rates and clinical outcomes. It is the most appropriate approach for myocardial revascularization in these patients. PMID- 17921741 TI - Radial artery conduit for coronary revascularization: as good as an internal thoracic artery? AB - PURPOSE OF REVIEW: Multiple arterial grafting has been proposed to improve outcomes after coronary bypass surgery. We sought to define the emerging role of the radial artery as an alternative second arterial graft. RECENT FINDINGS: Despite significant evidence suggesting superiority of bilateral internal thoracic artery grafting, utilization in current clinical practice remains low due to concerns about sternal wound infection and conduit length. Recent clinical trials have started to clarify the role of the radial artery in coronary surgery. This conduit provides superior patency to saphenous vein grafts in the setting of high-grade stenosis regardless of the target territory and with minimal harvest site complications. SUMMARY: The radial artery is a versatile conduit that provides clinical benefit in a large variety of patients undergoing coronary bypass surgery. Outcomes from ongoing longitudinal clinical trials will further clarify optimal conduit selection strategies. PMID- 17921742 TI - Robotically assisted coronary surgery: what is the future? AB - PURPOSE OF REVIEW: The purpose of this review is to assess the current state of robotics in coronary revascularization and to consider what advances may be forthcoming. RECENT FINDINGS: Several groups have been able to utilize robotic telemanipulation to perform coronary revascularization via limited access. Although the patients are highly selected, results in experienced centers are comparable to open coronary artery bypass grafting. SUMMARY: The ultimate goal of minimally invasive coronary surgery is totally endoscopic coronary revascularization without utilization of cardiopulmonary bypass. This goal can be realized with the utilization of robotic technology. PMID- 17921743 TI - The molecular basis of vulnerable plaque: potential therapeutic role for immunomodulation. AB - PURPOSE OF REVIEW: Atherosclerosis is a chronic inflammatory/immune disease involving multiple cell types including monocytes-macrophages, T-lymphocytes, mast cells, and endothelial cells. Through recent studies the role of the immune system on development of atherosclerosis and approaches to modulate this response are being elucidated. RECENT FINDINGS: The use of statins, PPARgamma agonists or lipoprotein-associated phospholipase A2 (Lp-PLA2) inhibitors may play a role in reducing progression of atherosclerosis through immunomodulatory pathways. Oxidized LDL biases development toward the pro-inflammatory T-cell Th1 subset and recruits macrophages into the vascular wall. IFNgamma, produced by Th1 cells, inhibits PPARgamma effects. Lp-PLA2 levels correlate with an increased risk of recurrent ischemic events in patients presenting with acute coronary syndromes or myocardial infarction. SUMMARY: Recent research has shown that immune pathways play a major role in the development and progression of atherosclerosis. Commonly used medications, specifically statins and some PPARgamma agonists, have demonstrated anti-inflammatory/immune effects unrelated to their primary mode of action. Treatment of infectious agents has proven elusive in the clinical arena. Novel agents targeting immune and inflammatory pathways may prove beneficial in reducing progression and instability of the atherosclerotic plaque. PMID- 17921744 TI - Risk stratification of individual coronary lesions using local endothelial shear stress: a new paradigm for managing coronary artery disease. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize the role of endothelial shear stress in the natural history of coronary atherosclerosis, and to propose an individualized risk-stratification strategy of atherosclerotic lesions based on the in-vivo characterization of local endothelial shear stress and wall morphology. RECENT FINDINGS: Low endothelial shear stress promotes the development of early fibroatheromas, which subsequently follow an individualized natural history of progression. This individual natural history is critically dependent on the magnitude of low endothelial shear stress, which subsequently regulates the severity of inflammation within the wall and ultimately the vascular remodeling response. Very low endothelial shear stress enhances plaque inflammation, leading to excessive expansive remodeling. Excessive expansive remodeling leads to perpetuation, or even exacerbation, of the local low endothelial shear stress environment, thereby setting up a self-perpetuating vicious cycle among low local endothelial shear stress, inflammation, and excessive expansive remodeling, which transforms an early fibroatheroma to a high risk plaque. SUMMARY: In-vivo assessment of the local endothelial shear stress environment, severity of inflammation and vascular remodeling response, all responsible for individual plaque behavior and natural history, in combination with systemic biomarkers of vulnerability, may allow for detailed risk stratification of individual early atherosclerotic plaques, thereby guiding both systemic and local, lesion-specific therapeutic strategies. PMID- 17921745 TI - The conundrum of late and very late stent thrombosis following drug-eluting stent implantation. AB - PURPOSE OF REVIEW: Drug-eluting stents reduce restenosis compared with bare metal stents, but there is growing concern that drug-eluting stents may lead to higher rates of late stent thrombosis, a rare and potentially catastrophic complication following stenting. RECENT FINDINGS: While the data on the risk of late stent thrombosis are not definitive, several general conclusions may be drawn from the available data. Late thrombosis, while associated with high mortality and morbidity, is an uncommon complication of both drug-eluting stents and bare metal stents. Randomized trials of approved drug-eluting stents versus bare metal stents have shown additional cases of late stent thrombosis in drug-eluting stents, but no significant difference in the cumulative incidence of stent thrombosis, myocardial infarction, or cardiac death at 4 years of follow-up. Observational studies suggest higher very late stent thrombosis incidence, but the relative risks of drug-eluting stents versus bare metal stents in specific high-risk groups require further study. Although the etiology of late stent thrombosis is multifactorial, premature discontinuation of clopidogrel appears to be the most important risk factor. SUMMARY: Long-term follow-up of patients after coronary stenting has identified stent thrombosis as a rare but serious event. Ongoing clinical trials in broader patient populations will be helpful to understand the risk of late stent thrombosis with greater certainty. PMID- 17921746 TI - New understanding about calcific aortic stenosis and opportunities for pharmacologic intervention. AB - PURPOSE OF REVIEW: This review article will discuss aortic stenosis, the evolving studies defining the cellular mechanisms and the potential for medical therapies for the treatment of this disease. RECENT FINDINGS: Currently, the only therapy for these patients is surgical valve replacement. In the past decade there has been a change in the paradigm towards our understanding of the cellular biology of this disease process. Studies in laboratories across the world have demonstrated that this disease has an active biology and that this biology may be targeted with medical therapies similar to that of vascular atherosclerosis. SUMMARY: Calcific aortic stenosis is the third most common form of cardiovascular disease in the USA. It has replaced rheumatic heart disease in prevalence in western countries due to improved access to healthcare and the widespread use of antibiotics. PMID- 17921747 TI - Current understanding of heart failure with preserved ejection fraction. AB - PURPOSE OF REVIEW: A substantial proportion of patients with heart failure have preserved ejection fraction. Though patients with preserved ejection fraction experience a substantial burden of morbidity and mortality, the understanding of heart failure pathophysiology in this group remains incomplete and evidence-based therapeutic options are limited. RECENT FINDINGS: The prevalence of heart failure in patients with preserved ejection fraction is increasing and prognosis in this population remains poor despite modern medical therapy. Though diastolic dysfunction is typically present, increasing evidence suggests that extracardiac factors such as renal dysfunction and enhanced central aortic stiffness may play an important role in the development and progression of heart failure symptoms. Results of the first randomized, controlled clinical trials in this population suggest a possible therapeutic role for renin-angiotensin system blockade in reducing heart failure-associated morbidity, but there is still no evidence supported strategy for reducing mortality in this population. SUMMARY: Though the epidemiology and impact of heart failure with preserved ejection fraction are increasingly clear, consensus regarding pathophysiology and the optimal therapeutic approach is still lacking. Pending completion of additional therapeutic trials in this population, treatment remains largely empiric and focused on optimizing myocardial performance in diastole by control of blood pressure, restoration or maintenance of sinus rhythm, and relief of volume overload. PMID- 17921750 TI - Publication of the International Psychogeriatric Association consensus statement on defining and measuring treatment benefits in dementia. PMID- 17921751 TI - Psychogeriatric services in the community and in long-term care facilities: needs and developments. AB - PURPOSE OF REVIEW: An increasing proportion of the world's population is over the age of 65 years. Specialist mental health services for older people have been developed in many countries. The way services develop depends partly on how healthcare arrangements have evolved in that jurisdiction, as well as on finances, culture and attitudes towards elderly and disabled individuals. Health planners in developing countries recognize that considerable increases in their elderly populations and numbers of individuals with disability are imminent. It will be important to ensure that older people with mental disorders receive appropriate treatment. This review discusses recent findings and observations about psychogeriatric services in the community and in long-term care facilities, and aims to suggest how to improve or develop such services. RECENT FINDINGS: The prevalence of mental disorders in long-term care facilities is high, but services to deal with them are usually not optimal. When appropriately staffed and organized, community psychogeriatric services, day care and collaborative care can be effective in reducing mental health problems and preventing admissions. SUMMARY: Recent reviews and research have provided useful guidance regarding aspects of current psychogeriatric services that work well and those that need to be improved. A person-centred approach is favoured. PMID- 17921752 TI - Recent understandings in geriatric affective disorder. AB - PURPOSE OF REVIEW: This article focuses on recent research into depression, bipolar disorder and anxiety in older people. RECENT FINDINGS: Many physical illnesses are associated with a high prevalence of depression but overall medical burden may largely account for this. The relationship between depression and vascular disease is two way. Frontal brain dysfunction may underlie depression both in cerebrovascular disease and neurodegenerative disorders. Besides antidepressants, psychological treatments, psychosocial interventions and enhanced primary care services are effective. Longer-term outcomes are poor but preventive strategies show promise. Medical and psychiatric comorbidity are also important themes in later-life anxiety and bipolar disorders. SUMMARY: Improving prognosis is a key concern and more research into novel pharmacological approaches (including vasoprotection), psychological interventions and prevention is needed. PMID- 17921753 TI - Studies from emerging countries: an encouraging development. AB - PURPOSE OF REVIEW: To summarize the findings of recent psychogeriatric research conducted in emerging countries. RECENT FINDINGS: During the review period there have been new reports on the prevalence, clinical characteristics and pathogenic mechanisms of dementia in its various forms, epidemiological findings on depression, and psychological autopsy analyses of suicidal behaviors and psychiatric morbidities. New studies in social psychiatry have been conducted, especially on the trends of transgenerational support of the elderly and of interventions that may reduce psychiatric disease burdens and promote mental health in later life. SUMMARY: These research findings support a better understanding of the prevalence, nature, and underlying risk and protective factors of the major psychiatric morbidities in the studied regions. Further exploration of the epidemiological profile of mental disorders in rural areas, research on the direct and indirect costs of psychogeriatric conditions, and evaluation of the cost-effectiveness and financial sustainability of early detection programs, treatment and rehabilitation models are warranted. PMID- 17921754 TI - The many faces of psychosis in the elderly. AB - PURPOSE OF REVIEW: As the population ages, the number of older patients with psychosis will greatly rise. This review focuses on the etiology, biologic and clinical findings, and treatments of common causes of psychosis in the elderly. RECENT FINDINGS: Recent studies on psychosis related to Alzheimer's disease indicate that antipsychotic drugs have equivocal efficacy in improving psychotic symptoms and may have side effects or risks that outweigh their benefits. Behavioral interventions for agitation in dementia are showing some promise. In older adults with schizophrenia, intramuscular ziprasidone was found to be effective, and evidence is emerging for the use of hormone replacement therapy. For depression with psychosis, a recent study found that the combination of an antidepressant with an antipsychotic is no more effective than an antidepressant alone. SUMMARY: There is support for the use of antipsychotic drugs for all types of psychosis in the elderly. While the atypical antipsychotics have a 'black box warning' on risk of death in elderly patients with dementia, the typical antipsychotics carry an even higher risk of death and adverse effects. Weighing the potential risks and benefits of treatment options is essential. Please refer to your country's regulations regarding the use of antipsychotic drugs. PMID- 17921755 TI - Missing links in phenomenological clinical neuroscience: why we still are not there yet. AB - PURPOSE OF REVIEW: The phenomenology or systematic study of the patient's subjective experience in neuropsychiatric disorders is widely recognized as important. The methods used, the type of 'knowledge' obtained and the relationship of these observations to standard methods of clinical neuroscience, however, remain ill-defined and highly controversial. RECENT FINDINGS: Advances in the phenomenology of consciousness, self, body-experience, time-perception and intersubjectivity of neuropsychiatric disorders have been made. This review examines two differing approaches to the phenomenological psychiatry-neuroscience interface: the neo-phenomenological approach claims that some of its key concepts (e.g. the hyperreflexivity/ipseity model, prereflective self-awareness) are able to constrain neuroscience; the existential-phenomenological approach counters that phenomenology's role is not to constrain neuroscience but to provide hypotheses for further experimental study. This review compares these two approaches and assesses the current success of their respective claims. SUMMARY: By integrating work of largely untranslated authors, such as Binswanger, Blankenburg, and von Weizsaecker, neglected or cited out of context in the neo phenomenological approach, the existential-phenomenological approach provides the 'missing links' between phenomenology and clinical neuroscience in a newly emerging but still fragile balance between disciplines. Optimistic claims about the ability of recently proposed phenomenological concepts (neo-phenomenological approach) to constrain neuroscience are unwarranted. PMID- 17921756 TI - Diversity. PMID- 17921757 TI - New directions in the conceptualization of psychotic disorders. AB - PURPOSE OF REVIEW: Recent studies raise controversies about the nature of psychotic illnesses, and the role of life experiences and drug abuse as causative agents in the onset of psychoses. RECENT FINDINGS: Evidence from studies across many geographic locales and cultures finds increased risk of psychoses in first- and second-generation immigrant populations. Trauma incurred in war and civil unrest, trauma of child abuse, and the experience of being bullied in childhood are correlated with increased rates of psychoses in the populations at risk. The risk of onset of psychoses is increased by maternal and infant starvation, and by substance misuse (marijuana, khat) in late childhood and adolescence. These studies question the validity of a categorical distinction between the schizophrenic and affective illnesses. SUMMARY: A variety of extrinsic factors, such as in-utero and infant malnutrition, substance abuse, and traumatic experiences, appear to be significant risk factors for the development of schizophrenia-like and psychotic affective disorders. These findings raise the issue of whether the present classification of the psychoses is in urgent need of reconceptualization. PMID- 17921758 TI - Ethical and conceptual aspects of capacity assessments in psychiatry. AB - PURPOSE OF REVIEW: To search the literature on conceptual and assessment issues of patient capacity. RECENT FINDINGS: Current literature shows that many instruments have been developed in the last decade to measure patient capacity. Although these measures provide a rank-ordered scale of capacity, they cannot categorize patients into competent and incompetent, which relies heavily on the concept of authentic autonomy. The latter, however, should be carefully examined after considering the patient's cultural and subcultural background, and the quality of the doctor's communication skills. SUMMARY: Academic and clinical psychiatry are posed with such questions as to how to categorically classify capacity and incapacity as well as to evidence the admissibility of measuring instruments when used in a civil commitment. PMID- 17921759 TI - Criterion B (social dysfunction) in persons with schizophrenia: the puzzle. AB - PURPOSE OF REVIEW: This review focuses on social dysfunction in persons with schizophrenia and addresses three main questions. What is the core ability that is disordered in social dysfunction according to mainstream paradigms? How is social dysfunction primarily assessed in current research? Which levels of personal experience, and which trans-personal and sub-personal factors, are assessed by mainstream empirical research? RECENT FINDINGS: Empirical studies on social dysfunction include research on trans-personal factors (stigma, availability of social and psychiatric facilities, and family resources), sub personal factors (neurocognition) and personal factors (social cognition, coping and noncognitive factors). The main recent findings of these approaches are described and commented upon. Inconsistencies between studies are identified. SUMMARY: What is it like to be a person with schizophrenia in the social world? Building upon empirical research, we can finally address this question. We argue that more qualitative research is needed into the reasons that persons with schizophrenia have for adopting or embracing their given type of relatedness to the others. PMID- 17921760 TI - Mental health advocacy with black and minority ethnic communities: conceptual and ethical implications. AB - PURPOSE OF REVIEW: Recent policy and legislative developments confirm the right to advocacy for people experiencing mental health problems. Provision of mental health advocacy for black and minority ethnic communities is particularly identified as a key objective for Primary Care Trusts. This review considers the implications for commissioners and practitioners of ensuring access to appropriate advocacy for service users from black and minority ethnic community. RECENT FINDINGS: There is a dearth of literature on mental health advocacy with black and minority ethnic communities. The available research suggests that mainstream conceptualizations of advocacy are not meaningful to these communities and this adversely affects access. Advocacy provided by black community and voluntary sector organizations begins with understanding racism and social disadvantage. It is therefore concerned with addressing these inequalities and resonates strongly with current concepts of recovery. SUMMARY: People from black and minority ethnic communities are not well served by current provision of mental health advocacy, and there is a risk that this will be replicated in the development of independent mental health advocacy, under the amendments to the UK Mental Health Act of 1983. This largely reflects the conceptualization of advocacy and its purpose. The implications for both commissioning and provision of advocacy are explored. PMID- 17921761 TI - Spirituality and mental health: a very preliminary overview. AB - PURPOSE OF REVIEW: Demand for the spiritual dimension to be taken account of in the diagnosis, treatment and care of people with mental ill-health has come strongly from users, carers and professionals. RECENT FINDINGS: Research in the US over the past 10 years has shown a clear correlation between affiliation to a religious group and better outcomes in terms of mental and physical health, and even longevity. In the UK, however, the evidence from research is much less clear, and needs a more focused approach to really elucidate what helps people stay mentally healthy and recover from mental ill-health. SUMMARY: Spirituality is now a key issue as individuals, communities and mental health services struggle to combine technical efficacy and business efficiency, but remain human, so as to nurture service users, carers and staff. This is set in a postmodern world, scarred by the trauma of 9/11 and its aftermaths, and in the context of a global consumerism, which has resulted in individuals being increasingly atomized and isolated. A consumerist society means that those classed as 'deficient consumers', especially those whose ill-health and/or poverty excludes them from the marketplace, are seen as outsiders and a dangerous class. PMID- 17921762 TI - History of psychopathology, psychiatric treatments and the psychiatric profession. AB - PURPOSE OF REVIEW: This review addresses articles published on the subject of the history of psychiatry during 2006 and 2007. RECENT FINDINGS: Original findings in the fields of psychopathology, treatments, the psychiatric profession, its institutions and psychiatrists are described. SUMMARY: Research into psychopathology continues to shed light on how modern society can learn from the past in terms of issues such as whether people are ill, whether they are responsible for their actions, and what constitutes an effective and ethical treatment. PMID- 17921763 TI - Recent translations of Foucault on mental health. AB - PURPOSE OF REVIEW: The work of the French philosopher and historian, Michel Foucault, often dealt with subjects either directly or indirectly related to psychiatry. In the past, his work has been largely ignored or rejected by mainstream psychiatry. RECENT FINDINGS: In the period under review, two important English translations of Foucault's work on psychiatry have been published. Our review focuses on these books, and also looks at some of the recent secondary literature relating to Foucault and mental health. SUMMARY: We argue that psychiatry has a lot to gain from a positive engagement with Foucault's ideas. PMID- 17921764 TI - Pharmacological treatments for paraphilic patients and sexual offenders. AB - PURPOSE OF REVIEW: This review addresses testosterone-lowering and other psychotropic medications for the treatment of paraphilic patients or sexual offenders. RECENT FINDINGS: Randomized controlled studies are still lacking, and only a few new studies were reported during the past year. On the other hand, there is substantial scientific knowledge about the wide range of psychiatric comorbidity associated with paraphilias and in sexual offenders. Empirically based treatment of these patients, especially of impulsivity, anxiety and mood disorders, may also ameliorate sexual impulsivity. SUMMARY: Medication interventions, either substantially lowering serum testosterone or treating axis I comorbidities, show definite promise as a significant component of the management of sexual offenders. Pharmacotherapy should be combined with other therapeutic treatment modalities, most commonly cognitive-behaviour based psychotherapy and intensive community supervision. PMID- 17921765 TI - Clinical challenges in the treatment of patients with posttraumatic stress disorder and substance abuse. AB - PURPOSE OF REVIEW: The aim of this article is to review the current literature on co-occuring posttraumatic stress disorder and substance-use disorder, with an emphasis on clinical aspects and emerging treatments. RECENT FINDINGS: In clinical populations (focusing on either disorder), about 25-50% have a lifetime dual diagnosis of posttraumatic stress disorder and substance-use disorder. Patients with both disorders have a more severe clinical profile than those with either disorder alone, lower functioning, poorer well being, and worse outcomes across a variety of measures. In recent years, several promising treatment programs have been developed specifically for co-occuring posttraumatic stress disorder and substance-use disorder, with one model having been established as effective thus far. SUMMARY: Comorbid posttraumatic stress disorder/substance-use disorder is a frequent diagnosis in clinical populations that severely affects course and outcome. Treatment approaches appropriate for this vulnerable population need to be evaluated further and implemented in routine practice. PMID- 17921766 TI - Metacognitive training in schizophrenia: from basic research to knowledge translation and intervention. AB - PURPOSE OF REVIEW: There has been a marked increase in the study of cognitive biases in schizophrenia, which has in part been stimulated by encouraging results with cognitive-behavioral interventions in the disorder. We summarize new evidence on cognitive biases thought to trigger or maintain positive symptoms in schizophrenia and present a new therapeutic intervention. RECENT FINDINGS: Recent studies indicate that patients with paranoid schizophrenia jump to conclusions, show attributional biases, share a bias against disconfirmatory evidence, are overconfident in errors, and display problems with theory of mind. Many of these biases precede the psychotic episode and may represent cognitive traits. Building upon this literature, we developed a metacognitive training program that aims to convey scientific knowledge on cognitive biases to patients and provides corrective experiences in an engaging and supportive manner. Two new studies provide preliminary evidence for the feasibility and efficacy of this approach. SUMMARY: The gap between our advanced understanding of cognitive processes in schizophrenia and its application in clinical treatment is increasingly being narrowed. Despite emerging evidence for the feasibility and efficacy of metacognitive training as a stand-alone program, its most powerful application may be in combination with individual cognitive-behavioral therapy. PMID- 17921767 TI - What do general medical guidelines say about depression care? Depression treatment recommendations in general medical practice guidelines. AB - PURPOSE OF REVIEW: Depression is one of the most costly and prevalent mental disorders, and it often co-occurs in patients with general medical conditions. This review identifies the extent to which depression treatment is being addressed in guidelines for general medical illness treatment. RECENT FINDINGS: We conducted a review of the literature published within the past year pertaining to depression treatment guidelines in medical care. We also conducted a systematic review of available practice guidelines listed by the Agency for Research and Quality to assess the extent to which depression treatment is being addressed in current general medical treatment guidelines. Overall, only a handful of articles addressed treatment of depression within the context of general medical care, and most only addressed screening. Among 199 available practice guidelines for general medical conditions, only 83 mentioned depression treatment, and the majority of these only addressed depression screening. SUMMARY: To improve outcomes in patients with co-occurring general medical conditions, general medical guidelines should include specific recommendations for treating and co-managing depression. Additional strategies that facilitate the translation of guidelines into routine practice, address the management of multiple chronic illnesses, and reduce the stigma of mental illness are also needed. PMID- 17921768 TI - Bibliography. Current world literature. PMID- 17921770 TI - Reconstruction in urology. PMID- 17921771 TI - Patterns of urethral injury and immediate management. AB - PURPOSE OF REVIEW: The management of urethral trauma remains controversial. The purpose of this review is to provide a concise account of the patterns of urethral injury and the current trends of its immediate management. RECENT FINDINGS: Recent studies have identified types of pelvic fractures associated with a higher risk of urethral injury. Recognition of these fractures coupled with timely radiographic imaging has facilitated the earlier diagnosis of urethral injury and ensured that serious long-term sequelae are minimized. In highly selected cases of complete anterior and posterior urethral disruption primary realignment by a combined antegrade and retrograde endoscopic approach is increasingly being considered as a management option to restore early urethral continuity. Urethral trauma secondary to penetrating gunshot wounds and penile fracture requires immediate surgical exploration and repair. SUMMARY: The urologist involved in the management of genitourinary tract trauma needs to recognize the patterns of urethral injury, especially those associated with certain pelvic fractures. A judicious and systematic approach coupled with a conscious effort to minimize short and long-term sequelae of all urethral injuries will ensure optimal results. There is a need for a consensus on the optimal management of each of the vast arrays of urethral injuries. PMID- 17921772 TI - Reconstruction of the lower urinary tract by laparoscopic and robotic surgery. AB - PURPOSE OF REVIEW: Minimally invasive techniques are being developed at a rapid rate, although not all of these stand up to close peer-review and scrutiny. We describe the current state of laparoscopy and robotic-assisted reconstructive urological surgery on the lower urinary tract. These procedures are technically demanding and require advanced laparoscopic skills, including suturing. RECENT FINDINGS: Techniques for urethra-vesical anastomosis following radical prostatectomy and reconstruction after radical cystectomy are discussed. In addition, minimally invasive techniques for bladder augmentation, colposuspension and ureteric reimplantation are reviewed. SUMMARY: We indicate both the reconstructive procedures supported by sound evidence and those with little hard data backing them up. PMID- 17921773 TI - Tissue glues and nonsuturing techniques. AB - PURPOSE OF REVIEW: This article details the diverse urologic applications of tissue glues and hemostatic agents over the past 3 years in the management of genitourinary injuries, surgical wounds, and complications. RECENT FINDINGS: Biosurgical agents designed to promote tissue adhesion and hemostasis are being increasingly employed across all surgical disciplines. Fibrin sealant is the most widely utilized biosurgical product. Gelatin matrix thrombin has proven to be an efficacious hemostatic agent. Bovine serum albumin-gluataraldehyde is a new, promising tissue glue. Complex reconstructive, oncologic and laparoscopic procedures are those most appropriate for sealant use in urology. SUMMARY: Tissue glues and hemostatic agents are effective, safe, and their use is increasing. All urologists should have a working knowledge of these adjuncts. PMID- 17921774 TI - Is there any evidence that one continent diversion is any better than any other or than ileal conduit? AB - PURPOSE OF REVIEW: There is an obvious discrepancy between the perceived advantages of the different forms of continent urinary diversion from an expert's point of view and the limited utilization of these techniques outside of academic and tertiary referral centers. RECENT FINDINGS: The vast majority of studies are retrospective with numerous confounding factors and often poorly defined outcomes. The rates of postoperative morbidity and need for reoperation varies widely among studies, even for the same procedure. The utilization of continent diversion depends on demographic, socioeconomic, provider-based, and clinical variables, with low case volume and lack of experience probably being the most obvious reasons for avoidance of more complex techniques. In contrast to the prevailing notion that patients undergoing continent diversion have superior quality of life than those receiving a conduit, existing studies fail to show significant advantages of one technique over the other. SUMMARY: The current body of published literature is insufficient to conclude that there is a superior form of urinary diversion in terms of evidence-based medicine. It is quite clear, however, that not all patients are candidates for one type of diversion. The best results are obtained when a comprehensive concept is tailored to the individual patient. PMID- 17921775 TI - The use of biological and synthetic materials in vaginal surgery for prolapse. AB - PURPOSE OF REVIEW: This article reviews the currently available biological and synthetic graft materials available for pelvic prolapse. It also focuses on recent publications examining their use in transvaginal prolapse surgery. RECENT FINDINGS: There continue to be many case series investigating the treatment of anterior prolapse, however there are still very few publications examining new techniques involving apical fixation with large anterior/posterior meshes. Of note are three randomized studies that have recently been reported--two with anterior prolapse, one showing a lack of significant benefit of cadaveric fascia, one demonstrating the benefit of porcine dermis, and one with posterior prolapse demonstrating a lack of benefit of porcine SIS. SUMMARY: Despite the paucity of randomized control trials of various graft materials used in transvaginal prolapse surgery, the many case series published to date generally show relative safety and efficacy. There is still a need for more randomized controlled trials, however, to guide therapy and for well designed case series with longer follow up, especially regarding new techniques involving large anterior/posterior meshes. PMID- 17921776 TI - Vaginoplasty. AB - PURPOSE OF REVIEW: This review summarizes the most recent advances in the field of vaginoplasty. New surgical techniques and recent modifications to existing techniques are included as well as up-to-date outcome data when available. RECENT FINDINGS: It is increasingly clear that vaginal dilation treatment is a successful method of vaginal creation and avoids the risks of surgery. When vaginal dilation is unsuitable or unsuccessful, however, vaginal reconstruction surgery is needed. Most of the novel advances in vaginal reconstruction have been related to the advances made in laparoscopic techniques. It is anticipated that as surgeons become more adept at such procedures, maximally invasive procedures such as intestinal neovagina may be necessary only in a small minority of patients. Autologous vaginal tissue transplantation may also transform this field. Long-term sexual function outcome data are essential for full evaluation and comparison of surgical techniques and the recent literature has begun to include this information. SUMMARY: Vaginal dilation treatment should be the first choice of treatment. If unsuccessful, laparoscopic techniques offer good outcomes and quick recovery. Additional research into the long-term outcomes of sexual satisfaction, orgasm and patient body image is essential, however, to allow a comparison of newer and traditional procedures. PMID- 17921777 TI - How worrisome is testicular microlithiasis? AB - PURPOSE OF REVIEW: Anecdotal reports of men developing testicular cancer after previous identification of microcalcifications on ultrasound generated significant concern in the literature about the relationship of testicular microlithiasis and testis cancer. Until 2001 little prospective data were available on the prevalence or natural history of testicular microlithiasis in the healthy male population. RECENT FINDINGS: Testicular microlithiasis is present in 5.6% of the male population between 17 and 35 years of age (14.1% in African Americans), far more common than testicular cancer (7:100,000). The majority of men with testicular microlithiasis will not develop testicular cancer. Previously recommended surveillance regimens using ultrasound, tumor markers or testicular biopsy are too costly and do not offer an improved outcome over testicular self-examination. SUMMARY: Testicular microlithiasis is common and while microcalcifications do exist in roughly 50% of germ cell tumors the majority of men with testicular microlithiasis will not develop testicular cancer. Increased emphasis on testicular examination is the recommended follow up for men identified with testicular microlithiasis. PMID- 17921778 TI - Pathologic effects of testosterone deprivation. AB - PURPOSE OF REVIEW: Androgen deprivation therapy remains the cornerstone of treatment for advanced prostate cancer. There are significant adverse effects associated with prolonged androgen deprivation therapy, with recent findings associating it with the metabolic syndrome and its associated health risks. RECENT FINDINGS: A review of the adverse pathologic effects of testosterone deprivation is performed. Recent findings associate androgen deprivation therapy with endocrine and metabolic disarray, specifically the metabolic syndrome, resulting in increased rates of diabetes and myocardial infarction in patients on testosterone deprivation. SUMMARY: The increased incidence of the metabolic syndrome in patients receiving androgen deprivation therapy and the associated morbidity and possibly mortality require significant investigation into alternatives to complete testosterone deprivation and ways to counteract these adverse effects. PMID- 17921779 TI - Therapeutic testis biopsy for sperm retrieval. AB - PURPOSE OF REVIEW: The introduction of IVF and intracytoplasmic sperm injection revolutionized the treatment of male infertility by requiring a minimal number of sperm to achieve pregnancy. We describe the various methods for testicular sperm retrieval in different types of azoospermia. RECENT FINDINGS: Different techniques for surgical sperm retrieval are available, including testicular sperm aspiration, single, multisite and microdissection testicular sperm extraction, as well as percutaneous and microsurgical epididymal sperm aspiration. While these methods have similar retrieval outcomes in obstructive azoospermia, testicular sperm extraction procedures appear to be more beneficial in cases of nonobstructive azoospermia. A reliable algorithm for predicting the outcome of microdissection testicular sperm extraction using clinical parameters is still lacking, and may be difficult to develop, given the fact that this method is designed to detect and sample even minute areas of focal spermatogenesis. SUMMARY: A variety of surgical sperm retrieval procedures can be performed for the purpose of subsequent or simultaneous IVF/intracytoplasmic sperm injection. The decision regarding the type of procedure should be based on the type of azoospermia, specific clinical circumstances, as well as on the surgeon's preference and experience. PMID- 17921780 TI - Bibliography. Current world literature. Reconstructive surgery. PMID- 17921783 TI - Body-mind learning: a lesson for the mind from muscle. PMID- 17921784 TI - The final frontier: oxygen flux into muscle at exercise onset. AB - In humans at exercise onset, intramuscular phosphocreatine decreases immediately, whereas muscle oxygen (O2) uptake seems to rise after a delay of up to 15 s which is inconsistent with models of metabolic control. Novel microcirculatory investigations reveal that elevated capillary-to-myocyte O2 flux in rat muscle is, in fact, initiated simultaneously with contractions. PMID- 17921786 TI - Electrical stimulation as a modality to improve performance of the neuromuscular system. AB - Transcutaneous neuromuscular electrical stimulation (NMES) can modify the order of motor unit recruitment and has a profound influence on the metabolic demand associated with producing a given muscle force. Because of these differences, interventions that combine NMES with voluntary contractions can provide beneficial outcomes for some individuals. The adaptations evoked by NMES are not confined to the activated muscle but also involve neural adaptations through reflex inputs to the spinal cord and supraspinal centers. PMID- 17921785 TI - Elastic properties of active muscle--on the rebound? AB - During active lengthening and shortening, muscles exhibit a variety of time dependent spring properties, including load-dependent and nonlinear stiffness. These properties can be explained as interactions between a spring element and cycling cross bridges within muscle sarcomeres. Several lines of evidence suggest a role for the giant protein titin in active muscle, but specific mechanisms remain to be elucidated. PMID- 17921787 TI - Engineered muscle: a tool for studying muscle physiology and function. AB - Recent advances in skeletal muscle tissue engineering have resulted in an in vitro tissue model that can be used for studying the effects of genetic alterations, pharmacological interventions, and exercise on muscle physiology and function. Here, we present applications for this technology to further our understanding of the molecular mechanisms underlying skeletal muscle adaptation in response to exercise. PMID- 17921788 TI - Exercise-induced alterations in muscle lipid metabolism improve insulin sensitivity. AB - Exercise is a key component for the successful management of many obesity-related metabolic complications, including insulin resistance. This review addresses the effect of chronic and acute endurance exercise on insulin action in obesity and the role of exercise-induced alterations in fatty acid partitioning within the muscle cell on insulin sensitivity. PMID- 17921789 TI - Admission of rheumatoid arthritis patients to rehabilitation hospitals or units after lower extremity arthroplasty. PMID- 17921790 TI - The impact of rheumatoid arthritis on rehabilitation outcomes after lower extremity arthroplasty. AB - BACKGROUND: Medical rehabilitation after lower extremity arthroplasty is an integral part of recovery and a critical step in returning to independent mobility. We hypothesized that rehabilitation may take longer for patients with rheumatoid arthritis (RA) versus osteoarthritis (OA) because joint pain, swelling, and deformities are generally worse among persons with RA. OBJECTIVES: To determine the impact of RA on length of rehabilitation stay and rehabilitation functional status gain after arthroplasty. METHODS: We conducted a retrospective cohort analysis using a national registry of US medical rehabilitation inpatients admitted after a lower extremity arthroplasty between 1994 and 2001. Sample included 1361 patients with RA and 26,096 patients with OA. The main outcome measure was functional status gain as assessed by the functional independence measure (FIM). Our primary analytic method was linear regression. Covariates were age, gender, race/ethnicity, other comorbidity, admission FIM, and site of arthroplasty. RESULTS: Mean length of stay for patients with RA was 11.3 +/- 7.1 days (mean +/- standard deviation) versus 10.3 +/- 6.5 days for those with OA. Mean weekly gain was 18.6 +/- 12.1 for patients with RA versus 20.6 +/- 12.0 for those with OA. After adjusting for covariates, RA was associated with longer stay (0.7 day) and lower FIM gain (2.6). CONCLUSIONS: RA was associated with longer length of rehabilitation stay and lower FIM gain in patients with lower extremity arthroplasty. Such patients may require additional monitoring to ensure sufficient rehabilitation. PMID- 17921791 TI - Efficacy and safety of topical nimesulide in the treatment of knee osteoarthritis. AB - BACKGROUND: Osteoarthritis (OA) affects mainly older people who are more sensitive to adverse effects of classic nonsteroidal anti-inflammatory drugs. Recent publications indicate that topically applied nonsteroidal anti inflammatory drugs are effective and much safer than their oral analogues. OBJECTIVES: In this double-blind, randomized, placebo-controlled study design, we aimed to investigate whether topical nimesulide treatment has any beneficial effect in knee OA patients. METHODS: Seventy-four adult knee OA outpatients were enrolled. We used Western Ontario and McMaster Universities OA Index (WOMAC), Nottingham Health Profile (NHP), and patient and physician global satisfaction scores. WOMAC and NHP scores were measured at the initial and final visit. Treatment group received topical nimesulide gel 1% on the knee skin 3 times a day whereas placebo group received an identical-appearing gel for 30 days. RESULTS: There was a significant improvement in the nimesulide treatment group for all 3 parameters and overall score of WOMAC between pretreatment and posttreatment values. The overall WOMAC scores was significantly better than placebo (P = 0.03), but physical functioning, stiffness, and pain scales did not reach statistical significance. For the NHP scores there was an improvement at "energy level," "pain," "physical motion," and "NHP distress" scores in the treatment group whereas no improvement was found in the placebo group. Between-group differences were not significant. Both patient and physician satisfaction scores were significantly better in the treatment group. CONCLUSION: This study shows that topical nimesulide gel can have beneficial effects and can improve quality of life in patients with knee OA. PMID- 17921792 TI - Partial C4 deficiency in juvenile idiopathic arthritis patients. AB - OBJECTIVES: C4 is encoded by 2 distinct but closely linked loci within the major histocompatibility complex locus on human chromosome 6. C4A deficiencies have been associated with autoimmune disease and C4B with increased frequency of infection. C4 deficiencies have rarely been associated with juvenile idiopathic arthritis (JIA). Our aim was to investigate the prevalence of deficiencies in C4 allotypes in JIA patients. METHODS: We evaluated 61 patients [35 JIA patients, 15 systemic lupus erythematosus patients, 9 rheumatoid arthritis patients, and 2 mixed connective tissue disease (CTD) patients] for C4 deficiency. Genomic DNA was isolated from whole blood and subjected to polymerase chain reaction using sequence-specific primers for C4 allotypes. RESULTS: We found 5 JIA patients with C4 deficiencies. Two IgM rheumatoid factor-positive JIA polyarthritis patients had C4 deficiencies, one with complete C4A deficiency and another with partial C4A and complete C4B deficiency. Two oligoarthritis patients displayed partial C4B deficiencies, and complete C4B deficiency was revealed in 1 IgM rheumatoid factor-negative polyarthritis patient. Three patients had histories of recurrent infections and 2 demonstrated a more severe disease course. Disease controls showed 8 systemic lupus erythematosus patients had partial C4 deficiencies, whereas no deficiencies were revealed in the rheumatoid arthritis or mixed CTD patients. CONCLUSIONS: Defects in the complement system have been implicated in the pathogenesis of CTD. However, the specific role of C4 in JIA is not clear. We demonstrate partial C4 deficiencies in 5 JIA patients. Our findings suggest an association between C4 deficiency and another CTD, JIA, as well as with disease severity and recurrent infections. PMID- 17921793 TI - Systemic lupus erythematosus in a multiethnic US cohort (LUMINA L II): relationship between vascular events and the use of hormone replacement therapy in postmenopausal women. AB - OBJECTIVES: To examine the influence of hormone replacement therapy (HRT) in the occurrence of vascular arterial and venous thrombotic events in postmenopausal women with systemic lupus erythematosus (SLE). PATIENTS AND METHODS: SLE women aged > or =16 years, disease duration < or =5 years from LUMINA, a multiethnic, longitudinal outcome study, were included. Menopause was defined at disease onset as the presence of amenorrhea >6 months and/or oophorectomy, and/or increased follicle stimulating hormone values, and/or HRT use regardless of the presence or absence of climacteric symptoms (hot flashes). Patients were divided into HRT ever users and nonusers. Patients with positive antiphospholipid antibodies (n = 9) or vascular arterial events (n = 1) occurring before HRT use were excluded. The occurrence of vascular arterial and venous thrombotic events was compared between HRT users and HRT nonusers and its role examined by logistic regression after adjusting for "confounding by indication" using propensity score or logistic regression analyses. RESULTS: Seventy-two postmenopausal women, 32 (44%) HRT users and 40 (56%) HRT nonusers, were studied. HRT use was associated with fewer vascular arterial but not venous thrombotic events (P = 0.021) in the univariable analyses. However, after adjusting for the propensity score, HRT use was no longer significant (P = 0.064). Comparable results were obtained by logistic regression. CONCLUSIONS: HRT use was not associated with the occurrence of vascular arterial events in the LUMINA patients. HRT use in women with SLE should be individualized, but our data suggest its use may be safe if antiphospholipid antibodies are not present or vascular arterial events have not previously occurred. PMID- 17921794 TI - Rapid onset of muscle weakness (rhabdomyolysis) associated with the combined use of simvastatin and colchicine. AB - We report a case of a patient with mild chronic renal insufficiency who had been taking simvastatin for over a year and developed acute weakness within 3 weeks after the start of treatment with colchicine for acute gouty bursitis. Profound muscle weakness of lower extremities with inability to stand up and/or walk was present. Elevated muscle enzymes and findings on electromyography were consistent with myopathy. Rapid improvement in muscle strength accompanied by prompt resolution of abnormal elevation of muscle enzymes followed cessation of both medications. Both colchicine and statin therapy may be associated with myopathy, which usually occurs after several months of therapy. The concomitant use, however, of colchicine and statin has been associated with the rapid onset of muscle weakness. Four patients with similar clinical and laboratory characteristics to our patient's after the combined use of colchicine and statins have been described in the literature. Patients receiving combination therapy with colchicine and simvastatin, particularly in the presence of renal insufficiency, should be monitored for the development of myopathy, including rhabdomyolysis. PMID- 17921795 TI - Leontiasis ossea: a paleopathologic case report. AB - We describe the archeological and imaging findings of a unique specimen (skull and mandible) with leontiasis ossea (LO) that is on display in the National Museum of Anthropology and History in Mexico City. The specimen shows diffuse and irregular periosteal bone proliferation, which produces a grossly nodular appearance involving the neurocranium and the facial skeleton. Plain radiography and helical computed tomography revealed generalized hyperostosis obliterating the maxillary and sphenoidal sinuses and 2 exuberant bony masses arising from the maxilla with encroachment of the anterior nasal aperture.Currently, LO is a purely descriptive term applied to a variety of osseous conditions that have in common hyperostosis of craniofacial bones leading to a leonine appearance. Clinicians who see such lionlike facies should consider the main causes of LO, which include renal osteodystrophy, Paget disease and, as most likely in this specimen, fibrous dysplasia. PMID- 17921796 TI - Hypothyroidism presenting as puzzling myalgias and cramps in 3 patients. AB - Cramps and myalgias have many etiologies. Three cases of hypothyroidism presenting with musculoskeletal symptoms are described. Each case was different in presentation, but all had no weakness but high creatine kinase levels. Treatment with thyroxine resolved all symptoms. These patients emphasize the importance of considering hypothyroidism in anyone with unexplained muscle symptoms. PMID- 17921797 TI - Severe pulmonary hypertension in pediatric primary Sjogren syndrome: a case report. AB - A 9-year-old girl with a history of xerostomia and recurrent bilateral parotid gland enlargement presented with purpura of the lower limbs and exertional dyspnea. She had hyperglobulinemia, positive ANA, SSA, and SSB and an abnormal Shirmer test leading to a diagnosis of primary Sjogren syndrome. Cardiologic and imaging investigations revealed severe isolated pulmonary hypertension and ruled out pulmonary fibrosis. Prednisolone and cyclophosphamide, together with anticoagulant and vasodilatory drugs therapy, induced a significant improvement of exertional dyspnea and lowered pulmonary artery pressure. This case and reports from the literature suggest that immune mechanisms, not just vasospasm, can be factors in some pulmonary hypertension. PMID- 17921798 TI - Laryngeal involvement in systemic lupus erythematosus: report of two cases. AB - Systemic lupus erythematosus is a multisystem autoimmune disorder. Joint and skin involvements are the most frequent presenting features. Laryngeal involvement, however, is extremely rare. Symptoms of laryngeal involvement may range from mild hoarseness to life-threatening respiratory distress. In this article, 2 patients with systemic lupus erythematosus and laryngeal involvement are presented. The first patient had hoarseness and noisy respiration, and the second had cough and respiratory distress. Otorhinolaryngological examination revealed laryngeal inflammation in both. Our cases responded well to systemic corticosteroids. Patients with symptoms such as hoarseness, foreign body sensation in throat, and respiratory distress should be evaluated by an otorhinolaryngologist to rule out involvement of the laryngeal structures. PMID- 17921799 TI - Newer imaging technology for the diagnosis of early RA and monitoring of inflammation and joint damage. PMID- 17921801 TI - Basic science for the clinician 46. Molecular biologic agents in development: possible clinical consequences to rheumatologists. AB - The molecular biologic revolution has been upon us for only a few decades, and yet diagnostics and therapeutics have been, well, revolutionized. No aspect of the practice of rheumatology has been left untouched and our field is virtually unrecognizable compared with only a decade ago. No list of molecular biologic can be complete, as there are agents in development that have not yet seen clinical trials, the length and width of the "pipelines" of the various companies and academic allies being what they are. What follows is a brief review of some of the approaches that can and are being taken and some of the molecules already making some noise. Small molecules will not be reviewed, the emphasis here being on antibodies and larger molecules, cytokines among them. PMID- 17921802 TI - Osteopoikilosis with abnormal bone scan: long-term follow-up. PMID- 17921803 TI - Tophaceous gout and chronic kidney disease. PMID- 17921804 TI - Resolution of treatment-related arthralgias and serologic findings with a switch of TNF antagonist therapies in a patient with psoriatic arthritis. PMID- 17921805 TI - Are anti-TNFalpha therapies too good? An audit of patients' compliance. PMID- 17921806 TI - Lamivudine in the treatment of hepatitis B virus-related polyarteritis nodosa. PMID- 17921807 TI - Generalized nonspecific pustular lesions in Tietze's syndrome. PMID- 17921808 TI - Telomere dysfunction in hypertension. AB - Aging is a major risk factor for hypertension and associated cardiovascular disease. In most proliferative tissues, aging is characterized by shortening of the DNA component of telomeres, the specialized genetic segments that cap the end of eukaryotic chromosomes and protect them from end-to-end fusions. By inducing genomic instability, replicative senescence and apoptosis, telomere shortening is thought to contribute to organismal aging and to the development of age-related diseases. Here, we review animal and human studies that have investigated the possible links between telomere ablation and the pathogenesis of hypertension and related target organ damage. Although evidence is mounting that alterations in telomerase activity and telomere shortening may play a role in the pathogenesis of hypertension, additional studies are required to understand the molecular mechanisms by which telomere dysfunction and hypertension are functionally connected. As our knowledge on this emerging field grows, the challenge will be to ascertain whether all this information might translate into clinical applications. PMID- 17921810 TI - Malignant hypertension and the angiotensinogen gene. PMID- 17921809 TI - Incidence of cardiovascular events in white-coat, masked and sustained hypertension versus true normotension: a meta-analysis. AB - OBJECTIVE: To perform a meta-analysis on the incidence of cardiovascular events in white-coat hypertension (WCHT), masked and sustained hypertension in comparison with true normotension. METHODS: We searched for individual studies, in which the adjusted relative risk of incident cardiovascular events was assessed in patients with WCHT, masked and sustained hypertension versus normotension in the same study population. For each type of hypertension, the weighted overall hazard ratio (HR) and 95% confidence intervals (CI) were calculated. RESULTS: Seven studies were identified, involving a total of 11 502 participants. Four studies were performed in the population, two in primary care and one in specialist care. Two studies were exclusively on treated hypertensive patients; individuals on antihypertensive treatment were included in all the other studies except one. Cut-off blood pressure was 140/90 mmHg for office blood pressure in all studies and 135/85 mmHg (83 in one study) for out-of-office blood pressure. The average age of the study populations was 63 years; 53% were men. The endpoint consisted of cardiovascular death in one study and of various aggregates of fatal and non-fatal cardiovascular events in the others. During an average follow-up of 8.0 years, there were 912 first cardiovascular events. The overall adjusted HR versus normotension was 1.12 (95% CI 0.84-1.50) for WCHT (P = 0.59), 2.00 (1.58-2.52) for masked hypertension (P < 0.001), and 2.28 (1.87-2.78) for sustained hypertension (P < 0.001). CONCLUSION: The meta-analysis indicates that the incidence of cardiovascular events is not significantly different between WCHT and true normotension, whereas the outcome is worse in patients with masked or sustained hypertension. PMID- 17921811 TI - Endothelial vasodilatory cannabinoid receptor in the human pulmonary artery: a future option in the therapy of pulmonary hypertension? PMID- 17921812 TI - Pseudopheochromocytoma. PMID- 17921814 TI - Prevalence of hypertension in schoolchildren based on repeated measurements and association with overweight. AB - OBJECTIVE: Most studies assess the prevalence of hypertension in pediatric populations based on blood pressure (BP) readings taken on a single visit. We determined the prevalence of hypertension measured on up to three visits in a Swiss pediatric population and examined the association between hypertension and overweight and selected other factors. METHODS: Anthropometric data and BP were measured in all children of the sixth school grade of the Vaud canton (Switzerland) in 2005-2006. 'Elevated BP' was defined according to sex-specific, age-specific and height-specific US reference data. BP was measured on up to two additional visits in children with elevated BP. 'Hypertension' was defined as 'elevated BP' on all three visits. RESULTS: Out of 6873 children, 5207 (76%) participated [2621 boys, 2586 girls; mean (SD) age, 12.3 (0.5) years]. The prevalence of elevated BP was 11.4, 3.8 and 2.2% on first, second and thirds visits, respectively; hence 2.2% had hypertension. Among hypertensive children, 81% had isolated systolic hypertension. Hypertension was associated with excess body weight, elevated heart rate and parents' history of hypertension. Of the children, 16.1% of boys and 12.4% of girls were overweight or obese (CDC criteria, body mass index >or= 85th percentile). Thirty-seven percent of cases of hypertension could be attributed to overweight or obesity. CONCLUSIONS: The proportion of children with elevated BP based on one visit was five times higher than based on three measurements taken at few-week intervals. Our data re emphasize the need for prevention and control of overweight in children to curb the global hypertension burden. PMID- 17921813 TI - Is aldosterone a pro-arrhythmic hormone? PMID- 17921815 TI - Prevalence and determinants of hypertension in the Algerian Sahara. AB - BACKGROUND: In-Salah is a city-oasis located in the middle of the Algerian Sahara, a desert area whose drinking water has a high sodium content. No cardiovascular epidemiological studies have ever been conducted in this region. METHODS: A randomized sample of 635 men and 711 women, aged 40-99 years, was studied. Blood pressure measurements, combined with a clinical questionnaire that included educational and socio-economic data, and standard blood samples for the detection of dyslipidemia and diabetes mellitus, were collected. RESULTS: The mean age was 55 +/- 12 years. The prevalence of hypertension was 44% and was highly influenced by age, sex, skin colour, educational status, obesity and metabolic parameters. The higher prevalence of hypertension among black individuals was independent of socio-economic and educational levels, and of metabolic parameters. The presence of antihypertensive treatment was three times more frequent in women than in men, and there was no difference according to skin colour. Among treated subjects, 25% were well controlled, and this percentage was similar among both black and white individuals. CONCLUSION: Epidemiological studies in such an emergent population indicate that hypertension is a major public health problem. The high sodium content in drinking water in this region could play a major role in the development of hypertension. PMID- 17921816 TI - The M235T polymorphism in the angiotensinogen gene is associated with the risk of malignant hypertension in white patients. AB - BACKGROUND: Malignant hypertension can be considered an extreme phenotype of renin-mediated hypertension. Therefore, we compared the allelic frequencies of the angiotensinogen (AGT) M235T, angiotensin-converting enzyme insertion/deletion (ACE I/D) and angiotensin II-type I receptor (AT1R) A1166C polymorphisms in malignant hypertensive patients with hypertensive and normotensive controls. METHODS: A total of 101 consecutive patients between 1995 and 2005 admitted to a large university hospital fulfilled the criteria for malignant hypertension. Seventy-five patients (74%) were compared with 150 hypertensive and 150 normotensive controls, randomly selected from a population study and individually matched on age, sex and ethnicity. RESULTS: The odds of malignant hypertension in white subjects with the TT genotype of the AGT M235T polymorphism was 14.3 (5.5 37) compared to hypertensive controls, and 9.4 (3.8-23.2) compared to normotensive controls. Adjustment for age, sex, smoking and antihypertensive therapy did not affect this association. The association of AGT M235T with malignant hypertension was not significant in blacks. In patients with malignant hypertension, the TT genotype was associated with more severe renal dysfunction and microangiopathic haemolysis. No differences were found in allele frequencies of the ACE I/D or the AT1R A1166C polymorphisms between study groups. CONCLUSIONS: The TT genotype of AGT M235T is associated with malignant hypertension in whites, carriers having an odds of approximately 10 to 1 compared to hypertensive and normotensive controls. These observations may provide a better understanding of the pathophysiology of malignant hypertension and offer possibilities for identifying patients at risk. Larger association or linkage studies are needed for a more detailed risk assessment. PMID- 17921817 TI - Gly460Trp alpha-adducin gene polymorphism and endothelial function in untreated hypertensive patients. AB - OBJECTIVES: Endothelium-dependent vasodilatation is impaired in essential hypertension. Besides traditional and emerging cardiovascular risk factors, genetic factors may also promote deleterious alterations of endothelial physiology. The aim of the present study was to investigate the relationship between the 460Trp allele of ADD1 and endothelium-dependent vasodilation in 110 never-treated hypertensive patients. METHODS: Forearm blood flow (FBF) was measured during intra-arterial infusion of acetylcholine (ACh) and sodium nitroprusside (SNP) at increasing doses. Analysis of endothelium-dependent and endothelium-independent vasodilation was tested according to ADD1 genotype. RESULTS: The FBF values at the three incremental doses of ACh were 5.22 +/- 0.24 (+76%), 8.64 +/- 0.45 (+193%) and 14.74 +/- 0.71 (+395%) ml/100 ml of tissue per min for Gly460Gly and 4.63 +/- 0.20 (+51%), 6.84 +/- 0.36 (+123%) and 11.22 +/- 3.8 (+269%) ml/100 ml of tissue per min for 460Trp. Thus, ACh-stimulated FBF was significantly reduced in hypertensive subjects carrying the 460Trp allele of ADD1 (P < 0.001). SNP-stimulated FBF was not affected by ADD1. CONCLUSIONS: The main finding in this study was that in essential hypertensives the 460Trp allele of ADD1 is strongly associated with an impaired endothelium-dependent vasodilation, a powerful predictor of cardiovascular risk. PMID- 17921818 TI - Identification of the vasodilatory endothelial cannabinoid receptor in the human pulmonary artery. AB - BACKGROUND: The endocannabinoid anandamide is implicated in the pathogenesis of hypotension in haemorrhagic, endotoxic, and cardiogenic shock. It has been demonstrated in animal, but not in human, vessels that the vasodilatory effects of anandamide and abnormal cannabidiol are partially mediated by an as yet unidentified endothelial cannabinoid receptor. Our study was performed to examine the influence of abnormal cannabidiol on the human pulmonary artery. METHODS: Isolated human pulmonary arteries were obtained from patients without clinical evidence of pulmonary hypertension during resection of lung carcinoma. Vasodilatory effects of abnormal cannabidiol were examined on endothelium-intact vessels preconstricted with serotonin or potassium chloride. RESULTS: Anandamide and abnormal cannabidiol relaxed serotonin-preconstricted vessels concentration dependently. The effect of abnormal cannabidiol was reduced by endothelium denudation, pertussis toxin and two antagonists of the novel endothelial receptor, cannabidiol and O-1918, but not by the nitric oxide synthase inhibitor L-NAME given together with the cyclooxygenase inhibitor indomethacin. It was also diminished by blockade of calcium-activated potassium channels by the nonselective blocker tetraethylammonium or by combination of selective blockers of small (apamin) and intermediate and large (charybdotoxin) conductance calcium activated potassium channels. The potency of abnormal cannabidiol to relax vessels was lower in potassium chloride than in serotonin-preconstriced preparations. CONCLUSIONS: Abnormal cannabidiol relaxes human pulmonary arteries in an endothelium-independent and endothelium-dependent manner. The latter component is probably mediated via the putative endothelial cannabinoid receptor, activation of which may release endothelium-derived hyperpolarizing factor, which in turn acts via calcium-activated potassium channels. Abnormal cannabidiol is behaviourally inactive; it may have a therapeutic implication in vascular diseases, especially in the treatment of pulmonary hypertension. PMID- 17921819 TI - The polymorphonuclear leukocyte contributes to the development of hypertension in the Sabra rat. AB - BACKGROUND: We previously showed that priming of the polymorphonuclear leukocyte (PMNL), inflammation and oxidative stress antecede the development of hypertension in the Sabra rat model of hypertension. The actual role of PMNLs and PMNL-mediated oxidative stress and inflammation in the development of hypertension in this model has remained, however, unresolved. OBJECTIVE: The aim of our study was to test the hypothesis that PMNLs and that the PMNL-associated NADPH oxidase contribute to the development of hypertension in the Sabra rat model. METHODS: To determine the contribution of the PMNL to the development of hypertension, we depleted Sabra hypertension-prone (SBH/y) animals from PMNLs with an anti-PMNL antibody, salt-loaded them and monitored their blood pressure over a period of 30 days. To determine the contribution of the NADPH oxidase on the development of hypertension, we inhibited the activity of this enzyme with phenylarsine oxide or apocynin in SBH/y rats while salt-loading the animals and followed the course of their blood pressure over 60 days. RESULTS: PMNL depletion attenuated significantly the development of hypertension in SBH/y rats. Inhibition of NADPH oxidase with phenylarsine oxide and apocynin markedly inhibited the development of hypertension in SBH/y rats, as well as decreased the rate of superoxide release, the level of PMNL CD11b and the PMNL count. CONCLUSION: These data are consistent with a significant contribution of PMNLs to the development of hypertension, and suggest that the mechanism may be related, at least in part, to PMNL-mediated oxidative stress and inflammation. PMID- 17921820 TI - Superoxide and its interaction with nitric oxide modulates renal function in prehypertensive Ren-2 transgenic rats. AB - OBJECTIVE: The present study was performed to examine the role of superoxide (O2*) and its interaction with nitric oxide (NO) in the regulation of renal function in prehypertensive heterozygous Ren-2 transgenic rats (TGR). METHODS: Renal responses to the O2* scavenger, tempol (150 microg/min per 100 g), and/or the NO synthase inhibitor, nitro-L-arginine methylester (L-NAME; 5 microg/min per 100 g), infused alone or in combination directly into the renal artery were evaluated in anesthetized heterozygous male TGR and aged-matched Hanover Sprague Dawley rats (HanSD). RESULTS: There were no differences in arterial pressure (122 +/- 3 versus 115 +/- 2 mmHg), renal plasma flow (RPF; 2.09 +/- 0.1 versus 2.07 +/ 0.1 ml/min per g), glomerular filtration rate (GFR; 0.73 +/- 0.1 versus 0.74 +/- 0.1 ml/min per g) or sodium excretion (0.63 +/- 0.13 versus 0.67 +/- 0.16 micromol/min per g) between TGR and HanSD. Tempol alone caused significant increases in RPF and GFR (10 +/- 4% and 12 +/- 2%, respectively) in TGR but not in HanSD. Tempol also caused greater sodium excretory responses in TGR compared to HanSD (112 +/- 16% versus 43 +/- 7%; P < 0.05). 8-Isoprostane excretion was significantly higher in TGR than in HanSD (10.2 +/- 0.8 versus 6.5 +/- 0.7 pg/min per g), which was attenuated by tempol. L-NAME caused greater decreases in RPF and GFR in TGR (-34 +/- 4% and -22 +/- 4%, respectively) than in HanSD (-19 +/- 3% and -10 +/- 4%, respectively). Co-infusion of tempol partially attenuated the renal hemodynamic and excretory responses to L-NAME in TGR. CONCLUSIONS: These data suggest that the enhanced O2* activity and its interaction with NO during the prehypertensive phase in TGR modulates renal hemodynamic and excretory function, which may contribute to the development of hypertension in this transgenic rat model. PMID- 17921821 TI - A generalized arterial transfer function derived at rest underestimates augmentation of central pressure after exercise. AB - OBJECTIVES: Peripheral exercise blood pressure and resting central blood pressure are considered more relevant to cardiovascular health than resting peripheral blood pressure. Central exercise blood pressure may well be an even more useful measure, but there is no simple non-invasive means of determining it. The aim of the present study was to establish whether the estimation of central blood pressure from peripheral blood pressure using a transfer function derived at rest, would hold after aerobic exercise. METHODS: Thirty healthy young men were studied before and immediately (< 1 min) and 10 min after 15 min bicycle exercise at 65-70% of maximum heart rate. Simultaneous carotid and radial artery waveforms were recorded, and radial-to-carotid generalized transfer functions (GTF) were calculated using Fourier analysis for rest and immediately postexercise. Central systolic blood pressure (SBP) and augmentation index (AIx) were calculated for measured and derived waves. RESULTS: The resting GTF underestimated central SBP and AIx immediately (-5.8 +/- 2.1 mmHg, P = 0.01; -8.3 +/- 2.9%, P = 0.008) and 10 min after (-2.0 +/- 0.7 mmHg, P = 0.008; -7.0 +/- 2.1%, P = 0.003) exercise. No significant bias was found between measured and derived (using resting GTF) carotid values at rest. The use of an exercise-specific GTF resulted in no specific bias immediately or 10 min after exercise, although it overestimated blood pressure and AIx at rest (2.5 +/- 1.0 mmHg, P = 0.02; 11.3 +/- 3.0%, P = 0.001). CONCLUSION: A peripheral-to-central arterial GTF derived at rest significantly underestimates key measures of central arterial pressure immediately after exercise, and pressure estimations may be improved by the use of an exercise-specific GTF. PMID- 17921822 TI - Role of heart failure etiology on arterial wave reflection in heart transplant recipients: relation with C-reactive protein. AB - OBJECTIVES: Aortic augmentation index (AIa), a measure of arterial pressure wave reflection related to central and peripheral arterial stiffness, is elevated in many heart transplant recipients. We investigated whether the increase in wave reflection observed in some heart transplant recipients is influenced by the etiology of antecedent heart failure and circulating pro-inflammatory proteins early in the post-transplantation period. METHODS: Two months after heart transplantation, 20 heart transplant recipients underwent noninvasive measurement of aortic pressure and wave reflection properties and measurement of plasma pro inflammatory proteins. RESULTS: AIa adjusted to a heart rate of 75 beats/min (AIaHR75) was higher in heart transplant recipients with ischemic (n = 12) compared with nonischemic (n = 8) heart failure (P < 0.01). Similarly, circulating C-reactive protein, a marker of systemic inflammation and an independent predictor of allograft vasculopathy and death in heart transplant recipients, was higher in heart transplant recipients with ischemic than with nonischemic heart failure (log-transformed, P < 0.05). Moreover, there was a significant relation between log C-reactive protein and AIaHR75 (r = 0.68, P < 0.05), augmented pressure (r = 0.60, P < 0.01), roundtrip time of the reflected wave to the peripheral reflecting sites and back (r = -0.62; P < 0.01), and left ventricular wasted energy (r = 0.55, P < 0.01). Multiple regression analysis revealed that log C-reactive protein explained 43% of the variance in AIaHR75 and the difference in AIaHR75 between groups was abolished when adjusted for log C reactive protein. CONCLUSIONS: Heart transplant recipients with antecedent ischemic heart failure demonstrated increased AIaHR75 compared with nonischemic heart transplant recipients and AIaHR75 was associated with higher circulating C reactive protein concentration. Whether elevated arterial wave reflection and associated systemic low-grade inflammation early after transplantation have clinical implications in ischemic heart transplant recipients requires further investigation. PMID- 17921823 TI - Sympathetic activity, assessed by power spectral analysis of heart rate variability, in white-coat, masked and sustained hypertension versus true normotension. AB - OBJECTIVE: To assess, in a population-based approach, sympathetic nervous system activity by the use of power spectral analysis of heart rate variability, in normotension, white-coat hypertension, masked hypertension and sustained hypertension. METHODS: The electrocardiographic RR interval was registered in the supine and standing positions and the low-frequency and high-frequency components of its variability were quantified. Cut-off values of 140/90 mmHg for conventional blood pressure and 135/85 mmHg for daytime ambulatory blood pressure were used to define the four blood pressure groups. RESULTS: After exclusion of patients with diabetes, myocardial infarction or treated hypertension, 1485 subjects with complete data remained for the analysis in the supine position. Age averaged 39 +/- 14 years; 54% were women. Conventional and ambulatory blood pressure averaged, respectively, 122 +/- 16/79 +/- 11 mmHg and 124 +/- 12/77 +/- 8 mmHg. After adjusting for demographic, anthropometric and lifestyle characteristics, the low-frequency to high-frequency ratio (geometric mean) averaged 0.81 in normotension and was significantly higher in white-coat hypertension (1.11; P < 0.001), based on a higher low-frequency component and a lower high-frequency component (P < 0.01). This ratio was not significantly different between normotension, masked hypertension (0.97) and sustained hypertension (0.93). The adjusted standing-to-supine ratio of the high-frequency component (geometric mean) was significantly higher in sustained hypertension (0.50) than in normotension (0.39; P < 0.01), but not in white-coat (0.40) and masked hypertension (0.45). CONCLUSION: The findings at rest are compatible with increased sympathetic activity and decreased parasympathetic modulation in white coat hypertension, with normal autonomic cardiac regulation in masked and sustained hypertension. In addition, sustained hypertension is characterized by a blunted decrease of the high-frequency component on standing. PMID- 17921824 TI - Sympathoadrenal function in patients with paroxysmal hypertension: pseudopheochromocytoma. AB - OBJECTIVES: The causes of paroxysmal hypertension in patients in whom pheochromocytoma has been excluded ('pseudopheochromocytoma') usually remain unclear. Blood pressure disturbances and symptoms of catecholamine excess in these patients may reflect activation of the sympathetic nervous and adrenal medullary systems. We therefore examined sympathoadrenal function in patients with pseudopheochromocytoma compared with age-matched control subjects in whom there was no suspicion of pheochromocytoma. METHODS: Plasma catecholamines and hemodynamics were examined in response to intravenous glucagon, yohimbine, and trimethaphan in 11 patients with pseudopheochromocytoma and a comparison group of nine normotensive and five hypertensive volunteers. Adrenomedullary function was also assessed by abdominal F-fluorodopamine positron emission tomography and measurements of plasma metanephrine, the O-methylated metabolite of epinephrine. RESULTS: Compared with controls, patients with pseudopheochromocytoma had normal plasma concentrations of norepinephrine, but 120% higher (P < 0.05) baseline plasma concentrations of epinephrine, 80% higher (P < 0.01) baseline plasma concentrations of metanephrine, and sixfold larger (P < 0.05) increases in plasma epinephrine after glucagon. Adrenal 18F-fluorodopamine-derived radioactivity did not differ between groups. Compared with changes in plasma norepinephrine, falls in blood pressure after trimethaphan were 13-fold larger (P < 0.005) and increases in blood pressure after yohimbine were threefold larger (P < 0.01) in pseudopheochromocytoma patients than in controls. CONCLUSION: Patients with pseudopheochromocytoma exhibit a pattern of normal sympathetic noradrenergic outflow, adrenomedullary activation, and augmented blood pressure responses to changes in the sympathoneural release of norepinephrine. PMID- 17921825 TI - Association of suboptimal blood pressure control with body size and metabolic abnormalities. AB - BACKGROUND: Blood pressure control is disappointingly suboptimal in populations. Whether metabolic abnormalities influence blood pressure control is unclear. We evaluated the relationship between metabolic risk factors and blood pressure control in a large population of patients with hypertension. METHODS: From our Hypertension Centre, 4551 subjects (43.4% women; age 51 +/- 12 years) were selected with available data for metabolic and cardiovascular evaluation (no prevalent cardiovascular disease), at the last control visit. A modified Adult Treatment Panel III definition of metabolic syndrome was adopted changing waist girth for body mass index (>or= 30 kg/m2). Blood pressure was considered controlled when supine office blood pressure was below 140/90 mmHg, or uncontrolled if this target was not achieved. Blood pressure control has been evaluated in relation to metabolic risk factors, adjusting for age, sex, and the number of antihypertensive medications. RESULTS: The metabolic syndrome phenotype was found in 1444 individuals (31.72%). The probability of uncontrolled blood pressure was 43% higher in patients with the metabolic syndrome than in those without, independently of covariates. This probability was also confirmed in 728 untreated patients. The probability of uncontrolled blood pressure significantly and independently increased with the increasing number of metabolic risk factors. Uncontrolled blood pressure was also independently associated with the prescription of more medications. CONCLUSION: Insufficient control of blood pressure is independently associated with the presence of the metabolic syndrome. Blood pressure control worsens with the increasing number of metabolic risk factors associated with hypertension, despite the use of a greater number of medications. PMID- 17921826 TI - Decreased flow-mediated dilation is present 1 year after a pre-eclamptic pregnancy. AB - OBJECTIVE: Pre-eclamptic toxaemia is associated with inflammation and vascular endothelial dysfunction. As women who have had pre-eclamptic toxaemia are at an increased risk of cardiovascular disease, we hypothesized that these abnormalities are persistent. METHODS: Eighteen women with a history of pre eclamptic toxaemia and 17 age-matched controls were enrolled. All underwent non invasive ultrasound examination of the brachial artery for an evaluation of flow mediated vasodilatation (FMD). The ambulatory blood pressure measurement (ABPM), and plasma concentrations of lipoproteins, inflammation markers, adhesion molecules, glucometabolic and haemostatic factors were determined. RESULTS: Women with a history of pre-eclamptic toxaemia had lower FMD compared with controls (2.5 +/- 2.9 versus 10.3 +/- 2.0%, P < 0.0001). ABPM showed higher systolic, diastolic and mean arterial blood pressures during daytime in the pre-eclamptic toxaemia group than in controls (123 +/- 9, 81 +/- 6 and 95 +/- 6 mmHg versus 116 +/- 9, 76 +/- 7 and 90 +/- 7 mmHg, respectively, all P < 0.05). Among the biochemical determinations, a high value of the homeostasis model assessment of insulin resistance was calculated at 1.3 (1.1-2.1) median [interquartile range (IQR)] in the pre-eclamptic toxaemia group and 1.0 (0.7-1.3) in controls (P < 0.01), and when adjusted for body mass index there was still a significant difference between groups (P < 0.05). No significant differences were found for other metabolic and haemostatic factors. CONCLUSION: Women with a previous history of pre-eclamptic toxaemia have decreased FMD compared with women with a previous normal pregnancy. This perturbation is a proof of an abnormal state still present 1 year after delivery. PMID- 17921827 TI - Induction of C1q expression in glomerular endothelium in a rat model with arterial hypertension and albuminuria. AB - OBJECTIVE: Increased urinary albumin excretion (UAE) represents an independent cardiovascular risk factor in the general population and particularly in patients with diabetes or arterial hypertension. It has been suggested that increased UAE may be related to a generalized endothelial dysfunction. We set out to identify candidate genes for increased UAE by glomerular transcriptome analysis in the Munich Wistar Fromter (MWF) genetic rat model with spontaneous hypertension and albuminuria. METHODS: First, we performed microarray expression analysis in isolated glomerular tissue in MWF with established albuminuria and normal Wistar rats. Second, in validation experiments and follow-up studies we focused on the identified upregulation of glomerular complement component C1q expression in MWF. RESULTS: Overall, 38 genes with a regulation score > 2 were differentially expressed in glomerular RNA. Quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR), in-situ hybridization and immunohistochemistry analysis revealed that C1q is indeed significantly upregulated in the glomerulus of MWF. Additionally, CD24, although not detected by the microarray experiment, was found to be differentially expressed in MWF glomeruli using quantitative real-time RT-PCR and immunohistochemstry. Interestingly, we could show for the first time that the glomerular endothelium represents the site of increased C1q and CD24 expression in MWF. In contrast, endothelial expression of this gene is low or absent in normotensive Wistar and in spontaneously hypertensive rats (SHR) without albuminuria. CONCLUSIONS: The induction of C1q and CD24 expression confined to the glomerular endothelium might represent a possible repair mechanism of the capillary wall damage. PMID- 17921828 TI - Association of cardiovascular risk factors with microalbuminuria in hypertensive individuals: the i-SEARCH global study. AB - OBJECTIVES: To define the prevalence of microalbuminuria (MAU) in hypertensive outpatients attending a cardiologist or internist (i-SEARCH A) and to compare hypertensive outpatients with or without coronary artery disease (CAD; i-SEARCH B). A secondary objective was to establish a correlation between MAU and known cardiovascular risk factors. METHODS: i-SEARCH was an international, observational study. which enrolled consecutive outpatients with hypertension. Patients with reasons for a false-positive MAU test were excluded. Main outcome measures were the prevalence of MAU as assessed using a dipstick test, hypertension co-morbidities, co-medication and presence of known cardiovascular risk factors. RESULTS: A total of 21,050 patients, from 26 countries, were included in the primary analysis. Overall prevalence of MAU was 58.4% (men > women), although there was considerable variation in prevalence across countries and continents (maximum 71% in Vietnam/Indonesia; minimum 53% in Germany/Switzerland). In multivariate analyses, predictors of MAU were identified to be male gender, high waist circumference, systolic blood pressure >or= 120 mmHg, diastolic blood pressure >or= 100 mmHg, creatinine clearance >or= 50 ml/min, and the presence of diabetes, congestive heart failure, CAD, history of cerebral pathology, peripheral arterial disease, dyspnoea or palpitations. MAU was present more often in patients with CAD than in patients without. CONCLUSIONS: MAU is extremely common in hypertensive outpatients worldwide, especially in patients with known cardiovascular risk factors. Given its importance as a strong, early and independent marker of increased cardiovascular risk in hypertension, the results of i-SEARCH mandate more rigorous MAU screening of hypertensive patients in clinical practice. PMID- 17921829 TI - Effects of age on hypertensive status in patients with chronic kidney disease. AB - OBJECTIVE: To evaluate effect of age on hypertensive status in chronic kidney disease (CKD). METHODS: We studied 459 prevalent CKD patients (stages 2-5, no dialysis), grouped by age (< 55, 55-64, 65-74, >or= 75 years), undergoing clinical blood pressure (CBP) and ambulatory blood pressure (ABP) measurement. RESULTS: Prevalence of diabetes, left ventricular hypertrophy and previous cardiovascular disease progressively increased with aging; glomerular filtration rate (GFR) and hemoglobin decreased. Achievement of CBP target decreased from 16% in patients < 55 years to 6% in those >or= 75 years (P = 0.023). ABP 24-h systolic rose while diastolic decreased, with a consequent pulse pressure increase from 45 +/- 8 to 65 +/- 14 mmHg (P < 0.0001). Age, proteinuria, diabetes, cardiovascular disease and anemia but not GFR predicted higher 24-h pulse pressure. CBP overestimated systolic/diastolic daytime ABP by 14 +/- 18/7 +/- 11 mmHg on average, a greater difference in older than younger groups (P < 0.005). Conversely, CBP night-time ABP difference did not vary among groups (24 +/- 20/16 +/- 11 mmHg). These age-dependent differences determined a rising prevalence of white-coat hypertension (from 19 to 40%, P = 0.001) and night/day ratio of at least 0.9 (from 43 to 66%, P = 0.0004). Age, diabetes, left ventricular hypertrophy and anemia but not GFR predicted nondipping status. Among the oldest patients, 13% had diastolic CBP below 70 mmHg, with 48% below the corresponding values of daytime (< 69 mmHg) or night-time ABP (< 60 mmHg). CONCLUSION: In CKD, prevalence of white-coat hypertension, nondipping status and potentially dangerous low diastolic ABP increases with aging. This suggests wider use of ABP monitoring in older patients and need for trials addressing identification of an age-specific blood pressure target. PMID- 17921830 TI - Reduction of blood pressure variability by combination therapy in spontaneously hypertensive rats. AB - OBJECTIVE: To test the hypothesis that combination therapy might be a powerful tool to reduce blood pressure variability (BPV) in the treatment of hypertension. METHODS: In chronic studies, male spontaneously hypertensive rats were given drugs in their rat chow for 18 weeks (n = 9 or 10 for each group of rats). Doses were as follows: atenolol (10 mg/kg per day), amlodipine (1 mg/kg per day), a combination of atenolol and amlodipine (2.5 + 0.25, 5 + 0.5, and 10 + 1 mg/kg per day), hydrochlorothiazide (8 mg/kg per day), enalapril (3.2 mg/kg per day), and a combination of hydrochlorothiazide and enalapril (2 + 0.8, 4 + 1.6, and 8 + 3.2 mg/kg per day). In acute studies, drugs were perfused through the left femoral vein in conscious rats. RESULTS: In chronic studies, compared with monotherapy, the combinations of two antihypertensive drugs were more effective in reducing the blood pressure (BP), BPV, and organ damage. The indexes of organ damage were all positively related to BP and/or BPV. In acute studies, the constant infusion of phenylephrine (6.25 microg/kg per min) markedly increased the BP, but showed no significant effects on BPV. The infusion of a combination of atenolol and amlodipine (62.5 + 6.25 microg/kg per min) or a combination of hydrochlorothiazide and enalapril (500 + 200 microg/kg per min) significantly reduced the BP and BPV; moreover, a significant reduction in BPV was still found when the rat's BP was restored to control levels by a concomitant infusion of phenylephrine. CONCLUSIONS: Combination therapy may be a powerful and useful tool for BPV reduction in the treatment of hypertension. In addition to the BP reduction, the decrease in BPV may contribute significantly to the prevention of organ damage in hypertension. PMID- 17921831 TI - Spironolactone has antiarrhythmic activity in ischaemic cardiac patients without cardiac failure. AB - OBJECTIVES: To examine whether endogenous aldosterone can cause either arrhythmias (and some of their underlying mechanisms) or endothelial dysfunction in patients with coronary artery disease (CAD) but without heart failure. BACKGROUND: Aldosterone blockade has been shown to reduce the incidence of sudden death in patients with heart failure. This could be caused by a reduction in arrhythmias or in coronary events. Whether either effect also occurs in other cardiac patients without heart failure is currently unknown. METHOD: We performed a randomized, placebo-controlled, double-blind crossover study on 98 patients with CAD but without heart failure on standard therapy, comparing 12.5-50 mg/day spironolactone (3 months) with placebo. Endothelial function was assessed by bilateral forearm venous occlusion plethysmography. Ventricular extrasystoles, procollagen III N-terminal peptide (PIIINP) and QT interval length were used to represent arrhythmias and their determinants. RESULTS: Spironolactone produced a highly significant 75% reduction in ventricular extrasystoles (median 192, range 48-744) on placebo compared with spironolactone (median 48, range 19.2-288, P < 0.003). Spironolactone also decreased the QT interval from a mean of 440 +/- 28 to a mean of 425 +/- 25 (P < 0.001) and a collagen marker (PIIINP) from a mean of 3.6 +/- 0.9 to a mean of 3.0 +/- 0.8 (P < 0.001), but did not significantly change endothelial dysfunction or heart rate variability. CONCLUSION: These results suggest that despite conventional therapy, endogenous aldosterone can be an arrhythmogenic influence in patients with CAD, but without heart failure. The possible mechanisms are that aldosterone promotes myocardial fibrosis and lengthens the QTc interval as well as decreasing potassium in CAD patients without heart failure. PMID- 17921832 TI - Crossover study of amlodipine versus nifedipine CR with home blood pressure monitoring via cellular phone: internet-mediated open-label crossover trial of calcium channel blockers for hypertension (i-TECHO trial). AB - OBJECTIVE: We developed a new data collection system named i-converter that could transmit data to a website via cellular phone. Using the system, we compared the effects of two calcium channel blockers on the home blood pressure. METHODS: Amlodipine and nifedipine CR were administered to 41 patients with essential hypertension for more than 6 weeks each in a randomized open-label crossover study. The dose of each drug was increased until the home blood pressure reached the target level of under 135/85 mmHg. RESULTS: The morning home systolic and diastolic blood pressures were significantly lower during nifedipine CR treatment: 133 +/- 10/81 +/- 8 mmHg with amlodipine versus 131 +/- 8/80 +/- 8 mmHg with nifedipine CR, P < 0.05. The morning pulse rate was significantly higher during nifedipine CR treatment (69 +/- 9 beats/min with amlodipine versus 70 +/- 9 beats/min with nifedipine CR, P < 0.05). The evening home blood pressure and pulse rate, however, showed no significant differences between the two drugs (128 +/- 11/74 +/- 7 mmHg and 74 +/- 10 beats/min with amlodipine versus 128 +/- 10/75 +/- 7 mmHg and 74 +/- 9 beats/min with nifedipine CR, all not significant). CONCLUSIONS: Nifedipine CR had a stronger antihypertensive effect than amlodipine during the critical morning period, but the morning pulse rate was higher. Our new data transmission system was effective for collecting precise data on the blood pressure and pulse rate via the internet. PMID- 17921833 TI - Minimally invasive transiliac plate osteosynthesis for type C injuries of the pelvic ring: a clinical and radiological follow-up. AB - OBJECTIVE: To evaluate radiological and functional outcome in patients treated with minimally invasive transiliac plate osteosynthesis for unstable pelvic injuries. DESIGN: Retrospective analysis of a prospective treatment protocol in a consecutive patient series. SETTING: Level 1 trauma center. PATIENTS: Between January 1998 and December 2005, 31 patients with type C injuries of the pelvic ring were treated with minimally invasive transiliac plate osteosynthesis. According to the AO classification, 16 patients had a C1-injury, 9 had a C2 fracture, and 6 patients sustained a C3 injury of the pelvic ring. Anterior posterior, inlet, and outlet radiographs were obtained preoperatively, immediately postoperatively, and during follow-up. Clinical outcome was determined according to the Hannover pelvic outcome score. INTERVENTION: Posterior plate osteosynthesis for type C injuries of the pelvic ring. MAIN OUTCOME MEASUREMENT: Preoperative and postoperative dislocation of the posterior pelvic ring, loss of reduction, implant failure, implant removal, clinical results of the pelvic injury and general limitations following the trauma. RESULTS: Maximum average dislocation of the posterior pelvic ring was 16.1 mm preoperatively; postoperatively, it was 6.1 mm. A total of 23 patients (74.2%) could be followed up after an average of 20 months (range 7-57 months). Seven patients underwent follow-up treatment at other hospitals closer to their respective residences, whereas 1 patient passed away in the early postoperative phase due to multiorgan failure. Loss of reduction occurred in 2 cases. The clinical outcome regarding the pelvis was very good in 8 cases, good in 9 cases, fair in 4 cases, and poor in 2 cases. Social reintegration according to the Hannover pelvic outcome score was complete in 9 cases, poor in 10 cases, and incomplete in 10 cases. CONCLUSION: Posterior plate osteosynthesis is a sufficiently stable method for the treatment of unstable pelvic ring injuries with a low risk of iatrogenic nervous tissue and vascular lesions. The disadvantages are limited reduction possibilities, the necessity of bilateral bridging of the sacroiliac joint in a unilateral injury, as well as a higher rate of symptomatic hardware. PMID- 17921834 TI - Predicting blood loss in isolated pelvic and acetabular high-energy trauma. AB - OBJECTIVE: To quantify transfusion requirements in patients with isolated acetabular or pelvic fractures and correlate these requirements with fracture classification. DESIGN: Retrospective review of 382 patients with isolated pelvic and/or acetabular fractures. SETTING: Academic Level I Trauma Center. PATIENTS/PARTICIPANTS: Patients were identified from a trauma registry. Appropriate radiographs and complete transfusion data were obtained for 289 (75%) of 382 eligible patients between January 1, 1998 and December 31, 2003. INTERVENTION: Classification of pelvic fracture by Young and Burgess type and acetabular fractures by Letournel type. MAIN OUTCOME MEASUREMENT: Number of units of blood transfused in the first 24 hours after admission to the trauma center. RESULTS: Patients with isolated pelvic fractures with major ligament disruption (APC II or III, LC III, vertical shear, or combined mechanisms) were more likely to receive a blood transfusion (44%) than other fracture types (8.5 %) (P < 0.0005). Transfusion amounts were greatest in APC III (12.6 units) and vertical shear (4.6 units) injuries. Fractures classified as both column, anterior column, anterior column posterior hemi-transverse, or T type were more likely to receive a blood transfusion (56%) than other fracture types (28%) (P = 0.003). Of these fracture types, both column (8.8 units) and anterior column posterior hemi transverse (6.4 units) received the largest transfusions. CONCLUSIONS: Patients with isolated acetabular fractures are as likely as those with isolated pelvic fractures to receive blood transfusions within the first 24 hours of admission. Higher energy pelvic ring fractures classified as APC II or III, LC III, vertical shear, or combined mechanism require more frequent transfusion than other pelvic fractures. Acetabular fractures involving the anterior column as well as T-type fractures require more frequent blood transfusions than other acetabular fractures. PMID- 17921835 TI - Augmentation of posterior wall acetabular fracture fixation using calcium phosphate cement: a biomechanical analysis. AB - OBJECTIVE: This study investigates if the use of calcium phosphate cement as an adjunct to internal fixation for posterior wall acetabular fracture will result in acute restoration of joint loading parameters to the intact condition. METHODS: Ten fresh-frozen cadaveric pelves were used for this investigation. Simulated abductor mechanism was used to load the hip. Pressure-sensitive film was used to measure contact area and pressure within the anterior, superior, and posterior regions of the acetabulum for all experimental conditions. The hips were loaded under the following 4 conditions: 1) intact; 2) following posterior wall osteotomy; 3) following reduction and standard internal fixation; and 4) following reduction of the posterior wall using calcium phosphate cement, as a grout, in addition to internal fixation. A posterior wall fracture was created along an arc of 40-90 degrees about the acetabular rim. Extensometers were utilized to measure posterior wall fragment micromotion under conditions 3 and 4 above. Statistical analysis was performed using multivariate analysis of variance to assess the significance of the difference among and between conditions simultaneously for each region. Fragment motion data were analyzed using a 2 tailed t test. RESULTS: Fragment micromotion was reduced to 78 microm superiorly and 46 microm inferiorly with the use of calcium phosphate cement (P < 0.05). Creation of a posterior wall defect resulted in increased load in the superior acetabulum (1201N) as compared to the intact condition (902N, P = 0.024). Following reduction and internal fixation, the load distributed to the superior acetabulum (1132N) was not statistically different from the displaced condition. Following the addition of calcium phosphate cement, the load seen at the superior region of the acetabulum (883N) was less than fixation without calcium phosphate cement and was not different from the intact state (P = 0.85). CONCLUSION: The use of calcium-phosphate cement as a fracture grout with internal fixation resulted in a partial restoration of joint loading parameters toward the intact state. Further work will be needed to determine if similar types of augmented articular fixation may result in a clinical benefit. PMID- 17921836 TI - Is there a gluteus medius tendon injury during reaming through a modified medial trochanteric portal? A cadaver study. AB - OBJECTIVES: The purpose of this cadaveric study was to evaluate whether there is any damage to the gluteus medius tendon when reaming through a modified medial trochanteric portal for antegrade intramedullary femoral nailing. METHODS: Ten cadaver hips were used in this study. A guidewire was placed in the modified medial trochanteric portal using the assistance of C-arm fluoroscopy and a 14-mm reamer was advanced over the wire. After the reaming was complete, each hip was dissected and the gluteus medius muscle and tendon were inspected to evaluate the amount of intrasubstance and medial tendon damage. RESULTS: Precise localization of the modified medial trochanteric portal was achieved in 9 of 10 cadaver hips. Of those nine hips, the use of the modified medial trochanteric portal did not result in any visible damage to the tendinous insertion of the gluteus medius or the medial aspect of the tendon in any of the specimens. CONCLUSIONS: There is no damage to the gluteus medius tendon with the use of the modified medial trochanteric portal. Although the clinical implications of this finding are not known with certainty, the use of the modified medial trochanteric entry portal for antegrade femoral nailing could possibly result in less postoperative morbidity because it does not damage the gluteus medius tendon as compared to the traditional more lateral trochanteric portal. PMID- 17921837 TI - Minimally invasive application of the non-contact-bridging (NCB) plate to the proximal humerus: an anatomical study. AB - OBJECTIVES: To describe a minimal anterolateral acromial approach for minimally invasive (MI) treatment of fractures of the proximal humerus (PH) with the Non Contact-Bridging (NCB) plate. DESIGN: 1) Cadaver study and 2) clinical case series. SETTING: 1) University Institute of Anatomy and the 2) University Level I trauma center. SPECIMENS/PATIENTS: 1) Ten fresh frozen human humeri and 2) 22 patients with 22 isolated proximal humeral fractures. INTERVENTION: 1) Minimal anterolateral acromial approach with MI application of the NCB-PH plate followed by dissection of the axillary nerve and 2) MI fracture fixation using this approach and technique of plate insertion. MAIN OUTCOME MEASURES: 1) Integrity of the axillary nerve and evaluation of its relationship to the implant, and 2) early postoperative functional results. RESULTS: In the cadaver study, the nerve directly crossed over the percutaneously inserted plate in all the arms. The nerve then divided into two branches anterior to the plate in eight arms and divided into two branches directly over the plate in two arms. One branch of the axillary nerve in one arm was injured. In the clinical case series, no intraoperative complications relating to the approach or the implant occurred. No symptoms of axillary nerve lesion have been detected so far in the early follow up. CONCLUSIONS: The minimal anterolateral acromial approach is suitable for MI technique to apply the NCB-PH. The relationship of the axillary nerve to the plate is anatomically close. We recommend that strict bone contact be maintained during plate insertion and that screw insertion complies with the guidelines provided for this technique. In a small clinical cases series, the plate and screws were inserted in accordance with these guidelines and no axillary nerve lesions have yet been detected. PMID- 17921838 TI - Minimally invasive plating osteosynthesis (MIPO) of middle and distal third humeral shaft fractures. AB - OBJECTIVE: To evaluate the clinical, radiographic, and functional outcomes of middle and distal third humeral shaft fractures treated with the minimally invasive percutaneous osteosynthesis (MIPO). DESIGN: Prospective, single-center, nonconsecutive clinical series study. SETTING: : Skeletal trauma center of a university teaching hospital. PATIENTS: Thirteen patients who were seen from May 2004 to October 2005 with an average age of 38.1 years (range, 25 to 60 years) form the basis of this study. Patients were obtained from a surgical database of 1 surgeon. INTERVENTION: The middle and the distal third humeral shaft fractures were reduced by closed means and fixed with long narrow 4.5-mm dynamic compression plates introduced through 2 small incisions away from the fracture sites and placed on the anterior aspect of the humerus. MAIN OUTCOME MEASUREMENTS: Time to fracture healing and functional assessments were assessed at an average follow-up of 12.5 months (range, 7 to 19 months) for the affected shoulders and elbows using the UCLA and Mayo elbow performance scoring systems, respectively. RESULTS: All fractures united with a mean healing time of 16.2 weeks (range, 12 to 32 years). There were no nonunions, radial nerve palsies, or implant failures. The UCLA scoring system showed excellent results in 7 cases (53.8%) and good results in 6 cases (46.2%). Thirteen patients had excellent results of their elbow function when assessed with the Mayo elbow performance scoring system. CONCLUSION: Closed reduction and internal fixation of middle or distal third humeral shaft fractures using MIPO is a safe and effective surgical treatment method and an alternative option to open techniques. PMID- 17921839 TI - Outcomes of tibial nonunion in older adults following treatment using the Ilizarov method. AB - OBJECTIVES: To describe the functional outcomes of treatment using the Ilizarov method for tibial nonunions in older patients (>60 years of age). DESIGN: Prospective case series. SETTING: Tertiary referral center. PATIENTS: Twenty three consecutive patients with an average age of 72 years (61 to 92) who had tibial nonunions for an average duration of 13 months (3 to 46). Fourteen patients had an associated deformity and eight patients had infection. INTERVENTION: Ilizarov deformity correction, compression, or bone transport. MAIN OUTCOME MEASUREMENTS: Brief Pain Inventory, American Academy of Orthopaedic Surgeons (AAOS) Lower Limb Core Scale, Short Form (SF)-12, quality-adjusted life years. RESULTS: Three patients did not complete treatment: two patients died of cardiovascular disease during the treatment period and one patient demanded early removal of the Ilizarov device against medical advice. All 20 patients who completed treatment achieved bony union. Two of the 20 patients died before final follow-up, one patient was unable to participate in follow-up, and one patient was lost. At an average follow-up of 38 months (18 to 61), all of the remaining 16 patients were bearing full weight. AAOS Lower Limb Core Scale scores improved from 39 to 78 points (P < 0.001), pain intensity decreased from 3.6 to 0.9 (P = 0.001), SF-12 Physical Component Summary scores improved from 26.5 points to 35.3 points (P = 0.030), and SF-12 Mental Component Summary scores improved from 41.6 points to 48.7 points (P = 0.011). The improvement in quality of life is equivalent to 5.3 quality-adjusted life years per patient, which was larger than the average improvement in quality of life following total hip arthroplasty reported in published series. CONCLUSIONS: Treatment using the Ilizarov method restored function and had a profoundly positive effect on quality of life in these elderly patients with tibial nonunions. PMID- 17921840 TI - Syndesmotic instability in Weber B ankle fractures: a clinical evaluation. AB - OBJECTIVE: Syndesmotic instability may coexist with unstable Weber B supination external rotation (SE) lateral malleolar fractures. Current recommendations suggest that Weber B injuries should not have associated syndesmotic instability after open reduction and internal fixation of the lateral malleolus. The purpose of this study was to evaluate syndesmotic stability with respect to the current recommendations for syndesmotic fixation in Weber B SE pattern lateral malleolar fractures. DESIGN: Retrospective cohort, consecutive series. SETTING: Academic Level I trauma center. PATIENTS/PARTICIPANTS: Over a 7-year period, 238 skeletally mature patients with unstable SE pattern Weber B lateral malleolus fractures with deltoid ligament incompetence were evaluated. INTERVENTION: After lateral malleolar fixation, syndesmotic stability was evaluated by an external rotation stress examination under direct vision and fluoroscopy. MAIN OUTCOME MEASURES: The incidence of syndesmotic instability as defined by previously reported criteria. RESULTS: Syndesmotic instability was found in 93 of the 238 (39%) fractures after fixation. Instability was identified in the operating room in 92 of the 93 ankles. One case of instability was missed intraoperatively and diagnosed 2 weeks after surgery. All other patients were followed to union without displacement. CONCLUSIONS: We found syndesmotic instability to be common after anatomic and stable bony fixation in unstable Weber B SE pattern lateral malleolar fractures. Previously published criteria for syndesmotic instability based on cadaveric studies are not representative of the clinical situation. Syndesmotic instability in conjunction with unstable Weber B SE pattern lateral malleolar fractures must be sought out in the operating room with an intraoperative stress examination. PMID- 17921841 TI - Table-skeletal fixation as an adjunct to pelvic ring reduction. AB - The reduction of displaced pelvic ring injuries remains a technical challenge, especially when treatment is delayed. A pelvic frame (Orthopaedic Systems Inc, Union City, California) provides a means of external skeletal fixation, rigidly stabilizing the intact hemipelvis to the operating room table. The fractured and displaced fragments can then be manipulated around the securely fixed uninjured hemipelvis, allowing the application of more directions and magnitudes of force for reduction maneuvers than allowed by the traditional means of pelvic reduction. The surgical technique and 1 case each of an acute fracture and pelvic nonunion/malunion are presented. PMID- 17921842 TI - The extended deltoid-splitting approach to the proximal humerus. AB - The recent technological developments in implant design and the wider availability of bone graft substitutes have stimulated a renaissance in the operative treatment of complex proximal humeral fractures. However, one of the remaining problems of the operative treatment of these injuries has been the limited surgical access to the posterior aspect of the shoulder afforded by the deltopectoral approach. In this article, we describe a novel extended deltoid splitting approach, in which the area traversed by the axillary nerve is identified and protected during the surgery. We feel that this approach provides enhanced surgical exposure and offers a useful alternative to the deltopectoral approach in the operative treatment of 3- and 4-part proximal humeral fractures. PMID- 17921843 TI - Extraction of intramedullary nails by proximal stacked wire technique. AB - We describe the use of proximal stacked wires for the removal of an intramedullary nail in the context of difficult extraction and failed attempts by conventional methods. This percutaneous technique is particularly useful for the extraction of narrow cannulated or solid intramedullary nails after failure of removal by extraction bolt devices. PMID- 17921844 TI - Plastic deformation and impaction of the retroacetabular surface associated with posterior fracture-dislocation of the hip: description of two cases. AB - In two cases of posterior fracture-dislocation of the hip, we found an impaction and plastic deformation of the retroacetabular surface, in addition to other typically associated lesions. This deformation creates a mismatch between the stable and the free fragments of the posterior wall, and if unrecognized makes an anatomic reconstruction of the posterior wall impossible. On the computed tomography scan, the most characteristic sign is a concave deformation of the retroacetabular region as compared to the contralateral side. To our knowledge, this specific fracture-dislocation pattern has not been previously described in the literature. If orthopedic surgeons know about the existence of this deformation, then they will easily recognize the impaction on preoperative computed tomography scan and will be able to achieve an anatomic reduction of the acetabulum. Diagnostic aspects and the operative technique to approach this problem are described and illustrated in two cases. PMID- 17921846 TI - A predictive method for subsequent avascular necrosis of the femoral head (AVNFH) by observation of bleeding from the cannulated screw used for fixation of intracapsular femoral neck fractures. PMID- 17921847 TI - Medulloblastomas expressing IL13Ralpha2 are targets for IL13-zetakine+ cytolytic T cells. AB - Central nervous system loco-regional disease relapse is a common etiology of treatment failure for medulloblastoma (MB)/primitive neuroectodermal tumors. Therapeutic targeting of primary disease and the adjacent craniospinal cerebral spinal fluid pathways should decrease relapse rates and allow for the curtailed use of radiation therapy. The adoptive transfer of tumor-specific cytolytic T cells (CTLs) to the tumor bed and cerebral spinal fluid is an attractive strategy, but limited in its clinical application owing to the paucity of defined antigens consistently expressed by these tumors and their potential to escape T cell recognition by expressing low level surface human leukocyte antigen. Here, we describe the human leukocyte antigen-independent recognition of MB cell surface IL13Ralpha2 by genetically modified CTLs expressing an IL13-zetakine chimeric immunoreceptor. We found that IL13-zetakine+ CTLs exhibit potent cytolytic activity toward IL13Ralpha2 Daoy cells, and are activated to secrete proinflammatory cytokines such as interferon-gamma. By employing an orthotopic NOD-scid murine model in which intraventricularly seeded Daoy cells form tumors on leptomeningeal surfaces, regression of established ffLuc+ Daoy xenografts in response to intraventricularly delivered IL13-zetakine+ CD8+ CTLs was observed using biophotonic imaging. These studies support the rationale for exploring the clinical utility of targeted immunotherapy using adoptively transferred IL13 zetakine redirected CTLs as a therapeutic component for treating IL13Ralpha2+ MB/primitive neuroectodermal tumors. PMID- 17921848 TI - Extracorporeal photopheresis for steroid resistant graft versus host disease in pediatric patients: a pilot single institution report. AB - This study was aimed at ascertaining whether extracorporeal photopheresis (ECP) is an effective treatment for pediatric patients with steroid resistant graft versus host disease (GvHD). Fifteen patients with acute GvHD (aGvHD) and 10 patients with chronic GvHD (cGvHD) were enrolled in the study. At the start of the ECP protocol, aGvHD was staged as II (n=7), III (n=4), and IV (n=4). The response rate was 100% for aGvHD II, 75% for aGvHD III, and finally 0% for aGvHD IV (P=0.02). In multivariate analysis, the strongest predictor for ECP response was the aGvHD severity: aGvHD II 100%, aGvHD III-IV 30% [relative risk (RR) 5.071, confidence interval (CI) 95% 2.2-5.5, P=0.0016], this translates in a higher risk of transplant-related mortality for ECP nonresponders (RR 5.26, CI 95% 3.4-6.2, P=0.02). cGvHD was diagnosed as limited n=3, and extensive n=7; the response rate was 100% and 28% for limited or extensive cGvHD, respectively (P=0.03). For cGvHD the strongest predictor for ECP response was the absence of visceral organ involvement (RR 5.17, CI 95% 2-4.9, P=0.001), and the highest risk of transplant-related mortality was among patients not responding to ECP (RR 12.4, CI 95%, P=0.02). Our results suggest that ECP can rescue good-risk GvHD patients, whereas for advanced, poor-risk GvHD patients, new therapies are required. PMID- 17921849 TI - Retrospective study on elimination delay of methotrexate in high-dose therapy of childhood acute lymphoblastic leukemia in China. AB - OBJECTIVES: The aim of this study was to observe the morbidity of elimination delay in Chinese children with acute lymphoblastic leukemia during high-dose methotrexate (HDMTX) therapy and the toxicities. PATIENTS AND METHODS: A total of 121 children with acute lymphoblastic leukemia on HDMTX therapy were enrolled into this study. Patients were divided into groups on the basis of either dosage (3 g/m vs. 5 g/m) or infusion duration (7 h vs. 24 h). CF/MTX index was used to determine the calcium folinate (CF) rescuing intensity and toxicity was evaluated according to World Health Organization criteria. RESULTS: The overall morbidity of elimination delay was 12.1% in a total of 497 infusions. Patients with elimination delay had lower platelet count (P<0.01) and greater cumulative CF rescuing intensity (P<0.001). In 3-g group, children with elimination delay experienced severer oral mucous membrane damage (P<0.05) than those without elimination delay, and postponement of following chemotherapy (P=0.001). No significant difference was found in morbidity of elimination delay between 3 and 5-g groups (P>0.05) or 7 and 24-hour infusion groups (P>0.05). The only raised adverse effect in 5-g group was gastrointestinal (P=0.003) as compared with 3-g group. The CF rescuing intensity of 5-g group without elimination delay was lower than that of the 3-g group (P<0.01). CONCLUSIONS: (1) HDMTX with 5 g/m is as safe as 3 g/m under adequate hydration and alkalization. Twenty-four-hour infusion is optimal. (2) Individualized dosing is necessary. PMID- 17921850 TI - Does the periodontal health of thalassemia major patients have an impact on the blood lipid profiles? A preliminary report. AB - Cardiac symptoms and premature death from cardiac causes are still suggested to be a major problem in thalassemic patients. One of the main reasons for the cardiovascular events in thalassemia major (TM) patients has suggested having iron overload, in addition to other reasons such as hypoxia, abnormal lipoproteins levels, atherosclerotic conditions, etc. This study aims to investigate whether the periodontal health has an impact on the blood lipid levels. Twenty-four TM patients and sex-matched controls (C) enrolled to this study. The measurements of the periodontal parameters (gingival index, plaque index, bleeding on probing, probing depth, and clinical attachment level) were recorded. Besides, in venous blood samples the lipid profile was investigated. All of the periodontal parameters were significantly higher in TM group than in C group (P<0.05). Triglyceride and cholesterol/high density lipoprotein ratio had significant positive correlations with the periodontal parameters (P<0.05). Multiple regression analyses present significant associations between gingival index, plaque index scores, and the cholesterol/high density lipoprotein ratio (P<0.001). Our study results showed that the lipid profile of TM patients might be affected by their periodontal health. Further studies are needed to determine the lipid profile involvement magnitude and the cardiovascular disease risk caused by the periodontal health of TM patients. PMID- 17921851 TI - The addition of ifosfamide/etoposide to cisplatin/teniposide improves the survival of children with retinoblastoma and orbital involvement. AB - This study aimed to determine the impact of the addition of ifosfamide/etoposide to a regimen containing cisplatin/teniposide on the survival of patients with retinoblastoma with orbital involvement. Thirty patients were treated at the A. C. Camargo Hospital, Brazil, from 1986 to 2002. From 1986 to April 1992 (period I, n=12), treatment consisted of 3 cycles of induction chemotherapy with cisplatin and teniposide, followed by maintenance with same drugs alternating with cyclophosphamide, vincristine, and doxorubicin every 21 days for 60 weeks. Since April 1992 (period II, n=18), the treatment consisted of 3 cycles of ifosfamide and etoposide followed by maintenance with same drugs, alternating with cisplatin and teniposide every 21 days for 36 weeks. In both periods, children were submitted to exenteration with eyelid preservation and orbital radiation therapy with 45 cGy, and also received intrathecal therapy with methotrexate plus dexamethasone and cytarabine. Kaplan-Meier method was used for survival analysis. The median age was 31 months. Most patients (86.7%) presented unilateral tumors. The 3-year overall survival was 34.4% and 72.2%, respectively, for patients treated during periods I and II (P=0.061). The addition of ifosfamide/etoposide to chemotherapy with cisplatin/teniposide improves survival in these patients, but further studies are still necessary. PMID- 17921852 TI - Factors influencing complementary and alternative medicine use in a multisite pediatric oncology practice. AB - PURPOSE: Complementary and alternative medicine (CAM) use in children is common although estimates of prevalence vary widely. We studied CAM use in our multisite pediatric oncology practice and evaluated factors influencing CAM use. PATIENTS AND METHODS: We conducted an anonymous cross-sectional survey of 274 parents of children treated at the combined Nemours oncology practice in Florida and Delaware. Prevalence of CAM use was determined and binary logistic regression was used to evaluate factors related to CAM use in children. RESULTS: The prevalence of CAM use among children and parents was 24.5% and 66.7%, respectively. Intensity of parent's use of CAM and geographic region were significantly associated with CAM use in children. The odds of CAM use in children increased with increasing use among parents. When parents used 6 or more therapies children were 33 times more likely to use CAM compared with those whose parents did not use CAM (odds ratio=33.3; 95% confidence interval, 10.4-106.2, P<0.01). Children in Florida were more likely to use CAM compared with children in Delaware (odds ratio=3.0; 95% confidence interval, 1.6-5.8, P<0.01). CONCLUSIONS: These results indicate that children's use of CAM is significantly related to the intensity of parent's use regardless of parent's race, sex, education, household income, or child's sex or age. Clinicians should consider parental use and intensity of CAM use. Assessing CAM use should include classifications established by the National Center for CAM and a standard format for inquiring about CAM use should be developed. PMID- 17921854 TI - T-cell acute lymphoblastic leukemia in a child with ataxia-telangiectasia: case report. AB - We present a patient with acute lymphoblastic leukemia and ataxia-telangiectasia (A-T). The 4-year-old girl is the first child of young nonconsanguineous parents of Serbian origin. Gait problems appearing in the second year of life were treated by physiotherapy. At the age of 4 she was diagnosed with T-cell acute lymphoblastic leukemia and treated according to Berlin-Frankfurt-Munster strategy. Owing to typhlitis developing after 15 days of cytotoxic treatment, frequent radiologic examinations were performed causing profound aplasia. Typhlitis did not respond to conservative treatment but necessitated extensive bowel resection. At that time the A-T was suspected by our team and confirmed by increased chromosomal radiosensitivity and markedly reduced level of A-T mutated protein. Chemotherapy was continued without alkylating agents and further radiologic imaging ran an uncomplicated course. At present, the patient is in first remission and 2.5 years since the beginning of the treatment. We stress the importance of careful initial neurologic evaluation of children with malignancy. PMID- 17921853 TI - Human herpesvirus 6 (HHV-6)-associated pleurisy after unrelated cord blood transplantation in children with chemotherapy-resistant malignant Lymphoma. AB - Two children, 5 and 10 years of age, received unrelated cord blood transplantation (CBT) for malignant lymphoma. Both of them suffered from pleurisy with and without interstitial pneumonitis after transplantation. By the quantitative real-time polymerase chain reaction, human herpesvirus 6 (HHV-6) variant B DNA was detected in pleural effusion. This is the first report of HHV-6 associated pleurisy after hematopoietic stem cell transplantation. HHV-6 associated pleurisy should be considered as a complication after hematopoietic stem cell transplantation even in the absence of pneumonitis. Quantitative polymerase chain reaction is a useful tool for rapid detection of viral DNA, which may facilitate precise diagnosis and appropriate treatment. PMID- 17921855 TI - A case of recurrent paroxysmal cold hemoglobinuria with the different temperature thresholds of Donath-Landsteiner antibodies. AB - In this article, we first report a case of recurrent paroxysmal cold hemoglobinuria with serologic confirmation. On 2 occasions, the Donath Landsteiner (DL) antibodies belonged to an IgM subclass and showed neither anti-P nor anti-I specificity. Furthermore, it is very interesting that the temperature thresholds of DL antibodies were different on each occasion. Although acute paroxysmal cold hemoglobinuria is considered to be self-limited and transient, we should be careful of its possible recurrence. DL tests must be repeated after the complete recovery from the first episode, with careful attention to several possible causes of false-negative DL tests. PMID- 17921856 TI - Acute hepatic sequestration associated with pneumococcal infection in a 5-year old Boy with sickle beta degrees -thalassemia: a case report and review of the literature. AB - A 5-year-old black male with sickle beta degrees -thalassemia presented with fever and a vaso-occlusive crisis. Within hours, he developed progressive hepatomegaly with an acute drop in the hemoglobin level that was refractory to repeated red blood cell transfusion. His condition deteriorated and eventually he succumbed to cardiorespiratory failure related to sepsis. Blood cultures grew Streptococcus pneumoniae. To the best of our knowledge, this is the first reported case of hepatic sequestration in a young child and the first reported in a patient with sickle beta degrees -thalassemia. We describe here the clinicohematologic and pathologic features of this case consistent with acute hepatic sequestration and present a review of the relevant literature. PMID- 17921857 TI - Transformation of childhood MDS-refractory anemia to acute lymphoblastic leukemia. AB - Myelodysplastic syndromes (MDS) are clonal disorders of hematopoietic stem cell. Patients have a deteriorating course with about 30% evolving into acute leukemias usually of the myeloid phenotype. Evolution into acute lymphoblastic leukemia is a rare and intriguing phenomenon seen in far less than 1% of adult cases, and extremely rare in pediatric population. We report a case of childhood MDS refractory anemia transforming into acute lymphoblastic leukemia after an interval of 21 months since presentation and being on cyclosporine therapy for 9.5 months. The case raises further questions about the biology of MDS and the potential role of cyclosporine in leukemic transformation. PMID- 17921858 TI - Successful treatment of Diamond Blackfan anemia with metoclopramide. PMID- 17921859 TI - Regenerative arrest of inflamed peripheral nerves: role of nitric oxide. AB - Inflammation can both support and hinder regeneration. In this work, we asked whether regeneration of peripheral nerve axons is facilitated or interrupted when it proceeds through a zone of local but nondirected inflammation. Regeneration was examined in new nerve bridges forming through conduits connecting transected rat sciatic nerves. The conduits, infused with lipopolysaccharide to generate a sterile and nondirected inflammatory response, had substantial rises in inducible nitric oxide (iNOS) mRNA synthesis. iNOS was expressed within macrophages just beyond the zone of axon regrowth. Under these conditions, there was complete interruption of regenerative bridge formation in all instances without axon regrowth across the transection. In a separate cohort, infusion of a broad spectrum NOS inhibitor (Nomega-nitro-L-arginine-methyl ester) into the conduit salvaged bridge formation in a proportion (3/8) of rats. Our findings indicate that local inflammatory conditions inhibit regenerative events and that nitric oxide may contribute to these events. PMID- 17921860 TI - The expression and distribution of neural salient serine/arginine-rich protein 1 in rat retina. AB - Neural salient serine/arginine-rich protein 1 (NSSR1) expression has been found in mouse cerebral neurons, cerebellar Purkinje cells, pyramidal neurons and granule cells of dentate gyrus and regulates the pre-mRNA splicing of genes important for neural functions. In this study, we demonstrated that NSSR1 is expressed in rat retina and extensively distributed in the outer and inner plexiform layers. Double staining experiments showed that NSSR1 distributed mainly in ON-type bipolar cells and localized in the dendrites, somata and axon terminals. The result suggests that NSSR1 may play important roles in retinal function, possibly via regulating the neural-specific alternative splicing of genes. PMID- 17921862 TI - Neural correlates underlying perception of tonality-related emotional contents. AB - Using an event-related functional MRI technique, we examined the blood oxygen level-dependent responses of normal participants to auditory stimuli that consisted of four triads to explore the neural correlates for judging mode related emotional contents in tonal music. Three categories of stimuli, MAJOR, MINOR and NEUTRAL were prepared. MAJOR and MINOR stimuli suggest C major and c minor, respectively. NEUTRAL stimuli were controls. The task was to judge the categories. Contrasts MAJOR-NEUTRAL and MINOR-NEUTRAL showed significant activation in the bilateral inferior frontal gyri, medial thalamus, and dorsal anterior cingulate cortex. It is suggested that the bilateral inferior frontal gyri and medial thalamus are involved in judging the mode, whereas the dorsal anterior cingulate cortex was related to conflicts in the participant's mind. PMID- 17921861 TI - Guidance cues from the embryonic dorsal spinal cord chemoattract dorsal root ganglion axons. AB - In the early stages, the dorsal root ganglion neurons extend their axons toward the dorsal spinal cord. We previously showed that surround repulsion by semaphorin 3A prevents sensory axons from straying from their paths. The finding, however, that sensory trajectories toward the dorsal spinal cord are almost normal in semaphorin 3A-deficient littermates raises the possibility that a chemoattraction-based mechanism also contributes to the formation of sensory axonal projections. By employing culture assays, we show that the dorsal spinal cord secretes chemoattractants for the dorsal root ganglion axons. Furthermore, we demonstrate that the activity of a dorsal spinal cord-derived cue is specific for early sensory axons. These results suggest that dorsal spinal cord-derived chemoattractants contribute to the formation of the initial trajectories of sensory axons. PMID- 17921864 TI - Antidepressant effect of stem cell-derived monoaminergic grafts. AB - In this study, we demonstrate that embryonic stem cells can be engineered to differentiate into high percentages of serotonergic and dopaminergic neurons. In vitro, these cells release serotonin and dopamine in response to membrane depolarization. Upon engraftment into the medial prefrontal cortex in rats, the homolog of the human anterior cingulate cortex, the cells assumed neuronal morphologies, expressed monoaminergic-specific proteins, and seemed to functionally integrate, as assessed by the upregulation of the immediate-early gene, cfos. Furthermore, the transplanted animals performed in a manner similar to that of animals that received the antidepressant, citalopram, when administered the forced swim test, a validated model of human depression. These results suggest that transplantation of customized stem cells might perhaps be useful in the study treatment of psychiatric disorders. PMID- 17921865 TI - Endosomal accumulation of GM1 ganglioside-bound amyloid beta-protein in neurons of aged monkey brains. AB - We performed an immunohistochemical analysis of the GM1 ganglioside-bound amyloid beta-protein (GAbeta), an endogenous seed of Alzheimer amyloids, in sections of cerebral cortices of cynomolgus monkeys of different ages from 4 to 36 years old. Here, we show that neuronal GAbeta immunostaining significantly increases in the sections obtained from animals at ages below 19 years, even without senile plaque formation, and that GAbeta accumulation exclusively occurs in organelles involved in the endocytic pathway, including early, late, and recycling endosomes, not in those involved in the secretory pathway. Together with previous findings that Abeta generation likely occurs in early endosomes and that GM1 accumulation in early endosomes is induced by endocytic pathway abnormalities, our results provide further evidence that endosomes are intimately involved in the Abeta associated pathology of Alzheimer's disease. PMID- 17921866 TI - Melatonin modulates glycine currents of retinal ganglion cells in rat. AB - Melatonin is a hormone participating in the modulation of various physiological functions via binding to specific melatonin receptors. In the retina, melatonin is synthesized and released by photoreceptors and may play a neuromodulatory role. By using patch clamp techniques, we demonstrate for the first time that glycine-induced currents from a population of isolated ganglion cells in the rat retina are potentiated by melatonin of nanomolar concentrations by increasing the efficacy and the channel conductance of the strychnine-sensitive glycine receptor. The melatonin effect is blocked by 4-P-PDOT, indicating the mediation of the MT2 receptor. These results suggest that melatonin, along with the MT2 receptor, may be involved in retinal information processing by modulating glycine receptor-mediated inhibition. PMID- 17921863 TI - Dp71, utrophin and beta-dystroglycan expression and distribution in PC12/L6 cell cocultures. AB - Function of dystrophin Dp71 isoforms is unknown but seems related to neurite outgrowth and synapse formation. To evaluate Dp71 role in myoneural synapses, we established a coculture model using PC12 cells and L6 myotubes and analyzed expression and localization of Dp71 and related proteins, utrophin and beta dystroglycan, in PC12 cells. Confocal microscopy showed Dp71d isoform in PC12 nuclei, golgi-complex-like and endoplasmic reticulum-like structures, whereas Dp71ab concentrates at neurite tips and cytoplasm, colocalizing with beta dystroglycan, utrophin, synaptophysin and acetylcholine receptors. Evidences suggest that Dp71ab isoform, unlike Dp71d, may take part in neurite-related processes. This is the first work on Dp and members of Dp-associated protein complex roles in a cell-line based coculturing system, which may be useful in determining Dp71 isoforms associations. PMID- 17921867 TI - Perceptual visual filling-in of toxoplasmic retinochoroiditis scotomas. AB - Perceptual visual filling-in of toxoplasmic retinochoroiditis chronic scotomas was studied in 13 patients. Although there is deficit of visual inputs, patients with such chronic lesions perceive the region in the visual field uninterrupted. Targets were programmed to appear just outside the edge of the retinal lesion, and healthy retinal areas in the same eye. The results showed no significant difference in perceptual filling-in latencies (P>0.05). CONCLUSIONS: (i) neural mechanisms of filling-in at the blind spot are probably involved explaining this perceptual phenomenon; (ii) any neuronal changes occurring at the edge of the lesion only affected filling-in within the scotoma area. PMID- 17921868 TI - The biological effects of cell-delivered brain-derived neurotrophic factor on cultured spiral ganglion cells. AB - The benefit achieved by the use of cochlear implants depends among other factors on the number of surviving spiral ganglion cells (SGCs). Neurotrophic factors, especially brain-derived neurotrophic factor (BDNF), have a protective effect on spiral ganglions. Coating of the cochlear implant electrode with BDNF-producing cells may provide long-term delivery of the factor. Therefore, the hypothesis that BDNF-producing fibroblasts can enhance cell survival of cultured SGCs was tested. Lentiviral infection of fibroblasts resulted in BDNF production. Conditioned medium obtained from infected fibroblasts was used for the cultivation of SGCs. As a result, improved survival and neurite outgrowth was observed on SGCs. Our results demonstrate that lentivirally infected fibroblasts produce BDNF that has neurotrophic effects on spiral ganglions. PMID- 17921869 TI - Involvement of the rat prefrontal cortex in a delayed reinforcement operant task. AB - To determine whether the rat medial prefrontal cortex (PFC) is involved in delayed reinforcement operant behavior, we studied the effects of transient inactivation of the medial PFC (or the hippocampus as a control) during a delayed reinforcement lever-press task. We demonstrated the involvement of the PFC in this task: PFC inactivation but not hippocampal inactivation significantly impaired performance. In a separate experiment, we also recorded the prefrontal multiple unit activities during the task to determine the roles of the PFC in detail. Neuronal activity decreased during the delay period, suggesting that this decrease plays a role in delayed reinforcement operant behavior. PMID- 17921870 TI - Auditory temporal resolution in children assessed by magnetoencephalography. AB - By means of magnetoencephalography we investigated the auditory-evoked fields (AEFs) elicited by broadband noise bursts in a gap-detection paradigm in children. AEFs of 16 healthy children (mean age 8.7 years) were recorded while they passively listened to 100-ms white-noise bursts with temporal gaps of 3, 6, 10 and 30 ms inserted after 5 or 50 ms. The peak of the earliest and largest positivity occurred at 97 ms and was evaluated by spatiotemporal source analysis. Psychophysical gap-detection thresholds were obtained for the same children. We found that the neuromagnetic gap responses corresponded to the psychoacoustic thresholds. AEFs thus provide an objective tool to assess auditory temporal resolution in children. Children's neuromagnetic response patterns differed significantly from the adult responses under the same experimental conditions. PMID- 17921871 TI - Automatic auditory change detection in humans is influenced by visual-auditory associative learning. AB - Automatic detection of auditory changes that violate a regular sound sequence is indexed by the mismatch negativity (MMN) component of the event-related potential. The MMN is considered to reflect an auditory sensory memory and attention switching mechanism. Our aim was to study whether the auditory MMN can be associated with visual cues that have predictive value. By using visual cues that predicted the appearance of a deviant sound in most but not all of the cases, we were able to elicit MMN not only to the deviant sounds but also to those regular sounds that were misleadingly preceded by the visual cue. This result indicates high flexibility in the human automatic auditory change detection system, as it is affected by short-term visual-auditory associative learning. PMID- 17921872 TI - Time-dependent changes in cortical excitability after prolonged visual deprivation. AB - Transcranial magnetic stimulation applied to the occipital cortex can elicit phosphenes. Changes in the phosphene threshold provide a measure of visual cortex excitability. Phosphene threshold was measured in participants blindfolded for five consecutive days to assess the effects of prolonged visual deprivation on visual cortical excitability. After 48 h of blindfolding, an acute decrease in phosphene threshold was observed, followed by a significant increase by day 5. Phosphene threshold returned to preblindfold levels within 2 h of light re exposure. Thus, light deprivation is characterized by a transient increase in visual cortical excitability, followed by a sustained decrease in visual cortex excitability that quickly returns to baseline levels after re-exposure to light. PMID- 17921873 TI - Concentrative meditation enhances preattentive processing: a mismatch negativity study. AB - The mismatch negativity (MMN) paradigm that is an indicator of preattentive processing was used to study the effects of concentrative meditation. Sudarshan Kriya Yoga meditation is a yogic exercise practiced in an ordered sequence beginning with breathing exercises, and ending with concentrative (Sahaj Samadhi) meditation. Auditory MMN waveforms were recorded at the beginning and after each of these practices for meditators, and equivalently after relaxation sessions for the nonmeditators. Overall meditators were found to have larger MMN amplitudes than nonmeditators. The meditators also exhibited significantly increased MMN amplitudes immediately after meditation suggesting transient state changes owing to meditation. The results indicate that concentrative meditation practice enhances preattentive perceptual processes, enabling better change detection in auditory sensory memory. PMID- 17921874 TI - Neurotrophic Schwann-cell factors induce neural differentiation of bone marrow stromal cells. AB - Neural transdifferentiation of bone marrow stromal cells has been questioned, because cell fusion could explain the development of new cell types, misinterpreted as transdifferentiated cells. We performed here cocultures of bone marrow stromal cells and Schwann cells, without possibility that both cell types can establish contact. In these conditions, bone marrow stromal cells expressed nestin 4 h after beginning cocultures, and strong expression of neuronal markers was disclosed at 72 h, increasing at 1 and 2 weeks. Our results support that neural transdifferentiation of bone marrow stromal cells is induced by soluble factors provided by glial cells, and suggest that cell fusion should not be significant when local bone marrow stromal cells administration for neural repair is considered. PMID- 17921875 TI - Hippocampal functional connectivity reflects verbal episodic memory network integrity. AB - Using functional magnetic resonance imaging during a verbal memory task, we investigated correlations of signal fluctuations within the hippocampus and ipsilateral frontal as well as temporal areas in temporal lobe epilepsy patients. Declarative memory abilities were additionally examined before and after temporal lobe epilepsy surgery. A significant difference exists in functional connectivity between patients whose mnemonic functions deteriorated and those who remained stable or improved. Univariate analyses showed significantly higher preoperative coupling between the hippocampus and Brodmann area 22 for the group that decreased in verbal learning. We suggest greater coupling to reflect higher functional network integrity. Postoperatively reduced learning ability in patients with higher preoperative coupling underlines the importance of hippocampal interaction with cortical areas for successful memory formation. PMID- 17921876 TI - Dopamine induces TNFalpha and TNF-R1 expression in SH-SY5Y human neuroblastoma cells. AB - Cytotoxic concentrations of dopamine (100-500 microM DA) induce expression of tumour necrosis factor receptor-1 (TNF-R1) and tumour necrosis factor-alpha (TNFalpha) in SH-SY5Y human neuroblastoma cells. TNFalpha expression is dose dependent and can also be detected after 6-hydroxydopamine (6-OHDA) or 1-methyl-4 phenylpyridinium iodide (MPP) treatment. The expression of TNF-R1 is also dose dependent, but was not observed in 6-OHDA or MPP-treatment. Cells not expressing TNF-R1 were insensitive to TNFalpha, whereas those treated with DA showed a further decrease in viability when subsequently treated with TNFalpha. Thus, DA treatment confers sensitivity to TNFalpha. The decrease of cell viability caused by DA was in part prevented by neutralizing TNFalpha with anti-TNFalpha. As TNF R1 is increased in substantia nigra of Parkinsonian brains, we suggest that nonvesiculated DA might also play a role in inducing TNF-R1 expression and predispose the neuron to the action of cytokines released in a microglia-mediated inflammatory response. PMID- 17921877 TI - Galanin plays a role in the conditioning lesion effect in sensory neurons. AB - Sensory neurons show enhanced neurite outgrowth in vivo and in vitro following a conditioning lesion. Previous studies have shown that these effects are dependent on two members of the gp130 family of cytokines, leukemia inhibitory factor and interleukin-6. Here, we asked whether galanin, a neuropeptide induced by these cytokines, plays a role in the conditioning lesion response. Following a conditioning lesion, neurite outgrowth in culture was reduced in sensory neurons from galanin -/- mice compared with those from wild type controls. In neurons from wild type mice, the length of the longest neurite was increased 2.4-fold after a conditioning lesion, compared with 1.8-fold in neurons from knockout animals. The results indicate that the induction of galanin plays an important role in triggering the conditioning lesion response. PMID- 17921878 TI - Development and organization of the human brain tissue compartments across the lifespan using diffusion tensor imaging. AB - We used a diffusion tensor imaging-based whole-brain tissue segmentation to characterize age-related changes in (a) whole-brain grey matter, white matter, and cerebrospinal fluid relative to intracranial volume and (b) the corresponding brain tissue microstructure using measures of diffusion tensor anisotropy and mean diffusivity. The sample, a healthy cohort of 119 right-handed males and females aged 7-68 years. Our results demonstrate that white matter and grey matter volumes and their corresponding diffusion tensor anisotropy and mean diffusivity follow nonlinear trajectories with advancing age. In contrast, cerebrospinal fluid volume increases linearly with age. PMID- 17921879 TI - Methamphetamine increases basal ganglia iron to levels observed in aging. AB - Increases in basal ganglia iron are well documented for neurodegenerative diseases but have not been associated with methamphetamine (METH). In this study, vervet monkeys that received two doses of METH (2 mg/kg, intramuscularly, 6 h apart) showed at 1 month, iron increases in substantia nigra pars reticulata and globus pallidus, with concurrent increases of ferritin-immunoreactivity and decreases of tyrosine hydroxylase-immunoreactivity in substantia nigra. At 1.5 years, substantia nigra tyrosine hydroxylase-immunoreactivity had recovered while iron and ferritin-immunoreactivity increases persisted. Globus pallidus and substantia nigra iron levels of the adult METH-exposed animals (age 5-9 years) were now comparable with those of drug-naive, aged animals (19-22 years), suggesting an aging-related condition that might render those regions more vulnerable to oxidative stress. PMID- 17921880 TI - Automatic detection of lexical change: an auditory event-related potential study. AB - We investigated the detection of rare task-irrelevant changes in the lexical status of speech stimuli. Participants performed a nonlinguistic task on word and pseudoword stimuli that occurred, in separate conditions, rarely or frequently. Task performance for pseudowords was deteriorated relative to words, suggesting unintentional lexical analysis. Furthermore, rare word and pseudoword changes had a similar effect on the event-related potentials, starting as early as 165 ms. This is the first demonstration of the automatic detection of change in lexical status that is not based on a co-occurring acoustic change. We propose that, following lexical analysis of the incoming stimuli, a mental representation of the lexical regularity is formed and used as a template against which lexical change can be detected. PMID- 17921881 TI - Proliferation of neural stem cells correlates with Wnt-3 protein in hypoxic ischemic neonate rats after hyperbaric oxygen therapy. AB - Hyperbaric oxygen therapy promoted brain cell proliferation. Wnt-3 is closely associated with the proliferation of neural stem cells. We examined whether hyperbaric oxygen promoted neural stem cells to proliferate and its correlation with Wnt-3 protein in hypoxic-ischemic neonate rats. Hyperbaric oxygen therapy was administered 3 h after hypoxia ischemia daily for 7 days. The proliferating stem cells and Wnt-3 protein were examined dynamically in the subventricular zone. Results showed that stem cells proliferated and peaked 7 days after hyperbaric oxygen therapy. Wnt-3 protein increased to the higher levels 3 days after therapy. Linear regression analysis showed that nestin protein correlated with Wnt-3 protein. We propose that hyperbaric oxygen treatment promote stem cells to proliferate, which is correlated with Wnt-3 protein. PMID- 17921884 TI - Looking at childhood obesity through the lens of Baumrind's parenting typologies. AB - Obesity is becoming the leading negative health outcome for the current generation of children to a greater degree than for any previous generation. Pediatric orthopaedic nurses encounter many patients and families with concerns about obesity and need the ability to promote parenting capacity in order to detect, prevent, or treat childhood obesity. Parenting is a complex process with numerous two-way interactions between the parent and child. Pediatric orthopaedic nurses affect parenting capacity daily as they care for families in all care settings. Many family researchers use Baumrind's parenting typologies (styles) and their correlations to child health outcomes in research. Understanding Baumrind's theories can help pediatric orthopaedic nurses understand the mechanisms parents use to affect the health outcomes related to the obesity of their children. Baumrind's is one parenting theory that can help demonstrate how parental behaviors and practices affect a child's self-concept and self-care development and ultimately a child's health promotion beliefs and practices related to obesity prevention and care that continue into adulthood. Nurses can use reviews of literature and application to practice of parenting styles to expand their repertoire of parent guidance and anticipatory teaching directed to the prevention and care of childhood obesity. PMID- 17921886 TI - Bone loss. An emerging problem following obesity surgery. AB - The incidence of obesity worldwide has increased markedly in the past 2 decades, with estimates of increases of 50% in the United States alone. Research indicates that weight loss produced by diet alone is not sustained and that 75% of dieters regain most of the weight lost within 1 year and 90% within 2 years. Morbid obesity is associated with comorbid conditions, including heart disease, hypertension, diabetes, mechanical arthropathy, sleep apnea, and numerous other serious disorders and a shortened life expectancy. Because of limited success with medical management, surgical treatment of morbid obesity has been used increasingly, especially with the development of laparoscopic procedures, including Roux-en-Y gastric bypass (RYGB). RYGB is associated with low surgical mortality, marked decreased food intake, and significant, sustained weight loss. However, in this emerging, unique population there is growing appreciation that these procedures may be associated with the development of bone loss and skeletal fragility because of altered nutrient metabolism. Despite the threat of skeletal fragility and fracture, there is limited data addressing the effects of bariatric surgery on bone metabolism and bone loss. PMID- 17921888 TI - Literature review on the effects of obesity on knee osteoarthritis. AB - Knee osteoarthritis is the most common form of arthritis. This has been partially noted because of the increase of the obesity trend in the United States. In addition it has been noted as a major cause of morbidity, mortality, and disability among the elderly. Although there have been several advances in this area, there are several areas that can be addressed by nurse clinicians. The effects of obesity, risk factors, and potential complications are discussed in this literature review. PMID- 17921889 TI - Screening for metabolic syndrome. PMID- 17921890 TI - Health and independence of young adults with disabilities. Two years later. AB - PURPOSE: American culture expects young people to complete school, find satisfying work, develop social networks, stay healthy, contribute to the maintenance and support of households, and participate as citizens in their communities. We know little about the life trajectories of young adults with disabilities during emerging adulthood. METHODS: Four-page surveys were mailed to the 650 respondents of a 10-page survey sent to graduates of a state Children with Special Healthcare Needs program and a children's orthopaedic hospital 2 years earlier to determine changes in healthcare access and use, insurance status, health perceptions, education, work, and markers of independent living over those 2 years of maturation. SAMPLE: The study sample was 300 young people who completed both surveys. Respondents were 20-25 years of age, 95% White, 64% female, and 28% who reported that they were independent in all activities of daily living (ADL). RESULTS AND DISCUSSION: Compared with the initial survey, results for this group of young people who reported needing assistance in ADL showed that, 2 years later, fewer were living with their parents and even more were married. More of these young adults had children, although at only about half the rate of typical young adults. General health perceptions remained stable, with twice the percentage of respondents reporting fair or poor health compared with typical young adults. Only 17% of respondents reported not having a usual source of care compared with 27% of Kentuckians ages 18-24, but they continued to use the emergency room much more than did typical young adults. Twenty-five percent of survey respondents reported having no health insurance compared with 39% of Kentuckians ages 18-24 and 29% of young adults in the United States. More had completed high school, and 32% had college experience. Two years later, 51% were working compared with 45% initially; 63% of those not working would have liked to have been working, and 53% of those were looking for work. Of great concern is the increase from 23% to 29% of young adults not in school, not working, and not home with children. In general, the study shows that young people with disabilities show progress in markers of adulthood as they move through their 20s just as much as do typical young adults. Nurses can support healthy transitions through assistance in finding adult medical homes, doing preventive screening, teaching knowledge and skills for healthcare planning and management, and planning for continuous insurance. PMID- 17921891 TI - CA-MRSA. The new sports pathogen. AB - Skin infections in athletes caused by community-associated Methicillin-resistant Staphylococcus aureus (CA-MRSA) have been observed within many cities throughout the United States and within many countries throughout the world (Centers for Disease Control and Prevention [CDC], 2003). As the incidence rises in the athletic population, clinicians must learn to identify risk factors for CA-MRSA, diagnosis and treat infections with judicious use of antimicrobial agents and facilitate strategies to limit transmission. Recently, a new consensus guideline for handling CA-MRSA outbreaks in sports has been released by the CDC (Gorwitz et al., 2006). This article includes a review of the evolution of MRSA; distinguishes between healthcare associated Methicillin-resistant Staphylococcus aureus (HA-MRSA) and CA-MRSA; and reviews the diagnosis, management, and prevention strategies to limit transmission of CA-MRSA. PMID- 17921893 TI - Use of wound V.A.C. therapy in pediatric patients with infected spinal wounds: a retrospective review. AB - BACKGROUND: Deep wound infection in patients that have had a posterior spinal fusion for scoliosis is a major complication in pediatric spine surgery. PURPOSE: To explore characteristics of pediatric patients who had a posterior spinal fusion with segmental spinal instrumentation and bone graft and subsequently developed deep spinal wound infections that were treated with wound vacuum assisted closure (V.A.C.) therapy METHOD: Retrospective Chart Review. Characteristics of patients' age, gender, comorbid illnesses, identified bacteria, antibiotics, time on V.A.C. device, infection recurrence, any instrumentation removal, and additional surgical intervention was collected. FINDINGS: There were 249 patients who had spinal fusions from December 2002 through January 2006, and 11 developed an infection after their spinal fusion and had the use of the V.A.C. device. Those who developed infection within 1 year of their fusion had instrument retention. Wound closure occurred in all cases that used V.A.C. therapy. PMID- 17921896 TI - Medicating young or very young patients--Part III. AB - In Part I of this series, the unique considerations about medicating children at different ages were addressed. In Part II, examples of medications used to treat common short-term illnesses in younger patients were examined. In this part, examples of medications used to treat children with two long-term conditions, asthma and attention-deficit/hyperactivity disorder, are identified. Action, contraindications, and safety concerns for these agents are addressed. The challenges and lifestyle changes that these chronic problems pose for children and their parents are also discussed. PMID- 17921898 TI - Deferring planned neck dissection following chemoradiation for stage IV head and neck cancer: the utility of PET-CT. AB - OBJECTIVE: To determine whether combined positron emission tomography and computed tomography (PET-CT) may be of value in deferring planned neck dissections for patients with advanced head and neck squamous cell carcinoma (HNSCC). STUDY DESIGN: Observational study of patients with de novo cervical > or =N2 regional spread of HNSCC in a tertiary care academic medical center. METHODS: Forty-three patients were identified who underwent post-treatment PET-CT within 6 months of completing neoadjuvant chemotherapy combined with radiation therapy (CRT). The PET-CT was "positive" if the radiologist recommended tissue sampling or resection of cervical lymph nodes, or if there was progressive neck disease in the setting of distant metastatic disease. Patients who had positive PET-CT underwent confirmatory biopsy given clinical suspicion for regional cervical metastasis without distant disease. Patients with negative PET-CT were followed clinically and radiographically for a minimum of 5 months (median 18.1 months) after CRT. RESULTS: Ten (22%) of the 43 post-treatment PET-CT studies were positive. Seven of the 10 PET-CT scans (70% of positives) were true-positive given histologically-confirmed residual viable tumor or progressive disease including disease in the neck. The 3 remaining studies (30% of positives) were false-positive PET-CT results, given resolution of fluorodeoxyglucose (FDG) avidity on subsequent imaging or tissue sampling demonstrating absence of viable tumor cells. Of the 33 patients with negative PET-CTs in the neck, 1 patient had absence of FDG-avidity in the setting of malignant disease in the neck (3% false negatives); otherwise, patients with an initially negative PET-CT scan had no recurrences during the study (97% true negatives). This corresponds to a sensitivity of 87.5% (7/8), a specificity of 91% (32/35), a positive predictive value of 70% (7/10), a negative predictive value of 97% (32/33), and accuracy of 91% (39/43) for PET-CT scans in the detection of cervical metastatic disease after CRT. Overall, 37 (86%) of 43 patients were spared neck dissection using this technology without evidence of recurrent disease in the neck at extended follow-up. CONCLUSIONS: Our results suggest that planned neck dissection after CRT for HNSCC may be deferred in favor of serial PET-CT imaging, and that sampling of areas of suspicious FDG-avid uptake can be rationally considered prior to therapeutic neck dissection. These data also suggest that negative PET CT scans are highly reliable for the absence of residual cervical nodal disease. PMID- 17921899 TI - Role of microvascular density in nonlocalizing parathyroid sestamibi scans. AB - OBJECTIVES: Sestamibi scans for localization of abnormal parathyroid glands in patients with hyperparathyroidism are widely used at many institutions. Minimally invasive parathyroid surgery demands accurate preoperative localization imaging; however, nonlocalizing sestamibi scans occur in 15% of patients with primary hyperparathyroidism. It remains unknown why some sestamibi scans fail to localize. We hypothesize that an increase in microvascular density (MVD) within an adenoma will result in rapid tracer washout and a subsequent nonlocalizing scan. This study investigates the role of MVD in sestamibi localization. STUDY DESIGN: Retrospective chart review with immunohistochemical staining and data analysis. METHODS: Medical records of 83 patients who had a sestamibi scan for evaluation of primary hyperparathyroidism and underwent initial parathyroidectomy from 2000 to 2002 were retrospectively reviewed. Patients' age, sex, preoperative imaging results, operative procedure, gland weight, and histologic findings were collected. Immunohistochemistry was performed to assess MVD. RESULTS: Of the 75 preoperative sestamibi scans used, 51 patients had a localizing scan, and 24 were nonlocalizing. Localizing sestamibi scans for primary hyperparathyroidism demonstrated a sensitivity of 94% and specificity of 85%. By identifying multiglandular hyperplasia, nonlocalizing sestamibi scans produced a sensitivity of 83%. The localizing group had a greater percentage of solitary adenomas (94%) compared with the nonlocalizing group (15.6%) (P < .001). The mean gland weight for the nonlocalizing group was less than 398 g compared with the localizing groupweight of 1,113 g (P < .001). The mean MVD for localizing scan group was 229 vessels per high-power field,and the mean for the nonlocalizing scans was 213 vessels per high-power field (P = .2). CONCLUSIONS: MVD does not predict whether sestamibi scans are localizing or nonlocalizing. PMID- 17921900 TI - Role of albumin coating of tympanostomy tubes: long-term clinical evaluation. AB - OBJECTIVE: Our previous work has shown that albumin coating of tympanostomy tubes prevented adhesion of proteins or bacteria on the tube surface in vitro and in a 9-month prospective follow-up study. This study was continued until all tubes were extruded. STUDY DESIGN: A prospective, clinical trial. METHODS: The randomized, double-blind clinical trial had 149 patients. The randomization was revealed after the follow-up period of 9 months. The number of tube sequelae in ears with human serum albumin (HSA)-coated titanium tympanostomy tubes was compared with the contralateral ears with uncoated, otherwise identical titanium tubes. The follow-up continued until all tubes were extruded, followed by evaluation of each tympanostomized patient. RESULTS: No significant difference between the two tube types emerged after the 9-month follow-up. Among the patients younger than 2 years, one of the three typical bacteria causing acute otitis media (AOM), Streptococcus pneumoniae, Haemophilus influenzae, or Moraxella catarrhalis, was found in 45% of all bacterial cultures taken during AOM. However, among patients older than 2, one of these bacteria appeared in 17% of all the bacterial cultures and in 8% of cultures taken during the summer. CONCLUSIONS: After the first 9 follow-up months, no difference was found in the sequelae related to uncoated and HSA-coated tubes. The typical bacteria causing AOM were found less frequently among patients older than 2 years. A profile of tympanostomy patients in Finland will be given. PMID- 17921901 TI - Overall evaluation of effectiveness of type II thyroplasty for adductor spasmodic dysphonia. AB - OBJECTIVE/HYPOTHESIS: To assess the effectiveness of type II thyroplasty with a titanium bridge in adductor spasmodic dysphonia (AdSD). STUDY DESIGN: Retrospective chart review, patient response to a questionnaire on the ease of phonation and voice quality, and pre- and postoperative fiberoptic laryngoscope findings. SUBJECTS: Forty-one patients who underwent type II thyroplasty with a titanium bridge between December 2002 and December 2005 who have been followed for at least 12 months postoperatively. RESULTS: Six patients were male, and 35 were female. The voice was recorded before and at least 6 months after surgery. Initially, 97.6%, 61%, and 48.8% of the patients had a strangulated, interrupted, or tremulous voice, respectively. The mean ratings of strangulation, interruption, and tremor were calculated. The respective mean pre- and postoperative scores were 1.51 and 0.46 for strangulation, 0.76 and 0.05 for interruption, and 0.65 and 0.048 for tremor. In the postoperative questionnaire, 70% of the patients judged their voice as excellent and the remaining patients as improved to good or fair. CONCLUSIONS: Type II thyroplasty is a highly effective therapy for AdSD. The voice in AdSD may roughly be classified into strangulated, tremulous, and interrupted types. The outcome measures justify the continued treatment of AdSD with type II thyroplasty. PMID- 17921902 TI - Scala tympani cochleostomy I: results of a survey. AB - OBJECTIVE: To assess current surgical techniques for scala tympani cochlear implantation among North American surgeons. MATERIAL: A survey was distributed to all cochlear implant surgeons participating in the 2006 William House Cochlear Implant Study Group in Toronto, Canada. Participants were asked to anonymously identify their routine surgical practices. Images of trans-facial recess approaches to the round window and cochlear promontory were used in a multiple choice fashion to assess the surgeon's typical exposure and cochleostomy location. Returned questionnaires were electronically processed and evaluated. RESULTS: Fifty-five (75%) of 73 returned surveys had adequate data validity and availability. Landmark identification and preferred cochleostomy locations varied greatly. About 20% of surgeons selected cochleostomy locations superior to the round window membrane. Cochleostomy size and location appeared to be influenced by surgical experience and whether or not the round window overhang was drilled off. CONCLUSION: This survey clearly documents marked variations in surgical techniques for scala tympani cochlear implantation. Future studies should more clearly define the surgical anatomy of this region for appropriate placement of a scala tympani cochleostomy. These findings may ultimately have an impact on hearing and neural preservation cochlear implant surgeries. PMID- 17921903 TI - Nerve of origin, tumor size, hearing preservation, and facial nerve outcomes in 359 vestibular schwannoma resections at a tertiary care academic center. AB - OBJECTIVE: To determine nerve of origin, tumor size, hearing preservation rates, and facial nerve outcomes in a retrospective cohort study of patients undergoing translabyrinthine (TL), middle cranial fossa (MCF), and retrosigmoid/suboccipital (SO) approaches to vestibular schwannomas (VS). STUDY DESIGN: Retrospective. METHODS: Chart review. RESULTS: Patient charts from 231 TL, 70 MCF, 53 SO, and 5 combined TL/SO procedures for VS were evaluated in 356 patients. The inferior vestibular nerve (IVN) was the nerve of origin in 84 of 359 cases (23.3%), while the superior vestibular nerve (SVN) was the nerve of origin in 36 patients (10%). In 239 of 359 cases (66.6%), the nerve of origin was not identified. Forty patients undergoing hearing preservation surgery had hearing results and nerve of origin data available for review. Functional hearing (<50dB PTA and >50% speech discrimination) was preserved in 10 of 15 patients (75%) with SVN tumors, while only 7 of 25 patients (28%) with IVN tumors retained functional hearing. Facial nerve outcomes and nerve of origin were recorded simultaneously in 109 patients. Seventy-one of 74 patients (95%) patients with IVN tumors achieved a House Brackmann (HB) grade I-III, while 35 of 35 patients (100%) with SVN tumors retained HB I-III facial function. Looking at tumor size versus hearing preservation, functional hearing was preserved in 22 of 49 patients (45%) with <1 cm tumors, and 4 of 20 patients (20%) with 1- to 1.5-cm tumors. For all cases with documented facial nerve function, HB I-III were achieved in 96% of SO, 94% of MCF, and 88% of TL procedures. CONCLUSIONS: Our retrospective data indicated that IVN tumors were twice as common as SVN tumors. The nerve of origin did not affect facial nerve outcomes but did impact hearing preservation rates. Patients with tumors <1 cm in size had the best chance for hearing preservation. Overall facial nerve preservation was excellent with >90% achieving HB 1 to 3 function at final follow-up. PMID- 17921904 TI - Technique, utility, and safety of awake tracheoplasty using combined laser and balloon dilation. AB - OBJECTIVES: Laryngotracheostenosis (LTS) is a condition in which the airway is narrowed between the vocal cords and the carina. We seek to examine whether flexible bronchoscopy with neodymium-doped yttrium aluminum garnet (Nd:YAG) laser incision and balloon dilation tracheoplasty is a practical choice in the management of patients with subglottic or tracheal stenosis. METHODS: A retrospective chart review was performed at a tertiary care hospital. All subjects with laryngotracheostenosis treated between January 1, 2000, and April 2005 who underwent endoscopic Nd:YAG laser incision and balloon dilation tracheoplasty performed using topical anesthesia and intravenous sedation were included. RESULTS: A total of 18 patients comprised the study and 36 procedures were performed without complication. Only one procedure was required by eight subjects, while five subjects required two procedures, three subjects had three procedures, one subject had four procedures, and one subject had five procedures until an adequate stable airway was obtained. The average follow-up was 22 months (range 3-55 months). The average body mass index (BMI) was 32.0 kg/m (range = 20.8-42.2 kg/m) and 11 of the 18 subjects (61.1%) were categorized as obese or morbidly obese by BMI criteria. CONCLUSION: Combined Nd:YAG laser incision and balloon dilation in an awake, spontaneously breathing patient is a safe and effective management tool in the treatment of laryngotracheostenosis. This technique may be particularly beneficial in patients who are at increased risk for general anesthesia such as those with morbid obesity or who have had a history of airway problems during anesthesia. PMID- 17921905 TI - Incision and drainage followed by mattress suture repair of auricular hematoma. AB - OBJECTIVE: Auricular hematoma is a condition requiring early and effective management to prevent pathogenesis of the unsightly cauliflower ear. The objective of this study is to review cases of auricular hematoma and present incision and drainage followed by through-and-through whip-type absorbable mattress sutures without bolsters as an effective treatment. STUDY DESIGN: Retrospective chart review of auricular hematoma cases. METHODS: A 5-year retrospective evaluation of auricular hematomas presenting to an otolaryngology group was performed. Patients' charts were reviewed and data regarding the treatment and follow-up of auricular hematomas were assembled and analyzed. RESULTS: Twenty-two patients were found to present with auricular hematoma. One patient was lost to follow-up. Twenty-eight treatments were performed on 23 ears. Seven hematomas were treated with needle aspiration, two were treated with incision and drainage with iodoform wick placement, and 19 were treated with incision and drainage followed by absorbable mattress sutures. There were five hematoma reaccumulations requiring an additional procedure after treatment by an otolaryngologist. Three followed needle drainage; one followed incision and drainage with wick placement, and one followed incision and drainage with absorbable mattress sutures. CONCLUSION: Incision and drainage followed by through-and-through absorbable mattress sutures appears to be a superior method of treatment with rare reaccumulation of hematoma. This method of treatment was shown to be simple and well tolerated, and it had few complications. PMID- 17921907 TI - Management of the post-tracheostomy scar. AB - Tracheostomy scar management is aimed at filling lost deep tissue bulk, the correction of tracheal skin tug, and a tension-free closure that falls more naturally into the neck folds. Three cases are used to illustrate the major principles involved in the correction of tracheostomy scars. Those reconstructive principles are 1) re-approximation of individual layers of the neck for improved contour and release of tracheal skin tug, 2) filling of tissue deficit, using scar de-epithelialization, muscle flaps, or acellular dermal grafts, 3) excision of hypertrophic scarring or keloids, and 4) horizontal wound closure using simple closure or local skin flaps such as z-plasty. The goals and techniques outlined can resolve skin adherence to the trachea and can improve scar appearance in this noticeable location. PMID- 17921906 TI - Comparison of animal models for head and neck cancer cachexia. AB - OBJECTIVES/HYPOTHESIS: Despite its negative impact on cancer patients, there are few animal models of cancer cachexia. Our hypothesis was that different human cell lines would variably induce cachexia. STUDY DESIGN: Prospective animal study. METHODS: We established two xenograft models in athymic mice and compared these with a cachexigenic cell line, the murine adenocarcinoma 16 (MAC16) cell line. Eight-week-old female, athymic mice were injected with human head and neck cell lines (JHU022, JHU012) and the MAC16 cell line. Body weight, food intake, body composition, leg weights, serum cytokines, and lipid mobilizing factor (LMF) were compared. RESULTS: Mean food intake for all groups was equivalent. Mean percent change in body weight after 18 days was 18%, 19%, 12%, and 3% for control, JHU012, JHU022, and MAC16 experimental groups, respectively. Both JHU022 and MAC16-injected mice showed wasting even when tumor burden was low. In contrast, mice injected with JHU012 developed larger tumors yet lacked evidence of cachexia. These mice demonstrated loss of lean body mass but not fat mass. Serum cytokine levels for interleukin (IL)-1 alpha and IL-1 beta were elevated in JHU022-bearing mice, whereas IL-1 alpha, IL-6, interferon (IFN)-gamma, and tumor necrosis factor alpha (TNF)-alpha were elevated in MAC16-bearing mice. LMF was present in both the JHU022 and JHU012 cell lines. CONCLUSIONS: The JHU022 cell line caused more severe cachexia than the JHU012 cells, suggesting these cell lines may be used to further study cancer cachexia. IL-1 alpha and IL-1 beta in the JHU022 model may be mediators of cachexia, whereas TNF-alpha, IFN-gamma, and IL-6 may be mediators in MAC16-induced cachexia. PMID- 17921908 TI - Probabilities of ossicular discontinuity in chronic suppurative otitis media using pure-tone audiometry. AB - OBJECTIVE: To analyze the likelihood ratios (LRs) and predictive values of preoperative air-bone gap (ABG) levels on the presence of gross ossicular discontinuity (OD) among chronic suppurative otitis media (CSOM) patients. SETTING: Tertiary hospitals. METHODS: Records of 276 patients with CSOM 7 to 75 years old undergoing their first tympanomastoidectomy were reviewed. Association of preoperative audiogram on the presence of OD was analyzed using logistic regression analysis and chi 2 tests. Frequency-specific ABG values were compared with the presence of OD. RESULTS: In CSOM without cholesteatoma, the ABG of 20 dB or less at 500 Hz (LR [-], 0.119; 95% confidence interval [CI], 0.016-0.867) and 30 dB or less at 1 kHz (LR [-], 0.276; 95% CI, 0.087-0.876) decreased probability of OD from 33 to 5.6% and 15.5%, respectively. Air-bone gap levels of greater than 30 dB at 2 kHz (LR [+], 2.8; 95% CI, 1.4-5.9) and greater than 40 dB at 4 kHz (LR [+], 2.2; 95% CI, 1.2-3.9) increased the probability of OD from 33 to 51 to 89%. In the presence of cholesteatoma, the chance of OD was 88%. The ability of ABG to alter probability of OD was not significant in the presence of cholesteatoma. CONCLUSION: Narrow ABG at lower frequencies suggested absence of OD. Wide ABG at higher frequencies suggested presence of OD. Simple tympanoplasty can be done to patients with a small chance of OD as assessed by pure-tone audiometry, whereas a mandatory exploration of the ossicular chain with possible reconstruction was suggested on subjects with a high chance of OD. The presence of cholesteatoma warrants ossicular chain exploration. PMID- 17921909 TI - Three-dimensional fluid-attenuated inversion recovery magnetic resonance imaging investigation of inner ear disturbances in cases of middle ear cholesteatoma with labyrinthine fistula. AB - OBJECTIVE: To investigate the cause of inner ear disturbances in cases of middle ear cholesteatoma with labyrinthine fistula. SETTING: University hospital. STUDY DESIGN: Prospective case study. PATIENTS: Eight patients who were scheduled to undergo surgery for middle ear cholesteatoma with labyrinthine fistula were included in this study. INTERVENTION: Imaging analysis was performed using a 3 dimensional fluid-attenuated inversion recovery (FLAIR) magnetic resonance imaging sequence. MAIN OUTCOME MEASURES: Three-dimensional FLAIR findings were compared with clinical symptoms associated with inner ear disturbance and surgical observations of the fistula. RESULTS: Three-dimensional FLAIR in 6 patients revealed areas of high signal intensity in the inner ears on the affected sides and areas with increased signal after the administration of gadolinium, especially in cases accompanied by acute sensorineural hearing loss. These images were considered to be indicative of breakdown of the blood-labyrinth barrier due to middle ear cholesteatoma. This finding was also present in a patient with no clinical symptoms of inner ear disturbances. CONCLUSION: Three dimensional FLAIR images of the inner ear are valuable in evaluating labyrinthine fistula in patients with cholesteatoma. Future studies are needed to better understand the role of 3-dimensional FLAIR in predicting the severity of inner ear disturbance. PMID- 17921910 TI - Fluctuating corticosteroid-responsive auditory neuropathy/dyssynchrony is suggestive of central nervous system pathology. AB - OBJECTIVE: To highlight diagnostic and management features of auditory neuropathy/auditory dyssynchrony (AN/DS) due to central demyelinating disorder. PATIENTS: A child with AN/DS due to central nervous system pathologic findings. INTERVENTIONS: Audiometry, auditory brainstem response (ABR) test, otoacoustic emission test, magnetic resonance imaging (MRI) with gadolinium contrast, intravenous corticosteroid treatment, antiviral treatment, stereotactic biopsy, and cyclophosphamide immunomodulation. MAIN OUTCOME MEASURES: Pure-tone audiometry, speech discrimination testing, ABR, and MRI. RESULTS: A 12-year-old girl presented with acute sensorineural hearing loss, abnormal ABR, and normal otoacoustic emissions consistent with AN/DS. The hearing loss demonstrated fluctuation and corticosteroid responsiveness. Magnetic resonance imaging and stereotactic biopsy revealed brainstem demyelination consistent with multiple sclerosis. Definitive treatment consisted of cyclophosphamide immunomodulation. CONCLUSION: Although recent focus on pathophysiology of AN/DS has shifted from auditory nerve abnormalities to dyssynchrony within the cochlea, cases associated with fluctuating sensorineural hearing loss and responsiveness to corticosteroid therapy should raise the suspicion of central nervous system pathologic findings. Therefore, it is crucial to obtain brain MRI with contrast enhancement in all patients with AN/DS. This is critical in patients undergoing cochlear implantation because MRI may be contraindicated postoperatively. PMID- 17921911 TI - Spontaneous middle fossa encephalocele and cerebrospinal fluid leakage: diagnosis and management. AB - OBJECTIVE: To evaluate the clinical presentation, operative findings, and surgical management of patients with spontaneous middle fossa encephalocele (SMFE) and cerebrospinal fluid (CSF) leakage repaired using a middle fossa craniotomy (MFC) approach. STUDY DESIGN: Retrospective. SETTING: Tertiary referral center. PATIENTS: Fifteen consecutive patients with 16 SMFE repaired using an MFC approach between January 1999 and April 2006 were included. INTERVENTIONS: Patients were evaluated clinically and radiologically with computed tomography or magnetic resonance imaging. Encephaloceles were approached via MFC, and the cranial base was repaired in multilayered fashion using a variety of materials, including hydroxyapatite cement. Patients were followed clinically after discharge. MAIN OUTCOME MEASURES: Postoperative complications, including CSF leak and the need for surgical revision, are evaluated. Patient factors, diagnostic testing, and operative findings are reviewed. RESULTS: Diagnosis was made using clinical and radiologic evaluation in most patients. Beta2-transferrin testing was occasionally used in the diagnostic workup. Intraoperatively, multiple defects of the floor of the middle fossa were found in more than half of patients. Fifteen SMFE in 14 patients were successfully repaired via MFC alone. One patient required revision with a combined transmastoid/MFC approach due to recurrent CSF leakage. Hydroxyapatite cement was used for repair of the cranial base in 9 patients without complication. CONCLUSION: An MFC approach can be used to repair SMFE with CSF leakage with a high level of success. Hydroxyapatite cement is a safe and useful adjunct to aid in reconstruction of the cranial base defects in cases of SMFE. PMID- 17921913 TI - Client-perpetuated violence and condom failure among female sex workers in southwestern China. AB - OBJECTIVES: This research examined factors associated with condom failure, i.e., slippage or breakage, among female sex workers (FSWs) in China. Special attention was paid to the association between client-perpetuated violence and condom failure. METHODS: Two hundred FSWs were recruited for a community-based voluntary human immunodeficiency virus prevention project. Participants completed a face-to face structured questionnaire that collected information on their sociodemographic characteristics, working conditions, experience of client perpetuated violence, and sexual risk behavior. RESULTS: The prevalence of condom slippage and condom breakage in the 3 months before the survey was reported at 36.2% and 34%, respectively, of all sexual contact in which a condom was used. The prevalence of client-perpetuated violence in the previous year was 68.4%. Logistic regression analysis showed that after adjusting for other factors, condom failure was significantly associated with drug use [adjusted odds ratios (aOR = 4.01)], condom use of coworkers (aOR = 0.39), and client-perpetuated violence [aOR = 2.30 (low violence vs. high violence)]. CONCLUSION: Condom failure is a common problem among FSWs, particularly drug-using sex workers and those who have experienced client-perpetuated violence. On the other hand, condom use of coworkers is negatively associated with condom failure. PMID- 17921912 TI - Does frequency of genital herpes recurrences predict risk of transmission? Further analysis of the valacyclovir transmission study. AB - BACKGROUND: The benefit of suppressive antiviral therapy for reducing the risk of herpes simplex virus (HSV)-2 transmission to sex partners may be enhanced if persons at high risk for transmission can be identified. OBJECTIVE: To determine whether frequency of genital herpes recurrences is associated with increased risk of HSV-2 transmission. METHODS: Analysis of recurrence frequency and shedding frequency (subset) among participants in a randomized controlled trial of valacyclovir 500 mg qd versus placebo for reducing the risk of HSV-2 transmission. RESULTS: Overall, 1484 monogamous HSV-2-serodiscordant couples participated and 41 HSV-2 transmissions occurred during the 8-month trial; 40 were able to provide a history of recurrence frequency. The rate of recurrences per year before study entry did not differ between source partners who transmitted and those who did not, 4.8 versus 5.1, respectively. Similarly, the mean frequency of recurrences observed during the study also did not differ among those who transmitted versus those who did not for placebo recipients (4.4 vs. 4.8) or valacyclovir recipients (1.4 vs. 1.3). Among the 40 source partners who transmitted HSV-2, 8 of 27 placebo recipients and 7 of 13 valacyclovir recipients had no recurrences during the study. CONCLUSION: Clinical assessment of HSV-2 disease severity as defined by the frequency of genital herpes recurrences does not predict the risk of transmission to sexual partners. Though patients with frequent recurrences are most likely to benefit clinically from suppressive therapy, frequency of recurrences is not helpful in identifying persons who are most likely to transmit HSV-2. PMID- 17921914 TI - Non-invasive assessment of coronary artery patency after thrombolysis using serum myoglobin measurements. AB - Timely identification of patients with patent infarct-related artery (IRA) may permit rapid triaging to avoid unnecessary repeat thrombolysis or rescue percutaneous coronary intervention (PCI). Various biomarkers have been proposed to assess IRA patency status. Our group found that the ratio of serum myoglobin levels obtainned before and 60-min after initiation of thrombolytic therapy may provide an early indication of the IRA patency status. A 60-min myoglobin ratio of >or= 4.0 indicated the probability of a patent IRA of 90%, suggesting that follow-up diagnostic modalities such as emergency coronary angiography to determine the IRA status may be unnecessary when these dynamics of myoglobin are observed. PMID- 17921915 TI - Immunological studies in Balkan Endemic Nephropathy. AB - AIM: To re-examine serum complement and immunoglobulin levels in patients with Balkan Endemic Nephropathy (BEN) in the early stage of the disease; to study autoantibodies (anti-thyroid, anti-smooth muscle, anti-mitochondrial, anti parietal, anti-nuclear, and anti-DNA) in these patients; and, finally, to re asses the immunohistology in kidney biopsies from BEN patients. A review of the immunological studies in BEN will be presented. METHODS: Forty-five BEN patients in the early stage of disease, from the South Morava Region, were included in this study. Fifty-five apparently healthy controls, matched for age and sex, from the neighbouring control villages were selected. Serum complement and immunoglobulin levels were determined by the Immunochemistry system of Beckman. Auto-antibodies were detected in the sera by the indirect fluorescence method or by radio-immunoassay (anti-single stranded DNA). Immunofluorescent microscopy was used for kidney samples treated with FITC conjugated antisera for IgG, IgM, IgA, C3, C1q, C4 and fibrin/fibrinogen. RESULTS: Serum C3 was decreased in the sera of patients with BEN compared to healthy controls (p < 0.001), serum IgM was also decreased (p < 0.05). Anti-thyroid antibodies were detected in 5/45 BEN patients, anti-parietal in 7/45 patients and ANA in 2 BEN patients. No anti-smooth muscle, anti-mitochondrial, or anti-DNA antibodies were detected in any BEN patient. Immunofluorescent studies of 10 kidney biopsy specimens showed rare, unspecific, focal, mesangial deposits of C3 and IgM in some glomeruli, and rare deposits, mostly of C3, in tubuli and extraglomerular vessels. CONCLUSION: Humoral immune mechanisms do not appear to play a pathogenetic role in BEN. A few studies on cell-mediated immunity in BEN were performed, and further studies are needed on patients in the early stage of the disease. PMID- 17921917 TI - Non-Hodgkin's lymphomas: immunologic prognostic studies. AB - Non-Hodgkin's lymphomas are malignant lymphoproliferative disorders that originate from B or T-lymphocytes and rarely from NK cells. They represent an extremely heterogeneous group of diseases regarding histologic subtypes, clinical presentations, immunophenotypic profiles, cytogenetic and molecular features and suitable mode of treatment. A clinical indicator of prognosis, the International Prognostic Index, takes into account factors that are mostly linked to patient characteristics (age, performance status) and to disease extension and growth (disease stage, s.lactate dehydrogenase level and extent of extranodal involvement). However, it is clear that differences in clinical features and in treatment responses are a result of the marked genetic, immunophenotypic and molecular heterogeneity that underlie disease aggressiveness and tumour progresssion. We applied IPI (based on pretreatment clinical characteristics) in our group of 136 patients that identified a subgroup of clinical features that remained independently significant in multivariate analyses. In our results IPI turned out to be of prognostic significance for response rate and survival percentages. Based on their number of "poor risk" factors, patients were placed into four IPI risk groups: low (one or no risk factors), low-intermediate (two risk factors), intermediate-high (three risk factors), and high (four or five risk factors) with five years survival rates of 88%; 82%; 18% and 0% respectively. However, one limitation of this prediction strategy is that IPI does not encompass molecular abnormalities of tumour cells, which may play a critical role in determining profoundly different clinical outcomes in patients within the same group as defined by IPI. The aim of this study was to assess the clinical significance and potential prognostic value of the expression of the new immunologic prognostic markers including nuclear proliferating antigen, suppressor and oncogenic proteins, HLA-DR surface antigens, tumour infiltrating T lymphocytes, lymphocyte homing receptor and angiogenic molecules. Immunohistochemistry was used to examine paraffin-embedded tumour tissues for determining the expression of immunologic prognostic markers. Survival analysis showed that serum-high lactate dehydrogenase level, poor performance status (ECOG 3, 4 and 5), high proliferative activity defined as nuclear Ki67 expression greater than 60% of malignant cells and high tumour invasive potential defined by discontinued or loss of Collagen IV were found as strong predictors of poor survival among these patients. These four prognostic parameters determined the New-PI with three risk groups: good (0-1 risk factors), intermediate (2 risk factors) and poor (3 and more risk factors), with predicted five-years survival rates of 88%, 64% and 0%. The New-PI more accurately predicts the outcome than the standard IPI (p < 0,001 vs. p < 0.0001). Based on a single institution series of 136 patients the IPI has proved to be a useful prognostic tool for NHL patients. Addition of new cellular markers into the standard IPI significantly improves risk stratification in NHL. PMID- 17921916 TI - Diagnostic value of biohumoral markers of necrosis and inflammation in patients with right ventricular myocardial infarction. AB - INTRODUCTION/AIM: Patients with right ventricular myocardial infarction (RVMI) and patients with left ventricular myocardial infarction (LVMI) of the anterior wall with ST-elevation (STEMI), due to the profundity and volume of the necrosis, tend to have a more severe and more complicated clinical outcome as well as a higher mortality level compared to patients with myocardial infarction of inferoposterior localization in the left ventricle (IPILK), without the right ventricle being overtaken. C-Reactive protein (CRP) is a sensitive and reliable indicator of acute inflammation and is in good correlation with creatin kinasis (CK) or the enzymes which indicate necrosis markers in acute myocardial infarction (AIM). Because of this, a common biohumoral answer is of greater importance and more reliable both diagnostically and prognostically; it signifies a more severe and more complicated clinical outcome, especially on the rupture of the myocardium. The main goal of this study was to compare the maximum values of enzymes and CRP in patients with RVMI and LVMI who had first STEMI and who were in the acute phase treated with percutaneous transluminal coronary angioplasty (PTCA). METHODS: During a six-year period (2000-05), in the Clinic for Urgent Internal Medicine at the Military Medical Academy, a total of 74 patients included in a prospective study were divided into two groups. The first group consisted of patients with RMI 19 (25.67%), and the second group of patients with LMI 55 (74.33%). The patients in both groups received a percutaneous coronary intervention (PCI), if they had been admitted in the first 4 hrs from the beginning of the chest pain, and if there were no contraindications. All the others received thrombolitic therapy, and a "rescue" PCI if needed, in the next 24-48 hours. The risk factors, clinical outcome, necrosis and inflammation biomarkers (enzymes and CRP), coronary status, restenosis of stent, and intra hospital mortality rate in the first month, as well as a long term prognosis over a period of one year, were analysed. RESULTS: The average age of the patients in the group with RVMI 19 (7 m + 12 f) was 66.1 +/- 11y, and in the group with LVMI 55 (45 m + 10 f) 59.6 +/-13y, with a statistical trend which indicated that the patients with RVMI were older (66.1 +/- 11y vs. 59.6 +/- 13y, p < 0.061) and that women dominated (63.1% vs. 18.8%, p < 0.001). No statistical differences were found between the two groups of patients concerning the length and the appearance of the chest pain before admission to the hospital and the beginning of the PCI treatment, as well as risk factors such as smoking, cholesterol or diabetes. Of the total of 74 patients with the first STEMI as a primary manifestation of a coronary disease, we performed a primary PCI on 58 (78.37%), and a "rescue" PCI on 16 (21.63%) after the thrombolitic therapy during the 24-48h after admission. We had no cases of death either during the primary or the delayed PCI, or in the next 24h following the intervention. During the hospital phase of treatment, in the group with RMI the causes of death were the rupture of the free wall of the right ventricle (1), acute pancreatitis (1), ARDS and hypostatic pneumonia (1), cerebrovascular insult (1). During the following year, one more patient died due to reinfarction of the anterior localization. In the group with LMI, during the hospital phase of treatment 5 (9.09%) patients died: reinfarction (2), rupture of the left ventricle (1), respiratory insufficiency and severe hypostatic pneumonia (1), cerebrovascular insult (1). During the following year, 4 more patients died, sudden death (2), ischemic dilatative cardiomyopathy (2). The total mortality rate over a one-year period of observation in the group with LMI was 9 (16.3%), and in the group with RMI 5 (26.3%). Radionuclide ventriculography (RNV) was performed in the acute phase of myocardial infarction from 7-14 days after PCI and after 6 months in both groups as an independent indicator of the ejection fraction (EF) of both ventricles. The given results show that a statistically proven significant difference exists in the recovery of the right ventricle in acute phase RMI (49.1 +/- 7.9 vs. 35.4 +/- 10, p< 0.001), as well as after 6 months (49.2 +/- 9.7 vs. 38.3 +/- 11.2, p < 0.010) in patients with RMI. CONCLUSION: Primary PCI should be done whenever it is possible with all patients who have a great volume and depth of necrosis, especially if that is the first manifestation of a coronary disease and the first acute STEMI, as were all of our patients in both groups. Our results show that older patients with RMI, and dominantly women, have a more severe and more complicated clinical outcome in the acute phase of RMI compared to patients with LMI of the anterior wall. In the longer prognosis of this case, they have a quicker and a more complete recovery of the right ventricle due to which they have a better immediate and long term prognosis, but demand careful overseeing and energetic treatment in the acute phase of the myocardial infarction, especially considering that their treatment is often specific compared to patients with an infarction of the left ventricle. PMID- 17921918 TI - Mycophenolate mofetil in the treatment of glomerular diseases. AB - AIM: Treatment of primary glomerular diseases may be unsuccessful or have potential toxicities. Mycophenolate mofetil (MMF) is a new, relatively non-toxic drug. It has been introduced as an immunosuppressive drug, but it also has effects on non-immune cells (vascular smooth muscle cells, fibrocytes). Therefore, we evaluated the use of MMF for the treatment of glomerular diseases at different stages of the disease. METHODS: The daily dosage of MMF was 2 for the first 6 months and 1.5 g for a further 18 months, combined with steroids. The follow-up period was two years. RESULTS: 18 patients with lupus nephritis were treated. Patients with a high histological activity index showed a significant decrease of serum creatinine (p < 0.05) and proteinuria (p < 0.01), while patients with a high chronicity index showed only a decrease of proteinuria (p < 0.05). 15 patients with membranous nephropathy were treated. They showed stable renal function and a significant decrease of proteinuria (p < 0.05). Complete remission was achieved only in patients with MMF as a first choice drug. 4 patients with focal segmental glomerulosclerosis did not show any significant decrease of proteinuria, while the nephrotic syndrome in minimal change nephropathy (3 patients) showed a complete recovery. Partial improvement of the nephrotic syndrome was noted in 5 patients with membranoproliferative glomerulonephritis and in 4 patients with crescentic glomerulonephritis. Patients with crescentic glomerulonephritis also presented a significant decrease of serum creatinine (p < 0,05). MMF in 3 patients with IgA nephropathy grade I showed a significant improvement of the nephrotic syndrome. In grade III (5 patients) the response was partial. CONCLUSIONS: We can conclude that MMF in our patients showed both actions, as an immunosuppressive drug in the early stages of the disease, and as an anti-fibrotic agent in the chronic phase of the disease. PMID- 17921919 TI - Progression of diabetic nephropathy: value of intrarenal resistive index (RI). AB - We used duplex Doppler analysis to determine whether the intrarenal RI can be used as a predictor in patients with diabetic nephropathy. Intrarenal resistive index (RI) values were obtained from intraparenchimal arteries of both kidneys, either the arcuate or interlobar arteries. Clinical parameters and renal function were also evaluated at baseline and after three and six months. Forty patients with diabetic nephropathy were divided into two groups based on their intrarenal RI values: group 1 had values of >or=70 and group 2 had values <70. Progression of renal function (delta creatinine clearance, delta CCr) was estimated by linear regression of the slope of decline of CCr plotted against time. At baseline, sixteen patients (40%) had an intrarenal RI value >or=70. Eight patients (50%) of them had a decline in renal function after six months. In comparison, among patients with intrarenal RI values <70 (n=24), only 2 had a decline in renal function. In multivariate regression analysis, proteinuria, lower baseline CCr and RI were independent predictors of declining renal function. An intrarenal RI value of >or=70 identifies diabetic patients at risk of progressive renal disease. The RI of interlobar arteries seems to be a dependable marker of intrarenal changes and can be used as a non-invasive, easily available parameter of the evolution in patients with advanced clinical diabetic nephropathy. PMID- 17921921 TI - Relationship between myocardial viability and improvement in left ventricular function and heart failure symptoms after coronary artery bypass surgery. AB - BACKGROUND: The evaluation of myocardial viability is an important preoperative parameter, predictive of improvement in regional and global left ventricular (LV) function after coronary artery bypass surgery (CABG). However, whether the presence of viability is also associated with relief of heart failure symptoms after revascularization is not always certain. The aims of the study were to define the relationship between extent of viable myocardium and improvement in LV function after CABG and to determine whether preoperative viability testing can predict improvement in heart failure symptoms. METHODS: Eighty-five consecutive patients with ischemic cardiomyopathy (mean LVEF 35%) undergoing surgical revascularization were studied with a Tc-99m sestamibi one-day rest/nitrate enhanced myocardial perfusion SPECT imaging (MPI) to assess viability. Regional and global function were measured before and 16 -/+ 6 months after revascularization. We have used the Bull's eye quantitative analysis of MPI scans and 17 segment model of LV function and perfusion evaluation. Heart failure symptoms were graded according to the New York Heart Association (NYHA) criteria, before and 16 -/+ 6 months after revascularization. RESULTS: The number of viable segments per patient was directly related to the improvement in LVEF after revascularization (r 0.79, P < 0.01). Patients with > 4 viable segments representing 24% of the left ventricle yielded the sensitivity of 83% and specificity of 79% respectively for predicting improvement in LVEF. Furthermore, the presence of four or more viable segments predicted improvement in heart failure symptoms after revascularization, with positive and negative predictive values of 79% and 74%, respectively. CONCLUSION: The presence of substantial viability (four or more viable segments, 24% of the left ventricle) on myocardial perfusion gated SPECT imaging in patients with ischemic heart failure before CABG surgery has significant correlation with the improvement in LVEF and heart failure symptoms postoperatively. PMID- 17921920 TI - Association between haemoglobin level and all-cause mortality in haemodialysis patients: the link with inflammation and malnutrition. AB - Although anaemia management has improved in haemodialysis (HD) patients in recent years, many of them still have haemoglobin (Hb) levels below the current recommendations. The consequent anaemia could be one of the links between malnutrition and inflammation, and higher mortality in HD patients. The study objective was to determine the relationship between Hb levels and outcome in patients undergoing HD, accounting for inflammation and malnutrition. We retrospectively analysed a total of 236 patients on HD between January 2003 and December 2005, classified by absence or presence of inflammation and malnutrition (defined as serum albumin levels < 40 g/L and CRP > 8mg/l). Serum levels of Hb, ferritin, creatinine, cholesterol, triglycerides, HDL (high-density lipoprotein cholesterol), LDL (low-density lipoprotein cholesterol), albumin and CRP were measured monthly, fibrinogen was measured every third month. Over the period of three years, 73 out of 236 patients (30%) had died, most from cardiovascular diseases (62%). Presence of inflammation and malnutrition (in 44% of patients) was associated with older age (60.69 -/+ 12.46 vs. 54.52 -/+ 12.37, p = 0.0002), lower levels of Hb (99.53 -/+ 14.97 vs. 111.86 -/+ 10.38 g/l, p = 0.0000), creatinine (835.88 -/+ 179.84 vs. 1069.98 -/+ 821.23-/+mol/l, p = 0.0047), albumin (36.58 -/+ 3.41 vs.40.32 -/+ 2.82 g/l, p = 0.0000), cholesterol (4.32 -/+ 1.04 vs. 4.75 -/+ 1.09 mmol/l, p = 0.0025) and higher levels of fibrinogen (4.94 /+ 1.18 vs. 4.29-/+0.91g/l, p=0.0000) and CRP (30.42-/+29.47 vs. 5.24-/+4.89 mg/l, p=0.0000). The Kaplan-Meier analysis showed that, irrespective of the absence or presence of inflammation and malnutrition, the all-cause mortality was higher in patients with Hb <110g/l (Log-Rank, p=0.00147; p=0.00222). On the other hand, Kaplan-Meier showed that, irrespective of the absence or presence of anaemia (Hb > 110g/l and Hb < 110g/l), the all-cause mortality was higher in patients with the presence of inflammation and malnutrition (Log-Rank, p=0.00222; p=0.00263). The Cox proportional hazard analysis, adjusting for age, showed that only lower serum levels of Hb and higher CRP were associated with all-cause mortality (chi-square=110,306, p=0.0000). Our findings confirm the association of Hb levels < 110g/L with higher mortality among maintenance HD patients, especially in patients with the presence of inflammation and malnutrition. Further investigation of the relationships among anaemia, inflammation and malnutrition and survival is warranted. PMID- 17921922 TI - Massive deep lobe parotid neoplasms and parapharyngeal space-occupying lesions: contemporary diagnostics and surgical approaches. AB - Regional anatomical peculiarity and the evident increasing incidence of parapharyngeal space neoplasms, have established the necessity of an adequate diagnostic approach for their verification and competent preoperative assessment. Tumours of the parapharyngeal space encompass a wide variety of benign or malignant neoplasms of different origins. The clinical course of 26 patients with parapharyngeal space neoplasms was analysed. A standard diagnostic algorithm for parotid neoplasms was performed, followed by adequate surgical procedure with modifications. Arteriography was used only when CT or MRI suggested a glomous tumor, or possible involvement of the carotid artery, or when those tumours put the carotid artery at risk of surgical injury. The results found from statistical evaluation revealed a higher significance of CT and MRI for tumor extension, localization and comprehension. MRI was dominant for determining tumor character and facial nerve involvement. A transparotideal approach including two modifications was confirmed as being the most effective. When a parapharyngeal tumor is detected deep lobe parotid neoplasms are frequently to be expected. Exact preoperative planning with imaging techniques and a posterolateral parotidectomy approach with an angular mandibular osteotomy produces much better postoperative results in the surgery of deep lobe parotid tumours. PMID- 17921923 TI - Insulin pump therapy with continuous glucose monitoring improves metabolic control in brittle type 1 diabetes. AB - AIM: to evaluate the combination of insulin pump therapy and continuous glucose monitoring in outcome on metabolic control in patients with brittle type 1 diabetes. MATERIALS AND METHODS: Insulin pump therapy was initiated in eleven brittle type 1 diabetics with poor metabolic control (mean Hba1c = 9.6%). Metabolic control was evaluated with CGMS and HbA1c in the following 6 months. RESULTS: Glycated haemoglobin showed a reduction in 1.4% in the 6 months following initialisation of pump therapy. Physical activity, various foods and insulin were tested with CGMS. There were no severe hypoglycaemia and occasional postprandial hyperglycaemia, where patients and their family learned the practical issues of carbohydrate counting. During the next 6 months on pump therapy, the patients successfully managed their diabetes. CONCLUSIONS: Insulin pump therapy can be initiated and used effectively in brittle type 1 diabetics to improve metabolic control and quality-of-life. When diabetes and pump management are appropriately individualized, this kind of therapy can help type 1 diabetics to achieve and to sustain metabolic control. Lifestyle flexibility, quality-of life improvement, and independence can be maintained throughout young adulthood. PMID- 17921924 TI - Dominant species of the gastropod fauna from the littoral region in Lake Ohrid of R. Macedonia. AB - The class of Gastropoda is one of the Lake Ohrid' macrozoobenthic groups, which characterize highest level of endemism. Since the first published references until now, studying the Lake Ohrid's snail fauna represents huge scientific challenge for many malacologists from the world. The high percent of endemic as well as relic forms among the lake's gastropods could be explained by the processes of intralacustric speciation during the history and evolution of this aquatic ecosystem. These processes no doubt have been enabled by the complexity and stability of the Lake's basin. This work represents the results of the dominant species, from both qualitative and quantitative sense of the gastropod fauna from 19 investigated littoral localities of Lake Ohrid. The results from the investigations on gastropod fauna has shown that following species: Chilopyrgula sturanyi, Radix relicta and Valvata stenotrema quailtatively predominates in the samples from the littoral region of the Macedonian part of Lake Ohrid. The quantitative analyses (according to their presence on m2), has shown that the following species predominate: Chilopyrgula sturanyi (6879 No x m2), Theodoxus fluviatilis dalmaticus (6412 No x m2), Pyrgohydrobia grochmalickii (5504 No x m2) and Valvata stenotrema (5009 No x m2). PMID- 17921925 TI - Oxidative stress in patients with beta-thalassemia major. AB - The aim of this work is to study the level of oxidative stress in blood of beta thalassemia major patients with transfusional iron overload and chelation therapy as a central pathological process. Beta-thalassemia major results in an increase in the concentration of lipid peroxidation products in blood plasma of more than 100% and in the intensity of chemiluminescence - about 20% in comparison to healthy controls. The activity of the antioxidant enzyme superoxide dismutase in the blood of beta-thalassemia major patients is decreased by more than 30% and the total antioxidant activity is diminished by about 70% compared to controls. Experimental data confirm the progression of oxidative stress in patients with beta-thalassemia major: activation of free radical processes and lipid peroxidation, decreased antioxidant capacity. Strong oxidative damage and essential alternations define these parameters as sensitive markers of oxidative stress in patients with beta-thalassemia major. The combination of effective iron chelatory agents with natural or synthetic antioxidants can be extremely helpful in clinical practice in the regulation of the antioxidant status of patients with beta-thalassemia major. PMID- 17921926 TI - Radiological and "imaging" methods in TNM classification of non-small-cell lung cancer. AB - Lung cancer is the most common worldwide malignant disease according to its incidence and mortality. The aim of our study was to evaluate the diagnostic value of the radiological and imaging methods, according to the TNM classification, compared to postoperative histological diagnosis. Thirty-seven patients with pulmonary carcinoma were studied prospectively using native chest radiography (PA and LL view), computed tomography (CT) and magnetic resonance imaging (MRI) during ten days before thoracotomy. Radiological and imaging findings were reviewed separately and results were compared with surgical and pathohistological findings on the basis of the TNM classification. All patients underwent chest x-rays, CT was performed in 36 patients and MRI in 12 of them. Imaging methods (CT and MRI) showed more accuracy in sensitivity and specificity compared with the native chest radiography in a great percentage. Generally no statistically significant differences were found between the two imaging methods for the evaluation of tumour extent (T) or lymph node metastases (N). MRI was slightly superior to CT in determination of the chest wall extent of the tumour. In conclusion CT remains the imaging modality of choice both for assessing patients with abnormal chest radiographs suspected of having lung cancer, and in staging patients with histologically proven pulmonary carcinoma. PMID- 17921927 TI - Electrophysiology in neuropsychiatric research: a network perspective. AB - A growing number of anatomic and physiologic studies have shown that parallel sensory and motor information processing occurs in multiple cortical areas. These findings challenge the traditional model of brain processing, which states that the brain is a collection of physically discrete processing modules that pass information to each other by neuronal impulses in a stepwise manner. New concepts based on neural network models suggest that the brain is a dynamically shifting collection of interpenetrating, distributed, and transient neural networks. Neither of these models is necessarily mutually exclusive, but each gives different perspectives on the brain that might be complementary. Each model has its own research methodology, with functional magnetic resonance imaging supporting notions of modular processing, and electrophysiology (eg, electroencephalography) emphasizing the network model. These two technologies might be combined fruitfully in the near future to provide us with a better understanding of the brain. However, this common enterprise can succeed only when the inherent limitations and advantages of both models and technologies are known. After a general introduction about electrophysiology as a research tool and its relation to the network model, several practical examples are given on the generation of pathophysiologic models and disease classification, intermediate phenotyping for genetic investigations, and pharmacodynamic modeling. Finally, proposals are made about how to integrate electrophysiology and neuroimaging methods. PMID- 17921928 TI - Predictors of therapeutic response to treatments for depression: a review of electrophysiologic and dichotic listening studies. AB - There are few clinical or biologic predictors of response to treatments for depression. This article reviews growing evidence that electrophysiologic and neurocognitive measures of brain function may be of value as predictors of therapeutic response to antidepressants. Initial studies using dichotic listening, quantitative electroencephalography, or event-related brain potential measures have found differences between treatment responsive and nonresponsive subgroups of depressed patients. The neurophysiologic basis for these differences and the potential clinical utility of electrophysiologic and dichotic predictors of treatment outcome remain to be determined in future studies. PMID- 17921929 TI - Magnetoencephalography and magnetic source imaging: technology overview and applications in psychiatric neuroimaging. AB - Magnetoencephalography (MEG) is an electrophysiologic brain imaging technology that has been applied to the study of mental illness, particularly schizophrenia. Like electroencephalography, it provides excellent temporal resolution, and in combination with magnetic resonance imaging, can also provide good spatial resolution. Studies of the auditory system in schizophrenia using MEG have demonstrated an abnormality in functional cerebral asymmetry, in which persons with schizophrenia typically show reduced, or reversed, cerebral asymmetry compared with normal subjects. This abnormality is sex-specific; it is more pronounced in males with schizophrenia. These findings have not been demonstrated using other neuroimaging strategies. Thus, MEG appears to offer a unique and valuable contribution to psychiatric neuroimaging. Current research and clinical applications of MEG are limited, however, by the high cost of instrumentation. The cost of MEG systems should improve as more applications are developed, in schizophrenia as well as other neuropsychiatric conditions, and hospitals begin to invest in the technology. PMID- 17921930 TI - Multimodal imaging in psychiatry: the electroencephalogram as a complement to other modalities. AB - The use of different imaging modalities provides the clinician and researcher with different views of anatomy and physiology at unprecedented levels of detail. Multimodal imaging allows for noninvasive measurement of structure and function in humans during complex behavior, and thus provides information about the inner workings of the brain previously unavailable. The present paper examines the various imaging techniques available, and describes their application to the clinic-in the case of epilepsy-and to research-in the case of schizophrenia. Because the electroencephalogram has a dynamic response in milliseconds, it provides the best temporal sensitivity of functional measures of brain activity. When coupled with high-resolution magnetic resonance imaging measures of brain structure, this multimodal approach provides a powerful tool for understanding brain activity. Clinically, the use of multimodal imaging has provided greater precision in localization of the epileptogenic focus. For researchers attempting to determine the underlying causes of schizophrenia, the use of multimodal imaging has helped lead the field away from a specific lesion view to a more distributed system abnormality view of this disorder. PMID- 17921931 TI - What would it take for electrophysiology to become clinically useful? AB - In this article we discuss the procedures that should be followed in order to develop diagnostic tests based on electrophysiologic parameters that would be useful to practicing psychiatrists in their efforts to diagnose mental disorders. To do this, we start by giving an overview of receiver operating characteristic (ROC) analysis, with which a candidate diagnostic test can be characterized mathematically. This methodology allows for an estimate of the overall discriminating power of the diagnostic test and thus can be used to compare the performance of one diagnostic test with that of another. However, ROC analysis cannot be used for optimizing a given diagnostic test (ie, determining the "best" cut-off) and, because it does not incorporate Bayes' theorem, it cannot be used to compare the performances of diagnostic tests when it is given to different risk groups for the target disorder. Both of these problems can be solved by using the concept of information theory. The equation for information gain automatically takes Bayes' theorem into consideration and also provides an intrinsic criterion for finding the cut-off that best discriminates between individuals who do and those who do not have the target disorder. Neither ROC analysis nor information theory are model dependent; however, if the equation for the frequency distribution of the electrophysiologic variable is known for the disease-positive and disease-negative populations, the calculations are greatly simplified. Therefore, the assumption is often made that both of these distributions (or transformations of the distributions) are Gaussian. However, studies have shown that the results of ROC analysis are quite robust to deviations from normality of the underlying distributions. PMID- 17921932 TI - Endoscopic ultrasound in gastroenterology and beyond. New challenges in the XXI century. PMID- 17921933 TI - [Endoscopic ultasound: history and techniques]. AB - Endosonography is an imaging technique based on the unification of ultrasonography and edoscopy. EUS became part of clinical practice at the beginning of the 80s due to its superior capacity to study the pancreas with respect to abdominal ultrasound. In recent years, there is a growing interest in EUS as shown by the number of scientific articles and changing clinical practice regarding diagnostic and therapeutic algorithms (bile duct stones, cancer staging) and palliation. PMID- 17921934 TI - Endoscopic ultrasound teaching and learning. AB - Scientific publications in the field of endoscopic ultrasound (EUS) are raising up since the last twenty years. The value of EUS is directly proportional to the training, skill and experience of the endosonographer. In a recent series, the lowest annual number of examinations requested per year might be around 200-250 procedures. For achieving competence in all the fields of EUS, they recommended a minimum of 150 supervised cases, 75 of them being devoted to pancreaticobiliary diseases and 50 to fine needle aspiration (FNA). Many studies have yet confirmed the importance of the learning curve to improve the EUS accuracy. The accuracy of EUS staging of esophageal cancer was significantly improved over an 8-year period for T staging from 64% to 90% but not for N staging. Considering pancreaticobiliary diseases, some series reported the influence of training on accuracy of EUS-guided FNA of pancreatic masses. Even if the exact type and modalities for EUS training remained to be determined, the usefulness of a period of training in a center performing more than 200 procedures per year is confirmed. Improving cognitive EUS learning and technique required theoretical teaching and practical teaching with observation during live demonstration and echoendoscope hands-on in humans with an expert supervision or with different simulator models. In France, formal EUS training has been underway since 1991, with a two-year intensive program including 4 weeks of theoretical training and 20 one-day practical sessions per year. This program was updated in 2004 to include 3 weeks of theoretical course work including numerous video sessions and 1 week of practical sessions. Theoretical courses are given primarily by gastroenterologists and EUS experts, but also by pathologists, surgeons, anatomists and radiologists. Practical sessions include 4 days of live demonstrations and computed-based simulator (EUS mentor; Simbionix, Tel Hashomer, Israel) and one day with a swine model. Simulators, either computed-based simulator, Erlangen model, or live pigs are believed to shorten the learning phase of EUS. The live pig model was chosen for EUS credentialing because it appears to be similar in many respects to the human anatomy, especially for the pancreas, portal and mesenteric vessels, celiac axis, left kidney and spleen. In addition, for trainees, it often appears more exciting and stimulating than virtual model. However, EUS training on a swine model is recognized to be the more expansive way for EUS teaching. Between the pre and post-test, EUS fellows significantly improved their competence for the visualization of anatomical structures i.e. vena cava, mesenteric/splenic vein, celiac axis, pancreas and bile duct. At the end of the day-session, trainees were mostly able to recognize and follow these anatomical structures with both endoscopes. Interventional EUS learning was also assessed. A significant decrease in procedural duration and a significant increase in FNA precision were demonstrated for the puncture of a hilar liver lymph node. PMID- 17921935 TI - Instruments and accessories for diagnostic endoscopic ultrasound (radial scanning and miniprobes). AB - Endoscopic ultrasound (EUS) is a relatively new technology developed in the early 80's, but introduced into the clinical practice only in the late 80's. Nowadays there is an increase in demand for EUS examinations and services, because this technique is more and more recognized as an important and integral part in the diagnostic and staging algorithm of many gastrointestinal (GI) and nongastrointestinal pathologies. Even if today the EUS panorama enrich itself with a large number of different instruments, mechanical and electronic scopes, radial and linear ones, several types of miniprobes and specialty probes, for diagnostic purposes the dedicated radial scanning instruments remain the more adopted choice while miniprobes can have applications only in niche areas, like superficial GI cancers staging, small subepithelial lesions evaluation or intraductal ultrasound and some technological advances are needed to solve the potential drawbacks of this miniaturized probes. At last the EUS equipment borrowed most of the electronic sophistication from the mainstream ultrasonography, this made the choice of an EUS instrument even more difficult and challenging, but you must consider that there is no right or wrong equipment, there is only the right instrument for that specific clinical need or situation. PMID- 17921937 TI - [The nurse's role in endoscopic ultrasonography]. AB - The role of trained nurses in endoscopic ultrasonography (EUS) examination is very relevant for the quality and results of EUS. Nurses have many tasks in EUS. To prepare the endoscopic room with the right instrument and necessary devices for examination of the upper or lower GI tract is very important. It is also crucial that the nurse gives the right information about the procedure to the patient, to relieve anxiety and to give explanations about the modality of the endoscopic procedure. During the procedure the nurse must help the endoscopist and, when indicated, the anesthesist. After the completion of the procedure, the nurse must carry-on with the reprocessing of the endoscopic instrument and of the devices. Specialized and dedicated nurses who follow courses to keep up-dated are indispensable in this field because of the constant evolution of the endoscopic instruments and techniques. Possible complications must be kept in mind to be recognized and to be treated in an early phase. The endoscopy-trained nurse must know the anatomy of the examined segments but should however integrate this knowledge with the care of the patients. Nurses should also contribute to clinical research regarding EUS. PMID- 17921936 TI - [Instruments and accessories for interventional endoscopic ultrasonography]. AB - Endoscopic ultrasound-guided (EUS) biopsy is now an established method for acquisition of cytohistological specimens. Some other procedures can be performed successfully in most instances (e.g., celiac plexus block, pseudocystic drainage), and some are being performed but need further refinement or validation (e.g., pancreatic and biliary drainage, ablation of pancreatic cyst neoplasm). The present review describes the interventional endoscopes, the needle systems and the specific devices for other interventional procedures available on the market. PMID- 17921938 TI - [Management and preparation of endoscopic suite, patients, relatives. Instrumentarium reprocessing]. AB - Endoscopic ultrasonography (EUS) investigates the inner side of the digestive tract and the adjacent structures, associating the endoscopic image to the ultrasonographic vision made by a miniaturized ultrasonograph. The technological innovations and a greater attention to the users, have made more complex the organization, the process and the management of the patients. In such panorama, the technical operator of endoscopy, is the competent professional that coordinates the whole necessary organization for diagnostic-therapeutic interventions assuring their feasibility, guaranteeing efficiency and safety of environmental hygiene and strumentario and a specific and competent relief approach to the patients and their relatives. PMID- 17921939 TI - [Risk management: relationship with the patient's relatives, who does what?]. AB - Risk management is the systematic process of identification, evaluation and treatment of current and potential risks. In the last decades it has acquired a significant importance also in the medical field, where it aims to increase patient' safety, to reduce adverse events and costs, thus improving patients' outcomes. Gastrointestinal endoscopy is a clinical field where operative procedures are carried out; some of them may have complications, which can lead to legal cases; among such techniques endoscopic ultrasonography is acceptably safe. The aim of the article is to highlight some aspects of the risk management which are related with the patient's relatives, in the field of endoscopic ultrasonography: from the gathering of the case history to the required information for a safe discharge. Communication skills and attention given to the information process, can allow the reduction of clinical risks for patients undergoing endoscopic procedures. PMID- 17921940 TI - New frontiers in therapeutic EUS: dreams, style exercises or actual possibilities. AB - The term therapeutic endoscopic ultrasound (EUS) refers to direct ultrasound guided placement of pharmaceuticals, devices and biological agents for the purpose of treating a disease or symptoms. The vast majority of EUS procedures in the world are performed for diagnostic purposes, although these lead directly to therapeutic decisions such as whether to treat diseases by local (including endoscopic) resection, surgery or chemotherapy. EUS imaging is providing directions for therapies in different conditions, such as 1) gastrointestinal (GI) diseases including luminal GI malignancies, 2) management of pancreatic pain by performing the celiac plexus block, 3) drainage of obstructed biliary and pancreatic ducts when standard ERCP methods have failed, and 4) lesions surrounding the gut wall, including adjacent organs (pancreas liver, etc) and mediastinum (e.g. non-small cell lung cancer staging). Experimental promising EUS applications include delivery of cytotoxic agents such as chemotherapy, gene therapy, and radioactive seeds. This article will review the diagnostic and therapeutic role of EUS in gastrointestinal as well as extra-intestinal diseases. PMID- 17921941 TI - Esophageal cancer: biology, natural history, staging and therapeutic options. AB - The distribution of adenocarcinomas and squamous cell carcinomas in esophageal cancer (EC) has changed, and focus directed towards tumors of the distal esophagus and the esophagogastric junction. The genetic events leading to EC are not fully clarified, but important risk factors have been identified. Accurate pretherapeutic tumor evaluation should be provided for a multi-disciplinary and individually tailored patient management programme. Endoscopic ultrasonography (EUS) and PET-CT play an important role in the assessment of disease, evaluation of treatment response and during follow-up. PMID- 17921942 TI - Radial EUS: the clinical impact of T staging. AB - Endoscopic ultrasound staging of gastrointestinal and pancreaticobiliary cancers is important in guiding the choice of an appropriate treatment strategy such as endoscopic mucosal resection, surgery or palliative chemotherapy. This review will summarize the principles of endoscopic ultrasound T staging using a radial echoendoscope, elaborate on the accuracy rate in T staging, and discuss the clinical impact of endoscopic ultrasound T staging in the context of esophageal, gastric and pancreaticobiliary cancers. PMID- 17921943 TI - Linear EUS: the clinical impact of N staging in esophageal carcinoma. AB - This review article focuses on nodal staging of esophageal cancer. Transesophageal endoscopic ultrasound (EUS) is the most accurate technique for preoperative local-regional staging of esophageal carcinoma (TN staging), once the CT and/or the PET scan have excluded the presence of distant metastasis. EUS guided fine needle aspiration (EUS-FNA) helps improve diagnostic accuracy in esophageal cancer lymph node staging. In certain subgroup of patients who present with a number of EUS lymph node criteria, EUS FNA may be avoided without affecting diagnostic accuracy. In tumors of the distal esophagus detection and biopsy of celiac lymph nodes may be successfully performed by EUS and EUS FNA, and important therapeutic decissions may be derived from such a practice. PMID- 17921944 TI - Lung cancer: biology, natural history, staging and therapeutic options. PMID- 17921945 TI - Endoscopic ultrasound and staging of non-small cell lung cancer. AB - Accurate cancer staging allows the most favorable therapy and prognosis of a neoplastic process, and allows consistency when performing clinical trials for different stages of tumors. In the case of non small-cell lung cancer (NSCLC), the most important parameters for optimal treatment and prognosis are the presence of cancer spread to the lymph nodes within the mediastinum, and to distant organs. Endoscopic ultrasound (EUS) has become a significant tool for the assessment of mediastinal lymph nodes, and in some cases, distant organ metastases because of its minimally invasive access to these sites through a trans-esophageal or trans-gastric approach. The capability of performing fine needle aspiration (FNA) has greatly improved the accuracy and popularity of EUS for lung cancer staging. This review will outline the basic principals of lung cancer staging, EUS-FNA techniques, and role of EUS-FNA in lung cancer staging. PMID- 17921947 TI - [Endoscopic ultrasound-fine needle aspiration (EUS-FNA) for pancreatic lesions: effectiveness in clinical practice]. AB - Endoscopic ultrasound-fine needle aspiration (EUS-FNA) was shown to be a highly reliable and a very effective diagnostic technique, both based on data from clinical trials and from large clinical practice studies. EUS-FNA results are reported to be in good-to-very good agreement with the final diagnosis, and the agreement significantly exceeded the chance agreement. The overall sensitivity and specificity of EUS and of EUS-FNA are very good. EUS-FNA is an effective diagnostic technique for the evaluation of pancreatic lesions, either reported with other imaging tests or suspected on the basis of clinical and biochemical features. EUS-FNA may be performed in most cases, and the results of EUS-FNA are particularly important for their excellent positive predictive value. Nonetheless, in a few cases EUS-FNA can not be feasible, or can give false negative or inconclusive RESULTS: The main practical consequence is that before referring patients to surgeons or oncologists, EUS-FNA should be considered as the best diagnostic strategy, since tissue is still the issue' . In a prospective two-centers consecutive series from Italy, FNA did not give any false positive diagnoses of malignancy, and reduced the number of indeterminate diagnoses; moreover, FNA significantly increased the specificity of diagnosis, while sensitivity was unchanged. PMID- 17921946 TI - The complete ''medical'' mediastinoscopy (EUS-FNA + EBUS-TBNA). AB - Diagnosis of indeterminate mediastinal masses and staging of lung cancer poses a significant challenge. Options for tissue diagnoses include computed tomography (CT)-guided percutaneous biopsy, transbronchial fine-needle aspiration, mediastinoscopy/mediastinotomy or thoracoscopy, but these investigations have limitations in terms of tissue yield, safety profile and cost. Trans-esophageal endoscopic ultrasound scanning (EUS) is a new minimal invasive method that provides high resolution imaging of the mediastinum using high frequency ultrasound probes attached to the tip of a flexible endoscope and offers in addition the facility of fine needle aspiration (EUS-FNA) or tru-cut biopsy (TCB) under real-time ultrasound guidance. EUS-FNA allows access to the posterior mediastinum and tissue acquisition under real-time ultrasound guidance through the oesophageal wall. Indications of EUS-FNA in the mediastinum is to obtain a diagnosis from an unknown primary lesion or to sample tissue from mediastinal lymph nodes in order to stage lung cancer or to diagnose other diseases involving lymph nodes of the mediastinum eg. TB, Sarcoidosis, histoplasmosis or metastases from a vide range of cancers. If lymphoma is suspected EUS-TCB of an enlarged mediastinal lymph node is preferred. EUS- FNA is safe, can be done on an outpatient basis, is well tolerated and provides an excellent diagnostic yield with a sensitivity of more than 90% and a specificity of 100%. Compared to CT, PET, mediastinoscopy as well as transbronchial aspiration, EUS-FNA is found to be significant more accurate for staging of non-small cell lung cancer. However, mediastinoscopy is at present still regarded as the gold standard in the region of the anterior mediastinum since EUS can not image this region due to the air filled trachea. Recently, endobronchial ultrasound guided transbronchial needle aspiration Biopsy (EBUS-TBNA) has been developed and several publications have now documented high diagnostic values with sensitivities of more than 90% in the staging of NSCLC. A recent publication from our group has documented a sensitivity and specificity of 100% when EUS-FNA and EBUS-TBNA is used in combination for staging of the mediastinum. It seems therefore logical to assume that the combination of EUS-FNA and EBUS-TBNA will replace more invasive methods such as mediastinoscopy for diagnosis and staging of lung cancers in the near future. PMID- 17921948 TI - [Non invasive methods in competition for EUS (US, CE-US, CT, MRI and PET/CT): the radiologist point of view]. AB - Recent technological improvements such as spiral CT, CPMR, PET-CT, US and MRI specific contrast agents have increased the accuracy of non-invasive exams performed by radiologists. US is often the first step of the examination. MRI and PET-CT are considered supplementary. At present, CT is the method of choice for the diagnosis, staging and therapeutic planning of pancreatic tumors for which a multidisciplinary approach is however mandatory. PMID- 17921949 TI - [Role of EUS in a diagnostic and staging algorithm of pancreatic carcinoma: the surgeon point of view]. AB - Survival of pancreatic cancer is improved by surgery and is related to R0 resection. An accurate diagnosis and a careful staging are mandatory. Differential diagnosis must be estabilished between the different pancreatic lesions as carcinoma, chronic pancreatitis, cystic or endocrine neoplasms. Endoscopic ultrasound (EUS) is the best technique for diagnosis and allows cytological examination by fine needle aspiration (FNA). Preoperative resectability is defined by EUS in borderline tumors. EUS is a useful procedure for the surgical strategy of pancreatic cancer. PMID- 17921950 TI - Endoscopic ultrasound and fine needle aspiration in inflammatory and cystic pancreatic pathology. AB - To achieve an accurate diagnosis on the nature of pancreatic cystic lesions may be difficult. This review article attempts to provide an overview of the potential role of endoscopic ultrasound (EUS) and EUS guided fine needle aspiration in the evaluation of cystic lesions of the pancreas. PMID- 17921951 TI - Role of endoscopic ultrasound in biliary and unexplained pancreatitis. AB - In the Western world, gallstone disease accounts for approximately half of the cases of acute pancreatitis. The benefit of therapeutic endoscopic retrograde cholangiopancreatography (ERCP) has been clearly proven for this disease. On the basis of history, laboratory, and noninvasive imaging studies, about 10-30% of patients with acute pancreatitis have no apparent aetiology. This is critical because it limits the optimal therapeutic management and the prevention of recurrence. Endoscopic ultrasound (EUS) has emerged as an important procedure to clarify the aetiology of acute pancreatitis. After extensive investigation that included EUS, 8-20% of episodes remained idiopathic. Moreover, EUS carried minimal risk when compared to ERCP. The present short review attempts to update on the indications for the employment of EUS in the diagnosis of biliary stones as well as in the clarification of aetiology in the case of unexplained pancreatitis. PMID- 17921952 TI - [EUS-FNA cytology of pancreatic exocrine tumors. Comparison of experiences with pathological diagnosis]. AB - In this minireview, the main clinical applications and results of EUS-FNA (Endoscopic Ultrasound-Guided Fine Needle Aspiration) Cytology in the cystic and solid neoplastic lesions of the exocrine pancreas are described. EUS-FNA provides a safe and accurate mean to diagnose pancreatic tumors in early and advanced stages. A personal observation of a case of acinic cell carcinoma is briefly presented, with extensive cytological iconography of routinely stained smears, integrated with cytochemical/immunocytochemical analysis for diagnostic purposes. PMID- 17921953 TI - [Histological sectioning of brush bristles allows an improved diagnosis of biliary tract lesions]. AB - Biliary tract brush cytology is increasingly being recognized as a favoured method for evaluating abnormalities of the biliary tract. In order to increase the diagnostic accuracy, we devise a new brush processing method finalized to the complete and ideal cytologic examination of the collected material. Small fragments of the mucosa, of inflammatory cell aggregates or of carcinomas are observed and the results are optimally fixed and allow a definitive histological diagnosis. PMID- 17921954 TI - Biliary therapy: are we ready for EUS-guidance? AB - Endoscopic retrograde cholangiopancreaticography is the standard therapy for the therapy of biliary obstruction. However, the success rate is not 100%, depending on various patient and physician related factors. In these cases, where endoscopic drainage is not possible, either percutaneous drainage or surgery are established alternatives. Both modalities carry a higher complication rate and are more invasive than endoscopic drainage. With linear echo-endoscopes, left intrahepatic bile ducts as well as the distal common bile duct can be visualized from the stomach or the duodenal bulb respectively. This opens up the possibility of puncturing the bile ducts under real time ultrasound control from the intestinal lumen. There are two different techniques to achieve biliary drainage after gaining EUS guided access: The first is direct biliary drainage in the intestinal lumen by placing a stent through the wall of the stomach/duodenum after placement of a guidewire through an 19gauge needle into the biliary tract. This technique usually requires some form of bouginage once the guide wire has been placed and is very similar to EUS guided pseudocyst drainage. The second technique is the rendezvous technique, where the guidewire is manipulated through the stricture and the papilla. Thereafter the wire is captured with a standard duodenoscope and a biliary drainage is performed through the papilla in established fashion. With both techniques fluoroscopic control in addition to EUS is needed. So far both techniques have been described in case reports and small series only. Large prospective series as well as controlled trials that compare EUS guided techniques with ERCP or PTC are lacking. The most common complication is biliary leakage, especially if direct drainage is performed. Other common complications include cholangitis, stent migration and occlusion as well as pain. As long as large prospective series are lacking, EUS guided biliary drainage should be restricted to selected patients where ERCP has repeatedly failed or is impossible due to surgically altered anatomy. Furthermore this technically demanding procedure should be performed only in centres with extensive experience in linear EUS and therapeutic biliary ERCP. The possible advantages over percutaneous drainage like patient comfort and morbidity have to be proven in randomized trials. PMID- 17921955 TI - Mutations in gastrointestinal stromal tumors diagnosed by endoscopic ultrasound guided fine needle aspiration. AB - Gastrointestinal stromal tumors (GISTs) are characterized by overexpression and mutations of c-Kit. Approximately 80% of c-Kit mutations occur in exon 11, being a response factor to imatinib (Gleevec) therapy. We aimed to assess whether c-Kit and PDGFRA mutation analysis of GISTs obtained by endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) could be routinely performed. Mutation analysis of c-Kit hotspot exons (9, 11, 13 and 17) and PDGFRA hotspot exons (12 and 18) was performed in aspirates 51 mesenchymal tumors. We identified c-Kit mutations in 61% of GIST cases, in accordance with previously published ranges (30-90%). Nearly 95% (19/20) of c-kit-mutant tumors carried exon 11 mutations. Mutation analysis is possible in FNA cell blocks and can assist in the diagnosis and therapeutic decisions in GIST cases. PMID- 17921956 TI - Endoscopic ultrasonography in submucosal lesions and extrinsic compressions of the gastrointestinal tract. AB - Endoscopic ultrasonography (EUS) provides an accurate exploration of the wall of the digestive organs, which makes it the procedure of choice for the diagnosis of submucosal lesions of the gastrointestinal (GI) tract. It reveals the exact origin of a lesion, whether it be inside or outside the gastrointestinal wall, and allows to define the characteristics of the lesion and help us to know its nature and make possible to perform EUS-guided fine-needle aspiration (EUS-FNA) to cytology with immunohistochemial analysis. PMID- 17921957 TI - [On-site evaluation and triage for endoscopic ultrasound-guided fine needle aspiration cytology. The Turin experience]. AB - AIM: Evaluation of the importance of the on-site presence of a skilled cytopathologist during endoscopic ultrasound-guided fine needle aspiration at determining samples' adequacy and performing ancillary techniques which can be helpful for the diagnosis. METHODS: A retrospective analysis of our institute's experience with EUS-FNA sampling is presented. From January 2001 to May 2007, 404 patients underwent the EUS-FNA evaluation. From 2003 a cytopathologist was present during the procedure and started making an extemporary evaluation of the samples' adequacy. RESULTS: Before 2003, a final cytological diagnosis was available in only 70% of the cases (without an on-site cytopathologist). After 2003, in 90% of the cases (with an on-site cytopathologist). It is possible planning and performing: immunocytochemistry on cell block material including evaluation of the proliferation index; to obtain a sample for the flow cytometry in cases of lymphomas or a microbiologic workup in cases of infective lesions. CONCLUSION: The quality of the specimens and the proper handling of the aspirated sample are very important to succesfully obtain a definitive cytological diagnosis in EUS-FNA. On-site evaluation and triage of the material allow to improve the accuracy of the diagnosis. PMID- 17921958 TI - State of the art review: developments in endoscopic ultrasound in esophageal and gastric disease in 2005 and 2006. AB - The subject of this review will be recent developments in the field of EUS in esophageal and gastric disease. Selected articles published in the English language in 2005, 2006 and the first half of 2007 will be reviewed in more detail. Subjects discussed in this paper will be esophageal cancer and EUS with special emphasis on staging accuracy, the value of EUS in restaging after neo adjuvant treatment and quality and impact on the management of patients. Furthermore papers with the subjects gastric cancer and submucosal tumors will be discussed. PMID- 17921959 TI - Searching a role for endoscopic ultrasonography in Barrett's esophageus and other acid-related or gastrointestinal motility disorders. AB - Endoscopic ultrasonography (EUS) can image the GI-wall in detail. Substaging of T1-tumours (e.g. in Barrett's esophageus) is possible with EUS which may have clinical significance regarding therapeutic options. Abnormal motility occurs in several gastrointestinal (GI) diseases and EUS can be used in biomechanical and motility studies. Three-dimensional EUS, elastography and strain rate imaging are new techniques which are promising in imaging the GI-tract and in the examination of gut motility. PMID- 17921960 TI - Endoscopic mucosal resection-endoscopic submucosal dissection: do we really need endoscopic ultrasonography assistance? AB - Endoscopic mucosal resection has become the standard of care for early gastrointestinal cancer. The application of this new treatment requires an accurate stadiation of the neoplasia. The exclusion of nodal involvement and the evaluation of the depth of tumor penetration within the gastrointestinal wall is essential to select patients who can benefit from this approach. Echoendoscopy allows endoscopists to evaluate subtle changes in the layers of the gastrointestinal wall giving an important aid to local tumor staging and planning the adequate treatment. PMID- 17921961 TI - Mediastinal and gastric EUS: indications and technique of examination. AB - Endoscopic ultrasound (EUS) is a minimally invasive technology using a high frequency ultrasound transducer that is incorporated into the tip of a conventional endoscope. This technique permits the diagnosis and staging of gastrointestinal lesions and mediastinal mass. EUS-guided fine-needle aspiration (FNA) enables operators to obtain tissue diagnosis of most visualized lesions. This paper reports on the technique of examination and indications for mediastinal and gastric EUS. PMID- 17921962 TI - Endoscopic ultrasonography (EUS) in portal hypertension. AB - Although the first studies on EUS in portal hypertension are by now dated approximately 20 years ago, this method still has not become a routine examination among the diagnostic and therapeutic examinations in this field. EUS has an important role in the following applications: early diagnosis of esophageal and gastric varices and of hypertensive gastropathy, screening of subjects to treat medically, endoscopically or surgically, and their follow-up, EUS-guided injection therapy of varices. Several papers have confirmed and/or added important clinical applications to the EUS use in such patients. In fact, if at the beginnig the studies were concentrated on the best way to show varices compared to the endoscopic examination, or on aspects still unknown such as the presence of periesophageal veins and collaterals increased in number and diameter, at present sophisticated instruments with several types of Doppler and therapeutic possibilities are available. The observation of perforating veins under the esophagogastric junction seems to have a particular clinical interest having a predictive value on the effectiveness of the sclerotherapy and therefore on the final eradication. Also the several instrumentations used can modify the RESULTS: Using the high frequencies EUS miniprobes, the sensibility in identifying even the minimal or initial varices increases. In the evaluation of the pharmacological therapy, EUS with the new instruments equipped with color Doppler can be very useful and can replace other diagnostic methods very invasive and dangerous and thus used only in highly specialized centers. As to the nursing aspect of EUS in portal hypertension, it must be underlined that such pathology is often the consequence of viral hepatitis infection. Therefore, beside a correct clinical evaluationl before endoscopic ultrasound examination, all devices, drugs and anything else could be be necessary to complete with success an intervention, e.g. sclerotherapy in emergency, should be prepared in advance. PMID- 17921963 TI - Europa Medicophysica, the European and Mediterranean evidence-based clinical Journal of Physical and Rehabilitation Medicine. PMID- 17921964 TI - A systematic review of physical and rehabilitation medicine topics as developed by the Cochrane Collaboration. AB - The aim of this paper is to give an overview of the Cochrane Collaboration, its development over the years, and its usefulness for physical and rehabilitation medicine specialists. After introducing the Cochrane Collaboration, we systematically analyzed all titles of the reviews in each of the 50 review groups. For ''Rehabilitation,'' we included not only exercise and physical therapy, but also all of the educational and pharmacological interventions aimed at supporting rehabilitation. The search was performed using Issue 2, 2007 of the Cochrane Library. We retrieved 138 completed reviews that dealt with rehabilitation interventions performed by 20 review groups. No umbrella reviews could be found at present. The most prolific groups in the rehabilitation field were: musculoskeletal (28 reviews), stroke (20), back (18), and the movement disorder group (13). The most discussed intervention was exercise (37 reviews), dealing with physical medicine (20) and pharmacological intervention (11). Six other reviews dealt with multidisciplinary rehabilitation. Low back pain treatment and stroke were the most investigated topics. The Cochrane Collaboration serves today as a main step in increasing an evidence-based approach to medicine and rehabilitation. Europa Medicophysica is continuously increasing its importance and presence in the world of physical and rehabilitation medicine, and part of its mission is to increase and strengthen an evidence-based approach to this field. We will continue to look at the contents of the Cochrane database and regularly report on its updates concerning topics of interest for physical and rehabilitation medicine. PMID- 17921965 TI - Ergonomic and physiotherapeutic interventions for treating work-related complaints of the arm, neck or shoulder in adults. A Cochrane systematic review. AB - BACKGROUND: Conservative interventions such as physiotherapy and ergonomic adjustments (such as keyboard adjustments or ergonomic advice) play a major role in the treatment of most work-related complaints of the arm, neck or shoulder (CANS). Objectives. This systematic review aims to determine whether conservative interventions have a significant impact on outcomes for work-related CANS in adults. Search strategy. We searched the Cochrane Bone, Joint and Muscle Trauma Group Specialised Register (March 2005) and Cochrane Rehabilitation and Related Therapies Field Specialised Register (March 2005), the Cochrane Controlled Trials Register (The Cochrane Library, Issue 1, 2005), PubMed, EMBASE, CINAHL, AMED and reference lists of articles. The date of the last search was March 2005. No language restrictions were applied. Selection criteria. We included randomised controlled trials and concurrent controlled trials studying conservative interventions (e.g. exercises, relaxation, physical applications, biofeedback, myofeedback and work-place adjustments) for adults suffering CANS. Data collection and analysis. Two authors independently selected trials from the search yield, assessed the methodological quality using the Delphi list, and extracted relevant data. We pooled data or, in the event of clinical heterogeneity or lack of data, we used a rating system to assess levels of evidence. MAIN RESULTS: For this update we included six additional studies; twenty-one trials in total. Seventeen trials included people with chronic nonspecific neck or shoulder complaints, or nonspecific upper extremity disorders. Over twenty-five interventions were evaluated; six main subgroups of interventions could be determined: exercises, manual therapy, massage, ergonomics, energised splint and individual treatment versus group therapy. Overall, the quality of the studies was poor. In 14 studies a form of exercise was evaluated, and contrary to the previous review we now found limited evidence about the effectiveness of exercises when compared to massage and conflicting evidence when exercises are compared to no treatment. In this update there is limited evidence for adding breaks during computer work; massage as add-on treatment on manual therapy, manual therapy as add-on treatment on exercises; and some keyboard designs when compared to other keyboards or placebo in participants with carpal tunnel syndrome. CONCLUSIONS: There is limited evidence for the effectiveness of keyboards with an alternative force-displacement of the keys or an alternative geometry, and limited evidence for the effectiveness of exercises compared to massage, breaks during computer work compared to no breaks; massage as an add-on treatment to manual therapy, and manual therapy as an add-on treatment to exercises. PMID- 17921966 TI - Rehabilitation and outcome measurement: where is Rasch analysis-going? AB - Outcomes are clinical or functional changes at the whole-person level. Measures are based on questionnaires which summate scores assigned to a series of items representing a person's variable. The metric properties of raw scores are known to be of limited validity. Yet, such variables represent the main target of outcome assessment in Rehabilitation Medicine as far as they may cover performances, perceptions and knowledge. Rasch statistical models, first available in English to a large readesrship in late 70s, allow to transform the arbitary raw scores into true linear measures. Rehabilitation seemed a privileged field for the application of Rasch models, yet these are still far from becoming popular. A bibliometric study was conducted across ten leading digital libraries. A MEDLINE search showed a dramatic increment of published papers covering the intersection between the MESH terms ''Rehabilitation'' and ''Outcome assessment'', which rose in from 5 to 4 302 between the 1981-90 and 2001-07 time frames, respectively. ''Rasch'' paper also rose remarkably from 35 to 539. When the triple intersection was considered, articles only rose from 0 to 12. Results point towards a privileged application of Rasch analysis to build and refine questionnaires, rather than to actually measuring people. Commentaries and suggestions are invited. PMID- 17921967 TI - PubMed at its best. Useful skills to perform an effective search, save it and automatically receive updates. PMID- 17921968 TI - Are current staging systems for cancer serving us well? PMID- 17921970 TI - Conservative management of testicular germ-cell tumors. AB - Cure rates of germ-cell cancer have been greater than 95% for the last decade; therefore, over the past few years there has been a greater focus on 'conservative' approaches to treatment. The increased incidence of late non-germ cell cancers in the era of radiotherapy and the incidence of subclinical testicular deficiency and metabolic syndrome in cured patients have accelerated this trend. Taking account of the increase in cure rates of primary chemotherapy failures from 5% to 60% with intensification of chemotherapy and surgery, this Review focuses on three areas: lessons from the initial failed trials of less intensive treatment (i.e. bleomycin withdrawal and carboplatin substitution) that emphasised the need for improved salvage therapy; successes of reducing treatment of patients with metastases classed as good-risk from four cycles(20 days) to three cycles(9 days) and using 1 day carboplatin instead of 21 day radiotherapy as adjuvant for stage 1 seminoma; and the unexpected finding at 5 years of a 72% reduction of contralateral second germ-cell cancer. This finding provides the stimulus for the next generation of conservative trials using organ preservation, aiming to reduce occurrence of metabolic syndrome and using new radiological and minimal surgery techniques to accelerate the assessment of less toxic drugs and new approaches for combined medial and surgical treatment. PMID- 17921969 TI - The role of hexaminolevulinate fluorescence cystoscopy in bladder cancer. AB - Non-muscle-invasive bladder cancer is labor intensive and costly to manage. Owing to long-term survival rates and life-long monitoring and treatment, it is the most expensive cancer to manage in per-patient terms. Significant costs are attributable to the treatment of recurrences and complications. Fluorescence guided cystoscopy, using 5-aminolevulinic acid (ALA) or its hexyl ester, hexaminolevulinate 5-ALA (Hexvix [HAL], Photocure, Oslo, Norway), improves the detection of bladder tumors, particularly carcinoma in situ, compared with standard white-light cystoscopy. The quality of transurethral resection of the bladder tumor is also improved. It has been shown that improved tumor detection leads to better patient management and, in the case of ALA, reduced long-term recurrence rates and costs. Long-term studies in this area with HAL are ongoing. The technique is well tolerated and is a useful adjunct to white-light cystoscopy. PMID- 17921971 TI - Surgery Insight: optimizing open nerve-sparing radical prostatectomy techniques for improved outcomes. AB - Men with prostate cancer have a variety of treatment choices available, including expectant management with deferred treatment, brachytherapy, external-beam radiation therapy, or both of the latter options--with or without hormonal therapy, cryotherapy, and radical prostatectomy (RP). Physicians have long endeavored to guide patients through these choices on the basis of the health threat posed by the cancer, the potential effectiveness and complications associated with treatment, and the patient's life expectancy. As early detection programs now identify cancers much earlier in their natural history, individual patients have a longer life expectancy than in the past. The patient and physician must, therefore, weigh the potential benefit of the selected treatment with the risk of early or delayed complications that would detract from the patient's quality of life. Optimally, when a surgical approach is used to treat prostate cancer, the operation removes the cancer completely with negative surgical margins, avoids excessive blood loss or serious perioperative complications, and culminates in complete recovery of continence and potency. To achieve this, the surgeon must treat sufficient periprostatic tissue to achieve cure while preserving the cavernosal nerves required for erectile function and the neuromusculature required for normal urinary and bowel function. Evidence suggests that the small details of how a surgery is performed have a major impact on the outcome of RP. Here the role of surgical techniques in determining oncologic and quality of life outcomes after RP, focusing on open RP, are presented. PMID- 17921972 TI - Urolithiasis: the influence of stone size on management. AB - BACKGROUND: A 37-year-old female presented with pain in the right lower lateral abdomen. She had experienced a ureteral stone in the past. INVESTIGATIONS: Serum electrolyte levels, blood urea nitrogen, serum creatinine level, full blood count, urinalysis, CT. DIAGNOSIS: Urolithiasis. MANAGEMENT: Cystoscopy and right retrograde pyelogram. PMID- 17921973 TI - Roadmap for revitalization? PMID- 17921974 TI - Industry concerns mount over CIRM grants. PMID- 17921975 TI - Europe's anti-GM stance to presage animal feed shortage? PMID- 17921976 TI - Death in gene therapy trial raises questions about private IRBs. PMID- 17921977 TI - Nanotech under scrutiny, but tighter regulation remains distant. PMID- 17921979 TI - Profile: Alpheus Bingham. PMID- 17921981 TI - Mice with a human touch. PMID- 17921983 TI - A shocking biodefense editorial. PMID- 17921984 TI - The FDA animal efficacy rule and biodefense. PMID- 17921985 TI - Conserving biodiversity using patent law. PMID- 17921986 TI - Why Cabilly must stand. PMID- 17921987 TI - European regulation tackles tissue engineering. PMID- 17921988 TI - The power and pitfalls of outsourcing. PMID- 17921989 TI - What's fueling the biotech engine? PMID- 17921990 TI - Obviousness, hindsight and perspective: the impact of KSR v. Teleflex on biotech and pharmaceutical patents. PMID- 17921991 TI - Recent patent applications in protein purification. PMID- 17921992 TI - No free energy lunch. PMID- 17921993 TI - Network pharmacology. PMID- 17921994 TI - AAV vectors and tumorigenicity. PMID- 17921995 TI - Many ways to pluripotency. PMID- 17921997 TI - Drug-target network. AB - The global set of relationships between protein targets of all drugs and all disease-gene products in the human protein-protein interaction or 'interactome' network remains uncharacterized. We built a bipartite graph composed of US Food and Drug Administration-approved drugs and proteins linked by drug-target binary associations. The resulting network connects most drugs into a highly interlinked giant component, with strong local clustering of drugs of similar types according to Anatomical Therapeutic Chemical classification. Topological analyses of this network quantitatively showed an overabundance of 'follow-on' drugs, that is, drugs that target already targeted proteins. By including drugs currently under investigation, we identified a trend toward more functionally diverse targets improving polypharmacology. To analyze the relationships between drug targets and disease-gene products, we measured the shortest distance between both sets of proteins in current models of the human interactome network. Significant differences in distance were found between etiological and palliative drugs. A recent trend toward more rational drug design was observed. PMID- 17921998 TI - The Functional Genomics Experiment model (FuGE): an extensible framework for standards in functional genomics. AB - The Functional Genomics Experiment data model (FuGE) has been developed to facilitate convergence of data standards for high-throughput, comprehensive analyses in biology. FuGE models the components of an experimental activity that are common across different technologies, including protocols, samples and data. FuGE provides a foundation for describing entire laboratory workflows and for the development of new data formats. The Microarray Gene Expression Data society and the Proteomics Standards Initiative have committed to using FuGE as the basis for defining their respective standards, and other standards groups, including the Metabolomics Standards Initiative, are evaluating FuGE in their development efforts. Adoption of FuGE by multiple standards bodies will enable uniform reporting of common parts of functional genomics workflows, simplify data integration efforts and ease the burden on researchers seeking to fulfill multiple minimum reporting requirements. Such advances are important for transparent data management and mining in functional genomics and systems biology. PMID- 17921999 TI - From XenoMouse technology to panitumumab, the first fully human antibody product from transgenic mice. AB - Therapeutic monoclonal antibodies have shown limited efficacy and safety owing to immunogenicity of mouse sequences in humans. Among the approaches developed to overcome these hurdles were transgenic mice genetically engineered with a 'humanized' humoral immune system. One such transgenic system, the XenoMouse, has succeeded in recapitulating the human antibody response in mice, by introducing nearly the entire human immunoglobulin loci into the germ line of mice with inactivated mouse antibody machinery. XenoMouse strains have been used to generate numerous high-affinity, fully human antibodies to targets in multiple disease indications, many of which are progressing in clinical development. However, validation of the technology has awaited the recent regulatory approval of panitumumab (Vectibix), a fully human antibody directed against epidermal growth factor receptor (EGFR), as treatment for people with advanced colorectal cancer. The successful development of panitumumab represents a milestone for mice engineered with a human humoral immune system and their future applications. PMID- 17922002 TI - Nucleation and growth mechanism of ferroelectric domain-wall motion. AB - The motion of domain walls is critical to many applications involving ferroelectric materials, such as fast high-density non-volatile random access memory. In memories of this sort, storing a data bit means increasing the size of one polar region at the expense of another, and hence the movement of a domain wall separating these regions. Experimental measurements of domain growth rates in the well-established ferroelectrics PbTiO3 and BaTiO3 have been performed, but the development of new materials has been hampered by a lack of microscopic understanding of how domain walls move. Despite some success in interpreting domain-wall motion in terms of classical nucleation and growth models, these models were formulated without insight from first-principles-based calculations, and they portray a picture of a large, triangular nucleus that leads to unrealistically large depolarization and nucleation energies. Here we use atomistic molecular dynamics and coarse-grained Monte Carlo simulations to analyse these processes, and demonstrate that the prevailing models are incorrect. Our multi-scale simulations reproduce experimental domain growth rates in PbTiO3 and reveal small, square critical nuclei with a diffuse interface. A simple analytic model is also proposed, relating bulk polarization and gradient energies to wall nucleation and growth, and thus rationalizing all experimental rate measurements in PbTiO3 and BaTiO3. PMID- 17922003 TI - Coupling of cortical dynein and G alpha proteins mediates spindle positioning in Caenorhabditis elegans. AB - Despite being essential for spatial cell division control, the mechanisms governing spindle positioning remain incompletely understood. In the Caenorhabditis elegans one-cell stage embryo, the spindle becomes asymmetrically positioned during anaphase through the action of as-yet unidentified cortical force generators that pull on astral microtubules and that depend on two G alpha proteins and associated proteins. We performed spindle-severing experiments following temporally restricted gene inactivation and drug exposure, and established that microtubule dynamics and dynein are both required for generating efficient pulling forces. We found that the G alpha-associated proteins GPR-1/2 and LIN-5 interact in vivo with LIS-1, a component of the dynein complex. Moreover, we discovered that the LIN-5, GPR-1/2 and the G alpha proteins promote the presence of the dynein complex at the cell cortex. Our findings suggest a mechanism by which the G alpha proteins enable GPR-1/2 and LIN-5 recruitment to the cortex, thus ensuring the presence of cortical dynein. Together with microtubule dynamics, this allows pulling forces to be exerted and proper cell division to be achieved. PMID- 17922004 TI - SNARE proteins mediate fusion between cytosolic lipid droplets and are implicated in insulin sensitivity. AB - The accumulation of cytosolic lipid droplets in muscle and liver cells has been linked to the development of insulin resistance and type 2 diabetes. Such droplets are formed as small structures that increase in size through fusion, a process that is dependent on intact microtubules and the motor protein dynein. Approximately 15% of all droplets are involved in fusion processes at a given time. Here, we show that lipid droplets are associated with proteins involved in fusion processes in the cell: NSF (N-ethylmaleimide-sensitive-factor), alpha-SNAP (soluble NSF attachment protein) and the SNAREs (SNAP receptors), SNAP23 (synaptosomal-associated protein of 23 kDa), syntaxin-5 and VAMP4 (vesicle associated membrane protein 4). Knockdown of the genes for SNAP23, syntaxin-5 or VAMP4, or microinjection of a dominant-negative mutant of alpha-SNAP, decreases the rate of fusion and the size of the lipid droplets. Thus, the SNARE system seems to have an important role in lipid droplet fusion. We also show that oleic acid treatment decreases the insulin sensitivity of heart muscle cells, and this sensitivity is completely restored by transfection with SNAP23. Thus, SNAP23 might be a link between insulin sensitivity and the inflow of fatty acids to the cell. PMID- 17922005 TI - ATP-competitive inhibitors of the mitotic kinesin KSP that function via an allosteric mechanism. AB - The mitotic kinesin KSP (kinesin spindle protein, or Eg5) has an essential role in centrosome separation and formation of the bipolar mitotic spindle. Its exclusive involvement in the mitotic spindle of proliferating cells presents an opportunity for developing new anticancer agents with reduced side effects relative to antimitotics that target tubulin. Ispinesib is an allosteric small molecule KSP inhibitor in phase 2 clinical trials. Mutations that attenuate ispinesib binding to KSP have been identified, which highlights the need for inhibitors that target different binding sites. We describe a new class of selective KSP inhibitors that are active against ispinesib-resistant forms of KSP. These ATP-competitive KSP inhibitors do not bind in the nucleotide binding pocket. Cumulative data from generation of resistant cells, site-directed mutagenesis and photo-affinity labeling suggest that they compete with ATP binding via a novel allosteric mechanism. PMID- 17922006 TI - Figure-ground mechanisms provide structure for selective attention. AB - Attention depends on figure-ground organization: figures draw attention, whereas shapes of the ground tend to be ignored. Recent research has revealed mechanisms for figure-ground organization in the visual cortex, but how these mechanisms relate to the attention process remains unclear. Here we show that the influences of figure-ground organization and volitional (top-down) attention converge in single neurons of area V2 in Macaca mulatta. Although we found assignment of border ownership for attended and for ignored figures, attentional modulation was stronger when the attended figure was located on the neuron's preferred side of border ownership. When the border between two overlapping figures was placed in the receptive field, responses depended on the side of attention, and enhancement was generally found on the neuron's preferred side of border ownership. This correlation suggests that the neural network that creates figure-ground organization also provides the interface for the top-down selection process. PMID- 17922007 TI - Transcriptional repression coordinates the temporal switch from motor to serotonergic neurogenesis. AB - In many regions of the developing CNS, distinct cell types are born at different times. The means by which discrete and stereotyped temporal switches in cellular identities are acquired remains poorly understood. To address this, we have examined how visceral motor neurons (VMNs) and serotonergic neurons, two neuronal subtypes, are sequentially generated from a common progenitor pool in the vertebrate hindbrain. We found that the forkhead transcription factor Foxa2, acting in progenitors, is essential for the transition from VMN to serotonergic neurogenesis. Loss-of-function and gain-of-function experiments indicated that Foxa2 activates the switch through a temporal cross-repressive interaction with paired-like homeobox 2b (Phox2b), the VMN progenitor determinant. This mechanism bears a marked resemblance to the cross-repression between neighboring domains of transcription factors that establish discrete progenitor identities along the spatial axes. Moreover, the subsequent differentiation of central serotonergic neurons required both the suppression of VMN neurogenesis and the induction of downstream intrinsic determinants of serotonergic identity by Foxa2. PMID- 17922008 TI - Sensory processing in the Drosophila antennal lobe increases reliability and separability of ensemble odor representations. AB - Here we describe several fundamental principles of olfactory processing in the Drosophila melanogaster antennal lobe (the analog of the vertebrate olfactory bulb), through the systematic analysis of input and output spike trains of seven identified glomeruli. Repeated presentations of the same odor elicit more reproducible responses in second-order projection neurons (PNs) than in their presynaptic olfactory receptor neurons (ORNs). PN responses rise and accommodate rapidly, emphasizing odor onset. Furthermore, weak ORN inputs are amplified in the PN layer but strong inputs are not. This nonlinear transformation broadens PN tuning and produces more uniform distances between odor representations in PN coding space. In addition, portions of the odor response profile of a PN are not systematically related to their direct ORN inputs, which probably indicates the presence of lateral connections between glomeruli. Finally, we show that a linear discriminator classifies odors more accurately using PN spike trains than using an equivalent number of ORN spike trains. PMID- 17922009 TI - STAT5A is epigenetically silenced by the tyrosine kinase NPM1-ALK and acts as a tumor suppressor by reciprocally inhibiting NPM1-ALK expression. AB - Although STAT5A and STAT5B have some nonredundant functional properties, their distinct contributions to carcinogenesis are not clearly defined. Here we report that STAT5A expression is selectively inhibited by DNA methylation of the STAT5A gene promoter region in cells expressing the oncogenic tyrosine kinase NPM1-ALK (also known as NPM-ALK). The DNA methylation is induced by NPM1-ALK itself via STAT3, and is associated with binding to the promoter of the gene encoding MeCP2 capping protein and with lack of binding of the STAT5A gene transcription activator SP1. Reversal of methylation by the DNA methyltransferase inhibitor 5' aza-2'-deoxycytidine restores SP1 binding and STAT5A gene expression. Notably, the induced or exogenously expressed STAT5A protein binds to the enhancer and intron 14 of the NPM1-ALK gene and triggers selective suppression of NPM1-ALK expression. These results show that NPM1-ALK induces epigenetic silencing of STAT5A gene and that STAT5A protein can act as a key tumor suppressor by reciprocally inhibiting expression of NPM1-ALK. PMID- 17922010 TI - Deacetylase inhibition promotes the generation and function of regulatory T cells. AB - Histone/protein deacetylases (HDACs) regulate chromatin remodeling and gene expression as well as the functions of more than 50 transcription factors and nonhistone proteins. We found that administration of an HDAC inhibitor (HDACi) in vivo increased Foxp3 gene expression, as well as the production and suppressive function of regulatory T cells (T(reg) cells). Although T(reg) cells express multiple HDACs, HDAC9 proved particularly important in regulating Foxp3-dependent suppression. Optimal T(reg) function required acetylation of several lysines in the forkhead domain of Foxp3, and Foxp3 acetylation enhanced binding of Foxp3 to the Il2 promoter and suppressed endogenous IL-2 production. HDACi therapy in vivo enhanced T(reg)-mediated suppression of homeostatic proliferation, decreased inflammatory bowel disease through T(reg)-dependent effects, and, in conjunction with a short course of low-dose rapamycin, induced permanent, T(reg)-dependent cardiac and islet allograft survival and donor-specific allograft tolerance. Our data show that use of HDACi allows the beneficial pharmacologic enhancement of both the numbers and suppressive function of Foxp3(+) T(reg) cells. PMID- 17922011 TI - Conformational dynamics of the KcsA potassium channel governs gating properties. AB - K+ channels conduct and regulate K+ flux across the cell membrane. Several crystal structures and biophysical studies of tetrameric ion channels have revealed many of the structural details of ion selectivity and gating. A narrow pore lined with four arrays of carbonyl groups is responsible for ion selectivity, whereas a conformational change of the four inner transmembrane helices (TM2) is involved in gating. We used NMR to examine full-length KcsA, a prototypical K+ channel, in its open, closed and intermediate states. These studies reveal that at least two conformational states exist both in the selectivity filter and near the C-terminal ends of the TM2 helices. In the ion conducting open state, we observed rapid structural exchange between two conformations of the filter, presumably of low and high K+ affinity, respectively. Such measurements of millisecond-timescale dynamics reveal the basis for simultaneous ion selection and gating. PMID- 17922012 TI - Molecular driving forces determining potassium channel slow inactivation. AB - K+ channels undergo a time-dependent slow inactivation process that plays a key role in modulating cellular excitability. Here we show that in the prokaryotic proton-gated K+ channel KcsA, the number and strength of hydrogen bonds between residues in the selectivity filter and its adjacent pore helix determine the rate and extent of C-type inactivation. Upon channel activation, the interaction between residues at positions Glu71 and Asp80 promotes filter constriction parallel to the permeation pathway, which affects K+-binding sites and presumably abrogates ion conduction. Coupling between these two positions results in a quantitative correlation between their interaction strength and the stability of the inactivated state. Engineering of these interactions in the eukaryotic voltage-dependent K+ channel Kv1.2 suggests that a similar mechanistic principle applies to other K+ channels. These observations provide a plausible physical framework for understanding C-type inactivation in K+ channels. PMID- 17922013 TI - Ancestral reconstruction of segmental duplications reveals punctuated cores of human genome evolution. AB - Human segmental duplications are hotspots for nonallelic homologous recombination leading to genomic disorders, copy-number polymorphisms and gene and transcript innovations. The complex structure and history of these regions have precluded a global evolutionary analysis. Combining a modified A-Bruijn graph algorithm with comparative genome sequence data, we identify the origin of 4,692 ancestral duplication loci and use these to cluster 437 complex duplication blocks into 24 distinct groups. The sequence-divergence data between ancestral-derivative pairs and a comparison with the chimpanzee and macaque genome support a 'punctuated' model of evolution. Our analysis reveals that human segmental duplications are frequently organized around 'core' duplicons, which are enriched for transcripts and, in some cases, encode primate-specific genes undergoing positive selection. We hypothesize that the rapid expansion and fixation of some intrachromosomal segmental duplications during great-ape evolution has been due to the selective advantage conferred by these genes and transcripts embedded within these core duplications. PMID- 17922014 TI - Network modeling links breast cancer susceptibility and centrosome dysfunction. AB - Many cancer-associated genes remain to be identified to clarify the underlying molecular mechanisms of cancer susceptibility and progression. Better understanding is also required of how mutations in cancer genes affect their products in the context of complex cellular networks. Here we have used a network modeling strategy to identify genes potentially associated with higher risk of breast cancer. Starting with four known genes encoding tumor suppressors of breast cancer, we combined gene expression profiling with functional genomic and proteomic (or 'omic') data from various species to generate a network containing 118 genes linked by 866 potential functional associations. This network shows higher connectivity than expected by chance, suggesting that its components function in biologically related pathways. One of the components of the network is HMMR, encoding a centrosome subunit, for which we demonstrate previously unknown functional associations with the breast cancer-associated gene BRCA1. Two case-control studies of incident breast cancer indicate that the HMMR locus is associated with higher risk of breast cancer in humans. Our network modeling strategy should be useful for the discovery of additional cancer-associated genes. PMID- 17922015 TI - Development of plasmacytoid and conventional dendritic cell subtypes from single precursor cells derived in vitro and in vivo. AB - The development of functionally specialized subtypes of dendritic cells (DCs) can be modeled through the culture of bone marrow with the ligand for the cytokine receptor Flt3. Such cultures produce DCs resembling spleen plasmacytoid DCs (pDCs), CD8(+) conventional DCs (cDCs) and CD8(-) cDCs. Here we isolated two sequential DC-committed precursor cells from such cultures: dividing 'pro-DCs', which gave rise to transitional 'pre-DCs' en route to differentiating into the three distinct DC subtypes (pDCs, CD8(+) cDCs and CD8(-) cDCs). We also isolated an in vivo equivalent of the DC-committed pro-DC precursor cell, which also gave rise to the three DC subtypes. Clonal analysis of the progeny of individual pro DC precursors demonstrated that some pro-DC precursors gave rise to all three DC subtypes, some produced cDCs but not pDCs, and some were fully committed to a single DC subtype. Thus, commitment to particular DC subtypes begins mainly at this pro-DC stage. PMID- 17922016 TI - Identification of clonogenic common Flt3+M-CSFR+ plasmacytoid and conventional dendritic cell progenitors in mouse bone marrow. AB - Lymphoid tissue plasmacytoid and conventional dendritic cells (DCs) are continuously regenerated from hematopoietic stem cells. The cytokine dependence and biology of plasmacytoid and conventional DCs suggest that regeneration might proceed through common DC-restricted developmental intermediates. By selecting for cytokine receptor expression relevant to DC development, we identify here highly cycling Lin(-)c-Kit(int)Flt3(+)M-CSFR(+) cells with a distinct gene expression profile in mouse bone marrow that, on a clonal level in vitro and as a population both in vitro and in vivo, efficiently generated plasmacytoid and conventional DCs but no other lineages, which increased in number after in vivo injection of the cytokine Flt3 ligand. These clonogenic common DC progenitors thus define a cytokine-regulated DC developmental pathway that ensures the supply of various DC populations. PMID- 17922017 TI - In vivo gene regulation in Salmonella spp. by a salicylate-dependent control circuit. AB - Systems allowing tightly regulated expression of prokaryotic genes in vivo are important for performing functional studies of bacterial genes in host-pathogen interactions and establishing bacteria-based therapies. We integrated a regulatory control circuit activated by acetyl salicylic acid (ASA) in attenuated Salmonella enterica that carries an expression module with a gene of interest under control of the XylS2-dependent Pm promoter. This resulted in 20-150-fold induction ex vivo. The regulatory circuit was also efficiently induced by ASA when the bacteria resided in eukaryotic cells, both in vitro and in vivo. To validate the circuit, we administered Salmonella spp., carrying an expression module encoding the 5-fluorocytosine-converting enzyme cytosine deaminase in the bacterial chromosome or in a plasmid, to mice with tumors. Induction with ASA before 5-fluorocytosine administration resulted in a significant reduction of tumor growth. These results demonstrate the usefulness of the regulatory control circuit to selectively switch on gene expression during bacterial infection. PMID- 17922018 TI - Comprehensive analysis of diverse ribonucleoprotein complexes. AB - The study of the dynamic interactome of cellular ribonucleoprotein (RNP) particles has been hampered by severe methodological limitations. In particular, the affinity purification of intact RNP complexes from cell lysates suffers from RNA degradation, loss of interacting macromolecules and poor overall yields. Here we describe a rapid affinity-purification method for efficient isolation of the subcomplexes that dynamically organize different RNP biogenesis pathways in Saccharomyces cerevisiae. Our method overcomes many of the previous limitations to produce large RNP interactomes with almost no contamination. PMID- 17922019 TI - The role of the complement cascade in endotoxin-induced septic encephalopathy. AB - The complement system normally eliminates bacteria and has a protective effect. However, in an inflammatory setting such as sepsis, an exaggerated or insufficient activation of this cascade can have deleterious effect through the activation of glial cells, secretion of proinflammatory cytokines and generation of other toxic products. The aim of the present study was to investigate the role of the complement cascade in septic encephalopathy, through the passive injection of endotoxin/lipopolysaccharide (LPS) into mice overexpressing the potent complement inhibitor, CR1-related y (Crry-tg). Increased gliosis occurred in brains of endotoxemic mice. Concomitant with this, there was a significant rise in mRNA expression of GFAP, CD45 and proinflammatory molecules, TLR4, TNF-alpha and NO, in these brains. Consistent with the capacity of these inflammatory mediators, there was increased apoptosis as determined by DNA fragmentation and TUNEL staining on LPS treatment, which occurred through the Akt pathway. In addition, there was increased water content in brain, similar to cerebral edema observed in sepsis. Relative to wild-type mice, complement-inhibited mice had an attenuated inflammatory response, decreased edema and reduced apoptosis. Therefore, we demonstrate for the first time that the complement cascade appears to be one of the key players that cause brain pathology in an endotoxemic setting and therefore is a viable therapeutic target. PMID- 17922021 TI - PTEN: a promising pharmacological target to enhance epithelial wound healing. AB - PI3Ks (phosphoinositide-3 kinases) produce PIP3 (phosphatidylinositol(3,4,5) trisphosphate) which mediates signals for cell survival and proliferation. The tumour suppressor PTEN (phosphatase and tensin homologue) dephosphorylates PIP3 and is a key negative regulator of PI3K signalling. Recent research highlighted important roles for PI3K/PTEN in cell polarization and directional cell migration, pointing to a significant role for PTEN in wound healing where spatially organized tissue growth is essential. Lai et al. (in this issue of British Journal of Pharmacology) have moved a step closer in utilizing PTEN for wound healing through pharmacological inhibition. Two vanadium derivative inhibitors targeting PTEN significantly elevated the level of phosphorylated Akt (protein kinase B) and nearly doubled the wound healing rate in monolayer cultures of lung and airway epithelial cells. Damage to airway and lung epithelia underlies a wide spectrum of significant clinical conditions. With further experiments, this promising approach may find potential clinical use in situations where enhanced wound healing of pulmonary and other epithelia is important. PMID- 17922020 TI - EMT tumorigenesis in the mouse mammary gland. AB - The term EMT (epithelial-mesenchymal transition) is used in many settings. This term is used to describe the mechanisms facilitating cellular repositioning and redeployment during embryonic development and tissue reconstruction after injury. Recently, EMT has also been applied to potential mechanisms for malignant progression and has appeared as a specific diagnostic category of tumors. In mice, most 'EMT' tumors have a spindle cell phenotype. The definition of EMT is controversial because spindle cell tumors are not common in humans, especially in human breast cancers. Spindle cell tumors of the mouse mammary gland have been observed for many years where they are usually classified as sarcomas or carcinosarcomas. Genetically engineered mice develop mammary spindle cell tumors that appear to arise in the epithelium and undergo EMT. To better understand the origin and evolution of these spindle cell tumors in progression and metastases, seven cohorts of spindle cell tumors from the archives of the University of California, Davis Mutant Mouse Pathology Laboratory were studied. This study provides experimental and immunohistochemical evidence of EMT showing that dual epithelial and mesenchymal staining of tumor spindle cells identifies some, but not all, EMT-type tumors in the mouse. This suggests that potential EMT tumors are best designated EMT-phenotype tumors. PMID- 17922022 TI - Phosphatase and tensin homologue deleted on chromosome ten (PTEN) as a molecular target in lung epithelial wound repair. AB - BACKGROUND AND PURPOSE: Epithelial injury contributes to lung pathogenesis. Our work and that of others have identified the phosphoinositide-3 kinase (PI3K)/Akt pathway as a vital component of survival in lung epithelia. Therefore, we hypothesized that pharmacological inhibition of PTEN, a major suppressor of this pathway, would enhance wound closure and restore lung epithelial monolayer integrity following injury. EXPERIMENTAL APPROACH: We evaluated the ability of two bisperoxovanadium derivatives, bpV(phen) and bpV(pic), in differentiated primary human airway epithelia and BEAS2B cultures for their ability to inhibit PTEN, activate the PI3K/Akt pathway and restore epithelial monolayer integrity following mechanical injury. KEY RESULTS: BpV(phen) and bpV(pic) induced Akt phosphorylation in primary and BEAS2B cells in a dose and time dependent manner. Minimal toxicity was observed as measured by lactate dehydrogenase (LDH) release. To verify that Akt phosphorylation is specifically induced by PTEN inhibition, the PTEN positive cell line, DU145, and two PTEN negative cell lines, LNCaP and PC3, were examined. PTEN positive cells demonstrated a dose responsive increase in Akt phosphorylation whereas PTEN negative cells showed no response indicating that bpV(phen) directly suppresses PTEN without affecting auxiliary pathways. Next, we observed that exposure to either compound resulted in accelerated wound closure following mechanical injury. Similar effects were observed after transfection with a dominant negative isoform of PTEN and PTEN specific siRNA. CONCLUSIONS AND IMPLICATIONS: From these studies, we conclude that PTEN is a valid target for future studies directed at restoring epithelial barrier function after lung injury. PMID- 17922023 TI - Flow-induced enhancement of vasoconstriction and blockade of endothelium-derived hyperpolarizing factor (EDHF) by ascorbate in the rat mesentery. AB - BACKGROUND AND PURPOSE: We previously reported that ascorbate inhibits flow- and agonist-induced, EDHF-mediated vasodilatation in the bovine ciliary circulation. This study examined whether ascorbate had similar actions in the rat mesenteric vasculature. EXPERIMENTAL APPROACH: The effects of ascorbate were examined both in rat second order mesenteric arterial rings suspended in a static wire myograph and the rat mesentery perfused at different rates of flow. KEY RESULTS: Ascorbate (50 microM) had no effect on U46619-induced tone or acetylcholine-induced, EDHF mediated vasodilatation in either rings of mesenteric artery or the perfused mesentery at rates of flow below 10 ml min(-1). At higher rates of flow, ascorbate produced two distinct effects in the rat mesentery: a rapid and maintained enhancement of vasoconstrictor tone and a slow (max at 3 h) inhibition of acetylcholine-induced, EDHF-mediated vasodilatation. The enhancement of vasoconstrictor tone appeared to be due to inhibition of flow-induced EDHF-like activity, since it was endothelium-dependent, but could be elicited during blockade of nitric oxide synthase and cyclooxygenase. Despite this, the classical inhibitors of EDHF, apamin and charybdotoxin, failed to affect the ascorbate induced enhancement of tone, although they inhibited acetylcholine-induced vasodilatation. CONCLUSIONS AND IMPLICATIONS: Ascorbate inhibits both flow- and agonist-induced EDHF in the rat mesentery. The strikingly different timecourses of these two effects, together with their differential sensitivity to apamin and charybdotoxin, suggest that the flow- and agonist-induced EDHFs in the rat mesenteric vasculature may either be different entities or operate by different mechanisms. PMID- 17922024 TI - Agonist-dependent cannabinoid receptor signalling in human trabecular meshwork cells. AB - BACKGROUND AND PURPOSE: Trabecular meshwork (TM) is an ocular tissue involved in the regulation of aqueous humour outflow and intraocular pressure (IOP). CB1 receptors (CB1) are present in TM and cannabinoid administration decreases IOP. CB1 signalling was investigated in a cell line derived from human TM (hTM). EXPERIMENTAL APPROACH: CB1 signalling was investigated using ratiometric Ca2+ imaging, western blotting and infrared In-Cell Western analysis. KEY RESULTS: WIN55212-2, a synthetic aminoalkylindole cannabinoid receptor agonist (10-100 microM) increased intracellular Ca2+ in hTM cells. WIN55,212-2-mediated Ca2+ increases were blocked by AM251, a CB1 antagonist, but were unaffected by the CB2 antagonist, AM630. The WIN55,212-2-mediated increase in [Ca2+]i was pertussis toxin (PTX)-insensitive, therefore, independent of Gi/o coupling, but was attenuated by a dominant negative Galpha(q/11) subunit, implicating a Gq/11 signalling pathway. The increase in [Ca2+]i was dependent upon PLC activation and mobilization of intracellular Ca2+ stores. A PTX-sensitive increase in extracellular signal-regulated kinase (ERK1/2) phosphorylation was also observed in response to WIN55,212-2, indicative of a Gi/o signalling pathway. CB1-Gq/11 coupling to activate PLC-dependent increases in Ca2+ appeared to be specific to WIN55,212-2 and were not observed with other CB1 agonists, including CP55,940 and methanandamide. CP55940 produced PTX-sensitive increases in [Ca2+]i at concentrations>or=15 microM, and PTX-sensitive increases in ERK1/2 phosphorylation. CONCLUSIONS AND IMPLICATIONS: This study demonstrates that endogenous CB1 couples to both Gq/11 and Gi/o in hTM cells in an agonist dependent manner. Cannabinoid activation of multiple CB1 signalling pathways in TM tissue could lead to differential changes in aqueous humour outflow and IOP. PMID- 17922025 TI - Direct suppression of autoreactive lymphocytes in the central nervous system via the CB2 receptor. AB - The cannabinoid system is now recognized as a regulator of both the nervous and immune systems. Although marijuana has been used for centuries for the treatment of a variety of disorders, its therapeutic mechanisms are only now being understood. The best-studied plant cannabinoid, delta9-tetrahydrocannabinol (THC), produced by Cannabis sativa and found in marijuana, has shown evidence of being immunosuppressive in both in vivo and in vitro. Since THC binds to at least two receptors that are differentially expressed by the immune and nervous systems, it has not been possible to clearly discriminate the biological effects it exerts in the two systems. In addition, endogenous cannabinoids have also been described that bind to both receptors and exert both neuronal and immune modulatory activity. The generation of mice deficient in specific cannabinoid receptors has facilitated studies to discriminate cannabinoid-specific functions. This review focuses on the function of the cannabinoid receptor 2 (CB2), primarily expressed in the immune system, in regulating T cell effector functions associated with autoimmune inflammation in the central nervous system (CNS). PMID- 17922026 TI - Angiotensin AT1 receptor antagonists exert anti-inflammatory effects in spontaneously hypertensive rats. AB - BACKGROUND AND PURPOSE: Although the main therapeutic effect of angiotensin AT1 receptor antagonists is to decrease blood pressure, they also exert anti inflammatory effects in the cardiovascular system. However, the underlying mechanisms remain unclear. We investigated the inhibitory effect of AT1 antagonists on the chemokine monocyte chemoattractant protein 1 (MCP-1) and its receptor C-C chemokine receptor 2 (CCR2) in rat monocytes and aortas. EXPERIMENTAL APPROACH: Spontaneous hypertensive rats (SHRs) were treated with the AT1 antagonists losartan or telmisartan for 4 weeks, and Wistar-Kyoto rats (WKYs) were used as normotensive controls. Systolic arterial pressure was measured, and the number of macrophages in the aortic vessel wall was assessed by anti-ED-1 antibody immunolabelling. KEY RESULTS: Compared with WKYs, SHRs showed significantly increased ED-1 positive macrophages in the aortic wall, which were decreased after high doses of losartan or telmisartan. Low doses of losartan did not improve blood pressure significantly as did the high doses, but markedly decreased macrophage infiltration in the vessel wall. AT1 antagonists, particularly at high doses, improved aortic remodeling in SHR. At the molecular level, AT1 antagonists attenuated the expression of MCP-1 and CCR2 in the aorta and peripheral blood monocytes and lowered the serum level of MCP-1. In addition, Western blotting showed that AT1 antagonists inhibited the phosphorylation of Akt in mouse monocytes. CONCLUSIONS AND IMPLICATIONS: AT1 antagonism inhibited vessel wall inflammation and inhibition of PI3K/Akt may be involved in the modulation of the MCP-1/CCR2 system by AT1 antagonists in SHRs. PMID- 17922027 TI - The requirement for SNF5/INI1 in adipocyte differentiation highlights new features of malignant rhabdoid tumors. AB - ATP-dependent SWI/SNF chromatin remodeling complexes regulate cell-cycle and play critical roles in a variety of differentiation pathways. The core subunit SNF5/INI1 is a tumor suppressor that is inactivated in a highly aggressive childhood cancer of unknown cellular origin, termed malignant rhabdoid tumor (MRT). The highly undifferentiated phenotype of this tumor suggests that the loss of-function of hSNF5/INI1 impairs specific differentiation programs of the MRT parental cell. Based on the hypothesis that these programs might be reinitialized upon hSNF5/INI1 re-expression in MRTs, we show that some MRT cell lines can differentiate toward the adipogenic lineage. We further show that the knock down of the SNF5/INI1 subunit abrogates adipocyte differentiation of murine 3T3-L1 preadipocytes and of human mesenchymal stem cells. Finally, we provide evidence that hSNF5/INI1 cooperates with C/EBPbeta and PPARgamma2 transcriptional regulators to activate the expression of adipocyte-specific genes. These data indicate that not only the ATPase subunit of the SWI/SNF complex, but also SNF5/INI1 is required for adipocyte differentiation. They further show that MRT cell lines harbor an adipogenic differentiation potential and that the tumor suppressor role of the SNF5/INI1 subunit may rely on its ability to regulate the balance between cell proliferation and differentiation. PMID- 17922028 TI - Ras mediates cell survival by regulating tuberin. AB - Mutational activation of Ras promotes oncogenesis by controlling cell cycle regulation and cell survival. Ras-mediated activation of both, the PI3K/AKT pathway and the MEK/ERK pathway, can trigger downregulation of the function of tuberin to block the activities of mTOR and p70S6K. Here we demonstrate that Ras induced cell survival is accompanied by upregulation of p70S6K activity. Ras harbors the potential to negatively affect tuberin-induced apoptosis and p70S6K inactivation. These effects of Ras were found to depend on its potential to regulate the MEK/ERK pathway. Experiments using tuberin-negative fibroblasts revealed that the potential of Ras to counteract apoptosis depends on functional tuberin. Taken together, we provide evidence that the function of Ras to trigger inactivation of tuberin plays a major role in the regulation of cell survival upon mutational activation of the oncogene Ras. This is the first description of a functional interaction between the tumor suppressor tuberin and the oncogene Ras in regulating apoptosis. PMID- 17922029 TI - Complex patterns of ETS gene alteration arise during cancer development in the human prostate. AB - An ERG gene 'break-apart' fluorescence in situ hybridization (FISH) assay has been used to screen whole-mount prostatectomy specimens for rearrangements at the ERG locus. In cancers containing ERG alterations the observed pattern of changes was often complex. Different categories of ERG gene alteration were found either together in a single cancerous region or within separate foci of cancer in the same prostate slice. In some cases the juxtaposition of particular patterns of ERG alterations suggested possible mechanisms of tumour progression. Prostates harbouring ERG alterations commonly also contained cancer that lacked rearrangements of the ERG gene. A single trans-urethral resection of the prostate specimen examined harboured both ERG and ETV1 gene rearrangements demonstrating that the observed complexity may, at least in part, be explained by multiple ETS gene alterations arising independently in a single prostate. In a search for possible precursor lesions clonal ERG rearrangements were found both in high grade prostatic intraepithelial neoplasia (PIN) and in atypical in situ epithelial lesions consistent with the diagnosis of low grade PIN. Our observations support the view that ERG gene alterations represent an initiating event that promotes clonal expansion initially to form regions of epithelial atypia. The complex patterns of ERG alteration found in prostatectomy specimens have important implications for the design of experiments investigating the clinical significance and mechanism of development of individual prostate cancers. PMID- 17922030 TI - The N-methyl-D-aspartate receptor type 2A is frequently methylated in human colorectal carcinoma and suppresses cell growth. AB - N-methyl-D-aspartate receptors (NMDARs) are the predominant excitatory neurotransmitter receptors in the mammalian brain. We found that among the three NMDARs examined (NMDAR1, NMDAR2A, NMDAR2B), only NMDAR2A was silenced in colorectal carcinoma (CRC) cell lines at basal line and reactivated by the demethylating agent, 5-aza-2'-deoxycytidine. NMDAR2A was expressed in normal colon epithelium, while expression was hardly detectable in colon cancer tissues. Promoter methylation of NMDAR2A was confirmed by bisulfite sequencing and combined bisulfite restriction analysis in the CRC cell lines and primary tumors. Quantitative methylation-specific PCR demonstrated NMDAR2A promoter hypermethylation in 82 of 100 primary human CRC, 15 of 100 normal corresponding epithelial tissues and 1 of 11 (9%) normal colon mucosa samples obtained from patients without cancer. Moreover, forced expression of full-length NMDAR2A in CRC cell lines induced apoptosis and almost abolished the ability of the cells to form colonies in culture, while NMDAR2A knockdown increased cell growth. Thus, NMDAR2A is commonly hypermethylated in primary human CRC and possesses tumor suppressive activity. PMID- 17922031 TI - The RhoGEF domain of p210 Bcr-Abl activates RhoA and is required for transformation. AB - The BCR-ABL oncogene encodes an in-frame fusion protein containing N-terminal sequences derived from Bcr and C-terminal sequences derived from Abl. Bcr contains a centrally located Rho-specific guanine nucleotide exchange factor (RhoGEF) domain that is retained within p210 Bcr-Abl. Although this domain is subject to autoinhibition in the context of Bcr, here we show that it is constitutively activated in p210 Bcr-Abl. p210 Bcr-Abl can stimulate RhoA activation independently of its tyrosine kinase activity, and mutations within the RhoGEF domain that are predicted to eliminate RhoGEF activity inhibit RhoA activation. The RhoGEF mutant of p210 Bcr-Abl does not affect the tyrosine kinase activity of the molecule, nor the ability of p210 Bcr-Abl to interact with XPB through the RhoGEF domain. Despite retaining normal levels of tyrosine kinase activity, the RhoGEF mutant of p210 Bcr-Abl is impaired in transforming activity as measured by anchorage-independent growth. However, the mutant is still able to confer the phenotype of growth factor independence in myeloid cells, suggesting that some, but not all parameters of p210 Bcr-Abl transformation, are dependent upon a catalytically active RhoGEF domain. Collectively, these observations identify a gain-of-function activity attributable to the RhoGEF domain of p210 Bcr-Abl that is required to support the transformed phenotype. PMID- 17922033 TI - MicroRNA expression signature of human sarcomas. AB - MicroRNAs (miRNAs) are approximately 22 nucleotide-long noncoding RNAs involved in several biological processes including development, differentiation and proliferation. Recent studies suggest that knowledge of miRNA expression patterns in cancer may have substantial value for diagnostic and prognostic determinations as well as for eventual therapeutic intervention. We performed comprehensive analysis of miRNA expression profiles of 27 sarcomas, 5 normal smooth muscle and 2 normal skeletal muscle tissues using microarray technology and/or small RNA cloning approaches. The miRNA expression profiles are distinct among the tumor types as demonstrated by an unsupervised hierarchical clustering, and unique miRNA expression signatures were identified in each tumor class. Remarkably, the miRNA expression patterns suggested that two of the sarcomas had been misdiagnosed and this was confirmed by reevaluation of the tumors using histopathologic and molecular analyses. Using the cloning approach, we also identified 31 novel miRNAs or other small RNA effectors in the sarcomas and normal skeletal muscle tissues examined. Our data show that different histological types of sarcoma have distinct miRNA expression patterns, reflecting the apparent lineage and differentiation status of the tumors. The identification of unique miRNA signatures in each tumor type may indicate their role in tumorigenesis and may aid in diagnosis of soft tissue sarcomas. PMID- 17922032 TI - MTA1-mediated transcriptional repression of BRCA1 tumor suppressor gene. AB - Metastasis-associated tumor antigen 1 (MTA1), a component of the nucleosome remodeling and deacetylating (NuRD) complex is routinely upregulated in several cancers. In the present study, we investigated the potential role of MTA1 in BRCA1 transcriptional repression and subsequent chromosomal instability. MTA1 NuRD complex was found to negatively regulate BRCA1 transcription by physically associating with an atypical estrogen-responsive element (ERE) on the BRCA1 promoter. Moreover, MTA1 and HDAC complex recruited to the ERE of BRCA1 promoter in an ER alpha-dependent manner. Accordingly, BRCA1 protein levels were enhanced by silencing of either MTA1 expression or by treatment with the specific histone deacetylase inhibitor trichostatin A. MTA1's strong repressive effects on BRCA1 expression was supported by our observation that cells stably overexpressing MTA1 showed centrosome amplification which has been long implicated as a phenotype for BRCA1 repression. Accordingly, overexpression of BRCA1 in cells stably over expressing MTA1 resulted in restoration of normal centrosome numbers. Together, these findings strongly implicate MTA1 in the transcriptional repression of BRCA1 leading to abnormal centrosome number and chromosomal instability. PMID- 17922034 TI - PAX-FKHR function as pangenes by simultaneously inducing and inhibiting myogenesis. AB - Alveolar rhabdomyosarcomas (ARMS) escape terminal differentiation despite exhibiting a skeletal muscle phenotype. To understand the role of the ARMS specific PAX-FKHR proteins in myogenesis, we characterized their regulation of MyoD expression and function. Reporter assays show that PAX-FKHR transactivate MyoD expression through its 258 bp core enhancer. Gel-shift assays confirm that PAX-FKHR bind to core enhancer sequences showing similarity to consensus PAX3/PAX FKHR-binding sites. We show that while PAX3-FKHR activates the expression of endogenous MyoD and myogenin proteins in transduced NIH3T3 fibroblasts, it inhibits them from terminally differentiating as shown by low myogenin and myosin heavy chain expression, and lack of myotube formation. Attenuation of MyoD transcriptional activity via phosphorylation coupled to the lack of cell cycle arrest is the underlying mechanism for the differentiation block. Lastly, we show that fibroblast growth factor receptor signaling likely mediates the inhibition of differentiation by PAX3-FKHR. In a single experimental system we demonstrate that PAX3-FKHR can simultaneously induce myogenesis while preventing its completion. We propose a model whereby PAX-FKHR commit a yet undefined precursor cell to the myogenic lineage while at the same time inhibit terminal differentiation, thereby contributing to ARMS formation. PMID- 17922035 TI - Heat shock factor 1 represses estrogen-dependent transcription through association with MTA1. AB - Heat shock factor 1 (HSF1), the transcriptional activator of the heat shock genes, is increasingly implicated in cancer. We have shown that HSF1 binds to the corepressor metastasis-associated protein 1 (MTA1) in vitro and in human breast carcinoma samples. HSF1-MTA1 complex formation was strongly induced by the transforming ligand heregulin and complexes incorporated a number of additional proteins including histone deacetylases (HDAC1 and 2) and Mi2alpha, all components of the NuRD corepressor complex. These complexes were induced to assemble on the chromatin of MCF7 breast carcinoma cells and associated with the promoters of estrogen-responsive genes. Such HSF1 complexes participate in repression of estrogen-dependent transcription in breast carcinoma cells treated with heregulin and this effect was inhibited by MTA1 knockdown. Repression of estrogen-dependent transcription may contribute to the role of HSF1 in cancer. PMID- 17922037 TI - High-dose chemotherapy with peripheral blood progenitor cell support for patients with non-small cell lung cancer: the experience of the European Group for Bone Marrow Transplantation (EBMT) Solid Tumours Working Party. AB - We report the experience of the EBMT Solid Tumours Working Party (STWP) using high-dose chemotherapy (HDCT) with PBPC support in patients with non-small cell lung cancer (NSCLC). Between 1989 and 2004, 36 NSCLC patients (27 men and 9 women), median age 53.5 years (range: 24-62) were treated with 63 HDCT courses. A high-dose carboplatin-based regimen was used in 53% of the cases. Thirty-two patients had relapsed/metastatic disease, while four classified as stage IIIB received HDCT followed by radiotherapy. No treatment-related death occurred. Of 25 patients who were planned to receive multi-cycle HDCT, 4 cases (16%) interrupted the treatment early due to prolonged severe toxicities and 4 (16%) due to progressive disease. Of 36 evaluable patients, 3 (8%) achieved a complete remission and 13 (36%) had a partial remission at an overall response rate of 44%. Of these, one patient with stage IIIB and one with stage IV are alive disease free at 71+ and 149+ months, respectively. After a median follow-up of 48 months (range: 6-149), median survival was 7 months (range: 1-149). Despite one anecdotal case, HDCT did not show significant activity, but induced relevant morbidity in NSCLC patients. PMID- 17922039 TI - Growth and development after hematopoietic cell transplant in children. AB - Hematopoietic cell transplantation (HCT) following high-dose chemotherapy or chemoradiotherapy for children with malignant or nonmalignant hematologic disorders has resulted in an increasing number of long-term disease-free survivors. The preparative regimens include high doses of alkylating agents, such as CY with or without BU, and may include TBI. These agents impact the neuroendocrine system in growing children and their subsequent growth and development. Children receiving high-dose CY or BUCY have normal thyroid function, but those who receive TBI-containing regimens may develop thyroid function abnormalities. Growth is not impacted by chemotherapy-only preparative regimens, but TBI is likely to result in growth hormone deficiency and decreased growth rates that need to be treated with synthetic growth hormone therapy. Children who receive high-dose CY-only have normal development through puberty, whereas those who receive BUCY have a high incidence of delayed pubertal development. Following fractionated TBI preparative regimens, approximately half of the patients have normal pubertal development. These data demonstrate that the growth and development problems after HCT are dependent upon the preparative regimen received. All children should be followed for years after HCT for detection of growth and development abnormalities that are treatable with appropriate hormone therapy. PMID- 17922036 TI - The inhibitory effects of Disabled-2 (Dab2) on Wnt signaling are mediated through Axin. AB - beta-Catenin-mediated Wnt signaling is essential in embryonic development and in adult tissues. Recent studies have demonstrated that Axin not only plays an important inhibitory role in coordinating beta-catenin degradation, but is itself degraded by the low-density-lipoprotein receptor-related protein (LRP)5/6 Wnt co receptor. Here, we demonstrate that the endocytic adaptor molecule Disabled-2 (Dab2), which we have previously demonstrated to act as an inhibitor of beta catenin signaling, interacts with Axin and prevents its interaction with and degradation by the LRP5 co-receptor, thereby increasing its half-life and stabilization. Dab2 levels induced during retinoic acid-induced differentiation of F9, or during transforming growth factor-beta-induced epithelial-mesenchymal transdifferentiation of mouse mammary epithelial cells result in the stabilization of Axin and concomitant inhibition of beta-catenin signaling. Ectopic expression of Dab2 in F9 cells as well as in transformed cell lines results in increased Axin expression and attenuation of Wnt-mediated signaling. We conclude that Dab2 may play an important role in the maintenance of the differentiated state and restrain Wnt-mediated proliferation through its association with and modulation of Axin. PMID- 17922040 TI - Central nervous system graft-versus-host disease: consider progressive multifocal leukoencephalopathy among the differential diagnoses. PMID- 17922041 TI - Anti-leukaemic role of acute GvHD after unrelated haematopoietic stem cell transplantation in intermediate- to high-risk acute myelogenous leukaemia. AB - Little is known about the role of acute GvHD (aGvHD) based on the concept of graft-versus-leukaemia effect (GVLE) after unrelated donor haematopoietic stem cell transplantation (uHSCT). We evaluated 67 uHSCTs performed with multinational unrelated donors for patients with AML. The median follow-up duration was 18 months (range 7-61). The majority of patients had intermediate- or high-risk cytogenetic findings. The conditioning regimen for most patients consisted of cyclophosphamide plus total body irradiation (n=56) with our standard GvHD prophylaxis containing tacrolimus plus a short course of methotrexate. The incidence of aGvHD and chronic GvHD was 50 and 52%, respectively. Eight patients (12%) have relapsed to date. The estimated overall disease-free survival (DFS) rate at 5 years was 67%. Notably fewer relapses were seen when aGvHD developed (P=0.008). Specifically, high-risk AML patients had a much lower relapse rate when they developed aGvHD (P=0.01), compared with the intermediate-risk group. Therefore, the development of aGvHD after uHSCT in AML patients is closely related to a lower relapse rate, probably in association with GVLE. PMID- 17922042 TI - Current status and future perspectives for yttrium-90 ((90)Y)-ibritumomab tiuxetan in stem cell transplantation for non-Hodgkin's lymphoma. AB - Haematopoietic SCT is currently considered a therapeutic option mainly in relapsed or refractory non-Hodgkin's lymphoma (NHL) owing to high post transplantation relapse rates and significant toxicity of conventional myeloablative conditioning for allogeneic SCT. Radiolabelled immunotherapy combines the benefits of monoclonal antibody targeting with therapeutic doses of radiation, and is a promising advance in the treatment of malignant lymphomas. It is now under investigation as a component of conditioning prior to SCT, with the aim of improving outcomes following SCT without increasing the toxicity of high dose chemotherapy pre-transplant conditioning. An expert panel met at a European workshop in November 2006 to review the latest data on radiolabelled immunotherapy in the transplant setting, and its potential future directions, with a focus on (90)Y-ibritumomab tiuxetan. They reviewed data on the combination of standard/high/escalating dose (90)Y-ibritumomab tiuxetan with high-dose chemotherapy, and high/escalating dose (90)Y-ibritumomab tiuxetan as the sole myeloablative agent, prior to autologous SCT, and also (90)Y-ibritumomab tiuxetan as a component of reduced intensity conditioning prior to allogeneic SCT. The preliminary data are highly promising in terms of conditioning tolerability and patient outcomes following transplant; further phase II studies are now needed to consolidate these data and to investigate specific patient populations and NHL subtypes. PMID- 17922043 TI - The impact of histologic grade on the outcome of high-dose therapy and autologous stem cell transplantation for follicular lymphoma. AB - The impact of the follicular lymphoma (FL) histologic grade on outcomes after high-dose therapy (HDT) and autologous stem cell transplantation (ASCT) is unknown. We evaluated 219 consecutive patients with grades 1-3 FL who underwent HDT and ASCT at our center. Overall survival (OS), progression-free survival (PFS), relapse and non-relapse mortality (NRM) was estimated for each grade after controlling for other predictive factors. The number of patients with grades 1, 2 and 3 FL was 106 (48%), 75 (34%) and 38 (17%), respectively. Five-year outcome estimates for the entire cohort included 60% OS, 39% PFS and 46% relapse (median follow-up=7.8 years). PFS and relapse were nearly identical among patients with grade 3 FL versus grades 1-2 FL after adjusting for other contributing factors (hazard ratio (HR)=0.90, P=0.68; HR=1.07, P=0.80, respectively). The hazard for mortality (HR=0.70, P=0.23) and NRM (HR=0.33, P=0.07) was non-significantly lower among patients with grade 3 FL compared to patients with grades 1-2 disease. Factors associated with inferior PFS included elevated lactate dehydrogenase (HR=1.52, P=0.03), chemoresistance (HR=1.82, P=0.02), > or =2 prior therapies (HR=1.8, P=0.03) and prior radiation (HR=1.99, P=0.003). These data suggest that the histologic grade of FL does not impact PFS or relapse following HDT and ASCT. PMID- 17922044 TI - Type VII secretion--mycobacteria show the way. AB - Recent evidence shows that mycobacteria have developed novel and specialized secretion systems for the transport of extracellular proteins across their hydrophobic, and highly impermeable, cell wall. Strikingly, mycobacterial genomes encode up to five of these transport systems. Two of these systems, ESX-1 and ESX 5, are involved in virulence - they both affect the cell-to-cell migration of pathogenic mycobacteria. Here, we discuss this novel secretion pathway and consider variants that are present in various Gram-positive bacteria. Given the unique composition of this secretion system, and its general importance, we propose that, in line with the accepted nomenclature, it should be called type VII secretion. PMID- 17922045 TI - Chemosensory pathways, motility and development in Myxococcus xanthus. AB - The complex life cycle of Myxococcus xanthus includes predation, swarming, fruiting-body formation and sporulation. The genome of M. xanthus is large and comprises an estimated 7,400 open reading frames, of which approximately 605 code for regulatory genes. These include eight clusters of chemotaxis-like genes that define eight chemosensory pathways, most of which have dedicated functions. Although many of these chemosensory pathways have a role in controlling motility, at least two of these pathways control gene expression during development. PMID- 17922046 TI - HER-2, TOP2A and chromosome 17 alterations in breast cancer. AB - HER-2 amplification is a biomarker for identifying patients who respond to trastuzumab and has been evaluated as a factor predicting the response to anthracyclines. The relationship between HER-2 and response to anthracycline therapy may also be the result of the close localization of TOP2A on 17q. It has been a matter of debate whether these two genes, HER-2 and TOP2A, behave separately on different amplicons or act together thus making it possible to predict the TOP2A status from the HER-2 status. In this study TOP2A, HER-2 and chromosome 17 aneusomy were investigated by fluorescent in situ hybridization (FISH) in 50 consecutive breast cancer patients. HER-2 amplification was detected in 11 patients (22%) and TOP2A changes were seen in 6 patients (12%); two amplifications and two deletions were observed in HER-2-amplified cases and two deletions in HER-2-nonamplified cases. Three of the TOP2A-deleted cases had polysomy 17. HER-2 copy number was higher than the TOP2A copy number in one patient with co-amplification. Polysomy was observed in 9 cases (18%) and monosomy in 6 cases (12%). Aneusomy was the sole anomaly in 11 patients (22%). We conclude that the TOP2A status cannot be predicted from the HER-2 status and evaluation of the TOP2A status only in patients with HER-2 overexpression may lead to missing cases with TOP2A deletion with possible resistance to therapy. Other factors modulating topo II activity may also affect the response to therapy. Studies evaluating different parameters that can modulate topo II activity and the response to the drugs targeting the enzyme are necessary. PMID- 17922047 TI - Detection of bladder cancer from the urine using fluorescence in situ hybridization technique. AB - The authors report on their first experiences with the UroVysion fluorescence in situ hybridization (FISH) kit developed for the detection of bladder cancer. This new non-invasive diagnostic application of the FISH technique in the field of urology was elaborated to replace cystoscopy. The special urine examination method detects genetic alterations of the urothelial cells found in the urine, using fluorescent directlabeled DNA probes binding to the peri-centromeric regions of chromosomes 3, 7 and 17 as well as on the 9p21 locus. We aimed to evaluate the utility of UroVysion test in the light of the histological diagnosis. Urine samples from 43 bladder cancer patients and 12 patients with no or benign alterations were studied using a new application of FISH technique: the UroVysion reagent kit. The obtained FISH results were compared with the histological findings of the transurethral surgical resection specimens. The study rated the specificity and sensitivity of the technique 100% and 87%, respectively. Therefore, the technique could well fit into the diagnostic process of bladder carcinomas. Statistical analyses showed significant correlation between tumor progression and the severity of the genetic alterations detected by this FISH technique. Furthermore, positive correlation was found between tumor grade and the proportion of tumor cells showing genetic abnormality. The noninvasiveness, the robustness of evaluation and the high specificity/sensitivity are all in favor of this technique. The disadvantages are the higher costs of the technical background and the required future clinical studies to determine whether this technique can replace cystoscopy. PMID- 17922048 TI - Gender-related hormonal risk factors for oral cancer. AB - Oral cancer (OC) is a neoplasm with fairly high male to female ratio in most populations. The conspicuously lower incidence of this tumor among women than man is suggestive of certain endocrine involvement in its development. The aim of the present case-control study was to clarify the origin of this gender-specific risk of OC incidence. 2660 inpatients (530 females and 2130 males) with squamous cell OC at the Department of Oral and Maxillofacial Surgery were included in a case control study. Smoking, alcohol consumption, elevated fasting serum glucose level and menopausal histories of female cases were registered. Smoking and excessive alcohol intake proved to be strong risk factors for OC both in the male and female group. However, moderate alcohol consumption was a weaker risk factor for male patients, and it presented no risk for female cases. Elevated fasting glucose level was not a demonstrable OC risk factor among males, however, it proved to be strong risk factor for OC among female patients, especially in gingival cancer cases. The almost exclusively postmenopausal state of female OC patients and the long mean interval (17 years) between their menopause and OC diagnosis suggested an important role of estrogen deficiency in OC epidemiology. The significantly younger mean age at menopause and the significantly higher rate of hysterectomy among female OC cases in comparison with their controls also support the estrogen deficiency hypothesis. This novel hypothesis of estrogen deficiency and elevated fasting glucose as risk factors for OC in postmenopausal women may provide new insights into the etiology of oral malignancies. PMID- 17922049 TI - Expression of P-glycoprotein and metallothionein in gastrointestinal stromal tumor and leiomyosarcomas. Clinical implications. AB - We investigated the expression of P-glycoprotein (P-GP) and metallothionein (MT) in a series of 92 GIST and 14 gastrointestinal leiomyosarcomas (GILMS) with the purpose to expand our knowledge on the biological bases of GIST chemo-resistance and to ascertain their significance in patients' prognosis. P-GP expression was more frequent in GIST than in GI-LMS (83.7% vs. 21.4%, p<0.001), with no difference between low- and high-risk GIST (p=1.000) or low- and high-grade GI LMS (p=0.538). P-GP expression was unrelated to anatomic location (gastric vs. intestinal) in GIST (39/45 vs. 35/43, p=0.770) and in GI-LMS (0/2 vs. 2/6, p=1.000). MT expression was non-significantly higher in GI-LMS than in GIST (35.7% vs. 14.1%, p=0.060), with no difference between low- and high-risk GIST (p=1.000) or low- and high-grade GI-LMS (p=1.000). MT expression was unrelated to the anatomic location (gastric vs. intestinal) in GIST (7/45 vs. 6/43) and GI-LMS (0/2 vs. 1/6) (p=1.000 and p=0.1000, respectively). Overall tumor-specific survival (p< 0.001) and disease-free survival (p<0.001) were different in GIST as compared with GI-LMS, and the number of events was higher in GI-LMS. When the survival analysis took into consideration P-GP or MT expression, the overall survival in GIST was influenced by the expression of MT (p=0.021) but not by that of P-GP (p=0.638). However, in GI-LMS, P-GP expression influenced disease-free survival (p=0.050); in addition, it is important to recognize the limited value of these results because of the low number of cases involved in the study. Differential expression of P-GP and MT might explain the known variability in response to systemic chemotherapy in these tumors. Detection of P-GP and MT seems to add certain prognostic value in GIST (MT) or GI-LMS (P-GP). PMID- 17922050 TI - Optimization of PCR amplification for B- and T-cell clonality analysis on formalin-fixed and paraffin-embedded samples. AB - In many cases, particularly in retrospective studies, only formalin-fixed and paraffin-embedded (FFPE) tissue samples are available for molecular studies. DNA recovered from FFPE tissues generally consists of fragmented small target sequences with chemical alterations. Clonality analysis is not easy on FFPE samples, in fact, it requires even more experience than that of performed on fresh samples or is more complicated than most genomic PCR amplifications for somatic genes. In our study, we have performed a multi-parameter PCR evaluation investigating immunoglobulin heavy chain gene (IgH) and T-cell receptor gamma gene (TCRgamma) rearrangements on non-purified crude lysates of FFPE samples, in order to establish the significance of different variables on performance of PCR amplification. The results showed that a slight decrease in the concentration of primers in combination with a slight increase in MgCl2 and Taq polymerase concentrations, as well as the use diluted crude template and a standard amount of dNTPs can be the modifications of choice while adjusting IgH and TCRgamma clonality tests on poor quality DNA FFPE samples. Using our improved protocol, 74% (17/23) of the tested B-cell lymphomas and 68% (31/46) of the tested T-cell lymphomas demonstrated monoclonal PCR product, proving the applicability of our optimized method. Our experience may be of help during the optimization process in technically difficult cases as well as to determine which parameters and how should be changed to minimize false-negative and false-positive results. PMID- 17922051 TI - Relation of neuroendocrine cells to transforming growth factor-alpha and epidermal growth factor receptor expression in gastric adenocarcinomas: prognostic implications. AB - The presence of neuroendocrine (NE) cells in gastric adenocarcinoma (GCa) is well documented, however, their significance is controversial. There is no evidence in the literature concerning the possible effect of these cells on the expression of TGF-alpha and EGFR, which are believed to confer growth advantage to tumor cells. 101 partial or total gastrectomy specimens from patients operated for conventional gastric adenocarcinoma were included in the study. In each case immunohistochemistry was performed on sequential tissue sections for chromogranin A (ChrA), TGF-alpha and EGFR. Samples were graded based on the number of ChrA positive cells (0-3). TGF-alpha and EGFR expressions were evaluated according to both the intensity (0-2) and quantification of the positively stained areas (0 3). Follow-up data was available in 54 patients. Twenty-seven patients died of disease, while 27 patients were alive with a follow-up of at least 15 months. ChrA expression was detected in 54.4% of the tumor specimens. TGF-alpha was stained positively in 42.6% and EGFR in 49.5% of the cases. NE cells in GCa was related to TGF-alpha (p<0.0001) and EGFR expression (p<0.05), and TGF-alpha/EGFR coexpression (p<0.001). Among histopathologic variables, the presence of NE cells was significantly related to grade, stage and lymph node status. Although the presence of NE cells had no effect on survival, the expression of EGFR (p<0.0001) and TGF-alpha (p=0.002) were related to survival. The results of our study suggest that the presence of NE cells may have an effect on the expression of TGF alpha and EGFR in GCa, and the autocrine mechanism between TGF-alpha and EGFR plays an important role in the prognosis of gastric carcinoma. PMID- 17922052 TI - Complex organizational defects of fibroblast architecture in the mouse spleen with Nkx2.3 homeodomain deficiency. AB - The capacity of secondary lymphoid organs to provide suitable tissue environment for mounting immune responses is dependent on their compartmentalized stromal constituents, including distinct fibroblasts. In addition to various members of the tumor necrosis factor/lymphotoxin beta family as important morphogenic regulators of peripheral lymphoid tissue development, the formation of stromal elements of spleen is also influenced by the Nkx2.3 homeodomain transcription factor in a tissue-specific fashion. Here we extend our previous work on the role of Nkx2.3-mediated regulation in the development of spleen architecture by analyzing the structure of reticular fibroblastic meshwork of spleen in inbred Nkx2.3-deficient mice. Using immunohistochemistry and dual-label immunofluorescence we found both distributional abnormalities, manifested as poor reticular compartmentalization of T-zone and circumferential reticulum, and developmental blockade, resulting in the absence of a complementary fibroblast subpopulation of white pulp. The disregulated distribution of fibroblasts was accompanied with an increased binding of immunohistochemically detectable complement factor C4 by T-cell zone-associated reticular fibroblasts, distinct from follicular dendritic cells with inherently high-level expression of bound C4. These data indicate that the impact of Nkx2.3 gene deficiency on fibroblast ontogeny within the spleen extends beyond its distributional effects, and that the formation of various white pulp fibroblast subsets is differentially affected by the presence of Nkx2.3 activity, possibly also influencing their role in various immune functions linked with complement activation and deposition. PMID- 17922053 TI - Retrospective analysis of the prognostic role of tissue eosinophil and mast cells in Hodgkin's lymphoma. AB - The composition of reactive cell populations, which constitute the majority of tumor load in Hodgkin's lymphoma (HL), can influence the prognosis of the disease. Besides widely accepted and applied prognostic scores, the authors evaluate biological factors that may have a prognostic impact. Previous data indicate that the rate of eosinophils and mast cells in the reactive cell population, determined already at diagnosis, can be used for this purpose. Histological samples from 104 patients with HL with an average follow-up period of 110 (24-214) months were retrospectively analyzed. Mast cell positivity was associated with better overall survival, although this difference was only of borderline statistical significance (p=0.092). No significant difference was found in parameters like overall survival (OS, p=0.906) or event-free survival (EFS, p=0.307) of eosinophil-positive vs. -negative cases or in EFS (p=0.742) of mast cell-positive vs. -negative individuals (criterion for a positive specimen was more than 5% of appropriate cells in the reactive cell population). Looking at the effect of eosinophilia and mastocytosis together, there was no significant difference between the subgroups categorized according to the combined presence of the two cell types. It seems that tissue eosinophil and mast cell predominance have no prognostic value that could be used in clinical practice, although a tendency for correlation of mast cell positivity with overall survival could be seen. For a definitive statement, multicenter studies should be performed involving a higher number of patients suffering from HL. PMID- 17922055 TI - Anthracycline antibiotics induce acute renal tubular toxicity in children with cancer. AB - Experimental evidence suggests that anthracyclines, widely used in cancer chemotherapy, may impair kidney function. We assessed kidney function by serum creatinine, urinary N-acetyl-beta-D-glucosaminidase activity indices (NAGi) and microalbuminuria (MA) in 160 serum and urine samples obtained from 66 children with cancer. The effect of dexrazoxane was analyzed in 6 children on dexrazoxane supportive therapy in conjunction with daunorubicin (DNR) treatment, as compared with 6 children not receiving this agent. NAGi was significantly (p<0.05) elevated after treatment by DNR, doxorubicin, epirubicin (EPI) and idarubicin (IDA). MA proved to be a less sensitive indicator of kidney damage than NAGi. DNR resulted in a progressive deterioration of proximal tubular function as determined by linear regression analysis. The mean NAGi in the dexrazoxanetreated group was significantly (p<0.005) lower than in children not receiving dexrazoxane prior to DNR treatment. In conclusion, our study demonstrated that DNR, EPI and IDA induced an acute renal tubular damage similar to known tubulotoxic agents as cisplatin, carboplatin, cyclophosphamide and ifosfamide. The damage was clinically mild and only a minor proportion of patients can be expected to develop long-lasting tubulopathy with negative impact on the quality of life. PMID- 17922054 TI - Apolipoprotein A5 T-1131C variant confers risk for metabolic syndrome. AB - The -1131C is a naturally occurring variant of the apolipoprotein A5 (ApoA5) gene, which has been shown to associate with increased triglyceride levels. This variant has also been shown to confer risk for development of ischemic heart disease and stroke. The gene is in linkage disequilibrium with factors known to correlate with impaired glucose homeostasis. These observations prompted us to study the prevalence of the ApoA5 -1131C allele in patients with metabolic syndrome. A total of 201 metabolic syndrome patients and 210 controls were studied. In both groups the triglyceride levels of patients with -1131C allele were significantly increased compared to the subjects with -1131T allele (3.22+/ 0.43 mmol/l vs. 2.24+/-0.12 mmol/l, p<0.01 in the metabolic syndrome patients; 2.10+/-0.19 mmol/l vs. 1.22+/-0.05 mmol/l, p<0.01 in the controls). In metabolic syndrome patients the prevalence of the ApoA5 -1131C variant was increased compared to the healthy controls (11% vs. 6.20%). Multiplex regression analysis model adjusted for age, gender, serum total cholesterol levels, acute myocardial infarction and stroke events revealed that the examined ApoA5 variant confers risk for the development of metabolic syndrome: the odds ratio at 95% confidence interval was 3.622 (1.200-10.936), p=0.02. Our findings strongly suggest that this variant is a risk factor for the development of hypertriglyceridemia and metabolic syndrome. PMID- 17922057 TI - Primary bone marrow T-cell anaplastic large cell lymphoma with triple M gradient. AB - We present a case of a 60-year-old male patient with primary bone marrow anaplastic large cell lymphoma. He was admitted to the hospital with the symptoms of anemia and fever. There was no evidence of lymphadenopathy or splenomegaly. Immunoelectrophoresis showed the presence of a triple M gradient (double IgM and an IgG), with the IgG and one of the IgM paraproteins functioning as a cryoglobulin. The patient had no hepatitis C virus infection. Bone marrow biopsy showed massive CD30-positive, ALK-negative large lymphoid cell infiltration of T cell origin with anaplastic morphology. PCR analysis of lymphoid cells separated from the bone marrow demonstrated the presence of a B/T hybrid genotype disorder with no evidence of the t(2;5), nor t(1;2) translocations. The patient entered a period of remission following CHOP chemotherapy. The patient subsequently died of sepsis as a consequence of serious humoral immunodeficiency. PMID- 17922056 TI - Unique occurrence of Brachmann-de Lange syndrome in a fetus whose mother presented with a diffuse large B-cell lymphoma. AB - Brachmann-De Lange Syndrome (BDLS, MIM 122470) is a rare multiple congenital anomaly/mental retardation syndrome characterized by a variable phenotype including intrauterine fetal growth retardation, limb reduction and distinctive facial and skull features (low frontal hairline, synophrys, anteverted nostrils, long philtrum, downturned corners of the mouth, micro- and retrognathia, low-set ears and micro-/brachycephaly), as well as a significant psychological developmental delay. A proposed classification system for BDLS include a classic type with characteristic facial and skull changes, a mild type where similar changes may develop with time or may be partially expressed, and a third type including phenocopies, where phenotypic changes are casually related to chromosomal aneuploidies or teratogenic exposures. We report on a 22-week gestation fetus with BDLS, showing intrauterine fetal growth retardation, brachycephaly, micro-/retrognathia and monolateral single bone of the forearm, in a woman harboring diffuse large B-cell lymphoma. Meticulous family history was negative for malformations, syndromes, congenital anomalies or psychiatric disorders. There are very few reports of BDLS at early gestation, but to the best of our knowledge, this is the first case occurring simultaneously with a hematological neoplastic disease of the mother. PMID- 17922058 TI - [Financial aspects of targeted therapy of lung cancer as compared to conventional chemotherapy]. AB - Lung cancer is the leading cause of cancer death worldwide, and it is responsible for 20-25% of all cancer deaths. In Hungary, more than 4,000 lung cancer patients receive chemotherapy every year, and of them 3,000 suffer from non-small cell lung cancer. Despite the rapid development in antitumor treatment options, the response rates of chemotherapies in non-small cell lung cancer still remain between 15-35%. Recently, EGFR tyrosine kinase inhibitor therapy has been introduced with a response rate of 15% when used in unselected patients. However, this ratio may increase up to 70% when only patients with tumors containing EGFR mutation are treated. Therefore, results of translational research that could help pulmonologists and oncologists to apply tailored therapy in lung cancer patients are of great importance. The purpose of our work was to establish a model in order to study the relevance of immunohistochemical and molecular biological analyses of tumor specimens in treatment and patient selection, and to investigate their cost-sparing effects. We concluded that the most cost-effective type of patient selection could be achieved with EGFR mutation status analysis of the tumor tissue. Our results may help to determine the most cost-effective way of patient selection in case of non-small cell lung cancer patients requiring second line therapy; moreover, they might serve as a basis for further economic analyses. PMID- 17922059 TI - [Allocating resources for cancer control--resolving multicriteria decision-making using the analytic hierarchy process]. AB - When competing programs ought to be financed simultaneously for the same purpose, an allocation problem occurs due to scarce resources, and different perspectives and preferences. Facing the problem needs determining criteria which the decision might be based on. Those criteria form the objectives (the scope) of the different participants, and are relevant for the achievement of the goal, providing a comprehensive resource allocation that bridges and integrates the different perspectives. In case of cancer control primary prevention, secondary prevention, therapy and tertiary prevention, education, basic sciences, and clinical trials form the alternatives. An analytic hierarchy process (AHP) is used for supporting decision-making in the resource allocation problem. AHP is a method for setting priorities, but can only work out the implications of what was declared through the pairwise-ranking process, namely the relative preferences, weighing the criteria and rating the alternatives two by two. In the first analysis the relative weights to criteria were 0.099 for 'distributive justice'; 0.120 for constitutional and human rights; 0.251 for lay opinion; 0.393 for EBM; 0.137 for cost-effectiveness. Ranking the alternatives using 'judgements' resulted in relative preference of 0.238 for therapy, 0.204 for primary prevention, 0.201 for secondary prevention, 0.135 for clinical trials, 0.111 for tertiary prevention, 0.066 for basic sciences and 0.045 for education. In the second analysis the relative importance of "cost-effectiveness" was doubled, thus resulting in 0.234 for therapy, 0.216 for secondary prevention, 0.183 for primary prevention, 0.145 for clinical trials, 0.113 for tertiary prevention, 0.063 for basic sciences and 0.046 for education. Sensitivity analysis has shown that increasing the relative weight of cost-effectiveness up to approximately 0.4 changes the rank of alternatives, and above 0.4 this criterion gives secondary prevention preferences. According to the relative rates computed in both of the models all criteria vote for therapy, but these preferences change at the high level of weights, in case of EBM, 'rights', and cost-effectiveness. Cost effectiveness prefers secondary prevention to therapy; the criterion of constitutional and human rights and the criterion of evidence-based medicine vote for primary prevention. PMID- 17922060 TI - [Chemotherapy for advanced gastric cancer--our possibilities in 2007]. AB - Gastric cancer is the fourth most common malignancy in Hungary. Surgery remains the mainstay of any curative treatment, however, approximately two-thirds of patients diagnosed with gastric cancer have unresectable (locally advanced and/or metastatic) disease. In that stage, palliative chemotherapy is of crucial importance. Gastric cancer is heterogeneous regarding location, etiology, histology and natural course. This diversity is reflected in the results of randomized controlled trials as well. Patient selection, location of primary, stage and pretreatment of the tumor, as well as the reference treatment, patient stratification and endpoints have varied among trials. Based on the results of the clinical trials of the past decades, gastric cancer is chemosensitive, however, patient prognosis remains very poor, with a median survival of less than one year, and therapy-associated toxicity remains an issue. Based on the aforementioned, no standard regimens have yet been established worldwide. New compounds and targeted biological treatment make the development of more effective, less toxic, individualized treatment modalities possible. PMID- 17922061 TI - [Tumor movements detected by multi-slice CT-based image fusion in the radiotherapy of lung cancer patients]. AB - The aim of our study was to detect the possible uncertainties arising from tumor movements in the daily routine treatment planning, in extreme breathing conditions. Ten patients with lung cancer were enrolled into the study. According to tumor location, five patients had peripheral and five had central tumor. After the normal planning CT scan, two more scans were made with the same CT parameters in maximal exhalation and in maximal inhalation. For planning, the normal breathing scans were used with the fusion of the maximal inhalation and maximal exhalation scans. After the fusion in all breathing phases the gross tumor volumes were contoured (GTV1, GTV2, GTV3). Around the GTV1 (normal breathing phase GTV) 3 planning target volumes (PTV) were generated with the margin of 0.5 cm, 1.5 cm and 2.5 cm (PTV1, PTV2, PTV3). Individual plans were generated to all PTVs. All GTV volumes were registered. In all cases volume deviations were registered in different breathing phases (min: 1.5%, max: 35.6%). For GTV coverage comparison the coverage index (CI) was used. In case of extreme breathing conditions, using 0.5 cm margin was sufficient to reach good coverage for central tumors. For peripheral tumors 1.5 cm margin had to be used for the acceptable coverage (CI: 0.85-1.00). In our study, extreme breathing conditions were analyzed. According to our results, CT scans used in the daily routine do not exactly represent the tumor midposition and the true tumor volume. Due to breathing synchronous tumor movements, 0.5 cm margin must be used for planning in central location. In peripheral tumors wider margin should be used. PMID- 17922062 TI - [New diagnostic method in pulmonary carcinoid]. AB - In clinical oncology tumor markers could be helpful in diagnostics, and may play a complementary role beside the imaging systems in the follow up of patients with malignant disease. Chromogranin A can be considered as a marker in neuroendocrine malignancies such as pulmonary carcinoids. Based on recommendations adopted from the literature, the authors measured chromogranin A levels in sera of patients with carcinoid tumors of different stages. Although the patient number is low, our data suggest that in cases where carcinoid metastasis can be detected, chromogranin A levels are elevated. PMID- 17922063 TI - [Imaging of pediatric brain tumors using somatostatin analogue 111Ih-DTPA-D-Phe1 octreotide]. AB - Malignant solid tumors and leukemias are the second most common causes of death in childhood. The most frequent pediatric solid tumors are brain tumors. Brain tumors, especially medulloblastoma should be treated by surgery, irradiation and chemotherapy. However, chemotherapy has only moderate effect. Pediatric brain tumors, especially medulloblastomas, express somatostatin receptors. The aim of this study was the investigation of the expression of somatostatin receptors in pediatric brain tumors for diagnostic and therapeutic purpose. Fifty-six scintigraphic imagings (111In-DTPA-D-Phe1-octreotide) made in 45 children treated with brain tumor at the Unit of Oncology of the 2nd Department of Pediatrics, Semmelweis University. The diagnosis was medulloblastoma in 21 cases (46.7%). MRI scans have been performed parallel with the Octreoscan images. Octreoscan images were positive in 27 of 56 (48.2%) cases. The 27 positive Octreoscan images consisted of 16 medulloblastomas, 4 ependymomas, 4 astrocytomas and 3 glioblastomas. In 37 (66.1%) cases the results of Octreoscans were the same as those of the MRI scans. However, in 19 scans (33.9%) the outcome was different. Octreoscan imaging is not suitable for differential diagnosis in pediatric brain tumors, including medulloblastomas. Isotopes specifically binding to the somatostatin receptors (111In-DTPA-D-Phe1-octreotide) can be applied in medulloblastomas for diagnosis and follow-up treatment. In Octreoscan-positive tumors the Octreoscan images establish the opportunity to somatostatin analogue and/or specifically targeted radiation therapies. PMID- 17922064 TI - Acute and subchronic effects of gamma-hydroxybutyrate on isolation-induced aggression in male mice. AB - Gamma-hydroxybutyrate (GHB) is a new drug with abuse potential popularly known as "liquid ecstasy". It is an endogenous compound of the mammalian brain which satisfies many of the criteria for consideration as a neurotransmitter or neuromodulator. Preliminary studies have found that GHB (100-200 mg/kg) reduces aggressive behavior in mice. This study was designed to assess the effects of low and intermediate doses of GHB (5, 25, 50 and 100 mg/kg, ip) on isolation-induced aggression in male mice, using an ethopharmacological approach. Moreover, the possible development of tolerance after its subchronic administration for 15 consecutive days was also examined. Individually housed mice were exposed to anosmic "standard opponents" 30 min after drug administration, and the encounters were videotaped and evaluated using an ethologically based analysis. Acute treatment with GHB (25-100 mg/kg) significantly reduced offensive behaviors (threat and attack) without affecting immobility, whereas with the lowest dose used (5 mg/kg) a significant increase of attack behaviors was observed. This behavioral profile was maintained when GHB (25-100 mg/kg) was administered during 15 consecutive days, indicating an absence of tolerance to the initial antiaggressive action of the drug. However, the subchronic treatment with 5 mg/kg of GHB produced an opposite effect to that observed after single treatment, suggesting a possible desensitization of postsynaptic dopaminergic receptors. PMID- 17922065 TI - An effective dose of valdecoxib in experimental mouse models of pain. AB - The effects of selective cyclooxygenase-2 (COX-2) inhibitors in biological functions are frequently investigated in animal models. However, there is little data on their analgesic efficacy in experimental animals. This study aimed to determine whether oral gavage of 5 mg/kg valdecoxib in mice is active as an analgesic at this dose and whether it is associated with therapeutic blood levels. A nonselective COX inhibitor, ketorolac, was also investigated for comparison. A total of 106 C57 BL/6N mice were administered a single oral dose of 5 mg/kg of valdecoxib, ketorolac or placebo. The antinociceptive effects of both drugs were tested using hot-plate and formalin tests. For the hot-plate test, reaction time (latency) of the mouse before jumping was recorded. The total time that the mouse spent on licking/biting the injected paw (with dilute formalin) was recorded in the formalin test. Apart from the behavioral tests, plasma concentrations of the drugs at this dose were also determined. Mice were fed with 5 mg/kg of either valdecoxib or ketorolac. Blood samples were collected between 1 and 9 h postingestion. Valdecoxib and ketorolac concentration in the plasma was determined by high performance liquid chromatography with ultraviolet detection (HPLC-UV). Effective antinociception was observed for both drugs in the hot-plate test from 75 min to 2 h after oral dosing. Also, both drug treatments showed a significantly reduced nociceptive response in the second phase in the formalin test (20-30 min after injection). Both valdecoxib and ketorolac showed plasma concentrations comparable to the therapeutic concentrations in humans. A single oral dose of valdecoxib or ketorolac (5 mg/kg) is able to produce a therapeutic analgesic effect in mice. PMID- 17922066 TI - New approach for chronic renal failure model by direct kidney injection of doxorubicin in rats. AB - Several experimental chronic renal failure (CRF) models are available for testing new drugs. A CRF model induced by the intravenous injection of 2 mg/kg of doxorubicin (DXR) twice during a 20-day interval reportedly results in pathological characteristics similar to glomerular sclerosis seen clinically. However, it normally takes more than 16 weeks to create this CRF model. We used three methods of direct drug injection into the kidney of rats to determine the method that would induce CRF within 4 weeks; Method A: DXR was injected directly into both kidneys; Method B: DXR was injected directly into the left kidney immediately after right nephrectomy; Method C: DXR was injected directly into the left kidney 1 week before right nephrectomy, and DXR was injected again directly into the left kidney. As a result, urinary protein, blood urea nitrogen (BUN), creatinine and creatinine clearance were significantly changed >1 week after the injection of DXR by Method C. Quantification of tissue transforming growth factor beta1 (TGF-beta1), which is a prime fibrogenic cytokine in renal fibrosis, significantly increased in the kidney. A light microscopic image showed glomerular decrement, tubular dilation and atrophy and vacuolation of parenchyma. In conclusion, the results of this study demonstrate that the DXR model using Method C develops CRF within 4 weeks. PMID- 17922067 TI - Effects of ketamine on pulmonary TLR4 expression and NF-kappa-B activation during endotoxemia in rats. AB - The aim of the study was to investigate the effects of ketamine on pulmonary Toll like receptor 4 (TLR4) expression and nuclear factor kappa-B (NF-kappa-B) activity in rats with endotoxemia. Rats were treated with lipopolysaccharide (LPS, 5 mg/kg) to induce endotoxemia, followed immediately by ketamine (0.5, 5 and 10 mg/kg) or 0.9% NaCl (5 ml/kg). At 1, 3 and 5 h posttreatment, TLR4 mRNA expression and NF-kappa-B activity of the rat lungs were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and electrophoretic mobility shift assay (EMSA), respectively. LPS increased TLR4 expression and NF-kappa-B activity in the rat lungs. Ketamine at the dose levels of 5 and 10 mg/kg suppressed the LPS-induced elevation of TLR4 expression and NF-kappa-B activity. Ketamine alone had no effect on either one of them. The present study revealed that ketamine could inhibit pulmonary NF-kappa-B activity during endotoxemia in rats, and this antiinflammatory effect possibly acts through suppressing TLR4 expression. PMID- 17922068 TI - Changes in the plasma diazepam concentration and its anticonvulsant effect after the discharge of a diazepam suppository from the rectum in rats. AB - One of the clinical problems regarding the use of suppositories for patients is the discharge of the medication itself after the insertion of the suppository. The effects of the discharge of diazepam (DZP) suppositories from the rectum on the plasma DZP concentration and its anticonvulsant action were investigated in rats. The plasma DZP concentration reached a maximum at approx. 30 min after the rectal administration of a 5 mg/kg DZP suppository. The discharge of the DZP suppository from the rectum at 5 or 10 min after rectal administration significantly reduced the plasma DZP concentration and the anticonvulsant action of the DZP suppository against pentylenetetrazol-induced seizures. However, when the suppository was discharged from the rectum after 20 min, the plasma DZP concentration and the anticonvulsant action were only slightly but not significantly decreased. These results suggest that the discharge of the DZP suppository in the early phase, within 20 min, decreases the plasma concentration and the anticonvulsant action of the DZP suppository in rats. Therefore, when the suppository is discharged from the rectum immediately, sufficient observation and proper treatment are necessary. PMID- 17922069 TI - Estimation of intraocular pressure in rabbits using noncontact tonometer: a comparative evaluation with Schiotz tonometer. AB - Estimation of intraocular pressure in rabbits is often done by indentation tonometry using Schiotz tonometer. The use of Schiotz tonometer for estimation of intraocular pressure requires the use of topical anesthesia, poses risk of infection and injury to cornea and puts the animal under stress. Therefore, in this study noncontact tonometer was used to estimate intraocular pressure in rabbits and the technique is described for the first time. Twenty-five rabbits were subjected to intraocular pressure estimation using Schiotz as well as noncontact tonometer by two independent observers. All intraobserver and interobserver differences were within two standard deviation from the mean, indicating good repeatability and reproducibility of the two methods. A good linear correlation was observed between measurements done by Schiotz and noncontact tonometers with the correlation coefficient (r(2)) of 0.9122 and regression equation y = 1.632x + 4.8684. No significant difference was observed in percent change in intraocular pressure in timolol-treated, water-loaded rabbits using noncontact tonometer and Schiotz tonometer. Noncontact tonometer provides an easy, quick and safe method for intraocular pressure estimation, which is comparable to the conventional method of Schiotz tonometry. Therefore, despite the cost, noncontact tonometry seems to have a greater scope in experimental studies. PMID- 17922070 TI - Antistressor activity of Ocimum sanctum (Tulsi) against experimentally induced oxidative stress in rabbits. AB - Fresh leaves of Ocimum sanctum (O. sanctum) were evaluated for antistress activity against experimentally induced oxidative stress in albino rabbits. Animals of the test group received supplementation of 2 g fresh leaves of O. sanctum per rabbit for 30 days. Anemic hypoxia was induced chemically by injecting the rabbits with 15 mg sodium nitrite per 100 g body weight intraperitoneally. Results indicated that O. sanctum administration blunted the changes in cardiorespiratory (BP, HR, RR) parameters in response to stress. A significant (p < 0.01) decrease in blood sugar level was observed after 30 days of dietary supplementation of O. sanctum leaves. Significant increase (p < 0.05) in the levels of enzymatic (superoxide dismutase) and nonenzymatic (reduced glutathione) antioxidants was observed in the test group after the treatment with O. sanctum. Oxidative stress led to a lesser depletion of reduced glutathione (28.80%) and plasma superoxide dismutase (23.04%) in O. sanctum-treated rabbits. The results of this study suggest that the potential antistressor activity of O. sanctum is partly attributable to its antioxidant properties. PMID- 17922071 TI - RNA interference--about the reality to be exploited in cancer therapy. AB - The discovery of RNA interference (RNAi) in mammalian cells raises the expectations of gene therapy, especially in cancer. However, it is too early to say how great this promise may be because of many disputable problems including intracellular delivery of siRNA, the transient nature of RNAi and potential side effects after long-term treatment. The present microarray study demonstrates that the RNAi of one oncogene (encoding bcr-abl fusion protein in chronic myelogenous leukemia) triggers an overexpression of other "sleeping" oncogenes, antiapoptotic genes and factors, preserving the immortalization of bcr-abl-positive leukemia cells. PMID- 17922072 TI - Diurnal variation in the effect of angiotensin II on nociception. AB - The aim of this study was to investigate the effect of angiotensin II (Ang II) on nociception at particular time points of a 24-h cycle using different pain stimuli. A parallel investigation of phasic and tonic pain tests revealed different diurnal patterns of pain responses. Phasic pain test (mechanical paw pressure) in rats was characterized with shortest latencies during the dark phase, when the average of motor activity is greatest. Ang II (0.1 microg/animal) increased the latency of pain responses to mechanical and thermal stimulations mainly during the active dark phase. With regard to tonic pain, regardless of a weak circadian fluctuation of the number of pain responses (writhes) in mice, there was a tendency to attenuate the diurnal pattern of nociception. In contrast to the effect of Ang II on the phasic pain, it exerted an antinociceptive effect in the writhing test during the light phase. In summary, Ang II exerted an antinociceptive effect at the time points that have naturally high pain sensitivity. PMID- 17922073 TI - Gateways to clinical trials. AB - [111In]-DOTA-cG250, [131I]-Metuximab injection, [177Lu]-DOTA-cG250; Anatumomab mafenatox, AP-12009; BIBW-2992, Biricodar dicitrate; Cediranib, Cilengitide, Clevidipine, CNTO-528, CNTO-95, CP-870893; Disufenton sodium, DNK-333A; Ecallantide, Enzastaurin hydrochloride, Etravirine, Exatecan mesilate; Fingolimod hydrochloride; Human insulin, Hyaluronic acid; Indisulam, Inhaled insulin, Insulin glargine, Ipilimumab, Irofulven, Ispronicline, ITF-282; J591; KAI-9803; L Arginine hydrochloride, Laropiprant, LY-518674; Matuzumab, MB-7133, Methylnaltrexone bromide, MVA-5T4; Nemifitide ditriflutate; Obatoclax mesylate, Oral insulin; P-276-00, PF-562271, Picolinic acid; Quercetin; R-109339, R-547, Rivaroxaban, Ruboxistaurin mesilate hydrate; Seliciclib; Terameprocol, Tilarginine hydrochloride, Tolvaptan, Uracil; Vincristine. PMID- 17922075 TI - Complete mitochondrial DNA sequence of the scallop Placopecten magellanicus: evidence of transposition leading to an uncharacteristically large mitochondrial genome. AB - Complete sequence determination of the mitochondrial (mt) genome of the sea scallop Placopecten magellanicus reveals a molecule radically different from that of the standard metazoan. With a minimum length of 30,680 nucleotides (nt; with one copy of a 1.4 kilobase (kb) repeat) and a maximum of 40,725 nt, it is the longest reported metazoan mitochondrial DNA (mtDNA). More than 50% of the genome is noncoding (NC), consisting of dispersed, imperfectly repeated sequences that are associated with tRNAs or tRNA-like structures. Although the genes for atp8 and two tRNAs were not discovered, the genome still has the potential for encoding 46 genes (the additional genes are all tRNAs), 9 of which encode tRNAs for methionine. The coding portions appear to be evolving at a rate consistent with other members of the pectinid clade. When the NC regions containing "dispersed repeat families" are examined in detail, we reach the conclusion that transposition involving tRNAs or tRNA-like structures is occurring and is responsible for the large size and abundance of noncoding DNA in the molecule. The rarity of enlarged mt genomes in the face of a demonstration that they can exist suggests that a small, compact organization is an actively maintained feature of metazoan mtDNA. PMID- 17922076 TI - Convergent evolution of human and bovine haptoglobin: partial duplication of the genes. AB - Haptoglobin (Hp) is a hemoglobin-binding plasma protein consisting of two types of chains, called alpha and beta, which originate from a common polypeptide. In humans, but not in other mammals, Hp has been shown to occur in two allelic forms, Hp1 and Hp2, which differ in the length of the alpha-chain. The longer alpha-chain (in Hp2) seems to have arisen by an internal duplication of a gene segment coding for almost the entire alpha-chain of Hp1. In this article we show that Hp of cow (Bos taurus) contains an alpha-chain, the structure of which is similar to that of the human Hp2 alpha-chain. Furthermore, comparison of the structure of bovine Hp and human Hp2 suggests that the bovine gene arose by a duplication of the gene segment homologous to that duplicated in human Hp2. However, a phylogenetic analysis indicates that the two genes were formed independently. The evolutionary pressure that has led to the fixation of the Hps with a longer alpha-chain is not known. PMID- 17922074 TI - Signature of a primitive genetic code in ancient protein lineages. AB - The genetic code is the syntactic foundation underlying the structure and function of every protein in the history of the biological world. Its highly ordered degenerate complexity suggests an incremental evolution, the result of a combination of selective, mechanistic, and random processes. These evolutionary processes are still poorly understood and remain an open question in the study of early life on Earth. We perform a compositional analysis of ribosomal proteins and ATPase subunits in bacterial and archaeal lineages, using conserved positions that came and remained under purifying selection before and up to the most recent common ancestor. An observable shift in amino acid usage at these conserved positions likely provides an untapped window into the history of protein sequence space, allowing events of genetic code expansion to be identified. We identify Cys, Glu, Phe, Ile, Lys, Val, Trp, and Tyr as recent additions to the genetic code, with Asn, Gln, Gly, and Leu among the more ancient. Our observations are consistent with a scenario in which genetic code expansion primarily favored amino acids that promoted an increase in polypeptide size and functionality. We propose that this expansion would have been critical in the takeover of many RNA mediated processes, as well as the addition of novel biological functions inaccessible to an RNA-based physiology, such as crossing lipid membranes. Thus, expansion of the genetic code likely set the stage for the transition from RNA based to protein-based life. PMID- 17922077 TI - The novel alkylating prodrug J1: diagnosis directed activity profile ex vivo and combination analyses in vitro. AB - OBJECTIVE: The dipeptide J1 acts as a prodrug of melphalan with a significant increased potency in vitro resulting from activation by cellular aminopeptidases. The current study was performed to evaluate the ex vivo profile of J1 using 176 primary tumor cell cultures from patients. In addition, the activity of J1 in combination with eight standard drugs, representing different mechanistic classes, was studied in nine different human tumor cell lines of different histopathological origin. METHODS: Ex vivo evaluation of tumor type selectivity, was performed using the established fluorometric microculture cytotoxicity assay (FMCA). Combinations between J1 and eight standard chemotherapeutic drugs were analyzed using the median-effect method. RESULTS: The prodrug J1 expressed approximately 50- to 100-fold higher potency but similar activity profile as that of its metabolite, melphalan. The difference was greater in some diagnoses (e.g. breast cancer, NHL and AML), and exceptionally high in some breast cancer samples with aggressive phenotypes. Combination analysis of J1 and standard chemotherapeutics yielded several potentially additive and synergistic interactions, most striking for etoposide with significant synergism in all studied cell lines. CONCLUSIONS: In conclusion, the ex vivo profile suggests that further evaluation of J1 as the alkylating agent in for example aggressive breast cancer might be of particular interest, preferentially in combination with DNA topoisomerase II inhibitors like etoposide. PMID- 17922078 TI - Biotransformation and pharmacokinetics of the novel anticancer drug, SYUIQ-5, in the rat. AB - SYUIQ-5, a novel telomerase inhibitor, has demonstrated antitumor activity in nude mouse studies. The objective of the present study was to examine the metabolism and pharmacokinetics of SYUIQ-5 in rats. The plasma pharmacokinetics of SYUIQ-5 was nonlinear following i.p. administration at 15, 30 and 60 mg/kg. SYUIQ-5 metabolism in rat liver microsomes followed Michaelis-Menten kinetics, with Km and Vmax values of 12.3 microM and 2.01 nmol/min/mg protein, respectively. Ketoconazole significantly inhibited the metabolism of SYUIQ-5 in liver microsomes from rats pretreated with control vehicle or various inducers, whereas sulphaphenazole, ticlopidine, quinidine, and methylpyrazole had no inhibitory effects on SYUIQ-5 metabolism. Dexamethasone and beta-naphthoflavone (BNF), but not phenobarbital and ethanol, significantly induced SYUIQ-5 metabolism in rats. Alpha-naphthoflavone significantly inhibited SYUIQ-5 metabolism in liver microsomes from BNF-pretreated rats. Similar to other secondary amines, SYUIQ-5 underwent N-demethylation and O-oxygenation to at least two metabolites by rat liver microsomes. Pretreatment of rats with SYUIQ-5 at 0.1, 5 or 10 mg/kg for 5 days significantly induced the expression and activity of rat Cyp1A1/2, and induced Cyp3A1/2 expression at 10 mg/kg, but not Cyp2E1 and 2B1/2. These results indicate that that SYUIQ-5 exhibits dose-dependent pharmacokinetics in rats and it is mainly metabolized by Cyp3A1/2. PMID- 17922079 TI - The effects of heat treatment on selected properties of a conventional and a resin-modified glass ionomer cement. AB - This study investigated the effects of application of heat alone and heat & pressure on the compressive strength and modulus, the stress relaxation characteristics and the fluoride release of a conventional and a resin-modified glass ionomer cement. Cylindrical specimens were made from both materials and divided into 3 groups. One group was heat treated in an oven at 120 degrees C for 20 min, another group was subjected to heat & pressure at 120 degrees C for 20 min at 6-bar pressure. The third group acted as a control. The compressive strength and modulus, stress relaxation and fluoride release were tested over 56 days. The results of this investigation indicate that heat treatment had no significant effect on the conventional GIC used but significantly affected the resin modified GIC by increasing both the compressive strength and modulus and reducing the stress relaxation characteristics and the fluoride release. The use of GIC to produce inlay or onlay restorations that adhere to tooth tissue and release fluoride would be highly desirable. The results of this study indicate that it is possible to improve the strength of RMGIC with heat to a limited extent, but fluoride release may decrease. PMID- 17922080 TI - Evaluation of water sorption property and in vitro blood compatibility of poly(2 hydroxyethyl methacrylate) (PHEMA) based semi interpenetrating polymer networks (IPNs). AB - pH responsive smart biomaterials of gelatin and poly(2-hydroxyethyl methacrylate co-acrylic acid) were synthesized by redox polymerization and characterized by FTIR, Environmental Scanning Electron Microscopy (ESEM). The prepared environmental responsive biomaterials containing polyelectrolyte segments were assessed for their water sorption potential under varying experimental conditions. The diffusion mechanism of transport of water molecules arising due to solvent-polymer interaction was also analysed to predict the behaviour of continuously relaxing macromolecular chains. The in vitro blood compatibility of the prepared polymeric hydrophilic materials was evaluated by methods such as blood clot formation, platelet adhesion, percent haemolysis and protein adsorption study on the surface of the prepared biomaterials. PMID- 17922081 TI - Giant eccrine hidrocystoma presenting with unilateral ptosis and epiphora. AB - PURPOSE: To report the presentation and management of an atypical and advanced case of hidrocystoma of the eyelids with associated functional complications. METHODS: Interventional case report with clinical photographs at presentation, intra-operative photographs, and histology. RESULTS: A 55-year-old lady who presented with massive upper and lower eyelid swellings causing mechanical ptosis and epiphora, respectively, was managed by surgical excision of the lesions. Histology confirmed eccrine hidrocystoma. CONCLUSIONS: Eccrine hidrocystoma should be considered in the differential diagnosis of adnexal masses and such lesions may cause significant functional and cosmetic morbidity despite their histologically benign nature. PMID- 17922083 TI - Report of the JRAAC on the Situation of the Accreditation Training Programmes in Europe. PMID- 17922082 TI - Effect of endovascular hypothermia on acute ischemic edema: morphometric analysis of the ICTuS trial. AB - INTRODUCTION: Pilot studies of hypothermia for stroke suggest a potential benefit in humans. We sought to test whether hypothermia decreases post-ischemic edema using CT scans from a pilot trial of endovascular hypothermia for stroke. METHODS: Eighteen patients with acute ischemic stroke underwent therapeutic hypothermia (target = 33 degrees C) for 12 or 24 h followed by a 12-h controlled re-warm using an endovascular system. CT scans obtained at baseline, 36-48 h (right after cooling and re-warming) and 30 days were digitized, intracranial compartment volumes measured using a validated stereological technique, and the calculated change in CSF volume between the three time-points were used as an estimate of edema formation in each patient. Patients were grouped retrospectively for analysis based on whether they cooled effectively (i.e., to a temperature nadir of less than 34.5 degrees C within 8 h) or not. RESULTS: Eleven patients were cooled partially or not at all, and seven were effectively cooled. Baseline demographics and compartment volumes and densities were similar in both groups. At 36-48 h, the total CSF volume had significantly decreased in the not cooled group compared to the cooled group (P < 0.05), with no significant difference in mean volume of ischemia between them (73 +/- 73 ml vs. 54 +/- 59 ml, respectively), suggesting an ameliorative effect of hypothermia on acute edema formation. At 30 days, the difference in CSF volumes had resolved, and infarct volumes (73 +/- 71 ml vs. 84 +/- 102 ml, respectively) and functional outcomes were comparable. CONCLUSIONS: Endovascular hypothermia decreases acute post-ischemic cerebral edema. A larger trial is warranted to determine if it affects final infarct volume and outcome in stroke. PMID- 17922085 TI - [Clinical relevance of biomarkers in cancer related bone disease]. AB - Biomarkers as biochemical substances of collagen metabolism are produced during bone turnover and can be determined as parameters of bone metabolism not only in serum, but also in urine. These growth and decomposition products of the bone are already used to determine bone metabolism in osteoporosis and to prove efficacy of antiresorptive therapy. Metastases of the bone likewise show a higher rate of bone turnover. Nowadays detection of neoplastic bone lesions and progression of their spread are performed with x-rays, radionucleoide bone imaging and magnetic resonance imaging. In the future, biomarkers might improve early detection of bone lesions and follow-up of skeletal metastases. At present, the clinical use is documented insufficiently. In the foreseeable future the determination of the bone turnover markers and additional serum parameters of bone metabolism such as OPG, RANKL might be available for early diagnosis and follow-up in patients with bone metastatic diseases. PMID- 17922086 TI - [The role of hip protectors in the prevention of hip fractures in older people]. AB - Hip fractures are a serious health problem all over the world with grave consequences for the people and health systems involved. Osteoporosis and sideways falls and the force this involves in the hip area are the main cause for hip fractures. Hip protectors have been developed to prevent hip fractures, which are caused by falls. Biomechanical studies have shown their effectiveness, but compliance is low. Nevertheless, compliance can be raised by providing information on the dangers of hip fractures and the significance that hip protectors have to prevent this, as well as improving the design and mechanical properties of individual hip protectors. That is why clinical studies have produced controversial results. Future studies on hip protectors with a patient friendly design will clarify the situation. We present a review on the current situation and some future aspects. PMID- 17922087 TI - High-dose bisphosphonate therapy in an urgent case of spontaneous multiple vertebral fractures in a 55 year old woman. AB - An early postmenopausal Caucasian woman aged 55 sustained multiple vertebral fractures after a minor trauma. After exclusion of any kind of secondary osteoporosis, we administered due to clinical severity combined oral and cyclic intravenous bisphosphonate therapy (oral risedronate 35 mg/week, i.v. pamidronate 30 mg quarterly) with adequate calcium and vitamin D supplementation for 28 months. We performed a transiliac bone biopsy at baseline and at month 28. The paired samples were investigated by histomorphometry, by microCT-analysis for 3d structure and by qBEI representing bone mineral density distribution. Mineralisation of the bone matrix was not influenced by supplementation of calcium and vitamin D. Parameters of bone architecture and BMD improved; and a reduction of pain and increased mobility was observed. No further osteoporotic fractures occurred during the time of investigation. PMID- 17922088 TI - Physiological considerations about life cycles of universities. AB - In Austria, a new federal law concerning university reorganization (UG 2002) has led to the former university medical faculties in Vienna, Innsbruck and Graz becoming independent medical universities, despite warnings by competent persons that this could have negative consequences. One logical consequence for example, is a certain isolationist trend that will eventually lead to medicine withdrawing into a kind of "splendid isolation", physically separated, even, from the rest in brand new buildings. The splitting of each of the three Austrian universities, which included Medical Faculties, into a new Medical University and a 'Rest University', has led to the growth of two daughter cells each now larger in terms of administration than the former university. The historical development of universities in Europe has seen times of affluence and times of decline. The recent new laws will for a long period have a marked impact on medicine and on the Medical Universities. PMID- 17922089 TI - Bacterial contamination of solutions for parenteral administration for single- and multiple-dose vials after multiple use in the hospital. AB - Outbreaks traced to bacterial contamination of multiple-dose vials are reported in the literature. During a four-month period, multi-dose vials (MDVs), single dose vials (SDVs), and vials containing self-prepared admixtures were collected from various wards to analyse sterility of their contents. We examined 68 commercially available MDVs containing sodium chloride 0.9% or heparin with added preservative and 17 single dose vials (SDVs) containing aqua ad injectionem or sodium chloride 0.9% and 11 vials with admixtures (ADX) of heparin and sodium chloride 0.9%, both without preservative. Four of 96 (4.17%) vials were not sterile: two of them were contaminated with spore-forming bacteria, two with coagulase negative Staphylococci. Three of the samples were MDVs containing a preservative. The date of the first use was not marked on 28% of the vials. Twenty-eight samples were multiply used, although they were SDVs or ADXs without preservative or without an adequate amount of preservative, respectively. In 15 of 68 MDVs, the time limit after the first use was exceeded. On average, the volume of the samples was 80% of the original volume. A proportion of 4% of vials was not sterile. A training programme for health care workers in aseptic techniques and for validation of the preparation of solutions for parenteral use should be installed. PMID- 17922090 TI - [Indication of psychoanalytic treatment for personality disorders]. AB - AIM: The study investigates how a psychoanalytic institution reaches its aims by identifying different factors like diagnosis, sociodemographic characteristics of the patient, motivation of the psychoanalyst, and their impact on the indication process. MATERIAL AND METHODS: 38 patients diagnosed as having severe personality disorders and treated in a psychoanalytic outpatient clinic were compared with 49 patients who were referred for psychoanalytic treatment to a private practice. RESULTS: Although the groups were similar regarding the severity of their problems, more comorbidity and a more stable professional life were found in the group referred to private practice. Patients who tend to distort their social life in order to maintain a fragile psychic balance are more likely to get institutional treatment. The psychoanalytic team-conference and discussion of unconscious object-relationship pattern had great impact on the process of giving correct indications. CONCLUSION: The indication for psychoanalytic treatment does not correlate with personality disorder diagnosis or with the motivation of the therapist. Nevertheless, more differentiated, in clinical routine practicable instruments measuring reflective functioning, or the quality of object relationship have to be developed in order to give an adequate indication for psychoanalytic treatment. PMID- 17922091 TI - Focal progression in patients with gastrointestinal stromal tumors after initial response to imatinib mesylate: a three-center-based study of 38 patients. AB - BACKGROUND: This study aimed to investigate the outcome of patients with advanced gastrointestinal stromal tumors (GISTs) exhibiting focal disease progression during imatinib therapy, treated by surgical resection and imatinib continuation. METHODS: A consecutive series of 38 patients with metastatic GISTs who underwent treatment with imatinib at our centers during a defined period of time was evaluated. Patients were evaluated for demographics including tumor-related features, initial response, disease recurrence, and salvage treatment modalities, and were classified as having either focal or generalized progression upon presentation prior to salvage therapy. RESULTS: After a median follow-up of 31.8 months, 25 of the 38 (65.8%) patients had progressed. Nine (36%) patients were classified as having focal and 16 (64%) as having generalized progression. Salvage therapies were: surgical resection and imatinib dose escalation in patients exhibiting focal progression and imatinib dose escalation alone in the majority of patients exhibiting generalized progression. Focal progression was associated with prolonged progression-free survival (PFS) and overall survival (OS) after salvage therapy as compared with generalized progression (median PFS and OS, 11.3 months and not attained, versus 2.5 and 22.8 months, respectively). Six-month PFS was 89% and 39% in patients exhibiting focal and generalized progression, respectively. KIT mutation analysis of controlled and progressive lesions was performed in 4 patients with focal progression. Secondary KIT mutations affected progressive lesions, whereas nonprogressive lesions harbored the original mutations only. CONCLUSION: Patients with advanced GIST exhibiting focal disease progression during imatinib therapy may benefit from surgical resection and imatinib continuation. Imatinib resistance seems to be partial in these patients. PMID- 17922092 TI - Conditional survival in gastric cancer: a SEER database analysis. AB - BACKGROUND: Gastric cancer survival is typically reported in terms of survival from the time of diagnosis. Conditional survival is a more relevant measure of prognosis for patients who have already survived 1 or more years since diagnosis. METHODS: Using the Surveillance, Epidemiology, and End Results (SEER 17) database from the National Cancer Institute, we analyzed data from 20 018 gastric cancer patients diagnosed between 1988 and 1998. Using the life-table method, we computed 5-year relative conditional survival, grouped by summary stage, age, sex, and ethnicity, for patients who had already survived up to 5 years from diagnosis. RESULTS: Relative conditional survival improves over time for all groups of gastric cancer patients who survive a period of time after diagnosis. The largest gains in conditional survival were seen in patients with advanced stage disease. In general, females showed better survival than males. When grouped by ethnicity, Asians continued to have improved survival compared to other ethnic categories, even at 5 years out from diagnosis. CONCLUSION: For gastric cancer patients who survive a period of time after diagnosis, the largest increases in conditional survival are seen for patients with advanced stage disease and for those less than 65 years old. Conditional survival can provide more relevant prognostic information than survival from the time of diagnosis for gastric cancer patients who survive a period of time after diagnosis. PMID- 17922093 TI - Prognostic significance of gastrin expression in patients undergoing R0 gastrectomy for adenocarcinoma. AB - BACKGROUND: Gastrointestinal (GI) hormones regulate several GI functions, including the proliferation and repair of normal mucosa, and hormone receptors may therefore be implicated in the growth, invasion, and metastasis of cancers of the GI tract. The aim of this study was to determine the cellular distribution of gastrin in intestinal-type gastric cancers, and to determine its relationship to outcomes after R0 gastrectomy. METHODS: Eighty-six consecutive patients undergoing R0 gastrectomy for adenocarcinoma were studied. Normal gastric mucosa and tumor were stained for gastrin and their specific cellular distribution was determined. RESULTS: The duration of survival of patients whose tumors exhibited well-differentiated gastrin-positive tumor (GPT) cells (n = 12) was significantly poorer than that of patients whose tumors were GPT-negative (5-year survival, 30% vs 54%; P = 0.037). Patients with GPT-positive intestinal-type gastric cancer (5 of 47 patients) had the poorest survival of all (median, 14 months; 5-year survival, 0%; P = 0.006). In a multivariate analysis, only lymph node metastases (hazard ratio [HR], 2.13; 95% confidence interval [CI], 1.2 to 3.79; P = 0.01) and the presence of GPT cells (HR, 6.61; 95% CI, 1.74 to 25.09; P = 0.01) were independently and significantly associated with durations of survival in patients with intestinal-type gastric cancer. CONCLUSION: The presence of GPT cells in patients with gastric adenocarcinoma is a significant and independent prognostic indicator. PMID- 17922094 TI - Symptom evaluation of long-term postoperative outcomes after pylorus-preserving gastrectomy for early gastric cancer. AB - BACKGROUND: Since the early 1990s, pylorus-preserving gastrectomy (PPG) has been used in the treatment of patients with early gastric cancer in order to reduce postprandial symptoms. To date, there have been few reports of long-term symptom evaluation following this procedure. The aim of this study was to evaluate long term postoperative outcomes after PPG. METHODS: Three hundred and ninety-seven patients with early gastric cancer were enrolled in this study: 194 patients who underwent PPG and 203 who underwent distal gastrectomy with Billroth-I reconstruction (DGBI). We compared the symptoms for the two groups in a questionnaire on postoperative functional outcomes, endoscopy findings and the appearance of gallstones after surgery. RESULTS: The incidence of symptoms suggesting early dumping syndrome was significantly lower in the PPG group compared with the DGBI group (P < 0.05). The incidences of disturbed bowel habit and frequent flatus were significantly lower in the PPG than in the DGBI group. The average relative body weight (actual BW/ BW immediately before the surgery) was significantly better in the PPG than in the DGBI group (P < 0.001). CONCLUSION: The long-term results show that PPG has clear advantages over DGBI in terms of postoperative symptoms and functional outcomes. These results imply that PPG should be the recommended procedure for early gastric cancers located in the middle third of the stomach. PMID- 17922095 TI - Family history of cancer in Japanese gastric cancer patients. AB - The aim of this study was to evaluate the family history of cancer in Japanese gastric cancer patients and to investigate the clinicopathological features of gastric cancer patients with and without a family history of cancer. Four hundred and forty gastric cancer patients were enrolled in this study. The family history (first- and second-degree relatives) was investigated. The 440 patients were divided into three groups: (1) patients with a family history of gastric cancer; (2) patients with a family history of other cancers; and (3) patients without a family history of cancer. Two hundred and four patients (46.4%) reported a family history of cancer. Gastric cancer was the most frequent, with 98 patients having a total of 123 reports of gastric cancer in the family; colorectal cancer was the second most frequent and lung cancer was the third most frequent. The average ages of the group with a family history of gastric cancer and the group with a family history of other cancers were significantly lower than that of the patients without a family history of cancer. Other clinicopathological factors examined showed no significant difference between the groups. Japanese gastric cancer showed aggregation within second-degree relatives. The average age of the patients in the group with a family history of gastric cancer was the only significant factor that differed between gastric cancer patients with and without a family history of cancer. PMID- 17922096 TI - Esophagojejunostomy through minilaparotomy after laparoscopic total gastrectomy. AB - Although laparoscopic distal gastrectomy (LDG) has been accepted as a surgical option for the treatment of early gastric cancer, laparoscopic total gastrectomy (LTG) has been adopted less often, because a more difficult surgical technique is required for reconstruction. To reduce the technical difficulties, we made some modifications to the functional end-to-end anastomosis technique and performed esophagojejunal anastomosis through a minilaparotomy. First, for easier handling of the esophagus, the first application of the linear stapler to create the esophagojejunal anastomosis was performed before transection of the esophagus. Second, the jejunal limb was anastomosed to the left side of the esophagus, which, compared with the right side, made available more free space, sufficient to operate the stapling device. Third, to close the entry hole and complete the gastrectomy concurrently, a linear stapler was applied through the left lower trocar. With this technique, the closure of the access opening was performed easily and was monitored directly through the minilaparotomy. We successfully performed LTG with Roux-en-Y reconstruction using our modified procedure in seven patients without any anastomotic complications. We believe our procedure is a secure and reliable method for reconstruction after LTG and will facilitate adoption of LTG as a surgical option for patients with early upper gastric cancers. PMID- 17922097 TI - Laparoscopic esophagogastric circular stapled anastomosis: a modified technique to protect the esophagus. AB - Laparoscopic surgery is increasingly being applied to gastric cancer surgery, including proximal gastrectomy for the resection of cancer located in the upper gastric body. Despite the ease of use of stapling devices for end-to-end anastomosis, esophagogastric anastomosis is complicated by the narrow laparoscopic space, making the placement of an esophageal purse-string suture and anvil insertion into the fragile and contracted esophagus difficult. The aim of this study was to employ a novel esophagogastric anastomosis technique for laparoscopic surgery which may avoid esophageal breakdown. Eleven patients with early gastric cancer within the upper gastric body underwent laparoscopic proximal gastrectomy. The anvil of the stapler was introduced into the esophagus through a small gastrostomy, before transection of the esophagus. The esophageal to-anterior gastric wall anastomosis was performed using a double-stapling technique, without the need to apply a purse-string suture. The mean operation time was 237 +/- 15 min and estimated blood loss was 39 +/- 21 ml. The postoperative course was uneventful in all 11 patients, with no anastomotic leakage observed. Two patients needed endoscopic balloon dilation of an anastomotic stricture 24 to 28 days postoperatively. This modified procedure of laparoscopic esophagogastric anastomosis after proximal gastrectomy for the resection of cancer is a simple, rapid, and atraumatic technique which reduces the risk of anastomotic insufficiency. PMID- 17922098 TI - Experience of gastric cancer in a patient who had received a living-donor liver transplantation. AB - A 57-year-old woman had previously undergone a living-donor liver transplantation (LDLT) for end-stage liver disease related to hepatitis B virus. The liver graft had been donated by her husband. Her postoperative course had been uneventful. In the course of postoperative surveillance, she was incidentally found to have gastric cancer by an endoscopic examination 2 years after the liver transplantation. A gastric resection was the treatment choice, and the results were successful. The tumor, which was moderately differentiated adenocarcinoma, was limited to the mucosal layer, with no metastasis. In addition, a Helicobacter pylori infection was observed. This is the first reported case of a gastric cancer after LDLT. We report this case because of its importance regarding the need to carry out close surveillance in transplant recipients who are treated with immunosuppressive drugs, in order to make a timely identification of the occurrence of common malignancies. PMID- 17922099 TI - Advanced gastric cancer with a duplicated hepatic artery: preoperative diagnostic value of multidetector-row computed tomography for surgical resection. AB - A 63-year-old woman with appetite loss and general fatigue underwent gastrointestinal fiberscopy, which revealed type 2 advanced gastric cancer. Multidetector-row computed tomography revealed a massive gastric cancer invading the left hepatic lobe, pancreatic head, and common hepatic artery, as well as revealing a duplicated hepatic artery in which the right hepatic artery branched directly from the celiac axis, and ran behind the splenic vein. On the other hand, the common hepatic artery ran anterior to the splenic vein. We were able to perform pancreaticoduodenectomy with common hepatic artery resection and left lobectomy as curative surgery because her duplicated hepatic artery enabled us to ligate the common hepatic artery. Her postoperative clinical course was uneventful, and she is in good health 3 years after the surgery, without recurrence. We consider that multidetector-row computed tomography is very useful for the diagnosis of vascular anomaly, preoperative staging and decision making on the appropriate surgical strategy. PMID- 17922100 TI - Using product kernels to predict protein interactions. AB - There is a wide variety of experimental methods for the identification of protein interactions. This variety has in turn spurred the development of numerous different computational approaches for modeling and predicting protein interactions. These methods range from detailed structure-based methods capable of operating on only a single pair of proteins at a time to approximate statistical methods capable of making predictions on multiple proteomes simultaneously. In this chapter, we provide a brief discussion of the relative merits of different experimental and computational methods available for identifying protein interactions. Then we focus on the application of our particular (computational) method using Support Vector Machine product kernels. We describe our method in detail and discuss the application of the method for predicting protein-protein interactions, beta-strand interactions, and protein chemical interactions. PMID- 17922101 TI - Scanning electrochemical microscopy (SECM) as a tool in biosensor research. AB - Scanning electrochemical microscopy (SECM) is discussed as a versatile tool to provide localized (electro)chemical information in the context of biosensor research. Advantages of localized electrochemical measurements will be discussed and a brief introduction to SECM and its operation modes will be given. Experimental challenges of the different detection modes of SECM and its applicability for different fields in biosensor research are discussed. Among these are the evaluation of immobilization techniques by probing the local distribution of biological activity, the visualization of diffusion profiles of reactants, cofactors, mediators, and products, and the elucidation of (local) kinetic parameters. The combination of SECM with other scanning-probe techniques allows to maximize the information on a given biosensing system. The potential of SECM as a tool in micro-fabrication aiming for the fabrication of microstructured biosensors will be shortly discussed. PMID- 17922103 TI - The complexity of nectar: secretion and resorption dynamically regulate nectar features. AB - In this paper, we review the phenomenon of nectar resorption, focusing on its physiological and ecological meaning. Nectar resorption is a phenomenon that has long been known but was rarely reported until the 1990s. It has more recently been demonstrated in several species by various direct and indirect methodologies. It has generally been demonstrated in senescent flowers as a phenomenon separate in time from, and independent of, nectar secretion. The significance of this type of resorption is generally recognized as a resource recovery strategy, recycling at least some materials invested in nectar production. Nevertheless, nectar resorption can occur concomitantly with nectar secretion. Nectar production is therefore best considered as a unified process comprising nectar secretion and resorption. The modulation of these two opposite phases allows nectar concentration to be maintained in a range suitable for pollinators (nectar homeostasis). The mechanism of nectar resorption at the cell level has received little attention, and its molecular basis can only be hypothesized on the basis of recent studies concerning sugar sensing. PMID- 17922102 TI - beta-cell Regeneration: neogenesis, replication or both? AB - Both type I and type II diabetes are characterized by beta-cell loss and dysfunction. Therefore, a major goal of diabetes therapy is to promote the formation of new beta-cells, either in vitro for transplantation or in vivo, i.e., beta-cell regeneration. The question of whether beta-cell regeneration occurs by replication of preexisting beta-cells or by neogenesis from a precursor within the pancreas is a major focus of interest. Lineage-tracing studies have found evidence only for beta-cell replication, while earlier studies based upon the appearance of insulin-positive cells in areas outside of islets formed the basis for the belief that neogenesis from precursors can occur in adult animals. Recently, we found that nonendocrine pancreatic epithelial cells could be induced to undergo endocrine differentiation under the influence of inductive factors from the human fetal pancreas. One possibility is that, similar to models of hepatocyte regeneration, beta-cells can arise either by neogenesis or replication, depending on the particular stimulus. Clearly, understanding the nature and control of beta-cell regeneration is critical for success in efforts to treat diabetes by beta-cell replacement. PMID- 17922104 TI - Notch signalling suppresses apoptosis in adult human and mouse pancreatic islet cells. AB - AIMS/HYPOTHESIS: The pathogenesis of diabetes and the success of islet transplantation depend on the control of pancreatic beta cell fate. The Notch signalling pathway is essential for normal prenatal pancreatic development, but the presence and function of this gene network in adult islets has received much less attention. METHODS: The presence of Notch signalling components was assessed in vitro using RT-PCR, western blotting and immunofluorescence. The functional consequences of altering Notch signalling on insulin secretion and programmed cell death were examined. RESULTS: Adult mouse islets, human islets and mouse insulinoma MIN6 cells possess key components of the Notch pathway. RT-PCR, western blotting and immunofluorescence indicated that the Notch target gene, neurogenin3 (Ngn3, also known as Neurog3), is also present in adult islet cells. Inhibiting Notch signalling with N-[N-(3,5-difluorophenacetyl-L-alanyl)]-S phenylglycine t-butyl ester (DAPT) increased Ngn3 mRNA expression and protein levels in adult islets. The activated notch homologue 1 (NOTCH1) protein level was decreased upon serum withdrawal, as well as after treatment with a phosphatidylinositol 3-kinase inhibitor, or hydroxy-2 naphthalenylmethylphosphonic acid, an insulin receptor inhibitor. While islets cultured in DAPT did not exhibit defects in insulin secretion, indicating that differentiation is unaltered, inhibiting gamma-secretase-dependent Notch activation led to a dose-dependent increase in caspase-3-dependent apoptosis in both MIN6 cells and human islets. Conversely, gamma-secretase overactivity resulted in an accumulation of cleaved NOTCH1 and protection from apoptosis. CONCLUSIONS/INTERPRETATION: Together these results show that the Notch/Ngn3 signalling network is intact and functional in adult islets. This pathway represents an attractive target for modulating beta cell fate in diabetes, islet transplantation and efforts to derive beta cell surrogates in vitro. PMID- 17922105 TI - Increased nicotinamide nucleotide transhydrogenase levels predispose to insulin hypersecretion in a mouse strain susceptible to diabetes. AB - AIMS/HYPOTHESIS: Insulin hypersecretion may be an independent predictor of progression to type 2 diabetes. Identifying genes affecting insulin hypersecretion are important in understanding disease progression. We have previously shown that diabetes-susceptible DBA/2 mice congenitally display high insulin secretion. We studied this model to map and identify the gene(s) responsible for this trait. METHODS: Intravenous glucose tolerance tests followed by a genome-wide scan were performed on 171 (C57BL/6 x DBA/2) x C57BL/6 backcross mice. RESULTS: A quantitative trait locus, designated hyperinsulin production-1 (Hip1), was mapped with a logarithm of odds score of 7.7 to a region on chromosome 13. Production of congenic mice confirmed that Hip1 influenced the insulin hypersecretion trait. By studying appropriate recombinant inbred mouse strains, the Hip1 locus was further localised to a 2 Mb interval, which contained only nine genes. Expression analysis showed that the only gene differentially expressed in islets isolated from the parental strains was Nnt, which encodes the mitochondrial proton pump, nicotinamide nucleotide transhydrogenase (NNT). We also found in five mouse strains a positive correlation (r2 = 0.90, p < 0.01) between NNT activity and first-phase insulin secretion, emphasising the importance of this enzyme in beta cell function. Furthermore, of these five strains, only those with high NNT activity are known to exhibit severe diabetes after becoming obese. CONCLUSIONS/INTERPRETATION: Insulin hypersecretion is associated with increased Nnt expression. We suggest that NNT must play an important role in beta cell function and that its effect on the high insulin secretory capacity of the DBA/2 mouse may predispose beta cells of these mice to failure. PMID- 17922106 TI - Continuous and intermittent cardiac output measurement in hyperdynamic conditions: pulmonary artery catheter vs. lithium dilution technique. AB - OBJECTIVE: This study aimed to assess the level of agreement of both intermittent cardiac output monitoring by the lithium dilution technique (CO(Li)) and continuous cardiac output monitoring (PulseCO(Li)) using the arterial pressure waveform with intermittent thermodilution using a pulmonary artery catheter (CO(PAC)). DESIGN: Prospective, single-center evaluation. SETTING: University Hospital Intensive Care Unit. PATIENTS: Patients (n=23) receiving liver transplantation. INTERVENTION: Pulmonary artery catheters were placed in all patients and CO(PAC) was determined using thermodilution. CO(Li) and PulseCO(Li) measurements were made using the LiDCO system. MEASUREMENTS AND MAIN RESULTS: Data were collected after intensive care unit admission and every 8h until the 48th hour. A total of 151 CO(PAC), CO(Li) and PulseCO(Li) measurements were analysed. Bias and 95% limit of agreement were 0.11lmin(-1) and -1.84 to + 2.05 lmin(-1) for CO(PAC) vs. CO(Li) (r=0.88) resulting in an overall percentage error of 15.6%. Bias and 95% limit of agreement for CO(PAC) vs. PulseCO(Li) were 0.29 lmin(-1) and -1.87 to + 2.46 lmin(-1) (r=0.85) with a percentage error of 16.8%. Subgroup analysis revealed a percentage error of 15.7% for CO(PAC) vs. CO(Li) and 15.1% for CO(PAC) vs. PulseCO(Li) for data pairs less than 8 lmin(-1), and percentage errors of 15.5% and 18.5% respectively for data pairs higher than 8 lmin(-1). CONCLUSION: In patients with hyperdynamic circulation, intermittent and continuous CO values determined using the LiDCO system showed good agreement with those obtained by intermittent pulmonary artery thermodilution. PMID- 17922107 TI - Urinary bladder leiomyosarcoma in adults. AB - INTRODUCTION: Non-urothelial neoplasms of the bladder account for fewer than 5% of all bladder tumors. Sarcoma constitutes the most usual mesenchymal malignancy of the bladder, with leiomyosarcomas being the most common type of sarcoma in adults. PATIENTS AND METHODS: The records of seven patients presenting to two different institutions with bladder leiomyosarcomas between 2003 and 2007 and between 2000 and 2007, respectively, were examined. Cystoscopy, with transurethral resection of the bladder tumor was initially performed in all patients, with leiomyosarcoma being initially diagnosed on the basis of examination of the transurethral specimen. RESULTS: There were N = 5 men and N = 2 women with a median age of 64.8 years at presentation. All seven patients underwent a definitive surgical procedure. Complete resection with negative surgical margins was achieved in all seven patients (100%). MSKCC stage included 86% of patients with stage 3 (N = 6) and 14% with stage 2 (N = 1). A low-grade tumor was evident in one patient only, with the remaining 86% exhibiting a high grade tumor. DISCUSSION: Leiomyosarcomas of the bladder have always been considered as a highly aggressive entity and little is known about their origin, clinicopathologic presentation, and the survival factors associated with them. Contemporary studies suggest that these tumors may have a better prognosis than once believed. Leiomyosarcomas require aggressive surgical extirpation and, when surgical resection is possible, radical cystectomy with wide margins is the rule and should be performed. Strict adherence to standard surgical technique has resulted in low rates of positive surgical margins and low rates of local tumor recurrence. PMID- 17922108 TI - Clinical outcomes of elderly patients undergoing chronic peritoneal dialysis: experiences from one center and a review of the literature. AB - OBJECTIVES: The purpose of this study was to evaluate the outcome and to identify predictors of mortality in elderly patients on chronic peritoneal dialysis (CPD). METHODS: We retrospectively reviewed the charts of patients who started on CPD at the Division of Nephrology, University Health Network (UHN), Toronto, from 1 January 1994 to 31 December 2001. Patients were divided into three different age groups (/=75 years). Baseline variables included demographics, information on primary kidney disease, comorbidities when dialysis was first started, and initial biochemical data such as serum albumin, serum calcium (corrected for protein), phosphate, hemoglobin (Hb), total cholesterol, and triglyceride. The effects of these variables on survival were studied using a univariate procedure and then analyzed using multivariate Cox proportional hazards models in order to evaluate their independent relation to mortality. RESULTS: This study included 358 patients, among whom 213 (59.5%) were /=75 years old. Mean actuarial (death-censored) technique survival for the overall study population was 72.4 months (95% confidence interval [CI]: 66.3-78.5); in the /=75 year-old groups mean survivals were 74.4, 62.0, and 64.5 months, respectively. The death-censored technique survival for the elderly patients was not statistically significantly different from that in young patients (P = 0.778). In the overall study population, the mean patient survival was 70.4 months (95% CI 64.2-76.6), while the mean survivals for the /=75 year-old groups were 82.3, 54.0, and 50.0 months, respectively. The overall survival rates at 12 months were 98%, 84%, and 85% for the /=75 year-old groups, respectively. Not surprisingly, the survival of elderly patients on CPD is shorter than that of younger patients (P = 0.000). There were no significant differences between the two elderly groups (P = 0.439). Mortality was predicted by lower initial serum total cholesterol and albumin as well as higher serum calcium levels. CONCLUSION: Our study shows that elderly patients starting CPD had a death-censored technique survival comparable to that of younger patients. As expected, the survival of elderly patients on CPD was shorter than the survival of younger patients. Lower initial serum total cholesterol and albumin as well as higher initial serum calcium were associated with mortality in the elderly population. Our findings indicate that chronic peritoneal dialysis is a successful dialysis option for elderly patients with end stage renal disease. Measures to improve their nutritional state and achieve normalization of serum calcium might improve their survival. PMID- 17922110 TI - Quetiapine in primary insomnia: a pilot study. PMID- 17922109 TI - Porphyromonas gingivalis HmuY and HmuR: further characterization of a novel mechanism of heme utilization. AB - Porphyromonas gingivalis HmuY is a putative heme-binding lipoprotein associated with the outer membrane. It is part of an operon together with a gene encoding an outer-membrane hemin utilization receptor (HmuR) and four uncharacterized genes. A similar operon organization was found in Bacteroides fragilis and B. thetaiotaomicron, with the former containing an additional HmuY homologue encoded upstream of the hmuR-like gene. In P. gingivalis cultured under heme-limited conditions, a approximately 1-kb hmuY transcript was produced at high levels along with some approximately 3.5 and approximately 9-kb transcripts. Compared with the parental strain, mutants deficient in hmuY or hmuR or hmuY-hmuR gene function grew more slowly and bound lower amounts of hemin and hemoglobin. Significantly, they grew more slowly or were unable to grow when human serum was used as the sole iron/heme source. Analysis of the hmu promoter showed that it is regulated by iron. The HmuY protein normally occurs as a homodimer, but in the presence of hemin it may form tetramers. These results show that HmuY may be the first reported member of a new class of proteins in Porphyromonas and Bacteroides species involved in heme utilization, a function being exerted in conjunction with HmuR, an outer-membrane heme transporter. PMID- 17922112 TI - Age- and sex-dependent amphetamine self-administration in rats. AB - RATIONALE: Recreational drug use peaks in the developmental stage of adolescence, and exposure to drugs during adolescence may predict drug dependence in adulthood. Nevertheless, adolescent drug vulnerability is not widely studied in animal models of drug intake, and very few studies have investigated sex differences in drug-related behavior during adolescence. OBJECTIVES: We compared patterns of intravenous (i.v.) amphetamine self-administration among adolescent vs adult, male vs female Sprague-Dawley rats on a fixed ratio (FR) followed by a progressive ratio (PR) schedule of reinforcement. MATERIALS AND METHODS: After surgical implantation of i.v. catheters, adolescent [postnatal day (P) 35-52] and adult (P90-106) male and female rats were allowed to acquire lever-pressing behavior reinforced by either 0.025 or 0.05 mg/kg/0.1-ml amphetamine infusions over 14 daily 2-h sessions on an FR1 schedule (n = 9-12 per age-, sex-, and dose group). Subsequently, responding maintained by 0.0125 or 0.05 mg/kg per infusion amphetamine in 4-h sessions on a PR schedule was tested. RESULTS: Adolescent rats acquired amphetamine self-administration faster than adults, reached a higher number of infusions, and took more amphetamine than their adult counterparts during the acquisition phase, although age differences varied by dose. In PR testing, young adult males earned fewer infusions than older adult males, whereas young adult females earned more infusions than their older adult counterparts, and more than age-matched males. CONCLUSION: These results suggest that i.v. amphetamine self-administration in rats is a useful model to investigate the potential neurochemical and endocrine bases for age and sex differences in vulnerability to behavioral reinforcement by amphetamine. PMID- 17922113 TI - Metabolic profiling using Fourier-transform ion-cyclotron-resonance mass spectrometry. AB - With the aid of the extreme resolving power of Fourier-transform ion-cyclotron resonance mass spectrometry (FT-ICR/MS), we have developed a metabolomics platform for high-throughput metabolic profiling and metabolite candidate identification integrating a data-processing system, the Dr.DMASS program ( http://kanaya.naist.jp/DrDMASS/ ), and a metabolite-species database, KNApSAcK ( http://kanaya.aist-nara.ac.jp/KNApSAcK/ ). We discuss the potential of this FT ICR/MS-based metabolic profiling scheme as a general metabolomics tool by clarification of plant metabolic disorders and specific metabolite accumulation patterns caused by herbicidal enzyme inhibitors. PMID- 17922111 TI - Pro-cognitive effects of 5-HT6 receptor antagonists in the social recognition procedure in rats: implication of the frontal cortex. AB - RATIONALE: 5-HT6 receptor antagonists improve cognitive processes in rodents. However, their site(s) of action remains unexplored and their influence upon social memory has been little investigated. OBJECTIVES: We examined the influence of 5-HT6 receptor ligands upon social memory in rats by use of systemic or local administration into the frontal cortex (FCX), striatum, or nucleus basalis magnocellularis (NBM). MATERIALS AND METHODS: The social recognition test is based upon the ability of an adult rat to recognize a younger conspecific during the second of two 5-min sessions. In a procedure without an inter-session interval, the actions of drugs alone and the ability to reverse "amnesia" induced by the muscarinic antagonist, scopolamine (1.25 mg/kg, s.c.), were examined. The potential promnesic effect of drugs was also investigated in another procedure where a spontaneous deficit of recognition was induced by a 120-min inter-session interval. RESULTS: The 5-HT6 receptor agonist, WAY-181187 (10.0 mg/kg, i.p.), significantly impaired social recognition. This effect was abolished by the 5-HT6 receptor antagonists, SB-271046 (20.0 mg/kg, i.p.) and SB-258585 (10.0 mg/kg, i.p.). These agents also abolished scopolamine-induced amnesia (10.0 and 2.5 mg/kg, i.p., respectively) and reversed the delay-induced deficit (10.0-20.0 and 2.5-10.0 mg/kg, i.p., respectively). WAY-181187 into the FCX significantly impaired social recognition (0.16-0.63 microg/side). Conversely, SB-271046 into the FCX (2.5-5.0 microg/side), but neither into the striatum nor the NBM, significantly reversed spontaneous deficit. CONCLUSION: These results indicate that 5-HT6 receptors modulate social recognition by actions in the FCX and underpin their pertinence as targets for the treatment of psychiatric disorders in which cognitive function is compromised. PMID- 17922114 TI - Discrimination of biofilm samples using pattern recognition techniques. AB - Biofilms are complex aggregates formed by microorganisms such as bacteria, fungi and algae, which grow at the interfaces between water and natural or artificial materials. They are actively involved in processes of sorption and desorption of metal ions in water and reflect the environmental conditions in the recent past. Therefore, biofilms can be used as bioindicators of water quality. The goal of this study was to determine whether the biofilms, developed in different aquatic systems, could be successfully discriminated using data on their elemental compositions. Biofilms were grown on natural or polycarbonate materials in flowing water, standing water and seawater bodies. Using an unsupervised technique such as principal component analysis (PCA) and several supervised methods like classification and regression trees (CART), discriminant partial least squares regression (DPLS) and uninformative variable elimination-DPLS (UVE DPLS), we could confirm the uniqueness of sea biofilms and make a distinction between flowing water and standing water biofilms. The CART, DPLS and UVE-DPLS discriminant models were validated with an independent test set selected either by the Kennard and Stone method or the duplex algorithm. The best model was obtained from CART with 100% correct classification rate for the test set designed by the Kennard and Stone algorithm. With CART, one variable describing the Mg content in the biofilm water phase was found to be important for the discrimination of flowing water and standing water biofilms. PMID- 17922115 TI - Development of a colloidal gold-based lateral-flow immunoassay for the rapid simultaneous detection of zearalenone and deoxynivalenol. AB - A multianalyte lateral-flow technique using colloidal gold-labeled monoclonal antibodies was developed for the rapid simultaneous detection of deoxynivalenol (DON) and zearalenone (ZEA). The results of this qualitative one-step test were interpreted visually. A very simple and fast sample preparation was used, and the assay procedure could be accomplished within 10 min. When applied to spiked wheat samples, the technique gave accurate and reproducible results. Cut-off levels of 1500 and 100 microg kg(-1) for DON and ZEA, respectively, were observed. The described multianalyte format can be used as a reliable, rapid and cost-effective on-site screening technique for the simultaneous determination of mycotoxins in grain samples. PMID- 17922116 TI - Specific immunoassays for endocrine disruptor monitoring using recombinant antigens cloned by degenerated primer PCR. AB - Vitellogenin (VTG) and choriogenin (CHO) are valuable biomarkers of endocrine disrupting compound (EDC) exposure in fish. Existing immunoassays are limited to a few species, which restricts their use for the analysis of local wildlife sentinels. Using C. facetum as a relevant South American model fish, this work presents a new strategy for the preparation of antibodies to VTG and CHO, with zero cross-reactivity with fish serum components. Recombinant fragments of Cichlasoma facetum VTG (280-mer) and CHO (223-mer) were prepared by degenerate primer RT-PCR and expression in E. coli. Polyclonal and monoclonal antibodies prepared with these antigens were used to develop rapid dotblot assays for VTG and CHO. Both the polyclonal and monoclonal antibodies prepared with the recombinant antigens reacted against the native proteins adsorbed on to nitrocellulose allowing the set up of sensitive dotblot assays. The VTG assay was further validated with spiked samples and purified native VTG. Exposure experiments with several estrogenic compounds revealed the potential of C. facetum as a sensitive biomonitor that produced measurable responses at concentrations of 100 ng L(-1) of 17-beta-estradiol, 100 ng L(-1) of ethynylestradiol, and 6.6 microg L(-1) of nonylphenol. The approach described here may be applied to other native species to produce highly specific and sensitive rapid tests. It may be particularly advantageous for species that cannot be kept in captivity or when homogeneous purification of the immunizing proteins is particularly challenging. In conclusion, we present a novel approach to develop a strategy for the generation of immunoassay reagents for vitellogenin (VTG) and choriogenin (CHO), which will facilitate regional studies on the impact of endocrine-disrupting chemicals on local wildlife. PMID- 17922118 TI - Flexibility and individual differences in visuo-proprioceptive integration: evidence from the analysis of a morphokinetic control task. AB - Conflicting theories of visuo-proprioceptive integration in movement control suggest that each modality can be weighted according to either its statistical reliability or the computations in which the integrated estimates will be used. However, the psychophysical experiments on which most studies are based use sensory conflicts and are therefore likely to reflect particular rather than normal behavior. In this paper, we: (1) propose a method avoiding the use of sensory conflicts (delayed recall task), (2) restrict our interest to spontaneous rather than adapted behavior, and (3) focus on a complex task requiring fine online control in order for hand movements to fit a precise path during execution. Subjects were provided with either visual, proprioceptive or both cues while their right hand was passively moved to fit a precise three segment pathway. As soon as this encoding phase ended, they were instructed to reproduce actively the trajectory, either with vision, proprioception or both cues. Results provide evidence that vision and proprioception may be used very differently, (1) not only according to the relative resolution of the sensory systems in the actual context, (2) not only according to the processes involved in the task, but also (3) according to subjects. They also suggest that visual and proprioceptive cues are not fused to provide a weighted average position, but that the non dominant cue could simply be ignored when subjects are provided with multiple sensory cues. We conclude that each of these observations illustrates the same fundamental property of flexibility of integrative mechanisms. PMID- 17922119 TI - Asynchrony from synchrony: long-range gamma-band neural synchrony accompanies perception of audiovisual speech asynchrony. AB - Real-world speech perception relies on both auditory and visual information that fall within the tolerated range of temporal coherence. Subjects were presented with audiovisual recordings of speech that were offset by either 30 or 300 ms, leading to perceptually coherent or incoherent audiovisual speech, respectively. We provide electroencephalographic evidence of a phase-synchronous gamma oscillatory network that is transiently activated by the perception of audiovisual speech asynchrony, showing both topological and time-course correspondence to networks reported in previous neuroimaging research. This finding addresses a major theoretical hurdle regarding the mechanism by which distributed networks serving a common function achieve transient functional integration. Moreover, this evidence illustrates an important dissociation between phase-synchronization and stimulus coherence, highlighting the functional nature of network-based synchronization. PMID- 17922117 TI - Searching for two feature singletons in the visual scene: the localized attentional interference effect. AB - The localized attentional interference (LAI) effect was investigated in a visual search task requiring participants to simultaneously monitor two spatially separated features from the same or different dimensions. In Experiment 1, the search type was blocked and targets were defined by fixed feature values in two dimensions (e.g., a yellow item and a circular item). In contrast, in Experiment 2, participants had to look for a color and a form singleton, with the exact feature values varying randomly across trials. In both experiments, reaction times (RTs) were generally slower when two features were CLOSE to, rather than DISTANT from, each other. Moreover, RTs to CLOSE stimuli increased as the search set size increased, while RTs to DISTANT stimuli were unaffected by set size. Experiment 3 also used a singleton search task, but with the two singletons defined either in different dimensions or in the same dimension. A larger interference effect for CLOSE, as compared to DISTANT, stimuli was found for cross-dimension than for intra-dimension targets. These findings suggest that neighboring items, irrespective of whether these items are from the same or different dimensions, interfere with each other in attentional selection, and that searching for two cross-dimension targets may engage a process of dimension switching to effectively solve the ambiguity of each item, especially when these items are close to each other. PMID- 17922120 TI - Acute bilateral basal ganglia lesions in diabetic uraemia: diffusion-weighted MRI. AB - METHODS: We studied four patients with diabetes mellitus and chronic renal failure who developed sudden choreic movement disorders. The clinical manifestations, laboratory findings, MR imaging findings, and clinical outcome in each patient were evaluated. RESULTS: All four patients had long-term diabetes mellitus and severe azotaemia. Brain MR findings consisted of bilateral symmetric basal ganglia lesions, with decreased signal intensity on T1-weighted images and increased signal intensity on T2-weighted images. All three patients who underwent diffusion-weighted MR imaging (DWI) showed signal intensities similar to those of the surroundings in regions corresponding to increased signal intensity on T2-weighted images, with slightly increased apparent diffusion coefficient (ADC) values. Two of the patients showed small focal areas of restricted diffusion within the basal ganglia lesions. After haemodialysis, follow-up MR imaging in all patients demonstrated that the basal ganglia lesions had regressed markedly, with some residual changes. The movement disorders also improved in all patients. CONCLUSION: A syndrome associated with acute bilateral basal ganglia lesions in diabetic uraemic patients is rare, with reversible changes demonstrated by clinical and imaging findings. DWI showed that the bilateral basal ganglia lesions in this syndrome were primarily vasogenic in origin, although there were small foci of cytotoxic oedema within the lesions. PMID- 17922121 TI - Focal laminar cortical infarcts following aneurysmal subarachnoid haemorrhage. AB - INTRODUCTION: The aim of this prospective study was to analyse small band-like cortical infarcts after subarachnoid haemorrhage (SAH) using magnetic resonance imaging (MRI) with reference to additional digital subtraction angiography (DSA). METHODS: In a 5-year period between January 2002 and January 2007 10 out of 188 patients with aneurysmal SAH were evaluated (one patient Hunt and Hess grade I, one patient grade II, four patients grade III, two patients grade IV, and two patients grade V). The imaging protocol included serially performed MRI with diffusion- and perfusion-weighted images (DWI/PWI) at three time points after aneurysm treatment, and cerebral vasospasm (CVS) was analysed on follow-up DSA on day 7+/-3 after SAH. RESULTS: The lesions were located in the frontal lobe (n=10), in the insular cortex (n=3) and in the parietal lobe (n=1). The band-like infarcts occurred after a mean time interval of 5.8 days (range 3-10 days) and showed unexceptional adjacent thick sulcal clots. Seven out of ten patients with cortical infarcts had no or mild CVS, and in the remaining three patients DSA disclosed moderate (n=2) or severe (n=1) CVS. CONCLUSION: The infarct pattern after aneurysmal SAH includes cortical band-like lesions. In contrast to territorial infarcts or lacunar infarcts in the white matter which develop as a result of moderate or severe proximal and/or distal vasospasm visible on angiography, the cortical band-like lesions adjacent to sulcal clots may also develop without evidence of macroscopic vasospasm, implying a vasospastic reaction of the most distal superficial and intraparenchymal vessels. PMID- 17922124 TI - Report on the Fifth Annual Meeting of the Association for Immunotherapy of Cancer (CIMT) April 12-14, 2007 in Wurzburg, Germany. PMID- 17922122 TI - Variations in 123I-metaiodobenzylguanidine (MIBG) late heart mediastinal ratios in chronic heart failure: a need for standardisation and validation. AB - BACKGROUND: There is lack of validation and standardisation of acquisition parameters for myocardial (123)I-metaiodobenzylguanidine (MIBG). This lack of standardisation hampers large scale implementation of (123)I-MIBG parameters in the evaluation of patients with chronic heart failure (CHF). METHODS: In a retrospective multi-centre study (123)I-MIBG planar scintigrams obtained on 290 CHF patients (82% male; 58% dilated cardiomyopathy; New York Heart Association [NYHA classification] > I) were reanalysed to determine the late heart-to mediastinum ratio (H/M). RESULTS: There was a large variation in acquisition parameters. Multivariate forward stepwise regression showed that a significant proportion (31%, p < 0.001) of the variation in late H/M could be explained by a model containing patient-related variables and acquisition parameters. Left ventricular ejection fraction (p < 0.001), type of collimation (p < 0.001), acquisition duration (p = 0.001), NYHA class (p = 0.028) and age (p = 0.034) were independent predictors of late H/M. CONCLUSIONS: Acquisitions parameters are independent contributors to the variation of semi-quantitative measurements of cardiac (123)I-MIBG uptake. Improved standardisation of cardiac (123)I-MIBG imaging parameters would contribute to increased clinical applicability for this procedure. PMID- 17922123 TI - Lung scintigraphy with nonspecific human immunoglobulin G ((99m)Tc-HIG) in the evaluation of pulmonary involvement in connective tissue diseases: correlation with pulmonary function tests (PFTs) and high-resolution computed tomography (HRCT). AB - PURPOSE: In patients with connective tissue diseases (CTD), the early detection and evaluation of the severity of the pulmonary involvement is mandatory. High resolution computed tomography (HRCT) and pulmonary function tests (PFTs) are considered to be valuable noninvasive diagnostic modalities. Radiopharmaceuticals have also been used for this purpose. Our aim was the evaluation of technetium labeled human polyclonal immunoglobulin G (HIG) lung scintigraphy in the early detection and assessment of the severity of the pulmonary involvement in CTD patients. METHODS: Fifty-two nonsmoking CTD patients were studied by PFTs, HRCT, and HIG. According to PFTs, patients were divided in group A (impaired PFTs abnormal pulmonary function) and group B (normal pulmonary function). Semiquantitative analysis was done on HIG and HRCT and corresponding scores were obtained. RESULTS: Significant difference was found between HIG scores in the two groups (0.6 +/- 0.07 vs 0.51 +/- 0.08, P < 0.001). There was a statistically significant negative correlation between HIG scores and PFTs results and a positive correlation between HIG and HRCT scores. HIG demonstrated similar clinical performance to HRCT. At the best cut-off levels of their score (0.56 and 7, respectively), HIG had a superior sensitivity (77.5 vs 57.5%) with lower specificity (75 vs 91.7%). The combination of the two methods increased the sensitivity of abnormal findings at the expense of specificity. CONCLUSIONS: HIG scintigraphy can be used in the early detection and evaluation of the severity of the pulmonary involvement in CTD, whereas, when used in combination with HRCT, the detection of affected patients can be further improved. PMID- 17922125 TI - Erufosine, a novel alkylphosphocholine, in acute myeloid leukemia: single activity and combination with other antileukemic drugs. AB - PURPOSE: Alkylphosphocholines represent a new class of cytostatic drugs with a novel mode of action. Erufosine (ErPC3), the first compound of this class that can be administered intravenously, has recently been shown to be active against human tumor and leukemic cell lines. METHODS: In order to evaluate the antileukemic potential of ErPC3 in acute myeloid leukemia (AML) the lethal concentration 50% (LC 50) was determined using WST-1 assay. For analysis of cell death, staining for Annexin V and activated caspase 3 was performed. An interaction analysis was performed by calculation of combination index and construction of isobolograms. RESULTS: The LC 50 was 7.4 microg/ml after 24 h and 3.2 microg/ml after 72 h in HL 60 cells and 30.1 and 8.6 microg/ml, respectively, in 19 fresh samples from patients with AML. ErPC3 was found to be cytotoxic in HL60 cells with distinct activation of caspase 3. ErPC3 was not cross-resistant with cytarabine, idarubicine and etoposide as shown by the linear relation of respective LC 50s. The latter agents, however, exerted an additive cytotoxicity in combination with ErPC3 as revealed by isobologram analysis and combination index, although results are uneven for idarubicine. CONCLUSION: Based on these data ErPC3 appears as a novel antileukemic candidate drug, which needs to be explored further in the treatment of AML. PMID- 17922126 TI - Extracellular signal-regulated kinase phosphorylation due to menadione-induced arylation mediates growth inhibition of pancreas cancer cells. AB - BACKGROUND: Cytotoxicity of Vitamin K3 (VK3) is indicated to have the same mechanism with oxidative stress (H(2)O(2)). In the present study, we analyzed the differences and/or similarities in the cellular responses to oxidative stress and VK3 to clarify the mechanism of growth inhibition. METHODS: Cell viability was determined by a test method with 3-[4, 5-dimethyl-thiazol]-2, 5-dephenyl tetrazolium bromide (MTT). Expressions of cellular proteins were evaluated by Western blot analysis. RESULTS: The IC50 was calculated to be 47.3 +/- 4.1 microM for VK3 and 2.2 +/- 1.2 microM for H(2)O(2). By Western blot analysis, VK3 or H(2)O(2) was shown to induce rapid phosphorylation of extracellular signal regulated kinase (ERK), and c-Jun N-terminal kinases (JNKs). H(2)O(2)-induced phosphorylation of ERK and JNK was almost complete inhibited by more than 100-muM genistein. VK3-induced JNK phosphorylation was blocked by 100-microM genistein, but ERK phosphorylation was not inhibited completely even if 400-microM genistein was used. H(2)O(2)-induced inhibition of cell proliferation was completely blocked by 400-microM genistein, but the VK3 effect was reduced 72.8 +/- 5.4% by the same concentration of genistein. H(2)O(2)-induced JNK phosphorylation and ERK phosphorylation were inhibited by staurosporine, protein kinase C (PKC) inhibitor. VK3-induced JNK phosphorylation was also blocked, but ERK phosphorylation was not affected. Staurosporine had no effect on VK3- or H(2)O(2) induced growth inhibition. Treatment with a non-thiol antioxidant agent, catalase, completely abrogated H(2)O(2)-induced JNK and ERK phosphorylation, but a thiol antioxidant, L: -cystein, had no effect on phosphorylation of them. The VK3-induced JNK phosphorylation was inhibited by catalase, but not L: -cystein. But ERK phosphorylation was not inhibited by catalase and was abrogated completely by the thiol antioxidant. Catalase, but not L: -cystein, blocked H(2)O(2)-induced growth inhibition, and L: -cystein, but not catalase, blocked VK3-induced effects on cell proliferation completely. CONCLUSION: VK3-induced ERK phosphorylation occurs by a different mechanism from oxidative stress, and it might have an important role to induce growth inhibition. PMID- 17922127 TI - Recombinant erythropoietin differently affects proliferation of mesothelioma cells but not sensitivity to cisplatin and pemetrexed. AB - The combination of cisplatin and pemetrexed represents the newly established standard of care for patients with unresectable malignant mesothelioma (MM). However, this chemotherapy regimen appears to be associated with an increased prevalence of higher grade anemia as compared to treatment with cisplatin alone. Human recombinant erythropoietin (rHuEpo) is currently used for the treatment of anemia in cancer patients. Still, following the finding that the erythropoietin receptor (EpoR) is expressed by several tumor cells types and after the trials reporting that the recombinant cytokine can adversely affect tumor progression and patient survival, the clinical safety of rHuEpo administration to neoplastic patients has recently been questioned. The observation that the expression of EpoR, variably associated with the expression of the cognate ligand, is a common feature of MM cells prompted us to investigate whether treatment with rHuEpo could elicit proliferative and cytoprotective signals in EpoR-positive MM cell lines. Biochemical responsiveness of MM cells to rHuEpo was demonstrated by the time-course activation of both ERK1/2 and AKT following treatment with the recombinant cytokine. A moderately increased mitogenic activity was observed in two out of five MM cell lines treated with pharmacologically relevant concentrations of rHuEpo. On the other hand, the recombinant cytokine, administered either before or after cisplatin and pemetrexed, failed to interfere with the cytotoxic effects exerted by the chemotherapeutic drugs on the five MM cell lines. According to the presented findings, rHuEpo appears to have an overall limited impact on cell growth and no effect on MM sensitivity to chemotherapy. PMID- 17922129 TI - Suture lateralization of vocal cord treating paradoxical vocal cord movement: a case report. AB - Paradoxical vocal cord movement (PVCM) causes inspiratory stridor and extra thoracic airway obstruction through inappropriate adduction of the vocal cords during the respiratory cycle. We report on a patient with severe PVCM necessitating tracheostomy insertion. Using the technique of endoscopic suture lateralization of a vocal cord, we succeeded in decannulation 22 years after tracheostomy. PVCM of this severity is rare and its management remains unclear. Successful treatment in this manner has not been documented previously. A brief review of current management options for this condition is included. PMID- 17922128 TI - Induction of the members of Notch pathway in superficial basal cell carcinomas treated with imiquimod. AB - Basal cell carcinoma of the skin (BCC) is the most common skin tumor in Caucasians worldwide. Different therapeutic options are available to treat BCC, including topical immunotherapy. Imiquimod is topical Toll-like receptor 7 agonist that activates anti-tumor immune response and has been recently approved for the treatment of superficial BCC (sBCC). We sought to investigate the influence of imiquimod treatment on the members of the Notch signaling pathway, whose activity is known to be decreased in BCCs. Six patients with sBCC were evaluated for Notch1, Jagged1 and Delta1 expression before (pre-treatment) and after the beginning of the topical treatment (post-treatment) with imiquimod using real-time PCR and immunohistochemistry. We show selective transcriptional up-regulation of Notch pathway members (Notch1, Jagged1 and Delta1) in tumor cells of the sBCC post-treatment. Furthermore, we demonstrate minor increase of Notch1 protein expression on infiltrating cells as well as strong increase in Jagged1 protein expression in regressing sBCC tumors post-treatment. In this way, imiquimod may act as a stimulator of the Notch pathway in sBCC tumor cells by up regulating protein expression of the Notch ligand, Jagged1. Via induction of Notch signaling imiquimod may exert tumor suppressor function, which together with its proinflammatory properties results in tumor regression. PMID- 17922131 TI - Laryngeal involvement in patients with active pulmonary tuberculosis. AB - The aim of this study was to determine the incidence of laryngeal tuberculosis (LT) among patients with active pulmonary tuberculosis. A total of 319 patients under treatment for pulmonary tuberculosis were subjected to laryngoscopy. Five patients (1.5%) with LT were identified. Odynophagia was the most common complaint, followed by alteration in voice. The larynx returned to its normal appearance in 3-8 months (average 18 weeks) by antituberculous medication. Physicians dealing with pulmonary tuberculosis should keep in mind that symptoms of laryngeal involvement may be minor, and laryngoscopy should always be performed when laryngeal involvement is suspected in order to isolate highly infectious patients. Response to antituberculous medication is usually late in LT and diagnosis by "wait and watch" policy will cause a significant delay in the diagnosis of a possible larynx carcinoma. PMID- 17922132 TI - An animal study on cartilage healing using auricular cartilage as a model. AB - We investigated the effect of different surgical procedures on cartilage healing, using auricular cartilage as a model, which would be useful to create a rationale for septal cartilage surgery. Different kinds of manipulations were performed on the auricular cartilage of six female New Zealand white rabbits. Histopathological investigations were performed under light microscopy 4 months postoperatively. The autologous cartilage grafts survived well under the forehead skin regardless of the presence of the perichondrium. The response of perichondrium to either incomplete or complete trauma was not only new cartilage formation but also ossification. When incomplete incisions were made on the non perichondrial side, new cartilage formation was stimulated whereas ossification was induced when there was perichondrium on the cartilage. If the cartilage with perichondrium was sliced into small pieces and planted back in its original place, many ossification areas occurred. The crushed cartilage was usually absorbed but sometimes replaced by bony plates. The traumatized cartilage with perichondrium undergoes ossification. This finding may be important clinically in that surgeons should not traumatize septal cartilage with perichondrium and work under the perichondrium. This ossification of traumatized cartilage may later result in thickening of the septal cartilage which we sometimes face in revision surgery. PMID- 17922130 TI - A randomised controlled trial comparing Triadcortyl with 10% glycerine-ichthammol in the initial treatment of severe acute otitis externa. AB - Acute otitis externa is a common clinical condition accounting for a large proportion of patients attending the otolaryngology department, although milder cases are often managed in primary care. Treatment of the most severe forms of otitis externa involves aural toilet, followed by the application of a topical preparation, commonly in the form of an ear canal dressing. A prospective single blind randomized controlled trial was performed to compare the efficacy of 10% glycerine-ichthammol (GI) solution and Triadcortyl (TAC) ointment, both applied as ear canal dressings, in the initial management of severe acute otitis externa. A total of 64 patients were studied. Both treatment modalities were proven efficacious in the treatment of severe acute otitis externa. Although there was a statistically significant improvement of pain parameters in the TAC group, we found no significant differences in clinical findings between the two groups. Therefore, it is recommended that GI dressing can be used instead of an antibiotic dressing as an initial treatment of severe acute otitis externa on the basis of cost, avoidance of resistance and toxicity. PMID- 17922133 TI - Absorbed fractions for electrons and beta particles in sensitive regions of human respiratory tract. AB - The absorbed fractions (AF) of electrons in sensitive layers of human respiratory tract were calculated in this paper. For that purpose the source code for simulation package PENELOPE, based on Monte Carlo method, was developed. The human respiratory tract was modeled according to ICRP66 publication, where AF of electrons was calculated using EGS4 simulation software. Some approximations used in ICRP66 were corrected in this work, and new values of AF for radon progeny are given. Minimal energy (EABS) that electron can have during transport through material is 1 keV in ICRP66, while it is set as low as 100 eV in the presented work. Lowering value of EABS gives more accurate results for AF when initial energy of electrons is below 50 keV. To represent tissue, water is used in ICRP66, while in this work epithelia tissue is used. PMID- 17922134 TI - Single lymphocytes from two healthy individuals with mitochondrial point heteroplasmy are mainly homoplasmic. AB - The nature of mitochondrial DNA heteroplasmy is still unclear. It could either be caused by two mitochondrial DNA (mtDNA) haplotypes coexisting within a single cell or by an admixture of homoplasmic cells, each of which contains only one type of mtDNA molecule. To address this question, single lymphocytes were separated by flow cytometry assisted cell sorting and analyzed by cycle sequencing or minisequencing. To attain the required PCR sensitivity, the reactions were carried out on the surface of chemically structured glass slides in a reaction volume of 1-2 microl. In this study, blood samples from two healthy donors showing mitochondrial point heteroplasmy in direct sequencing (195Y and 234R, respectively) were analyzed. Nearly 96% of single lymphocytes tested were found to be in a homoplasmic state, but heteroplasmic cells were also detected. These results suggest that mitochondrial point heteroplasmy in blood may well be mainly due to the mixture of homoplasmic cells. PMID- 17922135 TI - Chondrogenesis, osteogenesis and adipogenesis of canine mesenchymal stem cells: a biochemical, morphological and ultrastructural study. AB - Musculoskeletal diseases with osteochondrotic articular cartilage defects, such as osteoarthritis, are an increasing problem for humans and companion animals which necessitates the development of novel and improved therapeutic strategies. Canine mesenchymal stem cells (cMSCs) offer significant promise as a multipotent source for cell-based therapies and could form the basis for the differentiation and cultivation of tissue grafts to replace damaged tissue. However, no comprehensive analysis has been undertaken to characterize the ultrastructure of in vitro differentiated cMSCs. The main goal of this paper was to focus on cMSCs and to analyse their differentiation capacity. To achieve this aim, bone marrow cMSCs from three canine patients were isolated, expanded in monolayer culture and characterized with respect to their ability for osteogenic, adipogenic and chondrogenic differentiation capacities. cMSCs showed proliferative potential and were capable of osteogenic, adipogenic and chondrogenic differentiation. cMSCs treated with the osteogenic induction medium differentiated into osteoblasts, produced typical bone matrix components, beta1-integrins and upregulated the osteogenic specific transcription factor Cbfa-1. cMSCs treated with the adipogenic induction medium showed typical adipocyte morphology, produced adiponectin, collagen type I and beta1-integrins, and upregulated the adipogenic specific transcription factor PPAR-gamma. cMSCs treated with the chondrogenic induction medium exhibited a round to oval shape, produced a cartilage-specific extracellular matrix, beta1-integrins and upregulated the chondrogenic specific transcription factor Sox9. These results demonstrate, at the biochemical, morphological and ultrastructural levels, the multipotency of cMSCs and thus highlight their potential therapeutic value for cell-based tissue engineering. PMID- 17922136 TI - Acute effects of vibration on thermal perception thresholds. AB - OBJECTIVE: This study focuses on the acute effects of vibration and how vibrations influence the measures of the thermal perception thresholds during different vibration magnitudes, frequencies, and durations. METHODS: The fingers of ten healthy subjects, five males and five females, were exposed to vibration under 16 conditions with a combination of different frequency, intensity and exposure time. The vibration frequency was 31.5 and 125 Hz and exposure lasted between 2 and 16 min. The energy-equivalent frequency weighted acceleration, according to ISO 5349-1, for the experimental time of 16 min was 2.5 or 5.0 m/s(2) (r.m.s.), corresponding to a 8-h equivalent acceleration, A(8) of 0.46 and 0.92 m/s(2), respectively. A measure of the thermal perception of cold and warmth was conducted before the different exposures to vibration. Immediately after the vibration exposure the acute effect was measured continuously on the exposed index finger for the first 75 s, followed by 30 s of measures at every minute for a maximum of 10 min. If the subject's thermal thresholds had not recovered, the measures continued for a maximum of 30 min with measurements taken every 5 min. RESULTS: For all experimental conditions and 30 s after exposure, the mean changes of the thresholds compared with the pre-test were found to be 0.05 and 0.67 degrees C for the warmth and cold thresholds, respectively. The effect of the vibration exposure was only significant on the cold threshold and only for the first minute after exposure when the threshold was decreased. The warmth threshold was not significantly affected at all. The frequency and the exposure time of the vibration stimuli had no significant influence on the perception thresholds for the sensation of cold or warmth. Increased equivalent frequency weighted acceleration resulted in a significant decrease of the subjects' cold threshold, not the warmth. The thresholds were unaffected when changes in the vibration magnitude were expressed as the frequency weighted acceleration or the unweighted acceleration. CONCLUSION: When testing for the thermotactile thresholds, exposure to vibration on the day of a test might influence the results. Until further knowledge is obtained the previous praxis of 2 h avoidance of vibration exposure before assessment is recommended. PMID- 17922137 TI - Evaluation of gene expression through qRT-PCR in cyclically loaded tendons: an in vivo model. AB - An in vivo rabbit animal model for the tendinopathy, epicondylitis, was used to examine the effects of repetitive load on the expression of various genes associated with matrix remodeling. Following 80 h of cumulative load, tissue from the distal and proximal regions of the flexor digitorum profundus tendon was collected. Quantitative RT-PCR was used to asses mRNA levels of collagenase-1 (MMP-1), stromelysin (MMP-3), vascular endothelial growth factor (VEGF), connective tissue growth factor (CTGF), cyclooxygenase-2 (COX-2), interleukin 1beta (IL-1beta), type III collagen (COL-III) and fibronectin (FBRN). No significant differences in expression levels were found between loaded and unloaded limbs at either region of the tendon. The findings were unexpected as the same model has already demonstrated an increase in the density of cells staining for VEGF and CTGF. Different regulatory mechanisms between mRNA and protein expression or localized changes missed due to homogenization of the tissue samples, may explain the discrepancy in findings. PMID- 17922139 TI - Pain management in neonates for painful procedures. PMID- 17922138 TI - Effect of exercise mode on heart rate variability during steady state exercise. AB - This study examined the effect of exercise mode on geometrical, and time and frequency domain measures of heart rate variability (HRV) during steady-state, moderate intensity exercise of the same HR. Seventeen healthy, active male participants volunteered for this study and completed a treadmill VO2max determination. One week later, cardiorespiratory, perceptual and HRV measures were recorded during seated rest (15 min) and consecutive bouts (15 min) of steady-state exercise at 50 and 65% of maximal HR. Exercise was performed using either upper body (arm ergometer), lower body (cycle) or whole body (treadmill) modes. Separated by 1 week and in a random order, participants undertook the same procedures with the remaining exercise modes. Cardiorespiratory, perceptual and HRV responses were determined during rest and steady-state exercise and analysed by two-way (mode vs. stage) repeated measures ANOVA and post hoc pairwise comparisons. Apart from a reduced respiratory rate during lower body exercise, whole and lower body exercise resulted in similar cardiorespiratory, perceptual and HRV responses. Compared to whole or lower body exercise, upper body exercise resulted in significantly (P < 0.05) greater measures of HRV particularly those within the very low (0-0.04 Hz) and low (0.04-0.15 Hz) frequency bands, greater rating of perceived exertion and less oxygen consumption. Upper body, moderate intensity exercise resulted in greater HRV compared to whole or lower body exercise with further studies necessary to elucidate the mechanisms and clinical implications for this greater HRV. PMID- 17922140 TI - Stat3 up-regulates expression of nicotinamide N-methyltransferase in human cancer cells. AB - PURPOSE: To discover new molecular targets for cancer therapy and diagnosis, we surveyed signal transducers and activators of transcription 3 (Stat3)-regulated genes, because constitutive activation of Stat3 is associated with a wide variety of human malignancies. METHODS: We investigated the Stat3-regulated genes in 293 cells with cDNA microarray analysis and found that Nicotinamide N methyltransferase (NNMT) was induced on stimulation of the cells with leukemia inhibitory factor. We examined the expression of NNMT in several types of cancer cells by real-time quantitative RT-PCR. To examine the role of Stat3, Hep-G2 hepatocellular carcinoma cells were transfected with NNMT promoter-luciferase reporter construct together with conditionally active Stat3 (Stat3ER) or dominant negative Stat3 expression vector and NNMT promoter activity was determined. The expression of NNMT and activated Stat3 in 88 colon cancer tissues and 17 normal colon tissues was examined with immunohistochemical analysis. RESULTS: In Hep-G2 cells and SW480 colon cancer cells, NNMT expression increased on stimulation of the cells with interleukin 6. NNMT promoter activity in Hep-G2 cells was dependent on the activation of Stat3. MDA-MB-468 breast cancer cells and HT29 colon cancer cells expressed constitutively a high level of NNMT. Treatment of these cells with Stat3 siRNA or curcumin, which inhibited Stat3 phosphorylation, resulted in reduction of the NNMT level. We found a correlation between the expression of NNMT and activated Stat3 (P<0.001) in the colon cancer tissues. CONCLUSION: NNMT is a novel Stat3-regulated gene. Its expression is enhanced with the activation of Stat3 in colon cancer tissues. NNMT may be a potential candidate for a tumor marker of various kinds of cancers. PMID- 17922142 TI - The neural stem cell niche. AB - The neural stem cell niche defines a zone in which stem cells are retained after embryonic development for the production of new cells of the nervous system. This continual supply of new neurons and glia then provides the postnatal and adult brain with an added capacity for cellular plasticity, albeit one that is restricted to a few specific zones within the brain. Critical to the maintenance of the stem cell niche are microenvironmental cues and cell-cell interactions that act to balance stem cell quiescence with proliferation and to direct neurogenesis versus gliogenesis lineage decisions. Ultimately, based on the location of the niche, stem cells of the adult brain support regeneration in the dentate gyrus of the hippocampus and the olfactory bulb through neuron replacement. Here, we provide a summary of the current understanding of the organization and control mechanisms of the neural stem cell niche. PMID- 17922143 TI - Ectopic adipogenesis of preconditioned adipose-derived stromal cells in an alginate system. AB - Soft tissue substitutes are increasingly needed in reconstructive and plastic surgery, although difficulties arise when autografts or allografts have to provide long-term satisfactory replacement in soft tissue restoration. Adipose tissue engineering has highlighted the possibility of soft tissue restoration by using regeneration strategies. As few reports exist regarding adipose regeneration with alginate systems mixed with adipose-derived stromal cells (ASCs), the present study has attempted to evaluate the adipogenic differentiation of ASCs isolated from BALB/c mice in vitro and to define the potential of ASCs to form ectopic adipose tissue in an alginate system with subcutaneous implantation in nude mice. After 10 days induction in vitro, prediferentiated ASCs were seeded in an alginate gel system and implanted subcutaneously into the dorsum of nude mice. Adipogenesis of the newly formed tissue at 8 weeks post-implantation was confirmed by oil red O staining, reverse transcription/polymerase chain reaction, and Western blot analysis. This study thus provides evidence that the alginate system has the ability to form ectopic adipose tissue when mixed with predifferentiated ASCs. The results also supply clues for improving adipose tissue engineering and soft tissue defects repair in adults. PMID- 17922141 TI - The presence of carboxypeptidase-M in tumour cells signifies epidermal growth factor receptor expression in lung adenocarcinomas: the coexistence predicts a poor prognosis regardless of EGFR levels. AB - PURPOSE: Carboxypeptidase-M (CPM) is a membrane-bound peptidase that metabolizes peptides, and is present in pneumocytes. CPM hydrolyses the C-terminal arginine of epidermal growth factor (EGF) resulting in des-Arg53-EGF which binds to the EGF receptor (EGFR) with an equal or greater affinity than native EGF. Therefore, this study focused on the possible presence of CPM in human lung adenocarcinomas (ADC) and evaluated the relationship between CPM and EGFR by assessing the impact of expressions on patient clinical outcome. METHODS: This is a retrospective study of 110 patients who underwent resection of the primary tumour (92) or metastatic tissues (18) for treatment or diagnosis. Immunohistochemistry (IHC) for CPM and EGFR was made in serial sections using standard methods. RESULTS: This study demonstrates for the first time that 23.6% of ADCs express carboxypeptidase-M (26/110), mainly in membrane-bound forms. The amounts and the extent of CPM within tumours vary from low levels to obviously overexpressed forms. The immunohistochemical positivity (+) for CPM in ADCs negatively correlated with disease survival. In addition, 80% of CPM+ adenocarcinomas (21/26) showed a coexpression with EGFR suggesting a high prevalence for coexistence. The follow up data indicated a significantly shorter 5-year survival time for patients with CPM+-EGFR+ (double-positive) tumours compared to those harbouring neoplasias negative for both proteins (9.5 vs. 60.4% survivals, P < 0.001). CONCLUSION: The fact that CPM+ ADCs often co-express with EGFR suggests a functional-regulatory link between these proteins which might have therapeutical consequences. The present novel data could lead to improved IHC tests in lung adenocarcinomas for EGFR expression. PMID- 17922145 TI - The contractile segment of the abneural limbus in the gecko cochlea is enriched in vimentin. AB - Previously, we discovered a contractile segment within the cartilaginous abneural limbus of the gecko cochlea, the noncartilaginous abneural limbus (NAL, Ganeshina and Vorobyev, J Comp Neurol 461:539-547, 2003). Here, we demonstrate, by means of SDS-PAGE electrophoresis, the nanoLC-ESI-MSMS technique, immunoblotting, and immunocytochemistry, that the major cytoskeletal protein of the NAL cells is vimentin. Filamentous actin constitutes a minor component of the NAL contractile cell cytoskeleton. Our data indicate that the NAL represents a previously unknown specialization of connective tissue, characterized by the reduction of extracellular matrix and a hypertrophy of the vimentin-based intracellular cytoskeleton. The results are compatible with our hypothesis that the NAL is involved in an adaptation of the cochlear mechanics. PMID- 17922144 TI - Skeletal abnormalities and extra-skeletal ossification in mice with restricted Gsalpha deletion caused by a renin promoter-Cre transgene. AB - We have recently generated a transgenic mouse line (termed hRen-Cre) that expresses Cre-recombinase under the control of a 12.2-kb fragment of the human renin promoter. In the present study, we have crossed hRen-Cre mice with a mouse strain in which exon 1 of the Gnas gene is flanked by loxP sites. Gnas encodes the alpha-subunit of the stimulatory G protein (Gs alpha). Our aim has been to generate a mouse model with locally restricted inactivation of Gs alpha to extend studies of the role of Gs alpha function in vivo. Mice with local Cre-mediated inactivation of Gs alpha (rCre-Gs alpha) are viable and fertile. Their most obvious phenotype consists of marked skeletal malformations of the forelimbs in which computer-tomography scans reveal shortened and fused extremity bones. Extraskeletal ossifications occur in the subcutis and in skeletal muscles associated with the affected long bones. Plasma calcium, phosphate and parathyroid hormone are normal. Skin histology has demonstrated diffuse mineralization and ossification associated with the basal cells of hair follicles. This phenotype in part resembles syndromes in humans associated with loss-of-function of Gs alpha, such as Albright hereditary osteodystrophy and progressive osseous heteroplasia. The renal phenotype of rCre-Gs alpha mice is inconspicuous. Plasma renin concentration, ambient urine osmolarity, and the glomerular filtration rate of rCre-Gs alpha mice do not differ from controls. The absence of measurable functional changes in the renin-angiotensin system indicates insufficient Cre expression in juxtaglomerular granular cells in this strain of mice. Nevertheless, the present report reaffirms the importance of Gs alpha signaling for bone development and the suppression of ectopic ossification. PMID- 17922146 TI - Detecting small environmental differences: risk-response curves for predator induced behavior and morphology. AB - Most organisms possess traits that are sensitive to changes in the environment (i.e., plastic traits) which results in the expression of environmentally induced polymorphisms. While most phenotypically plastic traits have traditionally been treated as threshold switches between induced and uninduced states, there is growing evidence that many traits can respond in a continuous fashion. In this experiment we exposed larval anurans (wood frog tadpoles, Rana sylvatica) to an increasing gradient of predation risk to determine how organisms respond to small environmental changes. We manipulated predation risk in two ways: by altering the amount of prey consumed by a constant number of predators (Dytiscus sp.) and by altering the number of predators that consume a constant amount of prey. We then quantified the expression of predator-induced behavior, morphology, and mass to determine the level of risk that induced each trait, the level of risk that induced the maximal phenotypic response for each trait, whether the different traits exhibited a plateauing response, and whether increasing risk via increasing predator number or via increasing prey consumption induced similar phenotypic changes. We found that all of the traits exhibited fine-tuned, graded responses and most of them exhibited a plateauing response with increased predation risk, suggesting either a limit to plasticity or the reflection of high costs of the defensive phenotype. For many traits, a large proportion of the maximum induction occurred at low levels of risk, suggesting that the chemical cues of predation are effective at extremely low concentrations. In contrast to earlier work, we found that behavioral and morphological responses to increased predator number were simply a response to increased total prey consumption. These results have important implications for models of plasticity evolution, models of optimal phenotypic design, expectations for how organisms respond to fine-grained changes (i.e., within generation) in their environment, and impacts on ecological communities via trait-mediated indirect effects. PMID- 17922147 TI - Horseshoe kidney malformation in Turner syndrome is not associated with HNF-1beta gene mutations. AB - Mutations in hepatocyte nuclear factor-1beta (HNF-1beta) gene cause a subtype of maturity-onset diabetes of the young (MODY5), whose clinical features are pancreatic beta-cell dysfunction, renal malformations, and in some females, internal genital malformations. Recently, we reported the first case of MODY5 and horseshoe kidney. The patient was the only male in a three-generation family with five affected females carrying renal cysts or dysplastic kidney. Diabetes mellitus, horseshoe kidney, and X chromosome monosomy or mosaicism can be observed in Turner syndrome (TS). In particular, diabetes mellitus affects about 50% and horseshoe kidney occurs in approximately 16% of patients. To investigate whether mutations/polymorphisms of HNF-1beta and X monosomy influence horseshoe kidney development, we evaluated HNF-1beta gene sequence in 13 patients with TS and several kidney abnormalities. Analysis of the nine exons including intron exon boundaries of HNF-1beta revealed the presence in two subjects (15%) of a known intronic polymorphism, IV8+48insC. No specific variants were found. We conclude there is no direct relationship between horseshoe kidney in TS and mutation or polymorphism of HNF-1beta gene, but we speculate that target gene(s) of HNF-1beta, likely mapped on the X chromosome, is/are responsible of the horseshoe kidney formation in TS. PMID- 17922148 TI - Significant correlation between cerebrospinal fluid nitric oxide concentrations and neurologic prognosis in incomplete cervical cord injury (N. Hosaka et al.). PMID- 17922149 TI - Disc height reduction in adjacent segments and clinical outcome 10 years after lumbar 360 degrees fusion. AB - Adjacent segment degeneration (ASD) is discussed to impair long-term outcome after lumbar interbody fusion. Nevertheless the amount and origin of degeneration and its clinical relevance remain unclear. Only little data is published studying quantitative disc height reduction (DHR) as indicator for ASD in long-term follow up. Forty patients (23 men, 17 women) (group 1: degenerative disc disease, n = 27; group 2: lytic spondylolisthesis, n = 13) underwent lumbar 360 degrees instrumentation and fusion between 1991 and 1997. Preoperative and follow-up lateral lumbar radiographs were studied. Disc heights of first and second cephalad adjacent segments were measured by Farfan's technique and Hurxthal's technique modified by Pope. Clinical outcome was studied using Oswestry disability index (ODI) and visual analogue scale (VAS). Age, gender, prior surgery, fusion rate and number of fusion levels were investigated as potential factors affecting the outcome. Mean follow-up was 114 (72-161) months. Clinical outcome showed an improvement of 44.6% in ODI and 43.8% in VAS with a tendency towards better results in group 2. Fusion rate was 95%. Disc height of the first cephalad adjacent segment in all patients was reduced by on average 21% (Farfan, P < 0.001) and 19% (Pope, P < 0.001), respectively, and that of the second adjacent level by on average 16% (Farfan, P < 0.001) and 14% (Pope, P < 0.001), respectively. A tendency towards more disc height reduction (DHR) in the degenerative group was observed. Advanced age correlated with advanced DHR (P < or = 0.003, r = 0.5). Multiple level fusion led to a more pronounced DHR than 1 level fusion (P = 0.028). There was a tendency towards more DHR in the first adjacent disc compared to the second. Gender, prior surgery of the fused segment and fusion level did not affect the amount of DHR. There was no correlation between the clinical outcome and DHR. Lumbar fusion is associated with DHR of adjacent discs. This may be induced by additional biomechanical stress, ongoing degeneration affecting the lumbar spine and advancing age. However, clinical outcome is not correlated with adjacent DHR. PMID- 17922150 TI - Analysis of five specific scores for cervical spondylogenic myelopathy. AB - The ability to compare various results that measure clinical deficits and outcome is a necessity for successful worldwide discussion about cervical spondylogenic myelopathy (CSM) and its treatment. There is hardly any information in literature how to value and compare outcome assessed by different scores. In a retrospective study we objectively evaluated the Nurick-score, Japanese-orthopaedic-association score (JOA-Score), Cooper-myelopathy-scale (CMS), Prolo-score and European myelopathy-score (EMS) using the data of 43 patients, all of whom showed clinical and morphological signs of CSM and underwent operative decompression. The scores were assessed pre- and postoperatively. The correlation between the score results, anamnesis, clinical and diagnostic data was investigated. All the scores show a statistically significant correlation and measure postoperative improvement. With exception of the Prolo-score all scores reflect clinical deficits of CSM. The Prolo-score rates the severity of CSM on the state of the economic situation above clinical symptoms. The main differences of the scores are shown in the number of patients showing postoperative improvement, varying between 33% (Nurick-score) and 81% (JOA-score). The recovery-rates, as a measure of the cumulative improvement of all the symptoms, show less variation (23-37%). The differences of the recovery-rate were only statistically significant between JOA-score, Nurick-score and EMS (P < 0.05), whereas all the other scores showed no significant differences. To assess the postoperative successes, the evaluation of the recovery-rate is essential. There is no significant difference in the recovery-rate amongst the majority of the scores, which allows a good comparison of the results from different studies. Nevertheless, it is always important to differentiate the therapy results of CSM published worldwide. PMID- 17922151 TI - Cervical myelopathy caused by soft-tissue mass in diffuse idiopathic skeletal hyperostosis. AB - A rare case of cervical spinal cord compression in diffuse idiopathic skeletal hyperostosis (DISH or Forestier's Disease) caused by a craniocervical mass of soft-tissue is reported. The objective is to describe an uncommon mechanism of spinal cord compression in DISH. Three weeks after a cardiac infarction a 69-year old man slowly developed spastic tetraparesis. Magnetic resonance tomography showed a craniocervical tumor compressing the spinal cord and a massive DISH of the cervical spine. An extended mass of yellowish amorphous material was removed from between the dura, the posterior odontoid process and the posterior aspect of vertebral body C2 reaching to the upper part of C3.The histologic appearance indicated connective tissue and cell-degenerated cartilaginous tissue. There was no inflammatory component and no evidence of neoplasia. No ossification of the posterior longitudinal ligament (OPLL) was found. After removal and craniocervical stabilization the patient's neurologic function improved remarkably. The increase of mechanical stress on the atlantoaxial segment and enhanced proliferation reaction of the connective tissue in DISH are suggested as the underlying pathomechanisms in the formation of this soft-tissue mass. PMID- 17922152 TI - The reliability of the Vernon and Mior neck disability index, and its validity compared with the short form-36 health survey questionnaire. AB - Prospective single cohort study. To evaluate the NDI by comparison with the SF36 health Survey Questionnaire. The NDI is a simple ten-item questionnaire used to assess patients with neck pain. The SF36 measures functional ability, well being and the overall health of patients. It is used as a gold standard in health economics to assess the health utility, gain and economic impact of medical interventions. One hundred and sixty patients with neck pain attending the spinal clinic completed self-assessment questionnaires. A second questionnaire was completed in 34 patients after a period of 1-2 weeks. The internal consistency of the NDI and SF36 was calculated using Cronbach's alpha. The test-retest reliability was assessed using the Bland and Altman method. The concurrent validity of the NDI with respect to the SF-36 was assessed using Pearson correlations. Both questionnaires showed robust internal consistency: Cronbach's alpha for the NDI scale was acceptable (0.864, 95% confidence limits 0.825-0.894) though slightly smaller than that of the SF36. The correlations between each item of the NDI scores and the total NDI score ranged from 0.447 to 0.659, (all with P < 0.001). The test-retest reliability of the NDI was high (intra-class correlation 0.93, 95% confidence limits 0.86-0.97) and comparable with the best values found for SF36. The correlations between NDI and SF36 domains ranged from 0.45 to -0.74 (all with P < 0.001). We have shown that the NDI has good reliability and validity and that it compares well with the SF36 in the spinal surgery out patient setting. PMID- 17922153 TI - The role of the endoscope in the transsphenoidal management of cystic lesions of the sellar region. AB - Cystic mass lesions within the sella turcica are common, and they include cystic pituitary adenomas, craniopharyngiomas, Rathke's cleft cysts, arachnoid cysts, and other entities. Until recently, such lesions were typically removed by a microsurgical transsphenoidal route. Given the increased use of the endoscope in transsphenoidal surgery, we evaluated the potential benefits of this tool in the treatment of such lesions. Between January 1997 and March 2005, 76 consecutive patients with sellar-suprasellar cystic lesions treated in three Neurosurgical Divisions underwent transsphenoidal removal in which the endoscope was used at least during the sellar step of the procedure (endoscope-assisted or fully endoscopic). The series consisted of 26 pituitary macroadenomas, 20 Rathke's cleft cysts, 18 craniopharyngiomas, 10 arachnoid cysts, one craniopharyngioma associated with an adrenocorticotropic hormone-secreting adenoma, and one chordoid glioma. Rigid 4-mm endoscopes (0 degrees , 30 degrees , and/or 45 degrees ) were used, and the advantages and limits of the endoscope during the sellar step of the procedure were recorded. Endoscopic exploration after lesion evacuation was generally easier and of greatest efficacy when the residual cystic cavity was larger as opposed to smaller. The use of angled endoscopes was optimal in larger residual cavities. Early descent of the suprasellar cistern, bleeding inside the residual cyst cavity, and a small sella were the most common causes preventing thorough exploration of the residual cavity after its evacuation. In no cases did the endoscope cause injury during the sellar cavity exploration. Endoscopic exploration of the sellar cavity during transsphenoidal surgery offers both general and specific advantages in the treatment of a variety of different cystic sellar lesions. Its routine use during transsphenoidal surgery for such lesions is recommended to achieve maximal and safe tumor removal. PMID- 17922154 TI - Fourier transformed steady-state flash evoked potentials for continuous monitoring of visual pathway function. AB - Monitoring of somatosensory, motor and auditory pathway function by evoked potentials is routine in surgery placing these pathways at risk. However, visual pathway function remains yet inaccessible to a reliable monitoring. For this study, a method of continuous recordings was developed and tested. Steady-state visual evoked potentials were elicited by flash stimulation at 16 Hz and analysed using discrete Fourier transform. Amplitude and phase of the fundamental response were dynamically averaged and continuously plotted in a trend graph. The method was applied on awake individuals with normal vision and on patients undergoing neurosurgery. In most individuals it was possible to continuously record significant responses. Surprisingly, characteristic time-courses of amplitude and phase were observed in several subjects. These findings were attributed mainly to flicker-adaptation. During anesthesia, amplitude and signal-to-noise ratio were markedly smaller. Signal recognition was facilitated when potentials were recorded with a subdural electrode placed directly at the occipital pole. The anesthetic agent propofol had a major impact on the recordings. PMID- 17922155 TI - Effects of prolonged dark adaptation in patients with retinitis pigmentosa of Bothnia type: an electrophysiological study. AB - Bothnia dystrophy (BD) is a variant of recessive retinitis punctata albescens (RPA), caused by the missense mutation R233W in cellular retinaldehyde-binding protein (CRALBP), which is localized in the retinal pigment epithelium (RPE) and Muller cells of the retina. The purpose of this study was, by examining the electrophysiological responses of the retina, to evaluate the capacity of recovery of the whole retinal area and different cell types induced by extremely prolonged dark adaptation (DA) in BD disease and to gain further understanding of the pathogenesis of BD. Six young patients underwent bilateral full-field ERGs after 24 h of DA in one eye and standard DA in the fellow eye. The results were also compared with the effect of prolonged DA (10 h), previously studied in the same patients. After extremely prolonged DA (24 h) the rod b-wave and the mixed rod-cone a-wave responses reached normal though delayed amplitudes. An increase, up to normal level, in the oscillatory response was found. There was no obvious recovery of the cone response. We conclude that in young BD patients during extremely prolonged DA there is a significant additional capacity of recovery of rod function and also significant gain of activity in the inner retinal layer. A continuous but slow regeneration of rod photopigment seems to occur at least up to 24 h. The visual process in the RPE is retarded and CRALBP acts in this process; also, the Muller cells of the retina seem to be involved. The findings also support an extremely slow synthesis of photopigments and irreversibly disturbed cone function early in BD. PMID- 17922157 TI - Reliability and validity studies of the Turkish version of the Epworth Sleepiness Scale. AB - The Epworth Sleepiness Scale (ESS) is a self-administered eight-item questionnaire that is widely used in English speaking countries for assessment of daytime sleepiness in adults. The aim of this study was to investigate the reliability and validity of the ESS in the Turkish language. The Turkish version of the ESS (ESStr) was applied to 194 healthy controls and 150 consecutive subjects attending the sleep centre with symptoms of sleep-disordered breathing. Test-retest reliability of the ESStr was tested in a separate group of 30 subjects. The ESStr scores of 60 subjects with mild to severe obstructive sleep apnoea (OSA) were compared with the ESStr scores of 60 healthy controls matched for age, gender, and body mass index (BMI). Concurrent validity with the Functional Outcomes of Sleep Questionnaire (FOSQtr) was also assessed in 12 subjects. The questionnaire had a high level of internal consistency as measured by Cronbach's alpha (> or =0.86). The test-retest intraclass correlation coefficient was r = 0.81 (95% confidence interval: 0.64-0.90) (p < 0.001) and Spearman's correlation coefficient was r = 0.80 (p = 0.01). The control group had lower ESStr scores than subjects with sleep-disordered breathing (3.6 +/- 3 vs 12.6 +/- 6, respectively; p < 0.001). Subjects with mild sleep-disordered breathing also had lower scores of the ESStr than those with moderate and severe sleep-disordered breathing (10 +/- 6.2 vs 14 +/- 5. and 10 +/- 6.2 vs 16 +/- 5.4, respectively; both p < 0.05), but there were no significant differences between moderate and severe subjects with sleep apnoea. There were significant correlations between the ESStr and total FOSQtr and its subscales (r = -0.22 to r = -0.92; all p = 0.05). Factor analysis of item scores showed that the ESStr had only one factor. The ESStr is a reliable and valid measure of daytime sleepiness. These features and the simplicity of the ESStr make it a valuable measure for clinical management and research. PMID- 17922156 TI - Circulating nitric oxide (NO), asymmetric dimethylarginine (ADMA), homocysteine, and oxidative status in obstructive sleep apnea-hypopnea syndrome (OSAHS). AB - Obstructive sleep apnea-hypopnea syndrome (OSAHS) with episodic hypoxia reoxygenation is associated with increased cardiovascular morbidity and mortality. Therefore, increased homocysteine, asymmetric dimethylarginine (ADMA), oxidative status, and decreased nitric oxide levels have been implicated as possible mechanisms for development of cardiovascular diseases. We aimed to investigate changes in the levels of these substances in patients with OSAHS in comparison with nonapneic controls. Thirty-four OSAHS patients and 15 healthy controls were included in this study. In the blood samples, oxidative status and nitric oxide levels were measured with spectrophotometric methods. Plasma ADMA and homocysteine levels were determined by using high-performance liquid chromatography with fluorescence detection. Nitric oxide levels were significantly low in OSAHS patients (p < 0.05) and correlated with mean SaO(2) (r = 0.513, p < 0.002) and lowest SaO(2) (r = 0.363, p < 0.03). Oxidative status, ADMA, and homocysteine levels were higher in OSAHS patients, but difference did not reach statistical significance. After dividing patients into moderate (AHI = 5-29) and severe (AHI > or = 30) OSAHS groups, significantly increased homocysteine levels were observed in the severe OSAHS group (p < 0.05). Nitric oxide levels negatively correlated with oxidative status in total OSAHS patients (r = -0.415, p < 0.02) and also in severe OSAHS group (r = -0.641, p < 0.007). Hyperhomocysteinemia and diminished NO production may be causal factors in endothelial dysfunction seen in OSAHS and may explain the association between OSAHS and cardiovascular diseases. These modifiable factors should be monitored in patients suspected of having OSAHS. PMID- 17922158 TI - Evaluation of a new sitting concept designed for prevention of pressure ulcer on the buttock using finite element analysis. AB - Excessive compressive load induces pressure related soft tissue damage, i.e. pressure ulcer (PU), in buttock area in wheelchair users. In solving this problem, our previous study has introduced a concept of Off-Loading sitting, which partially removes the ischial support to reduce pressure under buttocks. However, the effect of this sitting concept has only been evaluated using the interface pressure and tissue perfusion measurements. The objective of this investigation was to evaluate the Off-Loading posture for its ability to reduce internal pressure and stress in deep buttock tissues. This evaluation was performed on a 3D finite element (FE) model which was established and validated in a sitting posture and has realistic material properties and boundary conditions. FE analysis in this study confirmed that the pressure relief provided by Off-Loading posture created profound effect in reducing the mechanical stress within deep tissues. It was concluded that Off-Loading posture may prove beneficial in preventing sitting related PU. PMID- 17922161 TI - Embolization of an insulinoma of the pancreas with trisacryl gelatin microspheres as definitive treatment. AB - Insulinomas are rare, mostly benign neuroendocrine tumors, originating in 99% of cases from the pancreas, that synthesize and secrete insulin, causing symptomatic hypoglycemia. Today the treatment of choice is surgical removal. We present the case of an 84-year-old woman with a symptomatic insulinoma who refused surgery and was treated with arterial embolization using trisacryl gelatin microspheres as definitive treatment. PMID- 17922160 TI - Hepatic arterial embolization and chemoembolization in the management of patients with large-volume liver metastases. AB - The purpose of this study was to assess the role of hepatic arterial embolization (HAE) and chemoembolization (HACE) in patients with large-volume liver metastases. Patients with metastatic neuroendocrine tumors, melanomas, or gastrointestinal stromal tumors (GISTs) with >75% liver involvement who underwent HAE or HACE were included in the study. Radiologic response, progression-free survival (PFS), overall survival (OS), and postprocedure complications were assessed. Sixty patients underwent 123 treatment sessions. Of the 48 patients for whom follow-up imaging was available, partial response was seen in 12 (25%) patients, minimal response in 6 (12%), stable disease in 22 (46%), and progressive disease in 8 (17%). Median OS and PFS were 9.3 and 4.9 months, respectively. Treatment resulted in radiologic response or disease stabilization in 82% and symptomatic response in 65% of patients with neuroendocrine tumors. Patients with neuroendocrine tumors had higher response rates (44% vs. 27% and 0%; p = 0.31) and longer PFS (9.2 vs. 2.0 and 2.3 months; p < 0.0001) and OS (17.9 vs. 2.4 and 2.3 months; p < 0.0001) compared to patients with melanomas and GISTs. Major complications occurred in 21 patients after 23 (19%) of the 123 sessions. Nine of the 12 patients who developed major complications resulting in death had additional risk factors--carcinoid heart disease, sepsis, rapidly worsening performance status, or anasarca. In conclusion, in patients with neuroendocrine tumors with >75% liver involvement, HAE/HACE resulted in symptom palliation and radiologic response or disease stabilization in the majority of patients. Patients with hepatic metastases from melanomas and GISTs, however, did not show any appreciable benefit from this procedure. Patients with massive liver tumor burden, who have additional risk factors, should not be subjected to HAE/HACE because of the high risk of procedure-related mortality. PMID- 17922162 TI - Glucagon-induced vasospasm of hepatic artery branches during visceral angiography. AB - Glucagon is often used in radiology to decrease bowel motility for enhanced imaging, including visceral digital subtraction angiography. We present a case in which branch hepatic artery vasospasm followed the intravenous administration of glucagon during visceral angiography. PMID- 17922163 TI - Endovascular exclusion of visceral artery aneurysms with stent-grafts: technique and long-term follow-up. AB - This paper describes four cases of visceral artery aneurysms (VAAs) successfully treated with endovascular stent-grafts and discusses the endovascular approach to VAAs and the long-term results. Four balloon expandable stent-grafts were used to treat three splenic artery aneurysms and one bleeding common hepatic artery pseudoaneurysm. The percutaneous access site and the materials were chosen on the basis of CT angiography findings. In all cases the aneurysms were successfully excluded. In one case a splenic infarction occurred, with nonrelevant clinical findings. At 16- to 24-month follow-up three patients had patent stents and complete exclusion and shrinkage of the aneurysms. One patient died due to pancreatitis and sepsis, 16 days after successful stenting and exclusion of a bleeding pseudoaneurysm. We conclude that endovascular treatment using covered stent-grafts is a valid therapeutic option for VAAs. Multislice CT preoperative study helps in planning stent-graft positioning. PMID- 17922164 TI - Characterization of a SEPT9 interacting protein, SEPT14, a novel testis-specific septin. AB - Septins are a highly conserved family of GTP-binding cytoskeletal proteins implicated in multiple cellular functions, including membrane transport, apoptosis, cell polarity, cell cycle regulation, cytokinesis, and oncogenesis. Here we describe the characterization of a novel interacting partner of the septin family, initially cloned from a human testis expression library following yeast two-hybrid isolation to identify SEPT9 binding partners. Upon further genomic characterization and bioinformatics analyses it was determined that this novel septin-interacting partner was also a new member of the mammalian septin family, named SEPT14. SEPT14 maps to 7p11.2 in humans and includes a conserved GTPase domain and a predicted carboxy-terminus coiled-coil domain characteristic of other septins. Three potential translational start methionines were identified by 5' RACE-PCR encoding proteins of 432-, 427-, and 425-residue peptides, respectively. SEPT14 shares closest homology to SEPT10, a human dendritic septin, and limited homology to SEPT9 isoforms. SEPT14 colocalized with SEPT9 when coexpressed in cell lines, and epitope-tagged forms of these proteins coimmunoprecipitated. Moreover, SEPT14 was coimmunoprecipitated from rat testes using SEPT9 antibodies, and yeast two-hybrid analysis suggested SEPT14 interactions with nine additional septins. Multitissue Northern blotting showed testis-specific expression of a single 5.0-kb SEPT14 transcript. RT-PCR analysis revealed that SEPT14 was not detectable in normal or cancerous ovarian, breast, prostate, bladder, or kidney cell lines and was only faintly detected in fetal liver, tonsil, and thymus samples. Interestingly, SEPT14 was expressed in testis but not testicular cancer cell lines by RT-PCR, suggesting that further investigation of SEPT14 as a testis-specific tumor suppressor is necessary. PMID- 17922165 TI - Self-confidence in and perceived utility of the physical examination: a comparison of medical students, residents, and faculty internists. AB - BACKGROUND AND OBJECTIVES: Little is known about the differences in attitudes of medical students, Internal Medicine residents, and faculty Internists toward the physical examination. We sought to investigate these groups' self-confidence in and perceived utility of physical examination skills. DESIGN AND PARTICIPANTS: Cross-sectional survey of third- and fourth-year medical students, Internal Medicine residents, and faculty Internists at an academic teaching hospital. MEASUREMENTS: Using a 5-point Likert-type scale, respondents indicated their self confidence in overall physical examination skill, as well as their ability to perform 14 individual skills, and how useful they felt the overall physical examination, and each skill, to be for yielding clinically important information. RESULTS: The response rate was 80% (302/376). The skills with overall mean self confidence ratings less than "neutral" were interpreting a diastolic murmur (2.9), detecting a thyroid nodule (2.8), and the nondilated fundoscopic examination using an ophthalmoscope to assess retinal vasculature (2.5). No skills had a mean utility rating less than neutral. The skills with the greatest numerical differences between mean self-confidence and perceived utility were distinguishing between a mole and melanoma (1.5), detecting a thyroid nodule (1.4), and interpreting a diastolic murmur (1.3). Regarding overall self confidence, third-year students' ratings (3.3) were similar to those of first year residents (3.4; p = .95) but less than those of fourth-year students (3.8; p = .002), upper-level residents (3.7; p = .01), and faculty Internists (3.9; p < .001). CONCLUSIONS: Self-confidence in the physical exam does not necessarily increase at each stage of training. The differences found between self-confidence and perceived utility for a number of skills suggest important areas for educational interventions. PMID- 17922166 TI - Frustrated and confused: the American public rates its cancer-related information seeking experiences. AB - BACKGROUND: Ensuring access to high-quality cancer-related information is important for the success of cancer prevention and control efforts. OBJECTIVE: We conducted a population-based assessment of the barriers faced by people searching for cancer information. DESIGN: Cross-sectional data from the National Cancer Institute's 2003 Health Information National Trends Survey. PARTICIPANTS: A nationally representative sample of individuals in the USA (n = 6,369). MEASUREMENTS: We assessed whether respondents had ever sought cancer-related information and examined ratings of their information-seeking experiences and beliefs regarding causes of cancer and its prevention. Linear and logistic regression models were estimated to determine predictors of negative experiences and associations between experiences and cancer beliefs. RESULTS: Nearly one half (44.9%) of Americans had searched for cancer information. Many reported negative experiences, including the search process requiring a lot of effort (47.7%), expressing frustration (41.3%), and concerns about the quality of the information found (57.7%). Respondents lacking health insurance or a high school education experienced the greatest difficulty. Compared to those reporting the most positive experiences, information seekers reporting more negative experiences were more likely to report that almost everything caused cancer [odds ratio (OR) 2.0, 95% confidence interval (CI) 1.5-2.6], that not much can be done to prevent cancer (OR 2.7, 95% CI 1.9-3.8), and that it is hard to know which cancer prevention recommendations to follow (OR 3.2, 95% CI 2.3-4.5). CONCLUSIONS: While a significant proportion of the American public searches for cancer information, suboptimal experiences are common. Facilitation of information seeking will be critical for promoting informed decision making in cancer prevention and control. PMID- 17922167 TI - Effectiveness of a lifestyle intervention on metabolic syndrome. A randomized controlled trial. AB - BACKGROUND: Intensive lifestyle intervention significantly reduces the progression to diabetes in high-risk individuals. OBJECTIVE: It is not known whether a program of moderate intervention might effectively reduce metabolic abnormalities in the general population. DESIGN: Two-arm randomized controlled 1 year trial. PATIENTS: Three hundred and thirty-five patients participated from a dysmetabolic population-based cohort of 375 adults aged 45-64 years in northwestern Italy. MEASUREMENTS: We compared the effectiveness of a general recommendation-based program of lifestyle intervention carried out by trained professionals versus standard unstructured information given by family physicians at reducing the prevalence of multiple metabolic and inflammatory abnormalities. RESULTS: At baseline, clinical/anthropometric/laboratory and lifestyle characteristics of the intervention (n = 169) and control (n = 166) groups were not significantly different. The former significantly reduced total/saturated fat intake and increased polyunsaturated fat/fiber intake and exercise level compared to the controls. Weight, waist circumference, high-sensitivity C-reactive protein, and most of the metabolic syndrome components decreased in the intervention group and increased in the controls after 12 months. Lifestyle intervention significantly reduced metabolic syndrome (odds ratio [OR] = 0.28; 95% CI 0.18-0.44), with a 31% (21-41) absolute risk reduction, corresponding to 3.2 (2-5) patients needing to be treated to prevent 1 case after 12 months. The intervention significantly reduced the prevalence of central obesity (OR = 0.33; 0.20-0.56), and hypertriglyceridemia (OR = 0.48; 0.31-0.75) and the incidence of diabetes (OR = 0.23; 0.06-0.85). CONCLUSION: A lifestyle intervention based on general recommendations was effective in reducing multiple metabolic/inflammatory abnormalities. The usual care by family physicians was ineffective at modifying progressive metabolic deterioration in high-risk individuals. PMID- 17922168 TI - Effectiveness of a 1-year resident training program in clinical research: a controlled before-and-after study. AB - BACKGROUND: To increase the number of clinician scientists and to improve research skills, a number of clinical research training programs have been recently established. However, controlled studies assessing their effectiveness are lacking. OBJECTIVE: To investigate the effectiveness of a 1-year resident training program in clinical research. DESIGN: Controlled before-and-after study. The training program included a weekly class in clinical research methods, completion of a research project, and mentorship. PARTICIPANTS: Intervention subjects were 15 residents participating in the 1-year training program in clinical research. Control subjects were 22 residents not participating in the training program. MEASUREMENTS AND MAIN RESULTS: Assessments were performed at the beginning and end of the program. Outcomes included methodological research knowledge (multiple-choice progress test), self-assessed research competence, progress on publications and grant applications, and evaluation of the program using quantitative and qualitative methods. RESULTS: Intervention subjects and controls were well matched with respect to research experience (5.1 +/- 2.2 vs 5.6 +/- 5.8 years; p = .69). Methodological knowledge improved significantly more in the intervention group compared to the control group (effect size = 2.5; p < .001). Similarly, self-assessed research competence increased significantly more in the intervention group (effect size = 1.1; p = .01). At the end of the program, significantly more intervention subjects compared to controls were currently writing journal articles (87% vs 36%; p = .003). The intervention subjects evaluated the training program as highly valuable for becoming independent researchers. CONCLUSIONS: A 1-year training program in clinical research can substantially increase research knowledge and productivity. The program design makes it feasible to implement in other academic settings. PMID- 17922169 TI - Somatic symptoms and diseases are more common in women exposed to violence. AB - BACKGROUND: Exposure to violence has been shown to have an impact on somatic health. However, our knowledge about the possible dose-response relationship between frequency of violence exposure and health is still limited. OBJECTIVE: To study the associations between recent and repetitive exposure to violence and presence of somatic symptoms and diseases in women. DESIGN: Cross-sectional, community-based, self-reporting survey. PARTICIPANTS: Two thousand seven hundred thirty women aged 18-40 years (mean age 30.5 years). MEASUREMENTS: The somatic symptom scale derived from the Primary Care Evaluation of Mental Disorders was used to obtain information on the presence of somatization. In addition, we asked about the presence of 11 diseases or organ-specific diseases. Exposure to violence was measured by the Abuse Assessment Screen. RESULTS: Eighteen percent (486/2,730) of women surveyed reported exposure to physical violence. Three percent (94/2,730) had been forced into sexual intercourse as an adult. All somatic symptoms, and several diseases, were significantly more common in women exposed to physical and/or sexual violence as compared to nonexposed women. Women exposed to 3 or more violent episodes in the past 12 months reported a presence of 4.8 somatic symptoms and 1.2 diseases (mean) as compared to 1.8 symptoms and 0.5 diseases in nonexposed women. Women with exposure to both physical and sexual violence reported 6.0 symptoms and 1.5 diseases. The impact of violence on somatic symptoms and diseases remained after controlling for depression and sociodemographic factors. CONCLUSIONS: Violence was associated with the presence of somatic symptoms and diseases. The more a woman is exposed to violence, the higher the number of somatic symptoms and diseases reported is. PMID- 17922170 TI - Real-time EBM: from bed board to keyboard and back. AB - BACKGROUND: To practice Evidence-Based Medicine (EBM), physicians must quickly retrieve evidence to inform medical decisions. Internal Medicine (IM) residents receive little formal education in electronic database searching, and have identified poor searching skills as a barrier to practicing EBM. OBJECTIVE: To design and implement a database searching tutorial for IM residents on inpatient rotations and to evaluate its impact on residents' skill and comfort searching MEDLINE and filtered EBM resources. DESIGN: Randomized controlled trial. Residents randomized to the searching tutorial met for up to 6 1-hour small group sessions to search for answers to questions about current hospitalized patients. PARTICIPANTS: Second- and 3rd-year IM residents. MEASUREMENTS: Residents in both groups completed an Objective Structured Searching Evaluation (OSSE), searching for primary evidence to answer 5 clinical questions. OSSE outcomes were the number of successful searches, search times, and techniques utilized. Participants also completed self-assessment surveys measuring frequency and comfort using EBM databases. RESULTS: During the OSSE, residents who participated in the intervention utilized more searching techniques overall (p < .01) and used PubMed's Clinical Queries more often (p < .001) than control residents. Searching "success" and time per completed search did not differ between groups. Compared with controls, intervention residents reported greater comfort using MEDLINE (p < .05) and the Cochrane Library (p < .05) on post-intervention surveys. The groups did not differ in comfort using ACP Journal Club, or in self-reported frequency of use of any databases. CONCLUSIONS: An inpatient EBM searching tutorial improved searching techniques of IM residents and resulted in increased comfort with MEDLINE and the Cochrane Library, but did not impact overall searching success. PMID- 17922172 TI - Long-term adherence to evidence based secondary prevention therapies after acute myocardial infarction. AB - BACKGROUND: After acute myocardial infarction (AMI), treatment with beta-blockers and angiotensin-converting enzyme inhibitors (ACEI) is widely recognized as crucial to reduce risk of a subsequent AMI. However, many patients fail to consistently remain on these treatments over time, and long-term adherence has not been well described. OBJECTIVE: To examine the duration of treatment with beta-blockers and ACEI within the 24 months after an AMI. DESIGN: A retrospective, observational study using medical and pharmacy claims from a large health plan operating in the Northeastern United States. SUBJECTS: Enrollees with an inpatient claim for AMI who initiated beta-blocker (N = 499) or ACEI (N = 526) therapy. MEASUREMENT: Time from initiation to discontinuation was measured with pharmacy refill records. Associations between therapy discontinuation and potential predictors were estimated using a Cox proportional hazards model. RESULTS: ACEI discontinuation rates were high: 7% stopped within 1 month, 22% at 6 months, 32% at 1 year and 50% at 2 years. Overall discontinuation rates for beta-blockers were similar, but predictors of discontinuation differed for the two treatment types. For beta-blockers, the risk of discontinuation was highest among males and those from low-income neighborhoods; patients with comorbid hypertension and peripheral vascular disease were less likely to discontinue therapy. These factors were not associated with ACEI discontinuation. CONCLUSION: Many patients initiating evidence-based secondary prevention therapies after an AMI fail to consistently remain on these treatments. Adherence is a priority area for development of better-quality measures and quality-improvement interventions. Barriers to beta-blocker adherence for low-income populations need particular attention. PMID- 17922171 TI - Primary care visit length, quality, and satisfaction for standardized patients with depression. AB - BACKGROUND: The contribution of physician and organizational factors to visit length, quality, and satisfaction remains uncertain, in part, because of confounding by patient presentation. OBJECTIVE: To determine associations among visit length, quality, and satisfaction when patient presentation is controlled. DESIGN: A factorial experiment using standardized patients to make primary care visits presenting with either major depression or adjustment disorder, and a musculoskeletal complaint. PARTICIPANTS: One hundred fifty-two primary care physicians, each seeing 2 standardized patients. MEASUREMENTS: Visit length was determined from surreptitiously obtained audiorecordings. Other key measures were derived from physician and standardized patient report. RESULTS: Mean visit length for 294 completed encounters was 22.3 minutes (range = 5.8-72.2, SD = 9.4). Key factors associated with visit length were: physician style (rho = 0.68 and 0.54 after multivariate adjustment), nonprofessional experience with depression (11% longer, 95% CI = 0-23%), practicing within an HMO (26% shorter, 95% CI = 61-90%), and greater practice volume (those working >9 half-day clinic sessions/week had 15% shorter visits than those working fewer than 6, 95% CI = 0 27%, and those seeing >12 patients/half-day had 27% shorter visits than those seeing <10 patients/half-day, 95% CI = 13-39%). Suicidal inquiry (a process-based quality-of-care measure for depression) was not associated with adjusted visit length. Satisfaction was linearly associated with visit length but not with suicide inquiry or follow-up interval. CONCLUSIONS: Despite experimental control for clinical presentation, wide variation in visit length persists, largely reflecting individual physician styles. Visit length is a significant determinant of standardized patient satisfaction. PMID- 17922173 TI - Mucin-producing bile duct tumor of the caudate lobe protruding into the common hepatic duct. AB - Mucin-producing tumor in the bile duct is referred to clinically as mucin producing bile duct tumor (MPBT). Intraductal papillary neoplasm of the biliary tract that resembles an intraductal papillary mucinous neoplasm (IPMN) of the pancreas is a rare category of MPBT and is not well characterized. We, herein, report a case of MPBT of the caudate lobe of the liver that showed papillary growth and communicated with the bile duct of the caudate lobe and protruded into the common hepatic duct. Histologically, MPBT cells showed papillary overgrowth with abundant mucinous secretions, resembling an IPMN of the pancreas. The MPBT cells showed the same immunostaining pattern as that of cells from IPMN of the pancreas. PMID- 17922174 TI - Effects of methylprednisolone and its liver-targeted dextran prodrug on ischemia reperfusion injury in a rat liver transplantation model. AB - PURPOSE: To evaluate the effectiveness of a liver-targeted dextran prodrug (DMP) of methylprednisolone (MP) in cold preservation-warm reperfusion injury associated with liver transplantation. METHODS: The effects of donor pretreatment with single 5 mg/kg doses of MP or DMP on ischemia-reperfusion damage to the liver were studied after 8 or 24 h of cold preservation in both isolated perfused rat liver (IPRL) and syngeneic orthotopic rat liver transplantation (OLT) models. RESULTS: In IPRL studies, donor pretreatment with DMP, and to a lesser degree MP, significantly improved the uptake of hyaluronic acid (HA), a marker of endothelial cell function, following 8 h of cold preservation. However, neither pretreatment was protective after 24 h of preservation. In the OLT model using 24 h-preserved livers, the seven-day survival of untreated grafts was 50%. DMP pretreatment of donors significantly improved graft survival to 100%, whereas MP pretreatment was ineffective. Additionally, only DMP significantly increased the blood glucose concentrations and decreased the plasma concentrations of tumor necrosis factor-alpha after OLT. Other measured markers of liver injury were not affected by either pretreatment. CONCLUSIONS: Selective delivery of methylprednisolone to the liver as a donor pretreatment strategy improves 24-h preserved graft survival in the OLT model. PMID- 17922175 TI - Plasmonic enhancement of single-molecule fluorescence near a silver nanoparticle. AB - In this short paper, we reported the enhanced fluorescence from a single fluorophore bound to a 50nm silver nanoparticle. We found that on average the Cy5 molecules bound to metal nanoparticles are approximately 15-fold brighter than that of free dyes, and that single molecule lifetimes are shorter as compared to free fluorophores. The increased emission rate is primarily the result of local plasmon enhancement. These results demonstrate that the use of fluorophore-metal interactions can increase the brightness and photostability of fluorophores for single molecule detection. PMID- 17922178 TI - The Standardized Field Sobriety Tests: a review of scientific and legal issues. AB - This article details the history and development of the National Highway and Safety Administration's Standardized Field Sobriety Tests. They are reviewed in terms of relevant scientific, psychometric, and legal issues. It is concluded that the research that supports their use is limited, important confounding variables have not been thoroughly studied, reliability is mediocre, and that their developers and prosecution-oriented publications have oversold the tests. Further, case law since their development has severed the tests from their validation data, so that they are not admissible on the criterion for which they were validated (blood alcohol concentration), and admissible for a criterion for which they were not (mental, physical, or driving impairment). Directions for further research are presented. PMID- 17922179 TI - Asperger (1906-1980) and Kanner (1894-1981), the two pioneers of autism. PMID- 17922180 TI - Creation of diversity in the animal virus world by inter-species and intra species recombinations: lessons learned from poultry viruses. AB - The biological diversity within viruses is one of the largest found in all other forms of nature. Many mechanisms contribute to virus diversity and include incorporating genetic material from the host, recombination between viruses belonging to the same or to a different family, and even recombination between viruses normally infecting different hosts. In particular, avian viruses can utilize all three of these mechanisms to generate new viruses. It is well documented that recombinations can occur between Marek's disease virus (MDV), an oncogenic herpesvirus, fowlpox virus (FPV), and various avian retroviruses. In addition, chicken infectious anemia virus (CIAV), a circovirus, was created by several inter-family recombination events, which occurred between plant and animal viruses. The circovirus represents the ancestral creation of a recombination between a plant DNA virus (nanovirus) and a mammalian RNA virus (calicivirus), through a transition of RNA to DNA made by an endogenous mammalian retrovirus. The present review will discuss the current knowledge on recombination events that have occurred between avian herpesviruses and retroviruses following dual infections in vitro and in vivo. In addition, we will discuss recombinations between fowlpox viruses and the avian retrovirus reticuloendotheliosis (REV). Finally, the review will address the creation of CIAV and how it evolved from recombinations between a plant virus and an animal virus. PMID- 17922182 TI - An assessment of HIV/STI vulnerability and related sexual risk-taking in a nationally representative sample of young Croatian adults. AB - Despite the recent increase in the number of HIV infections in Central and Eastern Europe, patterns of sexual behavior have not been extensively researched, particularly among young people. The aim of the present study was to provide a comprehensive assessment of HIV/AIDS-related vulnerability and sexual risk-taking among young adults in Croatia. Data were collected in 2005 using a nationally representative, multi-stage stratified probability sample (n = 1,093) of women and men aged 18-24 years. The focus in this article was on predictors of sexual risk-taking measured by a composite risky sexual behaviors scale. Using hierarchical regression models, we analyzed gendered effects of community, family, peer group, and individual level factors. For both men and women, peer pressure, sensation seeking, personal risk-assessment, behavioral intention, condom use at first sexual intercourse, and sexual victimization were significant predictors of sexual risk-taking behaviors. A number of predictors were gender specific: sexual assertiveness and condom self-efficacy for women and parental monitoring, traditional morality, HIV knowledge, and talking about sex with partner for men. Documenting substantial prevalence of potentially risky sexual behaviors among young people in Croatia, the findings call for prevention and intervention efforts that should focus on individual capacity building for responsible sexual behavior. PMID- 17922181 TI - Co-expressing GP5 and M proteins under different promoters in recombinant modified vaccinia virus ankara (rMVA)-based vaccine vector enhanced the humoral and cellular immune responses of porcine reproductive and respiratory syndrome virus (PRRSV). AB - The porcine reproductive and respiratory syndrome virus (PRRSV) has three major structural proteins which designated as GP5, M, and N. Protein GP5 and M have been considered very important to arouse the humoral and cellular immune responses against PRRSV infection and proposed to be the excellent candidate proteins in the design of PRRS bioengineering vaccine. There were some attempts on expressing GP5 or M in DNA vaccine and adenovirus to arouse humoral and cellular immune responses, but few papers have been reported on that the immune response can be difference because of the expression patterns of GP5 and M proteins in the recombinant virus. In this article, four recombinant viruses that expressed GP5 and M proteins of PRRSV in the modified vaccinia virus ankara (MVA) with different expression patterns were made. In these recombinant virus (rMVAs), GP5 and M proteins were expressed in MVA in the same virus but under the control of two promoters (rMVA-GP5/M), or as a fusion protein under one promoter (rMVA GP5-M), or separately (rMVA-GP5 and rMVA-M). The humoral and cellular immune responses for the four recombinant viruses were evaluated with mouse model. Every mouse was inoculated with 5 x 10(5) TCID50 of the different rMVAs and boosted 3 weeks later. Neutralizing antibody titers for each group were detected with virus neutralization test assay weekly after the primary inoculation for 13 weeks to evaluate the humoral immune response. The production of gamma interferon (IFN gamma), interleukin-2 (IL-2), and interleukin-4 (IL-4) was detected in splenocytes of rMVA-inoculated mice at 30, 60, and 90 days post inoculation to evaluate the cellular immune response. Results showed that rMVA-GP5 and rMVA-M cannot induce obvious humoral and cellular immune responses; rMVA-GP5-M inoculated group developed better immune responses than rMVA-GP5 and rMVA-M inoculated groups; however, mice inoculated with rMVA-GP5/M maintained the strongest cellular response against PRRS and consistently enhanced the anti-PRRSV humoral responses. The strategy of co-expressing PRRSV GP5 and M protein in MVA under the control of different promoters might be an attractive method for future PRRSV vaccine design. PMID- 17922184 TI - Inhibition in adults with attention deficit/hyperactivity disorder: event-related potentials in the stop task. AB - The core deficit in Attention Deficit/Hyperactivity Disorder (ADHD) may be a deficiency in executive functions, particularly the processes that are associated with the inhibition of predominant responses. To test this notion in the adult population, healthy undergraduate volunteers and students with ADHD symptoms performed a visual Stop Signal Task (Logan et al. J Exp Psychol: Hum Percept Perform 10:276-291, 1984) while Event-Related brain Potentials were recorded. The two groups did not differ on behavioral measures of performance, but there was a significant difference in the N2-P3 component. These results underline the robustness of an N2-P3 difference between healthy adults and people with ADHD symptoms that have persisted into young adulthood. PMID- 17922183 TI - Dynamic processing of nociception in cortical network in conscious rats: a laser evoked field potential study. AB - (1) Field potential study in conscious rats provides a convenient and effective animal model for pain mechanism and pharmacological research. However, the spatial-temporal character of nociception processing in cortex revealed by field potential technique in conscious rats remains unclear. (2) In the present study, multi-channel field potentials evoked by noxious laser stimulation applied to the hind paw of conscious rats were recorded through 12 chronically implanted skull electrodes. Independent component analysis (ICA) was used to remove possible artifacts and to extract the specific nociception-related component. (3) Two fast sharp responses and one slow blunt response were evoked by noxious laser stimulation. Systemic morphine (5 mg/kg, i.p.) preferentially attenuated the amplitude of the slow blunt response while had no significant effect on the first two sharp responses. ICA revealed that those responses came from activities of contralateral anterior parietal area, medial frontal area and posterior parietal area. A movement artifact was also detected in this study. Partial directed coherence (PDC) analysis showed that there were changes of information flows from medial frontal and posterior parietal area to anterior parietal area after noxious laser stimulation. (4) Characterization of the spatio-temporal responses to noxious laser stimulation may be a valuable model for the study of pain mechanisms and for the assessment of analgesia. PMID- 17922185 TI - Prospective characterization of musculoskeletal symptoms in early stage breast cancer patients treated with aromatase inhibitors. AB - PURPOSE: Aromatase inhibitors (AIs) are increasingly used as adjuvant treatment of postmenopausal women with hormone receptor-positive breast cancer. AIs are commonly associated with musculoskeletal symptoms. The primary objective of this study was to describe the musculoskeletal symptoms that developed in the first 100 subjects enrolled who had at least 6 months follow-up. METHODS: Women with early stage hormone receptor-positive breast cancer were recruited into a multicenter randomized clinical trial to study the pharmacogenomics of two AIs, exemestane, and letrozole. Patients completed the Health Assessment Questionnaire (HAQ) and Visual Analog Scale (VAS) at baseline, 1, 3, 6, and 12 months to assess changes in function and pain, respectively. Patients were referred for evaluation by a rheumatologist if their HAQ and/or VAS scores exceeded a predefined threshold. RESULTS: Forty-four of 97 eligible patients (45.4%) met criteria for rheumatologic referral. Three patients were ineligible because of elevated baseline HAQ (2) and failure to initiate AI therapy (1). No baseline characteristics were significantly associated with referral. Median time to onset of symptoms was 1.6 months (range 0.4-10 months). Clinical and laboratory evaluation of patients evaluated by rheumatology suggested that the majority developed either non-inflammatory musculoskeletal symptoms or inflammation localized to tenosynovial structures. Thirteen patients discontinued AI therapy because of musculoskeletal toxicity after a median 6.1 months (range 2.2-13 months). CONCLUSIONS: Musculoskeletal side effects were common in AI-treated patients, resulting in therapy discontinuation in more than 10% of patients. There are no identifiable pre-therapy indicators of risk, and the etiology remains elusive. PMID- 17922186 TI - Polymorphisms in three base excision repair genes and breast cancer risk in Thai women. AB - DNA repair plays an important role in tumor development. The base excision repair (BER) pathway mainly removes DNA damage caused by ionizing radiation and reactive oxidative species. Here, we examined possible associations between polymorphisms in three important BER genes (OGG1 Ser326Cys, APEX1 Asp148Glu, XRCC1 Arg194Trp, XRCC1 Arg280His, XRCC1 Arg399Gln) and breast cancer incidence in Thai women. The study population consisted of 507 breast cancer cases and 425 controls. Odds ratios (OR) were adjusted by multivariate logistic regression analysis for age, body mass index, age at menarche, family history of breast cancer, menopausal status, reproduction parameters, use of contraceptives, tobacco smoking, involuntary tobacco smoking, alcohol drinking, and education. For homozygous carriers of the Glu allele in APEX1, a significant protective effect was found when compared to Asp/Asp carriers (odds ratio (OR) = 0.60, 95% confidence interval (CI) = 0.38-0.94). Subgroup analysis based on menopausal status revealed increased breast cancer risk in postmenopausal women and OGG1 (OR = 2.05, 95% CI 1.14-3.69). Reconstructed diplotypes for XRCC1 showed that CGA/CGA carriers had an increased risk of breast cancer compared with carriers of the wild type diplotype CGG/CGG (OR = 2.56, 95% CI 1.28-5.15). When the joint effects of XRCC1, APEX1 and OGG1 polymorphisms were evaluated, individuals homozygous for two or three risk alleles were at increased risk (OR = 1.88, 95% CI 1.26-2.82). In conclusion, our data suggest that Thai women with a certain XRCC1 diplotype or homozygous for two or three variant alleles of XRCC1, OGG1, and APEX1 are likely to have an increased susceptibility to breast cancer. PMID- 17922187 TI - The CYP1B1_1358_GG genotype is associated with estrogen receptor-negative breast cancer. AB - Cytochrome P450 1B1 (CYP1B1) is a major enzyme in the initial catabolic step of estradiol (E2) metabolism and belongs to the multitude of genes regulated by the estrogen receptor alpha (ERalpha). The common non-synonymous polymorphisms CYP1B1_1358_A>G and CYP1B1_1294_C>G increase CYP1B1 enzymatic activity. Given a relationship between CYP1B1 and breast tumor E2 level as well as E2 level and breast tumor ERalpha expression it is of interest to know whether CYP1B1 polymorphisms have an impact on the ERalpha status of breast cancer. We genotyped the GENICA population-based breast cancer case-control collection (1,021 cases, 1,015 controls) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and investigated in cases the association between genotypes and tumor ERalpha status (739 ERalpha positive cases; 212 ERalpha negative cases) by logistic regression. We observed a significant association between the homozygous variant CYP1B1_1358_GG genotype and negative ERalpha status (P = 0.005; OR 2.82, 95% CI: 1.37-5.82) with a highly significant Ptrend for CYP1B1_1358_A>G and negative ERalpha status (P = 0.003). We also observed an association of CYP1B1_1358_GG and negative PR status (P = 0.015; OR 2.36, 95% CI: 1.18-4.70) and a Ptrend of 0.111 for CYP1B1_1358_A>G and negative progesterone receptor (PR) status. We conclude that the CYP1B1_1358_A>G polymorphism has an impact on ERalpha status in breast cancer in that the CYP1B1_1358_GG genotype known to encode higher CYP1B1 activity is associated with ERalpha negativity. PMID- 17922188 TI - Triple negative breast cancer: molecular profiling and prognostic impact in adjuvant anthracycline-treated patients. AB - BACKGROUND: We analysed the clinical features, distribution of basal markers, prevalence of oncogene amplification, and outcome of triple negative (TN) compared to those of non-TN cancers in a series of adjuvant-anthracycline treated breast cancer patients. METHODS: We examined the prognostic impact of the TN and BL phenotype in 245 breast cancer patients uniformly treated with adjuvant anthracycline-based chemotherapy following primary surgery, with regards to local relapse-free (LRFS), metastasis free (MFS), and breast cancer specific survival (BCSS). A comparative analysis of the clinicopathological characteristics, expression of basal markers (cytokeratins (Cks) 5/6, 14, 17, EGFR, and caveolin 1 and 2), MIB-1, p53 and topoisomerase II alpha, and prevalence of CCND1, MYC and TOP2A amplification in TN and non-TN breast tumours was performed. RESULTS: TN cancers were significantly associated with the expression of basal markers (all P < 0.0001). However 19.4% of TN tumours were negative for basal markers, whilst 7.3% of non-TN tumours expressed basal markers. TN phenotype was significantly associated with p53, MIB-1 and topoisomerase II alpha (all, P < 0.01) expression. No TN cancer harboured amplification of CCND1 or TOP2A. In univariate analysis, TN and BL phenotype were significantly associated with shorter MFS (both, P < 0.01) and BCSS (both, P < 0.005) but not LRFS. CONCLUSIONS: Despite treatment with standard dose anthracycline-based chemotherapy, the clinical outcome of TN and BL cancers remains poor. Alternative chemotherapeutic regimens and/or novel therapeutic approaches are warranted. Although a significant phenotypic overlap exists between TN and basal-like tumours, the TN phenotype is not an ideal surrogate marker for basal-like breast cancers. PMID- 17922190 TI - Prevalence and correlates of lifetime disordered gambling in Cambodian refugees residing in Long Beach, CA. AB - BACKGROUND: Research has suggested that disordered gambling is endemic to Cambodian refugees. Whereas earlier study of the prevalence and correlates of disordered gambling has relied on convenience sampling, this investigation used a subset of a sample representative of the largest Cambodian refugee community in the US. METHODS: Face-to-face interviews assessing gambling disorder were conducted with a subsample of persons (N = 127) participating in a broader study of the mental health of this community. RESULTS: 13.9% of participants met screening criteria for lifetime disordered gambling, in contrast to previous research suggesting that prevalence rates may exceed 70%. After adjusting for a range of covariates, breadth of trauma exposure and marital status emerged as significant predictors of disordered gambling. DISCUSSION: Given the myriad mental health challenges facing the Cambodian refugee community, these data indicate that scarce prevention and treatment resources may be more productively channeled toward addressing other mental health and social service needs. PMID- 17922189 TI - Effects of exercise training on inflammatory markers in patients with heart failure. AB - Cardiologists now recognize that the cardio-centric model of heart failure does not sufficiently explain the entire traits particular to chronic heart failure. Evidence accumulates, that many features of the syndrome can be explained by the known biological effects of inflammatory mediators. Indeed, when expressed in experimental models at concentrations commonly observed in heart failure, inflammatory mediators such as tumor necrosis factor-alpha, interleukin-6, and nitric oxide can produce effects that mimic features of heart failure, including (but not limited to) progressive left-ventricular dysfunction, pulmonary edema, left-ventricular remodeling, and cardiomyopathy. As we witness anti-cytokine therapies and other strategies to avoid an increase in cytokines we have been shown that acute bouts of exercise are associated with an increase in pro inflammatory cytokines and markers of oxidative stress. As a consequence we have been warned exercise may thus even further contribute to the deterioration of heart failure. However, there are several randomized trials which unanimously document that chronic--as opposed to acute bouts of--exercise does not only lead to a reduction of cytokines and oxidative stress, but that patients dramatically benefit by the increase in maximal oxygen consumption, exercise capacity, quality of life, reduction in hospitalization, morbidity, and mortality. Over the past two decades it has become evident that cytokine research has come to stay and that we will continue to see anti-cytokine treatment strategies for our patients. It is the aim of this review to shed some more light on the most commonly investigated and most relevant cytokines. PMID- 17922191 TI - Molecular mechanism of Mitomycin C-dependent caspase-8 regulation: implications for apoptosis and synergism with interferon-alpha signalling. AB - Caspase-8 is frequently mutated or silenced in several tumors including hepatocellular carcinomas (HCC) thereby potentially contributing to chemoresistance. The aim of our present study was to evaluate if chemotherapeutic drugs may mediate their effects through up-regulation of caspase-8 gene transcription. Huh7 hepatoma cells were transfected with a caspase-8 promoter construct fused to a luciferase reporter gene followed by stimulation with a subset of different chemotherapeutic drugs. Several drugs slightly induced caspase-8 promoter activity. However, strong caspase-8 promoter induction was found after Mitomycin C (MMC) treatment and this correlated with an increase in endogenous caspase-8 mRNA expression. Further molecular analysis demonstrated that MMC controls caspase-8 transcription via a c-jun/AP1 site located in the promoter in close proximity to the transcription start site. Inactivation of this c-jun/AP1 site using a dominant-negative c-jun adenovirus or site-directed mutagenesis inhibited MMC-dependent promoter induction. MMC treatment resulted in higher caspase-8 enzymatic activity and apoptosis and could be synergistically enhanced by co-stimulation with interferon-alpha (IFNalpha) via independent transcriptional mechanisms. In summary MMC controls caspase-8 expression via a c jun/AP1 element in its promoter region. MMC-induced up-regulation of caspase-8 triggers apoptosis in target cells which can be further enhanced by IFNalpha. Therefore these findings also provide a potential new therapeutic approach to treat cancer cells. PMID- 17922192 TI - Conceptualising health: insights from the capability approach. AB - This paper suggests the adoption of a 'capability approach' to key concepts in healthcare. Recent developments in theoretical approaches to concepts such as 'health' and 'disease' are discussed, and a trend identified of thinking of health as a matter of having the capability to cope with life's demands. This approach is contrasted with the WHO definition of health and Boorse's biostatistical account. We outline the 'capability approach', which has become standard in development ethics and economics, and show how existing work in those areas can profitably be adapted to healthcare. Cases are used to illustrate the value of adopting a capability approach. PMID- 17922193 TI - Representation or reason: consulting the public on the ethics of health policy. AB - Consulting the public about the ethical approaches underlying health policies can seem an appealing means of addressing concerns about limited public participation in development of health policy. However ambiguity surrounds questions of whether, or how consultation can really contribute to more defensible decisions about ethical aspects of policy. This paper clarifies the role and limits of public consultation on ethics, beginning by separating different senses of defensibility in decisions on ethics. Defensibility of ethical decisions could be understood either in the sense of legitimacy in virtue of reflecting the opinions of the public whose interests are affected, or in the sense of being able to withstand and respond to challenges presented in ethical debate. The question then is whether there are forms of consultation which have the potential to realise more defensible decisions in either of these senses. Problems of adequately accounting for the views of those affected by policy decisions casts doubt on the plausibility of using consultation as a means of determining the opinions of the public. Consultation can have a role by bringing new ideas and challenges to debate, although it is uncertain whether this will increase the defensibility of any decision on ethics. PMID- 17922194 TI - A strategy to improve priority setting in developing countries. AB - Because the demand for health services outstrips the available resources, priority setting is one of the most difficult issues faced by health policy makers, particularly those in developing countries. Priority setting in developing countries is fraught with uncertainty due to lack of credible information, weak priority setting institutions, and unclear priority setting processes. Efforts to improve priority setting in these contexts have focused on providing information and tools. In this paper we argue that priority setting is a value laden and political process, and although important, the available information and tools are not sufficient to address the priority setting challenges in developing countries. Additional complementary efforts are required. Hence, a strategy to improve priority setting in developing countries should also include: (i) capturing current priority setting practices, (ii) improving the legitimacy and capacity of institutions that set priorities, and (iii) developing fair priority setting processes. PMID- 17922195 TI - The rise of independent regulation in health care. AB - In all countries where health care access is considered a social right, regulation is both a tool of performance improvement as well as an instrument of social justice. Both social (equity in access) and economical (promoting competition) regulation are at stake due to the nature of the good itself. Different modalities of regulation do exist and usually new regulatory cycles include the creation of stronger regulatory agencies. Indeed, health care regulation is rising steadily in most developed countries as a consequence of the introduction of the New Public Management perspective to provide essential public goods. Health care is delivered by different organisations with very different cultural backgrounds--public and private (profit and non-profit)--that should be accountable for their decisions. Control by regulatory agencies is instrumental to accomplish this goal. However, there is some dispute with regards the degree of regulatory autonomy. The objective of this paper is to determine if independent regulatory agencies (IRAs) are effective in carrying out health care regulation. The authors apply Walshe's analytical framework to the Regulatory Authority of Health (Portugal) to answer the question if independent regulation works. In conclusion, the two year experience of the Regulatory Authority of Health is important not only because the primary goals of independent regulation were achieved but also because this authority is now a full partner in the health care sector. However, independent agencies need to develop strong mechanisms of accountability because good regulatory governance is the paradigm of this institutional innovation. PMID- 17922197 TI - The individual in mainstream health economics: a case of Persona Non-grata. AB - This paper is motivated by Davis' [14] theory of the individual in economics. Davis' analysis is applied to health economics, where the individual is conceived as a utility maximiser, although capable of regarding others' welfare through interdependent utility functions. Nonetheless, this provides a restrictive and flawed account, engendering a narrow and abstract conception of care grounded in Paretian value and Cartesian analytical frames. Instead, a richer account of the socially embedded individual is advocated, which employs collective intentionality analysis. This provides a sound foundation for research into an approach to health policy that promotes health as a basic human right. PMID- 17922196 TI - Genetic 'risk carriers' and lifestyle 'risk takers'. Which risks deserve our legal protection in insurance? AB - Over the past years, one of the most contentious topics in policy debates on genetics has been the use of genetic testing in insurance. In the rush to confront concerns about potential abuses of genetic information, most countries throughout Europe and the US have enacted genetics-specific legislation for insurance. Drawing on current debates on the pros and cons of a genetics-specific legislative approach, this article offers empirical insight into how such legislation works out in insurance practice. To this end, ethnographic fieldwork was done in the underwriting departments of Belgian insurance companies. Belgium was one of the first European countries introducing genetics-specific legislation in insurance. Although this approach does not allow us to speak in terms of ' the causal effects of the law', it enables us to point to some developments in insurance practice that are quite different than the law's original intentions. It will not only become clear that the Belgian genetics-specific legislation does not offer adequate solutions to the underlying issues it was intended for. We will also show that, while the legislation's focus has been on the inadmissibility of genetic discrimination, at the same time differences are made in the insurance appraisal within the group of the asymptomatic ill. In other words, by giving exclusive legal protection to the group of genetic risks, other non-genetic risk groups are unintendedly being under-protected. From a policy point of view, studying genetics-specific legislation is especially valuable because it forces us to return to first principles: Which risks deserve our legal protection in insurance? Who do we declare our solidarity with? PMID- 17922199 TI - Inequity in health care delivery in India: the problem of rural medical practitioners. AB - A considerable section of the population in India accesses the services of individual private medical practitioners (PMPs) for primary level care. In rural areas, these providers include MBBS doctors, practitioners of alternative systems of medicine, herbalists, indigenous and folk practitioners, compounders and others. This paper describes the profile, knowledge and some practices of the rural doctor in India and then discusses the reasons for lack of equity in health care access in rural areas and possible solutions to the problem. PMID- 17922198 TI - Embryonic stem cell patents and human dignity. AB - This article examines the assertion that human embryonic stem cells patents are immoral because they violate human dignity. After analyzing the concept of human dignity and its role in bioethics debates, this article argues that patents on human embryos or totipotent embryonic stem cells violate human dignity, but that patents on pluripotent or multipotent stem cells do not. Since patents on pluripotent or multipotent stem cells may still threaten human dignity by encouraging people to treat embryos as property, patent agencies should carefully monitor and control these patents to ensure that patents are not inadvertently awarded on embryos or totipotent stem cells. PMID- 17922201 TI - L.I.F.E. and D.E.A.T.H. AB - In this short, rhetorical article, I offer a thought experiment that seeks to make an analogy between 'life' and 'disease'. This article was written whilst under the influence of Nietzsche, and I hope that readers will not mistake the polemical style and the occasional nod towards humour for flippancy. This is a serious subject, and this article attempts to ask, inexplicitly, a serious question. If we do suspend our subjective value judgements about life, and strip away what might be considered the 'dogma' of value in life, what effect might this have on our feelings towards voluntary euthanasia, and what can our reaction to that thought experiment tell us? PMID- 17922200 TI - Varied and principled understandings of autonomy in English law: justifiable inconsistency or blinkered moralism? AB - Autonomy is a concept that holds much appeal to social and legal philosophers. Within a medical context, it is often argued that it should be afforded supremacy over other concepts and interests. When respect for autonomy merely requires non intervention, an adult's right to refuse treatment is held at law to be absolute. This apparently simple statement of principle does not hold true in practice. This is in part because an individual must be found to be competent to make a valid refusal of consent to medical treatment, and capacity to decide is not an absolute concept. But further to this, I argue that there are three relevant understandings of autonomy within our society, and each can demand in differing cases that different courses of action be followed. Judges, perhaps inadvertently, have been able to take advantage of the equivocal nature of the concept to come tacitly to decisions that reflect their own moral judgments of patients or decisions made in particular cases. The result is the inconsistent application of principle. I ask whether this is an unforeseen outcome or if it reflects a wilful disregard for equal treatment in favour of silent moral judgments in legal cases. Whatever the cause, I suggest that once this practice is seen to occur, acceptable justification of it in some cases is difficult to find. PMID- 17922202 TI - Heart disease prevention among Chinese immigrants. AB - There is little information about the heart disease prevention behavior of Asian immigrants. Chinese are the largest Asian sub-group in the United States (US), and 69% of Chinese Americans are foreign-born. Our objective was to describe Chinese immigrants' heart disease prevention practices. A community-based, in person survey of Chinese men and women was conducted in Seattle during 2005. Our study sample included 395 Chinese immigrants. Only 15% of the respondents consumed five or more servings of fruit/ vegetables per day, and less than one third (31%) engaged in regular physical activity. Smoking rates were significantly higher among men (21%) than women (1%). About three-quarters (74%) of the study group had received a cholesterol test in the previous five years. Recent immigrants had higher levels of fruit/ vegetable consumption and physical activity than those who had been in the US for 10 years or more. Conversely, longer duration of US residence was positively associated with recent cholesterol testing. Heart disease prevention programs should be developed, implemented, and evaluated in Chinese immigrant populations. These efforts should specifically aim to increase fruit/ vegetable consumption and regular physical activity. Future efforts to increase cholesterol testing should focus on recent immigrants. PMID- 17922203 TI - Prevalence of colorectal cancer testing and screening in a multiethnic primary care population. AB - Colorectal cancer (CRC) screening is strongly supported by evidence and widely recommended, but remains underutilized. This study reports the prevalence of CRC diagnostic testing and CRC screening in three racial/ethnic groups attending the same primary care clinic. A cross-sectional survey was conducted to elicit past history of CRC testing, including test type, indication and timing. A comparable number of African American, Hispanic and non-Hispanic white patients aged 50-80 were recruited. 560 surveys were completed: mean age was 63.4 years, 64% reported minority race/ethnicity, and 96.8% had insurance. Overall, 62.5% [95% CI: 58.5%, 66.5%] of patients were current with any type of CRC test, when diagnostic and screening procedures were included. However, 48.6% [95% CI: 44.5%, 52.7%] of the sample was current with CRC screening, when only procedures performed for screening in asymptomatic patients were included. Patients least likely to be current with testing were those of minority race/ethnicity (48.2% of Hispanics, 56.7% of African Americans and 67.5% of non Hispanic whites, p < 0.05), younger age, (57.6% of those aged 50-64, and 71.4% of those aged 65-80, p < 0.005), and those with private insurance alone (56.0% private, 67.7% public and 68.1% mixed, p < 0.05). Our findings indicate that racial/ethnic and age related disparities in CRC screening exist even in a patient population that has the same source of health care and no differences in insurance status. These results underline the need for providers to emphasize CRC screening in their practices to minority patients and those younger than 65 years of age. PMID- 17922204 TI - Knowledge, beliefs, and behaviors about hypertension control among middle-aged Korean Americans with hypertension. AB - This report describes the management and control of high blood pressure (HBP) in a sample of middle-aged (40 to 64 years) Korean Americans (KAs) with HBP. Participants in the Self-Help Intervention Program for HBP care (SHIP-HBP), a culturally tailored clinical trial for HBP management in KAs through lifestyle approaches, underwent baseline assessments to determine HBP control status and knowledge, beliefs, and practices related to HPB control. Data were stratified by sex. In all, 445 Korean patients are described. About 55% of the sample were receiving antihypertensive therapy, and less than one-third had controlled BP (< 140/90 mm Hg). Sex differences were observed in terms of clinical characteristics and anti-hypertensive medication use: women were more likely than men to have controlled BP and lower diastolic BP and to have been on HBP medication. Lower rates of smoking, drinking, and overweight or obesity were also observed more frequently in women. Most participants agreed that taking anti-hypertensive medication lowered BP. When asked what the most important behavioral factor was for controlling BP, about a quarter reported cutting down on salt, followed by exercise and medication taking. While poor HBP control remains a common problem that contributes to significant cardiovascular morbidity and mortality, particularly among ethnic minorities, our findings underscore the need to test the beneficial effects of primary and secondary prevention programs, suitably modified to meet the language, cultural, and individual needs of this high-risk population of hypertensive patients. PMID- 17922205 TI - Sexual risk behavior in men attending Mardi Gras celebrations in New Orleans, Louisiana. AB - Previous research with travelers points to higher risk behaviors during vacations. Relative to their day-to-day lives, leisure travelers have more free time to pursue sexual activities and are likely to engage in higher rates of substance use than when at home. Risk behaviors during vacation have not been thoroughly examined in men who have sex with men (MSM), a key group at risk for HIV. The present investigation examined substance use, sexual risk behaviors, and components of the Information-Motivation-Behavioral Skills (IMB) Model in MSM attending Mardi Gras celebrations in New Orleans. Almost half of the sexually active men reported having sex with a partner of unknown HIV status while in New Orleans and a similar number did not disclose their own HIV status to all of their sexual partners. Drug use and excessive alcohol use were associated with unprotected sex (ps < .05). Components of the IMB model also predicted sexual risk behavior: individuals with more accurate HIV transmission information reported fewer unprotected sex acts, and motivation to engage in sexual activity on vacation was associated with more unprotected sex (ps < .05). Findings suggest that some MSM on vacation are placing themselves at risk for HIV. Traditional HIV prevention interventions do not readily lend themselves for use with transient populations. New intervention approaches are needed to reduce sexual risk behaviors in persons traveling for leisure. PMID- 17922209 TI - High frequency transformation of the industrial erythromycin-producing bacterium Saccharopolyspora erythraea. AB - The DNA transformation in the industrial erythromycin-producing Saccharopolyspora erythraea was investigated as standard protoplast transformation methods are ineffective. Intergeneric conjugal transfer of DNA from E. coli demonstrated transformation efficiencies from 0.05 x 10(-8) to 7.2 x 10(-8) exconjugants generated per recipient. Electroporation-mediated methodologies were also established. More than 10(5) transformants were acquired per mug DNA. The proposed protocol provides an alternative route for the introduction of DNA into industrial strains. PMID- 17922206 TI - Connecting environmental health data to people and policy: integrating information and mobilizing communities for environmental public health tracking. AB - Evaluation of available data is a critical preliminary step in the assessment of local environmental health. As part of a multi-organizational initiative to improve environmental health in the Pittsburgh, Pennsylvania region, the University of Pittsburgh Center for Healthy Environments and Communities (CHEC) interviewed 70 experts in the academic, government, non-profit, and private sectors and reviewed print and electronic resources to characterize environmental and public health data available in the region. The objectives of this undertaking were: to provide a conceptual framework for categorizing data locally on environmental hazards, exposures and health endpoints, to describe and evaluate the types of environmental public health data available nationally and locally, to identify existing endeavors to gather and categorize such data, and to present case studies on the real-life relevance of the availability or lack of availability of environmental health data. The purpose and relevance of this project, the evolution of the methodology, successes and challenges met, and anticipated next steps are presented. This process description and resulting comprehensive report is available to communities, at both the state and local health department level as well as lay community members, engaged in similar endeavors, to characterize their local and regional environmental health landscape. The framework outlined serves as background for a related statewide environmental health project sponsored by the Pennsylvania Department of Health through the Pennsylvania Consortium on Interdisciplinary Environmental Policy (PCIEP) and potentially as a foundation for community-based data evaluation for the National Environmental Public Health Tracking Program. PMID- 17922210 TI - Transactivation of wound-responsive genes containing the core sequence of the auxin-responsive element by a wound-induced protein kinase-activated transcription factor in tobacco plants. AB - Mitogen-activated protein kinases (MAPKs) constitute one of the most critical signaling components in plants. A typical example is wound-induced protein kinase (WIPK), which functions during pathogen responses in tobacco plants (Nicotiana tabacum). Searching for direct down-stream components, we previously isolated a novel transcription factor, which was activated upon phosphorylation by WIPK and designated as N. tabacum WIPK-interacting factor (NtWIF). Overexpression of NtWIF in tobacco plants enhanced the hypersensitive response (HR) upon tobacco mosaic virus infection and cryptogein treatment, while its silencing by RNAi suppressed such HR. NtWIF contains a specific motif similar to the B3 DNA binding domain, which recognizes the core TGTCTC motif called the auxin-responsive element (ARE). Using synthetic ARE sequences, NtWIF was also shown to recognize the ARE motifs and to transactivate the Luciferase (Luc)-reporter gene driven by such AREs in tobacco BY2 cultured cells. Subsequent microarray screening of NtWIF overexpressing tobacco identified 49 stress-responsive genes, and in silico analyses of available promoter regions of these genes revealed beta-1,3 glucanase, ACS2, P-450, and WIPK itself to contain the ARE core motif consisted of either TGTCTC or TGTCCT. Gel shift assay showed NtWIF to efficiently bind to both sequences. Assays with 1.5-kb PR-Q and 1.2 kb WIPK promoter regions, each fused to the Luc-reporter gene, indicated NtWIF to exhibit a clear transactivation activity, which was increased up to 3-fold upon phosphorylation by WIPK. These results revealed that NtWIF directly regulates multiple stress responsive genes containing the ARE motif in their promoters, thereby partly filling up the last step of the MAPK cascade. PMID- 17922211 TI - Degradation of pyridine and 4-methylpyridine by Gordonia terrea IIPN1. AB - Gordonia terrea IIPN1 was isolated and characterized from soils collected at petroleum drilling sites. The strain was able to catabolize pyridine and 4 methylpyridine as sole carbon and nitrogen source. The strain failed to catabolize other pyridine derivatives. Growing cells completely degraded 30 mM of pyridine in 120 h with growth yield of 0.29 g g(-1). Resting Cells grown on 5 mM pyridine degraded 4-methylpyridine without a lag time and vice versa. Supplementary carbon and nitrogen source did not significantly change the specific growth rate and degradation rate by the resting cells. PMID- 17922212 TI - Statistical evaluation of hydrobiological parameters of Narmada River water at Hoshangabad City, India. AB - Narmada is considered to be the lifeline of the state of Madhya Pradesh in Central India. The Narmada water is used for bathing, drinking, irrigation and industrial purposes. The city sewage and industrial effluent from Security paper mill at Hoshangabad drains in the Narmada River and pollutes the water quality. Urban sewage enters into Narmada through main nallas. River water quality at Hoshangabad has become a matter of concern due to continuous changing environment and increasing social and industrial activity that influence the water quality directly or indirectly. The present investigation is undertaken to study the effect of domestic sewage and effluent from Security paper Mill on the water quality and ecology of river Narmada at Hoshangabad. The study is carried on at four sites along with the bank of river Narmada. Water samples from four stations were collected, out of which three main sewage mixing points of the city and one fresh water site are taken into account. The samples collected were analyzed, as per standard methods parameters such as Temperature, pH, were measured in-situ. The statistical evaluations were also made. The result showed increase in BOD, Nitrates, Phosphates and Total Coliforms, No. of phytoplanktons. The results revealed that most of the water samples were below or out of limited; according to the WHO, BIS standards. PMID- 17922213 TI - Electrostatic effect of surfactant molecules on bacteriochlorophyll a and carotenoid binding sites in the LH1 complex isolated from Rhodospirillum rubrum S1 probed by Stark spectroscopy. AB - The LH1 complexes were isolated from the purple photosynthetic bacterium Rhodospirillum rubrum strain S1. They were initially solubilized using LDAO and then purified in the presence of Triton X-100. The purified complexes were then either used directly or following an exchange into LDAO. Stark spectroscopy was applied to probe the electrostatic field around the bacteriochlorophyll a (BChl a) and carotenoid binding sites in the LH1 complexes surrounded by these two different surfactant molecules. Polarizabilty change (deltaalpha)) and dipole moment change (deltamicrom) upon photoexcitation were determined for the BChl a Q(y) band. Both of these parameters show smaller values in the presence of LDAO than in Triton X-100. This indicates that polar detergent molecules, like LDAO, affect the electrostatic environment around BChl a, and modify the nonlinear optical parameters (deltaalpha and deltamicrom values). The electrostatic field around the BChl a binding site, which is generated by the presence of LDAO, was determined to be |E ( L )| = approximately 3.9 x 10(5) [V/cm]. Interestingly, this kind of electrostatic effect was not observed for the carotenoid-binding site. The present study demonstrates a unique electrostatic interaction between the polar detergent molecules surrounding the LH1 complex and the Q(y) absorption band of BChl a that is bound to the LH1 complex. PMID- 17922214 TI - Phycobilisome-reaction centre interaction in cyanobacteria. AB - The phycobilisome is a remarkable light-harvesting antenna that combines high efficiency with functional flexibility and the ability to capture light across a broad spectral range. A combination of biochemical, structural and spectroscopic studies has given an excellent picture of the structure and function of isolated phycobilisomes. However, we still know remarkably little about the interaction of the phycobilisome with the thylakoid membrane and the reaction centres. This article will discuss the various current ideas about this question and explain the things we need to know more about. As a working model, I propose that the phycobilisome is attached to the membrane by multiple weak charge-charge interactions with lipid head-groups and/or proteins, and that the core-membrane linker polypeptide ApcE provides a flexible surface allowing interaction with multiple membrane components. PMID- 17922215 TI - Ribulose-1,5-bisphosphate carboxylase/oxygenase from thermophilic cyanobacterium Thermosynechococcus elongatus. AB - Ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) can be divided into two branches: the "red-like type" of marine algae and the "green-like type" of cyanobacteria, green algae, and higher plants. We found that the "green-like type" rubisco from the thermophilic cyanobacterium Thermosynechococcus elongatus has an almost 2-fold higher specificity factor compared with rubiscos of mesophilic cyanobacteria, reaching the values of higher plants, and simultaneously revealing an improvement in enzyme thermostability. The difference in the activation energies at the transition stages between the oxygenase and carboxylase reactions for Thermosynechococcus elongatus rubisco is very close to that of Galdieria partita and significantly higher than that of spinach. This is the first characterization of a "green-like type" rubisco from thermophilic organism. PMID- 17922216 TI - Complete recovery from acute encephalopathy of late-onset ornithine transcarbamylase deficiency in a 3-year-old boy. AB - Ornithine transcarbamylase deficiency is the commonest urea cycle disorder which is transmitted in X-linked inheritance. It is mainly characterized in males by acute encephalopathy and hyperammonaemia with fatal outcomes in both classical neonatal and late-onset types. We report a 3-year-old healthy Hong Kong Chinese boy who presented with acute encephalopathy and coma after three days of gastroenteritis. He had no focal neurological deficit and brain CT imaging was normal. His plasma ammonia (54 micromol/L) and glutamine (747 micromol/L) concentrations were normal. The only biochemical abnormalities detected were marked orotic aciduria (700 micromol/mmol creatinine) and elevated urinary uracil. He regained consciousness spontaneously after three days under intensive care with parenteral fluid therapy. He recovered completely without any neurological deficits. Five months after discharge, urinary uracil concentration remained elevated despite normalized orotic acid concentration. Finally, ornithine transcarbamylase deficiency was diagnosed by DNA analysis. A missense mutation of arginine-to-glutamine substitution on amino acid 277 (p.R277Q) was revealed to be a late-onset mutant. Our case strengthens the argument that in any child with coma or acute encephalopathy of undetermined cause, genetic analysis of the OTC gene and the measurement of urinary uracil concentration remain the most reliable indicators of late-onset OTCD during acute and even quiescent phases. Existing neonatal screening programmes for inheritable metabolic disorders fail to detect late-onset variants. Therefore, a high clinical suspicion is a key to correct and timely diagnosis, especially in those patients with atypical presentations. PMID- 17922217 TI - Description of the mutations in 15 subjects with variant forms of maple syrup urine disease. AB - BACKGROUND: In maple syrup urine disease (MSUD), disease-causing mutations can affect the BCKDHA, BCKDHB or DBT genes encoding for the E1 alpha, E1 beta and E2 subunits of the multienzyme branched-chain 2-keto acid dehydrogenase (BCKD) complex. AIM: The aim of this study was to screen DNA samples of 15 subjects with distinct well-characterized variant MSUD phenotypes for mutations in the three genes in order to demonstrate a potential correlation between specific nucleotide changes and particular variant phenotypes. METHODS: The exonic coding sequences of all three genes were studied using genomic DNA and cellular RNA derived from peripheral blood leukocytes. RESULTS: In 37% of the cases (total 30 alleles), disease-causing mutations were located in the BCKDHA, in 46% in the BCKDHB, and in 13% in the DBT gene. Novel mutations occurring homozygously were p.Ala328Thr in the BCKDHA gene and p.Gly249_Lys257del in the DBT gene. Both are associated with a mild MSUD variant. The same holds true for the novel mutations p.Pro200Ala in BCKDHB and p.Phe307Ser in DBT which were identified in heterozygous fashion. Among the known mutant alleles, p.Gly278Ser in the BCKDHB gene was relatively frequent and also associated with a mild MSUD variant. CONCLUSION: The results of this study indicate that genotyping may be predictive of clinical severity of variant MSUD phenotypes and might be of prognostic value particularly in subjects with variant MSUD identified in newborn screening in whom early treatment fortunately slows the natural course of the disease. PMID- 17922219 TI - Mycetomas in Iran: a review article. AB - Mycetomas are the subcutaneous and relatively rare chronic pustular infections. The etiologic agents of mycetomas are a group of saprophytic fungi and actinomycetes living in soil. We retrospectively discussed the overall prevalence of mycetomas and the prevalence of infective agents in Iran between 1972 and 2005. Seventy-six cases of mycetomas have been reported from various geographical locations in Iran during 33 years. Analysis of the records revealed that 84.5% were actinomycetoma and only 15.5% were eumycetoma. Disease mainly has been seen in foot, and the male to female ratio was 2:1. Mycetomas were abundant among farmers in rural areas of Iran. The commonest agents of mycetomas were Nocardia asteroids, Actinomadura madura (actinomycetoma) and Allesheria boydii (eumycetoma). The peak age of onset was between 31 and 51 years. PMID- 17922218 TI - Effect of pre-incubation temperature on susceptibility of Galleria mellonella larvae to infection by Candida albicans. AB - The use of insects for evaluating the virulence of microbial pathogens and for determining the efficacy of antimicrobial drugs is increasing. When larvae of the greater wax moth Galleria mellonella were incubated at 4 or 37 degrees C for 24 h. prior to infection, they manifested increased resistance to infection by the yeast Candida albicans compared to larvae that had been pre-incubated for 24 h at 30 degrees C. Incubation at 4 or 37 degrees C led to an increase in haemocyte density and the expression of genes coding for gallerimycin, transferrin, an inducible metalloproteinase inhibitor (IMPI) and galiomicin. Peak expression of these genes was recorded at approximately 24 h after the commencement of the 4 or 37 degrees C incubation. These results indicate that exposure of larvae to mild thermal shock conditions induces a protective cellular and humoral immune response mediated by increased numbers of haemocytes and elevated expression of antimicrobial peptides. PMID- 17922221 TI - A randomized validation study comparing embedded versus extracted FACT Head and Neck Symptom Index scores. AB - OBJECTIVE: To evaluate the impact of administration context (embedded versus stand-alone) on the reliability and validity of the FACT Head and Neck Symptom Index (FHNSI). METHODS: Ninety-eight patients with head and neck cancer were randomized to one of two assessment conditions to evaluate the FHNSI's context (items administered embedded within the FACT-H&N or as stand-alone scale) and order of administration in the battery. RESULTS: Planned comparisons on the item and scale levels revealed no systematic order or context differences. The embedded and stand-alone versions of the FHNSI showed high internal consistency (Cronbach's alpha 0.79-0.87). Correlations were high between the FHNSI versions and the physical and functional well-being scales of the FACT-H&N (0.70-0.84) and measures of pain intensity (-0.73, -0.74) and depression (-0.71, -0.74); moderate to large with the Performance Status Scale for Head and Neck subscales (PSS-HN; 0.46-0.71); and low with an anxiety measure (0.30, 0.34). Both FHNSI versions differentiated patients grouped by performance status (p < .0001, p < .0001) and global rating of change (p < .0001, p < 0.01). The FHNSI's minimally important difference range was 3-4 points. CONCLUSION: The FHNSI is a reliable and valid symptom index, which can be administered alone or scored using items embedded within the FACT-H&N. PMID- 17922220 TI - Identification and characterization of two chrysanthemum (Dendronthema x moriforlium) DREB genes, belonging to the AP2/EREBP family. AB - Two DREB-like genes, designated as DmDREBa and DmDREBb, were isolated from chrysanthemum by RACE approach. The DmDREBa and DmDREBb genes encoded proteins of 191 and 185 amino acid residues with the predicted molecular mass of 21.66 and 20.99 kDa, respectively. Both DmDREBs proteins comprised a typical EREBP/AP2 domain, a DREB1-type nuclear localization signal (NLS) before the EREBP/AP2 domain and a DSAWR sequence after the domain. Phylogenetic analysis suggested that both DmDREBs were attributable to the DREB1 subgroup. Quantitative real-time PCR analysis revealed that both genes were accumulated more in leaves and stems than in roots and flowers. Moreover, DmDREBb reacted earlier and accumulated with higher levels than DmDREBa under cold treatment. Expression of DmDREBa and DmDREBb declined dramatically within 0.5 h of exposure to 100 microM ABA. Besides, DmDREBb expression was variable and recovered to pre-ABA levels at 2, 6 and 12 h, while DmDREBa expression remained low during the 24 h exposure period. Furthermore, both genes expression was totally inhibited at 40 degrees C. In addition, these two DmDREB proteins were also found to have transcriptional activity and had the DRE-binding capacity as shown by yeast one-hybrid system and were localized to the nuclei of cells. To our knowledge, this is the first report on isolation of DREB-like genes in chrysanthemum. This study showed that DmDREBs may play a regulatory role in abiotic stress responses and provided evidence of differences in the timing and level of expression between two homologous genes in plants under abiotic stress conditions. PMID- 17922222 TI - Spontaneous reports of most distressing concerns in patients with inoperable lung cancer: at present, in retrospect and in comparison with EORTC-QLQ-C30+LC13. AB - Patients with lung cancer experience considerable distress. Therefore, accurate methods for assessing distress and quality of life over time may play a key role for managing and evaluating palliative care. Alternatives to commonly used standardized questionnaires are individual measures. This study prospectively and retrospectively explored the concerns that 46 patients with inoperable lung cancer spontaneously reported as causing most distress close to diagnosis and 6 months later. Changes in content individually generated through a structured inductive freelisting were compared with EORTC-QLQ-C30+LC13 ratings. The results showed that patients perceived a wide variety of concerns as most distressing and that their concerns changed over time. Between 56 and 62% of these concerns were assessed by items included in the EORTC-QLQ-C30+LC13 questionnaires. Furthermore, patients' reports of most distress from fatigue, pain and dyspnea were not always reflected in intensity ratings of comparable EORTC-QLQ-C30+LC13 items. These results indicate that items included in standardized measures are not always adequate to assess patients' concerns, priorities and changes over time. In addition to standardized questionnaires, individualized measures may be useful in the clinical palliative setting for providing detailed information about the individual's problems and prioritizations. PMID- 17922224 TI - A novel synthetic oleanolic acid derivative (CPU-II2) attenuates liver fibrosis in mice through regulating the function of hepatic stellate cells. AB - Regulation on the function of the hepatic stellate cells (HSCs) is one of the proposed therapeutic approaches to liver fibrosis. In the present study, we examined the in vitro and in vivo effects of CPU-II2, a novel synthetic oleanolic acid (OLA) derivative with nitrate, on hepatic fibrosis. This compound alleviated CCl4-induced hepatic fibrosis in mice with a decrease in hepatic hydroxyproline (Hyp) content and histological changes. CPU-II2 also attenuated the mRNA expression of alpha-smooth muscle actin (alpha-SMA) and tissue inhibitor of metalloproteinase type 1 (TIMP-1) induced by CCl4 in mice and reduced both mRNA and protein levels of alpha-SMA in HSC-T6 cells. Interestingly, CPU-II2 did not affect the survival of HSC-T6 cells but decreased the expression of procollagen alpha1 (I) in HSC-T6 cells through down-regulating the phosphorylation of p38 MAPK. CONCLUSION: CPU-II2 attenuates the development of liver fibrosis rather by regulating the function of HSCs through p38 MAPK pathway than by damaging the stellate cells. PMID- 17922223 TI - Is MSH2 a breast cancer susceptibility gene? AB - Mutations in the DNA mismatch repair gene MSH2 lead to increased replication error and microsatellite instability and account for a substantial proportion of hereditary non-polyposis colorectal cancer (Lynch syndrome). A recent international collaborative genome-wide linkage scan (GWS) for breast cancer susceptibility loci found some evidence for there being a breast cancer susceptibility gene in a genomic region on chromosome 2p close to MSH2. We sought to investigate the possibility that mutations in MSH2 might explain the multiple cases of breast cancer in some families that were included in the international GWS. DNA samples from the affected probands of 59 multiple-case breast cancer families, many of whom gave LOD scores >0.5 in the MSH2 region, were screened for large genomic alterations in MSH2 via the Multiplex Ligation-dependent Probe Amplification (MLPA) assay and for coding region mutations via exonic sequencing. Several of the families also contained cases of colorectal cancer in addition to breast cancer and had been included in the GWS that had identified a positive LOD score on chromosome 2p. Using MLPA, c.1236C > T was identified in one proband but this variant was not predicted to create an alternate acceptor/donor site within exon 7 MSH2 using in silico analyses. A c.1734T > C was identified in a second proband via exonic sequencing but testing of the variant in other family members did not support segregation of this variant with disease. Extensive screening of 59 multiple-case breast cancer families did not identify any coding region mutations or larger genomic alterations in MSH2 that might implicate MSH2 as a breast cancer susceptibility gene. PMID- 17922225 TI - Androgen receptor gene polymorphism may affect the risk of urothelial carcinoma. AB - The study sought to explore if androgen receptor gene (AR) polymorphisms are associated with the risk of urothelial carcinoma (UC) which is male-predominant. AR CAG and GGN repeat lengths were analyzed in 277 UC cases and 280 age and sex matched controls by direct sequencing of leukocyte DNA. Smoking habits were obtained using a structured questionnaire interview. Relative risks were compared between groups categorized by all possible cutoffs of AR CAG and GGN repeat lengths. Men and women who had 23 and 44 (cumulative) CAG repeats had a significantly greater risk of UC, respectively (OR 2.09, 95% CI: 1.05-4.17, p = 0.036 and OR 4.95, 95% CI: 1.56-15.73, p = 0.007). Amongst males who were medium dose cigarette smokers, those who had 23 CAG and shorter GGN (<22) repeats, had an elevated risk than those with longer CAG and GGN (OR 4.32 and 4.57, p = 0.034 and 0.042, respectively). However, neither CAG nor GGN affected the UC risk in non-smokers or heavy smokers (> or =25 packs per day-years). AR CAG polymorphism may affect the risk of UC in both genders. In addition, AR polymorphisms may influence carcinogenic effect of medium-dose of cigarette smoking in men. PMID- 17922226 TI - PCR amplification of microsatellites from single cells of Karenia brevis preserved in Lugol's iodine solution. AB - A simple and effective protocol is described for multiplex polymerase chain reaction (PCR) amplification of single cells of Karenia brevis. The protocol requires minimum processing, avoids additions that might dilute target DNA template, and can be used on cells preserved in Lugol's iodine preservative. Destaining of Lugol's-preserved cells with sodium thiosulfate allowed successful amplification of single-copy, nuclear-encoded microsatellites in single cells of K. brevis that have been preserved for up to 6 years. PMID- 17922227 TI - Reducing local recurrence after breast-conserving surgery for breast cancer. PMID- 17922228 TI - Podocalyxin enhances the adherence of cells to platelets. AB - Podocalyxin (PODXL) is a mucin protein of the CD34 family expressed in kidney glomerular podocytes, vascular endothelium, progenitor bone marrow and tumor cells. It is assumed that PODXL plays an anti-adherent role in kidney podocytes. CHO cells stably expressing human PODXL (CHO-PODXL) or human tumor cells (Tera-1) inherently expressing PODXL showed increased adherence to platelets. The adherence of cells was inhibited (70%) by blockers of platelet P-selectin, prevented by the soluble ectodomain of human PODXL (PODXL-Delta) or by the arginine-glycine-aspartate (RGDS) peptide and partially impeded by inhibition of integrin alphaVbeta3/alphaVbeta5, suggesting a coordinated action of P-selectin and integrins. Colocalization of platelet P-selectin and PODXL expressed on CHO cells was demonstrated by confocal immunofluorescence. No adherence to platelets was observed when PODXL was expressed in glycomutant CHO cells deficient in sialic acid. PMID- 17922229 TI - Arachidonic acid signaling is involved in the mechanism of imidazoline-induced KATP channel-independent stimulation of insulin secretion. AB - The mechanism by which the novel, pure glucose-dependent insulinotropic, imidazoline derivative BL11282 promotes insulin secretion in pancreatic islets has been investigated. The roles of KATP channels, alpha2-adrenoreceptors, the I1 receptor-phosphatidylcholine-specific phospholipase (PC-PLC) pathway and arachidonic acid signaling in BL11282 potentiation of insulin secretion in pancreatic islets were studied. Using SUR1(-/-) deficient mice, the previous notion that the insulinotropic activity of BL11282 is not related to its interaction with KATP channels was confirmed. Insulinotropic activity of BL11282 was not related to its effect on alpha2-adrenoreceptors, I1-imidazoline receptors or PC-PLC. BL11282 significantly increased [3H]arachidonic acid production. This effect was abolished in the presence of the iPLA2 inhibitor, bromoenol lactone. The data suggest that potentiation of glucose-induced insulin release by BL11282, which is independent of concomitant changes in cytoplasmic free Ca2+ concentration, involves release of arachidonic acid by iPLA2 and its metabolism to epoxyeicosatrienoic acids through the cytochrome P-450 pathway. PMID- 17922230 TI - Nutrigenomics in the whole-genome scanning era: Crohn's disease as example. AB - Nutrigenomics has the potential to tailor diets to optimize health, based on knowledge of key genetic polymorphisms. Identification of candidate genes is often based on a priori knowledge of disease processes. However, genome-wide association methods are not only validating previously identified genes and polymorphisms, but also revealing new gene-disease associations not anticipated from prior knowledge. In Crohn's disease (CD), such studies not only confirm the importance of caspase-activated recruitment domain 15 and major histocompatibility complex II molecules, but also reveal strong associations with the proinflammatory cytokine interleukin-23 receptor and autophagy-related 16 like gene. Genes identified to date in CD can be linked into two interrelated pathways: receptor-mediated cytokine induction or autophagocytosis. New genomic technologies need to be matched with innovative methodologies to characterize the likely impact of foods and to take the field to another dimension of value for human diet development and optimized health. PMID- 17922231 TI - Polymorphisms in manganese superoxide dismutase, myeloperoxidase and glutathione S-transferase and survival after treatment for metastatic breast cancer. AB - Treatments for metastatic breast cancer (MBC) are primarily palliative with variable efficacy and outcomes may be influenced by individual differences in drug metabolism. In this study, we examined the association of single nucleotide polymorphisms (SNPs) in genes involved in drug metabolism with progression free survival (PFS) and breast cancer specific survival (BCSS) in 95 patients with MBC that received high dose chemotherapy (HDC) with autologous stem cell transplantation (ASCT). SNPs in the SOD2 (SOD2-01, Val16Ala), MPO (MPO-02, -463 promoter variant) and GSTP1 [GSTP1-01 (Ile105Val), GSTP1-02 (Ala114Val)] genes were examined in DNA isolated from cryopreserved blood products using genotyping assays. Survival was analysed using Cox proportional hazard models and Kaplan Meier estimates. Patients with the SOD2-01 (TT) genotype had increased risk of disease progression [hazard ratio (HR): 2.52; 95% confidence interval (CI), 1.31 4.85] and breast cancer specific death (HR: 1.92; 95% CI: 1.03-3.57). Risks were increased for patients with both SOD2-01 (TT) and GSTP1-01 (GG or AG) genotypes (HR for disease progression: 2.57, 95% CI: 1.32-5.00 and HR for breast cancer specific death: 2.27; 95% CI: 1.18-4.34). In multivariable analysis, the combined genotype group of SOD2-01 and GSTP1-01 was an independent predictor of PFS and BCSS. HRs progressively increased with increasing number of genotypes associated with worse survival, with p(trend) of 0.005 and 0.006 for PFS and BCSS, respectively. These results suggest that SNPs in genes involved in drug metabolism may influence survival outcome for patients with MBC receiving HDC and ASCT. PMID- 17922232 TI - Parent-child acculturation discrepancies as a risk factor for substance use among Hispanic adolescents in Southern California. AB - Theories of acculturation predict that discrepancies in cultural orientation between adolescents and their parents will increase the adolescents' risk for behavior problems such as substance use. This study evaluated this hypothesis in a sample of 1772 Hispanic 9th grade students in Southern California. Parent-child discrepancy in U.S. orientation (defined as the difference between the child's U.S. orientation and the child's perception of the parents' U.S. orientation) was a risk factor for past-month smoking, lifetime and past-month alcohol use, and lifetime and past-month marijuana use. Parent-child discrepancy in Hispanic orientation (defined as the difference between the child's Hispanic orientation and the child's perception of the parents' Hispanic orientation) was a risk factor for lifetime and past-month alcohol and marijuana use. The adolescents' own Hispanic orientation was protective against lifetime and past-month smoking and marijuana use, but not alcohol use. In an analysis of mediation, U.S. acculturation discrepancy was associated with lower levels of family cohesion, which in turn was associated with higher levels of substance use. Results suggest that family-based interventions for acculturating and bicultural Hispanic families may be useful in decreasing the likelihood of substance use among Hispanic adolescents. PMID- 17922233 TI - Toxic metals and organochlorine pesticides residue in single herbal drugs used in important ayurvedic formulation - 'Dashmoola'. AB - Herbal formulations are getting popularity throughout the world and commercialized extensively for various medicinal properties. WHO has emphasized the need for quality assurance of herbal products, including testing of heavy metals and pesticides residues. 'Dashmoola', a popular herbal formulation, with immunomodulator and febrifugal properties, consists of ten single root drugs. In view of WHO guidelines, single herbal drugs used in 'Dashmoola', were collected from different places of India for testing heavy metals and persistent pesticides residue. Although use of roots in 'Dashmoola' is prescribed in original ayurvedic literature but now many pharmacies use stem in place of roots. Therefore, in the present study both roots and stems were selected for estimation of six heavy metals namely arsenic (As), mercury (Hg), lead (Pb), cadmium (Cd), chromium (Cr) and nickel (Ni). Apart from these, the organochlorine pesticides residue viz. different metabolites of DDT, DDE, isomers of HCH and alpha-endosulfan were checked in total 40 samples of single crude drugs. Heavy metals except Hg, were present in most of the samples. In few samples Pb and Cd concentration were beyond the WHO permissible limits. Although alpha-HCH and gamma-HCH were present in almost all the samples, but other pesticides were not detected in these samples. DDT and DDE were found only in two samples. PMID- 17922235 TI - Strong larvicidal activity of three species of Spilanthes (Akarkara) against malaria (Anopheles stephensi Liston, Anopheles culicifacies, species C) and filaria vector (Culex quinquefasciatus Say). AB - A system for biocontrol of malaria and filarial mosquito vectors has been developed using herbal extracts of three Spilanthes species, S. acmella L.var oleraceae Clarke, S. calva L. and S. paniculata Wall ex DC. Cent percent mortalities was achieved against the late third/early fourth instar larvae of A. stephensi Liston, A. culicifacies species C and C. quinquefasciatus Say using crude hexane extract obtained from flower heads of Spilanthes spp. Of the three plant species, S. acmella extract proved to be the most effective in inducing complete lethality at minimum doses, the respective LC50 and LC90 values being 4.57 and 7.83 (A. stephensi), 0.87 and 1.92 (A. culicifacies) and 3.11 and 8.89 ppm (C. quinquefasciatus). This was followed by S. calva and S. paniculata extracts, respectively. This is the first report of achieving cent percent lethality against these mosquito larvae using minimal doses of plant extracts from this or any other plant species. PMID- 17922234 TI - Mescaline effects on rat behavior and its time profile in serum and brain tissue after a single subcutaneous dose. AB - RATIONALE: Mescaline is a nonselective serotonin receptor agonist. It has relatively delayed onset of action and prolonged duration. Mescaline attenuates various behavioral parameters in rats; however, no information is available about its pharmacokinetics in rats and its relation to the behavioral changes produced by the drug. OBJECTIVES: The present study evaluates the spontaneous locomotor activity and sensorimotor gating in relation to mescaline concentrations in the serum and the brain of rats MATERIALS AND METHODS: Behavioral changes induced by mescaline [10, 20, and 100 mg/kg subcutaneously (s.c.)] were evaluated in an open field test and testing of the prepulse inhibition of acoustic startle reaction (PPI) 15 and 60 min after drug administration. The time disposition of mescaline 20 mg/kg s.c. in rat serum and brain homogenates was analyzed by gas chromatography-mass spectrometry. RESULTS: Mescaline produced significant inhibitory effects on locomotion in low doses and a biphasic effect with the highest dose. In the PPI test, only when tested 60 min after drug administration, all doses of mescaline disrupted PPI. Besides the experimental protocol, we have observed that approximately 50% of animals receiving 100 mg/kg died within 12 h post-injection. The serum levels of mescaline rapidly increased within 30 min and subsequently quickly decreased; however, the brain concentrations reached a maximum 1 h after administration and remained high for an additional 60 min. CONCLUSIONS: Mescaline had a delayed onset of the main behavioral changes in rats compared to other hallucinogens. Behavioral changes correlated with the pharmacokinetics of the drug. PMID- 17922237 TI - Variants of personality maladaptation in patients with multiple sclerosis. AB - A total of 34 patients with multiple sclerosis (MS) aged 14-52 years, with disease onset at age 24.3 +/- 7.5 years, were studied. Disease severity on the Kurtzke scale (EDSS) was 3.6 +/- 1.7 points. Seven patients were in exacerbation of MS and 27 were in remission. Personality characteristics in terms of MS associated maladaptation were studied using a modified MMPI (the SMPT method). Neurotic, psychotic, and mixed types of maladaptation were identified. Measures of brain metabolic activity were simultaneously determined (in terms of the rate of glucose utilization) by positron emission tomography (PET). Data were obtained on the relationship between the activity of metabolic processes in the frontal, temporal-parietal, and limbic areas of the cerebral cortex and different variants of personality maladaptation and, respectively, with different personality profiles in the SMPT. PMID- 17922236 TI - Genomic expression patterns in cell separation mutants of Schizosaccharomyces pombe defective in the genes sep10 ( + ) and sep15 ( + ) coding for the Mediator subunits Med31 and Med8. AB - Cell division is controlled by a complex network involving regulated transcription of genes and postranslational modification of proteins. The aim of this study is to demonstrate that the Mediator complex, a general regulator of transcription, is involved in the regulation of the second phase (cell separation) of cell division of the fission yeast Schizosaccharomyces pombe. In previous studies we have found that the fission yeast cell separation genes sep10 ( + ) and sep15 ( + ) code for proteins (Med31 and Med8) associated with the Mediator complex. Here, we show by genome-wide gene expression profiling of mutants defective in these genes that both Med8 and Med31 control large, partially overlapping sets of genes scattered over the entire genome and involved in diverse biological functions. Six cell separation genes controlled by the transcription factors Sep1 and Ace2 are among the target genes. Since neither sep1 ( + ) nor ace2 ( + ) is affected in the mutant cells, we propose that the Med8 and Med31 proteins act as coactivators of the Sep1-Ace2-dependent cell separation genes. The results also indicate that the subunits of Mediator may contribute to the coordination of cellular processes by fine-tuning of the expression of larger sets of genes. PMID- 17922238 TI - Clinical-psychological characteristics of children with dysgenesis of the cerebellar vermis. AB - This report addresses behavioral abnormalities in children with cerebellar anomalies demonstrated on MRI scans. Published data are presented showing an interaction between cerebellar pathology and early childhood autism. The cerebellum is involved not only in movement coordination, but also in social adaptation and verbal communication. The genes expressed in the cerebellum during childhood are identical to those expressed in the hippocampus. We have observed 20 children with MRI-identified agenesis of the cerebellar vermis and behavioral abnormalities; children were aged 3-15 (mean 7.05) years and there were 12 males and eight females. A variety of autistic characteristics were identified in these children. PMID- 17922239 TI - Glutamate receptor autoantibody concentrations in children with chronic post traumatic headache. AB - We report here studies on the levels of autoantibodies (aAb) to AMPA glutamate receptors (GluR1 subunit) and NMDA glutamate receptors (NR2A subunit) in serum from 60 children aged 7-16 years with chronic posttraumatic headache (CPTHA) following mild craniocerebral trauma (CCT). The first group consisted of 48 children who had sustained cerebral concussion (CC), of which 34 had single episode CC (subgroup 1a) and 14 had repeated CC (subgroup 1). The second group included 12 children with mild cerebral contusions (MCC). Serum glutamate receptor aAb levels were measured six months and one year after trauma. Increased aAb levels were expressed as percentages and were regarded as significant when increases were to 120% of the level seen in healthy children of the same age. The highest levels of aAb to NMDA receptors were seen in children with MCC (165 +/- 34%) and single CC (145 +/- 12.6%). Children with repeated CC had NMDA receptor aAb at normal levels (108 +/- 12.4%). Increases in NMDA receptor aAb were seen during the first year after trauma. Increases in AMPA receptor aAb were seen in children with repeated CC and MCC (150 +/- 16.8% and 167 +/- 31.3%). EEG studies showed that 18% of these children had nonspecific paroxysmal changes and 6% showed epileptiform activity. These results provide evidence that children with post-traumatic headache demonstrated hyperstimulation of glutamate receptors and overdevelopment of the autoimmune process. Increases in serum levels of aAb to NMDA glutamate receptors reflected hypoxic-ischemic brain lesions in children with CPTHA and dictate the need for these children to receive metabolic therapy. PMID- 17922240 TI - Kindling in the early postnatal period: Effects on the dynamics of age-related changes in electrophysiological characteristics of hippocampal neurons. AB - The effects of chronic administration of pentylenetetrazole (PTZ) during early ontogenesis (from postnatal day 14) on the dynamics of age-related changes in electrophysiological characteristics of rat hippocampal slices were studied. Unlike the situation in adult animals, convulsive activity did not develop in rat pups in response to repeated injections. Comparison of the amplitude characteristics of total monosynaptic responses of neurons in hippocampal field CA1 to application of single and paired (separated by 70 msec) stimulation of Schaffer collaterals during the period of maximally intense hippocampal synaptogenesis (at weeks 2-3 of postnatal development) revealed no significant differences between the control group (administration of isotonic saline) and the group given PTZ. The level of suppression of facilitation in paired-pulse stimulation with a short interstimulus interval (15 msec) was significantly less in hippocampal slices from rat pups from the PTZ group. However, as compared with the passive control, the direction of rearrangements in the two experimental group was essentially the same. Nonetheless, regular administration of PTZ during the period of maximally intense hippocampal maturation affected the development of its characteristics. This was not only apparent as a deficiency of inhibitory processes. Increases in the intensity of test stimuli applied to hippocampal slices from PTZ-treated rat pups at 27-48 days of age led to relatively lower response amplitudes as compared with those seen in hippocampal slices from control (given isotonic saline) rats of the same age. The level of facilitation in paired-pulse stimulation with an interstimulus interval of 70 msec showed no difference, decreasing to similar extents in both groups as compared with the passive control group. In addition, hippocampal slices from the PTZ group showed significant decreases in the magnitude of long-term potentiation. Changes occurring in the hippocampus after regular administration of PTZ did not correlate with the development of convulsive activity. The only significant relationship involving the intensity of convulsions was with the increase (compared with the normal for age) in the amplitudes of responses to minimal intensity test stimuli. PMID- 17922241 TI - Recording of spontaneous oscillations in the procerebrum of the terrestrial snail Helix during free behavior. AB - The procerebrum is the central component of the olfactory analyzer in terrestrial snails and slugs; spontaneous rhythmic oscillations have been recorded in this structure, but their role in the mechanisms of odor perception and recognition has yet to be clarified. The present report describes a method for recording total electrical activity from the procerebrum in vivo and demonstrates changes in spontaneous rhythmic oscillations in the procerebrum of the snail Helix in response to olfactory stimulation. Recording of total electrical activity from the snail procerebrum in conditions of free behavior demonstrated a significant reduction in the frequency and an increase in the amplitude of spontaneous oscillations in the procerebrum in the first 10 sec after presentation of the odor of cineole at low concentration, which corresponds to the time required for performance of the olfactory tentacle withdrawal reaction. Presentation of the odor at high concentration resulted in a significant increase in the amplitude of spontaneous oscillations in the procerebrum over the time interval 11-20 sec after odor presentation and a tendency to a decrease in the frequency, which correlates with an increase in the time required for the olfactory tentacle withdrawal reaction. These data suggest an implicit relationship between the amplitude and frequency of oscillations in odor recognition. PMID- 17922242 TI - The contribution of synaptic plasticity in the basal ganglia to the processing of visual information. AB - A mechanism for the involvement of the basal ganglia in the processing of visual information, based on dopamine-dependent modulation of the efficiency of synaptic transmission in interconnected parallel associative and limbic cortex-basal ganglia-thalamus-cortex circuits, is proposed. Each circuit consists of a visual or prefrontal area of the cortex connected with the thalamic nucleus and the corresponding areas in different nuclei of the basal ganglia. The circulation of activity in these circuits is supported by the recurrent arrival of information in the thalamus and cortex. Dopamine released in response to a visual stimulus modulates the efficiencies of "strong" and "weak" corticostriatal inputs in different directions, and the subsequent reorganization of activity in the circuit leads to disinhibition (inhibition) of the activity of those cortical neurons which are "strongly" ("weakly") excited by the visual stimulus simultaneously with dopaminergic cells. The pattern in each cortical area is the neuronal reflection of the properties of the visual stimulus processed by this area. Excitation of dopaminergic cells by the visual stimulus via the superior colliculi requires parallel activation of the disinhibitory input to the superior colliculi via the thalamus and the "direct" pathway" in the basal ganglia. The prefrontal cortex, excited by the visual stimulus via the mediodorsal nucleus of the thalamus, mediates the descending influence on the activity of dopaminergic cells, simultaneously controlling dopamine release in different areas of the striatum and thus facilitating the mutual selection of neural reflections of the individual properties of the visual stimulus and their binding into an integral image. PMID- 17922243 TI - Role of the parietal associative area of the cortex for "counting" behavior in dogs. AB - Experiments were performed on six dogs to study the effects of simultaneous and separate ablation of fields 5 and 7 of the parietal cortex on "counting" behavior. Dogs were trained to discriminate series of five sound clicks presented with variable interstimulus intervals from similar series consisting of three clicks. A food-related operant response (elevation of the right forepaw to place it on the feeder) was used to develop asymmetrical differentiation; the positive signal was a series of five clicks with variable interstimulus intervals and the negative (unreinforced) stimulus was a series of three clicks. Simultaneous bilateral ablation of fields 5 and 7 of the parietal cortex, like bilateral ablation only of field 5, produced profound impairment of differentiation lasting 2-3 months. Isolated bilateral ablation of field 7 produced no impairment of differentiation. These data led to the conclusion that field 5 of the parietal cortex is important for discriminating the numbers of sequential signals. PMID- 17922244 TI - Electrophysiological characteristics of a population of mushroom body neurons in Apis Mellifera honeybees in kynurenine deficiency. AB - Electrophysiological methods were used to study the neurophysiological characteristics of mushroom body neurons in snow (laranja) mutant and wild-type bees. The snow (laranja) mutation, which produces a sharp reduction in the activity of the enzyme tryptophan oxygenase, thus creating a deficiency of all kynurenines in the body, modifies the bioelectrical properties of mushroom body neurons. The parameters most dependent on the snow (laranja) mutation and, thus, the level of endogenous kynurenines, were the duration of action potential afterdepolarization recorded from neurons in the calyx and the amplitude of the postsynaptic potential of neurons in the calyx evoked by focal stimulation of the antennal lobes. There was also a tendency to an increase in the spontaneous spike frequency. These data lead to the conclusion that the effect of the mutation is recessive in nature and appears only in homozygotes (bearing two mutant alleles). PMID- 17922245 TI - NO synthase-dependent increases in extracellular citrulline levels in the nucleus accumbens in an emotional conditioned reflex. AB - Intracerebral microdialysis/HPLC studies in Sprague-Dawley rats showed that the acquisition and execution of an emotional conditioned reflex was accompanied by an increase in the extracellular citrulline level in the nucleus accumbens; citrulline is a co-product of nitric oxide synthesis. The increase in the citrulline level evoked by execution of this reflex decreased after injection of 7-nitroindazole (0.5 mM), a selective inhibitor of neuronal NO synthase, into the nucleus accumbens, and was completely blocked by injection of N-nitroarginine (0.5 mM), a non-selective inhibitor NO synthase. The increase in the nucleus accumbens citrulline level seen during execution of the emotional conditioned reflex was prevented by administration of both of these NO synthase inhibitors. These data suggest that during the acquisition and execution of the emotional conditioned reflex, there is an increase in nitric oxide production in the nucleus accumbens, which arises predominantly as a result of activation of neuronal NO synthase. PMID- 17922246 TI - Interhemisphere differences during tasks involving attention and selection of lateralized stimuli. AB - The state of cortical activation in the parietal and temporal areas of the right and left hemispheres was evaluated using evoked potentials (EP) during tasks consisting of selection of visual stimuli lateralized in the right and left visual fields and needing three different types of attention: to stimulus shape, to stimulus position, and simultaneously to stimulus shape and position. EP were recorded in 15 young healthy experimental subjects using six cortical leads: P3, P4, T3, T4, T5, and T6; the following endogenous EP components (in standard terminology) were analyzed: contingent negative variation (CNV), N1, P3, and the N1-P3 complex. Asymmetry in evoked potentials was assessed in terms of differences to contra-and ipsilateral stimuli in the right and left hemispheres. EP asymmetry was detected in the right hemisphere in all types of selection of lateralized stimuli. The magnitude of asymmetry in the right hemisphere depended on the level (or intensity) of attention: the degree of asymmetry increased with increases in the need for attention to analyze the stimuli. There was a significant relationship between the magnitude of asymmetry and the latent periods of the subjects' responses. The functional significance of these data demonstrating asymmetry may be that it provides better spatial differentiation of visual signals in the right hemisphere, along with dominance of the right hemisphere in attention tasks. PMID- 17922247 TI - Magnesium ions prevent the development of hyperkinesia evoked by administration of picrotoxin into the rat neostriatum. AB - Chronic experiments on rats were performed to study the effects of daily administration of the GABA(A) receptor antagonist picrotoxin (2 microg) into the rostral neostriatum in rats. Picrotoxin was injected in 1 microl of sterile apyrogenic physiological saline or in 1 microl of 1.0 M or 1.5 M MgCl(2); control animals received physiological saline or 1.0 M MgCl(2). Picrotoxin given in physiological saline induced impairments in conditioned reflex (avoidance) and free motor behavior and produced clear stereotypical imperative movements in the form of choreomyoclonic hyperkinesia of the paws, head, and trunk. These motor impairments resembled the manifestaitons of basal ganglia dysfunction typical of Huntingdon's chorea in humans. Magnesium ions prevented both the development of hyperkinesia and impairments of learned behavior. Given that magnesium is a nonspecific calcium channel antagonist, it can be suggested that one of the leading mechanisms of development of hyperkinesias is impairment of calcium homeostasis in striatal neurons. PMID- 17922248 TI - Effects of administration of sodium glutamate during the neonatal period on behavior and blood corticosterone levels in male mice. AB - Treatment of male DBA/2 mice with sodium glutamate (4 mg/g) on postnatal days 1, 3, 5, 7, and 9 induced reductions in the numbers of square crossings, vertical rearings, excursions to the center, and the time spent in the center in adulthood, as compared with a group of males given physiological saline at the same times. These measures showed no change as compared with intact animals. In the light-dark test, the time spent by mice in the light sector was greater after administration of sodium glutamate than after administration of physiological saline but did not differ from that in intact animals. In the acoustic startle reflex test, sodium glutamate decreased startle amplitude but had no effect on the magnitude of prestimulus inhibition. Sexual motivation in males decreased after sodium glutamate, physiological saline producing a tendency to decreased sexual motivation. Neonatal administration of sodium glutamate increased basal blood corticosterone in adult males by a factor of 4, while physiological saline had no effect on this measure. These results lead to the conclusion that neonatal administration of sodium glutamate decreases motor and investigative activity, anxiety, and sexual motivation in adult male mice and increases basal corticosterone. Physiological saline increased all these parameters apart from sexual motivation, though this was not associated with changes in basal corticosterone. PMID- 17922249 TI - Structural basis of the involvement of the striopallidum and pedunculopontine tegmental nucleus in the organization of adaptive behavior. AB - Retrograde axonal transport was used to study the organizational characteristics of the afferent and efferent projection systems of individual substructures of the pedunculopontine tegmental nucleus and the functionally diverse (motor, limbic) areas of structures in the striopallidum. The structural bases of the conduction of functionally diverse information and its integration in the basal ganglia system and tegmental area were evaluated. The morphological data obtained here aid our understanding of the morphofunctional bases of the interaction of these structures and their involvement in adaptive behavior. PMID- 17922250 TI - Involvement of nuclear progesterone receptors in the formation of anxiety in female mice. AB - We report here studies on the delayed effects of exogenous progesterone on the formation of anxiety in female mice. Ovariectomized female mice were given seven days of replacement therapy either with the two main ovarian hormones progesterone and estradiol benzoate-or with progesterone only; levels of anxiety were measured six hours later in the elevated plus-maze. The role of nuclear progesterone receptors in controlling the level of anxiety was assessed by giving some mice injections of the synthetic progesterone receptor blocker mifepristone 2 h before the last dose of hormones. An immunohistochemical method was used to study changes in the number of nuclear progesterone receptors in different areas of the brains of experimental animals. These studies showed that progesterone has a delayed enhancing effect on anxiety in female mice. The role of nuclear progesterone receptors in forming this behavioral characteristic was supported by a strong correlation between changes in the numbers of progesterone receptor immunopositive cells in several brain structures and the level of anxiety. Prior blockade of progesterone receptors using mifepristone led to a maximal reduction in the level of anxiety, which was also evidence for a role for the genomic mechanisms of action of progesterone in controlling anxiety in females. PMID- 17922251 TI - Low cytotoxicity effect of dendrosome as an efficient carrier for rotavirus VP2 gene transferring into a human lung cell line : dendrosome, as a novel intranasally gene porter. AB - The efficiency of dendrosome (a gene porter) was assessed in transferring recombinant human rotavirus VP2 cDNA into A549, a human lung cell line. After gene transferring, transmission electron microscopy showed core-like particles (CLPs) formation in the transfected cells both with dendrosome and lipofectamine porters. In addition, western blotting analysis showed that the expression of VP2 gene was almost equal in the dendrosome and lipofectamine-transfected cells. Also, the cytotoxicity studies revealed that dendrosome had a lower cytotoxicity than lipofectamine. Therefore, our study may introduce dendrosome as a possible carrier for gene transferring into the human lung cell line, especially, for intranasally administration of DNA vaccines. PMID- 17922252 TI - Genetic polymorphism c.1438A>G of the 5-HT(2A) receptor is associated with abdominal obesity in Chinese Northern Han population. AB - OBJECTIVE: We examined the potential impact of the 5-hydroxytryptamine 2A receptor (5-HT(2A)R) c.1438A>G promoter polymorphism on obesity and estimates of insulin, glucose as well as lipid metabolism. METHODS: The genotypes and allelic frequencies of the 5-HT(2A)R c.1438A>G were examined with polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) in 210 patients with overweight/obesity and 216 unrelated healthy subjects. RESULTS: The genotype (AA, AG, and GG) distribution of c.1438A>G polymorphism of the 5-HT(2A)R gene promoter was 35%, 46%, and 19% in patients, and 32%, 56%, 12% in controls, respectively, no significant difference was found between two groups. Association of genetic diversity of 5-HT(2A)R c.1438A>G with the total body fat, fat distribution and clinical characteristics revealed that overweight/obese men carrying G allele were associated with increased body mass index (P = 0.043), waist circumference (P = 0.038), waist-to-hip ratio (P = 0.045), in comparison with patients who carrying A allele, but there were no significant difference between the c.1438A>G genotype groups in overweight/obese women. CONCLUSION: No significant associations were detected. However, the present study suggests the possibility that an abnormal production rate of the 5-HT(2A)R c.1438A>G gene product might lead to the development of abdominal obesity in men but not in women. PMID- 17922253 TI - Molecular and functional analysis of a novel recombinant clone of rat (Rattus norvegicus) CD40 ligand (CD40L) gene. AB - Genetic material obtained from various individuals may contain certain polymorphisms which may conflict with the predetermined DNA sequence and consequently, may modulate the function of gene products. In this study, coding sequence of rat CD40 ligand (CD40L, CD154) was obtained from activated splenocytes, amplified, and cloned into a eukaryotic expression vector by using directional cloning method. Sequence of the recombinant rat CD40L DNA, pCD40L IRES2-EGFP (pCD40L), was compared with the previously reported rat CD40L cDNA sequences and a 99% identity was found. Differing nucleotides were on the positions; 122-T/C, 341-G/A, 476-G/A, 762-T/A. Further alignment analysis showed that pCD40L was collectively carrying the nucleotides each previously reported by different groups. The sequence was submitted to NCBI GenBank and nucleotide database accession number EF066490 was obtained. Following transfection of the construct into NIH/3T3 cell line, novel CD40L clone was functionally expressed de novo, increasing the expression of CD80 and CD86 costimulatory molecules and augmenting the proliferation rate of effector splenocytes in immune reactions ex vivo. Based on these data, here we report a novel recombinant clone of the rat CD40L gene which may represent a potential polymorphic variant. PMID- 17922254 TI - Comparison of the cardiac effects between quinazoline-based alpha1-adrenoceptor antagonists on occlusion-reperfusion injury. AB - Quinazoline-based compounds such as prazosin and its congeners including doxazosin, bunazosin, and terazosin are widely used as antihypertensive agents. However, there were many clinical observations showing that using these agents may result in higher risk of cardiovascular accidents in recent years. In this study, we compared the effects of four alpha-adrenoceptor antagonists: prazosin, doxazosin, bunazosin, and terazosin on occlusion-reperfusion injury. Langendorff perfused rat hearts were pretreated with these four antagonists, and then the left main coronary artery was occluded. After 30 min occlusion, the hearts were reperfused for 2 h and the infarct sizes were measured. Two of the compounds studied, prazosin and doxazosin, apparently increased infarct size, CK-MB, and LDH activities after 2 h reperfusion. In contrast, bunazosin decreased infarct size and those biochemical indicators of cellular damage compared to control hearts. Although infarct size after reperfusion was differently changed by these four alpha-adrenoceptor antagonists, TUNEL-positive nuclei and caspase-3 protein expressions of all the groups were not significantly different. We supposed that the different effects of these four agents on infarct size came from the difference in necrosis rather than apoptosis. PMID- 17922255 TI - Characterization of cocaine-elicited cell vacuolation: the involvement of calcium/calmodulin in organelle deregulation. AB - The sizes of organelles are tightly regulated in the cells. However, little is known on how cells maintain the homeostasis of these intracellular compartments. Using cocaine as a model compound, we have characterized the mechanism of deregulated vacuolation in cultured rat liver epithelial Clone 9 cells. The vacuoles were observed as early as 10 min following cocaine treatment. Removal of cocaine led to vacuole degeneration, indicating vacuolation is a reversible process. The vacuoles could devour intracellular materials and the vacuoles originated from late endosome/lysosome as indicated by immunofluorescence studies. Instant calcium influx and calmodulin were required for the initiation of vacuole formation. The unique properties of these late endosome/lysosome derived vacuoles were further discussed. In summary, cocaine elicited a new type of deregulated vacuole and the involvement of calcium/calmodulin in vacuolation could shed light on prevention or treatment of cocaine-induced cytotoxicity. PMID- 17922256 TI - Assessment of hepatic arterial anatomy in keeping with preservation of the vasculature while performing pancreatoduodenectomy: an opinion. AB - BACKGROUND: Pancreatic surgeons often must make decisions regarding hepatic artery (HA) resection while performing a pancreatoduodenectomy (PD). The purpose of this report was to review and summarize HA resection experience with a focus on vascular preservation during PD and to develop a useful guideline for pancreatic surgeons in dealing with these needs. METHODS: We reviewed 1324 cases that had available computed tomographic and angiographic findings and summarized the problematic HA variations encountered in PD. In reviewing our PD series (n = 254), we have created a set of guidelines that enable a pragmatic approach to the unique variations in HA and the risks of cancer invasion. RESULTS: Challenging HA variations during PD were found in 20.1% of the cases and included the common HA arising from the superior mesenteric artery (SMA) (2.34%), a replaced right HA (RHA) from the SMA (9.82%), an RHA or left HA from the gastroduodenal artery (0.97%), and the right anterior or right posterior HA from the SMA (1.06%), among others. In our PD series, the problematic HAs (15.8%) were preserved, except for a single case (0.4%) in which PD involved en bloc resection of the RHA from the SMA due to a cancerous invasion and without right hemihepatectomy. CONCLUSIONS: Surgeons should have knowledge of the anatomically variable vasculature of the HA when planning for PD. Preoperative imaging studies can aid and should be performed in anticipation of the potential HA variations during PD. PMID- 17922257 TI - Contribution of the BRCA1 and BRCA2 mutations to breast cancer in Tunisia. AB - Hereditary breast cancer accounts for 3-8% of all breast cancers, with mutations in the BRCA1 and BRCA2 genes responsible for up to 30% of these. To investigate the prevalence of BRCA1 and BRCA2 gene mutations in breast cancer patients with affected relatives in Tunisia, we studied 36 patients who had at least one first degree relative with breast and/or ovarian cancer Thirty-four 34 patients were suggestive of the BRCA1 mutation and two were suggestive of the BRCA2 mutation, based on the presence of male breast cancer detected in their corresponding pedigrees. Four mutations in BRCA1 were detected, including a novel frame-shift mutation (c.211dupA) in two unrelated patients and three other frameshift mutations--c.4041delAG, c.2551delG and c.5266dupC. Our study is the first to describe the c.5266dupC mutation in a non-Jewish Ashkenazi population. Two frameshift mutations (c.1309del4 and c.5682insA) were observed in BRCA2. Nineteen percent (7/36) of the familial cases had deleterious mutations of the BRCA1 or BRCA2 genes. Almost all patients with deleterious mutations of BRCA1 reported a family history of breast and/or ovarian cancer in the index case or in their relatives. Our data are the first to contribute to information on the mutation spectrum of BRCA genes in Tunisia, and we give a recommendation for improving clinical genetic testing policy. PMID- 17922258 TI - The high resolution NMR structure of the third SH3 domain of CD2AP. AB - CD2 associated protein (CD2AP) is an adaptor protein that plays an important role in cell to cell union needed for the kidney function. CD2AP interacts, as an adaptor protein, with different natural targets, such as CD2, nefrin, c-Cbl and podocin. These proteins are believed to interact to one of the three SH3 domains that are positioned in the N-terminal region of CD2AP. To understand the network of interactions between the natural targets and the three SH3 domains (SH3-A, B and C), we have started to determine the structures of the individual SH3 domains. Here we present the high-resolution structure of the SH3-C domain derived from NMR data. Full backbone and side-chain assignments were obtained from triple-resonance spectra. The structure was determined from distance restraints derived from high-resolution 600 and 800 MHz NOESY spectra, together with phi and psi torsion angle restraints based on the analysis of 1HN, 15N, 1Halpha, 13Calpha, 13CO and 13Cbeta chemical shifts. Structures were calculated using CYANA and refined in water using RECOORD. The three-dimensional structure of CD2AP SH3-C contains all the features that are typically found in other SH3 domains, including the general binding site for the recognition of polyproline sequences. PMID- 17922259 TI - A global analysis of NMR distance constraints from the PDB. AB - Information obtained from Nuclear Magnetic Resonance (NMR) experiments is encoded as a set of constraint lists when calculating three-dimensional structures for a protein. With the amount of constraint data from the world wide Protein Data Bank (wwPDB) that is now available, it is possible to do a global, large-scale analysis using only information from the constraints, without taking the coordinate information into account. This article describes such an analysis of distance constraints from NOE data based on a set of 1834 NMR PDB entries containing 1909 protein chains. In order to best represent the quality and extent of the data that is currently deposited at the wwPDB, only the original data as deposited by the authors was used, and no attempt was made to 'clean up' and further interpret this information. Because the constraint lists provide a single set of data, and not an ensemble of structural solutions, they are easier to analyse and provide a reduced form of structural information that is relevant for NMR analysis only. The online resource resulting from this analysis ( http://www.ebi.ac.uk/msd/srv/docs/NMR/analysis/results/html/comparison.html ) makes it possible to check, for example, how often a particular contact occurs when assigning NOESY spectra, or to find out whether a particular sequence fragment is likely to be difficult to assign. In this respect it formalises information that scientists with experience in spectrum analysis are aware of but cannot necessarily quantify. The analysis described here illustrates the importance of depositing constraints (and all other possible NMR derived information) along with the structure coordinates, as this type of information can greatly assist the NMR community. PMID- 17922260 TI - The NMR structure of the TRADD death domain, a key protein in the TNF signaling pathway. PMID- 17922261 TI - Global functional analyses of rice promoters by genomics approaches. AB - Promoters play key roles in conferring temporal, spatial, chemical, developmental, or environmental regulation of gene expression. Promoters that are subject to specific regulations are useful for manipulating foreign gene expression in plant cells, tissues, or organs with desirable patterns and under controlled conditions, and have been important for both basic research and applications in agriculture biotechnology. Recent advances in genomics technologies have greatly facilitated identification and study of promoters in a genome scale with high efficiency. Previously we have generated a large T-DNA tagged rice mutant library (TRIM), in which the T-DNA was designed with a gene/promoter trap system, by placing a promoter-less GUS gene next to the right border of T-DNA. GUS activity screens of this library offer in situ and in planta identifications and analyses of promoter activities in their native configurations in the rice genome. In the present study, we systematically performed GUS activity screens of the rice mutant library for genes/promoters constitutively, differentially, or specifically active in vegetative and reproductive tissues. More than 8,200 lines have been screened, and 11% and 22% of them displayed GUS staining in vegetative tissues and in flowers, respectively. Among the vegetative tissue active promoters, the ratio of leaf active versus root active is about 1.6. Interestingly, all the flower active promoters are anther active, but with varied activities in different flower tissues. To identify tissue specific ABA/stress up-regulated promoters, we compared microarray data of ABA/stress induced genes with those of tissue specific expression determined by promoter trap GUS staining. Following this approach, we showed that the peroxidase 1 gene promoter was ABA up-regulated by 4 fold within 1 day of exposure to ABA and its expression is lateral root specific. We suggest that this be an easy bioinformatics approach in identifying tissue/cell type specific promoters that are up-regulated by hormones or other factors. PMID- 17922263 TI - Close association of centrosomes to the distal ends of the microbody during its growth, division and partitioning in the green alga Klebsormidium flaccidum. AB - Division and partitioning of microbodies (peroxisomes) of the green alga Klebsormidium flaccidum, whose cells contain a single microbody, were investigated by electron microscopy. In interphase, the rod-shaped microbody is present between the nucleus and the single chloroplast, oriented perpendicular to the pole-to-pole direction of the future spindle. A centriole pair associates with one distal end of the microbody. In prophase, the microbody changes not only in shape, from a rodlike to a branched form, but also in orientation, from perpendicular to parallel to the future pole-to-pole direction. Duplicated centriole pairs are localized in close proximity to both distal ends of the microbody. In metaphase, the elongated microbody flanks the open spindle, with both distal ends close to the centriole pair at either spindle pole. The microbody further elongates in telophase and divides after septum formation (cytokinesis) has started. The association between the centrioles and both distal ends of the microbody is maintained throughout mitosis, resulting in the distal ends of the elongated microbody being fixed at the cellular poles. This configuration of the microbody may be favorable for faithful transmission of the organelle during cell division. After cytokinesis is completed, the microbody reverts to the perpendicular orientation by changing its shape. Microtubules radiating from the centrosomes flank the side of the microbody throughout mitosis. The close association of centrosomes and microtubules with the microbody is discussed in respect to the partitioning of the microbody in this alga. PMID- 17922264 TI - Hydroquinone peroxidase activity of maize root mitochondria. AB - The oxidation of hydroquinone with H(2)O(2) in the presence of mitochondria isolated from maize (Zea mays L.) roots was studied. The results indicate that a reduced form of quinone may be a substrate of mitochondrial peroxidases. Specific activities in different mitochondrial isolates, the apparent K (m) for hydrogen peroxide and hydroquinone, and the influence of some known peroxidase inhibitors or effectors are presented. Zymographic assays revealed that all mitochondrial peroxidases, which were stained with 4-chloro-1-naphthol, were capable of oxidizing hydroquinone. A possible antioxidative role of hydroquinone peroxidase in H(2)O(2) scavenging within the mitochondria, in cooperation with ascorbate or coupled with mitochondrial NAD(P)H dehydrogenases, is proposed. PMID- 17922265 TI - Possible roles of actin and myosin during anaphase chromosome movements in locust spermatocytes. AB - We tested whether the mechanisms of chromosome movement during anaphase in locust (Locusta migratoria L.) spermatocytes might be similar to those described for crane-fly spermatocytes. Actin and myosin have been implicated in anaphase chromosome movements in crane-fly spermatocytes, as indicated by the effects of inhibitors and by the localisations of actin and myosin in spindles. In this study, we tested whether locust spermatocyte spindles also utilise actin and myosin, and whether actin is involved in microtubule flux. Living locust spermatocytes were treated with inhibitors of actin (latrunculin B and cytochalasin D), myosin (BDM), or myosin phosphorylation (Y-27632 and ML-7). We added drugs (individually) during anaphase. Actin inhibitors alter anaphase: chromosomes either completely stop moving, slow, or sometimes accelerate. The myosin inhibitor, BDM, also alters anaphase: in most cases, the chromosomes drastically slow or stop. ML-7, an inhibitor of MLCK, causes chromosomes to stop, slow, or sometimes accelerate, similar to actin inhibitors. Y-27632, an inhibitor of Rho-kinase, drastically slows or stops anaphase chromosome movements. The effects of the drugs on anaphase movement are reversible: most of the half bivalents resumed movement at normal speed after these drugs were washed out. Actin and myosin were present in the spindles in locations consistent with their possible involvement in force production. Microtubule flux along kinetochore fibres is an actin-dependent process, since LatB completely removes or drastically reduces the gap in microtubule acetylation at the kinetochore. These results suggest that actin and myosin are involved in anaphase chromosome movements in locust spermatocytes. PMID- 17922267 TI - Cell wall sheath surrounding calcium oxalate crystals in mulberry idioblasts. AB - The distribution and ultrastructural features of idioblasts containing calcium oxalate crystals were studied in leaf tissues of mulberry, Morus alba L. In addition to the calcium carbonate crystals formed in epidermal idioblasts, large calcium oxalate crystals were deposited in cells adjacent to the veins and surrounded by a cell wall sheath which had immunoreactivity with an antibody recognizing a xyloglucan epitope. The wall sheath formation indicates exclusion of the mature crystal from the protoplast. PMID- 17922266 TI - Orientation of microtubules suggests a role in mRNA transportation in fertilized eggs of Chinese pine (Pinus tabulaeformis). AB - Polysomes become associated with microtubules (MTs) in egg cells of Chinese pine upon fertilization, providing direct evidence for MT-based intracellular mRNA and polysome localization. We have investigated by immunoelectron microscopy the orientation and spatial distribution of MTs and their association with polysomes in the fertilized egg cells. There is a perinuclear accumulation of MTs and polysomes in the zygote soon after fertilization. At this time, some of the MTs are perpendicular to the nuclear envelope and directly connected to the outer membrane or nuclear-pore complexes (NPC) at one end, and the other ends reach to the outer tier or cortical MTs that are parallel to the long axis of the zygote. The polysomes in the perinuclear region show the same spatial and temporal pattern as the MTs. Immunolocalization of the mRNA-binding protein hnRNP indicates that the mRNAs are loaded onto the nucleus-associated MTs immediately after their export from the nuclear-pore complexes. The polysomes and mRNAs are then transported from these MTs to the outer tier and/or cortical MTs, where they further localize to the polar region of the cell. PMID- 17922268 TI - Flow cytometric identification of two different rhodamine-123-stained mitochondrial populations in maize leaves. AB - Flow cytometric analysis of mitochondria isolated from maize leaves revealed two distinct rhodamine-123-stained fluorescence populations distinguishable by their main fluorescence channel. Further microscopic observation of mitochondria stained with Janus Green B and rhodamine-123 revealed the occurrence of differently sized mitochondrial particles. It was shown by pulsed-field gel electrophoresis that the DNA from the isolated mitochondria ranged in size from 45 to 100 kb. These results suggest that different types of mitochondria with different physiological status, mass, and genomic DNA size probably coexist and carry out different physiological functions throughout the whole process of maize leaf growth and development. PMID- 17922269 TI - Utility of the "buddy" wire in intracranial procedures. AB - Patients undergoing neurointerventional procedures with excessively tortuous vascular anatomy often have limited treatment options. The ability to pass and maintain the stability of micro-guidewires, catheters, and interventional devices is often a product of guide catheter steadiness. A companion wire passed through the lumen of the guide catheter to increase the guide catheter's stiffness can overcome the challenges associated with tortuous anatomy; this companion wire is referred to as a "buddy" wire. We demonstrate the technical success of this system by presenting a patient whose endovascular treatment would have otherwise been impossible without a "buddy" wire. PMID- 17922270 TI - Mucoepidermoid carcinoma of bronchus in a pediatric patient: (18)F-FDG PET findings. AB - In children, primary neoplasms of the tracheobronchial tree and lungs are rare; most are malignant. Of the primary malignant pulmonary neoplasms arising in childhood, mucoepidermoid carcinoma accounts for approximately 10%. Due to its well-confined local growth within the airway, mucoepidermoid carcinoma commonly produces respiratory symptoms from progressive tracheal or bronchial obstruction. Mucoepidermoid tumor has minimal metastatic potential in children, and local resection alone is the current treatment of choice. Early detection, diagnosis, and surgical resection of mucoepidermoid tumor are especially important in pediatric patients since the bulk of the remaining pulmonary parenchyma can be preserved, thereby decreasing the thoracic deformity and pulmonary functional morbidity. Radiographic and CT imaging findings of bronchial mucoepidermoid carcinoma in children have been described in several case reports. However, to the best of our knowledge, imaging findings of 2-((18)F)-fluoro-2-deoxy-D: glucose positron emission tomography ((18)F-FDG PET) of mucoepidermoid carcinoma of the bronchus in pediatric patients have not been well established. We report a mucoepidermoid carcinoma arising from the right upper lobe bronchus in a 15-year old girl with an emphasis on the (18)F-FDG PET findings. PMID- 17922271 TI - Comparison of SSS and SRS calculated from normal databases provided by QPS and 4D MSPECT manufacturers and from identical institutional normals. AB - PURPOSE: There is proven evidence for the importance of myocardial perfusion single-photon emission computed tomography (SPECT) with computerised determination of summed stress and rest scores (SSS/SRS) for the diagnosis of coronary artery disease (CAD). SSS and SRS can thereby be calculated semi quantitatively using a 20-segment model by comparing tracer-uptake with values from normal databases (NDB). Four severity-degrees for SSS and SRS are normally used: <4, 4-8, 9-13, and > or =14. Manufacturers' NDBs (M-NDBs) often do not fit the institutional (I) settings. Therefore, this study compared SSS and SRS obtained with the algorithms Quantitative Perfusion SPECT (QPS) and 4D-MSPECT using M-NDB and I-NDB. METHODS: I-NDBs were obtained using QPS and 4D-MSPECT from exercise stress data (450 MBq (99m)Tc-tetrofosmin, triple-head-camera, 30 s/view, 20 views/head) from 36 men with a low post-stress test CAD probability and visually normal SPECT findings. Patient group was 60 men showing the entire CAD spectrum referred for routine perfusion-SPECT. Stress/rest results of automatic quantification of the 60 patients were compared to M-NDB and I-NDB. After reclassifying SSS/SRS into the four severity degrees, kappa values were calculated to objectify agreement. RESULTS: Mean values (vs M-NDB) were 9.4 +/- 10.3 (SSS) and 5.8 +/- 9.7 (SRS) for QPS and 8.2 +/- 8.7 (SSS) and 6.2 +/- 7.8 (SRS) for 4D-MSPECT. Thirty seven of sixty SSS classifications (kappa = 0.462) and 40/60 SRS classifications (kappa = 0.457) agreed. Compared to I-NDB, mean values were 10.2 +/- 11.6 (SSS) and 6.5 +/- 10.4 (SRS) for QPS and 9.2 +/- 9.3 (SSS) and 7.2 +/- 8.6 (SRS) for 4D-MSPECT. Forty four of sixty patients agreed in SSS and SRS (kappa = 0.621 resp. 0.58). CONCLUSION: Considerable differences between SSS/SRS obtained with QPS and 4D-MSPECT were found when using M-NDB. Even using identical patients and identical I-NDB, the algorithms still gave substantial different results. PMID- 17922272 TI - Topotecan and carboplatin in patients with platinum-sensitive recurrent ovarian cancer. Results of a multicenter NOGGO: phase I/II study. AB - OBJECTIVE: Second-line treatment with paclitaxel and carboplatin enhances survival of women with platinum-sensitive recurrent ovarian cancer (ROC). However, because of its cumulative neurotoxicity, there is a strong demand for platinum-combinations with better therapeutic index. Because of its pharmacological properties, topotecan is a good adjunct to carboplatin in this setting, but its safety and efficacy remains to be defined. METHODS: Patients with platinum-sensitive ROC were eligible in this multicenter phase I/II study, stratified according to treatment-free interval (TFI). Dose level 0 consisted of topotecan 1 mg/m(2)/d1-3/q21d plus carboplatin AUC5/d3/q21d. DLT was defined as grade > or =3 neutropenia or thrombocytopenia or grade > or =3 non-hematological toxicity excluding alopecia, nausea and vomiting, accompanied by a treatment delay >1 week. RESULTS: From June 2004 to August 2005, 26 patients were enrolled, receiving a total of 145 cycles of chemotherapy. MTD was reached at topotecan 0.75 mg/m(2) and carboplatin AUC5. We observed a single grade 4 leucopenia. There were 3 (12%), 15 (58%) and 8 (31%) events of grade 3/4 hematological anaemia, leucopenia, and thrombocytopenia. Response rate was 67% (95% CI 43-85), median progression-free survival 9.5 months (95% CI 7.3-12.0), median overall survival 19.4 months (95% CI 12.3-26.9). None of the toxicity or efficacy endpoints were associated with TFI. CONCLUSION: Topotecan and carboplatin is a well tolerated novel doublet option for women with platinum sensitive ROC. We encourage further studies on this approach, but to limit the doses of topotecan to 0.75 mg/m(2)/d1 3 and carboplatin AUC 5/d3. PMID- 17922273 TI - Effect of medical castration on CYP3A4 enzyme activity using the erythromycin breath test. AB - PURPOSE: Testosterone administration can lead to increased antipyrine clearance in humans. Medical or surgical castration is a standard treatment of progressive prostate carcinoma, but the effect of the subsequent fall of testosterone concentrations upon drug metabolism has not been reported. METHODS: Eleven men with a biopsy-proven diagnosis of progressive prostate cancer were enrolled after providing informed consent. CYP3A4 activity was determined using the erythromycin breath test (EBT) in each patient prior to their beginning with an LHRH-agonist (leuprolide or goserelin). No patients had elected to undergo orchiectomy during the period of subject accrual. Each subject underwent a second EBT 2 months after beginning LHRH suppression. Blood samples were collected at these time points to determine changes in testosterone and leutinizing hormone. RESULTS: All subjects had a predictable drop in serum testosterone concentrations over the 8-week course of the study, but concentrations in three did not fall below castrate levels (<50 ng/dl). There was no statistically significant change in CYP3A4 activity using the EBT method (p = 0.88). The extent and direction of changes in CYP3A4 activity was highly variable, with three subjects experiencing an increase in activity, and five demonstrating a decrease in activity. CONCLUSION: There is no clinically significant change in CYP3A4 activity after medical castration. No changes in the clearance of docetaxel or other CYP3A4 substrates are likely during and after medical castration. Although similar findings are expected after orchiectomy, we were not able to test this presumption because of patient preference for medical castration. PMID- 17922274 TI - Irinotecan plus cisplatin and dexamethasone (ICD) combination chemotherapy for patients with diffuse large B-cell lymphoma previously treated with Rituximab plus CHOP. AB - PURPOSE: The therapeutic strategy for relapsed or refractory patients with diffuse large B-cell lymphoma (DLBL) remains challenging yet. Salvage therapy has been tried for these patients according to their clinical status. We studied ICD (irinotecan, cisplatin and dexamethasone) regimen as salvage chemotherapy for DLBL patients previously treated with RCHOP. METHODS: Between February 2005 and May 2006, 15 patients were treated prospectively with ICD chemotherapy; irinotecan 65 mg/m(2)/day on days 1 and 8, cisplatin 30 mg/m(2)/day on days 1 and 8, and dexamethasone 40 mg/day on days 1-2 and 8-9. This schedule was planned to repeat every 3 weeks until disease progression, severe toxicity or stem cell transplantation. RESULTS: Of the 14 patients evaluable for response, 3 patients achieved CR, 7 patients PR, with 1 SD and 3 PD; overall response rate 71% (10/14; 95% confidence interval, 47-95%). The median progression free survival (PFS) and event free survival (EFS) were 113 (range 21-493+) and 77 (range 21-324+) days, respectively. The median overall survival was 267 (range 31-493+) days. Grade 3/4 neutropenia and grade 3 neutropenic fever were observed in 67% (22/33) and 18% (6/33) of cycles, respectively. There were 20% of grade 3/4 nausea and diarrhea observed. CONCLUSIONS: The ICD regimen with current schedule showed high response rate for DLBL patients previously treated with RCHOP. But the high incidence of neutropenia led to delay of subsequent cycles causing dose intensity reduced, which seems to be related with short PFS and EFS. PMID- 17922275 TI - Liposomal pegylated doxorubicin plus vinorelbine combination as first-line chemotherapy for metastatic breast cancer in elderly women > or = 65 years of age. AB - PURPOSE: No standard chemotherapy has been so far definitely settled for elderly patients with metastatic breast cancer (MBC). In order to identify a regimen with acceptable efficacy and low burden of non-overlapping toxic effects, a combination consisting of liposomal pegilated doxorubicin (PLD) with alternating oral and intravenous vinorelbine (NVB) has been investigated in a phase II study. METHODS: Thirty-four consecutive patients (median age 71 years; range 65-82) with MBC have been enrolled. Based on 4-weekly cycles, PLD 40 mg/m(2) plus NVB 25 mg/m(2) i.v., have been administered intravenously on day 1 and oral NVB 60 mg/m(2) on day 15. RESULTS: All patients were assessable for safety and efficacy. In all, 17 responses were documented with three complete responses (CR) and 14 partial responses, with an overall response rate of 50% (95% CI 36-66). Median overall survival time was 13 months and the median time to progression 8 months. Interestingly, all the patients with CR are still alive with a disease-free survival of more than 1 year. The main toxicity was neutropenia: grade 3 in 15% and grade 4 in 11% of patients, respectively. Febrile neutropenia was recorded in three patients not requiring dose reduction. Other frequently reported adverse events included: anemia, nausea, vomiting, stomatitis, all rarely severe. The evaluation of quality of life (QoL) did not show any significant change during the study. CONCLUSIONS: Our data suggest that this combination is active and well tolerated in elderly patients with MBC and could represent another efficacious chance for the management of this population. PMID- 17922276 TI - Safety evaluation of oral fluoropyrimidine S-1 for short- and long-term delivery in advanced gastric cancer: analysis of 3,758 patients. AB - PURPOSE: To evaluate the comprehensive safety profile of S-1, a promising novel oral fluoropyrimidine derivative, based on large cohort data. PATIENTS AND METHODS: Study subjects were identified from a prospective registry of 3,758 advanced gastric cancer patients in Japan. Each patient was treated with an identical regimen of S-1 monotherapy (40 mg b.i.d. on days 1-28, every 6 weeks) and assessed for all adverse events. RESULTS: The median duration of treatment was 88 days; 1,605 (43%) patients underwent three or more treatment cycles. The relative dose intensity was 0.87 in the first two cycles (short-term treatment period) and 0.89 thereafter (long-term treatment period). Neutropenia was the most common severe (grade 3-4) hematological event (6.3% in the short-term period and 5.3% in the long-term period). Other hematological or key gastrointestinal events (diarrhea, nausea/vomiting, and stomatitis) had a low incidence of severe cases throughout the whole administration period (0.3-3.8%). The time to onset of severe events did not differ between patients with mild renal impairment (creatinine clearance, 50-79 ml/min) and those with normal renal function (>or=80 ml/min) (hazard ratio, 1.04; 95% CI, 0.87-1.23; P = 0.691). CONCLUSIONS: S-1 had manageable severe toxicity and allowed good compliance regardless of treatment duration. Prolonged administration of the drug was sustainable. PMID- 17922277 TI - Semimechanistic pharmacokinetic/pharmacodynamic model for hepatoprotective effect of dexamethasone on transient transaminitis after trabectedin (ET-743) treatment. AB - PURPOSE: Reversible transient elevations in transaminases have been observed after trabectedin administration. A semimechanistic pharmacokinetic and pharmacodynamic (PKPD) model was developed to evaluate the time course of alanine aminotransferase (ALT) elevation, tolerance development, and the hepatoprotective effect of dexamethasone on trabectedin-induced transient transaminitis following different dosing schedules in cancer patients. PATIENTS AND METHODS: Trabectedin was administered to 711 patients as monotherapy (dose range: 0.024-1.8 mg/m(2)) as 1-, 3-, or 24-h infusions every 21 days; 1- or 3-h infusions on days 1, 8, and 15 every 28 days; or 1-h infusions daily for five consecutive days every 21 days. Population PKPD modeling was performed with covariate evaluation [dexamethasone use (469/711 pt), ECOG performance status scores (89.7% pts or=3 toxicity by 13 and 39% following two and four cycles of therapy, respectively. CONCLUSIONS: A PKPD model quantifying the hepatoprotective effect of dexamethasone on transient and reversible transaminitis following trabectedin treatment has been developed. The model predicts that co-administration of dexamethasone and the suggested dose reduction strategy based on the serum concentration of liver enzymes will enhance the safe use of trabectedin in the clinic. PMID- 17922279 TI - Contrast-enhanced MR imaging of the brain using T1-weighted FLAIR with BLADE compared with a conventional spin-echo sequence. AB - The BLADE and PROPELLER (periodically rotated overlapping parallel lines with enhanced reconstruction) techniques have been proposed to reduce the effect of head motion. Preliminary results have shown that BLADE also reduces pulsation artifacts from venous sinuses. The purpose of this study was to compare T1 weighted FLAIR acquired with BLADE (T1W-FLAIR BLADE) and T1-weighted spin-echo (T1W-SE) for the detection of contrast enhancement in a phantom and in patients with suspected brain lesions and to compare the degree of flow-related artifacts in the patients. A phantom filled with diluted Gd-DTPA was scanned in addition to 27 patients. In the phantom study, the peak contrast-to-noise ratio of T1W-FLAIR BLADE was larger than that of T1W-SE, and the position of the peak was shifted to a lower concentration. In patients, the degree of flow-related artifacts was significantly higher in T1W-SE. Among the 27 patients, 9 had metastatic tumor, and 18 did not. On a patient-by-patient basis, the sensitivity and specificity for the detection of metastatic lesions on axial T1W-SE were 100% and 55.6% respectively, while on axial T1W-FLAIR BLADE they were 100% and 100%. T1W-FLAIR BLADE seems to be capable of replacing T1W-SE, at least for axial post-contrast imaging to detect brain metastases. PMID- 17922280 TI - Aneurysmal bone cyst of spine: a review of literature. AB - Aneurysmal bone cysts (ABC) are benign, highly vascular osseous lesions characterized by cystic, blood-filled spaces surrounded by thin perimeters of expanded bone. Children and young adults are most affected by spinal ABCs. We document a review of literature. PMID- 17922281 TI - Inferior dislocation of the proximal tibiofibular joint: a new type of dislocation with poor prognosis. AB - Dislocation of the proximal tibiofibular joint is an unusual injury. We report a patient, who developed inferior proximal tibiofibular dislocation after a severe motorcycle accident. The dislocation was associated with avulsion of the leg, fractures of the fibula and the ankle and neurovascular lesions. The patient was surgically treated and had a good final outcome. Classifications of proximal tibiofibular dislocations did not include inferior dislocation. This type is always associated with avulsion mechanism and has the poorest prognosis. PMID- 17922278 TI - Cell therapy in myocardial infarction: emphasis on the role of MRI. AB - Despite tremendous progress in myocardial infarct (MI) treatment, mortality rates remain substantial. Permanent loss of cardiomyocytes after ischemic injury, results in irreversible loss of myocardial contractility, reduction in ventricular performance, and may initiate the development of dilated heart failure. The discovery that pluripotent progenitor cells bear the capacity to differentiate to mature cardiac cells raised the hope of cell-based regenerative medicine. Engraftment of stem cells in the damaged myocardium, repair and functional improvement appeared suddenly a nearby reality. Promising results in animal models, and preliminary studies reporting the feasibility and safety of adult stem cell therapy in MI patients led to the first double-blinded randomized, placebo-controlled trials. The initial great enthusiasm for this paradigm shift in MI treatment has been tempered by the mainly negative or modestly positive study findings. Before new, larger clinical trials can be initiated, a number of critical questions and issues need to be considered starting with a scrutinized analysis of currently available data to extending our knowledge of the mechanism of scarless myocardial regeneration. Cardiac cell therapy necessitates a multidisciplinary approach, whereby imaging, in particular MRI, and the input of the imaging specialist is crucial to the success of cardiac cell regenerative medicine. MRI is an appealing technique for cell trafficking depicting engraftment, differentiation and survival. Endomyocardial cell administration can be achieved safely with MR fluoroscopy and MRI is without any doubt the most accurate and reproducible technique to measure study end-points. PMID- 17922282 TI - Prosthetic joint replacement for long bone metastases: analysis of 154 cases. AB - INTRODUCTION: Metastatic bone disease is the most common cause of malignancies to the skeleton in adults. The treatment of bone metastases is frequently palliative aiming to achieve a satisfactory control of pain and to prevent or to treat pathological fractures. In selected cases the resection of a single bone metastasis may improve the survival of the patients. Our experience with bone metastases located in the appendicular skeleton, between 1992 and 2004, is retrospectively reviewed here. MATERIALS AND METHODS: We report a series of 154 patients (95 females and 59 males) treated with prosthesis for metastatic bone disease. Lower limb localization was more frequent with 117 cases, while upper limb was affected in 37 cases. Metastatic breast and renal carcinoma predominated and accounted for 66% of the lesions. Indications to surgery were reported, oncologic outcome was evaluated and functional results were obtained by the Musculoskeletal Tumor Society scoring system. RESULTS: Follow up ranged from 6 months to 12 years (median 26 months). One-year survival was 69.5%, 2-years survival was 44.8%, 5-years survival was 19.5%; and 5 (3.2%) died in the early post surgical period. Functional results were good or higher in 73.8% of patients for the proximal femur, in 50% of patients for the knee and 30.6% of patients for the proximal humerus. CONCLUSION: In this series, satisfactory results were achieved with few complications. We emphasized the importance of giving the patient a definitive treatment and preventing pathological fractures as they determine disability and a spreading of the tumor in the soft tissues, leading to an increased probability of local recurrence. Prosthetic replacement contributes to an improved quality of life and limb functionality and, in selected cases; this radical surgical approach is indicated as it may improve patient's life expectancy. PMID- 17922283 TI - Extreme complications of Fixion nail in treatment of long bone fractures. AB - The authors present their experience related to extreme complications in treatment of diaphyseal fractures of the long bones with application of the Fixion expansion intramedullary nail in a total of 48 surgeries. We have encountered six (12.5%) extreme complications in the management of fractures of 3 humeral, 2 tibial, and 1 femoral bones during its application procedure and postoperative follow-up. Of six cases, two with humeral and tibial fractures developed nonunion and rotational instability because of failure of inflation of the Fixion nail. One of the Fixion nail in humerus was broken spontaneously, and one of the Fixion nail deflated at the follow-up and pseudoarthrosis developed in this patient. In a patient with osteogenesis imperfecta, during the inflation of the nail for the treatment of femur fracture, a new longitudinal fracture occurred and conventional non-locking intramedullary nail was inserted. In a patient with a tibia fracture that was treated with the Fixion nail, new fracture occurred due to its bending after weight bearing in the postoperative period. The Fixion nail application is a new technique for the intramedullar fixation of long bones. It is considered as an effective method for the selective fracture types of long bones. Application may need special training. Since the Fixion has not got rotational stability and rigidity as conventional nailing systems, bending and breaking of the nail may occur during postoperative period in patients with over obesity and hyperactivity. In patients with osteogenesis imperfecta, it may not be the first choice as a nailing system. PMID- 17922284 TI - The safe distance for the superior gluteal nerve in direct lateral approach to the hip and its relation with the femoral length: a cadaver study. AB - INTRODUCTION: The most inferior branch (MIB) of the superior gluteal nerve (SGN) is vulnerable during direct lateral approach to the hip. A safe distance proximal to the tip of the greater trochanter varying from 3 to 5 cm has been reported in different studies. Anatomical studies defining safe zones and clinical studies reporting the results use various reference points, and the oblique course of the MIB contributes to the confusion. Numerous efforts have been made to standardize the safe zone using patient characteristics such as body height; however, contradictory results have been reported. The purpose of this study was to measure the safe distance in line to the gluteal split and also to determine the relationship of the safe distance with femoral length, as a stable component of body height. MATERIALS AND METHODS: Fifteen lower extremities of 12 formalin fixed cadavers (M/F: 7/5) were dissected. The most prominent lateral palpable part of the trochanter major (TM) was determined and the dissection in the gluteus medius muscle (GMM) was performed starting from this point upwards in line of the muscle fibers. The distances between the MIB in the plane of dissection in the GMM to the TM and also to the trochanteric apex (TA) were measured. Femoral lengths were measured between the TM point and the lateral epicondyle. Spearman's correlation and Mann-Whitney U tests were used for statistical analysis. RESULTS: The SGN in 13 hips had spray pattern and neural trunk pattern in two. The plane of dissection was within the anterior third of the GMM in all hips. The average femoral length was 37.5 cm. Average distance between TM and MIB was 44 mm; in three hips, the distance was <30 mm. The average distance between TA and TM was 21 mm. There was no statistically significant correlation between femoral length and TM-MIB distance. CONCLUSION: The distance from the TM to the MIB is highly variable and independent from body height or femoral length. The so called "safe zone" in which damage of significant nerve damage is excluded can have a rather small dimension in some patients. Short patients are not at increased risk and tall patients are not risk free. Modern techniques in total hip replacement which try to minimize proximal interruption of the GMM are therefore justified. PMID- 17922285 TI - Epidemiological characteristics in women with and without endometriosis in the Yale series. AB - OBJECTIVE: The association between demographic factors, menstrual and reproductive characteristics, and clinical profile for women with endometriosis was analyzed in a retrospective case-control study. METHODS: Over a 6-year period, 535 women with endometriosis and 200 infertile women without endometriosis, studied by laparoscopy or laparotomy, were evaluated. Information was then collected in a uniform manner from the patients' medical records. Statistical methods included chi(2) and Mann-Whitney U test. RESULTS: The factors associated with an increased risk for endometriosis include lower body weight, alcohol use (chi(2) = 8.8; P < 0.003), early menarche (chi(2) = 5.08; P < 0.024), shorter cycle length (chi(2) = 13.06; P < 0.001), and heavier menstrual cycles. Pelvic pain was present in 79.1% of women with endometriosis, dysmenorrhea in 70.2%, and dyspareunia in 49.5%. These symptoms were statistically significantly higher in comparison with the infertile women without endometriosis (P < 0.001). Moreover, we found that women with endometriosis had fewer prior pregnancies, elective abortions and ectopic pregnancies compared to women seeking care for infertility, who did not have endometriosis. Interestingly, women with endometriosis were significantly more likely to report a family history of cancer compared to women in control group (chi(2) = 78.2; P < 0.001). CONCLUSIONS: Body habitus, personal habits and menstrual characteristics are all strongly associated with the development of endometriosis. There may also be an association between family history of cancer and the development of endometriosis. PMID- 17922286 TI - Delivery mode and maternal rehospitalization. AB - The rates of rehospitalization after cesarean section (1.41%) are significantly higher than those following spontaneous vaginal delivery (0.33%), mainly due to infection following cesarean section and late bleeding after vaginal delivery. PMID- 17922287 TI - Acoustic analysis of voice using WPCVox: a comparative study with Multi Dimensional Voice Program. AB - In this study, two different tools developed for the parametric extraction and acoustic analysis of voice samples are compared. The main goal of the paper is to contrast the results obtained using the classical Multi Dimensional Voice Program (MDVP), with the results obtained with the novel WPCVox. The aim of this comparison was to find differences and similarities in the parameters extracted with both systems in order to make comparison of measurements and data transfer among both equipments. The study was carried out in two stages: in the first, a wide sample of healthy voices belonging to Spanish-speaking adults from both genders were used to carry out a direct comparison between the results given by MDVP and those obtained with WPCVox. In the second stage, a sample of 200 speakers (53 normal and 173 pathological) taken from a commercially available database of voice disorders were used to demonstrate the usefulness of WPCVox for the acoustic analysis and the characterization of normal and pathological voices. The results conclude that WPCVox provides very reliable measurements which are very similar to those obtained using MDVP, and very similar capabilities to discriminate among normal and pathological voices. PMID- 17922288 TI - A novel plastidial lipoxygenase of maize (Zea mays) ZmLOX6 encodes for a fatty acid hydroperoxide lyase and is uniquely regulated by phytohormones and pathogen infection. AB - Lipoxygenases (LOXs) are members of a large enzyme family that catalyze oxygenation of free polyunsaturated fatty acids into diverse hydroperoxide compounds, collectively called oxylipins. Although LOXs have been well studied in dicot species, reports of the genes encoding these enzymes are scarce for monocots, especially maize. Herein, we reported the cloning, characterization and molecular functional analysis of a novel maize LOX gene, ZmLOX6. The ZmLOX6 nucleotide sequence encodes a deduced translation product of 892 amino acids. Phylogenetic analysis showed that ZmLOX6 is distantly related to previously reported 9- or 13-LOXs from maize and other plant species, including rice and Arabidopsis. Although sequence prediction suggested cytoplasmic localization of this protein, ZmLOX6 protein has been reportedly isolated from mesophyll cell chloroplasts, emphasizing the unique features of this protein. Plastidial localization was confirmed by chloroplast uptake experiments with the in vitro translated protein. Analysis of recombinant protein revealed that ZmLOX6 has lost fatty acid hydroperoxide forming activity but 13-LOX-derived fatty acid hydroperoxides were cleaved into odd-chain omega-oxo fatty acids and as yet not identified C5-compound. In line with its reported abundance in mesophyll cells, ZmLOX6 was predominantly expressed in leaf tissue. Northern blot analysis demonstrated that ZmLOX6 was induced by jasmonic acid, but repressed by abscisic acid, salicylic acid and ethylene and was not responsive to wounding or insects. Further, this gene was strongly induced by the fungal pathogen Cochliobolus carbonum during compatible interactions, suggesting that ZmLOX6 may contribute to susceptibility to this pathogen. The potential involvement of ZmLOX6 in maize interactions with pathogens is discussed. PMID- 17922290 TI - Kaposi's sarcoma after liver transplantation. AB - INTRODUCTION: Kaposi's Sarcoma (KS) is a malignant neoplasm arising from endothelial cells. HHV8-infection represents a key pathogenic determinant for the development of KS. There are no standard criteria to treat KS in immunosuppressed individuals. Six cases (2.1%) of KS occurred in our Center among 285-recipients who underwent liver transplantation (LT) between October 2000 and November 2006. METHODS: Patients were four males and two females. Mean age was 57 years (range 44-65). Indication for LT was ESLD associated/non-associated with hepatocellular carcinoma (HCC). The immunosuppressive regimen consisted of cyclosporine/tacrolimus associated with steroids or daclizumab. HHV8-detection was performed by the serological method before LT, and by polymerase chain reaction (PCR)-analysis after KS. RESULTS: One patient had HCV-related cirrhosis and coinfection from HIV, three had HBV-related cirrhosis, two of these with coexistent HCC. The last two patients had alcoholic-cirrhosis, one with coexistent HCC. Mean time from transplantation to KS was 6.2 months (range 3.8 8.8). Three patients were treated with doxorubicin and three with switch from calcineurin-inhibitors to sirolimus. Three patients expired after 11.5, 8.8, and 7.4 months from KS diagnosis. DISCUSSION: KS should be treated by a multidisciplinary approach to obtain an early diagnosis and best management. Effective treatment with immunosuppression reduction or switch to sirolimus is mandatory and can induce complete regression. PMID- 17922289 TI - Constitutive and herbivore-inducible glucosinolate concentrations in oilseed rape (Brassica napus) leaves are not affected by Bt Cry1Ac insertion but change under elevated atmospheric CO2 and O3. AB - Glucosinolates are plant secondary compounds involved in direct chemical defence by cruciferous plants against herbivores. The glucosinolate profile can be affected by abiotic and biotic environmental stimuli. We studied changes in glucosinolate patterns in leaves of non-transgenic oilseed rape (Brassica napus ssp. oleifera) under elevated atmospheric CO2 or ozone (O3) concentrations and compared them with those from transgenic for herbivore-resistance (Bacillus thuringiensis Cry1Ac endotoxin), to assess herbivory dynamics. Both elevated CO2 and O3 levels decreased indolic glucosinolate concentrations in transgenic and non-transgenic lines, whereas O3 specifically increased the concentration of an aromatic glucosinolate, 2-phenylethylglucosinolate. The herbivore-inducible indolic glucosinolate response was reduced in elevated O3 whereas elevated CO2 altered the induction dynamics of indolic and aliphatic glucosinolates. Herbivore resistant Bt plants experienced minimal leaf damage after target herbivore Plutella xylostella feeding, but exhibited comparatively similar increase in glucosinolate concentrations after herbivory as non-transgenic plants, indicating that the endogenous glucosinolate defence was not severely compromised by transgenic modifications. The observed differences in constitutive and inducible glucosinolate concentrations of oilseed rape under elevated atmospheric CO2 and O3 might have implications for plant-herbivore interactions in Brassica crop ecosystems in future climate scenarios. PMID- 17922291 TI - Overexpression of cFLIP in head and neck squamous cell carcinoma and its clinicopathologic correlations. AB - PURPOSE: The aim of this study was to determine the expression of cellular FLICE like inhibitory protein (cFLIP) in head and neck squamous cell carcinoma (HNSCC) and revealed its possible correlation to Fas protein and tumour clinical parameters. METHODS: The expression of cFLIP was analysed in 58 HNSCC samples and 30 morphologically normal tissues adjacent to the carcinomas using immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis. Furthermore, its possible correlation to the expression of Fas protein and tumour clinicopathologic parameters were discussed. RESULTS: Cellular FLICE-like inhibitory protein was demonstrated to be up regulated in most HNSCC than in normal tissues by immunohistochemistry (p<0.01). Although the mRNA levels of both isoforms of cFLIP, long form (cFLIP(L)) and short form (cFLIP(S)), in HNSCC were higher than those in normal tissues (p<0.01), only cFLIP(L) protein could be detected by western blot. Furthermore, the expression of cFLIP(L) protein was significantly associated with tumour clinical stage (p<0.01) and lymph node metastasis (p=0.01). Since all of the tumours with Fas immunostaining also express cFLIP protein, there was no significant correlation between them (p>0.05). CONCLUSIONS: Overexpression of cFLIP(L) is a frequent event in HNSCC and HNSCC cells in vivo may need it to evade apoptosis mediated by Fas or other receptors, which might contribute to tumour development and progression. PMID- 17922294 TI - Correlation between cystatin C- and renal scan-determined glomerular filtration rate in children with spina bifida. AB - We report on the relationships between serum cystatin C level, glomerular filtration rate (GFR) estimated from a cystatin C-based prediction equation (that of Filler and Lepage), GFR calculated by the Schwartz formula and technetium 99m diethylene triamine penta-acetic acid ((99)Tc-DTPA)-determined GFR in 28 children with spina bifida. All children underwent measurement of height, weight, serum cystatin C level, and serum creatinine level at the time of their renal scan. The relationship between variables was assessed by Pearson correlation. Pearson correlation for the relationship between (99)Tc-DTPA GFR and GFR calculated by the cystatin C-based equation was significant and higher than that of the relationship between (99)Tc-DTPA GFR and GFR calculated by the Schwartz equation, which was not statistically significant. The correlation for Filler GFR was 0.42 (P = 0.03) and for Schwartz GFR was 0.21 (P = 0.28). Although we use renal scan determination of GFR as the best measure, and a creatinine-based formula as the most practical measure, perhaps a formula such as that published by Filler and Lepage, which is not dependent on anthropometric data, might be a more useful (and accurate) tool for establishing GFR in children with spina bifida. PMID- 17922292 TI - Expression levels of the mitochondrial IAP antagonists Smac/DIABLO and Omi/HtrA2 in clear-cell renal cell carcinomas and their prognostic value. AB - PURPOSE: Numerous molecular parameters are thought to be implicated in renal cell carcinoma (RCC) tumor biology and may therefore reflect the malignant potential of individual tumors. Their investigation may thus help to improve the postoperative management of RCC patients. This study characterized the mRNA expression levels and evaluated the prognostic effect of the mitochondrial inhibitor of apoptosis antagonists Smac/DIABLO and Omi/HtrA2 in tumor tissue from clear-cell RCC patients. METHODS: The relative gene expression (RGE) was analyzed by real-time RT-PCR in tumor tissue obtained from 85 patients (median follow-up: 47 months) following surgical treatment. Expression data was correlated to clinico-pathological variables and outcome. RESULTS: The RGE of Smac/DIABLO was lowest in patients with primary metastases, intermediate in those who progressed to metastatic disease, and highest in those who did not develop metastases during follow-up (P=0.006). Expression levels of Smac/DIABLO and Omi/HtrA2 were strongly correlated with each other (Pearson coefficient 0.90). Recurrence-free and tumor specific survival was shorter in patients with low Smac/DIABLO levels (P=0.019 and P=0.001) as well as in those with low Omi/HtrA2 tumor expression (P=0.033 and P=0.032). Contrary to Omi/HtrA2, low Smac/DIABLO levels were still predictive of a reduced time to recurrence (hazard rate 5.31; 95% CI: 1.16-24.21) and tumor specific survival (hazard rate 4.24; 95% CI: 1.22-14.77) in explorative multivariate analysis. CONCLUSIONS: The mRNA expression levels of the mitochondrial IAP antagonists Smac/DIABLO and Omi/HtrA2 are strongly inter correlated, but do not relate to tumor stage or grade of RCC. Our data suggest that expression of Smac/DIABLO, but not Omi/HtrA2, is inversely associated with outcome of RCC patients. PMID- 17922293 TI - Dialysate leakage into pericardium in an infant on long-term peritoneal dialysis. AB - We report on a 2-year-old boy on automated peritoneal dialysis (PD) with a history of multiple hernias and dialysate leaks who developed pericardial effusion. Magnetic resonance imaging (MRI) demonstrated a peritoneo-pericardial fistula. Dialysis had to be discontinued, since head-down tilt reproducibly induced significant hypotension. In PD patients with pericardial effusion a peritoneo-pericardial leak should be considered. PMID- 17922295 TI - Chordomas and chondrosarcomas of the skull base: comparative analysis of clinical results in 30 patients. AB - Chordomas and chondrosarcomas occur rarely in the skull base and have been often considered commonly with respect to the management. The aim of this study was to investigate the clinical results in each series of these tumors and analyze comparative outcome. Between 1991 and 2005, 30 consecutive patients with pathological diagnosis of chordoma (n=19) or chondrosarcoma (n=11) of the skull base were managed by multimodal treatment combining surgical resection with conventional photon radiotherapy and/or gamma knife radiosurgery. A retrospective analysis was conducted on these patients (aged 3 to 69 years; mean age of 37.3 years; 17 females and 13 males; mean follow-up of 56.1 months). Four deaths occurred among the patients with chordoma. The progression-free survival rate at 3 and 5 years was 61.5 and 40.0% in chordoma. In contrast, no mortalities were observed, and there was only one recurrence among the patients with chondrosarcoma. The progression-free survival rate at 3 and 5 years was 88.9 and 80.0% in chondrosarcoma. Among 15 survivors with chordoma, four patients were suffering from severe disability with progressive disease. On the other hand, most patients with chondrosarcoma harbored stable disease and less disabling symptoms except one instance of recurrence. As the biological behavior of chordoma is much more aggressive than that of chondrosarcoma in the skull base, one should make a distinction between these entities to plan an optimal treatment strategy. PMID- 17922296 TI - Sympatric diploid and hexaploid cytotypes of Senecio carniolicus (Asteraceae) in the Eastern Alps are separated along an altitudinal gradient. AB - We explored the fine-scale distribution of cytotypes of the mountain plant Senecio carniolicus along an altitudinal transect in the Eastern Alps. Cytotypes showed a statistically significant altitudinal segregation with diploids exclusively found in the upper part of the transect, whereas diploids and hexaploids co-occurred in the lower range. Analysis of accompanying plant assemblages revealed significant differences between cytotypes along the entire transect but not within the lower part only, where both cytotypes co-occur. This suggests the presence of ecological differentiation between cytotypes with the diploid possessing the broader ecological niche. No tetraploids were detected, indicating the presence of strong crossing barriers. PMID- 17922297 TI - Variation in the peacock's train shows a genetic component. AB - Female peafowl (Pavo cristatus) show a strong mating preference for males with elaborate trains. This, however, poses something of a paradox because intense directional selection should erode genetic variation in the males' trains, so that females will no longer benefit by discriminating among males on the basis of these traits. This situation is known as the 'lek paradox', and leads to the theoretical expectation of low heritability in the peacock's train. We used two independent breeding experiments, involving a total of 42 sires and 86 of their male offspring, to estimate the narrow sense heritabilities of male ornaments and other morphometric traits. Contrary to expectation, we found significant levels of heritability in a trait known to be used by females during mate choice (train length), while no significant heritabilities were evident for other, non-fitness related morphological traits (tarsus length, body weight or spur length). This study adds to the building body of evidence that high levels of additive genetic variance can exist in secondary sexual traits under directional selection, but further emphasizes the main problem of what maintains this variation. PMID- 17922298 TI - Species-level identification of Bacillus strains isolates from marine sediments by conventional biochemical, 16S rRNA gene sequencing and inter-tRNA gene sequence lengths analysis. AB - The aim of this study was to compare the ability of commonly used conventional biochemical tests, sequencing analysis of 16S rRNA genes and tDNA-intergenic spacer length polymorphism (tDNA-PCR) to identify species of the genus Bacillus recovered from marine sediments. While biochemical tests were not sufficiently sensitive to distinguish between the 23 marine strains analyzed, partial 16S rRNA gene sequences allowed a correct identification, clustering them into four species belonging to Bacillus licheniformis (n = 6), Bacillus cereus (n = 9), Bacillus subtilis (n = 7) and Bacillus pumilus (n = 1). The identification results obtained with 16S rRNA sequencing were validated by tDNA-PCR analysis of 23 marine isolates that were identified by the similarities of their fingerprints to those of reference strains. tDNA-PCR fingerprinting was as discriminatory as 16S rRNA sequencing analysis. Although it was not able to distinguish among the species of the B. cereus and B. subtilis groups, it should be considered a rapid and easy approach for the reliable identification of unknown Bacillus isolates or at least for the primary differentiation of Bacillus groups. PMID- 17922299 TI - Structure and organization of phycobilisomes on membranes of the red alga Porphyridium cruentum. AB - In the present work, electron microscopy and single particle averaging was performed to investigate the supramolecular architecture of hemiellipsoidal phycobilisomes from the unicellular red alga Porphyridium cruentum. The dimensions were measured as 60 x 41 x 34 nm (length x width x height) for randomly ordered phycobilisomes, seen under high-light conditions. The hemiellipsoidal phycobilisomes were found to have a relatively flexible conformation. In closely packed semi-crystalline arrays, observed under low-light conditions, the width is reduced to 31 or 35 nm, about twice the width of the phycobilisome of the cyanobacterium Synechocystis sp. PCC 6803. Since the latter size matches the width of dimeric PSII, we suggest that one PBS lines up with one PSII dimer in cyanobacteria. In red algae, a similar 1:1 ratio under low-light conditions may indicate that the red algal phycobilisome is enlarged by a membrane-bound peripheral antenna which is absent in cyanobacteria. PMID- 17922300 TI - Effect of quinones on formation and properties of bacteriochlorophyll c aggregates. AB - Chlorosomes of green photosynthetic bacterium Chlorobium tepidum contain aggregates of bacteriochlorophyll c (BChl c) with carotenoids and isoprenoid quinones. BChl aggregates with very similar optical properties can be prepared also in vitro either in non-polar solvents or in aqueous buffers with addition of lipids and/or carotenoids. In this work, we show that the aggregation of BChl c in aqueous buffer can be induced also by quinones (vitamin K(1 )and K(2)), provided they are non-polar due to a hydrophobic side-chain. Polar vitamin K(3, )which possess the same functional group as K(1 )and K(2), does not induce the aggregation. The results confirm a principal role of the hydrophobic interactions as a driving force for the aggregation of chlorosomal BChls. The chlorosomal quinones play an important role in a redox-dependent excitation quenching, which may protect the cells against damage under oxygenic conditions. We found that aggregates of BChl c with vitamin K(1 )and K(2) exhibit an excitation quenching as well. The amplitude of the quenching depends on quinone concentration, as determined from fluorescence measurements. No lipid is necessary to induce the quenching, which therefore originates mainly from interactions of BChl c with quinones incorporated in the aggregate structure. In contrast, only a weak quenching was observed for dimers of BChl c in buffer (either with or without vitamin K(3)) and also for BChl c aggregates prepared with a lipid (lecithin). Thus, the weak quenching seems to be a property of BChl c itself. PMID- 17922301 TI - The PucC protein of Rhodobacter capsulatus mitigates an inhibitory effect of light-harvesting 2 alpha and beta proteins on light-harvesting complex 1. AB - Rhodobacter capsulatus contains lhaA and pucC genes that have been implicated in light-harvesting complex 1 and 2 (LH1 and LH2) assembly. The proteins encoded by these genes, and homologues in other photosynthetic organisms, have been classified as the bacteriochlorophyll delivery (BCD) family of the major facilitator superfamily. A new BCD family phylogenetic tree reveals that several PucC, LhaA and Orf428-related sequences each form separate clusters, while plant and cyanobacterial homologues cluster more distantly. The PucC protein is encoded in the pucBACDE superoperon which also codes for LH2 alpha (PucA) and beta (PucB) proteins. PucC was previously shown to be necessary for formation of LH2. This article gives evidence indicating that PucC has a shepherding activity that keeps the homologous alpha and beta proteins of LH1 and LH2 apart, allowing LH1 to assemble properly. This shepherding function was indicated by a 62% reduction in LH1 levels in DeltaLHII strains carrying plasmids encoding pucBA along with a C terminally truncated pucC gene. More severe reductions in LH1 were seen when the truncated pucC gene was co-expressed in the presence of C-terminal PucC::PhoA fusion proteins. It appears that interaction between truncated PucC::PhoA fusion proteins and the truncated PucC protein disrupts LH1 assembly, pointing towards a PucC dimeric or multimeric functional unit. PMID- 17922302 TI - Atomic force microscopy reveals multiple patterns of antenna organization in purple bacteria: implications for energy transduction mechanisms and membrane modeling. AB - Recent topographs of the intracytoplasmic membrane (ICM) of purple bacteria obtained by atomic force microscopy (AFM) have provided the first surface views of the native architecture of a multicomponent biological membrane at submolecular resolution, representing an important landmark in structural biology. A variety of species-dependent, closely packed arrangements of light harvesting (LH) complexes was revealed: the most highly organized was found in Rhodobacter sphaeroides in which the peripheral LH2 antenna was seen either in large clusters or in fixed rows interspersed among ordered arrays of dimeric LH1 reaction center (RC) core complexes. A more random organization was observed in other species containing both the LH1 and LH2 complexes, as typified by Rhododspirillum photometricum with randomly packed monomeric LH1-RC core complexes intermingled with large, paracrystalline domains of LH2 antenna. Surprisingly, no structures that could be identified as the ATP synthase or cytochrome bc (1) complexes were observed, which may reflect their localization at ICM vesicle poles or in curved membrane areas, out of view from the flat regions imaged by AFM. This possible arrangement of energy transducing complexes has required a reassessment of energy tranduction mechanisms which place the cytochrome bc (1) complex in close association with the RC. Instead, more plausible proposals must account for the movement of quinone redox species over considerable membrane distances on appropriate time scales. AFM, together with atomic resolution structures are also providing the basis for molecular modeling of the ICM that is leading to an improved picture of the supramolecular organization of photosynthetic complexes, as well as the forces that drive their segregation into distinct domains. PMID- 17922303 TI - Occurrence of aflatoxin in three maize (Zea mays L.) hybrids over 5 years in Northern Mississippi. AB - Aflatoxins are produced as secondary metabolites under conducive climatic conditions by Aspergillus flavus. The incidence of aflatoxin varies with environmental conditions, genotype, and location. An expanded understanding of the interaction of the plant, fungus, and weather conditions is needed to further elucidate the field infection process of maize by A. flavus and subsequent aflatoxin contamination. One of the problems in evaluating maize hybrids for resistance to kernel infection and aflatoxin contamination is identifying a time period and environmental conditions that are most advantageous. Three maize genotypes (Pioneer Brand 3223, Mo18W x Mp313E, and Mp313E x Mp420) were evaluated from 1998 to 2002 in response to A. flavus inoculation and aflatoxin contamination and weather conditions favorable for aflatoxin contamination were identified. The highest aflatoxin levels were observed in 1998 and 2000 (1186 and 901 ng g(-1); P < 0.0001); while the lowest levels were detected in 1999 (39 ng g(-1)). Pioneer 3223 had significantly higher levels (1198 ng g(-1)) than Mp313E x Mp420 (205 ng g(-1)), and Mo18W xMp313E (161 ng g(-1); P < 0.0001). The hybrids had six weather-related variables in common that were positively correlated with aflatoxin accumulation. Four of these occurred during 65-85 days after planting and were temperature-related. These results suggest that regardless of the hybrid's maturity or physiological development, the time from 65 to 85 days after planting may be indicative of a period of stress which leads to greater aflatoxin accumulation at harvest. PMID- 17922304 TI - Respondent satisfaction regarding SF-36 and EQ-5D, and patients' perspectives concerning health outcome assessment within routine health care. AB - OBJECTIVE: To investigate respondent satisfaction regarding SF-36 and EQ-5D and patients' perspectives concerning health outcome assessment within routine health care. METHODS: Eighteen Swedish hospitals participated in the study which included 30 patient intervention groups (e.g. education groups for patients with ischemic heart disease or chronic obstructive pulmonary disease). Patients responded to SF-36 and EQ-5D before and after ordinary interventions (n = 463), and then completed an evaluation form. RESULTS: Regarding respondent satisfaction, most patients found both questionnaires easy to understand (70% vs. 75% for SF-36 and EQ-5D respectively), easy to respond to (54% vs. 60%), and that they gave the ability of describing their health in a comprehensive way (68% for both). Health outcome assessment in routine health care was perceived as valuable by 57% of the patients, while 4% disapproved. Most patients (68%) considered both questionnaires equally suitable; 25% preferred SF-36 and 8% EQ-5D. Among those who were more satisfied with a short questionnaire (EQ-5D), several still preferred a longer and more comprehensive questionnaire (SF-36). CONCLUSION: Health outcome assessment within routine health care seems to be acceptable, and even appreciated, by patients. Questionnaire length and ease of response were not found to be crucial arguments in choosing between SF-36 and EQ-5D. PMID- 17922306 TI - Cell-type specific regulation of the human 11beta-hydroxysteroid dehydrogenase type 1 promoter. AB - The intracellular enzyme 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) converts cortisone into the more active metabolite cortisol. Overexpression of 11beta-HSD1 was associated with features of the metabolic syndrome such as obesity or impaired glucose tolerance. Despite this considerable impact of 11beta HSD1, the human 11beta-HSD1 promoter has not been described in detail yet. We therefore cloned eight different promoter fragments of the 5'-upstream region of the known transcription/translation-start up to -3034 bp into the luciferase reporter vector pGL3. A low-cost in-house assay was developed and validated to detect firefly and renilla luciferase activity. Promoter fragments were analysed in human HepG2 and undifferentiated and differentiated murine 3T3-L1 cells. A differential regulation of the human 11beta-HSD1 promoter depending upon the cell type was observed. Specifically, a strong repressor of the basal promoter activity was found between -85 and -172 bp in HepG2 cells only, while an additional repressor appeared to be active between -342 and -823 bp in both, the hepatic and the adipose cell line. The presented data suggest a cell-type specific regulation of the 11beta-HSD1 promoter, which is in agreement with existing expression data from animal and human studies. The described promoter constructs will allow subsequent studies about the role of specific hormonal, metabolic and transcription factors to finally characterise the regulation of the human 11beta-HSD1-promoter in more detail. PMID- 17922307 TI - SHOX at a glance: from gene to protein. AB - The Short Stature Homeobox-containing Gene SHOX was identified as the genetic cause of the short stature phenotype in patients with Turner Syndrome and in certain patients with idiopathic short stature. Shortly after, SHOX mutations were also associated with the growth failure and skeletal deformities seen in patients with Leri - Weill dyschondrosteosis and Langer mesomelic dysplasia. Today it is estimated that SHOX mutations occur with an incidence of roughly 1:1,000 in newborns, making mutations of this gene one of the most common genetic defects leading to growth failure in humans. This review summarises the involvement of SHOX in several short stature syndromes and describes recent advances in our understanding of SHOX functions and regulation. We also discuss the current evidence in the literature that points to a role of this protein in growth and bone development. These studies have improved our knowledge of the SHOX gene and protein functions, and have given insight into the etiopathogenesis of short stature. However, the exact role of SHOX in bone development still remains elusive and poses the next major challenge for researchers in this field. PMID- 17922308 TI - Systemic infections in multiple sclerosis and experimental autoimmune encephalomyelitis. AB - Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system (CNS). It has been suggested that viral and bacterial infections contribute to the pathogenesis of MS. This review will give an overview about the influence of viral and bacterial infections on MS and experimental autoimmune encephalomyelitis (EAE). It will focus on bacterial infections and will also emphasise therapeutic consequences such as the impact of antibiotic treatment on the course of EAE. In summary, a growing body of evidence suggests that systemic infections are a risk factor for the initiation of autoimmune processes including the induction of acute events in MS. Experimental and clinical data strongly suggest early treatment of bacterial infections in MS patients to avoid aggravation and relapse. PMID- 17922309 TI - Effects of Alzheimer's amyloid-beta and tau protein on mitochondrial function -- role of glucose metabolism and insulin signalling. AB - Alzheimer's disease (AD) is the most frequent form of dementia among the elderly and is characterized by neuropathological hallmarks of extracellular amyloid-beta (Abeta) plaques and intracellular neurofibrillary tangles composed of abnormally hyperphosphorylated microtubular protein tau in the brains of AD patients. Of note, current data illustrate a complex interplay between the amyloid and tau pathology during the course of the disease. We hypothesize a direct impact of abnormally phosphorylated tau and Abeta on proteins/enzymes involved in metabolism, respiratory chain function and cellular detoxification. Probably at the level of mitochondria, both Alzheimer proteins exhibit synergistic effects finally leading to/accelerating neurodegenerative mechanisms. Moreover, accumulating evidence that mitochondria failure, reduced glucose utilization and deficient energy metabolism occur already very early in the course of the disease suggests a role of impaired insulin signalling in the pathogenesis of AD. Thus, this review addresses also the question if mitochondrial dysfunction may represent a link between diabetes and AD. PMID- 17922310 TI - Effects of insulin-like growth factor binding proteins in bone -- a matter of cell and site. AB - The actions of the insulin-like growth factor (IGF)-system are controlled by six IGF-binding proteins (IGFBPs). The IGFBPs are thought to affect local effects of IGF-I and IGF-II due to higher affinity if compared to IGF-I receptors and due to cell-type specific IGFBP expression patterns. It was found in IGFBP knockout models that the IGFBP family is functionally redundant. Thus, functional analysis of potential effects of IGFBPs is dependent on descriptive studies and models of IGFBP overexposure in vitro and in vivo. In the literature, the role of the IGFBPs for bone growth is highly controversial and, to date, no systematic look has been taken at IGFBPs resolving functional aspects of IGFBPs at levels of cell types and specific locations within bones. Since IGFBPs are thought to represent local modulators of the IGF actions and also exert IGF-independent effects, this approach is particularly reasonable on a physiological level. By sorting the huge number of in part controversial results on IGFBP effects in bone present in the literature for distinct cell types and bone sites it is possible to generate a focused, more specific and a less controversial picture of IGFBP functions in bone. PMID- 17922311 TI - Advanced glycation endproducts influence the mRNA expression of RAGE, RANKL and various osteoblastic genes in human osteoblasts. AB - Glycation reactions resulting in the generation and accumulation of advanced glycation endproducts (AGEs) are potential mechanisms by which bone protein may be altered in vivo. AGEs accumulate in the bone increasingly with age come into close contact with osteoblasts or osteoclasts. The direct effect of AGEs on bone cells has not been thoroughly investigated. This study aimed to examine whether glycated bovine serum albumin (AGE - BSA) as an AGE modulate the mRNA expression of various genes in primary human osteoblast cultures. The following parameters were included: RAGE (receptor for AGEs), alkaline phosphatase, osteocalcin, osterix and RANKL (receptor activator of nuclear factor-kappaB ligand). Primary human osteoblast cultures were obtained from bone specimens of six patients with osteoarthrosis. Human osteoblasts were treated in AGE - BSA or control-BSA (non glycated BSA) containing medium (5 mg/ml each) over a time course of seven days. After RT-PCR the mRNA expression was measured by real-time PCR. Related to control - BSA exposure, the mRNA expression of RAGE, RANKL and osterix increased during AGE - BSA treament. For alkaline phosphatase and osteocalcin a tendency of down-regulation was found. In summary, the study presents evidence that advanced glycation end products accumulated in bone alter osteoblasts by activation the AGE - RAGE pathway (RAGE mRNA up-regulation), inducing enhanced osteoclastogenesis (RANKL mRNA up-regulation) and impaired matrix mineralization (down-regulation of alkaline phosphatase and osteocalcin mRNA). Thus, AGEs may play a functional role in the development of bone diseases (e.g. osteoporosis). PMID- 17922312 TI - Immobilization staphylococcal protein a on magnetic cellulose microspheres for IgG affinity purification. AB - The main objective of this study is to develop and evaluate the immobilization of Staphylococcal Protein A on magnetic cellulose microspheres (SPA-MCMS) for immunological capture of IgG. After cloning, expression and separation, SPA was immobilized onto MCMS to prepare a magnetic affinity media subject to the purification of IgGs. The binding capacity, binding time, leakage of SPA and its reproducibility were optimized to improve the binding efficiency with an appropriate amount and recovery of IgG. Rabbit IgG was successfully purified from serum in a single-step by SPA-MCMS with an overall recovery of 73.18% and purity of 90.27%. Therefore, this study effectively illustrated the advantages of magnetic microcarriers for rapid and efficient purification of antibodies. The separation media shows a high potential for the future development of affinity isolation and immunodiagnostic application. PMID- 17922313 TI - Polyhemoglobin-fibrinogen: a novel oxygen carrier with platelet-like properties in a hemodiluted setting. AB - Polyhemoglobin (polyHb) is currently being assessed in phase III trials under various formulations. At present, none contain clotting factors or platelet substitutes to aid in hemostasis. We have prepared a novel blood substitute that is an oxygen carrier with platelet-like activity. This is formed by crosslinking fibrinogen to hemoglobin to form polyhemoglobin-fibrinogen (polyHb-Fg). This was studied and compared to polyHb for its effect on coagulation both in vitro and in vivo. In the in vitro experiments, PolyHb-Fg showed similar clotting times as whole blood, whereas polyHb showed significantly higher clotting times. This result was confirmed in in vivo experiments using an exchange transfusion rat model. Using PolyHb, exchange transfusion of 80% or more increased the normal clotting time (1-2 mins) to > 10 mins. Partial clots formed with PolyHb did not adhere to the tubing wall. With PolyHb-Fg, a normal clotting time is maintained, even with 98% exchange transfusion. PMID- 17922314 TI - Polyethylene glycol conjugated bovine hemoglobin containing 15% MetHb plays approving effect in exchange transfusion rabbit model. AB - PEG-bHb was developed by Kaizheng Biotech (Beijing, China), and pre-clinical research was completed. The objective of this study was to investigate the safe concentration of MetHb in PEG-bHb. The study was accomplished by examining the effects of PEG-bHb containing 5%, 8%, 15%, and 25% methemoglobin (MetHb), respectively, on cardiovascular system, blood chemistry, pathology of liver and kidney in rabbits following a 50% exchange transfusion. The results showed that PEG-bHb containing 5%, 8%, 15%, and 25% MetHb could keep four groups of experimental rabbits (5/5) alive until the 8th day after 50% exchange infusion as autologous whole blood did, and were superior to dextran 40 (2/5). MetHb concentration in PEG-bHb, no more than 25%, did not affect the PEG-bHb function on resuscitation of hemorrhaged rabbits by physiological measurements and blood chemistry assays. Histology study using optic and electron microscopy showed that there were slight pathological changes in hepatocytes and renal tubule epithelia in rabbits, which were infused by PEG-bHb containing 5%, 8%, and 15% MetHb. Partial organelles collapse was observed in rabbits resuscitated by PEG-bHb containing 25% MetHb. In conclusion, PEG-bHb is safe and effective when the MetHb concentration is at or below 15%. PMID- 17922315 TI - Immune effects of decreasing low-molecular weight hemoglobin components of hemoglobin-based oxygen carriers (HBOC) in a swine model of severe controlled hemorrhagic shock. AB - Hemoglobin-based oxygen carriers (HBOCs) show potential as safe, efficacious, pre hospital resuscitation fluids. The major criticism of HBOC-201 is its vasoactive property, attributed partially to low-molecular weight (low-MW) tetrameric/dimeric (TD) hemoglobin (Hb) in HBOC solution. Here we sought to determine whether resuscitation with decreasing concentrations of low-MW Hb component of HBOC affects immune responses in hemorrhagic swine. 28 anesthetized swine underwent a soft muscle crush and controlled hemorrhage of 55% blood volume, followed by resuscitation with HBOC containing 31%, 2%, or 0.4% low-MW Hb in four 10 ml/kg infusions at 20, 30, 45 and 60 minutes before hospital arrival at 75 minutes. IL-10, cell activation and adhesion markers and CD4:CD8 ratio remained unchanged in all 3 groups compared to baseline. Leukocyte apoptosis was equally elevated across all groups. Purification from 31% to 0.4% low-MW Hb in HBOC solution did not alter immune effects in a swine model of severe controlled hemorrhagic shock. PMID- 17922316 TI - Variations in dominant antigen determinants of glutaraldehyde polymerized human, bovine and porcine hemoglobin. AB - In this study, immunogenicity of human hemoglobin (hHb), bovine hemoglobin (bHb), porcine hemoglobin (pHb) and their glutaradehyde polymerized derivatives (hPolyHb, bPolyHb and bPolyHb, respectively) were compared. The nature of the dominant antigen determinants of the chemically polymerized proteins was studied. Glutaraldehyde chemical reaction enhanced the immunogenicity of the hemoglobin derivatives. In mice, the extent of the enhancement was largely comparable among hPolyHb, bPolyHb and pPolyHb. Using the methods of semi-quantitative western blotting and quantitative protein array, it was found that most of the polycloncal antibodies raised in rodents against glutaraldehyde polymerized hemoglobin derivatives of human, bovine or porcine species only weakly or did not cross-react with the hemoglobin derivatives of the other two species, indicating that hPolyHb, bPolyHb and bPolyHb vary significantly in their dominant antigen determinants, despite very high degree of identity in their primary amino acid sequences and high similarity in their three dimensional structures. PMID- 17922319 TI - The enigma of syringomyelia. PMID- 17922317 TI - Haversian remodeling in guided bone regeneration with calcium alginate film in circular bone defect model of rabbit. AB - The objective of this study is to investigate the effect of bioabsorbable Calcium alginate film in guided bone regeneration by the study of Haversian remodeling. Circular bone defects of 5 mm diameter were created in the corners of mandibles in 35 rabbits. The defects were covered with calcium alginate film (CAF) served as the experimental group, or collagen membrane (CM) as the control group, respectively. Healing condition was analyzed with gross, histological and immunohistochemical studies after 1, 2, 4, 6 and 8 weeks. The experimental group appeared more and earlier Haversian remodeling and osteoinductive factors leading to better bone regeneration. The control group showed more macrophages, less and later Haversian remodeling, absorbed slowly, while collected fewer osteoinductive factors in the early stage. Calcium alginate film, which is a relatively cheaper material, provides better effect than the collagen membrane in bone regeneration, Haversian remodeling and quantity of osteoinductive factors. PMID- 17922322 TI - Preoperative DTI and probabilistic tractography in an amputee with deep brain stimulation for lower limb stump pain. AB - This study aimed to find out whether preoperative diffusion tensor imaging (DTI) and probabilistic tractography could help with surgical planning for deep brain stimulation in the periaqueductal/periventricular grey area (PAG/PVG) in a patient with lower leg stump pain. A preoperative DTI was obtained from the patient, who then received DBS surgery in the PAG/PVG area with good pain relief. The postoperative MRI scan showing electrode placement was used to calculate four seed areas to represent the contacts on the Medtronic 3387 electrode. Probabilistic tractography was then performed from the pre-operative DTI image. Tracts were seen to connect to many areas within the pain network from the four different contacts. These initial findings suggest that preoperative DTI scanning and probabilistic tractography may be able to assist surgical planning in the future. PMID- 17922323 TI - Thalamic deep brain stimulation in the treatment of essential tremor: a long-term follow-up. AB - Deep brain stimulation (DBS) of the nucleus ventralis intermedius thalami (Vim) in the treatment of essential tremor (ET) is well documented concerning the acute effects. Reports of the long-term effects are, however, few and the aim of the present study was to analyse the long-term efficacy of this treatment. Nineteen patients operated with unilateral Vim-DBS were evaluated with the Essential Tremor Rating Scale (ETRS) before surgery, and after a mean time of 1 and 7 years after surgery. The ETRS score for tremor of the contralateral hand was reduced from 6.8 at baseline to 1.2 and 2.7, respectively, on stimulation at follow-up. For hand function (item 11 - 14) the score was reduced from 12.7 to 4.1 and 8.2, respectively. Vim-DBS is an efficient treatment for ET, also after many years of treatment. There is, however, a decreasing effect over time, most noticeable concerning tremor of action. PMID- 17922324 TI - CSF pathways: a review. AB - The treatment of patients with cerebrospinal fluid (CSF) pathway disorders (including hydrocephalus, syringomyelia, and arachnoid cysts), requires an understanding of CSF pathways and the cerebral water system. Although outcome with these disorders has improved, only limited treatment options remain available. Ongoing research has continued to improve our understanding of the cerebral water circulation and is starting to provide insight into the pathogenesis and potential treatment options of this common group of disorders. This article reviews current concepts of CSF function and pathways, following the journey of CSF from conception to absorption. PMID- 17922325 TI - Pin site metastasis of meningioma. AB - Metastasis of meningiomas due to iatrogenic implantation of tumour cells is extremely rare and only four cases have been reported to date. In this study, we report a 45-year-old female patient who presented with meningioma metastasis at the pin site of head holder applied in the original operation. PMID- 17922326 TI - Person-centredness: conceptual and historical perspectives. AB - PURPOSE: The definition and aims of rehabilitation are both topics of frequent debate. Recently several authors have suggested defining rehabilitation and its goals in terms of 'person-centredness'. However such attempts to define rehabilitation in this way have not occurred without running into their own difficulties and criticisms. Consequently, one may question whether person centredness is a good candidate to characterize and define rehabilitation. The purpose of this article is to reflect upon the historical background and conceptual underpinnings of this term and their relevance for understanding contemporary person-centred rehabilitation. METHOD: We conducted a conceptual and historical analysis of the notion of person-centredness in relation to rehabilitation. We ask first whether person-centredness has a consistent and fixed definition and meaning? Secondly, where does person-centredness come from, what is its conceptual history and does an historical approach enable us to identify a unique source for person-centredness? RESULTS: In the context of rehabilitation, we have identified four main understandings or interpretations of the term person-centredness, each of which denotes several ideas that can be, in turn, interpreted in quite different ways. Thus the concept of person-centredness in rehabilitation has multiple meanings. The conceptual history indicates that person-centredness has diverse meanings and that it has been used in a variety of contexts somewhat unrelated to disability and rehabilitation. Moreover, there does not seem to be any strict relationship between person-centredness as it is used in the context of rehabilitation and these prior uses and meanings. CONCLUSION: Person-centredness has an ancient pedigree, but its application in the field of rehabilitation raises both practical and theoretical difficulties. It may be that rehabilitation might get a better sense of what it should be and should do by focusing less on the rhetoric of person-centredness and by putting more emphasis on the investigation and operationalization of its key conceptual components. PMID- 17922327 TI - Continuity, transition and participation: preparing clients for life in the community post-stroke. AB - AIMS: To examine issues of continuity and transition facing clients as they return to life in the community following stroke and the role of rehabilitation in this process. KEY FINDINGS AND IMPLICATIONS: The sudden onset of disability following a stroke represents a major disruption to the continuity of a person's life experience. Rehabilitation has an important role in the transition from the non-disabled to the disabled state however current rehabilitation services and outcomes post-stroke focus on functional recovery rather than on a return to meaningful roles and activities and pay little attention to the transition from the non-disabled to the disabled self. Although some current rehabilitation models address the importance of involvement in a life situation, they do not adequately address issues of the role of the environment, the nature of community, the importance of meaning and choice when thinking about life situations, and change in abilities across the life course. CONCLUSIONS: Models of rehabilitation service delivery need to move to a chronic disease management model that incorporates outcomes that are meaningful to clients, and not the assumed needs or outcomes as defined by rehabilitation professionals. PMID- 17922328 TI - Politics, policy and payment--facilitators or barriers to person-centred rehabilitation? AB - AIMS: This paper explores the tensions between politics and payment in providing affordable services that satisfy the public demand for patient-centred care. KEY FINDINGS AND IMPLICATIONS: The two main approaches taken by the UK Government to curtail the spiralling costs of healthcare have been to focus development in priority areas and to cap spending through the introduction of a fixed-tariff episode-based funding system. The National Service Framework for Long Term Neurological Conditions embraces many laudable principles of person-centred management, but the 'one-size-fits all' approach to reimbursement potentially cuts right across these. A series of tools have been developed to determine complexity of rehabilitation needs that will support the development of banded tariffs. A practical approach is also offered to demonstrate the cost-efficiency of rehabilitation services for people with complex needs, and help to ensure that they are not excluded from treatment because of their higher treatment costs. CONCLUSIONS: Whilst responding to public demand for person-centred care, we must recognize the current financial pressure on healthcare systems. Clinicians will have greater credibility if they routinely collect and share outcomes that demonstrate the economic benefits of intervention, as well the impact on health, function and quality of life. PMID- 17922329 TI - Service, advocacy and adjudication: balancing the ethical challenges of multiple stakeholder agendas in the rehabilitation of chronic pain. AB - PURPOSE: To highlight potentially conflicting roles of the rehabilitation professional in the treatment of clients with persistent pain conditions. In assisting clients requiring rehabilitation, the role of the rehabilitation professional is usually fairly clear and unambiguous. If however questions about the authenticity of the client's presenting disability have been raised, then a major role conflict can arise. Many clients present with symptoms of pain and disability yet there is no objectively discernible disease, and in these cases, the authenticity of the condition may be questioned. As rehabilitation professionals we may thus find ourselves acting in different roles: (i) We might be a clinical service provider working to reduce the client's suffering, (ii) We might become the client's advocate working to protect the client in conflicts with an insurer, (iii) We might become an adjudicator working to help the insurer detect evidence of our client's fraudulent behaviour. The principal objective of this paper is to distinguish these roles, and highlight their incompatibility. It is stressed that accreditation for and competence in clinical intervention does not guarantee or legitimize competence in advocacy or adjudication. CONCLUSIONS AND RECOMMENDATIONS: The paper concludes by suggesting that the primary role of the rehabilitation professional should be the provision of clinical service and that the adoption of the role of advocate or adjudicator may cross unacceptable ethical boundaries resulting in bringing harm, intentionally, or unintentionally, to the client. PMID- 17922331 TI - Abstracts from the Innovation in Rehabilitation: Applying theory to practice conference, Rotorua, New Zealand, 16 - 17 February 2007. PMID- 17922330 TI - Why is rehabilitation not yet fully person-centred and should it be more person centred? AB - AIMS: It is a generally shared opinion that rehabilitation is not (yet) 'fully person-centred' and that it should be more. For a certain number of authors, this deficit in person-centredness has originated from the important weight of a 'medical framework' within rehabilitation. In this paper, we will discuss this criticism and its corollary: the idea that rehabilitation is bound to choose between a non-medical and a medical paradigm, since there is a fundamental contradiction between medicine and person-centredness. In the first section of the paper, we will examine the conceptual history of rehabilitation and question whether this history can really be summarized as a 'shift from a medical approach to a person-centred approach'. In the second section, we will question assumptions and suggestions that have been made to develop person-centredness in rehabilitation. In the third section, we will discuss what might be gained but also what might be lost by reinforcing person-centredness in rehabilitation. KEY FINDINGS AND IMPLICATIONS: (i) The history of rehabilitation is complex with several stages and paradigm shifts. Furthermore, these paradigms do not succeed one another but overlap. It would therefore be erroneous to reduce the history of rehabilitation to merely a shift 'from a medical approach to a person-centred approach'. (ii) Several proposals of how to make rehabilitation more person centred are found within the literature. However, none of these appears satisfactory with each leading to theoretical and practical difficulties. (iii) Although person-centredness has unquestionably contributed to the overall progress of rehabilitation, it is not certain that more person-centredness is the solution to current challenges to rehabilitation. CONCLUSION AND RECOMMENDATIONS: In some ways, the challenge rehabilitation faces is the need to transpose and adapt a notion (person-centredness) that has emerged from fields that are in fact unrelated to disability such as, for example, clinical psychology. The difficulties encountered are therefore not so much related to the particular dominance of a 'medical model' in rehabilitation than they are to the complexities of the concept of disability. We argue that one way forward might be to clarify further the respective role of the medical and non-medical aspects of rehabilitation in ways that go beyond what has been already achieved in either the ICIDH or ICF but which is still unsatisfactory or incomplete in many respects. PMID- 17922334 TI - Endothelial dysfunction in post-myocardial infarction patients with various expressions of risk factors. AB - It has not been established yet whether patients who suffer myocardial infaction (MI) in the absence of classic risk factors also have endothelial dysfunction (ED), as has been shown for patients with risk factors, and if so, to what extent it is manifested. Young male patients in the stable phase after MI were included in the study. At the time of MI, 20 patients had high and 21 patients low expression of risk factors. The control group consisted of 35 healthy age-matched males. ED was estimated by ultrasound measurement of the endothelium-dependent dilation of the brachial artery, induced by the reactive hyperemia test. Compared to the control group, the level of endothelium-dependent vasodilation was significantly reduced in both groups of patients (controls: 9.1% +/- 5.6%; patients with high risk: 5.5% +/- 5.1%; patients with low risk: 5.6 +/- 3.5 %; ANOVA, p<.01). There was no difference between both groups of patients. These results showed that ED is not associated or due only to classic risk factors. It appears that ED may occur and precede development of atherosclerosis in the absence of classic risk factors. These novel findings can have important clinical implications. PMID- 17922333 TI - Metalloproteinases in diabetics and nondiabetics during acute coronary syndromes and after 3 months. AB - The authors hypothesized that matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1, and TIMP-2 would be abnormal in acute coronary syndromes (ACS). Forty-six diabetic and 78 nondiabetic patients during ACS and after 3 months were enrolled in this study. MMP-2, -9 and TIMP-1, -2 plasma levels were measured. Significant decrease of MMP-2, TIMP-1, and TIMP-2 plasma levels was observed in the nondiabetic group with ACS after 3 months compared to the baseline value. Significant decrease of MMP-2, MMP-9, TIMP-1, and TIMP-2 plasma levels was observed in the diabetic group with ACS after 3 months compared to the baseline value. MMP-9, TIMP-1, and TIMP-2 plasma levels were higher in diabetic patients during ACS compared to nondiabetic patients during ACS. TIMP-1 and TIMP-2 increases were observed in diabetic patients with ACS at 3 months compared to nondiabetic patients after ACS. MMPs and TIMP-1 and -2 plasma levels were alterated in nondiabetic and diabetic patients during ACS and after 3 months, which may reflect abnormal extracellular matrix metabolism in diabetes during and after acute event. PMID- 17922335 TI - Vascular remodeling and prothrombotic markers in subjects affected by familial combined hyperlipidemia and/or metabolic syndrome in primary prevention for cardiovascular disease. AB - Recent evidences suggest that modulation of vascular structure by matrix metalloproteinases (MMPs) could be a main determinant of acute cardiovascular events in high-risk subjects. The authors consecutively selected 46 subjects affected by familial combined hyperlipidemia (FCH), 44 by metabolic syndrome (MS), 44 by FCH and MS, and 40 healthy subjects. All these subjects were firstly diagnosed and not treated with lipid-lowering, antihypertensive, or antidiabetic drugs. A 12-h fasting blood sample was obtained from each patient, and plasma levels of MMP-2 and MMP-9 were measured together with their tissue inhibitors and a full set of laboratory cardiovascular disease markers. MMP-2 plasma levels were not significantly different among the considered groups. MMP-9, tissue inhibitor of MMP (TIMP)-1, and TIMP-2 are significantly higher in FCH (p < .001) and MS (p < .001) patients than in healthy controls, and they are also higher in MS patients than in FCH ones (p < .001). TIMP-1 (p < .001) and TIMP-2 (p < .001), but not MMP-9, are also significantly higher in subjects with MS associated to FCH than in patients with MS alone. No specific correlation among MMPs, TIMPs, and the other studied parameters has been observed in the whole sample and in the four above-defined subgroups. MMP-9, TIMP-1, and TIMP-2 plasma levels could be significant determinant and/or diagnostic markers of MS but not of FCH. However, the superposition of MS on FCH further increases the plasma level of these parameters. The prognostic value of this observation has to be evaluated. PMID- 17922336 TI - Noninvasive assessment of endothelial activity in patients with peripheral arterial disease and cardiovascular risk factors. AB - Microvascular endothelial activity (EA) after stimulation with iontophoretically administered acetylcholine was evaluated using laser Doppler fluxmetery (LDF) and calibrated photoplethysmography (c-PPG) in normal patients and patients with peripheral artery disease (PAD). The patients included 79 non-PAD subjects and 51 patients with PAD. Upper and lower extremity EA was examined using LDF and c-PPG after acetylcholine iontophoresis for 10 min. Sensitivity and specificity were assessed using integrated area under response curve. In non-PAD patients, the EA by LDF in the upper extremity was significantly lower in the older patients compared to the younger patients. Conversely, EA by LDF detected no significant difference between these groups in the lower extremity.With c-PPG, the EA was slightly reduced in the upper but not in the lower extremity in older patients. Comparing PAD patients to the older patients, there was a significantly lower EA response in the upper and lower extremities by LDF. Likewise, c-PPG detected a highly significantly reduced EA in the upper and lower extremities for PAD patients. These results indicated that using a noninvasive technique to determine EA, there were significant differences in the EA response to acetylcholine between those with PAD and normal patients over the age of 50. Importantly, the EA response was reduced in the upper and lower extremities, indicating systemic disease of the endothelium in PAD patients. PMID- 17922337 TI - Effect of resistin on vascular endothelium secretion dysfunction in rats. AB - Resistin, a novel adipokine, was recently suggested to be involved in the development of endothelial dysfunction. However, the mechanisms of how resistin works are still unknown. This study was performed to investigate the relationship between resistin and phosphatidylinositol 3-kinase (PI3K), with the aim of gaining insight to the mechanisms by which resistin induces changes of secretion function of vascular endothelium. This study was conducted on 60 male 4-week-old Sprague-Dawley rats, which were randomly divided into four groups: resistin group (RS; n = 8), normal saline group (NS; n = 8), high-fat diet group (HF; n = 36), and control group (CO; n = 8). The resistin group was administered two injections of rat recombinant resistin. The diet-induced hyperresistinemia rats were selected from the HF group after the HF group was administered a high-fat diet for 8 weeks. The diet-induced hyperresistinemia rats were randomized into the antibody group (AB; n = 8) and hyperresistinemia group (HR; n = 8). The antibody group was given injections of resistin antibody twice per day and for 3 days. Immunohistochemistry was employed to examine the expression of PI3K p85alpha subunit and endothelial nitric oxide synthase (eNOS) in thoracic artery endothelium. In the resistin group, the levels of endothelin (ET), plasminogen activator inhibitor (PAI), and von Willebrand factor (vWF) were higher and NO was lower than those in the normal saline group. The NO level increased and ET, PAI, and vWF levels decreased in the antibody group when compared with the hyperresistinemia group. After administration of resistin antibody, the expression of PI3Kp85alpha and eNOS proteins in the antibody group was significantly increased but still differed significantly from those in the control group. PI3K grey value was correlated with resistin, PAI-1, vWF, NO, and the expression of eNOS (p < .05), after controlling for the effect of insulin. Resistin can affect the protein expression of PI3Kp85alpha, stimulate release of PAI-1, vWF, and ET, and down-regulate eNOS. The effect of resistin on PI3K signaling pathway might contribute to the development of endothelial secretion dysfunction in young rats. PMID- 17922338 TI - Endothelial connexin 37, connexin 40, and connexin 43 respond uniquely to substrate and shear stress. AB - Endothelial connexins have been linked to atherosclerosis and hypertension; however, little is know about their sensitivity to stimuli and individual functions. This study investigates the responses of endothelial connexin 37, connexin 40, and connexin 43 (Cx37, Cx40, and Cx43) to shear stress and substrate. Human endothelial cells were seeded on adsorbed collagen or a collagen gel containing smooth muscle cells and exposed to static or laminar shear stress. Connexin mRNA, protein, and gap junction communication were examined. Endothelial monolayers were treated with connexin-specific short interfering RNA (siRNA) and evaluated for communication, proliferation, and morphology under static and shear stress. Results show differential responses of Cx37, Cx40, and Cx43 to substrate and shear stress with reduced communication after shear exposure. RNA interference of individual connexins resulted in expression change of nontarget connexins, which suggests linked expression. Gap junction communication under static conditions is reduced following Cx43 siRNA treatment. Endothelial cells are more elongated with RNA interference (RNAi) targeting Cx40. In conclusion, endothelial connexins demonstrated novel sensitivity to mechanical environment and substrate. Individual isotypes show differential responses and RNAi knockdown provides new insight into connexin function and potential roles in the vasculature. PMID- 17922339 TI - Comparison of effects of epigallocatechin-3-gallate on hypoxia injury to human umbilical vein, RF/6A, and ECV304 cells induced by Na(2)S(2)O(4). AB - Hypoxia is related to the etiology of numerous pathological disease states, such as the formation of tumors or diverse retinopathies. Epigallocatechin-3-gallate (EGCG), a potent polyphenolic antioxidant and antiangiogenic compound found in green tea, has been shown to suppress the growth of blood vessels necessary for the growth of tumors and the induction of retinopathies. However, only a few studies have been carried focusing on the protective effects of EGCG on hypoxia induced injury of cultured endothelial cells. The present study investigated the effects of EGCG on Na(2)S(2)O(4)-induced hypoxic injury in three types of cultured endothelial cells, primary isolates of normal human umbilical vein endothelial cells (HUVECs), and two transformed endothelial cells lines, RF/6A and ECV304. Our results indicated that Na(2)S(2)O(4) inhibited the growth of HUVE, RF/6A, and ECV304 cells in a dose-dependent manner; EGCG also exerted inhibitory effects on the growth of the three cell types, but the toxicity of EGCG to HUVECs was less than to RF/6A and ECV304 cells. The viability of HUVE, RF/6A, and ECV304 cells treated with EGGC were the lowest at 24, 24, and 36 h, respectively, and the IC(50) of EGCG were 420 +/- 8.0, 125 +/- 7.1, and 75 +/- 5.1 microM, respectively. Furthermore, EGCG, an efficient nontoxic agent, protected all three cell types from Na(2)S(2)O(4)-induced hypoxia injury, providing partial protection from hypoxia-induced injury in normal endothelial cells at 100, 30, and 10 microM for HUVE, RF/6A, and ECV304 cells, respectively. PMID- 17922340 TI - Atorvastatin neutralizes the up-regulation of thrombospondin-1 induced by thrombin in human umbilical vein endothelial cells. AB - Statins have been reported to affect blood vessel formation. Thrombospondin-1 (TSP-1) is a multifunctional protein that affects vasculature systems such as platelet activation, angiogenesis, and wound healing. This study was designed to investigate the effect of atorvastatin on TSP-1 synthesis in thrombin-stimulated human umbilical vein endothelial cells (HUVECs), and its regulation by mevalonate or its derivatives. The results showed that atorvastatin down-regulated TSP-1 expression in HUVECs. This effect was fully reversed by mevalonate, farnesylpyrophosphate (FPP), and gerarylgeranylpyrophosphate (GGPP). Furthermore, farnesyltransferase and geranylgeranyltransferase inhibitors decreased TSP 1expression. It was also found that thrombin increased TSP-1 expression in HUVECs. Atorvastatin (0.1, 1, and 10 muM) decreased TSP-1 in thrombin-stimulated cells (45%, 66%, and 80%). Mevalonate partially reversed this inhibitory effect of atorvastatin on TSP-1, whereas the presence of FPP and GGPP did not alter TSP 1. Rho-kinase inhibitor neutralized the up-regulation of TSP-1 induced by thrombin. In conclusion, atorvastatin inhibits TSP-1 expression in endothelial cells via the mevalonate pathway. Rho protein activation is necessary for up regulation of TSP-1 synthesis induced by thrombin. Because FPP and GGPP are essential for the activity of Rho proteins, inhibition of these proteins may constitute the mechanism by which atorvastatin inhibits thrombin up-regulated TSP 1 expression. PMID- 17922341 TI - EP2306 and EP2302, two novel squalene synthase inhibitors, stimulate endothelial nitric oxide synthase expression in cultured endothelial cells. AB - EP2306 and EP2302 are two novel squalene synthase inhibitors with hypolipidemic, antiatherosclerotic, and antioxidant properties. In the present study, the authors investigated their effect on the expression and activity of endothelial nitric oxide synthase (eNOS) in cultured bovine aortic endothelial (BAE) cells and calf pulmonary artery endothelial (CPAE) cells. eNOS concentration was determined by immunoassay and eNOS activity by measuring the conversion of [(3)H]arginine to [(3)H]citrulline. Basal levels of eNOS in untreated BAE cells were 13.3 +/-1.6 ng/mg protein. Stimulation for 4 h with 30 microM of EP2306 or EP2302 resulted in increased eNOS protein level to 40% +/- 10% (p<.05) or 165% +/ 15% (p < .05) of unstimulated levels, respectively. Basal levels of eNOS in untreated CPAE cells were 3.4 +/- 0.4 ng/mg protein. Stimulation of CPAE cells for 4 h with 30 microM of EP2306 or EP2302 resulted in increased eNOS protein level to 195% +/- 24% (p < .05) and 152% +/- 19% (p < .05) of unstimulated levels, respectively. Despite their stimulatory action on eNOS expression, EP2300 compounds failed to induce any significant changes on eNOS enzymatic activity in BAE and CPAE cells. The finding that EP2300 compounds significantly increase the accumulation of eNOS in cultured endothelial cells sheds some light into their mechanism of action and supports a possible protective role of these compounds in atherosclerosis-related diseases. PMID- 17922342 TI - Cigarette smoke-induced alterations in endothelial nitric oxide synthase phosphorylation: role of protein kinase C. AB - Endothelial nitric oxide synthase (eNOS) is regulated by phosphorylation of Ser(1177) and Thr(495), which affects NO bioavailability. Cigarette smoke disturbs the eNOS-cGMP-NO pathway and causes decreased NO production. Here the authors investigated the acute effects of cigarette smoke on eNOS phosphorylation, focusing on protein kinases (PKs). Endothelial cell culture was concentration- and time-dependently treated first with cigarette smoke buffer (CSB), then with reduced glutathione (GSH) or various PK inhibitors (H-89, LY 294002, Ro-318425, and ruboxistaurin). eNOS, phospho-Ser(1177)-eNOS, phospho Thr(495)-eNOS, Akt(PKB), and phospho-Akt protein levels were determined by Western blot. CSB increased the phosphorylation of eNOS at Ser(1177) and more at Thr(495) in a concentration- and time-dependent manner (p < .01, p < .05 versus control, respectively) and resulted in the dissociation of the active dimeric form of eNOS (p < .05). GSH decreased the phosphorylation of eNOS at both sites (p < .05 versus CSB without GSH) and prevented the decrease of dimer eNOS level. CSB treatment also decreased the level of phospho-Ser(473)-Akt (p < .05 versus control). Inhibition of PKA by H-89 did not affect CSB-induced phosphorylation, whereas the PKB inhibitor LY-294002 enhanced it at Ser(1117). The PKC blockers Ro 318425 and ruboxistaurin augmented the CSB-induced phosphorylation at Ser(1177) but decreased phosphorylation at Thr(495) (p < .05 versus CSB). Cigarette smoke causes a disruption of the enzymatically active eNOS dimers and shifts the eNOS phosphorylation to an inhibitory state. Both effects might lead to reduced NO bioavailability. The shift of the eNOS phosphorylation pattern to an inhibitory state seems to be independent of the PKA and phosphoinositol 3-kinase (PI3-K)/Akt pathways, whereas PKC appears to play a key role. PMID- 17922344 TI - The use and misuse of 'America' and the JCR impact factor. PMID- 17922343 TI - Characterization of acrolein-induced protein cross-links. AB - Lipid peroxidation products contribute to protein aggregation that occurs during oxidative stress in a number of degenerative disorders. Acrolein (ACR), a highly toxic lipid peroxidation aldehyde, is a strong cross-linking agent of cellular components such as proteins. To understand the mechanisms of oxidative stress induced protein aggregation, this study characterized the ACR modification of chain B from bovine insulin by mass spectrometry. To identify the cross-linking sites, the ACR-treated peptide was digested with a protease and the resulting peptides were analysed by liquid chromatography-tandem mass spectrometry. Inter- and intra-molecular cross-linking adducts were identified between amino groups and the side chain of histidine in the peptide. These results indicated that the ACR-induced cross-links were accompanied by two reactions, namely Michael addition and Schiff base formation. In conclusion, the use of mass spectrometric techniques provided chemical evidence for protein cross-linking with ACR. PMID- 17922345 TI - Effect of low-frequency gain and venting effects on the benefit derived from directionality and noise reduction in hearing aids. AB - When the frequency range over which vent-transmitted sound dominates amplification increases, the potential benefit from directional microphones and noise reduction decreases. Fitted with clinically appropriate vent sizes, 23 aided listeners with varying low-frequency hearing thresholds evaluated six schemes comprising three levels of gain at 250 Hz (0, 6, and 12 dB) combined with two features (directional microphone and noise reduction) enabled or disabled in the field. The low-frequency gain was 0 dB for vent-dominated sound, while the higher gains were achieved by amplifier-dominated sounds. A majority of listeners preferred 0-dB gain at 250 Hz and the features enabled. While the amount of low frequency gain had no significant effect on speech recognition in noise or horizontal localization, speech recognition and front/back discrimination were significantly improved when the features were enabled, even when vent-transmitted sound dominated the low frequencies. The clinical implication is that there is no need to increase low-frequency gain to compensate for vent effects to achieve benefit from directionality and noise reduction over a wider frequency range. PMID- 17922347 TI - Repeatability of the TEN(HL) test for detecting cochlear dead regions. AB - The aim of this study was to investigate the short-term test-retest repeatability of the TEN(HL) test. Participants comprised 15 teenagers (mean age 14 years) with long-standing severe-to-profound hearing impairment, and 20 adults (mean age 74 years) with moderate-to-severe hearing impairment. Testing was repeated using the same equipment and procedures, after an interval of less than five days. The number of ears that met the criteria for diagnosing a cochlear dead region on initial test was three (13%) and nine (22.5%) for the teenagers and adults, respectively. Overall, the number of ears that changed category on retest was two (8%) and three (7.5%) for the teenagers and adults, respectively. Except at 1.5 and 4 kHz, the number of ears that changed category at individual frequencies was less than 5% for both groups of participants. TEN(HL) test repeatability was good when the results were categorized by ear or by frequency. The majority of ears that changed category on retest just met the dead-region criteria at an isolated frequency. An immediate retest is advisable in such cases. PMID- 17922346 TI - Physiology and phenomenology of tinnitus: implications for treatment. AB - We examine a contrast in understanding tinnitus and how this impacts on treatment approaches. First, a physiological account of tinnitus is described based on disinhibition and cortical remapping following injury at the receptor level, the analog for tinnitus being the 'phantom limb pain' phenomenon. Secondly, an experimental model of tinnitus is reviewed that relies on inference from conditioning animal behaviour. Arising from this, a role for conditioning in people distressed by tinnitus has been proposed, based on the unfounded premise that, for humans, tinnitus is a neutral stimulus, the distress being due to association with other stressful events. We critique this because we believe it influences approaches to tinnitus treatment. Finally, the phenomenology of tinnitus in the human case is analysed, with its nature illuminated via a series of distinctions with hearing impairment. Tinnitus can be intrinsically stressful for some people. Understanding this emphasizes the need to involve concepts and treatment in the area of clinical psychology. A flexible coalition between clinical audiologists and clinical psychologists is proposed as fruitful for tinnitus and related rehabilitation. PMID- 17922348 TI - Learning to discriminate interaural time differences at low and high frequencies. AB - This study investigated learning, in normal-hearing adults, associated with training (i.e. repeated practice) on the discrimination of ongoing interaural time difference (ITD). Specifically, the study addressed an apparent disparity in the conclusions of previous studies, which reported training-induced learning at high frequencies but not at low frequencies. Twenty normal-hearing adults were trained with either low- or high-frequency stimuli, associated with comparable asymptotic thresholds, or served as untrained controls. Overall, trained listeners learnt more than controls and over multiple sessions. The magnitudes and time-courses of learning with the low- and high-frequency stimuli were similar. While this is inconsistent with the conclusion of a previous study with low-frequency ITD, this previous conclusion may not be justified by the results reported. Generalization of learning across frequency was found, although more detailed investigations of stimulus-specific learning are warranted. Overall, the results are consistent with the notion that ongoing ITD processing is functionally uniform across frequency. These results may have implications for clinical populations, such as users of bilateral cochlear implants. PMID- 17922349 TI - A model of the occlusion effect with bone-conducted stimulation. AB - An acoustical model using simplified ear anatomy was designed to predict the ear canal sound pressure occlusion effect in humans. These predictions were compared perceptually as well as with ear-canal sound pressure occlusion effect measurements using a foam earplug with shallow insertion, a foam earplug with deep insertion into the bony part of the ear canal, and a circumaural earmuff. There was good resemblance between model predictions and ear-canal sound pressure measurements. It was also found that all occlusion positions, even deep ear-canal occlusion, produced noticeable occlusion effects. With the bone-conduction transducer at the forehead, the perceived occlusion effect was close to that obtained from ear-canal sound pressure data in the 0.3 to 2 kHz frequency range; when the stimulation was at the mastoid the difference between the perceived and measured ear-canal sound pressure occlusion effect was around 10 dB at frequencies below 1 kHz. Further, the occlusion effect was obtained in two clinical settings: with supra-aural earphones (TDH39), and insert earphones (CIR22). Although both transducers produced occlusion effects, insert earphones produced a greater effect than surpa-aural earphones at the low frequencies. PMID- 17922350 TI - The effects of training format on earplug performance. AB - This experiment investigated the effect of small-group versus individual hearing loss prevention (HLP) training on the attenuation performance of passive insert type hearing protection devices (HPDs). A subject-fit (SF) methodology, which gave naive listeners access only to the instructions printed on the HPD product label, was used to determine real-ear attenuation at threshold (REAT) at third octave noise bands between 125-8000 Hz. REAT measurements were augmented by use of the Hearing Loss Prevention Attitude-Belief (HLPAB) survey, a field-tested self-assessment tool developed by the National Institute for Occupational Safety and Health (NIOSH). Participants were randomly assigned to one of four experimental groups, consisting of 25 listeners each, in a controlled behavioral intervention trial. There were two types of HPDs (formable and premolded) and two training formats (individual and small group). A short multimedia program, including a practice session, was presented to all 100 listeners. Results showed training to have a significant effect, for both HPDs on real-ear attenuation and attitude, but, importantly, there was no difference between small-group and individual training. PMID- 17922354 TI - Peer teaching in medical education: twelve reasons to move from theory to practice. AB - OBJECTIVE: To provide an estimation of how often peer teaching is applied in medical education, based on reports in the literature and to summarize reasons that support the use of this form of teaching. METHOD: We surveyed the 2006 medical education literature and categorised reports of peer teaching according to educational distance between students teaching and students taught, group size, and level of formality of the teaching. Subsequently, we analysed the rationales for applying peer teaching. RESULTS: Most reports were published abstracts in either Medical Education's annual feature 'Really Good Stuff' or the AMEE's annual conference proceedings. We identified twelve distinct reasons to apply peer teaching, including 'alleviating faculty teaching burden', 'providing role models for junior students', 'enhancing intrinsic motivation' and 'preparing physicians for their future role as educators'. DISCUSSION: Peer teaching appears to be practiced often, but many peer teaching reports do not become full length journal articles. We conclude that specifically 'near-peer teaching' appears beneficial for student teachers and learners as well as for the organisation. The analogy of the 'journeyman', as intermediate between 'apprentice' and 'master', with both learning and teaching tasks, is a valuable but yet under-recognized source of education in the medical education continuum. PMID- 17922355 TI - Use of case-based exams as an instructional teaching tool to teach clinical reasoning. AB - BACKGROUND: It is a very well-known fact that examinations drive learning to a great extent. The examination program is actually the 'hidden curriculum' for the students. In order to improve teaching and learning one option is to strategically use of exams. AIMS: This report of the strategic use of an innovative assessment tool in clinical problem solving domain, presents the design, format, content, students' results and evaluation of one year test results of instructive case-based exams for 6th year medical students. METHOD: Using a hybrid form of the OSCE, PMP and KFE formats, we developed a case-based stationary exam. Students were treated as advanced beginners in medical career and forced to an inquiry to use their clinical knowledge in the cases. Case discussions and question-answer sessions followed the exams. Six exams were held in 2000-2001 and 382 students participated in the study. One or two problems were used for each exam and the mean duration was 27 minutes for 7-11 stations. 17-19 observers contributed to each exam. Exams were evaluated by questionnaire based feedbacks of the students and oral feedbacks of the staff members. RESULTS: The exams were well received and rated 'fair' by the students and the format was found highly 'relevant for learning' while the content was 'instructive' and 'not difficult'. The total non-satisfactory performance rate was 2.36%. Students asked to take a similar test weekly. Although it was labor intensive, staff members appreciated the collaborative working process. CONCLUSIONS: Instructive case based exams and the following case discussions seemed a high potential and motivating teaching tool in the clinical problem solving domain for 6th year students. PMID- 17922356 TI - Student teaching: views of student near-peer teachers and learners. AB - BACKGROUND: Since teaching is an important skill for future residency training and practice, training for this role should optimally be introduced as well as practiced in medical school. Studies have not fully explored the benefits and potential challenges of more senior medical students teaching more junior medical students. We define a near-peer as a trainee who is one or more years senior to another trainee on the same level of medical education training (i.e. medical students teaching other medical students, residents teaching other residents) AIM: The aim of this study was to investigate the perceptions of medical students regarding near-peer teaching and near-peer learning and to identify relevant teacher roles for near-peer teachers at two institutions from two different countries. METHODS: The authors developed two questionnaires that were filled out by a convenience sample of students who have participated in near-peer teaching (as either learner or teacher). These questionnaires were distributed at the University Medical Centre Utrecht (UMC Utrecht), the Netherlands and the Uniformed Services University (USU), USA. RESULTS: The majority of near-peer learners and near-peer teachers from both sites identified information provider, role model, and facilitator as suitable roles for near-peer teachers. Both groups agreed that planner and resource developer may be less suitable roles for near peer teachers. CONCLUSION: Information provider, role model, facilitator appear to be appropriate roles for a near-peer teacher from the perspective of near-peer learners and teachers. Given the demands of future physicians to serve as educators for both junior physicians and patients, near-peer teaching during medical school appears to be an important curricular consideration. PMID- 17922357 TI - Successful accomplishment of educational goals with clinical experience at public primary care facilities. AB - GOAL: To compare the spectrum of clinical encounters experienced by medical students at the primary level of care in six urban public health units, and to determine the extent to which these educational experiences were sufficient to meet learning objectives proposed for a teaching module. METHOD: During the 4th year of a new six- year curriculum, 113 students cared for adults, the elderly, women and children. They were supervised by faculty and trained supervisors during three 4-hours periods a week, every other week, from January to October at six primary health units. RESULTS: There were 7198 clinical encounters (2493 for adults, 2440 for women, and 2302 for children), during a total of 37 periods, averaging 1.8 cases/student per period. The top five primary diagnoses, similar at all primary health units, included: for adults--hypertension, diabetes, upper respiratory diseases, anxiety/depression, and obesity; for children--first-year follow up, upper respiratory diseases, dermatological, and infectious diseases; for women--antenatal care, vaginal discharge, cervical cancer screening, climacteric symptoms/menstrual disorders, and family planning. CONCLUSIONS: Students were exposed to and cared for the most common conditions observed at the primary level of care, with a sufficient homogeneous clinical spectrum among six primary health units, meeting essential learning objectives related to ambulatory care. PMID- 17922358 TI - Why medical students should learn how to teach. AB - BACKGROUND: We reviewed the medical-education literature in order to explore the significance and importance of teaching medical students about education principles and teaching skills. AIMS: To discuss reasons why formal initiatives aimed at improving teaching skills should be part of the training of all physicians, and how it could begin at the medical-student level. DESCRIPTION: In this article, we propose several reasons that support formal undergraduate medical training in education principles: (1) medical students are future residents and faculty members and will have teaching roles; (2) medical students may become more effective communicators as a result of such training, as teaching is an essential aspect of physician-patient interaction; and (3) medical students with a better understanding of teaching and learning principles may become better learners. We suggest that exposure to teaching principles, skills, and techniques should be done in a sequential manner during the education of a physician, starting in medical school and continuing through postgraduate education and into practice. We outline learning objectives, teaching strategies, and evaluation methods for medical-education components in an undergraduate curriculum. CONCLUSION: Medical students' informal teaching activities accompany, facilitate, and complement many important aspects of their medical education. Formally developing medical students' knowledge, skills, and attitudes in education may further stimulate these aspects. PMID- 17922359 TI - Mission Statement Day: the impact on medical students of an early exercise in professionalism. AB - BACKGROUND: The development of professional attitudes in medical students is an important aspect of medical education. AIMS: This qualitative study describes medical students' experience of Mission Statement Day. METHOD: The study was conducted using focus groups and key informant interviews. Thematic analysis identified key words, phrases, and concepts. The data was condensed into major themes and key quotes were identified to illustrate each theme. RESULTS: The process of creating a Mission Statement was more important than the Mission Statement. Three themes were identified; the central role of patients, bonding and group formation, and student ownership and valuing of the Mission Statement. Patient involvement was critical to exploring the disease and illness experience, and to stimulating discussion about compassion and professional relationships. Role modelling by faculty highlighted the value placed on this experience by the medical school. The experience was memorable, prompting the medical students to reflect on their personal values and their decision to enter medical school. CONCLUSIONS: Creation of a Mission Statement is a powerful way to introduce students to their future professional role, identify their values, and begin to develop a sense of professional identity. This memorable experience could be expanded to help students continue their professional growth. PMID- 17922360 TI - Brazilian medical students' attitudes towards patient-centered care. AB - BACKGROUND: The doctor-patient relationship is essential to medical care (Engel 1980; Balint 1984), however, medical schools focus mainly on biomedical subjects and don't give the necessary attention to communication skills, humanistic attitudes, and professional values (Haidet et al. 2001, 2002). METHODS: Attitudes of medical students towards the doctor-patient relationship have been examined and the Patient-Practitioner Orientation Scale (PPOS) has been used to measure students' and practitioners' patient-centered beliefs, first in USA (Haidet et al. 2002), and then in several other countries (Choi & Moon 2005; Tsimtsiou et al. 2005; Shankar et al. 2006). This study aimed to examine the attitudes of Brazilian medical students towards the doctor-patient relationship by using the PPOS. The scale was translated into Portuguese and was administered to approximately 800 students in their first, fifth, seventh, ninth, tenth and twelfth semesters of medical school along with a socio-demographic questionnaire. RESULTS: A total of 738 students (>90%) completed data collection. For the entire cohort, female gender (p < 0.000), later semester in medical school (p < 0.000), primary-care specialty choice (p < 0.02) and lower familial income (p < 0.03) were significantly associated with more patient-centered attitudes. Sharing sub scores, measuring beliefs about power and information between physician and patient, were significantly more patient-centered for twelfth semester male students than for first semester males (p < 0.000), but not for female students. Caring sub-scores, which measure beliefs about attending to patient's emotions and lifestyle, did not change with the years of school. In general, Caring scores were considerably higher than Sharing scores. Total PPOS scores in Brazil are comparable to those found among medical students in the United States, however they are considerably higher than PPOS total scores in Nepal, Greece, and Korea. CONCLUSIONS: Communication skills and patient care needs to be critically examined by Brazilian teachers and students aiming to change attitudes toward the doctor-patient relationship to be more patient-centered attitudes. PMID- 17922361 TI - Functional characterization of CYP2A13 polymorphisms. AB - CYP2A13 is an efficient catalyst of metabolic activation of the human carcinogens 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN). This study investigated the functional consequences of CYP2A13 polymorphisms that result in single amino acid substitutions. Five CYP2A13 variants, namely CYP2A13*2 (R257C), CYP2A13*5 (F453Y), CYP2A13*6 (R494C), CYP2A13*8 (D158E), and CYP2A13*9 (V323L), were expressed and evaluated for coumarin binding affinity, coumarin 7-hydroxylation, and -hydroxylation of (S) NNN and NNK. In addition, the 133_134 Thr deletion variant, coded for by CYP2A13*3, was expressed but was not stable to the protein purification procedure. A 30-42% decrease in coumarin 7-hydroxylation catalytic efficiency was determined for R257C and D158E. No effect on coumarin binding or (S)-NNN metabolism was observed. Three variants, R257C, D158E, and V323L, had two- to threefold decreased catalytic efficiency for NNK -hydroxylation. CYP2A13 polymorphisms resulted in modest changes in coumarin 7-hydroxylation and NNK hydroxylation activities in vitro. Although these changes are not likely to impact in vivo metabolism, these data should aid in the interpretation and design of future epidemiology studies. PMID- 17922364 TI - [Transculturality as a paradigm for intercultural approaches in psychiatry and psychotherapy]. PMID- 17922362 TI - Effect of methamphetamine on cytochrome P450 activity. AB - Amphetamine-based drugs, including methamphetamine, are some of the most widely used illegal drugs in the world. Methamphetamine is metabolized by the cytochrome P450s, the latter also being involved in the metabolism of many drugs and other xenobiotics. The effect of methamphetamine pretreatment (10 mg kg-1, intraperitoneally once daily for 6 days) on the activity of the P450 enzymes was assessed both in the rat isolated perfused liver and in vivo. The rate of 4 hydroxydiclofenac production was significantly enhanced in vivo, indicating a possible stimulatory effect on P4502C6. Similarly, the kinetics of tolbutamide and dextromethorphan in isolated perfused rat liver indicate a significant increase in both P4502C6 and the P4502D subfamily. No significant changes in midazolam kinetic in the isolated perfused rat liver were observed. The potential for methamphetamine to cause drug interactions is of clinical relevance and, therefore, it warrants further investigation. Until further drug interaction experiments are accomplished, the co-administration of drugs with methamphetamine should be conducted with caution. PMID- 17922365 TI - [Migrants from the former Soviet Union born in German families and their relatives as patients in forensic units]. AB - OBJECTIVE: Young migrants from Russia born in German families are seen as a special risk group for drug and alcohol addiction as well as for criminality. In Russia, they were marginalized as Germans, now in Germany, they are marginalized as Russians--and they repeat in Germany the internal structures of distrust against the police and other government agencies. In prisons and in forensic units, this group of young German-Russians are strongly over represented compared with to share in the general population. In prisons, they are a problem group mainly speaking Russian language, forming mafia like clans and dealing drugs--all this prevents the rehabilitation. METHODS AND RESULTS: In sharp contrast, a forensic department succeeds in integrating these people by a consequent therapeutic community approach with highly frequent group psychotherapy, intensive work, sports, autonomy in cleaning the rooms and individual teaching possibilities in German language and other specialties with a teacher working on the ward. CONCLUSION: Despite published negative experiences, it is well possible to improve the legal and social prognosis of Russian-German migrants by applying a consequent milieu- and psychotherapeutic setting utilizing actively the resources of these patients. PMID- 17922366 TI - [Integration of immigrants into the mental health care system in Germany]. AB - OBJECTIVE: Due to the effects of globalization there has to be dealt with an increased number of immigrants in the German health care system. However, esp. the mental health care system in Germany is not sufficiently prepared for this challenge. METHODS: It will be given an overview on the current situation of mental health care of immigrants with mental disorders. RESULTS: The aspect of creating specialized care units vs. opening established institutions for immigrants will be discussed. CONCLUSIONS: Examples of good clinical practice will be highlighted. Recommendations for opening the mental health care services for immigrants in Germany will be proposed. PMID- 17922367 TI - [Subjective illness beliefs of Turkish migrants with mental disorders--specific characteristics compared to german patients]. AB - OBJECTIVE: Knowledge about culture- and migration-specific characteristics of subjective illness beliefs in Turkish patients is necessary for adequate treatment. METHODS: Analysis of subjective illness beliefs in Turkish patients (F3; F4; n = 79) in comparison to matched Germans (n = 79) using a modified version of the Illness Perception Questionnaire IPQ-R. Differences were explored by t-tests and chi(2)-tests. RESULTS: Turkish patients believed significantly stronger in a chronical timeline of illness and in negative illness consequences, while German patients believed significantly stronger in treatment control and personal control. Turkish patients more often mentioned external causes of their disease compared to Germans. CONCLUSIONS: The results provide explanations of the deficient health care situation for Turkish migrants in Germany. PMID- 17922368 TI - [Relationship between mental distress and acculturation among migrants]. AB - OBJECTIVE: Several studies have described a correlation between acculturation difficulties and mental distress, even mental disorders. The stress related to the process of acculturation can lead to mental disorders such as depression by way of a change in the activity of the HPA-axis. METHODS: This correlation was examined in different samples of a total of 402 migrants. RESULTS: A significant correlation between mental distress and acculturation stress was found in all samples, yet no significant relationship between acculturation stress and length of residency in Germany. CONCLUSIONS: This non-representative, hypothesis generating study has to be replicated for samples of patients as well as other migrant groups not in treatment. A neurobiologically based specification of the definition of acculturation stress could be of relevant importance for prevention and clinical work. PMID- 17922370 TI - [Reaching migrants for preventive care: optimization of access and utilization]. AB - Migrants belong to the hard-to-reach group in health and preventive care. Essential criteria for the sustainable effectiveness of preventive and health promotion consist in the proper selection of target groups and successfully approaching them. The knowledge of possible barriers that make the access to preventive care and health promotion more difficult, e. g. low health literacy, that means the difficulties of linguistic understanding or the low acceptance regarding the provider, is necessary in order to select adequate access possibilities to the defined target groups. Up to now, for this and particularly for the ethno-specific health behaviour of migrants in Germany information hardly exist. So far, there are only a few preventive offers which are target group focussed. The use of native speaking preventive consultants is an attempt to improve the access to preventive care for migrants by low threshold come and access-structures. PMID- 17922369 TI - [Attitudes towards psychotherapy of young second-generation Turkish immigrants living in Germany]. AB - OBJECTIVE: To assess the effect of cultural identification in Turkish immigrants living in Germany on attitudes towards psychotherapy. METHODS: The approach to psychotherapy of 139 Turkish immigrants and 164 Germans was examined. RESULTS: Overall the Turkish population reported a less positive attitude towards psychotherapy in comparison to the German population. The Turkish immigrants with a less traditional cultural background reported a more positive attitude towards psychotherapy when compared to Turkish immigrants of a more Turkish identified traditional cultural background. CONCLUSION: Results suggest that culture specific factors affect attitudes towards psychotherapy in Turkish immigrants, which indicates a need for intercultural psychotherapeutic education. PMID- 17922378 TI - [COPD--noninvasive ventilation makes training effective]. PMID- 17922371 TI - [Migration as a burdening factor for schizoaffective relapse]. AB - OBJECTIVE: Migrants do not always realize the burden of migration. Earlier negative personal experiences may gain relevance later in life, even in a new cultural or life context. This case report gives an example of different relevant factors, e.g. culture and living conditions, for the course of mental illness. METHODS: We report on a female Brazilian patient, aged 37, suffering from schizoaffective psychosis with recurrent episodes. Obviously, stressful life events, the difficult relationship to her father and the expectations of her family were the trigger for psychotic episodes. RESULTS: The process and background of migration has to be considered to better understand the patient's situation and to optimize treatment. Biological, personal, cultural and life experiences are relevant aspects for relapse prevention. CONCLUSIONS: Migrants may be confronted with disadvantages resulting from cultural and communicative difficulties as an additive burden. Support from complementary health care is necessary to prevent further social isolation. PMID- 17922379 TI - [Bronchial asthma--heat treatment permits patients to breathe easier]. PMID- 17922380 TI - [Allergies--specific immunotherapy for rifampicin allergy]. PMID- 17922381 TI - [Bronchial asthma--wheezing children: who will be chronic asthmatics]. PMID- 17922382 TI - [Non-small-cell bronchial carcinoma--prognostic value of IGF protein]. PMID- 17922383 TI - [Artificial ventilation--where severely ill people learn to breathe again]. PMID- 17922384 TI - [Bronchial asthma--significant obstruction is also found in clinically stable asthma ]. PMID- 17922385 TI - [Non-small-cell bronchial carcinoma--induction--combined with radiochemotherapy- a more certain alternative]. PMID- 17922386 TI - [Cystic fibrosis--examination of the role of allergic bronchopulmonary aspergillosis]. PMID- 17922387 TI - Cytomegalovirus reactivation in "immunocompetent" patients: a call for scientific prophylaxis. PMID- 17922388 TI - Multilane highway to congenital infection. PMID- 17922389 TI - Correlates, surrogates, and vaccines. PMID- 17922390 TI - Serotype replacement in invasive pneumococcal disease: where do we go from here? PMID- 17922391 TI - Preventing the sexual transmission of HIV-1 with topical microbicides: another piece of the equation. PMID- 17922392 TI - Cellular immunity and active human cytomegalovirus infection in patients with septic shock. AB - BACKGROUND: Human cytomegalovirus (CMV) is an important opportunistic pathogen after transplantations. In the present study, monitoring of CMV in patients with septic shock was used to discover whether T helper cell type 1 (Th1) cell and natural killer (NK) cell functions interact with CMV reactivation in patients not undergoing immunosuppressive therapy. METHODS: Thirty-eight patients with septic shock were monitored, and the 23 CMV-seropositive patients were included in this prospective study. RESULTS: Seven patients (30.4%) developed an active CMV infection despite the detection of CMV-reactive Th1 cells. After active CMV infection, the frequency of CMV-reactive Th1 cells increased from a median of 0.52% to 5.04% (P=.009). Active CMV infections were terminated without antiviral therapy within 2 weeks. In parallel, the frequency of staphylococcal enterotoxin B (SEB; superantigen)-reactive Th1 cells increased from a median of 1.11% to 8.48% (P=.027). In patients without active CMV infection, the frequency of CMV reactive (median, 0.39%) and SEB-reactive (median, 1.11%) Th1 cells did not increase. Cytotoxic NK cell activity was persistently suppressed despite the presence of CD56(+)CD16(+) NK cells. Moreover, interleukin-2 application in vitro did not restore NK cell activity. CONCLUSIONS: A proinflammatory immune response may contribute to CMV reactivation in patients with septic shock. Adaptive T cell immunity, more likely than NK cell immunity, may contribute to termination of active CMV infection without antiviral therapy in these patients. PMID- 17922393 TI - Human herpesvirus (HHV)-6 and HHV-7 infections in pregnant women. AB - BACKGROUND: Both intrauterine and sexual transmission of human herpesvirus (HHV) 6 and HHV-7 have been suggested, and congenital HHV-6 infection does occur. We prospectively studied HHV-6 and HHV-7 at multiple sites in pregnant women to determine the characteristics of these viruses at repeated time points. METHODS: Peripheral blood mononuclear cells (PBMCs), cervical secretions, placenta, and cord blood were tested by nested polymerase chain reaction (PCR) and reverse transcriptase PCR for HHV-6 and HHV-7 and by quantitative PCR for HHV-6. A control group of women was also studied. RESULTS: We enrolled 104 pregnant and 31 control women. HHV-7 DNA was detected more frequently in PBMCs from pregnant women (66.9%) than HHV-6 DNA (22.2%; P<.0001), but both were found at low rates in cervical swabs (HHV-7 vs. HHV-6 DNA, 3.0% vs. 7.5%; P=.19). Pregnant women with HHV-6 DNA present in cervical swabs had a greater odds of having HHV-6 DNA present in the blood than did pregnant women with negative cervical swabs (odds ratio, 12.9; P=.0009). HHV-6 reactivation or reinfection was suggested in 17% of pregnant women. One placental sample had active HHV-6 replication. CONCLUSIONS: Detection of HHV-6 DNA in cervical secretions is associated with HHV-6 DNA in PBMC samples. Active placental infection along with congenital HHV-6 infection was identified. PMID- 17922394 TI - A framework for assessing immunological correlates of protection in vaccine trials. AB - A central goal of vaccine research is to identify a vaccine-induced immune response that predicts protection from infection or disease. The term "correlate of protection" has been used to refer to at least 3 distinct concepts, which has resulted in confusion surrounding this topic. We propose precise definitions of these different concepts of immune correlates, using the nomenclature "correlate of risk," "level 1 surrogate of protection," and "level 2 surrogate of protection." We suggest a general framework for assessing these 3 levels of immune correlates in vaccine efficacy trials. To demonstrate the proposed principles, we analyze data from a 1943 influenza vaccine field trial, supporting Weiss strain A-specific antibody titers as a level 1 surrogate of protection. Other real and simulated examples are also discussed. PMID- 17922395 TI - Cross-protection against H5N1 influenza virus infection is afforded by intranasal inoculation with seasonal trivalent inactivated influenza vaccine. AB - BACKGROUND: Avian H5N1 influenza A virus is an emerging pathogen with the potential to cause substantial human morbidity and mortality. We evaluated the ability of currently licensed seasonal influenza vaccine to confer cross protection against highly pathogenic H5N1 influenza virus in mice. METHODS: BALB/c mice were inoculated 3 times, either intranasally or subcutaneously, with the trivalent inactivated influenza vaccine licensed in Japan for the 2005-2006 season. The vaccine included A/NewCaledonia/20/99 (H1N1), A/NewYork/55/2004 (H3N2), and B/Shanghai/361/2002 viral strains and was administered together with poly(I):poly(C(12)U) (Ampligen) as an adjuvant. At 14 days after the final inoculation, the inoculated mice were challenged with either the A/HongKong/483/97, the A/Vietnam/1194/04, or the A/Indonesia/6/05 strain of H5N1 influenza virus. RESULTS: Compared with noninoculated mice, those inoculated intranasally manifested cross-reactivity of mucosal IgA and serum IgG with H5N1 virus, as well as both a reduced H5N1 virus titer in nasal-wash samples and increased survival, after challenge with H5N1 virus. Subcutaneous inoculation did not induce a cross-reactive IgA response and did not afford protection against H5N1 viral infection. CONCLUSIONS: Intranasal inoculation with annual influenza vaccine plus the Toll-like receptor-3 agonist, poly(I):poly(C(12)U), may overcome the problem of a limited supply of H5N1 virus vaccine by providing cross protective mucosal immunity against H5N1 viruses with pandemic potential. PMID- 17922396 TI - Human coronavirus infections in rural Thailand: a comprehensive study using real time reverse-transcription polymerase chain reaction assays. AB - BACKGROUND: We sought to determine whether infections with human coronaviruses (HCoVs) 229E, OC43, HKU1, and NL63 are associated with pneumonia and to define the epidemiology of HCoV infection in rural Thailand. METHODS: We developed a real-time reverse-transcription polymerase chain reaction (RT-PCR) assay panel for the recognized HCoV types and compared HCoV infections in patients hospitalized with pneumonia, outpatients with influenza-like illness, and asymptomatic control patients between September 2003 and August 2005. RESULTS: During study year 1, 43 (5.9%) of 734 patients with pneumonia had HCoV infections; 72.1% of the infections were with OC43. During study year 2, when control patients were available, 21 (1.8%) of 1156 patients with pneumonia, 12 (2.3%) of 513 outpatients, and 6 (2.1%) of 281 control patients had HCoV infections. Compared with infection in control patients, infection with any HCoV type or with all types combined was not associated with pneumonia (adjusted odds ratio for all HCoV types, 0.67 [95% confidence interval, 0.26-1.75]; P=.40). HCoV infections were detected throughout both study years; 93.6% of OC43 infections in the first year occurred from January through March. CONCLUSIONS: HCoV infections were infrequently detected in rural Thailand by use of sensitive real-time RT-PCR assays. We found no association between HCoV infection and illness. However, we noted year-to-year variation in the prevalence of HCoV strains, which likely influenced our results. PMID- 17922397 TI - Severe acute respiratory syndrome coronavirus infection in vaccinated ferrets. AB - BACKGROUND: Development of vaccines to prevent severe acute respiratory syndrome (SARS) is limited by the lack of well-characterized animal models. Previous vaccine reports have noted robust neutralizing antibody and inflammatory responses in ferrets, resulting in enhanced hepatitis. METHODS: We evaluated the humoral immune response and pathological end points in ferrets challenged with the Urbani strain of SARS-associated coronavirus (SARS-CoV) after having received formalin-inactivated whole-virus vaccine or mock vaccine. RESULTS: Humoral responses were observed in ferrets that received an inactivated virus vaccine. Histopathological findings in lungs showed that infection of ferrets produced residual lung lesions not seen in both mock and vaccinated ferrets. SARS-CoV infection demonstrated bronchial and bronchiolar hyperplasia and perivascular cuffing in ferret lung tissue, as seen previously in infected mice. No evidence of enhanced disease was observed in any of the ferrets. All of the ferrets cleared the virus by day 14, 1 week earlier if vaccinated. CONCLUSIONS: The vaccine provided mild immune protection to the ferrets after challenge; however, there was no evidence of enhanced liver or lung disease induced by the inactivated whole-virus vaccine. The ferret may provide another useful model for evaluating SARS vaccine safety and efficacy. PMID- 17922398 TI - Age-dependent differences in IgG isotype and avidity induced by measles vaccine received during the first year of life. AB - BACKGROUND: Measles remains an important cause of death worldwide, and vaccinating individuals at an earlier age could lead to better control of the disease. However, persistence of maternal antibody and young age affect the quantity of vaccine-induced neutralizing antibody and may also affect antibody quality. METHODS: Enzyme immunoassay was used to analyze measles virus-specific IgG levels, avidity maturation, and isotype changes, using serum samples from infants who received measles vaccine at 6 months of age and measles-mumps-rubella (MMR)-II at 12 months of age (n=26), measles vaccine at 9 months of age and measles-mumps-rubella (MMR)-II at 12 months of age (n=48), or only MMR-II at 12 months of age (n=27). RESULTS: The median IgG level was lower among infants with maternal antibody than among those without maternal antibody. Compared with median avidity indices for infants aged 12 months, median values were lower for 6 month-old infants with maternal antibody (P=.0001), 6-month-old infants without maternal antibody (P=.001), 9-month-old infants with maternal antibody (P=.03), and 9-month-old infants without maternal antibody (P=.006). The median IgG3 level was highest at 6 months of age. IgG1 was predominant at 12 months. Low avidity responses at 6 or 9 months of age did not hinder higher avidity responses or the switch to IgG1 after secondary vaccination. The 2-dose regimen did not augment the response, compared with the response in infants who received 1 dose at 12 months of age. CONCLUSIONS: Avidity and isotype maturation of measles vaccine induced antibody are affected by age, providing insight into the ontogeny of the immune response to measles vaccine. PMID- 17922399 TI - Incidence of pneumococcal disease due to non-pneumococcal conjugate vaccine (PCV7) serotypes in the United States during the era of widespread PCV7 vaccination, 1998-2004. AB - BACKGROUND: Widespread use of pneumococcal conjugate vaccine (PCV7) resulted in decreases in invasive disease among children and elderly persons. The benefits may be offset by increases in disease due to serotypes not included in the vaccine (hereafter, "nonvaccine serotypes"). We evaluated the effect of PCV7 on incidence of disease due to nonvaccine serotypes. METHODS: Cases of invasive disease were identified in 8 geographic areas through the Centers for Disease Control and Prevention's Active Bacterial Core surveillance. Serotyping and susceptibility testing of isolates were performed. We calculated the incidence of disease for children aged <5 years and adults aged > or =65 years. We compared rates of serotype-specific disease before and after PCV7 was licensed for use. RESULTS: The annual incidence of disease due to nonvaccine serotypes increased from an average of 16.3 cases/100,000 population during prevaccine years (1998 1999) to 19.9 cases/100,000 population in 2004 for children aged <5 years (P=.01) and from 27.0 cases/100,000 population during prevaccine years to 29.8 cases/100,000 population in 2004 for adults aged > or =65 years (P=.05). Significant increases in the incidences of disease due to serotypes 3, 15, 19A, 22F, and 33F were observed among children during this period (P<.05 for each serotype); serotype 19A has become the predominant cause of invasive disease in children. The incidence of disease due to these serotypes also increased among elderly persons. CONCLUSIONS: The incidence of pneumococcal disease caused by nonvaccine serotypes is increasing. Ongoing surveillance is needed to monitor the magnitude of disease caused by nonvaccine serotypes, to ensure that future vaccines target the appropriate serotypes. PMID- 17922400 TI - Cytokines, pregnancy, and bacterial vaginosis: comparison of levels of cervical cytokines in pregnant and nonpregnant women with bacterial vaginosis. AB - BACKGROUND: Pregnancy has been considered to be a time of relative immune compromise. Lower-genital-tract immune response appears to be influenced by pregnancy. The objective of this study was to compare, in pregnant versus nonpregnant women, endocervical proinflammatory-cytokine expression in response to bacterial vaginosis. METHODS: Endocervical levels of interleukin (IL)-1 beta , IL-6, and IL-8 in 99 pregnant and 99 nonpregnant women, all with bacterial vaginosis and without concurrent sexually transmitted infections, were assessed by ELISA. Vaginal flora was characterized on the basis of quantitative vaginal cultures. RESULTS: Women in the 2 groups differed with respect to smoking status and microbiological constituents responsible for bacterial vaginosis. When the data were stratified by these potential confounders, the levels of all 3 proinflammatory endocervical cytokines were significantly higher in pregnant women than in nonpregnant women. CONCLUSIONS: The proinflammatory cytokine milieu in the cervix is enhanced in pregnant women with bacterial vaginosis, compared with that in nonpregnant women. The notion of pregnancy as an immune-compromised state may be anatomically compartment specific. PMID- 17922401 TI - Mast cells modulate pulmonary acute inflammation and host defense in a murine model of tuberculosis. AB - BACKGROUND: Mast cells (MCs) participate in host resistance to several pathogens, but little is known about the role played by MCs in Mycobacterium tuberculosis infection. METHODS: Compound 48/80 (C48/80)-treated mice and nontreated mice were infected intratracheally with 1 x 10(5) viable M. tuberculosis bacilli (MTB; strain H37Rv). RESULTS: Infected BALB/c mice developed an acute pulmonary inflammation and had higher levels of tumor necrosis factor- alpha , interleukin 1, keratinocyte-derived chemokine, monocyte chemotactic protein-1, and macrophage inflammatory protein-2 in the lungs by day 15. In vivo degranulation of MCs by C48/80 led to a reduction in the inflammatory reaction that was associated with a marked decline in lung proinflammatory cytokine and chemokine levels. The magnitude of the cellular immune response was also partially impaired in infected mice treated with C48/80. The number of Mycobacteria bacilli recovered from the lungs of infected mice treated with C48/80 was 1 log higher than that recovered from untreated infected mice. C48/80 treatment attenuated the granulomatous inflammation in the lung parenchyma seen in untreated MTB-infected mice. CONCLUSIONS: These findings suggest that MCs participate in host defense against M. tuberculosis infection through the production and secretion of cytokines and chemokines that play a role in the recruitment and activation of inflammatory cells in this experimental model. PMID- 17922402 TI - Cross-talk between CD31 and the signaling lymphocytic activation molecule associated protein during interferon- gamma production against Mycobacterium tuberculosis. AB - Effective host defense against tuberculosis requires Th1 cytokine responses. We studied the regulation of interferon (IFN)- gamma production during tuberculosis by investigating the role of CD31, a receptor that attenuates T cell receptor signals. After antigen stimulation, CD3(+)CD31(+) blood lymphocytes decreased in healthy donors and in tuberculosis patients with robust Th1 responses to Mycobacterium tuberculosis and IFN- gamma was secreted only by CD31(-) T cells. In contrast, in patients with weak Th1 cytokine responses to M. tuberculosis, the level of CD3(+)CD31(+) lymphocytes was increased and IFN- gamma production was low. Furthermore, the inverse relationship between CD31 expression and IFN- gamma production was in contrast to signaling lymphocytic activation molecule (SLAM) expression, an IFN- gamma inducer in tuberculosis. Interestingly, CD31 bound to SLAM-associated protein (SAP), an IFN- gamma inhibitor in tuberculosis, and when CD31 and SAP were coexpressed in lymphocytes, their association inhibited the IFN gamma response to M. tuberculosis. Thus, CD31, when binding to SAP, interferes with Th1 responses, suggesting that CD31 has a key regulatory role in the signaling pathway(s) leading to the IFN- gamma response to M. tuberculosis. PMID- 17922403 TI - The association between mannan-binding lectin gene polymorphism and clinical leprosy: new insight into an old paradigm. AB - BACKGROUND: Mannan-binding lectin (MBL), a soluble protein of innate immunity, is known to play a role in pathogen recognition and clearance. For more than a decade, it has been proposed that MBL deficiency may be protective against intracellular pathogens, such as Mycobacterium leprae. METHODS: The polymorphisms at the promoter and exon 1 regions of the MBL2 gene were assessed by polymerase chain reaction and sequencing performed on 264 patients with leprosy and 214 matched healthy control subjects from southern Brazil. RESULTS. The distribution of MBL2-gene polymorphisms in patients was significantly different from that in controls, with a decreased frequency of haplotypes/genotypes associated with low expression of circulating MBL in lepromatous patients when compared with tuberculoid patients (odds ratio [OR] for haplotypes, 0.56 [95% confidence interval {CI}, 0.33-0.93] [P=.020]; OR for genotypes, 0.31 [95% CI, 0.13-0.71] [P=.004]). The LYPA haplotype was associated with susceptibility to leprosy per se (OR, 2.25 [95% CI, 1.31-3.88] [P=.003]) and to progression to the lepromatous (OR, 2.2 [95% CI, 1.21-4.05] [P=.008]) and borderline (OR, 2.98 [95% CI, 1.29 6.87] [P=.008]) forms of the disease. CONCLUSIONS: These results suggest that MBL2-gene polymorphisms play a role in susceptibility to leprosy per se and in the clinical progression of the disease. PMID- 17922404 TI - Preferential induction of transforming growth factor-beta production in gastric epithelial cells and monocytes by Helicobacter pylori soluble proteins. AB - BACKGROUND: The cytokines induced by Helicobacter pylori, as well as the intricate balance of proinflammatory and anti-inflammatory cytokines, are relevant to the outcomes of H. pylori infection. Transforming growth factor (TGF) beta and interleukin (IL)-10 are 2 vital anti-inflammatory cytokines that regulate mucosal immunity in various inflammatory and infectious diseases. METHODS: To elucidate whether host-bacteria interaction can influence TGF-beta and IL-10 production, we investigated the expression of TGF-beta and IL-10 in various mammalian cell lines preincubated with H. pylori and other enteric bacteria. RESULTS: The amount of TGF-beta protein, but not IL-10, was significantly increased after stimulation with H. pylori, but other enteric bacteria did not induce TGF- beta production. Different H. pylori strains isolated from patients with gastritis, peptic ulcer, gastric cancer and strains with cagA or vacA isogenic mutations showed similar effects on TGF-beta induction, indicating that this effect was a constitutional characteristic of H. pylori and independent of cagA and vacA status. CONCLUSION: The results imply the presence of a protein factor (termed "TGF-beta-inducing protein") that induces production of TGF-beta. In view of the multiple effects of TGF-beta , we conclude the TGF-beta-inducing protein of H. pylori might mediate the immune response and contribute to the pathogenesis of H. pylori infection. PMID- 17922405 TI - Seminal plasma reduces the effectiveness of topical polyanionic microbicides. AB - The objective of this study was to test the activity of microbicides against herpes simplex virus type 2 (HSV-2) introduced in seminal plasma. We found that seminal plasma interfered with the activity of PRO 2000 and of cellulose sulfate, increasing by 100-fold the concentration of drug required to inhibit 90% of viral plaque formation. Seminal plasma competitively inhibited binding of the microbicides to the HSV-2 envelope. Most of the interference was found in a high molecular-weight fraction; tandem mass spectrometry identified the proteins as fibronectin-1 and lactoferrin. In a murine model, the interference translated in vivo into a loss in protection. We found that 2% PRO 2000 gel protected 100% of mice challenged intravaginally with HSV-2 introduced in PBS, whereas only 55% of mice were protected if virus was introduced in seminal plasma (P=.0007, log rank test). If these findings are reflective of what occurs in humans, modifications to microbicides to ensure that they retain activity in the presence of seminal plasma are indicated. PMID- 17922406 TI - Pharmacokinetics of sulfadoxine-pyrimethamine in HIV-infected and uninfected pregnant women in Western Kenya. AB - BACKGROUND: Sulfadoxine-pyrimethamine (SP) is among the most commonly used antimalarial drugs during pregnancy, yet the pharmacokinetics of SP are unknown in pregnant women. HIV-infected (HIV(+)) women require more frequent doses of intermittent preventive therapy with SP than do HIV-uninfected (HIV(-)) women. We investigated whether this reflects their impaired immunity or an HIV-associated alteration in the disposition of SP. METHODS: Seventeen pregnant HIV(-) women and 16 pregnant HIV(+) women received a dose of 1500 mg of sulfadoxine and 75 mg of pyrimethamine. Five HIV(-) and 6 HIV(+) postpartum women returned 2-3 months after delivery for another dose. The pharmacokinetics of sulfadoxine and pyrimethamine were compared between these groups. RESULTS: HIV status did not affect the area under the curve (AUC(0-->infinity)) or the half-lives of sulfadoxine or pyrimethamine in prepartum or postpartum women, although partum status did have a significant affect on sulfadoxine pharmacokinetics. Among prepartum women, the median half-life for sulfadoxine was significantly shorter than that observed in postpartum women (148 vs 256 h; P<.001), and the median AUC(0-->infinity) was ~40% lower (22,816 vs 40,106 microg/mL/h, P<.001). HIV status and partum status did not show any significant influence on pyrimethamine pharmacokinetics. CONCLUSION: Pregnancy significantly modifies the disposition of SP, whereas HIV status has little influence on pharmacokinetic parameters in pregnant women. PMID- 17922407 TI - A checkpoint in the cell cycle progression as a therapeutic target to inhibit HIV replication. AB - Human immunodeficiency virus (HIV) expression is boosted after T lymphocyte stimulation. It is not known, however, in which phase(s) of the cell cycle HIV is maximally expressed. We demonstrate here that cell activation induces limited HIV expression and that progression to cell proliferation is required for optimal HIV replication. We also show that the G1/S cell cycle transition is a critical checkpoint in this process and that limiting progression at this step with antiproliferative drugs suppresses HIV replication. These results identify a specific phase of the cell cycle progression that is critical for HIV expression and suggest a new discrete target for anti-HIV treatment. PMID- 17922408 TI - Human lactoferrin-derived peptide's antifungal activities against disseminated Candida albicans infection. AB - BACKGROUND: Because the human lactoferrin-derived peptide, hLF(1-11), exerts potent in vitro candidacidal activity, we investigated whether it displays antifungal activity against disseminated Candida albicans infections. METHODS: Neutropenic mice were intravenously infected with C. albicans and, 24 h later, were injected with hLF(1-11); 18 h later, the number of viable yeasts in the kidneys was determined microbiologically, the size and number of infectious foci were determined histologically, and serum cytokine levels were determined by immunoassays. RESULTS: hLF(1-11) was effective (maximum reduction, 1.5 logs) against disseminated C. albicans infections, and its antifungal activity leveled off at a concentration of 0.4 ng of hLF(1-11)/kg of body weight. The antifungal activity of hLF(1-11) was increased in mice injected with interleukin (IL)-10 neutralizing antibodies, which suggests that IL-10 reduces the antifungal activity of hLF(1-11). In agreement with this result was the finding that injection of high doses of hLF(1-11) into infected mice was accompanied by increased levels of IL-10 in serum. Microscopic analysis revealed that infectious foci in kidneys of hLF(1-11)-treated mice contained mainly blastoconidia, whereas filamentous forms were abundant in untreated mice. The peptide inhibited the in vitro morphological transition of C. albicans, in a dose-dependent manner. : hLF(1-11) is effective against disseminated C. albicans infections; and its effects on C. albicans viability and virulence and on host cells may explain this antifungal activity. PMID- 17922410 TI - Reconstruction of spatially orientated myotubes in vitro using electrospun, parallel microfibre arrays. AB - The stable culture of myogenic cells and their differentiation into myotubes in vitro is often hindered by the mechanical destabilisation of the spontaneously contractile neotissue formed, resulting in the complete loss of differentiating myotubes. Electrospun, parallel aligned nylon 6/6 microfibre arrays were use successfully for the culture of C2C12 myoblasts and their differentiation to form mechanically stable, orientated myotubes in vitro. Myoblasts adhered strongly to the parallel fibre array, forming a compact cell sheath across the entire array, aligning individual cells in parallel to the direction of the fibrous substratum. The myogenic potential of C2C12 myoblasts was not impaired and resulted in the formation of elongated myotubes expressing alpha-actinin, adult myosin heavy chain and nicotinic acetylcholine receptors as muscle-specific marker proteins. Newly formed C2C12 myotubes were themselves orientated in parallel to the direction of the underlying fibrous substratum and exhibited a high level of structural integration with the surrounding cells. In contrast, non-woven, non orientated nylon 6/6 meshes, produced by conventional electrospinning, exhibited greatly reduced levels of C2C12 myoblast attachment and adherent myoblasts did not differentiate into myotubes. In conclusion, parallel microfibre arrays provided a superior microscale topography for the stable maintenance and differentiation of myotubes in vitro. PMID- 17922409 TI - Expression of antimicrobial peptides in the normal and involved skin of patients with infective cellulitis. AB - BACKGROUND: Endogenous antimicrobial peptides participate in the innate defense of skin against a variety of pathogens. The systemic expression of these peptides in normal-appearing skin of patients with infective cellulitis is unknown. METHODS: Study patients were adults with infective cellulitis and signs of systemic inflammation. Skin biopsy and serum specimens were obtained from patients and from control subjects with no active infection. Cathelicidin and human beta-defensin 2 mRNA expression were determined by real-time polymerase chain reaction. RESULTS: Skin biopsy specimens from 11 patients and 4 uninfected control subjects were analyzed. The relative expression level for cathelicidin mRNA was elevated in both the involved and the distal normal-appearing skin of patients with cellulitis, compared with expression in the skin of control subjects (mean ratios, 39.46 vs. 1.32, P=.0059; and 21.41 vs. 1.32, P=.0059). Similarly, the relative expression level of human beta -defensin 2 mRNA was elevated in both the involved skin (mean ratios, 20,844 vs. 11.65; P=.0015) and in distal normal-appearing skin of patients with cellulitis (mean ratios, 201.1 vs. 11.65; P=.0103). DISCUSSION: In response to cutaneous infection there is a local and distal increase in endogenous antimicrobial peptide mRNA in both involved and normal-appearing skin. These observations show, for the first time to our knowledge, that after infection the human body responds by increasing systemic innate immunity. PMID- 17922411 TI - [Association between mRNA level of Pde4d and Alox5ap and hypertensive stroke as well as hypertension in rats]. AB - OBJECTIVE: To evaluate whether mRNA levels of Pde4d and Alox5ap were associated with hypertensive stroke and hypertension in stroke-prone renovascular hypertensive rats (RHRSP) which could simulate human being's hypertensive cerebral stroke. METHODS: Five groups were established: normotensive group, gradient hypertensive groups I, II and III(with contractive pressure of 140-159 mmHg, 160-179 mmHg and 180-199 mmHg respectively) and spontaneous stroke group. RNA from leukocytes in peripheral blood of each rat underwent real time PCR after reversed. RESULTS: The mRNA levels of Pde4d and Alox5ap of spontaneous stroke group were statistically higher than that of the other groups. Expression of Pde4d of hypertensive group I was a bit higher than that of normotensive group and hypertensive groups II and III; as for Alox5ap, there was no statistical difference between normotensive group and all gradient hypertensive groups. CONCLUSION: Animal experiments come to conclusions that over-expression of Pde4d and Alox5ap are associated with hypertensive stroke but not with hypertension. Therefore, the two genes confer the risk of hypertensive stroke independent of traditional risk factors. It is speculated that over-expression of Pde4d and Alox5ap can motivate onset of hypertensive cerebral stroke by participating in inflammation of arterial walls. PMID- 17922412 TI - [An analysis method to apply linkage disequilibrium maps to association study]. AB - OBJECTIVE: To apply linkage disequilibrium (LD) maps to associations studies with high throughput single nucleotide polymorphisms (SNPs). METHODS: Seven hundred and fifty-four SNPs were genotyped in 160 Shanghai Chinese. LD maps were constructed in cases and controls separately. By comparing the decline of LD unit with distance between the two groups, disease susceptible loci were estimated. This method was compared with traditional analyses including LD analysis, single SNP and haplotype analyses. RESULTS: The analysis of LD maps could detect the chromosome regions with different LD patterns between the cases and controls. The alleles and/or haplotypes frequencies of SNPs within the regions had significantly different distributions or trends of significantly different distributions. CONCLUSION: This method may be applied to analyze the data from association studies with high throughput SNPs genotype information. PMID- 17922413 TI - [BRCA1 germ line mutations in Chinese early-onset breast cancer patients]. AB - OBJECTIVE: To investigate the role of disease associated germ line mutations in BRCA1 gene among Chinese early-onset breast cancer patients. METHODS: A total of 188 early-onset breast cancer patients, who were diagnosed with breast cancer before 41-year-old, were enrolled from four breast cancer clinical centers in China. Thirty-nine of them (20.7%) also had family history of breast/ovarian cancer. DNA extracted from lymphocytes was amplified by polymerase chain reaction (PCR) for the entire exons and the splicing sites of BRCA1. Twenty-two of the patients were screened by single strand conformation polymorphism (SSCP), and the other 166 of them were screened by denaturing high performance liquid chromatography (DHPLC). The abnormal fragments recognized were ascertained by DNA direct sequencing. For those samples with the same recurrent mutation, five BRCA1 linked markers (D17S855, D17S1322, D17S1323, D17S1326 and D17S1327) were used for the allelotype analysis. RESULTS: Twelve disease-associated mutations were identified in 15 (8.0%) patients, among which BRCA1 1100delAT and 5589del8 were identified in 3 and 2 patients respectively. Nine (23.1%) of them were identified in those with breast/ovarian cancer family history. The difference of BRCA1 mutation frequency between the patients with and without family history was statistically significant (P=0.001). Allelotype analysis showed the two BRCA1 5589del8 mutation carriers shared the same allelotype in all the 5 STR sites, and two of the three 1100delAT mutation carriers, who came from the northern China, also shared the same allelotype in all the 5 STR sites, which were different from those of the 5589del8 mutation carriers'. CONCLUSION: This is a relatively very large scale multi-hospital-based study of BRCA1 mutations in Chinese early-onset breast cancer patients up to now. It seems reasonable to give genetic consultations and genetic test of BRCA1 gene to early-onset breast cancer patients in China, especially for those with breast/ovarian cancer family history. The two recurrent mutations might be founder mutations of Chinese population. It might be cost-effective to analyze these two mutations before whole gene analysis. PMID- 17922414 TI - Analysis of SOX4 gene mutation in non-small cell lung cancer tissues. AB - OBJECTIVE: To investigate the mutation of SOX4 gene in the different tumor tissues with pathological stages and types of non-small cell lung cancer (NSCLC), and to explore its roles in the progression of lung carcinoma. METHODS: The SOX4 gene HMG-box of lung cancer tissues and paracancerous tissues were amplified by PCR, 20 cases shown difference by single strand conformation polymorphyism analysis were sequenced. The DNA sequences were compared with normal sequences by software Clustal and DNAStar. RESULTS: In the 90 NSCLCs, 18 cases were found with mutations of SOX4 gene and were sequenced, and there were 2 mutational points. Seven were detected from squamous cell carcinoma, five from adenocarcinoma and six from adeno-squamous. Three were obtained from tissues in stage I, five in stage II, six in stage III, and four in stage IV. The mutation rate in stage II, III and IV was significantly higher than that in stage I. CONCLUSION: SOX4 gene mutation is not associated with pathology histological types of tumor, but it is significantly associated with pathological stages and the mutation rate increases gradually, which has relation with advanced pathological stages in NSCLC. The results indicate that the SOX4 gene mutations might be related in the lung carcinogenesis and tumor metastasis. The study also provides molecular data for study the links between the mutation of SOX gene and human oncogenesis. PMID- 17922415 TI - [Sequence analysis of a novel HLA allele B*5618]. AB - OBJECTIVE: To identify HLA novel allele in Chinese Han individual. METHODS: An unknown HLA-B allele which was similar to HLA-B*5610 was detected by polymerase chain reaction-sequence specific oligonucleotide probes(PCR-SSOP), PCR-sequence specific primer(PCR-SSP) and heterozygous sequence-based typing (SBT) in a Chinese Han individual. Its anomalous patterns suggested the possible presence of new allele. The HLA-B*56 allele was amplified separately by using allele-specific primers and sequencing exons 2-4 in both directions. The differences between the novel B*56 allele and B 5610 were identified. RESULTS: There were 4nt changes from B*5610 in exon 3, at nt379 where C>G (codon 127 CTG>GTG, 127 Leu>Val); nt412 where A>G (codon 138 AAC>GAC, 138 Asn>Asp), nt419 where T>C and nt420 where A>C (codon 140 TTA>TCC, 140 Leu>Ser). The sequence was submitted to Genbank and the accession number was EF016753. CONCLUSION: This allele is a novel HLA-B allele, and has been officially named HLA-B*5618 by the WHO Nomenclature Committee in September 2006. PMID- 17922416 TI - [Triple-color primed in situ labeling protocol of human metaphase chromosomes]. AB - OBJECTIVE: To study the feasibility of simultaneous detection for several chromosomes with optimized triple-color primed in situ labelling (PRINS) protocol in cultured peripheral blood lymphocytes. METHODS: Pre-test of gonosome detection with dual-color PRINS protocol was performed to explore and optimize the order and condition of PRINS primers. A peripheral blood sample from a Klinefelter's syndrome patient (47, XXY) had also been studied with optimized triple-color PRINS to prove the correspondence between the number of signals and chromosomes. RESULTS: Chromosome 18, X and Y had been simultaneously and specifically marked within 3 hours. The frequency of successful labeling reached 90% both in dual color and triple-color test. Two chromosome X had been correctly showed in lymphocyte sample of Klinerfelter's syndrome. CONCLUSION: Numerical chromosome anomalies could be rapidly and exactly detected with this non-ddNTP-blocking multicolor PRINS protocol in peripheral blood lymphocytes. The results of in situ labeling are much clearer with inner control. PMID- 17922417 TI - [Identification of a novel HLA allele B 4446 by sequencing-based typing]. AB - OBJECTIVE: To identify a novel HLA allele in Chinese population. METHODS: A new HLA-B allele was initially detected by an usual PCR-SSP and PCR-SSOP in routine typing HLA allele. Sequence-based typing (SBT) was used to identify and analysis the difference between the new allele and HLA-B 4409 allele. RESULTS: The HLA-B exon 3 nucleotide sequence of the novel allele was different from all other known alleles. The allele had 3 nucleotides replaced of the closest matching B 4409 allele at nt538(G>C), nt539(A>T) and nt540 (C>G) in exon 3, resulting in an amino acid change from D(GAC) to L(CTG) at codon 180. CONCLUSION: A novel HLA allele was confirmed by the SBT and it was officially designated as HLA-B 4446 by WHO Nomenclature Committee for Factors of the HLA System in September,2005. PMID- 17922418 TI - [Molecular genetic analysis of FUT1 and FUT2 gene in para-Bombay Chinese: a novel FUT1 allele is identified]. AB - OBJECTIVE: Molecular genetic analysis of FUT1 and FUT2 gene was performed for seven Chinese Han individuals serologically typed as para-Bombay. METHODS: Seven DNA samples were studied by polymerase chain reaction and then by direct sequencing. Molecular cloning sequencing was done for an individual with a novel FUT1 allele. Family segregation analysis of the novel FUT1 allele was done to explore whether the allele was responsible for the fucosyltransferase defects of H. RESULTS: The FUT1 genotypes of seven para-Bombay individuals were h1h1 (four individuals), h2h2 (two individuals), h328hnew (one individual), alleles h1 lost one of the three AG repeats located at the nucleotides 547-552 of the FUT1 gene, h2 lost two of the three T repeats located at the nucleotides 880-882, h328 (nt328G>A) was a missense mutation, all of them were known mutations, while allele hnew deleted GGTATTCCGCATCACCCTGCCCGTGCTGGCCCC at nt360-400, total 33 bases, and the frame-shift mutation was not previously reported. The segregation of the hnew allele in his family showed that his father genotype was Hh328, and his mother was Hhnew, while two brother were h328hnew. The FUT2 genotypes of seven para-Bombay individuals were Se357 Se357 (three individuals), Se357 Se357,385 (three individuals), Se357,716Se357,716(one individual), the functional Se357(nt357C>T), Se716(nt716G>A) and the weakly functional Se385(nt385A>T) were known. The seven para-Bombay individuals carried at least one copy of a functional FUT2 allele was consistent with their secretor status. CONCLUSION: A novel FUT1 allele was identified in a para-Bombay Chinese individual, which was responsible for the inactivation of the FUT1-encoded enzyme activity. PMID- 17922419 TI - [The relationship between the exon 10 of the p47(phox) polymorphism and stroke and the effect of the polymorphism on plasma lipid]. AB - OBJECTIVE: To investigate the relationship between C923T (Ala308Val) polymorphism in exon 10 of protein 47000 phagocyte oxidase(p47(phox)) gene and stroke and to evaluate the effect of C923T (Ala308Val) polymorphism on plasma lipid levels in Hunan Hans population. METHODS: Hunan Han population C923T (Ala308Val) polymorphism in p47phox gene was determined by PCR-single strand conformation polymorphism analysis and DNA sequencing in 100 healthy controls, 220 patients with stroke, and 10 stroke pedigrees. Plasma lipid levels were measured by routine methods. RESULTS: No statistically significant differences were found in frequencies of genotypes and alleles of C923T (Ala308Val) polymorphism between the controls and stroke patients (P > 0.05). The serum levels of triglyceride in cerebral infarction patients and controls with CT genotype were markedly higher than those with CC genotype (P < 0.05). CONCLUSION: There is no association between C923T (Ala308Val) polymorphism and stroke in Hunan Hans; C923T (Ala308Val) polymorphism is associated with plasma lipid metabolism in Hunan Han population with cerebral infarction. PMID- 17922420 TI - [Study on gene control region of mitochondrial DNA in familial breast cancer]. AB - OBJECTIVE: To study on mutations in D-loop region which is gene control region of mitochondrial genome in patients with familial breast cancer. METHODS: Twenty three breast cancer patients came from twenty-one families of breast cancer, and eighteen healthy controls participated in the study. PCR amplification of D-loop region in mitochondrial DNA was performed and then the product was sequenced to analyze mutations. RESULTS: One hundred and twenty-six mutations in D-loop region were found in twenty-three patients with familial breast cancer, and four mutations were new. In all of twenty-three patients, thirty-seven mutations were found in D310 which was hot spot of D-loop region in mitochondrial DNA. In these mutations, T>C in 310, TC insert in 311-312, CA deletion in 522-523 and C>G in 527 were multi-presentation mutations in patients with familial breast cancer. Mutations had no difference in the same family member of breast cancer family except that occurrence in the region of D310. In the same family, mutations in D310 of patients were different from controls. CONCLUSION: Mutations in D310 of familial breast cancer patients may enhance their susceptibility to breast cancer. PMID- 17922421 TI - [The relationship between polymorphism of gene involved in folate metabolism, homocysteine level and risk of Down syndrome]. AB - OBJECTIVE: To investigate whether the polymorphism in methylenetetrahydrofolate reductase (MTHFR) gene involved in folate metabolism is associated with Down syndrome (DS). METHODS: One hundred Chinese mothers who gave birth to babies with DS and 100 control mothers were chosen. Genotype of MTHFR 677 was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and concentration of plasma homocysteine (HCY) was measured by chemiluminescence. RESULTS: The MTHFR 677T allele frequency was significantly different among case mothers, compared with control mothers (P=0.002); the odds ratio for the heterozygous CT genotype was 2.12 (95%CI: 1.14-3.94), whereas for the homozygous TT genotype, the odds ratio was 3.43 (95%CI:1.41-8.36). The mean plasma HCY concentration [(9.04 +/- 3.85) mu mol/L] of cases was significantly different from that of controls [(6.53 +/- 2.06) mu mol/L](P <0.01). The presence of the 677C>T substitution in one or both alleles was associated with increased plasma HCY both in case mothers and control mothers (P < 0.01). Interestingly, although both being MTHFR 677CC, the plasma HCY concentrations were higher in case mothers than in control mothers, the increase was not dependent on MTHFR genotype (P < 0.01). CONCLUSION: Our results provide evidences that plasma HCY and genetic polymorphism in gene of folate pathway are risk factors for mothers to have a DS child in China. PMID- 17922422 TI - [The association of XRCC2 gene polymorphism with susceptibility to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma in China]. AB - OBJECTIVE: To investigate the possible association of single nucleotide polymorphism (SNP) at the 41657C/T position and 4234G/C position of X-ray repair cross-complementing gene 2 (XRCC2) with susceptibility to esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA) in a population of high incidence region, Ci county and She county of Hebei. METHODS: The genotypes of XRCC2 41657C/T and 4234G/C SNPs were detected by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis in 330 ESCC patients, 254 GCA patients and 629 healthy controls. RESULTS: The genotype frequency of XRCC2 41657C/T in ESCC patients (67.8%, 26.4% and 5.8%) was significantly different from that in controls (68.8%, 28.8% and 2.4%; chi square was 7.43, P was 0.02). Compared with CC genotype, TT genotype significantly increased the risk of developing ESCC (OR=2.12, 95%CI: 1.03-4.35). The genotype (59.9%, 35.8% and 4.3%) and allelotype distributions ofXRCC2 41657C/T in GCA patients were significantly different from that in controls (chi square was 7.46 and 7.23, P was 0.02 and 0.01). Compared with CC genotype, CT genotype significantly increased the risk of developing GCA (OR=1.38, 95%CI: 1.01-1.89). The genotype and allelotype distributions of the 4234G/C SNPs in ESCC and GCA patients were not significantly different from that in controls (all P values were above 0.05). Compared with GG genotype, the CG and CC genotype of XRCC2 4234G/C did not show significant effect on the risk of developing ESCC and GCA. When the two XRCC2 SNPs were combined analyzed, the haplotype distribution in GCA patients was significantly different from that in controls (chi square was 13.28, P was less than 0.01). Compared with 41657C/4234G haplotype, 41657C/4234C and 41657T/4234G haplotypes significantly increased the risk of developing GCA (OR were 1.44 and 1.55, 95%CI were 1.06-1.95 and 1.18-2.02, respectively). CONCLUSION: In high incidence region of Hebei province, we conclude that XRCC2 41657C/T polymorphism has a potential to be a susceptibility factor for ESCC and GCA while XRCC2 4234G/C polymorphism may not provide a useful marker to predict susceptibility to ESCC and GCA. However, the 41657C/4234C and 41657T/4234G haplotypes might increase the risk of developing GCA. PMID- 17922424 TI - [The population genetics study in HLA field]. AB - Human leukocyte antigen (HLA) is the most polymorphic genetic system found in human genome. The polymorphisms of different HLA genes and haplotypes in different ethnic and geographic populations are of high importance for investigation of their population genetic characteristics and searching for HLA matched unrelated hematopoietic stem cell transplantation donors, as well as in disease association studies. The HLA molecular genetic principals and the progress of HLA population investigation were reviewed, as well as the methods applied in the field. PMID- 17922423 TI - [Study on genetic susceptibility of the single nucleotide polymorphism of FCGR3A gene and systemic lupus erythematosus]. AB - OBJECTIVE: To investigate the role of FCGR3A gene in susceptibility to systemic lupus erythematosus (SLE) using family based studies. METHODS: A total of 119 patients from 95 nuclear families, with SLE according to the American College of Rheumatology 1997 criteria were recruited. In addition, 316 family members of these patients were also genotyped. A family-based association study was used to explore the association between gene polymorphism and SLE. The authors studied the single nucleotide polymorphisms (SNP) encoding non-synonymous substitution in the cFCGR3A gene with respect to genetic susceptibility to SLE. The FCGR3A gene was genotyped with RFLP. RESULTS: Among 119 SLE patients, the frequency of FCGR3A 72R/S, R and S allele were 39.4% and 60.6%; the frequency of FCGR3A R/S RR, RS and SS genotypes were 9.1%, 60.6% and 30.3%, respectively. Univariate (single marker) family-based association tests (FBATs) demonstrated that variant allele at the SNP(rs403016) in exon 3 of FCGR3A gene was significantly associated with genetic susceptibility to SLE in Additive Model(Z=2.544, P =0.01097) and Recessive Model(Z = 2.198, P = 0.02795). TDT analysis showed an excess of the allele of R from heterozygous parents to affected offspring (chi square was 9.30, P=0.0032). CONCLUSION: The findings suggest that the FCGR3A gene may be the susceptible gene of SLE in Chinese population, and that the individual carrying FCGR3A 72R allele was significantly associated with increase of susceptibility to SLE. PMID- 17922425 TI - [Another grey zone for clinical genetics: chromosomal microduplication 22q11.2]. AB - Theoretically, microduplication of chromosomal region 22q11.2, which is rich in segmental duplications, should be as frequent as microdeletions of the same region. Preliminary analysis on the rarity of reports for 22q11.2 microduplication in the literature has suggested that, for the discovery of 22q11.2 microduplication, there has been a lack of sensitivity for routine diagnostic techniques such as karyotyping, PCR and FISH. On the other hand, the diverse anomalies and extremely variable phenotypes of carriers also implied great difficulties one has to face upon clinical consultation. Genetics as well as clinical problems in connection with 22q11.2 microduplication has vividly illustrated the great challenge for the interpretation of genotype-phenotype correlation, and thereby posed yet another gray zone for clinical genetics research. PMID- 17922426 TI - [Leber's hereditary optic neuropathy is associated with the mitochondrial G11696A mutation in two Chinese families]. AB - OBJECTIVE: To report the clinical, genetic, and molecular characterization of two Chinese families with Leber's hereditary optic neuropathy (LHON). METHODS: Ophthalmological examinations showed that only probands in two families exhibited visual loss at the age of 10 and 17 years respectively. The entire mitochondrial genome of two probands was PCR amplified in 24 overlapping fragments using sets of oligonucleotide primers. RESULTS: Mutational analysis of mitochondrial DNA (mtDNA) in these pedigrees revealed the absence of three common LHON associated G11778A, G3460A and T144484 mutations but the presence of homoplastic LHON associated ND4 G11696A mutation, which was present in one out of 167 Chinese healthy controls. CONCLUSION: Sequence analysis of the complete mitochondrial genomes in two pedigrees showed the distinct sets of mtDNA polymorphisms, belonging to Eastern Asian haplogroup D4. The incomplete penetrance of visual loss and the presence of one in 167 controls suggested that this mutation itself is insufficient to produce a clinical phenotype and other modifier factors play a role in the phenotypic manifestation. The lack of functional mtDNA variants in these pedigrees ruled out the role of mitochondrial background in the phenotypic expression of visual loss. Therefore, nuclear modifier gene(s) or environmental factor(s) may play a role in the phenotypic expression of the LHON-associated G11696A mutation in two Chinese pedigrees. PMID- 17922427 TI - [Comparative study on clinical features between TEL-AML1 positive and negative childhood acute lymphoblastic leukemia]. AB - OBJECTIVE: To determine the incidence of TEL-AML1 fusion gene in childhood acute lymphoblastic leukemia (ALL) and to compare the clinical features between TEL AML1 positive and negative patients. METHODS: Samples of bone marrow or peripheral blood were collected from 95 newly diagnosed ALL children and the TEL AML1 fusion gene was detected using nested reverse transcription-polymerase chain reaction (RT-PCR). The ALL patients were stratified into TEL-AML1 positive and negative groups and the clinical features were compared. RESULTS: Among 95 patients, 20 (21.05%) were TEL-AML1 positive. The median age of TEL-AML1 positive patients was 5.9 years old and M/F ratio was 1.22:1. There were significant differences between TEL-AML1 positive and negative patients in hepatomegaly (2.75 cm vs. 4 cm below costal arch, P=0.006), splenomegaly (0 cm vs. 3 cm below costal arch, P < 0.001), initial white blood cell count (median 7.40 x 10(9)/L vs.18.70 x 10(9)/L, P=0.011), initial peripheral blood blast (median 2.45 x 10(9)/L vs.11.66 x 10(9)/L, P=0.013), hemoglobin level [(61.45 +/- 13.46) g/L vs. (75.89 +/- 23.11) g/L, P=0.003] and serum lactate dehydrogenase [(621.47 +/- 335.85) U/L vs.(1566.64 +/- 1720.45) U/L, P=0.020], while no differences were found between two groups in age, gender ratio, initial platelet count, percentage of blast in bone marrow, immunophenotypes and the expression of myeloid antigen CD13, CD33 and CD34. The prednisone sensitivity test showed that all 12 TEL-AML1 positive patients were good responders, while there were 11 prednisone poor responders among 40 negative patients (27.50%, P < 0.05). Bone marrow examination on day 15 showed no difference in the rate of complete remission between TEL-AML1 positive and negative patients. CONCLUSION: The incidence of TEL-AML1 fusion gene in cases of ALL is 21.05%. The load of leukemia cells in TEL-AML1 positive patients is significantly smaller than its counterparts, and the blast cells in TEL-AML1 positive patients are more sensitive to prednisone, indicating childhood ALL with TEL-AML1 fusion gene has a favorable prognosis. PMID- 17922428 TI - [Screening for the microdeletions of azoospermia factor on the Y chromosome in male infertile patients from Guangzhou]. AB - OBJECTIVE: To investigate the relationship between microdeletion of azoospermia factor (AZF) and male infertility. METHODS: Multiplex PCR was used to detect Y chromosome microdeletion in AZFa, AZFb and AZFc in 103 cases of idiopathic azoospermia, 72 cases of severe idiopathic oligozoospermia, and 60 healthy male controls. RESULTS: No microdeletion was found in 60 controls. Y chromosome microdeletion was found in 19 of 175 azoospermia patients, the total prevalence rate of microdeletion was 10.9%. There were 15 cases (11 for azoospermia, 4 for severe oligozoospermia) in AZFc (8.6%), 3 cases (1 for azoospermia, 2 for severe oligozoospermia) in AZFb+c (1.7%), 1 case (azoospermia) in AZFa+b+c (0.6%). According to statistics, the difference of microdeletion rate between two groups was significant(P < 0.01). CONCLUSION: Y chromosome microdeletions is an important reason of azoospermia. Screening of Y chromosome microdeletions for azoospermia patients before intracytoplasmic sperm injection treatment is essential. PMID- 17922429 TI - [Study on correlation between HLA-A, B, DR alleles and Duchenne muscular dystrophy]. AB - OBJECTIVE: To analyze the polymorphism of HLA-A, B and DR alleles of Duchenne muscular dystrophy (DMD) patients in Han nationality of South China and to discuss the role of immune and genetic factors in the pathogenesis of DMD and muscular fiber necrosis. METHODS: Polymerase chain reaction-reverse sequence specific oligonucleotide (PCR-RSSO) and National Marrow Donor Program (NMDP) were used to analyze the polymorphism of HLA-A,B and DR alleles of 113 DMD patients and 406 normal controls in Han nationality of South China. RESULTS: The frequencies of HLA-A24, A30 alleles in DMD group were 11.25% and 5.46% respectively, indicating notable difference (P=0.001, < 0.01) from 22.16% and 0.87% of control group; the frequencies of HLA-B13, B15, B61 and B62 alleles in DMD group were 12.26%, 16.92%, 0.44% and 0.44%, indicating a notable difference (P=0.016, < 0.01, 0.001) from 6.76%, 1.49%, 4.79% and 5.05% of control group; the frequencies of HLA-DR04, DR07, DR12 alleles in DMD group were 17.45%, 6.40% and 19.62%, indicating a notable difference (P=0.018, < 0.01, 0.012) from 10.67%, 2.24% and 11.92% of control group. CONCLUSION: There are significant differences in the HLA gene frequencies between DMD patients and normal controls. These results suggest that HLA genotype relates to the muscular necrosis and the pathogenesis of DMD. PMID- 17922430 TI - [Cytogenetic and clinical study of myeloid leukemia]. AB - OBJECTIVE: To explore the clinical cytogenetic features and prognosis of myeloid leukemia patients. METHODS: Bone marrow direct method and/or 24h culture without phytohaemagglutimin(PHA) were used to prepare the chromosomes and karyotype analysis was performed with R-banding and G-banding techniques. RESULTS: Among 420 patients with acute myeloid leukemia (AML), 223 cases were found to exhibit clonal chromosome abnormalities, accounted for 53.1%. t(8; 21), t(15; 17), inv(16)and del(11) were specifically associated with M2b, M3, M4Eo and M5 respectively. Out of 158 patients with chronic myeloid leukemia (CML), 96.8% (153/158) were found to exhibit clonal chromosome abnormalities. T(9;22) was specifically associated with CML and some cases of M0, M1 and M2. In these myeloid leukemia cases, there were 18 cases (AML 13 cases, CML 15 cases) without clonal chromosome abnormalities, accounted for 3.1% (18/578) and this phenomenon agreed with the diagnose of clinical signs, marrow morphology and immunology incompletely. CONCLUSION: Karyotype analysis was not only helpful to the diagnose and differential diagnose of myeloid leukemia, but also an important standard of the remission, relapse and therapeutic effect of myeloid leukemia. Chromosome analysis can be made exactly with the probe and FISH technique on the basic of chromosome karyotype analysis. PMID- 17922432 TI - [Association of APOA5 gene polymorphism with levels of lipids and atherosclerotic cerebral infarction in Chinese]. AB - OBJECTIVE: To investigate the relationship between the polymorphism of apolipoprotein A5 gene (APOA5) -12238 T>C and atherosclerotic cerebral infarction (ACI). METHODS: Three hundred and forty-one subjects (170 ACI patients and 171 healthy controls) were collected to determine the genotypes by using polymerase chain reaction-restriction fragment length polymorphisms. RESULTS: APOA5 allele frequencies of T/C were 0.588/0.412 and 0.424/0.576 in ACI group and control group respectively. There was significant difference in allele and genotype frequencies between ACI group and control group (P < 0.05). The levels of plasma triglyceride in ACI patients with TT genotype were higher than those in patients with CC genotypes (P < 0.05). CONCLUSION: The relationship is found between the site of APOA5 gene -12238 T>C and ACI. There is a significant correlation between TT genotype of APOA5 and the levels of plasma triglyceride in patients with ACI. PMID- 17922431 TI - [The heat shock protein 70-2 gene +1267(A/G) polymorphism and susceptibility to ankylosing spondylitis in Chinese Han population]. AB - OBJECTIVE: To investigate the association between the heat shock protein 70-2 gene polymorphism and ankylosing spondylitis (AS). METHODS: The polymorphisms of HSP70-2 gene Pst I 1267 site were analysed in 176 Chinese Han AS patients and 127 healthy controls by PCR and restriction fragment length polymorphisms(RFLP) methods. RESULTS: In AS patients HSP70-2 genotypes AA, AG and GG were 46.6%, 46.0% and 7.4% respectively, frequencies of A and G were 69.6%(A) and 30.4%(G). In healthy controls HSP70-2 genotypes AA, AG and GG were 44.1%, 48.8% and 6.9% respectively, frequencies of A and G were 68.5%(A)and 31.5%(G). No significant differences were found in the distribution of HSP70-2 genotypes and allele frequencies between AS patients and controls. CONCLUSION: Our results indicate that the there may be no association of the HSP70-2 gene polymorphism with AS in Chinese Han population. PMID- 17922433 TI - [Microsatellite DNA polymorphism of adrenomedullin gene and atherosclerotic cerebral infarction]. AB - OBJECTIVE: To detect the correlation between the microsatellite DNA polymorphism of adrenomedullin(ADM) gene (repeated sequences of CA) and the atherosclerotic cerebral infarction (ACI). METHODS: With PCR, ADM genotype was monitored from 189 normotensive subjects and 283 cerebral infarction patients. By using radioimmunoassay, their plasma ADM concentration was measured, so as the biochemical index. RESULTS: The genotype distribution of ADM between the health control and ACI groups was significantly different, chi square was 28.732, P < 0.05. As one of the four alleles, including 11, 13, 14 and 19 alleles, the frequency of 19 allele in ACI groups was much higher than that in the health control group, chi square was 26.929, P < 0.05. However, there was no significant difference in plasma ADM concentration among the different genotypes of the ACI patients. CONCLUSION: Microsatellite DNA polymorphism of ADM gene may be associated with the genetic predisposition to ACI. PMID- 17922435 TI - [Polymorphism of the human platelet alloantigens HPA-3 and HPA-9w in the Chinese Han population]. AB - OBJECTIVE: To study the polymorphism of human platelet alloantigens HPA-3 and HPA 9w in the Chinese Han population. METHODS: A total of 1000 unrelated Chinese Han blood donors from different provinces of China were genotyped for HPA-3 and HPA 9w using PCR-sequence specific primer assay. RESULTS: Gene frequencies of 1000 Chinese Hans for HPA-3a and HPA-3b were 0.5935 and 0.4065 respectively, and all of them were HPA-9a positive. The distributions of HPA-3, HPA-9w of Chinese Hans which detected by chi-square criterion fit Hardy-Weinberg equilibrium. There were significant differences of the HPA-3 alleles gene frequency between Guangdong province and other five investigated provinces which included Shanxi, Heilongjiang, Zhejiang, Yunnan and Jiangsu. In comparison to other ethnic groups, no significant differences were observed in the distributions of HPA-3 except the Vietnamese and Australian. CONCLUSION: The results show that the chance of HPA-3 incompatibility were 0.3661 in random transfusion, and also provide a basis for researching on alloimmune thrombocytopenia and HPA-matched transfusion. PMID- 17922434 TI - [Association of -63A/C polymorphism of glutathione S-transferase M3 gene with essential hypertension in Chinese population]. AB - OBJECTIVE: To investigate the distribution of -63A/C polymorphism of human glutathione S-transferase M3(GSTM3) gene in Chinese Han population and the association of -63A/C polymorphism with essential hypertension (EH). METHODS: 63A/C polymorphism of GSTM3 gene was examined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) in 234 patients with EH and 328 healthy controls. The genotypes were confirmed partially by DNA sequencing. RESULTS: The distribution of GSTM3 -63A/C was in Hardy-Weinberg equilibrium. The CC genotype frequency in EH group (6%) was significantly higher than that in control group(1.8%) (P < 0.05). After adjustment for age, body mass index, and other risk factors, the CC genotype was independently associated with hypertension (OR=3.447, 95%CI: 1.19-7.63; P=0.04). CONCLUSION: The GSTM3 -63A/C polymorphism was associated with EH in Chinese Han population, the C allele might be a risk factor for EH in Chinese Han nationality. PMID- 17922436 TI - [Frequencies of GSTM1, GSTT1, and GSTP1 polymorphisms in ethnic minority from Guizhou]. AB - OBJECTIVE: To investigate the frequencies of GSTM1, GSTT1 and GSTP1 polymorphisms in Dong, Yi and Yao ethnic groups from Guizhou. METHODS: In 321 volunteers who were population-based, GSTM1 and GSTT1 polymorphisms were analyzed by a multiplex PCR procedure, whereas GSTP1 polymorphism was analyzed by PCR-RFLP method. RESULTS: Null genotype for GSTM1 and GSTT1 was 59.6%-71.2% and 39.4%-72.5%, respectively. The genotypic distribution of GSTP1 was 63.3%-75% for AA, 23.2% 35.8% for AG, 0-1.9% for GG, whereas the allelic frequencies were 81.2%-86.6% for the A allele, and 13.4%-18.8% for the G allele. CONCLUSION: There is a significant relationship between GSTT1 frequencies and ethnic populations. PMID- 17922437 TI - [Genetic polymorphisms of short tandem repeat loci D4S2368, D6S1043, D9S925 from Korean ethnic group of JiLin]. AB - OBJECTIVE: To understand the allele structure and genetic polymorphism at D4S2368, D6S1043, D9S925 short tandem repeat (STR) loci in Korean ethnic group of Jilin, and to construct a preliminary database. METHODS: The allele frequencies of the three STRs loci in 310 unrelated individuals from Korean ethnic individuals were analyzed by polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE). RESULTS: Seven, thirteen, and nine alleles were observed at D4S2368, D6S1043, and D9S925 loci, respectively, and all loci met Hardy-Weinberg equilibrium (except D6S1043). The statistical analysis of 3 STR loci showed the heterozygosities were more than 0.717, the polymorphic information contents (PIC) were more than 0.670; the combined power of discrimination (PD) and the power of exclusion (PE) were more than 0.9995 and 0.952 respectively. CONCLUSION: The three loci in this study are found to have high heterozygosity and polymorphic information content, so they can provide useful markers for genetic purposes. These results could serve as valuable data to enrich the Korean ethnic group genetic database and play an important role in Chinese population genetic application. PMID- 17922438 TI - [Genetic polymorphism of twelve Y chromosomal short tandem repeat loci in Chinese Hui ethnic group]. AB - OBJECTIVE: To obtain the genetic polymorphism of Y chromosomal short tandem repeat (Y-STR) loci in Ningxia Hui population. METHODS: Blood samples were collected from 150 unrelated healthy male individuals of Ningxia Hui ethnic group. Twelve Y-STR loci were amplified in one tube by using the PowerPlex System STR Amplification Kit, and the genotypes were determined using Genescan and Genotype software of ABI377 DNA sequencer and the frequency of alleles and haplotypes of Ningxia Hui ethnic was obtained. RESULTS: Seventy-five alleles were observed at 12 Y-STR loci. The frequency ranged from 0.0067-0.7067 and the gene diversity ranged from 0.4446-0.8877. Totally 148 different haplotypes were found, which were unique in 150 males. Two haplotypes were shared by 2 males respectively. The haplotype diversity was 0.9864. CONCLUSION: The 12 Y-STR loci are highly polymorphic in Ningxia Hui population and are suitable for genetics and forensic research. PMID- 17922439 TI - [Relationship between genetic polymorphism of MCP-1 and acute pancreatitis in Han population of Suzhou in China]. AB - OBJECTIVE: To study the relationship between monocyte chemoattractant protein-1 gene (MCP-1) -2518A/G polymorphism and acute pancreatitis (AP) in the Han population of Suzhou, China. METHODS: The polymorphisms were detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The genotypes and allele frequencies of MCP-1 -2518A/G were calculated and analyzed in 101 AP patients including 78 mild AP (MAP) patients and 23 severe AP (SAP) patients, and 120 healthy individuals as control group. RESULTS: The frequency of MCP-1 -2518 AA genotype in control group was significantly higher than that in SAP and MAP groups (P < 0.01). People with AG and GG genotypes had 5.896 times risk of developing MAP (P < 0.01, OR=5.896) compared with people with AA genotype. Subjects carrying G allele were at a 7-fold elevated risk for SAP (P < 0.05, OR=7.011) contrasted with subjects carrying AA genotype. However, no difference in AA genotypic distribution was noted between MAP and SAP groups (chi square=0.006, P=0.997). The frequency of G allele in healthy controls was obviously lower than that in MAP (P < 0.01, OR=0.318) and SAP groups (P < 0.01, OR=0.309). No difference of G allele frequency was found between SAP group and MAP group (P=0.623, OR=1.211). CONCLUSION: The MCP-1 -2518 AA genotype of the population in Suzhou may be a protective genotype of AP. People with higher frequency of G allele is more likely to suffer from AP. Nonetheless, the genotype of AA and the frequency of G allele couldn't predict the risk of SAP. PMID- 17922440 TI - [The chronic renal failure epidemic: an underestimated public health problem]. AB - Renal failure is considered a rare disease. However, recent epidemiological surveys like the NHANES III survey in the USA have shown that mild and moderate renal insufficiency is much more common: 31% and 4% of Americans, respectively, are affected by a mild or moderate degree of renal insufficiency. Such an epidemic is of particular concern because of the high cardiovascular risk brought about by kidney failure. Renal insufficiency is now considered a public health priority. Together with diabetes and smoking, the metabolic syndrome is the principal factor responsible for this epidemic. The prevalence of chronic renal insufficiency is in fact strictly proportional to the number of components of the metabolic syndrome being present in individual patients. As renal function deteriorates, other risk factors come into play like those peculiar to renal insufficiency (anemia, hyperparathyroidism) and some so-called emerging risk factors (inflammation, hyperhomocysteinemia and high plasma levels of endogenous inhibitors of NO synthase such as asymmetric dimethylarginine). PMID- 17922441 TI - [Dyslipidemia and the risk of kidney disease]. AB - An abnormal lipid profile is very frequent in patients with kidney disease due to the well-known nephrotoxicity of lipids. During progression of chronic kidney disease, the excretion of triglycerides, LDL and proteins increases while the glomerular filtration rate declines. Blood lipoproteins and lipids are modulated depending on the type of treatment: hemodialysis, CAPD or renal transplant. We analyzed many studies on dyslipidemia in patients with kidney disease by comparing different therapies. The use of statins reduces protein excretion and hyperlipidemia as well as progression of chronic renal failure with a direct effect on mesangial cell proliferation. A decrease in total cholesterol and LDL occurs in hemodialysis patients, a decrease in LDL and an increase in HDL occur in CAPD patients, and a decrease in LDL and triglycerides is observed in renal transplant recipients; in the latter, graft survival increases without there being any relevant correlation with immunosuppressive treatment. In conclusion, we found that statins are useful to contrast the progression of chronic kidney disease and atherosclerosis in hemodialysis and CAPD patients and to reduce chronic allograft nephropathy in renal transplant recipients. PMID- 17922443 TI - [Pathophysiology and prevention of contrast-induced acute renal failure]. AB - Although the infusion of iodinated contrast media in diagnostic and interventional procedures may cause acute renal failure (ARF) especially in older or diabetic patients with preexisting nephropathy, these procedures are often unavoidable. Contrast medium-induced ARF is defined as an increase in serum creatinine of 0.5 mg/dL or a 25% or greater relative increase from baseline within 72 hours of iodinated contrast medium infusion. Because it is often very difficult to employ alternative diagnostic procedures, it is mandatory to adopt prophylactic protocols to prevent radiocontrast nephropathy. Renal hemodynamic lesions leading to medullary hypoxia, oxygen free radicals inducing tubular cell alterations, and parenchymal vasoconstriction are the main factors in the pathogenesis of contrast-induced ARF. Among the many proposed protocols to prevent contrast-induced renal toxicity, the most effective procedure is hydration with 1 mL/kg/h of isotonic saline solution in the 12 hours before and after contrast medium infusion. Promising results in terms of cardiac and renal protection have been reported in a recent trial with the use of high-dose N acetylcysteine acting as an oxygen free radical scavenger: an intravenous bolus of 1200 mg N-acetylcysteine was given before coronary angiography followed by 1200 mg orally twice a day for 48 hours after the procedure. The protective effect seemed to involve not only the kidney: the drug was found to induce a significant reduction of the necrotic area in myocardial infarction. PMID- 17922442 TI - [Risk and prevention of diabetic nephropathy]. AB - Diabetic nephropathy is one of the most frequent causes of end-stage renal disease (ESRD), and there has been a dramatic increase in the number of patients entering renal replacement therapy in the last few years. Moreover, diabetic nephropathy is associated with elevated cardiovascular morbidity and mortality. Prevention and treatment of diabetic nephropathy is based on optimal metabolic and blood pressure control, proteinuria reduction, and renin-angiotensin aldosterone system (RAAS) inhibition. In the normoalbuminuric patient, optimal glycemic control (HbA1c below 7.0%) plays a fundamental role in the primary prevention of ESRD. Furthermore, blood pressure levels below 130/80 mmHg are strongly recommended. In the microalbuminuric stage, strict glycemic control (HbA1c below 7.0%) likely reduces the incidence of overt nephropathy, while blood pressure values less than 130/80 mmHg are recommended. Moreover, there is evidence that inhibition of RAAS, either by angiotensin-converting-enzyme inhibitors (ACE-I) or angiotensin-receptor blockers (ARB), reduces the development of overt nephropathy, regardless of the blood pressure levels. ACE-I are recommended as the drugs of choice in type 1 diabetes, while both ACE-I and ARB are considered first-choice drugs in type 2 diabetes. Once overt proteinuria has developed, it is uncertain whether glycemic control affects the progression of nephropathy, which is strongly influenced by blood pressure and proteinuria. Optimal blood pressure levels are < 130/80 mmHg in patients with proteinuria < 1 g/day and < 120/75 mmHg in patients with proteinuria > or =1 g/day. In type 1 diabetes there is consensus on the renoprotective role of ACE-I, while in type 2 diabetes, ARB have been shown to be more effective than conventional therapy or calcium-channel blockers in slowing the progression of nephropathy. Lastly, a multifactorial therapeutic approach based on optimal glycemic control, intensive antihypertensive therapy, inhibition of RAAS, statins and aspirin is pivotal in the prevention and treatment of diabetic nephropathy. PMID- 17922444 TI - [Recent advances in the prevention of cardiovascular morbidity and mortality in end-stage renal disease: role of anemia, hyperparathyroidism and calcifications]. AB - The mortality rate in patients with end-stage renal disease (ESRD) is extremely high, mainly because of the high prevalence of cardiovascular disease. In addition to traditional cardiovascular risk factors, other factors peculiar to chronic kidney disease play a role. Anemia and calcium-phosphate disorders are of particular interest, not only because they have been related to an increased risk of death but, more importantly, because they can be reversed by treatment, thereby providing the opportunity to prevent or delay the onset of cardiovascular disease. Despite a clear association between higher hemoglobin levels and better survival, data from interventional trials do not seem to show a significant positive effect of hemoglobin normalization with erythropoiesis-stimulating agents on survival and left ventricular mass in ESRD patients. Nevertheless, partial correction of anemia is still an important goal to be reached, as is also suggested by international guidelines. Disorders of calcium-phosphate metabolism have also been clearly related to increased mortality. Unlike anemia, which can be easily corrected by treatment in most cases, mineral metabolism is much less effectively treated. New agents, such as phosphate binders not containing calcium and aluminum, vitamin D analogs with lower calcemic activity, and calcimimetics, are becoming increasingly available in everyday clinical practice and are likely to allow a higher percentage of patients to achieve the recommended targets for calcium-phosphate and parathyroid hormone. Given that these molecules have only been introduced recently, clear data from interventional studies showing improved survival after adequate correction of mineral metabolism parameters are still lacking. PMID- 17922445 TI - [Prevention of nosocomial infections: surveillance based on microbiological data]. AB - BACKGROUND: In the C. Poma Hospital of Mantua we have been using a system of continuous surveillance of nosocomial infections based on microbiological data for the past 4 years. This monitoring estimates the incidence of the microorganisms found in cultures, especially those at risk of causing nosocomial infections. MATERIALS AND METHODS: Since June 2001 microbiological data have been registered using the Mercurio-Dianoema software and elaborated by means of Microsoft Excel in order to obtain information about isolated bacteria, especially those resistant to antibiotics. RESULTS: Surveillance in "critical" wards revealed the presence of Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans in the intensive care unit in the period 2003-2005. The most frequent bacteria in hemodialysis have been coagulase-negative Staphylococci and Staphylococcus aureus, with variable methicillin resistance. CONCLUSION: The analysis of microbiological data has promoted effective measures to reduce the incidence of these bacteria (increased rules of good practice, hand washing, etc.). If nosocomial infections or high-risk microorganisms occur, assessments are carried out; monitoring of the antibiotic resistance of the bacteria is very important. PMID- 17922446 TI - [Outpatient monitoring after renal transplantation: protocols shared between transplant center and local nephrology units]. AB - Outpatient monitoring plays a key role in the long-term success of kidney transplantation. Shared management of transplanted patients between transplant centers and local nephrology units is becoming common practice and is a benefit both for the patients, who can be followed in an outpatient office closer to their homes, and for the transplant centers, which are overwhelmed by an increasing number of follow-up patients. The program is also well accepted by the referring nephrology units, which are interested in improving their skills. In this article a model of clinical collaboration is discussed, although it is well known that it is impossible to apply the same rules to all centers. However, to make any collaborative program feasible, two main requirements must be met. First, every local unit should have a referent nephrologist responsible for the clinical follow-up of transplant recipients and for the waiting list; second, every transplant center should organize transplant refresher courses for their referring nephrologists. PMID- 17922447 TI - [Low-toxicity immunosuppressive therapy in renal transplant]. AB - INTRODUCTION: Renal allograft loss in the long term may be due to the death of a patient with a functioning graft or to chronic allograft nephropathy. One of the most important factors in the development of chronic allograft nephropathy is drug nephrotoxicity. The term nephrotoxicity comprises two distinct forms of renal injury: acute and chronic. Immunosuppressive drugs, and in particular calcineurin inhibitors, have a variety of side effects including nephrotoxicity. The nephrotoxicity associated with calcineurin inhibitors is well known; this association has also been described for the newer agents. METHODS: We reviewed a large number of recent studies that attempted to reduce the toxicity of immunosuppressive regimens. RESULTS: A number of low-toxicity protocols have been developed. Encouraging results have been obtained with regimens that reduce or eliminate nephrotoxicity-inducing calcineurin inhibitors and with regimens that reduce or eliminate steroids, which are responsible for many diseases that may lead to the death of the patient, even with a functioning graft. CONCLUSION: All immunosuppressive drugs may be nephrotoxic, even if they act through different mechanisms. Combining different drugs at low dosage would therefore seem the best solution. It is not yet clear which regimens will be the most effective from the point of view of maximizing patient and graft survival, minimizing rejection, and minimizing adverse events. PMID- 17922448 TI - [From evidence to practice: a difficult but not impossible itinerary]. AB - In recent years, evidence-based medicine (EBM) has acquired its own dignity and cultural identity, and increasing importance. EBM helps doctors understand that the uncertainties of medicine must be analyzed quantitatively in order to rationalize and systematize the information gathered from clinical observation; on the other hand, it provides them with suitable instruments to estimate the performance of diagnostic tests and the efficacy of drugs. One of the main merits of EBM has been the progressive spread of randomized controlled trials as the gold standard for evaluating the effectiveness of treatments. EBM's primary objectives can be summarized by the following points: 1) To transform the physicians' need for information into questions that may be answered (ie, formulate the questions); 2) To find in an as efficient way as possible the best evidence to answer these questions; 3) To critically evaluate the evidence obtained (ie, assign a weight to it) in order to determine its validity (ie, its approximation to the truth) and its usefulness (ie, its concrete clinical applicability); 4) To introduce into clinical practice the conclusions drawn from the results; 5) To estimate individual physicians' performance (ie, one's own conduct and efficiency). EBM's advantages are not only that physicians have acquired a method to search for the right evidence and to apply diagnostic and therapeutic procedures, but, more importantly, that it has provided them with the only tool for true quality improvement, namely the critical appraisal of their own work. Unfortunately, the latter is too often based on a different type of EBM: Evidence-Based Medicine. PMID- 17922449 TI - [Continuing education aimed at occupational risk prevention]. AB - The implementation of European Commission directives has radically modified occupational risk management. In fact, widespread and continuous education addressed to exposed workers is seen by the legislator as one of the most effective means to avoid occupational diseases. The recent establishment of Continuing Medical Education (CME) has helped the system grant CME credits to participants in occupational courses organized by health-care providers. Moreover, the constant development of new health technologies coupled with the high productivity of the legislator confer a short lifetime on the acquired knowledge. Biological risks, biomechanical overload of the lumbar spine, and stress are among the classical risks requiring attention, discussion, and regular updating. Not only health-care workers but also safety technicians, occupational physicians, managers and union representatives should receive scheduled refresher training. Modern technologies such as distance learning, interactive simulation software, and online training tools are often the best teaching solutions. Occupational disease prevention should no longer be seen as a cost but as a cost savings indicator. Since it is closely related to quality promotion, it should play a primary role in the core business of health-care providers. PMID- 17922450 TI - [Professional and legal responsibilities of nurses]. AB - The responsibility of nursing personnel is characterized by civil, criminal law, and disciplinary aspects. The causal relationship between a certain behavior and the resulting harmful event and the methodology of assessment of events as rationally foreseeable -- case by case and not only in statistical terms -- are elements to be taken into account. The question of nursing responsibility must be placed in the context of the competence and autonomy of the nurse in the nurse patient alliance and in the context of nurses' professional and educational profiles. The relationship between doctor and nurse and nurse and medical social worker in the diagnostic and curative course are elements for reflection. PMID- 17922451 TI - [Nurse's coworking to electronic medical record]. AB - Nephrologists need to register and look at a great number of clinical data. The use of electronic medical records may improve efficiency and reduce errors. Aim of our work is to report the experience of Villa Scassi Hospital in Genoa, where a "patient file" has been performed to improve nephrology practice management. The file contains all clinical records, laboratory and radiology data, therapy, dialysis clinics, in addition to reports of out-patients department. This system allowed a better efficiency in diagnosis and treatment of the patient. Moreover experience of nurses in employing electronic medical records is reported. A reduced number of errors was found in therapy administering, because of a only one data source for physicians and nurses. PMID- 17922452 TI - [Acute renal failure after videolaparoscopic surgery: an avoidable complication?]. AB - Videolaparoscopic surgery exposes the abdominal organs to the mechanical effect of pneumoperitoneum at pressure values between 12 and 15 mm Hg, which are considered safe. Nevertheless, experimental data have shown that this pressure range can represent a hemodynamic risk factor as it may induce a decrease in the venous return to the right ventricle, a decrease in cardiac output, and activation of the sympathetic nervous system and renin angiotensin system. We report two cases of acute renal failure that occurred soon after videolaparoscopy in young female patients without any evidence of ongoing renal disease. Patient A was 29 years old and was submitted to videolaparoscopic surgery in a follow-up program after surgical treatment of ovarian cancer; patient B was 15 years old and was submitted to the surgical removal of a monolateral ovarian cyst. In neither of the cases was it necessary to perform hemodialysis. Patient A underwent a renal biopsy under ultrasound guidance; optic microscopy showed only in ra- and extraglomerular capillary congestion. In both cases the acute renal failure resolved completely and the patients where discharged with normal renal function. Taking in to account that normal renal venous pressure levels are around 4 mmHg we think that a) a 15 mmHg pneumoperitoneum may represent a risk factor during videolaparoscopic surgery mainly if the patient's extracellular volume is not properly expanded; b) administration of nonsteroidal anti inflammatory drugs in order to prevent surgical pain may inhibit vasodilatory prostaglandin availability; c) onset of oliguria during the surgical procedure suggests that extracellular volume expansion is required. PMID- 17922453 TI - ["Terzo fuoco", lead poisoning and chronic renal failure]. AB - BACKGROUND: In ceramics, "Terzo fuoco" (Third fire) means a third firing of clay to fix colors on tiles or pottery. The low firing temperatures (800-900 degrees C) and the use of a spray gun are risk factors for lead poisoning. Because of their small size, handicraft companies often fail to implement the preventive measures that are utilized efficiently in large tile factories. We report a case of chronic lead poisoning with special emphasis on diagnostic tools and treatment goals. CASE REPORT: A 38-year-old woman was hospitalized in 2005 because of grade 3 chronic renal failure (serum creatinine 1.69 mg%, Cockroft-Gault glomerular filtration rate [GFR] 45 mL/min), hypertension and elevated serum uric acid (13.4 mg%) without gout. She had been previously hospitalized elsewhere and diagnosed as suffering from hypertensive nephropathy. Her occupational history included acute lead poisoning 12 years before, which was treated with a short leave from work. She subsequently continued her job, using a spray gun for decorative drawing in a small tile company until 2004. Because of a low GFR she underwent a 3-day chelation test with 750 mg CaNaEDTA i.v., and excreted 1056 microg Pb (n.v < 600 microg) -- (PbU/EDTA ratio 1.41; n.v < 0.6). A renal biopsy showed chronic interstitial nephropathy with severe arteriolosclerosis. The patient was treated with 5 courses of EDTA, resulting in a final Pb excretion of 517 microg/72 h (PbU/EDTA 0.69). Her serum creatinine fell to 1.32 mg% (CFR 58 mL/min). A further course of chelation therapy is planned. DISCUSSION AND CONCLUSIONS: The EDTA chelation test allows to determine the lead body burden (PbBB) and to titrate subsequent chelation therapy. Recent papers have shown that PbBB is a major factor in the progression of chronic renal failure besides pressure control, and have indicated a PbBB safety level of less than 100 microg/test (PbU/EDTA < 0.1). In order to prevent the development of chronic renal failure, it is important that not only occupational but also environmental lead exposure be identified and adequately treated. PMID- 17922454 TI - [Early diagnosis and prevention of renal disease: collaboration between nephrologists and general practitioners]. AB - Our outpatient clinic activity has taught us that a working relationship between general practitioners (GPs) and nephrologists may improve the definition of the diagnostic-therapeutic course for the benefit of the patient. We have therefore contacted the 7 teams comprising 104 GPs and pediatricians working in the area of the Agnelli Hospital in Pinerolo (132,000 inhabitants in 1,404 square kilometers) to assess the possibility of improving and strengthening the collaboration between GPs and nephrologists. The starting point was a direct telephone link aimed at dealing with patients' kidney problems in real time, evaluating history and clinical data, establishing the best timing of treatment, and defining the diagnostic and therapeutic options. The initiative was welcomed at all team meetings and it stimulated further requests for collaboration. One of the main requests was for simple clinical guidelines to deal with the most frequent clinical nephrological issues. This is the program we are carrying out: 1) We have established consulting hours during which GPs can call nephrologists at the hospital to discuss the best diagnostic-therapeutic approach for individual kidney patients. 2) We have identified diseases of common interest (isolated urinary abnormalities; hypertension; nephrotoxicity; abnormal renal function; chronic renal failure; urinary infections; kidney stones). 3) We have planned to draw up clinical guidelines. 4) We will discuss each draft with the team of GPs. On the basis of the gathered suggestions, we will prepare a final version of the guidelines to be sent to the GPs and pediatricians of our area. PMID- 17922455 TI - [Hospital-primary care teamwork in nephrology: 'preventive' epidemiology of diabetic nephropathy]. AB - BACKGROUND: The onset of clinical proteinuria in patients with diabetic nephropathy usually marks the existence of irreversible and progressive kidney damage. Prevention of chronic renal failure ought to take place in earlier stages, mostly in the outpatient setting and in close collaboration with general practitioners (GPs). This study aims to evaluate the prevalence and the clinical stage of diabetic nephropathy and to screen for progression factors in the regional community referred to the outpatient department of the nephrology unit of the Mons. DiMiccoli Hospital in Barletta; it is intended as the first phase of a collaborative project for the prevention of diabetic nephropathy. METHODS: Our team asked for the collaboration of GPs within the geographic area referring to our outpatient department. This led to the institution of a working group aimed at interrelating primary with hospital care in the setting of nephrology. The clinical data of diabetic patients as well as their codified personal data were classified by GPs and subsequently processed by the nephrology team. The glomerular filtration rate was estimated by means of the abbreviated MDRD study equation. Finally, the cumulative prevalence of known risk factors for diabetic nephropathy progression was assessed in a subgroup of 201 diabetic patients. RESULTS: Within the whole group of screened subjects from the regional community (21,314), the prevalence of diabetes was 3.54%, and the rates of personal and clinical features did not show any substantial differences from the ones recorded in Italy as a whole. In the subgroup of 201 diabetic patients, the cumulative frequency of progression factors for diabetic nephropathy showed a relevant percentage of subjects with clinical and laboratory features deviating from the targets proposed by current guidelines, thus justifying the adoption of specific prevention plans. CONCLUSIONS: This study shows the feasibility and the advantages of setting up a stable collaboration between hospital and primary care in order to improve the diagnosis and prevention of kidney disease. Identification of the most frequently deviant progression factors provides the primary target for the prevention of kidney damage in diabetic patients. PMID- 17922456 TI - [Evaluation of outpatient referral to the nephrologist]. AB - BACKGROUND: Late referral to the nephrologist has been mainly examined to assess its causes and effects in patients with chronic renal failure. It has not been analyzed as a phenomenon in itself, to identify its features and allow more effective prevention plans. This study proposes a methodological approach to the study of outpatient referral to the nephrologist (ORN) by defining a number of evaluation tools. METHODS: Patients referred to an outpatient nephrology department represent a cohort of selected subjects on the basis of patient related factors (clinical, social and cultural) and care-related factors. The incidence and prevalence of kidney disease in this population cannot be considered as a measure of the frequency in the general population. They may represent a first-level approach to the study of ORN by providing an indication of its pattern but not of its actual size in the catchment area. For this purpose we implemented a "referral index", calculated as the percent ratio between the number of affected subjects attending our outpatient department and the number of affected subjects within the whole catchment area. RESULTS: The number of subjects attending our outpatient department showed a progressive increase, especially among diabetics, in the last 8 years, in concomitance with a collaborative prevention project involving general practitioners. The referral index of the most frequent chronic kidney diseases in 2005 never exceeded 4.9%, accounting for the steady increase in outpatients following our ORN promotion policy. Prevalence on the one hand and referral index on the other provide different types of information about the same subgroup of outpatients: among patients with chronic kidney diseases, prevalence shows a different graduation of ORN than the referral index. Finally, the frequency distribution of chronic kidney disease stages calculated with the referral index moved toward the fourth and fifth stages rather than the second and third stages, as shown by the prevalence. CONCLUSIONS: The incidence and prevalence of renal diseases among outpatients are measures of ORN and they are useful for a qualitative analysis, such as to assess its pattern. The referral index provides a quantitative evaluation of ORN with respect to the general population of the catchment area. PMID- 17922457 TI - [Development of extracorporeal blood purification methods: Molecular Adsorbent Recirculating System (MARS) for hepatic and renal function replacement]. AB - The Molecular Adsorbent Recirculating System (MARS) clears the blood from catabolites that either occur free in the plasma water (through dialysis), such as uremic toxins and ammonia, or are bound by albumin, such as hepatic toxins. The latter are transferred from the albumin in the blood to the albumin circulating in a closed loop where toxins are removed by adsorption on resins (charcoal and ion-exchange resin). The efficacy of this extracorporeal blood purification method in the treatment of acute or acute-on-chronic liver failure (also associated with renal failure) has been demonstrated in numerous studies. Fifty-one patients, 5 affected by acute liver failure and 46 by acute-onchronic liver failure (8 of them with additional renal failure) were treated with MARS. The results demonstrated that the method, which effectively removes ammonia, bilirubin, bile acids and uremic toxins, reduces the blood concentration of these molecules. It thereby improves the patient's clinical condition and biochemical parameters including cholinesterase, alkaline phosphatase and prothrombin activity, eliminating, in addition, the drug-refractory pruritus that is a very frequent symptom in cholestatic liver disease. These results agree with those reported in the literature concerning the efficacy of MARS in the replacement of the detoxifying function of kidneys and liver. PMID- 17922458 TI - Development of ELISAs for quantification of HMFG1-specific human anti-mouse IgG and IgM antibodies. AB - The aim of this study was to develop and validate ELISAs for quantification of HAMA-IgM and HAMA-IgG in serum of patients with ovarian cancer who enrolled in a large international randomized phase III trial of intraperitoneal Yttrium-90 labeled HMFG1 murine monoclonal antibody therapy. The capture antibody of these 2 assays was the murine antibody HMFG1, while mouse anti-human IgM-HRP or mouse anti-human IgG(Fc)-HRP served as tracer antibodies. A pool of HAMA-positive serum samples was used to prepare a series of assay standards and another pool served as reference preparation. The analytical sensitivity of the HAMA-IgM assay was 2.5 arbitrary units per mL (AU/mL) and 4.7 AU/mL for the HAMA-IgG ELISA. Diluted serum samples showed good parallelism with the HAMA-IgM and HAMA-IgG standard dose-response curves. Within-assay coefficient of variation was 7.5% for HAMA-IgM and 6.5% for HAMA-IgG. Between-assay variation was 14.2% for HAMA-IgM and 15.3% for HAMA-IgG. The developed HAMA-IgM and HAMA-IgG ELISAs show satisfactory reliability criteria (sensitivity, parallelism and precision) and are suitable for monitoring of HAMA-IgM and HAMA-IgG responses in ovarian cancer patients. These ELISAs will be used to monitor the development of HAMAs in patients who received radioimmunotherapy with murine HMFG1. PMID- 17922459 TI - Peritoneal fluid cytokines and the differential diagnosis of benign and malignant ovarian tumors and residual/recurrent disease examination. AB - This study aimed to assess the potential value of peritoneal fluid cytokine examination for the differential diagnosis of ovarian tumors and for evaluating residual or recurrent disease after treatment. The cytokines that are commonly elevated in ovarian cancer, VEGF, IL-6, bFGF, IL-8 and M-CSF, and a reference ovarian tumor marker, CA 125, were measured in peritoneal fluids of 53 previously untreated patients with epithelial ovarian cancer, 18 ovarian cancer patients after surgical treatment and chemotherapy, and 17 patients with benign epithelial ovarian tumors. Non-parametric statistical analysis of data was performed. Ovarian cancer peritoneal fluids, as compared to peritoneal fluids of patients with benign ovarian tumors, contained significantly higher concentrations of IL 6, VEGF and CA 125, and significantly lower concentrations of bFGF and M-CSF, but only the levels of IL-6 and VEGF were significantly higher in peritoneal fluids of stage I and II ovarian cancer patients than of patients with benign ovarian conditions. IL-6 at the cutoff level of 400 pg/mL discriminated benign and malignant ovarian tumors with 92% sensitivity and 60% specificity, while VEGF at the cutoff of 400 pg/mL had 90% sensitivity and 80% specificity. At the cutoff level of 1200 pg/mL, IL-6 had 84% sensitivity and 87% specificity. A radical decrease in local cytokine and CA 125 levels in patients after treatment was independent of therapy outcome. IL-6 and VEGF measurements in peritoneal fluids might be useful for the differential diagnosis of malignant and benign ovarian conditions, but not for residual or recurrent disease examination. PMID- 17922461 TI - Significant changes in circulating plasma levels of IGF1 and IGFBP3 after conventional or dose-intensified adjuvant treatment of breast cancer patients with one to three positive lymph nodes. AB - The insulin-like growth factor 1 (IGF1) and its binding protein IGFBP3 (insulin like growth factor binding protein 3) play a pivotal role during the growth and development of tissues. The purpose of this study was to evaluate the influence of anthracycline- and taxane-containing adjuvant chemotherapy in breast cancer patients on the circulating plasma levels of IGF1 and its main binding protein, IGFBP3. This investigation was part of a prospective randomized phase III study in which breast cancer patients were treated with either conventional or dose intensified adjuvant chemotherapy. The factors were quantified in the plasma of 151 patients with a commercially available sandwich enzyme immunoassay. Before therapy, both parameters were within the normal range in most patients (n=145 and n=144). After therapy, both factors had increased significantly by 29% (IGF1) and 19% (IGFBP3), with the highest increase being observed in the dose-intensified group. Correlations with patient and tumor characteristics revealed a relatively higher increase in both parameters in premenopausal patients, patients with lower grade tumors, more positive lymph nodes, larger tumor volume, and positive hormone receptor status. No correlation was found with the HER2 expression of the tumors. PMID- 17922460 TI - Nuclear and cytoplasmic expression of ErbB-4 in prostate cancer. AB - PURPOSE: To evaluate cytoplasmic and nuclear ErbB-4 expression in prostate cancer specimens and its association with outcome. BASIC PROCEDURES: Specimens of 50 prostate cancer patients were investigated for ErbB-4 overexpression using Immunohistochemistry staining. Cytoplasmic and nuclear staining was graded as 0-3 according to its intensity. The prognostic parameters were tumor stage, PSA level, Gleason score, probability of positive lymph nodes (Partin's tables and Roach equation), and 5-year disease free survival (Kattan nomogram). MAIN FINDINGS: Overexpression of ErbB-4 (> or = 1) was detected in 30 (60%) patients and overexpression using cytoplasmic and nuclear staining was > or = 2 in 19 (38%) and 17 (34%) patients, respectively. In only one third of the specimens was there any similarity between the 2 types of staining. Advanced tumor stage, high pretreatment PSA levels and high Gleason scores were evenly distributed among the patients with low (< or = 1) and intermediate/high (> or = 2) ErbB-4 expression. The probability of lymph node involvement and 5-year disease free survival were similar in both types of staining. PRINCIPAL CONCLUSIONS: ErbB-4 was overexpressed (cytoplasmic and nuclear staining) in approximately one third of prostate cancer patients. The rate of similarity between the 2 staining types was only 33%: overexpression was evenly distributed among intermediate/high and low risk prostate cancer patients with both staining methods. PMID- 17922463 TI - Evaluation of serum CA 125 levels in patients with pelvic pain related to endometriosis. AB - The aim of the study was to investigate the clinical value of the serum CA 125 level for diagnosing and determining the severity of endometriosis and pelvic pain associated with endometriosis. Eighty-six women who underwent operative laparoscopy were enrolled. Sixty-nine women with endometriosis and 17 without endometriosis participated in this study. In all of the patients, endometriosis was diagnosed and classified into stages according to the Revised American Fertility Society (R-AFS) classification. The mean serum CA 125 levels were determined in each patient. We also investigated the relationship between serum CA 125 concentration and the intensity of dysmenorrhea and dyspareunia in the study group. The mean serum CA 125 levels of women with endometriosis were higher than those of the control group (p<0.050). However, the mean serum CA 125 levels were higher in stage IV than in other stages of endometriosis according to the R AFS classification. On the other hand, the percentage of patients with serum CA 125 levels >35 U/mL was elevated in the subgroups with severe dyspareunia and severe dysmenorrhea versus the asymptomatic subgroup but the differences had no statistical significance. In conclusion, CA 125 serum levels were related to endometriosis and R-AFS score in the evaluated patient series. No correlation was found between serum levels of CA 125 and pelvic pain in patients with endometriosis. PMID- 17922462 TI - Use of plasma Epstein-Barr virus DNA monitoring as a tumor marker in follow-up of patients with nasopharyngeal carcinoma: preliminary results and report of two cases. AB - Recent studies suggest that plasma Epstein-Barr virus (EBV) DNA may reflect tumor burden in patients with nasopharyngeal cancer. A prospective study was initiated to investigate this correlation in 125 patients (34 pretreatment [Group A], 78 in remission [Group B] and 13 relapsed [Group C]) and 19 healthy controls. In group A, EBV DNA was detected in plasma samples of 24 (70%) patients. In Group B, EBV DNA was detected in 7 patients (range 77-13,731 copies/mL) and further imaging in all but one of these patients revealed active disease confirmed by ultrasound guided fine-needle biopsy. There was only one false-positive case; this patient is currently under follow-up. Here we describe 2 of the 7 patients with detectable plasma EBV DNA in whom recurrence was documented by PET scan during follow-up. Our results showed that in group B the positive predictive value of quantitative analysis of plasma EBV DNA was 85%. Quantitative analysis of EBV DNA in plasma seems to become an integral part of screening, staging, monitoring, and prediction of relapse in patients with nasopharyngeal carcinoma. However, previous studies cannot be considered definitive and more reports on the use of this technique are urgently needed from both endemic and non-endemic regions. PMID- 17922465 TI - Five-year stability study of free and total prostate-specific antigen concentrations in serum specimens collected and stored at -70 degrees C or less. AB - The stability of total (t) and free (f) prostate-specific antigen (PSA) in male serum specimens stored at -70 degrees C or lower temperature for 4.7 to 4.9 years was studied. Until now, the stability of these analytes in serum has not been evaluated systematically beyond 2 years of storage at -70 degrees C. Aliquots of frozen serum were thawed in 2001 and 2006 and assayed for tPSA and fPSA using a Dade Behring Dimension(R) RxL analyzer and reagents. tPSA values ranged from 0.07 to 69.94 and 0.00 to 69.83 ng/mL in 2001 and 2006, respectively, whereas fPSA values for the tested specimens ranged from 0.02 to 5.72 and 0.00 to 5.92, respectively. Deming regression analyses showed agreement in assay values over time as tPSA values yielded a slope of 1.0112 and a y-intercept of 0.0195; fPSA values produced a slope 1.0538 and a y-intercept of -0.0442; f/tPSA values yielded a slope of 0.9631 and a y-intercept of 0.1195. A Bland-Altman analysis of the data demonstrated analyte and ratio stability over this time period. We conclude that serum, when collected properly and stored at -70 degrees C or lower temperature, may be used for tPSA and fPSA clinical studies for at least 5 years after collection. PMID- 17922464 TI - Elevation of serum CA 125 and D-dimer levels associated with rupture of ovarian endometrioma. AB - BACKGROUND: Patients with endometriosis rarely have a serum CA 125 concentration >100 IU/mL. A raised plasma level of D-dimer indicates active fibrinolysis, either secondary to clot formation or primarily activated. This condition is seldom diagnosed in patients with endometriosis. CASE REPORT: A 53-year-old woman was referred to our institution for acute abdominal pain. Laparoscopic surgery revealed a large ovarian cyst with rupture on the left side. Preoperative laboratory tests detected high serum CA 125 and D-dimer levels. Adnexectomy was performed, resulting in a sharp decrease in serum CA 125 and D-dimer concentration. We describe the clinical course of the patient. CONCLUSION: Rupture of a large ovarian endometrioma can lead to a high serum concentration of CA 125, a condition which, in addition to the detected pelvic mass, may mimic a malignant process. The increased D-dimer plasma level indicated that a ruptured endometriotic cyst can induce coagulation reactions. PMID- 17922466 TI - Beta-CTX and ICTP act as indicators of skeletal metastasis status in male patients with non-small cell lung cancer. AB - Bone metastasis is common in lung cancer patients and associated with reduced quality of life and reduced overall and median survival, so the early detection of bone metastasis and monitoring of its status is very important for clinicians. Serum bone-specific alkaline phosphatase (BAP), osteocalcin (OC), beta isomer of C-terminal telopeptide of type I collagen (beta-CTX) and cross-linked C-terminal telopeptide of type I collagen (ICTP) were compared with regard to their usefulness as indicators of bone metastasis in lung cancer. The serum concentrations of the 4 markers were measured by commercially available tests in 96 male patients with non-small cell lung cancer and 30 male patients with other pulmonary diseases. The levels of both a-CTX and ICTP were significantly higher in 61 lung cancer patients with bone metastases than in 35 lung cancer patients without bone metastases (both p<0.001), and significantly correlated with the extent of bone disease. Although ICTP had a better sensitivity and accuracy than beta-CTX (75.4% vs 65.6% and 72.9% vs 68.8%, respectively), they had a similar area under the receiver operating characteristic curve (0.85 vs 0.83). These results support the use of beta-CTX and ICTP as an adjunct tool for the diagnosis and screening of bone metastasis in lung cancer. PMID- 17922467 TI - High expression of nuclear factor of activated T cells in Chinese primary non small cell lung cancer tissues. AB - PURPOSE: Nuclear factor of activated T cells (NFAT) has been reported to be involved in the development of various types of cancer including adenocarcinoma of the breast. This research was the first to investigate NFAT protein expression in primary non-small cell lung cancer (NSCLC) tissues from Chinese patients. METHODS: NFAT protein expression was determined in 130 surgically resected primary NSCLC and matched normal tissues by immunohistochemical analysis. The association between NFAT expression and clinical categorical variables was further analyzed with the SPSS software. RESULTS: We found that NFAT expression was much higher in 85 tumor tissues (65.4%) and lower in 45 tumor tissues (34.6%) compared with the matched normal tissues. Further statistical analysis by the chi square test showed that high expression of NFAT proteins was significantly associated with tumor differentiation (p=0.045), invasion (p=0.031), histology (p<0.0001), tumor size (p=0.038) and cigarette smoking history (p=0.024). However, there was no correlation between the expression of NFAT proteins and pTNM classification, and no difference in 5-year survival rate between patients with high or low expression of NFAT proteins. Multivariate logistic regression analysis for the correlation between NFAT protein expression levels and various characteristics showed a significant association with histology (p=0.008, OR=0.273). CONCLUSION: Our results revealed that high NFAT expression was present in Chinese NSCLCs and that NFAT expression might be involved in the process of human lung cancer development. PMID- 17922468 TI - Localization and activity of iNOS in normal human lung tissue and lung cancer tissue. AB - Inducible nitric oxide synthase (iNOS) is one of three enzymes generating nitric oxide (NO) from the amino acid L-arginine. iNOS-derived NO plays an important role in several physiological and pathophysiological conditions. NO is a free radical which produces many reactive intermediates that account for its bioactivity. In the human lung, the alveolar macrophage is an important producer of cytokines and this production may be modified by NO. Moreover, high concentrations of NO have been shown to increase nuclear factor kappaB (NF-kB) activation. Recent investigations of NO expression in tumor tissue indicated that, at least for certain tumors, NO may mediate one or more roles during the growth of human cancer. We have studied iNOS in two tissue groups: normal human lung tissue and human lung cancer tissue. We localized iNOS in these tissues by immunohistochemistry and tested the mRNA expression by RT-PCR, the protein level by Western blot, and the protein activity by radiometric analysis. The results demonstrate different expression, localization and activity of iNOS in normal versus tumor tissue. This is suggestive of a role for NO production from iNOS in human lung cancer because high concentrations of this short molecule may transform to highly reactive compounds such as peroxynitrite (ONOO-); moreover, through the upregulator NF-kB, they can induce a chronic inflammatory state representing an elevated risk for cell transformation to cancer. PMID- 17922470 TI - Abstracts from the 4th Symposium on Controversies in Dialysis Access, October 18 19, 2007, Boston, Massachusetts, USA. PMID- 17922471 TI - Angioaccess in hemodialysis. Abstracts of the 5th International Congress of the Vascular Access Society, June 11-13 - 2007, Nice, France. PMID- 17922469 TI - Confidence interval for DNA/mRNA concentration by real-time PCR. PMID- 17922472 TI - PAI-1 activity, but not fibrinogen or von Willebrand factor, is inversely related to LDL particle size in type 2 diabetes. AB - BACKGROUND: Levels of fibrinogen, von Willebrand factor (vWF) and plasminogen activator inhibitor-1 (PAI-1) have been associated with small low-density lipoprotein (LDL) particles. However, it is not clear whether these associations are independent of visceral adiposity or other components of the metabolic syndrome such as triglycerides or insulin resistance. METHODS: Visceral adipose tissue (VAT; CT-scan), fibrinogen, von Willebrand factor antigen (vWF:Ag), PAI-1 activity and different metabolic parameters such as total cholesterol (chol), HDL chol, triglycerides, insulin resistance (homeostasis model assessment; HOMA-IR) were determined in 41 women and 78 men with type 2 diabetes. LDL particle size was assessed by polyacrylamide gradient gel electrophoresis. RESULTS: PAI-1 activity was inversely related to LDL particle size after adjustment for age and body mass index (BMI) (r=-0.28; p=0.006) or age and VAT (r=-0.26; p=0.01), but not after adjustment for age and HOMA-IR (r=-0.15; p=0.148) or age and triglycerides (r=-0.04; p=0.679). In multiple regression analysis, LDL particle size did not independently determine PAI-1 activity levels. Fibrinogen and vWF:Ag did not seem to be related to LDL size. CONCLUSIONS: PAI-1 activity levels, in contrast to fibrinogen and vWF:Ag, seem to be related to the small LDL phenotype in patients with type 2 diabetes. However, this relationship was not independent of insulin resistance or triglycerides. PMID- 17922473 TI - Genetic study of Saudi diabetes (GSSD): significant association of the KCNJ11 E23K polymorphism with type 2 diabetes. AB - BACKGROUND: The E23K variant of KCNJ11 has been associated with type 2 diabetes (T2D) in several but not all populations studied. Thus far, despite a high incidence of T2D, the role of this variant in Arabs has not been established. METHODS: We performed a case-control association study using 550 T2D Saudi patients (WHO criteria), and 335 controls (age>or=60; fasting plasma glucose<7 mmol/L). E23K genotyping was performed by using molecular beacon-based real time PCR assays. RESULTS: The difference in K or risk allele frequency of cases and controls was significant with an OR of 1.7 (p=0.0001). The K allele is more common among T2D patients (21%) than in the age and sex matched controls (13.6%). This was consistent with a likely eventual conversion to T2D of younger normoglycemic individuals as they grow older. CONCLUSIONS: Our results report for the first time a positive association of the E23K variant with T2D in an Arab population. Confirmation by a larger study is indicated. PMID- 17922474 TI - High-resolution analysis of DNA copy number alterations and gene expression in renal clear cell carcinoma. AB - We analysed chromosomal copy number aberrations (CNAs) in renal cell carcinomas by array-based comparative genomic hybridization, using a genome-wide scanning array with 2304 BAC and PAC clones covering the whole human genome at a resolution of roughly 1.3 Mb. A total of 30 samples of renal cell carcinoma were analysed, including 26 cases of clear cell carcinoma (CCC) and four cases of chromophobe renal cell carcinoma (ChCC). In CCCs, gains of chromosomes 5q33.1 qter (58%), 7q11.22-q35 (35%) and 16p12.3-p13.12 (19%), and losses of chromosomes 3p25.1-p25.3 (77%), 3p21.31-p22.3 (81%), 3p14.1-p14.2 (77%), 8p23.3 (31%), 9q21.13-qter (19%) and 14q32.32-qter (38%) were detected. On the other hand, the patterns of CNAs differed markedly between CCCs and ChCCs. Next, we examined the correlation of CNAs with expression profiles in the same tumour samples in 22/26 cases of CCC, using oligonucleotide microarray. We extracted genes that were differentially expressed between cases with and without CNAs, and found that significantly more up-regulated genes were localized on chromosomes 5 and 7, where recurrent genomic gains have been detected. Conversely, significantly more down-regulated genes were localized on chromosomes 14 and 3, where recurrent genomic losses have been detected. These results revealed that CNAs were correlated with deregulation of gene expression in CCCs. Furthermore, we compared the patterns of genomic imbalance with histopathological features, and found that loss of 14q appeared to be a specific and additional genetic abnormality in high grade CCC. When we compared the expression profiles of low-grade CCCs with those of high-grade CCCs, differentially down-regulated genes tended to be localized on chromosomes 14 and 9. Thus, it is suggested that copy number loss at 14q in high grade CCC may be involved in the down-regulation of genes located in this region. PMID- 17922477 TI - Size and distance. PMID- 17922476 TI - Insulin glargine and receptor-mediated signalling: clinical implications in treating type 2 diabetes. AB - Most patients with type 2 diabetes mellitus will eventually require insulin therapy to achieve or maintain adequate glycaemic control. The introduction of insulin analogues, with pharmacokinetics that more closely mimic endogenous insulin secretion, has made physiologic insulin replacement easier to achieve for many patients. However, there are also concerns regarding alteration of binding affinities for the insulin receptor (IR) or insulin-like growth factor-1 receptor (IGF-1R) may increase the mitogenic potential of some analogues. Therefore, this article will review the relevant preclinical and clinical data to assess the mitogenic potential of insulin glargine, a basal insulin analogue, compared with regular human insulin (RHI). Searches of the PubMed database were performed using terms that included 'IR,' 'insulin-like growth factor-1,' 'IGF-1R,' 'type 2 diabetes mellitus,' and 'insulin glargine.' Original articles and reviews of published literature were retrieved and reviewed. Although one study reported increased binding affinity of insulin glargine for the IGF-1R and increased mitogenic potential in cells with excess IGF-1Rs (Saos/B10 osteosarcoma cells), most in vitro binding-affinity and cell-culture studies have demonstrated behaviour of insulin glargine comparable to that of RHI for both IR and IGF-1R binding, insulin signalling, and metabolic and mitogenic potential.Currently published in vivo carcinogenic studies and human clinical trial data have shown that insulin glargine is not associated with increased risk for either cancer or the development or progression of diabetic retinopathy. PMID- 17922475 TI - Rosiglitazone increases matrix production and quenches inflammation: studies in human cells. AB - BACKGROUND: Type 2 diabetes (T2D) is characterized by an accelerated atherogenesis, a process to which both proliferative and inflammatory responses contribute. Peroxisome proliferator-activated receptors-gamma (PPARgamma) agonists have both anti-proliferative and anti-inflammatory properties. We tested the effect of therapeutic doses of rosiglitazone on proliferative and inflammatory pathways in fibroblasts (HF) from five controls (C) and five T2D patients, and in aortic smooth muscle cells (hSMC). METHODS: Transforming growth factor-beta (TGFbeta) and interleukin-6 (IL-6) expression, and IL-6, laminin and fibronectin release were measured. To identify the involved intracellular signalling, extracellular signal-regulated kinases (ERK)1/2 phosphorylation and p38 activation were evaluated. RESULTS: Both phorbol 12-myristate 13-acetate (PMA) [a protein kinase C (PKC) activator] and rosiglitazone increased TGFbeta expression and fibronectin and laminin release in C and T2D patients. Rosiglitazone effect was reversed by its specific inhibitor Sr202. The combination PMA + rosiglitazone was additive in C, but not in T2D patients. IL-6 production was stimulated by PMA in both C and T2D patients; this effect was prevented by rosiglitazone in a Sr202-inhibitable manner. Experiments performed in hSMC yielded the same results. Rosiglitazone increased p38 activation more in C than in T2D patients; PMA-induced phosphorylation of ERK1/2 was similarly reduced in both cells. CONCLUSIONS: In HF and hSMC, rosiglitazone stimulates the synthesis of matrix components via enhanced TGFbeta expression; when combined with PMA, the resulting PKC activation is mediated by enhanced p38 phosphorylation. On the other hand, rosiglitazone quenches inflammation in both cell types, by counteracting PMA-induced phosphorylation of ERK1/2. PMID- 17922478 TI - Motivational interviewing in musculoskeletal care. AB - Motivational interviewing is a patient-centred counselling method designed to build motivation for behaviour change by resolving ambivalence. It was developed in the addictions field and has since been applied to medical and health promotion settings. This paper will provide a brief overview of the method and will discuss how it may be used in consultations for patients with musculoskeletal problems to increase engagement with treatment and to build motivation for helpful behaviour change. PMID- 17922480 TI - Pleiotropy and principal components of heritability combine to increase power for association analysis. AB - When many correlated traits are measured the potential exists to discover the coordinated control of these traits via genotyped polymorphisms. A common statistical approach to this problem involves assessing the relationship between each phenotype and each single nucleotide polymorphism (SNP) individually (PHN); and taking a Bonferroni correction for the effective number of independent tests conducted. Alternatively, one can apply a dimension reduction technique, such as estimation of principal components, and test for an association with the principal components of the phenotypes (PCP) rather than the individual phenotypes. Building on the work of Lange and colleagues we develop an alternative method based on the principal component of heritability (PCH). For each SNP the PCH approach reduces the phenotypes to a single trait that has a higher heritability than any other linear combination of the phenotypes. As a result, the association between a SNP and derived trait is often easier to detect than an association with any of the individual phenotypes or the PCP. When applied to unrelated subjects, PCH has a drawback. For each SNP it is necessary to estimate the vector of loadings that maximize the heritability over all phenotypes. We develop a method of iterated sample splitting that uses one portion of the data for training and the remainder for testing. This cross validation approach maintains the type I error control and yet utilizes the data efficiently, resulting in a powerful test for association. PMID- 17922479 TI - Understanding the accuracy of statistical haplotype inference with sequence data of known phase. AB - Statistical methods for haplotype inference from multi-site genotypes of unrelated individuals have important application in association studies and population genetics. Understanding the factors that affect the accuracy of this inference is important, but their assessment has been restricted by the limited availability of biological data with known phase. We created hybrid cell lines monosomic for human chromosome 19 and produced single-chromosome complete sequences of a 48 kb genomic region in 39 individuals of African American (AA) and European American (EA) origin. We employ these phase-known genotypes and coalescent simulations to assess the accuracy of statistical haplotype reconstruction by several algorithms. Accuracy of phase inference was considerably low in our biological data even for regions as short as 25-50 kb, suggesting that caution is needed when analyzing reconstructed haplotypes. Moreover, the reliability of estimated confidence in phase inference is not high enough to allow for a reliable incorporation of site-specific uncertainty information in subsequent analyses. We show that, in samples of certain mixed ancestry (AA and EA populations), the most accurate haplotypes are probably obtained when increasing sample size by considering the largest, pooled sample, despite the hypothetical problems associated with pooling across those heterogeneous samples. Strategies to improve confidence in reconstructed haplotypes, and realistic alternatives to the analysis of inferred haplotypes, are discussed. PMID- 17922481 TI - A genomic imprinting test for ordinal traits in pedigree data. AB - Genomic imprinting can lead maternally and paternally derived alleles with identical nucleotide sequences to function differently and has been found to affect the complex inheritance of a variety of human disorders. Statistical methods that differentiate the parent-of-origin effects on human diseases are available for binary traits and continuous traits. However, numerous common diseases are measured on discrete ordinal scales. Imprinting may also contribute to the complex genetic basis of these traits. In a previous study, we proposed a latent variable model and developed computationally efficient score statistic to test linkage of ordinal traits for any size pedigree while adjusting for non genetic covariates. In this study, we extend the latent variable model to incorporate parent-of-origin information and further develop a score statistic for testing the imprinting effect in linkage analysis. We evaluated the properties of our test statistic using simulations. We then applied our method to the Collaborative Study on the Genetics of Alcoholism and found a novel locus on chromosome 18 that shows a strong signal for imprinting. In addition, we identified two loci on chromosomes 3 and 4 significantly (p<0.0001) linked with alcoholism. PMID- 17922482 TI - Novel application of flow cytometry: determination of muscle fiber types and protein levels in whole murine skeletal muscles and heart. AB - Conventional methods for measuring proteins within muscle samples such as immunohistochemistry and western blot analysis can be time consuming, labor intensive and subject to sampling errors. We have developed flow cytometry techniques to detect proteins in whole murine heart and skeletal muscle. Flow cytometry and immunohistochemistry were performed on quadriceps and soleus muscles from male C57BL/6J, BALB/c, CBA and mdx mice. Proteins including actins, myosins, tropomyosin and alpha-actinin were detected via single staining flow cytometric analysis. This correlated with immunohistochemistry using the same antibodies. Muscle fiber types could be determined by dual labeled flow cytometry for skeletal muscle actin and different myosins. This showed similar results to immunohistochemistry for I, IIA and IIB myosins. Flow cytometry of heart samples from C57BL/6J and BALB/c mice dual labeled with cardiac and skeletal muscle actin antibodies demonstrated the known increase in skeletal actin protein in BALB/c hearts. The membrane-associated proteins alpha-sarcoglycan and dystrophin could be detected in C57BL/6J mice, but were decreased or absent in mdx mice. With the ability to label whole muscle samples simultaneously with multiple antibodies, flow cytometry may have advantages over conventional methods for certain applications, including assessing the efficacy of potential therapies for muscle diseases. PMID- 17922483 TI - A possible link between the pubertal growth of girls and breast cancer in their daughters. AB - One hypothesis for the origins of breast cancer is that it is initiated by exposure of developing breast tissue in utero to maternal sex hormones. The sex hormone profile is established at puberty, when it regulates growth of the pelvic bones. The pubertal growth of girls is characterized by broadening and rounding of the pelvis. The maximal width between their iliac crests, the intercristal width, increases more rapidly than in boys. We hypothesized that higher sex hormone concentrations at puberty produce larger intercristal widths, and these are markers of increased breast cancer risk in the next generation. We followed up 6,370 women who were born in Helsinki during 1934-1944, and whose mothers' pelvic bones were measured during routine antenatal care. Women whose mothers had large intercristal widths had higher rates of breast cancer. In those born at or after 40 weeks gestation, the hazard ratio for breast cancer was 3.7 (95% CI: 2.1 6.6) if their mother's intercristal width was greater than 30 cm. Among women born to multiparous mothers this hazard ratio rose to 7.2 (3.4-15.4). Hazard ratios for breast cancer were also higher in the daughters of mothers with round iliac crests. Pelvic bone measurements which increase similarly in girls and boys at puberty did not predict breast cancer. We conclude that the intercristal width, and the roundness of the iliac crests, are markers of mothers' sex hormones, and postulate that high concentrations cause genetic instability in differentiating breast cells in their daughters in utero. PMID- 17922484 TI - Prompt disulfide fragmentations of disulfide-containing proteins in a matrix assisted laser desorption/ionization source. PMID- 17922485 TI - Mass spectrometry characterization of Escherichia coli K4 oligosaccharides from 2 mers to more than 20-mers. AB - The separation and characterization of oligosaccharides obtained by hyaluronidase [E.C. 3.2.1.35] digestion of Escherichia coli K4 polysaccharide using online high performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS) are presented. Complete identification and structural information for oligosaccharides containing 2-24 monomers (from 2- to 24-mers) were obtained. In particular, smaller K4 species, from 2-mers to 4-mers, exhibited mainly [M-H]( 1) anions, whereas the 6- to 8-mers existed predominantly at the charge state of 2. The K4 oligomers from 10-mers to 14-mers were mainly represented by [M-3H](-3) anions while species from 16- to 20-mers were characterized by a charge state of 4. K4 oligosaccharides from 22- to 24-mers existed as [M-4H](-4) and [M-5H](-5) anions and, for this latter species, ions having a charge state of -6 appeared. For smaller K4 species, in particular from 6-mers to 10-mers, ESI-MS revealed anions related to the loss of one monosaccharide unit from the oligomers due to apparent collisional activation and ion source fragmentation. However, no odd numbered anions were produced for K4 2/4-mer species or for oligosaccharides greater than 12-mers, while for K4 species 8/10-mer, ESI-MS revealed odd-numbered anions generally in low relative abundance making the interpretation of the spectra easier. The ESI-MS spectra of oligosaccharides separated by online HPLC were applied to the evaluation of the K4 polymerization process, confirming that the addition of fructose units is not critical for chain elongation as variously fructosylated oligomer species were detected directly on the K4 carbohydrate backbone. PMID- 17922486 TI - A generic fast solid-phase extraction high-performance liquid chromatography/mass spectrometry method for high-throughput drug discovery. AB - High-performance liquid chromatography/mass spectrometry (HPLC/MS) is increasingly perceived to be an essential tool in drug discovery at many key steps, like drug screening, lead identification, ADME profiling, and drug metabolism and pharmacology studies. High-throughput screenings in the early phase for metabolic stability, protein binding, permeability (ADME) and bioavailability are widely used to weed out compounds that do not exhibit the necessary characteristics. For such high-throughput LC/MS studies, a generic LC/MS method that can be used for a variety of compounds is desired. In this study, we used a small set of compounds with a wide range of properties to guide method development, and achieved a sample throughput of 1.7 min/sample. Here, we present a generic fast method that achieves good peak separation and peak shape on conventional HPLC systems using a column-switching mechanism for on-line solid phase extraction (SPE)-HPLC/MS analysis. The method has a linear response range from 1 to 500 nM for the tested compounds. When a larger set of 658 randomly picked small molecules were analyzed using this method, 612 were observed with good signal intensity and HPLC peak shapes. This generic fast SPE-LC/MS method has been used to screen more than 1.5 million compounds repetitively against over 200 protein targets for hit confirmation and semi-quantitation of binding constants from biological assays. Over 7000 different compounds for a variety of protein-binding assays have been studied using this method for quantitative analysis as well. PMID- 17922487 TI - Differentiation of diastereomeric N aryltetrahydropyrano/tetrahydrofuranochromenylamines under electron ionization and chemical ionization conditions. AB - A series of diastereomeric 4S,5S,6R/S-tetrahydropyrano- and 3S,4S,5R/S tetrahydrofuranochromenylamine derivatives (a/b isomers; 1-26) has been studied under electron ionization (EI) and chemical ionization (CI) conditions. The EI mass spectra of all diastereomeric compounds show two characteristic fragment ions, of which one is formed by retro-Diels-Alder (RDA) reaction from the molecular ion, retaining the charge on the diene fragment, and the other [M (HNAr)]+ ion by a simple radical loss. The RDA process is more favorable in all b isomers, whereas the radical loss is dominant in all a isomers; based on these two ions it is easy to differentiate the two diastereomers. The collision-induced dissociation (CID) spectra of all the molecular ions also show the same trend, which reflects the stereoselectivity in the formation of the two characteristic fragment ions. The results of theoretical calculations performed are in accordance with the experimental observations. The CI experiments (methane and isobutane) on all the diastereomeric compounds also enabled the differentiation of the isomers. PMID- 17922488 TI - Quantitative determination of lysozyme-ligand binding in the solution and gas phases by electrospray ionisation mass spectrometry. AB - Affinity constants for the binding of a range of substrate and non-substrate oligosaccharides to hen egg white lysozyme were determined by direct observation of the protein.ligand complexes using electrospray ionisation mass spectrometry (ESI-MS) with a chip-based nano-ESI source. The values obtained for a series of beta-1,4-N-acetylglucosamine oligomers (NAGn) were found to be in good agreement with those determined by fluorescence measurement. Oligomers of alpha-1,4-glucose (Glcn), which are believed to bind to lysozyme non-specifically, exhibited a 10(6)- to 10(8)-fold lower affinity for the enzyme. Lysozyme.NAGn complexes displayed an increase in Ka from n=2 to n=4, but then reached a plateau. In contrast non-specific lysozyme.Glcn complexes showed no such trend. Determination of gas-phase complex stability was achieved by quantitative collision-induced dissociation (CID) and infrared multiphoton dissociation (IRMPD) measurements. The collision energy (Ec50) or laser power (IRMPD50) required to dissociate precursor ions to 50% of their original intensity was determined for lysozyme.NAGn and Glcn complexes using the [M+8H]8+ charge state. An excellent correlation between trends in Ka and gas-phase stability was seen for NAGn oligomers bound to lysozyme, whereas no such relationship was observed with the non-specific, weaker lysozyme.Glcn complexes. These results illustrate that ESI MS can be used to quantify the interactions between lysozyme and oligosaccharides in both the solution and gas phase and that measurement of gas-phase complex stability by CID or IRMPD can provide information about specific solution binding events. PMID- 17922489 TI - Multi-class confirmatory method for analyzing trace levels of tetracyline and quinolone antibiotics in pig tissues by ultra-performance liquid chromatography coupled with tandem mass spectrometry. AB - An ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC/MS/MS) method was developed to screen and confirm multi-class veterinary drug residues in pig tissues including pig kidney, liver and meat. Twenty-one drugs of two different classes including seven tetracyclines and four types of quinolones (quinoline, naphthyridine, pyridopyrimidine and cinoline) were determined simultaneously in a single run. The homogenized sample tissues were extracted with EDTA-McIlvaine buffer solution and further purified using a polymer-based Oasis HLB solid-phase extraction (SPE) cartridge. An ACQUITY UPLC BEH C18 column was used to separate the analytes followed by tandem mass spectrometry using an electrospray ionization source. MS data acquisition was performed in the positive ion multiple reaction monitoring mode, selecting two ion transitions for each target compound. Recovery studies were performed at different fortification levels. The overall average recoveries from pig muscle, kidney, and liver fortified with quinolones and tetracyclines at three levels ranged from 80.2 to 117.8% based on the use of matrix-fortified calibration with the coefficients of variation ranging from 2.1 to 17.8% (n=6). The limits of quantitation (LOQs) of quinolones and tetracyclines in different tissues ranged from 0.03-4.50 microg/kg and 0.16-10.00 microg/kg, respectively. The effects of the extraction solvent, SPE cartridge, elution solvent and sample matrix on the analyte recovery as well as the effects of the mobile phase composition and column temperature on the chromatographic behavior were also studied. PMID- 17922490 TI - Nursing support at the onset of rheumatoid arthritis: Time and space for emotions, practicalities and self-management. AB - BACKGROUND: Following a diagnosis of rheumatoid arthritis (RA), patients have to adapt to lifelong, unpredictable but repeated episodes of pain and disability, potentially leading to permanent loss of function and its consequences on their lives. We established nurse clinics with the aim of supporting newly diagnosed RA patients in adapting to and managing their long-term condition. The aim of this study was to explore the content of clinic discussions in this new clinical service, in order to ascertain patients' needs upon diagnosis. METHODS: All clinic letters from the nurse to the family doctor were analysed. Every topic mentioned was systematically coded independently by a researcher and a patient research partner, who compared and agreed codes. Codes were organized into categories, and, finally, into overarching themes. RESULTS: Twenty-four patients had 74 appointment letters. A total of 79 codes were identified, from which ten categories emerged, and, finally, three overarching themes. The first theme related to 'Emotional support', which underpinned the other two themes and was discussed in almost all appointments. Issues included discussions about the emotional consequences of RA, needing time to adjust, frustration and fears for the future. The second theme 'Practicalities of the treatment of RA', included subordinate themes relating to the nature of RA, such as identity, cause, timeline, consequences and treatment. Medication issues were discussed and referrals to the multidisciplinary team were made. The final theme related to the 'Self-management of RA', and included discussions on physical symptoms and their management. CONCLUSIONS: The offer to attend a nurse clinic soon after diagnosis allowed RA patients to discuss a wide range of practical and self-management issues. However, most patients also took the opportunity and time to discuss emotional reactions and adaptations to diagnosis. The data suggest an unmet need for emotional support that a nurse clinic might be able to provide. PMID- 17922491 TI - A comparison of methods for combining quality and efficiency performance measures: profiling the value of hospital care following acute myocardial infarction. AB - Health plans have begun to combine data on the quality and cost of medical providers in an attempt to identify and reward those that offer the greatest 'value.' The analytical methods used to combine these measures in the context of provider profiling have not been rigorously studied. We propose three methods to measure and compare the value of hospital care following acute myocardial infarction by combining a single measure of quality, in-hospital survival, and the cost of an episode of acute care. To illustrate these methods, we use administrative data for heart attack patients treated at 69 acute care hospitals in Massachusetts in fiscal year 2003. In the first method we reproduce a common approach to value profiling by modeling the two case mix-standardized outcomes independently. In the second approach, survival is regressed on patient risk factors and the average cost of care at each hospital. The third method models survival and cost for each hospital jointly and combines the outcomes on a common scale using a cost-effectiveness framework. For each method we use the resulting parameter estimates or functions of the estimates to compute posterior tail probabilities, representing the probability of being classified in the upper or lower quartile of the statewide distribution. Hospitals estimated to have the highest and lowest value according to each method are compared for consistency, and the advantages and disadvantages of each approach are discussed. PMID- 17922492 TI - A FAK/Src chimera with gain-of-function properties promotes formation of large peripheral adhesions associated with dynamic actin assembly. AB - Formation of a complex between the tyrosine kinases FAK and Src is a key integrin mediated signaling event implicated in cell motility, survival, and proliferation. Past studies indicate that FAK functions in the complex primarily as a "scaffold," acting to recruit and activate Src within cell/matrix adhesions. To study the cellular impact of FAK-associated Src signaling we developed a novel gain-of-function approach that involves expressing a chimeric protein with the FAK kinase domain replaced by the Src kinase domain. This FAK/Src chimera is subject to adhesion-dependent activation and promotes tyrosine phosphorylation of p130Cas and paxillin to higher steady-state levels than is achieved by wild-type FAK. When expressed in FAK -/- mouse embryo fibroblasts, the FAK/Src chimera resulted in a striking cellular phenotype characterized by unusual large peripheral adhesions, enhanced adhesive strength, and greatly reduced motility. Live cell imaging of the chimera-expressing FAK -/- cells provided evidence that the large peripheral adhesions are associated with a dynamic actin assembly process that is sensitive to a Src-selective inhibitor. These findings suggest that FAK-associated Src kinase activity has the capacity to promote adhesion integrity and actin assembly. PMID- 17922493 TI - Portuguese version of the Delirium Rating Scale-Revised-98: reliability and validity. AB - OBJECTIVE: To assess the validity and the reliability of the Portuguese version of the Delirium Rating Scale-Revised-98 (DRS-R-98). METHODS: The scale was translated into Portuguese and back-translated into English. After assessing its face validity, five diagnostic groups (n=64; delirium, depression, dementia, schizophrenia and others) were evaluated by two independent researchers blinded to the diagnosis. Diagnosis and severity of delirium as measured by the DRS-R-98 were compared to clinical diagnosis, Mini-Mental State Exam, Confusion Assessment Method, and Clinical Global Impressions scale (CGI). RESULTS: Mean and median DRS R-98 total scores significantly distinguished delirium from the other groups (p<0.001). Inter-rater reliability (ICC between 0.9 and 1) and internal consistency (alpha=0.91) were very high. DRS-R-98 severity scores correlated highly with the CGI. Mean DRS-R-98 severity scores during delirium differed significantly (p<0.01) from the post-treatment values. The area under the curve established by ROC analysis was 0.99 and using the cut-off value of 20 the scale showed sensitivity and specificity of 92.6% and 94.6%, respectively. CONCLUSION: The Portuguese version of the DRS-R-98 is a valid and reliable measure of delirium that distinguishes delirium from other disorders and is sensitive to change in delirium severity, which may be of great value for longitudinal studies. PMID- 17922494 TI - Simple machine-assisted protocol for solid-phase synthesis of depsipeptides. AB - A straightforward machine-assisted protocol for the synthesis of linear depsipeptides is reported. The synthesis was performed on a 433A Peptide Synthesizer (Applied Biosystems) using preprogrammed and optimized modules for Boc chemistry and without any need for hardware modification. The robustness of the protocol was demonstrated with 12 examples of 26-membered depsipeptides with single and multiple (up to 6) ester backbone substitutions. PMID- 17922495 TI - Secondary structure inducing potential of beta-amino acids: torsion angle clustering facilitates comparison and analysis of the conformation during MD trajectories. AB - While numerous examples of beta-peptides--exclusively composed of beta-amino acids--have been investigated during the past decade, there are only few reports on the conformational preference of a single beta-amino acid when incorporated into a cyclopeptide. The conformational bias of beta-amino acids on the secondary structure of cyclopeptides has been investigated by NMR spectroscopy in combination with distance geometry (DG) and molecular dynamics (MD) calculations using experimental constraints. The atomic coordinate RMSD criterion usually employed for clustering of conformations after DG and MD calculations does not necessarily group similar peptide conformations, as there is an insufficient correlation between atomic coordinates and torsion angles. To improve on this shortcoming and to eliminate any arbitrary decisions during this process, a torsion angle clustering procedure has been implemented. For the cyclic pentapeptides cyclo-(-Val-beta-Hala-Phe-Leu-Ile-) 1 and cyclo-(-Ser-Pro-Leu-beta Hasn-Asp-) 3, the beta-amino acid is found in the central position of an extended gamma-turn (pseudo gamma-turn, Psigamma-turn), while the beta-Hpro residue in the cyclic hexapeptide cyclo-(-Ser-beta-Hpro-Leu-Asn-Ile-Asp-) 5 preferentially occupies position i+1 of a pseudo beta-turn (Psibeta-turn). These results further corroborate the hypothesis of beta-amino acids being reliable inducers of secondary structure in cyclic penta- and hexapeptides. They can be employed in the de novo design of biologically active cyclopeptides in pharmaceutical research, since the three-dimensional presentation of pharmacophoric groups in the side chains can be tailored by incorporation of beta-amino acids in strategic sequential positions. PMID- 17922496 TI - Copper binding and conformation of the N-terminal octarepeats of the prion protein in the presence of DPC micelles as membrane mimetic. AB - The prion protein is usually pictured as globular structured C-terminal domain that is linked to an extended flexible N-terminal tail. However, in its physiological form, it is a glycoprotein tethered to the cell surface via a C terminal GPI anchor. The low solubility of PrP even without GPI anchor and its strong tendency for aggregation has forced most structural investigations to be performed at low pH and mostly with N-terminally truncated variants. In the present study, we have used a synthetic peptide related to the PrP tetra octarepeat region, i.e., the sequence (Pro-His-Gly-Gly-Gly-Trp-Gly-Gln)(4), for NMR structural analysis of its preferred conformation in DPC micelles as membrane mimic. Well-defined and identical loops are observed between the four octarepeats that are linked by flexible Gly-Gly-Gly sequences. Interaction with the micelles is mainly through the tryptophan residues that appear to act as anchors. Copper binding to the peptide in the presence of DPC micelles revealed marked conformational rearrangements although binding to the micelles is preserved. Interestingly, titration experiments point to cooperative effects for the four binding sites. A destabilization of the DPC micelles by the peptide parallels the destabilizing effect of the prion protein on membranes so that the octarepeat region appears to be very membrane-active. How the physico-chemical properties reported here are linked to the function and significance of the prion protein remains a puzzle as long as the functional mechanism of the prion protein is not precisely elucidated. Nevertheless, our results emphasize the strong influence of the (membrane) environment on the PrP properties. PMID- 17922497 TI - Remembering Bruce: the early years. AB - This paper summarizes Merrifield-Stewart interactions during the years 1952-1968. PMID- 17922498 TI - The PTB domain of tensin: NMR solution structure and phosphoinositides binding studies. AB - Tensin is a protein confined at those discrete and specialized regions of the plasma membrane, known as focal adhesions. It contains, at the C-terminus, a phosphotyrosine binding (PTB) domain that can interact with the cytoplasmic tail of beta-integrins and is necessary for localization of the protein to cell-matrix adhesions. Here, we present the NMR solution structure of the PTB domain of tensin1. Moreover, through NMR binding studies, we demonstrate that the PTB domain of tensin1 is able to interact with phosphatidylinositol 4, 5-diphosphate (PtIns(4,5)P2) and phosphatidylinositol 4-phosphate (PtIns(4)P), presenting higher affinity for the diphosphorylated inositide. Chemical shift mapping studies reveal a putative PtIns(4,5)P2 binding region that is distinct from the predicted integrin beta-tail recognition site. Heteronuclear NOE experiments, recorded in absence and presence of PtIns(4,5)P2, indicate that the interaction with lipids decreases the flexibility of loop regions, predicted to be important for integrin binding, and thus, proposes a possible correlation between the two distinct binding events. Therefore, our studies suggest that capture of lipids by the PTB domain of tensin1 may play a role for the protein function at focal adhesions. PMID- 17922499 TI - Enantioseparation of phenothiazines in CD-modified CZE using single isomer sulfated CD as a chiral selector. AB - Enantioseparations of five chiral phenothiazines in CD-modified CZE using the single isomer sulfate-substituted beta-CD (heptakis(2,3-dihydroxy-6-O-sulfo)-beta CD, SI-S-beta-CD) and dual CD systems consisting of SI-S-beta-CD and a neutral CD as chiral selectors in a citrate buffer at pH 3.0 were investigated. The results indicate that SI-S-beta-CD is an excellent chiral selector for enantioseparation of promethazine. The enantiomers of trimeprazine were well separated, while those of ethopropazine could also be baseline-resolved with SI-S-beta-CD. With dual CD systems, especially with hydroxypropyl-beta-CD (HP-beta-CD) as neutral CD, the enantioselectivity of thioridazine and ethopropazine was considerably enhanced. Effective enantioseparation of phenothiazines, except for methotrimeprazine, could thus be favorably and simultaneously achieved. Moreover, reversal of the enantiomer migration order of ethopropazine and thioridazine occurred by varying the concentration of gamma-CD in the presence of SI-S-beta-CD. These phenomena may be attributable to the opposite effects of sulfated beta-CD and gamma-CD on the mobility of the enantiomers of ethopropazine and of thioridazine. Comparative studies on the enantioseparations of phenothiazines with single CD and dual CD systems containing SI-S-beta-CD and randomly sulfate-substituted beta-CD (MI-S beta-CD) were made. PMID- 17922500 TI - CE-based noncompetitive immunoassay for immunoglobulin G in bovine colostrum products. AB - A CE-based noncompetitive immunoassay for IgG in bovine colostrum products was established. FITC-labeled protein G (FITC-PrG) was tagged through noncovalent bindings to the Fc region of the mouse monoclonal antibovine IgG (Ab). The FITC PrG, Ab, and IgG formed a sandwiched immunocomplex FITC-PrG-Ab-IgG under optimal incubation conditions. The immunocomplex was separated and analyzed by CZE with LIF detection in less than 2 min in an uncoated fused-silica capillary. Addition of PEG 20,000 (PEG 20M) in the running buffer significantly suppressed analyte adsorption and thus improved the reproducibility and the resolution. The precision of the method was 5.1% (n = 7). A linear relationship was established for the IgG concentration in the range of 1-5 mg/L with a linear correlation coefficient (r = 0.9917). The LOD was 0.1 mg/L (S/N = 3). The method was successfully applied for the determination of IgG in bovine colostrum products and satisfactory results were achieved. PMID- 17922501 TI - High-throughput determination of glutathione and reactive oxygen species in single cells based on fluorescence images in a microchannel. AB - A novel high-throughput method is presented based on fluorescence images of cells in a microchannel for determination of glutathione (GSH) and reactive oxygen species (ROS) inside single cells. We first present a method to determine GSH and ROS separately, in which GSH in cells is derivatized by 2,3 naphthalenedicarboxaldehyde (NDA), and intracellular ROS is labeled using dihydrorhodamine 123. The cells with either fluorescent derivatized GSH or fluorescent labeled ROS are introduced into a microchannel and fluorescence images of every moving cell in the microchannel are taken continuously using a highly sensitive thermoelectrically cooled electron-multiplying CCD. The fluorescence intensities of the images correspond to the masses of GSH or ROS. An average detection rate of 80-120 cells/min is achieved. We then propose a method for simultaneously determining GSH and ROS, in which ROS is first labeled in the cells. The labeled cells are then introduced into the whole channel and allowed to immobilize onto the glass substrate. The fluorescence images of all the cells in the channel are taken. NDA is then introduced into the channel to derivatize the GSH in the immobilized cells, and fluorescence images of all cells are taken again. An average analysis rate of 20 cells/min is achieved. The masses of GSH and ROS in the single cells can be obtained from the fluorescence intensities of the images using their calibration curves. Since the cells are not lysed, there is no problem with adsorption of biological macromolecules and cellular debris on the channel wall, so that channel treatment, necessary in usual single-cell analysis techniques using CE and microchip electrophoresis, is no longer necessary. For single global cells, this method can also be used to determine the concentrations of ROS and GSH, which has not been reported previously. The concentrations of ROS and GSH in single global cells can be calculated from the determined masses and the cell volume (derived from the diameter of the round fluorescence image of the derivatized GSH). For gastric cancer cells, the concentrations of GSH and ROS are in the range 0.35x10(-3)-1.3x10(-3) mol/L and 0.77x10(-) (6)-1.5x10(-6) mol/L, respectively. PMID- 17922502 TI - RNA analysis by MEKC with LIF detection. AB - We have developed and validated a procedure of high sensitivity for the analysis of RNA. The procedure is based on the separation and detection of the 5' monophosphates of ribonucleosides selectively conjugated with 4,4-difluoro-5,7 dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl ethylene diamine hydrochloride (BODIPY FL EDA) at the 5'-phosphate group using CE with LIF. BODIPY conjugates of the four common ribonucleoside-5'-monophosphates were prepared and subjected to CE-LIF to serve as standard compounds for peak assignment and to develop separation conditions. After digestion of RNA or oligoribonucleotides to 5' monophosphates by nuclease P1 and fluorescence labeling BODIPY conjugates were detected and resolved by CE-LIF without further purification steps. Comparative CE-LIF analyses with DNA digested to deoxyribonucleoside-5'-monophosphates showed that the assay is equally efficient and sensitive for RNA analysis. Conditions to determine the modified ribonucleosides inosine, xanthosine, pseudouridine and 2' O-methyladenosine were also established. The limits of detection were in the range of 80-200 pM. After calibrating the assay with oligoribonucleotides, pseudouridine was quantified in total RNA of Drosophila, human liver, human kidney and t-RNA of Saccharomyces cerevisiae. These studies demonstrate good potential of fluorescence labeling of ribonucleoside-5'-monophosphates with BODIPY FL EDA and detection by CE-LIF to determine RNA composition with high accuracy and sensitivity. PMID- 17922503 TI - Development of a CE method for the determination of mycophenolic acid in human plasma: a comparison with HPLC. AB - Mycophenolate mofetil (MMF) is a widely used drug for the maintenance of immunosuppressive therapy in renal-transplant recipients. MMF is rapidly metabolized in vivo to mycophenolic acid (MPA), a reversible, noncompetitive inhibitor of inosine monophosphate dehydrogenase, which represents a limiting enzyme in lymphocyte proliferation. MPA shows large interindividual pharmacokinetic variability: its monitoring is therefore of primary importance to achieve adequate immunosuppression with minimized risk of graft rejection or toxicity. We developed a CE method for the determination of total MPA (tMPA) in plasma, based on easy sample preparation; CE evaluation of tMPA was performed in 30 mmol/L sodium-borate with 10 mmol/L SDS (pH 10.00) at 25 degrees C using a 60 cm (54.5 to window) uncoated capillary with UV detection at 254 nm wavelength. MPA was readily detectable in plasma; the CE method was linear in the range of 0.7-120 microg/mL (r >0.992). Intra- and interassay imprecision was <7% except for the lowest spiked MPA concentration, which had an intra-assay RSD% of 14.7 compared to 18.3 interassay. Data by CE were compared with results obtained by a validated HPLC method. CE assay of tMPA exhibited a very good correlation (r(2) >0.988) with respect to HPLC; Bland-Altman difference versus average showed a mean of -0.18 microg/mL +/- 1.14 SD. CE determination of tMPA is a robust, sensitive and reproducible method with the advantage over HPLC of being fast, simple and unexpensive, also enabling quick assessment of MPA for pharmacokinetic studies. PMID- 17922504 TI - Analysis and purification of peptide nucleic acids by denaturing PAGE. AB - A flexible and convenient protocol for the analysis and purification of peptide nucleic acid (PNA) oligomers and PNA-peptide chimeras by denaturing PAGE is described. Vertical slab gel electrophoresis, 26% in polyacrylamide and 8 M urea at pH 3, was suitable for analysis of oligomers ranging in size from tetramers (4 mers) to tetradodecamers (24-mers). Single-base resolution of oligomers was achieved and separations are generally superior to those given by standard RP HPLC techniques. The separation of a related series of PNA oligomers showed the distance migrated was linearly dependent on the logarithm of the molecular weight. The migration of oligomers through the gel is dependent on the number of basic functional groups present, such as amino groups, and the A and C content of the oligomer. PNAs are amenable to detection by UV-shadowing technique illuminated at 260 nm or Coomassie blue staining, both with similar, sub microgram per band detection limits. PMID- 17922505 TI - Preparative alkaline urea gradient PAGE: application to purification of extraordinarily-stable disulfide-linked homodimer of human growth hormone. AB - The 40-60 pituitary human growth hormone (hGH) isoforms are so similar in their physico-chemical properties (charge, size, hydrophobicity) that the limited resolutions of chromatographic separation methodologies have not permitted most of them to be isolated. However, application of high-resolution preparative alkaline urea gradient PAGE has facilitated isolation of a disulfide-linked mercaptoethanol-resistant (MER) 45 kDa hGH dimer. Human pituitary extracts were separated by Sephadex G-100 chromatography under alkaline conditions. Pooled fractions containing MER-45 kDa hGH, as determined by SDS-PAGE, were then separated by Sephadex G-100 chromatography under acidic conditions followed by diethylaminoethyl (DEAE) anion-exchange chromatography. Pooled DEAE fractions containing MER-45 kDa hGH and other hGH isoforms were then separated by preparative electrophoresis in an alkaline polyacrylamide gradient (5-20%) slab gel containing 8 M urea into five distinct protein zones. One electroeluted zone contained pure MER-45 kDa hGH. The dimeric hGH isoform was immunoreactive at low concentrations (effective dose to produce 50% response (ED(50)) +/- S.E. = 58 +/- 5.00 pM) in a hGH radioimmunoassay, similar to that of standard monomeric hGH (ED(50) +/- S.E. = 22.93 +/- 3.90 pM), indicating that is was conformationally intact. Alkaline urea gradient PAGE is a valuable tool for preparative separation of structurally similar proteins such as isoforms of the hGH family. PMID- 17922506 TI - Carrier ampholytes for IEF, on their fortieth anniversary (1967-2007), brought to trial in court: the verdict. AB - The present review summarizes the data accumulated in 1-year work, by exploring, via a 3-D methodology (Rotofor fractionation followed by CE-MS), all the narrow (2 pH unit wide) carrier ampholyte (CA) compounds for IEF, produced by three companies (Pharmacia with Pharmalyte and Ampholine, BioRad with Bio-Lyte and Serva with Servalyt). All species have been assessed by measuring the types of pH gradient produced, the total number of individual chemicals (with M(r) values) and isoforms and their focusing behavior ('good' or 'poor' ampholytes). Servalyt contains a grand total of 686 chemical entities and no less than 3899 isoforms; Pharmalyte 643 and 2211; Bio-Lyte 255 and 1192; Ampholine 294 and 1182, respectively. In terms of M(r) distribution, although all 2-pH-unit ranges start with the same low M(r) values (ca. 200) their upper limits are quite different. Thus, Pharmalyte reaches an upper M(r) value of 1179 (in the pH 4-6 range), versus 907 for Servalyt, 835 for Bio-Lyte and 893 for Ampholine. In general, in going towards the more alkaline pH intervals (e.g. pH 8-10) the molecular mass of carrier ampholytes (CAs) is reduced to as low as 491 (Bio-Lyte), indicating that the alkaline species are probably made with shorter oligoamines and are, in general, less substituted. All acidic pH intervals (up to pH 6-8) appear to be constituted by a very large proportion of well focusing species, indicating small values of DeltapK across their pI. Above pH 8, all brands of CAs worsen, the vast majority being unable to focus properly and sustain adequately the pH gradient. General guidelines are given for the synthesis of new alkaline species for improving the basic pH ranges. PMID- 17922507 TI - Analysis of dideoxyadenosine triphosphate by CE with fluorescence detection. I. Derivatization through the phosphate group. AB - A CZE method was developed, which separates 2',3'-dideoxyadenosine-5' triphosphate (ddATP) from other metabolites and endogenous nucleotides at high concentrations (20-200 microg/mL) to allow UV detection. To enhance sensitivity, fluorescence detection which requires prior derivatization of compounds was examined. Precapillary derivatization of ddATP in the presence of N-(3 dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDAC) with dansyl ethylenediamine (dansyl EDA) was faster and stable compared to that of 4,4 difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl ethylenediamine (BODIPY FL EDA). Reaction conditions, reagent concentrations and detection parameters were optimized and highest derivatization efficiency was achieved in 0.1 M 1-methylimidazole buffer (pH 8.0) with 140 mM EDAC in 1-methylimidazole buffer and 30 mM dansyl EDA in DMF for 90 min at 60 degrees C. Dansyl EDA derivatives of ddATP, 2'-deoxyadenosine-5'-triphosphate (dATP) and ATP were comigrating with the CZE method; therefore, a MEKC method was developed and optimized for repeatable separations. Upon dansylation, sensitivity of ddATP with fluorescence detection (LOQ = 12 ng/mL) was 160 times higher than UV detection (LOQ = 1.9 microg/mL). PMID- 17922508 TI - A simple method to remove contaminating salt from IPG strips prior to IEF. AB - In this study, Saccharomyces cerevisiae yeast lysate is used to demonstrate how a simple wash procedure can improve IEF of IPG strips passively rehydrated in the presence of NaCl. By performing three 10 min washes after IPG strip rehydration and before IEF, corresponding second-dimensional gels from strips containing NaCl look similar to control strips while the second-dimensional gels of unwashed strips contains streaks and spaces devoid of protein. Up to 500 mM NaCl was added to the yeast lysate and successfully focused following this wash regime. Protein loss due to the washes was determined to be minimal by comparing replicates of washed and unwashed strips and analyzing the densities of their corresponding second-dimensional gel spots. In the event of unknown salt contamination, indicated by low voltage during focusing, it is possible to stop focusing, wash the strips, and then continue focusing with acceptable second dimension results. PMID- 17922509 TI - DNA is more negatively supercoiled in bacterial plasmids than in minichromosomes isolated from budding yeast. AB - A series of circular shuttle vectors were constructed that could replicate and transcribe in the cells of both Escherichia coli and Saccharomyces cerevisiae. 2 D agarose gel electrophoresis run without or in the presence of different concentrations of chloroquine (CHL) revealed that bacterial plasmids were more negatively (-) supercoiled than minichromosomes isolated from budding yeast. Attempts to increase (-) supercoiling in S. cerevisiae or to reduce it in E. coli have deleterious biological consequences. These observations indicate that DNA supercoiling can vary in different species but cells are exquisitely sensitive to sudden changes in supercoiling. In E. coli, the observation that cell growth as well as ColE1 plasmid copy number decrease when DNA relaxes suggests that supercoiling could affect cell viability by regulating the initiation of both transcription and replication. PMID- 17922510 TI - Using DNA sequencing electrophoresis compression artifacts as reporters of stable mRNA structures affecting gene expression. AB - The formation of secondary structure in oligonucleotide DNA is known to lead to "compression" artifacts in electropherograms produced through DNA sequencing. Separately, the formation of secondary structure in mRNA is known to suppress translation; in particular, when such structures form in a region covered by the ribosome either during, or shortly after, initiation of translation. Here, we demonstrate how a DNA sequencing compression artifact provides important clues to the location(s) of translation-suppressing secondary structural elements in mRNA. Our study involves an engineered version of a gene sourced from Rhodothermus marinus encoding an enzyme called Cel12A. We introduced this gene into Escherichia coli with the intention of overexpressing it, but found that it expressed extremely poorly. Intriguingly, the gene displayed a remarkable compression artifact during DNA sequencing electrophoresis. Selected "designer" silent mutations destroyed the artifact. They also simultaneously greatly enhanced the expression of the cel12A gene, presumably by destroying stable mRNA structures that otherwise suppress translation. We propose that this method of finding problem mRNA sequences is superior to software-based analyses, especially if combined with low-temperature CE. PMID- 17922511 TI - Deriphat 2-DE to visualize polyphenol oxidase in Moscato and Prosecco grapes. AB - The Deriphat 2-DE was used to visualize polyphenol oxidase (PPO) isoforms of Moscato and Prosecco grape extracts, partially purified and characterized. Catecholase has similar values in the two varieties, whereas Moscato cresolase data are almost 54% higher. In the first dimension, the PPO of both varieties may be detected by SDS-PAGE, but native PAGE (N-PAGE) gave negative results. For this reason, the samples were solubilized in the zwitteronic detergent Deriphat, which was also included in the gel and the cathodic buffer. Deriphat migrated together with the cathodic buffer, maintaining protein solubility and revealing the PPO profiles of Moscato and Prosecco extracts in native conditions. The combination of Deriphat-PAGE (D-PAGE) and SDS-PAGE (2-DE) also resulted in improved separation efficiency in resolving PPO and specialized stains in evaluating PPO activities. The control, represented by IEF for the first-dimensional separation, had a lower number of spots, demonstrating the higher capacity of Deriphat 2-DE to isolate PPO isoforms from grape extracts. The Deriphat 2-DE method described here is simple but powerful, and the resulting information will be a useful tool for further proteomic research. PMID- 17922512 TI - Partial characterization of the proteome of the mouse striatum. AB - Many diseases of the mammalian CNS, including Parkinson's (PD) and Lesch Nyhan disease (LND), are associated with programmatic neurodegeneration or dysfunction of dopaminergic neurons in the mesencephalon, the nigrostriatal pathway, and its projections in the striatum [1-4]. Proteomic studies on brain tissue of both animal models and human PD patients have provided evidence for dysfunction and damage of many pathways, including oxidative stress-related damage, ubiquitin proteasome dysfunction, mitochondrial energy metabolism deficiencies, and synaptic function [5-11]. To date no such proteomic studies have been reported in the related and rare basal ganglia disorder LND, a developmental rather than a neurodegenerative neurological disorder caused by deficiency of the enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) that regulates a major step in the purine salvage pathway [12]. Many studies have demonstrated that the both human LND patients and a mouse knockout model of HPRT deficiency have significantly reduced levels and uptake of dopamine in the striatum [4, 13-16] that is likely to be the principal cause of the CNS disorder. The precise molecular and cellular mechanisms that underlie this neurotransmitter defect are unknown. PMID- 17922513 TI - HUPO BPP pilot study: a proteomics analysis of the mouse brain of different developmental stages. AB - This study is a part of the HUPO Brain Proteome Project (BPP) pilot study, which aims at obtaining a reliable database of mouse brain proteome, at the comparison of techniques, laboratories, and approaches as well as at preparing subsequent proteome studies of neurologic diseases. The C57/Bl6 mouse brains of three developmental stages at embryonic day 16 (E16), postnatal day 7 (P7), and 8 wk (P56) (n = 5 in each group) were provided by the HUPO BPP executive committee. The whole brain proteins of each animal were individually prepared using 2-DE coupled with PDQuest software analysis. The protein spots representing developmentally related or stably expressed proteins were then prepared with in gel digestion followed with MALDI-TOF/TOF MS/MS and analyzed using the MASCOT search engines to search the Swiss-Prot or NCBInr database. The 2-DE gel maps of the mouse brains of all of the developmental stages were obtained and submitted to the Data Collection Centre (DCC). The proteins alpha-enolase, stathmin, actin, C14orf166 homolog, 28,000 kDa heat- and acid-stable phosphoprotein, 3 mercaptopyruvate sulfurtransferase and 40 S ribosomal protein S3a were successfully identified. A further Western blotting analysis demonstrated that enolase is a protein up-regulated in the mouse brain from embryonic stage to adult stage. These data are helpful for understanding the proteome changes in the development of the mouse brain. PMID- 17922514 TI - Membrane subproteomic analysis of Mycobacterium bovis bacillus Calmette-Guerin. AB - Mycobacterium bovis bacillus Calmette-Guerin (BCG) vaccine has been known for a long time to prevent tuberculosis (TB) worldwide since 1921. Nonetheless, we know little about BCG membrane proteome. In the present study, we utilized alkaline incubation and Triton X-114-based methods to enrich BCG membrane proteins and subsequently digested them using proteolytic enzyme. The recovered peptides were further separated by 2-D LC and identified by ESI-MS/MS. As a result, total 474 proteins were identified, including 78 integral membrane proteins (IMPs). Notably, 18 BCG IMPs were described for the first time in mycobacterium. Further analysis of the 78 IMPs indicated that the theoretical molecular mass distribution of them ranged from 8.06 to 167.86 kDa and pI scores ranged from 4.40 to 11.60. Functional classification revealed that a large proportion of the identified IMPs (67.9%, 53 out of 78) were involved in cell wall and cell processes functional group. In conclusion, here we reported a comprehensive profile of the BCG membrane subproteome. The present investigation may allow the identification of some valuable vaccine and drug target candidates and thus provide basement for future designing of preventive, diagnostic, and therapeutic strategies against TB. PMID- 17922515 TI - Pro-oxidative DEP chemicals induce heat shock proteins and an unfolding protein response in a bronchial epithelial cell line as determined by DIGE analysis. AB - Ambient particulate matter (PM) induces adverse health effects through the ability of pro-oxidative chemicals to induce the production of oxygen radicals and oxidant injury. Utilizing a proteomics strategy involving 2-D DIGE, immunoblotting, and real-time PCR, we demonstrate that organic diesel exhaust particle (DEP) chemicals induce an unfolding protein response (UPR) and proinflammatory effects in the human bronchial epithelial cell line, BEAS-2B. DIGE and MS showed the induction of at least 14 proteins, among which heat shock protein 70 (HSP70), HSP40, TPR2, and T-complex protein 1 (zeta-subunit) are known to play a role in the UPR. Demonstrating increased HSP70 mRNA expression and nuclear translocation of HSF1, the key transcription factor responsible for HSP expression, further strengthened this notion. Immunoblotting demonstrated increased expression of ATF4, an ER stress-associated transcriptional enhancer responsible for differential protein translation under conditions of ER stress. Finally, the DEP extract induced the expression of IL-6 and IL-8 in the culture supernatant. The role of oxidative stress was demonstrated further by response subtraction in the presence of the thiol antioxidant, N-acetyl cysteine. Our data suggest that pro-oxidative DEP chemicals induce protein unfolding/misfolding that lead to UPR and proinflammatory effects in a cell type that is targeted by PM in the lung. PMID- 17922516 TI - Comparative quantitative proteomics to investigate the remodeling of bioenergetic pathways under iron deficiency in Chlamydomonas reinhardtii. AB - The basic question addressed in this study is how energy metabolism is adjusted to cope with iron deficiency in Chlamydomonas reinhardtii. To investigate the impact of iron deficiency on bioenergetic pathways, comparative proteomics was combined with spectroscopic as well as voltametric oxygen measurements to assess protein dynamics linked to functional properties of respiratory and photosynthetic machineries. Although photosynthetic electron transfer is largely compromised under iron deficiency, our quantitative and spectroscopic data revealed that the functional antenna size of photosystem II (PSII) significantly increased. Concomitantly, stress-related chloroplast polypeptides, like 2-cys peroxiredoxin and a stress-inducible light-harvesting protein, LhcSR3, as well as a novel light-harvesting protein and several proteins of unknown function were induced under iron-deprivation. Respiratory oxygen consumption did not decrease and accordingly, polypeptides of respiratory complexes, harboring numerous iron sulfur clusters, were only slightly diminished or even increased under low iron. Consequently, iron-deprivation induces a transition from photoheterotrophic to primarily heterotrophic metabolism, indicating that a hierarchy for iron allocations within organelles of a single cell exists that is closely linked with the metabolic state of the cell. PMID- 17922517 TI - Membrane proteins from the cyanobacterium Synechocystis sp. PCC 6803 interacting with thioredoxin. AB - Cysteine dithiol/disulphide exchange forms the molecular basis for regulation of a wide variety of enzymatic activities and for transduction of cellular signals. Thus, the search for proteins with reactive, accessible cysteines is expected to contribute to the unravelling of new molecular mechanisms for enzyme regulation and signal transduction. Several methods have been designed for this purpose taking advantage of the interactions between thioredoxins and their protein substrates. Thioredoxins comprise a family of redox-active enzymes, which catalyse reduction of protein disulphides and sulphenic acids. Due to the inherent practical difficulties associated with studies of membrane proteins these have been largely overlooked in the many proteomic studies of thioredoxin interacting proteins. In the present work, we have developed a procedure to isolate membrane proteins interacting with thioredoxin by binding in situ to a monocysteinic His-tagged thioredoxin added directly to the intact membranes. Following fractionation and solubilisation of the membranes, thioredoxin target proteins were isolated by Ni-affinity chromatography and 2-DE SDS-PAGE under nonreducing/reducing conditions. Applying this method to total membranes, including thylakoid and plasma membranes, from the cyanobacterium Synechocystis sp. PCC 6803 we have identified 50 thioredoxin-interacting proteins. Among the 38 newly identified thioredoxin targets are the ATP-binding subunits of several transporters and members of the AAA-family of ATPases. PMID- 17922519 TI - Electrokinetic injection of DNA from gel micropads: basis for coupling polony technology with CE separation. AB - We propose a novel method for electrokinetic injection of DNA samples into capillaries from nanoliter gel micropads, deposited on glass slides, which are coated with electroconducting film. Theoretical and experimental proof is presented for the proposed method. The method allows efficient and highly precise injection without physical contact between the gel pad and the capillary. Read length of more than 700 bp at Q20 has been reproducibly demonstrated in fused silica capillaries using the proposed injection technique. Based on the obtained results we discuss a novel DNA sequencing system which combines DNA amplification and cycle sequencing in arrays of subnanoliter gel micropads and high-throughput electrophoretic separation in monolith multicapillary arrays. PMID- 17922520 TI - Chiral resolution of melatoninergic ligands by EKC using highly sulfated CDs. AB - EKC methods for the enantiomeric resolutions of melatoninergic ligands were developed using anionic CDs (highly S-alpha-CD, highly S-beta-CD, and highly S gamma-CD) as chiral selectors at acidic pH 2.5. The optimization of the various operational parameters (nature and concentration of the CD, phosphate buffer concentration, addition of organic modifiers in the BGE, and temperature) allows baseline enantioresolutions (superior to 2) in short analysis times (inferior to 7 min) for all studied analytes. Some analytical characteristics of the optimal method were then studied for each analyte: repeatability, linearity, and LOD and LOQ. Lastly, determination of the apparent binding constants for the 18 complexes formed between the six analytes and the three CDs led us to rationalize the complexation mechanisms. PMID- 17922518 TI - Tandem mass spectrometry for the detection of plant pathogenic fungi and the effects of database composition on protein inferences. AB - LC-MS/MS has demonstrated potential for detecting plant pathogens. Unlike PCR or ELISA, LC-MS/MS does not require pathogen-specific reagents for the detection of pathogen-specific proteins and peptides. However, the MS/MS approach we and others have explored does require a protein sequence reference database and database-search software to interpret tandem mass spectra. To evaluate the limitations of database composition on pathogen identification, we analyzed proteins from cultured Ustilago maydis, Phytophthora sojae, Fusarium graminearum, and Rhizoctonia solani by LC-MS/MS. When the search database did not contain sequences for a target pathogen, or contained sequences to related pathogens, target pathogen spectra were reliably matched to protein sequences from nontarget organisms, giving an illusion that proteins from nontarget organisms were identified. Our analysis demonstrates that when database-search software is used as part of the identification process, a paradox exists whereby additional sequences needed to detect a wide variety of possible organisms may lead to more cross-species protein matches and misidentification of pathogens. PMID- 17922521 TI - Molecular deformation and free-solution electrophoresis of DNA-uncharged polymer conjugates at high field strengths: theoretical predictions Part 2: Stretching. AB - DNA sequencing by electrophoresis can be dramatically sped up by overcoming the need for the sieving medium. Normally it is possible to separate DNA based on size in free solution; however, not end-labeled free-solution electrophoresis (ELFSE) uses a neutral drag-tag molecule to make it possible. In experiments to date, the drag-tag and DNA together form a random coil conformation; while with future generation drag-tags and high fields, deformation of this conformation may occur. In the first paper in this series we investigated the conditions under which the DNA and label become hydrodynamically distinct (or segregated), based on a theoretical approach developed for the electrophoresis of polyampholytes. In this paper we study further deformation wherein either the DNA and/or a polymeric label stretch. We show that deformation may dramatically improve the capabilities of ELFSE, especially when both the DNA and a polymeric drag-tag fully stretch; however, reaching these regimes will require extremely high field intensities, something that only microchip technologies may be able to achieve. PMID- 17922522 TI - Improved solubilization of surface proteins from Listeria monocytogenes for 2-DE. AB - Solubilization of bacterial surface (cell wall and membrane-associated) proteins for 2-DE is challenging, particularly in the case of Gram-positive bacteria. This is primarily due to strong protein association with the cell wall peptidoglycan and protein hydrophobicity. We solubilized surface proteins for 2-DE from the Gram-positive pathogen Listeria monocytogenes using mutanolysin, which digests cell wall peptidoglycan, and one of three different mixtures of zwitterionic detergent and chaotropes: (i) CHAPS/urea, (ii) amidosulfobetaine-14 (ASB 14)/urea/thiourea (iii) N-decyl-N,N'-dimethyl-3-ammonio-1 propanesulfonate/urea/thiourea. Cell lysis with mutanolysin followed by solubilization with ASB-14/urea/thiourea gave the highest overall protein yield with the best 2-DE resolution. Protein spot identification by MALDI-TOF/TOF-MS analysis revealed 29 characterized surface proteins of L. monocytogenes, 17 of which have not previously been reported on the surface proteome map. This is the first report describing the successful solubilization and 2-DE of L. monocytogenes proteins bound to the cell surface via an LPXTG motif or by a hydrophobic tail. The increase in surface proteome coverage obtained by mutanolysin and ASB-14/urea/thiourea solubilization suggests the utility of this method for future analytical and comparative studies of surface proteins from Listeria, and possibly other Gram-positive bacteria, using 2-DE proteomic analysis. An updated 2-DE reference map of L. monocytogenes surface proteins is presented. PMID- 17922523 TI - Whole-brain radiotherapy versus stereotactic radiosurgery for patients in recursive partitioning analysis classes 1 and 2 with 1 to 3 brain metastases. AB - BACKGROUND: The authors investigated whether stereotactic radiosurgery (SRS) alone improved outcomes for patients in recursive partitioning analysis (RPA) Classes 1 and 2 who had 1 to 3 brain metastases compared with whole-brain radiotherapy (WBRT). METHODS: Data regarding 186 patients in RPA Classes 1 and 2 who had 1 to 3 brain metastases and who received either 30 to 40 grays (Gy) of WBRT (n = 91 patients) or 18 to 25 Gy SRS (n = 95 patients) were analyzed retrospectively. Eight other potential prognostic factors were evaluated regarding overall survival (OS), entire brain control (BC), local control (LC) of treated metastases, and brain control distant from treated metastases (distant control [DC]): Those 8 factors were age, sex, performance status, tumor type, number of brain metastases, extracranial metastases, RPA class, and interval from tumor diagnosis to radiotherapy. RESULTS: On multivariate analysis of OS, age ( risk ratio [RR], 1.51; P = .024), Karnofsky performance status (KPS) (RR, 1.98; P = .002), and extracranial metastases (RR, 2.26; P < .001) were significant, whereas the radiation regimen was not significant (P = .89). On multivariate analysis of BC, only the radiation regimen (RR, 1.33; P = .003) was found to be significant. On multivariate analysis of LC, radiation regimen (RR, 1.63; P < .001) and sex (RR, 1.62; P = .022) were significant. On multivariate analysis of DC, KPS (RR, 1.85; P = .049) and extracranial metastases (RR, 1.69; P = .047) were significant. The radiation regimen was not found to be significant even on univariate analysis (P = .80). In RPA class subgroup analyses, BC and LC were better after SRS than WBRT for patients in RPA Classes 1 and 2, whereas OS and DC did not differ significantly. CONCLUSIONS: For patients in RPA Classes 1 and 2 who had 1 to 3 brain metastases, SRS alone was associated with improved BC and LC compared with 30 to 40 Gy WBRT, whereas OS and DC were not significantly different. Similar results were observed in separate subgroup analyses of patients in RPA Class 1 and RPA Class 2. PMID- 17922525 TI - Estimation of treatment effect adjusting for treatment changes using the intensity score method: application to a large primary prevention study for coronary events (MEGA study). AB - The MEGA study was a prospective, randomized, open-labeled, blinded-endpoints study conducted in Japan to evaluate the primary preventive effect of pravastatin against coronary heart disease (CHD), in which 8214 subjects were randomized to diet or diet plus pravastatin. The intention-to-treat (ITT) analysis showed that pravastatin reduced the incidence of CHD (hazard ratio=0.67; 95 per cent confidence interval (CI): 0.49-0.91) and of stroke events, which was the secondary endpoint in the MEGA study (hazard ratio=0.83; 95 per cent CI: 0.57 1.21). Owing to considerable treatment changes, it is also of interest to estimate the causal effect of treatment that would have been observed had all patients complied with the treatment to which they were assigned. In this paper, we present an intensity score method developed for clinical trials with time-to event outcomes that correct for treatment changes during follow-up. The proposed method can be easily extended to the estimation of time-dependent treatment effects, where the technique of g-estimation has been difficult to apply in practice. We compared the performances of the proposed method with other methods (as-treated, ITT, and g-estimation analysis) through simulation studies, which showed that the intensity score estimator was unbiased and more efficient. Applying the proposed method to the MEGA study data, several prognostic factors were associated with the process of treatment changes, and after adjusting for these treatment changes, larger treatment effects for pravastatin were observed for both CHD and stroke events. The proposed method provides a valuable and flexible approach for estimating treatment effect adjusting for non-random non compliance. PMID- 17922527 TI - Note on drill (Mandrillus leucophaeus) ecology and conservation status in Korup National Park, Southwest Cameroon. AB - We report preliminary findings on drill population ecology, feeding ecology, primate associations and conservation status in Korup National Park, Cameroon, based on analysis of data collected during 1,346 km (620 field hours) of trail patrols from February to June 2006. We encountered drills on 25 occasions and collected 304 fecal samples. Group size was estimated on four occasions (> or =25, > or =40, > or =67, > or =77), and multiple males were heard emitting the characteristic two-phase grunt of mature male drills in two of these groups. We saw a solitary male drill once. Examination of fecal samples indicated a seasonally variable diet consisting mainly of seeds, fruit pith, leaves and insects (especially ants and termites). Irvingia gabonensis and Musanga cecropioides fruits showed the highest percentage of occurrence in May (96%) and March (75%) samples, respectively, and could be a major food source for drills at that time. Drills were in association with at least one additional primate species during 57.9% of observations. These associations involved all of the diurnal primates found in Korup (Cercopithecus mona, Cercopithecus nictitans, Cercopithecus erythrotis, Cercopithecus pogonias, Piliocolobus preussi and Cercocebus torquatus) except chimpanzees (Pan troglodytes). We discuss our findings in terms of existing knowledge about drill ecology and highlight the urgent need for the protection of the significant drill population in Korup National Park. PMID- 17922526 TI - Predictors of injurious assault committed during or after drinking alcohol: a case-control study of young offenders. AB - Studies of causal links between alcohol and aggression are often handicapped by threats to internal and external validity. Case-control methods employ an event level analysis that can reduce some of these validity threats by the use of within-subject controls. This study used a case-control approach, asking 39 male inmates in a Young Offenders' Institution to compare drinking behaviour before incidents where they reported commission of an injurious assault and a matched incident where they did not. After controlling contextual differences, participants reported personally drinking more heavily and heavier drinking within their group, but not being more impaired when an assault was committed. The assault incidents were more likely to involve spontaneous, rather than planned, drinking and a higher proportion of males in the group. They were less likely to involve drinking in a pub. Our confirmation of previous findings using a case-control methodology strengthens those findings. Limitations of this methodology are also discussed. PMID- 17922528 TI - TC-tuned biocompatible suspension of La0.73Sr0.27MnO3 for magnetic hyperthermia. AB - La(1-x)Sr(x)MnO(3), a ferromagnet with high magnetization and Curie temperature T(C) below 70 degrees C, enables its use for magnetic hyperthermia treatment of cancer with a possibility of in vivo temperature control. We found that La(0.73)Sr(0.27)MnO(3) particles of size range 20-100 nm showed saturation magnetization around 38 emu/g at 20 kOe and a T(C) value of 45 degrees C. Aqueous suspension of these nanoparticles was prepared using a polymer, acrypol 934, and the biocompatibility of the suspension was examined using HeLa cells. A good heating ability of the magnetic suspension was obtained in the presence of AC magnetic field, and it was found to increase with the amplitude of field. The suspension having concentration of 0.66 mg/mL (e.g., 0.66 mg of nanoparticles with acropyl per milliliter of culture media) was observed to be biocompatible even after 96 h of treatment, as estimated by sulforhodamine B and trypan blue dye exclusion assays. Further, the treatment with the aforementioned concentration did not alter the microtubule cytoskeleton or the nucleus of the cells. However, the bare particles (concentration of 0.66 mg of nanoparticles per milliliter of culture media, but without acropyl) decreased the viability of cell significantly. Our in vitro studies suggest that the suspension (concentration of 0.66 mg/mL) may further be analyzed for in vivo studies. PMID- 17922529 TI - Impact of lowering the criterion for impaired fasting glucose on identification of individuals with insulin resistance. The GISIR database. AB - OBJECTIVE: We assessed the accuracy of the American Diabetes Association (ADA)2003 definition of impaired fasting glucose (IFG) in identifying subjects with low insulin sensitivity, and determined cardiovascular risk factors in ADA2003 IFG subjects. RESEARCH DESIGN AND METHODS: This study included 930 non diabetic Italian Caucasians from the GISIR database in which subjects underwent a hyperinsulinaemic-euglycaemic clamp performed with a standard technique. Low insulin sensitivity was defined as being in the lower quartile of glucose metabolized during the last hour of the clamp (M). Subjects were stratified in the following groups: normal fasting glucose (NFG) (<100 mg/dL), IFG100 (100-109 mg/dL), ADA1997 IFG110 (110-125 mg/dL), and ADA2003 IFG (100-125 mg/dL). RESULTS: The sensitivity of identifying subjects with low insulin sensitivity increased adopting the ADA2003 criterion. After Bonferroni correction for multiple comparisons, both IFG100 and ADA1997 IFG110 showed significantly higher body mass index (BMI), waist, systolic blood pressure (SBP) and diastolic blood pressure (DBP), triglyceride, fasting plasma insulin (FPI) and fasting plasma glucose (FPG), and lower insulin sensitivity as compared with NFG. As compared with IFG100, ADA1997 IFG110 showed significantly higher BMI, waist, SBP, FPI, FPG, and lower insulin sensitivity. ADA2003 IFG group showed significantly higher BMI, waist, SBP and DBP, triglyceride, cholesterol, FPI, and FPG, but lower HDL levels and insulin sensitivity compared with NFG subjects. CONCLUSIONS: Although neither the ADA2003 nor the ADA1997 definition of IFG appears to be particularly efficacious for the identification of subjects' low insulin sensitivity, lowering the criterion to the ADA2003 glucose threshold increased the sensitivity without affecting the specificity. ADA2003 IFG showed a worse cardiovascular risk profile compared with NFG. PMID- 17922530 TI - A DRD4/BDNF gene-gene interaction associated with maximum BMI in women with bulimia nervosa. AB - OBJECTIVE: The goals of the current study were threefold: 1) to examine whether the hypofunctional 7R allele of the DRD4 gene contributes to maximal lifetime body mass in women with BN; 2) to determine whether the BDNF gene contributes to maximal BMI on its own, and 3) to explore possible BDNF/DRD4 gene-gene interactions in mediating maximum lifetime BMIs in BN. METHOD: We tested two General Linear Models predicting maximum lifetime BMI with the exon 3 VNTR polymorphism of the dopamine-4 receptor gene (DRD4) and either the Val66Met or the -270C/T polymorphism of BDNF respectively in 163 female probands with BN, purging subtype. RESULTS: In these bulimic subjects, the hypofunctional 7R allele of DRD4 predicted maximal BMI (p < .01). There was also a significant interaction between the DRD4 gene and the BDNF gene in predicting maximal BMI. The Val66Met rather than the 270C/T polymorphism of BDNF interacting with DRD4 predicted maximum BMI in this BN sample (p < .01). Probands carrying both the hypofunctional 7R allele of DRD4 and the Met66 allele of BDNF had significantly higher maximal BMI than did probands in the other gene-gene interaction groups. CONCLUSION: These results provide further evidence that the hypofunctional 7R allele of DRD4 contributes to weight gain in women with BN and that the BDNF gene interacts with DRD4 to influence weight regulation in these subjects. PMID- 17922531 TI - Eating problems, body image disturbances, and academic achievement: preliminary evaluation of the eating and body image disturbances academic interference scale. AB - OBJECTIVE: To examine the relationships between a new scale, the Eating and Body Image Disturbances Academic Interference Scale (EBIDAIS), and measures of eating disturbance, body image, and academic achievement. METHOD: One thousand five hundred eighty-four college undergraduates completed the measures in an online survey and were awarded class credit for their participation. Measures included the Eating Disorder Inventory Bulimia, Drive for Thinness, Body Dissatisfaction, and Perfectionism subscales. Grade point average (GPA) was also reported. RESULTS: Academic interference and GPA were significantly correlated, indicating that higher interference scores were related to lower GPA. EBIDAIS was also significantly correlated with drive for thinness, bulimia, and body dissatisfaction, but was not significantly associated with perfectionism. The correlation between interference and GPA was substantially higher for a subsample of individuals who scored in the elevated range on eating and body dissatisfaction. CONCLUSION: Academic interference may be a relatively unexamined, but potentially important, outcome for individuals who experience eating problems and body image disturbance. PMID- 17922532 TI - Expressed emotion in eating disorders assessed via self-report: an examination of factors associated with expressed emotion in carers of people with anorexia nervosa in comparison to control families. AB - OBJECTIVE: High expressed emotion (EE) has been linked to outcome in anorexia nervosa (AN). Traditional interview measures of EE require considerable resources. The aim was to compare EE, measured with a self report tool, in parents of people with AN (carers; N = 151) with that of parents of healthy people (controls; N = 93), and to explore factors associated with EE. METHOD: A cross-sectional design was used to assess EE (Family Questionnaire) and psychological distress (Hospital Anxiety and Depression Scale). RESULTS: Over 60% (96/151) of carers showed high emotional over-involvement (EOI) compared with 3% (3/93) of controls, whereas 47% (71/151) of carers showed high criticism (CC) compared with 15% (14/93) of controls. The most significant variable related to CC was "negative/difficult behaviors," and anxiety, for EOI. CONCLUSION: The FQ was sensitive both to CC and EOI. Interventions aimed at reducing EE may need to focus on decreasing carers' anxiety and teaching skills to moderate difficult behaviors. PMID- 17922533 TI - A critical evaluation of the efficacy of self-help interventions for the treatment of bulimia nervosa and binge-eating disorder. AB - OBJECTIVE: Cognitive behavioral therapy (CBT) is efficacious for the treatment of bulimia nervosa (BN) and binge-eating disorder (BED). As a number of factors limit the availability of CBT, self-help manuals have been developed to make the treatment more widely available. METHOD: Published studies evaluating the efficacy of self-help programs in the treatment of BN and BED were reviewed. RESULTS: Controlled studies of self-help programs for BN and BED have often employed a waiting list control group, and indicate that self-help provides more benefit than remaining on a waiting list. However, fewer studies have utilized a more active control group, and these studies have not been as positive. CONCLUSION: In general, open and wait-list trials indicate that self-help is helpful in treating BN and BED, but there is little evidence for the specific efficacy of self-help in comparison to other treatments. Additional studies of self-help are needed to determine the specific utility of self-help interventions for BN and BED. PMID- 17922535 TI - Effects of dieting-related messages on psychological and weight control variables. AB - OBJECTIVE: The current study sought to empirically examine the immediate and short-term effects of dieting-related psychoeducational messages on established risk factors for eating pathology and weight control variables. METHOD: One hundred thirty nine participants were randomly assigned to a prodieting, antidieting, or control message condition. Variables were assessed at baseline, post-test, and 2-week follow-up. RESULTS: The prodieting condition produced significantly greater dieting intentions, perceived pressure to lose weight, and internalization intentions, whereas the antidieting message yielded significantly lower bulimic intentions. Healthy eating significantly increased from baseline to follow-up in the prodieting condition. No other behavioral changes were found. Perceived pressure to lose weight mediated the relationship between diet message and post-test weight control intentions, state negative-affect, and body dissatisfaction. CONCLUSION: The findings provide initial support for the short term efficacy of the prodieting message to increase healthy eating behaviors; however, immediate increases in established risk factors for eating pathology also emerged. Implications are discussed. PMID- 17922534 TI - PANDAS anorexia nervosa--endangered, extinct or nonexistent? AB - OBJECTIVE: Substantial evidence exists for the concept of pediatric autoimmune neuropsychiatric disorders associated with streptoccoccal infection (PANDAS) such as some cases of obsessive-compulsive disorder and tics/Tourette's syndrome. More recently PANDAS-AN has been described. The aim of this article is to provide a critical review of this concept. METHOD: The literature was searched using Medline, Psychinfo, and Google. RESULTS: There is some evidence for the entity of PANDAS-AN but this is considerably limited by a wide range of methodological problems, including the seemingly low prevalence, the lack of clarity in diagnostic criteria, and the lack of specificity in the diagnostic tests. Innovative treatment approaches hold promise but require further evaluation. CONCLUSION: Further research into PANDAS-AN, using more clearly defined and consistent diagnostic criteria is needed, given the potential for enhancing the understanding of the pathogenesis of AN and for more effective treatments. PMID- 17922536 TI - Body composition in girls and young women with anorexia nervosa: comparison of different equations. AB - OBJECTIVE: The weight percentage of body fat in anorexic adolescents on the basis of four distinct formulas (Siri; Lohman; Westrate and Deurenberg; Heyward and Stolarczyk) is compared. METHOD: The body composition of 238 anorexia nervosa (AN) patients, divided into four age categories (13-15, 15-17, 17-19, and 19-22 years), was measured by means of densitometry (underwater weighing). RESULTS: Depending on the formula, the results calculated by the formulas differ significantly in each age category. The Siri formula generally results in the highest mean fat percentage (12.76%-13.39%) whereas the Heyward and Stolarczyck formula shows the lowest figures (8.77%-9.31%). Applying the Lohman and the Heyward and Stolarczyck formulas to the 19-22 years category, results in negative fat percentages. CONCLUSION: A clinically useful formula is important for the estimation of body composition in specific age categories. We recommend with some restrictions, to apply the Siri formula in AN patients. PMID- 17922537 TI - The diagnostic validity of the criteria for binge eating disorder. AB - OBJECTIVE: This paper considers whether the criteria currently used to classify the diagnosis of binge eating disorder (BED) are valid and appropriate. METHOD: We review evidence that reflects on the validity of the current criteria for binge eating episodes and BED, using literature retrieved through major psychology and psychiatry search engines (e.g., PsycInfo, PubMed). RESULTS: Evidence from experimental research points to the relative importance of episode frequency, the amount of food consumed at episodes, the subjective sense of loss of control over eating, and several additional criteria associated with binge episodes in BED. Evidence on the differences in psychopathology between BED and bulimia nervosa and between BED and obesity without binge eating, as related to diagnostic criteria, is reviewed. CONCLUSION: Although evidence concerning the diagnostic criteria of BED is mixed, broadening certain diagnostic criteria for binge eating episodes and BED might more accurately reflect the research literature and increase the number of individuals eligible for inclusion in treatment programs. PMID- 17922538 TI - Suicidal behavior in adolescents: relationship to weight status, weight control behaviors, and body dissatisfaction. AB - OBJECTIVE: Suicide is associated with full syndromal eating disorders, but it is unclear whether subsyndromal eating disorders carry the same risk. This study examined associations between suicidal behaviors and extreme and less extreme weight control behaviors (EWCB and LWCB, such as fasting, vomiting, meal skipping, etc.), body dissatisfaction, and weight status in adolescents. METHOD: Data on body dissatisfaction, depressive symptoms, suicidal ideation and attempts, and body mass index (BMI) were drawn from Project EAT, a survey of 4,746 7th-12th grade students. Multivariate logistic regression examined associations between eating- and weight-related variables and suicidality. RESULTS: Suicidal ideation and suicide attempts were more commonly observed in adolescents with EWCB (boys: ideation OR = 2.12, attempts OR = 4.10; girls: ideation OR = 1.66, attempts OR = 2.29), LWCB (boys: ideation OR = 1.33, attempts OR = 1.76; girls: ideation OR = 1.77, attempts OR = 1.80), as well as body dissatisfaction (boys: ideation OR = 1.75, attempts OR = 2.23; girls: ideation OR = 1.77, attempts OR = 1.81), even after controlling for depressive symptoms. No association was observed between BMI and suicidal attempts or ideation. CONCLUSION: Thus, it appears that suicidal behavior in adolescents is associated even with low-level eating disorder symptoms. PMID- 17922539 TI - Effects of physiological fluids on physical-chemical characteristics and activity of topical vaginal microbicide products. AB - The increased incidence of HIV infection in women has identified a need to develop a female controlled method to prevent sexually transmitted infections (STIs), including HIV. Formulations have been developed in our laboratory for two potential microbicide drug substances, 3-O-octyl-sn-glycerol (3-OG) and UC-781. A major concern for microbicide product development is dilution by vaginal fluids following application thereby reducing antimicrobial activity. We investigated the effect of product dilution on microbicidal activity and the product's chemical and physical properties by using vaginal fluid (VFS) and cervical mucus simulants (CMS). 3-OG and UC-781 were individually formulated into three semi solid drug containing products: Hydroxyethylcellulose, Methylcellulose/Carbopol, and Liposome. Viscosity, osmolality, pH and in vitro activity against HIV-1 were evaluated. Results showed that pH was not affected when products were diluted with VFS; however, increases in pH were observed following CMS dilution. Viscosity was significantly decreased for all the dilutions tested excepted for some of the liposome products. Hydrogel products maintained greater activity against HIV-1 than Liposome products. The effect of dilution on anti-HIV activity varied based upon excipient choice and chemical characteristics of the active agent. These in vitro assessments can identify the potential for changes in product's physical-chemical characteristics in vivo which may result in diminished product performance. PMID- 17922540 TI - Health pioneers. PMID- 17922541 TI - You're the flight surgeon. Carbon monoxide poisoning. PMID- 17922542 TI - Dr. Ronald B. Gross receives Pierre Fauchard Award. PMID- 17922543 TI - Health care cost and access challenges persist: initial findings from HSC's 2007 site visits. AB - Little has changed in local health care markets since 2005 to break the cycle of rising costs, falling insurance coverage and widening access inequities, according to initial findings from the Center for Studying Health System Change's (HSC) 2007 site visits to 12 nationally representative metropolitan communities. As intense competition among hospitals and physicians for profitable specialty services continues, employers and health plans are looking to consumers to take more responsibility for medical costs, lifestyle choices and treatment decisions. While consumer-directed health plans have not gained widespread adoption, other developments-including a heightened emphasis on prevention and wellness, along with nascent provider cost and quality information-are advancing health care consumerism. However, concerns exist about whether these efforts will slow cost growth enough to keep care affordable or whether the growing problem of affordability will derail efforts to decrease the rising number of uninsured Americans and stymie meaningful health care reform. PMID- 17922544 TI - Insulin receptor substrate 1 knockdown in human MCF7 ER+ breast cancer cells by nuclease-resistant IRS1 siRNA conjugated to a disulfide-bridged D-peptide analogue of insulin-like growth factor 1. AB - IRS-1 overexpression has been associated with breast cancer development, hormone independence and antiestrogen resistance. IRS-1 is a major downstream signaling protein for insulin and IGF1 receptors, conveying signals to PI-3K/Akt and ERK1/2 pathways. In estrogen-sensitive breast cancer cell lines, the widely used antiestrogen tamoxifen treatment reduces IRS-1 expression and function, thereby inhibiting IRS-1/PI-3K signaling. IRS-1 may serve as an alternative target to overexpressed IGF1R in breast cancer. While siRNA technology has become commonplace in many laboratories for in vitro gene knockdown studies, and in vivo stability issues are largely solved, its use in vivo is limited by an inability to efficiently and specifically deliver it to the intended site of action. We previously reported reduced survival of human MCF7 estrogen receptor positive breast cancer cells treated with a normal IRS1 siRNA delivered by a cationic lipid, plus an additive effect in combination with tamoxifen. We now report enhanced cellular uptake, relative to the unconjugated serum-stabilized IRS1 siRNA, of a serum-stabilized IRS1 siRNA conjugated with our previously characterized peptide mimetic of IGF1, D-(Cys-Ser-Lys-Cys), without the use of cationic lipids or electroporation, in MCF7 cells that overexpress IGF1R. Excess native IGF1 blocked uptake. An IRS1 siRNA cholesterol conjugate, targeted universally to cell membranes, was taken up by MCF7 cells as much as the peptide mimetic conjugate. IRS1 mRNA knockdown and IRS-1 protein knockdown were comparable for the IGF1 peptide and cholesterol conjugates. The unconjugated serum-stabilized IRS1 siRNA control showed negligible effects. Viability assays showed additive effects of siRNA treatment in combination with tamoxifen. In summary, we have taken the first step in converting an siRNA into a pharmacologically active agent for breast cancer. PMID- 17922545 TI - Oligonucleotide-coated metallic nanoparticles as a flexible platform for molecular imaging agents. AB - Targeted metallic nanoparticles have shown promise as contrast agents for molecular imaging. To obtain molecular specificity, the nanoparticle surface must be appropriately functionalized with probe molecules that will bind to biomarkers of interest. The aim of this study was to develop and characterize a flexible approach to generate molecular imaging agents based on gold nanoparticles conjugated to a diverse range of probe molecules. We present two complementary oligonucleotide-based approaches to develop gold nanoparticle contrast agents which can be functionalized with a variety of biomolecules ranging from small molecules, to peptides, to antibodies. The size, biocompatibility, and protein concentration per nanoparticle are characterized for the two oligonucleotide based approaches; the results are compared to contrast agents prepared using adsorption of proteins on gold nanoparticles by electrostatic interaction. Contrast agents prepared from oligonucleotide-functionalized nanoparticles are significantly smaller in size and more stable than contrast agents prepared by adsorption of proteins on gold nanoparticles. We demonstrate the flexibility of the oligonucleotide-based approach by preparing contrast agents conjugated to folate, EGF peptide, and anti-EGFR antibodies. Reflectance images of cancer cell lines labeled with functionalized contrast agents show significantly increased image contrast which is specific for the target biomarker. To demonstrate the modularity of this new bioconjugation approach, we use it to conjugate both fluorophore and anti-EGFR antibodies to metal nanoparticles, yielding a contrast agent which can be probed with multiple imaging modalities. This novel bioconjugation approach can be used to prepare contrast agents targeted with biomolecules that span a diverse range of sizes; at the same time, the bioconjugation method can be adapted to develop multimodal contrast agents for molecular imaging without changing the coating design or material. PMID- 17922547 TI - Preparation and evaluation of glycosylated arginine-glycine-aspartate (RGD) derivatives for integrin targeting. AB - Arginine-glycine-aspartate (RGD) derivatives were prepared by a combination of solid-phase and solution-phase synthesis for selective targeting of alpha vbeta 3 integrin expressed in tumors. In order to evaluate the value of a triazole moiety as a proposed amide isostere, the side chain glycosylated cyclic RGD ( cRGD) peptides were synthesized with either a natural amide linkage or a triazole. Affinity of the cRGD constructs for the alpha vbeta 3 integrin was determined in a solid-phase competitive binding assay, showing strong similarity in binding affinity for each of the compounds under evaluation. Furthermore, the in vivo tumor targeting potential of glycosylated cRGD peptides, linked via amide or triazole, was investigated by determining the biodistribution of (125)I-labeled derivatives in mice with tumors expressing alpha vbeta 3. All of the cyclic RGD derivatives showed preferential uptake in the subcutaneous tumors, with the highest tumor-to-blood ratio measured for the triazole-linked glycosylated derivative. The results of the present study are a clear indication of the value of the triazole moiety as a suitable amide isostere in the development of glycosylated peptides as pharmaceuticals. PMID- 17922546 TI - Synthesis, analysis, in vitro characterization, and in vivo disposition of a lamivudine-dextran conjugate for selective antiviral delivery to the liver. AB - A liver-selective prodrug (3TCSD) of the antiviral drug lamivudine (3TC) was developed and characterized. 3TC was coupled to dextran ( approximately 25 kDa) using a succinate linker, and the in vitro and in vivo behavior of the conjugate was studied using newly developed size-exclusion and reversed-phase analytical methods. Synthesized 3TCSD had a purity of >99% with a degree of substitution of 6.5 mg of 3TC per 100 mg of the conjugate. Furthermore, the developed assays were precise and accurate in the concentration ranges of 0.125-20, 0.36-18, and 1-50 microg/mL for 3TC, 3TC succinate (3TCS), and 3TCSD, respectively. In vitro, the conjugate slowly released 3TC in the presence of rat liver lysosomes, whereas it was stable in the corresponding buffer. In vivo in rats, conjugation of 3TC to dextran resulted in 40- and 7-fold decreases in the clearance and volume of distribution of the drug, respectively. However, the accumulation of the conjugated 3TC in the liver was 50-fold higher than that of the parent drug. The high accumulation of the conjugate in the liver was associated with a gradual and sustained release of 3TC in the liver. These studies indicate the feasibility of the synthesis of 3TCS-dextran and its potential use for the selective delivery of 3TC to the liver. PMID- 17922548 TI - Synthesis and in vitro evaluation of novel star-shaped block copolymers (blocked star vectors) for efficient gene delivery. AB - Novel 4-branched diblock copolymers consisting of cationic chains as an inner domain and nonionic chains as an outer domain were prepared by iniferter-based living radial polymerization and evaluated as a polymeric transfectant. The cationic polymerization of 3-(N,N-dimethylamino)propyl acrylamide (DMAPAAm) using 1,2,4,5-tetrakis( N,N-diethyldithiocarbamylmethyl)benzene as a 4-functional iniferter followed by the nonionic block polymerization of N,N-dimethylacrylamide (DMAAm) afforded 4-branched diblock copolymers with controlled compositions. By changing the solution or irradiation conditions, 4-branched PDMAPAAms with molecular weights of 10,000, 20,000, and 50,000 were synthesized. In addition, by graft polymerization, PDMAPAAm-PDMAAm blocked copolymers with copolymer composition (unit ratio of DMAAm/DMAPAAm) ranging from 0.18 to 1.0 for each cationic polymer were synthesized. All polymers were shown to interact with and condense plasmid DNA to yield polymer/DNA complexes (polyplexes). A transfection study on COS-1 cells showed that the polyplexes from block copolymers with cationic chain length of approximately 50,000 and a nonionic chain length of 30,000, which were approximately 200 nm in diameter and very stable in aqueous media, had the most efficient luciferase activity with minimal cellular cytotoxicity under a charge ratio of 20 (vector/pDNA). The PDMAPAAm-PDMAAm blocked, star-shaped polymers are an attractive novel class of nonviral gene delivery systems. PMID- 17922549 TI - Cinnamic ester derivatives from Oxalis pes-caprae (Bermuda buttercup). AB - Seven new cinnamic ester derivatives ( 1- 7) were isolated from a methanol extract of the fresh leaves and twigs of Oxalis pes-caprae (Bermuda buttercup). The structures of these new compounds were determined by spectroscopic data interpretation. The effects of compounds 1- 7 on the germination and growth of Lactuca sativa (lettuce) were studied. PMID- 17922550 TI - Antioxidant flavonoid glycosides from aerial parts of the fern Abacopteris penangiana. AB - Five new flavan-4-ol glycosides, abacopterins E-I ( 5- 9), and seven known flavonoid glycosides ( 3 and 10- 15) were isolated from the aerial parts of the fern Abacopteris penangiana. Their structures were elucidated on the basis of extensive spectroscopic analysis, including HSQC, HMBC, (1)H- (1)H COSY, and ROESY, and chemical evidence. The isolated glycosides were evaluated for their antioxidant activity using the TEAC assay, and compounds 3, 5- 8, 10, 11, 14, and 15 showed TEAC values of 1.03-1.91 mM. PMID- 17922551 TI - Amphidinolides B6 and B7, cytotoxic macrolides from a symbiotic dinoflagellate Amphidinium species. AB - Two 26-membered macrolides, amphidinolides B6 ( 2) and B7 ( 1), have been isolated from a marine symbiotic dinoflagellate Amphidinium sp., and the structures were elucidated on the basis of detailed analyses of 2D NMR data. The relative and absolute configurations for 1 and 2 were assigned by comparison of NMR data and CD data with those of known amphidinolides. PMID- 17922552 TI - Antiprotozoal polyacetylenes from the Tanzanian medicinal plant Cussonia zimmermannii. AB - From the petroleum ether extract of the root bark of Cussonia zimmermannii four polyacetylenes, 1- 4, were isolated, three of which ( 1- 3) were active against Trypanosoma brucei rhodesiense, Trypanosoma cruzi, Plasmodium falciparum, and Leishmania donovani. PMID- 17922553 TI - Phenol-induced in vivo oxidative stress in skin: evidence for enhanced free radical generation, thiol oxidation, and antioxidant depletion. AB - A variety of phenolic compounds are utilized in industry (e.g., for the production of phenol (PhOH)-formaldehyde resins, paints and lacquers, cosmetics, and pharmaceuticals). They can be toxic to skin, causing rash, dermal inflammation, contact dermatitis, depigmentation, and cancer promotion. The biochemical mechanisms for the dermal toxicity of phenolic compounds are not well understood. We hypothesized that topical PhOH exposure results in the generation of radicals, possibly via redox-cycling of phenoxyl radicals, which may be an important contributor to dermal toxicity via the stimulation of the induction and release of inflammatory mediators. To test this hypothesis, we (1) monitored in vivo the formation of PBN-spin-trapped radical adducts by ESR spectroscopy, (2) measured GSH, protein thiols, vitamin E, and total antioxidant reserves in the skin of B6C3F1 mice topically treated with PhOH, and (3) compared the responses with those produced by PhOH in mice with diminished levels of GSH. We found that dermal exposure to PhOH (3.5 mmol/kg, 100 microL on the shaved back, for 30 min) caused oxidation of GSH and protein thiols and decreased vitamin E and total antioxidant reserves in skin. The magnitude of the PhOH-induced generation of PBN spin-trapped radical adducts in the skin of mice with diminished levels of GSH (pretreated with BCNU, an inhibitor of glutathione reductase, or BSO, an inhibitor of gamma-glutamylcysteine synthetase) was markedly higher compared to radical generation in mice treated with PhOH alone. Topical exposure to PhOH resulted in skin inflammation. Remarkably, this inflammatory response was accelerated in mice with a reduced level of GSH. Epidermal mouse cells exposed to phenolic compounds showed the induction of early inflammatory response mediators, such as prostaglandin E 2 and IL-1beta. Since dermal exposure to PhOH produced ESR-detectable PBN spin-trapped signals of lipid-derived radicals, we conclude that this PhOH-induced radical formation is involved in oxidative stress and dermal toxicity in vivo. PMID- 17922554 TI - A general method for designing combinatorial peptide libraries decodable by amino acid analysis. AB - Herein we describe an algorithm for designing combinatorial peptide libraries for split-and-mix synthesis on solid support that are decodable by amino acid analysis (AAA) of the beads. AAA is a standard service analysis available in most biochemical laboratories, and it allows one to control the quality of the peptide on each bead, an important feature that is missing from most library decoding protocols. In the algorithm, each AA is assigned to two variable positions in the sequence grouped in a "unique pair". This arrangement limits sequence design because both the number of unique pairs U (setting the maximum number of variable AA) and the maximum number S of different AA per variable position depend on the peptide length N (U=N(N-1)/2), S=N-1). The method is therefore only suitable for focused libraries. An application example is shown for the selection of peptides with N-terminal proline or hydroxyproline catalyzing an aldol reaction from a combinatorial library of 65536 octapeptides. A simple enumeration program is available to help design combinatorial libraries decodable by amino acid analysis. The method applies to linear and cyclic peptides, can be used for nonnatural building blocks, including beta-amino acids, and should help to explore the vast chemistry of linear and cyclic peptide for catalysis and bioactivity. PMID- 17922555 TI - General microarray technique for immobilization and screening of natural glycans. AB - We here present a printed covalent glycan microarray for protein-binding studies, using low-femtomole quantities of glycans. Glycans, either natural glycans, which were released from glycoproteins and glycolipids from natural sources, or synthetic glycans, were labeled with common fluorescent labels (e.g., 2 aminobenzamide or 2-aminobenzoic acid) by reductive amination and purified by HPLC. The purified glycoconjugates were covalently immobilized on commercial epoxide-activated glass slides via the secondary amine group that links the glycan moiety with the fluorescent tag. This immobilization procedure is generally applicable to reductively aminated glycans with different established fluorescent labels and allows the spatial arrangement of oligosaccharides. The microarray comprised a variety of natural glycans from various biological sources and synthetic glycans and provided informative binding fingerprints for the lectin concanavalin A as well as 14 monoclonal antibodies. Recognized glycans were characterized by tandem mass spectrometry revealing binding motifs. This natural glycan array allowed the characterization of the specificity of carbohydrate-binding proteins for oligosaccharide ligands from sparse biological sources. Moreover, it was applied for the characterization of the microarray glycans by using known carbohydrate-binding proteins. PMID- 17922556 TI - Miniaturized total analysis systems: integration of electronics and fluidics using low-temperature co-fired ceramics. AB - The advantages of microanalyzers, usually fabricated in silicon, glass, or polymers, are well-known. The design and construction of fluidic platforms are well-developed areas due to the perfectly established microfabrication technologies used. However, there is still the need to achieve devices that include not only the fluid management system but also the measurement electronics, so that real portable miniaturized analyzers can be obtained. Low temperature co-fired ceramics technology permits the incorporation of actuators, such as micropumps and microvalves, controlled either magnetically, piezoelectrically, or thermally. Furthermore, electronic circuits can be also easily built exploiting the properties of these ceramics and the fact that they can be fabricated using a multilayer approach. In this work, taking advantage of the possibility of combining fluidics and electronics in a single substrate and using the same fabrication methodology, a chemical microanalyzer that integrates microfluidics, the detection system, and also the data acquisition and digital signal processing electronics is presented. To demonstrate the versatility of the technology, two alternative setups have been developed. In the first one, a modular configuration is proposed. In this case, the same electronic module can be used to determine different chemical parameters by simply exchanging the chemical module. In the second one, the monolithic integration of all the elements was accomplished, allowing the construction of compact and dedicated devices. Chloride ion microanalyzers have been constructed to demonstrate the operability of both device configurations. In all cases, the results obtained showed adequate analytical features. PMID- 17922557 TI - Application of analytical detection concepts to immunogenicity testing. AB - The cut point and detection limit of any immunogenicity assay are two of the most important quantities that define the adequacy of an assay for detecting anti-drug antibodies against therapeutic proteins. To date in the immunogenicity testing literature, only the type I (alpha) error (i.e., the false positive) rate of the assay has been considered for establishing cut points. The "sensitivity" of an immunogenicity assay is usually reported as the concentration of a monoclonal or polyclonal anti-drug antibody standard corresponding to the signal at the cut point. We propose that a more traditional and rigorous analytical chemistry definition of the detection capability be utilized wherein both type I and type II (beta, false negative) error rates are considered. Specifically, the Hubaux Vos technique of calculating cut points and limits of detection from predication intervals on calibration curves is recommended as a statistically rigorous approach. The utility of using receiver-operator characteristic curves for managing the type I and II error rates of an immunogenicity assay is also presented. In addition, we illustrate how a soluble receptor, sMUC18, for the therapeutic mAb ABX-MA1 can result in false positives by Biacore methodology. This result suggests that immunogenicity confirmatory experiments must be carefully designed, preferably with a smaller type I and II error rate than in the primary screening if an acceptable limit of detection can be maintained. PMID- 17922560 TI - A clinical pharmacokinetic analysis of tegafur-uracil (UFT) plus leucovorin given in a new twice-daily oral administration schedule. AB - BACKGROUND AND OBJECTIVE: Tegafur is an oral fluorouracil prodrug used in the treatment of colorectal cancer. The aim of this phase II, crossover, bioequivalence study was to compare the pharmacokinetics (primary objective) and tolerability (secondary objective) of tegafur-uracil (UFT) given as three daily doses (tid, reference schedule) with those obtained using a more convenient schedule of two daily doses (bid, new schedule). PATIENT AND METHODS: Twenty-one patients with metastatic colorectal cancer (median age 63 years) received the same oral daily dose of UFT (300 mg/m(2)/day) plus leucovorin (90 mg/day) divided into two or three daily doses. Patients were randomised to receive the first cycle either tid (12 patients) or bid (9 patients). The eligibility criteria included an Eastern Co-operative Oncology Group performance status of < or =1 and adequate bone-marrow, hepatic and renal function. The pharmacokinetics of uracil, fluorouracil and tegafur (high-performance liquid chromatography assays) were evaluated at steady state over 24 hours (area under the plasma concentration-time curve from 0 to 24 hours [AUC(24)], minimum plasma concentration [C(min)] and maximum plasma concentration [C(max)]). The pharmacokinetic parameters were analysed after logarithmic transformation according to a general linear model. RESULTS: The AUC(24)values of fluorouracil (p < 0.0001), uracil (p < 0.0001) and tegafur (p = 0.058) were greater with the bid schedule than the tid schedule. The bid : tid AUC(24) ratio (90% CI) was 1.8 (1.55, 2.10) with fluorouracil, 2.0 (1.59, 2.57) with uracil and 1.2 (1.02, 1.36) with tegafur, indicating that the bid and tid schedules were not bioequivalent. No major toxicity (grade 4) was reported, and grade 3 adverse events accounted for 9% of the total adverse events. Intra-patient comparison of the maximum toxicity grade did not demonstrate a significant difference between the bid and tid schedules (p = 0.18). CONCLUSION: A 2-fold increase in the fluorouracil and uracil AUC values was observed with UFT administered bid compared with tid, without a significant impact on tolerability, suggesting that the more convenient bid schedule may improve the UFT therapeutic index. PMID- 17922562 TI - How I became a lifelong learner. PMID- 17922559 TI - Effect of highly active antiretroviral therapy on tacrolimus pharmacokinetics in hepatitis C virus and HIV co-infected liver transplant recipients in the ANRS HC 08 study. AB - OBJECTIVE: To characterise the interactions between tacrolimus and antiretroviral drug combinations in hepatitis C virus-HIV co-infected patients who had received a liver transplant. DESIGN: An observational, open-label, multiple-dose, two period, one-sequence design clinical trial in which patients received tacrolimus as an immunosuppressive therapy during the postoperative period and then had an antiretroviral drug regimen added. Tacrolimus pharmacokinetics were evaluated at steady state during these two periods. METHODS: Fourteen patients participated in the study and seven participated in the intensified pharmacokinetic protocol. Patients were included if they had undergone liver transplantation for end-stage chronic hepatitis C, absence of opportunistic infection, a CD4 cell count of >150 cells/microL and an undetectable HIV plasma viral load (<50 copies/mL) under highly active antiretroviral therapy. During the posttransplantation period, the tacrolimus dose was adjusted according to blood concentrations. When liver function and the tacrolimus dose were stable, antiretroviral therapy was reintroduced. RESULTS: When lopinavir/ritonavir were added to the tacrolimus regimen (seven patients), the tacrolimus dose was reduced by 99% to maintain the tacrolimus concentration within the therapeutic range. Only two patients were treated with nelfinavir, which led to a wide variation in inhibition of tacrolimus metabolism. When efavirenz (four patients) or a nucleoside analogue combination (one patient) was added, very little change in tacrolimus dosing was required. CONCLUSION: The lopinavir/ritonavir combination markedly inhibited tacrolimus metabolism, whereas the effect of efavirenz was small. Tacrolimus dosing must be optimised according to therapeutic drug monitoring and the antiretroviral drug combination. PMID- 17922563 TI - Restless legs syndrome: nonpharmacologic and pharmacologic treatments. AB - Treatment of restless legs syndrome (RLS) has as its goals alleviation of the primary symptoms of the disorder and establishment of normal sleep. Dopamine agonists are considered first-line treatment when daily treatment for primary RLS is indicated. Gabapentin and opioids may be of value for refractory cases. Initial treatment of secondary RLS should address the underlying cause. PMID- 17922561 TI - Mechanism-based pharmacokinetic-pharmacodynamic modelling of the reversal of buprenorphine-induced respiratory depression by naloxone : a study in healthy volunteers. AB - BACKGROUND AND OBJECTIVE: Respiratory depression is a potentially life threatening adverse effect of opioid therapy. It has been postulated that the difficulty of reversing buprenorphine-induced respiratory depression is caused by slow receptor association-dissociation kinetics at the opioid mu receptor. The aim of this study was to characterise the pharmacodynamic interaction between buprenorphine and naloxone in healthy volunteers. METHODS: A competitive pharmacodynamic interaction model was proposed to describe and predict the time course of naloxone-induced reversal of respiratory depression. The model was identified using data from an adaptive naloxone dose-selection trial following intravenous administration of buprenorphine 0.2mg/70kg or 0.4mg/70kg. RESULTS: The pharmacokinetics of naloxone and buprenorphine were best described by a two compartment model and a three-compartment model, respectively. A combined biophase equilibration-receptor association-dissociation pharmacodynamic model described the competitive interaction between buprenorphine and naloxone at the opioid mu receptor. For buprenorphine, the values of the rate constants of receptor association (k(on)) and dissociation (k(off)) were 0.203 mL/ng/min and 0.0172 min(-)(1), respectively. The value of the equilibrium dissociation constant (K(D)) was 0.18 nmol/L. The half-life (t((1/2))) of biophase equilibration was 173 minutes. These estimates of the pharmacodynamic parameters are similar to values obtained in the absence of naloxone co-administration. For naloxone, the half-life of biophase distribution was 6.5 minutes. CONCLUSIONS: Because of the slow receptor association-dissociation kinetics of buprenorphine in combination with the fast elimination kinetics of naloxone, naloxone is best administered as a continuous infusion for reversal of buprenorphine-induced respiratory depression. PMID- 17922564 TI - When are indwelling urinary catheters appropriate in elderly patients? AB - Indwelling catheters are commonly used in elderly patients with urinary retention, incontinence, pressure ulcers, and cancer, ostensibly for the patient's comfort but sometimes to ease the burden of health care workers. Indwelling catheterization is overused; catheters should be inserted only for specific, well-documented indications. The best way to avoid catheter-associated infections and other complications is to avoid long-term catheter use whenever possible. Given the risks involved, catheterization is indicated only as a last resort, after other options have failed. PMID- 17922558 TI - Pharmacokinetic considerations in clinical toxicology: clinical applications. AB - Pharmacokinetic and pharmacodynamic principles should be regarded in the assessment and proper management of patients exposed to a poison. Clinicians must apply these principles to make rational clinical decisions regarding the significance of the poisoning (risk assessment) and to formulate an appropriate management plan. However, pharmacokinetic processes and parameters may be changed in the patient with acute poisoning. This may result from saturation of the capacity of a number of physiological processes due to the high dose, or the toxic effects of the poison may change these processes directly. For example, absorption kinetics may be altered because of increased gastrointestinal transit time (e.g. cholinergic receptor antagonists) or saturable absorption (e.g. methotrexate). Saturation of protein binding may increase the volume of distribution and thereby increase the elimination half-life (e.g. salicylates). Alteration of the acid-base balance (poison-induced or iatrogenic) may also increase or decrease the distribution of a poison. Saturation of metabolism at high doses can prolong toxicity (e.g. phenytoin) or lead to other routes of metabolism that lead to increased toxicity (e.g. paracetamol [acetaminophen]). Excretion may be reduced by saturation of active transporters or decreased renal blood flow.A better understanding of pharmacokinetic principles should improve the clinical care of patients. It should lead to more accurate interpretation of blood concentrations or biomarkers (e.g. ECG intervals or acetylcholinesterase activity) and how these relate to the time course for that poison, and better prediction of prognosis. This in turn, indicates the appropriate duration of observation and the requirement for some specific treatments. Many specific poisoning treatments aim to favourably alter the pharmacokinetics of the poison. These include activated charcoal, whole bowel irrigation, extracorporeal elimination, chelating agents, antitoxins and urinary alkalinisation. The evidence supporting them, their indications and limitations can only be understood using pharmacokinetic principles. These principles also underpin the appropriate choice within the flexible dosage regimen for many antidotes. In particular, naloxone, flumazenil, methylene blue, atropine and pralidoxime all use variable doses and have an elimination half-life that is much shorter than many (but not all) of the poisons treated by these agents. A firm grounding in pharmacokinetics/toxicokinetics should be regarded as a core competency for all professionals involved in clinical care or undertaking research in clinical toxicology. PMID- 17922565 TI - Nutrition in the elderly: diet pitfalls and nutrition advice. AB - This final installment of a special series on nutrition in the elderly considers dietary pitfalls and their sequelae. Years of poor dietary habits contribute to biological risk and lifestyle changes in the elderly. Clinicians must properly evaluate the nutritional status of their older patients to restore nutritional adequacy and healthy aging. PMID- 17922566 TI - Blistering eruption involving most of the body. PMID- 17922567 TI - Heat shock response in CHO mammalian cells is controlled by a nonlinear stochastic process. AB - In many biological systems, the interactions that describe the coupling between different units in a genetic network are nonlinear and stochastic. We study the interplay between stochasticity and nonlinearity using the responses of Chinese hamster ovary (CHO) mammalian cells to different temperature shocks. The experimental data show that the mean value response of a cell population can be described by a mathematical expression (empirical law) which is valid for a large range of heat shock conditions. A nonlinear stochastic theoretical model was developed that explains the empirical law for the mean response. Moreover, the theoretical model predicts a specific biological probability distribution of responses for a cell population. The prediction was experimentally confirmed by measurements at the single-cell level. The computational approach can be used to study other nonlinear stochastic biological phenomena. PMID- 17922568 TI - Universally sloppy parameter sensitivities in systems biology models. AB - Quantitative computational models play an increasingly important role in modern biology. Such models typically involve many free parameters, and assigning their values is often a substantial obstacle to model development. Directly measuring in vivo biochemical parameters is difficult, and collectively fitting them to other experimental data often yields large parameter uncertainties. Nevertheless, in earlier work we showed in a growth-factor-signaling model that collective fitting could yield well-constrained predictions, even when it left individual parameters very poorly constrained. We also showed that the model had a "sloppy" spectrum of parameter sensitivities, with eigenvalues roughly evenly distributed over many decades. Here we use a collection of models from the literature to test whether such sloppy spectra are common in systems biology. Strikingly, we find that every model we examine has a sloppy spectrum of sensitivities. We also test several consequences of this sloppiness for building predictive models. In particular, sloppiness suggests that collective fits to even large amounts of ideal time-series data will often leave many parameters poorly constrained. Tests over our model collection are consistent with this suggestion. This difficulty with collective fits may seem to argue for direct parameter measurements, but sloppiness also implies that such measurements must be formidably precise and complete to usefully constrain many model predictions. We confirm this implication in our growth-factor-signaling model. Our results suggest that sloppy sensitivity spectra are universal in systems biology models. The prevalence of sloppiness highlights the power of collective fits and suggests that modelers should focus on predictions rather than on parameters. PMID- 17922569 TI - Metabolic reconstruction and modeling of nitrogen fixation in Rhizobium etli. AB - Rhizobiaceas are bacteria that fix nitrogen during symbiosis with plants. This symbiotic relationship is crucial for the nitrogen cycle, and understanding symbiotic mechanisms is a scientific challenge with direct applications in agronomy and plant development. Rhizobium etli is a bacteria which provides legumes with ammonia (among other chemical compounds), thereby stimulating plant growth. A genome-scale approach, integrating the biochemical information available for R. etli, constitutes an important step toward understanding the symbiotic relationship and its possible improvement. In this work we present a genome-scale metabolic reconstruction (iOR363) for R. etli CFN42, which includes 387 metabolic and transport reactions across 26 metabolic pathways. This model was used to analyze the physiological capabilities of R. etli during stages of nitrogen fixation. To study the physiological capacities in silico, an objective function was formulated to simulate symbiotic nitrogen fixation. Flux balance analysis (FBA) was performed, and the predicted active metabolic pathways agreed qualitatively with experimental observations. In addition, predictions for the effects of gene deletions during nitrogen fixation in Rhizobia in silico also agreed with reported experimental data. Overall, we present some evidence supporting that FBA of the reconstructed metabolic network for R. etli provides results that are in agreement with physiological observations. Thus, as for other organisms, the reconstructed genome-scale metabolic network provides an important framework which allows us to compare model predictions with experimental measurements and eventually generate hypotheses on ways to improve nitrogen fixation. PMID- 17922570 TI - Growth of H5N1 influenza A viruses in the upper respiratory tracts of mice. AB - Highly pathogenic avian H5N1 influenza A viruses have spread throughout Asia, Europe, and Africa, raising serious worldwide concern about their pandemic potential. Although more than 250 people have been infected with these viruses, with a consequent high rate of mortality, the molecular mechanisms responsible for the efficient transmission of H5N1 viruses among humans remain elusive. We used a mouse model to examine the role of the amino acid at position 627 of the PB2 viral protein in efficient replication of H5N1 viruses in the mammalian respiratory tract. Viruses possessing Lys at position 627 of PB2 replicated efficiently in lungs and nasal turbinates, as well as in cells, even at the lower temperature of 33 degrees C. Those viruses possessing Glu at this position replicated less well in nasal turbinates than in lungs, and less well in cells at the lower temperature. These results suggest that Lys at PB2-627 confers to avian H5N1 viruses the advantage of efficient growth in the upper and lower respiratory tracts of mammals. Therefore, efficient viral growth in the upper respiratory tract may provide a platform for the adaptation of avian H5N1 influenza viruses to humans and for efficient person-to-person virus transmission, in the context of changes in other viral properties including specificity for human (sialic acid alpha-2,6-galactose containing) receptors. PMID- 17922571 TI - A single mutation in the PB1-F2 of H5N1 (HK/97) and 1918 influenza A viruses contributes to increased virulence. AB - The proapoptotic PB1-F2 protein of influenza A viruses has been shown to contribute to pathogenesis in the mouse model. Expression of full-length PB1-F2 increases the pathogenesis of the influenza A virus, causing weight loss, slower viral clearance, and increased viral titers in the lungs. After comparing viruses from the Hong Kong 1997 H5N1 outbreak, one amino acid change (N66S) was found in the PB1-F2 sequence at position 66 that correlated with pathogenicity. This same amino acid change (N66S) was also found in the PB1-F2 protein of the 1918 pandemic A/Brevig Mission/18 virus. Two isogenic recombinant chimeric viruses were created with an influenza A/WSN/33 virus background containing the PB1 segment from the HK/156/97: WH and WH N66S. In mice infected with WH N66S virus there was increased pathogenicity as measured by weight loss and decreased survival, and a 100-fold increase in virus replication when compared to mice infected with the WH virus. The 1918 pandemic strain A/Brevig Mission/18 was reconstructed with a pathogenicity-reducing mutation in PB1-F2 (S66N). The resultant 1918 S66N virus was attenuated in mice having a 3-log lower 50% lethal dose and caused less morbidity and mortality in mice than the wild-type virus. Viral lung titers were also decreased in 1918 S66N-infected mice compared with wild-type 1918 virus-infected mice. In addition, both viruses with an S at position 66 (WH N66S and wt 1918) induced elevated levels of cytokines in the lungs of infected mice. Together, these data show that a single amino acid substitution in PB1-F2 can result in increased viral pathogenicity and could be one of the factors contributing to the high lethality seen with the 1918 pandemic virus. PMID- 17922572 TI - A viral nanoparticle with dual function as an anthrax antitoxin and vaccine. AB - The recent use of Bacillus anthracis as a bioweapon has stimulated the search for novel antitoxins and vaccines that act rapidly and with minimal adverse effects. B. anthracis produces an AB-type toxin composed of the receptor-binding moiety protective antigen (PA) and the enzymatic moieties edema factor and lethal factor. PA is a key target for both antitoxin and vaccine development. We used the icosahedral insect virus Flock House virus as a platform to display 180 copies of the high affinity, PA-binding von Willebrand A domain of the ANTXR2 cellular receptor. The chimeric virus-like particles (VLPs) correctly displayed the receptor von Willebrand A domain on their surface and inhibited lethal toxin action in in vitro and in vivo models of anthrax intoxication. Moreover, VLPs complexed with PA elicited a potent toxin-neutralizing antibody response that protected rats from anthrax lethal toxin challenge after a single immunization without adjuvant. This recombinant VLP platform represents a novel and highly effective, dually-acting reagent for treatment and protection against anthrax. PMID- 17922573 TI - Ancient exaptation of a CORE-SINE retroposon into a highly conserved mammalian neuronal enhancer of the proopiomelanocortin gene. AB - The proopiomelanocortin gene (POMC) is expressed in the pituitary gland and the ventral hypothalamus of all jawed vertebrates, producing several bioactive peptides that function as peripheral hormones or central neuropeptides, respectively. We have recently determined that mouse and human POMC expression in the hypothalamus is conferred by the action of two 5' distal and unrelated enhancers, nPE1 and nPE2. To investigate the evolutionary origin of the neuronal enhancer nPE2, we searched available vertebrate genome databases and determined that nPE2 is a highly conserved element in placentals, marsupials, and monotremes, whereas it is absent in nonmammalian vertebrates. Following an in silico paleogenomic strategy based on genome-wide searches for paralog sequences, we discovered that opossum and wallaby nPE2 sequences are highly similar to members of the superfamily of CORE-short interspersed nucleotide element (SINE) retroposons, in particular to MAR1 retroposons that are widely present in marsupial genomes. Thus, the neuronal enhancer nPE2 originated from the exaptation of a CORE-SINE retroposon in the lineage leading to mammals and remained under purifying selection in all mammalian orders for the last 170 million years. Expression studies performed in transgenic mice showed that two nonadjacent nPE2 subregions are essential to drive reporter gene expression into POMC hypothalamic neurons, providing the first functional example of an exapted enhancer derived from an ancient CORE-SINE retroposon. In addition, we found that this CORE-SINE family of retroposons is likely to still be active in American and Australian marsupial genomes and that several highly conserved exonic, intronic and intergenic sequences in the human genome originated from the exaptation of CORE-SINE retroposons. Together, our results provide clear evidence of the functional novelties that transposed elements contributed to their host genomes throughout evolution. PMID- 17922576 TI - American Society of Clinical Oncology/College of American Pathologists guidelines should be scientifically validated. PMID- 17922574 TI - Power to detect risk alleles using genome-wide tag SNP panels. AB - Advances in high-throughput genotyping and the International HapMap Project have enabled association studies at the whole-genome level. We have constructed whole genome genotyping panels of over 550,000 (HumanHap550) and 650,000 (HumanHap650Y) SNP loci by choosing tag SNPs from all populations genotyped by the International HapMap Project. These panels also contain additional SNP content in regions that have historically been overrepresented in diseases, such as nonsynonymous sites, the MHC region, copy number variant regions and mitochondrial DNA. We estimate that the tag SNP loci in these panels cover the majority of all common variation in the genome as measured by coverage of both all common HapMap SNPs and an independent set of SNPs derived from complete resequencing of genes obtained from SeattleSNPs. We also estimate that, given a sample size of 1,000 cases and 1,000 controls, these panels have the power to detect single disease loci of moderate risk (lambda approximately 1.8-2.0). Relative risks as low as lambda approximately 1.1-1.3 can be detected using 10,000 cases and 10,000 controls depending on the sample population and disease model. If multiple loci are involved, the power increases significantly to detect at least one locus such that relative risks 20%-35% lower can be detected with 80% power if between two and four independent loci are involved. Although our SNP selection was based on HapMap data, which is a subset of all common SNPs, these panels effectively capture the majority of all common variation and provide high power to detect risk alleles that are not represented in the HapMap data. PMID- 17922578 TI - Malakoplakia outside the urinary tract. PMID- 17922579 TI - Malakoplakia outside the urinary tract. PMID- 17922581 TI - Relevance of the autopsy as a medical tool: a large database of physician attitudes. PMID- 17922583 TI - Cutaneous pulse granulomas. PMID- 17922585 TI - Emergency transfusion of incompatible red blood cells. PMID- 17922586 TI - The role of gadolinium in triggering nephrogenic systemic fibrosis/nephrogenic fibrosing dermopathy. PMID- 17922587 TI - Best practices in diagnostic immunohistochemistry: pleomorphic cutaneous spindle cell tumors. AB - CONTEXT: Pleomorphic cutaneous spindle cell tumors can be difficult to distinguish solely on histologic grounds. The use of ancillary immunohistochemical studies can greatly assist in this differential diagnosis. OBJECTIVE: To review histologic and immunohistochemical aspects of cutaneous spindle cell tumors and discuss a basic panel of markers to assist in the differential diagnosis. DATA SOURCES: English-language literature published between 1981 and 2005. CONCLUSIONS: A basic immunohistochemistry panel for high molecular-weight cytokeratin, melanocytic markers (S100 protein, HMB-45, Melan A), smooth muscle actin, desmin, and endothelial markers (CD31, CD34) is effective in diagnosing most cutaneous spindle cell tumors. PMID- 17922588 TI - The value of monitoring human papillomavirus DNA results for Papanicolaou tests diagnosed as atypical squamous cells of undetermined significance: a College of American Pathologists Q-Probes study of 68 institutions. AB - CONTEXT: Papanicolaou (Pap) tests are often diagnosed as atypical squamous cells of undetermined significance (ASC-US). Human papillomavirus (HPV) DNA testing has been proposed as a quality metric for this diagnosis. OBJECTIVE: To measure the frequency of HPV positivity in Pap tests diagnosed as ASC-US and to examine laboratory variables that are associated with institutional deviation from the mean percent of HPV positivity. DESIGN: As part of a College of American Pathologist Q-Probes program, 68 participating laboratories retrospectively identified approximately 50 consecutive ASC-US Pap tests that had HPV testing results. RESULTS: The mean percentage of HPV positivity for ASC-US was 43.74% among institutions surveyed, but it had a broad distribution, with an SD of 17.77%. Associations were found for lower difference of the institutional mean from the surveyed interinstitutional mean percentage of positive HPV with (1) higher numbers of Pap tests in the past year that had HPV testing, (2) in-house HPV testing, and (3) teaching hospitals. All 3 factors correlated with a larger volume of Pap tests per institution. An association was found between patient age and the probability of a positive HPV result, indicating a dependence upon prevalence of HPV. CONCLUSIONS: Larger volumes of Pap tests may offer an opportunity to gain greater comfort in interpreting Pap tests. While there is significant variability in interinstitutional HPV-positive rates in ASC-US Pap tests, monitoring the HPV-positive rate in ASC-US Pap tests is a valuable broad measure of quality. Performance beyond 2 SDs of the mean should prompt reassessment of diagnostic criteria used in the evaluation of Pap tests and/or investigation of the prevalence of HPV positivity in the population from which the Pap tests are obtained. PMID- 17922589 TI - Frozen section discrepancy in the evaluation of central nervous system tumors. AB - CONTEXT: Frozen section (FS) evaluation of central nervous system (CNS) lesions provides an assessment of specimen adequacy and facilitates triage for ancillary studies. Frozen section also provides an accurate preliminary diagnosis; however, certain lesions are recognized to cause diagnostic challenges at FS. OBJECTIVE: To identify cases in which there was a discrepancy between the FS diagnosis and final diagnosis in the case to heighten awareness of common diagnostic pitfalls in surgical neuropathology. DESIGN: All CNS FS cases involving a tumor diagnosis at FS or permanent section (N = 2156) from September 1997 until June 2005 were retrospectively reviewed. Discrepancies between the FS and final diagnoses were identified. RESULTS: Of the 2156 cases identified, 57 (2.7%) discrepant diagnoses were found. Twelve (21.1%) of 57 discrepancies involved errors in classification of spindle cell lesions, most commonly confusing schwannomas or meningiomas with other lesions. Twelve (21.1%) of 57 cases involved errors in differentiating oligodendrogliomas from astrocytomas. Nine (15.8%) of 57 discrepancies involved errors in the diagnosis of CNS lymphoma. Eight (14.0%) of 57 cases involved errors in differentiating reactive from neoplastic processes, most frequently gliosis versus glioma. Four (7.0%) of 57 discrepancies involved errors in the overgrading of tumors. The remaining 12 (21.1%) of 57 cases included an assortment of other discrepancies. CONCLUSIONS: Frozen section of CNS neoplastic processes can be highly accurate. Less than 3% of FS diagnoses in 1 institution's experience were discrepant with the final diagnoses. Approximately 80% of the discrepant cases were classified into 5 categories: spindle cell lesions, astrocytoma versus oligodendroglioma, differential diagnosis of CNS lymphoma, reactive versus neoplastic process, and tumor overgrading. Awareness of these pitfalls may help in further increasing diagnostic accuracy. PMID- 17922590 TI - Claudin-7 immunohistochemistry in renal tumors: a candidate marker for chromophobe renal cell carcinoma identified by gene expression profiling. AB - CONTEXT: The differential diagnosis of eosinophilic renal tumors can be difficult by light microscopy. In particular, chromophobe renal cell carcinoma (RCC) is difficult to distinguish from oncocytoma. This differential diagnosis is important because chromophobe RCC is malignant, whereas oncocytoma is benign. Furthermore, chromophobe RCC has distinct malignant potential and prognosis compared with eosinophilic variants of other RCC subtypes. Immunohistochemistry is useful for distinguishing chromophobe RCC from other subtypes of renal carcinoma, but no expression marker reliably separates chromophobe RCC from oncocytoma. OBJECTIVE: In a previous gene expression microarray analysis of renal tumor subtypes, we found the distal nephron markers claudin-7 and claudin-8 to be overexpressed in chromophobe RCC versus oncocytoma and other tumor subtypes. We have confirmed similar findings in independent microarray data and validated differential claudin-7 protein expression by immunohistochemistry. DESIGN: Immunohistochemical analysis of claudin-7 in 36 chromophobe RCCs, 43 oncocytomas, 42 clear cell RCCs, and 29 papillary RCCs. RESULTS: Membranous claudin-7 expression was detected in 67% chromophobe RCCs, compared with 0% clear cell RCCs, 28% papillary RCCs, and 26% oncocytomas (P < .001). CONCLUSIONS: Based on microarray and immunohistochemical data, we propose claudin-7 to be a candidate expression marker for distinguishing chromophobe RCC from other renal tumor subtypes, including the morphologically similar oncocytoma. The clinical utility of claudin-7 should be validated in independent studies of renal tumors, possibly in combination with additional targets in a multiplex immunohistochemical panel. PMID- 17922591 TI - Critical leadership and management skills for pathology practice. AB - CONTEXT: Managed care has changed the nature of medical practice. The practice of pathology has also changed and is likely to undergo further modification. Additional skills in leadership and management are needed to perform optimally in the current medical marketplace. OBJECTIVE: To determine whether pathologists view business and informatics skills as being important and valuable in their practices. DESIGN: A survey was sent electronically (via e-mail) to 2566 pathologists and pathology administrators. RESULTS: Two hundred sixteen survey recipients (8.4%) responded to the survey. The response rate to individual questions ranged from 86% to 99% (186 to 214 of 216 total respondents). Most who took the survey ranked communication (203/209; 97%), leadership skills (165/209; 79%), and systems thinking skills (155/209; 74%) as either "very important" or "essential." Fewer respondents were willing to offer salary premiums for marketing (108/196; 55%), business or finance (92/196; 47%), or staff leadership (95/196; 48%) skills unless the candidate had a track record of measurable achievement using these skills. Those practicing in academia as well as those making hiring decisions by themselves were more likely to value leadership and management skills. Fewer than 1% of respondents in any category considered current pathology residency training in leadership and management to be adequate. CONCLUSIONS: Prospective employers value leadership and management skills. Pathology residency programs must include meaningful training in pathology practice management and informatics to satisfy the demand for these skills in the modern pathology marketplace. PMID- 17922592 TI - Alpha-methylacyl CoA racemase in pulmonary adenocarcinoma, squamous cell carcinoma, and neuroendocrine tumors: expression and survival analysis. AB - CONTEXT: Alpha-methylacyl CoA racemase (AMACR) is an oxidative enzyme involved in isomeric transformation of fatty acids entering the beta-oxidation pathway. AMACR serves as a useful marker in establishing a diagnosis of prostatic malignancy; however, limited information is available in regard to its presence in pulmonary neoplasms. OBJECTIVE: To investigate AMACR expression within a spectrum of lung carcinomas and its correlation with patients' survival. DESIGN: Four hundred seventy-seven pulmonary carcinomas, including 150 squamous cell carcinomas, 150 adenocarcinomas, 46 typical carcinoids, 31 atypical carcinoids, 28 large cell neuroendocrine carcinomas, and 72 small cell carcinomas, were studied immunohistochemically using tissue microarray-based samples. RESULTS: Overall, pulmonary tumors were positive for AMACR in a significant percentage (47%) of cases. Among tumor types, 22% of squamous cell carcinoma, 56% of adenocarcinoma, 72% of typical carcinoid, 52% of atypical carcinoid, 70% of large cell neuroendocrine carcinoma, and 51% of small cell lung carcinoma were positive for AMACR. Furthermore, the Kaplan-Meier analysis revealed that the patients with AMACR-positive small cell carcinoma had better survival (19% vs 5% after 5 years, P = .04) than patients with AMACR-negative tumors. Such survival advantage was seen for patients with stage I-II (P = .01) but not stage III-IV small cell carcinomas (P = .58). CONCLUSIONS: These results indicate that, similar to prostate cancer, the overexpression of AMACR frequently occurs in pulmonary carcinomas. Additionally, its positive correlation with outcome of stage I-II small cell lung carcinoma warrants further investigation of the AMACR role in the prognosis of this tumor. PMID- 17922593 TI - Panels of immunohistochemical markers help determine primary sites of metastatic adenocarcinoma. AB - CONTEXT: Although identification of the primary tumor in patients with metastatic adenocarcinoma has a profound clinical impact, diagnosing the organ of origin is frequently difficult. Because none of the individual immunohistochemical markers used for tissue identification are both site specific and site sensitive, multiple markers are needed to improve the prediction of primary sites. OBJECTIVE: To develop an effective approach to immunohistochemically evaluate metastatic adenocarcinoma for the assignment of a likely primary site of origin. DESIGN: Expression profiles of CDX2, cytokeratin (CK) 7, CK20, thyroid transcription factor 1 (TTF-1), carcinoembryonic antigen (CEA), MUC2, MUC5AC, SMAD4, estrogen receptor (ER), and gross cystic disease fluid protein 15 (GCDFP 15) were generated in adenocarcinomas from 7 primary sites, followed by construction of a decision tree and design of multiple-marker panels. Expression of these markers was evaluated immunohistochemically in 314 primary adenocarcinomas (50 cases each of colorectal, gastric, lung, pancreatic, bile duct, and breast, and 14 cases of ovarian origin) using the tissue array method. Results were validated using 60 cases of metastatic adenocarcinoma with known primaries. RESULTS: Organ-specific immunostaining profiles using multiple markers provided high sensitivity, specificity, and positive predictive value in detecting primary adenocarcinomas, as follows: colorectal, TTF-1-/CDX2+/CK7 /CK20+ or TTF-1-/CDX2+/CK7-/CK20-/(CEA+ or MUC2+); ovarian, CK7+/MUC5AC+/TTF-1 /CDX2-/CEA-/GCDFP-15-; breast, GCDFP-15+/TTF-1-/CDX2-/CK7+/CK20- or ER+/ TTF-1 /CDX2-/CK20-/CEA-/MUC5AC-; lung, TTF-1+ or TTF-1-/CDX2-/CK7+/CK20-/GCDFP-15-/ER /CEA-/ MUC5AC-; pancreaticobiliary, TTF-1-/CDX2-/CK7+/ CEA+/MUC5AC+; and stomach, TTF-1-/CDX2+/CK7+/ CK20-. Overall, these combined phenotypes correctly predicted the tester samples (metastatic adenocarcinomas with known primaries) in 75% of cases. CONCLUSIONS: Determination of tissue-specific immunostaining profiles is valuable in the diagnostic differentiation of metastatic adenocarcinomas from seven common primary sites and should help to correctly predict the organ of primary tumor origin. PMID- 17922594 TI - Quantification of the morphologic features of fibroepithelial tumors of the breast. AB - CONTEXT: Phyllodes tumors of the breast are uncommon, comprising 0.3% to 0.9% of female primary breast tumors. Owing in part to their rarity, definitive, objective, reproducible morphologic criteria that reliably distinguish benign from low-grade malignant or malignant phyllodes tumors have yet to be established. OBJECTIVE: To use image analysis to quantitate and compare morphologic features of different groups of fibroepithelial tumors (FETs) of the breast. DESIGN: Hematoxylin-eosin-stained sections of 41 FETs previously identified as fibroadenoma, benign phyllodes, low-grade malignant phyllodes, or high-grade malignant phyllodes were blinded and studied using a Leica DMRA2 microscope and OpenLab Image Analysis software. Features measured included mitotic rate per 10 high-power fields, stromal cellularity, nuclear size, stromal overgrowth, and the largest and smallest stromal-epithelial surface area ratios. Epithelial appearance was measured on a semiquantitative basis. Features of each case including tumor size, margin status, and the presence of necrosis or heterologous elements were also considered; these data were retrieved from surgical pathology reports. RESULTS: Quantitative measures of stromal cellularity, stromal-epithelial ratio, mitotic rate, stromal overgrowth, and mean nuclear diameter were developed and found to stratify a population of FETs by the current classification system of fibroadenoma, benign, and low-grade or high grade malignant phyllodes tumor. CONCLUSIONS: Quantitative morphologic features of FETs can be used to stratify these tumors by subtype. Use of these quantitative criteria could reduce interrater variability in histologically identifying FETs by subclass. PMID- 17922595 TI - Improved filter method for urine sediment detection of urothelial carcinoma by fluorescence in situ hybridization. AB - CONTEXT: Fluorescence in situ hybridization (FISH) of voided urine sediment is a sensitive and specific test for the detection of urothelial carcinoma. The time required for slide preparation using the conventional cytospin method is lengthy. OBJECTIVE: To present an alternative to the conventional cytospin method. DESIGN: We compared the results of an improved filter monolayer method with published results of the conventional cytospin method. A total of 624 patients with cytology and FISH analyses were followed with cystoscopy and/or bladder biopsy. Fluorescence in situ hybridization analysis was performed on 624 cases using fluorescence-labeled probes to the pericentromeric regions of chromosomes 3, 7, and 17 and band 9p21; cytology was also performed in all cases. RESULTS: A total of 217 (34.7%) of 624 patients had follow-up bladder biopsies, and 170 of these (78.3%) had urothelial carcinoma. The sensitivity for cancer detection was higher for FISH than for urine cytology (92.9% [158/ 170] for FISH vs 72.9% [124/170] for urine cytology, P = <5%). The specificity was equivalent for FISH and urine cytology (97.5% [443/454] for FISH vs 92.2% [419/454] for cytology). The sensitivity for FISH was better (92.9% vs 81%), and there was no significant difference in specificity (97.5% vs 96%) between the filter method and the conventional cytospin method. Unlike the conventional cytospin method, the filter method did not require multiple centrifugation and decantation steps or investment in dedicated equipment. CONCLUSIONS: The improved filter method was faster, easier, and less expensive than published results with the conventional cytospin method with better sensitivity and equivalent specificity. PMID- 17922596 TI - Omentectomy for gynecologic cancer: how much sampling is adequate for microscopic examination? AB - CONTEXT: Detecting omental metastasis is crucial for staging and treatment of endometrial and ovarian carcinoma. OBJECTIVE: To determine the optimal omental sampling for omentectomies to ascertain the stage of the disease in a cost effective way. DESIGN: We reevaluated 258 omentectomies that were performed due to ovarian or endometrial carcinoma. A total of 116 cases were retrospectively studied, and 142 cases were prospectively studied. For prospective study, 10 to 16 blocks were sampled if the omentum showed no signs of gross tumor. Mean omental block sample frequency of 2 groups with the negative macroscopy but with or without microscopic tumor have been compared using an independent samples t test. RESULTS: Seven patients had no evidence of tumor metastasis on gross examination but had microscopic tumor metastasis. The mean numbers of blocks were 6.4 for patients having microscopic tumor without macroscopic involvement and 7.8 for patients having neither microscopic nor macroscopic involvement. Approximately twice as many samples were taken in the prospective analysis when compared with retrospective analysis. Two cases with microscopic omental metastasis that had no macroscopic involvement at first impression were reevaluated retrospectively and found to contain 0.3- to 0.5-cm white nodules. The rate of omental metastasis increased with the grade of the tumor (P = .005). CONCLUSION: Careful macroscopic examination is the most important step in detecting small omental metastasis. For cases with gross tumor, one section is sufficient. If a macroscopic lesion is not detectable and the patient has a high grade tumor that will necessitate an adjuvant therapy, 3 to 5 samples seem sufficient for staging. Further studies are needed to determine the optimum sample size for tumors having a low risk of metastasis. PMID- 17922597 TI - Yield of primary and repeat induced sputum testing for Pneumocystis jiroveci in human immunodeficiency virus-positive and -negative patients. AB - CONTEXT: Induced sputum sampling has an approximate 70% sensitivity for detection of Pneumocystis jiroveci in human immunodeficiency virus (HIV) patients. Bronchoalveolar lavage sampling has greater than 90% sensitivity but is a far more invasive procedure. Therefore, bronchoalveolar lavage testing is often recommended as a follow-up after a negative induced sputum. In HIV-negative patients, the utility of induced sputum testing is still not well defined. OBJECTIVE: To determine whether repeat induced sputum sampling increases diagnostic yield and might thereby reduce the need for follow-up bronchoalveolar lavage sampling. To determine the utility of induced sputum sampling in HIV negative patients. DESIGN: A 2-year retrospective review of the utility of repeat induced sputa testing in patients with previous first and/or second negative induced sputa. Retrospective review of induced sputa detection in HIV-negative patients. RESULTS: Repeat testing of induced sputa for Pneumocystic jirovecii did not significantly increase diagnostic yield. Furthermore, in HIV-negative patients, induced sputum testing was diagnostically insensitive. CONCLUSIONS: Bronchoalveolar lavage testing should be performed initially in HIV-negative patients and after a first negative induced sputum in HIV-positive patients. PMID- 17922598 TI - Second messenger systems in human gliomas. AB - CONTEXT: Patients with glioblastoma (astrocytoma, World Health Organization grade IV) exhibit 2-year survival rates of less than 20% despite significant advances in therapeutic options available to patients. Epidermal growth factor receptor (EGFR) hyperexpression is one of the most commonly encountered abnormalities in this tumor. However, EGFR expression, amplification, and mutations are poorly predictive of patient survival. Investigators have taken to exploiting the sensitivities of activated downstream targets in the EGFR second messenger pathways to certain inhibitory drugs to downregulate their neoplastic messages promoting cell growth and inhibiting cell death. OBJECTIVE: It is important to both gain some understanding of the functional significance of these pathways and to understand the role the pathologist might play in characterizing the activation status of certain downstream messenger proteins that are targeted in these brain tumor therapies. We have reviewed the literature regarding histologic assays that have been incorporated into trials of these new drugs and report on the methods used to study these proteins and the conclusions of these studies. DATA SOURCES: Literature review and primary material from Duke University (Durham, NC) Department of Pathology archives. CONCLUSIONS: To date, drug trial reports indicate that identification of the presence of the EGFR variant, EGFRvIII, and measurement of the activated downstream targets, phospho-Akt, phospho-S6, and phospho-MAPK, may be useful in predicting sensitivity to some of the EGFR kinase inhibitors. No studies to date have identified prognostic significance related to immunoreactivity status among any of these markers that is independent of histologic grade. PMID- 17922599 TI - A case of fatal disseminated Bartonella henselae infection (cat-scratch disease) with encephalitis. AB - Cat-scratch disease resulting from Bartonella henselae infection is usually a benign, self-limited process in immunocompetent children. Even the rare cases associated with neurologic manifestations are not generally fatal. We report a case of a previously healthy 6-year-old boy with cat-scratch disease, systemic dissemination, and encephalitis that led to his death. Autopsy revealed perivascular lymphocytic infiltrates and microglial nodules in the brain. To our knowledge, this finding has not been previously reported in B. henselae infection, possibly because of the paucity of material available for complete neuropathologic evaluation. This case illustrates the extreme severity of the spectrum with which cat-scratch disease can present and provides evidence of brain histopathology that may be representative of the disease. PMID- 17922600 TI - Canalicular adenoma arising in the esophagus. AB - Canalicular adenomas are benign neoplasms that arise from salivary glands and often present as painless enlarging nodules. They have a predilection for upper lip but can be found throughout the oropharynx. To our knowledge, canalicular adenoma arising in the esophagus has never been described in the English literature. Here we report a canalicular adenoma occurring in the esophagus. PMID- 17922601 TI - Eosinophilic pustular folliculitis. AB - Classical eosinophilic pustular folliculitis, or Ofuji's disease, is a chronic and relapsing dermatosis that is predominantly reported in East Asian populations. Clinically, the disease typically begins as small papules, which enlarge and coalesce into a large plaque, usually on the face. The histopathology is characterized by a prominent eosinophilic infiltrate in the dermis with concentration around pilosebaceous units, often with eosinophilic microabscess formation. The differentiation of eosinophilic pustular folliculitis from other eosinophilic dermatoses is practically challenging and requires close clinicopathologic correlation. Eosinophilic pustular folliculitis may also be associated with human immunodeficiency virus infection, various drugs, and some lymphomas and could also be thought of as a nonspecific dermatopathologic pattern in such settings. The cause of classical eosinophilic pustular folliculitis is unknown, although immune processes are almost certain to play a key role in its pathogenesis. PMID- 17922602 TI - Basal cell adenocarcinoma of minor salivary glands. AB - Basal cell adenocarcinoma of minor salivary glands is a relatively rare slow growing tumor with an infiltrating growth pattern. The infiltrating growth pattern and likelihood of vascular and perineural involvement distinguishes basal cell adenocarcinoma from basal cell adenoma. Other diagnostic considerations include adenoid cystic carcinoma and basaloid squamous carcinoma. Basal cell adenocarcinomas show strong immunoreactivity to cytokeratin 7 and variable myoepithelial staining with S100. It is necessary to differentiate basal cell adenocarcinoma from other basaloid cell tumors of the minor salivary glands because of the prognosis and potential differences in treatment, particularly adenoid cystic adenocarcinoma and basaloid squamous carcinoma. Surgical excision with a wide margin to ensure complete removal has been suggested as the primary treatment for basal cell adenocarcinoma. Radiotherapy has been proposed for lesions in the minor salivary glands because of the higher likelihood of vascular and neural invasion and for those that are diffusely infiltrative. PMID- 17922608 TI - Clinical skills day: preparing third year medical students for their rural rotation. AB - INTRODUCTION: In order to prepare third year medical students in the Rural Physician Associate Program for a nine-month community-based continuity care experience in rural Minnesota, USA, a clinical skills day that featured human patient simulators and standardized patients was developed. Patients presenting with common urgent and routine primary-care problems were developed and presented using the objective structured clinical examination for teaching. The goals of the day were to: (1) distinguish urgent from non-urgent clinical presentation; (2) use clinical guidelines for making decisions; (3) communicate effectively in stressful situations; and (4) uncover a significant clinical issue with a different presenting complaint. METHODS: Case scenarios were written for a variety of diagnoses in patients with differing ages. Scenarios were both urgent and non-urgent and typical of what might be encountered in primary care. They included: chest pain with bradycardia and pulseless electrical activity; major trauma from an all-terrain vehicle; labor and delivery; acute abdomen (acute appendicitis in a 20 year old and diverticulitis in a 70 year old); anaphylaxis after an influenza vaccination; pediatric upper respiratory infection in which the mother demanded antibiotics; knee injury in a middle-aged man after a weekend of football; heartburn with an underlying significant depression; and X-ray review. The experience occurred in the Interprofessional Education and Resource Center (IERC), where each room was a fully equipped ambulatory examination room with a computer for accessing data and a video camera for central monitoring. Faculty were recruited from the College of Medicine and received an on-line presentation orienting them to the IERC, the teaching model and the scenario assigned to them with supporting evidence-based guidelines. Students reviewed an on-line audio-visual presentation orienting them to the IERC and outlining the learning expectations for the day. Otherwise, students were not expected to prepare for the day because this was an immersion learning experience. Faculty were present in each room as observers, facilitators and educators. Their roles were active or passive, depending on the case scenario and the presence of a simulator or standardized patient. Each station, except the radiology station, involved a debriefing at the end for final questions, and distribution of educational resources or summary teaching points. Standardized patients also gave the students feedback. Students were randomly assigned to small groups of three to four students and rotated through the stations as a unit. RESULTS: To date two classes of students (n = 77) have participated. Evaluations were completed by both students and faculty and included both qualitative and quantitative data immediately after the event and 9 months later (n = 59). Evaluations were overwhelmingly positive with means well above four on a five-point Likert scale. Feedback from both immediate and delayed evaluations were and continue to be used to improve the session for the following year. CONCLUSION: Both students and faculty were enthusiastic about this 'hands on' team learning format, which provided students with opportunities to begin to understand the complex skills that they will need before they learn them step-by-step. PMID- 17922611 TI - Identifying the relative contributions of Rac1 and Rac2 to osteoclastogenesis. AB - Rac small GTPases may play an important regulatory role in osteoclastogenesis. Our in vitro and in vivo results show that both Rac1 and Rac2 are required for optimal osteoclast differentiation, but Rac1 is more critical. Rac1 is the key Rac isoform responsible for regulating ROS generation and the actin cytoskeleton during the multiple stages of osteoclast differentiation. INTRODUCTION: Recent evidence suggests that the Rac small GTPases may play an important regulatory role in osteoclastogenesis. This finding is important because bisphosphonates may regulate their antiresorptive/antiosteoclast effects through the modification of Rho family of small GTPases. MATERIALS AND METHODS: To elucidate the specific roles of the Rac1 and Rac2 isoforms during osteoclastogenesis, we used mice deficient in Rac1, Rac2, or both Rac1 and Rac2 in monocyte/osteoclast precursors. Macrophage-colony stimulating factor (M-CSF)- and RANKL-mediated osteoclastogenesis in vitro was studied by using bone marrow-derived mononucleated preosteoclast precursors (MOPs). The expression of osteoclast specific markers was examined using quantitative real-time PCR and Western blot analysis. Free actin barbed ends in bone marrow MOPs after M-CSF stimulation was determined. The ability of MOPs to migrate toward M-CSF was assayed using Boyden chambers. Margin spreading on heparin sulfate-coated glass and RANKL-induced reactive oxygen species generation were also performed. Functional assays of in vitro-generated osteoclasts were ascertained using dentine sections from narwal tusks. Osteoclast levels in vivo were counted in TRACP and immunohistochemically stained distal tibial sections. In vivo microarchitexture of lumbar vertebrate was examined using microCT 3D imaging and analysis. RESULTS: We show here that, although both Rac isoforms are required for normal osteoclast differentiation, Rac1 deletion results in a more profound reduction in osteoclast formation in vitro because of its regulatory role in pre-osteoclast M-CSF-mediated chemotaxis and actin assembly and RANKL-mediated reactive oxygen species generation. This Rac1 cellular defect also manifests at the tissue level with increased trabecular bone volume and trabeculae number compared with wildtype and Rac2-null mice. This unique mouse model has shown for the first time that Rac1 and Rac2 play different and nonoverlapping roles during osteoclastogenesis and will be useful for identifying the key roles played by these two proteins during the multiple stages of osteoclast differentiation. CONCLUSIONS: Rac1 and Rac2 play different and nonoverlapping roles during osteoclastogenesis. This model showed that Rac1 is the key Rac isoform responsible for regulating ROS generation and the actin cytoskeleton during the multiple stages of osteoclast differentiation. PMID- 17922612 TI - Histomorphometric and microCT analysis of bone biopsies from postmenopausal osteoporotic women treated with strontium ranelate. AB - Strontium ranelate is a new anti-osteoporotic treatment. On bone biopsies collected from humans receiving long-term treatment over 5 yr, it has been shown that strontium ranelate has good bone safety and better results than placebo on 3D microarchitecture. Hence, these effects may explain the decreased fracture rate. INTRODUCTION: Strontium ranelate's mode of action involving dissociation of bone formation and resorption was shown in preclinical studies and could explain its antifracture efficacy in humans. MATERIALS AND METHODS: One hundred forty-one transiliac bone biopsies were obtained from 133 postmenopausal osteoporotic women: 49 biopsies after 1-5 yr of 2 g/d strontium ranelate and 92 biopsies at baseline or after 1-5 yr of placebo. RESULTS AND CONCLUSIONS: Histomorphometry provided a 2D demonstration of the bone safety of strontium ranelate, with significantly higher mineral apposition rate (MAR) in cancellous bone (+9% versus control, p = 0.019) and borderline higher in cortical bone (+10%, p = 0.056). Osteoblast surfaces were significantly higher (+38% versus control, p = 0.047). 3D analysis of 3-yr biopsies with treatment (20 biopsies) and placebo (21 biopsies) using microCT showed significant changes in microarchitecture with, in the strontium ranelate group, higher cortical thickness (+18%, p = 0.008) and trabecular number (+14%, p = 0.05), and lower structure model index (-22%, p = 0.01) and trabecular separation (-16%, p = 0.04), with no change in cortical porosity. The changes in 3D microarchitecture may enhance bone biomechanical competence and explain the decreased fracture rate with strontium ranelate. PMID- 17922610 TI - Lean mass and not fat mass is associated with male proximal femur strength. AB - Obesity is suspected to confer protection against fracture, but evidence is mixed. We examined proximal femur geometry and body composition measures in a diverse group of 1171 men (30-79 yr of age). Analyses showed that nonbone lean mass, but not fat mass, is independently associated with measures of proximal femur density, axial and bending strength, and resistance to buckling. INTRODUCTION: Obesity is often said to confer protection against fracture, but the mechanisms driving such an association remain poorly understood. We hypothesized that the effect of increased body mass on bone structure would be accounted for by total and/or appendicular nonbone lean mass, and that once these trends were removed, fat mass would show no protective influence. To test this hypothesis, we examined body composition and geometric indices of proximal femur strength in an ethnically diverse (black, Hispanic, and white) sample of randomly selected men, 30-79 yr of age. MATERIALS AND METHODS: Data were obtained from N = 1171 community-dwelling subjects enrolled in the cross-sectional Boston Area Community Health/Bone study. Body composition was obtained by DXA. Hip geometry parameters at the narrow neck, intertrochanter, and shaft were obtained using Hip Structural Analysis of DXA images. These measures included BMD, bone material in cross-sections (cross-sectional area), bending strength (section modulus), and propensity to buckle under compression (average buckling ratio). Analyses controlled for age, race/ethnicity, height, and physical activity. RESULTS: In exploratory analyses, lean mass, fat mass, and BMI were each positively associated with hip strength. However, controlling for lean mass was sufficient to remove the positive, and induce a negative, association for fat mass or BMI. Associations between lean mass and hip strength were strongest and resistant to control for other measures. Lean mass alone was sufficient to account for a substantial proportion of racial/ethnic difference in hip strength measures, whereas fat mass exhibited no comparable explanatory power. CONCLUSIONS: The positive association between relative weight and proximal femur strength is accounted for by lean mass, suggesting that, in men, the protective effect of BMI in preventing fracture is mediated not by adipose tissue but by the influence of increased muscle mass accompanying elevated BMI. PMID- 17922613 TI - Clinical and cellular manifestations of OSTM1-related infantile osteopetrosis. AB - Infantile ARO is a genetic disorder characterized by osteoclast dysfunction that leads to osteopetrosis. We describe a novel mutation affecting the OSTM1 locus responsible for ARO. In addition to common clinical features of osteopetrosis, the patient developed a unique neuronal pathology that provided evidence for an essential role of OSTM1 in normal neuronal cell development. INTRODUCTION: Infantile autosomal recessive osteopetrosis (ARO) is a genetic disorder characterized by osteoclast dysfunction that leads to osteopetrosis. We describe a novel mutation affecting the OSTM1 locus responsible for ARO. In addition to common clinical features of osteopetrosis, the patient developed a unique neuronal pathology that provided evidence for an essential role of OSTM1 in normal neuronal cell development. MATERIALS AND METHODS: We report a new case of ARO caused by an homozygous mutation in OSTM1. In addition to osteopetrosis and bone marrow failure, this patient also had neurological impairment not related to bone entrapment. Retinal dystrophy with absent evoked visual potentials and sensorineural deafness were documented, as well as cerebral atrophy and bilateral atrial subependymal heterotopias. RESULTS: The patient developed generalized seizures and had a profound developmental delay. Nerve biopsy failed to show inclusion material suggestive of neuroaxonal dystrophy. Bone marrow transplantation was declined considering the severe neurological compromise. The patient died at 1 yr of age. Osteoclasts derived from peripheral blood were mature and multinucleated. Expression analysis showed that the amount of OSTM1 cDNA transcript was significantly lowered but not absent. CONCLUSIONS: These results support the role of OSTM1 in osteoclast function and activation. However, they also suggest that OSTM1 has a primary role in neural development not related to lysosomal dysfunction. PMID- 17922614 TI - Biological co-adaptation of morphological and composition traits contributes to mechanical functionality and skeletal fragility. AB - A path analysis was conducted to determine whether functional interactions exist among morphological, compositional, and microstructural traits for young adult human tibias. Data provided evidence that bone traits are co-adapted during ontogeny so that the sets of traits together satisfy physiological loading demands. However, certain sets of traits are expected to perform poorly under extreme load conditions. INTRODUCTION: Previous data from inbred mouse strains suggested that biological processes within bone co-adapt morphological and compositional traits during ontogeny to satisfy physiological loading demands. Similar work in young adult humans showed that cortical tissue from slender tibias was stiffer, less ductile, and more susceptible to accumulating damage. Here we tested whether the relationships among morphology and tissue level mechanical properties were the result of biological processes that co-adapt physical traits, similar to those observed for the mouse skeleton. MATERIALS AND METHODS: Cross-sectional morphology, bone slenderness (Tt.Ar/Le), and tissue level mechanical properties were measured from tibias from 14 female (22-46 yr old) and 17 male (17-46 yr old) donors. Physical bone traits measured included tissue density, ash content, water content, porosity, and the area fractions of osteonal, interstitial, and circumferential lamellar tissues. Bivariate relationships among traits were determined using linear regression analysis. A path analysis was conducted to test the hypothesis that Tt.Ar/Le is functionally related to mineralization (ash content) and the proportion of total area occupied by cortical bone. RESULTS: Ash content correlated negatively with several traits including Tt.Ar/Le and marrow area, indicating that slender bones were constructed of tissue with higher mineralization. Path analysis revealed that slender tibias were compensated by higher mineralization and a greater area fraction of bone. CONCLUSIONS: The results suggest that bone adapts by varying the relative amount of cortical bone within the diaphysis and by varying matrix composition. This co-adaptation is expected to lead to a particular set of traits that is sufficiently stiff and strong to support daily loads. However, increases in mineralization result in a more brittle and damageable material that would be expected to perform poorly under extreme load conditions. Therefore, focusing attention on sets of traits and the relationship among traits may advance our understanding of how genetic and environmental factors influence bone fragility. PMID- 17922615 TI - Bone fragility contributes to the risk of fracture in children, even after moderate and severe trauma. AB - We prospectively examined whether the relationship between skeletal fragility and fracture risk in children 9.9 +/- 0.3 (SD) yr is affected by trauma level. Bone size relative to body size and humeral vBMD showed similar inverse relationships with fracture risk, irrespective of whether fractures followed slight or moderate/severe trauma. INTRODUCTION: Fracture risk in childhood is related to underlying skeletal fragility. However, whether this relationship is confined to low-trauma fractures or whether skeletal fragility also contributes to the risk of fracture caused by higher levels of trauma is currently unknown. MATERIALS AND METHODS: Total body DXA scan results obtained at 9.9 yr of age were linked to reported fractures over the following 2 yr in children from the Avon Longitudinal Study of Parents and Children. DXA scan results that were subsequently derived included total body less head (TBLH) bone size relative to body size (calculated from TBLH area adjusted for height and weight) and humeral volumetric BMD (vBMD; derived from subregional analysis at this site). Trauma level was assigned using the Landin classification based on a questionnaire asking about precipitating causes. RESULTS: Of the 6204 children with available data, 549 (8.9%) reported at least one fracture over the follow-up period, and trauma level was assigned in 280 as follows: slight trauma, 56.1%; moderate trauma, 41.0%; severe trauma, 2.9%. Compared with children without fractures, after adjustment for age, sex, socioeconomic status, and ethnicity, children with fractures from both slight and moderate/severe trauma had a reduced bone size relative to body size (1133 cm(2) in nonfractured children versus 1112 cm(2) for slight trauma fractures, p < 0.001; 1112 cm(2) for moderate/severe trauma fractures, p = 0.001) and reduced humeral vBMD (0.494 g/cm(3) in nonfractured children versus 0.484 g/cm(3) for slight trauma fractures, p = 0.036; and 0.482 g/cm(3) for moderate/severe trauma fractures, p = 0.016). CONCLUSIONS: Skeletal fragility contributes to fracture risk in children, not only in fractures caused by slight trauma but also in those that result from moderate or severe trauma. PMID- 17922616 TI - The withdrawal of torcetrapib from drug development: implications for the future of drugs that alter HDL metabolism. AB - In December 2006, Pfizer withdrew torcetrapib, a cholesterol ester transferase protein (CETP) that elevates plasma HDL levels, from further development following an excess in mortality in the active treatment arm of the study. Although torcetrapib successfully elevated HDL levels, significant increases in blood pressure were observed in three surrogate outcome studies that were conducted over the approximate same time period. Two of these studies examined carotid intima-medial thickness and one examined coronary artery atheroma load and none of the studies found a significant benefit in favour of torcetrapib therapy. It is likely that the torcetrapib-induced increase in blood pressure contributed to the apparent adverse effect of the drug on mortality and further studies are needed to determine why this occurred and whether it is a class effect of CETP inhibitors. In addition, further research is needed to determine whether the manner which CETP alters vascular biology and, in particular, the effect that it has on vascular inflammation associated with denuded endothelium. Despite disappointing results so far, CETP inhibitors should not be abandoned as much remains to be learnt from them and they may yet prove to be a valuable class of lipid-modifying drug. PMID- 17922617 TI - Discontinued drugs in 2006: renal, endocrine and metabolic drugs. AB - This perspective summarizes key compounds from the endocrine and metabolic area that were discontinued from clinical development during the calendar year 2006. This is a continuation in a series of perspectives of each of the editorial areas summarized by Expert Opinion on Investigational Drugs. The candidates covered in this summary were being developed for the treatment of diabetes and obesity, as well as for reproductive and urogenital health issues. PMID- 17922618 TI - Next-generation calcineurin inhibitors for ophthalmic indications. AB - Calcineurin inhibitors (CNIs) are potent immunosuppressants that reversibly inhibit T-cell proliferation and prevent the release of pro-inflammatory cytokines by blocking the activity of calcineurin, a ubiquitous enzyme that is found in cell cytoplasm. CNIs can be highly effective in immune-mediated ophthalmic diseases such as uveitis, dry eye syndrome and inflammatory blepharitis, as well as for the prevention of rejection in corneal transplants. ISA-247/LX-211 is a novel CNI that is in Phase III clinical development for the treatment of various forms of non-infectious uveitis. ISA-247/LX-211 is a rationally designed analog of ciclosporin A that exhibits more predictable pharmacokinetic and pharmacodynamic properties and a 4-fold greater calcineurin inhibition than its parent compound, ciclosporin A. ISA-247/LX-211 has been observed to be effective, well-tolerated, and safe in early clinical trials, exhibiting a much wider therapeutic window compared with classic CNIs, such as ciclosporin A and tacrolimus. An alternative approach to widening the therapeutic window for the therapy of ophthalmic disorders lies in local delivery of CNIs through polymeric implants that release the drug over long periods of time. The silicone matrix episcleral implant LX-201 is in Phase III development at present for the prevention of rejection in high-risk cornea transplantation. PMID- 17922619 TI - Novel cardiac myosin activators for acute heart failure. AB - Heart failure remains a leading cause of hospitalisation and death. Treatment of acute heart failure has not improved as rapidly as treatment for chronic heart failure. There are three classes of inotropic agents that are in use at present: the catecholamines, phosphodiesterase inhibitors and calcium sensitisers. Cardiac specific myosin ATPase activators are a novel class of agents designed to improve myocardial contractility by accelerating the productive phosphate-release step of the crossbridge cycle. This article reviews the mechanism of action of myosin ATPase activators, the results of preclinical and Phase I studies and their potential role in the management of heart failure. PMID- 17922620 TI - Future options for imatinib mesilate-resistant tumors. AB - The outcome of patients with gastrointestinal stromal tumors has been dramatically improved by therapy with imatinib mesilate (imatinib mesylate), a KIT and platelet-derived growth factor (PDGFR) tyrosine kinase inhibitor. Unfortunately, the majority of patients eventually experience disease progression due to drug resistance. Recent elucidation of the mechanisms of resistance to imatinib, particularly the acquisition of secondary mutations of the KIT and PDGF receptors, has provided significant insight and potential for the development of novel therapies. This review discusses the efficacy of sunitinib, which is approved for the treatment of patients with imatinib-resistant tumors, and highlights a number of emerging second-generation receptor tyrosine kinase inhibitors that show therapeutic potential in imatinib-resistant patients. Also considered are several promising agents targeting pathways downstream of the constitutionally activated KIT and PDGF receptors. Strategies to overcome imatinib resistance by optimizing combination therapy and selecting specific kinase inhibitors based on the secondary mutations identified in tumors of individual patients are presented. PMID- 17922621 TI - Peroxisome proliferator-activated receptor-gamma ligands as investigational modulators of angiogenesis. AB - PPAR-gamma ligands constitute important insulin sensitizers that have already been approved for the treatment of human metabolic disorders. They also exert pleiotropic effects on cell proliferation and cancer and are now being explored in preclinical studies. Angiogenesis constitutes a multifaceted process that is implicated in tumor development and other benign disease states that are associated with diabetes. Recent data have further extended the crucial role of PPAR-gamma ligands as potential angiogenesis modulators, in vitro and in vivo. This review summarizes the latest knowledge of the role of PPAR-gamma ligands in angiogenesis that are related to both malignant and non-malignant disease states. Taking into careful consideration the data so far, PPAR-gamma could be considered as a therapeutic target for diverse disease states in which excessive angiogenesis is implicated, including cancer and diabetes complications. PMID- 17922622 TI - The therapeutic potential of O6-alkylguanine DNA alkyltransferase inhibitors. AB - Resistance to O(6-)alkylating agents can be overcome by depletion of the DNA repair protein, O(6)-alkylguanine DNA alkyltransferase. Inhibitors of this protein act as pseudosubstrates and, so far, O(6)-benzylguanine and lomeguatrib have been tested in clinical trials. Inherently non-toxic, optimum doses for protein depletion have been established for both agents. Myelosuppression of alkylating agents is significantly enhanced when used in combination with these agents, necessitating significant reductions in standard doses. Consequently, no improvement in efficacy is seen. Strategies to limit myelotoxicity are complex and will be very difficult to apply clinically. O(6)-alkylguanine DNA alkyltransferase inhibition may also potentiate the toxicity of other agents such as cyclophosphamide and irinotecan. Other mechanisms of DNA repair are also important and drugs targeting some of these systems are in early phase clinical trials. PMID- 17922623 TI - Small-molecule inhibitors of PDE-IV and -VII in the treatment of respiratory diseases and chronic inflammation. AB - Targeting phosphodiesterase IV (PDE-IV) with small-molecule inhibitors as a therapeutic for chronic inflammatory disorders has been an active area of research interest for many years. The major drawback, however, has been to develop pharmacophores that would differentiate between targeting isoforms of PDE IV associated with inflammation, as opposed to those that cause emesis, a major side effect associated with PDE-IV inhibition. Several different approaches have been employed, including designing subtype selective PDE-IV inhibitors. A recent approach has been to develop chemotypes that target PDE-VII, a cAMP-specific PDE, expressed widely in immune and pro-inflammatory cells. It is hypothesized that dual inhibitors, which function to inhibit both PDE-IV and VII, may achieve a higher therapeutic index and thereby exhibit a lower propensity to cause adverse side effects that are characteristic when targeting PDE-IV alone. This review focuses on the major classes of compounds that are presently being studied for their potential to inhibit PDE-VII and discusses the available data in the development of dual PDE-IV and -VII inhibitors, their biologic activity and their scope as a therapeutic choice in chronic inflammatory diseases. PMID- 17922624 TI - The anti-inflammatory effect of A3 adenosine receptor agonists: a novel targeted therapy for rheumatoid arthritis. AB - Targeting the A(3) adenosine receptor (A(3)AR) to combat inflammation is a new concept based on two findings. First, A(3)AR is highly expressed in inflammatory cells, whereas low expression is found in normal tissues. This receptor was also found to be overexpressed in peripheral blood mononuclear cells, reflecting receptor status in the remote inflammatory process. Second, A(3)AR activation with a specific agonist induces de-regulation of the NF-kappaB signaling pathway in inflammatory cells, as well as initiation of immunomodulatory effects. The A(3)AR agonist CF-101 (known generically as IB-MECA) induces anti-inflammatory effects in experimental animal models of collagen- and adjuvant-induced arthritis. Combined therapy with CF-101 and methotrexate in adjuvant-induced arthritis rats yielded an additive anti-inflammatory effect. Methotrexate induced upregulation of A(3)AR, rendering the inflammatory cells more susceptible to CF 101. In Phase I and in Phase IIa human studies, CF-101 was safe, well tolerated and showed strong evidence of an anti-inflammatory effect in rheumatoid arthritis patients. In peripheral blood mononuclear cells withdrawn from the patients at base line, a statistically significant correlation between A(3)AR expression level and response to the drug was noted. It is suggested that A(3)AR may serve as a biologic marker to predict patient response to the drug. Taken together, this information suggests that A(3)AR agonists may be a new family of orally bioavailable drugs to be developed as potent inhibitors of autoimmune inflammatory diseases. PMID- 17922625 TI - Oxcarbazepine in neuropathic pain. AB - Neuropathic pain is a frequent condition that can result from a variety of underlying conditions and is frequently chronic and difficult to treat. A number of drugs are used to treat neuropathic pain, including anticonvulsants and antidepressants. Oxcarbazepine, a recently introduced antiepileptic drug, was found to possess antineuralgic properties in animal models of neuropathic pain. Several double-blind, placebo-controlled trials have evaluated oxcarbazepine in painful diabetic neuropathy and trigeminal neuralgia. There is good evidence that oxcarbazepine is effective in relieving the pain associated with trigeminal neuralgia. Its efficacy in treating painful diabetic neuropathy is less clear; however, it seems to be useful when tolerated at doses of 1800 mg/day. PMID- 17922626 TI - Omega-3 fatty acid eicosapentaenoic acid. A new treatment for psychiatric and neurodegenerative diseases: a review of clinical investigations. AB - Decreased n-3 fatty acid levels have been reported in patients with depression, schizophrenia or Alzheimer's disease. Recently, eicosapentaenoic acid (EPA) has been used to treat several psychiatric and neurodegenerative diseases due to its anti-inflammatory and neuroprotective effects. A total of six out of seven clinical trials have shown that EPA significantly improved depressive symptoms when compared with the placebo-treated populations. Several investigations have also reported that EPA could effectively treat schizophrenia. A case report and a clinical trial have shown that EPA was beneficial for the management of most symptoms of Huntington's disease, while a more extensive clinical investigation has demonstrated that EPA could only improve motor functions. Further clinical studies are required to fully explore the effects of EPA on other neurodegenerative diseases. The limitations of previous studies and further research directions have also been discussed. PMID- 17922628 TI - C.E.R.A.: pharmacodynamics, pharmacokinetics and efficacy in patients with chronic kidney disease. AB - C.E.R.A., a continuous erythropoietin receptor activator, has been developed for the treatment of anaemia in patients with chronic kidney disease. Compared with other erythropoiesis-stimulating agents, C.E.R.A. has a unique pharmacological profile, including a longer elimination half-life and slower clearance rate. This allows C.E.R.A. to be administered at extended intervals up to once every month. Phase III clinical trials have shown that C.E.R.A. once every 2 weeks corrects anaemia in erythropoiesis-stimulating agent-naive patients who are on or are not on dialysis, whereas once-monthly C.E.R.A. maintains stable haemoglobin levels when patients are directly converted from more frequent epoetin or darbepoetin alpha administration. C.E.R.A. is well tolerated. This review summarises clinical data on C.E.R.A. and discusses the potential effect of this novel agent on clinical practice. PMID- 17922627 TI - V2 receptor antagonism with tolvaptan in heart failure. AB - The prevalence and incidence of congestive heart failure continues to increase. The two hallmarks of this syndrome, sodium and water retention, are frequently a therapeutic challenge. Most conventional diuretics act primarily as saluretics by inhibiting renal tubular electrolyte reabsorption, which, due to osmotic pressure, promotes excretion of isotonic fluid. The peptide hormone arginine vasopressin vasoconstricts at the V(1A) receptor and promotes water reabsorption via the V(2) receptor in the renal collecting duct by inserting aquaporin-2 water channels into the luminal membrane. Tolvaptan, the first orally available non peptide V(2) receptor antagonist, acts as a potent aquaretic. In this paper, the authors review the pharmacology of tolvaptan and discuss the results of the initial clinical trials with this potent new drug. PMID- 17922629 TI - Bazedoxifene and bazedoxifene combined with conjugated estrogens for the management of postmenopausal osteoporosis. AB - Bazedoxifene acetate (WAY-140424; TSE-424) is an investigational non-steroidal indole-based selective estrogen receptor modulator (SERM) - also classified as an estrogen agonist/antagonist - that is being developed as a daily oral drug for the prevention and treatment of postmenopausal osteoporosis (PMO). Clinical studies have shown favorable effects on the skeleton, with prevention of bone loss in postmenopausal women without osteoporosis and reduction in vertebral fracture risk in women with PMO, without stimulation of endometrium or breast. Bazedoxifene combined with conjugated estrogens is an investigational tissue selective estrogen complex, the first in a new class of therapeutic agents that pairs a selective estrogen receptor modulator with estrogens. Clinical trials with bazedoxifene/conjugated estrogens in postmenopausal women have shown skeletal benefit with improvement in menopausal vasomotor symptoms and little or no stimulation of endometrial or breast tissue. Bazedoxifene/conjugated estrogens is a potential agent for the prevention of PMO and control of menopausal symptoms. PMID- 17922630 TI - Inhaled insulin using AERx insulin Diabetes Management System (AERx iDMS). AB - Diabetes is a chronic, debilitating disease that afflicts millions of people worldwide and poor glycemic control in this disease leads to numerous microvascular and macrovascular complications. There is growing evidence that tight glycemic control prevents the development, and delays the progression, of microvascular complications and possibly macrovascular disease in patients with diabetes. All patients with Type 1 diabetes and many patients with Type 2 diabetes require intensive insulin therapy to achieve optimal glucose control. Although subcutaneous insulin therapy is the mainstay of insulin therapy, there are patients who fear needles and often refuse to start insulin therapy despite suboptimal glucose control. In these patients, inhaled insulin represents a non invasive, painless method to administer intensive insulin treatment. The Novo Nordisk AS AERx iDMS (insulin Diabetes Management System) for inhaled insulin is a novel device that administers an aerosol of liquid insulin into the deep lung with dose adjustments as precise as one subcutaneous unit. Initial pharmacokinetic and pharmacodynamic studies demonstrate that the device delivers liquid insulin in a clear dose-response manner and with a rapid onset of action similar to the fast-acting analog insulins. At present, large, long-term Phase III studies are in progress to document not only the efficacy, but also the safety and feasibility of this device in the treatment of patients with diabetes. PMID- 17922631 TI - Pixantrone: a novel aza-anthracenedione in the treatment of non-Hodgkin's lymphomas. AB - Pixantrone (BBR-2778) is a novel mitoxantrone-like drug, which lacks the 5,8 dihyroxy substitution groups thought to be responsible for the cardiac toxicity associated with mitoxantrone. In Phase I/II single-agent pixantrone clinical trials, neutropenia was the dose-limiting toxicity and the maximum tolerated dose was 150 mg/m(2)/week for 3 weeks every 4 weeks. In relapsed aggressive non Hodgkin's lymphomas, weekly single-agent pixantrone 85 mg/m(2) for 3 weeks every 4 weeks was associated with a 27% overall response and a 15% complete response. When intensively pretreated patients with relapsed aggressive non-Hodgkin's lymphoma were treated with a cyclophosphamide, pixantrone, vincristine and prednisolone regimen (pixantrone substituted for doxorubicin in standard regimen [cyclophosphamide, doxorubicin, vincristine and prednisolone regimen (CHOP)]), the overall response was 74% and complete response 57%. In the BBR-2778, methylprednisolone, cisplatin and cytosine arabinoside (BSHAP) regimen, 58% overall response and 37% complete response were achieved. A number of randomised studies of pixantrone (BBR-2778) in patients with relapsed indolent or aggressive lymphomas are ongoing. PMID- 17922632 TI - The therapeutic role of targeting protein kinase C in solid and hematologic malignancies. AB - The protein kinase C (PKC) family, the most prominent target of tumor-promoting phorbol esters, is functionally linked to cell differentiation, growth, survival, migration and tumorigenesis and so mediates tumor cell proliferation, survival, multidrug resistance, invasion, metastasis and tumor angiogenesis. Therefore, targeting PKC isozymes may represent an attractive target for novel anticancer therapies. Recent preclinical and clinical studies using the macrocyclic bisindolylmaleimide enzastaurin or the N-benzylstaurosporine midostaurin demonstrate promising activity of PKC inhibitors in a variety of tumors, including diffuse large B-cell lymphoma, multiple myeloma and Waldenstroem's macroglobulinemia. However, our knowledge of PKCs in tumorigenesis is still only partial and each PKC isoform may contribute to tumorigenesis in a distinct way. Specifically, PKC isoforms have vastly different roles, which vary depending on expression levels of organ and tissue distribution, cell type, intracellular localization, protein-protein and lipid-protein interactions and the biologic environment. Although PKC activation generally positively affects tumor cell growth, motility, invasion and metastasis, recent reports show that many PKCs can also have negative effects. Therefore, it is necessary to further dissect the relative contribution of PKC isozymes in the development and progression of specific tumors in order to identify therapeutic opportunities, using either PKC inhibitors or PKC activators. PMID- 17922633 TI - Oral beclomethasone dipropionate: a topically active corticosteroid for the treatment of gastrointestinal graft-versus-host disease following allogeneic hematopoietic cell transplantation. AB - Beclomethasone dipropionate (BDP) is a topically active anti-inflammatory corticosteroid. Oral BDP is metabolized in the intestine to a potent metabolite, 17-beclomethasone monopropionate (17-BMP). An oral formulation (orBec; DOR BioPharma), consisting of a gastric release and an enteric-coated pill, was studied in patients with acute gastrointestinal graft-versus-host disease, an inflammatory disorder that is common after allogeneic hematopoietic cell transplantation. Randomized trials demonstrated that orBec is safe and effective in treating acute gastrointestinal graft-versus-host disease (GVHD) when used with a short induction course of prednisone, reducing the risk of GVHD treatment failure by > 60% and reducing mortality 1 year after randomization by 45%, with fewer deaths due to infection and recurrent malignancy. The type of conditioning and the type of donor had no effect on the frequency of GVHD treatment failure during the 80-day study period; the greatest benefit in terms of survival was among patients who had received reduced-intensity conditioning therapy and among those who received a graft from other than a human leukocyte antigen-matched sibling. orBec controls the intestinal inflammatory process of GVHD and avoids prolonged exposure to prednisone, which is the present standard of care. Oral BDP is the only therapy to be studied in the last 30 years to effectively treat acute GVHD and reduce mortality. PMID- 17922634 TI - 14th Annual Meeting of the Psychoneuroimmunology Research Society 28 - 29 May 2007, Bordeaux, France. AB - The Association for Neurophyschopharmacology hosted a satellite meeting as part of the 14th Annual Meeting of the Psychoneuroimmunology Research Society. The meeting was entitled 'Cytokines and Depression III: Identification and Treatment of Symptoms Associated with Inflammation in Diseases with Inflammation in Medically Ill Patients' and was held in Bordeaux, France on 28 - 29 May, 2007. The meeting comprised approximately 40 participants from many leading laboratories and hospitals from around the world looking to understand some of the clinical issues associated with depression and behavioural changes, with the aims of exploring better ways of clinical monitoring and marshalling drug discovery efforts from bespoke and alternate indications in providing new therapeutic approaches. PMID- 17922635 TI - 67 th annual meeting of the American Diabetes Association. AB - The 67 th meeting of the American Diabetes Association was held in Chicago on 22 26 June. This annual meeting continues to grow in size and scope and is a unique combination of basic science and medical science but also incorporates all aspects of healthcare and pharmaceutical business relating to the treatment of diabetes. The meeting was composed of general sessions, symposia summarizing the status of various fields of study and medical practice, together with both oral and poster presentations of new, previously unpublished research. The abstracts are published in Diabetes and a collection of the information can be found online with very useful summaries from the final day. These contain personalized summaries of key findings of the meetings as seen by key researches in the field. In this Meeting Highlights article, the key take-away messages are summarized from the author's point of view. PMID- 17922636 TI - Therapeutic management of ocular adnexal MALT lymphoma. AB - Non-Hodgkin lymphomas constitute 50% of all orbital malignancies. Mucosa associated lymphoid tissue (MALT)-type lymphoma is the most common histology category in this anatomic region. Ocular adnexal lymphoma of MALT-type is an indolent and rarely lethal malignancy that can often be managed with observation alone. Occasionally, lymphomatous lesions determine the symptoms that condition a patient's quality of life, and so require an immediate antineoplastic treatment. Several strategies are available, but reliable indications supported by prospective trials do not exist. Some therapeutic strategies are associated with substantial side effects and require a well-balanced therapeutic decision, which should take into account several variables related to the patient, the lymphoma, and the treatment itself. In this review, the authors analyze related literature and propose therapeutic guidelines for the management of ocular adnexal lymphoma of MALT-type. PMID- 17922637 TI - Lipid second messengers and cell signaling in vascular wall. AB - Agonists of cellular receptors, such as receptor tyrosine kinases, G protein coupled receptors, cytokine receptors, etc., activate phospholipases (C(gamma), C(beta), A(2), D), sphingomyelinase, and phosphatidylinositol-3-kinase. This produces active lipid metabolites, some of which are second messengers: inositol trisphosphate, diacylglycerides, ceramide, and phosphatidylinositol 3,4,5 trisphosphate. These universal mechanisms are involved in signal transduction to maintain blood vessel functions: regulation of vasodilation and vasoconstriction, mechanical stress resistance, and anticoagulant properties of the vessel lumen surface. Different signaling pathways realized through lipid second messengers interact to one another and modulate intracellular events. In early stages of atherogenesis, namely, accumulation of low density lipoproteins in the vascular wall, cascades of pro-atherogenic signal transduction are triggered through lipid second messengers. This leads to atherosclerosis, the general immuno-inflammatory disease of the vascular system. PMID- 17922638 TI - Free radical oxidation of proteins and its relationship with functional state of organisms. AB - Most reactive oxygen species (ROS) in living organisms are produced as byproducts of many processes. Being highly active, ROS interact with virtually all cellular components particularly modifying their properties. In this review, detailed analysis of chemical modifications of proteins on their interaction with ROS is given with particular interest in cleavage of polypeptide chains and oxidation of side chains of amino acid residues. Special attention has been paid to identification of products of free radical modification of proteins with a focus on the formation of additional carbonyl groups, which are the most frequently used markers of these processes. Functional consequences of protein modification by ROS depend on the nature of ROS and protein as well as particular conditions of their interaction. The relationship between protein oxidation and functional state of organisms, particularly aging, hyperoxia and hypoxia, and heat shock, as well as with different pathologies has been analyzed. The final part of the article is devoted to possible ways of protecting proteins against oxidation in vivo. PMID- 17922639 TI - Kinetic models of cyclooxygenase and peroxidase inactivation of prostaglandin-H synthase during catalysis. AB - Kinetic models of inactivation of cyclooxygenase and peroxidase activities of prostaglandin-H-synthase (PGHS) during cyclooxygenase and peroxidase reactions catalyzed by the enzyme and also on preincubation with H2O2 have been developed; these models account for data obtained by the authors as well as data from the literature. Being rather simple, these models simultaneously describe the processes of cyclooxygenase and peroxidase inactivation of PGHS, using the minimal set of experimental parameters. PMID- 17922640 TI - Investigation of chimerical and tagged forms of recombinant rat nucleoside diphosphate kinases alpha and beta. Interaction with rhodopsin-transducin complex and thermal stability. AB - To elucidate the physicochemical basis of differences between the isoforms of mammalian multifunctional nucleoside diphosphate kinase (NDP), we investigated the recombinant rat homohexameric NDP kinases alpha and beta, consisting of highly homologous alpha or beta subunits of 152 residues each and differing only in variable regions V1 and V2, and their chimerical forms (NDP kinase alpha(1 130)beta(131-152) and NDP kinase beta(1-130)alpha(131-152)) and tagged derivatives (NDP kinase HA-alpha(1-130)beta(131-152), NDP kinase HA-beta(1 130)alpha(131-152), and NDP kinase HA-beta). The thermal stability of these proteins and the ability of some of them to interact with the rhodopsin transducin (R*Gt) complex have been studied. It was found that NDP kinase alpha, NDP kinase alpha(1-130)beta(131-152), and NDP kinase HA-alpha(1-130)beta(131-152) were similar in their thermal stability (T(1/2) = 61-63 degrees C). NDP kinase beta, NDP kinase beta(1-130)alpha(131-152), NDP kinase HA-beta(1-130)alpha(131 152), and NDP kinase HA-beta were inactivated at a lower temperature (T(1/2) = 51 54 degrees C). NDP kinase HA-alpha(1-130)beta(131-152) interacted with the R*Gt complex in the same manner as NDP kinase alpha, whereas the interaction of NDP kinase HA-beta(1-130)alpha(131-152) and NDP kinase beta with the photoreceptor membranes under the same conditions was very weak. It is suggested that the variability of the region V1 is a structural basis for the multifunctionality of NDP kinase hexamers in the cell. PMID- 17922642 TI - Regulation of matrix metalloproteinase-9 transcription in squamous cell carcinoma of uterine cervix: the role of human papillomavirus gene E2 expression and activation of transcription factor NF-kappaB. AB - Matrix metalloproteinase-9 (MMP-9) plays an important role in initiation and progression of squamous cell carcinoma (SCC) of human uterine cervix. Regulation of MMP-9 expression in such tumors is insufficiently studied. Involvement of the human papillomavirus (HPV) gene E2 and transcription factor NF-kappaB in the regulation of MMP-9 transcription has been shown in some model systems and types of malignant tumors. The present work was mainly designed to reveal a possible role of the HPV gene E2 and transcription factor NF-kappaB in the induction of MMP-9 expression in SCC. Specimens of tumor and corresponding adjacent normal tissue from 26 patients with SCC of the uterine cervix were studied. The intact E2 frame was observed in 19 of 26 (73.1%), the E2 gene mRNA was expressed in 10 of 15 (66.7%), NF-kappaB was activated in 17 of 23 (73.9%), and the expression of MMP-9 mRNA was recorded in 10 of 20 (50%) of the informative cases. The MMP-9 transcription did not correlate with gene E2 status, but in all cases correlated with the activation of NF-kappaB transcription factor (10 of 10 vs. 5 of 10 MMP-9 negative cases, p = 0.016). Thus, the NF-kappaB role has been proved in the regulation of MMP-9 transcription in SCC. There was no correlation of the E2 status and MMP-9 expression with clinical/morphological characteristics of the tumors: size, local invasiveness, metastasizing into regional lymph nodes, and level of differentiation. The high intensity of NF-kappaB activation correlated with low degree of differentiation of the tumors studied (p = 0.044). These findings suggested that NF-kappaB should be a molecular factor of the poor prognosis of human SCC. PMID- 17922641 TI - Tim18, a component of the mitochondrial translocator, mediates yeast cell death induced by arsenic. AB - Evidence is presented that Tim18, a mitochondria translocase, plays a role in the previously described apoptosis induced by arsenite in Saccharomyces cerevisiae. Tim18 deletion mutant exhibited resistance to arsenite. After arsenite treatment, both the wild type and Tim18-deficient cells showed reactive oxygen species (ROS) production. Arsenite induced the higher expression of tim18 in wild type yeast cells. We found that the tim18 deletion mutant also exhibited resistance to other apoptotic stresses such as acetic acid, H2O2, and hyperosmotic stress. These results suggest that Tim18 is important for yeast cell death induced by arsenic, and it may act downstream of ROS production. PMID- 17922643 TI - Pentaspan membrane glycoprotein, prominin-1, is involved in glucose metabolism and cytoskeleton alteration. AB - This study constitutes the first report revealing the participation of prominin-1 in glucose metabolism and cytoskeleton alteration. Upon stimulation with high glucose, the expression of prominin-1 was up-regulated with concomitant down regulation of its phosphorylation. Prominin-1 mutated at its phosphorylation site inflicted a global change in the expression of several genes associated with glucose metabolism in L6 myotube cells. Further, the over-expression of prominin 1 promoted glucose uptake in these cells. Prominin-1 undergoes sustained repression of its expression during confluence and differentiation of L6 myotube cells. The expression of prominin-1 in the MDCK cell modulated cell morphology and promoted cellular migration. These data imply that prominin-1 is involved in glucose metabolism and may regulate cellular glucose through its effect on cytoskeleton. PMID- 17922644 TI - A protein whose binding to Na,K-ATPase is regulated by ouabain. AB - Immunoprecipitation of Na,K-ATPase from kidney homogenate by antibodies against alpha1-subunit results in the precipitation of several proteins together with the Na,K-ATPase. A protein with molecular mass of about 67 kD interacting with antibodies against melittin (melittin-like protein, MLP) was found in the precipitate when immunoprecipitation was done in the presence of ouabain. If immunoprecipitation was done using antibodies against melittin, MLP and Na,K ATPase alpha1-subunit were detected in the precipitate, and the amount of alpha1 subunit in the precipitate was increased after the addition of ouabain to the immunoprecipitation medium. MLP was purified from mouse kidney homogenate using immunoaffinity chromatography with antibodies against melittin. The addition of MLP to purified FITC-labeled Na,K-ATPase decreases fluorescence in medium with K+ and increases it in medium with Na+. The enhancement of fluorescence depends upon the MLP concentration. The N-terminal sequence of MLP determined by the Edman method is the following: HPPKRVRSRLNG. No proteins with such N-terminal sequence were found in the protein sequence databases. However, we revealed five amino acid sequences that contain this peptide in the middle part of the chain at distance 553 amino acids from the C-terminus (that corresponds to protein with molecular mass of about 67 kD). Analysis of amino acid sequence located between C terminus and HPPKRVRSRLNG in all found sequences has shown that they were highly conservative and include WD40 repeats. It is suggested that the 67-kD MLP either belongs to the found protein family or was a product of proteolysis of one of them. PMID- 17922645 TI - Identification of leukocyte cationic proteins that interact with ceruloplasmin. AB - Proteins from leukocytes were investigated for their ability to interact with ceruloplasmin (Cp), a copper-containing glycoprotein of human plasma. Extract from leukocytes was subjected to affinity chromatography on Cp-Sepharose, after which proteins were eluted from the resin with 0.5 M NaCl in Tris-HCl, pH 7.4. SDS-PAGE of the eluate revealed protein bands with molecular weights 78, 57, 40, 30, 16, and 12 kD. Among these, Western blotting detected myeloperoxidase (57, 40, and 12 kD) and lactoferrin (78 kD). Also, the 30-kD component had a sequence (1)I-(2)I/V-(3)G-(4)G-(5)R/H at the N-terminus that is likely to indicate the presence of neutrophilic elastase, cathepsin G, proteinase 3, and azurocidin (CAP 37) - all from the family of serprocidins. Mass spectrometry of tryptic fragments indicated the presence of the 16-kD eosinophilic cationic protein (seven peptides), 27-kD cathepsin G (eleven peptides), 27-kD azurocidin (eight peptides), 29-kD neutrophilic elastase (seven peptides), and 27-kD proteinase 3 (six peptides). Myeloperoxidase was represented by 57-, 40-, and 12-kD fragments (thirteen, ten, and four peptides, respectively). Thus, interaction with Cp of five cationic proteins, i.e. of eosinophilic cationic protein, cathepsin G, neutrophilic elastase, proteinase 3, and azurocidin is reported for the first time. PMID- 17922647 TI - Fluorescent DNA probes: study of mechanisms of changes in spectral properties and features of practical application. AB - Spectral data and nucleic acid complex formation properties for more than 30 both newly synthesized and widely used fluorescent nucleotide-specific compounds of various classes have been analyzed. These include phenylbenzene, bisbenzimidazole, psoralen, angelicin, tetrahydrocarbazole, oxophenoxazine, and others. The main rules of a generalized model adequately explaining changes in fluorescent properties of synthetic, low molecular weight nucleotide-specific dyes depending on their chemical structure, mode of interaction with substrate, properties of assay medium, etc. are proposed. Fluorescent nucleotide-specific dyes have been originally used in newly developed methods for: express evaluation of "generalized microbial dissemination" of liquid media; evaluation of possible genotoxic effects of various foodstuffs, pharmaceutical drugs, hazardous environmental factors (including their combined effects on living organisms), etc. PMID- 17922646 TI - Study of interaction of XRCC1 with DNA and proteins of base excision repair by photoaffinity labeling technique. AB - The X-ray repair cross-complementing group 1 (XRCC1) protein plays a central role in base excision repair (BER) interacting with and modulating activity of key BER proteins. To estimate the influence of XRCC1 on interactions of BER proteins poly(ADP-ribose) polymerase 1 (PARP1), apurinic/apyrimidinic endonuclease 1 (APE1), flap endonuclease 1 (FEN1), and DNA polymerase beta (Pol beta) with DNA intermediates, photoaffinity labeling using different photoreactive DNA was carried out in the presence or absence of XRCC1. XRCC1 competes with APE1, FEN1, and PARP1 for DNA binding, while Pol beta increases the efficiency of XRCC1 modification. To study the interactions of XRCC1 with DNA and proteins at the initial stages of BER, DNA duplexes containing a photoreactive group in the template strand opposite the damage were designed. DNA duplexes with 8-oxoguanine or dihydrothymine opposite the photoreactive group were recognized and cleaved by specific DNA glycosylases (OGG1 or NTH1, correspondingly), although the rate of oxidized base excision in the photoreactive structures was lower than in normal substrates. XRCC1 does not display any specificity in recognition of DNA duplexes with damaged bases compared to regular DNA. A photoreactive group opposite a synthetic apurinic/apyrimidinic (AP) site (3-hydroxy-2 hydroxymethyltetrahydrofuran) weakly influences the incision efficiency of AP site analog by APE1. In the absence of magnesium ions, i.e. when incision of AP sites cannot occur, APE1 and XRCC1 compete for DNA binding when present together. However, in the presence of magnesium ions the level of XRCC1 modification increased upon APE1 addition, since APE1 creates nicked DNA duplex, which interacts with XRCC1 more efficiently. PMID- 17922648 TI - Spectral study of the interaction of DNA with benzothiazolyl-benz-alpha-chromene. AB - Absorption and luminescence excitation and emission spectra of newly synthesized 2-(4-methylphenylimino)-3-(2 -benzothiazolyl)benz-alpha-chromene (BCBT) have been studied in the presence of various DNA concentrations. BCBT is characterized by the existence of two different fluorescent systems, exhibiting radiationless fluorescence resonance energy transfer between them. In the range of molar ratios of polynucleotide/dye concentrations from 0 to 50, BCBT preferentially intercalates into DNA due to its benz-alpha-chromene fragment, whereas the 2 benzothiazolyl fragment is responsible for fluorescence. PMID- 17922649 TI - DNA mimicry by proteins as effective mechanism for regulation of activity of DNA dependent enzymes. AB - Modern concepts on mechanisms of DNA-dependent enzyme regulation involving specific DNA-mimicking proteins are considered. There are proteins that share structural resemblance with DNA duplexes. These include inhibitors of type I restriction-modification enzymes (Ocr and ArdA), inhibitors of DNA gyrase MfpA and QnrABS, etc. We describe here structural features of these proteins and mechanisms responsible for their interaction with DNA-dependent enzymes and then discuss perspectives of use of DNA-mimicking proteins in analysis of replication, repair, recombination, mechanisms underlying resistance to antibiotics, and also fields of applied biotechnology. PMID- 17922651 TI - Steady-state kinetics of bifunctional enzymes. Taking into account kinetic hierarchy of fast and slow catalytic cycles in a generalized model. AB - A steady-state approximation of the generalized two-dimensional model of a bifunctional enzyme catalyzing independent proceeding of two one-pathway reactions is considered in a case of mutual influence of the active sites. Coexistence of fast and slow catalytic cycles in the reaction mechanism is analyzed. Conditions when the hierarchy of fast and slow catalytic cycles allows simplification of a two-dimensional model and its reduction to the one dimensional cyclic schemes were determined. Kinetic equations describing these simplified schemes are presented. PMID- 17922650 TI - Cell adhesion proteins and alpha-fetoprotein. Similar structural motifs as prerequisites for common functions. AB - This review summarizes and analyzes data on structural and functional relationships between cell adhesion proteins and alpha-fetoprotein (AFP), which play an important role in embryo- and carcinogenesis and act in synergism with growth factors. These two groups of proteins are mosaic, multimodular, and polyfunctional, and each of their modules can function independently through binding with its specific membrane receptor. Most cell adhesion proteins contain modules similar to epidermal growth factor (EGF) and also their repeats, which determine the involvement of these proteins in regulation of cell proliferation, differentiation, and apoptosis. These EGF-like modules are found to include short motifs similar to the fragment LDSYQCT of human AFP. Both direct and inverted AFP like motifs are linked through a consensus octapeptide motif CXXGY/FXGX. Such AFP like motifs of cell adhesion proteins and the tripeptide RGD found in AFP may be structural prerequisites for common functions of these groups of nonhomologous and unrelated proteins. PMID- 17922652 TI - Molecular oxygen (a substrate of the cyclooxygenase reaction) in the kinetic mechanism of the bifunctional enzyme prostaglandin-H-synthase. AB - Prostaglandin-H-synthase is a bifunctional enzyme catalyzing conversion of arachidonic acid into prostaglandin H2 as a result of cyclooxygenase and peroxidase reactions. The dependence of the rate of the cyclooxygenase reaction on oxygen concentration in the absence and in the presence of electron donor was determined. A two-dimensional kinetic scheme accounting for independent proceeding and mutual influence of the cyclooxygenase and peroxidase reactions and also for hierarchy of the rates of these reactions was used as a model. In the context of this model, it was shown that there are irreversible stages in the mechanism of the cyclooxygenase reaction between points of substrate donation (between donation of arachidonic acid and the first oxygen molecule and also between donation of two oxygen molecules). PMID- 17922653 TI - Transcription factors NFAT2 and Egr1 cooperatively regulate the maturation of T lymphoma in vitro. AB - We have demonstrated that transcription factors Egr1 and NFAT2 cooperate in regulation of the early stages of T-lymphocyte development, whereas the related factors Egr2 and Egr3 do not cooperate with NFAT2. Egr1 and NFAT2 are shown to cooperatively control gene expression of the regulatory factor Id3 and recombinase Rag2, whose functions are critical for T-lymphocyte differentiation. Thus, the concerted action of the transcription factors Egr1 and NFAT2 can play a crucial role in regulation of the T cell differentiation in vitro due to the cooperative regulation of Id3 and Rag2 gene expression. PMID- 17922654 TI - Quenching of the fluorescence of Tyr and Trp residues of firefly luciferase from Luciola mingrelica by the substrates. AB - Luciferase of the firefly Luciola mingrelica is characterized by fluorescence of not only the unique Trp residue (lambda(em) = 340 nm), but also that of Tyr residues (lambda(em) = 308 nm). Quenching of the intrinsic fluorescence of the luciferase by its substrates luciferin and ATP (AMP) has been studied. Luciferin (LH2) quenches Trp fluorescence more efficiently than the fluorescence of Tyr residues. Two centers of quenching of Tyr fluorescence by ATP have been found corresponding apparently to the allosteric and active sites of the luciferase with K(s(ATP)) = 20 and 110 microM, respectively. The influence of one substrate on the affinity of luciferase to the second was investigated using fluorescence. ATP (AMP) binding to the allosteric sites of the luciferase significantly affects the affinity of luciferase to LH2. Formation of the complex between the luciferase and LH2 affects the affinity of both allosteric and active sites of the luciferase to ATP (AMP). The observed effects are probably connected with conformational changes in the luciferase molecule upon its interaction with the substrates. PMID- 17922655 TI - Measuring the pH dependence of hydrogenase activities. AB - The pH dependences of activities of homogenous hydrogenases of Thiocapsa roseopersicina and Desulfomicrobium baculatum in the reaction of hydrogen uptake in solution in the presence of benzyl viologen and the pH dependences of catalytic currents of hydrogen oxidation by electrodes on which these hydrogenases were immobilized were compared. Maximal activities of the hydrogenases from T. roseopersicina and D. baculatum in the reaction hydrogen uptake in solution were observed at pH 9.5 and 8.5, respectively. However, the steady-state current caused by catalytic uptake of hydrogen was maximal for the T. roseopersicina hydrogenase-containing electrode at pH 5.5-6.5 under overvoltage of 30-60 mV, whereas for electrodes with D. baculatum hydrogenase it was maximal at pH 6.0-6.5. Analysis of these data suggests that pH-dependent changes in the hydrogenase activities in solution during hydrogen uptake are due not only to the effect of proton concentration on the enzyme conformation or protonation of certain groups of the enzyme active center, but they are rather indicative of changes in free energy of the reaction accompanying changes in pH. PMID- 17922656 TI - Differential effect of growth factors on hyaluronan synthase gene expression in fibroblasts exposed to oxidative stress. AB - The aim of this study was to evaluate how growth factors (PDGF-BB, EGF, and TGF 1beta) modulate hyaluronan synthase (HAS) activities in normal or stressed cultured human skin fibroblasts. The effects of concomitant treatment with cytokines and FeSO4 plus ascorbate on HAS mRNA expression, protein synthesis, and hyaluronic acid (HA) concentrations were also studied. Treatment of fibroblasts with growth factors up-regulated HAS gene expression and increased HAS enzymes and HA production. PDGF-BB induced HAS mRNA expression, protein synthesis, and HA production more efficiently than EGF and TGF-1beta. EGF was less effective than TGF-1beta. In addition, TGF-1beta reduced the expression and synthesis of HAS3, while PDGF-BB and EGF had the opposite effect. Concomitant treatment with growth factors and the oxidant was able to further increase HAS mRNA expression, once again with the exception of HAS3 with TGF-1beta. HAS protein synthesis was reduced, while HA levels were unaffected in comparison to those obtained from exposure to FeSO4 plus ascorbate alone. In conclusion, although growth factors plus the oxidant synergistically induced HAS mRNA expression in part, enzyme production was not correlated with this increase. Moreover, the increase in HAS mRNA levels was not translated into a consequent rise in HA concentration. PMID- 17922658 TI - Ionogenic groups in the active site of lysostaphin. Kinetic and thermodynamic data compared with x-ray crystallographic data. AB - The active site of lysostaphin is shown to contain a residue of glutamic acid. As judged by a pK value of 9.2 (with pentaglycine bridges in peptidoglycan of staphylococci as a substrate), another ionogenic residue could be the epsilon amino group of a lysine. However, the pH value near a negatively charged cell is supposed to be strongly shifted to acidity as compared to the pH of the solution volume. This shifts the enzyme pH dependence curve in solution to alkalinity. Therefore, the other group might be histidine, which is consistent with the X-ray crystallographic data. A similar shift is likely to occur for lysozyme in the case of Micrococcus lysodeikticus cells. Determination of pK of ionogenic groups in the active sites of alkaline enzymes responsible for lysis of negatively charged bacterial cells gives their apparent values because the "pericellular" and "voluminous" values of pH are not coincident. PMID- 17922657 TI - Extrathymic rearrangement of alphabetaT-lymphocyte antigen receptor genes during pregnancy. AB - The existence of alphabetaT-lymphocyte differentiation processes have been demonstrated in mouse peripheral lymphoid organs during pregnancy. Study of pregnant Swiss mice has shown that the development of the second half of gestation is accompanied by expression of RAG-1 recombinase mRNA and unrearranged TCR alpha-chain (pre-TCRalpha) preferentially in T-lymphocytes of lymph nodes involved in uterine drainage (para-aortal lymph nodes), and to a lesser extent in other lymph nodes (mainly from axillary lymph nodes). The data suggest that during pregnancy the differentiation of alphabetaT lymphocytes may occur not only in central (thymus) but also in peripheral lymphoid organs. PMID- 17922659 TI - Dynamic spin label study of the barstar-barnase complex. AB - The dynamic spin label method was used to study protein-protein interactions in the model complex of the enzyme barnase (Bn) with its inhibitor barstar. The C40A mutant of barstar (Bs) containing a single cysteine residue was modified with two different spin labels varying in length and structure of a flexible linker. Each spin label was selectively bound to the Cys82 residue, located near the Bn-Bs contact site. The formation of the stable protein complex between Bn and spin labeled Bs was accompanied by a substantial restriction of spin label mobility, indicated by remarkable changes in the registered EPR spectra. Order parameter, S, as an estimate of rapid reorientation of spin label relative to protein molecule, was sharply increasing approaching 1. However, the rotational correlation time tau for spin-labeled Bs and its complex with Bn in solution corresponded precisely to their molecular weights. These data indicate that both Bs and its complex with Bn are rigid protein entities. Spin labels attached to Bs in close proximity to an interface of interaction with Bn, regardless of its structure, undergo significant restriction of mobility by the environment of the contact site of the two proteins. The results show that this approach can be used to investigate fusion proteins containing Bn or Bs. PMID- 17922660 TI - Interaction between ACC synthase 1 and 14-3-3 proteins in rice: a new insight. AB - In this study, the interaction between rice 14-3-3 protein and 1 aminocyclopropane carboxylic acid synthase (ACS) was observed in yeast cells using yeast two-hybrid assays. Given the fact that 14-3-3 proteins generally bind to their target proteins in a phosphorylation-dependent manner, a hypothesis regarding the regulatory role of 14-3-3 proteins in the activation of ACS is proposed in which 14-3-3 proteins may bind to the phosphorylated C-terminal tails of ACSs and help them to escape from their fated degradation when ethylene biosynthesis is needed. It is reasonable to believe that 14-3-3 protein may play an important role in regulating ACS activity. PMID- 17922661 TI - Modification of polysaccharides from callus culture of Silene vulgaris (M.) G. using carbohydrases in vitro. AB - Polysaccharides (pectin and intracellular and extracellular arabinogalactans) were isolated from campion callus culture cultivated on medium with varied concentrations of pectinase and beta-galactosidase. A decrease in contents of arabinose residues in pectin and arabinogalactans and of galactose residues in arabinogalactans was associated with an increase in the activities of alpha-L arabinofuranosidase and beta-galactosidase upon addition of pectinase into the medium. Pectinase destroyed the high-molecular-weight (more than 300 kD) fraction of pectin and decreased the content of galacturonic acid residues. alpha-L Arabinofuranosidase transformed arabinogalactan into galactan, and galactan was destroyed under the influence of galactosidase. The contents of arabinogalactan and/or galactan in the cells were decreased, and it was released into the culture medium. Pectin samples with low contents of arabinose and galactose in the side chains and galactan samples were obtained from the callus grown on the medium with beta-galactosidase. Cultivation of the plant cells on medium containing carbohydrases resulted in modification of pectin and arabinogalactan of the cell walls. PMID- 17922662 TI - Expression of different proteoglycans in human breast tumors. AB - The composition of proteoglycans and their changes during malignant transformation are important factors influencing adhesive properties and mitotic activity of tumor cells. In this study, expression level of different proteoglycans (decorin, syndecan-1, lumican, glypican-1, and aggrecan) in tumors and normal human breast tissue was investigated. Multiplex RT-PCR data revealed different expression changes for different proteoglycans in human breast tumors- syndecan expression was activated compared to almost no expression in normal breast tissue, expression of decorin and lumican decreased 2-5- and 2-3-fold, respectively, and aggrecan transcription seems to be unaffected. A change of expression level of decorin correlated with expression of D-glucuronyl-C5 epimerase, a key enzyme responsible for the biosynthesis of idurone-containing glycosaminoglycans, possessing antimitotic activity. The results suggest that changes in decorin, lumican, and syndecan-1 expression in tumor tissue could induce a distortion of proteoglycan composition and mitotic activity of cells in human breast tumor. PMID- 17922663 TI - A new type of subchloroplast fragments isolated from pea chloroplasts in the presence of digitonin. AB - Heavy fragments were isolated from pea chloroplasts using digitonin treatment and differential centrifugation. The particles were characterized by a significantly lowered chlorophyll a/b ratio, contents of photosystem I (PS I) proteins and ATPase, as well as of amount of P700. The content of photosystem II (PS II) proteins decreased insignificantly, whereas that of proteins of the light harvesting complex II did not change. The absorption and low-temperature fluorescence spectra were indicative of a decreased content of PS I. Electron microscopy of ultrathin sections of heavy fragment preparations identified them as grana with reduced content of thylakoids. The diameter of these particles was practically the same as within chloroplasts. Comparison of various characteristics of the fragments and chloroplasts from which the fragments were isolated allowed us to define a high degree of preservation of marginal regions in thylakoids present in the heavy fragment particles. Analysis of the results shows that the procedure of fragmentation produces grana with high extent of thylakoid integrity. The phenomenon of reduction of the thylakoid content in grana, occurring as our heavy fragments, is considered in the frame of our previous hypothesis concerning the peculiarities of grana organization in the transversal direction. PMID- 17922664 TI - Understanding the mechanisms of action of methotrexate: implications for the treatment of rheumatoid arthritis. AB - Methotrexate has been widely used for the treatment of rheumatoid arthritis (RA). The mechanisms of action of methotrexate are complex. Developed as a folic acid analogue, methotrexate inhibits purine and pyrimidine synthesis, which accounts for its efficacy in the therapy of cancer as well as for some of its toxicities. Recently, many studies have focused on the adenosine-mediated antiinflammatory effects of methotrexate. Certain aspects of methotrexate toxicities are also attributed to adenosine release. A better understanding of the mechanisms of action and toxicities of methotrexate will direct clinicians in their treatment approach and toxicity monitoring. Toward that objective, the latest developments in the pharmacokinetics, mechanism of action, pharmacogenetics, and toxicity of methotrexate are herein discussed. PMID- 17922665 TI - Personalized medicine in rheumatoid arthritis: hopes and challenges. AB - Clinicians have an increasing number of therapeutic agents available for the treatment of rheumatoid arthritis (RA). Pharmacogenetics, the study of genetic variation underlying differential response to drugs, seeks to improve treatment of individual patients. Multiple markers of treatment response have been analyzed in RA, but many of the studies are small and retrospective in nature. There are many obstacles to personalized medicine for RA patients, including incomplete understanding of disease pathogenesis, effecting changes in physician behavior, and acceptance of costs by health insurers. Despite these many obstacles, there are many reasons for optimism for future personalized medicine in RA. There have been remarkable advances in genomics, proteomics, and statistical analyses of large amounts of data. The goal of identifying genetic, serum, and clinical factors that allow profiling of individual patients to predict optimal treatment regimens is a worthy pursuit which will hopefully improve clinical care of RA patients. PMID- 17922666 TI - TNF antagonist safety in rheumatoid arthritis: updated evidence from observational registries. AB - Although the efficacy of tumor necrosis factor (TNF) antagonists for the treatment of rheumatoid arthritis (RA) has been established in randomized controlled trials (RCTs), safety concerns have emerged, particularly with regard to risk of infection, malignancy, and cardiovascular (CV) outcomes. Because of the shorter duration and limited number of patients enrolled in RCTs, evidence regarding the risk of rare adverse outcomes is frequently inconclusive. Longterm observational studies, as well as RCT meta-analyses, provide additional safety data. This review discusses the emerging evidence from observational registries on the risk of infection, malignancy, and CV outcomes associated with TNF antagonists. PMID- 17922667 TI - B cell biology and dysfunction in SLE. AB - Systemic lupus erythematosus (SLE) is a complex disease characterized by the production of autoantibodies and clinical involvement in multiple organ systems. The immunological events triggering the onset of clinical manifestations have not yet been fully defined, but a central role for B cells in the pathogenesis of this disease has been brought to the fore in the last several years by work performed at multiple laboratories in both mice and humans. B cell defects that have been defined include abnormal expression or function of key signaling molecules, dysregulation of cytokines with key B cell effects, and perturbations in B cell developmental subsets. Many of these defects may contribute to or be reflective of abnormalities in B cell tolerance. Both antibodydependent and antibody-independent mechanisms of B cells are important in SLE. Thus, autoantibodies contribute to autoimmunity by multiple mechanisms, including immunecomplex mediated type III hypersensitivity reactions and type II antibody dependent cytotoxicity, and by instructing innate immune cells to produce pathogenic cytokines, including IFNalpha, TNF, and IL-1. Autoantibody-independent B cell functions have been postulated to include antigen-presentation, T-cell activation and polarization, and dendritic cell (DC) modulation. Several of these functions are mediated by the ability of B cells to produce immuno-regulatory cytokines, chemokines, and lymphangiogenic growth factors and by their critical contribution to lymphoid tissue development and organization, including the development of ectopic tertiary lymphoid tissue. Given the large body of evidence implicating abnormalities in the B cell compartment in SLE,there has been a recent therapeutic focus on developing interventions that target the B cell compartment by multiple mechanisms. PMID- 17922669 TI - Neuropsychiatric lupus: clinical and imaging aspects. AB - Neuropsychiatric lupus (NPSLE) manifestations are common in adults and children and are associated with an increase in both morbidity and mortality. Cognitive dysfunction, when standardly assessed using sensitive neurocognitive instruments, is the most common NPSLE manifestation. The pathogenic etiologies of NPSLE manifestations are likely to be multifactorial and may involve autoantibody production, microangiopathy, intrathecal production of proinflammatory cytokines and athersclerosis. It is becoming more clear that the integrity of the blood brain-barrier is very important in SLE-related neuropathology. Brain imaging is an important tool that allows us to evaluate the living brain. Thus far, anatomic brain imaging has revealed abnormalities such as subcortical white matter lesions and cerebral atrophy, but these findings are non-specific. Methods that evaluate metabolic processes and other functional imaging techniques have more promise as surrogates for central nervous system damage. This article reviews the current literature on clinical and imaging aspects of NPSLE. PMID- 17922668 TI - Biomarker discovery in human SLE nephritis. AB - The treatment of systemic lupus erythematosus (SLE) nephritis, while effective, is associated with significant morbidity and mortality. These side effects could be mitigated if the onset, severity, and response of renal flare could be predicted, and therapy modified accordingly. In this review, an approach to derive prediction equations of SLE nephritis flare is discussed. Integral to generating such prediction equations is the identification of biomarkers of lupus nephritis that can serve as input variables for modeling flare. The use of urine as a source of SLE nephritis biomarkers is described, and the results of urine biomarker discovery studies using candidate and proteomic approaches are presented. PMID- 17922670 TI - Cutaneous lupus: insights into pathogenesis and disease classification. AB - Skin disease is the second most common manifestation in SLE patients, and a large number of patients have predominantly cutaneous lupus erythematosus. Experimental animal models suggest that modulation of immunologic factors can have a differential impact on the skin relative to the kidney, and therapeutic responses also suggest potential differences in the immunomodulation of skin relative to other organs affected in lupus. There have been recent insights into the etiology of cutaneous LE, including genetic and environmental factors. The growing understanding of the inflammatory cascade includes the role of UV-induction of pro-inflammatory cytokines, chemokines, and adhesion molecules. Apoptosis, necrosis, autoantibodies, plasmacytoid dendritic cells, T cells, B cells, and vascular changes all play a complex interactive role in the process of induction and maintenance of the lesions of cutaneous LE. Scientific efforts are beginning to clarify the pathophysiologic differences between subsets of cutaneous LE, but there are clearly many areas of investigation needed to elucidate the complex mechanisms that culminate in cutaneous LE. PMID- 17922672 TI - Vasculitis update, 2007. AB - Vasculitic syndromes are among the most complicated diseases to treat and manage. New medications and new ways of using old medications have provided us with new therapies to treat our patients. This review focuses on recent date that may have an impact on the way vasculitis is treated. PMID- 17922671 TI - Juvenile idiopathic arthritis: therapies in the 21st century. AB - Juvenile idiopathic arthritis (JIA) is an umbrella term for seven or more clinical patterns of arthritis of unknown cause in children. Until the mid-1980s, therapy for children, with what was then called juvenile rheumatoid arthritis in the United States and juvenile chronic arthritis (JRA) elsewhere, consisted primarily of a small repertoire of antiinflammatory drugs and corticosteroids. However, only a small percentage of children respond to NSAIDs (nonsteroidal antiinflammatory drugs) alone; almost all will respond to corticosteroids, but with the cost of unacceptable toxicities. Juvenile arthritis was often a crippling disease. The controlled trial that demonstrated methotrexate therapy was safe and effective in children was the major advance of that decade. With the burgeoning understanding of the immune system and the advent of biologic agents in the 21st century, pediatric rheumatologists now have many more therapies to offer patients, with the expectation that their disease will be controlled. This review will discuss current therapy and the approach to treatment of JIA. PMID- 17922673 TI - Hyperuricemia and gout: new insights into pathogenesis and treatment. AB - Over the past decade, significant advances have been made regarding the pathogenesis, clinical implications, and treatment of hyperuricemia. While physicians have understood for at least a century that uric acid causes gout, we are now beginning to address the question of why hyperuricemia exists and the mechanisms by which uric acid acts to stimulate inflammation. This review focuses on (1) previously unknown biological roles of uric acid; (2) why the loss of the uricase gene and resultant hyperuricemia may have provided an evolutionary advantage to primates and, in particular, to humans; (3) the molecular effects of uric acid on inflammatory cells; and (4) novel antihyperuricemic agents currently under study. PMID- 17922675 TI - The COX 2 selective inhibitors: what the newspapers have not told you. PMID- 17922674 TI - Osteoarthritis in 2007. AB - Osteoarthritis (OA) is often a progressive and disabling disease resulting from a combination of risk factors, including age, genetics, trauma, and knee alignment, as well as an imbalance of physiologic processes resulting in inflammatory cascades on a molecular level. The synovium, bone, and cartilage are each involved in the pathophysiological mechanisms that lead to progressive joint degeneration, and, thus, also serve as targets for therapies. Efforts to identify disease-modifying osteoarthritis drugs (DMOADs) have been hampered by several factors, but the focus has now shifted toward the validation of chemical and imaging biomarkers that should aid in DMOAD development. In this review, we summarize current pathological mechanisms occurring in the individual but interconnected compartments of OA joints, as well as discuss related therapeutic interventions that are currently available or on the horizon. PMID- 17922676 TI - DMOAD developments: present and future. AB - Osteoarthritis (OA), by far the most common form of arthritis, has a growing impact on health care. Progress in understanding its pathophysiological processes has led to the identification of promising therapeutic targets, with disease modifying osteoarthritis drugs (DMOADs) having the most potential. Numerous nonpharmaceutical measures and pharmacological interventions that slow the progression of the disease also have been developed. Several new classes of molecules that inhibit one or more OA pathophysiological processes have been discovered, and a number of these are under clinical evaluation to test their potential to alter the disease process in humans. Recent data from clinical trials have demonstrated that agents able to specifically block key disease mechanisms can effectively retard the progression of structural changes in knee OA patients. These studies are ushering the field into a new era in the development of DMOADs and, hence, the prospect of a cure for this disease. PMID- 17922677 TI - Impairment of the antioxidant properties of serum albumin in patients with diabetes: protective effects of metformin. AB - Free radical production is increased during diabetes. Serum albumin is a major antioxidant agent, and structural modification of albumin induced by glucose or free radicals impairs its antioxidant properties. Therefore the aim of the present study was to compare the antioxidant capacities and structural changes in albumin in patients with T2DM (Type 2 diabetes mellitus) treated with MET (metformin) or SU (sulfonylureas) and in healthy control subjects. Structural changes in albumin were studied by fluorescence quenching in the presence of acrylamide. Albumin thiols and fructosamines, reflecting oxidized and glycation induced changes in serum albumin respectively, were assessed. Structural changes in albumin were demonstrated by a significant decrease in fluorescence quenching in patients with T2DM, with patients treated with MET exhibiting a significant difference in the conformation of albumin compared with patients treated with SU. Oxidation, resulting in a significant decrease in thiol groups and plasma total antioxidant capacity, and glycation, associated with a significant increase in fructosamines, were both found when comparing healthy control subjects with patients with T2DM. When patients treated with MET were compared with those treated with SU, oxidative stress and glycation were found to be significantly lower in MET-treated patients. In conclusion, patients with T2DM have a decrease in the antioxidant properties of serum albumin which may aggravate oxidative stress and, thus, contribute to vascular and metabolic morbidities. Moreover, a significant protection of albumin was found in patients with T2DM treated with MET. PMID- 17922678 TI - An insight into the mechanism of inhibition of unusual bi-subunit topoisomerase I from Leishmania donovani by 3,3'-di-indolylmethane, a novel DNA topoisomerase I poison with a strong binding affinity to the enzyme. AB - DIM (3,3'-di-indolylmethane), an abundant dietary component of cruciferous vegetables, exhibits a wide spectrum of pharmacological properties. In the present study, we show that DIM is a potent inhibitor of Leishmania donovani topoisomerase I with an IC50 of 1.2 microM. Equilibrium dialysis shows that DIM binds strongly to the free enzyme with a binding constant of 9.73x10(-9) M. The binding affinity of DIM to the small subunit is 8.6-fold more than that of the large subunit of unusual LdTOP1LS (bi-subunit L. donovani topoisomerase I). DIM stabilizes topoisomerase I-DNA cleavage complexes in vitro and also in vivo. Like CPT (camptothecin), DIM inhibits the religation step when the drug was added to preformed topoisomerase I-DNA binary complex. Hence, DIM is similar to CPT with respect to its ability to form the topoisomerase I-mediated 'cleavable complexes' in vitro and in vivo. But unlike CPT, DIM interacts with both free enzyme and substrate DNA. Therefore DIM is a non-competitive class I inhibitor of topoisomerase I. DIM also inhibits the relaxation activity of the CPT-resistant mutant enzyme LdTOP1Delta39LS (N-terminal deletion of amino acids 1-39 of LdTOP1LS). The IC50 values of DIM in simultaneous and enzyme pre-incubation relaxation assays were 3.6 and 2.9 muM respectively, which are higher than that of wild-type topoisomerase I (LdTOP1LS), indicating that the affinity of DIM to LdTOP1Delta39LS is less than that for LdTOP1LS. This is the first report on DIM as an L. donovani topoisomerase I poison. Our study illuminates a new mode of action of enzyme inhibition by DIM that might be exploited for rational drug design in human leishmaniasis. PMID- 17922679 TI - Methionine sulfoxide reductases: selenoprotein forms and roles in antioxidant protein repair in mammals. AB - Msrs (methionine sulfoxide reductases), MsrA and MsrB, are repair enzymes that reduce methionine sulfoxide residues in oxidatively damaged proteins to methionine residues in a stereospecific manner. These enzymes protect cells from oxidative stress and have been implicated in delaying the aging process and progression of neurodegenerative diseases. In recent years, significant efforts have been made to explore the catalytic properties and physiological functions of these enzymes. In the current review, we present recent progress in this area, with the focus on mammalian MsrA and MsrBs including their roles in disease, evolution and function of selenoprotein forms of MsrA and MsrB, and the biochemistry of these enzymes. PMID- 17922681 TI - Expression of ADAMTS4 (aggrecanase-1) in human osteoarthritic cartilage. AB - A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)1, 4, 5, 8, 9 and 15, members of the ADAMTS gene family, have the ability to degrade a major cartilage proteoglycan, aggrecan, at the specific sites, and thus are called 'aggrecanases'. The expression of these ADAMTS species was examined in human osteoarthritic articular cartilage on reverse transcription-polymerase chain reaction. The results demonstrated the predominant expression of ADAMTS4 in osteoarthritic cartilage, while ADAMTS5 was constitutively expressed in osteoarthritic and normal cartilage. ADAMTS9 was expressed mainly in normal cartilage, whereas no or negligible expression of ADAMTS1, 8 and 15 was observed in either osteoarthritic or normal cartilage. In situ hybridization for ADAMTS4 indicated that chondrocytes in osteoarthritic cartilage expressed the mRNA. Two monoclonal antibodies to ADAMTS4 were developed, and immunolocalized ADAMTS4 to chondrocytes in the proteoglycan-depleted zones of osteoarthritic cartilage, showing a direct correlation with the Mankin scores. Immunoblotting indicated a major protein band of 58 kDa in the chondrocyte culture media and osteoarthritic cartilage tissue homogenates. These data demonstrate that among the six ADAMTS species, ADAMTS4 is mainly expressed in an active form in osteoarthritic cartilage, and suggest that ADAMTS4 may play an important role in the degradation of aggrecan in human osteoarthritic cartilage. PMID- 17922682 TI - Expression of connective tissue growth factor is in agreement with the expression of VEGF, VEGF-C, -D and associated with shorter survival in gastric cancer. AB - Connective tissue growth factor (CTGF) is believed to be a multifunctional signaling modulator involved in a wide variety of biological or pathological processes including carcinogenesis. The role of CTGF in gastric cancer (GC) has not been reported so far. In the present study the expression of CTGF, vascular endothelial growth factor (VEGF), VEGF-C and VEGF-D on immunohistochemistry in GC and the correlation between the expression of CTGF and VEGF, VEGF-C, VEGF-D were examined, along with the correlation between the expression of CTGF and clinicopathological parameters, as well as survival of the patients with GC. The expression of CTGF was significantly in agreement with expression of VEGF, VEGF-C and VEGF-D (kappa and P, respectively: 0.538, P < 0.001; 0.502, P < 0.001; 0.558, P < 0.001). High CTGF expression was significantly associated with lymph nodes metastasis (P = 0.038) and lower postoperative 5 year overall survival rates (23.9%) compared with those patients with low CTGF expression (48.4%, P = 0.0035). The present findings suggest that CTGF is a useful prognostic marker for GC. High CTGF expression is associated with the risk of lymph nodes metastasis and a poor survival time in GC. PMID- 17922683 TI - High-level expression of CD109 is frequently detected in lung squamous cell carcinomas. AB - CD109 is a glycosylphosphatidylinositol (GPI)-anchored cell surface protein, which is a member of the alpha2-macroglobulin/C3, C4, C5 family of thioester containing proteins. It has been reported that CD109 is expressed in a subset of hematopoietic cells, endothelial cells and several kinds of human tumors. Herein it is reported that the CD109 protein is preferentially expressed in lung squamous cell carcinomas compared with other types of lung carcinoma including adenocarcinomas, large cell carcinomas and small cell carcinomas. Immunohistochemical staining of surgically resected lung specimens using an anti CD109 antibody detected CD109 expression in basal cells of bronchial and bronchiolar epithelia and myoepithelial cells of bronchial secretary glands, but not in bronchial and bronchiolar apical epithelial cells and alveolar epithelial cells. Furthermore, the CD109 immunoreactivity was observed in squamous cell carcinomas at a high frequency compared with other types of lung carcinoma. Although the detailed function of CD109 protein is unclear, these results suggest that CD109 expression may play a role in the development of lung squamous cell carcinoma. PMID- 17922684 TI - Decreased expression of RIZ1 and its clinicopathological significance in epithelial ovarian carcinoma: correlation with epigenetic inactivation by aberrant DNA methylation. AB - The retinoblastoma protein-interacting zinc finger gene (RIZ1) is considered a tumor suppressor gene. The purpose of the present study was to examine the expression of RIZ1 and evaluate its clinicopathological significance in ovarian carcinoma. Immunohistochemistry and quantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed for RIZ1 and its clinicopathological significance was examined. DNA methylation status of RIZ1 was also studied. All (6/6) of the normal, 5/9 of benign, and 4/9 of borderline tissues were positive for RIZ1 protein. In ovarian cancer tissues 32.9% (54/164) were positive for RIZ1. Decreased expression of RIZ1 was significantly correlated with histological subtypes (P < 0.0001), high tumor grade (P = 0.0153) and advanced clinical stage (P = 0.0345), and high Ki67 index (P = 0.0117) but was not associated with the overall prognoses of the patients (P = 0.519). The presence of methylated band was detected in 2/9 cell lines, and 5/69 ovarian cancer tissues. Median values of relative RIZ1 expression in cell lines with methylation were significantly lower than those without methylation (P = 0.0404), and treatment of 5-aza 2'deoxycitidine resulted in demethylation and re-expression of RIZ1. Reduced expression of RIZ1 may play an important role in the pathogenesis and/or development of epithelial ovarian carcinoma, and is considered to be caused in part by aberrant DNA methylation. PMID- 17922686 TI - Intraductal carcinoma with complex fusion of tubular glands without macroscopic mucus in main pancreatic duct: dilemma in classification. AB - An 84-year-old man, who was being followed up after lobectomy for lung carcinoma, was referred for evaluation of a dilated main pancreatic duct (MPD) from the body to the tail. Endoscopic ultrasonography demonstrated a low-echo mass occupying the MPD from the body to the tail. Endoscopic retrograde pancreatography showed an occlusion of the MPD in the body, and brush cytology indicated malignant cells. Distal pancreatectomy was performed. Grossly, a white-yellow, irregular shaped solid mass without macroscopic mucus filled the lumen of the MPD. Histologically, the mass consisted of a complex fusion of tubular glands with atypical nuclei, which did not have intracellular mucus and oncocytic cytoplasm. The tumor mass showed abrupt transition to the normal epithelium. Immunohistochemically the tumor cells were partially positive for mucin 1 (MUC1) and MUC6, and negative for MUC2, MUC5AC, and lipase. Unfortunately the patient died of brain metastasis from lung carcinoma 15 months later. A review of reported cases of intraductal tubular tumors of the pancreas showed that the present case involved characteristics and immunohistochemical staining pattern similar to those of intraductal tubular carcinoma, although it might not be described as a typical intraductal tubular carcinoma under the existing Japanese rules. PMID- 17922685 TI - Increased expression of soluble form of vascular cell adhesion molecule-1 aggravates autoimmune arthritis in MRL-Fas(lpr) mice. AB - Vascular cell adhesion molecule-1 (VCAM-1, CD106) is important in leukocyte trafficking and its increased expression is associated with a number of chronic inflammatory diseases, including rheumatoid arthritis (RA). A soluble form of VCAM-1 (sVCAM-1) is generated by shedding of the membrane-bound molecule. The concentration of sVCAM-1 is increased in the sera of RA patients, but its pathological role has not been elucidated. The effect of sVCAM-1 relative to protection or aggravation of disease on the development of spontaneous arthritis was examined in an animal model of RA, namely MRL-Fas(lpr) mice (which display a disease resembling human RA), by generation of sVCAM-1 transgenic MRL-Fas(lpr) mice. Transgenic MRL-Fas(lpr) mice that expressed sVCAM-1 had higher incidence and increased severity of arthritis associated with higher levels of serum IgG rheumatoid factor compared with non-transgenic MRL-Fas(lpr) mice. These results suggest that sVCAM-1 plays an arthritogenic role in the development of inflammatory arthritis in MRL-Fas(lpr) mice and may present an important target for therapeutic strategy of RA. PMID- 17922687 TI - Amyloid deposition in primary pulmonary marginal zone B-cell lymphoma of mucosa associated lymphoid tissue. AB - A rare association between primary pulmonary marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma), and pulmonary immunoglobulin light chain (AL) amyloidosis is described in a 65-year-old woman suffering from rheumatoid arthritis (RA). All four nodules in the resected upper lobe of the lung had a similar histological appearance. They were composed of small-medium sized atypical lymphocytes. Centrocyte-like cells had lymphoepithelial lesions. Immunohistochemically, the tumor cells clonally expressed B-cell markers, and demonstrated clonal rearrangement of the immunoglobulin heavy chain gene on polymerase chain reaction. Based on these findings the diagnosis of primary pulmonary MALT lymphoma was made. In addition, uniform eosinophilic material deposition was identified randomly within the tumor. It was Congophilic and exhibited apple-green birefringence on polarizing microscopy, and remained unaffected by potassium permanganate digestion. Deposited material was immunoreactive to lambda light chain. It was concluded that this material was AL amyloid in primary pulmonary MALT lymphoma. Plasma cells with mRNA of lambda chain was found infiltrated along the border of amyloid deposition. Finally, it is speculated that primary pulmonary MALT lymphoma developing in an autoimmune setting, RA in the present case, is associated with overproduction and abnormal clearance of immunoglobulin by the tumor cells, resulting in AL amyloidosis within the tumor. PMID- 17922688 TI - Adrenal lipomas: incidental autopsy findings. AB - Two rare cases of lipomas of the adrenal gland are reported. In both cases the tumor was found incidentally at autopsy, in the course of the medicolegal investigation of death. The first case involved a small tumor of the left adrenal gland in a 70-year-old man who died by drowning. In the second case, a previously healthy 45-year-old man died suddenly. At autopsy a lipoma of the right adrenal gland was associated with a pheochromocytoma in the contralateral gland. PMID- 17922689 TI - Juxtaoral organ of Chievitz, radiologically suspicious for invasion of lingual squamous cell carcinoma. PMID- 17922691 TI - Microparticles and exosomes: impact on normal and complicated pregnancy. AB - Eukaryotic cells release vesicles into their environment by membrane shedding (ectosomes or microparticles) and secretion (exosomes). Microparticles and exosomes occur commonly in vitro and in vivo. The occurrence, composition and function(s) of these vesicles change during disease (progression). During the last decade, the scientific and clinical interest increased tremendously. Evidence is accumulating that microparticles and exosomes may be of pathophysiological relevance in autoimmune, cardiovascular and thromboembolic diseases, as well as inflammatory and infectious disorders. In this review, we will summarize the discovery, biology, structure and function of microparticles and exosomes, and discuss their (patho-) physiological role during normal and complicated pregnancy. PMID- 17922692 TI - Polymorphisms in the interleukin-12/18 genes and recurrent spontaneous abortion. AB - PROBLEM: Interleukin (IL) IL-12/IL-18 are involved in uterine NK cells control of uterine vascular development. Polymorphisms in the IL-12/IL-18 genes could modify the cytokine balance, which might result in an increased susceptibility to recurrent spontaneous abortion (RSA). METHOD OF STUDY: A case-control study was conducted to determine the association between the IL12 (I/D) and IL18 (-607C>A, 137G>C) gene polymorphisms and the risk of RSA in 125 women with RSA and in 136 controls. RESULTS: The frequencies of DD, ID, II for IL-12 were, 25.6%, 52.8% and 21.6% respectively, in patients versus 21.3%, 51.5% and 27.2% respectively in controls; the frequencies of CC, CA, AA genotypes for IL-18 (-607) were, 34.4%, 54.4% and 11.2% respectively in patients versus 30.1%, 58.1% and 11.8% respectively in controls; the frequencies of GG, GC, CC genotypes for IL-18(-137) were 47.2%, 43.2% and 9.6% respectively in patients and 45.6%, 46.3% and 8.1% respectively in controls. CONCLUSION: IL-12B and IL-18 promoter gene polymorphisms were not associated with RSA in our women. PMID- 17922693 TI - Increased natural killer cell cytotoxicity and IL-2 production in recurrent spontaneous abortion. AB - PROBLEM: To determine whether natural killer (NK) cells cytotoxicity in peripheral blood is altered in patients with a history of recurrent spontaneous abortion (RSA); also, if there is any correlation between cytokine production and NK cytotoxicity. METHOD OF STUDY: In this case-control study, 21 patients with RSA within 24 hr of the last abortion (group I), and 32 pregnants with no history of abortion (group II) were surveyed. NK cell cytotoxicity was evaluated by flow cytometry, and IL-2, IL-10, transforming growth factor beta1 were measured in cell culture supernatant by ELISA method. RESULTS: Group I showed higher NK cytotoxicity than group II at all of effector to target (E:T) ratios (P < or = 0.045). The correlation between production of IL-2 and NK cytotoxicity was positively significant (R = 0.350, P = 0.001). Group I had significantly higher levels of IL-2 than group II (P = 0.001). In group II, the production of IL-10 by peripheral blood mononuclear cells was higher than group I (P = 0.002). CONCLUSION: Increased NK cell cytotoxicity and high level of IL-2 may be considered as a risk factor for RSA. PMID- 17922694 TI - Distribution and spatiotemporal relationship of activin a and follistatin in mouse decidual and placental tissue. AB - Problem Cells responsible for the synthesis of follistatin and activin A in the pregnant mouse endometrium have not been characterized. Method of study Immunocytochemistry was used to determine the distribution of follistatin and activin A in the pregnant mouse uterus. Results Follistatin was detected in the endometrium prior to decidualization and embryo implantation. Follistatin was not seen in fully decidualized cells, being restricted to non-decidualized fibroblasts and cells in the process of decidualization. In contrast, activin A was detected exclusively in mature antimesometrial decidual cells during involution. After day eleven of pregnancy, both substances were identified in the extracellular matrix of the spongiotrophoblast. Conclusion As previously described for decidual prolactin-related protein and the proteoglycan perlecan, follistatin and activin A were detected in the extracellular matrix of the spongiotrophoblast, suggesting that this region acts as reservoir for these growth factors in the mouse placenta. PMID- 17922695 TI - Female sex steroid hormones modify some regulatory properties of monocyte-derived dendritic cells. AB - PROBLEM: Monocyte-derived dendritic cells (mDCs) are present in human decidua during the first month of pregnancy where they experience the effect of the increased concentration of progesterone (Pg) and estradiol (Estr). The aim of our study was to assess the effect of these reproductive hormones on the immunomodulatory role of mDCs. METHODS OF STUDY: The mDCs cultured in vitro in the presence of sex steroid hormones and characterized for their apoptosis and secretion of cytokines by flow cytometry, enzyme linked immunosorbent assay and proteome profiler kit. RESULTS: Female sex steroid hormones enhance the life cycle of mDCs, thus increasing the maturation and apoptosis, they also increase the production of interleukin (IL)-10 and IL-27 but only Pg increases the production of IL-13 and down regulates the secretion of IL-23. CONCLUSION: The Pg and Estr participate in the regulation of the immune response during pregnancy by influencing apoptosis and the cytokine profile of the mDCs. PMID- 17922696 TI - The effects of platelet-activating factor on the secretion of interleukin-8 and growth-regulated oncogene alpha in human immortalized granulosa cell line (GC1a). AB - PROBLEM: To investigate the role of platelet-activating factor (PAF) in human ovulation, we studied the regulation of interleukin (IL)-8 and growth-regulated oncogene (GRO) alpha in cultured human immortalized granulosa cell line (GC1a). METHOD OF STUDY: GC1a was cultured in serum-free medium, and incubated with carbamyl-PAF (C-PAF) and/or PAF receptor antagonist (WEB 2086). The supernatants were collected, and IL-8 and GRO alpha were measured by enzyme-linked immunosorbent assay. RESULTS: After treatment with C-PAF, the levels of IL-8 and GROalpha increased in a time-dependent manner. The levels of IL-8 and GROalpha were significantly increased after treatment with C-PAF in a dose-dependent manner. However, the levels of IL-8 and GROalpha were significantly decreased by treatment with C-PAF and with increasing concentrations of WEB 2086. CONCLUSION: Our data indicated that IL-8 and GROalpha were regulated by C-PAF. The results suggested that PAF may play an important role in human pre-ovulatory processes involving IL-8 and GROalpha production. PMID- 17922697 TI - Relationship between human tumor-associated antigen RCAS1 and gestational diabetes mellitus. AB - PROBLEM: The human tumor-associated receptor-binding cancer antigen expressed on SiSo cells (RCAS1) is considered to play a role in the inhibition of the maternal immune response during pregnancy. The aim of our study was to investigate the expression of RCAS1 protein in the placenta and to compare its concentration in maternal and cord blood sera between normal pregnancies and pregnancies complicated by gestational diabetes mellitus (GDM). METHOD OF STUDY: Samples were obtained from women with GDM (n = 8), women with type 1 DM (n = 5), and healthy term controls (n = 27) after delivery. Placentas were studied by immunohistochemistry, and real-time polymerase chain reaction. For assessment of RCAS1 concentrations in maternal and cord blood sera, enzyme-linked immunosorbent assay was performed. RESULTS: The RCAS1 protein mRNA expression in the placentas of patients with GDM was significantly lower than that in the controls (P = 0.005). The maternal blood RCAS1 protein concentration of the GDM cases was also significantly lower than that in the controls (P = 0.0411), whereas the cord blood RCAS1 protein concentration was significantly higher in the GDM and type 1 DM groups than in the controls (P = 0.0311 and P = 0.0192, respectively). CONCLUSION: The present results suggest that RCAS1 protein might have an important role in the development of GDM. PMID- 17922698 TI - Decline in number of elevated blood CD3(+) CD56(+) NKT cells in response to intravenous immunoglobulin treatment correlates with successful pregnancy. AB - PROBLEM: Patients with elevated blood natural killer (NK) cells may be offered intravenous immunoglobulin (IVIG) treatment, but there is controversy about the utility of blood NK cell testing. Human CD56(+) NK cells include several subpopulations that include the putatively cytotoxic CD56(+) CD16(+) subset. In mouse models of pregnant failure, NKT cells appear to be important. However, a mouse model may only be pertinent to a subset of patients, as recurrent pregnancy failure is a heterogenous group. METHOD OF STUDY: An ethics-approved observational study was done to observe the effect of treatment on total blood lymphoid cells, and subsets of CD56(+) blood lymphocytes including CD56(+) CD3(+) NKT cells determined by flow cytometry, and to correlate with pregnancy outcome. Fifteen fertile women with a history of successful pregnancy and thirty-one women suffering from repeated implantation failure or recurrent spontaneous abortion provided serial blood samples during one menstrual cycle or prior to and during treatment. IVIG was administered to the latter group with or without heparin/aspirin. RESULTS: Eight of thirty infertile women presented with high numbers of CD56(+) CD3(+) NKT cells, which declined after treatment with IVIG. The elevated NKT cell group with or without concomitant autoimmunity achieved a significantly higher successful pregnancy rate over the course of the study, as compared to women with average numbers of NKT cells and no evidence of autoimmunity (P = 0.018). Elevated NKT levels alone was an independent predictor of success on treatment (P = 0.003). CONCLUSION: Elevated NKT cells in recurrent pregnancy loss or implantation failure can be ameliorated with IVIG treatment, and result in successful pregnancy. Assay of NKT cell numbers may identify patients who are more likely to benefit from IVIG therapy and merits further examination in randomized phase II studies. PMID- 17922699 TI - The placental barrier in allogenic immune conflict in spontaneous early abortions: immunohistochemical and morphological study. AB - PROBLEM: Morphologic changes in the placental barrier in spontaneous early abortions under the maternal-embryonic immune conflict, and the role of maternal immunoglobulins (Igs) in these changes. MATERIALS AND METHODS: We examined chorionic villi and other tissues obtained from 54 aborts between weeks 3.5 and 8 of pregnancy. Material was divided into two groups. Group 1 (control) contained 15 medically recommended and spontaneous early aborts with no signs of inflammations or pathologic immune processes. Group 2 contained 39 spontaneous early aborts with acute chorionic villitis. Immunohistochemical and morphometric methods were used to study the Igs, different types of immunocompetent cells, and apoptosis-related components of the placental barrier. RESULTS: Acute villitis was found to be characterized by the destruction of all components of the chorionic villi, thrombovasculitis with apoptosis of the endothelium of capillaries and erythroblasts, mucous swelling of the basal membrane, and coagulation of the blood proteins. Due to destruction of the capillaries, the number of avasculate villi increased, and the average number of capillaries per villus decreased. The extremely high number of phagolysosomes with IgG and IgA in the villous monocytes in the group 2 indicates an increase in the phagocytic activity of monocytes against maternal Igs and may reflect the presence of mother embryo immune conflict. Apoptosis of monocytes and a high number of promonocytes were seen accompanied by a high concentration of p53 protein. A large disturbance in the trophoblast occurred with disappearance of bcl-2 and the appearance of Fas ligand. CONCLUSION: Massive destruction of maternal Igs in embryonic monocytes and acute villitis in the placental barrier are manifested during the mother embryo immune conflict, and this may be one of the reasons of spontaneous early abortions. PMID- 17922700 TI - The anatomy of predator-prey dynamics in a changing climate. AB - Humans are increasingly influencing global climate and regional predator assemblages, yet a mechanistic understanding of how climate and predation interact to affect fluctuations in prey populations is currently lacking. Here we develop a modelling framework to explore the effects of different predation strategies on the response of age-structured prey populations to a changing climate. We show that predation acts in opposition to temporal correlation in climatic conditions to suppress prey population fluctuations. Ambush predators such as lions are shown to be more effective at suppressing fluctuations in their prey than cursorial predators such as wolves, which chase down prey over long distances, because they are more effective predators on prime-aged adults. We model climate as a Markov process and explore the consequences of future changes in climatic autocorrelation for population dynamics. We show that the presence of healthy predator populations will be particularly important in dampening prey population fluctuations if temporal correlation in climatic conditions increases in the future. PMID- 17922701 TI - Climate change can alter competitive relationships between resident and migratory birds. AB - Climate change could affect resource competition between resident and migratory bird species by changing the interval between their onsets of breeding or by altering their population densities. We studied interspecific nest-hole competition between resident great tits and migrant pied flycatchers in South Western Finland over the past five decades (1953-2005). We found that appearance of fatal take-over trials, the cases where a pied flycatcher tried to take over a great tit nest but was killed by the tit, increased with a reduced interspecific laying date interval and with increasing densities of both tits and flycatchers. The probability of pied flycatchers taking over great tit nests increased with the density of pied flycatchers. Laying dates of the great tit and pied flycatcher are affected by the temperatures of different time periods, and divergent changes in these temperatures could consequently modify their competitive interactions. Densities are a result of reproductive success and survival, which can be affected by separate climatic factors in the resident great tit and trans-Saharan migrant pied flycatcher. On these bases we conclude that climate change has a great potential to alter the competitive balance between these two species. PMID- 17922702 TI - Climatic variation and the distribution of an amphibian polyploid complex. AB - The establishment of polyploid populations involves the persistence and growth of the polyploid in the presence of the progenitor species. Although there have been a number of animal polyploid species documented, relatively few inquiries have been made into the large-scale mechanisms of polyploid establishment in animal groups. Herein we investigate the influence of regional climatic conditions on the distributional patterns of a diploid-tetraploid species pair of gray treefrogs, Hyla chrysoscelis and H. versicolor (Anura: Hylidae) in the mid Atlantic region of eastern North America. Calling surveys at breeding sites were used to document the distribution of each species. Twelve climatic models and one elevation model were generated to predict climatic and elevation values for gray treefrog breeding sites. A canonical analysis of discriminants was used to describe relationships between climatic variables, elevation and the distribution of H. chrysoscelis and H. versicolor. There was a strong correlation between several climatic variables, elevation and the distribution of the gray treefrog complex. Specifically, the tetraploid species almost exclusively occupied areas of higher elevation, where climatic conditions were relatively severe (colder, drier, greater annual variation). In contrast, the diploid species was restricted to lower elevations, where climatic conditions were warmer, wetter and exhibited less annual variation. Clusters of syntopic sites were associated with areas of high variation in annual temperature and precipitation during the breeding season. Our data suggest that large-scale climatic conditions have played a role in the establishment of the polyploid H. versicolor in at least some portions of its range. The occurrence of the polyploid and absence of the progenitor in colder, drier and more varied environments suggests the polyploid may possess a tolerance of severe environmental conditions that is not possessed by the diploid progenitor. Our findings support the hypothesis that increased tolerance to severe environmental conditions is a plausible mechanism of polyploid establishment. PMID- 17922703 TI - Stream flow, salmon and beaver dams: roles in the structuring of stream fish communities within an anadromous salmon dominated stream. AB - The current paradigm of fish community distribution is one of a downstream increase in species richness by addition, but this concept is based on a small number of streams from the mid-west and southern United States, which are dominated by cyprinids. Further, the measure of species richness traditionally used, without including evenness, may not be providing an accurate reflection of the fish community. We hypothesize that in streams dominated by anadromous salmonids, fish community diversity will be affected by the presence of the anadromous species, and therefore be influenced by those factors affecting the salmonid population. Catamaran Brook, New Brunswick, Canada, provides a long-term data set to evaluate fish community diversity upstream and downstream of an obstruction (North American beaver Castor canadensis dam complex), which affects distribution of Atlantic salmon Salmo salar. The Shannon Weiner diversity index and community evenness were calculated for sample sites distributed throughout the brook and over 15 years. Fish community diversity was greatest upstream of the beaver dams and in the absence of Atlantic salmon. The salmon appear to depress the evenness of the community but do not affect species richness. The community upstream of the beaver dams changes due to replacement of slimy sculpin Cottus cognatus by salmon, rather than addition, when access is provided. Within Catamaran Brook, location of beaver dams and autumn streamflow interact to govern adult Atlantic salmon spawner distribution, which then dictates juvenile production and effects on fish community. These communities in an anadromous Atlantic salmon dominated stream do not follow the species richness gradient pattern shown in cyprinid-dominated streams and an alternative model for stream fish community distribution in streams dominated by anadromous salmonids is presented. This alternative model suggests that community distribution may be a function of semipermeable obstructions, streamflow and the distribution of the anadromous species affecting resident stream fish species richness, evenness, biomass and production. PMID- 17922704 TI - Does interference competition with wolves limit the distribution and abundance of coyotes? AB - Interference competition with wolves Canis lupus is hypothesized to limit the distribution and abundance of coyotes Canis latrans, and the extirpation of wolves is often invoked to explain the expansion in coyote range throughout much of North America. We used spatial, seasonal and temporal heterogeneity in wolf distribution and abundance to test the hypothesis that interference competition with wolves limits the distribution and abundance of coyotes. From August 2001 to August 2004, we gathered data on cause-specific mortality and survival rates of coyotes captured at wolf-free and wolf-abundant sites in Grand Teton National Park (GTNP), Wyoming, USA, to determine whether mortality due to wolves is sufficient to reduce coyote densities. We examined whether spatial segregation limits the local distribution of coyotes by evaluating home-range overlap between resident coyotes and wolves, and by contrasting dispersal rates of transient coyotes captured in wolf-free and wolf-abundant areas. Finally, we analysed data on population densities of both species at three study areas across the Greater Yellowstone Ecosystem (GYE) to determine whether an inverse relationship exists between coyote and wolf densities. Although coyotes were the numerically dominant predator, across the GYE, densities varied spatially and temporally in accordance with wolf abundance. Mean coyote densities were 33% lower at wolf-abundant sites in GTNP, and densities declined 39% in Yellowstone National Park following wolf reintroduction. A strong negative relationship between coyote and wolf densities (beta = -3.988, P < 0.005, r(2) = 0.54, n = 16), both within and across study sites, supports the hypothesis that competition with wolves limits coyote populations. Overall mortality of coyotes resulting from wolf predation was low, but wolves were responsible for 56% of transient coyote deaths (n = 5). In addition, dispersal rates of transient coyotes captured at wolf-abundant sites were 117% higher than for transients captured in wolf-free areas. Our results support the hypothesis that coyote abundance is limited by competition with wolves, and suggest that differential effects on survival and dispersal rates of transient coyotes are important mechanisms by which wolves reduce coyote densities. PMID- 17922705 TI - Geographical ecology of Paraguayan bats: spatial integration and metacommunity structure of interacting assemblages. AB - We examined the relative contributions of regional spatial characteristics and local environmental conditions in determining Paraguayan bat species composition. We used a suite of full and partial redundancy analyses to estimate four additive partitions of variance in bat species composition: (a) unexplained variation, (b) that explained purely by spatial characteristics, (c) that explained purely by local environmental conditions and (d) that explained jointly by space and environment. The spatial component to bat species composition was greater than the environmental component and both pure spatial and pure environmental characteristics accounted for significant amounts of variation in bat species composition. Results from variance decomposition suggest that the mass effects model describes metacommunity structure of Paraguayan bats better than species sorting or neutral models. Such mass effects may potentially be general for bats and could explain the inability of purely local factors to fully account for bat community organization. Mass effects also have substantial conservation implications because rescue effects may enhance the persistence of mobile species in fragmented landscapes with relatively few protected sites. PMID- 17922706 TI - Experimental evidence for the effects of dung beetle functional group richness and composition on ecosystem function in a tropical forest. AB - Much of the literature on the relationship between species richness or functional group richness and measures of ecosystem function focuses on a restricted set of ecosystem function measures and taxonomic groups. Few such studies have been carried out under realistic levels of diversity in the field, particularly in high diversity ecosystems such as tropical forests. We used exclusion experiments to study the effects of dung beetle functional group richness and composition on two interlinked and functionally important ecological processes, dung removal and secondary seed dispersal, in evergreen tropical forest in Sabah, Malaysian Borneo. Overall, both dung and seed removal increased with dung beetle functional group richness. However, levels of ecosystem functioning were idiosyncratic depending on the identity of the functional groups present, indicating an important role for functional group composition. There was no evidence for interference or competition among functional groups. We found strong evidence for overyielding and transgressive overyielding, suggesting complementarity or facilitation among functional groups. Not all mixtures showed transgressive overyielding, so that complementarity was restricted to particular functional group combinations. Beetles in a single functional group (large nocturnal tunnellers) had a disproportionate influence on measures of ecosystem function: in their absence dung removal is reduced by approximately 75%. However, a full complement of functional groups is required to maximize ecosystem functioning. This study highlights the importance of both functional group identity and species composition in determining the ecosystem consequences of extinctions or altered patterns in the relative abundance of species. PMID- 17922707 TI - The significance of ratios of detritus types and micro-organism productivity to competitive interactions between aquatic insect detritivores. AB - Investigations of competitive interactions emphasize non-detrital resources, even though detritus is a major component of most food webs. Studies of competing species focus usually on single resource types, although consumers in nature are likely to encounter mixtures of resource types that may affect whether competition results in exclusion or coexistence. The invasive mosquito Aedes albopictus is capable of excluding the native mosquito Ochlerotatus triseriatus in competition for single detritus types in laboratory and field microcosms. In this study, we used nine ratios of two detritus types (animal and leaf) common in natural containers to test whether detritus ratios affect the outcome of competition. Under intraspecific and interspecific competition, A. albopictus attained higher survival and estimated population growth rate than did O. triseriatus. Unlike past studies, both species had positive growth and high adult survival, with little evidence of competitive effects, under one resource ratio (10:1 ratio of leaf : animal detritus) regardless of mosquito densities, suggesting potential coexistence. Path analysis showed that densities of larvae had negative effects on population growth for O. triseriatus but not for A. albopictus, indicating competitive superiority of A. albopictus. Population growth of both species was affected strongly by the direct paths from animal (positive) and leaf (negative) detritus, and the indirect effect of leaf detritus via bacterial production (positive). Field sampling established that detritus entered real tree holes in ratios similar to those in our experiment, suggesting that natural variation in detritus ratios may influence local coexistence of these species. Seasonal variation in ratios of plant and animal detritus indicated that temporal as well as spatial variation in inputs may be important for potential coexistence. PMID- 17922708 TI - Short-term transformation of matrix into hospitable habitat facilitates gene flow and mitigates fragmentation. AB - Habitat fragmentation has major implications for demography and genetic structure of natural plant and animal populations as small and isolated populations are more prone to extinction. Therefore, many recent studies focus on spatial fragmentation. However, the temporal configuration of suitable habitat may also influence dispersal and gene flow in fragmented landscapes. We hypothesize that short-term switching of inhospitable matrix areas into suitable habitat can mitigate effects of spatial fragmentation in natural and seminatural ecosystems. To test our hypothesis, we investigated the hairy-footed gerbil (Gerbillurus paeba, Smith 1836), a ground-dwelling rodent, in fragmented Kalahari savannah areas. Here, rare events of high above mean annual rainfall suggest short-term matrix suitability. During the field survey in 'matrix' areas in the Kalahari (shrub encroachment by heavy grazing) we never observed the hairy-footed gerbil in years of average rainfall, but observed mass occurrences of this species during rare events of exceptionally high rainfall. In a second step, we developed an agent-based model simulating subpopulations in two neighbouring habitats and the separating matrix. Our mechanistic model reproduces the mass occurrences as observed in the field and thus suggests the possibly underlying processes. In particular, the temporary improvement in matrix quality allows reproduction in the matrix, thereby causing a substantial increase in population size. The model demonstrates further how the environmental trigger (rainfall) impacts genetic connectivity of two separated subpopulations. We identified seasonality as a driver of fragmentation but stochasticity leading to higher connectivity. We found that our concept of temporal fragmentation can be applied to numerous other fragmented populations in various ecological systems and provide examples from recent literature. We conclude that temporal aspects of fragmentation must be considered in both ecological research and conservation management. PMID- 17922709 TI - Behavioural syndromes differ predictably between 12 populations of three-spined stickleback. AB - Animals often differ in suites of correlated behaviours, comparable with how humans differ in personality. Constraints on the architecture of behaviour have been invoked to explain why such 'behavioural syndromes' exist. From an adaptationist viewpoint, however, behavioural syndromes should evolve only in those populations where natural selection has favoured such trait covariance, and they should therefore exist only in particular types of population. A comparative approach was used to examine this prediction of the adaptive hypothesis. We measured behavioural correlations in 12 different populations of three-spined stickleback (Gasterosteus aculeatus) and assessed whether they indeed varied consistently according to the selective environment, where population was unit of analysis. For a sample of fry from each population, we measured five different behaviours within the categories of (i) aggression (towards conspecifics); (ii) general activity; and (iii) exploration-avoidance (of novel foods, novel environments and altered environments). We show that behavioural syndromes are not always the same in different types of stickleback population: the often documented syndrome between aggressiveness, activity and exploratory behaviour existed only in large ponds where piscivorous predators were present. In small ponds where predators were absent, these behaviours were not (or only weakly) associated. Our findings imply that population variation in behavioural syndromes does not result from stochastic evolutionary processes, but may result instead from adaptive evolution of behaviour favouring what should prove to be optimal trait combinations. PMID- 17922710 TI - Evidence for a trade-off between early growth and tooth wear in Svalbard reindeer. AB - Ruminants depend on efficient physical degradation of forage through chewing to increase the surface area of the food particles presented to the microflora. Fossil evidence suggests that increased molar height is an adaptation for wear tolerance in dry ecosystems with sparse vegetation, but no study has shown selection pressure for hypsodonty in contemporary ruminants. We explored the relationships between particle size in rumen, tooth wear (scanned molar occlusal topography), age and body mass of female Svalbard reindeer living in an arctic desert at 78 degrees latitude on Svalbard. We predicted that (H1) if the rumen particle size is determined mainly by constraints due to tooth wear, and if tooth wear is mainly a function of age, average particle size in rumen should increase with age. From allometric relations it is known that larger individuals can survive on a lower-quality diet, we therefore predicted (H2) larger particle sizes with increases in (ln) body mass, irrespective of age and wear. Lastly, if there is a trade-off between growth and tooth wear in dry ecosystems (a selection pressure for hypsodonty), we predicted (H3) that teeth of heavier animals should be more worn than those of lighter animals of the same age. The proportion of small particles (<1.0 mm) decreased rapidly with increasing age (consistent with H1). Heavier females within an age class had more worn teeth (consistent with H3) than lighter ones. A close-to-isometric relationship between particle size and body mass suggested that heavier animals partly compensated for reduced tooth efficiency by chewing more. We provide the first evidence of a trade-off between fast early growth and wear (a somatic cost) of a senescence-related trait--the structure and height of the molar--in a wild ruminant inhabiting an arctic desert where selection pressure for increased tooth height is expected. This suggests that foraging conditions are more extreme than the environment in which the species originally evolved. PMID- 17922711 TI - Population dynamics in a long-lived seabird: I. Impact of breeding activity on survival and breeding probability in unbanded king penguins. AB - Understanding the trade-off between current reproductive effort, future survival and future breeding attempts is crucial for demographic analyses and life history studies. We investigated this trade-off in a population of king penguins (Aptenodytes patagonicus) marked individually with transponders using multistate capture-recapture models. This colonial seabird species has a low annual proportion of non-breeders (13%), despite a breeding cycle which lasts over 1 year. To draw inferences about the consequences of non-breeding, we tested for an effect of reproductive activity on survival and on the probability of subsequent breeding. We found that birds non-breeding in year t show the same survival rate as breeders (two-states analysis: breeding and non-breeding). However, breeders had a lower probability of breeding again the following year. This negative phenotypic correlation suggests the existence of reproductive costs affecting future breeding probability, but it might also be strengthened by late arrival for courtship in year t. A three-state analysis including breeding success revealed that failed breeders in year t have a lower probability to reproduce successfully in year t + 1 than non-breeders in year t, providing some evidence for the existence of reproductive costs. Moreover, successful breeders showed higher survival probability. This positive phenotypic correlation between current reproduction and subsequent survival supports the hypothesis of an heterogeneity in individual quality. Males breeding in year t had a lower probability to breed again in year t + 1 than females, suggesting higher reproductive costs for this sex. Such additional costs might be due to higher male parental investment in the final phase of chick-rearing, which also delays the arrival of males in year t + 1, and decreases their breeding probability. Our study is the first to explore the breeding biology and the demography of penguins without the disturbance of flipper-bands. PMID- 17922712 TI - Male reproductive senescence: the price of immune-induced oxidative damage on sexual attractiveness in the blue-footed booby. AB - In animals, male reproduction is commonly a function of sexual attractiveness, based on the expression of sexually dimorphic traits that advertise genuinely the male's quality. Male performance may decline with age because physiological functions underlying sexual attractiveness may be affected by senescence. Here we show that a sexual signal (foot colour) declines with age, due probably to the deleterious effects of oxidative damage. We found that in the blue-footed booby Sula nebouxii foot colour during courtship was less attractive in senescent than in middle-aged males. In addition, we increased reactive oxygen species experimentally by immunizing males with lipopolysaccharide, a bacterial cell wall component that induces marked oxidative stress in animals. The immune system activation induced greater lipid peroxidation and invoked changes on colour expression (less attractive), particularly in senescent males. These results support the idea that oxidative stress affects reproductive senescence, and suggest that oxidative damage might be a proximal mechanism underlying age reproductive patterns in long-lived animals. PMID- 17922713 TI - Estimating expenditure on male and female offspring in a sexually size-dimorphic bird: a comparison of different methods. AB - The parents of sexually size-dimorphic offspring are often assumed to invest more resources producing individuals of the larger sex. A range of different methods have been employed to estimate relative expenditure on the sexes, including quantifying sex-specific offspring growth, food intake, energy expenditure and energy intake, in addition to measures of parental food provisioning and energy expenditure. These methods all have the potential to provide useful estimates of relative investment, but each has particular problems of interpretation, and few studies have compared the estimates derived concurrently from more than two of these measures. In this study we compared these surrogate measures of parental investment in the brown songlark Cinclorhamphus cruralis, which exhibits one of the most extreme cases of sexual size dimorphism among birds. At 10 days of age we found that male chicks, on average, were 49% heavier, received 42% more prey items, expended 44% more energy and ingested 50% more metabolizable energy than their sisters. Furthermore, we created, experimentally, both all-male and all female broods of 10-day-old chicks and found that mothers delivered 43% more prey items and expended 27% more energy when provisioning all-male broods, providing the first direct evidence for a change in parental energy expenditure in relation to brood sex ratio. These data reveal remarkable agreement between these estimates of investment and suggest that all may provide quantitatively useful information on sex allocation. However, the lower variance associated with estimates of relative mass and energy intake suggest that these methods may be of greater utility, although this may primarily reflect the shorter period over which our provisioning data were collected. PMID- 17922714 TI - Age-specific reproductive success in a long-lived bird: do older parents resist stress better? AB - In many vertebrates, reproductive performance increases with advancing age but mechanisms involved in such a pattern remain poorly studied. One potential mechanism may be the hormonal stress response, which shifts energy investment away from reproduction and redirects it towards survival. In birds, this stress response is achieved through a release of corticosterone and is also accompanied by a decrease in circulating prolactin, a hormone involved widely in regulating parental cares. It has been predicted that, when the value of the current reproduction is high relative to the value of future reproduction and survival, as it is expected to be in older adults, the stress response should be attenuated to ensure that reproduction is not inhibited. We tested this hypothesis by measuring the corticosterone and prolactin responses of known-age (8-36 years old) incubating snow petrels (Pagadroma nivea) to a standardized capture/handling stress protocol. We also investigated whether an attenuation of the stress responses will correlate with a lower occurrence of egg neglect, a frequently observed behaviour in snow petrels. The probability of successfully fledging a chick increased from 6 years to 12 years before stabilizing after 12 years of age. Corticosterone response to stress was unaffected by age. Prolactin response to stress, however, was influenced clearly by age: in both sexes older breeders had higher stress-induced prolactin levels than younger ones. This was due to an increasing attenuation of the prolactin response to stress with advancing age in females, and in males this was due to a probably higher intrinsic capacity of older males to secrete prolactin. Moreover, higher stress-induced prolactin levels were correlated with a lower probability of neglecting the egg. In young breeders, the combination of a robust corticosterone increase with a lower ability to maintain prolactin secretion during acute stress is probably one of the functional causes of their lower incubation commitment. We suggest that the ability to maintain a threshold level of prolactin during a stressful situation may be an important physiological mechanism involved in the improvement of reproductive performance with advancing age in long-lived birds. PMID- 17922715 TI - Female red squirrels fit Williams' hypothesis of increasing reproductive effort with increasing age. AB - Williams predicted that reproductive effort should increase as individuals age and their reproductive value declines. This simple prediction has proven difficult to test because conventional measures of energy expenditure on reproduction may not be a true reflection of reproductive effort. We investigated age-specific variation in female reproductive effort in a stable population of North American red squirrels where energy expenditure on reproduction is likely to reflect actual reproductive effort. We used seven measures of reproductive effort spanning conception to offspring weaning. We found that females completed growth by age 3 and that reproductive value decreased after this age likely because of reproductive and survival senescence. We therefore, predicted that reproductive effort would increase from age 3 onwards. The probability of breeding, litter mass at weaning, and likelihood of territory bequeathal were all lower for 1- and 2-year-old females than for females older than 3 years, the age at which growth is completed. That growing females are faced with additional energetic requirements might account for their lower allocation to reproduction as compared with older females. The probability of attempting a second reproduction within the same breeding season and the propensity to bequeath the territory to juveniles increased from 3 years of age onwards, indicating an increase in reproductive effort with age. We think this increase in reproductive effort is an adaptive response of females to declining reproductive values when ageing, thereby supporting Williams' prediction. PMID- 17922716 TI - Evidence for a cost of immunity when the crustacean Daphnia magna is exposed to the bacterial pathogen Pasteuria ramosa. AB - The deployment of the immune system has the obvious potential to ameliorate infection outcomes, but immune responses can also harm hosts by either damaging host tissues or monopolizing resources, leading to enhanced mortality. To gain insight into such a 'cost of immunity' when the crustacean Daphnia magna is challenged with the bacterium Pasteuria ramosa, we measured survivorship among hosts that resisted infection following exposure to various strains and doses of the parasite. In the first of two experiments, these exposures were: single exposures with relatively non-aggressive strains, double exposures with non aggressive strains, and exposure to aggressive strains. Mortality increased across this gradient of exposure. In a second experiment, we varied the dose of the most aggressive P. ramosa strain and found that resisting infection when a large dose was applied resulted in greater mortality than when a medium or low dose was applied. Assuming that resistance is accomplished with an immune response, and that more aggressive parasites and/or larger doses of parasites are more immunostimulatory, these data are compatible with a cost of immunity. Indeed, in terms of survival, resisting parasites can be more harmful than infection. PMID- 17922717 TI - Individual patterns of habitat and nest-site use by hosts promote transgenerational transmission of avian brood parasitism status. AB - Brood parasitic birds impose variable fitness costs upon their hosts by causing the partial or complete loss of the hosts' own brood. Growing evidence from multiple avian host-parasite taxa indicates that exposure of individual hosts to parasitism is not necessarily random and varies with habitat use, nest-site selection, age or other phenotypic attributes. For instance, nonrandom patterns of brood parasitism had similar evolutionary consequences to those of limited horizontal transmission of parasites and pathogens across space and time and altered the dynamics of both population productivity and co-evolutionary interactions of hosts and parasites. We report that brood parasitism status of hosts of brown-headed cowbirds Molothrus ater is also transmitted across generations in individually colour-banded female prothonotary warblers Protonotaria citrea. Warbler daughters were more likely to share their mothers' parasitism status when showing natal philopatry at the scale of habitat patch. Females never bred in their natal nestboxes but daughters of parasitized mothers had shorter natal dispersal distances than daughters of nonparasitized mothers. Daughters of parasitized mothers were more likely to use nestboxes that had been parasitized by cowbirds in both the previous and current years. Although difficult to document in avian systems, different propensities of vertical transmission of parasitism status within host lineages will have critical implications both for the evolution of parasite tolerance in hosts and, if found to be mediated by lineages of parasites themselves, for the difference in virulence between such extremes as the nestmate-tolerant and nestmate-eliminator strategies of different avian brood parasite species. PMID- 17922718 TI - Evidence of an interannual effect of maternal immunization on the immune response of juveniles in a long-lived colonial bird. AB - Little is known about the maternal transfer of antibodies in natural host parasite systems despite its possible evolutionary and ecological implications. In domestic animals, the maternal transfer of antibodies can enhance offspring survival via a temporary protection against parasites, but it can also interfere with the juvenile immune response to antigens. We tested the functional role of maternal antibodies in a natural population of a long-lived colonial seabird, the kittiwake (Rissa tridactyla), using a vaccine (Newcastle disease virus vaccine) to mimic parasite exposure combined with a cross-fostering design. We first investigated the role of prior maternal exposure on the interannual transmission of Ab to juveniles. We then tested the effect of these antibodies on the juvenile immune response to the same antigen. The results show that specific maternal antibodies were transferred to chicks 1 year after maternal exposure and that these antibodies were functional, i.e. they affected juvenile immunity. These results suggest that the role of maternal antibodies may depend on the timing and pattern of offspring exposure to parasites, along with the patterns of maternal exposure and the dynamics of her immune response. Overall, our approach underlines that although the transgenerational transfer of antibodies in natural populations is likely to have broad implications, the nature of these effects may vary dramatically among host-parasite systems, depending on the physiological mechanisms involved and the ecological context. PMID- 17922719 TI - What is the required minimum landscape size for dispersal studies? AB - Among small animals dispersal parameters are mainly obtained by traditional methods using population studies of marked individuals. Dispersal studies may underestimate the rate and distance of dispersal, and be biased because of aggregated habitat patches and a small study area. The probability of observing long distance dispersal events decreases with distance travelled by the organisms. In this study a new approach is presented to solve this methodological problem. An extensive mark-release-recapture programme was performed in an area of 81 km(2) in southern Sweden. To estimate the required size of the study area for adequate dispersal measures we examined the effect of study area size on dispersal distance using empirical data and a repeated subsampling procedure. In 2003 and 2004, two species of diurnal burnet moths (Zygaenidae) were studied to explore dispersal patterns. The longest confirmed dispersal distance was 5600 m and in total 100 dispersal events were found between habitat patches for the two species. The estimated dispersal distance was strongly affected by the size of the study area and the number of marked individuals. For areas less than 10 km(2) most of the dispersal events were undetected. Realistic estimates of dispersal distance require a study area of at least 50 km(2). To obtain adequate measures of dispersal, the marked population should be large, preferably over 500 recaptured individuals. This result was evident for the mean moved distance, mean dispersal distance and maximum dispersal distance. In general, traditional dispersal studies are performed in small study areas and based on few individuals and should therefore be interpreted with care. Adequate dispersal measures for insects obtained by radio-tracking and genetic estimates (gene flow) is still a challenge for the future. PMID- 17922720 TI - Demographic response of snake-necked turtles correlates with indigenous harvest and feral pig predation in tropical northern Australia. AB - Species that mature late, experience high levels of survival and have long generation times are more vulnerable to chronic increases in mortality than species with higher fecundity and more rapid turnover of generations. Many chelonians have low hatchling survival, slow growth, delayed sexual maturity and high subadult and adult survival. This constrains their ability to respond quickly to increases in adult mortality from harvesting or habitat alteration. In contrast, the northern snake-necked turtle Chelodina rugosa (Ogilby 1890) is fast growing, early maturing and highly fecund relative to other turtles, and may be resilient to increased mortality. Here we provide correlative evidence spanning six study sites and three field seasons, indicating that C. rugosa is able to compensate demographically to conditions of relatively low subadult and adult survival, caused by pig Sus scrofa (Linnaeus 1758) predation and customary harvesting by humans. Recruitment and age specific fecundity tended to be greater in sites with low adult and subadult survival (and thus reduced densities of large turtles), owing to higher juvenile survival, a smaller size at onset of maturity and faster post-maturity growth. These patterns are consistent with compensatory density-dependent responses, and as such challenge the generality that high subadult and adult survival is crucial for achieving long-term population stability in long-lived vertebrates such as chelonians. We posit that long-lived species with 'fast' recruitment and a capacity for a compensatory demographic response, similar to C. rugosa, may be able to persist in the face of occasional or sustained adult harvest without inevitably threatening population viability. PMID- 17922721 TI - Foraging for intermittently refuged prey: theory and field observations of a parasitoid. AB - Many insect herbivores feed in concealed locations but become accessible intermittently, creating windows of greater vulnerability to attack, and generating a proportion of the prey population that is readily accessible to foraging natural enemies. We incorporated accessible prey into an extant optimal foraging model, and found that this addition allowed opportunistic exploitation of prey that have already emerged from refugia (the leaving strategy) as a viable strategy, in addition to waiting at refugia for prey to emerge (the waiting strategy). We parameterized the model empirically for the parasitoid Macrocentrus grandii and its host, Ostrinia nubilalis, under field conditions. The model predicted that M. grandii should adopt a leaving strategy when host patch density is high (travel time between patches is short), but a waiting strategy when host patch density is low (travel time between patches is long). Field observations of M. grandii patch tenure were consistent with model predictions, indicating that M. grandii exhibited flexible behaviour based on experience within a foraging bout, and that these behavioural shifts improved foraging efficiency. Behaviour of M. grandii was responsive to heterogeneity in host emergence rates, and appeared to be driven by the relatively small proportion of the host population that became accessible at a fast rate. Therefore understanding forager responses to intermittently refuged prey may require characterization of the behaviour of a subset of the prey population, rather than the average prey individual. The model can potentially be used as a framework for comparative studies across forager taxa, to understand when foragers on intermittently accessible prey should adopt fixed waiting or leaving strategies vs. a flexible strategy that is responsive to the current environment. PMID- 17922723 TI - Demographics, injury characteristics and outcome of traumatic brain injuries in northern Sweden. AB - OBJECTIVES - To describe demographics, injury characteristics and outcome of traumatic brain injury (TBI) in northern Sweden over 10 years. MATERIAL AND METHODS - Data were retrospectively collected on those individuals (n = 332) in Norrbotten, northern Sweden, with a TBI who had been transferred for neurosurgical care from 1992 to 2001. RESULTS - A majority were older men with a mild TBI and an acute or chronic subdural hematoma following a fall. Younger individuals were fewer but had more often a severe TBI from a traffic accident. Most individuals received post-acute care and brain injury rehabilitation. A majority had a moderate or severe disability, but many were discharged back home with no major changes in their physical or social environment. CONCLUSIONS - Our data confirm the relationship between age, cause of injury, injury severity and outcome in relation to TBI and underscore the need for prevention as well as the importance of TBI as a cause of long-term disability. PMID- 17922724 TI - Age-specific prevalence of epilepsy in Oppland County, Norway. AB - OBJECTIVE - To assess the prevalence of epilepsy in Oppland County, Norway, particularly with respect to age. MATERIALS AND METHODS - Population-based and hospital-based approaches were used to identify individuals with active epilepsy born in 1925, 1940, 1955, 1960 and 1970. Cases with active epilepsy were defined as individuals who had experienced more than one epileptic seizure during the last 5 years and/or were taking antiepileptic drugs. RESULTS - From a population of 11,089 persons, 90 individuals were identified with active epilepsy. The age distributed prevalence (cases per 1000 individuals born in a specified year) of active epilepsy was 10.2 for those born in 1925, 11.1 for those born in 1940, 4.5 for those born in 1955, 7.5 for those born in 1960 and 9.4 for those born in 1970. Mean epilepsy prevalence was estimated to be 8.2 cases/1000 inhabitants. CONCLUSIONS - An epilepsy prevalence of 8.2 cases/1000 inhabitants is in accordance with the results of similar studies from the western world. However, the results of such studies will vary according to the definition of 'active epilepsy'. By using the narrower International League Against Epilepsy's definition, the prevalence in our study is reduced to 5.3 cases/1000 inhabitants. PMID- 17922725 TI - Treatment of idiopathic intracranial hypertension: topiramate vs acetazolamide, an open-label study. AB - OBJECTIVES - To assess the efficacy of topiramate in the treatment of idiopathic intracranial hypertension (IIH) and to compare it with acetazolamide. METHODS - Fourty patients diagnosed as IIH and randomly assigned to treatment with either acetazolamide or topiramate were assessed prospectively. Improvement in the visual fields at the end of third, sixth and twelfth months were taken into consideration. RESULTS - The demographic, clinical features and the cerebrospinal fluid (CSF) pressure of the two treatment groups were similar at the beginning of the study. When the follow-up visual field grades were compared with the visual field grades at the beginning of the study in each group a statistically significant improvement was detected with both drugs. When the results of the two treatment groups were compared with each other no statistically significant difference was present. Prominent weight loss was recorded in the topiramate group. CONCLUSIONS - Topiramate seems to be effective in the treatment of IIH. Weight reduction as well as the reduction of the CSF formation is the possible mechanism of action. PMID- 17922726 TI - Caudate structural abnormalities in idiopathic normal pressure hydrocephalus. AB - BACKGROUND - Idiopathic normal pressure hydrocephalus (iNPH) is a reversible dementia in which fronto-striatal cognitive deficits and apathy may be present. OBJECTIVES - The study investigated structural volumetric changes in iNPH, apart from ventriculomegaly. MATERIALS AND METHODS - A full-brain voxel-based morphometric analysis between 11 iNPH patients and 14 healthy controls identified regions of interest (ROIs) for manual volumetric analyses. RESULTS - Caudate and corpus callosum ROI measurements revealed diminished caudate nuclei volume in the iNPH group. CONCLUSIONS - The role of the caudate nucleus in cognitive and affective changes in iNPH should now be explored. PMID- 17922727 TI - CSF biomarkers in the evaluation of idiopathic normal pressure hydrocephalus. AB - BACKGROUND - To evaluate cerebrospinal fluid (CSF) markers for neuronal degeneration and demyelination in idiopathic normal pressure hydrocephalus (INPH), subcortical arteriosclerotic encephalopathy (SAE), and neurologically healthy subjects. METHODS - Lumbar CSF concentrations of sulfatide, neurofilament protein light (NFL), total-tau (T-tau), hyperphosphorylated tau (P-tau), and beta amyloid(1-42) (Abeta42) were analyzed in 62 INPH patients, 26 SAE patients, and 23 neurologically healthy controls. In INPH patients, samples before and after shunt surgery were analysed. RESULTS - The CSF concentration of NFL was elevated in INPH and SAE compared with the controls, and levels of T-tau, P-tau, and Abeta42 were lower in INPH compared with SAE and controls. No difference was seen for sulfatide. All markers except Abeta42 were significantly elevated after shunt surgery. CONCLUSIONS - The most striking finding was the power of the combined pattern of NFL, P-tau, and Abeta42 in distinguishing between the clinical diagnoses of INPH, SAE, and neurologically healthy elderly. PMID- 17922728 TI - Constant blood flow reduction in premotor frontal lobe regions in ALS with dementia - a SPECT study with 3D-SSP. AB - OBJECTIVES - We investigated the regional cerebral blood flow in amyotrophic lateral sclerosis with dementia (ALS-D) patients, using single photon emission computed tomography (SPECT). MATERIALS AND METHODS - The (123)I-IMP SPECT data for 5 ALS-D and 16 ALS patients were analyzed using three-dimensional stereotactic surface projection (3D-SSP). RESULTS - 3D-SSP demonstrated marked prefrontal hypoperfusion in all the five ALS-D cases and significant bilateral prefrontal hypoperfusion in group comparisons. CONCLUSIONS - This study revealed prefrontal hypoperfusion in ALS-D cases to be an obvious abnormality with scientific objectivity. PMID- 17922729 TI - Lamotrigine in idiopathic epilepsy - increased risk of cardiac death. PMID- 17922730 TI - Sleep apnea and dialysis therapies: things that go bump in the night? AB - Sleep apnea has been linked to excessive daytime sleepiness, depressed mood, hypertension, and cardiovascular disease in the general population. The prevalence of severe sleep apnea in the conventional thrice-weekly hemodialysis population has been estimated to be more than 50%. Sleep apnea leads to repetitive episodes of hypoxemia, hypercapnia, sleep disruption, and activation of the sympathetic nervous system. The hypoxemia, arousals, and intrathoracic pressure changes associated with sleep apnea lead to sympathetic activation, endothelial dysfunction, oxidative stress, and inflammation. Because sleep apnea has been shown to be widespread in the conventional dialysis population, it may be that sleep apnea contributes substantially to the sleepiness, poor quality of life, and cardiovascular disease found in this population. The causal links between conventional dialysis and sleep apnea remain speculative, but there are likely multiple factors related to volume status and azotemia that contribute to the high rate of severe sleep apnea in dialysis patients. Both nocturnal automated peritoneal dialysis and nocturnal hemodialysis have been associated with reduced severity of sleep apnea. Nocturnal dialysis modalities may provide tools to increase our understanding of the uremic sleep apnea and may also provide therapeutic alternatives for end-stage renal disease patients with severe sleep apnea. In conclusion, sleep apnea is an important, but overlooked, public health problem for the dialysis population. The impact of sleep apnea treatment in this high-risk population may include reduced sleepiness, better mood and blood pressure, and lowered risk of cardiovascular disease. PMID- 17922731 TI - Infective endocarditis in a hemodialysis patient: a dreaded complication. AB - Infection is the most common cause of death in hemodialysis patients, after cardiovascular disease. Dialysis access infections, with secondary septicemia, contribute significantly to patient mortality. The most common source is temporary catheterization. Bacteremia occurs commonly in patients receiving hemodialysis, with infective endocarditis being a relatively uncommon, but potentially lethal complication. Valvular calcification is the most significant risk factor. The diagnosis of infective endocarditis is made clinically and confirmed with the echocardiographic modified Duke's criteria. The most common pathogen is Staphylococcus aureus and the mitral valve is the most common site. Staphylococcus aureus infective endocarditis is commonly associated with embolic phenomenon. A high index of suspicion is critical in the early recognition and management of infective endocarditis. However, prevention of bacteremia is undoubtedly the best strategy with the early placement of arteriovenous fistulae. In the case of temporary catheterization, the use of topical mupirocin or polysporin and gentamicin and/or citrate locking is beneficial. Although catheter salvage has not been studied in randomized trials, catheter removal remains standard therapy during bacteremia. PMID- 17922732 TI - A rare complication of hemodialysis catheters: superior vena cava syndrome. AB - Central venous catheters in hemodialysis patients may result in superior vena cava (SVC) syndrome. With the increasing use of these catheters, the SVC syndrome will probably be more common among hemodialysis patients. This report describes 3 cases of SVC syndrome due to central venous catheters that developed in hemodialysis patients with previous multiple catheter placements. PMID- 17922733 TI - Nonconvulsive status epilepticus: high incidence in dialysis-dependent patients. AB - Nonconvulsive status epilepticus is defined as a mental status change from baseline of at least a 30-min duration associated with constant or near continuous ictal discharges on an electroencephalogram (EEG). Nonconvulsive status epilepticus is an often-overlooked diagnosis as a cause of acute confusional state. There are few case reports in dialysis patients. We are reporting 5 patients on chronic dialysis treatment who presented with an acute confusional state. Nonconvulsive status epilepticus was diagnosed by EEG and response to intravenous benzodiazepines. We recommend early EEG to be considered in the evaluation of dialysis patients with an acute confusional state. PMID- 17922734 TI - Ectopic mediastinal parathyroid carcinoma as a cause of dialysis-dependent renal failure. AB - Carcinoma of the parathyroid gland is infrequent in patients with secondary hyperparathyroidism. Typical clinical symptoms are related to the presence of a neck mass and hypercalcemia. We describe a case of a 55-year-old man in whom primary parathyroid carcinoma led most likely to the development of end-stage dialysis-dependent renal failure, and the diagnosis of the cancer was delayed due to ectopic localization of the tumor, and dramatic complications in the clinical course of the disease, including acute pancreatitis and peritionitis. However, 6 months after successful surgery and subsequent radiotherapy, the patient is well and free from recurrences but remains chronically dialysis-dependent. PMID- 17922735 TI - Secondary sternal Aspergillus osteomyelitis in a diabetic hemodialysis patient with previous allograft rejection. AB - A 29-year-old diabetic woman who had a previously failed renal allograft on maintenance hemodialysis developed sternal aspergillosis with Aspergillus terreus following a pericardiectomy. She was successfully treated with surgical debridement and a combination of antifungal agents including amphotericin B, caspofungin, and voriconizole. The diagnostic difficulties and management are discussed. PMID- 17922736 TI - Nodular pulmonary amyloidosis and Sjogren's syndrome in a patient treated with intermittent hemodialysis. AB - In the available literature, we have found the descriptions of 5 cases of nodular pulmonary amyloidosis associated with Sjogren's syndrome. In our practice, such a case has occurred in a patient with chronic renal failure. A 53-year-old woman underwent nephrological, rheumatological, and pulmonological examinations because of end-stage renal disease with a small cirrhotic kidney in renal ultrasound examination, pulmonary nodules, and xerophthalmia. Serological data revealed a slightly positive rheumatoid factor, antinuclear antibodies, anti-SS-A, anti-SS B, and anti-RNP/Sm antibodies. Schirmer's test was positive on both sides and Sjogren's syndrome was recognized. Pulmonological examinations (the chest radiograph and CT scan, bronchofiberoscopy, culture of bronchial washings, bronchial biopsy, pleural effusion analysis, and a thick-needle biopsy) failed to determine the etiology of nodular changes in lungs. Immunofluorescence studies in the skin biopsy specimen showed IgM-positive staining. After 2 years of treatment with IHD, a toracoscopy was performed with enucleation of the nodules from the right lung. Histological examination showed massive deposits of amyloid, which allowed for a diagnosis of diffusive nodular pulmonary amyloidosis. PMID- 17922737 TI - Comparison of calcium phosphate product values using measurement of plasma total calcium and serum ionized calcium. AB - Calcium phosphate product (Ca x Pi) is a clinically relevant tool to estimate the cardiovascular risk of patients with renal failure. In reports, mostly total serum calcium has been used. As measurement of serum ionized calcium has some benefits and is being used increasingly, we estimated the respective levels of calcium phosphate product using both total (t-Ca x Pi) and ionized calcium (ion Ca x Pi). Fifty-eight healthy individuals and 180 hemodialysis (HD) patients from 2 centers were studied. Diagnostic accuracies for corresponding values of the t Ca x Pi and ion-Ca x Pi were calculated using a GraphROC program. Of HD patients, 64% had t-Ca x Pi <4.4 mmol(2)/L(2) regarded as a desirable goal, and 10% had values over 5.6 mmol(2)/L(2) associated with a high cardiovascular risk. Based on GraphROC analysis, t-Ca x Pi of 4.4 mmol(2)/L(2) corresponded to a value of 2.2 mmol(2)/L(2) of ion-Ca x Pi and, respectively, t-Ca x Pi of 5.6 mmol(2)/L(2) corresponded 2.8 mmol(2)/L(2) of ion-Ca x Pi. Owing to the good agreement between the results in the 2 centers, these values for risk levels can be used in both centers. When measurement of ionized calcium is used, Ca x Pi values of 2.2 and 2.8 mmol(2)/L(2) can be used instead of generally used values of 4.4 and 5.6 mmol(2)/L(2) with total calcium. PMID- 17922739 TI - A study of the extracorporeal rate of blood flow and blood pressure during hemodialysis. AB - Hemodynamic instability is a common problem during hemodialysis (HD). The effect of blood flow rate (BFR) on blood pressure (BP) during HD has not been previously evaluated. Subjects receiving HD for the treatment of renal failure were enrolled (n=34). For each patient, during the last hour of 2 consecutive HD sessions the BFR was set at 200 mL/min for 30 min and at 400 mL/min for 30 min, during which period the fluid removal rate was kept constant. The order of the BFR alterations was randomized. The study procedure was repeated during the next HD session but with reversal of the order of the altered BFR. During each 30-min period, BP was recorded at baseline and subsequently every 10 min. During the BFR of 400 mL/min, subjects had a higher systolic BP by an average of 4.1 mmHg compared with the BFR of 200 mL/min (95% confidence interval [CI] 0.22-7.98; p=0.038). Similarly, during the BFR of 400 mL/min, subjects had a higher diastolic BP by an average of 3.04 mmHg compared with the BFR of 200 mL/min (95% CI 0.55-5.53; p=0.017). Likewise, during the BFR of 400 mL/min, subjects had a higher mean arterial pressure by an average of 3.44 mmHg (95% CI 0.77-6.11; p=0.012). The findings suggest that during HD, BPs are maintained higher at higher BFRs as compared with lower BFRs. PMID- 17922738 TI - Effect of pharmacological suppression of secondary hyperparathyroidism on cardiovascular hemodynamics in predialysis CKD patients: A preliminary observation. AB - Cardiovascular events are the principal cause of mortality in patients with chronic kidney disease (CKD). Secondary hyperparathyroidism (SHPT), a common complication of CKD, contributes to cardiac dysfunction. This study is an attempt to demonstrate the effects of parathyroid hormone suppression with oral calcitriol on cardiovascular hemodynamics. Twenty predialysis CKD patients with SHPT were given calcitriol therapy for 12 weeks. Ten similar patients received placebo. Echocardiographic assessment of cardiac function was performed at baseline and after 12 weeks of treatment. Calcitriol therapy effectively suppressed SHPT. Baseline left ventricular (LV) end diastolic diameter and LV end systolic diameter were 4.86+/-0.48 and 2.86+/-0.33 cm, and the mean FS was 41.02+/-4.79%. Left ventricular end systolic and end diastolic volumes were normal (42.30+/-9.07 and 91.40+/-19.68 mL). The ejection fraction was slightly reduced (53.54+/-3.57%). Pretreatment Doppler indices including E velocity (0.816+/-0.087 m/s), A velocity (0.696+/-0.089 m/s), and E/A ratio (1.193+/ 0.210) were significantly impaired. After 12 weeks of calcitriol therapy, there was no significant change in the LV dimensions or ejection fraction, but there was a significant improvement in the diastolic parameters, namely the A velocity (0.680+/-0.084) and E/A ratio (1.238+/-0.180). Secondary hyperparathyroidism is an important factor in the pathogenesis of cardiovascular complications in CKD. There is evidence to support that correction of hyperparathyroidism can improve the systolic dysfunction seen in advanced kidney disease. This study shows that diastolic dysfunction seen in predialysis CKD patients may also be possibly improved with calcitriol therapy. PMID- 17922740 TI - Inadvertent postdialysis anticoagulation due to heparin line locks. AB - Large-bore dual lumen in-dwelling venous catheters are used in hemodialysis. These catheters are usually locked with heparin after the treatment. This study addressed the underappreciated postdialysis coagulopathy that can result. Thirty six patients were included: 7 dialyzed through arterio-venous fistulae, 29 through in-dwelling venous catheters. The latter group was further subdivided according to whether they received heparin or heparin-free dialysis. To assess the heparin lock, a full-dose heparin lock as well as a much weaker heparin lock and a citrate lock were used. To assess the coagulopathy, blood was taken 1 hr after dialysis. The activated partial thromboplastin time (APTT) and anti-Xa level was measured. Additionally, 6 venous catheters were removed and the amount of fluid expelled upon locking with saline was measured. Clotting from the patient group with arterio-venous fistulae was normal following dialysis. The patients with in-dwelling venous catheters and heparin locks had significantly deranged clotting; 6 out of 10 patients had abnormal APTT results. All patients with catheters, heparin-free dialysis, and heparin locks had deranged clotting (7 out of 7). The rate decreased significantly when heparinized saline was used as a lock. A subset of patients had a citrate lock rather than a heparin lock; the clotting results normalized in all but one patient. An in vitro study demonstrated immediate leakage of fluid from the end of the ports upon locking. Significant postdialysis anticoagulation can occur after dialysis, which can be attributed to the heparin line locks. This risk is considerably reduced when a citrate lock is used instead. PMID- 17922742 TI - Perihospitalization hemoglobin-epoetin associations in U.S. hemodialysis patients, 1998 to 2003. AB - While hospitalization is common for hemodialysis patients, perihospitalization associations between hemoglobin levels and epoetin doses are not well characterized. U.S. Medicare claims were used to identify 71,360 hemodialysis patients hospitalized from 1998 to 2003. Hemoglobin levels, epoetin doses, and epoetin responsiveness index (ERI) were compared by calendar year. In the prehospitalization month, the mean hemoglobin levels increased from 10.96 g/dL in 1998 to 11.76 in 2003 and the mean epoetin doses from 63,715 to 75,012 U; corresponding values in the hospitalization month were 10.53 and 11.19 g/dL, and 66,623 and 80,569 U. In each year, prehospitalization hemoglobin levels were achieved within 2 months, but ERI declined to prehospitalization levels within 12 months only in 2000. With mixed models, hemoglobin declines in the 3 prehospitalization months grew between 1998 (-0.1362 g/dL/month) and 2003 ( 0.2003 g/dL/month). Epoetin responsiveness index slopes were J-shaped, with values of 287.9, 221.1, and 356.5U/month per g/dL in 1998, 2000, and 2003. In the 3 postadmission months, a modest increase in the rapidity of hemoglobin recovery was seen (+0.2538 g/dL/month in 1998, +0.2743 in 2003), with increasing rates of ERI change (+8.7 U/month/g/dL in 1998, +146.8 in 2003). While time to recovery of prehospitalization hemoglobin levels remained constant year to year, epoetin doses and ERI did not, suggesting that optimum perihospitalization anemia management practices have yet to be determined. PMID- 17922741 TI - Sleep apnea in hemodialysis patients: risk factors and effect on survival. AB - Sleep disorders are common in patients with end-stage renal disease (ESRD). Using a simple questionnaire, we estimate the probability of sleep apnea in ESRD patients, determine the factors associated with a higher probability of sleep apnea, and determine the association between the probability of sleep apnea and cardiovascular and all-cause mortality. STUDY DESIGN: Prospective cohort study. SETTING AND PARTICIPANTS: prevalent hemodialysis patients (n=270) in 7 urban outpatient hemodialysis units. PREDICTOR: Probability of sleep apnea as quantified by the Flemons questionnaire. OUTCOMES AND MEASUREMENTS: Clinical, demographic, and dialysis-related characteristics were obtained at baseline. Total and cardiovascular mortality was ascertained after a median follow-up of 34 months. The probability of sleep apnea was low in 79 (29%) patients, moderate in 116 (43%) patients, and high in 75 (28%) patients. Male gender (odds ratio [OR] 5.13, p<0.001), obesity (BMI >30, OR 7.58, p<0.01), and interdialytic weight gain (OR 1.72/kg change, p<0.004) were independently associated with a high probability of sleep apnea. A high probability of sleep apnea at baseline did not predict total (hazard ratio [HR] 0.81, p=NS) or cardiovascular mortality (HR 0.9, p=NS). The Flemons questionnaire is validated in the general population, but has not been tested specifically in hemodialysis patients. The study may not be adequately powered to detect a difference in mortality. A high proportion of hemodialysis patients are likely to have sleep apnea; a simple bedside questionnaire can be used for screening to identify these patients. Excessive interdialytic weight gain is a potentially modifiable factor that increases the likelihood of sleep apnea. Despite the presence of a strong association between sleep apnea and mortality in the general population, a similar association could not be demonstrated in ESRD patients with a high prevalence of this condition. PMID- 17922743 TI - Relative blood volume measurements during hemodialysis: comparisons between three noninvasive devices. AB - The monitoring of relative blood volume changes (DeltaRBV) has been advocated for the prevention of hemodialysis (HD) hypotension. Stand-alone devices (Crit-Line) or devices incorporated into the HD apparatus (blood volume monitor [BVM], Hemoscan) are widely used for this purpose. Comparisons between devices are scarce. The aim of this study was, first, to compare DeltaRBV results from these 3 devices with DeltaRBV calculated from changes in laboratory-derived hemoglobin (DeltaRBV-lab-Hb) and, second, to compare DeltaRBV results between the different devices. Fourteen patients received 2 HD treatments in a randomized order: one with the Hemoscan and Crit-Line combination and one with the BVM and Crit-Line combination. DeltaRBV-lab-Hb was measured at 2 and 4 hr into the HD session. Bland-Altman analyses showed that DeltaRBV results from the 3 devices differed systematically from DeltaRBV-lab-Hb, i.e., the difference between the 3 devices and DeltaRBV-lab-Hb varied significantly (p<0.05) with the magnitude of the measurement. The interdevice comparison showed considerable differences in DeltaRBV results. At the end of the treatment, a significant difference (p<0.05) between DeltaRBV measured by the Hemoscan and Crit-Line device (-9.8+/-2.7% and 11.5+/-4%, respectively) was found. In most patients, a systematic difference between Crit-Line and Hemoscan and between Crit-Line and BVM was observed. Relative blood volume change measurements by Crit-Line, Hemoscan, and BVM yield results that differ systematically from the results obtained from laboratory derived Hb changes. Furthermore, there are substantial differences in DeltaRBV results between the 3 DeltaRBV devices. PMID- 17922744 TI - Aging veterans and the end-stage renal disease management dilemma in the millennium. AB - The population of aging veterans with complex multiple medical problems is increasing steadily in developed nations. The life expectancy in an aging population with end-stage renal disease (ESRD) is often compared with terminal malignancy. Renal failure in elderly patients often generates a myriad of complicated issues and the nephrologists are faced with the dilemma of conveying the prognosis of renal failure in elderly patients and also explain the pros and cons of offering a renal replacement therapy. Our objectives were to assess the cumulative survival in veterans with ESRD over 70 years of age and to evaluate the factors considered for either not initiating or withdrawing from dialysis. All veterans above age 70 years, who were being evaluated for possible dialysis therapy over a 5-year period, were included in the study. The cumulative survival rates at 1 year, 3 years and 5 years were 60%, 37%, and 20%, respectively. Tunneled cuffed catheter was the dialysis access in a third of these patients on dialysis adding to the morbidity. Twenty-four patients considered either not initiating or withdrawing from dialysis therapy after consensus agreement from either the patient or the power of attorney. The decision to initiate dialysis therapy should be made considering the social, ethical, and associated comorbid conditions. A decision to not initiate or withdraw dialysis is possible in critically ill elderly patients and if taken judiciously can reduce physical and mental stress of both the patient and their family members. PMID- 17922745 TI - Does a history of peritoneal dialysis result in an impaired gastrointestinal life quality? AB - Peritoneal dialysis (PD) invariably induces sclerotic changes in the peritoneal membrane. The impact of these changes on the well-being of PD patients has not been studied sufficiently. In a matched-pair analysis, the gastrointestinal life quality of patients with a history of PD was compared with end-stage renal disease patients who never performed PD, using a standardized questionnaire (gastrointestinal life-quality index [GLQI]). We identified all patients in our dialysis unit who underwent PD between 1989 and 2001 and who were alive in October 2001 (PD patients; n=53). Patients who were treated by hemodialysis (HD patients) were recruited as pairs. Hemodialysis and PD patients did not differ in gastrointestinal life quality (GLQI: HD 106.0+/-16.4 points; PD 104.0+/-16.7 points; p=0.70). Gastrointestinal life quality was neither correlated with the number of peritonitis episodes, nor with the duration of PD treatment. Peritoneal dialysis treatment is not associated with a long-term impairment of gastrointestinal life quality. PMID- 17922747 TI - What is a surrogate marker for optimal dialysis? AB - To find a surrogate marker to obtain optimal dialysis delivery from the viewpoint of nutrition, 180 maintenance hemodialysis patients (109 males/71 females) were enrolled between October 1999 and June 2006 at our kidney center. In the 449 hemodialysis treatments, ultrapure dialysis solutions and high-flux synthetic membranes were utilized. Parameters were measured by Kt/V(urea) and postdialysis urea rebound, Kc (the cellular membrane clearance for urea), urea clear space (CS), %creatinine generation rate, %lean body mass, total body water, and so on. We examined the correlation between dialysis delivery and nutritional parameters: Kt/V(urea) and postdialysis urea rebound were found to be strongly and negatively correlated with nutritional parameters. However, Kc and CS have shown positive and strong correlations with nutritional parameters such as %creatinine generation rate, %lean body mass, and total body water as well. In addition, the age factor was correlated with Kt/V(urea) positively, and it influenced Kc and CS negatively. As a conventional dialysis parameter, Kt/V(urea) did not reflect nutrition, but Kc was found to improve nutrition due to the increase of the dialysis delivery. Therefore, Kc might be a reliable surrogate marker for optimal dialysis. PMID- 17922746 TI - A comparison of center-based vs. home-based daily hemodialysis for patients with end-stage renal disease. AB - Home hemodialysis has been a therapeutic option for almost 4 decades. The complexity of dialysis equipment has been a factor-limiting adoption of this modality. We performed a feasibility study to demonstrate the safety of center based vs. home-based daily hemodialysis with the NxStage System One portable hemodialysis device. We also performed a retrospective analysis to determine if clinical effects previously associated with short-daily dialysis were also seen using this novel device. We conducted a prospective, 2-treatment, 2-period, open label, crossover study of in-center hemodialysis vs. home hemodialysis in 32 patients treated at 6 U.S. centers. The 8-week In-Center Phase (6 days/week) was followed by a 2-week transition period and then followed by the 8-week Home Phase (6 days/week). We retrospectively collected data on hemodialysis treatment parameters immediately preceding the study in a subset of patients. Twenty-six out of 32 patients (81%) successfully completed the study. Successful delivery of at least 90% of prescribed fluid volume (primary endpoint) was achieved in 98.5% of treatments in-center and 97.3% at home. Total effluent volume as a percentage of prescribed volume was between 94% and 100% for all study weeks. The composite rate of intradialytic and interdialytic adverse events per 100 treatments was significantly higher for the In-Center Phase (5.30) compared with the Home Phase (2.10; p=0.007). Compared with the period immediately preceding the study, there were reductions in blood pressure, antihypertensive medications, and interdialytic weight gain. Daily home hemodialysis with a small, easy-to-use hemodialysis device is a viable dialysis option for end-stage renal disease patients capable of self/partner-administered dialysis. PMID- 17922748 TI - Challenges and limitations of maintenance hemodialysis in urban South India. AB - Maintenance hemodialysis is a treatment modality available to few patients reaching end-stage renal disease in India. However, the morbidity and outcome of such treatment remains largely unknown. A retrospective cohort of patients commencing hemodialysis in a secondary care institution in India between January 1, 2002 and December 31, 2004 was studied. Patient demographics, cardiac status, access, hospitalizations, and emergency room visits were assessed and outcomes determined. During the study period, 95 patients (66 males, 29 females) commenced maintenance hemodialysis. The underlying cause of chronic kidney disease was diabetic nephropathy in 66.3% of patients. Cumulative follow-up was 676+9.1 patient months. The mean serum creatinine (+SD) at initiation of dialysis was 8.39+3.28 mg%. Thirty-six percent of patients had a functioning arteriovenous fistula at commencement of dialysis, while the remaining 64% of patients required temporary access. The mean number of comorbidities was 1.9+1.0/patient; diastolic dysfunction was deemed to be present in 20.4% of the patients. The hospitalization rate was 3.9/patient year; the number of visits to the emergency room was 4.9/patient year. Cardiac pathology was the most common cause leading to hospitalization and emergency room visits. Diabetic patients were older and had higher cardiac morbidity (p<0.01). The outcome was as follows: 39% transferred to other units; 27% died; 9% switched to CAPD; 8% lost to follow-up; 1% transplantation: Kaplan-Meier survival analysis showed a median survival of 410 days. Patients commencing hemodialysis in an urban dialysis center in South India are predominantly male and have significant comorbidity including diabetes and cardiac disease. Outcome is generally poor. Hence, a huge opportunity for improvement exists. PMID- 17922749 TI - Effect of intravenous iron on insulin sensitivity in dialysis patients. PMID- 17922750 TI - Steering directed protein evolution: strategies to manage combinatorial complexity of mutant libraries. AB - How to explore protein sequence space efficiently and how to generate high quality mutant libraries that allow to identify improved variants with current screening technologies are key questions for any directed protein evolution experiment. High-quality mutant libraries can be generated through improved random mutagenesis methodologies and by restricting diversity generation through computational methods to residues which have high success probabilities. Advances in mutant library design and computational tools to focus diversity generation are summarized in this minireview and discussed from an experimentalist point of view in the context of directed protein evolution. PMID- 17922751 TI - Crash of a population of the marine heterotrophic flagellate Cafeteria roenbergensis by viral infection. AB - Viruses are known as important mortality agents of marine microorganisms. Most studies focus on bacterial and algal viruses, and few reports exist on viruses infecting marine heterotrophic protists. Here we show results from several incubations initiated with a microbial assemblage from the central Indian Ocean and amended with different amounts of organic matter. Heterotrophic flagellates developed up to 30,000 cells ml(-1) in the most enriched incubation. A 18S rDNA clone library and fluorescent in situ hybridization counts with newly designed probes indicated that the peak was formed by Cafeteria roenbergensis and Caecitellus paraparvulus (90% and 10% of the cells respectively). Both taxa were below detection in the original sample, indicating a strong positive selective bias during the enrichment. During the peak, C. roenbergensis cells were observed with virus-like particles in the cytoplasm, and 4 days later this taxa could not be detected. Transmission electron microscopy confirmed the viral nature of these particles, which were large (280 nm), had double-stranded DNA, and were produced with a burst size of approximately 70. This virus was specific of C. roenbergensis as neither C. paraparvulus that was never seen infected, nor other flagellate taxa that developed in later stages of the incubation, appeared attacked. This is one of the few reports on a heterotrophic flagellate virus and the implications of this finding in the Indian Ocean are discussed. PMID- 17922752 TI - Patterns of bacterial diversity across a range of Antarctic terrestrial habitats. AB - Although soil-borne bacteria represent the world's greatest source of biological diversity, it is not well understood whether extreme environmental conditions, such as those found in Antarctic habitats, result in reduced soil-borne microbial diversity. To address this issue, patterns of bacterial diversity were studied in soils sampled along a > 3200 km southern polar transect spanning a gradient of increased climate severity over 27 degrees of latitude. Vegetated and fell-field plots were sampled at the Falkland (51 degrees S), South Georgia (54 degrees S), Signy (60 degrees S) and Anchorage Islands (67 degrees S), while bare frost sorted soil polygons were examined at Fossil Bluff (71 degrees S), Mars Oasis (72 degrees S), Coal Nunatak (72 degrees S) and the Ellsworth Mountains (78 degrees S). Bacterial 16S rRNA gene sequences were recovered subsequent to direct DNA extraction from soil, polymerase chain reaction amplification and cloning. Although bacterial diversity was observed to decline with increased latitude, habitat-specific patterns appeared to also be important. Namely, a negative relationship was found between bacterial diversity and latitude for fell-field soils, but no such pattern was observed for vegetated sites. The Mars Oasis site, previously identified as a biodiversity hotspot within this region, proved exceptional within the study transect, with unusually high bacterial diversity. In independent analyses, geographical distance and vegetation cover were found to significantly influence bacterial community composition. These results provide insight into the factors shaping the composition of bacterial communities in Antarctic terrestrial habitats and support the notion that bacterial diversity declines with increased climatic severity. PMID- 17922753 TI - A dual biosensor for 2-alkyl-4-quinolone quorum-sensing signal molecules. AB - Pseudomonas, Burkholderia and Alteromonas species produce diverse 2-alkyl-4 quinolones (AHQs) which inhibit the growth of bacteria, algae and phytoplankton, chelate iron, modulate mammalian host immune defences and act as quorum-sensing (QS) signal molecules. To facilitate the detection, identification and quantification of the major Pseudomonas aeruginosa AHQs 2-heptyl-3-hydroxy-4 quinolone (PQS) and 2-heptyl-4-quinolone (HHQ) we developed two different AHQ biosensors. These were constructed by introducing either a lecA::luxCDABE or a pqsA::luxCDABE reporter gene fusion into a P. aeruginosa pqsA mutant which cannot synthesize AHQs. While both biosensors responded similarly to PQS (EC(50) 18 +/- 4 microM), the pqsA::luxCDABE biosensor was most sensitively activated by HHQ (EC(50) 0.44 +/- 0.1 microM). This biosensor was also activated albeit less sensitively by (i) PQS analogues with alkyl chains varying from C1 to C11, (ii) HHQ analogues with C9 and C11 alkyl chains and (iii) 2-heptyl-4-hydroxyquinoline N-oxide (HHQNO). The AHQ biosensor also responded differentially to the AHQs present in cell free culture supernatants prepared from PAO1 and isogenic strains carrying mutations in genes (pqsA, pqsH, lasR, lasI, rhlR, rhlI) known to influence AHQ production. The AHQ profiles of P. aeruginosa strains was also evaluated by overlaying thin layer chromatogram (TLC) plates with the pqsA::luxCDABE biosensor. In PAO1, three major bioluminescent spots were observed which correspond to PQS, HHQ and a mixture of 2 nonyl-4-quinolone and HHQNO. We also noted that on TLC plates the biosensor not only produced bioluminescence in response to AHQs but also the green pigment, pyocyanin which offers an alternative visual indicator for AHQ production. PMID- 17922754 TI - Elucidation of metabolic pathways in glycogen-accumulating organisms with in vivo 13C nuclear magnetic resonance. AB - Glycogen-accumulating organisms (GAOs) are found in enhanced biological phosphorus removal systems where they compete with polyphosphate-accumulating organisms for external carbon substrates. (13)C nuclear magnetic resonance ((13)C NMR) was used to elucidate the metabolic pathways operating in an enriched GAO culture dominated by two known GAOs (81.2%). The experiments consisted of adding (13)C-acetate (labelled on position 1 or 2) to the culture under anaerobic conditions, and operating the culture through a cycle consisting of an anaerobic, an aerobic and a further anaerobic phase. The carbon transformations over the cycle were monitored using in vivo(13)C-NMR. The two-carbon moieties in hydroxybutyrate and hydroxyvalerate were derived from acetate, while the propionyl precursor of hydroxyvalerate was primarily derived from glycogen, with only a small fraction originating from acetate. Comparison of the labelling patterns in hydroxyvalerate at the end of the first and the second anaerobic periods in pulse experiments with 2-(13)C-acetate showed that the Entner Doudoroff (ED) pathway was used for the breakdown of glycogen. This conclusion was further supported by the labelling pattern on glycogen observed in the pulse experiments with 1-(13)C-acetate, which can only be explained by the operation of ED with recycling of pyruvate and glyceraldehyde 3-phosphate via gluconeogenesis. The activity of the ED pathway for glycogen degradation by GAOs is demonstrated here for the first time. In addition, the decarboxylating part of the tricarboxylic acid cycle was confirmed to operate also under anaerobic conditions. PMID- 17922755 TI - Metagenomic analysis of the microbial community associated with the coral Porites astreoides. AB - The coral holobiont is a dynamic assemblage of the coral animal, zooxanthellae, endolithic algae and fungi, Bacteria,Archaea and viruses. Zooxanthellae and some Bacteria form relatively stable and species-specific associations with corals. Other associations are less specific; coral-associated Archaea differ from those in the water column, but the same archaeal species may be found on different coral species. It has been hypothesized that the coral animal can adapt to differing ecological niches by 'switching' its microbial associates. In the case of corals and zooxanthellae, this has been termed adaptive bleaching and it has important implications for carbon cycling within the coral holobiont and ultimately the survival of coral reefs. However, the roles of other components of the coral holobiont are essentially unknown. To better understand these other coral associates, a fractionation procedure was used to separate the microbes, mitochondria and viruses from the coral animal cells and zooxanthellae. The resulting metagenomic DNA was sequenced using pyrosequencing. Fungi, Bacteria and phage were the most commonly identified organisms in the metagenome. Three of the four fungal phyla were represented, including a wide diversity of fungal genes involved in carbon and nitrogen metabolism, suggesting that the endolithic community is more important than previously appreciated. In particular, the data suggested that endolithic fungi could be converting nitrate and nitrite to ammonia, which would enable fixed nitrogen to cycle within the coral holobiont. The most prominent bacterial groups were Proteobacteria (68%), Firmicutes (10%), Cyanobacteria (7%) and Actinobacteria (6%). Functionally, the bacterial community was primarily heterotrophic and included a number of pathways for the degradation of aromatic compounds, the most abundant being the homogentisate pathway. The most abundant phage family was the ssDNA Microphage and most of the eukaryotic viruses were most closely related to those known to infect aquatic organisms. This study provides a metabolic and taxonomic snapshot of microbes associated with the reef-building coral Porites astreoides and presents a basis for understanding how coral-microbial interactions structure the holobiont and coral reefs. PMID- 17922756 TI - Resource dynamics during infection of Micromonas pusilla by virus MpV-Sp1. AB - Viruses infecting marine phytoplankton drive phytoplankton diversity, terminate blooms and shuttle genetic material. Assessments of the scale of viral impacts on trophic networks are, however, speculative. We investigated fluxes of DNA between host and virus during infection of the prasinophyte alga Micromonas pusilla by phycodnavirus MpV SP1. Under a light-dark regimen, viral genomes accumulated to a transient peak within 24 h, at the expense of both host DNA synthesis and nuclear DNA. Viral genome abundance then declined soon after host lysis. This release of a phosphate-rich nucleotide pool during viral infection of phytoplankton should be considered in trophic models. Lysis required light and was stalled in darkness, meanwhile viral genome replication proceeded slowly in the dark. Viral exploitation of this host is therefore only partially light-dependent and infected phytoplankton are poised to lyse at dawn or if mixed to the photic zone. The chloroplast genome remained intact until lysis, indicating that either this DNA pool is inaccessible or the virus spares the chloroplast for its energy and reductant generation. The photochemical turnover of residual Photosystem II complexes accelerated during lysis, indicating that events in late infection heighten demands on the remaining host photosynthetic systems, consistent with the light dependency of lysis. PMID- 17922757 TI - Intra- and inter-lake variability of free-living and particle-associated Actinobacteria communities. AB - We have analysed the inter- and intra-lake variability of free-living and particle-associated freshwater Actinobacteria communities in four limnological different lakes of the Mecklenburg Lake District, Northeastern Germany. Denaturing gradient gel electrophoresis (DGGE) specific for Actinobacteria was used to investigate phylogenetic diversity and seasonal dynamics of actinobacterial communities in the epilimnion of all lakes (inter-lake variability) and to assess differences between Actinobacteria communities of the epi-, meta- and hypolimnion of a single lake (intra-lake variability) respectively. DGGE analyses showed significant inter- and intra-lake differences between Actinobacteria communities of all lakes and water layers as well as between free-living and particle-associated Actinobacteria. Phylogenetic inferences of 16S rRNA gene sequences suggest that particular members of particle associated Actinobacteria were exclusively affiliated to certain actinobacterial lineages. The phylogenetic comparison of 16S rRNA gene sequences of all lakes and water layer, however, indicated the occurrence of almost similar phylogenetic lineages in all studied habitats and suggest high intracluster diversity within already known actinobacterial lineages. Non-metric multidimensional scaling (NMS) ordination analyses and Pearson's product moment correlations revealed several strong correlations between the investigated Actinobacteria communities and various limnological parameters, such as conductivity, total phosphorous, alkalinity or primary production. However, no uniform correlation patterns were found between lakes, water layers and bacterial fractions. These heterogeneous correlation patterns together with the phylogenetic similarities of Actinobacteria communities from different lakes indicate that particular Actinobacteria represent various ecotypes or exhibit a pronounced ecophysiological plasticity. PMID- 17922758 TI - Transcriptional response of Silicibacter pomeroyi DSS-3 to dimethylsulfoniopropionate (DMSP). AB - Dimethylsufoniopropionate (DMSP) is an abundant organic sulfur compound in the ocean and an important substrate for marine bacterioplankton. The Roseobacter clade of marine alphaproteobacteria, including Silicibacter pomeroyi strain DSS 3, are known to be involved in DMSP degradation in situ. The fate of DMSP has important implications for the global sulfur cycle, but the genes involved in this process and their regulation are largely unknown. S. pomeroyi is capable of performing two major pathways of DMSP degradation, making it an important model organism. Based on the full genome sequence of this strain we designed an oligonucleotide-based microarray for the detection of transcripts of nearly all genes. The array was used to study the transcriptional response of S. pomeroyi cultures to additions of DMSP compared to the non-sulfur compound acetate in a time series experiment. We identified a number of upregulated genes that could be assigned to potential roles in the metabolism of DMSP. DMSP also affected the transcription of genes for transport and metabolism of peptides, amino acids and polyamines. DMSP concentration may thus also play a role as a chemical signal, indicating phytoplankton abundance and eliciting a regulatory response aimed at making maximum use of available nutrients. PMID- 17922759 TI - Rare actinomycete bacteria from the shallow water sediments of the Trondheim fjord, Norway: isolation, diversity and biological activity. AB - Actinomycete bacteria produce a wide variety of secondary metabolites with diverse biological activities, some of which have been developed for human medicine. Rare actinomycetes are promising sources in search for new drugs, and their potential for producing biologically active molecules is poorly studied. In this work, we have investigated the diversity of actinomycetes in the shallow water sediments of the Trondheim fjord (Norway). Due to the use of different selective isolation methods, an unexpected variety of actinomycete genera was isolated. Although the predominant genera were clearly Streptomyces and Micromonospora, representatives of Actinocorallia, Actinomadura, Knoellia, Glycomyces, Nocardia, Nocardiopsis, Nonomuraea, Pseudonocardia, Rhodococcus and Streptosporangium genera were isolated as well. To our knowledge, this is the first report describing isolation of Knoellia and Glycomyces species from the marine environment. 35 selected actinomycete isolates were characterized by 16S rDNA sequencing, and were shown to represent strains from 11 different genera. In addition, these isolates were tested for antimicrobial activity and the presence of polyketide synthase and non-ribosomal peptide synthetase genes. This study confirms the significant biodiversity of actinobacteria in the Norwegian marine habitats, and their potential for producing biologically active compounds. PMID- 17922760 TI - Submersed macrophytes play a key role in structuring bacterioplankton community composition in the large, shallow, subtropical Taihu Lake, China. AB - Within-lake horizontal heterogeneity of bacterioplankton community composition (BCC) was investigated in the large and shallow subtropical Taihu Lake (2338 km(2), maximum depth < 3 m). Samples were collected at 17 sites along a trophic gradient ranging from mesotrophic to hypertrophic areas in August and September 2004. These sites cover two alternative stable states of shallow lakes, which are basically characterized by the dominance or the lack of submerged macrophytes. In the case of Taihu Lake, the macrophyte-dominated state is characterized by clear water and immobilized sediment, and the state largely lacking macrophytes is characterized by the dominance of phytoplankton, frequent wind-driven re suspension of sediments, and a high turbidity. Three different methods, i.e. denaturing gradient gel electrophoresis (DGGE), reverse line blot hybridization (RLB) with probes targeting 17 freshwater bacterial groups, and 16S rRNA gene cloning and sequencing, were used for analysis of BCC. The BCC varied strongly between the two alternative ecological states, but less pronounced between phytoplankton-dominated sites even spanning chlorophyll a gradients from 16.5 (mesotrophic) to 229.8 microg l(-1) (hypertrophic). The 16S rRNA gene library representing the turbid water state contained many sequences closely related to sequences previously obtained from soil or freshwater sediment samples. Furthermore, sequences representing two new lineages of freshwater Actinobacteria were obtained from the investigated samples. Comparative statistical analyses of BCC along the investigated ecological gradients revealed that the dominance of submersed macrophytes was the most influential factor on BCC, responsible for a major part of the observed within-habitat heterogeneity of BCC in Taihu Lake. PMID- 17922761 TI - Grazing protozoa and magnetosome dissolution in magnetotactic bacteria. AB - Magnetotactic bacteria show an ability to navigate along magnetic field lines because of magnetic particles called magnetosomes. All magnetotactic bacteria are unicellular except for the multicellular prokaryote (recently named 'Candidatus Magnetoglobus multicellularis'), which is formed by an orderly assemblage of 17 40 prokaryotic cells that swim as a unit. A ciliate was used in grazing experiments with the M. multicellularis to study the fate of the magnetosomes after ingestion by the protozoa. Ciliates ingested M. multicellularis, which were located in acid vacuoles as demonstrated by confocal laser scanning microscopy. Transmission electron microscopy and X-ray microanalysis of thin-sectioned ciliates showed the presence of M. multicellularis and magnetosomes inside vacuoles in different degrees of degradation. The magnetosomes are dissolved within the acidic vacuoles of the ciliate. Depending on the rate of M. multicellularis consumption by the ciliates the iron from the magnetosomes may be recycled to the environment in a more soluble form. PMID- 17922762 TI - Heterotrophic symbionts of phototrophic consortia: members of a novel diverse cluster of Betaproteobacteria characterized by a tandem rrn operon structure. AB - Phototrophic consortia represent the most highly developed type of interspecific association of bacteria and consist of green sulfur bacterial epibionts attached around a central colourless rod-shaped bacterium. Based on 16S rRNA gene sequencing, the central bacterium of the consortium 'Chlorochromatium aggregatum' was recently shown to represent a novel and phylogenetically isolated lineage of the Comamonadaceae within the beta-subgroup of the Proteobacteria. To date, 19 types of phototrophic consortia are distinguished based on the different 16S rRNA gene sequences of their epibionts, but the diversity and phylogenetic relationships of the heterotrophic partner bacteria are still unknown. We developed an approach based on the specific rrn (ribosomal RNA) operon structure of the central bacterium of 'C. aggregatum' to recover 16S rRNA gene sequences of other central bacteria and their close relatives from natural consortia populations. Genomic DNA of the central bacterium of 'C. aggregatum' was first enriched several hundred-fold by employing a selective method for growth of consortia in a monolayer biofilm followed by a purification of the genome of the central bacterium by cesium chloride-bisbenzimidazole equilibrium density gradient centrifugation. A combination of inverse PCR, cloning and sequencing revealed that two rrn operons of the central bacterium are arranged in a tandem fashion and are separated by an unusually short intergenic region of 195 base pairs. This rare gene order was exploited to screen various natural microbial communities by PCR. We discovered a diverse and previously unknown subgroup of Betaproteobacteria in the chemoclines of freshwater lakes. This group was absent in other freshwater and soil samples. All the 16S rRNA gene sequences recovered are related to that of the central bacterium of 'C. aggregatum'. Fluorescence in situ hybridization indicated that two of these sequences originated from central bacteria of different phototrophic consortia, which, however, were only distantly related to the central bacterium of 'C. aggregatum'. Based on a detailed phylogenetic analysis, these central bacterial symbionts of phototrophic consortia have a polyphyletic origin. PMID- 17922763 TI - Investigations of the structure and function of bacterial communities associated with Sphagnum mosses. AB - High acidity, low temperature and extremely low concentration of nutrients form Sphagnum bogs into extreme habitats for organisms. Little is known about the bacteria associated with living Sphagnum plantlets, especially about their function for the host. Therefore, we analysed the endo- and ectophytic bacterial populations associated with two widely distributed Sphagnum species, Sphagnum magellanicum and Sphagnum fallax, by a multiphasic approach. The screening of 1222 isolates for antagonistic activity resulted in 326 active isolates. The bacterial communities harboured a high proportion of antifungal (26%) but a low proportion of antibacterial isolates (0.4%). Members of the genus Burkholderia (38%) were found to be the most dominant group of antagonistic bacteria. The finding that a large proportion (89%) of the antagonistic bacteria produced antifungal compounds may provide an explanation for the well-known antimicrobial activity of certain Sphagnum species. The secondary metabolites of the Sphagnum species themselves were analysed by HPLC-PDA. The different spectra of detected compounds may not only explain the antifungal activity but also the species specificity of the microbial communities. The latter was analysed using cultivation-independent single-stranded conformation polymorphism (SSCP) analysis. Using Burkholderia-specific primers we found a high diversity of Burkholderia isolates in the endophytic and ectophytic habitats of Sphagnum. Furthermore, a high diversity of nitrogen-fixing bacteria was detected by using nifH-specific primers, especially inside Sphagnum mosses. In conclusion, this study provides evidence that both Sphagnum species were colonized by characteristic bacterial populations, which appear to be important for pathogen defence and nitrogen fixation. PMID- 17922764 TI - Spatial variability of sulfate reduction in a shallow aquifer. AB - The distribution and metabolic activity of sulfate-reducing bacteria (SRB) in a shallow, suboxic aquifer were studied. A radioimaging technique was used to visualize and quantify the activity of sulfate reducers in sediments at a centimetre-level scale. The distribution of SRB metabolic activity was heterogeneous with areas showing little activity far outnumbering areas with high activity. Variation in sulfate-reducing activity was not statistically correlated with variation in depth, bacterial numbers, or the following sediment properties: sediment type (sand, peat or silt), grain size, permeability and hydraulic conductivity. Sulfate-reducing bacteria activity did vary significantly with sediment porosity (multivariate analysis, r = 0.48). We hypothesized that the small pore sizes associated with sediments with low porosity restricted the ability of SRB to grow to high numbers as well as their access to nutrients. To further explore the relationship between pore size and microbial metabolic activity, columns with varying pore diameters were constructed. Sulfate-reducing bacteria in the columns with the smallest pore diameters had the lowest rates of metabolism and SRB metabolic rates increased as the pore diameter increased. For the aquifer studied, sediment porosities and pore sizes were the main factor controlling SRB activity. PMID- 17922765 TI - Exploration of intraclonal adaptation mechanisms of Pseudomonas brassicacearum facing cadmium toxicity. AB - Pseudomonas brassicacearum forms phenotypic variants in vitro as well as in planta during root colonization under natural conditions, leading to subpopulations (phase I and II cells) that differ in colony morphology and production of exoenzymes/secondary metabolites. The maximal concentration of cadmium allowing both variants growth was 25 muM; however, phase II cells accumulated fivefold higher Cd than phase I cells, even though both variants showed the same growth rate and kinetics, comprising a long stasis period (50 h). The whole transcriptome analysis of both variants in response to Cd was investigated using the home-made DNA microarrays. This analysis revealed completely different adaptation mechanisms developed by each variant to withstand and grow in the presence of the toxic. A re-organization of the cell wall to limit Cd entrance was noticed for phase I cells, as genes encoding levan exopolymers were downregulated at the expense of an upregulation of genes encoding alginate, and an upregulation of transporters such as cadA, and a downregulation of copper transporters. Phase II cells were unable to prevent Cd entrance and recruited genes under the control of oxyR and soxR regulation to face osmotic and oxidant stresses generated by Cd. Putrescine and spermidine metabolism appeared to play a central role in Cd tolerance. Microarray data were validated by biological analyses such as motility, oxidative stress assay, metabolite profiling with ICR-FT/MS and UPLC, capillary electrophoresis analysis of biogenic amines. PMID- 17922767 TI - Biofilm formation in Desulfovibrio vulgaris Hildenborough is dependent upon protein filaments. AB - Desulfovibrio vulgaris Hildenborough is a Gram-negative sulfate-reducing bacterium (SRB), and the physiology of SRBs can impact many anaerobic environments including radionuclide waste sites, oil reservoirs and metal pipelines. In an attempt to understand D. vulgaris as a population that can adhere to surfaces, D. vulgaris cultures were grown in a defined medium and analysed for carbohydrate production, motility and biofilm formation. Desulfovibrio vulgaris wild-type cells had increasing amounts of carbohydrate into stationary phase and approximately half of the carbohydrate remained internal. In comparison, a mutant that lacked the 200 kb megaplasmid, strain DeltaMP, produced less carbohydrate and the majority of carbohydrate remained internal of the cell proper. To assess the possibility of carbohydrate re allocation, biofilm formation was investigated. Wild-type cells produced approximately threefold more biofilm on glass slides compared with DeltaMP; however, wild-type biofilm did not contain significant levels of exopolysaccharide. In addition, stains specific for extracellular carbohydrate did not reveal polysaccharide material within the biofilm. Desulfovibrio vulgaris wild-type biofilms contained long filaments as observed with scanning electron microscopy (SEM), and the biofilm-deficient DeltaMP strain was also deficient in motility. Biofilms grown directly on silica oxide transmission electron microscopy (TEM) grids did not contain significant levels of an exopolysaccharide matrix when viewed with TEM and SEM, and samples stained with ammonium molybdate also showed long filaments that resembled flagella. Biofilms subjected to protease treatments were degraded, and different proteases that were added at the time of inoculation inhibited biofilm formation. The data indicated that D. vulgaris did not produce an extensive exopolysaccharide matrix, used protein filaments to form biofilm between cells and silica oxide surfaces, and the filaments appeared to be flagella. It is likely that D. vulgaris used flagella for more than a means of locomotion to a surface, but also used flagella, or modified flagella, to establish and/or maintain biofilm structure. PMID- 17922768 TI - Identification of active methanotrophs in a landfill cover soil through detection of expression of 16S rRNA and functional genes. AB - Active methanotrophs in a landfill soil were revealed by detecting the 16S rRNA of methanotrophs and the mRNA transcripts of key genes involved in methane oxidation. New 16S rRNA primers targeting type I and type II methanotrophs were designed and optimized for analysis by denaturing gradient gel electrophoresis. Direct extraction of RNA from soil enabled the analysis of the expression of the functional genes: mmoX, pmoA and mxaF, which encode subunits of soluble methane monooxygenase, particulate methane monooxygenase and methanol dehydrogenase respectively. The 16S rRNA polymerase chain reaction (PCR) primers for type I methanotrophs detected Methylomonas, Methylosarcina and Methylobacter sequences from both soil DNA and cDNA which was generated from RNA extracted directly from the landfill cover soil. The 16S rRNA primers for type II methanotrophs detected primarily Methylocella and some Methylocystis 16S rRNA genes. Phylogenetic analysis of mRNA recovered from the soil indicated that Methylobacter, Methylosarcina, Methylomonas, Methylocystis and Methylocella were actively expressing genes involved in methane and methanol oxidation. Transcripts of pmoA but not mmoX were readily detected by reverse transcription polymerase chain reaction (RT-PCR), indicating that particulate methane monooxygenase may be largely responsible for methane oxidation in situ. PMID- 17922766 TI - Aerosolization of mycobacteria and legionellae during dental treatment: low exposure despite dental unit contamination. AB - Dental unit waterlines (DUWL) support growth of a dense microbial population that includes pathogens and hypersensitivity-inducing bacteria, such as Legionella spp. and non-tuberculous mycobacteria (NTM). Dynamic dental instruments connected to DUWL generate aerosols in the work environment, which could allow waterborne pathogens to be aerosolized. The use of the real-time quantitative polymerase chain reaction (qPCR) provides a more accurate estimation of exposure levels compared with the traditional culture approach. Bioaerosol sampling was performed 13 times in an isolated dental treatment room according to a standardized protocol that included four dental prophylaxis treatments. Inhalable dust samples were taken at the breathing zone of both the hygienist and patient and outside the treatment room (control). Total bacteria as well as Legionella spp. and NTM were quantified by qPCR in bioaerosol and DUWL water samples. Dental staff and patients are exposed to bacteria generated during dental treatments (up to 4.3 E + 05 bacteria per m(3) of air). Because DUWL water studied was weakly contaminated by Legionella spp. and NTM, their aerosolization during dental treatment was not significant. As a result, infectious and sensitization risks associated with legionellae and NTM should be minimal. PMID- 17922770 TI - Hexavalent uranium supports growth of Anaeromyxobacter dehalogenans and Geobacter spp. with lower than predicted biomass yields. AB - The stimulation of bacteria capable of reducing soluble U(VI) to sparingly soluble U(IV) is a promising approach for containing U(VI) plumes. Anaeromyxobacter dehalogenans is capable of mediating this activity; however, its ability to couple U(VI) reduction to growth has not been established. Monitoring the increase in 16S rRNA gene copy numbers using quantitative real-time PCR (qPCR) in cultures provided with U(VI) as an electron acceptor demonstrated growth, and 7.7-8.6 x 10(6) cells were produced per mumole of U(VI) reduced. This biomass yield was lower than predicted based on the theoretical free energy changes associated with U(VI)-to-U(IV) reduction. Lower than predicted growth yields with U(VI) as electron acceptor were also determined in cultures of Geobacter lovleyi and Geobacter sulfurreducens suggesting that U(VI) reduction is inefficient or imposes an additional cost to growing cells. These findings have implications for U(VI) bioremediation because Anaeromyxobacter spp. and Geobacter spp. contribute to radionuclide immobilization in contaminated subsurface environments. PMID- 17922771 TI - The bacterial aetiology of rosy discoloration of ancient wall paintings. AB - The inventory of microorganisms responsible for biological deterioration of ancient paintings has become an integral part of restoration activities. Here, the microbial agent of rosy discoloration on medieval frescoes in the Crypt of the Original Sin (Matera, Italy) was investigated by a combination of microscopic, molecular and spectroscopic approaches. The bacterial community from three rosy-discoloured painting sites was characterized by 16S rRNA gene-based techniques. The eubacterial population was prevalently composed of Actinobacteria, among which Rubrobacter radiotolerans-related bacteria accounted for 63-87% of the 16S rRNA gene pool per sampled site. Archaea, with prevalence of Haloarchaea-related species, were detected in one of the three sites where they accounted for < 0.1% of the total 16S rRNA gene pool. Raman spectroscopy confirmed the identity between R. radiotolerans carotenoids (bacterioruberins) and pigments responsible for colour alteration of frescoes. This investigation provides the first evidence of a causal relationship between heavy contamination by Rubrobacter-related bacterioruberin-producing bacteria and rosy discoloration of ancient wall paintings. PMID- 17922773 TI - Two calcineurin B-like calcium sensors, interacting with protein kinase CIPK23, regulate leaf transpiration and root potassium uptake in Arabidopsis. AB - Calcium signalling involves sensor proteins that decode temporal and spatial changes in cellular Ca2+ concentration. Calcineurin B-like proteins (CBLs) represent a unique family of plant calcium sensors that relay signals by interacting with a family of protein kinases, designated as CBL-interacting protein kinases (CIPKs). In a reverse genetic screen for altered drought tolerance, we identified a loss-of-function allele of CIPK23 as exhibiting a drought-tolerant phenotype. In the cipk23 mutant, reduced transpirational water loss from leaves coincides with enhanced ABA sensitivity of guard cells during opening as well as closing reactions, without noticeable alterations in ABA content in the plant. We identified the calcium sensors CBL1 and CBL9 as CIPK23 interacting proteins that targeted CIPK23 to the plasma membrane in vivo. Expression analysis of the CIPK23, CBL1 and CBL9 genes suggested that they may function together in diverse tissues, including guard cells and root hairs. In addition, expression of the CIPK23 gene was induced by low-potassium conditions, implicating a function of this gene product in potassium nutrition. Indeed, cipk23 mutants displayed severe growth impairment on media with low concentrations of potassium. This phenotype correlates with a reduced efficiency of K+ uptake into the roots. In support of the conclusion that CBL1 and CBL9 interact with and synergistically serve as upstream regulators of CIPK23, the cbl1 cbl9 double mutant, but not the cbl1 or cbl9 single mutants, exhibit altered phenotypes for stomatal responses and low-potassium sensitivity. Together with the recent identification of the potassium channel AKT1 as a target of CIPK23, these results imply that plasma membrane-localized CBL1- and CBL9-CIPK23 complexes simultaneously regulate K+ transport processes in roots and in stomatal guard cells. PMID- 17922774 TI - Population genetics and antibiotic susceptibility of invasive Haemophilus influenzae in Manitoba, Canada, from 2000 to 2006. AB - One hundred and twenty-two isolates of Haemophilus influenzae causing invasive disease were collected in Manitoba, Canada, from 2000 to 2006 and examined for serotype, biotype, sequence type (ST) by multilocus sequence typing and antibiotic susceptibility. Nonserotypeable (NST) isolates accounted for over half of the isolates collected (69 isolates, 56.6%). There were 36 serotype a, five serotype b, two serotype c, one serotype d, four serotype e and five serotype f isolates collected. The 69 NST isolates were found to be very diverse, with isolates representing six biotypes and 45 STs. The serotypeable isolates were more clonal, with each of the serotypes showing little diversity in their biotypes and STs. Of the 122 isolates, 17% were resistant to ampicillin due to beta-lactamase production, 10.7% were resistant to trimethoprim-sulfamethoxazole, 1.6% were resistant to clarithromycin, 2.5% were resistant to amoxicillin clavulanic acid and none was resistant to ciprofloxacin or moxifloxacin. Antibiotic resistance was more common in the NST strains, with 37.7% showing resistance to at least one antibiotic compared to 15% in the serotypeable strains. The results of this study suggest a shift in the epidemiology of invasive H. influenzae infections in the post-Hib vaccine era, and surveillance should include all serotypeable and NST isolates. PMID- 17922775 TI - Neuroleptic malignant syndrome-like state in an epileptic patient with organic brain comorbidity treated with zonisamide and carbamazepine. PMID- 17922777 TI - Reversible facial myoclonus with topiramate therapy for epilepsy. PMID- 17922778 TI - What is the antiseizure activity of acetone due to? PMID- 17922779 TI - Reasons to consider a plasma screen television--photosensitive epilepsy. PMID- 17922769 TI - Viability, diversity and composition of the bacterial community in a high Arctic permafrost soil from Spitsbergen, Northern Norway. AB - The viable and non-viable fractions of the bacterial community in a 2347-year-old permafrost soil from Spitsbergen were subjected to a comprehensive investigation using culture-independent and culture-dependent methods. LIVE/DEAD BacLight staining revealed that 26% of the total number of bacterial cells were viable. Quantitatively, aerobic microcolonies, aerobic colony-forming units and culturable anaerobic bacteria comprised a minor fraction of the total number of viable bacteria, which underlines the necessity for alternative cultivation approaches in bacterial cryobiology. Sulfate reduction was detected at temperatures between -2 degrees C and 29 degrees C while methanogenesis was not detected. Bacterial diversity was high with 162 operational taxonomic units observed from 800 16S rDNA clone sequences. The 158 pure cultures isolated from the permafrost soil affiliated with 29 different bacterial genera, the majority of which have not previously been isolated from permafrost habitats. Most of the strains isolated were affiliated to the genera Cellulomonas and Arthrobacter and several of the pure cultures were closely related to bacteria reported from other cryohabitats. Characterization of viable bacterial communities in permafrost soils is important as it will enable identification of functionally important groups together with the as yet undescribed adaptations that bacteria have evolved for surviving subzero temperatures for millennia. PMID- 17922780 TI - Characteristics and incidence of Clostridium difficile-associated disease in The Netherlands, 2005. AB - During a 2-month period in 2005, 13 laboratories participated in a surveillance study of Clostridium difficile-associated disease (CDAD) in 17 hospitals in The Netherlands. The median incidence rate of CDAD was 16/10 000 patient admissions (2.2/10 000 patient-days) and varied from 1 to 46/10 000 patient admissions according to hospital. In total, 81 patients with CDAD were reported; 49 (61%) patients had nosocomial CDAD, and 29 (36%) patients were admitted to hospital when already suffering from diarrhoea. Two (2%) deaths were attributable to CDAD; both of these patients were admitted with severe community-onset CDAD and were aged >80 years. Among 64 toxinogenic isolates, ten (16%) belonged to PCR ribotype 027 and ten (16%) to PCR ribotype 014. Type 027 was identified in ten patients from one hospital during an unrecognised outbreak. Toxinotyping of the 64 isolates revealed the presence of six different toxinogenic types, with 41 (64%) isolates of toxinotype 0, ten (16%) isolates of toxinotype III, and nine (14%) isolates of toxinotype V. Of the 64 toxinogenic isolates, seven (11%) had a 39-bp deletion in the tcdC gene, 11 (17%) had an 18-bp deletion, and one (1%) had a deletion of c. 44 bp. Genes for binary toxin were present in 21 (33%) of the 64 toxinogenic isolates, mainly associated with toxinotypes III and V. It was concluded that the median CDAD incidence rate of 16/10 000 patient admissions in The Netherlands is considerably lower than that in Canada and the USA, and that the emerging type 027 can spread unnoticed. The high proportion (36%) of CDAD cases with a community onset has important implications for future studies of the epidemiology of CDAD. PMID- 17922782 TI - Laparoscopic radical prostatectomy in the UK: defining and overcoming the obstacles. PMID- 17922783 TI - Urology led ultrasound services--time to focus. PMID- 17922784 TI - Muscarinic receptor antagonists for overactive bladder. AB - Overactive bladder (OAB) is a syndrome characterized by urinary urgency, with or without urgency urinary incontinence, usually with frequency and nocturia. OAB symptoms are often associated with detrusor overactivity (DO). Like OAB symptoms, the prevalence of DO increases with age and can have a neurogenic and/or myogenic aetiology. Bladder outlet obstruction can be a contributing factor in DO, possibly through cholinergic denervation of the detrusor and supersensitivity of muscarinic receptors to acetylcholine, although the prevalence of OAB is similar in men and women across age groups. Acetylcholine is the primary contractile neurotransmitter in the human detrusor, and antimuscarinics exert their effects on OAB/DO by inhibiting the binding of acetylcholine at muscarinic receptors M(2) and M(3) on detrusor smooth muscle cells and other structures within the bladder wall. Worldwide, there are six antimuscarinic drugs currently marketed for the treatment of OAB: oxybutynin, tolterodine, propiverine, trospium, darifenacin, and solifenacin. Each has demonstrated efficacy for the treatment of OAB symptoms, but their pharmacokinetic and adverse event profiles differ somewhat due to structural differences (tertiary vs quaternary amines), muscarinic receptor subtype selectivities, and organ selectivities. Antimuscarinics are generally well tolerated, even in special populations (e.g. men with bladder outlet obstruction, elderly patients, children). The most frequently reported adverse events in clinical studies of antimuscarinics are dry mouth, constipation, headache, and blurred vision; few patients withdraw from clinical trials because of adverse events. Development of an antimuscarinic with functional selectivity for the bladder would reduce the occurrence of antimuscarinic adverse events. The therapeutic potential of several other agents, such as alpha(3)-adrenoceptor agonists, purinergic receptor antagonists, phosphodiesterase inhibitors, neurokinin-1 receptor antagonists, opioids, and Rho kinase inhibitors, is also under investigation for the treatment of OAB. PMID- 17922786 TI - Over-diagnosis of high-grade prostatic intraepithelial neoplasia: a prospective study of 251 cases. AB - OBJECTIVE: To assess the magnitude and causes of over-diagnosis of prostatic intraepithelial neoplasia (PIN), as large differences are reported in the incidence of high-grade PIN, probably because of the variance in diagnosis and interpretation. PATIENTS AND METHODS: Two urological pathologists prospectively reviewed 251 consecutive patients, received in consultation and who were diagnosed and finalized by outside pathologists as having PIN. RESULTS: The diagnosis of PIN was confirmed in 191 patients (incidence of concordance 76.1%, true positive) and refuted in 60 (discordance 23.9%, false positive). The most common histopathological findings misinterpreted as PIN included basal cell hyperplasia, benign epithelium, low-grade PIN, reactive changes, cribriform hyperplasia, atrophy, and post-atrophic hyperplasia. CONCLUSIONS: There is a high rate of over-diagnosis of PIN, usually by misinterpretation of benign mimics. This significant error rate might account for some of the reported differences in the incidence of PIN and the variable predictive accuracy for cancer. PMID- 17922785 TI - Treating patients with overactive bladder syndrome with antimuscarinics: heart rate considerations. AB - In this excellent mini-review, the authors present an extensive and relevant paper on the effect of antimuscarinic agents on the heart. This is without doubt the most detailed and the most reader-friendly paper on this subject, and I am sure that it will help urologists to assist in further educating their patients when prescribing these compounds. PMID- 17922788 TI - Erectile dysfunction, discrepancy between high prevalence and low utilization of treatment options: results from the 'Cottbus Survey' with 10 000 men. AB - OBJECTIVE: To investigate the age-stratified prevalence of erectile dysfunction (ED) and its comorbidities, and to assess the population's knowledge, utilization, and general attitude towards the treatment for ED. SUBJECTS AND METHODS: In all, 10 000 men received a 35-item questionnaire including the International Index of Erectile Function (IIEF) and sociodemographic questions regarding life style, comorbidities, quality of sexual life and knowledge or experience of ED therapy. In all, 3124 responses were included (31.2%), 2499 men lived in well established partnerships and were assessed as the basic study group. RESULTS: In the entire population the prevalence rate of ED was 40.1%. However, although known, medical treatment for ED is used only by a minority of affected men. The prevalence of ED was independently associated with age, peripheral arterial occlusive disease, hypertension, ischaemic heart disease, diabetes mellitus, and liver diseases. Correlations between sexual quality of life (QoL) and ED were statistically significant (P < 0.01) and moderate to strong (absolute values: Spearman's rho 0.35-0.76). Although 96% of the study population knew at least one phosphodiesterase type 5 (PDE5) inhibitor by name, only 53% considered taking the medication and only 9% of the men with ED had had experience with available PDE5 inhibitors. CONCLUSIONS: The sexual QoL was significantly reduced by ED. Despite high levels of awareness and general acceptance of oral medication for ED, experience with PDE5 inhibitors was low. Further investigation is required to evaluate the general impact of ED on sexual QoL and the need or wish for treatment. PMID- 17922787 TI - Does benign prostatic hyperplasia originate from the peripheral zone of the prostate? A preliminary study. AB - OBJECTIVE: To compare the histological characteristics, cell proliferation, apoptosis and biological features in benign prostatic hyperplasia (BPH) in the peripheral (PZ) and transition zone (TZ) of the prostate. PATIENTS AND METHODS: Tissue from BPH in TZ and PZ was obtained from 68 patients undergoing transrectal ultrasonography-guided biopsy and used for both morphometric analysis and immunohistochemical studies. The epithelial, stromal and luminal composition of the tissue was determined using a computer-assisted method for quantitative morphometric analysis. Apoptosis was detected as the apoptotic index (AI) using the TdT dUTP nick-end labelling assay. Cell proliferation was determined as the proliferation index (PI) using Ki-67 immunostaining. The expression of epidermal growth factor receptor (EGFR), transforming growth factor beta1 (TGFbeta1), androgen receptor (AR) and bcl-2 were assessed immunohistochemically. RESULTS: There was no difference in the stroma/epithelium ratio between PZ and TZ hyperplastic nodules (P > 0.05). The mean AI in epithelium was almost identical to the corresponding PI. In stroma, no apoptotic cells were detectable. There was a significantly higher PI and AI in the glandular epithelial cells in PZ hyperplastic than in TZ hyperplastic nodules, but no difference in PI of the stromal cells between PZ and TZ hyperplastic nodules. There was significantly higher expression of TGFbeta1 and lower expression of EGFR and bcl-2 in PZ than TZ hyperplastic nodules (P < 0.05). There was no difference in AR expression between PZ and TZ hyperplastic nodules (P > 0.05). CONCLUSIONS: These results indicate that some hyperplastic nodules in PZ might originate from the PZ, and the formation of these nodules might be modulated in a different way from that in the TZ. PMID- 17922789 TI - Effect of the abnormal bladder when full on upper tract drainage using a combined cystometrogram-(99m)Tc-mercapto-acetyltriglycine renogram: a prospective study. AB - OBJECTIVE: To examine the relevance of bladder volume in patients with chronic hydronephrosis and abnormal bladder function who lose renal function even though the bladder or reservoir pressure is normal (<40 cmH(2)0). PATIENTS AND METHODS: In all, 20 patients (16 male; age range 17-67 years) were studied prospectively; 12 had a reconstructed bladder. All had progressive loss of renal function with a glomerular filtration rate (GFR) of >15 mL/min. The study was in three parts: a baseline (99m)Tc-mercapto-acetyltriglycine (MAG3) scan was performed with an empty bladder, then a combined full bladder cystometrogram (CMG) with (99m)Tc MAG3 study was done supine, and finally repeated in the sitting position. The pressure and volume changes together with the (99m)Tc-MAG3 scintigraphic variables with and without a full bladder were compared. RESULTS: Of the 17 patients with normotensive bladders, 13 (77%) had functional obstruction of the kidneys with a full bladder. In seven this was threshold dependent and occurred at a total mean (range) bladder volume of 348 (135-720) mL for both positions. In the other six patients there was minimal drainage of isotope from the kidneys even with the bladder empty. When the sitting position was compared with supine, gravity was more important for upper tract drainage in the reconstructed bladders. In five patients the detrusor pressure at which drainage began was close to zero. CONCLUSIONS: In 13 of 17 patients (77%) the kidneys failed to drain with a full bladder despite 'normal' detrusor pressures. It remains to be seen whether bladder emptying designed to keep the urine volume below the obstructing threshold volume will prevent further loss of renal function. PMID- 17922790 TI - Ebbing and flowing. PMID- 17922792 TI - Pelvic floor muscle training to improve urinary incontinence after radical prostatectomy: a systematic review of effectiveness. PMID- 17922793 TI - A porcine model of bladder outlet obstruction incorporating radio-telemetered cystometry. PMID- 17922794 TI - Supine valdivia and modified lithotomy position for simultaneous anterograde and retrograde endourological access. PMID- 17922795 TI - Case number and the financial impact of circumcision in reducing prostate cancer. PMID- 17922799 TI - Abstracts from the 2007 Annual Meeting of the American Epilepsy Society. December 1-3, 2007. PMID- 17922797 TI - Laparoscopic orchiopexy. PMID- 17922800 TI - Putative hepatoprotective effects of coffee. PMID- 17922801 TI - Four-day continuous gastric pH monitoring following anti-acid secretory drug administration: cross-over test to assess the early effects. AB - BACKGROUND: There have been few reports that compare the effect of histamine H2 receptor antagonists (H2RAs) and proton pump inhibitors (PPIs) using continuous gastric pH monitoring for a long duration. AIM: To assess the early effects of both drugs on gastric pH using a wireless pH monitoring system. METHODS: The test was conducted by a cross-over test: 10 healthy male volunteers were administered famotidine 20 mg twice a day and lansoprazole 30 mg once a day. Monitoring of gastric pH over four consecutive days was performed using a unique method we have developed that is an elaboration of the Bravo system. RESULTS: The time to reach a pH level of 3 or more with famotidine was significantly shorter than that for lansoprazole. The pH3 holding time ratio of famotidine during the first 4 h of administration was significantly higher than that of lansoprazole. The pH3 holding time ratio on each day from day 1 to day 3 was significantly higher following lansoprazole administration compared with famotidine administration. CONCLUSIONS: Famotidine was shown to act quickly in elevating gastric pH; however, lansoprazole was superior to famotidine in terms of its ability to elevate gastric pH for a long duration. PMID- 17922802 TI - Systematic review and meta-analysis: the clinical and physiological effects of fibre-containing enteral formulae. AB - BACKGROUND: Enteral nutrition can be associated with gastrointestinal side effects and fibre supplementation has been proposed as a means to normalize bowel function. AIM: To evaluate systematically the effects of fibre supplementation of enteral feeds in healthy volunteers and patients both in the hospital and community settings. METHODS: Electronic and manual bibliographic searches were conducted. Controlled studies in adults or children, comparing fibre-supplemented vs. fibre-free formulae given as the sole source of nutrition for at least 3 days, were included. RESULTS: Fifty-one studies (including 43 randomized controlled trials), enrolling 1762 subjects (1591 patients and 171 healthy volunteers) met the inclusion criteria. Fibre supplementation was generally well tolerated. In the hospital setting, the incidence of diarrhoea was reduced as a result of fibre administration (OR 0.68, 95% CI: 0.48-0.96; 13 randomized controlled trials). Meta-regression showed a more pronounced effect when the baseline incidence of diarrhoea was high. In both patients and healthy subjects, fibre significantly reduced bowel frequency when baseline frequency was high and increased it when it was low, revealing a significant moderating effect of fibre. CONCLUSIONS: The review indicates that the fibre-supplemented enteral formulae have important physiological effects and clinical benefits. There is a need to use a consistent approach to undertake more studies on this issue in the community setting. PMID- 17922803 TI - Which knobs fit into which holes in fibrin polymerization? PMID- 17922804 TI - B:b interactions are essential for polymerization of variant fibrinogens with impaired holes 'a'. AB - BACKGROUND: Fibrin polymerization is mediated by interactions between knobs 'A' and 'B' exposed by thrombin cleavage, and holes 'a' and 'b' always present in fibrinogen. The role of A:a interactions is well established, but the roles of knob:hole interactions A:b, B:b or B:a remain ambiguous. OBJECTIVES: To determine whether A:b or B:b interactions have a role in thrombin-catalyzed polymerization, we examined a series of fibrinogen variants with substitutions altering holes 'a': gamma364Ala, gamma364His or gamma364Val. METHODS: We examined thrombin- and reptilase-catalyzed fibrinopeptide release by high-performance liquid chromatography, fibrin clot formation by turbidity, fibrin clot structure by scanning electron microscopy (SEM) and factor (F) XIIIa-catalyzed crosslinking by sodium dodecylsulfate polyacrylamide gel electrophoresis. RESULTS: Thrombin catalyzed fibrinopeptide A release was normal, but fibrinopeptide B release was delayed for all variants. The variant fibrinogens all showed markedly impaired thrombin-catalyzed polymerization; polymerization of gamma364Val and gamma364His were more delayed than gamma364Ala. There was absolutely no polymerization of any variant with reptilase, which exposed only knobs 'A'. SEM showed that the variant clots formed after 24 h had uniform, ordered fibers that were thicker than normal. Polymerization of the variant fibrinogens was inhibited dose-dependently by the addition of either Gly-Pro-Arg-Pro (GPRP) or Gly-His-Arg-Pro (GHRP), peptides that specifically block holes 'a' and 'b', respectively. FXIIIa catalyzed crosslinking between gamma-chains was markedly delayed for all the variants. CONCLUSION: These results demonstrate that B:b interactions are critical for polymerization of variant fibrinogens with impaired holes 'a'. Based on these data, we propose a model wherein B:b interactions participate in protofibril formation. PMID- 17922806 TI - Fibrinolysis in patients with an abdominal aortic aneurysm with special emphasis on rupture and shock. AB - BACKGROUND: A ruptured abdominal aortic aneurysm (AAA) is associated with high mortality. Postoperative complications such as hemorrhage, multiple organ failure, myocardial infarction, and thromboembolism are common. An active and balanced hemostatic system is essential to avoid bleeding as well as thrombosis. When these activities are not properly regulated the patient is at risk of developing either excessive bleeding or thrombosis-related complications. Previous studies have shown a state of activated coagulation in patients with ruptured AAA. However, there are conflicting results regarding the fibrinolytic response. OBJECTIVES: The aim of the present study was to investigate the fibrinolytic state pre-operatively in patients with ruptured and non-ruptured AAA in relation to the clinical outcome with special regard to the influence of shock. METHODS: A prospective study was performed on 95 patients who underwent surgery for a ruptured AAA with shock (n = 43), a ruptured AAA without shock (n = 12), and a non-ruptured AAA (n = 40). Forty-one controls without an aneurysm were matched to the AAA patients according to age, gender and smoking habits. Plasma levels of tissue plasminogen activator antigen (tPAag), and plasminogen activator inhibitor type-1 (PAI-1) were measured as markers of fibrinolytic activity. D dimer, a marker of fibrin turnover, was also measured. RESULTS: D-dimer was significantly higher in patients with a non-ruptured AAA compared with controls without AAA. There were significantly higher levels of D-dimer, tPAag, and PAI-1 in patients operated for ruptured compared with non-ruptured AAA. tPAag was also significantly higher in ruptured AAA patients with shock compared with without shock. No deaths occurred in patients operated on for a non-ruptured AAA or ruptured AAA without shock. There were 12 deaths after repair of a ruptured AAA with shock, of which two patients died from bleeding and the remaining 10 from multiple organ failure and cardiac failure. CONCLUSION: Our results indicate a state of activated coagulation in patients with a non-ruptured AAA, the state being intensified by rupture. The present data show normal fibrinolytic activities in patients with a non-ruptured AAA, but increased systemic fibrinolysis, as demonstrated by elevated tPAag level, in patients with a ruptured AAA. The elevated PAI-1 level indicates a simultaneous inhibition of the systemic fibrinolysis. Furthermore, the hyperfibrinolytic state was reinforced by shock in this study. However, the clinical outcome, with a relatively high incidence of thrombosis-related deaths, indicate a prothrombotic state instead of a hyperfibrinolytic state as a major point of attention in patients with shock as a result of a ruptured AAA. PMID- 17922805 TI - Molecular modeling of the prekallikrein structure provides insights into high molecular-weight kininogen binding and zymogen activation. AB - BACKGROUND: Prekallikrein (PK) plays a central role in the contact system that activates blood coagulation and is involved in the regulation of blood pressure. OBJECTIVES: To provide three-dimensional structural data for PK and rationalize the molecular basis of substrate recognition and zymogen activation. PATIENTS/METHODS: The PK homology model was constructed using the coagulation factor (F) XI crystal structure as a template with the program SWISS-MODEL. RESULTS: The domain organization of the PK apple domains and serine protease is conserved compared to FXI. Surface charge calculations on the PK model revealed that ligand binding to high-molecular-weight kininogen (HK) is predicted to have two key determinants: a pocket within the apple 2 domain and a basic channel formed at the interface of apple domains 1 and 4. A hereditary mutation resulting in PK deficiency (Gly104Arg) and the Lys140 alpha-kallikrein cleavage site both disrupt HK binding and are shown to map to opposite sides of the apple 2 domain pocket. The model also describes the differences in the apple 4 domain that prevents dimer formation in PK vs. FXI. A C-terminal extension in the PK serine protease domain is described as a potential substrate for prolylcarboxypeptidase. CONCLUSIONS: The interaction between PK and HK is mediated by two discrete surfaces formed by the PK A1, A2 and A4 domains with charge likely to be a critical component of the binding. A novel mode of PK activation is postulated to involve prolylcarboxypeptidase cleaving at the C-terminus rather than the activation loop. PMID- 17922807 TI - The effect of exercise on von Willebrand factor and ADAMTS-13 in individuals with type 1 and type 2B von Willebrand disease. AB - BACKGROUND: The effect of exercise on von Willebrand factor (VWF) and ADAMTS-13 levels in individuals with von Willebrand disease (VWD) has never been reported. OBJECTIVES: The aim was to quantify the effect of a standardized exercise protocol on individuals with type 1 and type 2B VWD. PATIENTS/METHODS: Thirty individuals from three groups (10 controls, 11 with type 1 VWD and 9 with type 2B VWD) completed the Standard Bruce Protocol Treadmill Test. A bleeding questionnaire was administered and blood tests were performed pre- and immediately postexercise. The groups were well matched for age, gender and body mass index (BMI). RESULTS: There was a correlation in all groups between the metabolic equivalents (METS) achieved and the degree of change of VWF and FVIII:C levels (P < 0.002, Pearson's correlation). There was a significant postexercise increase in VWF:Ag, VWF:RCo, FVIII:C and activated VWF levels in both the control group and in the type 2B VWD group, but not in the type 1 VWD group. Specific to the type 2B VWD group was an increase in the percentage of high molecular weight multimers (P = 0.022), a decrease in the mean platelet count compared with the other groups (P < 0.001) and an increase in the ADAMTS-13 level (P = 0.001). CONCLUSIONS: There are significant differences in the effects of exercise on individuals with type 1 and type 2B VWD compared with controls. Further clinical studies are necessary to evaluate exercise as a therapeutic option in VWD. PMID- 17922808 TI - Development of sandwich-type ELISAs for the quantification of rat and murine thrombin activatable fibrinolysis inhibitor in plasma. AB - BACKGROUND: Altered plasma levels of thrombin activatable fibrinolysis inhibitor (TAFI) are associated with a large number of pathologies. Rat and murine models are frequently used to study the pathophysiological role of TAFI in vivo but immunological tools to quantify rat and murine TAFI are lacking. OBJECTIVE: The production of monoclonal antibodies (mAb) towards rat TAFI and the development of an ELISA for the quantification of rat and murine TAFI in plasma. METHODS AND RESULTS: Monoclonal antibodies were raised in TAFI-deficient mice towards (activated) recombinant rat TAFI. Pair-wise testing of the mAb revealed three suitable ELISA combinations, namely RT36A3F5/RT30D8-HRP, RT36A3F5/RT82F12-HRP and RT82F12/RT36A3F5-HRP. All three ELISAs are highly specific for rat and murine TAFI. TAFI concentrations in the lower ng mL(-1) range can be determined in plasma samples with a high reproducibility. Comparing TAFI antigen levels measured by these ELISAs with TAFIa activity values determined by activity based assays revealed excellent correlations (R(2) > 0.98). The average antigen levels of 20 individual rat plasma samples were 16 +/- 2 microg mL(-1) using the RT36A3F5-RT30D8-HRP, 12 +/- 2 microg mL(-1) using the RT36A3F5-RT82F12-HRP and 21 +/- 2 microg mL(-1) using the RT82F12-RT36A3F5-HRP ELISA. The determined antigen levels in rat plasma are similar to the levels reported for human plasma. CONCLUSIONS: We developed three highly specific and extremely sensitive sandwich type ELISAs for the quantification of rat and murine TAFI in plasma. The described ELISAs will facilitate in vivo investigation on the pathophysiological role of TAFI. PMID- 17922809 TI - Chemotherapy-induced thrombin generation via procoagulant endothelial microparticles is independent of tissue factor activity. AB - BACKGROUND: Cisplatin-based chemotherapy predisposes cancer patients to thromboembolic events. OBJECTIVES: To investigate whether endothelial damage, via formation of procoagulant endothelial microparticles (EMPs), contributes to cisplatin-related hypercoagulability. METHODS: Cell viability and caspase-3/7 activities were assessed in two endothelial cell (EC) lines [human umbilical vein ECs (HUVECs) and human pulmonary microvascular ECs (HMVEC-Ls)] after exposure to cisplatin (1, 2.5, 5, 10 and 20 microm) for up to 120 h. Counts and procoagulant activity of EMPs were measured by flow cytometry and a thrombin generation assay, respectively. Tissue factor (TF) antigen and TF-dependent procoagulant activity of EMP were determined by enzyme-linked immunosorbent assay and a novel functional assay. RESULTS: By inducing apoptosis, cisplatin dose- and time dependently decreased the viability of confluent HUVECs and HMVEC-Ls. Progression of EC death was accompanied by an increased release of EMPs (relative increase at 20 microm cisplatin for 48 h vs. control: HUVECs 6.5-fold, P < 0.001; HMVEC-Ls 18.4-fold, P < 0.001). EMPs were highly procoagulant (relative increase at 20 microm cisplatin for 48 h vs. control: HUVECs 2.5-fold, P < 0.001; HMVEC-Ls 5.9 fold, P < 0.001). EMP-driven thrombin generation, however, was not dependent on TF: TF expression and TF procoagulant activity levels on microparticles were only marginal and EMP-associated thrombin generation remained unchanged when the extrinsic pathway was blocked by omission of factor VIIa and/or incubation with an anti-human TF antibody. In contrast, blocking of phospholipids by annexin V markedly diminished EMP-associated procoagulant activity. CONCLUSIONS: In vitro, cisplatin induced the release of EMPs that showed TF-independent procoagulant activity. PMID- 17922810 TI - Oxidized low-density lipoprotein-induced expression of ABCA1 in blood monocytes precedes coronary atherosclerosis and is associated with plaque complexity in hypercholesterolemic pigs. AB - BACKGROUND: Elevated oxidized low-density lipoprotein (oxLDL) is associated with atherosclerosis and high cardiovascular risk. Previously, we identified 18 genes in coronary plaque macrophages of hypercholesterolemic pigs that correlated with plaque oxLDL. OBJECTIVE: To determine which of these genes were differentially expressed in blood monocytes and correlated with blood and plaque oxLDL and with plaque complexity. METHODS: RNA expression in monocytes of 27 hypercholesterolemic and 12 control pigs was analyzed with quantitative real-time polymerase chain reaction. RESULTS: Five of 12 genes with detectable expression in monocytes were overexpressed (at P < 0.01 level) in blood monocytes of hypercholesterolemic pigs: ABCA1, SCD, IRF1, SDC2, and TLR2. ABCA1 RNA expression in blood monocytes correlated with blood oxLDL, and its RNA and protein expression was increased prior to atherosclerotic plaque formation. Higher expression of ABCA1 in monocytes was associated with higher plaque complexity and higher plaque oxLDL. Immunostaining of coronary plaques showed the association of ABCA1 with macrophages, lipids, and oxLDL; ABCA1 protein correlated with plaque oxLDL (R(2) = 0.66; P < 0.0001). In THP-1 monocytes, oxLDL induced ABCA1 expression. OxLDL-induced foam cell generation in THP-1 and human monocyte derived macrophages was associated with a further increase of ABCA1 expression. CONCLUSIONS: The increase of ABCA1 in monocytes in association with blood oxLDL prior to atherosclerotic lesion formation and the association of higher ABCA1 with higher plaque complexity suggests that ABCA1 is an early biomarker of atherosclerosis. Studies in humans are warranted. PMID- 17922813 TI - Sweet delivery - sugar translocators as ports of entry for antisense oligodeoxynucleotides in plant cells. AB - Antisense oligodeoxynucleotides (ODNs) are short (12-25 nt long) stretches of single-stranded DNA that may be delivered to a cell, where they hybridize to the cognate mRNA in a sequence-specific manner, thereby inhibiting gene expression. Here we used confocal microscopy to monitor the uptake and trafficking of ODNs in barley tissues. We conclude that uptake of ODNs across the plant plasma membrane is mediated by active transport of mono- or disaccharides through sugar translocators. We demonstrate that sugar transport can deliver ODNs to barley seeds, and that this strategy may be employed to suppress gene activity in endosperm cells by antisense ODN inhibition. We further found that sucrose compared favorably with oligofectamine as a vehicle for ODN delivery to human cells in a low-serum environment. PMID- 17922812 TI - Macrophage colony stimulating factor expression in human cardiac cells is upregulated by tumor necrosis factor-alpha via an NF-kappaB dependent mechanism. AB - INTRODUCTION: Macrophage colony stimulating factor (M-CSF) is a key factor for monocyte and macrophage survival and proliferation. M-CSF has been implicated in cardiac healing and repair after myocardial infarction. METHODS AND RESULTS: We show by immunohistochemistry and Western blotting analysis that M-CSF protein is present in human heart tissue. Cultured human adult cardiac myocytes (HACM) and human adult cardiac fibroblasts (HACF) isolated from human myocardial tissue constitutively express M-CSF. When HACM and HACF were treated with tumor necrosis factor-alpha (TNF-alpha) M-CSF protein production and M-CSF mRNA expression, determined by ELISA or by using RT-PCR, respectively, was significantly increased. To determine a possible role of nuclear factor kappaB (NF-kappaB) and activating protein 1 (AP-1) in M-CSF regulation, blockers to both pathways and an adenovirus overexpressing a dominant negative (dn) form of IkappaB kinase 2 (IKK2) were used. Only the NF-kappaB blocker dimethylfumarate and the dn IKK2, but not januskinase inhibitor-1 (JNK-I), were able to block the TNF-alpha-induced increase in M-CSF production in these cells, suggesting that the induction of M CSF through TNF-alpha is mainly dependent on the activation of the NF-kappaB pathway. The monocyte activation marker CD11b was significantly increased after incubating U937 cells with conditioned medium from HACM or HACF as determined by FACS analysis. CONCLUSIONS: Our in vitro data taken together with our immunohistochemistry data suggest that human cardiac cells constitutively express M-CSF. This expression of M-CSF in the human heart and its upregulation by TNF alpha might contribute to monocyte and macrophage survival and differentiation. PMID- 17922811 TI - Role of the transmembrane domain of glycoprotein IX in assembly of the glycoprotein Ib-IX complex. AB - BACKGROUND: The glycoprotein (GP) Ib-IX complex is critically involved in platelet adhesion to von Willebrand factor and in the initial step of platelet activation. How this complex is assembled is not clear. We previously showed that the transmembrane (TM) domains of the GPIbalpha and GPIbbeta subunits interact and participate in complex assembly. OBJECTIVES AND METHODS: Here, we have investigated the role of the TM and cytoplasmic domains of GPIX in assembly of the GPIb-IX complex, by analyzing the mutational effects on complex expression and assembly in transiently transfected Chinese hamster ovary cells. RESULTS: Replacing the cytoplasmic domain of GPIX with a poly-alanine sequence had little effect on surface expression and structural integrity of the GPIb-IX complex. In contrast, replacing the GPIX TM domain (residues 132-153) with a poly-leucine alanine sequence markedly disrupted complex formation of GPIX with GPIbalpha, interfered with GPIb formation, and decreased surface expression of the host complex. We further analyzed the contributions of a number of GPIX TM residues to complex formation by mutagenesis and found significant roles for Asp135 and several Leu residues. CONCLUSIONS: The TM domain, rather than the cytoplasmic domain, of GPIX plays an important role in expression and assembly of the GPIb-IX complex by interacting with its counterparts of GPIb. These TM domains may form a parallel four-helical bundle structure in the complex. PMID- 17922814 TI - Adaptive differences in gene expression in European flounder (Platichthys flesus). AB - Population structure was previously believed to be very limited or absent in classical marine fishes, but recently, evidence of weakly differentiated local populations has been accumulating using noncoding microsatellite markers. However, the evolutionary significance of such minute genetic differences remains unknown. Therefore, in order to elucidate the relationship between genetic markers and adaptive divergence among populations of marine fishes, we combined cDNA microarray and microsatellite analysis in European flounders (Platichthys flesus). We demonstrate that despite extremely low levels of neutral genetic divergence, a high number of genes were significantly differentially expressed between North Sea and Baltic Sea flounders maintained in a long-term reciprocal transplantation experiment mimicking natural salinities. Several of the differentially regulated genes could be directly linked to fitness traits. These findings demonstrate that flounders, despite little neutral genetic divergence between populations, are differently adapted to local environmental conditions and imply that adaptation in gene expression could be common in other marine organisms with similar low levels of population subdivision. PMID- 17922815 TI - Genetic diversity assessment of anoxygenic photosynthetic bacteria by distance based grouping analysis of pufM sequences. AB - AIM: To assess how completely the diversity of anoxygenic phototrophic bacteria (APB) was sampled in natural environments. METHODS AND RESULTS: All nucleotide sequences of the APB marker gene pufM from cultures and environmental clones were retrieved from the GenBank database. A set of cutoff values (sequence distances 0.06, 0.15 and 0.48 for species, genus, and (sub)phylum levels, respectively) was established using a distance-based grouping program. Analysis of the environmental clones revealed that current efforts on APB isolation and sampling in natural environments are largely inadequate. Analysis of the average distance between each identified genus and an uncultured environmental pufM sequence indicated that the majority of cultured APB genera lack environmental representatives. CONCLUSIONS: The distance-based grouping method is fast and efficient for bulk functional gene sequences analysis. The results clearly show that we are at a relatively early stage in sampling the global richness of APB species. Periodical assessment will undoubtedly facilitate in-depth analysis of potential biogeographical distribution pattern of APB. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first attempt to assess the present understanding of APB diversity in natural environments. The method used is also useful for assessing the diversity of other functional genes. PMID- 17922816 TI - Diversity of Campylobacter coli genotypes in the lower porcine gastrointestinal tract at time of slaughter. AB - AIMS: To compare the genotypes of Campylobacter coli obtained from the rectal and ileal samples of pigs at the time of slaughter. METHODS AND RESULTS: Five animals were sampled following slaughter with ileal contents and anal swabs being taken post-evisceration. Swabs were directly plated onto charcoal cefoperazone desoxycholate agar (CCDA) while ileal contents were enriched in CCDA broth. Twenty isolates were picked from each site sampled and all 200 isolates were Camp. coli. Isolates were genotyped using random amplified polymorphic DNA (22 discrete types) and flaA (11 discrete types). Both methods found that 55% of the genotypes were unique to rectal samples. Only one animal yielded the same flaA type from ileal and rectal samples. CONCLUSIONS: Rectal sampling of pigs yielded a more diverse subset of Camp. coli genotypes than ileal contents, but failed to yield all of the genotypes carried by an individual animal. SIGNIFICANCE AND IMPACT OF THE STUDY: A small sample of pigs carried a very diverse population of Camp. coli genotypes; and sampling of a single site in the gut will recover only part of this population. Hence, any genotyping studies of Camp. coli in pigs must be interpreted with caution, and epidemiological studies could be confounded by the number of Camp. coli genotypes available. PMID- 17922817 TI - Genetic and phenotypic diversity of autochthonous Saccharomyces spp. strains associated to natural fermentation of 'Malvasia delle Lipari'. AB - AIMS: Characterize from both genetic and phenotypic standpoints the indigenous strains of Saccharomyces spp. associated with natural fermentation of 'Malvasia delle Lipari'. METHODS AND RESULTS: A total of 192 yeast isolates were obtained from completed fermentation of a mix of 'Malvasia delle Lipari' (92%) and 'Corinto nero' (8%) grapes in two wineries in Salina Island (Sicily, Italy). Fifty-one Saccharomyces spp. isolates were characterized using ITS-PCR, random amplified polymorphic DNA-PCR and mitochondrial DNA restriction fragment length polymorphism and 12 biotypes were identified. Representative strains of each biotype, tested for their physiological traits, exhibit different killer activity, fermentation vigour, production of hydrogen sulphide and show similar beta-glucosidase and proteolytic activity. CONCLUSIONS: It is possible to cluster in different groups naturally occurring indigenous biotypes of Saccharomyces cerevisiae from 'Malvasia delle Lipari' on the basis of molecular profiles. SIGNIFICANCE AND IMPACT OF THE STUDY: Deeper insight on indigenous wine yeast of a conserved environment. The knowledge gained might offer a contribution to the selection of autochthonous wine yeast as starters for controlled fermentations. PMID- 17922818 TI - Detection of methane and quantification of methanogenic archaea in faeces from young broiler chickens using real-time PCR. AB - AIMS: To detect the presence of methanogens in the faeces of broiler chicks during the first 2 weeks of age. METHODS AND RESULTS: Chicken faecal samples from 120 broiler chicks were incubated for methane gas formation and methanogenic archaea were analysed using real-time PCR. The copy number of the order Methanobacteriales 16S rDNA gene in chicken faeces when the broilers were 3-12 days of age, litter and house flies collected in the bird house ranged from 4.19 to 5.51 log(10) g(-1) wet weight. The number of positive methane culture tubes increased from 25% to 100% as the birds aged. CONCLUSIONS: Methanogens were successfully detected in faecal samples from 3- to 12-day-old broilers, as well as litter and house flies using real-time PCR. The copy number of methanogenic 16S rDNA gene in these samples was also similar to the number observed in litter and house flies. SIGNIFICANCE AND IMPACT OF THE STUDY: The same methanogens consistently appeared in chicken faeces a few days after birth. Detection of the methanogenic bacteria in litter and house flies implicated them as potential environmental sources for methanogen colonization in broiler chicks. PMID- 17922819 TI - Establishment of a human in vitro model of the outer blood-retinal barrier. AB - The outer blood-retinal barrier is composed of a monolayer of retinal pigment epithelium, Bruch's membrane and the choriocapillaris which is fenestrated. Endothelial proliferation and breaching of Bruch's membrane leads to the neovascular form of age-related macula degeneration (ARMD). The aim of this study was to generate an in vitro model that mimics more faithfully the phenotype of the choriocapillaris and the trilayer architecture in vitro. A trilayer culture model was generated with retinal pigment epithelium (ARPE-19) cell cultures on the epithelial surface of amniotic membrane and with human umbilical vein-derived endothelial cells on the other surface. A control model for the effect of retinal pigment epithelium on endothelial changes was generated with corneal epithelial cells replacing the ARPE-19. Both human umbilical vein-derived endothelial and ARPE-19 cells formed confluent monolayers on respective surfaces of the amnion. The human umbilical vein-derived endothelial cells in the trilayer became fenestrated when co-cultured with the ARPE-19 cells, but not with corneal epithelial cells, or when grown as monolayers on the amnion, showing a loss of fidelity of origin in the presence of ARPE-19 cells. These cells also revealed VE cadherin and ZO-1 at cell-cell contacts from 24 h in the trilayer. The tight junctional molecules, occludin and ZO-1, were localized to cell-cell contact regions in the retinal pigment epithelium, both in the monolayer and in the trilayer system. Permeability of the trilayer was tested by using fluorescein and fluorescein-conjugated tracers under flow. At 72 h the trilayer severely restricted transfer of sodium fluorescein (NaF) (ten-fold reduction) whilst transfer of a 4 kDa FITC-conjugated dextran was virtually occluded, confirming a restrictive barrier. Ultrastructural studies showed the retinal pigment epithelium monolayer was polarized with microvilli present on the apical surface. Paracellular clefts showed numerous tight junctional-like appositions, similar to that seen on amnion alone. This study demonstrates that ARPE-19 and human umbilical vein-derived endothelial cells can be co-cultured on the amniotic membrane and that the resultant cross-talk leads to formation of a fenestrated endothelium, whilst maintaining a polarized restrictive epithelial layer. The fenestrated endothelial phenotype achieved in this human in vitro trilayer model is a first and offers an outer-retinal barrier which approaches the in vivo state and has potential for studies into induced junctional disruption, endothelial proliferation and migration: features of ARMD. PMID- 17922820 TI - Purification and partial characterization of a bacteriocin produced by Eikenella corrodens. AB - AIMS: The purpose of this study was to purify and characterize a bacteriocin produced by Eikenella corrodens A32E2. METHODS AND RESULTS: Peptostreptococcus anaerobius ATCC27337 was used as indicator strain in antagonistic assays for bacteriocin-producing E. corrodens A32E2. Protein extraction was influenced by pH and buffer composition. The protein was active in the pH range 6-8. Inhibitory activity was lost by both heating and treatment with proteolytic enzymes and decreased with organic solvents. The substance is rather unstable but maintains 100% of its activity after being exposed to acetone and when stored at -70 degrees C. The antagonistic substance was first precipitated by ammonium sulfate and further partially purified by Mono-Q FPLC and C-18 HPLC. Mass spectrometry analysis showed that the molecular mass was 23 625 Da, and the sequence obtained for the N-terminus was: Met-Asn-Phe-Asp-Glu-Lys-Val-Gly-Lys-Val-X-Phe-Lys-Val-Gly Asp. CONCLUSIONS: The evidence presented in this study supports the idea that an antagonistic substance produced by E. corrodens A32E2 isolated from a periodontal diseased site is a novel bacteriocin, which we designate corrodecin. SIGNIFICANCE AND IMPACT OF THE STUDY: We anticipated that corrodecin might play an important role at the periodontal site. This compound could also be attractive in biotechnological applications as an interesting tool for oral ecosystem control. PMID- 17922821 TI - Characterization of an alkaline active-thiol forming extracellular serine keratinase by the newly isolated Bacillus pumilus. AB - AIMS: The aim of the study was to optimize microbial degradation of keratinous waste and to characterize the alkaline active keratinase showing its biotechnological importance. METHOD AND RESULTS: An extracellular keratinase enzyme was purified from the culture medium of a bacterial isolate and the conditions were optimized. The molecular weight of DEAE-Sepharose-purified keratinase was determined by SDS-PAGE. Instrumental analyses were investigated to study the mechanism of bovine hair hydrolysis. Isolate was identified as Bacillus pumilus based on phenotypic characteristics and 16S rDNA sequence. The optimized condition for its growth was pH 8 and 35 degrees C. The molecular weight of the keratinase was estimated as 65 kDa. Activity inhibition by phenyl methyl sulphonyl fluoride confirmed keratinase as serine protease type. Instrumental analysis revealed the sulphitolysis and proteolysis involved mechanism in bovine hair hydrolysis. CONCLUSION: This study indicates that the isolated keratinase is an alkaline active serine protease with a high degree of activity towards bovine hair. SIGNIFICANCE AND IMPACT OF THE STUDY: This study examines a serine protease with high keratinolytic activity and degradation mechanism for bovine hair. The keratinolytic activity of the isolated strain and its reaction mechanism on bovine hair could show biotechnological potential in the leather industry. PMID- 17922822 TI - Comparative evaluation of the microbial community in biological processes treating industrial and domestic wastewaters. AB - AIMS: Comparison of the microbial composition and process performance between laboratory scale processes treating domestic and vegetable oil wastewaters. METHODS AND RESULTS: Two laboratory scale modified Ludzack-Ettinger processes were operated under similar operating conditions. One process was fed domestic wastewater and the other an industrial wastewater, vegetable oil effluent. Nitrogen removal capacities of the processes were similar. The industrial process exhibited a lower COD removal capacity and oxygen utilization rate, although a greater mixed liquor volatile suspended solids concentration was observed in the industrial process. Fluorescent in situ hybridization (FISH) with probes EUBmix, ALF1b, BET42a, GAM42a and HGC69a revealed that 81% and 72% of total cells stained with 4', 6-diamidino-2-phenylindole (DAPI) within the domestic and industrial processes respectively bound to EUBmix. This indicated a slightly lower Eubacterial population within the industrial process. The alpha-proteobacteria was the dominant community in the industrial process (31% of EUBmix), while the beta-proteobacteria dominated the domestic process (33% of EUBmix). CONCLUSIONS: The findings served to establish a difference in the microbial population between the processes. Therefore, the class alpha-proteobacteria could play a primary role in the degradation of vegetable oil effluent. SIGNIFICANCE AND IMPACT OF THE STUDY: This research will aid in process design and retrofitting of biological processes treating vegetable oil effluent. PMID- 17922824 TI - Optimization of environmental parameters for biodegradation of alpha and beta endosulfan in soil slurry by Pseudomonas aeruginosa. AB - AIM: To determine optimal environmental conditions for achieving biodegradation of alpha- and beta-endosulfan in soil slurries following inoculation with an endosulfan degrading strain of Pseudomonas aeruginosa. METHODS AND RESULTS: Parameters that were investigated included soil texture, soil slurry: water ratios, initial inoculum size, pH, incubation temperature, aeration, and the use of exogenous sources of organic and amino acids. The results showed that endosulfan degradation was most effectively achieved at an initial inoculum size of 600 microl (OD = 0 x 86), incubation temperature of 30 degrees C, in aerated slurries at pH 8, in loam soil. Under these conditions, the bacterium removed more than 85% of spiked alpha- and beta-endosulfan (100 mg l(-1)) after 16 days. Abiotic degradation in noninoculated control medium within same incubation period was about 16%. Biodegradation of endosulfan varied in different textured soils, being more rapid in course textured soil than in fine textured soil. Increasing the soil contents in the slurry above 15% resulted in less biodegradation of endosulfan. Exogenous application of organic acids (citric acid and acetic acid) and amino acids (L-methionine and L-cystein) had stimulatory and inhibitory effects, respectively, on biodegradation of endosulfan. CONCLUSION: The results of this study demonstrated that biodegradation of endosulfan by Ps. aeruginosa in soil sediments enhanced significantly under optimized environmental conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Endosulfan is a commonly used pesticide that can contaminate soil, wetlands and groundwater. Our study demonstrates that bioaugmentation of contaminated soils with an endosulfan degrading bacterium under optimized conditions provides an effective bioremediation strategy. PMID- 17922823 TI - Usefulness of the hydrogen sulfide test for assessment of water quality in Bangladesh. AB - AIM: To evaluate the usefulness of the hydrogen sulfide (H(2)S) test for assessing water quality in Bangladesh. METHODS AND RESULTS: We tested 382 water samples from a variety of sources using locally produced H(2)S test kits and laboratory-based membrane filtration for the detection of Escherichia coli. Compared with membrane filtration, H(2)S tests, when incubated for 24 h, had both a sensitivity and positive predictive value (PPV) of <40% when analysis was restricted to water samples with E. coli levels below 100 colony forming units (CFU) per 100 ml. In contrast, for E. coli levels from 1000 to 9999 CFU per 100 ml, sensitivity was 94% and PPV 88%; specificity was 97% and negative predictive value was 99%. CONCLUSIONS: The hydrogen sulfide test, when incubated at 24 h, is a promising alternative for assessing water quality where E. coli levels may be high. An improved understanding of the incremental impact of contamination level on health is needed to better determine its usefulness. SIGNIFICANCE AND IMPACT OF THE STUDY: The hydrogen sulfide test is inexpensive, easy to use and portable. Its use may allow rapid assessment of water quality in situations where cost or logistics prevent use of other testing methods, such as in remote areas or during flood and other natural disasters. PMID- 17922825 TI - Bioaerosol concentrator performance: comparative tests with viable and with solid and liquid nonviable particles. AB - AIMS: Generally it is more economical to first characterize a concentrator system with nonbiological particles followed by more rigorous bioaerosol testing. This study compares sampling system performance for various particle types and sizes. METHODS AND RESULTS: Performances of five concentrators were characterized with five nonviable and viable laboratory aerosols, although not every concentrator was tested with all aerosol types. For particle sizes less than c. 6 microm aerodynamic diameter, similar efficiencies are obtained for all test particles; however, for larger sizes there is a significant difference between liquid and dry particles. CONCLUSIONS: Aluminium oxide particles provide results over a broad range of sizes with a single test, but the method is less reproducible than other methods. A combination of monodisperse polystyrene spheres and oleic acid droplets provides an accurate representation of the system performance, but ultimately biological particle tests are needed. SIGNIFICANCE AND IMPACT OF THE STUDY: Devices are being developed for concentrating bioaerosol particles in the size range of 1-10 microm aerodynamic diameter and this study provides insight into data quality for different test methodologies. Also, the results show some current concentrators perform quite poorly. PMID- 17922826 TI - Growth, CO2 consumption and H2 production of Anabaena variabilis ATCC 29413-U under different irradiances and CO2 concentrations. AB - AIMS: The objective of this study is to develop kinetic models based on batch experiments describing the growth, CO(2) consumption, and H(2) production of Anabaena variabilis ATCC 29413-U(TM) as functions of irradiance and CO(2) concentration. METHODS AND RESULTS: A parametric experimental study is performed for irradiances from 1120 to 16100 lux and for initial CO(2) mole fractions from 0.03 to 0.20 in argon at pH 7.0 +/- 0.4 with nitrate in the medium. Kinetic models are successfully developed based on the Monod model and on a novel scaling analysis employing the CO(2) consumption half-time as the time scale. CONCLUSIONS: Monod models predict the growth, CO(2) consumption and O(2) production within 30%. Moreover, the CO(2) consumption half-time is an appropriate time scale for analysing all experimental data. In addition, the optimum initial CO(2) mole fraction is 0.05 for maximum growth and CO(2) consumption rates. Finally, the saturation irradiance is determined to be 5170 lux for CO(2) consumption and growth whereas, the maximum H(2) production rate occurs around 10,000 lux. SIGNIFICANCE AND IMPACT OF THE STUDY: The study presents kinetic models predicting the growth, CO(2) consumption and H(2) production of A. variabilis. The experimental and scaling analysis methods can be generalized to other micro-organisms. PMID- 17922827 TI - Boza, a natural source of probiotic lactic acid bacteria. AB - AIMS: To evaluate the probiotic properties of strains isolated from boza, a traditional beverage produced from cereals. METHODS AND RESULTS: The strains survived low pH conditions (pH 3.0), grew well at pH 9.0 and were not inhibited by the presence of 0.3% (w/v) oxbile. Cytotoxicity levels of the bacteriocins, expressed as CC(50), ranged from 38 to 3776 microg ml(-1). Bacteriocin bacST284BZ revealed high activity (EC(50) = 735 microg ml(-1)) against herpes simplex virus type 1. Growth of Mycobacterium tuberculosis was 69% repressed after 5 days in the presence of bacST194BZ. Various levels of auto-cell aggregation and co aggregation with Listeria innocua LMG 13568 were observed. Adhesion of the probiotic strains to HT-29 cells ranged from 18 to 22%. CONCLUSIONS: Boza is a rich source of probiotic lactic acid bacteria. All strains survived conditions simulating the gastrointestinal tract and produced bacteriocins active against a number of pathogens. Adherence to HT-29 and Caco-2 cells was within the range reported for Lactobacillus rhamnosus GG, a well-known probiotic. In addition, the high hydrophobicity readings recorded define the strains as good probiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: Boza contains a number of different probiotic lactic acid bacteria and could be marketed as a functional food product. PMID- 17922828 TI - Diversity of thermophilic populations during thermophilic aerobic digestion of potato peel slurry. AB - AIMS: To study the diversity of thermophiles during thermophilic aerobic digestion (TAD) of agro-food waste slurries under conditions similar to full scale processes. METHODS AND RESULTS: Population diversity and development in TAD were studied by standard microbiological techniques and the processes monitored by standard fermentation procedures. Facultative thermophiles were identified as Bacillus coagulans and B. licheniformis, while obligate thermophiles were identified as B. stearothermophilus. They developed rapidly to peaks of 10(7) to 10(8) in or = 10 microm. At 1 microm, this ligand caused changes in the expression of genes such as TNF, MAF, Rab and Reprimo. At 10 microm, the amidoketone, RG-121150, induced changes in the expression of genes such as v-jun, FBJ and EGR, but was otherwise noninterfering. Of the two steroids tested, 20E did not affect gene expression, but Pon A caused some changes in the expression of endogenous genes. At lower concentrations pharmacologically relevant for gene therapy, intrinsic gene expression effects of ecdysteroids and amidoketones may actually be insignificant. A fortiori, even at 10 microm, the four diacylhydrazine ligands did not cause significant changes in expression of endogenous genes in 293 cells and therefore should have minimum pleiotropic effects when used as ligands for the ecdysone receptor gene switch. PMID- 17922838 TI - The hydroxyproline motif of male sex peptide elicits the innate immune response in Drosophila females. AB - Seminal fluid elicits a variety of physiological and behavioral changes in insect females. In Drosophila melanogaster females, sex peptide (SP) is the major seminal agent eliciting oviposition and reduction of receptivity. But SP also has many other effects; for example, it stimulates food intake, egg production, ovulation, juvenile hormone production and antimicrobial peptide synthesis. Thus, SP very probably has several receptors. To identify putative targets and signaling cascades, we studied the genome-wide regulation of genes by microarray analysis of RNA isolated from females after mating with wild-type males or males lacking SP, respectively. In addition, we studied the effects of SP on the proteome of females. Sex peptide regulates gene activity differentially in the head and in the abdomen. Genes coding for unspecific antimicrobial peptides are specifically transcribed in the abdomen, e.g. the antimicrobial peptide drosocin in epithelial tissues of the female genital tract (oviduct and calyx). Hence, SP elicits a systemic [Peng J, Zipperlen P & Kubli E (2005) Curr Biol15, 1690-1694] and an epithelial immune response. Ectopic expression of SP in the fat body of transgenic virgin females (with subsequent secretion into the hemolymph) does not elicit drosocin synthesis in the genital tract. Thus, the receptors for the stimulation of the systemic and the epithelial responses by SP are compartmentalized. The hydroxyproline (P*) motif of SP, P*TKFP*IP*SP*NP*, is identified as a novel elicitor of the innate immune response. We suggest that SP acts by chemical mimicry of sugar components of the bacterial cell wall. Thus, SP may induce the immune system via pattern recognition receptors. PMID- 17922839 TI - Effect of mutations in the beta5-beta7 loop on the structure and properties of human small heat shock protein HSP22 (HspB8, H11). AB - The human genome encodes ten different small heat shock proteins, each of which contains the so-called alpha-crystallin domain consisting of 80-100 residues and located in the C-terminal part of the molecule. The alpha-crystallin domain consists of six or seven beta-strands connected by different size loops and combined in two beta-sheets. Mutations in the loop connecting the beta5 and beta7 strands and conservative residues of beta7 in alphaA-, alphaB-crystallin and HSP27 correlate with the development of different congenital diseases. To understand the role of this part of molecule in the structure and function of small heat shock proteins, we mutated two highly conservative residues (K137 and K141) of human HSP22 and investigated the properties of the K137E and K137,141E mutants. These mutations lead to a decrease in intrinsic Trp fluorescence and the double mutation decreased fluorescence resonance energy transfer from Trp to bis ANS bound to HSP22. Mutations K137E and especially K137,141E lead to an increase in unordered structure in HSP22 and increased susceptibility to trypsinolysis. Both mutations decreased the probability of dissociation of small oligomers of HSP22, and mutation K137E increased the probability of HSP22 crosslinking. The wild-type HSP22 possessed higher chaperone-like activity than their mutants when insulin or rhodanase were used as the model substrates. Because conservative Lys residues located in the beta5-beta7 loop and in the beta7 strand appear to play an important role in the structure and properties of HSP22, mutations in this part of the small heat shock protein molecule might have a deleterious effect and often correlate with the development of different congenital diseases. PMID- 17922840 TI - Endoplasmic reticulum stress caused by aggregate-prone proteins containing homopolymeric amino acids. AB - Many human proteins have homopolymeric amino acid (HPAA) tracts, but their physiological functions or cellular effects are not well understood. Previously, we expressed 20 HPAAs in mammalian cells and showed characteristic intracellular localization, in that hydrophobic HPAAs aggregated strongly and caused high cytotoxicity in proportion to their hydrophobicity. In the present study, we investigated the cytotoxicity of these aggregate-prone hydrophobic HPAAs, assuming that the ubiquitin proteasome system is impaired in the same manner as other well-known aggregate-prone polyglutamine-containing proteins. Some highly hydrophobic HPAAs caused a deficiency in the ubiquitin proteasome system and excess endoplasmic reticulum stress, leading to apoptosis. These results indicate that the property of causing excess endoplasmic reticulum stress by proteasome impairment may contribute to the strong cytotoxicity of highly hydrophobic HPAAs, and proteasome impairment and the resulting excess endoplasmic reticulum stress is not a common cytotoxic effect of aggregate-prone proteins such as polyglutamine. PMID- 17922841 TI - Effect of oxidative stress and involvement of poly(ADP-ribose) polymerase (PARP) in Dictyostelium discoideum development. AB - Dictyostelium discoideum, a unicellular eukaryote, exhibits multicellularity upon nutrient starvation and is a good model system for developmental studies, and for the study of various signal transduction pathways. Reactive oxygen species at low doses act as signaling molecules; however, at high doses they are known to cause DNA damage that results in the activation of poly(ADP-ribose) polymerase (PARP). We have earlier reported the high resistance of the unicellular stage of D. discoideum to oxidative stress, and we now show the response of this organism to oxidative stress and the role of PARP during development. We used hydroxylamine (HA) to induce in situ generation of H(2)O(2) and monitored the effect of benzamide, a PARP inhibitor, on oxidative stress-induced changes in D. discoideum development. Interestingly, oxidative stress resulted in PARP activation within 5 min that was inhibited by benzamide. Oxidative stress-induced delay in developmental pattern was also partially restored by benzamide. We studied the long-term effects of PARP inhibition under oxidative stress, and our results demonstrated that spores formed under HA stress exhibited significant delay in germination in comparison to benzamide-pretreated HA-stressed cells. However, second-generation cells showed normal development, signifying that PARP inhibition has no deleterious effect on D. discoideum development under oxidative stress. PMID- 17922842 TI - A highly active adipyl-cephalosporin acylase obtained via rational randomization. AB - There is strong interest in creating an enzyme that can deacylate natural cephalosporins such as cephalosporin C in order to efficiently acquire the starting compound for the industrial production of semisynthetic cephalosporin antibiotics. In this study, the active site of the glutaryl acylase from Pseudomonas SY-77 was randomized rationally. Several mutations that were found in previous studies to enhance the activity of the enzyme towards adipyl-7 aminodesacetoxycephalosporanic acid (ADCA) and cephalosporin C have now been combined, and libraries have been made in which random amino acid substitutions at these positions are joined. The mutants were expressed in a leucine-deficient Escherichia coli strain and subjected to growth selection with adipyl-leucine or amino-adipyl-leucine as sole leucine source. The mutants growing on these media were selected and purified, and their hydrolysis activities towards adipyl-7-ADCA and cephalosporin C were tested. Several mutants with highly improved activities towards the desired substrates were found in these rationally randomized libraries. The best mutant was selected from a library of totally randomized residues: 178, 266, and 375. This mutant comprises two mutations, Y178F + F375H, which synergistically improve the catalytic efficiency towards adipyl-7-ADCA 36 fold. The activity of this mutant towards adipyl-7-ADCA is 50% of the activity of the wild-type enzyme towards the preferred substrate glutaryl-7 aminocephalosporanic acid, and therefore the characteristics of this mutant approach those needed for industrial application. PMID- 17922843 TI - Something from nothing: bridging the gap between constraint-based and kinetic modelling. AB - Two divergent modelling methodologies have been adopted to increase our understanding of metabolism and its regulation. Constraint-based modelling highlights the optimal path through a stoichiometric network within certain physicochemical constraints. Such an approach requires minimal biological data to make quantitative inferences about network behaviour; however, constraint-based modelling is unable to give an insight into cellular substrate concentrations. In contrast, kinetic modelling aims to characterize fully the mechanics of each enzymatic reaction. This approach suffers because parameterizing mechanistic models is both costly and time-consuming. In this paper, we outline a method for developing a kinetic model for a metabolic network, based solely on the knowledge of reaction stoichiometries. Fluxes through the system, estimated by flux balance analysis, are allowed to vary dynamically according to linlog kinetics. Elasticities are estimated from stoichiometric considerations. When compared to a popular branched model of yeast glycolysis, we observe an excellent agreement between the real and approximate models, despite the absence of (and indeed the requirement for) experimental data for kinetic constants. Moreover, using this particular methodology affords us analytical forms for steady state determination, stability analyses and studies of dynamical behaviour. PMID- 17922844 TI - Ternary complex formation of pVHL, elongin B and elongin C visualized in living cells by a fluorescence resonance energy transfer-fluorescence lifetime imaging microscopy technique. AB - The tumor suppressor von Hippel-Lindau (VHL) gene product forms a complex with elongin B and elongin C, and acts as a recognition subunit of a ubiquitin E3 ligase. Interactions between components in the complex were investigated in living cells by fluorescence resonance energy transfer (FRET)-fluorescence lifetime imaging microscopy (FLIM). Elongin B-cerulean or cerulean-elongin B was coexpressed with elongin C-citrine or citrine-elongin C in CHO-K1 cells. FRET signals were examined by measuring a change in the fluorescence lifetime of donors and by monitoring a corresponding fluorescence rise of acceptors. Clear FRET signals between elongin B and elongin C were observed in all combinations, except for the combination of elongin B-cerulean and citrine-elongin C. Although similar experiments to examine interaction between pVHL30 and elongin C linked to cerulean or citrine were performed, FRET signals were rarely observed among all the combinations. However, the signal was greatly increased by coexpression of elongin B. These results, together with results of coimmunoprecipitation experiment using pVHL, elongin C and elongin B, suggest that a conformational change of elongin C and/or pVHL was induced by binding of elongin B. The conformational change of elongin C was investigated by measuring changes in the intramolecular FRET signal of elongin C linked to cerulean and citrine at its N- and C-terminus, respectively. A strong FRET signal was observed in the absence of elongin B, and this signal was modestly increased by coexpression of elongin B, demonstrating that a conformation change of elongin C was induced by the binding of elongin B. PMID- 17922845 TI - Expression of the recombinant bacterial outer surface protein A in tobacco chloroplasts leads to thylakoid localization and loss of photosynthesis. AB - Bacterial lipoproteins play crucial roles in host-pathogen interactions and pathogenesis and are important targets for the immune system. A prominent example is the outer surface protein A (OspA) of Borrelia burgdorferi, which has been efficiently used as a vaccine for the prevention of Lyme disease. In a previous study, OspA could be produced in tobacco chloroplasts in a lipidated and immunogenic form. To further explore the potential of chloroplasts for the production of bacterial lipoproteins, the role of the N-terminal leader sequence was investigated. The amount of recombinant OspA could be increased up to ten fold by the variation of the insertion site in the chloroplast genome. Analysis of OspA mutants revealed that replacement of the invariant cysteine residue as well as deletion of the leader sequence abolishes palmitolyation of OspA. Also, decoration of OspA with an N-terminal eukaryotic lipidation motif does not lead to palmitoylation in chloroplasts. Strikingly, the bacterial signal peptide of OspA efficiently targets the protein to thylakoids, and causes a mutant phenotype. Plants accumulating OspA at 10% total soluble protein could not grow without exogenously supplied sugars and rapidly died after transfer to soil under greenhouse conditions. The plants were found to be strongly affected in photosystem II, as revealed by the analyses of temporal and spatial dynamics of photosynthetic activity by chlorophyll fluorescence imaging. Thus, overexpression of OspA in chloroplasts is limited by its concentration-dependent interference with essential functions of chloroplastic membranes required for primary metabolism. PMID- 17922846 TI - Saccharomyces cerevisiae Cox18 complements the essential Sec-independent function of Escherichia coli YidC. AB - Members of the YidC/Oxa1/Alb3 protein family function in the biogenesis of membrane proteins in bacteria, mitochondria and chloroplasts. In Escherichia coli, YidC plays a key role in the integration and assembly of many inner membrane proteins. Interestingly, YidC functions both in concert with the Sec translocon and as a separate insertase independent of the translocon. Mitochondria of higher eukaryotes contain two distant homologues of YidC: Oxa1 and Cox18/Oxa2. Oxa1 is required for the insertion of membrane proteins into the mitochondrial inner membrane. Cox18/Oxa2 plays a poorly defined role in the biogenesis of the cytochrome c oxidase complex. Employing a genetic complementation approach by expressing the conserved region of yeast Cox18 in E. coli, we show here that Cox18 is able to complement the essential Sec-independent function of YidC. This identifies Cox18 as a bona fide member of the YidC/Oxa1/Alb3 family. PMID- 17922847 TI - Substrate recognition by glycoside hydrolase family 74 xyloglucanase from the basidiomycete Phanerochaete chrysosporium. AB - The basidiomycete Phanerochaete chrysosporium produces xyloglucanase Xgh74B, which has the glycoside hydrolase (GH) family 74 catalytic domain and family 1 carbohydrate-binding module, in cellulose-grown culture. The recombinant enzyme, which was heterologously expressed in the yeast Pichia pastoris, had high hydrolytic activity toward xyloglucan from tamarind seed (TXG), whereas other beta-1,4-glucans examined were poor substrates for the enzyme. The existence of the carbohydrate-binding module significantly affects adsorption of the enzyme on crystalline cellulose, but has no effect on the hydrolysis of xyloglucan, indicating that the domain may contribute to the localization of the enzyme. HPLC and MALDI-TOF MS analyses of the hydrolytic products of TXG clearly indicated that Xgh74B hydrolyzes the glycosidic bonds of unbranched glucose residues, like other GH family 74 xyloglucanases. However, viscometric analysis suggested that Xgh74B hydrolyzes TXG in a different manner from other known GH family 74 xyloglucanases. Gel permeation chromatography showed that Xgh74B initially produced oligosaccharides of degree of polymerization (DP) 16-18, and these oligosaccharides were then slowly hydrolyzed to final products of DP 7-9. In addition, the ratio of oligosaccharides of DP 7-9 versus those of DP 16-18 was dependent upon the pH of the reaction mixture, indicating that the affinity of Xgh74B for the oligosaccharides of DP 16-18 is affected by the ionic environment at the active site. PMID- 17922849 TI - The use of genetic testing in hereditary colorectal cancer syndromes: genetic testing in HNPCC, (A)FAP and MAP. AB - This study evaluated the use of genetic testing and time trends in hereditary non polyposis colorectal cancer (HNPCC), (attenuated) familial adenomatous polyposis [(A)FAP] and human MutY homolog (MUTYH) associated polyposis (MAP) families. Eighty-seven families, who were diagnosed with disease-causing mutations between 1995 and 2006, were included in this study. The families consisted of 1547 individuals at risk. Data of these individuals were collected from medical records and family pedigrees. There was considerable interest in genetic testing with test rates of 41% in HNPCC families, 42% in (A)FAP families and 53% in MAP families. The use of genetic testing was associated with age and parenthood. Despite the interest in genetic testing, many risk carriers do not apply for testing. Moreover, time trend analysis showed a decline in test rate in HNPCC families. Studies evaluating the reasons for not testing are needed. Furthermore, a better implementation of genetic testing in clinical practice is desirable. PMID- 17922848 TI - A comparative study of type I and type II tryparedoxin peroxidases in Leishmania major. AB - The genome of Leishmania major, the causative agent of cutaneous leishmaniasis, contains three almost identical genes encoding putative glutathione peroxidases, which differ only at their N- and C-termini. Because the gene homologues are essential in trypanosomes, they may also represent potential drug targets in Leishmania. Recombinant protein for the shortest of these showed negligible peroxidase activity with glutathione as the electron donor indicating that it is not a bone fide glutathione peroxidase. By contrast, high peroxidase activity was obtained with tryparedoxin, indicating that these proteins belong to a new class of monomeric tryparedoxin-dependent peroxidases (TDPX) distinct from the classical decameric 2-Cys peroxiredoxins (TryP). Mass spectrometry studies revealed that oxidation of TDPX1 with peroxides results in the formation of an intramolecular disulfide bridge between Cys35 and Cys83. Site-directed mutagenesis and kinetic studies showed that Cys35 is essential for peroxidase activity, whereas Cys83 is essential for reduction by tryparedoxin. Detailed kinetic studies comparing TDPX1 and TryP1 showed that both enzymes obey saturation ping-pong kinetics with respect to tryparedoxin and peroxide. Both enzymes show high affinity for tryparedoxin and broad substrate specificity for hydroperoxides. TDPX1 shows higher affinity towards hydrogen peroxide and cumene hydroperoxide than towards t-butyl hydroperoxide, whereas no specific substrate preference could be detected for TryP1. TDPX1 exhibits rate constants up to 8 x 10(4) m(-1).s(-1), whereas TryP1 exhibits higher rate constants approximately 10(6) m(-1).s(-1). All three TDPX proteins together constitute approximately 0.05% of the L. major promastigote protein content, whereas the TryPs are approximately 40 times more abundant. Possible specific functions of TDPXs are discussed. PMID- 17922850 TI - A novel duplication in the FMR1 gene: implications for molecular analysis in fragile X syndrome and repeat instability. AB - We have observed a 49 bp tandem duplication adjacent to the triplet repeat of the FMR1 gene and have shown it to occur as a variant in Finland. It affects the primers commonly used in molecular analysis of fragile X syndrome by polymerase chain reaction (PCR) methods. One concern is that females with the full mutation and variant alleles might be missed because of the two PCR products generated by the variant. We suggest that the duplication has arisen by a misalignment of the proximal end of the repeat tract and the non-adjacent GGCGGCGGCGG-sequence located 37 bp upstream and may indicate a mutation hot spot. The discovery of this duplication and the previous observations on deletions associated with full mutations in FMR1 indicate that realignment between the repeat tract and dispersed non-adjacent homologous repetitive sequences may also play a role in repeat instability in fragile X. PMID- 17922851 TI - A case of lethal hypophosphatasia providing new insights into the perinatal benign form of hypophosphatasia and expression of the ALPL gene. AB - Hypophosphatasia is a rare inherited bone disease caused by mutations in the alkaline phosphatase liver-type gene (ALPL) gene, with extensive allelic heterogeneity leading to a range of clinical phenotypes. We report here a patient who died from severe lethal hypophosphatasia, who was compound heterozygous for the mutation c.1133A>T (D361V) and the newly detected missense mutation c791A>G, and whose parents were both healthy. Because the c.1133A>T (D361V) mutation was previously reported to have a dominant-negative effect and to be responsible for the uncommon perinatal benign form of the disease, we studied the expression of the ALPL gene in this family. Analysis at the messenger RNA (mRNA) level, both quantitative and qualitative, showed that the paternal c.1133A>T (D361V) mutation was associated with over-expression of the ALPL gene and that the maternal c.791A>G mutation lead to complete skipping of exon 7. The results provide an explanation of the lethal phenotype in the patient where the two ALPL alleles are non-functional and in the asymptomatic father where over-expression of the normal allele could counteract the effect of the c.1133A>T (D361V) mutation by providing an increased level of normal mRNA. This may also explain the variable expression of hypophosphatasia observed in parents of patients with the perinatal benign form. PMID- 17922852 TI - Low concentrations of doxorubicin sensitizes human solid cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-receptor (R) 2-mediated apoptosis by inducing TRAIL-R2 expression. AB - There is accumulating evidence suggesting that tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-receptor (R) 2 is a promising molecular target for cancer therapy. Therefore, we investigated the effect of chemotherapeutic agents on TRAIL-R2-mediated apoptosis and cytotoxicity in various human solid cancer cells. Treatment of the ACHN human renal cell carcinoma (RCC) cell line with agonistic TRAIL-R2 antibody (lexatumumab) in combination with 5 fluorouracil, vinblastine, paclitaxel, or docetaxel did not overcome resistance to these agents. However, treatment with lexatumumab in combination with doxorubicin had a synergistic cytotoxicity. Synergy was also achieved in two other human RCC cell lines, Caki-1 and Caki-2, and in eight primary RCC cell cultures. Sequential treatment with doxorubicin followed by lexatumumab induced significantly more cytotoxicity than reverse treatment or simultaneous treatment. Low concentrations of doxorubicin (0.1 and 1 microg/mL) significantly increased TRAIL-R2 expression at both the mRNA and protein levels. Furthermore, the combination of doxorubicin and lexatumumab significantly enhanced caspase 8 activity, Bid cleavage, Bcl-xL decrease, release of cytochrome c, and caspase 9 and caspase 3 activity, and induced synergistic apoptosis. The activation of caspases and apoptosis induced with lexatumumab and doxorubicin was blocked by the human recombinant DR5:Fc chimeric protein. In addition, synergistic cytotoxicity was also observed in human prostate, bladder, and lung cancer cells, but was inhibited by the DR5:Fc chimeric protein. These findings suggest that doxorubicin sensitizes solid cancer cells to TRAIL-R2-mediated apoptosis by inducing TRAIL-R2 expression, and that the combination treatment with lexatumumab and doxorubicin might be a promising targeted therapy for cancers, including RCC, prostate, bladder, and lung cancers. PMID- 17922853 TI - Evaluation of costs and morbidity associated with laparoscopic radiofrequency ablation and laparoscopic partial nephrectomy for treating small renal tumours. AB - OBJECTIVE: To compare the costs and morbidity of laparoscopic radiofrequency ablation (LRFA) and laparoscopic partial nephrectomy (LPN) for treating small localized renal tumours. PATIENTS AND METHODS: We retrospectively analysed the outcomes of 88 patients treated at our institution for a renal tumour either by LPN (50) or LRFA (38) between March 2000 and May 2006. Patients with multiple tumours, combined LRFA and LPN, and those who had other simultaneous surgical procedures were excluded. Clinical variables and outcomes were analysed for each patient. Direct cost data were available for 40 patients treated with LPN and 14 with LRFA. Continuous and categorical variables were compared using an independent t-test and chi-square test, respectively. RESULTS: The tumour size was comparable in each group; patients in the LRFA group had more comorbidities (P = 0.01) and a higher overall mortality rate (P = 0.01) but no patient died from cancer. Operative duration, estimated blood loss and length of stay were significantly shorter in the LRFA group but there was no difference in complication rate. LRFA was less costly than LPN ($6103 vs $6808, P = 0.3) but not statistically significantly. The cost savings from the shorter operative duration and length of stay were reduced by the cost of probe. With a median follow-up of 20 months there was no difference in oncological outcome. CONCLUSION: Patients undergoing LRFA tend to be older and have more comorbidities than those treated with LPN. The cost is minimally lower for LRFA, secondary to the added cost of the probe. LRFA might be a good alternative treatment in patients at higher risk of surgical complications, but LPN provides good results when done by an experienced surgeon. PMID- 17922854 TI - Impact of prostate-specific antigen testing on the clinical and pathological outcomes after radical prostatectomy for Gleason 8-10 cancers. AB - OBJECTIVE: To investigate whether the clinical and pathological outcomes after radical retropubic prostatectomy (RRP) have changed since the advent of prostate specific antigen (PSA) testing for patients with Gleason 8-10 cancers. PATIENTS AND METHODS: We identified 584 men treated with RRP between 1988 and 2001 for pathological Gleason 8-10 tumours. Patients were divided for analysis by year of surgery, i.e. early (1988-93), mid (1994-97) and late PSA era (1998-2001). Survival rates after RRP were estimated using the Kaplan-Meier method, and the effect of clinicopathological factors on outcome was analysed using Cox proportional hazard regression models. RESULTS: The median preoperative PSA level decreased from 15 ng/mL in the early to 10 ng/mL in the late PSA era (P < 0.001), while the rate of organ-confined disease increased from 22.9% to 35.1% (P = 0.007). However, the 7-year biochemical recurrence-free (37% vs 45%, P = 0.087) and cancer-specific survival (89% to 91%, P = 0.73) did not change significantly from the early to the late PSA era. Increased preoperative PSA level (P < 0.001), seminal vesicle invasion (P < 0.001) and positive lymph nodes (P = 0.02) were associated with biochemical recurrence. Seminal vesicle invasion (P = 0.005), positive nodes (P < 0.001) and positive surgical margins (P = 0.03) predicted death from cancer. CONCLUSION: Although the pathological features of Gleason 8-10 cancers have become more favourable over the PSA era, survival has not changed. This lack of improvement in clinical outcome probably reflects the inherent biological aggressiveness of these cancers. While RRP provides long-term cancer control in a subset of these patients, continued investigation of multi-modal treatment options is warranted. PMID- 17922855 TI - Prognostic significance of altered p120 ctn expression in bladder cancer. AB - OBJECTIVE: To identify the frequency of change in the expression and localization of p120(ctn) in bladder tumours and its association with clinical outcomes, and to investigate the potential role of p120(ctn) in the migratory and invasive behaviour of bladder carcinoma cells. MATERIALS AND METHODS: In all, 425 superficial tumour specimens (Ta, Tis and T1) and 305 invasive (T2-T4) tumour specimens from 534 patients were assembled in 10 tissue microarrays. P120(ctn) immunostaining was scored for intensity and cellular localization and correlated with clinical variables and survival analysis. Knockdown of p120(ctn) was achieved using small-interference RNA (siRNA) followed by the assessment of migration and invasion behaviour in standard in vitro assays. RESULTS: The expression levels of p120 catenin inversely correlated with pathological tumour stage (P < 0.001), histological grade (P < 0.001), presence of lymphovascular invasion (P = 0.02) but not lymph node (LN) involvement (P = 0.17). Non membranous localization of p120(ctn) correlated with stage (P < 0.001), grade (P < 0.001), lymphovascular invasion (P = 0.04) and LN-positive disease (P = 0.02). A low expression level of p120(ctn) was linked to a poor outcome in cancer specific survival analysis. Knockdown of p120(ctn) using siRNA resulted in a significant reduction in the migration and invasive potential of bladder carcinoma cells. CONCLUSIONS: Our findings suggest that p120(ctn) acts as a prognostic factor in bladder tumours and has a primary role to play in the migratory and invasive behaviour of bladder carcinoma cells. PMID- 17922856 TI - Blood pressure changes during sexual stimulation, ejaculation and midodrine treatment in men with spinal cord injury. AB - OBJECTIVES: To explore the effectiveness of various sources of self-stimulation, including oral midodrine, in triggering ejaculation in men with spinal cord injury (SCI), and to document the systematic variations in blood pressure at ejaculation and consider a revised definition of autonomic dysreflexia. PATIENTS AND METHODS: The study included 62 men with SCI lesions from C2 to L2. Ejaculation potential was assessed with various sources of stimulation, beginning with natural stimulation, followed, if the test was negative, by penile vibrator stimulation (PVS) followed, if the test was again negative, by PVS combined with oral midodrine, started at 5 mg and increased in 5 mg steps up to 25 mg. The success rate of ejaculation was recorded, as were blood pressure (BP) changes measured at baseline and at ejaculation (or on the last trial if the test was negative). Reported sensations were also recorded and compared during positive and negative tests. RESULTS: Overall, 89% of the patients reached ejaculation with one mode or another of stimulation. When patients had a negative result with natural stimulation, 56% were salvaged by PVS, and when PVS was negative, another 22% were salvaged by midodrine combined with PVS. The mean systolic BP increased by 35 mmHg at ejaculation during PVS and by 11 mmHg after midodrine, and a subsequent 29 mmHg at ejaculation during PVS combined with midodrine. By contrast, negative tests showed a relatively stable BP; the difference in changes in BP during positive and negative tests was significant (P < 0.01). Increases in BP during positive tests declined significantly more often within the limits of autonomic dysreflexia than negative tests (P < 0.01). CONCLUSION: These results support the view that most men with SCI can obtain an ejaculation when a wide spectrum of stimulation is used, including natural stimulation, PVS, and PVS combined with oral midodrine. Positive tests were associated with significant increases in BP, in contrast to negative tests, where BP was relatively stable. This suggests that significant changes in BP are required for ejaculation and that insignificant changes are predictive of future failure. As most changes in BP during positive tests also fall within the criterion of autonomic dysreflexia, a revised definition of autonomic dysreflexia should be considered to encourage safe experiences with ejaculation and safe use of midodrine. PMID- 17922857 TI - Pretransplant laparoscopic nephrectomy in adult polycystic kidney disease: a single centre experience. AB - OBJECTIVE: To report our experience with pretransplant laparoscopic nephrectomy in patients with autosomal dominant polycystic kidney disease (ADPKD), as ADPKD often progresses to end-stage renal disease and most azotaemic patients with ADPKD have enlarged kidneys, making graft placement difficult. MATERIAL AND METHODS: We retrospectively reviewed the medical records of 13 patients with renal failure attributable to ADPKD who underwent pretransplant laparoscopic nephrectomy (21 renal units) from August 2002 to December 2006. Five patients had a unilateral nephrectomy, seven had a staged bilateral nephrectomy, and one had a simultaneous bilateral nephrectomy. All patients underwent subsequent living related renal transplantation. The operative duration, haemoglobin decrease, blood transfusion, hospital stay, analgesic requirement and time to receipt of a transplant were compared with those of patients who underwent open pretransplant nephrectomy (14 patients) from 1984 to 2001. RESULTS: Kidneys of a size to interfere with graft placement were the commonest indication for surgery (eight patients). In comparison with open surgery, the mean (SD) hospital stay at 9.26 (2.9) vs 4.86 (0.9) days, analgesic requirement at 320 (120) vs 221 (120.5) mg of tramadol, blood transfusion rate at 1.3 (0.5) vs 0.9 (0.6) units, period to receive a graft kidney at 29.77 (4.6) vs 9.14 (3.38) days, were significantly less with laparoscopy. The complications noted were single instances of splenic capsular tear, pleural tear, sub-acute intestinal obstruction and vena caval injury. CONCLUSION: Pretransplant laparoscopic nephrectomy in patients with ADPKD has all the benefits of minimally invasive surgery such as reduced intraoperative blood loss and minimal postoperative pain leading to early and faster convalescence. These benefits help in decreasing the period between nephrectomy and transplantation. The surgeon needs to have considerable experience in laparoscopy before embarking on laparoscopic pretransplant nephrectomy. PMID- 17922859 TI - Trauma to male genital organs: a 10-year review of 156 patients, including 118 treated by surgery. AB - OBJECTIVE: To assess testicular rupture, scrotal haematoma, penile fracture and penile injury, comparing the prognoses of surgery and conservative management, as trauma to male external genital organs can cause devastating effects on patients and their partners. PATIENTS AND METHODS: We reviewed the medical records of 156 male patients who presented to our emergency centre with trauma to the external genital organs between January 1996 and March 2006. RESULTS: In all, 74 patients had testicular rupture, 32 penile fracture, 26 a penile injury and 24 a scrotal haematoma (mean age 27.8 years). The main cause of trauma was assault (52, 33%). Four of 14 patients with penile trauma who were managed conservatively had complications. Of 20 patients, 17 had a partial orchidectomy and were followed for a month after surgery; scrotal ultrasonography showed three cases of testicular atrophy. The mean hospital stay was less for patients with surgical intervention, at 6.4 days, than for those managed conservatively, at 8.7 days (P < 0.05). A visual analogue pain scale showed less pain in patients who were surgically treated (P < 0.05). CONCLUSION: Prompt surgical intervention is crucial; it should be considered by urologists, and is strongly recommended. Ultrasonography was highly sensitive and specific, and should be used in all patients with trauma to the external genital organs, to aid diagnosis and evaluation before surgery. PMID- 17922858 TI - Can non-malignant biopsy features identify men at increased risk of biopsy detectable prostate cancer at re-screening after 4 years? AB - OBJECTIVES: To identify pathological features in non-malignant sextant prostate needle biopsies and assess their predictive value for detecting prostate cancer on biopsy 4 years later. PATIENTS AND METHODS: We selected and reviewed the biopsy specimens of 121 men that were diagnosed as non-malignant during the first screening round of the European Randomized Study of Screening for Prostate Cancer (ERSPC), Rotterdam section. Of these 61 (50.4%) were positive for cancer during the second round (the result of a matched random sample). The biopsies were indicated by prostate-specific antigen levels of >or= 3.0 ng/mL. Specimens were scored for high-grade prostatic intraepithelial neoplasia (HGPIN), active and chronic inflammation, biopsy core length and glandular core length. The predictive value of the pathological features for detecting prostate cancer after 4 years was assessed. RESULTS: In the first-round biopsies the incidence of HGPIN was 7.1%; there was active inflammation in 22.4% and chronic inflammation in 51.0%. The mean core length was 9.3 mm and mean glandular core length 7.4 mm; the mean total biopsy length (sum of core lengths) was 56.3 mm and mean total glandular length (sum of glandular core lengths) was 44.6 mm. None of the pathological features in the initial round was significantly related to the detection of cancer in the second round. CONCLUSIONS: In this study of non malignant prostate biopsy specimens from a screened population, no pathological features could be identified that were predictive for detecting prostate cancer on biopsy 4 years later. PMID- 17922860 TI - The effect of prostate weight on the outcomes of laparoscopic radical prostatectomy. AB - OBJECTIVE: To evaluate the effect of prostate weight on perioperative, functional and oncological outcomes after laparoscopic radical prostatectomy (LRP). PATIENTS AND METHODS: Between January 2003 and January 2006, 327 patients had LRP by one surgeon, 193 of whom were available for analysis. Patients were stratified into three groups on the basis of pathological prostate weight, i.e. or=75 g. Perioperative, oncological and functional (continence and potency at 1 year) outcomes were compared among the three groups. RESULTS: Of the 193 patients the prostate was or=75 g in 44 (23%); the mean prostate weight was 27, 49 and 98 g in the three subgroups, respectively. At presentation, 144 patients (75%) had T1c disease, 159 (82%) were potent and 187 (97%) were continent. Unilateral nerve-sparing was done in 37 (19%) and bilateral in 114 (59%) patients. The three subgroups were comparable in age, body mass index, preoperative prostate-specific antigen level, preoperative Gleason score, clinical stage, operative duration, length of hospital stay, duration of catheterization, biochemical recurrence and continence after LRP. In the patients with a prostate of 7). In addition, five of eight HRPC samples showed ER-alpha hypermethylation. CONCLUSION: These findings suggest that HMG1 induction as an enhancer of platinum sensitivity is mediated through interaction between oestrogen and ER-alpha. As CpG hypermethylation of the ER-alpha promoter is a frequent event in aggressive prostate cancer, negative conversion of ER alpha methylation is essential to achieve the most beneficial effect when combined chemotherapy of cisplatin with oestrogen is used in patients with prostate cancer. PMID- 17922865 TI - FOURNIER'S gangrene: the development of a classical pathology. AB - Necrotizing fasciitis, especially when termed Fournier's gangrene (FG) when it initiates at the perineum, is a rare but rapidly progressive subcutaneous tissue infection characterized by extensive necrosis. Although it has been known for more than a century and considered as a cause of death, the basic medical principles have not changed for many years. We discuss what is new in the evaluation of this enigmatic pathology and speculate about its clinical metamorphosis. We reviewed reports of FG in the English language. The clinical characteristics of FG have been changing; atypical locations of necrotizing fasciitis, e.g. in the head and neck, and the incidence of patients with FG but no predisposing factors, has been increasing. While the role of anaerobic bacteria in FG is decreasing, that of atypical organisms is increasing, and thus hyperbaric oxygen therapy will probably cease to be a common treatment. We think that FG will not be as likely to cause death in future. PMID- 17922866 TI - Muscarinic and purinergic receptor expression in the urothelium of rats with detrusor overactivity induced by bladder outlet obstruction. AB - OBJECTIVE: To investigate the expression of muscarinic and purinergic receptors in rat urothelium, and changes in their distribution and expression following detrusor overactivity induced by bladder outlet obstruction (BOO). MATERIALS AND METHODS: Thirty Sprague-Dawley rats were divided into control (10) and BOO groups (20). Partial BOO was induced for 3 weeks and the rats assessed by cystometrography. A portion of the bladder was stained using immunofluorescence for M(2) and M(3) muscarinic receptors, and P2X(3) purinergic receptors. The remainder was dissected into bladder urothelium and the smooth muscle layer, and the expression of the receptor proteins analysed by Western blotting. RESULTS: Cystometrography showed a significant decrease in contraction interval and increase in contraction pressure in the BOO group. On immunofluorescence staining, muscarinic and purinergic receptors were localized in both the urothelium and the muscle layer. Immunoreactivity of M(2) and M(3) muscarinic receptors was greater in the urothelium of the BOO group than in the control group; there was a smaller increase in P2X(3) immunoreactivity. On Western blotting, the expression of M(2), M(3) and P2X(3) receptors was increased in the urothelium of the BOO group, and there was increased M(3) receptor expression in the muscle layer of the BOO group. CONCLUSIONS: There were detectable changes in muscarinic and purinergic receptors with bladder overactivity induced by BOO. Our results suggest that changes in urothelium receptor expression could have a role in mediating the afferent sensory responses in the urinary bladder. PMID- 17922867 TI - Glucose transporter-1 expression in renal cell carcinoma and its correlation with hypoxia inducible factor-1 alpha. AB - OBJECTIVE: To evaluate transcription factor hypoxia inducible factor-1 alpha (HIF 1 alpha) activity, by analysing a target gene for HIF-1 alpha, glucose transporter-1 (GLUT-1), using a tissue microarray (TMA) in different types of renal cell carcinoma (RCC, a tumour with a variable clinical course, partly due to angiogenic activity), as angiogenesis is important for tumour progression and metastatic spread, and is activated by hypoxia. PATIENTS AND METHODS: GLUT-1 and HIF-1 alpha expressions were semiquantitatively analysed using immunohistological staining of a prepared TMA, using samples from 187 patients, including 148 with conventional, 26 with papillary and 13 with chromophobe RCC. RESULTS: GLUT-1 staining was found mainly in the cytoplasm. The tumours were subdivided into GLUT -1(LOW) and GLUT-1(HIGH), based on staining intensity. There was a significant difference in GLUT-1 expression between RCC types (P < 0.05). In conventional RCC, GLUT-1 had no correlation with clinicopathological variables. By contrast there was a correlation with tumour stage in papillary RCC. There was an insignificant trend to better survival of patients with GLUT-1(LOW) expression in both conventional and papillary RCC. GLUT-1 correlated significantly (P = 0.008) with HIF-1 alpha. CONCLUSIONS: Most patients with conventional RCC had GLUT 1(HIGH) staining and there was a significant correlation with HIF-1 alpha. In papillary RCC, GLUT-1 expression was associated with stage; GLUT-1 expression was significantly higher in conventional RCC than in papillary and chromophobe RCC. GLUT-1(LOW) in both papillary and conventional RCC appeared to correspond with a better prognosis. PMID- 17922868 TI - Secondary hormonal therapy for prostate cancer: what lies on the horizon? AB - Androgen deprivation therapy with medical or surgical castration is generally the first-line treatment against advanced prostate cancer. Almost invariably, metastatic prostate cancer overcomes testosterone depletion and grows, despite castrate levels of testosterone. Despite advances in cytotoxic chemotherapy, secondary hormonal therapies are often used after the development of castrate resistant prostate cancer. Secondary hormonal therapies either lower the androgen levels further or directly antagonize the androgen receptor in prostate cancer cells. We discuss novel secondary hormonal agents that are under development, which work by either inhibiting androgen synthesis or directly targeting the androgen receptor. PMID- 17922869 TI - Prevalence of renal cysts in a Middle-Eastern population: an evaluation of characteristics and risk factors. AB - OBJECTIVE: To assess the prevalence of renal cysts in a large Middle-Eastern population presenting for a health-screening programme, evaluating cyst characteristics and risk factors for their development. PATIENTS AND METHODS: The reported prevalence of renal cysts detected by ultrasonography (US) in the general population is 5.0-20.8%, and their development has been linked to several factors. The electronic charts of 8551 patients (from eight nations, predominantly Egypt and Yemen) presenting for the 'check-up' programme at the author's institution during 2005 were retrospectively reviewed. The presence and characteristics of renal cysts on abdominal US were noted, as were any associated renal pathologies. Various risk factors were evaluated for renal cyst development, i.e. age, gender, hypertension, diabetes mellitus and serum creatinine levels, and hyperlipidaemia and a history of bilharziasis were also assessed. RESULTS: The prevalence of renal cysts was 4.2%, ranging from 0.6% for patients in their third decade, to a third of those aged >80 years. Cysts were detected in 4.8% of men and 2.8% of women (P < 0.001). The mean serum creatinine level was 1.02 mg/dL in those with cysts and 0.88 mg/dL in those without (P < 0.001). On univariate analysis, hypertension, diabetes and hyperlipidaemia had a significant influence on the occurrence of renal cysts, but not in the multivariate model. Of the 361 patients with renal cysts, 58 (16.1%) had bilateral and 26 (7.2%) had multiple unilateral cysts, with a mean (range) size of 26 (4-104) mm. The vast majority of cysts were classified as Bosniak I simple cysts; seven were Bosniak II and there was one Bosniak IV cyst. Associated renal pathologies included renal stones in 39 patients, hydronephrosis in nine, increased parenchymal echogenicity in 18, small atrophic kidneys in three, haematuria (not associated with other imaging abnormalities) in six, and a renal mass in one patient. CONCLUSIONS: The prevalence of renal cysts detected by US in a health-screened population from the Middle East was 4.2%. Increasing age, male gender and a higher serum creatinine level were significant independent risk factors for cyst development. There was also a relatively high prevalence of associated renal pathologies (increased parenchymal echogenicity and stones). PMID- 17922870 TI - Computer-aided ultrasonography (HistoScanning): a novel technology for locating and characterizing prostate cancer. AB - OBJECTIVE: To assess the extent to which prostate HistoScanning (PHS), a new ultrasound-based technology that uses computer-aided analysis to quantify tissue disorganization induced by malignant processes, can identify and characterize foci of prostate cancer compared with step-sectioned radical prostatectomy (RP) specimens. PATIENTS AND METHODS: Between September 2004 and February 2006, 29 men had PHS before their scheduled RP. A three-dimensional ultrasound raw-data file was acquired, and PHS analysed regions of interest (ROI) corresponding to tissue volumes of approximately 0.04 mL. In 13 men the histology was examined on sections of the whole-mount prostate onto which a grid of 5 x 5 mm squares was applied. On a test set of 14 of the 29 patients, PHS analysis was used before knowing the histology results (blinded data), to predict the maximum tumour diameter, focality, laterality and extraprostatic extension (EPE). RESULTS: Identification and characterization by PHS of the index tumour in the 14 patients in the test set correlated closely (r = 0.95, P < 0.001) with the reference test. The concordance in the attribution of multifocality (present/absent), unilateral/bilateral disease between PHS and histology was 100%. EPE as determined by PHS was attributed to all three pT3a pathological specimens in the blinded paired data. In the same set of data, EPE was attributed to one prostate cancer that on pathological inspection was deemed to be organ-confined (pT2b). CONCLUSIONS: PHS has the potential to identify and characterize prostate cancer foci noninvasively. The precision appears to be sufficient to suggest that PHS might be useful as a triage test for men deemed to be at risk of prostate cancer and who wish to avoid prostate biopsy. PMID- 17922871 TI - Independent assessment of a wide-focus, low-pressure electromagnetic lithotripter: absence of renal bioeffects in the pig. AB - OBJECTIVE: To assess the renal injury response in a pig model treated with a clinical dose of shock waves (SWs) delivered at a slow rate (27 SW/min) using a novel wide focal zone (18 mm), low acoustic pressure (<20 MPa) electromagnetic lithotripter (Xi Xin-Eisenmenger, XX-ES; Xi Xin Medical Instruments Co. Ltd., Suzhou, PRC). MATERIALS AND METHODS: The left kidneys of anaesthetized female pigs were treated with 1500 SWs from either an unmodified electrohydraulic lithotripter (HM3, Dornier MedTech America, Inc., Kennesaw, GA, USA; 18 kV, 30 SW/min) or the XX-ES (9.3 kV, 27 SW/min). Measures of renal function (glomerular filtration rate, GFR, and renal plasma flow) were collected before and after SW lithotripsy, and kidneys were harvested for histological quantification of vascular haemorrhage, expressed as a percentage of the functional renal volume (FRV). A fibre-optic probe hydrophone was used to characterize the acoustic field, and the breakage of gypsum model stones was used to compare the function of the two lithotripters. RESULTS: Kidneys treated with the XX-ES showed no significant change in renal haemodynamic function and no detectable tissue injury. Pigs treated with the HM3 had a modest decline from baseline ( approximately 20%) in both GFR (P > 0.05) and renal plasma flow (P = 0.064) in the treated kidney, but that was not significantly different from the control group. Although most HM3-treated pigs showed no evidence of renal tissue injury, two had focal injury measuring 0.1% FRV, localized to the renal papillae. The width of the focal zone for the XX-ES was approximately 18 mm and that of the HM3 approximately 8 mm. Peak positive pressures at settings used to treat pigs and break model stones were considerably lower for the XX-ES (17 MPa at 9.3 kV) than for the HM3 (37 MPa at 18 kV). The XX-ES required fewer SWs to break stones to completion than did the HM3, with a mean (sd) of 634 (42) and 831 (43) SWs, respectively (P < 0.01). However, conditions were different for these tests because of differences in physical configuration of the two machines. CONCLUSION: The absence of renal injury with the wide focal zone XX-ES lithotripter operated at low shock pressure and a slow SW rate suggests that this lithotripter would be safe when used at the settings recommended for patient treatment. That the injury was also minimal using the Dornier HM3 lithotripter at a slow SW rate implies that the reduced tissue injury seen with these two machines was because they were operated at a slow SW rate. As recent studies have shown stone breakage to be improved when the focal zone is wider than the stone, a wide focal zone lithotripter operated at low pressure and slow rate has the features necessary to provide better stone breakage with less tissue injury. PMID- 17922872 TI - Urethral substitution using autologous lingual mucosal grafts: an experimental study. AB - OBJECTIVE: To investigate the feasibility of urethral substitution using free lingual mucosa grafts (LMGs) in a dog model. MATERIALS AND METHODS: The study included 10 female mongrel dogs in which a 4 x 1 cm(2) section of urethral mucosa was excised. The defect was immediately repaired with a size-matched free LMG harvested from the inferior lateral surface of the tongue. A 12 F urethral catheter was kept inside the urethra for a mean of 7 days. At 3 months after the procedure, the patency of the urethra was assessed by both insertion of a 12 F catheter and by retrograde urethrography. Dogs were killed, the grafted areas excised, and evaluated by gross and histopathological examination. RESULTS: All dogs survived the procedure and there were no tongue complications. One of the 10 dogs developed a slight urethral stricture near the proximal anastomosis. The remaining nine dogs voided spontaneously with no difficulty. Retrograde urethrography showed that no strictures or fistulas had formed. The LMGs shortened by 9.5% after surgery, from a mean (sd) of 4 (0.13) to 3.62 (0.11) cm (statistically significant, P < 0.05). Histological examination showed that the LMGs were well-incorporated into the urethral walls and covered by a keratinized squamous epithelium. Neovascularization was evident beneath the grafts. CONCLUSION: We successfully developed an dog model for free LMGs and showed the feasibility of this approach for urethral substitution. PMID- 17922875 TI - Critical issues in current comparative and cost analyses between retropubic and robotic radical prostatectomy. PMID- 17922874 TI - Patient selection determines the prostate cancer yield of dynamic contrast enhanced magnetic resonance imaging-guided transrectal biopsies in a closed 3 Tesla scanner. AB - OBJECTIVE: To evaluate the cancer yield of transrectal prostate biopsies in a 3-T magnetic resonance imaging (MRI) scanner in patients with elevated prostate specific antigen (PSA) levels and recent negative transrectal ultrasonography (TRUS)-guided prostate biopsies. PATIENTS AND METHODS: Between July 2004 and November 2005, patients with at least one previous negative prostate biopsy within the previous 12 months had MRI-guided biopsy of the prostate in a 3-T MRI scanner. Patients with previous positive biopsies for cancer were excluded. Target selection was based on T2-weighted imaging and dynamic contrast-enhanced (DCE) imaging studies. RESULTS: Thirteen patients were eligible; their median (range) age was 61 (47-74) years and PSA value 4.90 (1.3-12.3) ng/mL. Most patients had one previous negative biopsy (range 1-4). Four patients had a family history of prostate cancer. There were 37 distinct targets based on T2-weighted imaging. Fifteen of 16 distinct DCE abnormalities were co-localized with a target based on T2-weighted imaging. Despite this correlation, only one of 13 patients had a directed biopsy positive for cancer. Including systematic biopsies, two of 13 patients had a biopsy positive for prostate cancer. One patient had prostate intraepithelial neoplasia and one had atypical glands in the specimen. CONCLUSION: The prostate-cancer yield of transrectal biopsies in a 3-T MRI scanner, among patients with recent negative TRUS-guided prostate biopsies, is similar to repeat systematic TRUS-guided biopsy. DCE correlates with T2-imaging but does not appear to improve prostate cancer yield in this population. PMID- 17922873 TI - A phase II trial of cisplatin, fixed dose-rate gemcitabine and gefitinib for advanced urothelial tract carcinoma: results of the Cancer and Leukaemia Group B 90102. AB - OBJECTIVE: To conduct a phase II trial to determine the efficacy of cisplatin, a fixed dose-rate infusion of gemcitabine and gefitinib (an orally active epidermal growth factor receptor tyrosine kinase inhibitor) in patients with advanced urothelial carcinoma. PATIENTS AND METHODS: Eligible patients had previously untreated measurable disease, an Eastern Cooperative Oncology Group performance status of 0-2 and creatinine clearance of >50 mL/min. The treatment regimen consisted of cisplatin 70 mg/m(2) on day 1, gemcitabine 1000 mg/m(2) on day 1 and 8, administered at a fixed dose rate of 10 mg/m(2)/min, given every 3 weeks concurrent with gefitinib 500 mg/day orally for a maximum of six cycles. Maintenance gefitinib 500 mg/day was continued for responding or stable disease. RESULTS: In all, 27 patients were accrued before the study was halted because the dose-limiting toxicity (DLT) exceeded pre-established stopping rules. The DLT events were two grade 5 (one infection, one cardiovascular accident) and three with grade 4 non-haematological toxicity. In 25 evaluable patients there were nine objective responses, for an overall response rate of 36% (95% confidence interval, CI, 18-57%). The median (95% CI) survival time was 11.1 (5.2-35.3) months. CONCLUSION: The combination of cisplatin, fixed dose-rate gemcitabine and gefitinib is active in advanced TCC, although the relative contribution of gefitinib cannot be determined. However, this regimen was associated with excessive toxicity. PMID- 17922876 TI - The scientific assessment of subjective outcome after laparoscopic urological procedures. PMID- 17922877 TI - TRAIL signals to apoptosis in chronic lymphocytic leukaemia cells primarily through TRAIL-R1 whereas cross-linked agonistic TRAIL-R2 antibodies facilitate signalling via TRAIL-R2. AB - Tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF family, which is being developed as an anti-tumour agent due to its selective toxicity to tumour cells, induces apoptosis by binding to two membrane bound receptors, TRAIL-R1 and TRAIL-R2. Clinical trials have been initiated with various preparations of TRAIL as well as agonistic monoclonal antibodies to TRAIL R1 and TRAIL-R2. Previously we reported that prior treatment of primary chronic lymphocytic leukaemia (CLL) cells with histone deacetylase inhibitors was required to sensitize CLL cells to TRAIL and, using various receptor-selective TRAIL mutant ligands, we demonstrated that CLL cells signalled to apoptosis primarily through TRAIL-R1. Some, but not all, agonistic TRAIL-receptor antibodies require cross-linking in order to induce apoptosis. The present study demonstrated that CLL cells can signal to apoptosis through the TRAIL-R2 receptor, but only after cross-linking of the agonistic TRAIL-R2 antibodies, LBY135 and lexatumumab (HGS-ETR2). In contrast, signalling through TRAIL-R1 by receptor-selective ligands or certain agonistic antibodies, such as mapatumumab (HGS-ETR1), occurs in the absence of cross-linking. These results further highlight important differences in apoptotic signalling triggered through TRAIL R1 and TRAIL-R2 in primary tumour cells. Such information is clearly important for the rational optimisation of TRAIL therapy in primary lymphoid malignancies, such as CLL. PMID- 17922879 TI - Survey of members of myeloma UK on biphosphonates-associated jaw osteonecrosis. PMID- 17922878 TI - Chronic lymphocytic leukaemia and radiation: findings among workers at five US nuclear facilities and a review of the recent literature. AB - The aetiology of chronic lymphocytic leukaemia (CLL) is largely unknown. Despite compelling evidence for ionising radiation as a cause of most forms of leukaemia, CLL was not found to be radiogenic in early studies. Herein we describe the recent evidence for causation of CLL by ionising and non-ionising radiation, including a nested case-control study conducted within a cohort of 94 517 US workers at four nuclear weapons facilities and a nuclear naval shipyard. Forty three cases of CLL deaths and 172 age-matched controls were identified with follow-up up to between 1990 and 1996. Radiation exposure from external sources and plutonium (lagged 10 years) was assessed for each worker, based on monitoring records. The excess relative rate (ERR) was estimated for workers receiving elevated doses compared to unexposed workers, controlling for possible risk factors. The ERR per 10 mSv was -0.020 (95% confidence interval: <0, 0.14) based on all exposed workers. However, for workers receiving <100 mSv, the ERR per 10 mSv was 0.20 (-0.035, 0.96). Recent studies of uranium miners and other populations have shown elevations of CLL possibly associated with ionising and non-ionising radiation. New studies should use incident cases and sufficient latency to account for the expected lengthy induction period for CLL. PMID- 17922880 TI - Effects of smoking cessation, acute re-exposure and nicotine replacement in smokers on AIR inhaled insulin pharmacokinetics and glucodynamics. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Only one other study (Becker et al.) has reported on the influence of smoking cessation and smoking resumption on inhaled insulin pharmacokinetics and glucodynamics, concluding that the rapid changes associated with smoking resumption carry the risk for hypoglycaemia and thus should not be used by active smokers. WHAT THIS STUDY ADDS: * This is the first euglycaemic clamp study on the impact of smoking cessation, acute smoking re-exposure and nicotine replacement on AIR((R)) inhaled insulin pharmacokinetics and glucodynamics. * We demonstrate clinically and statistically significant shifts in glucodynamic response to acute re-exposure to a single cigarette, leading us to conclude that active smokers should be advised against inhaled insulin therapy until smoking abstinence is stable. * Additionally, these results are also the first to demonstrate an apparent independent effect of nicotine replacement therapy on insulin exposure and glucodynamic response. AIMS: To explore the effects of smoking cessation and acute smoking re-exposure on the pharmacokinetic (PK) and glucodynamic (GD) profiles of AIR inhaled insulin (AIR Insulin) with or without nicotine replacement therapy (NRT). METHODS: Nondiabetic smokers (n = 24) with normal pulmonary function completed a two-phase (four period), open-label, randomized euglycaemic clamp study. During the initial study phase, subjects underwent glucose clamps following AIR Insulin dosing, shortly after smoking, 8-12 h after smoking, or following subcutaneous insulin lispro shortly after smoking. AIR Insulin PK and GD were again assessed during and after a 4-week smoking-cessation period with or without NRT. In the last study period, subjects smoked one cigarette shortly before final AIR Insulin dosing and glucose clamp, to study the effect of acute smoking re-exposure on inhaled insulin PK and GD. RESULTS: Compared with the preceding active smoking phase, the administration of AIR Insulin in nondiabetic subjects undergoing a 4-week period of smoking abstinence resulted in a decrease in PK and GD of approximately 25% (P = 0.008 for both), an effect which was greater in subjects using NRT. Following rechallenge with a single cigarette (without NRT), GD response to AIR Insulin increased significantly (P = 0.006) towards precessation levels, relative to smoking abstinence. In subjects using NRT, however, the increase in GD was less pronounced. CONCLUSION: Smoking, smoking cessation and acute re-exposure with a single cigarette are associated with clinically significant alterations in AIR Insulin pharmacokinetics and glucodynamics. AIR Insulin should not be used by smokers or those at risk for recidivism. PMID- 17922882 TI - Perforated peptic ulcer and short-term mortality among tramadol users. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Use of nonsteroidal anti-inflammatory drugs (NSAIDs) is a strong risk and prognostic factor for peptic ulcer perforation, and alternative analgesics are needed for high-risk patients. * Pain management guidelines propose tramadol as a treatment option for mild-to-moderate pain in patients at high risk of gastrointestinal side-effects, including peptic ulcer disease. * Tramadol may mask symptoms of peptic ulcer complications, yet tramadol's effect on peptic ulcer prognosis is unknown. WHAT THIS STUDY ADDS: * In this population-based study of 1271 patients hospitalized with peptic ulcer perforation, tramadol appeared to increase mortality at least as much as NSAIDs. * Among users of tramadol, alone or in combination with NSAIDs, adjusted 30-day mortality rate ratios were 2.02 [95% confidence interval (CI) 1.17, 3.48] and 1.32 (95% CI 0.89, 1.95), compared with patients who used neither tramadol nor NSAIDs. AIM: Use of nonsteroidal anti-inflammatory drugs (NSAIDs) increases risk and worsens prognosis for patients with complicated peptic ulcer disease. Therefore, patients who are at high risk of peptic ulcer often use tramadol instead of NSAIDs. Tramadol's effect on peptic ulcer prognosis is unknown. The aim was to examine mortality in the 30 days following hospitalization for perforated peptic ulcer among tramadol and NSAID users compared with non-users. METHODS: The study was based on data on reimbursed prescriptions and hospital discharge diagnoses for the 1993-2004 period, extracted from population-based healthcare databases. All patients with a first-time diagnosis of perforated peptic ulcer were identified, excluding those with previous ulcer diagnoses or antiulcer drug use. Cox regression was used to estimate 30-day mortality rate ratios for tramadol and NSAID users compared with non-users, adjusting for use of other drugs and comorbidity. RESULTS: Of 1271 patients with perforated peptic ulcers included in the study, 2.4% used tramadol only, 38.9% used NSAIDs and 7.9% used both. Thirty-day mortality was 28.7% overall and 48.4% among users of tramadol alone. Compared with the 645 patients who used neither tramadol nor NSAIDs, the adjusted mortality rate in the 30 days following hospitalization was 2.02-fold [95% confidence interval (CI) 1.17, 3.48] higher for the 31 'tramadol only' users, 1.41-fold (95% CI 1.12, 1.78) higher for the 495 NSAID users and 1.32-fold (95% CI 0.89, 1.95) higher for the 100 patients who used both drugs. CONCLUSION: Among patients hospitalized for perforated peptic ulcer, tramadol appears to increase mortality at a level comparable to NSAIDs. PMID- 17922884 TI - Erice statement on drug innovation. PMID- 17922883 TI - Relative lung deposition of salbutamol following inhalation from a spacer and a Sidestream jet nebulizer following an acute exacerbation. AB - What is already known about this subject. Studies have shown that a large volume spacer attached to a metered dose inhaler provides similar bronchodilator effects to nebulized dosing during the management of patients following an acute exacerbation. Due to the high doses used, these effects could be measured at the top of the dose-response relationship and the response limited due to the patient's exacerbation. Although clinical end-points are the gold standard to show comparability, some indication of similar lung deposition is useful to consolidate any claims. What this study adds. The urinary pharmacokinetic method we have used postinhalation provides an index of lung deposition for inhalation methods that can be incorporated into the routine management of patients with an acute exacerbation. This is the first study to identify and compare lung deposition and systemic delivery for inhalation methods within the setting of the routine management of asthma and chronic obstructive pulmonary disease patients following hospitalization due to an acute exacerbation. The study highlights the comparability of the doses for the two inhalation methods evaluated with respect to lung deposition, systemic delivery and bronchodilator response. BACKGROUND: Studies comparing inhalation methods in acute exacerbations have not assessed lung deposition. METHODS: Five 100-mug salbutamol doses were inhaled from a metered dose inhaler plus spacer (MDI + SP) and 5 mg was nebulized (NEB) following acute exacerbation hospitalization. Urinary salbutamol excretion was determined at 30 min (USAL0.5) and over 24 h (USAL24) postinhalation together with forced expiratory volume in 1 s (FEV(1)). RESULTS: The USAL0.5 mean ratio (90% confidence interval) post MDI + SP and NEB [n = 19 asthma, 11 chronic obstructive pulmonary disease (COPD)] was 1.01 (0.81, 1.26). USAL24 was less (P < 0.001) following MDI + SP, whereas FEV(1) was similar. Only a small difference between asthmatics and COPD patients was observed for the MDI + SP in that the USAL0.5 was higher in the asthmatics for the spacer method. CONCLUSION: The relative lung deposition after inhaling 500 mug salbutamol from MDI + SP is similar to 5 mg from a Sidestream nebulizer following an acute exacerbation. PMID- 17922886 TI - Identifying how age and gender influence prescription drug use in a primary health care environment in Catalonia, Spain. AB - What is already known about this subject. Knowledge of prescription patterns in primary health care is an important tool in rational drug therapy. Age and gender are the principal determining factors of cost variability between medical practices, due to drug prescriptions. Age and gender are the principal determining factors of cost variability in relation to the therapeutic group. What this study adds. This study provides specific information on the use of drugs in the primary health care environment of the Catalan Health System, and the differences observed are analyzed with respect to age and gender of the population receiving care. The study shows that there is a high prevalence of drugs in the under 5 year old age group, and also in persons over 54 years of age. The variability found in the cost per patient suggests that adjustment should be made for age in practitioners' prescription evaluation procedures in primary health care in Catalonia. AIMS: To determine the prevalence and usage patterns of prescription drugs according to patients' age and gender, and to identify their relative importance in the prescription costs, in primary health care within the Catalan Health Institute. METHODS: This was a cross-sectional study using computerized pharmacy dispensing records for 5 474 274 members registered, during 2002. Twenty age-gender categories were established. Use of a drug group was defined as filling at least one prescription. The variables studied were age, gender, number of prescriptions and net cost. The prevalence of use, the number of prescriptions and cost issued to each age category were reported. RESULTS: The overall prevalence of drug use was 74.53% (women 80.93%, men 67.84%). This was higher in the group of 0-4 year-olds, and in the >or= 55 year-olds. Age (P < 0.001) produced a statistically more significant effect than gender (P < 0.05). The most used therapeutic groups were analgesics, nonsteroidal anti-inflammatory drugs, antiulcer drugs, anxiolytics, expectorants and mucolytics. The number of prescriptions and costs per patient rose with age and showed great variation in the use of these groups for patients in different age groups. The risk of prescription in women was 23% higher than in men (RR 1.23, 95% CI 1.11, 1.37, P < 0.001). CONCLUSIONS: The majority of subjects were exposed to one or more drugs. The variability in the number of prescriptions and in the prescribing cost per patient between the different age groups suggests that adjustments should be made for age in practitioners' prescription evaluation processes in primary health care in Catalonia. PMID- 17922881 TI - Effect of CYP2C19 polymorphism on nelfinavir to M8 biotransformation in HIV patients. AB - WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: * Nelfinavir is an HIV protease inhibitor, substrate of the transporter P-glycoprotein and metabolized via CYP2C19, CYP3A4 and CYP3A5 enzymes. * Pharmacokinetic studies have shown wide interindividual variability of nelfinavir concentrations, some of this variability perhaps caused by variant drug metabolism or transporter genes. * For CYP3A4*1B and CYP3A5*3 polymorphism, results from three studies are in agreement, showing no difference in nelfinavir concentrations between patients with these different genotypes. * However, for MDR1 and CYP2C19 polymorphism, there have been contradictory studies, showing either no impact on nelfinavir concentration or modified concentrations which could influence virological response. WHAT THIS STUDY ADDS: * Patients with an *1/*2 or *2/*2 genotype for CYP2C19 had a nelfinavir to M8 biotransformation divided by 2 compared with *1/*1 patients. * No evidence of any influence of MDR1 polymorphism on nelfinavir absorption could be detected. AIMS: To evaluate the effect of CYP2C19 polymorphism on nelfinavir and M8 pharmacokinetic variability in human immunodeficiency virus-infected patients and to study the link between pharmacokinetic exposure and short-term efficacy and toxicity. METHODS: Nelfinavir (n = 120) and M8 (n = 119) concentrations were measured in 34 protease inhibitor-naive patients. Two weeks after initiating the treatment, blood samples were taken before, 1, 3 and 6 h after drug administration. Genotyping for CYP3A4, 3A5, 2C19 and MDR1 was performed. A population pharmacokinetic model was developed to describe nelfinavir-M8 concentration time-courses and to estimate interpatient variability. The influence of individual characteristics and genotypes were tested using a likelihood ratio test. Estimated mean (C(mean)), maximal (C(max)) and trough (C(trough)) nelfinavir and M8 concentrations were correlated to short term virological efficacy and tolerance using Spearman nonparametric correlation tests. RESULTS: A one-compartment model with first-order absorption, elimination and metabolism to M8 best described nelfinavir data. M8 was modelled by an additional compartment. Mean pharmacokinetic estimates and the corresponding intersubject variabilities were: absorption rate 0.17 h(-1) (99%), absorption lag time 0.82 h, apparent nelfinavir total clearance 52 l h(-1) (49%), apparent nelfinavir volume of distribution 191 l, M8 elimination rate constant 1.76 h(-1) and nelfinavir to M8 0.39 h(-1) (59%) in *1/*1 patients and 0.20 h(-1) in *1/*2 or *2/*2 patients for CYP2C19*2. Nelfinavir C(mean) was positively correlated to glycaemia and triglyceride increases (P = 0.02 and P = 0.04, respectively). CONCLUSIONS: The rate of metabolism of nelfinavir to M8 was reduced by 50% in patients with *1/*2 or *2/*2 genotype for CYP2C19 compared with those with *1/*1 genotype. PMID- 17922887 TI - Evaluation of medication reviews conducted by community pharmacists: a quantitative analysis of documented issues and recommendations. AB - What is already known about this subject? There is conflicting evidence concerning the potential benefits of pharmacist-led medication review. Little work has been published on the completeness of medication reviews provided by community pharmacists. What this study adds. The 60 community pharmacists taking part in a large randomized controlled trial showed considerable variation in the completeness of the reviews they recorded for intervention patients. Overall, pharmacists recorded only a minority of the potential issues present in these patients. The frequency with which pharmacists recorded issues was not related to key characteristics or to the number of reviews completed. AIMS: To describe issues noted and recommendations made by community pharmacists during reviews of medicines and lifestyle relating to coronary heart disease (CHD), and to identify and quantify missed opportunities for making further recommendations and assess any relationships with demographic characteristics of the pharmacists providing the reviews. METHODS: All issues and recommendations noted by 60 community pharmacists during patient consultations were classified and quantified. Two independent reviewers studied a subsample of cases from every participating pharmacist and identified and classified potential issues from the available data. The findings of the pharmacists and the reviewers were compared. Relevant pharmacist characteristics were obtained from questionnaire data to determine relationships to the proportion of potential issues noted. RESULTS: A total of 2228 issues and 2337 recommendations were noted by the pharmacists in the 738 patients seen, a median of three per patient (interquartile range 2-4). The majority of the recommendations made (1719; 74%) related to CHD. In the subsample of 169 patients (23% of the total), the reviewers identified 1539 potential issues, of which pharmacists identified an average of 33.8% (95% confidence interval 30.1, 36.4). No relationship was found between the proportion of issues noted and potentially relevant factors such as pharmacists' characteristics and their experience of doing reviews. CONCLUSIONS: The majority of issues and recommendations noted by pharmacists related to CHD, although pharmacists recorded only a minority of the issues identified by reviewers. Variation between pharmacists in the completeness of reviews was not explained by review or other relevant experience. PMID- 17922888 TI - Hereditary neuropathy with liability to pressure palsies: description of seven patients without known family history. AB - OBJECTIVES: Hereditary neuropathy with liability to pressure palsies (HNPP) is an inherited disorder resulting in a polyneuropathy with particular involvement at sites of entrapment, and is often underdiagnosed or misdiagnosed. We report findings on seven patients referred for evaluation of focal mononeuropathies or polyneuropathies of undetermined etiology, in whom we established a diagnosis of HNPP. MATERIALS AND METHODS: We retrospectively reviewed clinical, electrophysiological and laboratory data for patients diagnosed with HNPP over a 4-year period at our institution. RESULTS: All patients had transient or recurrent neurological symptoms, some with residual deficits. No patients had a family history of any neuropathy. Electrodiagnostic studies revealed abnormal conduction findings at symptomatic and asymptomatic sites. Testing for the Peripheral Myelin Protein (PMP22) deletion was positive in all patients. CONCLUSIONS: A high index of clinical suspicion and thorough electrodiagnostic evaluation can lead to correct diagnosis of HNPP, despite the absence of a positive family history. PMID- 17922889 TI - Frontal Behavioural Inventory in the differential diagnosis of dementia. AB - OBJECTIVE: To evaluate diagnostic properties of the Frontal Behavioural Inventory (FBI) in patients suffering from different forms of dementia. METHODS: The FBI was administered with other psychometric tests investigating cognitive performances and behavioral scales to the caregivers of 35 patients with the frontal variant of frontotemporal dementia (fv-FTD), 22 patients with Alzheimer's disease (AD) and 15 with vascular dementia (VaD). All patients were comparable for degree of dementia severity and level of executive impairment. RESULTS: The FBI showed high concurrent validity, internal consistency and good inter-rater and test-retest reliability. The discriminant validity was also very high. A new FBI cut-off score of 23 gave 97% sensitivity and 95% specificity in distinguishing fv-FTD from non-FTD patients. Conversely, the Neuropsychiatic Inventory (NPI) score was unable to differentiate fv-FTD from AD. CONCLUSIONS: The FBI is a neurobehavioral tool suitable to distinguish fv-FTD from other forms of dementia also when data from cognitive testing or other behavioral scales fail to support the differential diagnosis. PMID- 17922890 TI - Executive dysfunction in non-demented Parkinson's disease patients with hallucinations. AB - OBJECTIVE: We investigated executive function in Parkinson's disease (PD) patients, and focused on executive dysfunction in PD with hallucinations, but without dementia. METHODS: PD patients were classified by cognitive or neuropsychotic status as PD group, PD with vivid dreaming group, PD with hallucinations group and Parkinson's disease dementia (PDD) group. Psychomotor speed tests, the Stroop test, a verbal fluency test and the Self-rating Depression Scale were performed. RESULTS: The PDD group showed poorer scores in every test compared with the PD group. The PD with hallucinations group showed results similar to those of the PDD group, while the PD with vivid dreaming group was similar to the PD group. CONCLUSIONS: The study suggests that PD patients with hallucinations, not extensive enough to qualify as dementia, already have executive dysfunction similar to that seen in PDD patients. Executive dysfunction may be an important substrate for hallucinations even when dementia is not yet apparent. PMID- 17922891 TI - ACE I/D polymorphism in Korean patients with ischemic stroke and silent brain infarction. AB - OBJECTIVES: Angiotensin-converting enzyme (ACE) polymorphism may play a role in stroke and silent brain infarction (SBI) susceptibility, but the results among the populations studied to date have not been consistent. Thus, we investigated the association between ACE genotypes and ischemic stroke and SBI in Korean patients. SUBJECTS AND METHODS: DNA samples from 237 stroke patients, 264 SBI patients and 234 age-matched controls were amplified using polymerase chain reaction to detect the ACE ins/del (I/D) polymorphism. Genotype was determined by the presence of a 490-bp band (I allele) or a 190-bp band (D allele) in agarose gel electrophoresis. RESULTS: Odds ratios of the I/D and D/D genotypes and the overall (I/D + D/D) for the I/I genotype were significantly different between stroke patients and normal controls. However, there was no significant difference between patients with SBI and controls. CONCLUSIONS: This study is the first report of a significant association between ACE polymorphism and ischemic stroke in the Asian population. Although no consistent associations have been found between ACE polymorphism and stroke in the populations studied to date, the ACE polymorphism may be a genetic determinant of ischemic stroke, at least in Korean patients. PMID- 17922892 TI - Prevalence of narcolepsy-cataplexy in Korean adolescents. AB - BACKGROUND: Narcolepsy typically begins between adolescence and early adulthood causing severe neuropsychiatric impairments, but few prevalence studies are available on adolescent narcoleptics. In the present study, we investigated the prevalence of narcolepsy-cataplexy in adolescents. METHODS: In total 20,407 students, aged 14-19 years, participated in this study. Ullanlinna Narcolepsy Scale (UNS) was applied to all subjects and those with a UNS score of > or =14 were contacted by phone for semi-structured interview. Subjects then suspected of having narcolepsy participated in a laboratory investigation, which included polysomnography and HLA typing, or were interviewed in detail by telephone. RESULTS: Three subjects were finally diagnosed as narcolepsy with cataplexy and seven subjects might be diagnosed as narcolepsy without cataplexy. Among three narcoleptics with cataplexy, two subjects were HLA-DQB1*0602 and DRB1*1501 positive, but one subject had no test of HLA typing. The prevalence of narcolepsy with cataplexy in Korean adolescence was thus determined to be 0.015% (95% confidence interval = 0.0-0.0313%). CONCLUSION: This epidemiologic study is the first of its type on adolescent narcolepsy to use the International Classification of Sleep Disorders, 2nd edition (ICSD-2) diagnostic criteria. Considering those cases with an onset after adolescence were not included, the prevalence of narcolepsy with cataplexy determined in the present study is comparable with that of other studies in adults. PMID- 17922893 TI - Nationwide monitoring of end-of-life care via the Sentinel Network of General Practitioners in Belgium: the research protocol of the SENTI-MELC study. AB - BACKGROUND: End-of-life care has become an issue of great clinical and public health concern. From analyses of official death certificates, we have societal knowledge on how many people die, at what age, where and from what causes. However, we know little about how people are dying. There is a lack of population based and nationwide data that evaluate and monitor the circumstances of death and the care received in the final months of life. The present study was designed to describe the places of end-of-life care and care transitions, the caregivers involved in patient care and the actual treatments and care provided to dying patients in Belgium. The patient, residence and healthcare characteristics associated with these aspects of end-of-life care provision will also be studied. In this report, the protocol of the study is outlined. METHODS/DESIGN: We designed a nationwide mortality follow-back study with data collection in 2005 and 2006, via the nationwide Belgian Sentinel Network of General Practitioners (GPs) i.e. an existing epidemiological surveillance system representative of all GPs in Belgium, covering 1.75% of the total Belgian population. All GPs were asked to report weekly, on a standardized registration form, every patient (>1 year) in their practice who had died, and to identify patients who had died "non suddenly." The last three months of these patients' lives were surveyed retrospectively. Several quality control measures were used to ensure data of high scientific quality. DISCUSSION: In 2005 and 2006, respectively 1385 and 1305 deaths were identified of which 66% and 63% died non-suddenly. The first results are expected in 2007. Via this study, we will build a descriptive epidemiological database on end-of-life care provision in Belgium, which might serve as baseline measurement to monitor end-of-life care over time. The study will inform medical practice as well as healthcare authorities in setting up an end-of-life care policy. We publish the protocol here to inform others, in particular countries with analogue GP surveillance networks, on the possibilities of performing end-of life care research. A preliminary analysis of the possible strengths, weaknesses and opportunities of our research is outlined. PMID- 17922894 TI - Using death certificate data to study place of death in 9 European countries: opportunities and weaknesses. AB - BACKGROUND: Systematic and reliable epidemiological information at population level, preferably cross-national, is needed for an adequate planning of (end-of life) health care policies, e.g. concerning place of death, but is currently lacking. This study illustrates opportunities and weaknesses of death certificate data to provide such information on place of death and associated factors in nine European countries (seven entire countries and five regions). METHODS: We investigated the possibility and modality of all partners in this international comparative study (BE, DK, IT, NL, NO, SE, UK) to negotiate a dataset containing all deaths of one year with their national/regional administration of mortality statistics, and analysed the availability of information about place of death as well as a number of clinical, socio-demographic, residential and healthcare system factors. RESULTS: All countries negotiated a dataset, but rules, procedures, and cost price to get the data varied strongly between countries. In total, about 1.1 million deaths were included. For four of the nine countries not all desired categories for place of death were available. Most desired clinical and socio-demographic information was available, be it sometimes via linkages with other population databases. Healthcare system factors could be made available by linking existing healthcare statistics to the residence of the deceased. CONCLUSION: Death certificate data provide information on place of death and on possibly associated factors and confounders in all studied countries. Hence, death certificate data provide a unique opportunity for cross national studying and monitoring of place of death. However, modifications of certain aspects of death certificate registration and rules of data-protection are perhaps required to make international monitoring of place of death more feasible and accurate. PMID- 17922895 TI - Pharmacokinetics of recombinant human growth hormone administered by cool.click 2, a new needle-free device, compared with subcutaneous administration using a conventional syringe and needle. AB - BACKGROUND: Growth hormone (GH) is used to treat growth hormone deficiency (GHD, adult and paediatric), short bowel syndrome in patients on a specialized diet, HIV-associated wasting and, in children, growth failure due to a number of disorders including Turner's syndrome and chronic renal failure, and in children born small for gestational age. Different brands and generic forms of recombinant human growth hormone (r-hGH) are approved for varying indications in different countries. New ways of administering GH are required because the use of a needle and syringe or a device where a patient still has to insert the needle manually into the skin on a daily basis can lead to low adherence and sub-optimal treatment outcomes. The objective of this study was to assess the relative bioavailability of r-hGH (Saizen, Merck Serono) administered by a new needle-free device, cool.click 2, and a standard needle and syringe. METHODS: The study was performed with 38 healthy volunteers who underwent pituitary somatotrope cell down-regulation using somatostatin, according to a randomized, two-period, two sequence crossover design. Following subcutaneous administration of r-hGH using cool.click 2 or needle and syringe, pharmacokinetic parameters were analysed by non-compartmental methods. Bioequivalence was assessed based on log-transformed AUC and C(max) values. RESULTS: The 90% confidence intervals for test/reference mean ratio of the plasma pharmacokinetic variables Cmax and AUC(0-inf) were 103.7 118.3 and 97.1-110.0, respectively, which is within the accepted bioequivalence range of 80-125%. r-hGH administered by cool.click 2 is, therefore, bioequivalent to administration by needle and syringe with respect to the rate and extent of GH exposure. Treatment using cool.click 2 was found to be well tolerated. With cool.click 2 the tmax was less (3.0 hours) than for needle and syringe delivery (4.5 hours), p = 0.002 (Friedman test), although this is unlikely to have any clinical implications. CONCLUSION: These results demonstrate that cool.click 2 delivers subcutaneous r-hGH exposure that is bioequivalent to the conventional mode of injection. The new device has the additional advantage of being needle free, and should help to increase patient adherence and achieve good therapeutic outcomes from r-hGH treatment. PMID- 17922896 TI - Tripartite chimeric pseudogene from the genome of rice blast fungus Magnaporthe grisea suggests double template jumps during long interspersed nuclear element (LINE) reverse transcription. AB - BACKGROUND: A systematic survey of loci carrying retrotransposons in the genome of the rice blast fungus Magnaporthe grisea allowed the identification of novel non-canonical retropseudogenes. These elements are chimeric retrogenes composed of DNA copies from different cellular transcripts directly fused to each other. Their components are copies of a non protein-coding highly expressed RNA of unknown function termed WEIRD and of two fungal retrotransposons: MGL and Mg SINE. Many of these chimeras are transcribed in various M. grisea tissues and during plant infection. Chimeric retroelements with a similar structure were recently found in three mammalian genomes. All these chimeras are likely formed by RNA template switches during the reverse transcription of diverse LINE elements. RESULTS: We have shown that in M. grisea template switching occurs at specific sites within the initial template RNA which contains a characteristic consensus sequence. We also provide evidence that both single and double template switches may occur during LINE retrotransposition, resulting in the fusion of three different transcript copies. In addition to the 33 bipartite elements, one tripartite chimera corresponding to the fusion of three retrotranscripts (WEIRD, Mg-SINE, MGL-LINE) was identified in the M. grisea genome. Unlike the previously reported two human tripartite elements, this fungal retroelement is flanked by identical 14 bp-long direct repeats. The presence of these short terminal direct repeats demonstrates that the LINE enzymatic machinery was involved in the formation of this chimera and its integration in the M. grisea genome. CONCLUSION: A survey of mammalian genomic databases also revealed two novel tripartite chimeric retroelements, suggesting that double template switches occur during reverse transcription of LINE retrotransposons in different eukaryotic organisms. PMID- 17922898 TI - Oxcarbazepine as monotherapy of acute mania in insufficiently controlled type-1 diabetes mellitus: a case-report. AB - BACKGROUND: Type-1 diabetes mellitus (DM) is a lifelong serious condition which often renders the application of standard treatment options for patients' comorbid conditions, such as bipolar disorder I, risky - especially for acute manic episodes. We present such a case whereby the application of standard anti manic treatments would have jeopardized a patient whose physical condition was already compromised by DM. METHODS: We report the case of a 55-year-old female with a history of type-1 DM since the age of 11, and severe ocular and renal vascular complications thereof. While on the waiting list for pancreatic islet cell transplantation, she developed a manic episode that proved recalcitrant to a treatment with gabapentin, lorazepam and quetiapine. Moreover, her mental state affected adversely her already compromised glycemic control, requiring her psychiatric hospitalization. Her psychotropic medication was almost discontinued and replaced by oxcarbazepine (OXC) up to 1800 mg/day for 10 days. RESULTS: The patient's mental state improved steadily and on discharge, 3 weeks later, she showed an impressive improvement rate of over 70% on the YMRS. Moreover, she remains normothymic 6 months after discharge, with OXC at 1200 mg/day. CONCLUSION: Standard prescribing guidelines for acute mania recommend a combination of an antipsychotic with lithium or, alternatively, a combination of an antipsychotic with valproate or carbamazepine. However, in our case, administration of lithium was at least relatively contra-indicated because of patient's already compromised renal function. Furthermore, antipsychotics increase glucose levels and thus were also relatively contra-indicated. Moreover, the imminent post-transplantation immunosuppressant treatment with immuno modulating medicines also contra-indicated both valproate and carbamazepine. Despite the severe methodological limitations of case reports in general, the present one suggests that OXC as monotherapy might be both safe and efficacious in the treatment of acute mania in patients with early-onset type-1 DM, whose already compromised physical condition constitutes an absolute or relative contra indication for the administration of standard treatments, though there are no, as yet, randomized clinical trials attesting to its efficacy unambiguously. PMID- 17922899 TI - Pharmacokinetics, safety and efficacy from randomized controlled trials of 1 and 2 mg nicotine bitartrate lozenges (Nicotinell). AB - BACKGROUND: The use of nicotine replacement therapy (NRT) can almost double the chances of success for smokers to quit. Nevertheless, there is still a considerable number of cessation attempts that are made without any treatment. This novel oral formulation, (lozenge containing nicotine bitartrate dihydrate) has been developed to enlarge the offer for efficient smoking cessation drug therapies, assuming that increasing treatment options will bring more smokers to find the support they personally need to stop smoking. METHODS: Three pharmacokinetic (PK), one safety and two efficacy studies were carried out with Nicotinell lozenges. PK trials were: (1) a single-dose, three-way crossover study comparing 1 and 2 mg lozenges with 2 mg nicotine gum; (2) a multiple-dose, two way crossover study comparing 1 mg lozenge with 2 mg gum; (3) a multiple-dose, three-way crossover study comparing 1 and 2 mg lozenges with 4 mg gum. Safety trial: (4) a single dose study to assess the safety of swallowing up to 12 lozenges containing 1 mg nicotine. Efficacy trials: two efficacy studies in (5) France and (6) the USA, including more than 900 smokers followed-up for up to one year, conducted with the 1 mg lozenge. RESULTS: The results of the individual PK trials showed that the 1 mg Nicotinell lozenge is bioequivalent to 2 mg polacrilex gum, as demonstrated by similar blood PK parameters (tmax, Cmax, AUC). The 2 mg lozenge was found to deliver quantities of nicotine that were intermediate between those delivered by 2 and 4 mg polacrilex gum. The short-term efficacy of the 1 mg lozenge in comparison with placebo was also demonstrated with significantly more subjects continuously abstinent from smoking with active lozenges on week 6 in two different populations: moderate to heavy smokers (FTND between 4 and 7) OR = 1.72 [95% CI: 1.05-2.80]; heavy to very heavy smokers (FTND 6 and over) OR = 2.87 [95% CI: 1.18-6.97]. Nicotinell lozenges were found to be safe with mainly mild and reversible adverse events. The safety of the 1 mg lozenge formulation, even when misused was also demonstrated. CONCLUSION: The data presented in this review demonstrate high nicotine bioavailability, excellent safety profile and proven short-term efficacy of Nicotinell lozenges. At nominal equivalent doses 1 and 2 mg Nicotinell lozenges were shown to deliver larger amounts of bioavailable nicotine compared to the nicotine polacrilex gum. According to the data developed here, the systemic exposure to nicotine could be ranked: 4 mg polacrilex gum > 2 mg Nicotinell lozenge > 1 mg Nicotinell lozenge = 2 mg polacrilex gum.Adverse events observed during the clinical trials were mild or moderate in severity, transient and completely reversible. With respect to efficacy in smoking cessation, significantly higher continuous abstinence rates were achieved with lozenge compared to placebo. In conclusion, Nicotinell lozenges offer a valuable addition to the therapeutic armamentarium available for smoking cessation. PMID- 17922897 TI - Transcriptional profiling of inductive mesenchyme to identify molecules involved in prostate development and disease. AB - BACKGROUND: The mesenchymal compartment plays a key role in organogenesis, and cells within the mesenchyme/stroma are a source of potent molecules that control epithelia during development and tumorigenesis. We used serial analysis of gene expression (SAGE) to profile a key subset of prostatic mesenchyme that regulates prostate development and is enriched for growth-regulatory molecules. RESULTS: SAGE libraries were constructed from prostatic inductive mesenchyme and from the complete prostatic rudiment (including inductive mesenchyme, epithelium, and smooth muscle). By comparing these two SAGE libraries, we generated a list of 219 transcripts that were enriched or specific to inductive mesenchyme and that may act as mesenchymal regulators of organogenesis and tumorigenesis. We identified Scube1 as enriched in inductive mesenchyme from the list of 219 transcripts; also, quantitative RT-PCR and whole-mount in situ hybridization revealed Scube1 to exhibit a highly restricted expression pattern. The expression of Scube1 in a subset of mesenchymal cells suggests a role in prostatic induction and branching morphogenesis. Additionally, Scube1 transcripts were expressed in prostate cancer stromal cells, and were less abundant in cancer associated fibroblasts relative to matched normal prostate fibroblasts. CONCLUSION: The use of a precisely defined subset of cells and a back-comparison approach allowed us to identify rare mRNAs that could be overlooked using other approaches. We propose that Scube1 encodes a novel stromal molecule that is involved in prostate development and tumorigenesis. PMID- 17922900 TI - Catching the news: Processing strategies in listening to dialogs as measured by ERPs. AB - BACKGROUND: The online segmentation of spoken single sentences has repeatedly been associated with a particular event-related brain potential. The brain response could be attributed to the perception of major prosodic boundaries, and was termed Closure Positive Shift (CPS). However, verbal exchange between humans is mostly realized in the form of cooperative dialogs instead of loose strings of single sentences. The present study investigated whether listeners use prosodic cues for structuring larger contextually embedded utterances (i.e. dialogs) like in single sentence processing. METHODS: ERPs were recorded from listeners (n = 22) when presented with question-answer dialogs in German. The prosody of the answer (target sentence) either matched the context provided by a question or did not match the context question. RESULTS: CPS responses to the processing of the target sentences are elicited, first, when listeners encounter information comprising 'novelties', i.e. information not mentioned in the preceding question but facts corrected between context and target. Thereby it is irrelevant whether the actual prosody of the target sentence is in congruence with the informative status or not. Second, when listeners encounter target sentences which do not convey any novelties but only previously 'given' already known information, the structuring of the speech input is driven by prosody again. The CPS is then elicited when listeners perceive major prosodic boundaries similar as for the processing of context-free single sentences. CONCLUSION: The study establishes a link between the on-line structuring of context-free (single sentences) and context-embedded utterances (dialogs) as measured by ERPs. Moreover, the impact of prosodic phrasing and accentuation on the perception of spoken utterances on and beyond sentence level is discussed. PMID- 17922901 TI - The BLISS cluster randomised controlled trial of the effect of 'active dissemination of information' on standards of care for premature babies in England (BEADI) study protocol [ISRCTN89683698]. AB - BACKGROUND: Gaps between research knowledge and practice have been consistently reported. Traditional ways of communicating information have limited impact on practice changes. Strategies to disseminate information need to be more interactive and based on techniques reported in systematic reviews of implementation of changes. There is a need for clarification as to which dissemination strategies work best to translate evidence into practice in neonatal units across England. The objective of this trial is to assess whether an innovative active strategy for the dissemination of neonatal research findings, recommendations, and national neonatal guidelines is more likely to lead to changes in policy and practice than the traditional (more passive) forms of dissemination in England. METHODS/DESIGN: Cluster randomised controlled trial of all neonatal units in England (randomised by hospital, n = 182 and stratified by neonatal regional networks and neonatal units level of care) to assess the relative effectiveness of active dissemination strategies on changes in local policies and practices. Participants will be mainly consultant lead clinicians in each unit. The intervention will be multifaceted using: audit and feedback; educational meetings for local staff (evidence-based lectures on selected topics, interactive workshop to examine current practice and draw up plans for change); and quality improvement and organisational changes methods. Policies and practice outcomes for the babies involved will be collected before and after the intervention. Outcomes will assess all premature babies born in England during a three month period for timing of surfactant administration at birth, temperature control at birth, and resuscitation team (qualification and numbers) present at birth. PMID- 17922902 TI - A patient with bilateral pheochromocytoma as part of a Von Hippel-Lindau (VHL) syndrome type 2C. AB - BACKGROUND: Von Hippel-Lindau (VHL) disease is an autosomal dominant inherited disease. It is relatively recent that type 2C was identified as a separate group solely presenting with pheochromocytomas. As an illustration, an interesting case is presented of a pregnant woman with refractory hypertension. It proved to be the first manifestation of bilateral pheochromocytomas. The family history may indicate the diagnosis, but only identification of a germ line mutation in the DNA of a patient will confirm carriership. CASE PRESENTATION: A 27 year pregnant patient with intra uterine growth retardation presented with hypertension and pre eclampsia. Magnetic resonance imaging revealed bilateral adrenal pheochromocytoma. She underwent laparoscopic adrenelectomy and a missense mutation (Gly93Ser) in exon 1 of the VHL gene on chromosome 3 (p25 - p26) was shown in the patient, her father and her daughter confirming the diagnosis of VHL. CONCLUSION: In almost all VHL families molecular genetic analysis of DNA will demonstrate an inherited mutation. Because of the involvement in several organs, periodic clinical evaluation should take place in a well coordinated, multidisciplinary setting. VHL disease can be classified into several subtypes. VHL type 2C patients present with pheochromocytomas without evidence of haemangioblastomas in the central nervous system and/or retina and a low risk of renal cell carcinoma. Therefore, in such families, periodic clinical screening can be focussed on pheochromocytomas. PMID- 17922903 TI - Structural disorder promotes assembly of protein complexes. AB - BACKGROUND: The idea that the assembly of protein complexes is linked with protein disorder has been inferred from a few large complexes, such as the viral capsid or bacterial flagellar system, only. The relationship, which suggests that larger complexes have more disorder, has never been systematically tested. The recent high-throughput analyses of protein-protein interactions and protein complexes in the cell generated data that enable to address this issue by bioinformatic means. RESULTS: In this work we predicted structural disorder for both E. coli and S. cerevisiae, and correlated it with the size of complexes. Using IUPred to predict the disorder for each complex, we found a statistically significant correlation between disorder and the number of proteins assembled into complexes. The distribution of disorder has a median value of 10% in yeast for complexes of 2-4 components (6% in E. coli), but 18% for complexes in the size range of 11-100 proteins (12% in E. coli). The level of disorder as assessed for regions longer than 30 consecutive disordered residues shows an even stronger division between small and large complexes (median values about 4% for complexes of 2-4 components, but 12% for complexes of 11-100 components in yeast). The predicted correlation is also supported by experimental evidence, by observing the structural disorder in protein components of complexes that can be found in the Protein Data Bank (median values 1. 5% for complexes of 2-4 components, and 9.6% for complexes of 11-100 components in yeast). Further analysis shows that this correlation is not directly linked with the increased disorder in hub proteins, but reflects a genuine systemic property of the proteins that make up the complexes. CONCLUSION: Overall, it is suggested and discussed that the assembly of protein-protein complexes is enabled and probably promoted by protein disorder. PMID- 17922904 TI - Suicide in deaf populations: a literature review. AB - BACKGROUND: Studies have found that deaf individuals have higher rates of psychiatric disorder than those who are hearing, while at the same time encountering difficulties in accessing mental health services. These factors might increase the risk of suicide. However, the burden of suicidal behaviour in deaf people is currently unknown. The aim of the present review was to provide a summary of literature on suicidal behaviour with specific reference to deaf individuals. The objectives of the review were to establish the incidence and prevalence of suicidal behaviour in deaf populations; describe risk factors for suicidal behaviour in deaf populations; describe approaches to intervention and suicide prevention that have been used in deaf populations. METHODS: A number of electronic databases (e.g. Medline, PsycINFO, CINAHL, EMBASE, Dissertation Abstracts International, Web of Science, ComDisDome, ASSIA, Education Sage Full Text, Google Scholar, and the grey literature databases FADE and SIGLE) were explored using a combination of key words and medical subject headings as search terms. Reference lists of papers were also searched. The Science and Social Sciences Citation Index electronic databases were used to identify studies that had cited key papers. We also contacted experts and organisations with an interest in the field. RESULTS: Very few studies focussed specifically on suicide in deaf populations. Those studies that were included (n = 13) generally involved small and unrepresentative samples. There were limited data on the rate of suicidal behaviour in deaf people. One study reported evidence of hearing impairment in 0.2% of all suicide deaths. Another found that individuals with tinnitus seen in specialist clinics had an elevated rate of suicide compared to the general population. The rates of attempted suicide in deaf school and college students during the previous year ranged from 1.7% to 18%, with lifetime rates as high as 30%. Little evidence was found to suggest that risk factors for suicide in deaf people differed systematically from those in the general population. However, studies did report higher levels of depression and higher levels of perceived risk among deaf individuals than hearing control groups. No firm evidence was found regarding the effectiveness of suicide prevention strategies in deaf people, but suggested strategies include developing specific screening tools, training clinical staff, promoting deaf awareness, increasing the availability of specialist mental health services for deaf people. CONCLUSION: There is a significant gap in our understanding of suicide in deaf populations. Clinicians should be aware of the possible association between suicide and deafness. Specialist mental health services should be readily accessible to deaf individuals and specific preventative strategies may be of benefit. However, further research using a variety of study designs is needed to increase our understanding of this issue. PMID- 17922905 TI - Postprandial ghrelin suppression is exaggerated following major surgery; implications for nutritional recovery. AB - Meeting patients' nutritional requirements and preventing malnutrition is a challenge following major surgical procedures. The role of ghrelin in nutritional recovery after non-gastrointestinal major surgery is unknown. We used coronary artery bypass grafting (CABG) as an example of anticipated good recovery post major surgery.Seventeen patients undergoing CABG (mean +/- SEM: 70.1 +/- 2.2 yrs, BMI 29.1 +/- 1.4 kg/m2, 15 male) underwent fasting and postprandial (45 mins after standard test breakfast) blood sampling pre-operatively (day 0), post operatively (day 6) and at follow-up (day 40). Changes in food intake, biochemical and anthropometric markers of nutritional status were recorded. A comparison was made to 17 matched healthy controls (70.6 +/- 2.3 yrs, BMI 28.4 +/ 1.3 kg/m2).We observed significantly increased post-operative and follow-up fasting ghrelin concentrations compared with pre-operatively (pre-op. 402 +/- 42 pmol/L vs post-op. 642 +/- 97 pmol/L vs follow-up 603 +/- 94 pmol/L) (ANOVA p < 0.05). Significantly exaggerated postprandial suppression of ghrelin was seen postoperatively and continued until follow-up (Delta pre-op. 10 +/- 51 pmol/L vs Delta post-op. -152 +/- 43 pmol/L vs Delta follow-up -159 +/- 65 pmol/L, p < 0.05). This was associated with a 50% reduction in food intake {post-op. 4.5 +/- 0.5 MJ/D (1076 +/- 120 kcal/D) compared with estimated requirements 9.9 +/- 0.5 MJ/D (2366 +/- 120 kcal/D)}, leading to a 4% weight loss and a 5% reduction in muscle arm circumference loss over length of follow up.Our data support the hypothesis that prolonged changes in fasting and postprandial plasma ghrelin concentrations are associated with impaired nutritional recovery after CABG. These findings reinforce the need to investigate ghrelin in other patients groups undergoing major surgery. PMID- 17922906 TI - Intrinsic genetic characteristics determine tumor-modifying capacity of fibroblasts: matrix metalloproteinase-3 5A/5A genotype enhances breast cancer cell invasion. AB - BACKGROUND: Stromal fibroblasts can contribute to tumor invasion through the release of matrix metalloproteinases (MMPs). Population studies have suggested that single nucleotide polymorphisms (SNPs) in MMP genes influence levels of expression and may be associated with breast cancer risk and with disease progression. This study directly examined the impact of MMP SNP genotype on the ability of host fibroblasts to promote tumor cell invasion. METHODS: Primary breast fibroblasts were isolated from patients with (n = 13) or without (n = 19) breast cancer, and their ability to promote breast cancer cell invasion was measured in in vitro invasion assays. Fibroblast invasion-promoting capacity (IPC) was analyzed in relation to donor type (tumor or non-tumor patient), MMP-1, MMP-3, and MMP-9 SNP genotype and MMP activity using independent samples t test and analysis of variance. All statistical tests were two-sided. RESULTS: Tumor derived fibroblasts promoted higher levels of invasion than normal fibroblasts (p = 0.041). When IPC was related to genotype, higher levels of IPC were generated by tumor fibroblasts with the high-expressing MMP-3 5A/5A genotype compared with the 5A/6A and 6A/6A genotypes (p = 0.05 and 0.07, respectively), and this was associated with enhanced MMP-3 release. The functional importance of MMP-3 was demonstrated by enhanced invasion in the presence of recombinant MMP-3, whereas reduction occurred in the presence of a specific MMP-3 inhibitor. An inverse relationship was demonstrated between fibroblast IPC and the high-expressing MMP 1 genotype (p = 0.031), but no relationship was seen with MMP-9 SNP status. In contrast, normal fibroblasts showed no variation in IPC in relation to MMP genotype, with MMP-3 5A/5A fibroblasts exhibiting significantly lower levels of IPC than their tumor-derived counterparts (p = 0.04). CONCLUSION: This study has shown that tumor-derived fibroblasts exhibit higher levels of IPC than normal fibroblasts and that the MMP-3 5A/5A genotype contributes to this through enhanced MMP-3 release. Despite a high-expressing genotype, normal fibroblasts do not exhibit higher IPC or enhanced MMP release. This suggests that more complex changes occur in tumor-derived fibroblasts, enabling full expression of the MMP SNP genotype and these possibly are epigenetic in nature. The results do suggest that, in women with breast cancer, a high-expressing MMP-3 genotype may promote tumor progression more effectively. PMID- 17922907 TI - Elevated extracellular matrix production and degradation upon bone morphogenetic protein-2 (BMP-2) stimulation point toward a role for BMP-2 in cartilage repair and remodeling. AB - Bone morphogenetic protein-2 (BMP-2) has been proposed as a tool for cartilage repair and as a stimulant of chondrogenesis. In healthy cartilage, BMP-2 is hardly present, whereas it is highly expressed during osteoarthritis. To assess its function in cartilage, BMP-2 was overexpressed in healthy murine knee joints and the effects on proteoglycan (PG) synthesis and degradation were evaluated. Moreover, the contribution of BMP in repairing damage induced by interleukin-1 (IL-1) was investigated. Ad-BMP-2 was injected intra-articularly into murine knee joints, which were isolated 3, 7, and 21 days after injection for histology, immunohistochemistry, and autoradiography. In addition, patellar and tibial cartilage was isolated for RNA isolation or measurement of PG synthesis by means of 35SO4(2-) incorporation. To investigate the role for BMP-2 in cartilage repair, cartilage damage was induced by intra-articular injection of IL-1. After 2 days, Ad-BMP-2, Ad-BMP-2 + Ad-gremlin, Ad-gremlin, or a control virus was injected. Whole knee joints were isolated for histology at day 4 or patellae were isolated to measure 35SO4(2-) incorporation. BMP-2 stimulated PG synthesis in patellar cartilage on all days and in tibial cartilage on day 21. Aggrecan mRNA expression had increased on all days in patellar cartilage, with the highest increase on day 7. Collagen type II expression showed a similar expression pattern. In tibial cartilage, collagen type II and aggrecan mRNA expression had increased on days 7 and 21. BMP-2 overexpression also induced increased aggrecan degradation in cartilage. VDIPEN staining (indicating matrix metalloproteinase activity) was elevated on day 3 in tibial cartilage and on days 3 and 7 in patellar cartilage, but no longer was by day 21. Increased NITEGE staining (indicating aggrecanase activity) was found on days 7 and 21. In IL-1-damaged patellar cartilage, BMP-2 boosted PG synthesis. Blocking of BMP activity resulted in a decreased PG synthesis compared with IL-1 alone. This decreased PG synthesis was associated with PG depletion in the cartilage. These data show that BMP-2 boosts matrix turnover in intact and IL-damaged cartilage. Moreover, BMP contributes to the intrinsic repair capacity of damaged cartilage. Increased matrix turnover might be functional in replacing matrix molecules in the repair of a damaged cartilage matrix. PMID- 17922908 TI - Aorto-ventricular tunnel. AB - Aorto-ventricular tunnel is a congenital, extracardiac channel which connects the ascending aorta above the sinutubular junction to the cavity of the left, or (less commonly) right ventricle. The exact incidence is unknown, estimates ranging from 0.5% of fetal cardiac malformations to less than 0.1% of congenitally malformed hearts in clinico-pathological series. Approximately 130 cases have been reported in the literature, about twice as many cases in males as in females. Associated defects, usually involving the proximal coronary arteries, or the aortic or pulmonary valves, are present in nearly half the cases. Occasional patients present with an asymptomatic heart murmur and cardiac enlargement, but most suffer heart failure in the first year of life. The etiology of aorto-ventricular tunnel is uncertain. It appears to result from a combination of maldevelopment of the cushions which give rise to the pulmonary and aortic roots, and abnormal separation of these structures. Echocardiography is the diagnostic investigation of choice. Antenatal diagnosis by fetal echocardiography is reliable after 18 weeks gestation. Aorto-ventricular tunnel must be distinguished from other lesions which cause rapid run-off of blood from the aorta and produce cardiac failure. Optimal management of symptomatic aorto ventricular tunnel consists of diagnosis by echocardiography, complimented with cardiac catheterization as needed to elucidate coronary arterial origins or associated defects, and prompt surgical repair. Observation of the exceedingly rare, asymptomatic patient with a small tunnel may be justified by occasional spontaneous closure. All patients require life-long follow-up for recurrence of the tunnel, aortic valve incompetence, left ventricular function, and aneurysmal enlargement of the ascending aorta. PMID- 17922909 TI - Genetic knockout and pharmacologic inhibition of neuronal nitric oxide synthase attenuate nerve injury-induced mechanical hypersensitivity in mice. AB - Neuronal nitric oxide synthase (nNOS) is a key enzyme for nitric oxide production in neuronal tissues and contributes to the spinal central sensitization in inflammatory pain. However, the role of nNOS in neuropathic pain remains unclear. The present study combined a genetic strategy with a pharmacologic approach to examine the effects of genetic knockout and pharmacologic inhibition of nNOS on neuropathic pain induced by unilateral fifth lumbar spinal nerve injury in mice. In contrast to wildtype mice, nNOS knockout mice failed to display nerve injury induced mechanical hypersensitivity. Furthermore, either intraperitoneal (100 mg/kg) or intrathecal (30 microg/5 microl) administration of L-NG-nitro-arginine methyl ester, a nonspecific NOS inhibitor, significantly reversed nerve injury induced mechanical hypersensitivity on day 7 post-nerve injury in wildtype mice. Intrathecal injection of 7-nitroindazole (8.15 microg/5 microl), a selective nNOS inhibitor, also dramatically attenuated nerve injury-induced mechanical hypersensitivity. Western blot analysis showed that the expression of nNOS protein was significantly increased in ipsilateral L5 dorsal root ganglion but not in ipsilateral L5 lumbar spinal cord on day 7 post-nerve injury. The expression of inducible NOS and endothelial NOS proteins was not markedly altered after nerve injury in either the dorsal root ganglion or spinal cord. Our findings suggest that nNOS, especially in the dorsal root ganglion, may participate in the development and/or maintenance of mechanical hypersensitivity after nerve injury. PMID- 17922910 TI - Molecular epidemiology of Hepatitis B virus genotypes in Pakistan. AB - BACKGROUND: Eight genotypes of Hepatitis B virus designated A-H, have been known but in Pakistan, no such data is available on the prevalent HBV genotypes. Therefore, the subject study was conducted to determine HBV genotypes in the indigenous Pakistani population. METHODS: A total of 690 individuals were enrolled for HBV screening with EIA and nested PCR. Positive samples were further analyzed to determine HBV genotypes (A-F) by multiplex-PCR using type specific primers. RESULTS: 110 (15.94%) individuals were positive for HBV, including 64% males and 36% females. Out of these, 66 samples (65.34%) were classified into genotype D, 27 (26.73%) were of genotype B while 5(4.95%) had genotype A. In 3 (2.98%) samples, multiple genotypes were detected (genotype A+B; 2(1.99%) and genotypes B+D; 1(0.99%). Nine (8.18%) samples remained untyable. CONCLUSION: In Asia, genotypes B and C are the most prevalent but our study reveals that genotype D is predominant and HBV infection constitutes a significant health problem in Pakistan. PMID- 17922912 TI - Immune restoration syndrome with disseminated Penicillium marneffei and cytomegalovirus co-infections in an AIDS patient. AB - BACKGROUND: Penicillium marneffei is a dimorphic fungus, endemic in South-east Asia. The fungus causes severe disease in immunocompromised patients such as AIDS. However, no case of immune restoration disease of Penicillium marneffei is reported in literature from a non-endemic area. CASE PRESENTATION: We report the first case of Penicillium marneffei and Cytomegalovirus infection manifesting as a result of immune restoration one month after initiating HAART. This severely immunocompromised patient had presented with multiple lymphadenopathy, massive hepatosplenomegaly, visual impairment and mild icterus, but no skin lesions. Penicillium marneffei was isolated from lymph node fine-needle aspirates and blood cultures. CONCLUSION: In order to diagnose such rare cases, the clinicians, histopathologists and microbiologists alike need to maintain a strong index of suspicion for making initial diagnosis as well as for suspecting immune reconstitution syndrome (IRS) with Penicillium marneffei. PMID- 17922911 TI - MicroRNA expression profiling of human breast cancer identifies new markers of tumor subtype. AB - BACKGROUND: MicroRNAs (miRNAs), a class of short non-coding RNAs found in many plants and animals, often act post-transcriptionally to inhibit gene expression. RESULTS: Here we report the analysis of miRNA expression in 93 primary human breast tumors, using a bead-based flow cytometric miRNA expression profiling method. Of 309 human miRNAs assayed, we identify 133 miRNAs expressed in human breast and breast tumors. We used mRNA expression profiling to classify the breast tumors as luminal A, luminal B, basal-like, HER2+ and normal-like. A number of miRNAs are differentially expressed between these molecular tumor subtypes and individual miRNAs are associated with clinicopathological factors. Furthermore, we find that miRNAs could classify basal versus luminal tumor subtypes in an independent data set. In some cases, changes in miRNA expression correlate with genomic loss or gain; in others, changes in miRNA expression are likely due to changes in primary transcription and or miRNA biogenesis. Finally, the expression of DICER1 and AGO2 is correlated with tumor subtype and may explain some of the changes in miRNA expression observed. CONCLUSION: This study represents the first integrated analysis of miRNA expression, mRNA expression and genomic changes in human breast cancer and may serve as a basis for functional studies of the role of miRNAs in the etiology of breast cancer. Furthermore, we demonstrate that bead-based flow cytometric miRNA expression profiling might be a suitable platform to classify breast cancer into prognostic molecular subtypes. PMID- 17922913 TI - The impact of infliximab treatment on quality of life in patients with inflammatory rheumatic diseases. AB - In this study, we compare the health-related quality of life (HRQoL) of patients with moderate-to-severe rheumatoid arthritis (RA), psoriatic arthritis (PsA), and ankylosing spondylitis (AS), and study the effect of treatment with infliximab on the HRQoL of patients with these diseases. Short Form Health Survey-36 (SF-36) data from the placebo-controlled phases of 4 studies of infliximab in patients with inflammatory rheumatic diseases (n = 1990) were evaluated. Data came from the Anti-TNF Trial in Rheumatoid Arthritis with Concomitant Therapy (ATTRACT) (n = 428), the Safety Trial for Rheumatoid Arthritis with REMICADE Therapy (START) (n = 1083), the Ankylosing Spondylitis Study for the Evaluation of Recombinant Infliximab Therapy (ASSERT) (n = 279), and the Infliximab Multinational Psoriatic Arthritis Clinical Trial II (IMPACT II) (n = 200). SF-36 assessments were made at weeks 0, 10, 30, and 54 in ATTRACT, weeks 0, 6, and 22 in START, weeks 0, 12, and 24 in ASSERT, and weeks 0 and 14 in IMPACT II. All patient populations had significantly impaired physical aspects of HRQoL at baseline relative to the general population of the United States, and the magnitude of impairment was similar across the diseases. Mean baseline physical component summary scores were 29 in the RA cohort, 32 in the PsA cohort, and 29 in the AS cohort. In all 3 diseases, patients who received infliximab showed significant improvement in physical component summary scores compared with those who received placebo. The magnitude of the difference of improvement (effect size, 95%CI) between infliximab and placebo groups was similar in the AS (10.1, 9.2-11.0), PsA (8.6, 7.8-9.4), and RA (10.1, 9.2-11.0) cohorts. Patients with RA and those with PsA treated with infliximab also showed greater improvement in the mental component summary score than those in the placebo group with an effect size of 4.6 (4.2 5.1) in RA and 2.7 (2.4-3.1) in PsA. Patients in large randomized controlled studies of infliximab in RA, PsA, and AS had similar impairment in physical aspects of HRQoL at baseline and showed significantly greater improvement in HRQoL after treatment with infliximab. PMID- 17922914 TI - Respiratory function and bronchial responsiveness among industrial workers exposed to different classes of occupational agents: a study from Algeria. AB - Occupational exposures play a role in the onset of several chronic airway diseases. We investigated, in a cross-sectional study, lung function parameters and bronchial hyper-responsiveness to histamine in workers exposed to different airborne compounds.The study group totalled 546 male subjects of whom 114 were exposed to welding fumes, 106 to solvents, 107 to mineral dust, 97 to organic dust and 123 without known exposure to airway irritants. A questionnaire was administered and spirometry and bronchial responsiveness to histamine were assessed by one observer, in the morning before work to prevent effects of acute exposure.The mean (SD) age of the participants was 39.3 (7.8) years, with a mean duration of employment of 13.8 (6.6) years. Both before and after adjustment for smoking status, forced expiratory volume in 1 second (FEV1, expressed as % predicted) was lower in welders -4.0% (95% confidence interval [CI], -6.3 to 1.8; p = 0.01) and workers exposed to solvents -5.6% (CI: -7.9 to -3.3; p = 0.0009) than in control subjects. Furthermore, solvent workers had an odds ratio of 3.43 (95% CI: 1.09-11.6; p = 0.037) for bronchial hyperresponsiveness compared with the reference group.The higher prevalence of bronchial hyperresponsiveness in solvent workers adds to the growing body of evidence of adverse respiratory effects of occupational solvent exposure. These results point to the necessity of preventive measures in solvent workers to avoid these adverse respiratory effects. PMID- 17922915 TI - Percutaneous tracheostomy in patients with severe liver disease and a high incidence of refractory coagulopathy: a prospective trial. AB - BACKGROUND: To assess the safety of percutaneous dilational tracheostomy (PDT) performed by experienced operators in critically ill patients with liver disease and coagulopathy. METHODS: Prospective cohort study in a ten bed specialist liver intensive care unit of a tertiary university teaching hospital. Sixty consecutive patients in need of tracheostomy insertion. Patients were categorized as having refractory coagulopathy if their platelet count was < or = 50 x 10(9) cells/L or the INR > 1.5 on the day of and the subsequent 72 hours following PDT despite clotting support. RESULTS: Twenty five patients fulfilled the definition criteria of refractory coagulopathy. There was no significant difference in the number of adverse incidents between groups. Only 1 patient in the coagulopathy group had a severe bleeding complication, however this did not require open surgical intervention. The rate of clinically relevant early complications in all patients was not higher than expected (n = 7, 12%). Resource utilisation was higher for patients with coagulopathy, who received significantly more platelet transfusions over the 3 day period (80 vs 49 units, p = 0.009) and demonstrated a trend towards increased fresh frozen plasma requirements (p = 0.059). The number of patients requiring platelet transfusion was higher in the coagulopathy group (21/25 versus 20/35 p = 0.029). Hospital survival did not differ between groups. CONCLUSION: PDT is safe and not contraindicated in patients with severe liver disease and refractory coagulopathy. PMID- 17922916 TI - Impact of home-based management of malaria on health outcomes in Africa: a systematic review of the evidence. AB - BACKGROUND: Home-based management of malaria (HMM) is promoted as a major strategy to improve prompt delivery of effective malaria treatment in Africa. HMM involves presumptively treating febrile children with pre-packaged antimalarial drugs distributed by members of the community. HMM has been implemented in several African countries, and artemisinin-based combination therapies (ACTs) will likely be introduced into these programmes on a wide scale. CASE PRESENTATIONS: The published literature was searched for studies that evaluated the health impact of community- and home-based treatment for malaria in Africa. Criteria for inclusion were: 1) the intervention consisted of antimalarial treatment administered presumptively for febrile illness; 2) the treatment was administered by local community members who had no formal education in health care; 3) measured outcomes included specific health indicators such as malaria morbidity (incidence, severity, parasite rates) and/or mortality; and 4) the study was conducted in Africa. Of 1,069 potentially relevant publications identified, only six studies, carried out over 18 years, were identified as meeting inclusion criteria. Heterogeneity of the evaluations, including variability in study design, precluded meta-analysis. DISCUSSION AND EVALUATION: All trials evaluated presumptive treatment with chloroquine and were conducted in rural areas, and most were done in settings with seasonal malaria transmission. Conclusions regarding the impact of HMM on morbidity and mortality endpoints were mixed. Two studies showed no health impact, while another showed a decrease in malaria prevalence and incidence, but no impact on mortality. One study in Burkina Faso suggested that HMM decreased the proportion of severe malaria cases, while another study from the same country showed a decrease in the risk of progression to severe malaria. Of the four studies with mortality endpoints only one from Ethiopia showed a positive impact, with a reduction in the under-5 mortality rate of 40.6% (95% CI 29.2 - 50.6). CONCLUSION: Currently the evidence base for HMM in Africa, particularly regarding use of ACTs, is narrow and priorities for further research are discussed. To optimize treatment and maximize health benefits, drug regimens and delivery strategies in HMM programmes may need to be tailored to local conditions. Additional research could help guide programme development, policy decision-making, and implementation. PMID- 17922917 TI - Erwinia carotovora elicitors and Botrytis cinerea activate defense responses in Physcomitrella patens. AB - BACKGROUND: Vascular plants respond to pathogens by activating a diverse array of defense mechanisms. Studies with these plants have provided a wealth of information on pathogen recognition, signal transduction and the activation of defense responses. However, very little is known about the infection and defense responses of the bryophyte, Physcomitrella patens, to well-studied phytopathogens. The purpose of this study was to determine: i) whether two representative broad host range pathogens, Erwinia carotovora ssp. carotovora (E.c. carotovora) and Botrytis cinerea (B. cinerea), could infect Physcomitrella, and ii) whether B. cinerea, elicitors of a harpin (HrpN) producing E.c. carotovora strain (SCC1) or a HrpN-negative strain (SCC3193), could cause disease symptoms and induce defense responses in Physcomitrella. RESULTS: B. cinerea and E.c. carotovora were found to readily infect Physcomitrella gametophytic tissues and cause disease symptoms. Treatments with B. cinerea spores or cell-free culture filtrates from E.c. carotovoraSCC1 (CF(SCC1)), resulted in disease development with severe maceration of Physcomitrella tissues, while CF(SCC3193) produced only mild maceration. Although increased cell death was observed with either the CFs or B. cinerea, the occurrence of cytoplasmic shrinkage was only visible in Evans blue stained protonemal cells treated with CF(SCC1) or inoculated with B. cinerea. Most cells showing cytoplasmic shrinkage accumulated autofluorescent compounds and brown chloroplasts were evident in a high proportion of these cells. CF treatments and B. cinerea inoculation induced the expression of the defense-related genes: PR-1, PAL, CHS and LOX. CONCLUSION: B. cinerea and E.c. carotovora elicitors induce a defense response in Physcomitrella, as evidenced by enhanced expression of conserved plant defense related genes. Since cytoplasmic shrinkage is the most common morphological change observed in plant PCD, and that harpins and B. cinerea induce this type of cell death in vascular plants, our results suggest that E.c. carotovora CFSCC1 containing HrpN and B. cinerea could also induce this type of cell death in Physcomitrella. Our studies thus establish Physcomitrella as an experimental host for investigation of plant-pathogen interactions and B. cinerea and elicitors of E.c. carotovora as promising tools for understanding the mechanisms involved in defense responses and in pathogen-mediated cell death in this simple land plant. PMID- 17922918 TI - Environmental correlates of physical activity in multiple sclerosis: a cross sectional study. AB - BACKGROUND: Multiple sclerosis (MS) is a chronic neurological disease that is associated with physical inactivity. Understanding the factors that correlate with physical activity is important for developing effective physical activity promotion programs for this population. Thus, we conducted a cross-sectional study that examined the association between features of the built environment with self-reported and objectively measured physical activity behaviour in adults with MS. METHODS: Participants with MS (n = 196) were sent a questionnaire packet that included self-report measures of the built environment and physical activity and a pedometer in the mail and were instructed to complete the questionnaires and wear the device for seven days. Participants returned the completed questionnaires in a pre-stamped, pre-addressed envelope. Bivariate correlation analysis was conducted for examining associations between items on the environmental questionnaire with the two measures of physical activity. Stepwise regression analysis was conducted for determining the independent contributions of the significant environmental correlates for explaining variation in physical activity. RESULTS: Correlational analysis indicated that presence of shops, stores, markets or other places within walking distance (r = .20; rho = .18), presence of a transit stop within walking distance (r = .20; rho = .16), and accessibility of free or low-cost recreation facilities (r = .16; rho = .15) were related to pedometer, but not self-reported, measured physical activity. Regression analysis indicated that the presence of a transit stop within walking distance independently explained 4% of variance in pedometer measured physical activity. CONCLUSION: Physical activity is an important behaviour to promote among individuals with MS. This study indicated that aspects of the built environment are related to this health promoting behaviour among those with MS. Further research should focus on the longitudinal relationships among aspects of the environment with physical activity so as to provide strong background for developing effective promotion programs for people with MS. PMID- 17922919 TI - Circulating surfactant protein D as a potential lung-specific biomarker of health outcomes in COPD: a pilot study. AB - BACKGROUND: There is a paucity of surrogate lung-specific biological markers that can be used to track disease progression and predict clinical outcomes in chronic obstructive pulmonary disease (COPD). The principal aim of this pilot study was to determine whether circulating surfactant protein D (SPD) or Clara Cell protein 16 (CC16) levels are associated with lung function or health status in patients with severe COPD. METHODS: We studied 23 patients with advanced COPD. Lung function measurements, Chronic Respiratory Disease Questionnaire (CRQ) scores, and serum levels of SPD, CC16, and C-reactive protein (CRP) were determined at baseline and at 3 months. RESULTS: At baseline, FEV(1) was inversely associated with serum SPD levels (P = 0.045) but not with CC16 (P = 0.675) or CRP levels (P = 0.549). Over a 3 month period, changes in SPD levels correlated significantly with changes in CRQ scores (adjusted P = 0.008) such that patients who had the largest declines in serum SPD levels experienced the largest gains in health status. The association was particularly notable between circulating SPD level and the dyspnea domain of the CRQ score (P = 0.018). Changes in CC16 or CRP levels did not correlate with changes in CRQ scores. CONCLUSION: Changes in serum SPD levels tracked well with changes in health status over a 3 month period in patients with severe COPD. These data suggest that circulating SPD levels may be useful biomarkers to track health outcomes of COPD patients. PMID- 17922920 TI - Chromogenic in situ hybridisation for the assessment of HER2 status in breast cancer: an international validation ring study. AB - INTRODUCTION: Before any new methodology can be introduced into the routine diagnostic setting it must be technically validated against the established standards. To this end, a ring study involving five international pathology laboratories was initiated to validate chromogenic in situ hybridisation (CISH) against fluorescence in situ hybridisation (FISH) and immunohistochemistry (IHC) as a test for assessing human epidermal growth factor receptor 2 (HER2) status in breast cancer. METHODS: Each laboratory performed CISH, FISH and IHC on its own samples. Unstained sections from each case were also sent to another participating laboratory for blinded retesting by CISH ('outside CISH'). RESULTS: A total of 211 invasive breast carcinoma cases were tested. In 76 cases with high amplification (HER2/CEP17 ratio >4.0) by FISH, 73 cases (96%) scored positive (scores >or= 6) by 'outside CISH'. For FISH-negative cases (HER2/CEP17 ratio <2.0), 94 of 100 cases (94%) had CISH scores indicating no amplification (score or= 6. CISH intra-laboratory concordance with IHC was 92% for IHC-negative cases (IHC 0/1+) and 91% for IHC 3+ cases. Among IHC 2+ cases, CISH was 100% concordant with samples showing high amplification by FISH, and 94% concordant with FISH-negative samples. CONCLUSION: These results show that CISH inter- and intra-laboratory concordance to FISH and IHC is very high, even in equivocal IHC 2+ cases. Therefore, we conclude that CISH is a methodology that is a viable alternative to FISH in the HER2 testing algorithm. PMID- 17922921 TI - An unusual case of gout in the wrist: the importance of monitoring medication dosage and interaction. A case report. AB - BACKGROUND: Gouty arthritis of the wrist is uncommon although gout itself is the most common inflammatory arthritis in older patients. Some known risk factors for the development of gout include trauma, alcohol use, obesity, hyperuricaemia, hypertension and diabetes mellitus. As well, certain medications have been shown to promote the development of gout. These include thiazide diuretics, low dose salicylates and cyclosporine. We present a case of gouty wrist pain possibly precipitated by a medication dosage increase as well as medication interactions. CASE PRESENTATION: A 77 year old male presented with right wrist pain. Redness and swelling was present at the dorsal aspect of his wrist and range of motion was full with pain at end range upon examination. One week prior, his anti hypertensive medication dosage had been increased. The patient's situation continued to worsen. Radiographic examination revealed changes consistent with gouty arthritis. CONCLUSION: It is important for clinicians treating joint conditions to be aware of patients' comorbidities, medication usage and changes in dosages. Education of patients with gout is of prime importance. Clinicians should educate patients that gout may occur at any joint in the body not only the lower limb. Patients should be aware of the signs and symptoms of an acute gouty attack and be made aware that changes in certain medication dosages may precipitate an attack. Awareness of radiographic changes associated with gout is still of importance although these changes are not seen as frequently as they have been in the past due to better control of the disease. PMID- 17922922 TI - Effects of visually simulated roll motion on vection and postural stabilization. AB - BACKGROUND: Visual motion often provokes vection (the induced perception of self motion) and postural movement. Postural movement is known to increase during vection, suggesting the same visual motion signal underlies vection and postural control. However, self-motion does not need to be consciously perceived to influence postural control. Therefore, visual motion itself may affect postural control mechanisms. The purpose of the present study was to investigate the effects of visual motion and vection on postural movements during and after exposure to a visual stimulus motion. METHODS: Eighteen observers completed four experimental conditions, the order of which was counterbalanced across observers. Conditions corresponded to the four possible combinations of rotation direction of the visually simulated roll motion stimulus and the two different visual stimulus patterns. The velocity of the roll motion was held constant in all conditions at 60 deg/s. Observers assumed the standard Romberg stance, and postural movements were measured using a force platform and a head position sensor affixed to a helmet they wore. Observers pressed a button when they perceived vection. Postural responses and psychophysical parameters related to vection were analyzed. RESULTS: During exposure to the moving stimulus, body sway and head position of all observers moved in the same direction as the stimulus. Moreover, they deviated more during vection perception than no-vection perception, and during no-vection-perception than no-visual-stimulus-motion. The postural movements also fluctuated more during vection-perception than no-vection perception, and during no-vection-perception than no-visual-stimulus-motion, both in the left/right and anterior/posterior directions. There was no clear habituation for vection and posture, and no effect of stimulus type. CONCLUSION: Our results suggested that visual stimulus motion itself affects postural control, and supported the idea that the same visual motion signal is used for vection and postural control. We speculated that the mechanisms underlying the processing of visual motion signals for postural control and vection perception operate using different thresholds, and that a frame of reference for body orientation perception changed along with vection perception induced further increment of postural sway. PMID- 17922923 TI - High prevalence of lack of knowledge of symptoms of acute myocardial infarction in Pakistan and its contribution to delayed presentation to the hospital. AB - BACKGROUND: We conducted an observational study to determine the delay in presentation to hospital, and its associates among patients experiencing first Acute Myocardial Infarction (AMI) in Karachi, Pakistan. METHODS: A hospital based cross-sectional study was conducted at National Institute of Cardiovascular Disease (NICVD) in Karachi. A structured questionnaire was used to collect data. The primary outcome was delay in presentation, defined as a time interval of six or more hours from the onset of symptoms to presentation to hospital. Logistic regression analysis was performed to determine the factors associated with prehospital delay. RESULTS: A total of 720 subjects were interviewed; 22% were females. The mean age (SD) of the subjects was 54 (+/- 12) years. The mean (SE) and median (IQR) time to presentation was 12.3 (1.7) hours and 3.04 (6.0) hours respectively. About 34% of the subjects presented late. Lack of knowledge of any of the symptoms of heart attack (odds ratio (95% CI)) (1.82 (1.10, 2.99)), and mild chest pain (10.05 (6.50, 15.54)) were independently associated with prehospital delay. CONCLUSION: Over one-third of patients with AMI in Pakistan present late to the hospital. Lack of knowledge of symptoms of heart attack, and low severity of chest pain were the main predictors of prehospital delay. Strategies to reduce delayed presentation in this population must focus on education about symptoms of heart attack. PMID- 17922924 TI - Nuclear localization of beta-catenin involved in precancerous change in oral leukoplakia. AB - BACKGROUND: Oral leukoplakia is a precancerous change developed in the oral mucosa, and the mechanism that oral leukoplakia becomes malignant through atypical epithelium is not known. Here we compared the beta-catenin expression detected by immunohistochemical staining in the normal oral epithelium and in the oral leukoplakia with or without dysplasia. RESULTS: The normal oral epithelium showed beta-catenin expression only in the cell membrane, but not in the nuclei. In the oral leukoplakia without dysplasia, 7 out of 17 samples (41%) showed beta catenin expression in the cell membrane, and 5 samples (29%) showed expression in the nuclei. In the oral leukoplakia with dysplasia, nuclear expression of beta catenin was shown in 11 out of 12 samples (92%). Incidence of nuclear beta catenin expression was significantly different between dysplasia and normal oral epithelium (P < 0.01), and also between oral leukoplakia with dysplasia and those without dysplasia (P < 0.01). Wnt3 expression was detected in the epithelial cell membrane or cytoplasm in oral leukoplakia where nuclear expression of beta catenin was evident, but not in epithelial cells without nuclear expression of beta-catenin. CONCLUSION: The components of canonical Wnt pathway, such as Wnt3, beta-catenin, and cyclin D1, were detected, implying that this pathway is potentially involved in the progression of dysplasia in oral leukoplakia. PMID- 17922925 TI - Computational analysis of the relationship between allergenicity and digestibility of allergenic proteins in simulated gastric fluid. AB - BACKGROUND: Safety assessment of genetically modified (GM) food, with regard to allergenic potential of transgene-encoded xenoproteins, typically involves several different methods, evaluation by digestibility being one thereof. However, there are still debates about whether the allergenicity of food allergens is related to their resistance to digestion by the gastric fluid. The disagreements may in part stem from classification of allergens only by their sources, which we believe is inadequate, and the difficulties in achieving identical experimental conditions for studying digestion by simulated gastric fluid (SGF) so that results can be compared. Here, we reclassify allergenic food allergens into alimentary canal-sensitized (ACS) and non-alimentary canal sensitized (NACS) allergens and use a computational model that simulates gastric fluid digestion to analyze the digestibilities of these two types. RESULTS: The model presented in this paper is as effective as SGF digestion experiments, but more stable and reproducible. On the basis of this model, food allergens are satisfactorily classified as ACS and NACS types by their pathways for sensitization; the former are relatively resistant to gastric fluid digestion while the later are relatively labile. CONCLUSION: The results suggest that it is better to classify allergens into ACS and NACS types when understanding the relationship between their digestibility and allergenicity and the digestibility of a target foreign protein is a parameter for evaluating its allergenicity during safety assessments of GM food. PMID- 17922926 TI - Adolescents' experiences of being food-hypersensitive: a qualitative study. AB - BACKGROUND: Experiencing or being at risk of adverse reactions to certain food items is a common health issue, especially among children and adolescents. Research has shown that living with the risk of food reactions and always having to take measures to avoid certain food in one's diet has a negative impact on quality of life. The aim of this study was to illuminate adolescents' experiences of being food hypersensitive. METHODS: Three focus group interviews and six individual interviews were carried out with all together 17 adolescents, 14-18 years of age, who had exclusion diets at school due to food hypersensitivity. The interviews were taped and transcribed verbatim and a qualitative content analysis was carried out. RESULTS: Five categories with subcategories, and one pervading theme, emerged. The categories were: Perceiving oneself as being particular, Feeling constrained, Experiencing others' ignorance, Keeping control, and Feeling it's okay. A pervading theme was conceptualised as Striving to normalise the experience of being food-hypersensitive. The adolescents regarded themselves as competent and courageous, but also described how they avoided the extra attention it implied to ask for special food considerations taken into account. Their self conceptions were probably essential for their management of and attitude toward the hypersensitivity condition. They felt deprived, and those at risk of severe food reactions experienced insecurity and fear. Feelings of being disregarded were expressed, as well as facing unreliability and a lack of understanding from others. The continual work of constant vigilance and decision-making was described as time-consuming and frustrating. However, the adolescents also experienced considerate and supportive surroundings and were at pains to tone down the negative experiences and consequences of being food-hypersensitive. CONCLUSION: Food avoidance by itself, and not only the somatic food reactions, brings about consequences with significant impacts on adolescents' lives. The findings from this study have implications for all of those who deal with adolescents who are food-hypersensitive, and not only health professionals. A deeper insight into adolescents' experiences gives an understanding which can improve the care-givers' efforts. PMID- 17922927 TI - Continuous vs. intermittent hemodialysis: with which spin will my patient win? PMID- 17922928 TI - Anti-Trypanosoma cruzi effects of cyclosporin A derivatives: possible role of a P glycoprotein and parasite cyclophilins. AB - Cyclophilins are target molecules for cyclosporin A (CsA), an immunosuppressive antimicrobial drug. We have previously reported the in vitro anti-Trypanosoma cruzi activity of H-7-94 and F-7-62 non-immunosuppressive CsA analogues. In this work, we continue the study of the parasiticidal effect of H-7-94 and F-7-62 CsA analogues in vitro and in vivo and we analyse 3 new CsA derivatives: MeIle-4-CsA (NIM 811), MeVal-4-CsA (MeVal-4) and D-MeAla-3-EtVal-4-CsA, (EtVal-4). The most efficient anti-T. cruzi effect was observed with H-7-94, F-7-62 and MeVal-4 CsA analogues evidenced as inhibition of epimastigote proliferation, trypomastigote penetration, intracellular amastigote development and in vivo T. cruzi infection. This trypanocidal activity could be due to inhibition of the peptidyl prolyl cis trans isomerase activity on the T. cruzi recombinant cyclophilins tested. Furthermore, CsA and F-7-62 derivative inhibited the efflux of rhodamine 123 from T. cruzi epimastigotes, suggesting an interference with a P-glycoprotein activity. Moreover, H-7-94 and F-7-62 CsA analogues were not toxic as shown by cell viability and by aminopyrine-N-demethylase activity on mammalian cells. Our results show that H-7-94, F-7-62 and MeVal-4 CsA analogues expressed the highest inhibiting effects on T. cruzi, being promissory parasiticidal drugs worthy of further studies. PMID- 17922929 TI - Quantitative analysis of parasite DNA in the blood of immunized and naive mice after infection with Neospora caninum. AB - Real-time PCR was used to study the duration and level of parasitaemia in mice immunized with immune-stimulating complexes (iscoms) containing recombinant NcSRS2, one of the immunodominant surface antigens of Neospora caninum. After challenge infection, blood was collected daily for 9 days. During this period the amounts of parasite DNA detected in immunized mice were significantly lower (P<0.001), and the duration of parasitaemia appeared to be shorter, than in non immunized controls. Furthermore, the degree of parasitaemia seemed to correlate well with the amount of N. caninum DNA in the brain 3 weeks post-inoculation and with disease severity measured as changes in body weight. These results indicate that the protective immunity induced by the NcSRS2-iscoms was sufficient to reduce the level of parasitaemia, which probably reduced the number of parasites reaching the brain, and could be the reason for the reduction in brain parasite load and clinical symptoms. Furthermore, real-time PCR was found to be a sensitive means for rapid assessment of N. caninum in blood. PMID- 17922930 TI - Evidence that primary infection of Charollais sheep with Toxoplasma gondii may not prevent foetal infection and abortion in subsequent lambings. AB - A study carried out on a sheep farm examined whether Toxoplasma gondii foetal infection and associated abortion occur in successive lambings. We identified 29 ewes that gave birth to lambs on at least 2 successive years over our study period, 2000-2003. Tissue samples from the progeny of these ewes were analysed by PCR to determine infection status with T. gondii. T. gondii-infected lambs were born in 31% of successive pregnancies. T. gondii-positive lambs were aborted in successive pregnancies in 21% of lambings during study period, 2000-2003. The frequency of successive abortions within this flock over the period 1992-2003 was 18%. If a lamb was congenitally infected there was a high risk (69%) that the successive lamb from that ewe would also be congenitally infected. Similarly, if a lamb was aborted there was a high risk (55%) of abortion in the next lamb produced. These data suggest that life-long immunity to T. gondii infections may not always be acquired following an initial infection and raises the question as to whether the mechanisms of T. gondii transmission prior to and during ovine pregnancies are fully understood. PMID- 17922931 TI - HIV subtypes in Scotland, 2000-2006. AB - The purpose of this study was to document the dynamics of HIV-1 subtypes in Scotland over a 6-year period. Viral RNA from all-new diagnoses was amplified by nested PCR and sequenced in the gag and/or env regions. Subtype was assigned by phylogenetic analysis, and aligned with demographic data including likely route and geographical origin of infection. We present data on 80% of all new diagnoses in Scotland between April 2000 and April 2006. Within the background of an expanding epidemic, subtype B predominates in men who have sex with men and intravenous drug users but there is a small but consistent number of UK-acquired infections in these risk groups caused by non-B subtypes. In heterosexuals, non-B subtypes acquired abroad, especially Africa, are still the largest group but again UK-acquired numbers are rising. The social and clinical significance of the spread of non-B subtypes in different ethnic and risk groups remains to be established. PMID- 17922932 TI - Identification of newly recognized serotype 1c as the most prevalent Shigella flexneri serotype in northern rural Vietnam. AB - We investigated the identity of 37 Shigella flexneri strains that had previously been isolated from northern rural Vietnam (Son Tay Province) and described as untypable. Twenty-four isolates reacted with MASF 1c, a monoclonal antibody specific for S. flexneri serotype 1c. A further ten untypable isolates were found to be rough mutants (no longer expressing O-antigen) that were derived from serotype 1c strains. Pulsed-field gel electrophoresis demonstrated that these strains consisted of many different clones, indicating serotype 1c was well established in this region in the late 1990s. Serotype 1c was the most prevalent S. flexneri serotype isolated in the Son Tay Province, accounting for about 40% of S. flexneri isolates. Subsequent isolation of S. flexneri serotype 1c in this region and elsewhere in Vietnam confirmed that serotype 1c is of genuine importance in Vietnam. PMID- 17922933 TI - Investigation of the vitamins A and E and beta-carotene content in milk from UK organic and conventional dairy farms. AB - During a 12-month longitudinal study, bulk-tank milk was collected from organic (n=17) and conventional (n=19) dairy farms in the UK. Milk samples were analysed for vitamin A (retinol), vitamin E (alpha-tocopherol) and beta-carotene content. The farming system type, herd production level and nutritional factors affecting the milk fat vitamin content were investigated by use of mixed model analyses. Conventionally produced milk fat had a higher mean content of vitamin A than organically produced milk fat, although there were no significant differences in the vitamin E or beta-carotene contents between the two types of milk fat. Apart from farming system, other key factors that affected milk fat vitamin content were season, herd yield and concentrate feeding level. Milk vitamin content increased in the summer months and in association with increased concentrate feeding, whilst higher-yielding herds had a lower milk vitamin E and beta carotene content. Thus, conventional dairy farms in the UK produced milk with a higher vitamin A content, possibly owing to increased vitamin A supplementation in concentrate feeds. However, knowledge of the effects of season, access to fresh grazing or specific silage types and herd production level may also be used by all producers and processors to enhance the vitamin content in milk. PMID- 17922934 TI - Stochastic modelling to assess economic effects of treatment of chronic subclinical mastitis caused by Streptococcus uberis. AB - Chronic subclinical mastitis is usually not treated during the lactation. However, some veterinarians regard treatment of some types of subclinical mastitis to be effective. The goal of this research was to develop a stochastic Monte Carlo simulation model to support decisions around treatment of chronic subclinical mastitis caused by Streptococcus uberis. Factors in the model included the probability of cure after treatment, probability of the cow becoming clinically diseased, transmission of infection to other cows, and physiological effects of the infection. Using basic input parameters for Dutch circumstances, the average economic costs per cow of an untreated chronic subclinical mastitis case caused by Str. uberis in a single quarter from day of diagnosis onwards was euro109. With treatment, the average costs were higher (euro120). Thus, for the average cow, treatment was not efficient economically. However, the risk of high costs was much higher when cows with chronic subclinical mastitis were not treated. A sensitivity analysis showed that profitability of treatment of chronic subclinical Str. uberis mastitis depended on farm-specific factors (such as economic value of discarded milk) and cow-specific factors (such as day of diagnosis, duration of infection, amount of transmission to other cows and cure rate). Therefore, herd level protocols are not sufficient and decision support should be cow specific. Given the importance of cow-specific factors, information from the current model could be applied to automatic decision support systems. PMID- 17922935 TI - A novel minimal model to describe non-esterified fatty acid kinetics in Holstein dairy cows. AB - The dynamics of non-esterified fatty acid (NEFA) metabolism in lactating dairy cows requires quantification if we are to understand how dietary treatments and disease influence changes in body condition (adipose reserves) and the production of milk fat. We present here a novel compartmental model that employs the pattern of plasma glucose concentrations to predict the dynamic changes that occur in plasma NEFA concentrations during an intravenous glucose tolerance test (IVGTT) in lactating dairy cows. The model was developed using data obtained from ten early-lactation, Holstein-Friesian cows given a standard IVGTT. The model described all of the major features of the NEFA response to an IVGTT; it was consistent with physiological processes and provided a number of parameters that can be used to quantify NEFA production and utilization. For all of the individual cows, all model parameters were well identified and usually had CV<10% of their estimated values. In the model, elevated plasma glucose concentrations cause an increase in the level of glucose in a remote compartment, which in turn suppresses the rate of entry of NEFA to the plasma compartment. The means (+/-sd) for the five adjustable parameters of the model were: rate of entry of NEFA to the plasma pool (SFFA) 183+/-71 [micromol l-1 min-1], rate of removal (oxidation, sequestration in adipose tissue and uptake by the mammary gland for milk production) of NEFA from the plasma pool (KFFA) 0.140+/-0.047 [min-1], a threshold parameter (gs) representing a plasma glucose concentration above which elevated levels of plasma glucose result in entry of glucose into a 'remote' or inaccessible glucose compartment, 3.30+/-0.52 [mmol/l], a rate constant (K) describing the movement of plasma glucose (above gs) into a remote compartment 0.063+/-0.033 [min-1] and a parameter Phi which is a Michaelis Menten type affinity constant which modulates the extent to which remote glucose inhibits the provision of NEFA to the plasma pool, 0.812+/-0.276 [mmol/l]. It is concluded that the model is suitable to describe NEFA kinetics in lactating dairy cows and it may have application in other species. PMID- 17922936 TI - Supplementary feeding at milking and minimum milking interval effects on cow traffic and milking performance in a pasture-based automatic milking system. AB - In extensive pastoral dairy farming systems herds graze 12 months of the year with the majority fed a near-100% pasture or conserved pasture diet. The viability of automatic milking in these systems will depend partly upon the amount of supplementary feed necessary to encourage cows to walk from the pasture to the milking unit but also on the efficient use of the automatic milking system (AMS). This paper describes a study to determine the importance of offering concentrate in the milking unit and the effect of minimum milking interval on cow movement and milking performance in a pasture-based AMS. The effects of feeding rate (FR0=0 kg or FR1=1 kg crushed barley/d) and minimum milking interval (MM6=6 h or MM12=12 h) on cow movement and behaviour during milking were studied in a multi-factorial cross-over (feeding level only, 4 weeks per treatment) experiment involving 27 mixed-breed cows milked through a single AMS. Feeding 1 kg barley in the milking unit resulted in a higher visiting frequency to the pre-selection unit (FR0=4.6 visits/d, FR1=5.4 visits/d, sed=0.35, P<0.05) and a higher yield (FR0=22.5 kg/d, FR1=23.6 kg/d, sed=0.385, P<0.01) but had no effect on milking frequency (FR0=1.6 milkings/d, FR1=1.7 milkings/d, sed=0.04, NS). Minimum milking interval was the major factor influencing milking frequency (MM6=1.9, MM12=1.4 milkings/d, sed=0.15, P<0.01). The absence of feeding in the milking unit had no negative effect on behaviour during milking or the number of cows that had to be manually driven from the paddock. The results show that automatic milking can be combined with a near-100% pasture diet and that milking interval is an important determinant for maximizing milk harvested per AMS. PMID- 17922937 TI - Effect of relocation on locomotion and cleanliness in dairy cows. AB - This study was conducted to determine the effect that relocation to a new free stall barn had on locomotion and cleanliness of two breeds of dairy cows. The original facility before relocation had cows housed in an 8-row free stall barn. Cows were allocated in a new 4-row free stall facility: cows of two breeds (n=22 Holsteins and 22 Jerseys) were intermixed in the northwest section. Locomotion (scale 1-5) and cleanliness were scored (scale 1-4). Holsteins and Jerseys had no difference in locomotion score throughout 12 weeks following relocation. A lactation number by date interaction showed cows in third and greater lactations had significantly higher locomotion scores (more lameness) by day 86. Locomotion scores increased across breeds during the 86-d observation period, suggesting cows became lamer. Jerseys had cleaner lower legs than Holsteins (2.9+/-0.1 v. 3.5+/-0.1, respectively). Lactation number affected lower leg cleanliness, with scores decreasing as lactation number increased (3.4 v. 3.3 v. 2.9+/-0.10 for first, second and third and greater lactations, respectively; P<0.01). All cows were cleaner (lower scores) after relocation, suggesting that the new facility improved hygiene. PMID- 17922938 TI - Meta-analyses of cognitive functioning in euthymic bipolar patients and their first-degree relatives. AB - BACKGROUND: Previous work suggests that impairments in executive function and verbal memory in particular may persist in euthymic bipolar patients and serve as an indicator of genetic risk (endophenotype). METHOD: A systematic review of the literature was undertaken. Effects sizes were extracted from selected papers and pooled using meta-analytical techniques. RESULTS: In bipolar patients, large effect sizes (d>0.8) were noted for executive functions (working memory, executive control, fluency) and verbal memory. Medium effect sizes (0.5 or = 1 liter, baseline performance status of 0/1, and weight loss < 5% in 3 months preceding the trial. Univariate analysis, Cox proportional hazards regression, and parametric accelerated failure time models were performed to identify the factors that affected survival duration. Variables analyzed included age (< 70 years vs. > or = 70 years), sex, ethnicity, body mass index, performance status (0 vs. 1), FEV(1) (> 2 L vs. 1-2 L), smoking status (current vs. never/former), hemoglobin (Hb) level, use of positron emission tomography scan in staging, and stage (IIIA vs. IIIB). RESULTS: Median follow-up was 25.6 months and the median OS was 21.2 months. The univariate analysis showed that Hb levels > or = 12 were associated with an improved survival (P = .033). The multivariable parametric accelerated failure time model demonstrated the association of FEV(1) > 2 L (P = .014), and higher pretreatment Hb values (P = .007) as independent prognostic factors for OS. Similarly in the Cox regression, survival was influenced by Hb and FEV(1) > 2 L. CONCLUSION: This analysis suggests that FEV(1) > 2 L and higher pretreatment Hb values are associated with improved OS in patients with stage III NSCLC. These factors can be useful in predicting for more favorable outcomes in patients with stage III NSCLC and provide additional information when designing future studies. PMID- 17922972 TI - Paclitaxel/Carboplatin/gemcitabine versus gemcitabine/vinorelbine in advanced non small-cell lung cancer: a phase II/III study of the Minnie Pearl Cancer Research Network. AB - PURPOSE: This prospective randomized study compared overall survival (OS) in patients with previously untreated advanced non-small-cell lung cancer (NSCLC) when treated with the platinum agent-based triple drug combination of paclitaxel/carboplatin/gemcitabine (PCG) versus the nonplatinum agent-based doublet drug combination of gemcitabine/vinorelbine. PATIENTS AND METHODS: Advanced (stages IIIB, IV, and recurrent) chemotherapy-naive patients with NSCLC and performance status 0-2 were randomly assigned to the PCG arm (paclitaxel 200 mg/m(2) on day 1, carboplatin area under the concentration-time curve of 5 on day 1, and gemcitabine 1000 mg/m(2) on days 1 and 8, every 21 days) or to the gemcitabine/vinorelbine arm (gemcitabine 1000 mg/m(2) on days 1, 8, and 15 and vinorelbine 25 mg/m(2) on days 1, 8, and 15, every 28 days). RESULTS: A total of 337 patients were randomly assigned to the 2 arms. The median time to progression was 6 months for PCG and 3.9 months for gemcitabine/vinorelbine with 1- and 2 year progression-free survival rates of 13% and 2% versus 14% and 4% (P = .324 log rank). Median OS for PCG was 10.3 months versus 10.7 months for gemcitabine/vinorelbine with 1-, 2-, and 3-year OS rates of 38%, 12%, and 2% versus 45%, 12%, and 6%, respectively (P = 0.269 log rank). Grade 3/4 thrombocytopenia, nausea/vomiting, myalgia/arthralgia, and neuropathy were significantly greater in the PCG arm. CONCLUSION: There was no difference in OS or progression-free survival when comparing PCG and gemcitabine/vinorelbine, and gemcitabine/vinorelbine was significantly less toxic. Gemcitabine/vinorelbine is a reasonable nonplatinum agent-based doublet therapy for patients with advanced NSCLC. PMID- 17922973 TI - CyberKnife radiosurgery for stage I lung cancer: results at 36 months. AB - PURPOSE: The aims of this study were to determine if image-guided robotic stereotactic radiosurgery by CyberKnife Radiosurgery System using ablative radiation doses achieves acceptable local control in medically inoperable patients with early non-small-cell lung cancer (NSCLC) and to evaluate disease free survival, toxicity, and failure. CyberKnife can deliver the prescribed dose by using many different angles converging on the target, with real-time target tracking through a combined orthogonal radiograph imaging and optic motion tracking system (Synchrony). MATERIALS AND METHODS: A review of treatment details and outcomes for 59 patients, ranging in age from 51 years to 96 years, with 61 tumors with histologically proven cancers treated by image-guided robotic stereotactic radiosurgery at the CyberKnife Center of Miami between March 2004 and March 2007 is presented. Target localization and respiratory movement compensation were accomplished using a single fiducial marker placed within the tumor, and the X-Sight and Synchrony systems. Total doses ranged from 15 Gy to 67.5 Gy delivered in 1-5 fractions with an equivalent dose range of 24-110 Gy normalized treatment dose in 2 Gy fractions (alpha/beta = 20 Gy). RESULTS: Four patients with stage 1A NSCLC and 2 patients with stage 1B NSCLC had persistent or recurrent disease. All patients tolerated the radiosurgery well, fatigue being the main side effect. Of the 59 patients treated, 51 (86%) were still alive at 1 33-month follow-up. Eight patients have died, 2 of diseases other than cancer progression. CONCLUSION: The results indicate that the delivery of precisely targeted ablative radiation doses with surgical precision to limited treatment volumes of lung tumors in a hypofractionated fashion is feasible and safe. Image guided robotic stereotactic radiosurgery of lung tumors with CyberKnife(R) achieves excellent rates of local disease control with limited toxicity to surrounding tissues and, in many cases, might be curative for patients for whom surgery is not an option. PMID- 17922974 TI - CCND1 messenger RNA expression is correlated with EGFR mutation status in lung cancer. AB - BACKGROUND: Epidermal growth factor receptor (EGFR) stimulation markedly increases cyclin D1 protein expression. Recently, it has been reported that cyclin D1 expression was increased in EGFR mutant cell lines; however, the expression status of CCND1 in EGFR mutant lung cancer tissues has not been reported. PATIENTS AND METHODS: We have investigated the CCND1 messenger RNA (mRNA) levels and other clinicopathologic data in 74 lung cancers. The CCND1 mRNA levels were quantified by real-time reverse-transcriptase polymerase chain reaction using LightCycler. RESULTS: The CCND1/GAPDH mRNA levels were significantly higher in adenocarcinoma (35.125 +/- 37.387) than in non adenocarcinoma (15.2 +/- 24.699; P = .0158), and CCND1/GAPDH mRNA levels were not significantly different among smoking status, sex, or pathologic stage. The CCND1/GAPDH mRNA levels were significantly higher in lung cancer with EGFR mutation (39.713 +/- 41.265) than in lung cancer without EGFR mutation (21.805 +/ 29.152; P = .0338). CCND/GAPDH mRNA expression did not correlate with prognosis of lung cancer. CONCLUSION: Using the LightCycler real-time reverse-transcriptase polymerase chain reaction assay, CCND1 gene expression might correlate with EGFR mutation in lung cancer. However, further studies are needed to confirm the impact of cyclin D1 for a molecular target of lung cancer. PMID- 17922975 TI - Safety of irinotecan/cisplatin versus etoposide/cisplatin for patients with extensive-stage small-cell lung cancer: a metaanalysis. AB - PURPOSE: The objective of this study was to evaluate the safety of patients with extensive small-cell lung cancer treated with irinotecan/cisplatin (IP) versus etoposide/cisplatin (EP). PATIENTS AND METHODS: This is a metaanalysis of a randomized controlled trial. The main outcome measures for safety were grade 3/4 leukopenia, grade 3 anemia, grade 3/4 thrombocytopenia, grade 3/4 neutropenia, grade 3 vomiting/nausea, grade 3/4 diarrhea, and infection. RESULTS: Three randomized controlled trials totaling 535 patients were included. Metaanalysis results are as follows: fewer patients receiving IP experienced grade 3/4 leukopenia (response rate [RR], 0.44; 95% confidence interval [CI], 0.29-0.67), grade 3 anemia (RR, 0.65; 95% CI, 0.43-0.99), and grade 3/4 thrombocytopenia (RR, 0.23; 95% CI, 0.12-0.42), compared with patients receiving EP. But more patients experienced grade 3 vomiting or nausea (RR, 2.27; 95% CI, 1.37-3.37) and grade 3/4 diarrhea (RR, 21.66; 95% CI, 4.87-96.2). There was no significant difference between the 2 groups with regard to infection (RR, 0.75; 95% CI, 0.54-1.04). CONCLUSION: Current clinical studies might confirm that fewer patients receiving IP experienced grade 3/4 leukopenia, grade 3 anemia, grade 3/4 thrombocytopenia, and grade 3/4 neutropenia, compared with patients receiving EP, but more experienced grade 3 vomiting/nausea and grade 3/4 diarrhea. There was no significant difference between the group receiving IP and the group receiving EP with regard to infection. Although there is convincing evidence to confirm the results mentioned herein, they still need to be confirmed by large-sample, multicenter, randomized, controlled trials. PMID- 17922976 TI - Relief from sleep apnea after radiation and chemotherapy. AB - Superior vena cava syndrome (SVCS) can result from extrinsic compression by a primary tumor, mediastinal lymph nodes metastases, benign lesions, or intraluminal thrombosis. The association between obstructive sleep apnea and SVCS has not been extensively evaluated. To our knowledge, only 5 cases of obstructive sleep apnea in SVCS have been reported in the literature. We presented a 53-year old man who was admitted with dyspnea, edema of the face, and excessive daytime sleepiness. Chest radiography and computed tomography revealed lung cancer. A biopsy of the tumor revealed squamous cell carcinoma. Obstructive sleep apnea was diagnosed by polysomnography (apnea hypopnea index: 13 per hour). After radiation and chemotherapy, edema of the face, snoring, and daytime sleepiness were alleviated, and the patient's apnea hypopnea index decreased to 0.6 per hour. In conclusion, there is a relationship between obstructive sleep apnea and SVCS. PMID- 17922977 TI - Malignant primary pulmonary tumor with hemangiopericytoma-like features: conventional hemangiopericytoma versus solitary fibrous tumor. AB - Although extremely rare, the primitive lung hemangiopericytoma in adults is similar to hemangiopericytomas with other soft tissue localizations. Although generally benign and curable after radical surgery, it might also have a malignant clinical course with dissemination in both lungs, infiltration of vital organs (heart, pulmonary artery), extension to the adjacent tissues, and even pulmonary metastases. The treatment of choice is the complete tumor resection with negative surgical margins after excision. Certain histologic features might indicate a malignant potential. The clinical outcome of patients is variable: some are cured after radical surgery and others might present relapse and recurrences that necessitate a second intervention, radiation therapy, and/or chemotherapy. Over the years, the conventionally-defined hemangiopericytoma concept has evolved because of the nonspecific histologic growth pattern (characteristic monotonous appearance, moderate or high cellularity, and a well developed branching vascular pattern) shared by numerous, unrelated benign or malignant lesions. Hemangiopericytoma is no longer considered a specific entity but rather as a characteristic growth pattern; in the heterogeneous group of hemangiopericytoma-like neoplasms, many lesion categories have been individualized and defined. We report an uncommon case of primitive lung tumor exhibiting hemangiopericytoma-like features, with an aggressive, fatal clinical course. Because of the major histologic overlap between solitary fibrous tumor and hemangiopericytoma and lack of clear classification criteria, we encountered difficulty in including this case in a known clinical entity; primitive solitary fibrous tumor of the lung, which mimics lung hemangiopericytoma, seemed to be the most plausible diagnosis. We discuss the case particularities and the radiologic and pathologic correlations. PMID- 17922978 TI - Mometasone furoate dry powder inhaler: a once-daily inhaled corticosteroid for the treatment of persistent asthma. AB - BACKGROUND: Mometasone furoate (MF), a potent synthetic inhaled corticosteroid (ICS) with a high affinity for the glucocorticoid receptor, is approved for use in the treatment of asthma. SCOPE: Publications reviewed in this article were identified via searches of MEDLINE and EMBASE databases using the terms 'mometasone furoate AND pharmacology' and 'mometasone furoate AND asthma AND clinical trial'. Data from abstracts presented at respiratory society meetings, and relevant background information, are also reviewed. FINDINGS: In clinical studies, MF, administered by dry powder inhaler (MF-DPI), was effective in treating all severities of persistent asthma, improving pulmonary function, reducing asthma symptoms, and reducing or eliminating the need for oral corticosteroids. Once-daily dosing of MF-DPI was effective in patients with mild or moderate persistent asthma previously taking twice-daily regimens of inhaled corticosteroids (ICSs), and in patients taking only inhaled beta2-agonists for symptom relief. Once-daily dosing in the evening with MF-DPI 200 microg conferred a greater benefit than morning dosing with MF-DPI 200 microg. Patients with severe asthma who were dependent on oral corticosteroids (OCSs) and high doses of ICSs were able to achieve greater asthma control and reduce or even eliminate OCSs when switched to MF-DPI. In trials of up to 1 year in duration, MF-DPI was well tolerated, with the majority of adverse events considered mild or moderate in intensity. MF had low systemic bioavailability and no clinically significant hypothalamic-pituitary-adrenal-axis suppression at therapeutic doses. The DPI device is a multiple-dose inhaler with a counter containing agglomerates of MF and lactose. Patients of all severities of persistent asthma were able to generate and maintain airflow profiles necessary to provide a uniform and accurate dose. LIMITATIONS: Only one study evaluated both morning and evening administration of once-daily doses, and one of the comparative clinical trials was an open-label study. CONCLUSION: Once-daily administration of MF-DPI 200-400 microg in patients with mild to moderate persistent asthma effectively improved lung function and asthma control. In patients with severe persistent asthma dependent on oral corticosteroids, treatment with MF-DPI 400 microg BID permitted substantial reduction of oral corticosteroid use. All MF-DPI treatments were well tolerated and had minimal systemic effects. PMID- 17922979 TI - The effect of fixed combination of fluticasone and salmeterol on asthma drug utilization, asthma drug cost, and episodes of asthma exacerbations. AB - OBJECTIVE: This study evaluated the use and drug costs of inhaled corticosteroids (ICSs), long-acting beta2-agonists (LABAs), and fluticasone propionate and salmeterol in a fixed-dose combination (FSC) and their relationship to asthma exacerbations before and after the market introduction of FSC in April 2001. METHODS: This is a retrospective analysis of employer-sponsored health insurance claims filed between January 1, 1998, and December 31, 2003 to detect impact of introduction of FSC (approved by the US Food and Drug Administration in August 2000) on utilization and cost of FSC, any ICS (excluding FSC), and any LABA (excluding FSC) along with utilization of medical services related to asthma exacerbations. Asthma medications were identified using National Drug Codes and Redbook, whereas asthma exacerbations were identified using ICD-9-CM primary diagnosis code 493.x. These medical and pharmacy claims were converted to rates per 100 asthma office visits. RESULTS: For all ICSs, the average pharmacy claims per 100 office visits increased from 383 in the year before FSC was introduced to 407 (120 [29.5%] were for FSC and 287 [70.5%] were for single-entity ICSs) in 2003. LABA prescribing increased from 72 in the year before FSC to 147 (120 from FSC, 27 single-entity LABA) in 2003 (p < 0.001). An additional $13,511 per 100 asthma office visits was spent on the FSC product (p < 0.001). After the introduction of FSC, there was no significant difference in asthma admissions (p = 0.17), whereas emergency department (ED) visits increased by 0.92 visits per 100 office visits (p = 0.03). The diagnosis and severity of asthma was inferred from the pharmacy claims and patients with chronic obstructive pulmonary disease could not be excluded. In addition, the study was not designed to assess the impact of other asthma medications on the disease and/or associated costs, and patient adherence to claimed medication could not be monitored. CONCLUSIONS: The introduction of FSC was associated with increased LABAs/FSC patient exposure and expenditure with no change in asthma hospitalizations and an increase in ED visits. PMID- 17922980 TI - Estimated first-year costs of prostaglandin analogs with/without adjunctive therapy for glaucoma management: a United States perspective. AB - OBJECTIVE: To evaluate costs associated with prostaglandin analogs among newly diagnosed glaucoma patients in a managed-care population. RESEARCH DESIGN AND METHODS: A cost minimization model compared annual costs for patients initiating therapy with one of the three prostaglandin analogs (bimatoprost, latanoprost, travoprost). The study cohort was identified from pharmacy claims and eligibility files of patients who met study inclusion criteria. Annual costs were estimated for patients initiating therapy with each prostaglandin based on treatment patterns and medication use over the year. Costs for outpatient physician visits and medications were estimated from standard sources. RESULTS: A total of 4444 patients met study criteria: 674 received travoprost, 729 received bimatoprost, and 3041 received latanoprost. More than 80% stayed on monotherapy for 1 year (82.9% of travoprost patients, 82.8% of bimatoprost patients, and 80.5% of latanoprost patients). Of those who required adjunctive therapy, the average number of days until starting adjunctive therapy was 130 days for travoprost patients, 94 days for bimatoprost patients, and 104 days for latanoprost patients. Average annual costs were $1198, $1290, and $1217 for patients treated with travoprost, bimatoprost, and latanoprost, respectively. CONCLUSIONS: The use of adjunctive therapy in glaucoma is an important driver of glaucoma management costs. This study demonstrates that the longer duration of monotherapy and the likelihood to use single rather than combination adjunctive agents contribute to lower annual costs among patients starting on travoprost compared with the other available prostaglandin analogs. Study limitations include the lack of clinical indicators in the study database; further, results may not be generalizable to patients who discontinue prostaglandin analogs or to the population of patients with glaucoma as a whole. Future studies with clinical and compliance indicators would further identify distinctions among treatment regimens. PMID- 17922982 TI - [Prevention of diabetic foot should be strengthened]. PMID- 17922988 TI - [Study on the biological function of dermal fibroblasts in the wounds of diabetic and non-diabetic rats with deep burns]. AB - OBJECTIVE: To explore the changes of the biological function of dermal fibroblasts (FBs) in the wounds of diabetic and non-diabetic burned rats and the pathogenesis of impaired wound healing in diabetes. METHODS: 80 Sprague-Dawley (SD) rats weighing 220 g were randomly divided into control and STZ-induced diabetic groups, and then deep partial thickness scald involving 10% TBSA was reproduced in the two groups. The diabetic groups were randomized into pre scalding, post-scalding day (PSD 3), PSD 7, PSD 14 and PSD 21 groups, with 6 rats in each group. Controls were also randomized into 5 groups. Skin specimens from the wound were harvested at each time point. Cell cycles of FBs were analyzed with flow cytometry. The amount of hydroxyproline in the skin tissue was assessed on 0, 3, 7, 14, and PSD 21. The type I and III collagens were determined by ELISA. The expression of alpha-SMA in the dermal fibroblasts of each group was assessed by immunohistochemistry method. Transmission electron microscopy was used to observe the ultrastructure changes of FBs. RESULTS: Compared with that in the normal rats, the percentage of the cells in G(0)/G(1) phase in the DM group was evidently lower on PSD 0 (65.79 +/- 5.24 vs 82.43 +/- 9.68, P < 0.01). After the scalding, the percentage of the cells in G(0)/G(1) phase in DM group was significantly higher (70.00 +/- 4.27 vs 42.04 +/- 12.96, on PSD 3, P < 0.01), meanwhile the percentage of S phase was remarkably lower than those in C group on 3, 7, 14, 21PSD (P < 0.05, P < 0.01). The amount of hydroxyproline in the diabetic skin tissue was obviously lower than those of the responding control groups before (0.72 +/- 0.06 vs 1.42 +/- 0.28, P < 0.01) and after burn injury (P < 0.01). Furthermore, the rate of I/III collagen on 7, 14 and PSD 21 was much higher in DM group than that in C group (P < 0.01). The expression of alpha-SMA in DM groups on PSDS 3, 7, 14 and PSD 21 was evidently lower than those of the controls (levels 10.28 +/- 3.99, C group 28.42 +/- 2.73, on PSD 14, P < 0.01), although that inclined to be heightened after burn injury. Ultrastructure changes of FBs in the wounds of diabetic rats could be observed, such as the outstretched endoplasmic reticulum, un development of Golgi's body, lackness of microtubule and microfilament, a sharp increase of cytolysosomes, and so on. CONCLUSION: The FB proliferation in the diabetic skin is abnormal, the synthetical ability of collagen is weakened, the expression of alpha-SMA is insufficient, the microtubule and microfilament is lack, and the number of cytolysosomes increases. The pathogenesis of impaired-wound healing in diabetics might be related with the above mentioned factors. PMID- 17922981 TI - Efficacy and safety of valsartan 160/HCTZ 25 mg in fixed combination in hypertensive patients not controlled by candesartan 32 mg plus HCTZ 25 mg in free combination. AB - OBJECTIVE: Free combination hypertension medication is associated with a lower compliance and less persistence compared to fixed combination therapy and can, therefore, be associated with insufficient blood pressure reductions. This non randomized study investigated whether valsartan 160 mg/hydrochlorothiazide 25 mg (Val 160/HCTZ 25) in fixed dose combination could provide additional blood pressure control in hypertensive patients not adequately controlled by the free combination of candesartan 32 mg plus HCTZ 25 mg. RESEARCH DESIGN AND METHODS: One hundred and ninety-seven patients with a mean sitting diastolic blood pressure (MSDBP) between 100 and 110 mmHg entered a 4-week treatment phase with 32 mg of candesartan in free combination with 25 mg of HCTZ once daily. One hundred and thirty-eight patients with uncontrolled BP at Week 4, entered a second 4-week treatment phase with Val160/HCTZ 25 once daily. MAIN OUTCOME MEASURES: The primary efficacy parameter was the reduction in MSDBP at trough between Week 4 and Week 8 in the intent-to-treat population. RESULTS: At baseline, MSDBP was 103.0 +/- 2.8 mmHg. After Week 4, MSDBP had decreased to 93.8 +/- 4.5 mmHg. Subsequent treatment with Val 160/HCTZ 25 for 4 weeks reduced MSDBP to 88.7 +/- 8.6 mmHg. This represented an additional decrease in MSDBP of 5.1 +/- 7.9 mmHg (p < 0.0001). Val 160/HCTZ 25 reduced mean sitting systolic BP by 3.4 +/ 13.0 mmHg (p = 0.0029). CONCLUSIONS: The fixed dose combination of valsartan 160/HCTZ 25 mg provided a statistically and clinically significant additional BP reduction in patients not controlled by the free combination of candesartan 32 mg and HCTZ 25 mg. PMID- 17922990 TI - [Percutaneous transluminal angioplasty in treatment of lower limb peripheral arterial disease in diabetes patients: analysis of 150 cases]. AB - OBJECTIVE: To analyze the technical success rate and short-term clinical results of percutaneous transluminal angioplasty (PTA) in treatment of lower limb peripheral arterial disease (PAD) in diabetes mellitus (DM) patients. METHODS: 150 consecutive DM patients PAD (176 limbs) with Fontaine degree I - IV were treated with PTA and followed up for 5 months (1 - 16 months). The result with the residual stenosis < 30% was regarded as success. RESULTS: Totally 187 PTA procedures were performed in 150 patients (176 limbs) with a technical success rate of 91.4% (161/176)). No serious complication occurred. The clinical symptoms of all patients improved after successful PTA. The median ankle-brachial index (ABI) marginally increased from the baseline value of 0.34 +/- 0.27 to 0.96 +/- 0.24 after intervention. The ABI levels 1 month and 6 months after PTA were 0.88 +/- 0.34 and 0.71 +/- 0.26 respectively. The cumulative restenosis rate was 13.7%, and the reintervention rate was 7.2%. CONCLUSION: With high success rate, low complication rate, and sure curative effect, PTA can be performed as the first choice in treatment of the lower limb PAD in DM patients. PMID- 17922989 TI - [Differences in risk factors of diabetic foot in the patients in South and North China]. AB - OBJECTIVE: To investigate the differences on the diabetic foot problems and its risk factors in south and north of China. METHODS: Patients with foot problems were surveyed from January 1 to December 31, 2004 in 14 teaching hospitals located in different cities in China, including demographic data, present and past history of the foot problems and peripheral artery disease (PAD), the classification of the foot ulcers based on the Wagner' system, control of the hyperglycemia and lipids disorder, medical cost in hospital and the diabetic complications. All the patients were divided into two groups due to their geographical data, south and north. RESULTS: There were 285 and 349 patients for the group south and group north. No significant differences were found for duration of diabetes or foot problems, fasting or post-meal glucose, total cholesterol, triglycerides, HDL-C, and the numbers of patients with smoke, hypertension, nephropathy or neuropathy between the two groups. There were significant differences for the age (70 yrs vs 66 yrs), percentage of the patients with average person income with over RMB 1000 per month (57.7% vs 45.6%), coronary heart disease (42.6% vs 61.0%) and retinopathy (35.7% vs 49.5%), HbA1c (7.90% vs 8.80 %), LDL-C (2.75 mmol/L vs 2.98 mmol/L), WBC (6.70 x 10(9) vs 7.40 x 10(9)/L), HCT (0.37 vs 0.38), creatinine (87 micromol/L vs 76 micromol/L) and uric acid (333 mmol/L vs 271 mmol/L), and amputation rate (2.6% vs 9.7%) between south and north groups. Logistic analysis showed that severity of the foot problems was associated with ABI and WBC in south group, and with ABI, PLT and HCT in north group. CONCLUSION: Diabetic foot problems were more severe, with more risk factors and with more medical cost in north patients. PMID- 17922991 TI - [Study of foot plantar pressure in Chinese diabetic patients]. AB - OBJECTIVE: To investigate the difference and distribution of plantar pressure in normal Chinese population and diabetic patients and their influencing factors. METHODS: 1035 diabetic patients and 1022 normal subjects were evaluated by the "first step approach". Each trial included five acceptable separate barefoot walks across the EMED-AT system for each foot. The whole foot was divided into ten "masks", including the heel, midfoot, the first, second, third, fourth, fifth metatarsals, hallux, second toe and the third-to-fifth toes. The mean of maximum peak pressure (MPP) of each mask as calculated. RESULTS: The MPP were 410 +/- 122 and 427 +/- 128 kPa in left and right foot of diabetics while 415 +/- 113 and 428 +/- 116 kPa in left and right foot of normal subjects with P > 0.05. The top three places with the hight MPP value in the in diabetics and normal subjects were hallux > the second metatarsal > the third metatarsals in both left and right foot. The disease course, sex, age, BMI, WHR and history of foot ulcer had few influence on the plantar pressure in diabetics. The MPP in the diabetics with peripheral neuropathy (DPN) was higher than that of normal subjects, with no significant difference compared to that of non-DPN diabetics. CONCLUSION: The plantar pressure in the diabetics is almost the same as that of the normal subjects, but with different distribution is different. Complication with DPN may lead to the increase of plantar pressure in the diabetic. But factors such as disease course, sex, age, BMI and WHR have few influence on MPP. PMID- 17922992 TI - [Research on the clinical features and effective factors of 249 diabetic patients with deep foot infection]. AB - OBJECTIVE: 249 diabetic patients with deep foot infection were retrospectively studied. Their clinical features and effective factors related to wound healing were analyzed. METHODS: 249 patients team (physician, surgeon and diabetes specific nurse) were divided into 3 groups. Group A: patients healed without amputation (n = 107), group B: patients healed after amputation (n = 114), and group C: patients didn't heal after amputation. All patients' clinical features, lab examinations and foot wound features were compared. RESULTS: The group A patient were (59 +/- 12) years old, significantly younger than the group B (67 +/ 11, P < 0.01). Hypersensitive C reactive protein (hs-CRP, 18 +/- 5 mg/L) and plasma albumin (32 +/- 7 g/L) of group A were significantly higher than those of group B (13 +/- 5 mg/L and 29 +/- 5 g/L, respectively, P < 0.01). The duration of diabetes mellitus of group B (17 +/- 11) year was significantly longer than that of group A (10 +/- 6 year, P < 0.05). The possibility of probing bone, purulent secretion, necrosis, bone exposure, cacosmia, edema and critical limb ischemia of group B were more frequent (P < 0.01 or P < 0.05) compared to group A. The age of group B (67 +/- 11) are younger than that of group C (72 +/- 9, P < 0.05). In group B, temperature (38.1 +/- 1.1) degrees C, white blood cell (WBC) count (10 +/- 3) x 10(9)/L and hs-CRP (13 +/- 5) mg/L were higher than those of group C (37.4 +/- 0.8 degrees C, 8 +/- 2 x 10(9)/L and 7 +/- 6 mg/L, respectively, all P < 0.05). Critical limb ischemia of group B (37%) was more frequent than that of group A (7%, P < 0.05), but less frequent than that of group C (77%, P < 0.01). hs-CRP and plasma albumin were protective factors for wound healing. Age, the possibility of probing bone, purulent secretion, necrosis, bone exposure, cacosmia, edema and critical limb ischemia were risk factors for wound healing. CONCLUSION: diabetic patients with deep foot infection are difficult to be diagnosed in early stage since they often have no significant clinical syndrome such as fever, redness, swelling and pain, and their WBC count does not increase. Though multidisciplinary team manages these patients, more than half of them need amputation. Wound healing in the patients is related to multiple factors, including age, duration of diabetes mellitus, hs-CRP, plasma albumin, WBC count, level of limb ischemia and wound features. PMID- 17922993 TI - [The prospective study of the clinical features and outcome of HBeAg-negative and HBeAg-positive cirrhosis in patients with chronic type B hepatitis]. AB - OBJECTIVE: Comparing clinical features and prognosis between HBeAg-negative and HBeAg-positive cirrhosis in patients with chronic type B hepatitis. METHODS: 217 chronic type B hepatitis with cirrhosis patients were prospectively studied for 35 months (3 - 47 months). Comparing clinical features and prognosis between HBeAg-negative and HBeAg-positive cirrhosis in patients with chronic type B hepatitis. RESULTS: The numbers of HBeAg-negative cirrhosis in patients with chronic type B hepatitis were more than HBeAg-positive cirrhosis; The median ALT and AST levels of HBeAg-negative patients were lower than HBeAg-positive patients; The median WBC, HGB and PLT levels of HBeAg-negative patients were lower than HBeAg-positive patients; HBV DNA positive rate and proportion of HBV DNA > 10(5) copies/ml of HBeAg-negative patients was lower than HBeAg-positive patients; The mortality rate of HBeAg-negative patients was higher than HBeAg positive patients; among HBeAg-negative patients group, the presence rate of ascite, portal hypertensive gastrointestinal bleeding and HCC of patients treated with lamivudine were lower than the other patients, the proportion of non presence of complications patients treated with lamivudine were higher than the other patients, the proportion of presence of one-two complications patients treated with lamivudine were lower than the other patients; among HBeAg-positive patients group, the presence rate of ascite of patients treated with lamivudine were lower than the other patients, the proportion of non-presence of complications patients treated with lamivudine were higher than the other patients. CONCLUSION: Among the liver cirrhosis patients, the numbers of HBeAg negative cirrhosis were more than HBeAg-positive cirrhosis; HBeAg-negative patients with cirrhosis have lower liver inflammation activity; HBeAg-negative patients with cirrhosis have lower WBC, HGB and PLT levels; HBV DNA positive rate and proportion of HBV DNA > 10(5) copies/ml of HBeAg-negative patients was lower than HBeAg-positive patients; The mortality rate of HBeAg-negative patients was higher than HBeAg-positive patients; Lamivudine treatment is beneficial in HBeAg negative and HBeAg-positive cirrhosis patients. PMID- 17922994 TI - [The histopathologic and clinical analysis of viral chronic hepatitis patients with negative serological viral markers]. AB - OBJECTIVE: To analyze the histopathological and clinical features of viral chronic hepatitis patients with negative serological viral markers. METHODS: 62 hepatitis patients with negative serological markers were assayed with serological viral hepatitis markers, liver function test and liver biopsies were enrolled in the study. Serum HBV DNA of HBV cases was analyzed by PCR. Liver specimens were examined by immunohistochemistry for HBsAg and HBcAg. RESULTS: The fit rate of histopathological diagnosis with clinical diagnosis is 53.2%, the fit rate is 69.1% in moderate chronic hepatitis group. The immunohistochemistry showed that HBsAg and/or HBeAg positive rate was 45.2%, 53.6% had moderate chronic hepatitis and 25% had mild hepatitis. 13 (46.4%) had G1 hepatitis, 10 (35.7%) had G2 hepatitis, 3 (10.8%) had G3 hepatitis and 2 (7.1%) had G4 hepatitis, and serum HBV DNA positive rate was 35.7%. There were no differences in HBV DNA levels between different hepatitis group and fibrosis stage group (P > 0.05). There were no differences in all indexes between HBV DNA negative group and HBV DNA positive group (P > 0.05). There were no differences in all indexes between HBV patients and other patients (P > 0.05). CONCLUSION: Occult HBV infection may account for a high proportion of the cases with chronic hepatitis of unknown etiology. Most patients are chronic mild hepatitis, but they still have HBV replication and can progress to liver cirrhosis. Serum PCR test, liver biopsy and immunohistochemistry are helpful for the diagnosis. PMID- 17922996 TI - [Study of the relapse of patients with chronic hepatitis B undergoing first and repeated recombinant interferon-alpha therapy during long-term follow up]. AB - OBJECTIVE: To investigate the relapse of patients with chronic hepatitis B (CHB) undergoing first and repeated recombinant interferon-alpha (rIFN-alpha) therapy during long-term follow up. METHOD: Five hundred and twenty three patients with chronic hepatitis B including 403 hepatitis B e-antigen (HBeAg) positive patients and 120 HBeAg negative ones were treated with 5MU recombinant interferon-alpha 1b (rIFN-alpha1b) subcutaneously thrice weekly for 6 - 25 (median 10) months. For each patient, serum alanine aminotransferase (ALT) was measured biochemically and serum HBV DNA level was detected by fluorescent-quantitative PCR, HBeAg with enzyme immunoassay every 1 - 3 month during therapy and every 3 - 6 month during the follow up period. Some of the individuals who relapsed during the follow-up period were treated with interferon-alpha repeatedly. RESULTS: Ratios of early response to interferon-alpha were similar in HBeAg positive patients (55.8%, 225/403), and HBeAg negative patients (64.2%, 77/120) at the end of naive treatment (chi(2) = 2.633, P = 0.105). 39.4% (119/302) of early responders relapsed during 39 +/- 22-month follow up, and relapse rates in HBeAg negative group (55.8%, 43/77) were higher than those in HBeAg positive group (33.8%, 76/225) at the end of follow up (chi(2) = 19.335, P = 0.000). Divided the follow up period into six fragments as 1 - 12 months, 13 - 24 months, 25 - 36 months, 37 - 48 months, 48 - 60 months and > or = 61 months, we found that the differences of relapse incidence were significant (chi(2) = 73.518, df = 5, P = 0.000), and accumulative relapse rates were significant too (chi(2) = 32.167, df = 5, P = 0.000) in all follow-up periods. Constituent ratios of HBeAg in relapsed patients of every follow-up period were similar. 57 relapsed individuals (25 in HBeAg positive group and 32 in HBeAg negative group) were retreated with interferon alpha, and complete response were achieved in all cases at the end of repeated therapy. The relapse rates in HBeAg positive group (52.0%, 13/25) were higher than in HBeAg negative group (21.9%, 7/32) during the follow-up period after the end of retreatment (chi(2) = 5.592, P = 0.018). CONCLUSION: Rates of early response to interferon-alpha therapy were similar in HBeAg positive and HBeAg negative patients at the end of nave treatment, and relapse rates in HBeAg negative group were higher than in HBeAg positive group during long term follow up. Combined response was achieved in all relapse cases received repeated interferon-alpha therapy at the end of retreatment. The relapse rates in HBeAg positive group were higher than in HBeAg negative group during the follow-up period after repeated therapy. PMID- 17922997 TI - [Correlation of expression of epidermal growth factor receptor-2 expression and tumor marker p53 in primary breast cancer]. AB - OBJECTIVE: To observe the correlation between expression of epidermal growth factor receptor (HER)-2 and tumor marker p53 in breast cancer. METHODS: specimens of primary breast cancer were collected from 2996l patients, all female, aged 50 (15 - 92). All of the cases were confirmed by pathohistological method. Immunohistochemistry was used to detect the expression of estrogen receptor (ER), progesterone receptor (PR), Her-2, and p53. RESULTS: 680 cases (22.7%) showed HER 2 overexpression, among which 378 cases (12.6%) were HER-2 weak positive (++) and 302 cases (10.1%) were HER-2 strong positive (+++). 935 cases were p53 positive and 2061 cases were p53 negative. The HER-2 overexpression rate was 32.3% in the p53-positive patients and 18.4% in the p53-negative patients. Expression of HER-2 was positively correlated with expression of p53 (P < 0.01). CONCLUSION: Expression of p53 is the independent index of HER-2 expression, and there is a positive correlation between expression of HER-2 and expression of p53. PMID- 17922998 TI - [Characteristics in Raman spectrum of serum of gastric cancer patients]. AB - OBJECTIVE: To investigate the characters of Raman spectrum in gastric cancer patients through comparing serum of functional dyspepsia patients, serum of gastric cancer patients with SGC7901 cell culture fluid, correlation with culturing time of cell line, and research the recourse of these special Raman peak. METHODS: Peripheral blood samples were collected from 25 gastric cancer patients and 10 functional dyspepsia (FD) patients. Human gastric cancer cells of the line SGC7901 were cultured. The serum samples of the gastric cancer patients and of the FD patients, and the culture fluid of SGC7901 cell underwent Raman spectroscopy with 230 nm and 100 nm as the least peaks respectively. Serum of normal person, lysate, and 1604 culture fluid were used as controls. RESULT: Seven reproducible Raman peaks at the wavelengths of 583, 633, 656, 674, 707, 773 and 799 nm were detected, with 230 nm as the least peak, in the serum samples of gastric cancer patients. Raman peaks at the wavelengths of 633, 656, and 674 nm were detected only in the serum samples of gastric cancer patients, and the Raman peaks at the wavelengths of 533, 707, 773, and 799 nm of the serum of gastric cancer patients (96.5 +/- 6.0, 76.9 +/- 4.7, 63.7 +/- 4.5, 285.7 +/- 18.6) were all remarkably higher than those of the serum samples of FD patients (40.3 +/- 1.8, 27.9 +/- 1.9, 22.9 +/- 1.4, 113.2 +/- 7.7, all P < 0.01). Similar condition, with 100 nm as the least peak, could detected in the Ramon peaks at the above mentioned wavelengths between the serum samples of the gastric cancer patients and the 1640 culture medium. The Raman peak of the cell culture fluid was positively correlated with the time of cell culture. The Raman peaks of the gastric cell lysate at the wavelengths of 583, 633, and 656 nm were significantly different from those of the cell culture fluid (P = 0.027, 0.029, and 0.043 respectively). CONCLUSION: The serum of gastric cancer patients has specific reproducible Raman spectrum. The cell content possibly is part of the reason of the special characters in Raman spectrum of gastric cancer serum. PMID- 17922999 TI - [Dissection of prostate coalesce in treatment of benign prostate hypertrophy with relatively small volume]. AB - OBJECTIVE: To evaluate the curative effect of dissection for coalesce of prostate in treatment of bladder outlet obstruction (BOO) caused by benign prostate hypertrophy (BPH) with relatively small volume (less than 30 g). METHODS: Thirty six patients of BOO caused by BPH with relatively small volume underwent dissection of prostate coalesce. Follow-up was conducted for 32 months (9 - 52 months). The values of international prostate symptom score (IPSS), maximum flow rate (Qmax), and postvoid residual volume (PRV) before and after treatment were measured and compared. RESULTS: The post-operative IPSS was 5.03 +/- 2.66, significantly lower than that before operation (14.19 +/- 5.35), the post operative Qmax was (17.71 +/- 4.1) ml/s, significantly higher than that before operation [(6.19 +/- 2.14) ml/s], and the post-operative PRV was (8.53 +/- 4.78) ml, significantly lower than that before operation [(50.58 +/- 14.84) ml] (all P < 0.001). CONCLUSION: Dissection of prostate coalesce is an ideal method in treatment of BOO caused by BPH with relatively small volume (less than 30 g). PMID- 17923000 TI - [Study of intracerebral focus changes on CADASIL by using MR imaging]. AB - OBJECTIVE: To investigate dynamic changes of intracerebral focus on cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). METHODS: Using the modified Scheltens scale, the magnetic resonance (MR) changes of lesion distribution, size and shape in 7 patients from a CADASIL family were retrospectively analyzed during 3 years observed. RESULTS: In 6 of 7 patients, the number and volume of lesion areas in the white matter were increased (parietal lobe, n = 6; temporal lobe, n = 5; frontal lobe, n = 3; occipital lobe, n = 2) and some areas even became confluent as a mass or chain. New lacunar infarcts (n = 1 - 5) appeared in 5 patients and the arcuate fiber were involved in 1 patients. Slight enlargement could be seen in lateral ventricle (n = 1) or lateral ventricle with third ventricle (n = 1). CONCLUSION: MR imaging can help us to reveal dynamic changes of brain lesions and prognosis in patients with CADASIL. PMID- 17923001 TI - [Perlecan participates in the protection of ischemic myocardium induced by basic fibroblast growth factor: experiment with rats]. AB - OBJECTIVE: To investigate if perlecan (PN) is involved in the myocardial protection of basic fibroblast growth factor (bFGF) in acute myocardial infarction. METHODS: Twenty-four Wistar rats were randomized into 3 groups: myocardial infarction group (MI group, n = 8, undergoing ligation of the descending anterior branch of the left coronary artery), bFGF + MI group (n = 10, injected with bFGF into the border myocardium between the infracted and non- infracted areas immediately after the ligation of the descending anterior branch), and sham operation group (n = 6, undergoing sham operation and injection of normal saline). 24 h, 14 days, and 28 days after the operation the hemodynamic parameters, infarct size, and microvessel density (MVD) were observed. The hearts were taken out, RT-PCR was used to detect the mRNA expression of PN, and Western blotting was used to detect the expression of focal adhesion kinase (FAK) and p38 mitogen-activated protein kinase (MAPK) phosphorylation. RESULTS: In the bFGF + MI group, the +dp/dt and -dp/dt were improved markedly, the infarct size was significantly smaller, the MVD value was higher than those of the MI group on day 14 and day 28; the perlecan mRNA expression was higher in the infarct marginal area and interior zone; and the expression of FAK and the p38MAPK phosphorylation were up-regulated in the marginal zone of the bFGF group. CONCLUSION: bFGF is useful in promoting ischemic myocardial angiogenesis; reducing the size of infracted myocardium, and improving the ventricular function in acute myocardial infarction. Perlecan participates in the cardiac protection induced by bFGF. The relevant pathway is related to up-regulation of FAK and p38MAPK. PMID- 17923002 TI - [Development and application of quality control materials for human papillomavirus types 16 and 18 nucleic acid test]. AB - OBJECTIVE: To develop quality control materials for human papillomavirus (HPV) types 16 and 18 molecular detection and to evaluate the applicability of the materials in external quality assessment (EQA) of HPV-16, 18 clinical detection. METHODS: The target gene from HPV16 was cloned into the pEGFP-C1 plasmid, then the recombinant plasmid pEGFP-C1-HPV16 was transfected into the HPV18 carrying HeLa cells. The cultured epithelial cells carrying both HPV16 target gene and HPV18 were collected and fixed using methanol. EQA samples for HPV types 16 and 18 test diluted to the concentrations 1:10, 1:50, 1:100, 1:500 were prepared from the above prepared cells and distributed to 73 EQA participants nationwide to undergo RT-PCR. The reports from the participants were summarized and evaluated. Three samples were blindly mailed to Urumqi, Hohhot, and Xiamen respectively to observe the influence of post. RESULTS: Forty-four of the 73 PCR laboratories sent feedback. For the quality control materials of the concentrations 1:10, 1:50, 1:100, and 1:500 the corresponding Ct values were 29.10, 31.19, 32.15, and 32.73 respectively for HPV-16, and 30.32, 32.13, 32.22, and 35.55 respectively for HPV-18. Stability test indicated that the quality control materials were stable at least for 40 days when stored at 4 degrees C. The EQA data from 44 participants showed, that the average fit rate was 95.1% for the high concentration positive samples and was 57.4% for the low concentration samples, and was 98.3% for the negative samples. No significant changes were detected by real-time PCR in the returned EQA samples that were blindly mailed to Urumqi, Hohhot, and Xiamen. CONCLUSION: A quality control materials for HPV types 16 and 18 molecular detection has been developed, and quality assessment verifies its applicability in EQA. PMID- 17923006 TI - [Future of the development of medicine: from genomics to integrative medicine]. PMID- 17923007 TI - [Incidence, time trend, survival, and predictive factors of lung cancer in Shanghai populations]. AB - OBJECTIVE: To investigate the incidence, survival, and influential factors of lung cancer in the Shanghai populations. METHODS: Data of individual cases of lung cancer 2000 - 2004 were obtained from the Shanghai Cancer registry System. The annual prevent change (APC) of incidence of lung cancer from 1972 to 2004 was calculate by crude and standardized rates so as to analyze the time trends. Cox proportional risk model was used to analyze the factors influencing the survival. RESULTS: 23,196 new cases of lung cancer were diagnosed 2002 - 2004. The crude incidence rate of lung cancer in the females was 33.73/100,000, and the age adjusted rate was 30.90/100,000. The crude incidence rate of lung cancer and the age-adjusted rate for the males were 81.65/100,000 and 33.73/100,000 respectively, both higher than those of females. The APC values of crude incidence 2004 for both the males and females increased by 1.723% and 2.036% respectively in comparison with the values 1972 (both P < 0.01). The age-adjusted APC of standardized incidence rate for the males in the city center was -0.605% (P < 0.01), showing a tendency to reduce; while the age-adjusted APC of standardized incidence rate for the females in the city center was -0.136 (P > 0.05). The proportion of stage IV cases in the females was 47.5%, significantly higher than that in the males (40.0%, P < 0.05). The proportion of adenocarcinoma in the females was 86.1%, significantly higher than that in the males (47.8%, P < 0.05). The 3-year survival rate and median survival time of adenocarcinoma in the females were 30.38% and 1.48 years respectively, both significantly higher than those in the males (22.66% and 0.98 years, both P < 0.01). Female gender, being younger, living in urban area, squamous cancer, early stage, visit to higher grade hospital were the factors beneficial to the prognosis of lung cancer. CONCLUSION: The age adjusted incidence rate of lung cancer in Shanghai is much closer to that of Western Europe and North America. The beneficial factors for higher survival rate are female, younger age, urban residency, squamous cell lung cancer, earlier stage of diagnosis, and higher grade hospital for treatment. Women have statistically better outcomes than men in different stages of disease. PMID- 17923008 TI - [Early and middle outcomes of total arterial revascularization using exclusively internal mammary artery and radial artery]. AB - OBJECTIVE: To evaluate the early and mid-results of the use of the radial artery alongside the internal mammary artery for complete arterial revascularization in coronary bypass surgery. METHODS: From January 1999 to January 2005, 123 patients with coronary artery disease with lesions in more than one branch (target vessel), 114 males and 9 females, aged 51.9 +/- 9.4 (32 approximately 74) underwent coronary artery surgery after the complete arterial revascularization pattern. Before the operation the left ventricular ejection fraction (EF) value ranged 30% approximately 78%, and 24 patients (21.1%) had poor ejection fraction (with the EF < 50%). Sixty-four patients (52%) had previous myocardial infarction. Four patients with unstable angina received urgent operation. 36 patients (29.3%) had disease of the left main trunk. 74% of the patients underwent off-pump bypass technique. The mean number of distal anastomoses was 2.6 +/- 0.4 per patient. 20 patients were followed up for 62 +/- 12.8 months (36 approximately 80 months). RESULTS: The early mortality was 0.8%. Postoperative complications included arterial fibrillation in seven patients (5.7%). One patient underwent IABP insertion because of low cardiac output, 3 patients were re-admitted because of heart failure, angina and arrhythmias. Three patients died of cardiac events. One patient underwent redo-CABG due to recurrent myocardial infarction and one patient underwent percutaneous coronary intervention because of stenosis of the arterial graft. CONCLUSIONS: Complete arterial revascularization using radial and mammary arteries provides excellent early results, showing a low mid-term complication rate and surgical morbidity. PMID- 17923009 TI - [Total knee arthroplasty by lateral parapatellar approach for valgus knee]. AB - OBJECTIVE: To investigate the effect of lateral parapatellar approach in total knee arthroplasty (TKA) of valgus knee. METHODS: Lateral parapatellar approach of total knee arthroplasty was applied in 8 patients (10 knees) with severe valgus osteoarthritis knee (bilateral in 2 cases and unilateral in 6 cases), with the valgus angle > 15 degrees , 1 male (1 knee) and 7 females (9 knee), aged 68.2 (58 - 79), 7 cases (9 knees) being of the Krackow type I and 1 case (1 knee) of the Krackow type I, I. After incision of the skin through lateral knee, ilio-tibial band was prolonged by apple pie arthroplasty. The joint capsule was cut open laterally 2 - 4 cm from the para-patellar edge. Soft tissue balance was performed by releasing I - T band in Gerdy tubercle, lateral collateral ligament and poster lateral capsule from the femur and tibial side. Valgus angle of distal femur cutting were five degree. Whiteside line and trans-epicondylar line were used as AP rotational cutting reference. All patellar of the group were resurfaced. Capsule closure is completed with the knee flexed. The expanded deep lateral soft tissue sleeve (coronal Z-plasty) is sutured with the medial retinaculum sleeve (superficial layer). Follow-up was conducted for 19.6 months (1 - 51 months). RESULTS: Seven cases (9 knees) were replaced by posterior stabilized cemented prostheses (TC-Dynamic, PLUS), one case (1 knee) was replaced by RT prosthesis (RT-PLUS(TM) Solution, PLUS). After operation, the valgus deformity of all patients was corrected and all patients could walk 100 m with or without the help of walking holders. The average range of motion (ROM) was improved from the pre operative. 95.6 degrees (85 degrees - 110 degrees ) to the post-operative 117.1 degrees (100 degrees - 125 degrees ). The average femorotibial angle (FTA) was corrected from the pre-operative. 27.6 degrees (20 degrees - 40 degrees ) to the post-operative 6.8 degrees (5 degrees - 9 degrees ). The Knee Score System (KSS) score and functional score were improved from the pre-operative 22.7 points (9 - 48 points) and 26.5 points (12 - 55 points) to the post-operative 86.4 points (85 - 95 points) and 89.1 points (80 - 95 points) respectively. Follow-up showed that the FTA remained unchanged and the knee stability of all patients was good. CONCLUSION: Through lateral approach, "Z" plasty of the capsule can release the lateral structure and decrease the pressure of common peroneal nerve. For TKR with moderate to severe fixed valgus knee, lateral approach is an effective way to correct the deformity. PMID- 17923011 TI - [The role of posterolateral structure and posterior cruciate ligament in external rotatory stability of the knee]. AB - OBJECTIVE: To investigate the changes of external rotation stability of the knee after the breaking and reconstruction of the posterolateral structure (PLS) of knee joint. METHODS: The femurs of 16 fresh cadaveric lower limbs were fixed to the base of rotating holders with the knee joints in full extension, or at an angle of 30 degrees , 60 degrees , or 90 degrees ; while the tibia was attached to a free rotary holder. The external rotation of the tibia was measured with a 5 kg x m x s(-2) tibial torque. The external rotation angle of the tibia was measured after the transectioning of the popliteus tendon (PT), popliteus muscle (PM), popliteofibular ligament (PFL), fibular collateral ligament (LCL), or popliteus tendon (PT). Autogenous hamstring tendon was used to reconstruct the PCL, and the Achilles tendon was used to reconstruct the PFL and PT, then the external rotation angle of the tibia was measured again. RESULTS: The isolated sectioning of PCL did not increase the tibial external rotation angle. When the knee was flexed the external rotation angle was 14.1 degrees in the intact knee and was 14.57 degrees in the PCL transectioning group (q = 0.47, P > 0.05). After the PFL was sectioned, when the knee joint was flexed at 60 degrees the external rotation angle was 16.94 degrees (q = 2.84, P < 0.05). After the PT was sectioned (PCL + PFL + PT), when the knee joint was flexed at 60 degrees the external rotation angle was 28.1 degrees (q = 14.01, P < 0.05). After isolated PCL reconstruction the external rotation angle was still bigger than that of the normal knee: for example, when the knee joint was at the angle of 60 degrees the external rotation angle was 27.67 degrees (q = 0.425, P < 0.05). There was a significant decrease in external rotation compared with PT section (P < 0.05). After the combined PCL + PLS reconstruction when the knee joint was at the angle of 60 degrees the external rotation angle of the knee was 14.51 degrees (q = 0.412, P < 0.05), however, the range of change was less than 1 degree. CONCLUSION: Isolated PCL section produces no change in external rotation. Complex injury of PCL and PSL can produce instability in external rotation. PT has the greatest important role to resist external rotation, then the greatest increase in external rotation can be found after PT is sectioned. Isolated PCL reconstruction can not completely restore the posterolateral stability of the knee. PCL reconstruction can partly restore the stability of the external tibial rotation. Only the combined PC + PLS reconstruction can reset the knee to physiological stability of external rotation. PMID- 17923012 TI - [Functional magnetic imaging of brain cortical activities in minimally conscious patients stimulated by visual presentation]. AB - OBJECTIVE: To investigate whether a large-scale visual network and visual consciousness are still retained in the patients with minimally conscious state (MCT) and the possibility of promoting the rehabilitation of brain function by visual stimulation. METHODS: Fifteen standardized pictures provided by the International Affective Picture System (IAPS2005) were passively presented to 10 normal adult controls and 9 patients in MCT recovering from severe craniocerebral injury or cerebral hemorrhage. Functional magnetic imaging was conducted. The AFNI software was used to process the images thus collected. RESULTS: The primary visual cortex, occipital lobe, and the secondary visual cortexes, including temporal lobe, parietal lobe, fusiform gyrus, orbital gyrus, and prefrontal lobe were all activated in the 10 normal persons. Similar visual activation was found in 2 of the patients in MCS, however, with a smaller activation volume. Activation of partial visual network was found in 3 patients and the other 4 failed to be activated. CONCLUSION: It is feasible to promote the rehabilitation of brain function by visual stimulation in the patients in MCS. PMID- 17923013 TI - [Expression of receptor-binding cancer antigen expressed on SiSo cells in endometrial carcinoma and the correlation thereof with the expression of estrogen receptor subtypes]. AB - OBJECTIVE: To investigate the expression of receptor-binding cancer antigen expressed on SiSo cells (RCAS1) and estrogen receptor subtypes in normal, hyperplastic, and carcinomatous endometrium and to explore their possible roles in carcinogenesis and progression of endometrial carcinoma. METHODS: Immunohistochemistry and semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) were applied to detect the protein and mRNA expression of RCAS1, estrogen receptor (ER) alpha and ERbeta in 20 specimens of normal endometrium, 24 specimens of hyperplastic endometrium, and 50 specimens of carcinomatous endometrium. Western blotting was used to detect the RCAS1 protein expression. RESULTS: Normal endometrium and simple and complex hyperplastic endometrium showed a P pattern of RCAS1 expression; carcinomatous endometrium showed the D pattern; and the atypical hyperplastic endometrium showed both, 30% (6/20) of which showed D pattern. The high level RCAS1 protein expression rates of the normal, simple and complex hyperplasia, atypical hyperplasia, and endometrial carcinoma were 0% (0/20), 9.1% (2/22), 45.0% (9/20), and 68.0% (34/50) respectively with a significant difference between any 2 groups (all P < 0.05). The high level expression of RCAS1 was detected more frequently in the carcinomatous endometrium with deep myometrial invasion, vascular invasion, and positive of ERalpha (all P < 0.05). The Western blotting and RT-PCR results were correlated with the immunohistochemistry results. The RCAS1 mRNA level was positively correlated with the ERalphamRNA level (P < 0.01), but not correlated with the ERbetamRNA level (P > 0.05). CONCLUSION: The expression and distribution of RCAS1 may be involved in the malignant transformation of endometrium, and RCAS1 coexpression with ERalpha may be associated with the development and metastasis of endometrial carcinoma. PMID- 17923014 TI - [Expressions of heme oxygenase-1 and apoptosis-modulating proteins in peri hematoma cortex after intracerebral hemorrhage in human being]. AB - OBJECTIVE: To observe the expression of heme oxygenase-1 (HO-1) and Bcl-2, an apoptosis-modulating protein, in the neurons surrounding the hematoma in human being. METHODS: Specimens of cerebral cortex tissue 1 - 3 cm around the hemorrhagic focus with the size of 2.0 cm x 1.5 cm x 0.3 cm were collected during autopsy from 39 patients, 17 males and 22 females, aged 62.8 (36 - 84), who died from intracerebral hemorrhage 2 - 10 h, 17 - 30 h, 36 - 96 h, 120 - 216 h, or 240 - 408 h before. Specimens of brain tissue of the same size at the opposite side were collected as controls. Immunohistochemistry was used to examine the expression of HO-1 and Bcl-2 protein. RESULTS: (1) Expression of HO-1 could be detected in the specimens of the 2 h group, increased in the specimens of the 2 - 10 h group [(5.1 +/- 2.0)/HP], reached the peak in the 17 - 30 h group [(11.3 +/- 0.9)/HP], then began to decrease in the specimens of the 240 - 408 h group [(6.4 +/- 0.6)/HP] (F = 42.80, P < 0.001). The HO-1 expression of the control group remained negative at any time-point. (2) Expression of Bcl-2 could be detected in the specimens of the 2 - 10 h group [(4.2 +/- 1.7)/HP], was increased in the 17 - 30 h group [(6.6 +/- 0.5)/HP], reached the peak in the 36 approximately 96 h group [(8.9 +/- 1.1)/HP], then began to decrease, and was (4.7 +/- 0.6)/HP in the 240 approximately 408 h group (F = 29.59, P < 0.001). The Bcl-2 expression remained negative at any time point in the control group. (3) The expressions of HO-1 was positively correlated with the expression of Bcl-2 (r = 0.66, P < 0.001). CONCLUSIONS: Over-expression of HO-1 and Bcl-2 in the neurons provide a potential protection or destruction mechanism after intracerebral hemorrhage in human. PMID- 17923015 TI - [Effect of hydroxyethyl starch 130/0.4 on S100B protein level and cerebral oxygen metabolism in open cardiac surgery under cardiopulmonary bypass]. AB - OBJECTIVE: To observe the effect of hydroxyethyl starch (HES) 130/0.4 on S100B protein level and cerebral metabolism of oxygen in open cardiac surgery under cardiopulmonary bypass (CPB) and to explore whether it has the protective effect of 6%HES130/0.4 as priming solution on cerebral injury during CPB and explore the probable mechanism. METHODS: Forty patients with atrioseptal defect or ventricular septal defect scheduled for elective surgical repair under CPB with moderate hypothermia were randomly divided into two equal groups: HES 130/0.4 group (HES group) in which HES 130/0.4 (voluven) was used as priming solution and gelatin group (GRL group) in which gelofusine (succinylated gelatin) was used as priming solution. ECG, heart rate (HR), blood pressure (BP), mean arterial pressure (MAP), central venous pressure (CVP), arterial partial pressure of oxygen (P(a)O(2),), arterial partial pressure of carbon dioxide (P(et)CO(2)) and body temperature (naso-pharyngeal and rectal) were continuously monitored during the operation. Blood samples were obtained from the central vein for determination of blood concentrations of S100B protein at the following time points: before CPB (T(0)), 20 minutes after the beginning of CPB (T(1)), immediately after the termination of CPB (T(2)), 60 minutes after the termination of CPB (T(3)), and 24 hours after the termination of CPB (T(4)). The serum S100B protein levels were measured by ELISA. At the same time points blood samples were obtained from the jugular vein and radial artery to undergo blood gas analysis and measurement of blood glucose, based on which the cerebral oxygen metabolic rate/cerebral metabolic rate of glucose (CMRO(2)/CMR(GLU)) was calculated. RESULTS: Compared with the time point of immediately before CPB (T(0)), The S100B protein level of the 2 groups began to increase since the time point T(1), peaked at the time point T(2), began to decrease gradually since the time point T(3), and were still significantly higher than those before CPB at the time point T(4) (all P < 0.01), and the S100B protein levels at different time points of the HES group were all significantly lower than those of the GEL group (all P < 0.01). The S(jv)O(2) and CMRO(2)/CMR(GLU) levels of both groups increased at the time point T(1), decreased at the time points T(2) and T(3), and then restored to normal at the time points T(4). In the GEL group there were no significant differences in the levels between any 2 different time points, however, in the HES group S(jv)O(2) and CMRO(2)/CMR(GLU) levels at T(1) was significantly higher than those at the other time points (P < 0.05 or P < 0.01). CONCLUSION: S100B protein increases significantly in open cardiac surgery under CPB. HES130/0.4 lowers the S100B protein levels from the beginning of CPB to one hour after the termination of CPB with the probable mechanism of improving the cerebral metabolism of oxygen. 6%HES130/0.4 as priming solution may play a protective role in reduction of cerebral injury during CPB and open cardiac surgery. PMID- 17923016 TI - [Repair of soft-tissue defect of amputation stumps of the forearm with free flap from the traumatic amputated extremity]. AB - OBJECTIVE: To report the clinical results of the repair of soft-tissue defect of amputation stumps of the forearm with free flap from the traumatic amputated extremity. METHODS: Five patients, 4 males and 1 female, aged 32 (22 - 43), with soft-tissue defect of the remaining stump of the traumatic amputation of the forearm, 3 cases in the right forearm and 2 cases in the left forearm, underwent repair of the defect by free flap from the traumatic amputated extremity with the size of the flaps ranging from 8 cm x 9 cm to 9 cm x 12 cm. The patients were followed up for 2.6 years (1.5 - 3.5 years). RESULTS: Superficial infection occurred in one patient postoperatively and the wound was gradually healed by daily wound dressings. All the flaps survived completely with satisfactory clinical results. The cosmetic appearance on the recipient area was good and the function of the elbow recovered satisfactorily. CONCLUSION: The flap from the traumatic amputated extremity has a constant vascular anatomy and a long vascular pedicle, so that dissection of the flap can be accomplished easily. The surgery allows to preserve the functional length or the elbow function of the remaining stump of the traumatic amputation. PMID- 17923017 TI - [Diagnosis and treatment for postoperative lobar torsion]. AB - OBJECTIVE: To analyze retrospectively 8 cases of postoperative lobar torsion after thoracotomy. METHODS: 8 cases of postoperative lobar torsion were collected (5 men and 3 women; median age, 55.0 +/- 7.7 years), including lobectomy 4 (left upper lobe of lung 2, right upper lobe of lung 2), esophageal carcinosectomy 2, resection of schwannoma in the right upper mediastinum 1, and descending aorta replacement 1. RESULTS: The postoperative lobar torsions were right middle lobe 2, right upper lobe 1, left upper lobe 3, left lower lobe 1, left lung 1. The median peak temperature was 38.4 degrees C (range, 37.8 - 40.2 degrees C) and the median white blood cell count was 10.6 x 10(9) cells/L (range, 9.3 - 14.9 x 10(9) cell/L) during the first 48 hours postoperatively. Postoperative radiographs demonstrated pulmonary infiltrates and volume loss in 6 patients and complete opacification in 2 patients. The diagnosis of lobar torsion was made a median of 4 days (range, 2 - 14 days) after the initial operation; 6 patients underwent resection of lung and recovered; 2 had the injured lobe or lung rotated and died. Complications after reoperation included respiratory failure in 2 patients, atrial arrhythmia in 2 patients. Median hospitalization was 24 days and range from 10 to 56 days. CONCLUSIONS: The mobilization of hilus of lung or residual pulmonary atelectasis is the main mechanism of the lobar torsion after thoracotomy. Lobar torsion represents a difficult diagnostic dilemma in the early postoperative period after thoracotomy. Exploratory thoracotomy must be performed without delay. The injured parenchyma should be sacrificed unless the diagnosis is obtained very early. When the injured lobe or lung is rotated back into normal position, simultaneous endotracheal suction is very important to prevent aspiration of fluid from the obstructed part of the bronchial tree to the uninvolved segments and dangerous postoperative hypoxia. PMID- 17923018 TI - [Effect of Smad7 on transforming growth factor-beta1-induced alveolar epithelial to mesenchymal transition]. AB - OBJECTIVE: To investigate whether transforming growth factor beta1 (TGF-beta1) can induce in vitro alveolar epithelial-mesenchymal transition (EMT), and whether Smad7 gene transfer can block this transition and the possible signaling mechanism. METHODS: Rat alveolar type II epithelial cells of the line RLE-6TN were cultured. TGF beta1 (3 ng/mL) was added into the culture fluid. Lipofectamine 2000 was used to transfect Smad7 gene to the RLE-6TN cells. The expression of the markers of the epithelial cells, including E-cadherin and cytokeratin-19 (CK19), and markers of mesenchymal cells, including fibronectin (FN), vimentin, and alpha-smooth muscle actin (alpha-SMA) were assayed using Western blotting and real-time PCR. The morphological alterations were examined by phase-contrast microscope while the ultrastructure changes were examined by electron microscope. RESULTS: Smad7 was successfully transfected to the RLE-6TN cells. Before transfection TGF-beta1 treatment could lead to the expression upregulation of the mesenchymal markers and downregulation of the epithelial markers at the levels of both mRNA and protein, and after transfection, the mesenchymal makers were downregulated while the epithelial markers were upregulated. Before the transfection, TGF-beta1 treatment could lead to the expression upregulation of phosphorylated Smad2/3 which did not obvious change following transfection. TGF beta1 treatment could induce the EMT process of the RLE-6TN cells. Smad7 gene transfected into the RLE-6TN cells could block the process of EMT. Exposed of the RLE-6TN cells to TGF beta1 resulted in degeneration, tumefaction, and gradual disappearance of the osmiophilic multilamellar bodies, markers of type II alveolar epithelial cells. CONCLUSION: Under TGF beta1 treatment, RLE-6TN undergoes a conversion process into myofibroblasts in vitro with the conversion mechanism related to Smad signaling pathway, and transfection of Smad7 gene can partly reverse this process. PMID- 17923020 TI - [Impact of different drugs and different volumes injected intraspinally on the nerve function: experiment with rats]. AB - OBJECTIVE: To investigate the impact of different drugs and different volumes injected intraspinally on the nerve function and find out the safe intraspinally injection volume. METHODS: Ninety-six adult Sprague-Dawley rats were randomly divided into 12 equal groups to be injected intraspinally with normal saline (NS) 2 microl, 4 microl, 6 microl, or 10 microl, viral buffer 2 microl, 4 microl, 6 microl, or 10 microl, or Ad-LacZ (1 x 10(9) pfu/ml) 2 microl, 4 microl, 6 microl, or 10 microl at the level of T13 respectively immediately after the laminectomy was performed. Another 6 adult SD rats only underwent T13 luminal decompression and used as controls. Basso-Beattie-Bresnahan locomotor rating scale was used to evaluate the motor function of their hind limbs after 1, 2, 3, 4, 5, and 6 days, and 1, 2, 3, and 4 weeks. One and 2 weeks after the injection 2 rats from each group were killed and the remaining 4 rats in each group were killed 4 weeks after the injection. Histological examination was done with HE staining and toluidine blue staining. X-gal staining was used to observe the expression of adenovirus in the spinal cord. RESULTS: The extent of motor deficits and tissue damage increased following the increase of intraspinal injection volume (P = 0.000). The severity of damage of the Ad-LacZ was significantly greater than those of the NS and virus buffer groups (P = 0.044), however, there were no significant differences in the severity of damage among NS groups and virus buffer groups (P = 1.000). X-Gal staining showed that the reporter gene LacZ was expressed in all Ad-LacZ groups. The great the dose the more severe the damage. Four weeks later the effective residual rates f the grey and white matters of the Ad-LacZ groups were all significantly lower than those of the other 2 groups, however, there were no significant differences in the effective residual rates f the grey and white matters among the NS and virus buffer groups. CONCLUSION: Intraspinal injection results in functional deficit and tissue damage in the spinal cord. The extent of tissue damage increases following the increase of intraspinal injection volume. Different drugs cause different tissue damage. The intraspinal injection of Ad-LacZ results in more tissue damage. The volume of 4 microl is the safe volume for injection, 6 microl is a critical volume. PMID- 17923021 TI - [Potential of chondrogenesis of bone marrow stromal cells co-cultured with chondrocytes on biodegradable scaffold: in vivo experiment with pigs and mice]. AB - OBJECTIVE: To explore the feasibility of in vivo chondrogenesis of bone marrow stromal cells (BMSCs) co-cultured with chondrocytes on biodegradable scaffold. METHODS: Porcine BMSCs were isolated, expanded and labeled with enhanced green fluorescent protein (EGFP), and then were mixed with articular chondrocytes isolated from porcine knee joint at the ratio of 1:1. The mixed cells were seeded onto polyglycolic acid (PGA) scaffold at the ultimate concentration of 5.0 x 10(7)/ml (co-culture group). Pure chondrocytes and BMSCs of the same ultimate concentration were seeded respectively onto the scaffold as positive control group and negative control group. After two weeks' culture in vitro, they were planted subcutaneously into nude mice respectively. These specimens were collected after in vivo implantation for 8 weeks to undergo microscopy. Laser confocal microscopy was used to observe the distribution of EGFP-labeled cells in the tissue. RT-PCR was used to examine the expression of collagen type II and aggrecan. Immunohistochemistry was used to observe the protein expression of collagen type II. RESULTS: The cell-scaffold constructs of the co-culture group and positive control group, could maintain the original size and shape no matter in vitro or in vivo. After 8 weeks' in vivo implantation, the constructs in both co-culture group and positive control group formed cartilage-like tissue with typical histological structure and extracellular matrix staining similar to those of the normal cartilage. The GAG content and compressive modulus of the co culture group reached over 80% of those of the positive control group. Confocal microscopy revealed the presence of EGFP-labeled cells in the engineered cartilage lacuna. Histological examination showed that the constructs of the negative control group shrunk gradually after in vivo implantation with no typical cartilage-like tissue formation. CONCLUSION: In vitro co-cultured BMSC chondrocyte-PGA constructs have the potential to form mature cartilage-like tissue in subcutaneous non-chondrogenesis environment, indicating that chondrocytes still provide enough signals for BMSC chondrogenic differentiation. PMID- 17923026 TI - [The suggestion of the experience with prevention and therapy of colorectal cancer in the industrialized country]. PMID- 17923022 TI - [Relationship between survivin expression and chemosensitivity of human lung cancer cells]. AB - OBJECTIVE: To investigate the relationship between survivin expression and the chemosensitivity in multiple drug resistant human lung cancer cell line METHODS: Human lung cancer cells of the parental cell line H460 and multiple drug resistant cell line H460/cDDP were cultured respectively. RT-PCR and Western blotting were used to detect the mRNA and protein expression of survivin. siRNA targeting survivin was transfected into H460/cDDP cells by Liposomes approach. The expression change of survivin protein after transfection was examined by Western blotting. Cisplatin (DDP) and paclitaxel of different concentrations were added into the culture fluid of the H460/cDDP cells respectively. Methyl thiazolyl tetrazolium (MTT) assay was used to calculate the IC(50) value so as to detect the cytotoxicity of cisplatin and paclitaxel to the cells. RESULTS: The mRNA expression and protein expression of survivin in the H460/cDDP cells were both significantly higher than those in the H460 cells. 24 h, 48 h, and 72 h after transfection of the survivin siRNA the survivin protein expression levels of the H460/cDDP were decreased by 67.0%, 76.2%, and 92.3% respectively. MTT assay showed that the cisplatin IC(50) levels of the H460/cDDP and H460 cells were 6.3 +/- 0.6 mg/L and 0.8 +/- 0.1 mg/L respectively, and the paclitaxel IC(50) levels were 12.7 +/- 1.2 mg/L and 1.5 +/- 0.3 mg/L respectively. 48 h after the transfection of survivin siRNA, the cisplatin and paclitaxel IC(50) levels of the H460/cDDP cells were decreased to 4.0 +/- 0.9 mg/L (P = 0.002) and 8.1 +/- 1.7 mg/L (P = 0.003) respectively. CONCLUSION: The survivin expression of the resistant cell line H460/cDDP was up-regulated compared with that of the parental cell line H460. Survivin siRNA sensitized the H460/cDDP cells to both cisplatin and paclitaxel. Survivin participates in the multiple drug resistance mechanism of human lung cancer cells. PMID- 17923028 TI - [Strategy study on colorectal cancer liver metastasis]. PMID- 17923029 TI - [Epidemiology investigation of colorectal cancer on community group in Guangdong province]. AB - OBJECTIVE: To explore the epidemiology characteristics of colorectal cancer on community group in Guangdong province. METHODS: Huidong was took as the research spot to investigate the sickness and incidence of colorectal cancer among a whole community group in Guangdong province, and two projects were used simultaneously: "screening of colorectal cancer in high risk group" and "screening project using fecal occult blood test". The numeration time for population of 10 towns was set at July 1, 2005. Five towns were sampled out from Huidong with 100,859 persons. Meanwhile, the diseases and death cause registering system was established. RESULTS: The incidence of colorectal cancer in Huidong community group was 15.2/100,000 (male 17.2/100,000, female 13.0/100,000), the age-adjusted incidence was 17.6/100,000 (male 19.5/100,000, female 16.7/100,000). The morbidity was 41.9/100,000 (male 46.5/100,000, female 37.2/100,000), the age-adjusted morbidity was 49.0/100,000 (male 54.1/100,000, female 45.6/100,000). The mortality was 5.0/100,000 (male 5.9/100,000, female 4.0/100,000) and the age-adjusted morbidity was 6.4/100,000 (male 7.2/100,000, female 5.7/100,000). CONCLUSION: The incidence of colorectal cancer in Huidong community group is lower than that of high incidence area in China, but gets close to the normal incidence area of China and general level of the world. PMID- 17923031 TI - [The role of secreted Wnt-antagonist genes hypermethylation in early detection of colorectal tumor]. AB - OBJECTIVE: To investigate the functions of promoter hypermethylation of secreted Wnt-antagonist genes in colorectal tumorigenesis and progression. METHODS: Two colorectal cancer cell lines, HCT116 and SW480, were treated by 5-aza-2' deoxycytidine (DAC) and trichostatin A (TSA) for demethylation. The promoter hypermethylation and expression of sFRP and WIF-1 genes in different stages of colorectal tumor and colorectal cancer cell lines were detected by methylation specific PCR and reverse transcription PCR, respectively. RESULTS: None of the normal colorectal mucosa samples showed methylated bands of any sFRP and WIF 1genes. Hypermethylation of sFRP1, 2, 4, 5 and WIF-1 was detected in 93.1% (67/72), 83.3% (60/72), 36.1% (26/72), 52.8% (38/72) and 84.7% (61/72) of adenocarcinomas, 87.9% (29/33), 81.8% (27/33), 24.2% (8/33), 57.6% (19/33) and 72.7% (24/33) of adenomas, 52.6%, 28.9%, 2.6%, 18.4%, 23.7% of the adjacent normal mucosa. Methylation was more frequently found in colorectal tumors than in normal mucosa and adjacent normal mucosa from patients with tumor (P < 0.05). No significant association between Wnt-antagonist genes hypermethylation and clinicopathological characteristics was found (P > 0.05). SFRP1, 2, 4, 5 and WIF 1 genes were methylated in HCT116 cell line. SFRP1, 2 and WIF-1 were methylated in SW480 cell line. The mRNA expression of sFRPs and WIF-1 genes was absent or significantly downregulated (P < 0.01) when they were methylated in two colorectal cancer cell lines. SFRP3 was expressed in two colorectal carcinoma cell lines. DAC/TSA combination treatment re-expressed the silenced sFRPs and WIF 1 genes mRNA expressions effectively. A single application of TSA could not re express sFRPs and WIF-1 genes mRNA expressions. The influence of demethylation treatment on sFRP3 expression was minimal. CONCLUSION: Hypermethylation of Wnt antagonist genes is a common early event in the evolution of colorectal tumor. Methylation of sFRP1, 2, 5 and WIF-1 genes might serve as biomarkers for the early detection of colorectal tumor. PMID- 17923032 TI - [Selective cyclooxygenase-2 inhibitors for the prevention of colorectal adenomas: a meta-analysis]. AB - OBJECTIVE: To determine the validity and safety of selective cyclooxygenase (COX) 2 inhibitors for the prevention of colorectal adenomas. METHODS: The relevant data were retrieved from Medline (1966 to 2006), OVID (1996 to January 2007), EMBASE (1980 to January 2007), Chinese Cochrane Centre databases (up to January 2007) and Chinese Biological Medicine Disk (CBM disk, 1997 to 2007). Methodological quality assessment was based on the Cochrane Reviewers' Handbook and Jadad's Score Scale. Statistical software RevMan4.2 was used for meta analysis. RESULTS: Six randomized clinical trials (5708 patients) were included in the study. Compared with placebo, selective COX-2 inhibitors lowered the detection rates of both adenomas and advanced adenomas (RR: 0.70, 95% CI: 0.55 - 0.88, P = 0.0003 and RR: 0.69, 95% CI: 0.53 - 0.89, P = 0.005). No significant difference was observed in the number of adverse events between patients taking selective COX-2 inhibitors and those taking placebo (RR: 1.07, 95% CI: 0.98 - 1.17, P = 0.11). Compared with placebo, selective COX-2 inhibitors increased the risk of cardiovascular events (RR: 1.21, 95% CI: 1.09 - 1.33, P = 0.0002). CONCLUSION: Selective COX-2 inhibitors can induce sufficient regression of colorectal adenomatous polyps and thus be used for chemoprevention of colorectal neoplasms. However, because of the potential cardiovascular events, it is not routinely recommended for this indication. PMID- 17923033 TI - [DelD631: a novel mutation of the RET proto-oncogene in multiple endocrine neoplasia type 2A (MEN2A)]. AB - OBJECTIVE: To detect RET mutations in a rare Chinese big family with Multiple endocrine neoplasia type 2A (MEN2A). METHODS: One MEN2A family, including the proband, have 22 members of two generations, it is a rare big family in modern Chinese families. The DNAs of the 22 members from the family including 4 patients were extracted from blood leukocytes, PCR and gene sequencing of PCR products by an automated DNA sequencer were applied to scan the exon10 and 11 of RET proto oncogene. Sequencing results were compared with the Pubmed's. Clone sequencing was adopt to further confirm the results, then verifying the novel mutation through the human gene mutation database at the institute of medical genetics in cardiff. Invitrogen biotechnology company (Shanghai) provided the technology of clone sequencing. RESULTS: A novel deletion mutation of D631 (GAC) (del D631) was detected in exon11 of the RET proto-oncogene in 4 MEN2A patients of the family, this rare deletion mutation of D631 (GAC) lead base sequence of TGC(angle)GACGAGCTG change to TGCGAGCTG. Besides 4 MEN2A patients, the son of II6 (the first class relative) was found to be a carrier of delD631 mutation. CONCLUSION: A novel deletion mutation (del D631) of RET proto-oncogene was detected in the family with MEN2A and it has never been reported before in the world. DelD631 may be related to the late onset of MEN2A compared to the cysteine mutations and pheochromocytoma might be the first manifestation prior to the development of MTC. PMID- 17923034 TI - [Study on gag-pol region characteristics of HIV-1 strains epidemic in Liaoning province]. AB - OBJECTIVE: To investigate the genetic subtypes and sequence characteristics of gag-pol region of HIV-1 epidemic in Liaoning province. METHODS: 99 whole blood samples of HIV-1 infected individuals of Liaoning province were collected. Provirus DNA was extracted from anti-coagulated peripheral whole blood, and then the sequences of gag-pol region were amplified through nested PCR and then sequenced. Genetic subtypes were identified by phylogenetic analysis, controlled with new version subtype reference sequences of HIV sequence database. The genetic distances of different coding regions were calculated by MEGA2.0 with Kimura 2-parameter model. The synonymous (Ks) and anonymous (Ka) as well as the ratio of Ks/Ka were analyzed by SNAP tool in HIV sequence database. RESULTS: There are many HIV-1subtypes epidemic in Liaoning province. We found five subtype: A, Thai B, B, C and G, five CRFs: CRF01, CRF03, CRF06, CRF07, CRF08, and one inter CRF recombinant ICR07/08 in 99 HIV-1 infected individuals of Liaoning province. Thai B subtype was the main subtype epidemic in Liaoning, next were CRF01_AE. In gag-pol region, the genetic distances of p17, p2-p6 were higher than p24, protease and RT coding region. The genetic distance of p24 region in B subtype strains were relative high compared with other subtypes. The genetic distance and the value of Ks and Ka were rather low in each region of CRF07 and CRF08. The ratio of Ks/Ka of p2-p6 region was the lowest in each subtype, the mean value was below 1. The ratio of Ks/Ka of RT region was as high as 10.45. CONCLUSION: The HIV-1 genetic subtypes in Liaoning province were rather complex. The p24, protease and RT coding region coding region were more conserved in gag pol region. P24 region of B subtype strains received more positive selective pressure, while the RT region of CRF01_AE strains received more negative selective pressure. CRF07_BC and CRF08_BC were the youngest strains transmitted to Liaoning province and showed strong founder effect. PMID- 17923036 TI - [The relationship between adiponectin and coronary heart disease metabolic risk factors in non-diabetic male patients]. AB - OBJECTIVE: To investigate the correlation between adiponectin and metabolic risk factors of coronary heart disease in non-diabetic male patients. METHODS: 215 non diabetic males aged (59 +/- 9) underwent coronary angiography (CAG) and then divided into coronary heart disease group (n = 127) and control group (n = 88) according to the CAG results. The anthroposomatologic parameters were measured. Blood sugar and lipids were examined. Adiponectin levels was detected with radioimmunoassay. RESULTS: The mean adiponectin level of the coronary heart disease group was 7.9 (5.7 - 11.6) mg/L, significantly lower than that of the control group [10.0 (7.0 - 13.5) mg/L, P = 0.014]. The adiponectin level of females was lower than those of the males, no matter whether with coronary heart disease. Partial correlation analysis showed that the adiponectin level was negatively correlated with the serum triglyceride, total cholesterol, post challenge glucose level and body mass index (r = -0.250, -0.169, -0.167, -0.246, and -0.198, all P < 0.05), and positively correlated with HDL-C (r = 0.201, P = 0.001) after the age factor was controlled. Logistic multivariable stepwise regression analysis demonstrated levels of triglyceride and adiponectin independently associated with the presence of coronary heart disease (OR = 2.277 and 0.533 respectively). CONCLUSION: Adiponectin is influenced by metabolic disorders. Higher adiponectin level is a protective factor of coronary heart disease. PMID- 17923037 TI - [Evaluation of 126 radical cystectomy with orthotopic ileal or sigmoidocolic neobladder]. AB - OBJECTIVE: To evaluate the long-term clinical effects of orthotopic ileal or sigmoidocolic neobladder. METHODS: One hundred and twenty six patients with bladder cancer who underwent radical cystectomy and orthotopic ileal or sigmoidocolic neobladder from 1989 to 2001 were followed up, the clinical data was collected and analysed. Hautmann orthotopic ileal neobladder was performed on 84 cases and orthotopic sigmoidocolic neobladder was performed on 42 cases; Lymph node clearing during surgery was performed on 62 cases, chemotherapy and radiotherapy was performed on 64 cases after surgery. The continence and complications were compared between sigmoidocolic group and ileal group, the tumor recurrent rate and the 5-year survival rate were compared between lymphnode clearing group and chemoradical therapy group. RESULTS: Complete follow up was performed in 122 cases. Ureter broaden and urine backflow rate were higher in sigmoidocolic group than in ileal group (P < 0.05), nocturnal continence rate in sigmoidocolic group was higher than in ileal group (P < 0.05); Post-surgical tumor recurrent rate in lymphnode clearing group was lower than in chemoradical therapy group (P < 0.05), the 5-year survival rate in lymphnode clearing group was higher than in chemoradical therapy group (P < 0.05). The overall short-term complication rate was 15.9% (20/126), the overall long-term complication rate was 9.8% (12/122). CONCLUSION: The effects of orthotopic ileal or sigmoidocolic neobladder were satisfactory with low complication rate, lymphnode clearing during the surgery can increase the 5-year survival rate when compared with the chemoradical group. PMID- 17923038 TI - [Relevance of posttransplant HLA-II antibodies production on long-term survival of renal allograft]. AB - OBJECTIVE: To evaluate the clinic relevance of anti-HLA-II antibodies on allograft long-term survival. METHODS: Perioperative sera of 118 cadaveric kidney recipients were tested by ELISA for anti-HLA-II antibodies in our prospective cohort study. All recipients who divided into different groups according to HLA antibody production were followed-up. RESULTS: (1) Anti-HLA-II antibody-positive recipients were associated with significantly lower graft survival (78.6% vs 84.4%; 71.4% vs 80.0%; P = 0.002) and death-censored graft survival (85.7% vs 92.2%; 82.1% vs 90.0%; P = 0.003) at 3 and 4 years compared to antibody-negative recipients. (2) Anti-HLA-II antibody-positive recipients were associated with a significantly increased risk for decline in renal function at 3 and 4 years (39.3% vs 33.3%; 46.4% vs 38.9%, P = 0.001). (3) There was statistically non significance difference in late-acute rejection rate between two groups (10.7% vs 13.3%, P > 0.05). CONCLUSION: Posttransplant HLA-II antibodies perhaps are one of the most important influential facts on allograft long-term survival and could be used to predict the prognosis of allograft. PMID- 17923039 TI - [JAK3 mRNA level in graft versus host disease patients: a preliminary study]. AB - OBJECTIVE: To quantify peripheral blood mononuclear cell JAK3 mRNA levels after allogeneic haematopoietic stem cell (allo-HSCT) transplantation, and to explore its relationship with graft vs host disease (GVHD). METHODS: Serial monitoring of JAK3 mRNA levels by fluorescent quantitative reverse transcriptase polymerase chain reaction (FQ-RT-PCR) technique was performed on 26 cases (83 samples) after allo-HSCT. RESULTS: The mean JAK3/ABL expression levels was 8.71 in 14 patients without GVHD; the JAK3/ABL expression level was 31.94 in one patient with acute GVHD (aGVHD); the mean JAK3/ABL expression levels was 19.23 in 11 patients with chronic GVHD (cGVHD), and the cGVHD was diagnosed at the mean time of six months (3 - 12 months). The JAK3/ABL expression levels did not change in 4 patients one month after allo-HSCT, however, the levels increased again when GVHD were diagnosed in these 4 patients, and the JAK3/ABL expression levels decreased again when GVHD was remitted. CONCLUSION: The JAK3 expression level related to the remission and relapse in patients with GVHD. PMID- 17923040 TI - [Analysis of the correlation between PreS1-Ag, PreS2-Ag, HBeAg and HBV DNA in the serum of chronic hepatitis B patients]. AB - OBJECTIVE: To investigate the correlation between the serum indices and HBV DNA. METHODS: 100 chronic HBV patients and 40 healthy controls were enrolled in this study. The patients have not received any treatment with interferon (IFN) and similar nucleotide, and featured with normal ALT/AST, positive HBV DNA examined by using internal-quantitative standard PCR. Serum indices were measured by micro particle enzyme immunoassay analysis and ELISA. RESULTS: In chronic HBV patients with positive HBV DNA, the positive percentage were 75% for HBeAg, 25% for anti HBe, 69% for PreS1-Ag, and 77% for PreS2-Ag. In 75 HBeAg positive cases, there were 54 cases of PreS1-Ag positive and 60 cases of PreS2-Ag positive; while among anti-HBe positive cases, there were 14 cases of PreS1-Ag positive and 17 cases of PreS2-Ag positive. There was no significant difference of the positive percentage of PreS1-Ag and PreS2-Ag between HBeAg positive and anti-HBe positive (both P > 0.05). Under circumstance of HBeAg positive, the percentage of both PreS1-Ag and PreS2-Ag negative (8%) was significant lower than that of anti-HBe positive (28%) (P < 0.05). There were 72 cases with consistent positive or negative for both PreS1-Ag and PreS2-Ag. CONCLUSION: For hepatitis B patients with positive HBV DNA, the differences of positive percentages for HBeAg, PreS1-Ag and PreS2-Ag are moderate. The positive percentages of PreS1-Ag and PreS2-Ag do not correlate with HBeAg positive. The negative percentages of PreS1-Ag and PreS2-Ag correlate with anti-HBe positive. PMID- 17923042 TI - [Enhanced transduction efficiency of adeno-associated virus on cancer cells with the help of low dose adenovirus]. AB - OBJECTIVE: To investigate whether adeno-associated virus (AAV) could enhance its infection efficiency on cancer cells when combined with non-replicable adenovirus (Ad-null) in vitro as well as in vivo and to study its underlying mechanisms. METHODS: AAV2 particle was added into NCI-H460 tumor cell lines alone or in combination with different amount of adenovirus. 1 to 7 days after transduction, cells were observed and recorded with fluorescence microscope, the expression levels of report gene EGFP in tumor cells were examined by using flow cytometry and Western blotting, the expression of report genes luciferase was analyzed with luminometer to obtain the relative light units. After the establishment of tumor model, the nude mouse were administrated with AAV2 or AAV2 + Ad-null in tumors, and then tested their infection efficiency and expression levels with roperscientific bioluminescence tumor imaging system. RESULTS: The results obtained with the help of flow cytometry and luciferase assay suggested that the infection efficiency of AAV2 was enhanced significantly when combined with low dose Ad-null in vitro, the infection efficiency of AAV2 alone was 6.4% and it reached 55.2% when combined with 10 MOI Ad-null, Western blotting assay demonstrated that the protein expression level of reporter gene in tumor cells enhanced when combined with 10 MOI Ad-null compared with AAV2 infection alone, and the enhancement of reporter gene expression was observed in a concentration dependent manner; real-time PCR analysis confirmed that Ad-null enhanced the mRNA level of AAV2-EGFP but not the copies of genomic DNA of AAV2-EGFP. Ad-null significantly augmented the infection efficiency when tested on NCI-H460 tumor model. With the help of Ad-null, the signal of luciferin in nude mouse was 4.5 times more than that of control. CONCLUSION: The infection efficiency of AAV was enhanced significantly when combined with low dose Ad-null in vitro and in vivo, and it offers basis for further study of gene therapy by AAV. PMID- 17923043 TI - [Protein kinase C-betaI, betaII in mouse diabetic nephropathy kidney and its relation to nephroprotective actions of the angiotensin receptor blocker telmisartan]. AB - OBJECTIVE: To investigate the localization and expression of protein kinase C (PKC)-betaI, betaII in diabetic nephropathy (DN) mouse kidney and its relation to angiotensin receptor blocker telmisartan (Micardis). METHODS: Eighteen mice were divided into three groups: normal group, DN group and Micardis-treated group (n = 6, each group). The expression of PKC-betaI, betaII, transforming growth factor- beta 1 (TGF-beta1) and vascular endothelial growth factor (VEGF) in glomeruli was measured by semiquantitative immunofluorescence histochemistry, the localization of PKC-betaI, betaII was detected by confocal immunofluorescence laser scanning microscopy and the expression of PKC-betaI, betaII in renal cortex, outer and inner medulla were evaluated by semiquantitative Western blotting. RESULTS: Compared to normal mice, the expression of PKC-betaI and betaII on apical membrane of proximal tubule epithelial cells of DN mice was significantly increased, whereas the expression of PKC-betaII on cortical and inner medullary collecting duct was decreased. Western blotting detected increasing expression of PKC-betaI in the renal cortex and outer medulla (P < 0.01), and decreasing expression of PKC-betaII in renal cortex of DN mice (P < 0.01). Enhanced expression of PKC-betaI as well as TGF-beta1 and VEGF (P < 0.01) were shown in the glomeruli of DN mice, where the expression of PKC-betaII was decreased (P < 0.05). Meanwhile, PKC-betaI exhibited a positive correlation to TGF-beta1 (r = 0.649, P = 0.030), but no correlation to VEGF (r = 0.387, P = 0.079). Micardis could partly attenuate above changes. CONCLUSION: The localization and expression of PKC-betaI, betaII are altered in DN mice, PKC-betaI, betaII may change the function of proximal tubule and PKC-betaI may contribute to glomerular hypertrophy through influencing the expression of glomerular TGF-beta1. Treatment with Micardis can partly improve the abnormal expression and distribution of PKC betaI, betaII in kidneys of DN mice, which suggests that renin-angiotensin-system is implicated in the pathogenesis of DN by regulating the expression and activation of PKC-betaI, betaII isoforms. PMID- 17923044 TI - [Empirical study of the transportation of pulmonary surfactant-super oxide dismutase liposome to lung tissue in rats]. AB - OBJECTIVE: To study the feasibility of the transportation of pulmonary surfactant super oxide dismutase (PS-SOD) liposome to lung tissue in rats. METHODS: 32 Wistar rats were randomly divided into 4 groups (8 rats in each group): normal saline group, PS group, SOD group, PS-SOD liposome group. Each group was further divided into two groups (4 rats in each group), and the rats were respectively killed 2 and 24 hours after the operation. While the biological activity of SOD in irrigating solution and tissue homogenate were detected, lung tissue were labeled with fluorescent and then observed under microscope and transmission electron microscope. RESULTS: PS-SOD liposome was corps rounds with monolayer lipid with stable surface tension and antioxidative activity. At the point of 2 hours after operation, while the SOD biological activity of irrigating solution in PS-SOD liposome group (32.87 +/- 5.47) and SOD group (33.14 +/- 5.61) were obviously higher than that in normal saline group (2.15 +/- 0.17, P < 0.01), there was no difference between them (P > 0.05). The mean fluorescence optical density in PS-SOD liposome group (0.109 +/- 0.018) was lower than that in normal saline group (0.144 +/- 0.052) and PS group (0.143 +/- 0.026, P < 0.01). 24 hours after operation, the SOD biological activity of irrigating solution in PS-SOD liposome group (11.54 +/- 1.42) was the highest (P < 0.01) and the mean fluorescence optical density in PS-SOD liposome group (0.112 +/- 0.018) was the lowest (P < 0.01). The SOD biological activity of tissue homogenate in PS-SOD liposome group (2 h: 16.83 +/- 2.69, 24 h: 15.70 +/- 2.75) was higher than that in normal saline group (2 h: 5.79 +/- 0.93, 24 h: 5.84 +/- 1.31) and in SOD group (2 h: 7.07 +/- 1.04, 24 h: 6.11 +/- 1.06, P < 0.01) both at the point of 2 and 24 hours after the operation. Lots of PS-SOD liposome was observed in type II alveolar epithelial cells under transmission electron microscope. CONCLUSION: Intrathecal administ ration of PS-SOD liposome enhanced the transportation of SOD into lung tissue and its antioxidative activity. PMID- 17923045 TI - [The tone of sympathetic and expression of beta1 receptor in the dogs with sustained atrial fibrillation]. AB - OBJECTIVE: To detect the influence of sustained atrial fibrillation on autonomic nerve. METHODS: 7 healthy mongrel dogs were paced at 600/min. ter sustained reversing into sinus rhythm. HRV was obtained by Holter. Noradrenaline in left atrial appendage, left atrial, atrial septal, right atrial appendage and right atrial was detected. Nerve growth factor was determined and beta1 receptor in atrial myocardium was evaluated. All the indexes were compared with control animals. RESULTS: ter sustained, the tone of sympathetic increased. LF in dogs with long term (1682 +/- 362) ms2 and short term (1247 +/- 219) ms2 were higher than control dogs (798 +/- 154) ms2 (P < 0.01). The HF in dogs with long term (232 +/- 75) ms2 and short term (310 +/- 165) ms2 was lower than control dogs (1041 +/- 195) ms2 (P < 0.01). The SDNN [long term (32 +/- 7) ms, short term (51 +/- 7) ms] and HRV indexes (long term 16 +/- 4, short term 24 +/- 8) in group were inferior to corresponding indexes in control group [(103 +/- 14) ms and 38 +/- 5 respectively, P < 0.01]. The content of noradrenaline in myocardium was higher in group (P < 0.01). Furthermore, the increase of noradrenaline in the atrium was heterogeneous (P < 0.01), which was not observed in control group. The expression of NGF increased in group. beta1 receptor down regulated in group. CONCLUSION: Sustained caused increased tone of sympathetic, heterogeneous accumulation of noradrenaline and down regulation of beta1 receptor, which was mediated by NGF. PMID- 17923046 TI - [The clinical value of the 99Tcm-MIBI imaging in predicting the prognosis of malignant lymphoma]. AB - OBJECTIVE: To predict the chemotherapy response in Malignant lymphoma (ML) using 99Tcm-MIBI imaging, and to evaluate whether 99Tcm-MIBI scintigraphy parameters may have a precise predictive value in the expression of MDR1 and multidrug resistance-related-protein genes. METHODS: Twenty-three patients with histologically proved Malignant lymphoma underwent 99Tcm-MIBI scintigraphy before chemotherapy. Tumor-to-background [corrected] (T/B) ratios of both early (10 min) and late images (1 h) and the percentage rate of washout (WR%) were measured; The mRNA expressions of MDR1 and MRP were measured by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR); therapeutic reaction was evaluated by clinical and radiologic methods after completing 6 - 8 cycle chemotherapy with/without involved field radiotherapy for large tumors. RESULTS: The WR% (16 +/- 6) of both MDR1 and MRP simultaneously negative expression group was significance lower than both simultaneously positive expression group (33 +/- 5), and also lower than either MDR1 or MRP positive expression group (28 +/- 6) (both P < 0.01); There was no significant difference between the both simultaneously positive expression group and either MDR1 or MRP positive expression group (P = 0.26). The early images, late images T/B ratios and WR% of MDR1 positive group were 3.0 +/- 1.1, 2.5 +/- 0.8 and 17 +/- 7 respectively; MDR1 negative group were 3.4 +/- 1.0, 2.3 +/- 0.7 and 32 +/- 6 respectively. There were no significant difference between the MDR1 positive group and MDR1 negative group in either early images or late images T/B ratios (P > 0.05), but the WR% was significant different between them (P < 0.01). The early images, late images T/B ratios and WR% of MRP positive group were 3.1 +/- 1.2, 2.5 +/- 0.8 and 19 +/- 8 respectively; MRP negative group were 3.3 +/- 1.0, 2.3 +/- 0.7 and 31 +/- 6 respectively. The WR% of MRP positive group was significantly higher than that of MRP negative group (P = 0.003); T/B ratios of both early and late images were all significantly different between them (P = 0.72, P = 0.60). Both levels of MDR1 and WR% were significantly positively correlated with therapeutic response (both P < 0.05), but there was no significant correlation between levels of MRP and therapeutic response (P = 0.052). CONCLUSION: As a untraumatic imagology instrument, 99Tcm-MIBI can accurately reflect the expression and functional status of MDR1 and MRP, and can predict the therapeutic response of ML, thus could be used for individualized treatment planning. 99Tcm-MIBI would be benefit to ML patients. PMID- 17923047 TI - [A preliminary study of endoscopic transoral trans-posteriorwall pharynx resection of hypo-clivus chordoma]. AB - OBJECTIVE: To introduce a endoscopic surgical approach for hypo-clivus chordoma, and to explore the clinical value of the endoscopic resection of hypo-clivus chordoma. METHODS: Three hypo-clivus chordoma were resected by endoscopic transoral tans-posteriorwall pharynx approach. RESULTS: The MR image showed that total removal of the tumor was achieved in 2 patients and subtotal resection was received in one patient. No severe postoperative complications and sequelae occurred. In 6 months to 2 years' follow-up, the MRI showed that 2 patients had no residue tumor, and one patient died due to recurrence of the tumor 1 year after operation. CONCLUSION: The endoscopic transoral tans-posteriorwall pharynx approach might be valuable in the treatment of the hypo-clivus chordoma. The use of the endoscope allows for direct access to the hypo-clivus lesions while minimizing the chances of surrounding anatomic structure injury. In addition, this approach has the advantages of quick recovery, avoidance of catastrophic complications and sequelae. Especially, various angle view of the endoscope provides a panoramic view of the hypo-clivus, thus exposing and resecting hide lesion which can not be exposed by other approaches. This approach might facilitate complete resection of the chordoma with maximal preservation of normal tissues. PMID- 17923051 TI - Chronic lateral ankle instability and associated conditions: a rationale for treatment. AB - BACKGROUND: Ankle sprains have a high incidence of associated injuries and conditions that may be unrecognized at the initial time of injury. Failure to treat these conditions at the index surgery may compromise outcomes and delay recovery. The purpose of this study was to determine the type and frequency of associated injuries and conditions in military patients with chronic lateral ankle instability. METHODS: Between 1996 and 2002, 160 patients had 180 modified Brostrom-Gould lateral ankle ligament reconstructions for chronic ankle instability. A retrospective review of the clinical history, physical examination, radiographs, and intraoperative findings was conducted. RESULTS: The overall incidence of associated extra-articular conditions and injuries found in this study was 64%; 115 conditions were identified in 180 ankles. Peroneal tendon injuries occurred with the highest frequency (28%), followed by os trigonum lesions (13%), lateral gutter ossicles (10%), hindfoot varus alignment (8%), anterior tibial spurs (3%), and tarsal coalitions (2%). Twenty revision lateral ankle ligament reconstructions were required for either persistent pain or recurrent instability. The most common associated conditions were undiagnosed hindfoot varus alignment abnormalities (28%) followed by untreated peroneal injuries (25%). CONCLUSIONS: This study confirms the frequency of conditions associated with lateral ankle instability and emphasizes several conditions that have received little attention in the literature. Identifying these associated conditions before surgery enables the surgeon to treat all conditions at one operation, returning the patient to full activity sooner. Guidelines are presented to assist clinicians in screening patients for these associated conditions. PMID- 17923052 TI - Anterior tibial tendon rupture: results of operative treatment. AB - BACKGROUND: Anterior tibial tendon ruptures are rare, and most studies have reported subjective outcome data, with little or no objective analysis. The purpose of this study was to review the results of the operative treatment of anterior tibial tendon ruptures using the American Orthopaedic Foot and Ankle Society (AOFAS) Ankle-Hindfoot Scale and objective isokinetic testing. METHODS: We retrospectively reviewed the results of operatively treated anterior tibial tendon ruptures in 10 patients. The average age at the time of surgery was 57 (18 to 79) years. The etiology of rupture was traumatic in five and spontaneous in five patients. Evaluation consisted of preoperative and postoperative questionnaires, physical examination, and isokinetic strength testing. Average time between surgery and isokinetic testing was 27.9 months. RESULTS: The average AOFAS score was 71.9 preoperatively and 89.8 postoperatively. Eight of 10 patients reported improvement in pain, and nine of 10 patients reported increased activity level postoperatively. All patients were satisfied with the overall function of their foot and would undergo the procedure again. The peak torque generated in the operative extremity during ankle dorsiflexion and hindfoot inversion was less than that of the uninvolved extremity. No statistically significant difference was noted between peak torque generation in ankles treated with direct anterior tibial tendon repair and ankles treated with anterior tibial tendon repair with augmentation. CONCLUSIONS: Operative treatment of anterior tibial tendon rupture resulted in a high level of patient satisfaction; however, isokinetic testing demonstrated a decrease in dorsiflexion and inversion strength compared to the uninjured extremity. The clinical significance of this residual weakness was not apparent in most patients. Patients with anterior tibial tendon ruptures should be forewarned that normal strength may not be a realistic expectation after surgery. PMID- 17923053 TI - Off-loading of hindfoot and midfoot neuropathic ulcers using a fiberglass cast with a metal stirrup. AB - BACKGROUND: This study was designed to assess the effectiveness of a method of off-loading large neuropathic ulcers of the hindfoot and midfoot. The device used is composed of a fiberglass cast with a metal stirrup and a window around the ulcer. METHODS: A retrospective study of 14 diabetic and nondiabetic patients was performed. All had chronic plantar hindfoot or midfoot neuropathic ulcers that failed to heal with conventional treatment methods. A fiberglass total contact cast with a metal stirrup was applied. A window was made over the ulcer to allow daily ulcer care. RESULTS: The average duration of ulcer before application of the metal stirrup was 26 + 13.2 (range 7 to 52) months. The ulcer completely healed in 12 of the 14 patients treated. The mean time for healing was 10.8 weeks for midfoot ulcers and 12.3 weeks for heel ulcers. Complications developed in four patients: three developed superficial wounds and one developed a full thickness wound. In three of these four patients, local wound care was initiated, and the stirrup cast was continued to complete healing of the primary ulcer. CONCLUSIONS: A fiberglass cast with a metal stirrup is an effective off-loading device for midfoot and hindfoot ulcers. It is not removable and does not depend on patient compliance. The window around the ulcer allows for daily wound care, drainage of the ulcer and the use of vacuum-assisted closure (VAC) treatment. The complication rate is comparable to that of total contact casting. PMID- 17923054 TI - Immediate effects of silicone insoles on gait pattern in patients with flexible flatfoot. AB - BACKGROUND: Flatfoot in which a normal arch fails to develop is a common deformity in both children and adults. A frequently-used treatment is an over-the counter insole to normalize foot mechanics and relieve pain. This study was designed to evaluate the effects of over-the-counter silicone insoles on the gait patterns of patients with flexible flatfoot. METHODS: Thirty-four adults (24 women and nine men, average age 43.7 +/- 9.7 years) with bilateral symptomatic flatfoot deformities were included in the study. Flatfoot was diagnosed by a lateral talometatarsal angle of more than 4 degrees and a talocalcaneal angle of more than 30 degrees. Three-dimensional gait analysis and video recordings were done at a single session. All patients walked at self-selected speeds over a 10 meter walkway with and without insoles. Time-distance parameters and kinematic and kinetic characteristics of gait in the sagittal plane were evaluated by a quantitative gait analysis system. RESULTS: Mean lateral talometatarsal and talocalcaneal angles were 6.3 +/- 2.5 degrees and 56.1 +/- 8.6 degrees, respectively. There was no difference in gait parameters with or without the insoles. CONCLUSIONS: Over-the-counter insoles have no beneficial effect in normalizing forces acting on the foot and on the entire lower extremity in adults with flexible flatfoot. PMID- 17923055 TI - The physical characteristics of materials used in the manufacture of orthoses for patients with diabetes. AB - BACKGROUND: Neuropathic diabetic foot ulceration may be prevented if the mechanical stress transmitted to the plantar tissues can be modified. Orthotic therapy is one practical method commonly used to maintain tissue integrity. Orthotic design must consider the materials chosen for use in fabrication and profile of the device because both aspects influence the performance and durability of the device. Published research evaluating the physical properties of materials commonly used in the manufacture of orthoses for patients with diabetes is limited. This study investigated the physical properties of materials used to fabricate orthoses designed for the prevention of neuropathic diabetic foot ulcers. METHODS: Fifteen commonly used orthotic materials were selected for testing: four specifications of 6.4-mm Poron (Rogers Corp., Gent, Belgium), 3.2 mm Poron, three densities of 12-mm Ethylene Vinyl Acetate (EVA), 12-mm low density plastazote, two depths (6.4-mm, 3.2-mm) of Cleron (Algeo Ltd., Liverpool, UK), Professional Protective Technology (PPT), and MaxaCane (Algeo Ltd, Liverpool, UK). The density, resilience, stiffness, static coefficient of friction, durability, and compression set of each material were tested, ranked, and allocated a performance indicator score. RESULTS: The most clinically desirable dampening materials tested were Poron 96 (6-mm) and Poron 4000 (6-mm). High density EVA (Algeo Ltd., Liverpool, UK) and Lunacell Nora EVA (Freudenberg, Weinhein, Germany) possessed the properties most suitable to achieve motion control. The data present a simple and useful comparison and classification of the selected materials. CONCLUSIONS: Although this information should not be used as a single indicator for assessing the suitability of an orthotic material, the results provide clinically relevant information relating to the physical properties of orthotic materials commonly used in the prevention of neuropathic diabetic foot ulcers. PMID- 17923056 TI - Three-year followup study of topical glyceryl trinitrate treatment of chronic noninsertional Achilles tendinopathy. AB - BACKGROUND: Topical glyceryl trinitrate treatment has demonstrated short-term efficacy in chronic noninsertional Achilles tendinopathy. No long-term followup is reported. We aimed to assess if the demonstrated efficacy of this treatment persisted 3 years after discontinuation of therapy. METHODS: A follow-up study of 52 patients (68 tendons) treated with 6 months of glyceryl trinitrate therapy or placebo was performed 3 years after cessation of therapy. Assessment included pain scores, return to previous activity, the Victorian Institute of Sport Achilles tendon scale (VISA-A), asymptomatic patient outcomes, clinical assessment of tendon tenderness, and functional hop test. RESULTS: Patients treated with topical glyceryl trinitrate had significantly less Achilles tendon tenderness (p = 0.03), and improved VISA-A scores (p = 0.04) than those in the placebo group; 88% (28 of 32 tendons) of patients were completely asymptomatic at 3 years (VISA-A score of 100) compared to 67% (24 of 36 tendons) of patients treated with rehabilitation alone (p = 0.03 with Chi square analysis). Other outcome measures showed nonsignificant trends towards improvement in the glyceryl trinitrate group (pain scores p = 0.07, functional hop test p = 0.07, and return to sport p = 0.16). The mean estimated effect size for all outcome measures was 0.21. CONCLUSIONS: Topical glyceryl trinitrate treatment has demonstrated efficacy in treating chronic noninsertional Achilles tendinopathy, and the treatment benefits continue at 3 years. Significant differences in asymptomatic patient outcomes for the glyceryl trinitrate group continue at 3 years, and this is confirmed by the effect size estimate. This suggests that the mechanism of action of topical glyceryl trinitrate on chronic tendinopathies is more than an analgesic effect. PMID- 17923057 TI - Current trends in thromboprophylaxis in surgery of the foot and ankle. AB - BACKGROUND: Few studies exist to guide the best practice in thromboprophylaxis after foot and ankle surgery. A survey of foot and ankle surgeons was performed to assess current trends in thromboprophylaxis. METHODS: An email-based survey of American and British foot and ankle surgeons was conducted. Surgeons were questioned as to their use and type(s) of thromboprophylaxis as well as reasons for not using prophylaxis. Surgeons also were asked about their use of thromboprophylaxis in hip and knee arthroplasty, if they did these surgeries. RESULTS: Nearly one-fifth (19%, 27) of surgeons routinely used thromboprophylaxis in both elective and trauma foot and ankle surgery. The most common situation for use was in a postoperative patient who was immobilized and nonweightbearing. A lack of published evidence and a low rate of thromboembolism were the most commonly cited reasons for not using thromboprophylaxis. CONCLUSIONS: This survey showed a wide variability in thromboembolic prophylaxis. It suggests that despite the literature indicating to the contrary, a significant proportion of foot and ankle surgeons routinely use thromboprophylaxis. Confusion remains regarding the appropriateness of thromboprophylaxis and what type(s)(if any) should be used. This study has identified a need for more in-depth evaluation of the importance of, and possible prophylaxis against, thromboembolic problems after foot and ankle surgery. PMID- 17923058 TI - Biomechanical testing of epitenon suture strength in Achilles tendon repairs. AB - BACKGROUND: Recent evidence that early, active mobilization protocols after Achilles tendon repairs increase recovery speed and strength make operative repair strength critical to positive outcomes after Achilles tendon ruptures. While previous research has focused on core (tendon proper) repair techniques, no previous literature has reported testing of core repairs augmented with epitenon sutures, which have been shown to increase the strength of repairs of flexor tendons of the hand. METHODS: Five matched pairs of fresh frozen human Achilles tendons were tested with and without the addition of an epitenon suture to the core repair suture. All specimens were repaired using a No. 2 Ethibond Krakow locking loop core suture. The epitenon suture was added to one tendon randomly chosen from each pair, using a 4-0 nylon suture. All specimens were mounted on an MTS testing machine (MTS Systems Corp., Eden Prairie, MN) and loaded to failure, which was defined as a 1-cm gap formation. RESULTS: The addition of epitenon sutures significantly increased the force necessary to produce a 2-mm gap as compared to core sutures alone by 74%, and it increased the average load to failure by 119%. Also, initial tendon stiffness was 173% greater in tendons reinforced with epitenon sutures. CONCLUSIONS: This study demonstrates that greater resistance to gap formation, approximation of tissue ends, and tensile strength were achieved by the addition of an epitenon suture. Clinical relevance may improve healing by decreased gap formation at the repair site and a lower risk of adhesion formation. PMID- 17923059 TI - Effect of pilot-hole size on the pullout strength of flexor digitorum longus transfer fixed with a bioabsorbable screw. AB - BACKGROUND: Fixation of tendon transfers with a bioabsorbable interference-fit screw has several advantages over other fixation methods: decreased dissection, operative time, and blood loss; preservation of tendon length; no interference with radiographic studies; no need for implant removal; and no barrier to revision surgery. Whether strength of fixation is affected by the size of the pilot hole has not been established. The purpose of this study was to determine the effect of pilot hole size on the pullout strength of a flexor digitorum longus (FDL) tendon secured into a bone analog using a 5.5-mm bioabsorbable screw. METHODS: Thirty FDL tendons were harvested from 15 cadaver specimens and secured into predrilled 4 x 4 x 4 cm bone cubes with a 5.5-mm Arthrex bioabsorbable screw (Arthrex, Naples, FL). The use of bone analog foam cubes ensured consistent porosity at the insertion site, eliminating the variations associated with varying bone densities of cadaver specimens. Pilot hole sizes studied were 5.0 mm, 5.5 mm, and 6.0 mm. Pullout tests were done with an servohydraulic testing frame (MTS, Eden Prairie, MN). RESULTS: There was no significant difference (p = 0.4) between the pullout forces and stresses among the three pilot hole sizes. All specimens failed at the interface between the FDL and the bioabsorbable screw. In the 6.0-mm pilot hole group, there was a trend for increased pullout strength with increased tendon size. CONCLUSIONS: With a bioabsorbable 5.5-mm screw used for FDL transfer, a pilot hole the same size or a half millimeter larger or smaller than the screw had no statistically significant effect on the strength of the construct, even with tendons of different sizes. PMID- 17923060 TI - The influence of second toe and metatarsal length on stress fractures at the base of the second metatarsal in classical dancers. AB - BACKGROUND: Stress fractures at the base of the second metatarsal frequently occur in female classical dancers. There is a strong belief that a foot shape in which the first metatarsal or toe is shorter than the second metatarsal or toe increases the risk of this injury in dancers. However, there is a lack of empirical evidence to support this theory. The objective of this study was to examine the influence of the relative length difference between the first and second metatarsals and first and second toes on the frequency of stress fractures at the base of the second metatarsal in elite, female classical dancers. METHODS: Both feet of 50 elite female classical dancers were measured for length differences between the first and second toes and first and second metatarsals. Retrospective analysis of dancers' medical histories revealed 17 feet with stress injury and 83 without. The mean of the difference between the metatarsal and toe length for the stress-injury group was compared to that of the control group. RESULTS: No difference between the groups was identified for first and second toe length difference (p = 0.865) and the relative difference between the ends of the first two metatarsals (p = 0.815). CONCLUSIONS: Dancers who had a stress injury at the base of the second metatarsal displayed similar variances in the two independent variables as dancers who had not had such an injury. PMID- 17923061 TI - Adams-Oliver syndrome: congenital disease with gait disorder: a case report. PMID- 17923062 TI - Chondrosarcoma of the calcaneus: amputation or resection with limb preservation: a case report. PMID- 17923063 TI - Mesenchymal chondrosarcoma of the talus: a case report. PMID- 17923064 TI - Semitendinosus tendon augmentation for a large defect after Achilles tendon rupture: two case reports. PMID- 17923065 TI - Clinical tip: semi-prone position for Achilles tendon surgery. PMID- 17923066 TI - Technical tip: lateral popliteal sciatic nerve block with continuous infusion for foot and ankle surgery. PMID- 17923067 TI - Surgical strategies: the Lapidus procedure. PMID- 17923068 TI - Food allergy: oral tolerance or immunotherapy? PMID- 17923069 TI - Correlation of clinical score to pulmonary function and oxygen saturation in children with asthma attack. AB - BACKGROUND: The aim of this study is to demonstrate the importance of the relation between clinical score, pulse oximetry and spirometric tests in an asthma attack. METHODS: In this randomized, double blind, observational study, 110 children (age 2-15 years) with an asthma attack who were admitted to emergency room were evaluated. Patient history, physical examination, clinical score and oxygen saturation were recorded in all patients; however pulmonary function tests were obtained only in 54 children who were over 5 years of age. The clinical score was derived from respiratory rate, wheezing, dyspnea and retractions. RESULTS: Both oxygen saturation and spirometric tests were found to be significantly correlated with the clinical score in children. CONCLUSION: The clinical score could be used for assessing the severity of the asthma attack particularly in developing countries where laboratory facilities are not available or pulmonary function tests are not feasible. PMID- 17923070 TI - Oral rush desensitization to cow milk. Following of desensitized patients during three years. AB - We report the induction of tolerance in four patients with severe IgE-mediated cow's milk allergy, with an oral rush desensitization by introducing increasing daily doses of cow's milk (CM) for 5 days under clinical conditions in order to enable the patients to tolerate 200 ml of CM daily. Our results indicate that we can induce clinical tolerance in CM allergy by oral administration of progressive doses of milk. After three years of following, the four patients are taking CM with good tolerance. Specific IgE levels of casein have decreased progressively during these three years until being not detectable in three of the four patients and also a reduction has been observed in the cutaneous skin prick test reactions to CM. PMID- 17923071 TI - Demographic, laboratory and clinical characterisation of adult portuguese asthmatic patients. AB - BACKGROUND: Asthma is a heterogeneous chronic inflammatory condition characterised by reversible airway obstruction and hyperresponsiveness associated with underlying bronchial inflammation and structural changes. It represents an increasing health problem and is a huge burden on the patients, their families and society. The aim of the study was to characterise the adult asthmatic population attending a Hospital Allergy Clinic between the years of 2003 and 2006. METHODS: Clinical files from the Allergy Outpatient Clinic of Cova da Beira Hospital were sequentially studied. The total population analysed included 335 female and 130 male asthmatic patients. Bronchial asthma was characterised by clinical history, skin prick testing to aeroallergens, determination of total and specific IgE and lung function testing, and classified according to international guidelines. RESULTS: Of the patients studied, 70 % had allergic asthma, and 30 % had non-allergic asthma. When compared to allergic asthma, non-allergic asthma was more frequently associated with older age, perennial symptoms and female gender. More allergic than non-allergic asthma patients also had rhinitis and the reverse was true regarding drug allergy and oesophageal reflux. Grass pollen and mites were the major sensitisers for allergic asthmatics. The sensitisation profile was significantly different between urban- and rural-based asthmatic patients regarding tree pollen, fungi and moulds. CONCLUSIONS: In this population, rhinitis was more frequently associated with allergic than with non allergic asthma. The two types of asthma did not differ in clinical severity or changes in lung function. Sensitisation profiles were different between the urban and rural patients. PMID- 17923072 TI - Update on the treatment of primary immunodeficiencies. AB - A general review of advances in the treatment of Primary Immunodeficiencies (PID) has been performed. Treatment with immunoglobulins is indicated in cases of humoral immunodeficiencies and in selected cases of combined immunodeficiencies. The use of intramuscular immunoglobulins in the treatment of PID was abandoned after obtaining the intravenous immunoglobulins, since these are much more effective and have fewer adverse effects. Now subcutaneous immunoglobulins are also available. Immunoglobulins help to keep the patients free of symptoms and infections as these substances are able to neutralise infectious agents, modulate and promote the immune response and favour phagocytosis. Adverse effects have been reported in 5-15 % of patients receiving IVIg, and patients with deficiencies of subclasses of IgG with IgA deficiency and/or anti-IgA antibodies are at risk of severe reactions. No severe adverse effects of subcutaneous immuneglobulins have been reported and the medication can be self-administered. The efficacy and safety of IVIg and SCIg are similar and SCIg administered at home is associated with better quality of life. Stem Cell Transplantation (SCT) in Primary Immunodeficiencies is aimed at restoring the number and/or function of lymphocytes or phagocytes. Matched, related or unrelated donors, or related haploidentical donors are selected. HLA class II mismatched unrelated donors are avoided owing to the risk of severe graft versus host disease (GVHD). Stem cells are obtained from bone marrow, cord blood or peripheral blood. Prophylactic immunossupression (as well as donor T lymphocyte depletion in haploidentical and unrelated donors) is performed to avoid or minimize GVHD. Less toxic "reduced intensity" protocols now exist for pre-transplantation conditioning, indicated to avoid graft rejection if there is residual T-lymphocyte immunity in the host. In the majority of Severe Combined Immunodeficiencies (SCID), SCT results in T lymphocytes graft and the antibody immunodeficiency persists in many cases. The results are better the earlier it is performed, with the absence of previous infections, and with the degree of matching. The patient must be maintained in a laminar flow room with broad anti-infectious prophylaxis and with the intravenous administration of gammaglobulin for a variable period. Many other complications can be expected. Gene therapy. Patients with PID are ideal candidates, as they are monogenic, the haematopoietic cells are easily obtained and virus replication is easy within them. Vectors (viruses) "infect" the stem cells of the patient's bone marrow, producing the transfection of the wild (healthy) gene in these cells. Encouraging results have been achieved in X-linked SCID as there are a number of patients who are considered "cured", although neoplastic processes have occurred due to the activation of proto-oncogenes close to the point of insertion of the external gene, using retroviruses as vectors; there are now trials with adenovirus, physical methods (direct injection...) and chemical methods (viral modification, artificial viruses...). Gene therapy has also been performed in patients with Chronic Granulomatous Disease and trials will improve in the future with changes in protocols used in oncology and infectious diseases. PMID- 17923073 TI - Risk factors associated to the prevalence of asthma in adolescence. AB - The ISAAC project (International Study of Asthma and Allergies in Childhood) was developed with the purpose of determining the prevalence of asthma in schoool children, with the definition of two age groups: infancy, when asthma is more frequent, and adolescence, when mortality associated to asthma is greater. However, the study does not address the risk factors associated with this pathology. Our aim is to comparatively analyze the prevalence of respiratory symptoms and the asthma epidemiological factors in the two age groups and in both sexes. Since the results of ISAAC Phase I are available, we can study the variations in risk factors, relating them to the current prevalence of asthma. The prevalence of bronchial asthma in our adolescents is 10 % - this implying a significant increase with respect to the data corresponding to ISAAC Phase I. Likewise, the prevalence of wheezing in the last 12 months has also increased significantly with respect to the data corresponding to ISAAC Phase I. It can be affirmed that familial antecedents of asthma are significantly correlated to the fact of having experienced asthma at some point in time, or to wheezing in the previous 12 months. In the same way as for bronchial asthma, the increase in rhinitis has been significant. The presence of animals in the home is significantly associated to the fact of having experienced asthma at some point in time, though the presence of animals in the home has decreased significantly. An encouraging finding in our study is the fact that smoking has decreased significantly among adolescents in comparison with the previous data. However, the same does not apply to smoking among parents, which shows results similar to those recorded in the previous study - with an influence upon the same habit in the offspring. As in other studies, active smoker status is seen to be associated with the fact of having experienced asthma at some point in time. Although a common observation, the importance of hyper-responsiveness with exercise among adolescents is still not acknowledged, despite its significant correlation to the fact of having experienced asthma at some point in time, or wheezing in the previous 12 months. PMID- 17923074 TI - Corticosteroids (inhaled and/or intranasal) in the treatment of respiratory allergy in children: safety vs. efficacy. AB - BACKGROUND: Topical administration of Corticosteroids (CS) can reduce the total dose of CS required to treat the patient and minimize side effects. Topical CS is extremely effective and has an excellent safety profile. Nonetheless, care must be taken when multiple sites such as lungs, nose and skin are being treated. CS mechanisms of action on the inflammatory process are complex. The aim of this study is to review such mechanisms and the adverse events secondary to it. METHODS: Review English database (Embase, PubMed, Scielo) searching words: CS, adverse events, inhaled CS, intranasal CS, and children. RESULTS: There is a classic mechanism involving a genomic effect of CS and a non-genomic effect, independently of gene transcription process. This mechanism acts by reducing mucosal blood flow in the asthmatic airways. Second-generation topical CS is the treatment of choice in allergic diseases control because of their good anti inflammatory activity, poor absorption and first-pass hepatic metabolism. When comparing different CS, it is important to compare therapeutically equivalent doses. Although topical CS reduces systemic side effects, local and even systemic side effects can occur. Many factors affect the amount of drug that reaches the lung, including inhaler technique and inhaler type, fine particle dose and particle distribution. CONCLUSION: Most patients with allergic diseases respond to CS treatment, but there is a small subset of them whose response is unsatisfactory even with high doses of CS. They are classified as corticosteroid resistant asthmatics. Pro-inflammatory cytokines appear to up regulate the expression of GRb that has been associated with CS resistance. PMID- 17923075 TI - Recommendations for administering the triple viral vaccine and antiinfluenza vaccine in patients with egg allergy. AB - Actually, food allergy is an emerging pathology; and egg allergy is the most frequent in childhood. The recommendations for measles, mumps and rubella (MMR) and influenza vaccination are increasing each year. This implementation increases the exposure of patients with egg allergy to such vaccines. In Spain, since 2004 the only available vaccine for MMR is grown in cultures of fibroblast from chick embryos; previously, patients with egg allergy were vaccinated with an alternative vaccine cultivated in diploid human cells which is no longer commercialized. Influenza vaccines grow in chick egg and the final product contains egg proteins (large variation in egg protein content has been reported). As controversy exist, the Food Allergy Committee of Spanish Society of Clinical Immunology and Pediatric Allergy decided to report some recommendations for the safe administration of MMR and influenza vaccines in patients with egg allergy. In summary, MMR vaccine is safe for children with egg allergy, only in patients with severe anaphylactic reaction after egg ingestion is recommended the administration in his reference hospital. Influenza vaccine is contraindicated in patients with severe anaphylactic reaction after egg ingestion. The rest can receive influenza vaccine in a 2-dose protocol with a vaccine that contains no more than 1.2 mcg of egg protein for mL. PMID- 17923076 TI - Immediate hypersensitivity and delayed hypersensitivity to clopidogrel. AB - Clopidogrel is a new antiplatelet prescribed for the secondary prevention of atherosclerotic events. The literature shows that the clinical manifestations of clopidogrel hypersensitivity include urticaria1, skin rash2-4, and angioedema5. The precise immunological mechanism underlying clopidogrel hypersensitivity has not been established. We describe two cases of hypersensitivity reaction due to clopidogrel. The first constituted an immediate reaction after clopidogrel intake. In this case we demonstrated type 1 hypersensitivity using cutaneous tests. The second case represented a delayed hypersensitivity reaction confirmed by oral challenge testing, in which good tolerance to other antiplatelet drugs such as ticlopidine was demonstrated. PMID- 17923077 TI - Anisakis simplex allergy and nephrotic syndrome. AB - BACKGROUND: Anisakis simplex is a helmintic parasite of fish and shellfish that can induce in humans, after consumption, an immunoallergic response with multiorganic affectation, especially cutaneous (urticaria and angioedema), gastrointestinal and anaphylactic symptoms. Nephrotic syndrome can be produced by helmintic infection, especially schistosomiasis. OBJECTIVE: To report the unusual case of nephrotic syndrome associated to an allergic response to A. simplex. METHODS: A 60-year-old woman suffered from nephrotic syndrome three weeks after a generalized urticaria, angioedema and nauseas episode, following a raw anchovy's ingestion. Etiological study of nephrotic syndrome and urticaria/angioedema, cutaneous test, serum total and specific IgE determination, IgE-immunoblotting, and HLA typing were performed for diagnostic evaluation. RESULTS: Prick-test with A. simplex extract was positive. It was observed a significance increase of total IgE to the three weeks of anchovy's ingestion and then it diminished progressively. Specific IgE to A. simplex was > 100 UI/ml. Immunoblotting to A. simplex, revealed the existence of IgE-binding proteins with molecular mass ranging from 20 kDa to 99 kDa. Class I HLA expressed by the patient was type HLA B12. Other secondary causes of nephrotic syndrome were carefully ruled out. CONCLUSIONS: These data suggest, for the first time, the association between allergy to A. simplex and nephrotic syndrome. PMID- 17923078 TI - Seeking an international dialogue on research integrity. AB - Scientific misconduct is a global problem, yet protocols for addressing it remain highly fragmented and uneven. A conference held last month in Lisbon aimed to encourage international efforts to promote research integrity and to prevent misconduct. PMID- 17923079 TI - Retrovirus silencing by an epigenetic TRIM. AB - Embryonic cells silence transcription by retroviruses, but how do they recognize virus DNA? In this issue, Wolf and Goff (2007) report that a TRIM28 corepressor complex binds to the retrovirus primer binding site. Epigenetic silencing of retrovirus transcription is accomplished by "writing" a dimethyl mark on lysine 9 of histone H3 that is read by the heterochromatin protein HP1gamma. PMID- 17923080 TI - Loss of T-bet sends host-microbe mutualism awry. AB - Immune homeostasis in the intestine involves complex interactions between microbial organisms and host cells. In this issue, Garrett et al. (2007) describe a communicable form of colitis that is induced by deficiency of the transcription factor T-bet in cells of the innate immune system. PMID- 17923081 TI - How to assemble a capsid. AB - Retroviral capsids are composed of hexagonal arrays of the viral CA protein. In this issue of Cell, Ganser-Pornillos et al. (2007) provide a molecular model of the hexagonal HIV-1 CA lattice obtained from a new electron cryomicroscopic reconstruction. This study reveals the three principal stabilizing interfaces in the capsid lattice and explains how two different classes of inhibitors can block capsid assembly. PMID- 17923082 TI - Dynamic regulation of the INAD signaling scaffold becomes crystal clear. AB - PDZ domains are common building blocks of scaffold proteins that enhance specificity and speed in signal transduction cascades. Although PDZ modules are often viewed as passive participants, Mishra et al. (2007) now show that a PDZ domain in INAD, a scaffold protein in photoreceptor cells of the fruit fly, undergoes a light-dependent conformational change, which has important consequences for signaling and animal behavior. PMID- 17923083 TI - miRNAs play a tune. AB - Two new studies describe functionally relevant interactions between microRNAs (miRNAs) and their targets in the immune system and the brain (Xiao et al., 2007; Karres et al., 2007). Furthermore, these studies illustrate the involvement of miRNAs in tuning the expression of target genes to physiologically relevant levels. PMID- 17923084 TI - miRNAs get an early start on translational silencing. AB - MicroRNAs (miRNAs) control gene expression by regulating mRNA stability and translation. Using cell-free in vitro systems, several labs have recently reported insights into the molecular mechanisms underlying miRNA-guided translational repression (Kiriakidou et al., 2007; Mathonnet et al., 2007; Thermann and Hentze, 2007; Wakiyama et al., 2007). These new findings indicate that miRNAs inhibit translation at early steps of initiation. PMID- 17923085 TI - H3K27 demethylases, at long last. AB - Methylation of lysine 27 on histone H3 (H3K27me) by the Polycomb complex (PRC2) proteins is associated with gene silencing in many developmental processes. A cluster of recent papers (Agger et al., 2007; De Santa et al., 2007; Lan et al., 2007; Lee et al., 2007) identify the JmjC-domain proteins UTX and JMJD3 as H3K27 specific demethylases that remove this methyl mark, enabling the activation of genes involved in animal body patterning and the inflammatory response. PMID- 17923086 TI - Communicable ulcerative colitis induced by T-bet deficiency in the innate immune system. AB - Inflammatory bowel disease (IBD) has been attributed to overexuberant host immunity or the emergence of harmful intestinal flora. The transcription factor T bet orchestrates inflammatory genetic programs in both adaptive and innate immunity. We describe a profound and unexpected function for T-bet in influencing the behavior of host inflammatory activity and commensal bacteria. T-bet deficiency in the innate immune system results in spontaneous and communicable ulcerative colitis in the absence of adaptive immunity and increased susceptibility to colitis in immunologically intact hosts. T-bet controls the response of the mucosal immune system to commensal bacteria by regulating TNF alpha production in colonic dendritic cells, critical for colonic epithelial barrier maintenance. Loss of T-bet influences bacterial populations to become colitogenic, and this colitis is communicable to genetically intact hosts. These findings reveal a novel function for T-bet as a peacekeeper of host-commensal relationships and provide new perspectives on the pathophysiology of IBD. PMID- 17923087 TI - TRIM28 mediates primer binding site-targeted silencing of murine leukemia virus in embryonic cells. AB - Moloney murine leukemia virus (M-MLV) replication is restricted in embryonic carcinoma (EC) and embryonic stem (ES) cells, likely to protect the germ line from insertional mutagenesis. Proviral DNAs are potently silenced at the level of transcription in these cells. This silencing is largely due to an unidentified trans-acting factor that is thought to bind to the primer binding site (PBS) of M MLV and repress transcription from the viral promoter. We have partially purified a large PBS-mediated silencing complex and identified TRIM28 (Kap-1), a known transcriptional silencer, as an integral component of the complex. We show that RNAi-mediated knockdown of TRIM28 in EC and ES cells relieves the restriction and that TRIM28 is bound to the PBS in vivo when restriction takes place. The identification of TRIM28 as a retroviral silencer adds to the growing body of evidence that many TRIM family proteins are involved in retroviral restriction. PMID- 17923088 TI - Structure of full-length HIV-1 CA: a model for the mature capsid lattice. AB - The capsids of mature retroviruses perform the essential function of organizing the viral genome for efficient replication. These capsids are modeled as fullerene structures composed of closed hexameric arrays of the viral CA protein, but a high-resolution structure of the lattice has remained elusive. A three dimensional map of two-dimensional crystals of the R18L mutant of HIV-1 CA was derived by electron cryocrystallography. The docking of high-resolution domain structures into the map yielded the first unambiguous model for full-length HIV-1 CA. Three important protein-protein assembly interfaces are required for capsid formation. Each CA hexamer is composed of an inner ring of six N-terminal domains and an outer ring of C-terminal domains that form dimeric linkers connecting neighboring hexamers. Interactions between the two domains of CA further stabilize the hexamer and provide a structural explanation for the mechanism of action of known HIV-1 assembly inhibitors. PMID- 17923089 TI - Dynamic scaffolding in a G protein-coupled signaling system. AB - The INAD scaffold organizes a multiprotein complex that is essential for proper visual signaling in Drosophila photoreceptor cells. Here we show that one of the INAD PDZ domains (PDZ5) exists in a redox-dependent equilibrium between two conformations--a reduced form that is similar to the structure of other PDZ domains, and an oxidized form in which the ligand-binding site is distorted through formation of a strong intramolecular disulfide bond. We demonstrate transient light-dependent formation of this disulfide bond in vivo and find that transgenic flies expressing a mutant INAD in which PDZ5 is locked in the reduced state display severe defects in termination of visual responses and visually mediated reflex behavior. These studies demonstrate a conformational switch mechanism for PDZ domain function and suggest that INAD behaves more like a dynamic machine rather than a passive scaffold, regulating signal transduction at the millisecond timescale through cycles of conformational change. PMID- 17923090 TI - Acetylation-dependent signal transduction for type I interferon receptor. AB - Cytokine-activated receptors initiate intracellular signaling by recruiting protein kinases that phosphorylate the receptors on tyrosine residues, thus enabling docking of SH2 domain-bearing activating factors. Here we report that in response to type 1 interferon (IFNalpha), IFNalpha receptors recruit cytoplasmic CREB-binding protein (CBP). By binding to IFNalphaR2 within the region where two adjacent proline boxes bear phospho-Ser364 and phospho-Ser384, CBP acetylates IFNalphaR2 on Lys399, which in turn serves as the docking site for interferon regulatory factor 9 (IRF9). IRF9 interacts with the acetyl-Lys399 motif by means of its IRF homology2 (IH2) domain, leading to formation of the ISGF3 complex that includes IRF9, STAT1, and STAT2. All three components are acetylated by CBP. Remarkably, acetylation within the DNA-binding domain (DBD) of both IRF9 and STAT2 is critical for the ISGF3 complex activation and its associated antiviral gene regulation. These results have significant implications concerning the central role of acetylation in cytokine receptor signal transduction. PMID- 17923091 TI - Insights into Hsp70 chaperone activity from a crystal structure of the yeast Hsp110 Sse1. AB - Classic Hsp70 chaperones assist in diverse processes of protein folding and translocation, and Hsp110s had seemed by sequence to be distant relatives within an Hsp70 superfamily. The 2.4 A resolution structure of Sse1 with ATP shows that Hsp110s are indeed Hsp70 relatives, and it provides insight into allosteric coupling between sites for ATP and polypeptide-substrate binding in Hsp70s. Subdomain structures are similar in intact Sse1(ATP) and in the separate Hsp70 domains, but conformational dispositions are radically different. Interfaces between Sse1 domains are extensive, intimate, and conservative in sequence with Hsp70s. We propose that Sse1(ATP) may be an evolutionary vestige of the Hsp70(ATP) state, and an analysis of 64 mutant variants in Sse1 and three Hsp70 homologs supports this hypothesis. An atomic-level understanding of Hsp70 communication between ATP and substrate-binding domains follows. Requirements on Sse1 for yeast viability are in keeping with the distinct function of Hsp110s as nucleotide exchange factors. PMID- 17923092 TI - Diverse cellular functions of the Hsp90 molecular chaperone uncovered using systems approaches. AB - A comprehensive understanding of the cellular functions of the Hsp90 molecular chaperone has remained elusive. Although Hsp90 is essential, highly abundant under normal conditions, and further induced by environmental stress, only a limited number of Hsp90 "clients" have been identified. To define Hsp90 function, a panel of genome-wide chemical-genetic screens in Saccharomyces cerevisiae were combined with bioinformatic analyses. This approach identified several unanticipated functions of Hsp90 under normal conditions and in response to stress. Under normal growth conditions, Hsp90 plays a major role in various aspects of the secretory pathway and cellular transport; during environmental stress, Hsp90 is required for the cell cycle, meiosis, and cytokinesis. Importantly, biochemical and cell biological analyses validated several of these Hsp90-dependent functions, highlighting the potential of our integrated global approach to uncover chaperone functions in the cell. PMID- 17923093 TI - The conserved microRNA miR-8 tunes atrophin levels to prevent neurodegeneration in Drosophila. AB - microRNAs (miRNAs) bind to specific messenger RNA targets to posttranscriptionally modulate their expression. Understanding the regulatory relationships between miRNAs and targets remains a major challenge. Many miRNAs reduce expression of their targets to inconsequential levels. It has also been proposed that miRNAs might adjust target expression to an optimal level. Here we analyze the consequences of mutating the conserved miRNA miR-8 in Drosophila. We identify atrophin as a direct target of miR-8. miR-8 mutant phenotypes are attributable to elevated atrophin activity, resulting in elevated apoptosis in the brain and in behavioral defects. Reduction of atrophin levels in miR-8 expressing cells to below the level generated by miR-8 regulation is detrimental, providing evidence for a "tuning target" relationship between them. Drosophila atrophin is related to the atrophin family of mammalian transcriptional regulators, implicated in the neurodegenerative disorder DRPLA. The regulatory relationship between miR-8 and atrophin orthologs is conserved in mammals. PMID- 17923094 TI - MiR-150 controls B cell differentiation by targeting the transcription factor c Myb. AB - MiR-150 is a microRNA (miRNA) specifically expressed in mature lymphocytes, but not their progenitors. A top predicted target of miR-150 is c-Myb, a transcription factor controlling multiple steps of lymphocyte development. Combining loss- and gain-of-function gene targeting approaches for miR-150 with conditional and partial ablation of c-Myb, we show that miR-150 indeed controls c Myb expression in vivo in a dose-dependent manner over a narrow range of miRNA and c-Myb concentrations and that this dramatically affects lymphocyte development and response. Our results identify a key transcription factor as a critical target of a stage-specifically expressed miRNA in lymphocytes and suggest that this and perhaps other miRNAs have evolved to control the expression of just a few critical target proteins in particular cellular contexts. PMID- 17923095 TI - Emotion enhances learning via norepinephrine regulation of AMPA-receptor trafficking. AB - Emotion enhances our ability to form vivid memories of even trivial events. Norepinephrine (NE), a neuromodulator released during emotional arousal, plays a central role in the emotional regulation of memory. However, the underlying molecular mechanism remains elusive. Toward this aim, we have examined the role of NE in contextual memory formation and in the synaptic delivery of GluR1 containing alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA)-type glutamate receptors during long-term potentiation (LTP), a candidate synaptic mechanism for learning. We found that NE, as well as emotional stress, induces phosphorylation of GluR1 at sites critical for its synaptic delivery. Phosphorylation at these sites is necessary and sufficient to lower the threshold for GluR1 synaptic incorporation during LTP. In behavioral experiments, NE can lower the threshold for memory formation in wild-type mice but not in mice carrying mutations in the GluR1 phosphorylation sites. Our results indicate that NE-driven phosphorylation of GluR1 facilitates the synaptic delivery of GluR1 containing AMPARs, lowering the threshold for LTP, thereby providing a molecular mechanism for how emotion enhances learning and memory. PMID- 17923096 TI - Global analysis of mRNA localization reveals a prominent role in organizing cellular architecture and function. AB - Although subcellular mRNA trafficking has been demonstrated as a mechanism to control protein distribution, it is generally believed that most protein localization occurs subsequent to translation. To address this point, we developed and employed a high-resolution fluorescent in situ hybridization procedure to comprehensively evaluate mRNA localization dynamics during early Drosophila embryogenesis. Surprisingly, of the 3370 genes analyzed, 71% of those expressed encode subcellularly localized mRNAs. Dozens of new and striking localization patterns were observed, implying an equivalent variety of localization mechanisms. Tight correlations between mRNA distribution and subsequent protein localization and function, indicate major roles for mRNA localization in nucleating localized cellular machineries. A searchable web resource documenting mRNA expression and localization dynamics has been established and will serve as an invaluable tool for dissecting localization mechanisms and for predicting gene functions and interactions. PMID- 17923098 TI - Manipulating response set in a choice reaction task: evidence for anatomical coding in response selection. AB - Accurate motor performance requires a process of response selection that chooses the correct response out of a set of possible ones. Most theories of response selection assume that this selection process operates on spatial codes, which define the location of stimuli and responses in environmental coordinates, with little or no role for the anatomical codes of the effectors involved. In this study, I systematically manipulated the anatomical identities of the response fingers in a four-choice reaction task, while keeping the stimulus and response locations constant. Results showed that homologous fingers on different hands modulated the reaction time profiles and error patterns in a mirror-symmetrical way. This finding indicates that anatomical factors may play a more substantial role in response selection than often assumed. PMID- 17923097 TI - Forkhead transcription factors II. PMID- 17923099 TI - A miniaturized high-throughput screening assay for fucosyltransferase VII. AB - Fucosyltransferase VII (FucTVII) is a very promising drug target for treatment of inflammatory skin diseases. Its activity is required for synthesis of the sialyl Lewis X glycoepitopes on the E- and P-selectin ligands, necessary for lymphocyte migration into the skin. High-throughput screening (HTS) of large chemical libraries has become the main source of novel chemical entities for the pharmaceutical industry. The screening of very large compound collections requires the use of specialized assay techniques that minimize time and costs. We describe the development of a miniaturized scintillation proximity assay for human FucTVII based on a oligosaccharide acceptor substrate that is identical to the glycosylation of the physiological substrate. In addition to assay development, the assay performance in a HTS campaign is shown. We screened 798,131 compounds from the Schering AG HTS library and identified 233 IC50 hits; 229 hits were FucTVII specific in so far as they did not inhibit either alpha fucosidase or galactosyltransferase. In addition to screening a drug-like small molecule collection, we worked on rational approaches to develop inhibitors or glycosidic decoys based on oligosaccharide-substrate analogues. The structure activity relationship observed thereby is very narrow and shows strict requirements that are consistent with the described substrate specificity of FucTVII. PMID- 17923101 TI - Molecular beacons for isothermal fluorescence enhancement by the cleavage of RNase HII from Chlamydia pneumoniae. AB - This article describes a new assay for isothermal enhancement of fluorescence intensity. The assay is based on the cleavage of duplexes formed by the chimeric DNA-rN(1)-DNA molecular beacon (cMB) and target DNA with Chlamydia pneumoniae RNase HII (CpRNase HII). The loop sequence of the cMB, which was designed according to the target sequence, contains a single ribonucleotide. The combination of CpRNase HII cleavage and cMB (RHMB) permitted a 90-fold increase in fluorescence intensity change compared with the hybridization reaction in the presence of the same amount of target DNA. These results indicate that the RHMB assay can enhance the fluorescence signal in real-time monitoring of the target DNA. PMID- 17923100 TI - Apolipoprotein E isoform-specific binding to the low-density lipoprotein receptor. AB - Apolipoprotein E (apoE) is a ligand for members of the low-density lipoprotein receptor (LDLR) family and functions in plasma cholesterol homeostasis. A fluorescence-based assay has been employed in molecular studies of receptor ligand interactions. Competition experiments revealed isoform-specific differences in binding of lipid-associated apoE N terminal (NT) domain to a recombinant soluble LDLR (sLDLR). In a similar manner, lipid--associated-but not lipid-free--full-length apoE3 showed binding activity to sLDLR. The molecular chaperone, receptor-associated protein, inhibited apoE3-NT-phospholipid complex binding to sLDLR. Kinetic studies of apoE3-NT-phospholipid complex interaction with sLDLR revealed time-dependent effects of apoE-NT isoform binding to sLDLR. The results reveal a discerning method for study of the molecular basis of ligand interactions that likely influence receptor function in maintenance of whole body cholesterol homeostasis. PMID- 17923102 TI - Fluoroscopic assessment of protein leakage during Xenopus oocytes in-cell NMR experiment by co-injected EGFP. PMID- 17923103 TI - Improved methods for the preparation of [3H]folate polyglutamates: biosynthesis with Lactobacillus casei and enzymatic synthesis with Escherichia coli folylpolyglutamate synthetase. AB - Tritiated forms of polyglutamyl folates are not commercially available but are often needed for experimental uses in folate biochemistry. Thus, considerable interest exists in the preparation of polyglutamyl [(3)H]folates from the commercial monoglutamyl [(3)H]folates. However, refinement of established enzymatic and biological synthesis methods is still needed. To address this need we developed improved procedures for the conversion of monoglutamyl [(3)H]folates to various polyglutamyl forms. In the bacterial synthesis, Lactobacillus casei was grown in the presence of 1 ng/ml (2.27 nM) [(3)H]folic acid in Folic Acid Casei Medium. Washed cells were resuspended in 2% sodium ascorbate containing 10mM beta-mercaptoethanol and heated to release the folates. The extracted [(3)H]folates were purified on a folate-binding protein affinity column and then applied to a Sephadex G-10 column to separate the eluted poly- from the monoglutamyl folate species. High performance liquid chromatography with multichannel electrochemical detection indicated that the bacterial synthesis yielded predominantly polyglutamates of [(3)H]5-methyltetrahydrofolate and [(3)H]5-formyltetrahydrofolate (di- through heptaglutamates). The alternative method consisted of enzymatic polyglutamylation of [(3)H]folic acid catalyzed by recombinant Escherichia coli folylpolyglutamate synthetase. This enzymatic synthesis yielded predominantly tri-, tetra-, and pentaglutamyl species for the [(3)H]folate substrate. PMID- 17923104 TI - Vasodilatation of sulfur dioxide derivatives and signal transduction. AB - The vasodilatation of sulfur dioxide (SO(2)) derivatives on the rat thoracic aortic rings and its cell signal transduction pathway were studied. The levels of cAMP, cGMP, PGI(2), TXA(2) and activities of PKA and adenylyl cyclase (AC) in the rings exposed to SO(2) derivatives were determined. The results indicated that SO(2) derivatives could dose-dependently relax the isolated rat aorta rings with or without endothelium precontracted by NE, no difference was found between the rings with and without endothelium; Levels of cAMP, PGI(2), AC activity and PKA activity in the aortic rings were significantly increased by the derivatives in a dose-related way; No change of cGMP and TXA(2) levels in rings was observed; cAMP/cGMP and PGI(2)/TXA(2) ratios were increased significantly by the SO(2) derivatives. These results led to the conclusions that SO(2) derivatives might cause the endothelium-independent vasorelaxation effect, and the vasorelaxation was mediated in partly by the signal transduction pathway of PGI(2)-AC-cAMP-PKA. PMID- 17923105 TI - The nature of the transition state ensemble and the mechanisms of protein folding: a review. PMID- 17923106 TI - Fructose-2,6-bisphosphate counteracts guanidinium chloride-, thermal-, and ATP induced dissociation of skeletal muscle key glycolytic enzyme 6-phosphofructo-1 kinase: A structural mechanism for PFK allosteric regulation. AB - Rabbit muscle 6-phosphofructo-1-kinase (PFK) is the key glycolytic enzyme being regulated by diverse molecules and signals. This enzyme may undergo a reversible dissociation from a fully active homotetramer to a quite inactive dimer. There are evidences that some positive and negative modulators of PFK, such as ADP and citrate, may interfere with the enzyme oligomeric structure shifting the tetramer dimer equilibrium towards opposite orientations, where the negative modulators favor the dissociation of tetramers into dimers and vice versa. PFK is allosterically inhibited by ATP at its physiological range of concentration, an effect counteracted by fructose-2,6-bisphosphate (F2,6BP). However, the structural molecular mechanism by which ATP and F2,6BP regulate PFK is hitherto demonstrated. The present paper aimed at demonstrating that either the ATP induced inhibition of PFK and the reversion of this inhibition by F2,6BP occur through the same molecular mechanism, i.e., the displacement of the oligomeric equilibrium of the enzyme. This conclusion is arrived assessing the effects of ATP and F2,6BP on PFK inactivation through two distinct ways to dissociate the enzyme: (a) upon incubation at 50 degrees C, or (b) incubating the enzyme with guanidinium hydrochloride (GdmCl). Our results reveal that temperature- and GdmCl induced inactivation of PFK prove remarkably more effective in the presence 5mM ATP than in the absence of additives. On the other hand, the presence of 100 nM F2,6BP attenuate the effects of both high-temperature exposition and GdmCl on PFK, even in the simultaneous presence of 5mM ATP. These data support the hypothesis that ATP shifts the oligomeric equilibrium of PFK towards the smaller conformations, while F2,6BP acts in the opposite direction. This conclusion leads to important information about the molecular mechanism by which PFK is regulated by these modulators. PMID- 17923107 TI - Circular dichroism investigation of the effect of plasmid DNA structure on retention in histidine chromatography. AB - The effect of temperature-induced changes in secondary and tertiary structures of plasmid DNA (pDNA) and on the retention behaviour of open circular (oc) and supercoiled (sc) isoforms in histidine-agarose chromatography was investigated by Circular dichroism (CD) spectroscopy. Chromatographic experiments performed with three plasmids (2.7, 6.1 and 7.4kbp) and with a decreasing ammonium sulphate gradient (2.3--2.0M) showed that the retention of sc pDNA increased as temperature decreased from 24 to 5 degrees C. Such behaviour was attributed to the temperature-induced removal of negative superhelical turns in sc pDNA which is accompanied by a decrease in the number of dissociated base pairs responsible for interaction with the histidine ligands. CD spectroscopy showed that temperature has an important effect on plasmid secondary structure if adenine rich inserts are present in the plasmid structure. Chromatographic experiments also suggested that base composition could also be responsible for the induction of specific interactions with histidine ligands. PMID- 17923108 TI - The role of urgency in maladaptive behaviors. AB - Prior work on maladaptive behaviors has cited impulsivity as a risk factor. The concept of impulsivity, however, fails to address the potential role of negative affect in such behaviors. The UPPS Impulsive Behavior Scale addresses this weakness by dividing impulsivity into four subscales: Urgency, Sensation Seeking, (lack of) Premeditation, and (lack of) Perseverance. We predicted that urgency, defined as the tendency, specifically in the face of negative affect, to act quickly and without planning, would predict elevations on three maladaptive behaviors--excessive reassurance seeking, drinking to cope, and bulimic symptoms as measured by the Eating Disorder Inventory--in both cross-sectional and longitudinal analyses in an undergraduate sample (N=70). Participants were assessed at two time points, 3-4 weeks apart. Urgency significantly predicted all three outcome variables cross-sectionally at both Time 1 and Time 2. Time 1 urgency significantly predicted Time 2 excessive reassurance seeking. Changes in urgency from Time 1 to Time 2 predicted changes in all three outcome variables. Results indicate a clear cross-sectional relationship between urgency and certain maladaptive behaviors. Additionally, some form of longitudinal relationship may exist between these variables, although the use of residual change scores precluded distinction between true change and change due to error. PMID- 17923109 TI - POMT2 gene mutation in limb-girdle muscular dystrophy with inflammatory changes. AB - Defects in glycosylation of alpha-dystroglycan are associated with several forms of muscular dystrophies. Mutations in POMT2 gene have been identified in patients with congenital muscular dystrophy and brain involvement, either characterized by a Walker-Warburg/muscle-eye-brain phenotype, or by microcephaly, mental retardation, and cerebellar hypoplasia. We identified a POMT2 homozygous missense mutation in a girl with a mild limb-girdle muscular dystrophy (LGMD) phenotype, marked elevated serum creatine kinase levels, and absence of brain involvement. Muscle biopsy revealed myopathic and inflammatory changes and severe alpha dystroglycan reduction. In view of the remarkable mild clinical picture, we propose to designate this phenotype as LGMD2N. PMID- 17923110 TI - Determination of genes involved in the early process of molar root development initiation in rat by modified subtractive hybridization. AB - Rat molars and incisors have different root patterning mechanisms: the former form a multirooted pattern while the latter form a single-rooted analogue. But the genetic signals that control root patterning are poorly understood. In this study, to identify the special molecular signals which may lead to the molar root development, we firstly observed that at embryonic day 19 the molar and the incisor began differentially developing: the molar formed double-layered cells of the root sheath while the incisor formed a cervical loop. By using the subtractive hybridization method, we successfully constructed a subtraction cDNA library of the rat molar and incisor tissues. Differentially expressed gene clones were evaluated by dot-blot and sequencing. Sel1l, Nfic, Edar, GATA6, and some novel genes were found differentially expressed, which were strongly related the tooth root patterning. It is anticipated that further study of genes identified will provide insights into their specific roles in the tooth root patterning. PMID- 17923111 TI - Lysophosphatidic acid regulates inflammation-related genes in human endothelial cells through LPA1 and LPA3. AB - Lysophosphatidic acid (LPA) is a low-molecular-weight lysophospholipid (LPL), which regulates endothelial cells participating in inflammation processes via interactions with endothelial differentiation gene (Edg) family G protein-coupled receptors. In this study, we attempted to determine which LPA receptors mediate the inflammatory response in human endothelial cells. Introduction of siRNA against LPA1 significantly suppressed LPA-induced ICAM-1 mRNA, total protein, and cell surface expressions, and subsequent U937 monocyte adhesion to LPA-treated human umbilical endothelial cells (HUVECs). By knock down of LPA1 and LPA3 in HUVECs, LPA-enhanced IL-1beta mRNA expression was significantly attenuated. Moreover, LPA1 and LPA3 siRNA also inhibited LPA-enhanced IL-1-dependent long term IL-8 and MCP-1 mRNA expression, and subsequent THP-1 cell chemotaxis toward LPA-treated HUVEC-conditioned media. These results suggest that the expression of LPA-induced inflammatory response genes is mediated by LPA1 and LPA3. Our findings suggest the possible utilization of LPA1 or LPA3 as drug targets to treat severe inflammation. PMID- 17923112 TI - Bcl-xL blocks high dose doxorubicin-induced apoptosis but not low dose doxorubicin-induced cell death through mitotic catastrophe. AB - Bcl-xL is often overexpressed in human hepatocellular carcinoma cells, contributing to resistance to various chemotherapeutic agents. In this study, we investigated the role of Bcl-xL in two modes of cell death induced by different doses of doxorubicin, apoptosis and cell death through mitotic catastrophe. Bcl xL overexpression in various hepatoma cells effectively blocked apoptosis induced by high dose doxorubicin, inhibiting the loss of mitochondrial membrane potential, release of mitochondrial cytochrome c and caspase activation. Contrastingly, Bcl-xL overexpression did not block low dose doxorubicin-induced mitotic catastrophe and subsequent non-apoptotic cell death, without affecting abnormal cell cycle progression, formation of multiple micronuclei, loss of mitochondrial membrane potential, release of mitochondrial cytochrome c, and the clonogenicity of cells exposed to low dose doxorubicin. These findings indicate that low dose doxorubicin-induced cell death through mitotic catastrophe may provide an alternative therapeutic strategy for Bcl-xL-overexpressing hepatoma cells, which are resistant to pro-apoptotic treatments. PMID- 17923113 TI - Sterol biosynthesis and prokaryotes-to-eukaryotes evolution. AB - It has been widely accepted that there exists a correlation between prokaryotes to-eukaryotes evolution and atmospheric oxygen rise. However, it is a great challenge to elucidate the mechanisms underlying the correlation. Considering the facts that cellular communication of eukaryotes depends largely on membrane functions (e.g., endo- and exocytosis) and sterols play a key role in fulfilling these functions, we propose that the biosynthesis of sterols represents a critical step in the prokaryotes-to-eukaryotes evolution. Indeed, sterol biosynthesis is nearly ubiquitous among eukaryotes, but is generally excluded by prokaryotes. More importantly, during the biosynthesis of sterols, oxygen is absolutely required. Therefore, the missing link between prokaryotes-to eukaryotes evolution and atmospheric oxygen rise is likely to reside in, at least in part, sterol biosynthesis, i.e., high atmospheric oxygen concentration facilitates the generation of sterols and thus benefits the birth of complex organisms. PMID- 17923114 TI - VEGF reduces astrogliosis and preserves neuromuscular junctions in ALS transgenic mice. AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease resulting from motor neuron loss in the spinal cord and brain stem. In the present study, we found that systemic administration of recombinant vascular endothelial growth factor (VEGF) significantly diminished astrogliosis and increased the number of neuromuscular junctions in a Cu/Zn superoxide dismutase (SOD1) transgenic mouse model of ALS. Our results thus demonstrate a novel regulatory role of VEGF on astrocytes and are suggestive of protective effects of VEGF both in the peripheral and central nervous system in the SOD1 transgenic mouse model. These findings warrant further evaluation of the mechanism(s) of regulatory effects of VEGF on neuronal and non-neuronal cells, and the relation of these events to motor neuron degeneration and the onset and progression of ALS. PMID- 17923115 TI - Dynamic change of histone H2AX phosphorylation independent of ATM and DNA-PK in mouse skin in situ. AB - Histone H2AX undergoes phosphorylation on Ser 139 (gamma-H2AX) rapidly in response to DNA double-strand breaks induced by exogenous stimuli, such as ionizing radiation. However, the endogenous phosphorylation pattern and modifier of H2AX remain unclear. Here we show that H2AX is regulated physically at the level of phosphorylation at Ser139 during a hair cycle in the mouse skin. In anagen hair follicles, gamma-H2AX-positive cells were observed in the outer root sheath (ORS) and hair bulb in a cycling inferior region but not in a permanent superficial region. In telogen hair follicles, gamma-H2AX-positive cells were only detected around the germ cell cap. In contrast, following X-irradiation, gamma-H2AX was observed in various cell types including the ORS cells in the permanent superficial region. Furthermore, gamma-H2AX-positive cells were detected in the skin of mice lacking either ATM or DNA-PK, suggesting that these kinases are not essential for phosphorylation in vivo. PMID- 17923116 TI - Regulation of [Ca2+]i oscillations in mouse pancreatic islets by adrenergic agonists. AB - [Ca2+]i oscillations are an important cellular signal in glucose-induced insulin secretion. Isolated mouse pancreatic islets show a high degree of synchronization in their oscillatory pattern. Our comprehensive analysis revealed that 63% of the islets exhibited glucose-induced oscillations with low frequency (< or = 0.5/min) and 37% with high frequency (>0.5/min). The catecholamines adrenaline and noradrenaline as well as clonidine, all known to inhibit insulin secretion, were studied for their ability to modulate the glucose-induced slow large amplitude [Ca2+]i oscillations. All three adrenergic agonists reduced the frequency of the glucose-induced [Ca2+]i oscillations in islets with glucose-induced high frequency [Ca2+]i oscillations, but not in islets, in which glucose induced low frequency [Ca2+]i oscillations. This effect of catecholamines is likely to be mediated via alpha2-adrenoceptors, as supported by the observation that the agonistic effect could be antagonized by yohimbine, a selective alpha2 adrenoceptor antagonist. Thus, whether an individual islet responds to glucose stimulation with high or low frequency [Ca2+]i oscillations appears to be determined at least in part by the adrenergic tone. Furthermore, we could show that glucagon as well as IBMX and forskolin indeed significantly increased the frequency of the glucose-induced [Ca2+]i oscillations. These results support a cAMP mediated regulation and indicate that glucagon release from pancreatic alpha cells acts in a paracrine fashion as a modulator of glucose-induced [Ca2+]i oscillations in mouse pancreatic islets. PMID- 17923117 TI - Improved cytosolic translocation and tumor-killing activity of Tat-shepherdin conjugates mediated by co-treatment with Tat-fused endosome-disruptive HA2 peptide. AB - Tat peptides are useful carriers for delivering biologic molecules into the cell for both functional analysis of intracellular disease-related proteins and treatment of refractory diseases. Most internalized Tat-fused cargos (Tat-cargos) are trapped within the endosome, however, which limits the biologic function of the cargo. In this study, we demonstrated that Tat-fused HA2 peptide (HA2Tat), an endosome disrupted peptide, enhanced the endosome-escape efficiency of Tat cargos. In cells treated with a mixture of fluorescein isothiocyanate-labeled Tat and HA2Tat, widespread fluorescence was observed throughout the cytosol. In addition, this HA2Tat-mediated cytosolic delivery technique led to enhanced cytotoxicity of Tat-fused anti-cancer peptides, specifically shepherdin. Thus, we improved the function of the delivered molecules by co-treating with HA2Tat and propose that this is a useful method for the delivery of therapeutic macromolecules into the cytosol. PMID- 17923118 TI - Stimulation by glutamine and proline of HGF production in hepatic stellate cells. AB - Amino acids regulate cellular functions in a variety of cell types. Most notably, leucine stimulates protein production through the mammalian target of rapamycin (mTOR)-dependent signaling pathway. We investigated the effect of amino acids on hepatocyte growth factor (HGF) production. Treatment with glutamine and proline, as well as leucine, increased HGF levels in the culture medium of a rat hepatic stellate cell clone in a dose-dependent manner. Up-regulation of phosphorylation of 70 kDa ribosomal protein S6 kinase and eukaryotic initiation factor 4E-binding protein 1 was not apparent in the cells after treatment with glutamine or proline. When rats received injections of glutamine or proline, hepatic and circulating HGF levels increased and peaked around 12h after treatment. Glutamine and proline may have the potential to stimulate HGF production but the mechanism underlying this stimulation seems not to be through the mTOR-dependent signaling pathway. PMID- 17923119 TI - Central role of ILT3 in the T suppressor cell cascade. AB - CD8+ T suppressor cells differentiate both in vivo and in vitro upon chronic exposure of responding T cells to allogeneic APC. These Ts are allospecific and exhibit their function interacting directly with priming APC which they render tolerogenic. Tolerogenicity of professional and non-professional human APC, such as dendritic cells and endothelial cells, respectively is due to the upregulation of the inhibitory receptors ILT3 and ILT4. ILT3 signals both intracellularly, inhibiting NF-kappaB activation, and transcription of costimulatory molecules, and extracellularly, inducing anergy and regulatory function in T cells with cognate specificity. Both membrane and soluble ILT3 are proteins with potent immunosuppressive activity which are of importance for treatment of rejection, autoimmunity and cancer. PMID- 17923120 TI - The role of attention in subitizing. AB - The process of rapidly and accurately enumerating small numbers of items without counting, i.e. subitizing, is often believed to rest on parallel preattentive processes. However, the possibility that enumeration of small numbers of items would also require attentional processes has remained an open question. The present study is the first that directly contrasts the preattentive and attentive models of subitizing. We used an inattentional blindness paradigm to manipulate the availability of attentional resources during enumeration. In the inattention condition, the items to be enumerated were presented unexpectedly while participants focused on a line length comparison task. Divided- and full attention conditions were also included. The results showed that only numbers one and two could be enumerated when the effects of attention were minimized. Freeing attentional resources increased the enumeration accuracies considerably, including for number two. The results suggest that even for enumerating small numbers, the attentional demands increase as the number of objects increases. PMID- 17923121 TI - Freezability of rat epididymal sperm induced by raffinose in modified Krebs Ringer bicarbonate (mKRB) based extender solution. AB - The objective of this study was to develop an ideal freezing extender and method for rat epididymal sperm cryopreservation. Epididymal sperm collected from 30 Wistar males was frozen, and experiments were conducted to study its post-thaw characteristics when freezing with raffinose-free buffer or various concentrations of raffinose and egg yolk dissolved in distilled and deionised water, PBS, or modified Krebs-Ringer bicarbonate (mKRB)-based extender. Different concentrations of glycerol, Equex STM, or sodium dodecyl sulfate (SDS) dissolved in either PBS or mKRB containing egg yolk were also tested. Based on the data from these experiments, further experiments tested how different sugars such as raffinose, trehalose, lactose, fructose, and glucose dissolved in mKRB with Equex STM, SDS and egg yolk supplementation affected the post-thaw characteristics of cryopreserved sperm. Cryosurvival of frozen-thawed sperm were judged by microscopic assessment of the sperm motility index (SMI), and acrosome integrity was measured using FITC-PNA staining. Thawed sperm were subjected to 3h of a thermal resistance test. Beneficial effects on the post-thaw survival of sperm were obtained when 0.1M raffinose in mKRB was used with 0.75% Equex STM, 0.05% SDS, and 20% egg yolk. Sperm cryopreserved with this treatment exhibited a higher motility index and maintained greater SMI and acrosome integrity throughout incubation when compared to sperm frozen in various concentrations of other cryoprotectants and trehalose, lactose, fructose, glucose. In conclusion, cryopreservation in an extender solution of raffinose dissolved in mKRB containing Equex STM, SDS and egg yolk greatly enhances the freezability of rat epididymal sperm. PMID- 17923122 TI - The effect of moderate hypothermia in acute ischemic stroke on pericyte migration: an ultrastructural study. AB - Pericytes are essential components of the blood-brain barrier together with endothelial cells and astrocytes. Any disturbance of brain perfusion may result in blood-brain barrier dysfunction due to pericyte migration from the microvascular wall. The neuroprotective influence of hypothermia on ischemic brain injury has been clearly shown in models of both global and focal ischemia. Leakage of plasma proteins contributes to the extension of neuronal injury and hypothermia has a neuroprotective influence during the ischemic insult. This line of thinking impelled us to investigate the possible role of the pericytes in the occurrence of hypothermic protection during cerebral ischemia. In this study, we examined at the ultrastructural level the effect of moderate hypothermia on microvascular pericyte responses using a rat model of permanent middle cerebral artery occlusion. Twenty rats were divided into four groups. Middle cerebral artery occlusion was performed in all rats except the control group (first group), which was used to determine the pericyte morphology under normal conditions. In the second group, pericyte response to irreversible ischemia under normothermic conditions was examined at the end of the first hour. In the third group, pericyte response to hypoxia was examined under normothermic conditions three hours after ischemia. In the fourth group, temporalis muscle temperature was maintained at 27-29 degrees C for 1h after middle cerebral artery occlusion and pericyte response was then examined at the ultrastructural level. In ischemic normothermic conditions at the end of the first hour (Group 2), a separation was observed between pericytes and the basement membrane and this was interpreted as pericyte migration from the microvascular wall. In ischemic normothermic conditions at the end of the third hour (Group 3), basement membrane disorganization and increased space between the basement membranes were seen in addition to the differentiation of second group. In ischemic hypothermic conditions at the end of the first hour (Group 4), pericyte separation or migration from basement membrane were not seen and the blood-brain barrier remained firm. These findings were interpreted by the authors as a possible relationship between pericyte behavior and neural protection during hypothermia. We suggest that hypothermia may delay the pericyte response but not necessarily attenuate it, and should be associated with hypothermic protection. PMID- 17923123 TI - Crystal structure of a family I.3 lipase from Pseudomonas sp. MIS38 in a closed conformation. AB - The crystal structure of a family I.3 lipase from Pseudomonas sp. MIS38 in a closed conformation was determined at 1.5A resolution. This structure highly resembles that of Serratia marcescens LipA in an open conformation, except for the structures of two lids. Lid1 is anchored by a Ca2+ ion (Ca1) in an open conformation, but lacks this Ca1 site and greatly changes its structure and position in a closed conformation. Lid2 forms a helical hairpin in an open conformation, but does not form it and covers the active site in a closed conformation. Based on these results, we discuss on the lid-opening mechanism. PMID- 17923124 TI - A Chinese herbal decoction, Danggui Buxue Tang, activates extracellular signal regulated kinase in cultured T-lymphocytes. AB - Danggui Buxue Tang (DBT) is prepared from Radix Astragali and Radix Angelicae Sinensis. This Chinese herbal decoction has been shown to stimulate the proliferation of T-lymphocytes; however, the action mechanism of this stimulation has not been revealed. In cultured T-lymphocytes, application of DBT markedly induced the cell proliferation, the release of interleukin-2, -6 and -10, as well as the phosphorylation of extracellular signal-regulated kinases (ERK). The pre treatment of ERK inhibitor blocked the DBT-induced immune responses. In addition, the polysaccharide-enriched fraction of DBT showed marked responses on the cultured T-lymphocytes suggesting the important role of DBT polysaccharide in triggering such immune responses. PMID- 17923125 TI - The degree of macromolecular crowding in the cytoplasm and nucleoplasm of mammalian cells is conserved. AB - Macromolecular crowding provides the cytoplasm and the nucleoplasm with strongly viscoelastic properties and renders the diffusion of soluble proteins in both fluids anomalous. Here, we have determined the nanoscale viscoelasticity of the cytoplasm and the nucleoplasm in different mammalian cell lines. In contrast to the cell-specific response on the macroscale the nanoscale viscoelasticity (i.e. the behavior on length scales about 100-fold smaller than the cell size) only showed minor variations between different cell types. Similarly, the associated anomalous diffusion properties varied only slightly. Our results indicate a conserved state of macromolecular crowding in both compartments for a variety of mammalian cells with the cytoplasm being somewhat more crowded than the nucleus. PMID- 17923126 TI - Nucleolar localization of parafibromin is mediated by three nucleolar localization signals. AB - Parafibromin is a putative tumor suppressor encoded by HRPT2 and implicated in parathyroid tumorigenesis. We previously reported a functional bipartite nuclear localization signal (NLS) at residues 125-139. We now demonstrate that parafibromin exhibits nucleolar localization, mediated by three nucleolar localization signals (NoLS) at resides 76-92, 192-194 and 393-409. These NoLS represent clusters of basic amino acids arginine and lysine, similar to those found in other nucleolar proteins, as well as being characteristic of NLSs. While parafibromin's bipartite NLS is the primary determinant of nuclear localization, it does not mediate nucleolar localization. In contrast, the three identified NoLSs play only a minor role in nuclear localization, but are critical for the nucleolar localization of parafibromin. PMID- 17923127 TI - Effect of long-term storage on deoxyribonucleic acid damage and motility of sperm bank donor specimens. AB - The percentage of sperm DNA damage in samples from sperm bank donors was not significantly different (P=.17), whereas the percentage of motile cells was lower (P=.009) after long-term (9-13 years) compared with short-term (1-5 years) storage. Density gradient isolation reduced the difference in sperm motility between the two groups. PMID- 17923128 TI - Effects of non-oral postmenopausal hormone therapy on markers of cardiovascular risk: a systematic review. AB - OBJECTIVE: To review the effects of non-oral administration of postmenopausal hormone therapy (HT) on risk markers for atherosclerotic and venous thromboembolic disease.Non-oral postmenopausal HT appears not to increase venous thromboembolic risk, whereas the effect on coronary heart disease risk is less clear. DESIGN: Systematic review of literature obtained from MEDLINE, EMBASE, and CENTRAL databases from 1980 until and including April 2006. Terms for "postmenopausal hormone therapy" and for "non-oral administration" were combined in the search. SETTING: Randomized clinical trials. PATIENT(S): Postmenopausal women, both healthy and with established cardiovascular disease or specified cardiovascular risk factors INTERVENTION(S): Non-oral HT (e.g., transdermal or intranasal) compared with oral HT or no treatment/placebo. MAIN OUTCOME MEASURE(S): Lipoprotein(a), homocysteine, C-reactive protein (CRP), cell adhesion molecules, markers of endothelial dysfunction, coagulation, and fibrinolysis. RESULT(S): Seventy-two studies investigating either transdermal or intranasal administration were included. For non-oral HT, decreases in lipoprotein(a), cell adhesion molecules, and factor VII generally were significant, resistance to activated protein C (APCr) was slightly increased, and other markers including CRP and homocysteine did not change. Compared with oral HT, changes in CRP and APCr were smaller, changes in cell adhesion molecules and some fibrinolytic parameters tended to be smaller, whereas changes in other factors including lipoprotein(a) and homocysteine did not differ. CONCLUSION(S): Potentially unfavorable changes seen with oral HT on two important markers, CRP and APCr, are substantially smaller with non-oral HT. Non-oral HT has minor effects on the other cardiovascular risk markers studied. Therefore, compared with oral HT, non oral HT appears be safer with respect to atherosclerotic and venous thromboembolic disease risk. PMID- 17923129 TI - Noninvasive metabolomic profiling of embryo culture media using Raman and near infrared spectroscopy correlates with reproductive potential of embryos in women undergoing in vitro fertilization. AB - OBJECTIVE: To determine if metabolomic profiling of embryo culture media correlates with reproductive potential of individual embryos. DESIGN: Prospective study. SETTING: An academic and a private assisted reproduction program; a university research center. PATIENT(S): Women undergoing assisted reproduction treatment. INTERVENTION(S): Sixty-nine spent media samples from 30 patients with known outcome (0 or 100% sustained implantation rates) were individually collected after embryo transfer on day 3 and were evaluated using Raman and/or near-infrared spectroscopy. The spectra obtained from each instrument were separately analyzed using a wavelength selective genetic algorithm to determine regions predictive of pregnancy outcome. Viability indices reflective of reproductive potential were calculated for each sample. To avoid random correlations, a leave-one-out cross-validation was used. Sensitivity and specificity of predicting viability (described as implantation and delivery) were calculated. MAIN OUTCOME MEASURE(S): Metabolomic profile of culture media and embryo viability. RESULT(S): Viability indices calculated by Raman or near infrared spectroscopy were higher for embryos that implanted and resulted in a delivery, compared with those that failed to implant. Raman spectroscopy predicted viability of individual embryos with a sensitivity of 86% and a specificity of 76.5%; near-infrared provided a sensitivity of 75% and a specificity of 83.3%. CONCLUSION(S): Rapid, noninvasive metabolomic profiling of human embryo culture media using Raman or near-infrared spectroscopy combined with bioinformatics correlates with pregnancy outcome. PMID- 17923130 TI - Preconception folic acid treatment affects the microenvironment of the maturing oocyte in humans. AB - OBJECTIVE: To investigate the influence of folic acid supplementation on the follicular fluid concentrations of folate and total homocysteine and their relationship to the diameter of the follicle. DESIGN: Observational study. SETTING: Tertiary referral fertility clinic at the Erasmus MC, University Medical Center, Rotterdam, The Netherlands. PATIENT(S): Thirty-seven women undergoing IVF or intracytoplasmic sperm injection treatment. INTERVENTION(S): No interventions other than routine stimulation treatment and the recommendation of folic acid supplementation. MAIN OUTCOME MEASURE(S): Concentrations of folate and total homocysteine in monofollicular and pooled follicular fluid and the diameter of the follicle. RESULT(S): Folic acid supplementation significantly increased folate and decreased total homocysteine concentrations in pooled follicular fluid. In monofollicular fluid, folate concentrations only were significantly increased in supplemented women. The total homocysteine concentration appeared to be significantly correlated with the diameter of the follicle (r = 0.27). Samples from single follicles were less prone to artifacts in the measurements of the folate and total homocysteine concentration. CONCLUSION(S): Preconception folic acid supplementation significantly alters both folate and total homocysteine concentrations in follicular fluid. The correlation between the diameter of the follicle and total homocysteine concentration in follicular fluid warrants further investigation. PMID- 17923131 TI - Evaluation of anti-Mullerian hormone as a test for the prediction of ovarian reserve. AB - OBJECTIVE: To compare in an integral way the value of the basal serum anti Mullerian hormone (AMH) level with most of the established ovarian reserve tests. DESIGN: Prospective randomized controlled trial. SETTING: Fertility center of a university hospital in the Netherlands. PATIENT(S): One hundred ten patients undergoing their first IVF cycle who were randomized, by a computer-designed four block system, into two groups. INTERVENTION(S): Fifty-six patients underwent a clomiphene citrate challenge test (CCCT), and 54 patients underwent an exogenous FSH ovarian reserve test (EFORT). In all patients, basal AMH, basal FSH, basal inhibin B, antral follicle count (AFC), and basal volume of the ovaries were measured. In all patients, the test was followed by a standard IVF treatment. MAIN OUTCOME MEASURE(S): Ovarian response after ovarian hyperstimulation in an IVF treatment, expressed as the total number of stimulated follicles, retrieved oocytes, and ongoing pregnancies. RESULT(S): The best prediction of ovarian reserve (Y) was seen in a multiple regression prediction model that simultaneously included AFC, inhibin B increment in the EFORT, and basal volume of the ovaries. Univariate logistic regression showed that the best predictors for poor response were AMH, the CCCT, basal FSH, and the AFC. For hyperresponse, univariate logistic regression showed that the best predictor was AFC. Multiple logistic regression analysis did not produce a better model in terms of improving the prediction of poor response or hyperresponse. The best predictors for the prediction of non-pregnancy were the CCCT and the E(2) increment in the EFORT. CONCLUSION(S): Anti-Mullerian hormone is comparable with other commonly used ovarian reserve tests but is probably most applicable in general practice. PMID- 17923132 TI - Insulin resistance in oligomenorrheic infertile women with non-polycystic ovary syndrome. AB - OBJECTIVE: To determine whether infertile oligomenorrheic women are insulin resistant, using an oral glucose tolerance test (OGTT). DESIGN: Retrospective study. SETTING: National Center for Child Health and Development. PATIENT(S): One hundred twenty-seven infertile women with oligomenorrhea (oligomenorrheal group) and 177 infertile eumenorrheic women (normal menstrual group) were recruited. INTERVENTION(S): All women underwent an OGTT (75 g glucose). MAIN OUTCOME MEASURE(S): A homeostasis model assessment of insulin resistance (HOMA-IR), area under the curve (AUC) of insulin after the glucose load, and plasma insulin level at 120 minutes after glucose loading (IRI 120) were used as an index of insulin resistance. RESULT(S): The prevalence of insulin resistance (HOMA-IR >or=1.73) among oligomenorrheic women was 23.8%, which was significantly higher than that of eumenorrheic women, at 14.1%. The glucose AUCs (mean +/- SE) in the oligomenorrheal group (13,609 +/- 259 mg/min/dL) were similar to those for the normal menstrual groups (13,054 +/- 196 mg/min/dL), but the insulin AUCs of the oligomenorrheal group (5333 +/- 376 mU x min/L) were significantly higher than those of the normal menstrual groups (4517 +/- 266 mU x min/L). CONCLUSION(S): The prevalence of insulin resistance assessed using an OGTT was significantly higher among infertile oligomenorrheic women with non-polycystic ovary syndrome than it was among women with normal menstrual cycles. PMID- 17923133 TI - IL-13 pre-treatment of murine peritoneal macrophages increases their anti Toxoplasma gondii activity induced by lipopolysaccharides. AB - Th1 cytokines and microbial lipopolysaccharides (LPS) activate macrophages to produce inflammatory mediators and effector molecules. Althrough Th2 cytokines often have an opposite action to Th1 cytokines and down-modulate the inflammatory response of macrophages, they can induce a distinct alternative activation that is beneficial in host defence. In this study, we report that IL-13 enhances the anti-Toxoplasma activity of LPS-activated murine macrophages. The inhibition of parasite proliferation was not related to reduced Toxoplasma gondii penetration into the cells, nor to the conversion of tachyzoites into bradyzoites. Used alone, IL-13 triggers the polarisation of macrophages towards type 2. However, in LPS-activated macrophages, we show the priming capacity of this cytokine to enhance the expression of inducible nitric oxide synthase (iNOS), a major marker of type 1 macrophages. This effect of IL-13 was not dependent on the activation state of macrophages (resident versus thioglycolate-elicited) or the timing of pre-treatment. We demonstrate a correlation between the enhancement of NO production and upgrading of the microbicidal effectiveness of the macrophages. Thus, both Th2 and Th1 cytokines could activate macrophages to control infections. PMID- 17923134 TI - High-density lipoprotein-cholesterol and risk of stroke and carotid atherosclerosis: a systematic review. AB - BACKGROUND AND PURPOSE: Epidemiological studies have found no relationship between total cholesterol and stroke risk, but little attention has been paid to high-density lipoprotein-cholesterol (HDL-C). METHODS: We performed a systematic PubMed literature search for epidemiological studies that examined the association between HDL-C level and stroke or carotid intima-media thickness (IMT). RESULTS: We identified 18 studies on the relationship between HDL-C and stroke risk and 37 on HDL-C and carotid IMT. Eight of ten prospective cohort studies (n=238,739) and three of eight case-control studies (n=3604 cases, 8220 controls) supported an association between elevated HDL-C level and decreased risk of stroke. Prospective cohort studies reporting on relative risk per unit increase in HDL-C showed an 11-15% decreased stroke risk per 10-mg/dl increase in HDL-C. Of 37 studies on carotid IMT, 31 reported cross-sectional, one longitudinal, and five both cross-sectional and longitudinal associations between HDL-C level and carotid IMT. Of 36 cross-sectional studies (n=51,288), 20 showed an inverse association between HDL-C level and carotid IMT. Of six longitudinal studies (n=20,065), three showed no association, one showed a weak association in a subgroup of white women and two showed a significant inverse relationship between HDL-C level and carotid IMT. Pooled estimates could not be calculated because of the variation in study designs and analysis. CONCLUSIONS: The weight of evidence in the literature supports an inverse association between HDL-C level and stroke or carotid atherosclerosis, but more data are needed to firmly establish this protective effect. PMID- 17923135 TI - Recent development of monolithic stationary phases with emphasis on microscale chromatographic separation. AB - The column technologies play a crucial role in the development of new methods and technologies for the separation of biological samples containing hundreds to thousands compounds. This review focuses on the development of monolithic technology in micro-column formats for biological analysis, especially in capillary liquid chromatography, capillary electrochromatography and microfluidic devices in the past 5 years (2002-2007) since our last review in 2002 on monoliths for HPLC and CEC. The fabrication and functionalization of monoliths were summarized and discussed, with the aim of presenting how monolithic technology has been playing as an attractive tool for improving the power of existing chromatographic separation processes. This review consists of two parts: (i) the recent development in fabrication of monolithic stationary phases from direct synthesis to post-functionalization of the polymer- and silica-based monoliths tailoring the physical/chemical properties of porous monoliths; (ii) the application of monolithic stationary phases for one- and multi-dimensional capillary liquid chromatography, fast separation in capillary electro-driven chromatography, and microfluidic devices. PMID- 17923136 TI - The relationship between the shape of the mental number line and familiarity with numbers in 5- to 9-year old children: evidence for a segmented linear model. AB - This experiment aimed to expand previous findings on the development of mental number representation. We tested the hypothesis that children's familiarity with numbers is directly reflected by the shape of their mental number line. This mental number line was expected to be linear as long as numbers lay within the range of numbers children were familiar with. Five- to 9-year-olds (N=78) estimated the positions of numbers on an external number line and additionally completed a counting assessment mirroring their familiarity with numbers. A segmented regression model consisting of two linear segments described number line estimations significantly better than a logarithmic or a simple linear model. Moreover, the change point between the two linear segments, indicating a change of discriminability between numbers, was significantly correlated with children's familiar number range. Findings are discussed in terms of the accumulator model, assuming a linear mental representation with scalar variability. PMID- 17923137 TI - A simple model of co-evolutionary dynamics caused by epistatic selection. AB - Epistasis is the dependency of the effect of a mutation on the genetic background in which it occurs. Epistasis has been widely documented and implicated in the evolution of species barriers and the evolution of genetic architecture. Here we propose a simple model to formalize the idea that epistasis can also lead to co evolutionary patterns in molecular evolution of interacting genes. This model epistasis is represented by the influence of one gene substitution on the fitness rank of the resident allele at another locus. We assume that increasing or decreasing fitness rank occur equally likely. In simulations we show that this form of epistasis leads to co-evolution in the sense that the length of an adaptive walk between interacting loci is highly correlated. This effect is caused by episodes of elevated rate of evolution in both loci simultaneously. We find that the influence of epistasis on these measures of co-evolutionary dynamics is relatively robust to the details of the model. The main factor influencing the correlation in evolutionary rates is the probability that a substitution will have an epistatic effect, but the strength of epistasis or the asymmetry of the initial fitness ranks of the alleles have only a minor effect. We suggest that covariance in rates of evolution among loci could be used to detect epistasis among loci. PMID- 17923138 TI - Vaccinations in disease models with antibody-dependent enhancement. AB - This paper examines the effects of single-strain vaccine campaigns on the dynamics of an epidemic multistrain model with antibody-dependent enhancement (ADE). ADE is a disease spreading process causing individuals with their secondary infection to be more infectious than during their first infection by a different strain. We follow the two-strain ADE model described in Cummings et al. [D.A.T. Cummings, Doctoral Thesis, Johns Hopkins University, 2004] and Schwartz et al. [I.B. Schwartz, L.B. Shaw, D.A.T. Cummings, L. Billings, M. McCrary, D. Burke, Chaotic desynchronization of multi-strain diseases, Phys. Rev. E, 72:art. no. 066201, 2005]. After describing the model and its steady state solutions, we modify it to include vaccine campaigns and explore if there exists vaccination rates that can eradicate one or more strains of a virus with ADE. PMID- 17923139 TI - Flavones from Struthiola argentea with anthelmintic activity in vitro. AB - Parasitic diseases caused by helminthes lead to significant health hazards to animals resulting in enormous economic impact. While a number of anthelmintics are currently available, all are encountering resistance and ones with a mode of action are needed. We report herein bioassay-guided isolation of three anthelmintic flavones 1-3, including the flavone, 5,6,2',5',6'-pentamethoxy-3',4' methylenedioxyflavone (3) from the methanol extract of Struthiola argentea (Thymelaeaceae). The structure of 3 was elucidated by analysis of its 1D and 2D NMR and MS data. The two major flavones produced by this plant were also isolated and identified as yuankanin (4) and amentoflavone (5). A number of flavones related to the compounds isolated from S. argentea were acquired and tested to ascertain structure activity relationships. The isolation, structure, anthelmintic activity and structure activity relationships of the flavones are described. Compound 3 exhibited the most potent in vitro activity with 90% inhibition of larval motility at 3.1 microg/mL and compound 15 showed modest in vivo activity. PMID- 17923140 TI - Exploring environmental determinants of physical activity--the road to the future is always under construction. PMID- 17923141 TI - The role of economic and educational factors in asthma: evidence from the Portuguese health survey. AB - OBJECTIVE: To evaluate the importance of economic and educational factors in determining the risk of asthma in adults. STUDY DESIGN AND METHODS: This was a cross-sectional study in a representative sample of Portuguese adults (20,977 females and 18,663 males) from the main regions of mainland Portugal. Participants were categorized according to years of education (< or = 4, 5-9, 10 12 and >12 years) and income (< or = 314, 315-547, 548-815 and >815Euros/month). Information on asthma was based on answers to the following question: 'Have you had asthma in the previous year?' Logistic regression models were fitted to estimate the magnitude of the association between asthma and education/income, adjusting for confounders (body mass index, age, smoking habits, physical activity, area of residence, number of household members and income/education). RESULTS: In females and males, the odds of having asthma decreased with increasing income (P-value for trend <0.001), with odds ratios of 0.52 [95% confidence intervals (CI) 0.41-0.66] and 0.55 (95%CI 0.44-0.68) for those with a monthly income >815 Euros compared with those with a monthly income 314 Euros, respectively. For both genders, the odds of having asthma were not significantly associated with years of education. CONCLUSION: A significant positive association between per capita national income and asthma reinforces that public policy should stress the importance of economic growth as a means for preventing asthma occurrence and improving quality of life. PMID- 17923143 TI - Versatility of AAV vectors for retinal gene transfer. AB - Gene therapy represents a promising therapeutic option for many inherited and acquired retinal diseases. Recombinant adeno-associated viral vectors (AAV) are the most efficient tools to transfer genes in vivo to the retina. The recent identification of dozens of novel AAV serotypes enormously expands on the versatility of AAV as vector system for in vivo somatic gene transfer. The results from the forthcoming trials with AAV in the retina of patients with Leber Congenital Amaurosis will be critical for the rapid development of AAV-based therapeutics for retinal diseases. PMID- 17923142 TI - SANS (USH1G) expression in developing and mature mammalian retina. AB - The human Usher syndrome (USH) is the most common form of combined deaf blindness. Usher type I (USH1), the most severe form, is characterized by profound congenital deafness, constant vestibular dysfunction and prepubertal onset of retinitis pigmentosa. Five corresponding genes of the six USH1 genes have been cloned so far. The USH1G gene encodes the SANS (scaffold protein containing ankyrin repeats and SAM domain) protein which consists of protein motifs known to mediate protein-protein interactions. Recent studies indicated SANS function as a scaffold protein in the protein interactome related to USH. Here, we generated specific antibodies for SANS protein expression analyses. Our study revealed SANS protein expression in NIH3T3 fibroblasts, murine tissues containing ciliated cells and in mature and developing mammalian retinas. In mature retinas, SANS was localized in inner and outer plexiform retinal layers, and in the photoreceptor cell layer. Subcellular fractionations, tangential cryosections and immunocytochemistry revealed SANS in synaptic terminals, cell cell adhesions of the outer limiting membrane and ciliary apparati of photoreceptor cells. Analyses of postnatal developmental stages of murine retinas demonstrated SANS localization in differentiating ciliary apparati and in fully developed cilia, synapses, and cell-cell adhesions of photoreceptor cells. Present data provide evidence that SANS functions as a scaffold protein in USH protein networks during ciliogenesis, at the mature ciliary apparatus, the ribbon synapse and the cell-cell adhesion of mammalian photoreceptor cells. Defects of SANS may cause dysfunction of the entire network leading to retinal degeneration, the ocular symptom characteristic for USH patients. PMID- 17923144 TI - Observers are consistent when rating image conspicuity. AB - Human perception of an image's conspicuity depends on the stimulus itself and the observer's semantic interpretation. We investigated the relative contribution of the former, sensory-driven, component. Participants viewed sequences of images from five different classes-fractals, overhead satellite imagery, grayscale and colored natural scenes, and magazine covers-and graded each numerically according to its perceived conspicuity. We found significant consistency in this rating within and between observers for all image categories. In a subsequent recognition memory test, performance was significantly above chance for all categories, with the weakest memory for satellite imagery, and reaching near ceiling for magazine covers. When repeating the experiment after one year, ratings remained consistent within each observer and category, despite the absence of explicit scene memory. Our findings suggest that the rating of image conspicuity is driven by image-immanent, sensory factors common to all observers. PMID- 17923145 TI - Effects of light on sediment nutrient flux and water column nutrient stoichiometry in a shallow lake. AB - The effects of light and temperature on nutrient cycling (silica (Si), nitrogen (N) and phosphorus (P)) between sediments and water in a shallow eutrophic lake (Loch Leven, Scotland), and consequent effects on water column nutrient stoichiometry, were assessed using a series of intact sediment core incubation experiments. Estimates of actual seasonal dark and light P-fluxes were assessed using 24-h incubations. Sediment-P uptake was observed in spring (7 degrees C) and release in autumn (12 degrees C) and summer (17 degrees C), with the highest release rates ( approximately 17 mgPO4-Pm(-2) sediment surface area d(-1)) occurring in summer. In a longer (21-day) experiment in which the effects of light (light (n=6) and dark (n=6)) and temperature (five 4-day cycles to represent: 7 degrees C-->13 degrees C-->23 degrees C-->13 degrees C-->7 degrees C) on water column nutrient concentrations were assessed, PO(4-)-P, total P (TP), SiO2 and total silica (TSi) concentrations in the water column were all significantly higher under dark conditions (ANOVA, alpha=0.05). NH4-N (ammonium N) water column concentrations were observed to be higher under dark conditions at low temperatures and higher under light conditions following a high temperature (23 degrees C) treatment. No significant light effects were observed for water column total N (TN) concentration. Flux estimates for all nutrients measured are given. In terms of water column nutrient stoichiometry, TN:TP ratio was significantly higher under light conditions, TSi:TN was significantly lower under light conditions, and TSi:TP did not vary significantly between the dark and light treatments. The main processes acting to regulate diffusive nutrient release appeared to be photosynthetic elevation of bottom water pH and dissolved oxygen concentration (both significantly higher under light conditions) and direct microalgal sequestration. Thus, a feedback mechanism exists in recovering shallow lakes where benthic microalgae can affect the stoichiometry (to favour P/Si limitation) of the plankton, and also of the main source of nutrients back to the sediments via the disproportionate regulation of sediment P, Si and N release. PMID- 17923146 TI - Electrochemical oxidation of table olive processing wastewater over boron-doped diamond electrodes: treatment optimization by factorial design. AB - The electrochemical treatment of an effluent from edible olive processing over boron-doped diamond electrodes was investigated. The effect of operating conditions, such as initial organic loading (from 1340 to 5370 mg/L chemical oxygen demand (COD)), reaction time (from 30 to 120 min), current intensity (from 5 to 14 A), initial pH (from 3 to 7) and the use of 500 mg/L H2O2 as an additional oxidant, on treatment efficiency was assessed implementing a factorial experimental design. Of the five parameters tested, the first three had a considerable effect on COD and total phenols removal, while the other two were statistically insignificant. In most cases, high levels of phenols degradation and decolorization were achieved followed by moderate mineralization. The analysis was repeated at more intense conditions, i.e., initial COD up to 10,000 mg/L, reaction times up to 240 min and current up to 30 A; at this level, the effect of treatment time and applied current was far more important than the starting COD concentration. Treatment for 14 h at optimal conditions (30 A and an initial loading of about 10,000 mg/L) led to 73% COD removal with a zero-order kinetic constant of 8.5mg/(L min) and an energy consumption efficiency of 16.3 g COD/(m3 A h). PMID- 17923147 TI - The carbon-sequestration potential of municipal wastewater treatment. AB - The lack of proper wastewater treatment results in production of CO(2) and CH(4) without the opportunity for carbon sequestration and energy recovery, with deleterious effects for global warming. Without extending wastewater treatment to all urban areas worldwide, CO(2) and CH(4) emissions associated with wastewater discharges could reach the equivalent of 1.91 x 10(5) t(CO2)d(-1) in 2025, with even more dramatic impact in the short-term. The carbon sequestration benefits of wastewater treatment have enormous potential, which adds an energy conservation incentive to upgrading existing facilities to complete wastewater treatment. The potential greenhouse gases discharges which can be converted to a net equivalent CO(2) credit can be as large as 1.91 x 10(5) t(CO2)d(-1) in 2025 by 2025. Biomass sequestration and biogas conversion energy recovery are the two main strategies for carbon sequestration and emission offset, respectively. The greatest potential for improvement is outside Europe and North America, which have largely completed treatment plant construction. Europe and North America can partially offset their CO(2) emissions and receive benefits through the carbon emission trading system, as established by the Kyoto protocol, by extending existing technologies or subsidizing wastewater treatment plant construction in urban areas lacking treatment. This strategy can help mitigate global warming, in addition to providing a sustainable solution for extending the health, environmental, and humanitarian benefits of proper sanitation. PMID- 17923148 TI - Effects of SOM, surfactant and pH on the sorption-desorption and mobility of prometryne in soils. AB - Sorption and desorption of the herbicide prometryne in two types of soil subjected to the changes of pH and soil organic matter and surfactant were investigated. The sorption and desorption isotherms were expressed by the Freundlich equation. Freundlich K(f) and n values indicate that soil organic matter was the major factor affecting prometryne behavior in the test soils. We also quantified the prometryne sorption and desorption behavior in soils, which arose from the application of Triton X-100 (TX100), a nonionic surfactant and change in pH. Application of TX100 led to a general decrease in prometryne sorption to the soils and an increase in desorption from the soils when applied in dosages of the critical micella concentration (CMC) 0.5, 1 and 2. At the concentration below the CMC, the non-ionic surfactant showed a tendency to decrease prometryne sorption and desorption. It appeared that TX100 dosages above CMC were required to effectively mobilize prometryne. Results indicate that the maximum prometryne sorption and minimum prometryne desorption in soils were achieved when the solution pH was near its pK(a). Finally, the influence of TX100 on the mobility of prometryne in soils using soil thin-layer chromatography was examined. PMID- 17923149 TI - Excess lead alters growth, metabolism and translocation of certain nutrients in radish. AB - To elucidate the deleterious effects of excess lead on radish (Raphanus sativus) cv. Jaunpuri plants were grown in refined sand in complete nutrient solution for 30 days. On the 31st day lead nitrate was superimposed at 0.1 and 0.5mM to radish for 65 days. A set of plants in complete nutrient solution was maintained as control for the same period without lead. Excess Pb at 0.5mM showed growth depression with interveinal chlorosis on young leaves at apex. Excess Pb reduced the fresh and dry weight pronouncedly at d 65. Lead accumulation reduced the concentration of chlorophyll, iron, sulphur (in tops), Hill reaction activity and catalase activity whereas increased the concentration of phosphorus, sulphur (in roots) and activity of peroxidase, acid phosphatase and ribonuclease in leaves of radish. PMID- 17923150 TI - Connecting local environmental sequences to global climate patterns: evidence from the hominin-bearing Hadar Formation, Ethiopia. AB - Central to the debate surrounding global climate change and Plio-Pleistocene hominin evolution is the degree to which orbital-scale climate patterns influence low-latitude continental ecosystems and how these influences can be distinguished from regional volcano-tectonic events and local environmental effects. The Pliocene Hadar Formation of Ethiopia preserves a record of hominin paleoenvironments from roughly 3.5 to 2.2 Ma at a temporal resolution relevant to evolutionary change within hominins and other taxa. This study integrates the high-resolution sedimentological and paleontological records at Hadar with climate proxies such as marine core isotope, dust, and sapropel records. Consistent cycling observed both between and within fluvial and lacustrine depositional environments prior to 2.9 Ma at Hadar appears to be predominantly climatic in nature. In contrast a significant change in depositional facies after 2.9 Ma to sequences dominated by conglomerate cut-and-fill cycles indicates a strong tectonic signature related to regional developments in the Main Ethiopian Rift. While specific events seen in marine proxy records may have parallels in the Hadar environmental archive, their overall patterns of high versus low variability may be even more relevant. For example, periods of relatively high amplitude climate oscillations between 3.15 and 2.95 Ma may be linked to noted size-related morphological changes within the Hadar Australopithecus afarensis lineage and a significant increase in more arid-adapted bovid taxa. Increased aridity in East Africa during this period is also indicated by peaks in eolian dust in the marine core record. Conversely, the dominant lacustrine phase at Hadar ca. 3.3 Ma coincides with the least variable period in several climate proxy records, including marine core foraminifera delta(18)O values and eolian dust concentration. This phase is also coeval with low insolation variability and a very distinct and significant long-term period of low dust percentage in circum Africa marine cores. PMID- 17923151 TI - Investigating early hominin dispersal patterns: developing a framework for climate data integration. AB - An investigation using the Stepping Out model of early hominin dispersal out of Africa is presented here. The late arrival of early hominins into Europe, as deduced from the fossil record, is shown to be consistent with poor ability of these hominins to survive in the Eurasian landscape. The present study also extends the understanding of modelling results from the original study by Mithen and Reed (2002. Stepping out: a computer simulation of hominid dispersal from Africa. J. Hum. Evol. 43, 433-462). The representation of climate and vegetation patterns has been improved through the use of climate model output. This study demonstrates that interpretative confidence may be strengthened, and new insights gained when climate models and hominin dispersal models are integrated. PMID- 17923152 TI - Simulation of spontaneous action potentials of cardiomyocytes in pulmonary veins of rabbits. AB - Atrial fibrillation is the most prevalent arrhythmia, but the mechanisms by which it develops are not clear. Recently, over 90% of paroxysmal atrial fibrillation was found to be located inside the main pulmonary veins (PVs). We found that single cardiac myocytes isolated from the main PVs of rabbits generate spontaneous action potentials (SAP). We therefore assayed the electrical characteristics of these cardiomyocytes. Among the diverse ionic currents identified were INa, ICa,L, IK1, IKr, IKs, Ito, IKsus, Incx, Ipump, IKH and ICl,Ca. In contrast, IK1 was minimal, IKs could be detected only in the presence of 10 microM forskolin, and we were unable to detect If and ICa,T, the most important currents for pacemaking activity in sinoatrial node cells. To identify the main cause of SAP, we developed a model that can explain the electrical properties of these cardiomyocytes. After reconstructing the ionic currents based on experimental observations, we were able to use our model to successfully reconstruct the characteristics of the SAP of PV cardiomyocytes. The simulation showed that the major currents contributing to pacemaking depolarization were ICaL, IKr, a background current and Na+-K+ pump current. Deactivation kinetics of IKr was one of the major determinants of the rate of pacemaking depolarization. The steady state inactivation of Ito was shifted to the negative voltage and the activity of Ito was minimal in the range of the SAP. The major currents for the repolarization were IKr and Ipump. The amplitude of most currents in these cardiac myocytes was small and no currents did not exceed 30 pA during the SAP, indicating that slight activation of other inward or outward currents will have profound effects on the SAP. To our knowledge, this report is the first to show the simulation of SAP of PV cardiomyocytes. This model may help to study on the electrophysiological basis of paroxysmal atrial fibrillation originating from PVs. PMID- 17923153 TI - Quantitative determination of radio-opacity: equivalence of digital and film X ray systems. AB - OBJECTIVES: To evaluate the equivalence of a digital X-ray system (DenOptix) to conventional X-ray film in terms of the measured radio-opacity of known filled resin materials and the suitability of attenuation coefficient for radio-opacity determination. METHODS: Discs of five thicknesses (0.5-2.5mm) and step-wedges of each of three composite materials of nominal aluminum-equivalence of 50%, 200% and 450% were used. X-ray images of a set of discs (or step-wedge), an aluminum step-wedge, and a lead block were taken at 65 kV and 10 mA at a focus-film distance of 400 mm for 0.15s and 1.6s using an X-ray film or imaging plate. Radio opacity was determined as equivalent aluminum thickness and attenuation coefficient. The logarithm of the individual optical density or gray scale value, corrected for background, was plotted against thickness, and the attenuation coefficient determined from the slope. The method of ISO 4049 was used for equivalent aluminum thickness. RESULTS: The equivalent aluminum thickness method is not suitable for materials of low radio-opacity, while the attenuation coefficient method could be used for all without difficulty. The digital system gave attenuation coefficients of greater precision than did film, but the use of automatic gain control (AGC) distorted the outcome unusably. CONCLUSION: Attenuation coefficient is a more precise and generally applicable approach to the determination of radio-opacity. The digital system was equivalent to film but with less noise. The use of AGC is inappropriate for such determinations. PMID- 17923154 TI - A multifunctional and reversibly polymerizable carrier for efficient siRNA delivery. AB - In this study a multifunctional carrier (MFC), 1,4,7-triazanonylimino-bis[N (oleicyl-cysteinyl-histinyl)-1-aminoethyl)propionamide] (THCO), containing protonatable amines of different pK(a)s, polymerizable cysteine residues and hydrophobic groups, was designed, synthesized and evaluated for efficient small interfering RNAs (siRNA) delivery. THCO showed pH-sensitive cellular membrane disruption at the endosomal-lysosomal pH to facilitate intracellular siRNA delivery. THCO formed stable and compact nanoparticles with siRNA through charge complexation, hydrophobic condensation and reversible polymerization. The THCO/siRNA nanoparticles were readily modified with PEG-Mal by reacting with remaining thiol groups at the surface. The siRNA delivery efficiency of THCO was comparable to that of Transfast, much higher than that of N-(2,3-dioleoyloxy-1 propyl)trimethylammonium methyl sulphate (DOTAP) in serum-free medium. PEGylated THCO/siRNA nanoparticles resulted in higher transfection efficiency than those of Transfast and DOTAP in the presence of serum. This study demonstrated that the MFC-THCO is promising for efficient siRNA delivery. PMID- 17923156 TI - The global eradication of poliomyelitis: progress and problems. AB - Encouraged by the success of the global smallpox eradication certified in 1980, the global poliomyelitis eradication program was launched in 1988 by the World Health Organization (WHO). In addition to routine polio immunization included in the Expanded Program of Immunization (EPI), two major activities were planned: mass polio vaccination campaigns and surveillance of all cases of acute flaccid paralysis. In 2000, the disease had been eliminated from most countries in the world. However, as of 2002, the community acceptance of vaccination was endangered in some countries by rumors about assumed adverse effects of oral polio vaccine. The rejection of polio immunization provided a worrying resurgence of polio in Northern Nigeria, followed by re-infection of 21 countries, whereas resurgence of the disease also was observed in Northern India. Supplementary vaccination activities were resumed, additional resources were mobilized and, in 2007, most re-infected countries became polio-free again. Today, polio remains endemic in only four countries. The goal of global polio eradication has now been set at 2010, but doubts have been expressed about the feasibility of its achievement. PMID- 17923155 TI - Breast cancer resistance protein: mediating the trans-placental transfer of glyburide across the human placenta. AB - Members of the ATP-binding cassette (ABC) efflux transporter family, including P glycoprotein (PGP), the multidrug resistance-associated proteins (MRPs) and the breast cancer resistance protein (BCRP) have been shown to be highly expressed in the human placenta. Recent studies documented that the oral hypoglycemic glyburide does not cross the human placenta to an appreciable extent. Furthermore, the trans-placental transfer of glyburide has been shown not to be affected by either the presence of PGP inhibitor, verapamil or MRP inhibitor, indomethacin. Therefore, our objective was to identify other human placental ABC transporters potentially involved in limiting the trans-placental transfer of glyburide to the fetus. [(3)H]-glyburide transport was examined in brush border human placental vesicles in the presence or absence of specific inhibitors. Prepared vesicles were 70% oriented right-side-out and demonstrated 25-27 fold enrichment as compared to whole placenta. Functional studies demonstrated significant increases in the intra-vesicular accumulation of [(3)H]-glyburide in vesicles treated with the BCRP inhibitor, novobiocin. In contrast, PGP inhibition as well as MRP inhibition did not affect [(3)H]-glyburide accumulation. This is the first evidence to clearly indicate that glyburide is preferentially transported by BCRP, in the brush border of the human placenta. Our study also indicates that BCRP likely effluxes substrates in the fetal to maternal direction in the human placenta. PMID- 17923157 TI - Catalytic turnover dependent modification of the Pseudomonas aeruginosa heme oxygenase (pa-HO) by 5,6-O-isopropyledine-2-O-allyl-ascorbic acid. AB - Heme oxygenase (HO) catalyzes the NADPH dependent conversion of heme to biliverdin with the release of iron and CO via three successive oxygenation steps. The oxidation of heme in the presence of alternate reductants, such as ascorbic acid, has been used extensively to characterize the mechanism of oxygen activation in HO without altering the chemistry of the reaction. NADPH-dependent cytochrome P450 reductase (CPR) and ascorbic acid mediated reactions are mechanistically very similar, in that both use molecular oxygen to initiate the reaction. In the present manuscript, we report on an ascorbic acid derivative, 5,6-O-isopropyledine-2-O-allyl-ascorbic acid, that during catalysis partitions the reaction between the conversion of heme to biliverdin, and an alternate pathway that traps the verdoheme intermediate as a result of protein modification. We propose that following activation of 5,6-O-isopropyledine-2-O allyl-ascorbic acid to the cation radical, protein modification results via alkylation of an active site nucleophile (Asp or Glu), trapping the Fe(III) verdoheme intermediate. The potential site of the modification and the relevance to the mechanism of Fe(III)-verdoheme conversion to biliverdin is discussed. PMID- 17923158 TI - Construction and implantation of a microinfusion system for sustained delivery of neuroactive agents. AB - Experimental protocols used for chronic infusion of neuroactive agents within regions of the brain often utilize a mini-osmotic pump system. Agents are commonly delivered via a stainless steel cannula with a diameter of 0.30 mm or greater. Systems utilizing a cannula of this caliber may impose trauma to the area of interest resulting in architectural damage, thereby compromising structural integrity and normal functioning. As neuroscience inquiry becomes more sophisticated, investigation of brain structures and circuitry requires improved levels of accuracy and higher resolution. We have developed a method for the preparation and implantation of a chronic infusion system within the brain utilizing a borosilicate microcannula with a tip diameter of 50 microm. This technique reduces damage to the local environment and diminishes reactive gliosis at the site of infusion. The configuration of the microinfusion system is also able to conform to the surface of the animal's skull, precluding the need for large cranial pedestals, and thus facilitating closure of the scalp incision and reducing the risk of infection. We demonstrate reliable sustained delivery of a dye having a representative molecular weight using an in vitro model and in vivo studies in rats. PMID- 17923159 TI - Increased water temperature renders single-housed C57BL/6J mice susceptible to antidepressant treatment in the forced swim test. AB - To investigate genotype x environment interactions in the forced swim test, we tested the influence of water temperature (20 degrees C, 25 degrees C, 30 degrees C) on floating behaviour in single-housed male C57BL/6J and BALB/c mice. We observed a contrasting relationship between floating and water temperature between the two strains, with C57BL/6J floating more and BALB/c floating less with increasing water temperature, independent of the lightening conditions and the time point of testing during the animals' circadian rhythm. Both strains showed an inverse relationship between plasma corticosterone concentration and water temperature, indicating that the differences in stress coping are unrelated to different perception of the aversive encounter. Treatment with desipramine (20mg/kg, i.p.) caused a reduction in immobility time in C57BL/6J mice if the animals were tested at 30 degrees C water temperature, with no effect at 25 degrees C and no effects on forced swim stress-induced corticosterone secretion. The same treatment failed to affect floating behaviour in BALB/c at any temperature, but caused a decrease in plasma corticosterone levels. Taken together we demonstrate that an increase in water temperature in the forced swim test exerts opposite effects on floating behaviour in C57BL/6J and BALB/c and renders single-housed C57BL/6J mice, but not BALB/c mice, susceptible to antidepressant-like behavioral effects of desipramine. PMID- 17923160 TI - Oxidative and modulatory effects of trace metals on metabolism of polycyclic aromatic hydrocarbons in the Antarctic fish Trematomus bernacchii. AB - Biological interactions between various classes of pollutants are of great relevance for the Antarctic marine environment, where the naturally elevated bioavailability of metals like cadmium might indirectly influence sensitivity of endemic organisms toward other environmental pollutants, e.g. polycyclic aromatic hydrocarbons (PAHs). To further investigate reciprocal effects of different chemicals, the fish Trematomus bernacchii was exposed to trace metals (Cd, Cu, Hg, Ni, Pb) and benzo[a]pyrene (BaP, as a model PAH), dosed alone and in combinations. Co-exposures revealed that BaP did not influence the accumulation of metals, while these elements caused significant changes on tissue levels of the PAH. The marked EROD induction caused by BaP was completely suppressed by co exposure with Cd and Cu, but no effects were observed with Ni, Hg and Pb. Similar results were confirmed at the protein level by Western blot analyses while CYP1A1 mRNA levels were reduced only during Cd co-exposures. Clear evidence of oxidative perturbations was observed in fish co-treated with Cd and BaP and the reduced capability to absorb peroxyl and hydroxyl radicals suggested some oxidative pathways by which this element might indirectly modulate the biotransformation efficiency of Cytochrome P450. Partly different and post-transcriptional mechanisms of action could be hypothesized for Cu, while moderate oxidative effects of Hg, Ni and Pb during co-exposures would confirm their limited influence on metabolism of PAHs. In general, the overall results revealed a complex pathway of interactions between different chemicals during co-exposures and the importance of oxidative status in modulating induction and expression of CYP1A1. PMID- 17923161 TI - Four-dimensional multislice helical CT of the lung: qualitative comparison of retrospectively gated and static images in an ex-vivo system. AB - PURPOSE: To analyse the image quality of retrospectively gated helical CT using controlled respiratory motion of porcine lung explants. MATERIALS AND METHODS: Five porcine lungs were examined inside a chest phantom. A silicone membrane was rhythmically inflated and deflated to simulate diaphragmatic respiration. Dynamic images (regular respiration at 8/min) and static scans (w/o respiration) at 0/25/50/75 and 100% of maximum inspiration were acquired with a 40-row detector CT scanner (rotation time 1s, pitch 0.1). Image quality on multi-planar reformations was evaluated by two observers. Partial projection artifacts, stepladder-artifacts and noise were compared for upper, middle and lower parts of the lung and different respiratory phases (scores 0-3 for absent, minimal, moderate and diagnostically relevant artifacts). RESULTS: Partial projection effects were limited to dynamic scans (mean score 1.33). Stepladder artifacts predominated in dynamic series compared to static series (mean score 0.55 versus 0.1; p<0.001). Image noise was not related to lung motion (mean scores 0.68 0.81). All artifacts predominated close to the diaphragm compared to the upper and middle parts of the lung (p<0.001 to p=0.02, respectively). Partial projection and stepladder artifacts were less in end-inspiration and end expiration than within the respiration (p<0.001 and p=0.17, respectively). Diagnostically relevant artifacts were noted 9 times (9/9 close to diaphragm, 7/9 partial-projection). CONCLUSIONS: Even in ideal realistic conditions, helical 4D CT produced tolerable artifacts which could be overcome by radiologists. PMID- 17923162 TI - Sparing the parotid glands and surgically transferred submandibular gland with helical tomotherapy in post-operative radiation of head and neck cancer: a planning study. AB - BACKGROUND AND PURPOSE: To evaluate the feasibility of sparing the parotid glands and surgically transferred submandibular gland (SMG) by intensity modulated radiotherapy (IMRT) in post-operative cases of head and neck cancer (HNC). MATERIALS AND METHODS: Ten patients (larynx-2, base of tongue-4, tonsil-3, and unknown primary-1; pathologic stages III-IV) who underwent SMG transfers on the side of N0 neck along with definitive surgery were selected for this study. IMRT planning was done retrospectively using helical tomotherapy approach. Planning objective was to deliver 60 Gy to PTV1 and 54 Gy to PTV2 while maintaining the mean dose to the total parotid volume (TPV) and SMG less than 26 Gy. RESULTS: The mean dose (+/-SD) to the TPV and SMG were 25+/-0.6 Gy and 23+/-1.9 Gy, respectively. The D(95) for PTV1 and PTV2 were 59.9+/-0.1 Gy and 54.9+/-0.3 Gy, respectively, satisfying our planning goal for PTV coverage. The D(99) for PTV1 and PTV2 were 58.2+/-0.7 Gy and 49.5+/-2.2 Gy, respectively, showing that sparing the salivary glands did not result in underdosing of the PTVs. CONCLUSIONS: By combining the gland transfer and IMRT, the mean dose to TPV and transferred SMG could be reduced to less than 26 Gy in post-operative patients of HNC. PMID- 17923163 TI - Concomitant chemoirradiation with vinorelbine and gefitinib induces additive effect in head and neck squamous cell carcinoma cell lines in vitro. AB - BACKGROUND AND PURPOSE: The study was performed to measure the radiosensitizing effect of vinorelbine together with gefitinib in head and neck squamous cell carcinoma (HNSCC) cell lines in vitro. MATERIALS AND METHODS: Three recently established HNSCC cell lines, originating from larynx and oral cavity tumors, were tested in this study. Vinorelbine concentration of 0.5 nM was used, corresponding to the IC(70) value of cell lines. Gefitinib concentrations of 0.10 0.35 microM were used, corresponding to IC(70)- and IC(50)-values of each cell line, causing 30% and 50% inhibition in clonogenic survival, respectively. Vinorelbine was added to the medium and the cells were plated in 96-well culture plates in this solution. 24h later the cells were irradiated in plates with 4 MeV photons generated by a linear accelerator, producing radiation doses 0.75-7.5 Gy. Immediately after irradiation the desired concentrations of gefitinib were added, whereafter the plates were incubated at 37 degrees C with 5% CO(2). After four weeks, the number of wells containing coherent living colonies, consisting of 32 cells or more, was counted. The plating efficiency was calculated and the fraction survival data were fitted to the linear quadratic model, F=exp[ (alphaD+betaD(2))]. The area under the survival curve (AUC) value was obtained with numerical integration. ErbB receptor expression of the HNSCC cell lines was analyzed by Western blotting. RESULTS: The growth-inhibitory effect of simultaneous vinorelbine and gefitinib concomitant with radiation was supra additive in cell line UT-SCC-33 and additive in cell lines UT-SCC-19A and -34. CONCLUSIONS: HNSCC is in vitro constantly sensitive to the combination of vinorelbine and gefitinib, which have together an additive effect in concomitant use with irradiation. Further studies are warranted to evaluate the concomitant use of vinorelbine and gefitinib with irradiation in clinical studies. In addition, both drugs are available in an oral formulation allowing effortless administration schedules. PMID- 17923165 TI - Interferon-gamma exacerbates liver damage, the hepatic progenitor cell response and fibrosis in a mouse model of chronic liver injury. AB - BACKGROUND/AIMS: Several previous studies have suggested that interferon gamma (IFNgamma) may play a key role during hepatic progenitor cell (HPC) mediated liver regeneration. However to date, no studies have directly tested the ability of IFNgamma to mediate the HPC response in an in vivo model. METHODS/RESULTS: Administration of IFNgamma to mice receiving a choline deficient, ethionine (CDE) supplemented diet to induce chronic injury resulted in an augmented HPC response. This was accompanied by increased inflammation, altered cytokine expression and hepatic fibrosis. Serum alanine aminotransferase activity, hepatocyte apoptosis and Bak staining were significantly increased in IFNgamma-treated, CDE-fed mice, demonstrating that liver damage was exacerbated in these animals. Administration of IFNgamma to control diet fed mice did not induce liver damage, however it did stimulate hepatic inflammation. CONCLUSIONS: Our results suggest that IFNgamma increases the HPC response to injury by stimulating hepatic inflammation and aggravating liver damage. This is accompanied by an increase in hepatic fibrogenesis, supporting previous reports which suggest that the HPC response may drive fibrogenesis during chronic liver injury. PMID- 17923164 TI - Structure of orbitofrontal cortex and its longitudinal course in cancer-related post-traumatic stress disorder. AB - The neurobiological basis of cancer-related post-traumatic stress disorder (PTSD) has never been studied. We investigated brain structural alterations and the longitudinal courses in patients with cancer-related PTSD. Baseline scans using magnetic resonance imaging were performed in 14 cancer survivors with PTSD, 100 without PTSD, and 70 healthy subjects. Follow-up scans were performed 2 years later in 76 cancer survivors (PTSD, n=9; non-PTSD, n=67). Using voxel-based morphometry, the gray matter volume (GMV) of the cancer survivors with PTSD was compared with the GMVs of those without PTSD and of the healthy subjects. The effects of the interactions between the diagnosis and the timing of the GMV measurements were examined. The GMV of the right orbitofrontal cortex (OFC) was significantly smaller in cancer survivors with PTSD than in those without PTSD or healthy subjects. The interaction between the diagnosis and the timing of the right OFC's GMV measurement was not significant. The OFC, which is thought to be involved in the extinction of fear conditioning and the retrieval of emotional memory, might play an important role in the pathophysiology of PTSD. Moreover, the OFC's GMV may remain constant after the development of cancer-related PTSD. PMID- 17923167 TI - Effect of ABA-beta-D-glucopyranosyl ester and activity of ABA-beta-D-glucosidase in Arabidopsis thaliana. AB - Exogenously applied ABA-beta-D-glucopyranosyl ester (ABA-GE) inhibited hypocotyl growth of Arabidopsis seedlings at concentrations greater than 0.3 micromol/L, and the concentration for 50% inhibition of hypocotyl growth was 1.8 micromol/L. ABA-beta-D-glucosidase activity in Arabidopsis seedlings was 17 nmol/mg protein/mim and increased by exogenously applied ABA-GE. The pH optimum of this enzyme in crude extract of Arabidopsis seedlings was 6.0 for the assay in the ABA GE to ABA direction and its K(m) value for ABA-GE (pH 6.0) was 0.41 mmol/L. These results suggests that exogenously applied ABA-GE may be absorbed by roots and hydrolyzed by ABA-beta-D-glucosidase and librated free ABA may induce growth inhibition in Arabidopsis hypocotyls. PMID- 17923166 TI - Heterogeneity of CK2 phosphorylation sites in the NS5A protein of different hepatitis C virus genotypes. AB - BACKGROUND/AIMS: The hepatitis C virus NS5A protein is phosphorylated by several cellular kinases, including casein kinase 2 (CK2). Little is known about CK2 phosphorylation of NS5A from different HCV genotypes and clinical isolates. METHODS: NS5A from patients with HCV-1a (24 cases), HCV-1b (9) or HCV-3 (16) was analyzed by direct sequencing and CK2 phosphorylation sites were defined using a well-validated prediction rule. In vitro phosphorylation assays were performed using recombinant CK2 and synthetic peptides or full-length NS5A. In vivo phosphorylation by endogenous CK2 of NS5A expressed in hepatoma cells was also investigated. RESULTS: The mean number of CK2 sites within full-length NS5A, was significantly higher in HCV-3 compared to HCV-1a (P<0.01) and HCV-1b (P<0.01). The number of CK2 sites was more homogeneous in HCV-3 variants compared to HCV-1a and HCV-1b variants (P<0.05). The number of predicted CK2 sites correlated with the degree of in vitro and in vivo phosphorylation of NS5A by CK2. CONCLUSIONS: CK2-dependent phosphorylation of NS5A is heterogeneous among different HCV genotypes and clinical isolates. This might have an influence on virus biology and pathogenicity. PMID- 17923168 TI - Intra-species variation in transient accumulation of leaf anthocyanins in Cistus creticus during winter: evidence that anthocyanins may compensate for an inherent photosynthetic and photoprotective inferiority of the red-leaf phenotype. AB - Leaf color in some individuals of Cistus creticus turns transiently to red during winter, while neighboring individuals occupying the same site remain green. We have examined whether anthocyanin accumulation can be associated with variations in photosynthetic and/or photoprotective characteristics between the two phenotypes, rendering the red phenotype more vulnerable to photoinhibition and, accordingly, needing additional protection in the form of anthocyanins. Towards this aim, maximum (pre-dawn) and effective (mid-day) PSII photochemical efficiencies, xanthophyll cycle pool sizes and leaf nitrogen contents were seasonably followed, encompassing both the green (spring, summer, autumn) and the red (winter) period of the year. Moreover, the distribution of the two phenotypes in exposed and shaded sites was assessed. The frequency of red individuals was considerably higher in fully exposed sites, pointing to a photoprotective function of leaf anthocyanins. Yet, the assumption was not corroborated by pre dawn PSII yield measurements, since both phenotypes displayed similar high values throughout the year and a similar drop during winter. However, the red phenotype was characterized by lower light-saturated PSII yields, xanthophyll cycle pool sizes and leaf nitrogen, during both the green and the red period of the year. Based on this correlative evidence, we suggest that winter redness in C. creticus may compensate for an inherent photosynthetic and photoprotective inferiority, possibly through a light screen and/or an antioxidant function of leaf anthocyanins. PMID- 17923169 TI - Two types of aspartic proteinases from buckwheat seed--gene structure and expression analysis. AB - Two types of aspartic proteinase (AP) genes have been isolated from the cDNA library of developing buckwheat seeds. Analysis of their sequences showed that one of these, FeAP9, resembled the structure and shared high homology with the so called typical plant APs characterized by the presence of a plant-specific insert (PSI), an element unique among APs. The other cDNA, FeAPL1, encoded an AP-like protein lacking that domain. Different expression profiles were observed for FeAP9 and FeAPL1. FeAPL1 mRNAs were restricted to the seeds only, whereas FeAP9 mRNAs were also present in the other plant tissues - leaves, roots, and flowers. Higher levels of FeAP9 were observed in senescent leaves compared with green leaves. The differential expression pattern of these two unique APs raises the interesting possibility that these proteinases have unique substrate specificity and may have different roles in plant development and other physiological processes. PMID- 17923170 TI - Number, characteristics, and classification of patients with dermatomyositis seen by dermatology and rheumatology departments at a large tertiary medical center. AB - BACKGROUND: The current diagnostic criteria for dermatomyositis (DM) exclude patients without muscle involvement. As a result there is a paucity of research related to the complete spectrum of the disease. OBJECTIVE: The goal of this study was to evaluate differences in the clinical manifestations of DM seen by dermatology relative to rheumatology. We hypothesized that patients with minimal (hypomyopathic) or no (amyopathic) muscle disease would more likely be seen in dermatology, whereas those with more severe (classic) muscle disease would be seen in rheumatology. METHODS: We performed a retrospective chart review of patients with DM seen by our dermatology and rheumatology departments to classify spectrum, presentation, and complications. Patients seen between July 1, 2003, and June 30, 2006, were identified by Current Procedural Terminology billing code 710.3. Patients with mixed connective tissue diseases or miscoded DM were excluded. RESULTS: In all, 131 (65%) patients seen in dermatology, 58 (29%) in rheumatology, and 13 (6%) in both departments were identified. In all, 83 (69%) patients seen in dermatology, 27 (23%) in rheumatology, and 10 (8%) in both departments met criteria for inclusion in the study. The number of patients seen in rheumatology given the classification of classic DM (CDM) (24 of 27 [89%]), hypomyopathic DM (2 of 27 [7%]), and amyopathic DM (ADM) (1 of 27 [4%]) differed significantly from dermatology, where CDM comprised 27 of 83 (33%), hypomyopathic DM comprised 23 of 83 (28%), and ADM comprised 33 of 83 (40%) of the population, respectively (P < .001). Sex, ethnicity, and rates of interstitial lung disease differed between departments. There was no difference in the rates of interstitial lung disease between CDM and ADM (P = .30). The degree of muscle involvement did not correlate with the rates of DM-associated malignancy (P = .57). Few patients with ADM had muscle biopsy (n = 1) or electromyography (n = 7) testing. Positive anti-Jo-1 was seen in 2 of 96 patients (2%; one CDM and one ADM, both with interstitial lung disease), reflecting an overall low prevalence of this autoantibody, or a potential problem with the laboratory assay. LIMITATIONS: Patients reflect the population in only one institution and, thus, the results may not be generalizable to other settings or referral centers. Because this is a retrospective chart review, results are limited by missing data and nonstandardized physical examinations and laboratory data across patients and physicians. CONCLUSIONS: There is a clear difference in DM presentation to dermatology and rheumatology by degree of myositis-complicated disease. PMID- 17923171 TI - Convenient access to 1,3,4-trisubstituted pyrazoles carrying 5-nitrothiophene moiety via 1,3-dipolar cycloaddition of sydnones with acetylenic ketones and their antimicrobial evaluation. AB - Novel 1-aryl-3-(5-nitro-2-thienyl)-4-aroyl-pyrazoles 7 have been synthesized by the 1,3-dipolar cycloaddition of 3-arylsydnones 3 with 1-aryl-3-(5-nitro-2 thienyl)-2-propyn-1-ones 6. The newly synthesized compounds were well characterized by elemental analysis, IR, (1)H NMR and mass spectral studies. They were also screened for their antibacterial and antifungal activities against a variety of microorganisms and the results of such studies have been discussed in this article. PMID- 17923172 TI - Synthesis and anti-mycobacterial activity of (E)-N'-(monosubstituted benzylidene)isonicotinohydrazide derivatives. AB - A series of 22 (E)-N'-(monosubstituted-benzylidene)isonicotinohydrazide derivatives have been synthesized and evaluated for their in vitro antibacterial activity against Mycobacterium tuberculosis H(37)Rv using Alamar Blue susceptibility test and the activity expressed as the minimum inhibitory concentration (MIC) in mug/mL. Compounds 2f, 2g, 2j, 2k and 2q exhibited a significant activity (0.31-0.62 microg/mL) when compared with first line drugs such as isoniazid (INH) and rifampicin (RIP) and could be a good starting point to develop new lead compounds in the fight against multi-drug resistant tuberculosis. PMID- 17923173 TI - Vaccination against human papillomavirus: a baseline survey of Canadian clinicians' knowledge, attitudes and beliefs. AB - We assessed Canadian obstetrician/gynaecologists', family physicians' and paediatricians' knowledge, attitudes, and beliefs about HPV infection and prevention, as well as factors associated with willingness to prescribe HPV vaccines. A self-administered, anonymous questionnaire was mailed to 2500 physicians. The participation rate was 51%. Overall, 95% of respondents indicated that the vaccine should be given to girls before the onset of sexual activity; 80% of respondents felt that the best age for an HPV vaccination program is <14 years; 88% of respondents intend to recommend HPV vaccines if they are publicly funded. Several predisposing, reinforcing, organizational, preventive activity and situational factors were independently associated with the intention to prescribe the HPV vaccines. Most physicians expect an important benefit from HPV vaccination and intend to prescribe HPV vaccines (86.7%). To achieve high rates of vaccination coverage, several modifiable factors should be taken in consideration during immunization program implementation. PMID- 17923174 TI - Large-scale manufacture and use of recombinant VP2 vaccine against infectious bursal disease in chickens. AB - A new subunit vaccine against infectious bursal disease (IBD) was developed; the antigen used in the vaccine was expressed by a new engineering strain, E. coli BL21/pBV220-VP2. The study on the production and use of the vaccine was performed. The results showed that the recombinant VP2 was water-soluble and demonstrated natural antigen activity in vitro. The antibody induced by rVP2 subunit vaccine could protect chickens from challenges of IBDV strains, both BC 6/85 and JZ 3/02. The vaccine, in which the VP2 AGP titre is 1:4, would be enough to protect SPF chickens of 19-day-old, but seemed to be relatively lower to protect commercial Avian Broilers under 10-day-age. In field study, Avian Broilers were vaccinated with rVP2 subunit vaccine of 1:16 AGP titre at the age of 7 days. The protection rate was varied from 72% to 95% in different chicken farms. To study the method of serological evaluation, antibody respond to vaccination had been detected with three kinds of tests. The correct ratio of detection decreased in the order of VP2-based ELISA, AGP test, and virions-based ELISA, if the result of IBDV detection was used as standard of judging. Correlation coefficient between the OD values of VP2-based ELISA and the virions based ELISA was 0.782. The results will make it possible for the vaccine to be produced commercially and used in poultry industry in large scale. PMID- 17923175 TI - Digital photographs of excised lesions: an aid to histopathology reports. PMID- 17923176 TI - Numerical simulation of tooth movement in a therapy period. AB - BACKGROUND: Orthodontic tooth movements are based on the ability of bone reaction to mechanical stimulus with the deposition and resorption of alveolar bone. The numerical simulation of tooth movement could be helpful for the treatment strategy. However, at present, few calculations have been carried out on the tooth movement simulation. METHODS: Finite element (FE) models were developed to simulate an orthodontic treatment of mandibular canine tipping movement during a therapy period with decayed loads. The tooth movement was based on the surface bone remodeling method, and the normal strain of periodontal ligament was assumed as the key mechanical stimulus for alveolar bone remodeling. Changes in the tooth position and the geometry of the tooth supporting structures were taken into account. FINDINGS: The highest normal strain in the periodontal ligament was observed at the cervix or apex and the lowest normal strain was observed near the middle of the root. The tipping degrees of the simulation were similar to the observed in clinical studies. INTERPRETATION: It was acceptable to simulate clinical tooth tipping movements by finite element method based on these mechanical assumptions. Such a numerical simulation would be used to predict clinical tooth movements and help the planning of the therapy. PMID- 17923178 TI - Evaluation of stream ecological integrity using litter decomposition and benthic invertebrates. AB - Biomonitoring programs to access the ecological integrity of freshwaters tend to rely exclusively on structural parameters. Here we evaluated stream ecological integrity using (a) benthic macroinvertebrate derived metrics and a biotic index as measures of structural integrity and (b) oak litter decomposition and associated fungal sporulation rates as measures of functional integrity. The study was done at four sites (S1, S2, S3 and S4) along a downstream increasing phosphorus and habitat degradation gradient in a small stream. The biotic index, invertebrate metrics, invertebrate and fungal communities' structure and sporulation rates discriminated upstream and downstream sites. Decomposition rates classified sites S4 and S2 as having a compromised ecosystem functioning. Although both functional and structural approaches gave the same results for the most impacted site (S4), they were complementary for moderately impacted sites (S2 and S3), and we therefore support the need for incorporating functional measures in evaluations of stream ecological integrity. PMID- 17923177 TI - Flow shear stress affects macromolecular accumulation through modulation of internal elastic lamina fenestrae. AB - BACKGROUND: Internal elastic lamella and elastic lamellae serve as barriers for macromolecular accumulation in vascular wall. The function of internal elastic lamella in atherogenesis induced by shear stress has not been well understood yet. In the present study, internal elastic lamella remodeling and transmural accumulation of macromolecules in the arterial wall under altered shear stress were investigated. METHODS: The ligation of left distal carotid branches induced low shear stress in the left common carotid artery and high shear stress in the right common carotid artery of rats for 7 days. Fenestrae in internal elastic lamella were assessed by confocal microscopy. Horseradish peroxidase accumulation was determined by its transmural concentration profiles. Fibronectin expression was examined by immunohistologic staining and Western blotting. FINDINGS: Low shear stress significantly decreased the percentage of fenestrae area in internal elastic lamella from 15.35% to 8.03% in left common carotid artery, while high shear stress nearly had no influence in right common carotid artery. Comparing with less accumulation of horseradish peroxidase in right common carotid artery, horseradish peroxidase was significantly accumulated in the innermost part of the media, and fibronectin expression increased to 3.3-fold in left common carotid artery. INTERPRETATION: Our results suggested that shear stress alteration could influence the endothelial cell function as well as internal elastic lamella and elastic lamellae remodeling. Low shear stress decreased the permeability of internal elastic lamella and elastic lamellae by reducing fenestrae of them, which involved the enhanced fibronectin expression. Therefore, the increased accumulation of macromolecules in the arterial wall was observed. Our data supply new evidence on the mechanisms of atherogenesis induced by low shear stress. PMID- 17923179 TI - Lay accounts of depression amongst Anglo-Australian residents and East African refugees. AB - Layperson accounts of depression are gaining increasing prominence in the health research literature. This paper considers the accounts of lay people from a cross cultural perspective. By exploring lay concepts of distress from Anglo Australian, Ethiopian and Somali communities in Australia, we describe commonalities and divergences in understandings of depression. A total of 62 Anglo-Australians were interviewed, and 30 Somali and Ethiopians participated in focus groups and individual interviews. Anglo-Australian accounts frequently portray depression as an individual experience framed within narratives of personal misfortune, and which is socially isolating. In the accounts of distress from the Somali and Ethiopian refugees living in Australia, family and broader socio-political events and circumstances featured more frequently, and 'depression' was often framed as an affliction that was collectively derived and experienced. PMID- 17923180 TI - Acculturation and the use of complementary and alternative medicine. AB - The use of complementary and alternative medicine (CAM) has been growing substantially in the US in recent years. Such a growth in CAM use coincides with an ongoing increase in the proportion of the foreign-born population in the US. The main objective of this study is to examine the relation between acculturation and the use of CAM therapies among immigrants. Data from a CAM supplement to the 2002 National Health Interview Survey were analyzed to estimate the effects of acculturation on the likelihood of using different CAM therapies over the past 12 months prior to the survey. The results suggest that the level of acculturation as measured by nativity/length of stay in the US and language of interview-is strongly associated with CAM use. As immigrants stay longer in the US or as their use of English becomes more proficient, the likelihood that they use CAM therapies increases as well, and it gradually approaches the level of CAM use by native-born Americans. Moreover, this relation between acculturation and CAM use generally persists even after the effects of socioeconomic status, health insurance coverage, self-reported health status, and other demographic variables have all been taken into consideration. The substantially lower rates of CAM use by recent immigrants and its possible causes warrant further research. PMID- 17923182 TI - Congenital pouch colon associated with anorectal malformation: report of 2 cases. AB - Congenital pouch colon associated with anorectal malformation is an unusual anomaly reported most frequently in Asian countries. Pediatric surgeons must be familiar with this anomaly and the pre- and postoperative problems of these patients. The aim of this report is to bring attention to congenital pouch colon associated with anorectal malformation by discussing experiences with 2 patients. PMID- 17923181 TI - Colonic adenocarcinoma in a 13-year-old with cystic fibrosis. AB - The increase in longevity among patients with cystic fibrosis has brought to light comorbidities associated with the disease that are less acutely fatal than its pulmonary manifestations. Modern retrospective analysis has demonstrated an increased risk of gastrointestinal malignancy in patients with cystic fibrosis, with a marked elevation in malignancy rates among patients in their 20s and 30s. Here we report a case of colonic adenocarcinoma presenting as pneumaturia in a 13 year-old patient with cystic fibrosis. PMID- 17923183 TI - Cholelithiasis and choledocholithiasis associated with anomalous junction of the cystic duct in a child. AB - We report an extremely rare case of cholelithiasis, presumably owing to cholestasis resulting from an anomalous course of the cystic duct. A 10-year-old girl visited our hospital because of right epigastric pain and fever. Cholelithiasis and choledocholithiasis were diagnosed by ultrasound examination. Magnetic resonance cholangiopancreatography showed no pancreaticobiliary maljunction but confirmed a dilated, tortuous cystic duct anomalously draining into the right hepatic duct. Because cholangitis and obstructive jaundice progressed after admission, emergent endoscopic retrograde cholangiopancreatography was performed, and a common bile duct stone was removed endoscopically. It was a bilirubin stone. At a later date, laparoscopic cholecystectomy was performed for cholelithiasis. Preoperative 3-dimensional computed tomography and intraoperative cholangiography enabled us to treat the cystic duct safely. PMID- 17923184 TI - Intestinal chondrolipoma: uncommon cause of bowel obstruction. AB - A case of intestinal obstruction caused by jejunal chondrolipoma in an 11-year old boy is presented. To the best of our knowledge, this seems to be the first report of chondrolipoma arising from the small bowel in English language literatures. PMID- 17923186 TI - Neurogenic bladder decompressing via a spontaneous colovesical fistula--a case report. AB - A 3-year-old girl with operated meningomyelocele and urinary incontinence presented with recurrent attacks of watery diarrhea and anuria, which were relieved by urethral catheterization. Investigations revealed a poorly compliant neurogenic bladder that periodically decompressed via a spontaneously developed colovesical fistula. The fistula was repaired with concomitant bladder augmentation. To our knowledge, this is the only report of such a complication from neurogenic bladder in childhood. PMID- 17923185 TI - Immature teratoma of gallbladder associated with gliomatosis peritonei, a case report. AB - The patient was a 9-month-old girl who presented with an abdominal mass. Computed tomography scan revealed that the huge heterogeneous mass with cystic, calcified, and solid components occupied the upper abdomen, and diagnosis of teratoma was made preoperatively. At the operation, a tumor arising from gallbladder and gliomatosis peritonei with intact ovaries were found. This is a rare case report of teratoma that originated in the gallbladder associated with gliomatosis peritonei. PMID- 17923187 TI - Superior mesenteric artery syndrome in an infant: case report and literature review. AB - Superior mesenteric artery syndrome (SMAS) is an obstruction at the third portion of the duodenum by compression between the superior mesenteric artery and the aorta. In infancy, SMAS is extremely rare; and for its diagnosis, other duodenal obstructive diseases including congenital duodenal stenosis and intestinal malrotation must be ruled out. We present the case of a 7-month-old girl with frequent bilious vomiting after the resolution of acute gastroenteritis. Superior mesenteric artery syndrome was finally diagnosed at laparotomy, and duodenojejunostomy was performed. Vomiting disappeared postoperatively, and she gained weight. Although SMAS is an extremely rare syndrome in infants, it should be considered as a possible cause of incomplete duodenal obstruction. PMID- 17923189 TI - Predictors of gastroesophageal reflux in neonates with congenital diaphragmatic hernia. AB - PURPOSE: Gastroesophageal reflux (GER) is observed in 22% to 81% of neonates with congenital diaphragmatic hernia (CDH). The purpose of this study was to identify factors that may predict GER requiring fundoplication in neonates with CDH. METHODS: A retrospective chart review was performed on all neonates with CDH treated at our hospital from June 1997 to June 2005. Preoperative respiratory status, side of the CDH, and method of repair were assessed as predictors of GER and the need for fundoplication. RESULTS: Of the 42 patients with CDH, 3 died before intervention, leaving 39 patients eligible for study. All but 1 patient survived until discharge. Twenty-one (54%) developed GER of whom 9 (23%) required fundoplication. Although the side of the CDH was not a determinant of GER or the need for fundoplication, patch repair and the need for extracorporeal life support were determinants of both. CONCLUSIONS: Gastroesophageal reflux is common among babies with CDH, although symptoms often resolve without surgical intervention. Infants with CDH defects requiring a patch repair and those requiring advanced physiologic support, especially extracorporeal life support, are likely to develop severe GER necessitating fundoplication. Early recognition and treatment of GER among high-risk patients may shorten hospital stay and minimize patient morbidity. Early fundoplication should be considered for those patients at the highest risk. PMID- 17923188 TI - Continuous remission in an infant with chest wall malignant rhabdoid tumor after relapse. AB - Malignant rhabdoid tumor (MRT) is a highly aggressive tumor that occurs in infancy or childhood. The prognosis, especially in infants, is very poor. Here we report the long-term survival of a 5-month-old boy with MRT that arose from the chest wall. After total resection of the tumor, the patient was given 4 cycles of doxorubicin, vincristine, and cyclophosphamide, alternating with ifosfamide and etoposide. After 18 months off therapy, he had a local recurrence at the same site. After a second total resection, he was given additional chemotherapy with 30.6-Gy local irradiation. No further recurrence has been observed for 5 years since the second complete remission. Currently, he is alive and well at 7.5 years post-onset. Our experience in this case suggests a fundamental strategy of successful treatment of this highly malignant pediatric tumor: (1) complete resection of the localized tumor, (2) intensive multiagent chemotherapy for the minimal disseminated disease, and (3) radiotherapy for local control of the disease. PMID- 17923191 TI - Adriamycin produces a reproducible teratogenic model of vertebral, anal, cardiovascular, tracheal, esophageal, renal, and limb anomalies in the mouse. AB - BACKGROUND/PURPOSE: The Adriamycin rat model is an established model for vertebral, anal, cardiac, tracheal, esophageal, renal, and limb (VACTERL) anomalies and gastrointestinal atresias. Mice are the foremost mammal studied by developmental biologists, providing greater availability of molecular probes, antibodies, and transferable knowledge with transgenic studies. Only tracheoesophageal malformations have been previously described in the Adriamycin mouse model. The aim of this study was to carry out a dose-response analysis of the teratogenicity of Adriamycin in the mouse to determine the effect of the dose and timing of exposure in producing tracheoesophageal malformations and show if it causes other VACTERL anomalies. METHODS: CBA/Ca mice were accurately time mated (n = 30). Four different doses (0 [saline], 4, 5, and 6 mg/kg) of Adriamycin (EBEWE Pharma Ges.m.b.H. Nfg.KG, A-4866 Unterach, Austria) at 3 different timings of injections were compared. Dams received 2 intraperitoneal injections, 24 hours apart, commencing on day 7, 7.5, or 8. Fetuses were harvested on day 18. Anomalies were examined using a dissecting microscope and serial transverse sections. RESULTS: Administering Adriamycin at 6 mg/kg on days 7 and 8 had the most teratogenic effect, with 80% of fetuses having 3 or more VACTERL anomalies: anorectal malformation, 100%; tracheoesophageal malformation, 50%; right-sided aortic arch, 58.3%; bladder agenesis/bilateral hydronephrosis, 100%. CONCLUSION: This study establishes a mouse model that should provide insights into the cellular and molecular mechanisms underlying VACTERL anomalies. PMID- 17923190 TI - Effect of antenatal tetrandrine administration on endothelin-1 and epidermal growth factor levels in the lungs of rats with experimental diaphragmatic hernia. AB - PURPOSE: The aim of this study was to evaluate the effect of the traditional Chinese medicine tetrandrine (Tet) and to determine its possible mechanism on expression of endothelin-1 (ET-1) and epidermal growth factor (EGF) in the lung of a rat model of nitrofen-induced congenital diaphragmatic hernia (CDH). METHODS: A single oral dose (115 mg/kg) of nitrofen on day 9.5 of pregnancy was maternally administered to induce CDH. Pregnant rats were divided into 4 groups on day 18.5: control (n = 5), CDH (n = 5), CDH+dexamethasone (Dex) (n = 5), and CDH+Tet (n = 5). All fetuses were delivered by cesarean delivery on day 21.5. Accordingly, there were 4 groups of fetuses: control (n = 38), CDH (n = 25), CDH+Dex (n = 21), and CDH+Tet (n = 22). Lung tissue weight (LW) and body weight (BW) of each fetus were recorded, lung histologic evaluations and ET-1 and EGF immunohistochemistry staining were performed, and image analysis was performed after lung processing. RESULTS: Five female rats in the control group produced 38 fetuses without CDH. CDH was observed in 68 of the 128 rat fetuses (53.1%) among the other 3 groups. The LW/BW ratio of the CDH group was significantly lower than those of the Dex and EGF groups (P < .05). The lungs of fetuses with CDH showed marked abnormal structure such as pulmonary hypoplasia and vascular remodeling, in contrast to improved pulmonary structure in lungs of fetuses in the CDH+Dex and CDH+Tet groups. Statistical differences in morphologic parameters (radial alveolar counts, percentage of alveoli, percentage of medial wall thickness, and vascular volume) were found (P < .05). The immunoreactivity of EGF and ET-1 in the CDH group was markedly stronger than that in the control, CDH+Dex, and CDH+Tet groups (P < .01). In addition, EGF and ET-1 expression in the CDH+Dex and CDH+Tet groups was stronger than that in the control group (P < .05). There was no difference in lung EGF and ET-1 immunoreactivity between CDH+Dex and CDH+Tet groups (P > .05). CONCLUSION: Antenatal treatment with Tet may improve lung growth and vascular remodeling, and its mechanism seems to be involved in decreasing EGF and ET-1 expression. Tet administered maternally may be a hopeful new therapeutic option in the treatment of CDH and may be effective in helping to avoid the side effects of Dex. PMID- 17923192 TI - Long gap esophageal atresia: lengthening technique and primary anastomosis. AB - OBJECTIVE: The treatment of long gap esophageal atresia remains a major surgical challenge. The authors describe a modification of a lengthening technique based on tissue expansion to avoid sutures cutting through the esophagus. METHODS: Between January 2004 and August 2006, 4 patients did not respond to stretching, and underwent this modified esophageal lengthening technique using silastic tubes. RESULTS AND FOLLOW-UP: All infants recovered and have an intact esophagus. All infants developed gastroesophageal reflux. Thal antireflux procedure was performed in the first infant. The other 3 patients were managed conservatively. Follow-up ranged between 6 and 34 months. CONCLUSIONS: The tissue expansion principle can be successfully applied in the esophagus through external traction. Silastic tube fixation at esophageal ends may help to apply even traction and avoid sutures cutting through the esophageal tissue. PMID- 17923193 TI - Intracellular calcium mobilization of the aganglionic intestine in the endothelin B receptor gene-deficient rat. AB - BACKGROUND AND AIM: Up to now, numerous reports have analyzed the pathogenesis of Hirschsprung's disease (HD) by means of physiologic, pathologic, or molecular biologic methods. However, very little is still known about the smooth muscle cell itself. The endothelin B receptor gene-deficient (EDNRB(-/-)) rat, which is suitable for research of HD, has an aganglionic segment of the total colon. Our purpose is to investigate the myogenic mechanisms using simultaneous measurements of the intracellular Ca2+ concentration ([Ca2+]i) and tension and reverse transcriptase polymerase chain reaction for L-type Ca2+ channel (L-VOC) expression. METHODS: The muscle strips of the rat distal colon were loaded with a Ca2+ indicator dye, fura-PE3/AM, for 3 to 4 hours. The changes in the fluorescence intensity of Ca2+-fura-PE3 complex of the strips were monitored with a front surface fluorometer (CAM-230). The fluorescence intensities at 340- and 380-nm excitation and their ratio (F340/F380) were recorded as the level of [Ca2+]i. The comparison of L-VOC alpha1c subunit messenger RNA (mRNA) expression in both wild and homozygous rat was performed by reverse transcriptase polymerase chain reaction. RESULTS: The peak levels of force development induced by carbachol were 139.1% +/- 5.0% in EDNRB(-/-) rat, whereas the peak levels were 242.1% +/- 27.7% in EDNRB(+/+) rat. The changes in the [Ca2+]i elevation induced by carbachol were 101.7% +/- 12.2% in the homozygous rat, whereas these were 143.8% +/- 8.9% in the wild-type rat. Both results in the homozygous rat significantly decreased in comparison with those of the wild rat (P < .05). The expression of the L-VOC channel mRNA also decreased in the homozygous rat. CONCLUSIONS: This is the first report to show the [Ca2+]i mobilization in the smooth muscles of the rat model of HD. The decrease in both [Ca2+]i and force development was thus considered to be due to the decrease in the Ca2+ channel expression. PMID- 17923194 TI - Ten-year experience in the management of total colonic aganglionosis. AB - PURPOSE: The aim of this study was to review the 10 years' experience in the management of patients with total colonic aganglionosis (TCA) and follow-up of their health condition. METHODS: Cases of 25 patients with TCA in the Children's Hospital of Fudan University from 1996 to 2005 were reviewed and analyzed. The confirmed diagnosis was established by an intraoperative frozen-section biopsy of the rectum, colon, appendix, and ileum. The data included in this study accounted for sex, age, signs of presentation, any familiar history of Hirschsprung disease (HD) or associated abnormalities, and ileal involvement. Plain x-ray films, barium enema, and anorectal manometry were provided for evaluation. The results of surgical management were analyzed for weight at definite operation, blood requirement during operation, the total parenteral nutrition duration, and the pre- and postoperative complications of these patients. Follow-up data were collected regarding growth development, stool frequency, stool consistency, fecal soiling, incontinence, enterocolitis, and anal stricture. RESULTS: Among 25 patients, 8 (32%) females and 17 (68%) males were diagnosed as having TCA. Sixteen patients (64%) were evaluated at the neonatal period, whereas 9 patients (36%) were evaluated after the neonatal period. All 25 patients received at least 1 plain abdominal radiograph or barium enema at the university hospital before operation. However, there was no specific pathognomonic finding that may provide a definite diagnosis. Nineteen (76%) patients underwent initial laparotomy at our institute and 6 patients (24%) were operated on beforehand at other hospitals. Twenty-three (92%) patients were diagnosed as having TCA and underwent ileostomy, whereas 2 (8%) patients underwent primary pull-through procedure. Eighteen (72%) patients had undergone definite surgery. Pre- and postoperative complications included enterocolitis (44.4%), perianal excoriation (77.7%), electrolyte imbalance (50%), and anastomotic leak (16.6%). Average duration of total parenteral nutrition before operation was 17.77 +/- 12.54 days and after operation was 10.27 +/- 5.23 days. Mean follow-up time was 27.6 +/- 35.39 months. Two patients had 5 to 6 bowel movements per day. Seven had a frequency of stool ranging between 1 and 3 bowel movements per day. Their bowel movements returned to normal about 12 to 18 months after surgery. On follow-up, the height and weight development of the patients was found to be normal. CONCLUSIONS: Gradual progress was observed in all the patients that took part in the study, and all patients had positive results eventually. PMID- 17923195 TI - Calcium carbonate gallstones in children. AB - BACKGROUND: In the United States, cholesterol stones account for 70% to 95% of adult gallstones and black pigment stones for most of the remainder. Calcium carbonate stones are exceptionally rare. A previous analysis of a small number of pediatric gallstones from the north of England showed a remarkably high prevalence of calcium carbonate stones. The aims of this study were to analyze a much larger series of pediatric gallstones from our region and to compare their chemical composition with a series of adult gallstones from the same geographic area. METHODS: A consecutive series of gallbladder stones from 63 children and 50 adults from the north of England were analyzed in detail using Fourier transform infrared microspectroscopy. Demographic and clinical data were collected on all patients. The relative proportions of each major stone component were assessed: cholesterol, protein and calcium salts of bilirubin, fatty acids, calcium carbonate, and hydroxyapatite. RESULTS: Thirty-nine (78%) adults had typical cholesterol stones, 7 (14%) had black pigment bilirubinate stones, and only 2 (4%) had calcium carbonate stones. In contrast, 30 (48%) children had black pigment stones, 13 (21%) had cholesterol stones, 15 (24%) had calcium carbonate stones, 3 (5%) had protein dominant stones, and 2 (3%) had brown pigment stones. In children, cholesterol stones were more likely in overweight adolescent girls with a family history of gallstones, whereas black pigment stones were equally common in boys and girls and associated with hemolysis, parenteral nutrition, and neonatal abdominal surgery. Calcium carbonate stones were more common in boys, and almost half had undergone neonatal abdominal surgery and/or required neonatal intensive care. CONCLUSION: The composition of pediatric gallstones differs significantly from that found in adults. In particular, one quarter of the children in this series had calcium carbonate stones, previously considered rare. Geographic differences are not the major reason for the high prevalence of calcium carbonate gallstones in children. PMID- 17923196 TI - Laparoscopic treatment of hydatid cyst of the liver in children. A report on 34 cases. AB - PURPOSE: This study evaluated the safety and efficiency of laparoscopically treated liver cysts in children. METHODS: From September 2001 to July 2004, 34 patients underwent laparoscopic treatment of hydatid cysts of the liver. All patients had chest x-ray, abdominal sonography, and hydatid serology. The different stages of the procedure were the same as in open surgery: puncture, aspiration, injection of scolicidal agent, reaspiration, removal of proligerous membrane, and resection of the dome. RESULTS: The patients' mean average age was 7 years and 7 months (range, 3-14 years). The number of cysts ranged from 1 to 10 with a diameter of 40 to 150 mm (mean diameter, 65.5 mm). One case had a mesenteric associated hydatid cyst, another splenic hydatid cyst. The average length of hospital stay was 5 days (range, 4-14 days). No per- or postoperative complications were reported. At 12 to 45 months follow-up, no recurrence has been reported. CONCLUSION: Laparoscopy represents an excellent approach for the treatment of hydatid cyst of the liver in children. PMID- 17923197 TI - Enteral resveratrol supplementation attenuates intestinal epithelial inducible nitric oxide synthase activity and mucosal damage in experimental necrotizing enterocolitis. AB - BACKGROUND/PURPOSE: The release of various enzymes including inducible nitric oxide synthase (iNOS) leads to enterocyte apoptosis through free nitrogen radicals, which in turn leads to impaired mucosal barrier and bacterial translocation with resultant sepsis in necrotizing enterocolitis (NEC). Resveratrol, a polyphenol compound from phytoalexins with antioxidant and scavenger properties, also play a critical role in modulating key enzymes in cell cycle including iNOS. We therefore hypothesized that resveratrol would prevent mucosal damage in experimental NEC in rats. METHODS: Newborn rats were randomized into 3 groups: group 1 was left to breast-feed (BF), whereas group 2 (NEC) was induced by enteral formula feedings twice daily and by being subjected to hypoxia thrice. The third group (R) received the same treatment as the NEC group but the enteral feeds were supplemented with resveratrol. Rats were killed on day 4, and their terminal ileal samples were harvested for histopathologic analysis. Expression of iNOS was assessed by sodium dodecyl sulfate polyacrylamide-gel electrophoresis analysis and immunohistochemistry. Band densities were quantified by using the software NIH image. RESULTS: The epithelial structure in group BF was normal. In the NEC group, there were marked loss of the brush border, vacuolization, and necrosis. The epithelial structure was found to be preserved in group R. Western blot analysis revealed marked elevation in the expression of iNOS protein at 130 kD molecular weight (band densities in groups BF, NEC, and R were 0.3 +/- 3.5, 3.7 +/- 2.9, and 0.6 +/- 5.1, respectively; P < .01). Immunohistochemical analysis revealed that iNOS staining was significantly increased in the NEC group, whereas it remained minimal for the BF and R groups. Ileal tissue nitrate/nitrite levels for groups BF, NEC, and R were 178.3 +/- 7, 191.4 +/- 4.1, and 181 +/- 3.6 micromol/(L x g), respectively (P < .01). CONCLUSIONS: These findings may provide insights for the beneficial effect of enteral resveratrol supplementation on inflammatory conditions of the bowel including NEC through attenuating the release of iNOS and preservation of mucosal integrity. PMID- 17923198 TI - Angiographic, ultrasonographic, and macroscopic assessment of aortic growth after VCS clips, interrupted polypropylene, or running polyglycolic acid anastomosis. AB - BACKGROUND/PURPOSE: Lack of growth after surgery is still an unsolved issue in growing vessels. Aortic surgery is an integral part of vascular pediatric surgery. As an alternative to address this lack of growth in this scenario, we propose the use of titanium clips for vascular anastomosis. METHODS: Thirty-two domestic swine were used in this study. Animals entered the study when they were 55 days old and were subjected to an end-to-end aortic anastomosis using vascular closure stapler (VCS) clips, interrupted polypropylene, or running polyglycolic acid suture. Control group animals were sham-operated. Pigs were allowed to grow for 6 months, during which time they were subjected to serial ultrasonographic and angiographic studies to assess vascular growth. RESULTS: VCS clip anastomosis outcome was comparable with polypropylene or polyglycolic acid suture anastomosis. No significant differences in either longitudinal or transversal vascular growth were seen between the 3 studied suturing techniques and the control group. CONCLUSIONS: Our vascular growth and patency results suggest that VCS clips may provide a suitable alternative to conventional suture in pediatric aortic vascular surgery. PMID- 17923199 TI - Omphalocele induction in the chick embryo by administration of cadmium. AB - BACKGROUND: Ventral body wall (VBW) defects occur in 1:2000 live births. We examined the association of VBW defect with somite abnormality and lordosis in the chick using in vitro and in ovo methods. METHODS: Explanted chick embryos were treated at 60 hours with 50 microL sodium acetate or 0.001% cadmium acetate solution to produce VBW defects. Mortality and abnormality rates were assessed. A further cohort of chicks was treated in ovo by dropping 50 microL 0.001% to 0.01% cadmium acetate onto the embryo and allowing development to 16.5 days for further assessment of the defect and skeletal staining with alcian blue and alizarin red. RESULTS: Cadmium treatment at 24 hours induced VBW defects in chicks treated in both shell-less culture and in ovo. Material herniating through the VBW defects was covered by a membrane in all fresh specimens. Membrane removal revealed large defects containing liver and bowel. These criteria clearly indicate that the defect observed is an omphalocele. Affected embryos had reduced somite numbers within 24 hours. Chicks exhibiting exomphalos at 16.5 days invariably had lumbosacral lordosis. CONCLUSIONS: The cadmium-treated chick embryo is a reliable model for exomphalos. A positive association was found between exomphalos and lumbar lordosis in the chick. PMID- 17923200 TI - Amniotic fluid ferritin as a marker of intestinal damage in gastroschisis: a time course experimental study. AB - BACKGROUND/PURPOSE: Intestinal damage (ID) is closely related to morbidity and mortality in gastroschisis. This study was performed to determine the intraamniotic substances that may correlate ID and also to verify their time course levels that would be useful for determining when ID starts in gastroschisis. METHODS: In this study, 13-day-old fertilized chick eggs were used. The amnioallantoic membrane was perforated to create amnioallantoic cavity in all embryos. Gastroschisis was created in gastroschisis group to simulate human gastroschisis. Amnioallantoic fluid samples were collected from the embryos on the 13th to 19th gestational days, and the intestines of each group were harvested for evaluation. Amnioallantoic levels of interleukin-8, ferritin, alkaline phosphatase, and amylase were measured. Serosal thickness of the intestines in each group was evaluated. RESULTS: Increasing amnioallantoic fluid levels of interleukin-8, alkaline phosphatase, and amylase were found in both groups. In contrast to control group, ferritin levels, as a sign of inflammation, were found increased only in gastroschisis group. Histopathologic examination of intestines in the gastroschisis group showed a significant increase in the serosal thickness especially after the 16th day. CONCLUSION: Increases in amnioallantoic fluid levels of ferritin show promise as a marker for determining ID encountered in gastroschisis but warrant further investigation. PMID- 17923201 TI - Assessment of value of the sentinel lymph node biopsy in melanoma in children and adolescents and applicability of subcutaneous infusion anesthesia. AB - BACKGROUND/PURPOSE: The management of pediatric melanoma is controversial but equates that of adults. Lymphatic mapping with sentinel lymph node (SLN) biopsy is proposed as standard of care for patients with primary melanoma. The operation can be done with general or local anesthesia in adults. The goal of this study was to determine the applicability of subcutaneous infusion anesthesia (SIA) for SLN biopsy in children and adolescents, as well as to assess complications of this procedure and to document outcome of patients with melanoma in this particular age group after SLN biopsy. METHODS: Charts of patients with melanomas on the trunk and extremities who underwent lymphatic mapping and SLN biopsy in SIA between November 2000 and January 2006 revealed 13 patients with age 21 years or less. Tumescent solutions with lidocaine (0.2%) were used for SLN biopsy. Patient demographics, tumor thickness, Clark level, location of primary melanoma, ulceration, number of SLNs, number of positive nodes, and follow-up of patients were included. RESULTS: In 13 patients (age range, 12-21), SLN biopsy was performed. Mean tumor thickness was 1.8 mm (range, 1.0-7.0), none of these melanomas showed ulceration. The operation was tolerated in SIA by all patients; none had any associated complications. Of 13 patients, 5 (38.5%) had positive sentinel nodes. Three patients underwent completion lymph node dissection, and no further positive nodes were found. After a mean follow-up of 29.2 months (range, 13-68), all patients were found disease-free. CONCLUSIONS: Sentinel lymph node biopsy in SIA can safely be performed in children and adolescents with primary melanomas. Further studies are necessary to determine the prognostic information and therapeutic implications of SLN biopsy in this patient group. PMID- 17923203 TI - Thoracoscopic resection of neurogenic tumors in children. AB - PURPOSE: The aim of this study was to evaluate the feasibility of thoracoscopy in neurogenic tumors in infants and children. MATERIALS AND METHODS: From January 2000 to October 2005, 21 patients aged 7 months to 14 years (mean, 6 years) underwent thoracoscopy for tumor resection in 5 French institutions. One 10-mm optical port and 2 operative 5-mm ports were needed. Selective intubation was required for 3 patients aged about 12 years. Tumor was removed with an endoscopic bag in all cases. RESULTS: All procedures were completed successfully without any incomplete resection or recurrence. One conversion was necessary because of a huge mass. A chest tube was left for a mean of 2 days for 17 children. Two children had not had any drainage. Two postoperative chylothorax required chest drainage for 12 days. Only 5 of the 6 older patients (mean age, 12 years) needed a patient-controlled analgesia. The mean operative time was about 100 minutes. Hospital stay ranged from 4 to 12 days. Tumors were neuroblastoma or ganglioneuroblastoma in 16 cases and ganglioneuroma in the 5 other cases. CONCLUSION: Thoracoscopy for resection of thoracic neurogenic tumors in children is a feasible, safe, and efficient procedure. The surgeon has a better visualization of the tumor and its anatomic connections. Resection can be as complete as an open procedure without having to complicate the operative technique in the same operating time. It avoids cosmetic and functional disorders because of thoracotomy. It allows a good cosmetic resection without spillage. PMID- 17923202 TI - Tumors of dysgenetic gonads in Swyer syndrome. AB - BACKGROUND/PURPOSE: The female with Swyer syndrome requires close follow-up because of the high risk of neoplastic transformation in the dysgenetic gonads. The aim of this work was to present our experience with tumors in patients with Swyer syndrome. METHODS: We studied 8 females with Swyer syndrome. At the time of diagnosis, they were 13 to 18 years old. We performed an ultrasound examination of dysgenetic gonads, hormonal (follicle-stimulating hormone, luteinizing hormone, estradiol, and testosterone) and genetic (SRY, karyotype) tests, and histologic analysis of gonads (bilateral gonadectomy was performed in all patients). RESULTS: Gonadal tumors were found in 6 patients (3 cases of gonadoblastoma, 1 dysgerminoma, and 2 gonadoblastoma with dysgerminoma). Hormonal activity of gonadoblastoma was noted in 3 patients, with 1 tumor producing androgens. CONCLUSION: Our data suggest that patents with gonadal dysgenesis and 46,XY karyotype should be referred for bilateral gonadectomy because of the high risk of neoplastic transformation. Estrogen-producing gonadoblastoma may mask gonadal dysgenesis and delay the diagnosis of this pathology. PMID- 17923204 TI - In vitro study of ingested coins: leave them or retrieve them? AB - BACKGROUND: Objects and notably coins are frequently swallowed by children 3 to 5 years old. Precisely how they should be managed after passing the gastroesophageal junction without causing symptoms remains controversial. This study was performed to assess dissolution of specific metals from coins immersed in simulated gastric juice. METHODS: Four types of euro and US coins were immersed in simulated gastric juice for 4, 24, 72, and 120 hours. Six metals were evaluated by inductively coupled plasma-atomic emission spectrometry. Weight loss and corrosive behavior were also determined. RESULTS: After only 4 hours, metals had dissolved from euro (Cu, 2.86-7.85 mg; Ni, 0.23-0.52 mg; Zn, 0.09-0.99 mg; Al, 0.24 mg; Sn, 0.02 mg) and US (Cu, 1.45-6.65 mg; Ni, 0-0.62 mg; Zn, 0-0.14 mg) coins. Their concentrations in simulated gastric juice peaked after 24 to 72 hours (milligrams/hours) in euro (Cu, 218/48; Ni, 82.50/72; Zn, 83.00/72; Al, 14.65/72; Sn, 0.66/72) and US (Cu, 126.50/24; Ni, 88.00/72; Zn, 149.00/24) coins. All coins underwent corrosion and weight loss (by 2.56%-4.8%). CONCLUSIONS: Coins retained in the stomach will release a number of heavy metals well known to cause dose-dependent poisoning. Studies to evaluate their toxicity and absorption are needed to optimize treatment. PMID- 17923205 TI - Temporal factors and the incidence of physical abuse in young children: decreased nonaccidental trauma during Child Abuse Prevention Month. AB - BACKGROUND/PURPOSE: Educational efforts have been shown to decrease the incidence of abusive injuries. Information related to factors that associate with child abuse would further these educational efforts. We undertook this study to determine the influence of temporal factors and a national preventative program on the incidence of abusive injuries in young children. METHODS: The Pennsylvania Trauma Outcome Study database was queried for all children 36 months old or younger admitted with abusive injuries. Data collected included day, month, and season. Data were analyzed by Poisson regression. RESULTS: Six hundred seventy two cases of child abuse were reported, with a linear increase in the number of cases per year (P < .001). Sunday had fewer abuse cases admitted than any other day of the week (risk ratio, 0.70; P = .008). Younger children were less likely to be injured during April, which is National Child Abuse Prevention Month, and more likely to be injured during August and October (all P < .05). CONCLUSIONS: We observed fewer inflicted injuries in young children on Sundays, and during April. The causes for these significant variations deserve further study. PMID- 17923206 TI - Dehydroepiandrosterone treatment attenuates reperfusion injury after testicular torsion and detorsion in rats. AB - PURPOSE: We aimed to evaluate the effects of dehydroepiandrosterone (DHEA) on antioxidant enzyme activities, lipid peroxidation, and histopathologic changes in both testes after unilateral testicular torsion and detorsion. METHODS: Twenty four adult male Sprague-Dawley rats were randomly divided into 4 groups (n = 6 for each group): sham operation, torsion/detorsion (T/D), T/D + vehicle, and T/D + DHEA. Three hours before detorsion, 50 mg/kg DHEA was given intraperitoneally to the T/D + DHEA group. Testicular ischemia was achieved by twisting the left testis 720 degrees clockwise for 3 hours, and reperfusion was allowed for 24 hours after detorsion. In all groups, bilateral orchiectomies to determine the testicular tissue catalase (CAT) and superoxide dismutase activities and malondialdehyde (MDA) levels and histopathologic examination were performed. RESULTS: Compared with those from the sham group, CAT activities in the ipsilateral testis obtained from the T/D group were significantly lower and MDA levels were significantly higher (P < .05 for all). Administration of DHEA prevented increases in MDA levels and decreases in CAT and superoxide dismutase activities when compared to the T/D group. Specimens from the T/D and the T/D + vehicle groups had a significantly greater histologic injury than the specimens from the sham and the T/D + DHEA groups had (P < .01 for both). CONCLUSIONS: The results suggest that DHEA may be a protective agent for preventing biochemical and histopathologic changes related to oxidative stress in testicular injury caused by testis torsion. PMID- 17923207 TI - Cryptorchidism: the veracity of online information accessible to the public. AB - PURPOSE: The objective of this study is to assess the quality of information regarding cryptorchidism using common Internet resources. METHODS: Five common search engines were queried with the string "cryptorchidism OR undescended testicle OR undescended testes OR retractile testes OR retractile testicle." The first 30 sites retrieved via each engine were reviewed, and the content veracity of the following components was assessed independently by 2 pediatric urologists: natural history, diagnosis, treatment, complications, and long-term outcome. Components were scored 0 to 2 based on whether topics were addressed and in accordance with accepted practice patterns. RESULTS: Of the 124 of 150 links accessible, 84% were oriented toward laypeople. Only 43 (35%) had endorsement by a nonprofit accreditation organization, but 76 (61%) had affiliation with a medical institution/professional. Authors were mostly pediatric urologists/surgeons (36%) or unspecified (48%). Median total scores were only 16 of 44 primarily because of incomplete information rather than misinformation. High scoring components included natural history, surgical options, long-term outcomes for fertility, and malignancy. Components scoring poorly included future sexuality, imaging, hormonal use, and treatment complications. Multivariate analysis demonstrated that only accreditation status was associated with high quality (P = .05). CONCLUSIONS: Online information, when available to the public, is generally of good quality. However, many Web sites fail to adequately address all aspects of the condition. PMID- 17923208 TI - Phimotic ring topical corticoid cream (0.1% mometasone furoate) treatment in children. AB - BACKGROUND/PURPOSE: Phimosis, owing to the presence of a preputial fibrotic ring, is surgically treated in 1% of children. During the last decade, however, topical steroid treatment has been proposed for phimosis. METHODS: We present a double blind study comparing 0.1% mometasone furoate topical cream vs moisturizing cream (placebo) for the treatment of phimosis. Children aged from 2 to 13 years (n = 110) presenting with phimosis (Kikiro's classification grade 5) and scheduled for circumcision were included in this trial. The patients were evaluated after 8 weeks of topical treatment with moisturizing cream (n = 54) or steroid cream (n = 56). Nonresponders from both groups received an additional 8 weeks of steroid cream treatment. RESULTS: In the steroid group, the ring disappeared and glans exposure was obtained in 49 (88%) of 56 patients vs 28 (52%) of 54 patients in the placebo group (P < .05). After a second treatment, in the steroid group, 5 of the 7 patients were finally cured vs 22 of the 26 in the placebo group (P < .05). Two children with persisting phimosis (Kikiro's retractability grade 5 and appearance grade 3) in the steroid group (4%) vs 4 children in the placebo group (7%) ended up receiving postectomy. CONCLUSIONS: The present investigation adds up and supports the effectiveness of phimosis topical corticoid treatment. Nevertheless, hygiene and preputial traction, when appropriately performed, seem to play an important role in the disappearance of the phimotic ring as well. New studies are necessary to confirm if this is true or not. PMID- 17923209 TI - Hepaticojejunostomy and intrahepatic cystojejunostomy for type IV-A choledochal cyst. AB - BACKGROUND: The incidence of choledochal cyst with intrahepatic involvement (Todani's type IV-A cyst) is considerably high. Dilatation of the intrahepatic bile duct is frequently observed around the hepatic hilum, occasionally in the umbilical portion, and rarely in the more upstream intrahepatic bile duct, associated with or without downstream stricture. We recently encountered 2 children with type IV-A cyst associated with upstream intrahepatic ductal dilatation; one with a cystic dilatation of the medial branch arising from the left hepatic duct and another with a cyst of the medial branch arising from the anterior hepatic duct. METHODS: After excision of the extrahepatic bile duct cyst at the hilum and making a large fenestration of the intrahepatic duct cyst, hepaticojejunostomy and intrahepatic cystojejunostomy were performed using a Roux en-Y jejunal loop in both children. RESULTS: Postoperatively, both intrahepatic cysts were remarkably reduced in size, and recurrent bouts of abdominal pain did not occur for up to 4 or 5 years. CONCLUSION: Hepaticojejunostomy at the hepatic hilum, combined with intrahepatic cystojejunostomy, appears to be a recommendable procedure for an upstream intrahepatic ductal cyst of type IV-A, preventing postoperative cholangitis owing to bile stone formation. PMID- 17923210 TI - Robotic repair of a Bochdalek congenital diaphragmatic hernia in a small neonate: robotic advantages and limitations. AB - Minimally invasive repair for a Bochdalek congenital diaphragmatic hernia has been performed over the last few years with mixed results. Although the anomaly has been approached from both the abdomen and the chest, the defect can be difficult to close as the posterolateral region may be difficult to reach with precise suturing using standard rigid laparoscopic instruments. The articulating instruments of robotic surgery offer a substantial improvement in degrees of freedom and may help over come these obstacles. However, other limitations including instrument length in relation to patient size need to be accounted for when planning a robotic procedure in small children. We present a robotic repair of a foramen of Bochdalek congenital diaphragmatic in a 2.2 kg neonate using and abdominal approach with the Da Vinci Surgical Robot (Intuitive Surgical, Sunnyvale, CA). PMID- 17923211 TI - A case of congenital diaphragmatic hernia with a hernia sac attached to the liver: hints for an early embryological insult. AB - We describe here the unusual operative findings in a case of congenital diaphragmatic hernia (CDH). A neonate antenatally diagnosed as having CDH was operated via a left subcostal incision. The operative findings included a large central diaphragmatic defect of 5 x 5 cm, lined by a thick sac. The contents of the hernia included the stomach, spleen, and loops of the small and large intestine. The left side of the liver was thinned out and forming a part of the dome of the sac of the CDH. Vessels of the hepatic tissue were in continuity with the sac. There were scattered liver tissues forming cords in the sac lining. These findings were confirmed by histopathologic examinations. These findings have not been reported earlier in humans and might help to elucidate the embryology of the development of the diaphragm. Our findings suggest that this kind of defect occurs early in development, and we hypothesize that it is associated with a poorer prognosis. PMID- 17923212 TI - Parapharyngeal neuroglial heterotopia extending through the skull base in a neonate with airway obstruction. AB - We present a neonate with heterotopic nasopharyngeal brain tissue causing airway obstruction. Preoperative imaging showed extension of the mass along major neurovascular pathways into the cranial vault. Preoperative identification of intracranial extension is essential for planning surgery to prevent postoperative cerebrospinal fluid leaks or possible meningitis. PMID- 17923213 TI - Human extrahepatic portal vein obstruction correlates with decreased factor VII and protein C transcription but increased hepatocyte proliferation. AB - PURPOSE: A 3-year-old girl developed extrahepatic portal vein obstruction (EHPVO) after a liver transplant. She had sequelae of portal hypertension that required another transplantation. The circumstances allowed for comparison of liver dependent coagulation factor production between the second donor liver and the explanted liver with EHPVO. METHODS: Liver samples from the explanted first graft and the second transplant were obtained. Fresh tissue was used to perform reverse transcription-polymerase chain reaction with primers against factors V, VII, as well as VIII, protein C, and paraffin-embedded sections for hepatocyte proliferation using Ki-67 antibody as well as for apoptosis using TUNEL assay. RESULTS: The transcription of factor VII and that of protein C were decreased in the explant as compared with the newly transplanted liver (factor VII, 77% of the donor; protein C, 88% of the donor). The transcription of factor V and that of factor VIII were unchanged. The explant had a greater percentage of proliferating hepatocytes than the new organ (0.85% +/- 0.75% vs 0.11% +/- 0.21%). The percentage of apoptotic cells was similar between the 2 livers (0.09% +/- 0.13% vs 0.09% +/- 0.13%). CONCLUSIONS: Idiopathic EHPVO is associated with a reduction in liver-dependent coagulation factor transcription and an increase in hepatocyte proliferation. Portal blood flow deprivation alters hepatic homeostasis and initiates mechanisms that attempt to restore liver-dependent coagulation factors. PMID- 17923214 TI - Syndromic presentation of a pleuropulmonary blastoma associated with congenital cystic adenomatoid malformation. A case report. AB - Pleuropulmonary blastoma (PPB) is a rare malignant mesenchymal pediatric tumor with a well-recognized association with congenital cystic adenomatoid malformation (CCAM). Recently, it has been described in a patient with CCAM, multiple jejunal polyps, and cystic nephroma. We describe a similar case of a unique presentation of PPB, arising in association with CCAM and with a history of intussception caused by multiple small bowel polyps. PMID- 17923215 TI - Preoperative evaluation of the aberrant artery in intralobar pulmonary sequestration using multidetector computed tomography angiography. AB - Intralobar pulmonary sequestration is a rare congenital malformation of the lung, and identification of the aberrant artery from the systemic circulation to the sequestered lung is crucial in definitive surgery. Nowadays, various noninvasive imaging modalities such as helical computed tomography, magnetic resonance imaging, and color Doppler sonography have been developed. Among them, multidetector computed tomography angiography, which allows simultaneous imaging of the aberrant artery and venous drainage, has the potential to become the first line examination in the preoperative assessment of pulmonary sequestration. In this article, we describe 2 cases of intralobar pulmonary sequestration, in which multidetector computed tomography angiography was useful in the preoperative management and surgery was successfully performed. PMID- 17923216 TI - Laparoscopic resection of extra-adrenal pheochromocytoma--case report and review of the literature in pediatric patients. AB - Surgery for pheochromocytoma differs from that of other tumors owing to the potential release of catecholamines, which may lead to severe intraoperative hemodynamic changes. The present standard of care for resection of adrenal pheochromocytoma has become a laparoscopic approach for surgical excision. Extra adrenal pheochromocytoma is a very rare entity, especially in the pediatric age group; the utility of the laparoscopic approach is not established in this population. We present a case report of a child with hormonally active extra adrenal pheochromocytoma originating in the organ of Zuckerkandl that was resected laparoscopically. We found the laparoscopic approach gave excellent exposure, allowing for proper identification of the tumor's origin and its relation to surrounding structures; a complete resection with excellent control of the feeding blood vessels was performed. Herein we present the details of this case and a review of the relevant literature. After our initial experience we can recommend laparoscopic exploration for similar cases of suspected extra-adrenal pheochromocytoma as an appropriate tool to identify extension of the disease and estimate resectability. Proper patient preparation and monitoring are critical for success. PMID- 17923217 TI - Thoracoscopic elongation of the esophagus in long gap esophageal atresia. AB - Long gap esophageal atresia in which a primary anastomosis cannot be achieved remains a challenge. Elongation of the esophagus by traction on the 2 ends has been previously described. With the advent of thoracoscopic repair of esophageal atresia, there have thus far been no reports of thoracoscopic repair of long gap esophageal atresia. This paper describes the first successful repair of long gap esophageal atresia by thoracoscopic traction of the 2 esophageal ends and delayed thoracoscopic anastomosis. PMID- 17923218 TI - A straightforward technique for removal of the substernal bar after the Nuss operation. AB - Pectus excavatum is the most common chest wall deformity seen in the pediatric population. There have been a number of reports describing the operative correction of pectus excavatum, but scant literature is available describing removal of the substernal bar. This report describes a straightforward technique for removal of the substernal bar after the Nuss operation. This technique has been used in more than 110 patients without complications. PMID- 17923219 TI - Neonatal bronchomediastinal fistula. PMID- 17923220 TI - Congenital spigelian hernia and cryptorchidism: a distinct clinical syndrome. PMID- 17923221 TI - Re: "Is ultrasonography a good screening test for intestinal malrotation?" by Orzech et al. PMID- 17923223 TI - Mineral oil paraffins in human body fat and milk. AB - Paraffins of mineral oil origin (mineral paraffins) were analyzed in tissue fat collected from 144 volunteers with Caesarean sections as well as in milk fat from days 4 and 20 after birth of the same women living in Austria. In the tissue samples, the composition of the mineral paraffins was largely identical and consisted of an unresolved mixture of iso- and cycloalkanes, in gas chromatographic retention times ranging from n-C(17) to n-C(32) and centered at n C(23)/C(24). Since the mineral oil products we are exposed to range from much smaller to much higher molecular mass and may contain prominent n-alkanes, the contaminants in the tissue fat must be a residue from selective uptake, elimination by evaporation and metabolic degradation. Concentrations varied between 15 and 360 mg/kg fat, with an average of 60.7 mg/kg and a median of 52.5 mg/kg. Mineral paraffins might be the largest contaminant of our body, widely amounting to 1g per person and reaching 10 g in extreme cases. If food were the main source, exposure data would suggest the mineral paraffins being accumulated over many years or even lifetime. The milk samples of day 4 contained virtually the same mixture of mineral paraffins as the tissue fat at concentrations between 10 and 355 mg/kg (average, 44.6 mg/kg; median, 30 mg/kg). The fats from the day 20 milks contained <5-285 mg/kg mineral paraffins (average, 21.7; median, 10mg/kg), whereby almost all elevated concentrations were linked with a modified composition, suggesting a new source, such as the use of breast salves. The contamination of the milk fat with mineral paraffins seems to decrease more rapidly than for other organic contaminants, and the transfer of mineral paraffins to the baby amounts to only around 1% of that in the body of the mother. PMID- 17923224 TI - Alpha-Tocopheryl succinate: toxicity and lack of anti-tumour activity in immuno competent mice. AB - Alpha-tocopheryl succinate (alpha-TOS), an analogue of vitamin E (VitE), inhibits peritoneal human malignant mesoethelioma xenograft development in immuno compromised mice via the induction of apoptosis of tumour cells [Tomasetti, M., Gellert, N., Procopio, A., Neuzil, J., 2004. A vitamin E analogue suppresses malignant mesothelioma in a preclinical model: a future drug against a fatal neoplastic disease? Int. J. Cancer 109, 641-642]. We tested the effect of systemic alpha-TOS treatment in our immuno-competent and syngeneic murine mesothelioma model. VitE analogues such as alpha-TOS have been developed for clinical use as supplements mainly for the treatment of VitE deficiency and are considered safe and non-toxic when taken orally. In our murine model of mesothelioma alpha-TOS was not only ineffective at inhibiting established tumour development at the published doses, but resulted in severe side effects characterized by both behavioural changes, intra-peritoneal abnormalities and the destruction of T cells. Toxicity of alpha-TOS has not been reported to date perhaps due to a lack of studies conducted in fully immuno-competent hosts. Our results suggest that the translation of animal studies to clinical treatment with alpha-TOS requires careful consideration. PMID- 17923225 TI - Effects of conditioners on microshear bond strength to enamel after carbamide peroxide bleaching and/or casein phosphopeptide-amorphous calcium phosphate (CPP ACP) treatment. AB - OBJECTIVES: To evaluate (a) the enamel microshear bond strength (MSBS) of a universal adhesive and (b) the effects of conditioning with a self-etching primer adhesive with/without prior bleaching and/or casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) application. METHODS: Thirty-five molars were cut into four sections, assigned randomly to four groups (no treatment; 16% carbamide peroxide bleaching; CPP-ACP-containing paste (Tooth Mousse, TM); bleaching and TM) and treated accordingly. Specimens were divided into two for bonding with either a self-etching primer (Clearfil SE Bond, CSE) or a total-etch adhesive (Single Bond, SB). Specimens for CSE bonding were subdivided for one of four preconditioning treatments (no conditioning; 30-40% phosphoric acid (PA); 15% EDTA; 20% polyacrylic acid conditioner (Cavity conditioner, CC) and treated. The adhesives were applied and resin composite bonded to the enamel using microtubes (internal diameter 0.75mm). Bonds were stressed in shear until failure, mean MSBS calculated and data analysed using ANOVA with Tukey's HSD test (alpha=0.05). The modes of bond failure were assessed and classified. RESULTS: Two-way ANOVA revealed significant differences between treatments (P<0.0001), conditioners (P<0.0001) and a significant interaction between treatments and conditioners (P=0.001). One-way ANOVA revealed no significant differences in MSBS following any of the treatments for SB; following TM application for CSE without preconditioning; and significant differences in MSBS following bleaching with and without TM application for CSE. With preconditioning, applying PA before CSE post bleaching and either PA or CC before CSE post-TM application, resulted in significant differences in MSBS (P<0.05). CONCLUSIONS: The use of conditioners prior to bonding with the self-etching primer adhesive system on treated enamel may significantly improve bond strengths. PMID- 17923228 TI - In vivo deletion analysis of the architecture of a multiprotein complex of translation initiation factors. AB - Protein complexes play a critical role in virtually all cellular processes that have been studied to date. Comprehensive knowledge of the architecture of a protein complex of interest is, therefore, an important prerequisite for understanding its role in the context of a particular pathway in which it participates. One of the possible approaches that has proven very useful in characterizing a protein complex is outlined in this chapter using the example of the eukaryotic initiation factor 3 (eIF3) and some of its binding partners. eIF3 is one of the major players in the translation initiation pathway because it orchestrates several crucial steps that ultimately conclude with formation of the 80S ribosome where the anticodon of methionyl-tRNA(i)(Met) base-pairs with the AUG start codon of the mRNA in the ribosomal P-site. We previously demonstrated that, in the budding yeast Saccharomyces cerevisiae, eIF3 closely cooperates with several other eIFs to stimulate recruitment of methionyl-tRNA(i)(Met) and mRNA to the 40S ribosome and that it forms, together with eIFs 1, 2, and 5, an important intermediate in translation initiation called the multifactor complex (MFC). Here, we summarize the fundamental procedure that allowed in-depth characterization of the MFC composition and identification of protein-protein interactions among its constituents. Primarily, we describe in detail in vivo purification techniques that, in combination with systematic deletion analysis, produced a 3D subunit interaction model for the MFC. Site-directed clustered-10 alanine-mutagenesis (CAM) employed to investigate the physiological significance of individual interactions is also presented. The general character of the entire procedure makes it usable for first-order structural characterization of virtually any soluble protein complex in yeast. PMID- 17923227 TI - Purification of FLAG-tagged eukaryotic initiation factor 2B complexes, subcomplexes, and fragments from Saccharomyces cerevisiae. AB - The eukaryotic initiation factor 2B (eIF2B) is a five-subunit guanine nucleotide exchange factor, that functions during translation initiation to catalyze the otherwise slow exchange of GDP for GTP on its substrate eIF2. Assays to measure substrate interaction and guanine nucleotide release ability of eIF2B require the complex to be purified free of interacting proteins. We have also found that a subcomplex of two subunits, gamma and epsilon or the largest one, epsilon alone, promotes this activity. Within eIF2Bepsilon, the catalytic center requires the C terminal 200 residues only. Here, we describe our protocols for purifying the Saccharomyces cerevisiae eIF2B complexes and the catalytic subunit using FLAG tagged proteins overexpressed in yeast cells. Using commercially available FLAG affinity resin and high salt buffer, we are able to purify active eIF2B virtually free of contaminants. PMID- 17923229 TI - An approach to studying the localization and dynamics of eukaryotic translation factors in live yeast cells. AB - The discovery of Green Fluorescent Protein (GFP) and the development of technology that allows specific proteins to be tagged with GFP has fundamentally altered the types of question that can be asked using cell biological methods. It is now possible not only to study where a protein is within a cell, but also feasible to study the precise dynamics of protein movement within living cells. We have exploited these technical developments and applied them to the study of translation initiation factors in yeast, focusing particularly on the key regulated guanine nucleotide exchange step involving eIF2B and eIF2. This chapter summarizes current methodologies for the tagging and visualization of GFP-tagged proteins involved in translation initiation in live yeast cells. PMID- 17923222 TI - Top-down predictions in the cognitive brain. AB - The human brain is not a passive organ simply waiting to be activated by external stimuli. Instead, we propose that the brain continuously employs memory of past experiences to interpret sensory information and predict the immediately relevant future. The basic elements of this proposal include analogical mapping, associative representations and the generation of predictions. This review concentrates on visual recognition as the model system for developing and testing ideas about the role and mechanisms of top-down predictions in the brain. We cover relevant behavioral, computational and neural aspects, explore links to emotion and action preparation, and consider clinical implications for schizophrenia and dyslexia. We then discuss the extension of the general principles of this proposal to other cognitive domains. PMID- 17923230 TI - In vitro and tissue culture methods for analysis of translation initiation on the endoplasmic reticulum. AB - For mRNAs encoding secretory and integral membrane proteins, translation initiation is thought to begin a process of mRNA localization where mRNA/ribosome/nascent chain complexes (RNCs) are trafficked from the cytosol compartment to the endoplasmic reticulum (ER). At the ER membrane, RNCs bind to a protein-conducting channel via the large ribosomal subunit and protein translocation ensues through coupling of the ribosomal nascent protein exit site with the protein-conducting channel. At the termination of translation, ribosomal subunits are thought to dissociate from the ER to return to a common cytoplasmic pool and participate in additional cycles of initiation, translation, targeting, termination, and ER membrane release. Experimental evidence has demonstrated that ER-membrane ribosomes are capable of de novo initiation, that mRNA partitioning to the ER membrane does not, per se, require translation of an encoded signal sequence, and that ribosomal subunit dissociation from the ER membrane is not obligatorily coupled to protein synthesis termination. These findings suggest that the cycle of protein synthesis-initiation, elongation, and termination-can occur on the two-dimensional plane of the ER membrane and challenge current views on the subcellular restriction of translation initiation to the cytosol, the role of the ribosome cycle in partitioning mRNA between the cytosol and ER, and the in vivo basis for termination-induced ribosomal subunit dissociation. In the following chapter, we provide detailed experimental methods to study protein synthesis initiation on the ER membrane. PMID- 17923231 TI - Mammalian stress granules and processing bodies. AB - The packaging of cytoplasmic mRNA into discrete RNA granules regulates gene expression by delaying the translation of specific transcripts. Specialized RNA granules found in germ cells direct the timing of maternal mRNA translation to promote germ cell development in the early embryo and establish the germ line for the next generation. Similarly, select neuronal mRNA transcripts are packaged into translationally inert RNA granules, transported to sites where their protein products are required, and only then activated and translated. Following translation, however, newly inactivated mRNAs released from polysomes can also be packaged into dynamic, transient structures known as stress granules (SGs) and processing bodies (PBs). Stress granules are composed largely of stalled preinitiation complexes, and contain mRNA, small ribosomal subunits, eIF3, eIF4E, eIF4G, and PABP, as their core components. PBs are associated with mRNA decay and contain the decapping enzymes DCP1/2, the 5' to 3' exonuclease Xrn1, the Lsm proteins (1-7), and the scaffolding proteins hedls/GE-1 and GW182. Both SGs and PBs contain mRNA, eIF4E, microRNAs and argonaute proteins, and various regulators of mRNA stability and translation (TTP, RCK/p54, and CPEB). Thus, SGs and PBs share some protein and mRNA components, but also contain a number of unique markers specific to each structure. We describe markers and staining procedures used to identify these distinct types of RNA granules, describe conditions that promote their assembly and disassembly, and establish YB-1 as a useful marker of SGs and PBs. PMID- 17923232 TI - Methods to analyze microRNA-mediated control of mRNA translation. AB - MicroRNAs (miRs) are an important class of gene regulators that affect a wide range of biological processes. Despite the early recognition of miRs as translational regulators and intense interest in studying this phenomenon, it has so far not been possible to derive a consensus model for the underlying molecular mechanism(s). The potential of miRs to act in a combinatorial manner and to also promote mRNA decay creates conceptual and technical challenges in their study. Here, we discuss critical parameters in design and analysis of experiments used to study miR function including creation of synthetic miR and mRNA partners for assay of translational inhibition using luciferase reporters; measurement of mRNA stability after miR action; defining poly(A) tail length in miR target mRNA; determining the distribution of miRs and their target mRNAs in polysome profiles; and visualization of P-body components. We describe protocols for each of these procedures. PMID- 17923233 TI - Methods for studying signal-dependent regulation of translation factor activity. AB - The translational machinery of mammalian cells is regulated through the phosphorylation of a number of its components, especially translation factor proteins. These include factors involved in the initiation and elongation stages of translation, and proteins that modify their activity. Examples include eukaryotic initiation factor (eIF) 4E, eukaryotic elongation factor (eEF) 2, and eIF4E-binding protein 1 (4E-BP1). Their phosphorylation is mediated by protein kinases that, in turn, are regulated by specific intracellular signaling pathways. These pathways include those mediated via the mammalian target of rapamycin (mTOR), the ERK and p38 MAP kinase pathways, and protein kinase B (Akt). These pathways are activated by hormones (e.g., insulin), growth factors, mitogens, and other extracellular stimuli. In some cases, amino acids also modulate the pathway (e.g., mTOR). Procedures are described for determining the states of phosphorylation and/or activity of several translation factors, and of kinases that phosphorylate them. We also outline procedures for assessing the states of activation of relevant signaling pathways. In addition, we provide guidelines on using small molecule inhibitors to assess the involvement of specific signaling pathways in controlling translation factors and protein synthesis. PMID- 17923234 TI - Analysis of mRNA translation in cultured hippocampal neurons. AB - Synaptic plasticity, the ability of neuronal synapses to undergo morphological and biochemical changes in response to various stimuli, forms the underlying basis of long-term memory storage. Regulated mRNA translation at synapses is required for this plasticity. However, the mechanism by which translation at synapses is controlled and how the encoded proteins modulate persistent changes in synaptic morphology and functional integration in response to different input stimulations remain mostly unclear (Schuman et al., 2006; Sutton and Schuman, 2006). One approach to investigating the relationship between protein synthesis and plasticity is to identify factors, such as RNA binding proteins that control translation in the neurons and then determine the identities of the mRNAs to which they are bound. Molecular and cellular techniques have been employed in cultured neurons to study sequence-specific RNA-binding proteins, for example, the Cytoplasmic Polyadenylation Element Binding protein (CPEB) (Huang et al., 2002, 2003) and the Fragile-X Mental Retardation Protein (FMRP) (Vanderklish and Edelman, 2005; Zalfa et al., 2006) for their functions in localizing and regulating translation of mRNAs. Although several CPE-containing neuronal RNAs that undergo activity-dependent polyadenylation (Du and Richter, 2005; Wu et al., 1998) and FMRP-interacting mRNAs have been identified (Brown et al., 2001; Miyashiro et al., 2003), the validation of these targets whose translation is important for plasticity in vivo remains to be demonstrated (Darnell et al., 2005). In general, primary neurons in culture are difficult to manipulate. For example, they do not proliferate and their transfection efficiency is low ( approximately 1 to 10% of cells); this low efficiency is reduced even further as the cells age in culture, which hampers their practical use for biochemical analysis. When biochemical approaches are applied, they are often carried out in other more facile model systems, such as oocytes, in the case of CPEB, or in brains derived from knockout mice, for both CPEB and FMRP. However, the development of various viral delivery systems, shRNA knockdown techniques, reporter assays with high sensitivity, and neuron culture protocols have allowed investigators to analyze translational control in these cells, which may ultimately be used to investigate key mechanisms of synaptic plasticity. We have employed these procedures to investigate the function of CPEB3, a novel RNA binding protein, in primary rat hippocampal neurons (Huang et al., 2006); here, we describe the experimental details of our methods, which could be used for any RNA binding protein. PMID- 17923235 TI - Detecting ribosomal association with the 5' leader of mRNAs by Ribosome Density Mapping (RDM). AB - In eukaryotes, scanning of the 5' leader by the small ribosomal subunit precedes recognition of the start codon. Thus, various sequence elements that are located within this region may affect ribosomes' progression and lead to significant effects on translation. Most notable are short ORFs located upstream of the start codon, which are known to regulate the translation of the main ORF in the transcript. The function of these elements is likely to correlate with altered ribosomal association with the 5' leader of the mRNA. Currently, the only method to determine the ribosomal association of different regions of the mRNA in vivo is the Ribosome Density Mapping (RDM) procedure. This method entails cleavage of the target mRNA by specific oligodeoxynucleotides and RNase H and separation of the cleavage products by velocity sedimentation in a sucrose gradient. In this chapter, we provide a detailed protocol for this procedure and discuss its feasibility. PMID- 17923236 TI - Genome-wide analysis of mRNA polysomal profiles with spotted DNA microarrays. AB - The sedimentation of an mRNA in sucrose gradients is highly affected by its ribosomal association. Sedimentation analysis has therefore become routine for studying changes in ribosomal association of mRNAs of interest. DNA microarray technology has been combined with sedimentation analysis to characterize changes in ribosomal association for thousands of mRNAs in parallel. Such analyses revealed mRNAs that are translationally regulated and have provided new insights into the translation process. In this chapter, we describe possible experimental designs for analyzing genome-wide changes in ribosomal association, and discuss some of their advantages and disadvantages. We then provide a detailed protocol for analysis of polysomal fractions using spotted DNA microarrays. PMID- 17923238 TI - Methods for identifying compounds that specifically target translation. AB - This chapter presents methods and protocols suitable for the identification and characterization of inhibitors of the prokaryotic and/or eukaryotic translational apparatus as a whole or targeting specific, underexploited targets of the bacterial protein synthetic machinery such as translation initiation and aminoacylation. Some of the methods described have been used successfully for the high-throughput screening of libraries of natural or synthetic compounds and make use of model "universal" mRNAs that can be translated with similar efficiency by cellfree extracts of bacterial, yeast, and HeLa cells. Other methods presented here are suitable for secondary screening tests aimed at identifying a specific target of an antibiotic within the translational pathway of prokaryotic cells. PMID- 17923237 TI - Synthesis of anti-reverse cap analogs (ARCAs) and their applications in mRNA translation and stability. AB - Synthetic capped RNA transcripts produced by in vitro transcription in the presence of m(7)Gp(3)G have found a wide application in studying such processes as mRNA translation, pre-mRNA splicing, mRNA turnover, and intracellular transport of mRNA and snRNA. However, because of the presence of a 3'-OH on both m(7)Guo and Guo moieties of the cap structure, one-third to one-half of the mRNAs contain a cap incorporated in the reverse orientation. The reverse cap structures bind poorly to eIF4E, the cap binding protein, and reduce overall translational efficiency. We therefore replaced the conventional m(7)Gp(3)G cap by "anti reverse" cap analogs (ARCAs) in which the 3'-OH of m(7)Guo moiety was substituted by 3'-deoxy or 3'-O-methyl groups, leading to m(7)3'dGp(3)G or m(2)(7,3'-O) Gp(3)G, respectively. The class of ARCAs was extended to analogs possessing an O methyl group or deoxy group at C2' of m(7)Guo. We have also developed a series of ARCAs containing tetra- and pentaphosphates. mRNAs capped with various ARCAs were translated 1.1- to 2.6-fold more efficiently than their counterparts capped with m(7)Gp(3)G in both in vitro and in vivo systems. In a separate series, a methylene group was introduced between the alpha- and beta-, or beta- and gamma phosphate moieties, leading to m(2)(7,3'-O)Gpp(CH2)pG and m(2)(7,3'-O)Gp(CH2)ppG. These analogs are resistant to cleavage by the decapping enzymes Dcp1/Dcp2 and DcpS, respectively. mRNA transcripts capped with m(2)(7,3'-O)Gpp(CH2)pG were more stable when introduced into cultured mammalian cells. In this chapter, we describe the synthesis of representative ARCAs and their biophysical and biochemical characterization, with emphasis on practical applications in mRNA translation. PMID- 17923239 TI - Identifying small molecule inhibitors of eukaryotic translation initiation. AB - In eukaryotes, translation initiation is rate-limiting with much regulation exerted at the ribosome recruitment and ternary complex (eIF2.GTP.Met tRNA(i)(Met)) formation steps. Although small molecule inhibitors have been extremely useful for chemically dissecting translation, there is a dearth of compounds available to study the initiation phase in vitro and in vivo. In this chapter, we describe reverse and forward chemical genetic screens developed to identify new inhibitors of translation. The ability to manipulate cell extracts biochemically, and to compare the activity of small molecules on translation of mRNA templates that differ in their factor requirements for ribosome recruitment, facilitates identification of the relevant target. PMID- 17923240 TI - Isolation and identification of eukaryotic initiation factor 4A as a molecular target for the marine natural product Pateamine A. AB - Natural products continue to demonstrate their utility both as therapeutics and as molecular probes for the discovery and mechanistic deconvolution of various cellular processes. However, this utility is dampened by the inherent difficulties involved in isolating and characterizing new bioactive natural products, in obtaining sufficient quantities of purified compound for further biological studies, and in developing bioactive probes. Key to characterizing the biological activity of natural products is the identification of the molecular target(s) within the cell. The marine sponge-derived natural product Pateamine A (PatA) has been found to be an inhibitor of eukaryotic translation initiation. Herein, we describe the methods utilized for identification of the eukaryotic translation initiation factor 4A (eIF4A) as one of the primary protein targets of PatA. We begin by describing the synthesis of an active biotin conjugate of PatA (B-PatA), made possible by total synthesis, followed by its use for affinity purification of PatA binding proteins from cellular lysates. We have attempted to present the methodology as a general technique for the identification of protein targets for small molecules including natural products. PMID- 17923242 TI - Bone marrow-derived CD8alpha+TCR- cells that facilitate allogeneic bone marrow transplantation are a mixed population of lymphoid and myeloid progenitors. AB - OBJECTIVE: We have characterized a hematopoietic cell population isolated from murine bone marrow that can facilitate purified hematopoietic stem cell engraftment across fully allogeneic major histocompatibility complex barriers. These facilitating cells (FCs) are classically identified as CD8alpha(+)TCR(-) by flow cytometry. Prior work has demonstrated that FCs are comprised of a heterogeneous cell population with both lymphoid and myeloid phenotypes. The present investigation was designed to more precisely characterize these subsets in terms of both phenotype and developmental potential. METHODS: Using fluorescence-activated cell sorting analysis, freshly isolated FCs were characterized for phenotypic expression of various lymphocyte progenitor markers. The lymphopoietic potential of FCs was evaluated by culturing freshly isolated FCs on bone marrow stroma cells overexpressing notch ligand 1 (OP9-DL1). Transcripts specific to pTalpha and TCRalpha were quantitated by employing real time reverse transcription polymerase chain reaction. Maturation of the T-cell receptor (TCR) on FCs was biochemically analyzed by immunoprecipitation. RESULT: Freshly isolated FCs had significant expression of CD44(+)CD25(-) and CD44(+)CD25(+) phenotypes. A discrete subset of CD8(+)CD4(+) cells are also identified in the FC population, similar to the double-positive phase of thymocyte development. Of particular interest, FCs express pre-TCRalpha (pTalpha) mRNA and protein as demonstrated by reverse transcription polymerase chain reaction, intracellular staining and immunoprecipitation. FCs grown on OP9-DL1 with interleukin-7 and FMS-like tyrosine kinase 3 ligand can mature into CD44( )CD25(+), CD8(+)CD4(+) and CD8(+) T cells. During this developmental process, expression of the 33-kDa pTalpha chain was replaced by a mature 40-kDa TCRalpha chain. CONCLUSION: Taken together, these data demonstrate for the first time that the marrow-derived FC contains a T-cell progenitor population that closely resembles developing thymocytes. PMID- 17923241 TI - Marital status and variability in cortisol excretion in postmenopausal women. AB - Based on the premise that acute and chronic stresses stimulate and suppress cortisol secretion, respectively, and the hypothesis that marriage provides a buffer to stress, we tested whether extreme values of serum cortisol concentrations would be less likely in married women than in unmarried women. Three hundred women were recruited from two central Connecticut communities. Cortisol was measured in overnight urine samples using liquid chromatography tandem mass spectrometry. Information on each subject's demographic characteristics, such as income and education level was collected. Mean log urinary cortisol was virtually identical in married and unmarried women, however, as predicted, the variance was significantly larger in the unmarried group (p=0.01). After adjustment for potential confounders, multivariate logistic regression still revealed that absolute deviation of log(10) cortisol from the mean was smaller for married versus unmarried women (p<0.01); deviation from the mean cortisol was also higher for non-working than working women. These results support the idea that marriage and employment reduce the extreme levels of cortisol secretion, and by extension, this may reflect differences in levels of stress in married and in working women compared to unmarried and non-working women. PMID- 17923243 TI - Angiopoietin-1 supports induction of hematopoietic activity in human CD34- bone marrow cells. AB - OBJECTIVE: Hematopoietic stem cells (HSCs) consist of heterogenous subpopulations, one of which is CD34(-) HSCs. Recent development of successful engraftment by intra-bone marrow transplantation revealed severe combined immunodeficiency (scid) mouse-repopulating cell (SRC) activity in human CD34(-) cord blood (CB) cells. On the other hand, CD34(-) cells from bone marrow (BM) cells remain relatively undefined. Here, we investigated pre-SRC populations in human BM CD34(-) cells and the effect of the niche-related factor, angiopoietin 1, on them. METHODS: Two populations in BM CD34(-) cells (namely M cells and S cells) were purified by flow cytometry. Then, they were cocultured with six growth factors on the hematopoietic-supportive mouse BM stromal cell line, HESS-5 or AHESS-5 that were engineered to produce human angiopoietin-1, because we detected Tie2 expression on M cells and S cells. Cultured cells were assessed for their in vitro and in vivo hematopietic activities. RESULTS: After 7 days in coculture, AHESS-5 was stronger more effective than HESS-5 in converting M and S cells to CD34(+) cells (M cells: 67.4% vs 17.5%, n =6, p < 0.001) (S cells: 42.3% vs 2.3%, n = 6, p < 0.001). Furthermore, both M and S cells were able to engraft in immunodeficient mice after they were cocultured on AHESS-5. CONCLUSIONS: Results suggest that angiopoietin-1 supports SRC activities in human CD34(-) BM cells, as murine studies demonstrated. Furthermore, identification of previously undetected subpopulations of BM CD34(-) HSCs unveils heterogenous components in the stem cell pool. PMID- 17923244 TI - Evidence that intracoronary-injected CD133+ peripheral blood progenitor cells home to the myocardium in chronic postinfarction heart failure. AB - OBJECTIVE: To study the biodistribution of purified CD133(+) cells after intracoronary injection in patients with stable chronic postinfarction heart failure. PATIENTS AND METHODS: Patients with longstanding myocardial infarction (>12 months prior to inclusion) and with an accessible left coronary artery were eligible. CD133(+) cells were mobilized with granulocyte colony-stimulating factor and purified with a CliniMACS device. Cells were labeled with (111)Indium and injected through a balloon catheter in a coronary artery feeding the necrotic or viable infarct-related region of the left ventricle during a standard coronary catheterization procedure. The total body biodistribution of (111)Indium was studied with a dual-head gamma camera in combination with (99m)Technetium sestaMIBI cardiac distribution analysis. RESULTS: The number of CD133(+) cells injected ranged between 5 and 10 x 10(6) cells (low dose, three patients) or between 18.5 and 50 x 10(6) cells (high dose, five patients). In the five patients receiving the higher cell doses, a clear residual radioactivity was observed at the level of the chronic injury at 2, 12, and up to 36 hours after injection. A detailed analysis in two patients showed 6.9% to 8.0% (after 2 hours) and 2.3% to 3.2% (after 12 hours) residual radioactivity at the heart. No adverse events were observed during the procedure and up to 3 months follow-up. CONCLUSIONS: We demonstrate that CD133(+) progenitor cells are capable of homing to the postinfarction remodeling myocardium after intracoronary injections in patients with chronic postinfarction heart failure. PMID- 17923245 TI - NF-E2-mediated enhancement of megakaryocytic differentiation and platelet production in vitro and in vivo. AB - OBJECTIVE: NF-E2 is a prime regulator of megakaryocyte (MK) terminal differentiation and platelet release. By overexpressing the p45 subunit of NF-E2, we aim to increase the proportion of mature MKs and the potential for platelet production in vitro and in vivo. METHODS: Retroviral vectors expressing p45-NF-E2 together with the enhanced green fluorescent protein (eGFP) were used to transduce murine bone marrow cells (BMCs). Aspects of MK differentiation, proliferation, proplatelet, and platelet production were evaluated. RESULTS: Compared to controls, a higher proportion of BMCs overexpressing p45-NF-E2 were found to express the MK markers CD41, CD42a, and CD42b, with some effect on cell proliferation. Early MK differentiation, characterized by colony-forming unit (CFU)-MK formation, was enhanced by p45-NF-E2 overexpression at the expense of CFU-granulocyte macrophage development. An increased number of acetylcholinesterase(+) MKs was also observed in NF-E2(++) cultures. Although endomitosis was found not to be affected, the resultant upregulation of NF-E2 target genes was also followed by significant increases in proplatelet and functional platelet production. Transplantation of enriched MK progenitor cells overexpressing p45-NF-E2 into lethally irradiated mice resulted in a threefold increase in eGFP(+)/NF-E2(++) platelet production in vivo over 10 days, although no appreciable expansion in their number was observed over 32 days. CONCLUSION: These results suggest that enforced expression of p45-NF-E2 selectively enhances many aspects of MK differentiation, including MK maturation, proplatelet formation, and platelet release. In addition, p45 overexpression increases MK commitment during early megakaryopoiesis, while inhibiting white blood cell differentiation. PMID- 17923246 TI - P-glycoprotein is downregulated in KG1a-primitive leukemia cells by LDL cholesterol deprivation and by HMG-CoA reductase inhibitors. AB - OBJECTIVE: P-glycoprotein (pgp) is a membrane transporter encoded by the multidrug resistance (MDR1, ABCB1) gene. Pgp is a poor prognostic factor in elderly patients with acute myeloid leukemia (AML). In addition to its role in drug efflux, pgp has been implicated in cellular cholesterol homeostasis. We investigated the effects of exogenous cholesterol removal on pgp expression and function. METHODS: KG1a drug-naive, primitive leukemia cells were cultured in serum-free medium with or without the addition of low-density lipoprotein (LDL) cholesterol. After 72 hours, pgp expression and function was assessed by flow cytometry and total cholesterol content of the KG1a cells was determined by the Amplex Red cholesterol assay. The addition of clinically available cholesterol lowering agents, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors to KG1a cells was also assessed. RESULTS: There was a 39% (SEM = 8.3%; p = 0.03) decrease in pgp protein expression after 3 days of serum-free culture. The decrease was also observed at the message and functional levels. In the presence of low-density lipoprotein cholesterol, pgp expression was restored to 86% of the basal value. Addition of a HMG-CoA reductase inhibitor to KG1a cells resulted in an additional 26% (lovastatin, p = 0.03) and 16% (pravastatin, p = 0.05) reduction in pgp, respectively. Furthermore, toxicity of the pgp substrate drug daunorubicin was enhanced following lovastatin preculture (p = 0.04). CONCLUSION: LDL cholesterol contributes to pgp expression and chemoresistance in primitive leukemia cells. Use of HMG-CoA reductase inhibitors may be of clinical value in lowering pgp expression in AML. PMID- 17923247 TI - Scenario-based stakeholder engagement: incorporating stakeholders preferences into coastal planning for climate change. AB - Climate change poses many challenges for ecosystem and resource management. In particular, coastal planners are struggling to find ways to prepare for the potential impacts of future climate change while dealing with immediate pressures. Decisions on how to respond to future risks are complicated by the long time horizons and the uncertainty associated with the distribution of impacts. Existing coastal zone management approaches in the UK either do not adequately incorporate changing stakeholder preferences, or effectively ensure that stakeholders are aware of the trade-offs inherent in any coastal management decision. Using a novel method, scenario-based stakeholder engagement, which brings together stakeholder analysis, climate change management scenarios and deliberative techniques, the necessary trade-offs associated with long term coastal planning are explored. The method is applied to two case studies of coastal planning in Christchurch Bay on the south coast of England and the Orkney Islands off the north coast of Scotland. A range of conflicting preferences exist on the ideal governance structure to manage the coast under different climate change scenarios. In addition, the results show that public understanding of the trade-offs that have to be made is critical in gaining some degree of public support for long term coastal decision-making. We conclude that scenario-based stakeholder engagement is a useful tool to facilitate coastal management planning that takes into account the complexities and challenges of climate change, and could be used in conjunction with existing approaches such as the Shoreline Management Planning process. PMID- 17923248 TI - Intensive olive orchards on sloping land: good water and pest management are essential. AB - There is intensive cultivation of olives on sloping land in Jaen-Granada (Spain), Basilicata (Italy) and Western Crete (Greece). The intensive olive groves here are characterised by a tree density of about 250treesha(-1), yearly fertilisation and pruning, several chemical sprays for pest control, soil tillage once to thrice per year and irrigation up to 2700m3ha(-1)yr(-1). Intensive management results in high yields of 3600-6500kgha(-1) but also higher labour costs of 1154 1590euroha(-1)yr(-1), varying per area. The major environmental concerns in this system are related to chemical residues in the fruit, the extinction of useful insects, the depletion of groundwater resources, the pollution of soil and water and the erosion of soil. This paper describes the impact of intensive orchard management on natural resources and gives recommendations for soil and water conservation, reduction of chemicals use and biodiversity enhancement. The specific recommendations for the relevant stakeholders--farmers, technicians, agricultural services and policy makers--are based on the experimental evaluation of different agricultural practices and a socio-economic analysis of local and global production and markets. PMID- 17923249 TI - Multireservoir real-time operations for flood control using balanced water level index method. AB - This paper presents a real-time simulation-optimization operation procedure for determining the reservoir releases at each time step during a flood. The proposed procedure involves two models, i.e., a hydrological forecasting model and a reservoir operation model. In the reservoir operation model, this paper compares two flood-control operation strategies for a multipurpose multireservoir system. While Strategy 1 is the real-time joint reservoir operations without using the balanced water level index (BWLI) method, Strategy 2 involves real-time joint reservoir operations using the BWLI method. The two strategies presented are formulated as mixed-integer linear programming (MILP) problems. The idea of using the BWLI method is derived from the HEC-5 program developed by the US Army Corps of Engineers. The proposed procedure has been applied to the Tanshui River Basin system in Taiwan using the 6h ahead forecast data of six typhoons. A comparison of the results obtained from the two strategies reveals that Strategy 2 performs much better than Strategy 1 in determining the reservoir real-time releases throughout the system during flood emergencies in order to minimize flooding, while maintaining all reservoirs in the system in balance if possible. Consequently, the proposed model using the BWLI method demonstrates its effectiveness in estimating real-time releases. PMID- 17923255 TI - Tissue engineering and reconstructive surgery in urology. PMID- 17923250 TI - Traditional olive orchards on sloping land: sustainability or abandonment? AB - Traditional olive orchards account for a large share of the area under olives in the EU, particularly in marginal areas, like those analysed in the OLIVERO project. In general, traditional olive growing can be described as a low intensity production system, associated with old (sometimes very old) trees, grown at a low density, giving small yields and receiving low inputs of labour and materials. Though such systems are environmentally sustainable, their economic viability has become an issue, since EU policies favour more intensive and competitive systems. Orchards that have not been intensified seem to be threatened by the recent reform of the EU olive and olive oil policy, as income support has been decoupled from production. The main purpose of this paper is to identify the present constraints to traditional olive growing, and to recommend some private and public interventions to prevent its abandonment. During the OLIVERO project, traditional olive production systems were identified and described in five target areas (Tras-os-Montes--Portugal, Cordoba and Granada/Jaen--Spain, Basilicata/Salerno--Italy, and West Crete--Greece). The causes and consequences of abandonment are discussed, based on the analysis of the costs and returns, which revealed that these systems are barely economically sustainable. Their viability is only assured if reduced opportunity costs for family labour are accepted, and the olive growing is part-time. Based on these results, recommendations are made to prevent the abandonment of traditional olive growing and to preserve its environmental benefits. PMID- 17923256 TI - Investigations into the design principles in the chemotactic behavior of Escherichia coli. AB - Inspired by the recent studies on the analysis of biased random walk behavior of Escherichia coli[Passino, K.M., 2002. Biomimicry of bacterial foraging for distributed optimization and control. IEEE Control Syst. Mag. 22 (3), 52-67; Passino, K.M., 2005. Biomimicry for Optimization, Control and Automation. Springer-Verlag, pp. 768-798; Liu, Y., Passino, K.M., 2002. Biomimicry of social foraging bacteria for distributed optimization: models, principles, and emergent behaviors. J. Optim. Theory Appl. 115 (3), 603-628], we have developed a model describing the motile behavior of E. coli by specifying some simple rules on the chemotaxis. Based on this model, we have analyzed the role of some key parameters involved in the chemotactic behavior to unravel the underlying design principles. By investigating the target tracking capability of E. coli in a maze through computer simulations, we found that E. coli clusters can be controlled as target trackers in a complex micro-scale-environment. In addition, we have explored the dynamical characteristics of this target tracking mechanism through perturbation of parameters under noisy environments. It turns out that the E. coli chemotaxis mechanism might be designed such that it is sensitive enough to efficiently track the target and also robust enough to overcome environmental noises. PMID- 17923258 TI - Diffuse cortical lesions with hemorrhage in cerebral Whipple's disease. AB - A 30-year-old Chinese male with a history of diarrhea and arthralgia presented for evaluation of progressive dementia, epilepsy, and increased intracranial pressure. Imaging of the brain showed progressive cortical and subcortical lesions with hemorrhage involving the bilateral temporal and occipital lobes, the posterior parietal lobes, and the left frontal lobe. "Foamy" periodic acid-Schiff (PAS)-positive macrophages were demonstrated on brain biopsy. The patient showed clinical improvement following treatment with chloromycetin and sulfadiazine for 2 months. This constitutes the first reported case of cerebral Whipple's disease with diffuse cortical lesions with hemorrhage reported in a Chinese individual. Further, this case points out the significance of early recognition and treatment of cerebral Whipple's disease, especially in those cases with unusual manifestations. PMID- 17923257 TI - 17beta-Estradiol utilizes the estrogen receptor to regulate CD16 expression in monocytes. AB - Estrogen can significantly influence CD16 expression and alter monocytic cytokine release upon CD16 receptor activation. However, the function of the estrogen receptor (ER) alpha and beta in this response is unclear. To test whether estrogen binds ERalpha and/or ERbeta to affect CD16 expression, monocytic cells were treated with and without physiological levels of 17beta-estradiol and various doses of the ERalpha and ERbeta antagonist fulvestrant followed by measurement of CD16 transcript levels. To determine how estrogen induced changes in TNF-alpha and IL-1beta release due to CD16 activation we quantitated the amount of cytokines after treatment with estrogen, fulvestrant and antibodies that specifically bind and activate the CD16 receptor. Interaction of ERalpha and the CD16 promoter was then determined by chromatin immunoprecipitation. Furthermore, specific promoter elements utilized by estrogen to control CD16 expression were mutated and expression from a luciferase reporter quantitated after transfection. Using the luciferase reporter construct containing a wild type CD16 promoter, the role of ERalpha and ERbeta in the estrogen response was tested by treating transfected monocytes with an ERalpha specific agonist or an ERbeta specific agonist and measuring expression. Our results show that CD16 transcript levels significantly decreased in monocytic cells due to estrogen and that the observed decrease in message was blocked by the antagonist fulvestrant. Estrogen reduced CD16 expression and decreased TNF-alpha and IL-1beta release upon CD16 activation but the administration of fulvestrant blocked this decrease. ERalpha was found to interact with a region 5' of the CD16 gene in the presence of estrogen, and site-directed mutational analysis of this region indicated the necessity for an estrogen response element in modulating estrogen effects on CD16 expression. Moreover, both an ERalpha and an ERbeta agonist reduced expression of the CD16 reporter construct suggesting both receptors can play a role in CD16 regulation. In conclusion, CD16 expression can be altered by the activity of ERalpha or ERbeta and our results also show that ERalpha can associate with a region within the CD16 promoter that is important in production of transcript. PMID- 17923259 TI - Interaction between iNOS and COX-2 in hypoxia-induced retinal neovascularization in mice. AB - BACKGROUND: Upregulation of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) has been reported in hypoxia-induced retinal angiogenesis. The aim of our study was to evaluate the possible interaction between the COX and NOS pathways and the effect of this interaction on matrix metalloproteinases 2 (MMP 2) and vascular endothelial growth factor (VEGF) expression and on retinal angiogenesis. METHODS: In this study, the effect of COX-2 or iNOS inhibition on retinal angiogenesis was examined by histopathology. Expression of iNOS, COX-2, MMP-2, and VEGF in the retinas of experimental animals was analyzed using immunohistochemistry, real-time PCR, and Western blotting technologies. RESULTS: Inhibition of COX-2 or iNOS attenuated retinal neovascularization and decreased VEGF and MMP-2 expression. An interaction was found between COX-2 and iNOS expression: the iNOS inhibition decreased COX-2 expression and vice versa. CONCLUSIONS: Our data showed a prominent role of COX-2 and iNOS in hypoxia induced retinal angiogenesis. The interaction between COX-2 and iNOS is likely to produce a cooperative effect on retinal angiogenesis. The changes of MMP-2 and VEGF expression may play a role in the process of retinal neovascularization. PMID- 17923260 TI - Evaluation of aortic remodeling in apolipoprotein E-deficient mice and renovascular hypertensive mice. AB - BACKGROUND: The apolipoprotein E-deficient mouse (ApoE) spontaneously develops hypercholesterolemia and atherosclerotic lesions in large arteries. It is also known that angiotensin II-induced hypertension accelerates the development of atherosclerosis in ApoE mice. The objective of this study was to evaluate the aortic remodeling process in ApoE mice during the early phase of atherosclerosis in two-kidney one-clip hypertensive (2K1C) mice and in mice with the coexistence of atherosclerosis and arterial hypertension. METHODS: Renovascular hypertension was induced in 8- to 9-week-old C57BL/6 (C57) and ApoE and compared to sham animals 28 days later. C57-2K1C and ApoE-2K1C mice showed hypertension, tachycardia, and cardiac hypertrophy of similar magnitude. RESULTS: ApoE and ApoE 2K1C mice showed high levels of plasma cholesterol (4.8- and 3.6-fold) and aorta lipid deposition (85- and 101-fold) compared to C57 mice. The aorta lumen area was increased in C57-2K1C and ApoE-2K1C mice (0.57 +/- 0.04 and 0.55 +/- 0.02 mm(2)) compared to C57 mice (0.50 +/- 0.02 mm(2), p <0.05). The aorta wall area was increased by 20% in C57-2K1C and by 12% in ApoE-2K1C mice compared to C57 and ApoE. CONCLUSIONS: The main finding of this study was the absence of aorta remodeling in ApoE mice at the early stage of atherosclerosis and an outward remodeling of similar magnitude in C57-2K1C and ApoE-2K1C mice. PMID- 17923261 TI - Enhanced glycation of hemoglobin and plasma proteins is associated with increased lipid peroxide levels in non-diabetic hypertensive subjects. AB - BACKGROUND: Accumulating evidences indicate that lipid peroxidation and protein glycation play a vital role in the pathogenesis of cardiovascular disease. The purpose of the present study was to evaluate the levels of lipid peroxides and glycated proteins in non-diabetic hypertensive patients and to assess the possible nexus between them, among these subjects. METHODS: Thirty hypertensive patients and 25 normotensive subjects were enrolled in the present study. Lipid peroxides, glycated hemoglobin, and fructosamine levels were estimated in both groups. RESULTS: Lipid peroxides, glycated hemoglobin, and fructosamine levels were significantly increased in hypertensive subjects in comparison with normotensive subjects. When partial correlation analysis was performed, malondialdehyde was significantly associated with glycated hemoglobin and fructosamine. CONCLUSIONS: An increased glycation of proteins was found in non diabetic hypertensive subjects. These data also support the premise that lipid peroxidation per se plays a role in glycation of hemoglobin and plasma proteins. PMID- 17923262 TI - Improving outcome of human immunodeficiency virus-infected patients in a Mexican intensive care unit. AB - BACKGROUND: In Latin America, insufficient data are available to improve local admission policies for human immunodeficiency virus (HIV) patients in the intensive care units (ICU). We undertook this study to evaluate the outcome and survival determinants of HIV patients in a Mexican ICU during three time periods. METHODS: From December 1985 through January 2006, a clinical chart-based, retrospective study of all HIV patients admitted to the ICU was conducted. Demographic, clinical and laboratory data; disease severity score (APACHE II) and mortality were evaluated. A comprehensive database was created and data were analyzed using survival and regression models. RESULTS: Ninety HIV patients were admitted to the ICU during the study: 16 (18%) in 1985-1992 (non-antiretroviral [ARV]-period), 21 (23%) in 1993-1996 (ARV-period), and 53 (58%) in 1996-2006 (highly active antiretroviral treatment [HAART] period). Leading reasons for admission were the need for mechanical ventilatory support (MVS, 85.5%), septic shock (23%), and non-HIV/AIDS complications (15.5%). Survival in the ICU increased from 12.5% (non-ARV period) to 57% (HAART period). Mortality during ICU stay was associated with MVS (HR: 3.2; 95% CI 1.0-10.2) and APACHE II > or =13 points (HR: 2.2; 95% CI 1.3-4.0). Use of steroids (HR: 0.4; 95% CI 0.2-0.8) and HAART (HR: 0.25; 95% CI 0.1-0.5) were associated with a lower risk of death. In multivariate analysis, septic shock was the main predictor of death in the ICU (HR: 2.4; 95% CI 1.1-5.2) and after discharge. HAART remained as a significant protective factor. CONCLUSIONS: Overall survival in Mexican HIV patients admitted to an ICU has substantially increased in recent years. These data should encourage policies that consider HIV patients as good candidates for receiving intensive care. PMID- 17923263 TI - R219K polymorphism of ATP binding cassette transporter A1 related with low HDL in overweight/obese Thai males. AB - BACKGROUND: ATP binding cassette transporter A1 (ABCA1) plays a role in the initial stage of removing cholesterol from the body via cholesterol efflux. Mutations of this gene cause wide-ranging HDL deficiency, as evident in Tangier disease and familial hypoalphalipoproteinemia. The aim of this study was to elucidate whether the presence of ABCA1 gene polymorphism could be a risk factor for overweight/obesity. METHODS: The presence of R219K and I883M genetic variant was determined by PCR-RFLP analysis in 112 overweight/obese and 117 control subjects of both sexes. Statistical analysis was performed to find an association between polymorphism and lipid data. RESULTS: Overweight/obese men carrying the mutant allele of R219K had lower level of HDL than the control (p = 0.006). However, no positive association was observed using bivariate logistic regression analysis. On the contrary, there was no difference in HDL level among genotypes in I883M polymorphism. Both polymorphisms appeared to be common in Thai ethnic groups. No difference was detected in genotype frequency between the two populations for both polymorphisms. CONCLUSIONS: Although the lower level of HDL in overweight/obese men carrying R219K in comparison to the control suggests the possible involvement of this gene with obesity, further investigations are needed to prove the influence of ABCA1 gene polymorphism on HDL level and to determine whether it could be a genetic determinant of obesity. PMID- 17923264 TI - Prenatal increased asymmetric dimethylarginine is associated with placental heat shock protein 70 and lectin-like oxidized low-density lipoprotein receptor-1 expression. AB - BACKGROUND: Asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, induces endothelial dysfunction by reversibly blocking NO production from L-arginine. To elucidate the association of prenatal status of ADMA with placental heat-shock protein 70 (HSP70) and lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) in normal full-term pregnancies, we evaluated the expression of placental HSP70 and LOX-1. METHODS: Tissue samples of placentas obtained from 60 normal-term pregnancies were categorized into 30 cases with a lower level of prenatal ADMA (0.28 +/- 0.07 microM) and 30 cases with a higher level of prenatal ADMA (1.31 +/- 0.44 microM). We evaluated the placental expression of HSP70 and LOX-1 with Western blot analysis and immunohistochemistry and determined the correlation between HSP70 and LOX-1. RESULTS: We found that prenatal increased ADMA is associated with increased placental HSP70 and LOX-1 expression. CONCLUSIONS: We postulate that higher ADMA levels are associated with excessive levels of oxidative damage and stress markers (HSP, LOX-1) in placental tissues. PMID- 17923265 TI - Alteration of cyclin D1 in Chinese patients with breast carcinoma and its correlation with Ki-67, pRb, and p53. AB - BACKGROUND: For the female population in Asia, systematic investigation on alterations of cyclin D1 in breast carcinoma is rare, and correlation between cyclin D1 expression with clinicopathological parameters, survival rate, and other prognostic marker associated with cell cycle is unclear. METHODS: Expression of cyclin D1 protein, Ki-67, pRb, and p53 was determined by immunohistochemistry in 18 cases of early breast carcinomas and 80 cases of invasive ductal carcinomas. Genetic alteration of cyclin D1 gene and overexpression of cyclin D1 mRNA were detected by Southern blot and RT-PCR, respectively. RESULTS: Expression of cyclin D1 is negative in usual ductal hyperplasia (UDH) and atypical ductal hyperplasia (ADH). However, in 52.0% (51/98) of all breast carcinomas, positive expression of cyclin D1 was observed. Five-year survival rate of the patients with positive expression of cyclin D1 (52.7%) is significantly lower than the cases with negative expression of cyclin D1 (72.1%). Positive rate of cyclin D1 protein in invasive ductal carcinoma (52.5%) is slightly higher than overexpression rate (40.8%) of cyclin D1 mRNA but significantly higher than amplification rate of cyclin D1 gene (18.4%). Expression of cyclin D1 is correlated with Ki-67 expression, but not correlated with pRb and p53 expression. CONCLUSIONS: Positive expression of cyclin D1 could serve as a poor prognostic marker for Chinese patients with breast carcinoma independent of nodal metastasis and clinical stage. Expression of cyclin D1 protein is affected more directly by overexpression of cyclin D1 mRNA rather than cyclin D1 gene amplification. The cooperation between pRb and p53 with cyclin D1 protein in the carcinogenesis of breast carcinoma is not supported by the results. PMID- 17923266 TI - Treatment of seven patients with coarctation of the aorta treated using a Mexican made platinum/iridium stent. AB - BACKGROUND: Nowadays, stenting is the treatment of choice in patients >15 years of age with coarctation of the aorta without hypoplastic aortic isthmus. The platinum/iridium stent manufactured in Mexico may be an affordable alternative with the same benefits as the imported stent. METHODS: This is a series of cases in which we present the immediate results of the first seven patients with coarctation of the aorta treated with the platinum/iridium stent manufactured in Mexico. The first four patients were selected and treated at the Cardiology Hospital of the National Medical Center during the year 2003, two more patients were selected and treated at the National Institute of Cardiology, and another patient was treated at the Military Hospital, Mexico City, in the year 2005. RESULTS: A total of seven patients were successfully treated with the platinum/iridium stent without any complications. Average initial gradient was 56.4 mmHg (range: 30-90 mmHg). In six patients, the final gradient was 0.0 mmHg and in one it was 2 mmHg. In the four patients treated in the Cardiology Hospital of the National Medical Center, mean follow-up time was 17.5 +/- 2.5 months, with a gradient of 0.0 mmHg. All patients are in NYHA Class I. At 12 months follow-up, the gradient remains as 0 mmHg and the patients are still in NYHA Class I functional state. CONCLUSIONS: This stent can be a safe and effective alternative to other stents available on the market for the treatment of coarctation of the aorta. PMID- 17923267 TI - Interleukin-10 gene polymorphism in Parkinson's disease patients. AB - BACKGROUND: The etiology of sporadic Parkinson's disease (PD) is not well established. Recent studies revealed that inflammatory processes might also play an important role in the pathogenesis of PD. We hypothesized that genetically determined differences in the immune response, especially in anti-inflammatory cytokines production, might influence the risk of sporadic PD development and/or onset. To prove this hypothesis, two DNA polymorphisms at IL-10 promoter (-1082 and -519) were examined in sporadic PD patients. METHODS: The study enrolled 341 patients with diagnosed idiopathic PD. All cases of secondary parkinsonism were excluded from the study. For the purpose of this study the patients were also divided into two subgroups: group 1: patients with onset of Parkinson's disease, i.e., <50 years of age (early onset) included 60 patients, as well as group 2: patients with onset of Parkinson's disease >50 years of age (late onset) comprising 281 subjects. Control samples were from 315 randomly selected healthy individuals from the same geographical region who were free from signs of parkinsonism as evaluated by consultant neurologists. PCR-RFLP methods were used for genotyping. RESULTS: No statistically significant differences between PD patients and controls were found in the frequency of a single locus (-1082, -519) of IL-10 promoter. Likewise, haplotype analysis did not demonstrate any significant differences between evaluated groups. The frequency of the evaluated IL-10 genotypes was also similar in EOPD and LOPD patients. CONCLUSIONS: Results from our study revealed that the IL-10 (-1082G>A, -592C>A) polymorphism is not a risk factor of sporadic Parkinson's disease in a Polish population. PMID- 17923268 TI - Effects of smoking and fetal hypokinesia in early pregnancy. AB - BACKGROUND: The aim of this study was to perform qualitative and quantitative ultrasound (US) monitoring of fetal movement in early pregnancy (gestational weeks 10-20) as a component of fetal behavior in women smokers. METHODS: The study included three groups of 20 pregnant women each: non-smokers (group 1), smokers of an average of 10 cigarettes daily (group 2), and smokers of an average of >20 cigarettes daily (chronic smokers; group 3). Two-dimensional US study was performed once during gestational weeks 10-20 by the then standard method of fetal movement monitoring: 1) quantitative measurement of global fetal movements qualitatively verified as brisk or sluggish; 2) quantitative measurement of isolated spontaneous head movements; 3) quantitative measurement of isolated spontaneous arm movements; 4) quantitative measurement of isolated spontaneous leg movements; and 5) M-mode measurement of fetal heart rate. RESULTS: The ratio of brisk to sluggish fetal movements was 82.8% to 17.2%, 79.01% to 20.98%, and 44.25% to 55.75% in groups 1, 2, and 3, respectively (p <0.001). The rate of isolated spontaneous head and arm movements and of the upper cerebral pattern (head and arm movements) was statistically significantly lower in group 3 as compared with groups 1 and 2 (p <0.001), whereas no statistical significance was recorded in isolated spontaneous leg movements (p >0.01). The rate of fetal tachycardia was also significant in group 3, whereas tachyarrhythmia was recorded in seven children born to group 3 mothers (p <0.001). CONCLUSIONS: The present study of the effect of chronic tobacco hypoxia on the components of fetal behavior revealed a positive correlation between global and isolated fetal hypokinesia of the upper cerebral pattern (p <0.001), fetal tachycardia, and tachyarrhythmia in the group of mothers who were chronic smokers (p <0.001). PMID- 17923269 TI - Evaluation of circulating natural type 1 interferon-producing cells in HIV/GBV-C and HIV/HCV coinfected patients: a preliminary study. AB - BACKGROUND: GB virus-C (GBV-C) is a flavivirus that probably influences HIV infection-associated disease among HIV/GBV-C coinfected patients and inhibits the progression of HIV infection to AIDS. To address the possibility of immune modulating effects of GBV-C coinfection in HIV patients, we evaluated interferon producing cells in HIV/GBV-C coinfected patients and compared them to HIV infected patients. METHODS: We performed a pilot study to enumerate interferon producing cell count by two-color flow cytometric analysis and also for determining the frequency of ongoing GBV-C and HCV infection among patients infected with HIV. Then, 83 asymptomatic HIV-positive patients were considered for evaluation of interferon-producing cells. Eighty three patients were stratified in four groups according to the HCV and GBV-C infection status: patients infected with HCV and GBV-C (GBV-C+/HCV+), patients infected with GBV-C but not with HCV (GBV-C+/HCV-), patients infected with HCV but not with GBV-C (GBV-C-/HCV+), and patients not infected by GBV-C and HCV (GBV-C-/HCV-). RESULTS: GBV-C was detected in plasma samples from 15.5% of HIV-infected patients and the frequency of HCV infection was 47.5%. Interferon-producing cells in GBV-C coinfected individuals revealed a wider range of numbers; however, there was no significant difference in the interferon-producing cell count among HIV-infected individuals. CONCLUSIONS: It does not appear that GBV-C coinfection affects generation or redistribution of interferon-producing cells in HIV-infected patients with relatively intact immune system. PMID- 17923270 TI - Exercise duration rather than peak oxygen uptake better correlates with Fev1 and inspiratory capacity in chronic obstructive pulmonary disease. AB - BACKGROUND: Exercise duration of constant-load endurance tests has been recently demonstrated to be more sensitive in detecting the changes after bronchodilator administration than either walking distance or peak oxygen uptake in patients with COPD. In the present study, we questioned whether exercise duration of progressive load is better correlated with forced expiratory volume in 1 sec and inspiratory capacity than other indices of submaximal exercise test during stable period in patients with COPD. METHODS: Thirty-three stable COPD patients, and 26 age- and BMI-matched control subjects were recruited. Resting pulmonary function tests and incremental cycle exercise tests were performed. All indices of exercise and pulmonary function tests including exercise duration, peak oxygen uptake (VO(2peak)), and forced expiratory volume in 1 sec (FEV(1)) were recorded. RESULTS: Exercise capacity was lower in COPD than age- and BMI-matched controls as shown by exercise duration and ergometric work (535 +/- 159 vs. 705 +/- 115 sec, p <0.001 and 89.2 +/- 26.6 vs. 117.8 +/- 19.5 W, p <0.001). Statistical analysis revealed that exercise duration slightly better correlates with FEV(1) and inspiration capacity (IC) (R = 0.600, p <0.001; R = 0.615, p <0.001) than peak oxygen uptake (R = 0.284, p >0.05; R = 0.127, p >0.05) in stable period COPD patients. CONCLUSIONS: There is an impairment of aerobic capacity in stable period COPD patients compared to healthy controls. Exercise in COPD patients is ventilation-limited and exercise duration with respect to peak VO(2) is better correlated with FEV(1) and IC in stable COPD patients undergoing progressive-load cycling exercise. PMID- 17923271 TI - Prevalence of overweight/obesity in Chinese children. AB - BACKGROUND: The purpose of this paper was to report the prevalence of overweight/obesity by area, age, and gender in the year 2000 and to explore at what age adiposity rebound as measured by BMI occurs for Chinese children. METHODS: A cross-sectional study was carried out among 262,738 children aged 3.5 6.4 years in 26 counties/cities in China. Weight and height were measured from March 2000 through July 2000 by local trained health workers. International age- and gender-specific BMI cut-off points were used to obtain comparable prevalence rates of overweight/obesity. Multiple logistic regression was used to estimate odds ratios (OR) of being overweight/obesity for different comparisons. RESULTS: There were 19,390 overweight (16,738) or obese (2,652) children, resulting in an overall prevalence rate of 7.4% (95% CI: 7.3-7.5%). Girls were more likely to be overweight or obese than boys (OR = 1.14, 95% CI: 1.11-1.17). Compared with children living in the southern urban area, multivariate ORs for those living in northern rural and southern rural areas were 2.58 and 1.15, respectively. Compared with children aged 6.0-6.4 years old, multivariate ORs for children aged 3.5, 4.0, 4.5, 5.0 and 5.5 years were 1.96, 1.88, 1.56, 1.53, and 1.11, respectively (p<0.001). CONCLUSIONS: Childhood overweight/obesity in Chinese children in 2000 is similar to the conditions of Great Britain or the U.S. in the 1980s or earlier. Age at adiposity rebound as measured by BMI for Chinese children is >6.5 years old. PMID- 17923272 TI - Lifestyle-related determinants and serum adiponectin concentrations in a general population of Japanese females. AB - BACKGROUND: Serum adiponectin concentration plays a role in the development of metabolic and atherosclerotic diseases. Although elucidating the determinants that affect circulating adiponectin concentrations including lifestyle factors are important, little is known about the relationships between lifestyle and adiponectin in the general Japanese female population. METHODS: A total of 311 Japanese females (mean age 54 years) with normal lifestyles and taking no medications were enrolled in a community-based cross-sectional study to investigate which lifestyle factors were associated with serum adiponectin concentrations. Adiponectin was measured with an enzyme-linked immunosorbent assay. The modified Breslow's index (a well-known comprehensive index) was used in assessing lifestyle factors. RESULTS: The group of moderate drinkers (consuming 1-3 go/day on average) had significantly higher serum adiponectin concentrations than that of nondrinkers (consuming <1 go/day) (15.0 +/- 7.2 vs. 12.0 +/- 6.5 microg/mL). Using a partial correlation analysis on serum adiponectin concentrations adjusted for age and all lifestyle factors, there were significant positive correlations with age (r = 0.197) and alcohol intake (r = 0.130) and a significant inverse correlation with body mass index (BMI) (r = 0.178). CONCLUSIONS: Among many lifestyle factors, in addition to maintaining decreased BMI levels, moderate alcohol intake habits may have a significant, independent, and positive effect on adiponectin concentrations in the general population of Japanese females, similar to that of Caucasian populations. PMID- 17923273 TI - Circulating leptin and osteoprotegerin levels affect insulin resistance in healthy premenopausal obese women. AB - We investigated the relationship between circulating leptin and osteoprotegerin (OPG) levels and insulin resistance assessed by HOMA-IR in premenopausal obese and normal weight women. Thirty four obese women (age 31 +/- 8 years) (BMI 35 +/- 4 kg/m(2)) with 19 healthy controls (age 31 +/- 7 years) (BMI <25 kg/m(2)) (BMI 21 +/- 2 kg/m(2)) were included in the study. Women were healthy and had no osteoporosis. Circulating leptin levels were significantly higher in obese women (17.11 +/- 2.05 ng/mL vs. 8.38 +/- 4.71 ng/mL, p <0.0001) and decreased OPG levels were found (14.7 +/- 7.15 pg/mL vs. 19.17 +/- 6.37 pg/mL, p = 0.03). Leptin showed a positive correlation with BMI (r = 0.851, p <0.0001), waist-to hip ratio (r = 0.692, p <0.0001), fasting insulin (r = 0.441, p <0.001), HOMA-IR (r = 0.412, p = 0.002), fibrinogen (r = 0.387, p = 0.004), uric acid (r = 0.293, p = 0.033), hematocrit (r = 0.394, p = 0.003), systolic (r = 0.504, p <0.0001), and diastolic blood pressure (r = 0.363, p = 0.008). OPG showed a negative correlation with insulin (r = -0.341, p = 0.013) and HOMA-IR (r = -0.324, p = 0.018). In obese women group, the regression equation of HOMA-IR was (HOMA-IR = [0.095 x leptin]-[0.051 x OPG] + 1.71). However, there was no relation between leptin and OPG levels. In conclusion, circulating leptin and OPG levels were related to insulin resistance in premenopausal obese women. However, leptin had no interference in OPG in premenopausal women. PMID- 17923274 TI - Unusual finding of HTLV-I infection among Amazonian Amerindians. AB - Human T-cell lymphotropic virus type II is a retrovirus endemic in Amerindian communities throughout the American continent, although some Amerindian groups that apparently emerged from the same ethnic root as HTLV-II carriers do not secrete antibodies against the virus and show very low prevalence for human T cell lymphotropic virus type I. In this study, sera from 487 Amazonian amerinds were screened for HTLV type I and II antibody by the gelatin particle agglutination assay and ELISA and confirmed by Western blot and indirect immunofluorescence assay. None was positive for HTLV type II. One young healthy male of Waiapi ethnicity was reactive with HTLV-I and was confirmed by Western blot assay and indirect immunofluorescence test. The absence of HTLV type II infection among these Amerindian communities would suggest a behavior pattern distinct from other groups in the American continent. Also, the very low prevalence of HTLV type I infection among these ethnic groups probably indicates contamination by blood transfusion (external transmission route). PMID- 17923275 TI - Extremely prolonged INR associated with warfarin in combination with both trazodone and omega-3 fatty acids. AB - We report the case of a 65-year-old Caucasian male who had been taking warfarin for 6 months after an episode of postoperative pulmonary thromboembolism (PTE). His medical history was otherwise insignificant. He received a prescription of trazodone and fish oil for not-very-well defined complaints. Two weeks after taking these medications, when he routinely checked his international normalized ratio (INR), he arrived at the Emergency Department (ED) with a test result showing an INR of 8.06. He was admitted for observation and all medications were discontinued. His INR returned to normal within 2 days. He was then restarted on his previous warfarin dose, while other drugs were not restarted. Two weeks later, his coagulation profile was within the desired therapeutic range. Coadministration of warfarin with omega-3 fatty acids can lead to additional anticoagulation. This can result from changes either in platelet aggregation or vitamin K-dependent coagulation factors. Trazodone also has interactions with warfarin through not well-understood mechanisms. Although drug interaction reference texts classify warfarin-trazodone interaction as late-onset and clinically insignificant, this has been questioned in other studies. This particular case illustrates a possible interaction between warfarin and these two medications. PMID- 17923278 TI - Induction of the NAD(P)H:quinone oxidoreductase 1 by ketoconazole and itraconazole: a mechanism of cancer chemoprotection. AB - Studies in many carcinogen-induced animal models and cell lines demonstrated that azole antifungal drugs are therapeutically effective against different types of cancer. Yet, the molecular mechanisms involved are still poorly understood. Therefore, we examined the ability of three structurally different antifungal drugs, ketoconazole (KTZ), itraconazole (ITZ), and fluconazole (FLZ) to induce the expression of NAD(P)H:quinone oxidoreductase 1 (Nqo1), an enzyme known to play an important role in xenobiotic and carcinogen detoxifications. We showed that KTZ and ITZ, but not FLZ, induced Nqo1 mRNA and enzymatic activity levels in a concentration- and time-dependent manner in wild-type but not aryl hydrocarbon receptor (AhR)-deficient Hepa 1c1c7 cells. Furthermore, KTZ and ITZ increased Nqo1 de novo RNA synthesis without significantly affecting the levels of existing RNA, suggesting a transcriptional mechanism is involved. This study provides the first evidence for the ability of KTZ and ITZ to induce the Nqo1 gene expression through an AhR-dependent mechanism. PMID- 17923276 TI - Relationship between lipoprotein(a) and thyroid function status in the general population. PMID- 17923279 TI - Proanthocyanidin from grape seeds inactivates the PI3-kinase/PKB pathway and induces apoptosis in a colon cancer cell line. AB - The aim of this investigation was to evaluate the chemopreventative/antiproliferative potential of a grape seed proanthocyanidin extract (GSPE) against colon cancer cells (CaCo2 cells) and to investigate its mechanism of action. GSPE (10-100 microg/ml) significantly inhibited cell viability and increased apoptosis in CaCo2 cells, but did not alter viability in the normal colon cell line (NCM460). The increased apoptosis observed in GSPE treated CaCo2 cells correlated with an attenuation of PI3-kinase (p110 and p85 subunits) and decreased PKB Ser(473) phosphorylation. GSPE might thus exert its beneficial effects by means of increased apoptosis and suppression of the important PI3-kinase survival-related pathway. PMID- 17923280 TI - Translational control: a target for cancer therapy. AB - Cap-mediated translation is the default mechanism for the synthesis of proteins in eukaryotic cells. Increasingly, malignant cells have been found to have deregulation of this process. Return of normal translational control is associated with loss of tumorigenic potential in pre-clinical models. Currently, a variety of novel therapeutics are in development targeting this mechanism as a treatment for cancer. PMID- 17923281 TI - The honor of protecting participants while conducting research or clinical projects. PMID- 17923283 TI - Top 10 lists - medications associated with adverse events and medications involved with errors. PMID- 17923284 TI - Nursing roles in health care decision making. PMID- 17923285 TI - Assisted living nursing practice: the language of dementia: theories and interventions. AB - The person with dementia uses behavior to communicate, but their behavior is altered by the combination of neurological damage and impairment, altered interpersonal relationships and reactions of others, and the individual's loss or weakening of their lifelong defenses or coping mechanisms. This article discusses the routes by which behavior can be understood and describes a constellation of needs of a person with dementia that has a unique fit with person-centered care. Three evidence-based models (theories) and interventions specific to dementia behaviors are discussed: the Need-Driven Dementia-Compromised Behavior Model, the Progressively Lowered Stress Threshold Model, and the utilization of self identity roles. Montessori-based activities are another approach to person centered dementia care that respect, as do the models, the dignity, worthiness and interests of the person afflicted with dementia. The models discussed in this article all seek to improve the quality of life of the person with dementia. Other than those at the profound end stage of dementia, most sufferers can communicate feelings. Subjective quality of life must be determined based on the self-report of the person suffering with dementia so that treatment interventions and effectiveness are grounded in that person's reality. PMID- 17923286 TI - Integrating geriatric resources into the classroom: a virtual tour example. AB - As the older adult population increases, nursing students at all levels need geriatric content and access to geriatric resources. The Virtual Tour (VT) assignment, a combination of Web-based geriatric resources and applied learning activities, provides a simple way to integrate Web-based resources into classroom learning. VTs provide students a guide or "road map" to practical Web-based resources for client care. Evaluation data support that students like VTs and gain useful information for practice. VTs provide an easy way to expand geriatric resources available to students and to complement classroom content. PMID- 17923287 TI - Comprehensive fall prevention programs across settings: a review of the literature. AB - The prevention and management of falls across health care and community settings continues to be one of the greatest challenges in geriatric nursing. This article reviews the literature on fall prevention and management and provides information on national programs and resources, as well as public policy related to falls in the elderly. PMID- 17923288 TI - A student-led demonstration project on fall prevention in a long-term care facility. AB - Falls are a frequent and serious problem facing people aged 65 and older. The incidence of falls increases with greater numbers of intrinsic and extrinsic risk factors and can be reduced by risk modification and targeted interventions. Falls account for 70% of accidental deaths in persons aged 75 and older. Mortality due to falls is significantly higher for older adults living in extended care facilities versus those living in the community. Our objective was to evaluate the effectiveness of a fall prevention training program in a long-term care setting. A single-group repeated-measure design was used, guided by the Precede Proceed framework. A comprehensive review of the literature and a concept analysis guided the development of testing and educational materials for all nursing and ancillary facility staff. Preliminary testing provided baseline data on knowledge related to fall prevention. Pre- and posttests, a fall prevention newsletter, and informational brochures were distributed to nursing staff and ancillary personnel at training sessions. Certified nursing assistant (CNA) champions were identified and given peer leadership training. "Quick Tips" fall prevention badges were also distributed to staff. Graduate students led interdisciplinary environmental rounds weekly, and new falls were reviewed on a daily basis by the interdisciplinary team. A 60-day posttest evaluated retention of fall prevention knowledge. Fall rates at baseline and for 2 months after the intervention were compared. Preliminary survey data revealed fall prevention learning opportunities, with a pretest mean score of 86.78%. Qualitative data were coded and revealed specific learning gaps in intrinsic, extrinsic, and organizational causes of falls. The 60-day posttest mean score was 90.69%; a paired t test (t score = -1.050; P = .057) suggested that learning may have taken place; however, differences in scores did not reach statistical significance. The fall rate before training was 16.1%; 30-day posttraining fall rate was 12.3%, and 60-day postintervention fall rate was 9%. Based on the program results, the model was expanded from long-term care to the university hospital system and outpatient clinics in the same community. The collaboration between a school of nursing and 1 long-term care facility led to the adoption of a significant quality improvement program that was subsequently extended to a local hospital and clinic system. Student-led projects designed to teach community service learning can be meaningful and can lead to changes in patient safety and quality of care. PMID- 17923290 TI - Estimation of SVE closure time. AB - The ability to predict the time when to stop a SVE system is critical when designing and operating a SVE remediation system. Completed research on the tailing performance of SVE, allowed the development of the closure time index (CTI) concept. CTI is based on breakthrough curves for both lab-scale experiments and a field-scale application of SVE. Application of the CTI concept allows estimation of the time when the SVE system could be shutdown. Shutting down a SVE system at the appropriate time minimizes operational time and reduces clean-up costs as demonstrated by the field case data that was tested. PMID- 17923289 TI - In their own words: older male prisoners' health beliefs and concerns for the future. AB - U.S. prisons are experiencing an exponential growth in inmates aged 50 years and older, a group with disproportionately high disease burden. The purpose of this study was to examine, in largely exploratory terms, the health beliefs and concerns of older male inmates and the health challenges they anticipate facing upon their return to the community. Results indicate that there is much to be gained from the assessments and insights of older prisoners with regard to health changes that occur during incarceration, health programs that they desire, the reasons for their confidence (or lack thereof) in health self-management, and fears about their health upon release. Geriatric nurses are well positioned to heed these important insights of inmates and translate them into steps for 1) preventing many of the health deteriorations experienced by older prisoners and 2) advocating for more seamless health care when incarcerated offenders transition back into the community. PMID- 17923291 TI - Bioremediation of anthracene contaminated soil in bio-slurry phase reactor operated in periodic discontinuous batch mode. AB - Bioremediation of soil-bound anthracene was studied in a series of bio-slurry phase reactors operated in periodic discontinuous/sequencing batch mode under anoxic-aerobic-anoxic microenvironment using native soil microflora. Five reactors were operated for a total cycle period of 144 h (6 days) at soil loading rate of 16.66 kg soil/m(3)/day at 30 +/- 2 degrees C temperature. The performance of the bioreactors was studied at various substrate loading rates (volumetric substrate loading rate (SLR), 0.1, 0.2 and 0.3g anthracene/kg soil/day) with and without bioaugmentation (domestic sewage inoculum; 2 x 10(6) CFU/g of soil). Control reactor (without microflora) showed negligible degradation of anthracene due to the absence of biological activity. The performance of the bio-slurry system with respect to anthracene degradation was found to depend on both substrate loading rate and bioaugmentation. Application of bioaugmentation showed positive influence on the rate of degradation of anthracene. Anthracene degradation data was analysed using different kinetic models to understand the mechanism of bioremediation process in the bio-slurry phase system. Variation in pH/oxidation-reduction potential (ORP), soil microflora and oxygen consumption rate correlated well with the substrate degradation pattern observed during soil slurry phase anthracene degradation. PMID- 17923292 TI - Sustainable development of process facilities: state-of-the-art review of pollution prevention frameworks. AB - Pollution prevention (P2) strategy is receiving significant attention in industries all over the world, over end-of-pipe pollution control and management strategy. This paper is a review of the existing pollution prevention frameworks. The reviewed frameworks contributed significantly to bring the P2 approach into practice and gradually improved it towards a sustainable solution; nevertheless, some objectives are yet to be achieved. In this context, the paper has proposed a P2 framework 'IP2M' addressing the limitations for systematic implementation of the P2 program in industries at design as well as retrofit stages. The main features of the proposed framework are that, firstly, it has integrated cradle-to gate life cycle assessment (LCA) tool with other adequate P2 opportunity analysis tools in P2 opportunity analysis phase and secondly, it has re-used the risk based cradle-to-gate LCA during the environmental evaluation of different P2 options. Furthermore, in multi-objective optimization phase, it simultaneously considers the P2 options with available end-of-pipe control options in order to select the sustainable environmental management option. PMID- 17923293 TI - Injuries sustained to the upper extremity due to modern warfare and the evolution of care. AB - The formation of the American Society for Surgery of the Hand was related to world conflicts and hostilities. Therefore, it is appropriate that upper extremity surgeons understand injuries resulting from modern-day combat. Because of ongoing warfare, many countries have experienced a large increase in the number of wounded service members and civilians, particularly wounds of the extremities. As a result of increased rate of survival in battlefield trauma in part because of the use of modern body armor, there is increasing complexity of extremity injuries that require complex reconstructions. Decreased mortality and a consequent increase in the incidence of injured extremities underline the need for the development of new treatment options. The purpose of this presentation is to describe upper-extremity injury patterns in modern warfare, the levels of care available, and the treatment at each level of care based on the experience of the United States Military Medical Support System. PMID- 17923294 TI - Long-term follow-up evaluation of bilateral total hand loss. AB - PURPOSE: Bilateral total hand loss is rare, and the challenges faced by the patient immediately after the loss are quite different from the long-term responses to the physical loss of both hands. This study evaluates the problems of these patients years after their injuries. The goal is to evaluate long-term adaptation to the loss and the degree of successful coping achieved by the patients. METHODS: Seven subjects in this descriptive study participated in an interview and physical exam. They were asked about their physical, psychological, and social problems. Functional Independence Measure (FIM) and Functional Assessment Measure (FAM) were used to assess the levels of the major functional areas. RESULTS: All of the subjects considered their handicap a serious disfigurement but not an impediment to finding a marriage partner or pursuing educational goals. All of the subjects, however, were dependent economically although they wanted to be employed based on their capabilities. Although 2 subjects used prostheses for activities of daily living, they all needed assistance for self-care. Three of the subjects had diagnosed psychological problems of anxiety and depression. CONCLUSIONS: Bilateral total hand loss does not simply represent double the problems of unilateral hand loss. The problems resulting from the loss of both hands are physical, social, psychological, and economic and unique to these patients. The strength of the subjects came from having good family relations, stable environments, and family emotional support to help solve the numerous problems encountered on a daily basis. The subjects are able to maintain stable personal lives because they experience respect and comprehension of their problems by family members. PMID- 17923295 TI - Comparing the outcome of a carpal tunnel decompression at 2 weeks and 6 months. AB - PURPOSE: The purpose of the study was to evaluate if 2-week Levine score can provide an adequately responsive outcome measure in carpal tunnel decompression by comparing it with 6-month score. METHODS: The treatment outcome of 300 patients with carpal tunnel decompression was determined by using Levine score at 2 weeks and 6 months after surgery. The mean age of patients was 56 years, 71% (214) were women, and 55% (167) of operations were performed on the right hand. All patients were scored using the Levine questionnaire preoperatively and at 2 weeks and 6 months from date of surgery. The correlation between the scores was evaluated. RESULTS: Although statistical significance was found between the preoperative score and the scores at both 2 weeks and 6 months, no statistical difference was found between the scores at 2 weeks and 6 months after surgery. Multiple regression analysis with the 2 week-6 month score difference as the dependent variable shows a predictable outcome at 2 weeks. CONCLUSIONS: We conclude that the Levine score at 2 weeks is a reliable, responsive, and practical instrument for outcome measure in carpal tunnel surgery. It coincides with suture removal and provides a convenient and predictive assessment of the medium-term results in a high percentage of treated patients. We conclude that the 2 time points, 2 weeks and 6 months, are equivalent in outcome because a less than 10-point difference is not clinically meaningful. PMID- 17923296 TI - Outcomes of carpal tunnel surgery with and without supervised postoperative therapy. AB - PURPOSE: To assess if a formal 2-week hand therapy improves outcomes and justifies its expense. METHODS: A prospective randomized study was completed using a contemporary short incision and a 2-week program of therapy. Patients were randomized into 2 treatment groups: one group received instruction on home therapy exercises to be followed after carpal tunnel release, and a second group received the home program in addition to a therapist-directed program for 2 weeks. Variables measured were patient age, gender, preoperative and postoperative pain scores, grip and pinch strengths, return to modified and regular work, insurance coverage, and job category. Both groups were followed for 6 months postoperatively. RESULTS: One hundred fifty patients (110 women and 40 men) entered and completed the study. The average age was 46 years (range, 29-70 years). The average age, gender distribution, insurance coverage, and breakdown of job categories between groups was not statistically significant. There was no difference in return to work times between those with and without postoperative therapy; however, patients covered by workers' compensation insurance were slower to return to both modified and regular work compared with the other groups. The postoperative grip and pinch strengths, pain and Disabilities of the Arm, Shoulder, and Hand scores did not show statistical differences between groups at any of the measured time periods. Depending on insurance carrier, directed therapy added $600 to $900 to the cost of care. CONCLUSIONS: The current randomized study failed to show benefit in a 2-week course of hand therapy after carpal tunnel release using a short incision. The cost of supervised therapy for an uncomplicated carpal tunnel release seems unjustified. PMID- 17923298 TI - Clinical course of the non-operated hand in patients with bilateral idiopathic carpal tunnel syndrome. AB - PURPOSE: Patients with bilateral carpal tunnel syndrome (CTS) who had unilateral carpal tunnel surgery usually ask about the prognosis for the non-operated, opposite hand. In this study, we investigated the effects of unilateral carpal tunnel surgery on the clinical course of the non-operated, opposite hand in bilateral idiopathic CTS. METHODS: In this prospective study, only patients who had bilateral idiopathic CTS but had unilateral carpal tunnel surgery were included. Sixty-six patients were included in the study. All patients were women with the mean age 47 years and mean duration of symptoms 4 years. Forty-six surgeries were performed on right hands, and 20 surgeries performed on left hands. Forty-eight (73%) patients were housewives. Their non-operated, opposite hands were evaluated before and 6 months after unilateral surgery. The evaluations consisted of subjective symptoms and objective findings. Paired t test and chi-square test were used for statistical analysis. RESULTS: There were no statistically significant differences between the means of symptom severity score, functional status score, power and pinch grip strength before and 6 months after unilateral surgery in the non-operated, opposite hands. There was no statistically significant differences among the percentages of the Tinel's sign, Phalen test, and electrophysiological changes before and 6 months after unilateral surgery on the non-operated, opposite hands. During the study period, 57 (86%) patients had surgery or intended to have surgery for their opposite hands. CONCLUSIONS: It seems that unilateral surgery for bilateral idiopathic CTS does not affect the clinical course of the non-operated, opposite hand. We recommend that each hand of patients with bilateral CTS be managed separately, based on their symptoms. PMID- 17923299 TI - Endoscopic decompression of the ulnar nerve at the elbow. AB - PURPOSE: The ideal operative treatment for cubital tunnel syndrome, the second most common form of peripheral compression neuropathy, remains controversial. We therefore reviewed our series of endoscopically assisted ulnar nerve decompression at the elbow to determine the effectiveness of the procedure, which was intended to minimize perioperative morbidity and scar discomfort. METHODS: In 36 patients (ages 22-76 years) with clinical McGowan grade I (4 patients), II (21 patients), and III (11 patients) and electrophysiologic signs of cubital tunnel syndrome (35 primary, 1 recurrent), 20 cm of the ulnar nerve was released through a 3.5-cm-long skin incision above the medial epicondyle. A 4-mm, 30 degrees standard endoscope and custom-made guiding-dissecting tool were utilized during the procedure, and the mean postoperative follow-up examination was 14 months (range 6-19). RESULTS: No macroscopically visible nerves and vessels were injured during the procedure. The only postoperative complication was hematoma in one patient that resolved after conservative management. One case was converted from endoscopic to open because of a ganglion that surrounded the nerve in the forearm. There was no scar discomfort (ie, painful neuroma, impaired sensibility, or burning sensation) or elbow extension deficit after surgery, and surgical wounds all healed within a week. Outcomes were excellent in 21 of 36 cases and good in 12 of 36 cases. All patients improved electrophysiologically after surgery, were satisfied with the procedure, returned to full activities within 3 weeks, and would have the procedure again. CONCLUSIONS: By using a safe and reliable endoscopic technique characterized by a short incision, minimum soft tissue dissection, and early postoperative mobilization, we were able to preserve the benefits of conventional approaches (namely, complete release and good visualization), while avoiding problems such as painful scarring and elbow contracture. PMID- 17923300 TI - The radial nerve in the brachium: an anatomic study in human cadavers. AB - PURPOSE: To explore the course of the radial nerve in the brachium and to identify practical anatomic landmarks that can be used to avoid iatrogenic injury during humerus fracture fixation. METHODS: Data were collected from 27 adult cadaveric specimens, including 18 embalmed cadavers and 9 fresh-frozen limbs. Measurements were taken using osseous landmarks to define the relationship of the radial nerve and the posterior and lateral humerus. The extremities were studied further to determine the association of the radial nerve and anatomic landmarks on both longitudinal and cross-sectioned specimens. RESULTS: A 6.3 cm +/- 1.7 segment of radial nerve was found to be in direct contact with the posterior humerus from 17.1 cm +/- 1.6 to 10.9 cm +/- 1.5 proximal to the central aspect of the lateral epicondyle, centered within 0.1 cm +/- 0.2 of the level of the most distal aspect of the deltoid tuberosity. The radial nerve lay in direct contact with the periosteum in all specimens, without evidence of a structural groove in the humerus in any specimen. On entering the anterior compartment, the radial nerve had very little mobility as it was interposed between the obliquely oriented lateral intermuscular septum and the lateral aspect of the humerus. As it extended distally, the nerve coursed anterior to the humerus and became protected by brachialis muscle at the level of the proximal aspect of the lateral metaphyseal flare. CONCLUSIONS: The radial nerve is at risk of injury with fractures of the humerus and with subsequent operative fixation in 2 areas. The first is along the posterior midshaft region for a distance of 6.3 cm +/- 1.7 centered at the distal aspect of the deltoid tuberosity. The second is along the lateral aspect of the humerus in its distal third from 10.9 cm +/- 1.5 proximal to the lateral epicondyle to the level of the proximal aspect of the metaphyseal flare. The deltoid tuberosity is a consistent and practical anatomic landmark that can be used to determine the level of the radial nerve along the posterior aspect of the humerus during operative fixation from an anterior approach. PMID- 17923301 TI - Motor reinnervation of a denervated muscle by using a sensory nerve: an experimental study on gluteus maximus muscle of the rat. AB - PURPOSE: This study aimed at developing a new muscle reinnervation technique using a sensory nerve. METHODS: We attempted innervation of the rat gluteus maximus muscle using the lateral femoral cutaneous nerve (LFCN). We placed the gluteus maximus muscle into the fibroadipose tissue in the distribution of the LFCN in 24 rats. In one group, the original innervation remained intact. In the second and third groups, the muscles were denervated, and in the third group, the proximal end of the nerve to the gluteus maximus was sutured to the distal end of the divided LFCN. We compared muscle reinnervations of the groups by using electrophysiologic evaluation of the muscle contractions, light microscope evaluation of the axonal regenerations, and scanning electron microscope evaluation of the actin-myosin structures of the muscles at the end of an elapsed waiting period. RESULTS: At the end of electrophysiologic evaluation, the mean area of compound muscle action potentials measured in group 1 was 3.8 ms/mV; in group 2, 0.0; and in group 3 (experimental group), 0.5. Axonal regeneration was observed distal to the coaptation, and actin-myosin structures were mostly spared in group 3. CONCLUSIONS: This study explored the feasibility of a new flap prefabrication method that aims at developing reinnervation of a denervated muscle by means of a sensory nerve. In light of histologic and electrophysiologic findings, this type of reinnervation is possible. PMID- 17923302 TI - Biomechanical influence of the vincula tendinum on digital motion after isolated flexor tendon injury: a cadaveric study. AB - PURPOSE: The vincula are specialized mesotendinous structures attaching to the flexor tendons of the hand. In addition to providing vascular supply to the tendons, the vincula can be mechanically important. The purpose of this study was to quantify the influence of intact vincula on digital flexion after flexor tendon laceration and to assess the ultimate strength and stiffness of the vincula. METHODS: The index, middle, and ring fingers of 12 fresh-frozen cadaveric fingers were dissected free at the level of the metacarpophalangeal joint, preserving at least 10 cm of the flexor and extensor tendons. A 9.8-N load was applied to each flexor tendon, and using digital photography and image analysis software, the degree of flexion at the proximal and distal interphalangeal joints and excursion of tendons proximal to the metacarpophalangeal joint was recorded before and after division of the flexor digitorum profundus and flexor digitorum superficialis tendons at their insertions. Load to failure and stiffness of the vincula were measured via a uniaxial material testing apparatus. Analysis of means was performed with a paired t-test. RESULTS: After division of the flexor digitorum superficialis tendon, proximal interphalangeal joint flexion secondary to the influence of the intact vincula was 93% of that compared with the uninjured digit. Distal interphalangeal joint flexion after flexor digitorum profundus transection was 69% of normal. The increased excursion of transected tendons compared with testing before division was 4 mm for flexor digitorum superficialis and 2 mm for flexor digitorum profundus. Load to failure was 27 N, and stiffness was 6 N/mm. CONCLUSIONS: The vincula breve can facilitate digital flexion after distal tendon transection, allowing tendons to act indirectly across the interphalangeal joints. The intact vincula breve can facilitate an almost normal range of motion across the interphalangeal joints, making the diagnosis of a flexor tendon injury difficult. In the immediate postinjury period, the vincula breve can hold a divided tendon within a few millimeters of its insertion. Testing against resistance is important to avoid missing the diagnosis of a tendon injury. PMID- 17923303 TI - Spontaneous flexor tendon rupture of the flexor digitorum profundus secondary to an anatomic variant. AB - We report a case of the flexor digitorum profundus tendon rupture of the little finger, which was predisposed by an anatomic variation of the tendon. Intraoperative findings and magnetic resonance imaging of the opposite hand suggested that the flexor digitorum profundus tendons of the ring and the little finger bifurcated. The patient had tendon reconstruction and regained function. We believe that reconstructing the tendon so that it resembles the normal anatomy prevents the recurrence of tendon rupture. PMID- 17923304 TI - Elbow dislocation with intra-articular fracture: the results of operative treatment without repair of the medial collateral ligament. AB - PURPOSE: To determine the effectiveness of a protocol for the treatment of fracture-dislocations of the elbow based on the concept that, if dislocation of the elbow with associated fractures can be made to resemble a simple elbow dislocation by repairing or reconstructing the fractured structures, repair of the medial collateral ligament (MCL) will not be necessary. METHODS: Over a 5 year period, a single surgeon operated on 34 patients with a posterior dislocation of the elbow associated with one or more intra-articular fractures. The mean age of these 19 men and 15 women was 48 years. Associated fractures included the capitellum, trochlea, and lateral epicondyle in 3 patients; the olecranon in 1 patient; and the radial head in 30 patients (with concomitant fracture of the coronoid process-the so-called "terrible triad" of the elbow-in 22 patients, and concomitant fracture of the coronoid and olecranon in 1 patient). Operative treatment consisted of open reduction internal fixation (ORIF) or prosthetic replacement of all fractures and reattachment of the origin of the lateral collateral ligament (LCL) complex to the lateral epicondyle. The MCL was not repaired. RESULTS: Two patients (1 with a terrible triad injury and 1 with fracture of the capitellum and trochlea) had postoperative instability related to noncompliance, had reconstructive procedures, and were considered failures. An average of 32 months after injury, the remaining 32 patients regained an average of 120 degrees ulnohumeral motion and 142 degrees forearm rotation. Twenty-five of 34 patients (74%) had good or excellent results according to the system of Broberg and Morrey. Patients with terrible triad injuries had an average of 117 degrees ulnohumeral motion and 137 degrees forearm rotation, and 17 of 22 patients (77%) had good or excellent results. CONCLUSIONS: MCL repair is unnecessary in the treatment of dislocation of the elbow with associated intra-articular fractures, provided that the articular fractures and the LCL are repaired or reconstructed. PMID- 17923305 TI - The effect of medial collateral ligament repair tension on elbow joint kinematics and stability. AB - PURPOSE: Medial collateral ligament (MCL) repair is commonly performed for the management of acute or subacute instability after elbow dislocations and fracture dislocations. The effectiveness of transosseous repair of the MCL, as is typically performed clinically, in restoring the normal kinematics and stability of the elbow is of interest as is the effect of MCL tensioning on the initial stability of the elbow. The purpose of this study was to determine whether suture repair of the MCL is able to restore the normal kinematics and stability of the elbow and to determine the optimal initial MCL repair tension. METHODS: Six cadaveric upper extremities were mounted in an upper limb joint simulator. Simulated active and passive elbow flexion was generated while the kinematics were measured with the arm in the dependent and the valgus gravity-loaded orientations. After testing the intact elbow, the MCL was released at its humeral attachment and repaired using a transosseous suture technique at three different repair tensions: 20, 40, and 60 N. RESULTS: Medial collateral ligament repair using a transosseous suture technique restored the kinematics and stability of the MCL-deficient elbow. Motion pathways were affected by the magnitude of initial MCL tension. For all arm orientations and forearm positions, the 20-N and 40-N repairs were not statistically different from each other or from the intact MCL. The 60-N repairs, however, were often statistically different than the other groups, suggesting an overtightening that tended to pull the ulna into a varus position-especially in the midrange of flexion. CONCLUSIONS: These data suggest that MCL repair using transosseous sutures provide adequate joint stability to permit early motion. There is a broad range of acceptable tensions for MCL repair, which is a favorable, clinically relevant finding. Clinical studies are needed to validate these in vitro results. PMID- 17923306 TI - The effect of the annular ligament on kinematics of the radial head. AB - PURPOSE: There is evidence that the radial head translates during pronation and supination. This study measured radial head movement and the location of the pronation-supination axis of the forearm with and without the annular ligament. METHODS: Thirteen cadaveric arms were attached to a custom-built frame. Pronation and supination were achieved by actuation of the pronator teres and biceps tendons, respectively. Motion was captured by video cameras via marker arrays attached to the distal radius, distal ulna, humerus, and radial head. Three pronation-supination trials were performed with the annular ligament intact and the elbow positioned at 90 degrees . The sequence was repeated after transection of the annular ligament. RESULTS: The radial head traveled an average of 2.1 mm in the anteroposterior direction and 1.6 mm in the mediolateral direction during forearm rotation. After annular ligament resection, travel of the radial head increased by an average of 44% in the mediolateral direction and by 24% in the anteroposterior direction. On average, the mean pronation-supination axis (screw displacement axis) passed 1.4 mm +/- 1.9 medial to the center of the capitellum and through the center of the ulnar head. Loss of the annular ligament did not change the location of the pronation-supination axis. CONCLUSIONS: During forearm rotation, the radial head travels to a greater extent in the anteroposterior direction than in the mediolateral direction. Loss of the stabilizing effect of the annular ligament increases the travel mediolaterally more than anteroposteriorly. The pronation-supination axis of the forearm is nearly constant and is not affected by annular ligament transection. PMID- 17923307 TI - The distal biceps tendon: footprint and relevant clinical anatomy. AB - PURPOSE: There is little information in the literature describing the anatomy of the biceps tendon insertion. The purpose of this study was to map the footprint of the biceps tendon insertion on the bicipital tuberosity and to report on the relevant anatomy to assist surgeons with correct tendon orientation during surgical repair. METHODS: Fifteen fresh-frozen adult upper extremities were used in this study. The relationships between the long head of the biceps tendon, the short head of the biceps tendon, the muscle bellies, and the distal tendon orientation were examined. The length, width, and area of the biceps tendon insertion were measured. RESULTS: In all specimens examined, the biceps musculotendinous unit rotated 90 degrees externally from origin to insertion. The long head of the distal tendon was inserted onto the proximal aspect of the bicipital tuberosity, while the short head of the distal tendon was inserted onto the distal aspect of the tuberosity. The lacertus fibrosus, in all specimens, originated from the distal short head of the biceps tendon. On average, the biceps tendon insertion started 23 mm distal to the articular margin of the radial head. The average length of the biceps tendon insertion on the tuberosity was 21 mm, and the average width was 7 mm. The average total area of the biceps tendon insertion (footprint) was 108 mm(2). The average area of the long head of the biceps tendon insertion was 48 mm(2), and the average area of the short head of the biceps tendon insertion was 60 mm(2). CONCLUSIONS: Landmarks have been identified that will allow anatomic orientation of the distal biceps tendon during operative repair. The distal short head of the biceps tendon has a consistent relationship with the lacertus fibrosus and a distinct insertion on the bicipital tuberosity. The dimensions of the distal biceps tendon footprint have been determined to assist with bone tunnel or suture anchor placement during surgical repair. PMID- 17923308 TI - Radioulnar heterotopic ossification after distal biceps tendon repair: results following surgical resection. AB - PURPOSE: The purpose of this study is to evaluate the clinical outcome of patients who had excision of a radioulnar heterotopic ossification (HO) as a complication of a distal biceps tendon repair. The hypothesis is that there are no measurable clinical losses that persist after excision. METHODS: Eight consecutive patients were identified between 1996 and 2005. All were treated with HO excision using a standard surgical technique and rehabilitation protocol. These individuals were studied and compared to a matched cohort of 8 patients who had a distal biceps tendon repair with a similar surgical technique that was uncomplicated. All study patients were evaluated at a minimum 1-year follow-up with physical examination, isokinetic dynamometry, and outcome measures. Comparisons were made both between groups as well as side-to-side within groups. RESULTS: At follow-up examination, the mean arc of forearm rotation in the HO group measured 151 degrees. The mean arc of forearm rotation in the control group measured 165 degrees. With the numbers available, no measurable differences in arc of motion were identified between groups (p > .05). When compared to the normal, uninvolved side, patients who developed HO lost an average of 9 degrees of forearm pronation (p < .01). No differences were identified between the HO and control groups with respect to isokinetic torque, endurance strength, or Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire (American Academy of Orthopaedic Surgeons, Rosemont, IL) scores (p > .05). CONCLUSIONS: When patients develop motion-limiting HO after distal biceps tendon repair, surgical resection can lead to a functional recovery of elbow and forearm motion. Biceps strength can be maintained with no measurable differences in clinical outcome when compared to individuals who do not suffer this complication following distal biceps repair. PMID- 17923309 TI - Brachialis muscle tear mimicking an intramuscular tumor: a report of two cases. AB - Isolated brachialis muscle tears are rare. The rarity causes the misdiagnosis of this injury. We report 2 cases of males with an isolated brachialis muscle tear who were referred to our hospital with a diagnosis of a neoplasm in the arm. Both of the patients improved completely without radical treatment. PMID- 17923311 TI - Palmar bypass for digital ischemia. AB - Although vein bypass grafting in lower extremity and heart tissue ischemia is well documented, little information exists with regard to its selective use for digital ischemia in the hand. Resection and ligation and interposition grafting are most commonly performed to treat digital ischemia and have received the most attention in the literature. We report on 3 cases of successful vein bypass grafting from the distal forearm to the palmar arch or common digital artery as a technique to reverse current and prevent further digital tip ischemia or necrosis. PMID- 17923310 TI - Treatment of proximal interphalangeal dorsal fracture-dislocation injuries with dynamic external fixation: a pins and rubber band system. AB - PURPOSE: Unstable, dorsal, intra-articular, fracture-dislocations of the proximal interphalangeal (PIP) joint can be difficult to treat and often lead to long-term pain, stiffness, and functional deficit. We present the outcomes of patients sustaining such injuries that were treated by a novel dynamic external fixator. This fixator uses a system of K-wires and rubber bands that maintains a concentrically reduced PIP joint while allowing for early motion. METHODS: Fourteen patients with unstable, dorsal fracture-dislocation injuries of the PIP joint were treated between September 2001 and January 2006. Eight were available for follow-up evaluation at an average of 26 months. We measured PIP range of motion and grip strength, and assessed pain on a visual analog scale. Demographic information about the original injury was recorded. New radiographs were obtained to assess joint congruency and the presence of arthritis or articular step-off deformity. RESULTS: In the 8 patients available for follow-up evaluation, the average motion of the affected PIP joint was from 1 degrees (range 0 degrees to 5 degrees) to 89 degrees (range 75 degrees to 110 degrees). Grip strength was 92% (range 71% to 110%) of the unaffected hand. The average score on the visual analog pain scale was 0.6 (range 0-1.5). There were few complications. Radiographs at follow-up evaluation showed a concentric reduction in all joints, but with evidence of a small step-off deformity or arthritis in 5 patients. CONCLUSIONS: The dynamic external fixator studied is an effective method of treating unstable, dorsal fracture-dislocation injuries. Outcomes compared favorably with those of other similar devices studied in the literature. PMID- 17923312 TI - Closed degloving of the thumb. AB - We present a work-related injury where the drums of a printing machine caused a closed degloving of the distal phalanx of the thumb, which represents detachment of the soft tissue from the bone. PMID- 17923313 TI - Distal ulnar implant arthroplasty as a definitive treatment of a recurrent giant cell tumor. AB - Giant-cell tumors of the distal ulna are frequently recurrent and difficult to treat. A Darrach-type resection can lead to functional impairments of the wrist and hand. A case of a 42-year-old man, who underwent a distal ulna arthroplasty for recurrent giant-cell tumor of the distal ulna, is presented. The patient had two previous operations in an attempt to curette the cavity, either alone or filled with allograft bone chips. En bloc resection and distal ulnar implant arthroplasty provided a good functional outcome without any evidence of tumor recurrence at a 2-year follow-up evaluation. PMID- 17923314 TI - Can brief interventions by hand surgeons influence medical students toward a career in hand surgery? AB - PURPOSE: In 2004, the American Society for Surgery of the Hand and the American Association for Hand Surgery formed a joint task force to deal with the difficulty of promoting hand surgery as a career choice to medical students. The purpose of this study was to use a previously validated format to expose medical students to a brief intervention early in their medical education that might influence their perceptions of hand surgery as a career choice. METHODS: First year medical students were asked to rank 11 items on a Likert scale from 1 (not important) to 5 (very important) regarding their beliefs about hand surgery as a career before and after a brief 1-hour presentation by a hand surgeon. Additional information about career choice and prior exposure to hand surgery was obtained before the intervention, and student interest in hand surgery was measured after the intervention. RESULTS: Of 110 first-year medical students, 72 (65%) students completed both the pre-lecture survey and the post-lecture survey. Of the 72 students with matched responses, 8 (11%) ranked surgery as their first career choice, whereas 13 (18%) ranked a surgical subspecialty as their first choice. Thirty-nine (54%) students stated that the lecture raised their interest in hand surgery as a possible career, and 36 (50%) were interested in doing an elective in it while in medical school. Nonparametric tests showed improvement in knowledge after the lecture for length of training, technical aspects, intellectual challenge, and fellowship requirements. CONCLUSIONS: A positive information-providing encounter with a hand surgeon may favorably influence the perceptions of first-year medical students toward hand surgery. PMID- 17923315 TI - Lateral epicondylitis: review and current concepts. AB - Lateral epicondylitis is a painful and functionally limiting entity affecting the upper extremity and is frequently treated by hand surgeons. The anatomic basis of the injury to the extensor carpi radialis brevis origin appears to be multifaceted, involving hypovascular zones, eccentric tendon stresses, and a microscopic degenerative response. Although many treatments have been advocated, there is little clear consensus on which modality works best, for both conservative and operative options. In this article, we present an overview of this difficult problem and an evidence-based review of treatment choices. PMID- 17923316 TI - The role of neurodiagnostic studies in nerve injuries and other orthopedic disorders. AB - Neurodiagnostic techniques, such as nerve conduction studies, needle electromyography studies, intraoperative nerve monitoring, and evoked potentials provide useful information for practicing orthopedic surgeons to help localize central from peripheral nervous system lesions. For peripheral nerve lesions, it helps localize the level of the nerve dysfunction (for example, root versus plexus versus peripheral nerve). These techniques are well established and routinely used. Newer techniques have emerged that aim to simplify the technical process of performing these studies and help reduce the discomfort associated with these studies. Many of these newer techniques, however, are in their infancy, and their role in routine use for neurodiagnostic purposes is not clear. This review examines the various types of nerve injuries commonly encountered in orthopedic surgery practice and the role of electrodiagnostic (neurodiagnostic) techniques in diagnosing these conditions. PMID- 17923317 TI - A method of defining palpable landmarks for the ligament-splitting dorsal wrist capsulotomy. AB - PURPOSE: The concept of a ligament-splitting dorsal capsulotomy of the wrist has been defined and has been shown to have practical applications. Due to the abundance of peritendinous tissue, however, the ligaments are often difficult to visualize. This article presents a method of defining incision capsular lines based on reliably palpable landmarks. METHODS: The palpable landmarks include the sulcus between the scaphoid and trapezoid, the dorsal tubercle of the triquetrum, and the midpoint between Lister's tubercle and the dorsal rim of the sigmoid notch. These points identify the bisection lines of the dorsal intercarpal and dorsal radiocarpal ligaments. A radial-based capsulotomy can be easily elevated by incising the dorsal wrist joint capsule using these landmarks and then extending the incision along the dorsal rim of the distal radius to the radial styloid process. RESULTS: This method of defining capsular incision lines based on palpable landmarks was used on 253 consecutive dorsal wrist arthrotomies with excellent exposure, accurate splitting of the dorsal radiocarpal and intercarpal ligaments, and no complications. CONCLUSIONS: Using specific, palpable landmarks on the dorsal wrist, an accurate estimation of the locations and courses of the dorsal radiocarpal and intercarpal ligaments can be reliably made. Even when poorly visualized, these ligaments can be split longitudinally in a reliable fashion to create a standard, ligament-sparing dorsal capsulotomy. PMID- 17923318 TI - Intraoperative nerve stimulation: a simple, effective, and inexpensive alternative to standard devices. AB - Many surgical procedures require intraoperative stimulation of peripheral nerves. Using a pair of conventional bipolar forceps and an anesthetic impulse generator (Stimlocator, Model SL3, B. Braun Australia Pty Ltd, Australia), we have developed a simple, effective, and inexpensive alternative to standard nerve stimulation devices that enables the accurate localization of motor fascicles intraoperatively. Bipolar forceps provide better control than a monopolar electrode, as the current generated across the nerve fibers generates an action potential that is then propagated down the axon. This inexpensive and reusable device is routinely used at Royal Children's Hospital, Melbourne, and its efficacy and ease of use has been demonstrated over a long period. PMID- 17923321 TI - Intraosseous blood supply to the distal humerus. PMID- 17923319 TI - Volar plating for intra-articular fracture of the base of the proximal phalanx. AB - Comminuted fractures involving the articular surface of the base of the proximal phalanx are relatively rare and pose a challenging problem for hand surgeons because of the difficulty in achieving an accurate reduction and secure fixation of the articular surface. These fractures usually comprise a volar base fracture associated with a central depression of the articular surface. We describe a technique for open reduction and plate fixation of intra-articular fractures of the base of the proximal phalanx through a volar A1 pulley approach. Compared with the dorsal approach, this technique offers the advantages of direct visualization of the volar base fragment and the depressed central fragment, allowing for a more accurate reduction and rigid internal fixation using a volar buttress plate. In addition, there is no interference with extensor apparatus. Although our experience is limited to 4 patients, we have had a positive experience with this technique. The technique is useful for internal fixation of intra-articular fractures of the base of the proximal phalanx. PMID- 17923322 TI - Butyrylcholinesterase, ApoE and Alzheimer's disease in a population from the Canary Islands (Spain). AB - AIM: Cholinergic dysfunction is a major neurochemical feature in Alzheimer's disease (AD), accountable for many cognitive dysfunctions and some psychiatric symptoms. Butyrylcholinesterase (BChE) is one of the cholinesterases with increased activity in the brain of Alzheimer's patients. Several mutations code for different BChE, such as the K variant, which is the most common and is capable of reducing BChE activity by 30%. We studied the relationship between this K variant and Alzheimer's disease in our population from the Canary Islands (Spain). PATIENTS AND METHODS: We used DNA PCR-RFLP techniques to compare 282 patients who had been diagnosed with probable Alzheimer's disease--according to NINCS-ADRDA criteria--with 312 control subjects confirmed to be free of cognitive impairment as assessed by using the CAMDEX cognitive subscale CAMCOG. RESULTS: In our population the K variant of BChE is linked to the age of diagnosis of Alzheimer's disease, since AD individuals with this allele presented the disease at a later stage. No other susceptibility relations are exposed in this study. In addition, the BChE allelic frequencies in our population are higher than those previously reported. PMID- 17923323 TI - Phosphorylation of c-Jun N-terminal kinase isoforms and their different roles in spinal cord dorsal horn and primary somatosensory cortex. AB - The present study was undertaken to investigate whether isoforms of c-Jun N terminal kinase (JNK 46 kDa and 54 kDa), one component of the mitogen-activated protein kinase (MAPK) family, might show region-related differential activation patterns in both naive and pain-experiencing rats. In naive rats, no significant difference was observed in total expression level of the two JNK isoforms between spinal cord and primary somatosensory cortex (S1 area). However, phosphorylated JNK 46 kDa was normally expressed in the S1 area, but not in the spinal cord, while neither of the two structures contained phosphorylated JNK 54 kDa. Subcutaneous bee venom (BV)-induced persistent pain stimulation resulted in a significant increase in the phosphorylation of both JNK isoforms in each area for a long period (lasting at least 48 h). Nevertheless, JNK 46 kDa exhibited a much higher activation than JNK 54 kDa in the spinal cord, whereas the same noxious stimulation elicited evident activation of JNK 54 kDa in the S1 area, leaving JNK 46 kDa less affected. Intraplantar injection of sterile saline solution, causing acute and transient pain, produced almost the same changes in activation profile of the two JNK isoforms as found in the BV-treated rats. These results implicate that individual members of the JNK family may be associated with specific regions of nociceptive processing. Also, the two JNK isoforms are supposed to function differently according to their locations within the rat central nervous system. PMID- 17923324 TI - Dyslexia and attentional shifting. AB - Dyslexia is a neurocognitive deficit primarily expressed in reading difficulties, but also affecting non-linguistic performance. Several studies report that dyslexics perform differently in the attentional blink paradigm, which indicates an impaired capacity to rapidly shift visual attention. However, attentional shifting can occur at different levels of cognitive processing, and it is unclear whether dyslexic attentional shifting is impaired at all levels, or only at the peripheral levels. We studied performance on a task-switching paradigm by dyslexics and normal readers to test whether the difficulty with attentional shifting occurs at the level of central cognitive processing. We found no specific impairments in task-switching in dyslexics. However, dyslexics performed generally much more slowly across all conditions than normal readers. We conclude that while dyslexics have a problem with attentional switching at a perceptual level, their capacity to rapidly switch between tasks is normal. Our findings add to previous studies indicating that dyslexic problems with shifting visual attention are caused by anomalies in more peripheral neural pathways, such as the magnocellular layers in the lateral geniculate nucleus. PMID- 17923325 TI - Study on olfactory function in GABAC receptor/channel rho1 subunit knockout mice. AB - The GABAC receptor/channel rho1 subunit plays an important role in the inhibitory pathway and sensory processing in the retina and spinal cord. Although it was suggested that the rho1 subunit plays a role in olfactory sensations, the precise role of the rho1 subunit in olfactory sensory function is still not clear. In the present study, we report that olfactory function was significantly altered in rho1 subunit knockout (rho1-/-) mice compared to its wildtype counterpart. The rho1 subunit mRNA, detected by reverse transcription (RT)-PCR experiments, was expressed in the olfactory bulb of wild-type mice. Expression of rho1 subunit proteins in the olfactory bulb was detected by immunohistochemistry in mitral cells in the mitral cell layer. Neither mRNA nor proteins of the rho1 subunit were found in olfactory bulb neurons in rho1-/- mice. Alterations of olfactory function in rho1-/- mutant mice compared to their wildtype littermates were examined by olfactory behavioral test. We found that sensitivity to the smell of citral odorant in rho1-/- mice was significantly greater compared to that of wildtype mice. Our results indicate that the GABAC rho1 subunit acts in olfactory bulb neurons as an inhibitory modulator that affects the process of olfactory signaling transmission. PMID- 17923326 TI - P-glycoprotein has negligible effects on estradiol and testosterone in mice. AB - P-glycoprotein (P-gp) is an important factor at the blood-brain barrier preventing passage of a wide variety of substances into the brain. Several studies have provided evidence that some drugs and certain steroid hormones are substrates or inhibitors of P-gp. However, the situation is unclear with regard to gonadal steroids, which have considerable central nervous effects. In vitro, experiments on the relationship between estradiol and P-gp are equivocal. We used abcb1ab knock-out mice and wild-type mice to determine the uptake of [3H]-17-beta estradiol and [3H]-testosterone into the cerebrum and other organs after subcutaneous administration. The organ/plasma quotients showed no significant group effects. We concluded that P-gp does not influence the penetration of testosterone and estradiol to a biologically significant extent. PMID- 17923327 TI - Different roles of spinal p38 and c-Jun N-terminal kinase pathways in bee venom induced multiple pain-related behaviors. AB - Our previous studies have established the idea that different types of pain induced by subcutaneous bee venom (BV) injection might be mediated by different spinal signaling pathways. To further testify this hypothesis, the present investigation was designed to detect whether spinal p38 and c-Jun N-terminal kinase (JNK) pathways are equally or differentially involved in the development of persistent spontaneous nociception (PSN), primary heat and mechanical hyperalgesia, and mirror-image heat (MIH) hypersensitivity in the BV model, by evaluating the effects of intrathecal (i.t.) pre-administration of a p38 inhibitor SB239063 and a JNK inhibitor SP600125 in the conscious rat. The results showed that i.t. pre-treatment with either SB239063 or SP600125 caused a significant prevention of BV-induced persistent paw flinching reflex in a dose related manner, with the former exhibiting much stronger inhibition than the latter. Moreover, the same doses of SB239063 and SP600125 also exhibited different suppressive actions on the induction of primary heat hyperalgesia and MIH hypersensitivity. That is, SP600125 produced a larger increase of thermal latency than SB239063 in the injected paw, whereas SB239063 mainly affected the value measured in the non-injected paw. Pre-treatment with neither SB239063 nor SP600125 had any effect on BV-evoked mechanical hyperalgesia. Taken together, these data suggest that activation of p38 in the spinal cord preferentially contributes to the development of PSN and MIH hypersensitivity under pathological state, while spinal JNK signaling pathways might play more important roles in inducing primary heat hyperalgesia. PMID- 17923328 TI - Effects of intra-articular ketamine on pain and somatosensory function in temporomandibular joint arthralgia patients. AB - Recent studies have hypothesized that peripheral glutamate receptors could be implicated in deep craniofacial pain conditions. In this study 18 temporomandibular joint (TMJ) arthralgia patients received intra-articular injections of the N-methyl-D-aspartate (NMDA) receptor antagonist, ketamine, or normal saline to study in a cross-over, double-blinded, placebo-controlled manner the effect on TMJ pain and somatosensory function. Spontaneous pain and pain on jaw function was scored by patients on 0-10 cm visual analogue scale (VAS) for up to 24h. Quantitative sensory tests (QST): tactile, pin-prick, pressure pain threshold and pressure pain tolerance were used for assessment of somatosensory function at baseline and up to 15 min after injections. There were no significant effects of intra-articular ketamine over time on spontaneous VAS pain measures (ANOVA: P=0.532), pain on jaw opening (ANOVA: P=0.384), or any of the somatosensory measures (ANOVA: P>0.188). The poor effect of ketamine could be due to involvement of non-NMDA receptors in the pain mechanism and/or ongoing pain and central sensitization independent of peripheral nociceptive input. In conclusion, there appears to be no rationale to use intra-articular ketamine injections in TMJ arthralgia patients, and peripheral NMDA receptors may play a minor role in the pathophysiology of this disorder. PMID- 17923329 TI - A randomized, controlled trial of emotional disclosure in rheumatoid arthritis: can clinician assistance enhance the effects? AB - Emotional disclosure by writing or talking about stressful life experiences improves health status in non-clinical populations, but its success in clinical populations, particularly rheumatoid arthritis (RA), has been mixed. In this randomized, controlled trial, we attempted to increase the efficacy of emotional disclosure by having a trained clinician help patients emotionally disclose and process stressful experiences. We randomized 98 adults with RA to one of four conditions: (a) private verbal emotional disclosure; (b) clinician-assisted verbal emotional disclosure; (c) arthritis information control (all of which engaged in four, 30-min laboratory sessions); or (d) no-treatment, standard care only control group. Outcome measures (pain, disability, affect, stress) were assessed at baseline, 2 months following treatment (2-month follow-up), and at 5 month, and 15-month follow-ups. A manipulation check demonstrated that, as expected, both types of emotional disclosure led to immediate (post-session) increases in negative affect compared with arthritis information. Outcome analyses at all three follow-ups revealed no clear pattern of effects for either clinician-assisted or private emotional disclosure compared with the two control groups. There were some benefits in terms of a reduction in pain behavior with private disclosure vs. clinician-assisted disclosure at the 2-month follow-up, but no other significant between group differences. We conclude that verbal emotional disclosure about stressful experiences, whether conducted privately or assisted by a clinician, has little or no benefit for people with RA. PMID- 17923330 TI - Confirmation of the efficacy of a novel formulation of metaflumizone plus amitraz for the treatment and control of fleas and ticks on dogs. AB - A novel spot-on formulation containing metaflumizone plus amitraz (ProMeris/ProMeris Duo for Dogs, Fort Dodge Animal Health, Overland Park, KS) was evaluated in four laboratory studies to confirm efficacy against fleas and ticks on dogs for 1 month. Three different strains of cat flea (Ctenocephalides felis felis) and four tick species were used. Rhipicephalus sanguineus and Dermacentor variabilis were evaluated concurrently in two studies and Ixodes scapularis and Amblyomma americanum in one study each. In all studies, dogs were randomly allocated to treatment groups and compared with nontreated dogs. One study also included a placebo treatment and a commercial product containing fipronil plus S methoprene. All treatments were applied to the skin at a single spot between the scapulae on Day 0. Dogs were infested with fleas and/or ticks prior to treatment and then reinfested at weekly intervals for 6 weeks after treatment and evaluated for efficacy at 1 or 2 days after treatment and each reinfestation. These studies confirmed that treatment with ProMeris for Dogs at the proposed commercial dose rate rapidly controlled existing infestations of fleas and ticks on dogs. Treatment provided control of reinfesting fleas for up to 6 weeks and at least 4 weeks control of ticks. Efficacy was confirmed in a variety of dog breeds against three different flea strains and four common species of ticks found on dogs in the United States. PMID- 17923331 TI - Efficacy of a novel formulation of metaflumizone plus amitraz for the treatment of demodectic mange in dogs. AB - A novel spot-on formulation containing metaflumizone plus amitraz (ProMeris/ProMeris Duo for Dogs, Fort Dodge Animal Health, Overland Park, KS) was evaluated for efficacy against demodectic mange mites in naturally infested dogs. Sixteen dogs were allocated to two equal groups and individually housed. Eight of the dogs were treated topically with metaflumizone plus amitraz at the proposed minimum dose rate (20mg/kg of each of metaflumizone and amitraz, 0.133ml/kg) on Days 0, 28, and 56. The other eight were treated with metaflumizone plus amitraz at the proposed minimum dose rate on Days 0, 14, 28, 42, 56, and 70. Mite numbers were estimated from skin scrapings taken on Days -3 to -1, 28, 56, and 84. Clinical signs of mange and the extent of demodectic lesions on each dog were evaluated when skin scrapings were conducted. Efficacy of the treatment was based on a reduction in mite numbers and an assessment of the clinical signs associated with canine demodectic mange. Treatment at monthly or two-weekly intervals for 3 months resulted in a rapid reduction in mite numbers (>94 and >99% for the monthly and two-weekly treatments, respectively) and an improvement in clinical signs. Success rates, based on zero mite counts in skin scrapings at Day 84 were 42.9 and 62.5% of dogs for the monthly and two-weekly regimens, respectively. PMID- 17923332 TI - Showing that you care: the evolution of health altruism. AB - Human behavior regarding medicine seems strange; assumptions and models that seem workable in other areas seem less so in medicine. Perhaps, we need to rethink the basics. Toward this end, I have collected many puzzling stylized facts about behavior regarding medicine, and have sought a small number of simple assumptions which might together account for as many puzzles as possible. The puzzles I consider include a willingness to provide more medical than other assistance to associates, a desire to be seen as so providing, support for nation, firm, or family provided medical care, placebo benefits of medicine, a small average health value of additional medical spending relative to other health influences, more interest in public that private signals of medical quality, medical spending as an individual necessity but national luxury, a strong stress-mediated health status correlation, and support for regulating health behaviors of the low status. These phenomena seem widespread across time and cultures. I can explain these puzzles moderately well by assuming that humans evolved deep medical habits long ago in an environment where people gained higher status by having more allies, honestly cared about those who remained allies, were unsure who would remain allies, wanted to seem reliable allies, inferred such reliability in part based on who helped who with health crises, tended to suffer more crises requiring non-health investments when having fewer allies, and invested more in cementing allies in good times in order to rely more on them in hard times. These ancient habits would induce modern humans to treat medical care as a way to show that you care. Medical care provided by our allies would reassure us of their concern, and allies would want you and other allies to see that they had pay enough to distinguish themselves from posers who didn't care as much as they. Private information about medical quality is mostly irrelevant to this signaling process. If people with fewer allies are less likely to remain our allies, and if we care about them mainly assuming they remain our allies, then we want them to invest more in health than they would choose for themselves. This tempts us to regulate their health behaviors. This analysis suggests that the future will continue to see robust desires for health behavior regulation and for communal medical care and spending increases as a fraction of income, all regardless of the health effects of these choices. PMID- 17923333 TI - Gastroenterology. Preface. PMID- 17923334 TI - Chronic gastrointestinal symptoms in the elderly. AB - Typical and atypical symptoms from acid reflux, dyspepsia, chronic constipation, fecal incontinence, and irritable bowel syndrome are extremely common in adults and remain so in the geriatric population. The presence of these problems may have profound effects on the functional status, independence, and quality of life in the vulnerable older population, making it essential for physicians to inquire actively about them and to be able to recognize atypical presentations when appropriate. This article summarizes the definitions, epidemiology, clinical presentation, and impact of these common problems in the geriatric patient. PMID- 17923335 TI - The anorexia of aging. AB - Undernutrition is common in the elderly, particularly those in nursing homes and other institutions. It is associated with substantial adverse effects. The age associated physiologic reduction in appetite and food intake, which has been termed "the anorexia of aging," contributes to the development of pathologic anorexia and undernutrition. This article reviews age-related changes to appetite, food intake, and body composition; undernutrition in the elderly; and the factors contributing to physiologic and pathologic anorexia and undernutrition. PMID- 17923336 TI - The aging gut: physiology. AB - Changes in the physiology of the gastrointestinal tract with aging are less obvious than are seen in other organs, such as the brain. Nevertheless, physiologic changes play a role in the anorexia of aging, postprandial hypotension, aspiration pneumonia, increased Clostridium difficile infections, fecal incontinence, gallstones, and altered drug metabolism. PMID- 17923337 TI - Gastrointestinal bleeding in older adults. AB - Gastrointestinal bleeding in older adults is a frequent cause of hospital admissions. The presence of multiple comorbidity conditions and greater medication use in this age group influence the clinical outcome. The selection of diagnostic and therapeutic maneuvers often depends more on local expertise and availability. This article addresses clinical presentation, etiology, diagnosis, and treatment of upper and lower gastrointestinal bleeding in elderly individuals. PMID- 17923338 TI - Gastric emptying, diabetes, and aging. AB - Gastric emptying is mildly slowed in healthy aging, although generally remains within the normal range for young people. The significance of this is unclear, but may potentially influence the absorption of certain drugs, especially when a rapid effect is desired. Type 2 diabetes is common in the elderly, but there is little data regarding its natural history, prognosis, and management. This article focuses on the interactions between gastric emptying and diabetes, how each is influenced by the process of aging, and the implications for patient management. PMID- 17923339 TI - Inflammatory bowel diseases in the elderly. AB - Inflammatory bowel disease (IBD) in elderly individuals is associated with a unique set of challenges, some of which are related to age. This article examines the diagnosis and management of IBD in the context of recent advances in the understanding of its pathogenesis, and newer therapeutic modalities that have been possible from these advances. PMID- 17923340 TI - Constipation and irritable bowel syndrome in the elderly. AB - Constipation and irritable bowel syndrome are very common in older persons. Constipation is over diagnosed and over treated in older persons. Lifestyle changes, osmotic laxatives, and lubiprostone are the approaches of choice for the management of constipation. Irritable bowel syndrome should be considered in the differential diagnosis of abdominal pain, diarrhea and constipation in older persons. PMID- 17923341 TI - Diarrheal diseases in the elderly. AB - Diarrhea in the elderly population is one disease that needs special attention in treatment and management, especially in acute- and long-term care residents, because of their multiple comorbidities, immunosenescence, frailty, and poor nutritional status. Close follow-up to ensure adequate hydration and electrolyte replacement and infection control measures to contain outbreaks should be emphasized to caregivers and nursing staff in acute- and long-term care facilities. Although C difficile colitis causes significant morbidity and mortality in this population, judicious use of antibiotics is important to decrease the incidence and recurrence of the disease. When the diarrhea is chronic and all stool testings and serologies have been performed, the patient may benefit from endoscopy and colonoscopy for biopsy. Attentive and vigilant nursing staff is crucial in the timely diagnosis and treatment of diarrheal diseases to improve quality of life and reduce mortality. PMID- 17923342 TI - Fecal incontinence in older adults. AB - Fecal incontinence is an underreported and underappreciated problem in older adults. Although fecal incontinence is more common in women than in men, this difference narrows with aging. Risk factors that lead to the development of fecal incontinence include dementia, physical disability, and fecal impaction. Treatment options include medical or conservative therapy for older adults who have mild incontinence, and surgical options can be explored in selected older adults if surgical expertise is available. PMID- 17923343 TI - Mesenteric ischemia in the elderly. AB - Intestinal ischemia is a relatively common disorder in the elderly and, if not treated promptly, still carries a high morbidity and mortality rate. High degree of clinical suspicion is of paramount importance in diagnosis, because there is no specific laboratory test available and physical examination findings may be subtle. Once the diagnosis is made, management relies on early resuscitation, identification, and treatment of the predisposing conditions, along with careful planning of the therapeutic invasive interventions, which altogether may help reduce the mortality and morbidity associated with this condition. PMID- 17923344 TI - Aging liver and hepatitis. AB - Chronic liver disease and cirrhosis are the tenth leading causes of death in the United States and results in approximately 25,000 deaths annually. As life expectancy in developed countries has increased, so has the number of elderly patients who have liver disease. With an aging population and chronic liver disease becoming an increasingly significant cause of morbidity and mortality, the various causes for hepatitis will need to be evaluated and available treatments considered, even in elderly population. Common causes for hepatitis in elderly individuals include viral, autoimmune, and drug-induced hepatitis, but evidence for treatment of this population is limited. This article reviews the likely causes of hepatitis in elderly individuals and discusses evidence for treating this population. PMID- 17923345 TI - Alcoholic liver disease in the elderly. AB - Although per capita alcohol consumption, and thus the prevalence of alcoholic liver disease, decreases generally with age in Europe and in the United States, recently an increase in alcohol consumption has been reported in individuals over 65 years. Reasons explaining this observation may include an increase in life expectancy or a loss of life partners and, thus, loneliness and depression. Although ethanol metabolism and ethanol distribution change with age, and an elderly person's liver is more susceptible to the toxic effect of ethanol, the spectrum of alcoholic liver diseases and their symptoms and signs is similar to that seen in patients of all ages. However, prognosis of alcoholic liver disease in the elderly is poor. In addition, chronic alcohol consumption may enhance drug associated liver disease and may also act as a cofactor in other liver diseases, such as viral hepatitis and nonalcoholic fatty liver disease. PMID- 17923346 TI - Mandibular radiomorphometric measurements as indicators of possible osteoporosis in postmenopausal women. AB - OBJECTIVE: The use of mandibular anatomic indicators on panoramic radiographs, i.e. the number of lost teeth, mandibular cortical width at the mental region (MCW), panoramic mandibular index (PMI), alveolar crest resorption degree (M/M ratio) and morphologic classification of the mandibular inferior cortex (MIC grade) can be useful in the evaluation of bone resorption in different age groups of women to determine the presence of osteoporosis. The purpose of this study was to assess the validity of mandibular radiomorphometric measurements and to determine the frequency of tooth loss in postmenopausal women. SUBJECTS AND METHODS: An assessment of the number of lost teeth, MCW, PMI, M/M ratio and MIC grade was performed on dental panoramic radiographs in a group of 133 postmenopausal women 38-80 years-of-age. BMD at the lumbar spine was measured by dual energy X-ray absorptiometry. BMD values were categorized as normal (T-score greater than 1.0), and as indicative of osteopenia (T-score -1.0 to -2.5) or osteoporosis (T-score less than -2.5) according to the World Health Organization classification. RESULTS: In our study when the T-score at the lumbar spine is decreased, the age of menopause is increased, and the MCW is decreased to a point of statistical significance. A decrease in MCW by 1mm increases the likelihood of osteopenia or osteoporosis to 43%, having taken into consideration the effect of the years elapsed since menopause. It was also shown that age, years since menopause, MCW value, and the number of teeth lost have a statistically important effect on the incidence of moderate or severe cortical erosion. Moreover, when the MCI is C2 or C3 (mild or severe erosions) the age is increased, the years since menopause are increased and the MCW is decreased to a point of statistical significance. As far as tooth loss is concerned, an increase by 1 unit in the number of teeth lost, increase the likelihood of moderate or severe erosion to 6%, having taken into account the years elapsed since menopause. Our study also demonstrated that postmenopausal women tend to lose their teeth at an age older than 50 years. They usually lose the 1st and 2nd mandibular molars and the 1st and 2nd maxillary premolars. Loss of front teeth and canines occurs at an age older than 60 years (except for the lateral maxillary incisors). At a younger age they tend to lose the 2nd maxillary premolars more frequently than their mandibular counterparts. CONCLUSIONS: In conclusion, panoramic radiographs constitute an integral part of almost every routine dental evaluation and can be useful for the early diagnosis of osteoporosis in postmenopausal women. Dentists have sufficient clinical and radiographic information that enables them to play a significant role in patient screening for osteoporosis. PMID- 17923347 TI - Microemulsions containing lecithin and sugar-based surfactants: nanoparticle templates for delivery of proteins and peptides. AB - Two pseudo-ternary systems comprising isopropyl myristate, soybean lecithin, water, ethanol and either decyl glucoside (DG) or capryl-caprylyl glucoside (CCG) as surfactant were investigated for their potential to form microemulsion templates to produce nanoparticles as drug delivery vehicles for proteins and peptides. All microemulsion and nanoparticle compounds used were pharmaceutically acceptable and biocompatible. Phase diagrams were established and characterized using polarizing light microscopy, viscosity, conductivity, electron microscopy, differential scanning calorimetry and self-diffusion NMR. An area in the phase diagrams containing optically isotropic, monophasic systems was designated as the microemulsion region and systems therein identified as solution-type microemulsions. Poly(alkylcyanoacrylate) nanoparticles prepared by interfacial polymerisation from selected microemulsions ranged from 145 to 660nm in size with a unimodal size distribution depending on the type of monomer (ethyl (2) or butyl (2) cyanoacrylate) and microemulsion template. Generally larger nanoparticles were formed by butyl (2) cyanoacrylate. Insulin was added as a model protein and did not alter the physicochemical behaviour of the microemulsions or the morphology of the nanoparticles. However, insulin-loaded nanoparticles in the CCG containing system decreased in size when using butyl (2) cyanoacrylate. This study shows that microemulsions containing sugar-based surfactants are suitable formulation templates for the formation of nanoparticles to deliver peptides. PMID- 17923348 TI - Response growth using a low-frequency suppressor. AB - Numerous psychophysical studies on two-tone suppression have been carried out. More recently, researchers have attempted to relate the magnitude of suppression to the level of suppressee. [Wojtczak, M., Viemeister, N.F., 2005. Psychophysical response growth under suppression. In: Pressnitzer, D., de Cheveigne, A., McAdams, S., Collet, L. (Eds.), Auditory Signal Processing: Physiology, Psychoaccoustics, and Models. Springer, New York, pp. 67-74] demonstrated that the magnitude of suppression for a higher-frequency, fixed-level suppressor decreases with increasing level of the suppressee. This suggests a linearization of the basilar membrane response in presence of a high-frequency suppressor. The present study expands these results to a low-frequency suppressor of varying intensity levels. Detection of a 10-ms, 4.0-kHz probe was measured under different forward-masking conditions: one with a 200-ms, 4.0-kHz masker (suppressee) presented with no suppressor and another with the same masker paired with a 2.2-kHz, 200-ms suppressor. The 4.0-kHz masker level was varied adaptively and a range of probe levels was used to measure the growth of suppression. Results indicate that (1) the magnitude of suppression increases with increasing suppressor level and (2) generally, the probe level was not related to the magnitude of suppression. PMID- 17923349 TI - Alpers syndrome with prominent white matter changes. AB - Alpers syndrome is a fatal neurogenetic disorder caused by the mutations in POLG1 gene encoding the mitochondrial DNA polymerase gamma (polgamma). Two missense variants, c.248T > C (p.L83P), c.2662G > A (p.G888S) in POLG1 were detected in a 10-year-old Chinese girl with refractory seizures, acute liver failure after exposure to valproic acid, cortical blindness, and psychomotor regression. The pathology of left occipital lobe showed neuronal loss, spongiform degeneration, astrocytosis, and demyelination. In addition, there were prominent white matter changes in a series of brain magnetic resonance imaging (MRI) and increased immunological factors in CSF. PMID- 17923350 TI - Worsening renal function and prognosis in heart failure: systematic review and meta-analysis. AB - BACKGROUND: Renal impairment is associated with increased mortality in heart failure (HF). Recently, reports suggest that worsening renal function (WRF) is another predictor of clinical outcome in HF. The present study was designed to establish the proportion of patients with HF that exhibits (WRF) and the associated risk for mortality and hospitalization by conducting a systematic review and meta-analysis. METHODS AND RESULTS: A systematic search of MEDLINE revealed 8 studies on the relationship between WRF and mortality in 18,634 patients with HF. The mortality risk associated with WRF was estimated using random-effects meta-analysis. WRF was defined as an increase in serum creatinine > or = 0.2 mg/dL or a corresponding decrease in estimated glomerular filtration rate > or = 5 mL x min x 1.73 m2. Subgroup analysis included differentiation between in- and out-hospital patients, degree of WRF and time until end point occurrence. WRF developed in 4,734 (25%) patients and was associated with a higher risk for mortality (odds ratio [OR] = 1.62; 95% confidence interval [CI] 1.45-1.82, P < .001) and hospitalization (OR = 1.30, 95% CI 1.04-1.62, P = .022). The severity of WRF was also associated with greater mortality. Patients with impaired renal function at baseline were more prone to progressive renal function loss. CONCLUSIONS: WRF predicts substantially higher rates of mortality and hospitalization in patients with HF. PMID- 17923351 TI - The effect of KW-3902, an adenosine A1 receptor antagonist, on renal function and renal plasma flow in ambulatory patients with heart failure and renal impairment. AB - BACKGROUND: The kidney is the only organ in which adenosine is a paracrine vasoconstrictor. This raises the possibility of using adenosine A1 receptor (AA1R) antagonists to selectively vasodilate the kidney in conditions, such as congestive heart failure, in which a selective decrease in renal vascular resistance would be salutary. The present study was undertaken to test the effectiveness of an AA1R antagonist as a renal vasodilator in patients with reduced kidney function superimposed on congestive heart failure. METHODS AND RESULTS: A randomized, double-blind, placebo-controlled, two-way crossover study was conducted in 32 outpatients with congestive heart failure and renal impairment (median glomerular filtration rate [GFR] 50 mL/min). Baseline GFR and renal plasma flow were assessed by iothalamate and para-amino-hippurate clearances, respectively, 3 hours before treatment. Subjects then received furosemide administered intravenously along with the AA1R antagonist, KW-3902 (rolofylline), or placebo. Clearance measurements were repeated, at intervals, throughout 8 hours beginning with the administration of the study drug. After a washout period of 3 to 8 days, subjects returned to undergo the crossover portion of the study. After the patients received KW-3902, GFR increased by 32% (P < .05 vs. placebo) and renal plasma flow increased by 48% (P < .005 vs. placebo) averaged over the ensuing 8 hours. Furthermore, those subjects who initially received KW-3902 returned for the crossover phase (median 6 days) with a persistent 10 mL/min increase in GFR more than their previous baseline (P < .05). CONCLUSIONS: AA1R activity contributes substantially to renal vascular tone in ambulatory patients with chronic congestive heart failure and impaired kidney function. Blockade of these receptors vasodilates the kidney and increases GFR. The increase in GFR seems to persist several days longer than predicted by pharmacokinetics, suggesting a resetting of one or more controllers among the complex network of physical and biological processes that interact to determine the kidney function. There may be short- or long-term benefits of using AA1R antagonists to improve kidney function in patients with congestive heart failure. PMID- 17923352 TI - A hemodynamically oriented echocardiography-based strategy in the treatment of congestive heart failure. AB - BACKGROUND: The therapeutic applicability of echocardiographic evaluations remains poorly defined in heart failure (HF). We hypothesized that an individualized echocardiography-guided strategy would be feasible and significantly reduce morbidity compared with the conventional clinically oriented treatment. METHODS AND RESULTS: We conducted a single-center clinical trial comparing an echocardiography-guided strategy aimed at achieving a near-normal hemodynamic profile and a conventional clinically oriented strategy for HF management. The echocardiography-guided strategy was based on sequential echocardiograms to evaluate hemodynamically derived parameters. Pharmacologic therapy was guided according to a predefined protocol. The primary efficacy end point was time to the first event of combined all-cause mortality and all-cause hospitalization or emergency department visit up to 1 year of follow-up. We studied 96 outpatients with HF, enrolled from 1999 to 2003, with predominantly nonischemic cause and a mean left ventricular ejection fraction of 26% +/- 6%. Event-free survival at a mean follow-up of 230 days was 58.5% with the echocardiography-guided strategy and 36.5% with the clinically based strategy (relative risk = 0.54, 95% confidence interval = 0.31-0.97, P = .04). More patients in the echocardiography-based group received high-dose loop diuretics (absolute difference of 19%, P = .02) and hydralazine (absolute difference of 30%, P < .001). Significant reductions of estimates of pulmonary artery systolic pressure (mean difference of -9 mm Hg, P = .02) and systemic vascular resistance index (mean difference of -700 dyn x sec x m2 x cm5, P = .02) were observed in the echocardiography-guided group. CONCLUSION: A hemodynamically oriented echocardiography-based strategy is feasible and decreases HF morbidity. This benefit could be attributed in part to the rational and individualized use of higher doses of diuretics and vasodilators. PMID- 17923353 TI - Hemodynamically tailored therapy in congestive heart failure: alive and well. PMID- 17923354 TI - Cardiac output and cardiopulmonary responses to exercise in heart failure: application of a new bio-reactance device. AB - BACKGROUND: Cardiopulmonary exercise testing (CPX) is widely used to evaluate heart failure (HF) patients, but lacks information about ventricular performance. There is a need for a noninvasive cardiac output (CO) measurement. METHODS AND RESULTS: Twenty-three HF patients and 13 normals underwent CPX and CO measurements with a new, noninvasive, bioreactance technology based on assessment of relative phase shifts (d phi/d t) of electric currents injected across the thorax, heart rate, and ventricular ejection time. CO and oxygen uptake (VO2) closely paralleled each another during exercise, and peak CO was strongly correlated with peak VO2 (r = 0.73, P < .001). The relationships between peak VO2 and peak cardiac index (CI) were similar for directly measured CI (r = 0.61) and noninvasive CI (r = 0.61). The CO-VO2 relationship was also similar between the current study and previous studies from the literature, as evidenced by a similar line of best fit and 95% confidence limits. Maximal CO was significantly related to indices of ventilatory efficiency, including the VE/VCO2 slope (r = -0.47, P < .01), and the oxygen uptake efficiency slope (r = 0.67, P < .01). CONCLUSION: Noninvasive measurement of CO during exercise using a novel bioreactance-based device has potentially important applications as a simple, inexpensive tool to supplement the clinical evaluation of patients with HF. PMID- 17923355 TI - Improved response to cardiac resynchronization therapy through optimization of atrioventricular and interventricular delays using acoustic cardiography: a pilot study. AB - BACKGROUND: The purpose of this pilot study was to determine the utility of acoustic cardiography for the optimization of atrioventricular (AV) and interventricular (VV) delays in cardiac resynchronization therapy (CRT). METHODS AND RESULTS: We evaluated 14 patients (86% male, mean age 64 +/- 9 years, mean time since implant 15 +/- 18 months). Subjects were enrolled >10 weeks after CRT implant. Spiroergometry and 2-dimensional/3-dimensional echocardiography were used to assess cardiac performance for "out-of-the-box" settings (baseline settings: AV 120 ms, VV 0 ms) versus optimal settings (determined by acoustic cardiography). Cardiac performance measurements were performed 6 weeks after settings were modified. Optimal AV/VV settings were determined based on the lowest electromechanical activation time (EMAT, the time from the onset of QRS to the mitral valve component of the first heart sound). Statistical analysis was performed using a paired 2-tailed Student's t-test. In comparison to "out-of-the box" settings, AV/VV delay optimization with acoustic cardiography improved cardiac performance as indicated by significant changes in work capacity, maximum oxygen uptake, oxygen pulse, ejection fraction, end-systolic volume, and velocity time integral in left ventricular outflow tract. CONCLUSIONS: AV and VV optimization by acoustic cardiography produces significant improvements in objective clinical and hemodynamic parameters in comparison to typical "out-of the-box" settings. PMID- 17923356 TI - Symptoms, depression, and quality of life in patients with heart failure. AB - BACKGROUND: Little is known about symptoms and their burden in outpatients with chronic heart failure. Diverse symptoms may be associated with poor heart failure related quality of life, and depression may be related to increased symptoms. METHODS AND RESULTS: The number of symptoms and symptom distress (physical symptoms on the Memorial Symptom Assessment Scale-Short Form), depression (Geriatric Depression Scale-Short Form), and heart failure-related quality of life (Kansas City Cardiomyopathy Questionnaire) were measured cross-sectionally in 60 patients with heart failure from two outpatient cardiology clinics. Patients experienced a mean of nine symptoms in the previous week. More than half reported shortness of breath, lack of energy, pain, feeling drowsy, or dry mouth. In unadjusted analyses, more severe depression was associated with a greater number of symptoms (r = 0.51, P < .0001) and greater overall symptom distress (r = 0.58, P < .0001). For each additional depression symptom, the number of symptoms reported increased by 0.6 after adjustment for age, race, and N-terminal pro-brain natriuretic peptide (P = .01). The number of symptoms accounted for 32% of the variance in quality of life (P < .0001). CONCLUSIONS: Patients with heart failure report a large number of distressing symptoms. Depression in patients with heart failure is associated with a greater number of symptoms, which in turn is associated with a decrease in heart failure-related quality of life. Treatment of depression and the diverse symptoms reported by patients with heart failure might significantly improve quality of life. PMID- 17923357 TI - Impact of atrial fibrillation in heart failure with normal ejection fraction: a clinical and echocardiographic study. AB - BACKGROUND: The clinical significance of atrial fibrillation (AF) in heart failure with normal ejection fraction (HFNEF) remains undetermined. METHODS AND RESULTS: We compared the clinical and echocardiographic characteristics among 238 patients hospitalized for HF. Using the cutoff of left ventricular EF of 50%, there were 146 patients with HFNEF (AF = 42) and 92 with systolic HF (AF = 30). When compared among HFNEF, the New York Heart Association (NYHA) class (2.61 +/- 0.51 versus 2.21 +/- 0.46; P < .05), 6-minute walk distance (279.7 +/- 66.0 versus 338.0 +/- 86.1 m; P < .01), quality of life score (26.1 +/- 14.3 versus 19.5 +/- 10.3; P < .05), and previous HF hospitalization were significantly worse in the AF group. These variables were significantly better in HFNEF than systolic HF with sinus rhythm, but the differences were not detected among those with AF. Patients with HFNEF and AF were associated with more severe diastolic dysfunction when compared to sinus rhythm. With a median follow-up of 10.5 months, the proportion of HFNEF patients in AF with recurrent HF hospitalization or death was significantly higher than those in sinus rhythm (28.6% versus 10.6%; P < .01). Both AF and restrictive diastolic dysfunction were independent predictors of HF hospitalization or death in HFNEF. CONCLUSION: Patients with HFNEF and AF were associated with more severe diastolic dysfunction and worse clinical outcomes than those in sinus rhythm. PMID- 17923359 TI - BNP in overweight and obese patients with heart failure: an analysis based on the BNP-LV diastolic wall stress relationship. AB - BACKGROUND: Suppressed plasma B-type natriuretic peptide (BNP) levels have been reported in obese individuals with heart failure (HF). However, the precise mechanism and significance of this relationship in the clinical setting of HF are unknown. Recently, we reported a close association between BNP levels and left ventricular end-diastolic wall stress (EDWS) as the underlying mechanism of BNP secretion; therefore, we investigated the relationship between BMI and plasma BNP levels in HF while adjusting for EDWS. METHODS AND RESULTS: In 245 consecutive patients who presented with HF, we measured plasma BNP levels and performed echocardiography and cardiac catheterization. Meridional EDWS was calculated from echocardiographic and hemodynamic parameters. The body mass index (BMI) for the total population was 22.9 +/- 0.2 kg/m2; 21% were lean (BMI < 20) and 27% overweight and obese (BMI > or = 25). The correlation between plasma BNP and EDWS [r = 0.771 (P <.001)] was more robust than any other echocardiographic or hemodynamic parameter as well as any patient's characteristics. By a univariate analysis, BMI was inversely correlated with both BNP levels and EDWS (P <.001 in both). However, a multivariable regression analysis that took into account EDWS demonstrated a significant inverse association between BMI and BNP (P <.001). CONCLUSION: The present study demonstrates that BMI determines the BNP level in a manner that is independent of the hemodynamic load in patients with HF. Overweight and obesity should be considered to be associated with the interindividual variability of plasma BNP levels in the diagnosis and management of HF patients. PMID- 17923360 TI - Utility of BNP in differentiating constrictive pericarditis from restrictive cardiomyopathy in patients with renal insufficiency. AB - BACKGROUND: Differentiating between constrictive pericarditis (CP) and restrictive cardiomyopathy (RCMP) is difficult because of similar clinical and hemodynamic presentation. Brain natriuretic peptide (BNP) has been reported a useful noninvasive biomarker to differentiate CP from RCMP; however, its utility in patients with renal insufficiency has not been evaluated. METHODS AND RESULTS: Consecutive patients with suspected CP or RCMP were enrolled. All but 7 patients underwent transseptal catheterization. BNP, renal function, and comorbid conditions were recorded at the time of the procedure. Renal function was estimated using the Cockcroft-Gault formula. Descriptive statistics, Student t test, and Mann-Whitney U test were performed; P < .05 was significant. Twenty-two patients had hemodynamically or surgically proven CP or RC. In patients with CP, 9 had at least Stage II kidney disease (GFR <90 mL/min, mean 58) and 8 had normal or Stage I kidney disease (GFR >90 mL/min, mean 118). BNP was higher in patients with CP and renal insufficiency versus those with CP and normal renal function (433 versus 116 pg/mL; P = .016). BNP in patients with CP and normal renal function was lower than in patients with RC (116 versus 728 pg/mL; P = .005). CONCLUSION: BNP has reduced clinical utility in renal insufficiency to differentiate CP from RCMP. PMID- 17923358 TI - Association of diurnal blood pressure pattern with risk of hospitalization or death in men with heart failure. AB - BACKGROUND: An altered diurnal blood pressure (BP) pattern has been linked to the risk of developing heart failure (HF). We tested whether an altered diurnal BP pattern is associated with adverse outcomes (death or hospitalization for HF exacerbation) in patients with HF. METHODS AND RESULTS: A total of 118 patients with HF were enrolled from a tertiary care HF clinic and followed for death or HF hospitalization for up to 4 years; 24-hour ambulatory BP was monitored. Forty patients (34%) had a normal BP dipping pattern (night-day ambulatory BP ratio < 0.9), 44 patients (37%) had a nondipping pattern (0.9 < or = night-day ambulatory BP ratio < 1.0), and 34 patients (29%) had a reverse dipping BP pattern (night day ambulatory BP ratio > or = 1.0). A total of 39 patients had an adverse outcome. Adverse outcome rates were the lowest in dippers and the highest in reverse dippers (log rank P = .052). Predictors of adverse outcomes, selected on the basis of log likelihood contrast, were as follows: New York Heart Association functional class (hazard ratio [HR] 1.96, 95% confidence interval [CI] 1.11 3.44), anemia (HR 2.50, 95% CI 1.23-5.08), and dipping status (HR 1.65, 95% CI 1.08-2.50). CONCLUSION: In addition to other traditional predictors, BP dipping status may be an important prognostic factor in HF. PMID- 17923361 TI - Prognostic significance and measurement of exercise-derived hemodynamic variables in patients with heart failure. AB - The peak VO2 is an important prognostic measurement in the evaluation of patients with heart failure and is used to monitor the progress of the condition, especially in selecting patients for cardiac transplantation. However, peak VO2 may be influenced by noncardiac factors such as age, sex, motivation, anemia, and muscle deconditioning. These confounding factors may diminish somewhat the prognostic power of peak VO2. Several groups have looked at exercise-derived variables beyond peak VO2 to assess whether a more direct assessment of cardiac function, using exercise-derived hemodynamic variables, may yield more precise prognostic information than standard cardiopulmonary-derived data. This article reviews the evidence that cardiac work related to exercise may enhance the prognostic value of peak VO2 in the evaluation of patients with heart failure and briefly discusses the available methods for measuring these parameters. PMID- 17923362 TI - Increased myosin heavy chain-beta with atrial expression of ventricular light chain-2 in canine cardiomyopathy. AB - BACKGROUND: Dilated cardiomyopathy is a naturally occurring disease in humans and dogs. Human studies have shown increased levels of myosin heavy chain (MHC)-beta in failing ventricles and the left atria (LA) and of ventricular light chain (VLC)-2 in the right atria in dilated cardiomyopathy. METHODS AND RESULTS: This study evaluates the levels of MHC-beta in all heart chambers in prolonged canine right ventricular pacing. In addition, we determined whether levels of VLC2 were altered in these hearts. Failing hearts demonstrated significantly increased levels of MHC-beta in the right atria, right atrial appendage, LA, left atrial appendage (LAA), and right ventricle compared with controls. Significant levels of VLC2 were detected in the right atria of paced hearts. Differences in MHC-beta expression were observed between the LA and the LAA of paced and control dogs. MHC-beta expression was significantly greater in the LA of paced and control dogs compared with their respective LAA. CONCLUSIONS: The cardiac myosin isoform shifts in this study were similar to those observed in end-stage human heart failure and more severe than those reported in less prolonged pacing models, supporting the use of this model for further study of end-stage human heart failure. The observation of consistent differences between sampling sites, especially LA versus LAA, indicates the need for rigorous sampling consistency in future studies. PMID- 17923363 TI - Reduced delayed rectifier K+ current, altered electrophysiology, and increased ventricular vulnerability in MLP-deficient mice. AB - BACKGROUND: Mice with a knockout (KO) of muscle LIM protein (MLP) exhibit many morphologic and clinical features of human cardiomyopathy. In humans, MLP expression is downregulated both in ischemic and dilative cardiomyopathy. In this study, we investigated the effects of MLP on the electrophysiologic phenotype in vivo and on outward potassium currents. METHODS AND RESULTS: MLP-deficient (MLPKO) and wild-type (MLPWT) mice were subjected to long-term electrocardiogram (ECG) recording and in vivo electrophysiologic study. The whole-cell, patch-clamp technique was applied to measure voltage dependent outward K+ currents in isolated cardiomyocytes. Long-term ECG revealed a significant prolongation of RR mean (108 +/- 9 versus 99 +/- 5 ms), P (16 +/- 3 versus 14 +/- 1 ms), QRS (17 +/- 3 versus 13 +/- 1 ms), QT (68 +/- 8 versus 46 +/- 7 ms), QTc (66 +/- 6 versus 46 +/- 7 ms), JT (51 +/- 7 versus 34 +/- 7 ms), and JTc (49 +/- 5 versus 33 +/- 7 ms) in MLPKO versus MLPWT mice (P < .05). During EP study, QT (80 +/- 8 versus 58 +/- 7 ms), QTc (61 +/- 6 versus 45 +/- 5 ms), JT (62 +/- 9 versus 43 +/- 6 ms), and JTc (47 +/- 5 versus 34 +/- 5 ms) were also significantly prolonged in MLPKO mice (P < .05). Nonsustained VT was inducible in 9/16 MLPKO versus 2/15 MLPWT mice (P < .05). Analysis of outward K+ currents in revealed a significantly reduced density of the slowly inactivating outward K+ current IK, slow in MLPKO mice (11 +/- 5 pA/pF versus 18 +/- 7 pA/pF; P < .05). CONCLUSION: Mice with KO of MLP exhibit significant prolongation of atrial and ventricular conduction and an increased ventricular vulnerability. A reduction in repolarizing outward K+ currents may be responsible for these alterations. PMID- 17923364 TI - Prognostic value of pulse pressure in heart failure. PMID- 17923365 TI - Pulmonary malignancy in silicosis: factors associated with radiographic detection. AB - The aim was to assess radiographic features of pulmonary malignancies in silicosis and to reveal confounding factors in their detection. A total of 501 silicosis patients were followed up between 1982 and 2003. Sixty pulmonary malignancies were pathologically confirmed in 54 (10.8%) patients. Two radiologists reviewed serial radiographs of these patients to determine radiographic features of tumor (size, margin, nodule or consolidation, localization, overlying structures) and silicosis (profusion of pneumoconiotic opacities, progressive massive fibrosis (PMF), hilar lymphadenopathy). Eleven tumors were radiographically negative. Forty-nine tumors were retrospectively visible with radiograph. Of these, 15 tumors were clinically detected with radiograph, but 25 were missed. The remaining nine tumors became radiographically positive after positive sputum cytology. There were no differences between missed and detected nodules in terms of radiographic findings. The mean tumor size was 30mm (range: 15-90mm) and was significantly larger in patients with PMF or hilar lymphadenopathy than in those without (35mm vs. 24mm, p=0.006; 33mm vs. 24mm, p=0.038, respectively). This was correlated with background profusion of small pneumoconiotic opacities (r=0.433, p=0.024). Retrospective reading tests by three radiologists showed correct localization of tumor in 75%, however, the correct diagnosis with a high confidence was reached in only 54%. In conclusion, radiographic detection of malignancy in silicosis proved a difficult task and no single radiographic finding was found to be associated with missing the tumor. The presence of PMF, hilar lymphadenopathy and profusion of small pneumoconiotic nodules affected tumor size at detection. PMID- 17923366 TI - Sonographic assessment of splanchnic arteries and the bowel wall. AB - The intestinal wall can be visualized using high resolution transabdominal ultrasound. The normal intestinal wall thickness in the terminal ileum, cecum, and right and left colon is <2mm when examined with graded compression. It is important to appreciate that a contracted intestinal segment can be misinterpreted as a thickened wall. Vascularisation can be mainly displayed in the second hyperechoic layer (submucosal layer) as well as vessels penetrating the muscularis propria. Imaging of the gastrointestinal wall is dependent on the experience of the examiner as well dependent on the equipment used. Acute or chronic inflammation of the intestinal wall is accompanied by increased perfusion of the mesentery, which can be displayed non-quantitatively with colour duplex. In contrast, ischemia is characterised by hypoperfusion of the mesenteric arteries and the bowel wall. The most promising sonographic approach in assessing splanchnic arteries and the bowel wall is combining the analysis of superior and inferior mesenteric inflow by pulsed Doppler scanning (systolic and diastolic velocities, resistance index) with the end-organ vascularity by colour Doppler imaging diminishing the influence of examination technique only displaying bowel wall vascularity. Colour Doppler imaging has been described as helpful in a variety of gastrointestinal disorders, particularly in patients with Crohn's disease, celiac disease, mesenteric artery stenosis and other ischemic gastrointestinal diseases, graft versus host disease and hemorrhagic segmental colitis. PMID- 17923367 TI - A non-radioactive method for detecting neutralizing antibodies against therapeutic proteins in serum. AB - The presence of neutralizing antibodies against protein therapeutics continues to cause concern in the biomedical field. These antibodies not only reduce the efficacy of the protein therapeutics, but may also block the normal function of their endogenous counterparts, which can result in serious health risks to the patient. To date, a limited number of in vitro cell-based bioassays for detecting neutralizing antibodies against therapeutic proteins have been developed. However, many of the existing assays involve the use of radioactive materials. We have established a novel and non-radioactive bioassay system for detecting neutralizing antibodies in patient serum samples. Our assay measures the cell metabolic activities that are closely associated with cell proliferation and apoptosis. The biologic effect of the therapeutic protein and the capability of the antibodies to neutralize the therapeutics are reflected by changes of the cellular metabolic activities triggered by the administration of the therapeutics or presence of the anti-therapeutic protein antibodies. Compared with existing assays, this new assay is equally or more sensitive, and completely eliminates the use of radioactive materials. PMID- 17923368 TI - Impact of indolent inflammation on neonatal hypoxic-ischemic brain injury in mice. AB - This report describes a new experimental model to evaluate the effect of a recurrent systemic inflammatory challenge, after cerebral hypoxia-ischemia in immature mice, on the progression of brain injury. Treatment with a low dose of lipopolysaccharide (E. coli O55:B5, 0.2mg/kg for 3 days, then 0.1mg/kg for 2 days) daily for 5 days after unilateral cerebral hypoxia-ischemia (right carotid ligation followed by 35min in 10% O2) in 10-day-old mice resulted in increased right forebrain tissue damage (35.6% reduction in right hemisphere volume compared to 20.6% reduction in saline-injected controls), in bilateral reductions in corpus callosum area (by 12%) and myelin basic protein immunostaining (by 19%), and in suppression of injury-related right subventricular zone cellular proliferation. The post-hypoxic-ischemic lipopolysaccharide regimen that amplified brain injury was not associated with increased mortality, nor with changes in body temperature, weight gain or blood glucose concentrations. The results of the present study demonstrate that systemic inflammation influences the evolution of tissue injury after neonatal cerebral hypoxia-ischemia and may also impair potential recovery mechanisms. PMID- 17923370 TI - The NADPH quinone reductase MdaB confers oxidative stress resistance to Helicobacter hepaticus. AB - An mdaB mutant strain in a quinone reductase (MdaB) of Helicobacter hepaticus type strain ATCC51449 was constructed by insertional mutagenesis, and the MdaB protein was purified and compared to the Helicobacter pylori enzyme. While wild type H. hepaticus cells could tolerate 6% O(2) for growth, the mdaB strain was clearly inhibited at this oxygen level. Disruption of the gene downstream of mdaB (HH1473) did not affect the oxidative stress phenotype of the strain. The mdaB mutant was also more sensitive to oxidative stress reagents such as H(2)O(2), cumene hydroperoxide, t-butyl hydroperoxide, and paraquat. All H. hepaticus mdaB strains isolated constitutively up-expressed another oxidative stress-combating enzyme, superoxide dismutase; this is in contrast to H. pylori mdaB strains. H. hepaticus MdaB is a flavoprotein catalyzing quinone reduction using a two electron transfer mechanism from NAD(P)H to quinone. The H. hepaticus enzyme specific activity was far less than for the H. pylori enzyme purified in the same manner. PMID- 17923372 TI - Evaluation of genetic variability of sorghum (Sorghum bicolor L. Moench) in northwestern Morocco by ISSR and RAPD markers. AB - The study of the genetic variability of the Moroccan landraces of sorghum constitutes a necessary step that can be exploited in the programs of improvement and valorisation of this marginalized species. The aim of this investigation is to evaluate the variability of sorghum populations and to establish their phylogenetic relations using RAPD and ISSR markers. Sampling was taken in 33 fields of northern regions where this species is most cultivated. Individual plants (398) were collected in 13, 11, 5, and 4 fields of Larache, Tangier, Chefchaouen, and Tetouan, respectively. Thirty-eight RAPD primers and four ISSR primers were used. The percentage of polymorphic fragments revealed with ISSR (98%) is higher than the one revealed with RAPD (85%). The level of the variability obtained through the two techniques is very high. Nevertheless, ISSR markers revealed more diversity than RAPD (0.995+/-0.006 against 0.946+/-0.031). The classification based on Jaccard's similarity index distinguished the totality of fields. Data analysis revealed a genetic structure that is closely related to the micro-geographical repartition of the different fields. PMID- 17923371 TI - Interactive effects of salinity and iron deficiency in Medicago ciliaris. AB - In calcareous salt-affected soils, iron availability to plants is subjected to the effects of both sodium and bicarbonate ions. Our aim was to study interactive effects of salinity and iron deficiency on iron acquisition and root acidification induced by iron deficiency in Medicago ciliaris L., a species commonly found in saline ecosystems. Four treatments were used: C, control treatment, complete medium (CM) containing 30 microM Fe; S, salt treatment, CM with 75 mM NaCl; D, deficient treatment, CM containing only 1 microM Fe; DS, interactive treatment, CM containing 1 microM Fe with 75 mM NaCl. Our study showed that plant growth and chlorophyll content were much more affected by the interactive treatment than by iron deficiency or by the salt treatment, indicating an additive effect of these constraints in DS plants. These results could be partially explained by Na accumulation in shoots as well as a limitation of nutrient uptake such as Fe and K under salt stress, under iron deficiency, and especially under their combined effect. The study also showed that root acidification was deeply diminished when iron deficiency was associated with salinity. This probably explained the decrease of Fe uptake and suggested that root proton pump activity would be inhibited by salinity. PMID- 17923369 TI - Chronic cigarette smoking and heavy drinking in human immunodeficiency virus: consequences for neurocognition and brain morphology. AB - Alcohol use disorders (AUD) and chronic cigarette smoking are common among individuals with human immunodeficiency virus infection (HIV). Concurrent AUD in HIV is related to greater abnormalities in brain morphology and neurocognition than either condition alone. However, the potential influence of chronic smoking on brain morphology and neurocognition in those concurrently afflicted with AUD and HIV has not been examined. The goal of this retrospective analysis was to determine if chronic smoking affected neurocognition and brain morphology in a subsample of HIV-positive non-treatment-seeking heavy drinking participants (HD+) from our earlier work. Regional volumetric and neurocognitive comparisons were made among age-equivalent smoking HD+(n=17), nonsmoking HD+ (n=27), and nonsmoking HIV-negative light drinking controls (n=27) obtained from our original larger sample. Comprehensive neuropsychological assessment evaluated multiple neurocognitive domains of functioning and for potential psychiatric comorbidities. Quantitative volumetric measures of neocortical gray matter (GM), white matter (WM), subcortical structures, and sulcal and ventricular cerebral spinal fluid (CSF) were derived from high-resolution magnetic resonance images. The main findings were (1) smoking HD+ performed significantly worse than nonsmoking HD+ on measures of auditory-verbal (AV) learning, AV memory, and cognitive efficiency; (2) relative to controls, smoking HD+ demonstrated significantly lower neocortical GM volumes in all lobes except the occipital lobe, while nonsmoking HD+ showed only lower frontal GM volume compared with controls; (3) in the HD+ group, regional brain volumes and neurocognition were not influenced by viremia, highly active antiretroviral treatment, or Center for Disease Control symptom status, and no interactions were apparent with these variables or smoking status. Overall, the findings suggested that the direct and/or indirect effects of chronic cigarette smoking created an additional burden on the integrity of brain neurobiology and neurocognition in this cohort of HIV positive heavy drinkers. PMID- 17923374 TI - NaCl as a physiological modulator of proline metabolism and antioxidant potential in Phyllanthus amarus. AB - Some medicinal plants need to be cultivated commercially in order to meet the ever-increasing demand for medicinal plants for the indigenous systems of medicine as well as for the pharmaceutical industry; in this regard, it seems significant to test the important medicinal plants for their salt-tolerance capacity, with a view to exploiting the saline lands for medicinal plant cultivation. Phyllanthus amarus plants were grown in the presence of NaCl in order to study the effect of NaCl (80 mM NaCl) in the induction of oxidative stress in terms of lipid peroxidation (TBARS content), H2O2 content, osmolyte concentration, proline(PRO)-metabolizing enzymes, and antioxidant enzyme activities. Groundwater was used for irrigation of control plants. Plants were uprooted randomly on 90 days after sowing (DAS). NaCl-stressed plants showed increased TBARS, H2O2, glycine betaine (GB), and PRO contents, whereas NaCl uptake decreased proline oxidase (PROX) activity and increased gamma-glutamyl kinase (gamma-GK) activity when compared to control. The antioxidant enzymes superoxide dismutase (SOD), peroxidase (POX) and catalase (CAT) were increased under salinity. PMID- 17923373 TI - Responses of antioxidant potentials in Dioscorea rotundata Poir. following paclobutrazol drenching. AB - The effect of paclobutrazol (PBZ) treatments on the antioxidant metabolism of white yam (Dioscorea rotundata Poir.) was investigated in the present study. PBZ @ 15 mg l(-1) plant(-1) was given to plants by soil drenching, 30, 60, and 90 days after planting (DAP). The non-enzymatic antioxidant contents like ascorbic acid (AA), reduced glutathione (GSH) and alpha-tocopherol (alpha-toc), activities of antioxidant enzymes like superoxide dismutase (SOD), ascorbate peroxidase (APX), polyphenol oxidase (PPO) and catalase (CAT) were extracted and assayed on 100 DAP from leaf, stem and tubers of both control and PBZ treated plants. It was found that PBZ has a profound effect on the antioxidant metabolism and caused an enhancement in both non-enzymatic and enzymatic antioxidant potentials under treatments in white yam. Our results have good significance, as this increase the innate antioxidant potential of this food crop, which is helpful to satisfy the needs of antioxidants in diet and thereby make it an economically important food crop. PMID- 17923375 TI - Changes in antioxidant potential and sink-organ dry matter with pigment accumulation induced by hexaconazole in Plectranthus forskholii Briq. AB - An investigation was conducted in Plectranthus forskholii by giving it different concentrations (10, 15, 20, 25, and 30 mg L(-1)) of hexaconazole, a fungicide cum plant-growth regulator, in order to find out its effects on growth, pigment composition, and antioxidant potential. The treatments were given as soil drenching on different growth stages. All the concentrations of hexaconazole used significantly decreased the stem length and leaf area, whereas side branches, fresh and dry matter content, photosynthetic pigments, and antioxidant potential were increased. The number of tubers, length and girth of tubers, fresh and dry weight of tubers and tuber pigments were found to be the highest at a 25 mg L(-1) concentration of hexaconazole. Hexaconazole application at 25 mg L(-1) concentration was found to be more effective than 10, 15, 20, 25, and 30 mg L(-1) in promoting fresh and dry weight of root tuber over 165 days after planting. The pigments like chlorophyll, carotenoid; anthocyanins, xanthophylls and antioxidants such as ascorbic acid, reduced glutathione and total phenol were significantly increased under hexaconazole treatment when compared to untreated control plants. PMID- 17923376 TI - Multiple-contrast X-ray micro-CT visualization of colon malformations and tumours in situ in living mice. AB - The development of new therapeutic approaches against colorectal cancer requires preclinical studies in mice. In vivo imaging could greatly facilitate these trials, but the small size of the animals is a major limitation for the direct visualization of intestinal tissue. Here we report a method of in vivo imaging of the mouse intestine based on X-ray micro-computed tomography using multiple contrast agents. This method was validated in the model of non-cancerous polyp like heteroplasia that spontaneously develops in the caecum area of Cdx2+/- mutant mice and in the model of colon adenocarcinoma induced by administration of the chemical carcinogen azoxymethane. As a simple and non-invasive method, multiple-contrast X-ray micro-computed tomography is appropriate for pre-clinical studies of intestinal diseases in living mice. PMID- 17923378 TI - Evaluation of the social bond: a new method tested in Mus spicilegus. AB - Innovative and fruitful studies of social bonds have been developed in recent years, although the methods used to establish the existence of a social bond between two individuals have not evolved significantly. Two types of paradigms have been currently used: the separation-reunion paradigm, which evaluates the distress caused by the disruption of the social bond, and choice paradigms, which test the specificity of the bond to a given individual. We have developed a new paradigm based on the idea that the cost an individual was ready to pay in order to gain access to a conspecific depended on the strength of the social bond between the two individuals. To test our paradigm we used mound-building mice, Mus spicilegus that present, in both males and females, a level of tolerance that differs greatly according to the degree of familiarity between the individuals. Our new method for testing social bond revealed unsuspected differences between males and females. Our results suggested that, at least in Mus spicilegus, strong social bonds were not necessary to the development of a high level of tolerance between individuals. PMID- 17923377 TI - Pair bonding in the wild mouse Mus spretus: inference on the mating system. AB - The mating system of murids remains largely unknown, with controversial reports. Reevaluation of previous data reveals monogamous features in the Algerian mouse from the South of France, Mus spretus. To test this hypothesis, we performed dyadic encounters toward three kinds of opponents from the other sex (his/her own mate, a mouse from an unknown pair, and an isolated individual), and compared the results to those of a related acknowledgedly polygynous related species, the house mouse M. musculus domesticus. We expected more affiliative behaviours toward its mate from the M. spretus male, and a higher interest toward unknown females in the house mouse. Females should not exhibit striking differences toward their partner, contrary to their behaviour toward unknown males. Results showed the predicted behaviour for males as well as for females: more amicable, but also sexual, toward its mate for M. spretus male, the unknown protagonists eliciting the greatest social interest in the case of the house mouse. PMID- 17923380 TI - Dopaminergic neurons are preferentially sensitive to long-term rotenone toxicity in primary cell culture. AB - Parkinson's disease (PD) is a chronic neurodegenerative disorder characterized by the death of dopaminergic neurons in the substantia nigra pars compacta (SNpc) and the subsequent decrease of dopamine levels in the striatum. Epidemiological studies indicate environmental pollutants as a causative factor of sporadic PD. Experimental cell culture models have the inherent problem to mimic long-lasting neurodegeneration and to tackle its time-concentration relationship. The present study was designed to investigate the sensitivity of primary dopaminergic neurons to long-term rotenone exposure relevant to PD. Primary cultures prepared from embryonic mouse mesencephala were treated with nanomolar concentrations of rotenone (1, 3, 5, 10nM) on the 6th day in vitro (DIV) for 2, 4 and 6 days. The number of tyrosine hydroxylase immunoreactive (TH(+)) neurons and total hematoxylin-stained nuclei were counted. Astrocyte density was qualitatively evaluated by anti-glial fibrillary acidic protein (anti-GFAP) immunocytochemistry. It was found that dopaminergic neurons were highly sensitive to long-term rotenone treatment. Rotenone in a concentration- and time-dependent manner decreased the number of TH(+) neurons and led to degenerative changes of their morphology. Counting of the total cell number revealed a significant deleterious effect on the overall culture after 6 days of rotenone exposure. However, our study demonstrates a higher sensitivity of dopaminergic neurons to long-term exposure to nanomolar concentrations of rotenone. Other cells in the culture including non-dopaminergic neurons and glia cells appeared less affected compared to dopaminergic neurons. PMID- 17923379 TI - Sensitivity of different cell lines to phototoxic effect of disulfonated chloroaluminium phthalocyanine. AB - Photodynamic therapy (PDT) is a treatment for cancer involving three key components: sensitizer, light and tissue oxygen. A sensitizer is a chemical compound that can be excited by light of a specific wavelength. Phthalocyanine ClAlPcS(2), belonging among the promising second generation of sensitizers, was evaluated as an inducer of photodamage on NIH3T3 (mouse fibroblasts), B16 (mouse melanoma), MCF7 (human breast adenocarcinoma) and G361 (human melanoma) cell lines. A semiconductor laser was used as a source for evocation of the photodynamic effect. We report the influence of various concentrations of the sensitizer in combination with laser irradiation on the photodamage of cells. Viability of cells was determined by means of molecular probes (Calcein AM and ethidium homodimer) for fluorescence microscopy. The quantitative changes of cell viability in relation to sensitizer concentrations and laser irradiation were proved by fluorometric measurement. We detected phototoxicity evoked by laser irradiated sensitizer in all studied cell lines. In addition, the viability studies showed that G361 melanoma cells and MCF7 breast adenocarcinoma cells were more sensitive than NIH3T3 mouse fibroblasts and B16 mouse melanoma to photodynamic damage induced by ClAlPcS(2). PMID- 17923381 TI - Screening for social phobia in medical in- and outpatients with the German version of the Social Phobia Inventory (SPIN). AB - OBJECTIVE: To evaluate the German version of the Social Phobia Inventory (SPIN) as a screening device and to report corresponding cut-off scores for different populations. METHOD: In Study 1, 2043 subjects from a representative sample completed the SPIN. Cut-off values were established on the basis of means and standard deviations. In Study 2, different aspects of validity were examined in a clinical sample comprising 164 subjects, including social phobic individuals, individuals with other anxiety disorders and depression, and non-clinical control subjects. Internal consistency was evaluated. Convergent and divergent validity were explored using several established measures. Finally, the sensitivity and specificity of the German SPIN with regard to social anxiety classification were investigated by means of receiver operating characteristics (ROC) analyses. RESULTS: In Study 1, mean scores and standard deviations were used to determine cut-off scores for the German SPIN. In Study 2, excellent internal consistency and good convergent and divergent validity were obtained. ROC analyses revealed that the German SPIN performed well in discriminating between social phobic individuals on the one hand and psychiatric and non-psychiatric controls on the other. A cut-off score of 25 represented the best balance between sensitivity and specificity. CONCLUSION: Comparable to the original version, the German SPIN demonstrates solid psychometric properties and shows promise as an economic, reliable, and valid screening device. PMID- 17923382 TI - Development of a novel vascular simulator and injury model to evaluate smooth muscle cell response following balloon angioplasty. AB - Following balloon angioplasty, denudation of endothelial cells exposes vascular smooth muscle cells (SMCs) to normally unseen shear forces from blood flow. In vivo studies investigate the response to angioplasty injury, but limited studies have been performed using in vitro systems. In order to study SMC response in vitro, a concurrent shear and tensile forces simulator has been developed to provide clinically significant levels of strain and shear stresses in addition to simulating forces similar to those during balloon angioplasty. In this acute study (8 hr), rat aortic SMCs demonstrated significant cell proliferation following applied increased tensile forces of angioplasty injury and shear exposure when compared to lower levels of tensile exposure similar to a normal physiological level, with an average 75% increase in the number of cells of the injury group compared to the normal dynamic group. SMCs exposed to balloon angioplasty injury and concurrent shear and tensile mechanical forces demonstrated decreased expression of the contractile phenotypic marker smooth muscle alpha-actin. These findings demonstrate the efficacy of the developed model for in vitro angioplasty and the simulated mechanical environment to the cells. This provides an in vitro model to isolate the effects of concurrent mechanical forces and could also potentially act as a preliminary step toward use in pharmaceutical research for reduction or prevention of SMC proliferation due to altered mechanical forces during endovascular procedures. PMID- 17923384 TI - Open repair of aortic aneurysms involving the renal vessels. AB - This retrospective review examines the open surgical repair of intact juxtarenal (JRAAs) and suprarenal (SRAAs) aortic aneurysms to estimate effects on survival and renal function. Patients undergoing open repair of JRAA and SRAA were identified. Preoperative medical comorbidities and perioperative and late outcomes were recorded. Primary end points were survival (perioperative and long term survival) and changes in renal function (acute tubular necrosis [ATN], acute dialysis, and late functional decline). Associations between outcomes and clinical variables were examined using univariate and multivariate techniques. Between December 1996 and September 2006, 678 patients underwent open repair of aortic aneurysms, including 150 aneurysms involving the renal vessels (134 JRAAs, 16 SRAAs). Perioperative mortality was 3% and long-term survival was 69% at 5 years. Fourteen percent of patients experienced ATN, and 7% required acute in hospital dialysis. Late renal function remained unchanged or improved in 75%. These results demonstrate a perioperative mortality and renal complication rate in keeping with previous reports of open abdominal aortic aneurysm repairs involving the renal vessels. Future implementation of branch and fenestrated aortic endografts to treat similar aneurysms should approximate these results prior to widespread acceptance. PMID- 17923383 TI - Redo pedal bypass surgery after pedal graft failure: gain or gadget? AB - Pedal bypass failure is not always associated with limb loss. Management of critical limb ischemia after failure is controversial. The aim of this study is to evaluate the results of redo bypass procedures to foot arteries in the absence of alternative tibial outflow arteries. Data of patients undergoing redo pedal bypass within a 14-year period were reviewed. The outcome after redo pedal bypass in patients whose original pedal bypass failed within 30 days versus those in patients whose original pedal bypass failed more than 30 days after the original pedal bypass were reviewed. Society for Vascular Surgery reporting standards were applied. Out of 335 pedal bypass grafts, 22 (6.6%) pedal redo bypass procedures were identified in 20 patients performed after previous pedal graft failure: 64% were male, mean age 67.7 +/- 9.5 years, diabetes 90.9%, hypertension 90.9%, coronary disease 68.2%, renal disease 18.2%. Seven patients were operated for early failure and 15 for late failure (median 193 days). The graft conduit at the first operation was ipsilateral greater saphenous vein (GSV) in 18 (81.8%), alternative vein in three (13.6%), and one expanded polytetrafluoroethylene. Redo graft conduits were as follows: ipsilateral GSV in nine (40.9%), arm vein in six (27.3%), contralateral GSV in two (9.1%), "other veins" in two (9.1%), and homologous artery in three (13.6%). The same target artery was used in 81.8%, at the initial site in 54.5% and more distally in 27.3%. Redo revascularization for early failure was successful only once. Median follow-up after late redo was 23.7 months. Seven redo grafts performed after late pedal graft failure failed after a median of 115 days. The availability of adequate autologous conduit is the limiting factor for redo procedures. Lack of alternative outflow sites adds to the difficulty of target artery dissection. Redo pedal bypass surgery after early pedal bypass failure is associated with very poor patency and limb salvage. Acceptable patency and extension of limb salvage can be achieved with redo procedures for late pedal bypass failure. PMID- 17923385 TI - Management of effort thrombosis of the subclavian vein: today's treatment. AB - "Spontaneous" subclavian vein ("effort") thrombosis is usually related to extrinsic compression of the vein at the costoclavicular junction. Our experience with this entity over the past decade was reviewed with specific focus on the role of thrombolysis and our selection algorithm. All patients treated for this problem at the University of Rochester over the past decade were identified and records retrospectively reviewed. From 1996 to June 2006, a total of 34 patients with spontaneous complete occlusion of the subclavian vein documented by venography were treated, half with a history of an antecedent exertional factor. Catheter-directed thrombolysis prior to planned immediate thoracic outlet decompression (TOD) was performed in 26 patients, while TOD alone was performed in eight. Time since onset of symptoms was the major factor influencing the decision, being a mean of 5.5 days in the 26 referred for lysis but 1 month to many years in the group who underwent surgery alone. In patients undergoing lysis, flow was restored in 16 (62%), of whom nine had a residual lesion. All but two of the 26 who received thrombolysis then underwent TOD with or without angioplasty, and 13 underwent venous reconstruction as well (eight of the 16 in whom patency had been restored and five of eight in whom it had not). Patients not undergoing lysis were managed by TOD, with five (62%) undergoing decompression alone and three (38%) undergoing formal venous reconstruction. Thrombolysis was not attempted (eight) or unsuccessful (four) in all 12 patients whose symptoms had been present for more than 14 days at presentation. At mean follow-up of 33 months, symptom resolution was almost universal. Primary patency at 5 years was 84% in the thrombolysis group and 83% in the TOD-only group. In conclusion, the primary factor influencing treatment choice at our institution has been time since onset of symptoms. Patients presenting soon after symptom onset underwent thrombolysis followed by TOD, while patients presenting with chronic symptoms underwent TOD alone; in both cases venous reconstruction was based on residual findings after lysis. We have not had successful thrombolysis in a patient presenting with 14 days or more of symptoms. While optimal therapy cannot be defined based on this retrospective review, our algorithm resulted in excellent overall long-term patency and symptom relief. PMID- 17923386 TI - Increased risk of cancer in the offspring of female electronics workers. AB - There is limited evidence on the hypothesis that maternal occupational exposure near conception increases the risk of cancer in offspring. This study is to investigate whether women employed in an electronics factory increases childhood cancer among first live born singletons. We linked the databases of Birth Registration and Labor Insurance, and National Cancer Registry, which identified 40,647 female workers ever employed in this factory who gave 40,647 first live born singletons, and 47 of them developed cancers during 1979-2001. Mothers employed in this factory during their periconceptional periods (3 months before and after conception) were considered as exposed and compared with those not employed during the same periods. Poisson regression model was constructed to adjust for potential confounding by maternal age, education, sex, and year of birth. Based on 11 exposed cases, the rate ratio of all malignant neoplasms was increased to 2.26 [95% confidence interval (CI), 1.12-4.54] among children whose mothers worked in this factory during periconceptional periods. The RRs were associated with 6 years or less (RR=3.05; 95% CI, 1.20-7.74) and 7-9 years (RR=2.49; 95% CI, 1.26-4.94) of education compared with 10 years or more. An increased association was also found between childhood leukemia and exposed pregnancies (RR=3.83; 95% CI, 1.17-12.55). Our study suggests that maternal occupation with potential exposure to organic solvents during periconception might increase risks of childhood cancers, especially for leukemia. PMID- 17923387 TI - Nuclear organization and morphology of serotonergic neurons in the brain of the Nile crocodile, Crocodylus niloticus. AB - The present study describes the location and nuclear organization of the serotonergic system in a representative of the order Crocodylia, the Nile crocodile (Crocodylus niloticus). We found evidence for serotonergic neurons in three regions of the brain, including the diencephalon, rostral and caudal brainstem, as previously reported in several other species of reptile. Within the diencephalon we found neurons in the periventricular organ of the hypothalamus, but not in the infundibular recess as noted in some other reptilian species. In addition we found serotonergic neurons in the pretectal nucleus, this being the first description of these neurons in any species. Within the rostral brainstem we found medial and lateral divisions of the superior raphe nucleus and a widely dispersed group of neurons in the tegmentum, the superior reticular nucleus. In the caudal brainstem we observed the inferior raphe nucleus and the inferior reticular nucleus. While much of the serotonergic system of the Nile crocodile is similar to that seen in other reptiles the entire suite of features appears to distinguish the crocodile studied from the members of the Squamate (lizards and snakes) and Testudine (turtles, tortoises and terrapins) reptiles previously studied. The observations are suggestive of order-specific patterns of nuclear organization of this system in the reptiles, reflecting potential evolutionary constraints in the mutability of the nuclear organization as seen for similar systems in mammals. PMID- 17923388 TI - Cognitive, behavior and intervention outcome in young children with autism. AB - The relations between cognition and autism severity, head size and intervention outcome, were examined. Change in cognitive level with intervention was measured in children with autism and compared to children with developmental disabilities (DD). Eighty-one children (mean age 25.9 months) with autism (n=44) and DD (n=37) were assessed at pre- and post 1 year of intervention. Cognitive abilities and autism severity were measured by standardized tests. Three pre-intervention cognitive level groups: normal (IQ>90), borderline (70